Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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TITLE OF THE INVENTION
ALPHA 1a ADRENERGIC RECEPTOR ANTAGONISTS
This application claims the benefit of U.S. Provisional
Application No. 60/049,921, filed June 18, 1997.
FIELD OF THE INVENTION:
This invention relates to certain novel compounds and
derivatives thereof, their synthesis, and their use as alpha 1a
adrenoceptor antagonists. More particularly, the compounds of the
present invention are useful for treating benign prostatic hyperplasia
(BPH).
BACKGROUND OF THE INVENTION
Human adrenergic receptors are integral membrane
proteins which have been classified into two broad classes, the alpha and
the beta adrenergic receptors. Both types mediate the action of the
peripheral sympathetic nervous system upon binding of catecholamines,
norepinephrine and epinephrine.
Norepinephrine is produced by adrenergic nerve endings,
while epinephrine is produced by the adrenal medulla. The binding
affinity of adrenergic receptors for these compounds forms one basis of
the classification: alpha receptors bind norepinephrine more strongly
than epinephrine and much more strongly than the synthetic compound
isoproterenol. The binding affinity of these hormones is reversed for the
beta receptors. In many tissues, the functional responses, such as
smooth muscle contraction, induced by alpha receptor activation are
opposed to responses induced by beta receptor binding.
Subsequently, the functional distinction between alpha and
beta receptors was further highlighted and refined by the
pharmacological characterization of these receptors from various
animal and tissue sources. As a result, alpha and beta adrenergic
receptors were further subdivided into alpha 1~ alpha 2~ 131, and 132
subtypes. Functional differences between alpha 1 and alpha 2 receptors
have been recognized, and compounds which exhibit selective binding
between these two subtypes have been developed.
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For a general background on the alpha adrenergic
receptors, the reader's attention is directed to Robert R. Ruffolo, Jr., c~c-
Adrenorecentors~ Molecular Biolo~v Biochemistry and Pharmacolo~v,
(Proeress in Basic and Clinical Pharmacolo~v series, Karger, 1991),
wherein the basis of alpha 1/alpha 2 subclassification, the molecular
biology, signal transduction (G-protein interaction and location of the
significant site for this and ligand binding activity away from the 3'-
terminus of alpha adrenergic receptors), agonist structure-activity
relationships, receptor functions, and therapeutic applications for
compounds exhibiting alpha-adrenergic receptor affinity was explored.
The cloning, sequencing and expression of alpha receptor
subtypes from animal tissues has led to the subclassification of the
alpha 1 receptors into alpha Id (formerly known as alpha la or Ialld),
alpha 1b and alpha la (formerly known as alpha lc) subtypes. Each
alpha 1 receptor subtype exhibits its own pharmacologic and tissue
specificities. The designation "alpha la" is the appellation recently
approved by the IUPHAR Nomenclature Committee for the previously
designated "alpha lc" cloned subtype as outlined in the 1995 Receptor
and Ion Channel Nomenclature Supplement {Watson and Girdlestone,
1995). The designation alpha la is used throughout this application to
refer to this subtype. At the same time, the receptor formerly designated
alpha la was renamed alpha 1d. The new nomenclature is used
throughout this application. Stable cell lines expressing these alpha 1
receptor subtypes are referred to herein; however, these cell lines were
deposited with the American Type Culture Collection (ATCC) under the
old nomenclature. For a review of the classification of alpha 1
adrenoceptor subtypes, see, Martin C. Michel, et al., Naunyn-
Schmiedeberg's Arch. Phacrmacol. (1995) 352:1-10.
The differences in the alpha adrenergic receptor subtypes
have relevance in pathophysiologic conditions. Benign prostatic
hyperplasia, also known as benign prostatic hypertrophy or BPH, is an
illness typically affecting men over fifty years of age, increasing in
severity with increasing age. The symptoms of the condition include,
but are not limited to, increased difficulty in urination and sexual
dysfunction. These symptoms are induced by enlargement, or
hyperplasia, of the prostate gland. As the prostate increases in size, it
impinges on free-flow of fluids through the male urethra.
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Concommitantly, the increased noradrenergic innervation of the
enlarged prostate leads to an increased adrenergic tone of the bladder
neck and urethra, further restricting the flow of urine through the
urethra.
In benign prostatic hyperplasia, the male hormone 5alpha-
dihydrotestosterone has been identified as the principal culprit. The
continual production of 5a-dihydrotestosterone by the male testes
induces incremental growth of the prostate gland throughout the life of
the male. Beyond the age of about fifty years, in many men, this
enlarged gland begins to obstruct the urethra with the pathologic
symptoms noted above.
The elucidation of the mechanism summarized above has
resulted in the recent development of effective agents to control, and in
many cases reverse, the pernicious advance of BPH. In the forefront of
these agents is Merck & Co., Inc's product PROSCAR~ (finasteride).
The effect of this compound is to inhibit the enzyme testosterone 5-a
reductase, which converts testosterone into 5a-dihydrotesterone,
resulting in a reduced rate of prostatic enlargement, and often reduction
in prostatic mass.
The development of such agents as PROSCAR~ bodes well
for the long-term control of BPH. However, as may be appreciated from
the lengthy development of the syndrome, its reversal also is not
immediate. In the interim, those males suffering with BPH continue to
suffer, and may in fact lose hope that the agents are working sufl'lciently
rapidly.
In response to this problem, one solution is to identify
pharmaceutically active compounds which complement slower-acting
therapeutics by providing acute relief. Agents which induce relaxation
of the lower urinary tract tissue, by binding to alpha 1 adrenergic
receptors, thus reducing the increased adrenergic tone due to the
disease, would be good candidates for this activity. Thus, one such agent
is alfuzosin, which is reported in EP 0 204597 to induce urination in
cases of prostatic hyperplasia. Likewise, in WO 92/0073, the selective
ability of the R(+) enantiomer of terazosin to bind to adrenergic receptors
of the alphal subtype was reported. In addition, in WO 92/161213,
combinations of 5a-reductase inhibitory compounds and alphal-
adrenergic receptor blockers (terazosin, doxazosin, prazosin, bunazosin,
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indoramin, alfuzosin) were disclosed. However, no information as to the
alpha 1d, alpha 1b, or alpha la subtype specificity of these compounds
was provided as this data and its relevancy to the t>"eatment of BPH was
not known. Current therapy for BPH uses existing non-selective alpha 1
antagonists such as prazosin (Minipress, Pfizer), Terazosin {Hytrin,
Abbott) or doxazosin mesylate (Cardura, Pfizer). These non-selective
antagonists suffer from side effects related to antagonism of the alpha ld
and alpha lb receptors in the peripheral vasculature, e.g., hypotension
and syncope.
The recent cloning of the human alpha la adrenergic
receptor (ATCC CRL 11140) and the use of a screening assay utilizing
the cloned human alpha la receptor enables identification of compounds
which specifically interact with the human alpha la adrenergic
receptor. [PCT International Application Publication Nos. W094/08040,
published 14 April 1994 and W094/10989, published 26 May 1994] As
disclosed in the instant patent disclosure, a cloned human alpha 1a
adrenergic receptor and a method for identifying compounds which bind
the human alpha la receptor has now made possible the identification of
selective human alpha 1a adrenergic receptor antagonists useful for
treating BPH. The instant patent disclosure discloses novel compounds
which selectively bind to the human alpha la receptor. These
compounds are further tested for binding to other human alpha 1
receptor subtypes, as well as counterscreened against other types of
receptors (e.g., alpha 2), thus defining the specificity of the compounds of
the present invention for the human alpha 1a adrenergic receptor.
It is an object of the present invention to identify compounds
which bind to the alpha la adrenergic receptor. It is a further object of
the invention to identify compounds which act as antagonists of the
alpha la adrenergic receptor. It is another object of the invention to
identify alpha la adrenergic receptor antagonist compounds which are
useful agents for treating BPH in animals, preferably mammals,
especially humans. Still another object of the invention is to identify -- -
alpha 1a adrenergic receptor antagonists which are useful for relaxing
lower urinary tract tissue in animals, preferably mammals, especially
humans.
It has now been found that the compounds of the present
invention are alpha la adrenergic receptor antagonists. Thus, the
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compounds of the present invention are useful for treating BPH in
mammals. Additionally, it has been found that the alpha 1a adrenergic
receptor antagonists of the present invention are also useful for relaxing
lower urinary tract tissue in mammals.
SUMMARY OF THE INVENTION
The present invention provides compounds for the
treatment of urinary obstruction caused by benign prostatic hyperplasia
(BPH). The compounds antagonize the human alpha la adrenergic
receptor at nanomolar and subnanomolar concentrations while
exhibiting at least ten fold lower afI~lnity for the alpha ld and alpha lb
human adrenergic receptors and many other G-protein coupled
receptors. This invention has the advantage over non-selective alpha 1
adrenoceptor antagonists of reduced side effects related to peripheral
adrenergic blockade. Such side effects include hypotension, syncope,
lethargy, etc. The compounds of the present invention have the
structure:
R24
M R~s R~s
/ m R
n E ) R2s
R l N C Q
I
R1 J ~ ~ Rs q
~G RI~R~s p
wherein (a is selected from
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\ ~X)s ~~~X)s '~~x)s
~ O vl ~
O v
v Ra ~ Ra
N R4 ~ N~ N Rs N~ N S
R ~ 1o R ~
R7 ~ ~ s O O R O~ R9
O H R , R1o
~x)s
//)
R R2~13 O \
iN 1 1 ~WX)s ~N NR$
p a , R~ ~ ~
R5 N O
H '
~x)s ~x)s
\ \
p \ ~ R11R12
N~ N W ~ N7 N I
R ~ R
O H 1~ R 11 , O N ~\
R H O
13
R R14 R13 14 p ~X)s
~~Y RsN R -N
O O ,
Z
Ra , 0 i _N or 0 S
O
E, G, L and M are each independently selected from hydrogen, C1-g
alkyl, C3-g cycloalkyl, (CH2)0-4086, (CH2)0-4N(Rl9)2, (CH2)0-4CN,
(CH2)p-4CF3, (CH2)0-4C02R19, (CH2)p_4CON(R19)2, (CH2)p-4S02RS or
(CH2)p_4S02N(Rl9)2;
J is selected from hydrogen, C1-g alkyl, C3-g cycloalkyl, (CH2)1-4086,
(CH2)1-4N(R19)2, (CH2)1-4CN, (CH2)p_4CF3~ (CH2)0-~C02R19,
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(CH2)0_4CON(Rl9)2~ (CH2)0-4S02R6, or (CH2)0-4S02N(R19)2~
R1 is selected from unsubstituted, mono- or poly-substituted phenyl
wherein the substituents on the phenyl are independently selected from
halogen, CF3, cyano, nitro, OR6, N(R19)2, NR19COR20,
NR19CON(R20)2~ NR19Sp2R6~ NR19S02N(R20)2~ (CH2)0-4C02R19,
(CH2)p_4CON(Rl9)2~ (CH2)0-4S02N(R19)2~ (CH2)0-4S02R6 or
C1_4 alkyl; or unsubstituted, mono- or poly-substituted pyridyl, pyridyl N-
oxide (NCO) pyrazinyl, thienyl, thiazolyl, furanyl, quinazolinyl or
naphthyl wherein the substituents on the pyridyl, pyrazinyl, thienyl,
thiazolyl, furanyl, quinazolinyl, or naphthyl are independently selected
from CF3, cyano, nitro, N(Rl9)2, (CH2)0-4C02R19~ (CH2)0-4CON(R19)2,
(CH2)0-4S02N(Rl9)2, (CH2)0-4S02R6~ phenyl, OR6, halogen, C1_4 alkyl
or C3_g cycloalkyl;
R is selected from hydrogen, cyano, OR6, C02R19, CON(R19)2, tetrazole,
isooxadiazole, unsubstituted, mono- or poly-substituted phenyl wherein
the substituents on the phenyl are independently selected from halogen,
cyano, nitro, OR6, (CH2)0-4C02R19~ (CH2)0-4CON(R19)2, N(R19)2,
NR19COR6, NR19CON(R20)2, NR.19S02R6, NR,19S02N(R20)2,
(CH2)0-4S~2N(R19)2, (CH2)0-4S02R6 or C1-4 alkyl; or unsubstituted,
mono- or poly-substituted pyridyl, thienyl, furanyl or naphthyl wherein
the substituents on the pyridyl, thienyl, furanyl or naphthyl are
independently selected from CF3, (CH2)0-4C02R19, (CH2)0_4CON(R19)2,
(CH2)0_4SO2N(Rl9)2, (CH2)0-4S~2R6~ Phenyl, OR6, halogen, C1_4 alkyl
or C3_g cycloalkyl;
R2~ R3 and R7 are each independently selected from hydrogen, C1-g
alkyl, C4-g cycloaikyl, (CH2)0-4C02R19~ (CH2)0-4CON(R19)2, (CH2)0-
4COR6, (CH2)2-4086, (CH2)1-4CF3> (CH2)0-4S02R6, (CH2)0-4S02N(R19)2
or (CH2)1-4CN;
R4 is selected from hydrogen, (CH2)0-4COR6, (CH2)0-4CN, (CH2)0-4CF3~
(CH2)0-4C02R19~ (CH2)0_4CON(R19)2~ (CH2)0-4S02R6 or
(CH2)0-4S02N(Rl9)2;
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R5, Rg, R10, R15~ R16~ R17 and R18 are each independently selected from
hydrogen, C1-g alkyl, C3-g cycloalkyl, (CH2)2-4086 or
(CH2)0-4CF3~
R6 is selected from hydrogen, C 1-g alkyl, Cg-g cycloalkyl or
(CH2)0-4CF3~
R9 is selected from hydrogen, C1-g alkyl, Cg-g cycloalkyl, C02R6,
CON(R6)2, (CH2)1-408,6 or (CH2)0_4CF3~
R11 and R12 are each independently selected from hydrogen,
C1-g alkyl or C3-g cycloalkyl;
R13 and R14 are each independently selected from hydrogen, C1-g alkyl,
C3-g cycloalkyl, (CH2)1-4086, (CH2)0-4CF3~ unsubstituted, mono- or
poly-substituted phenyl wherein the substituents on the phenyl are
independently selected from halogen, CF3, cyano, vitro, OR6,
(CH2)0-4CON(Rl9)2, (CH2)0-4C02R19 or C1-4 alkyl; or unsubstituted,
mono- or poly-substituted: pyridyl, thienyl, furanyl or naphthyl wherein
the substituents on the pyridyl, thienyl, furanyl or naphthyl are
independently selected from CFg, phenyl, OR6, halogen, C1-4 alkyl or
C3-g cycloalkyl;
R19 and R20 are each independently selected from hydrogen, C1-g alkyl,
Cg-g cycloalkyl or (CH2)1-4CF3;
R22 is selected from hydrogen, C1-g alkyl, Cg-g cycloalkyl, (CH2)0-4086
or (CH2)0-4CF3~
R24 and R26 are each independently selected from hydrogen or OR2g;
R28 is selected from hydrogen, C1_g alkyl, C3-g cycloalkyl or ~ -
(CH2)0-4CF3'>
W is O or NR11;
_g_
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each X is independently selected from halogen, cyano, vitro, C 1-g alkyl,
C3-g cycloalkyl, (CH2)0-4086 or (CH2)0-4CF3~
Y is C-R6 or N;
Z is hydrogen, oxygen or sulphur;
m, p and q are each independently an integer of from zero to two
provided that when q is zero, R2s is hydrogen;
n, o, s and t are each independently an integer of from zero to.four;
v is an integer from zero to one;
and the pharmaceutically acceptable salts thereof.
A first embodiment of the invention is a compound having
the structure
M R~5 R~s
1m 2
R l N C Q
1
R ~ J ~ ~R3 q
G RI~R~s P
wherein R1 is selected from unsubstituted, mono- or poly-substituted
phenyl wherein the substituents on the phenyl are independently
selected from halogen, CFg, cyano, vitro, OR6, N(Rlg)2, NR19COR20,
NRIgCON(R2~)2, NR1gS02R6, NR19S02N(R,20)2~ (CH2)0-4C02R1g,
(CH2)p_4CON(Rl9)2, (CH2)0_4S02N(Rl9)2,
(CH2)0-4S02R6 or C1_4 alkyl; or unsubstituted, mono- or poly-substituted
pyridyl, pyrazinyl, thienyl, thiazolyl, furanyl, quinazolinyl or naphthyl
wherein the substituents on the pyridyl, pyrazinyl, thienyl, thiazolyl,
furanyl, quinazolinyl, or naphthyl are independently selected from CF3,
cyano, vitro, N(Rlg)2, (CH2)0-4C02R1g, (CH2)0-4CON(Rl9)2,
(CH2)0-4S02N(Rlg)2, (CH2)0-4S02R6, Phenyl, OR6, halogen, C1-4 alkyl
or C3_g cycloalkyl;
R4 is selected from (CH2)0-4COR6, (CH2)0-4CN, (CH2)0-4CF3,
_g_
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(CH2)p_4C02R19, (CH2)0_4CON(Rl9)2, (CH2)0-4S02R6 or
(CH2)p_4SO2N(Rl9)2> ~d
R9 is selected from hydrogen, C1-g alkyl, C3-g cycloalkyl, (CH2)2-4086 or
(CH2)0-4CF3~
all other variables are as originally defined above; and the
pharmaceutically acceptable salts thereof.
In a second embodiment of the invention is the compound of
the formula
R24
Ris R1
m R
n E ~ R2 I
R l N C Q
R1 ~ 1~ ~s~p \R3 q
/o ~ ~ G R R
wherein
E, G, L, M and J are each independently selected from hydrogen,
C1-g alkyl, Cg-g cycloalkyl, or (CH2)0-4CF3~
R1 is selected from unsubstituted, mono-, di- or tri-substituted phenyl
wherein the substituents on the phenyl are independently selected from
halogen, CFg, cyano, nitro, OR6, N(R19)2, NR19COR20,
NR19CON(R20)2, NR19S02R6, NR,19S02N(R.20)2~ (CH2)0-4C02R19,
(CH2)0-4CON(R19)2, (CH2)0-4S02N(Rl9)2, (CH2)0-4S02R6 or
C1-4 alkyl; or unsubstituted, mono-, di- or tri-substituted pyridyl, pyridyl
N-oxide (N-~0), pyrazinyl, thienyl, thiazolyl, furanyl, quinazolinyl or
naphthyl wherein the substituents on the pyridyl, pyrazinyl, thienyl,
thiazolyl, furanyl, quinazolinyl or naphthyl are independently selected
from CF3, cyano, (CH2)0-4C02R19, (CH2)0-4CON(R19)2, (CH2)0-
4S02N(Rl9)2,
(CH2)0-4S02R6, phenyl, OR6, halogen, C1-4 alkyl or C3_g cycloalkyl;
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R is selected from hydrogen, cyano, OR6, C02R19, CON(R19)2, tetrazole,
isooxadiazole, unsubstituted, mono-, di- or tri-substituted phenyl
wherein the substituents on the phenyl are independently selected from
halogen, cyano, vitro, OR6, (CH2)0-4C02R19~ (CH2)0-4CON(R19)2,
N{Rl9)2~ NR19COR6, NR19CON(R20)2~ NR19s02R6~ NR19S02N{R20)2>
(CH2)0-4S02N(Rl9)2, (CH2)0_gS02R6 or C1-4 alkyl; or unsubstituted,
mono-, di- or tri-substituted pyridyl, thienyl, furanyl or naphthyl
wherein the substituents on the pyridyl, thienyl, furanyl or naphthyl are
independently selected from CF3, (CH2)0-4C02R19~ (CH2)0-4CON{R19)2,
(CH2)0-4S02N(Rl9)2, (CH2)0-4S02R6~ phenyl, ORS, halogen, C1-4 alkyl
or C3-g cycloalkyl;
R2, R3 and R7 are each independently selected from hydrogen, C 1-g
alkyl, C4-g cycloalkyl or (CH2)1-4CF3;
R13 and R14 are each independently selected from hydrogen, C1-g alkyl,
C3-g cycloalkyl, (CH2)1-4086, (CH2)0-4CF3~ substituted, mono-, di- or
tri-substituted phenyl wherein the substituents on the phenyl are
independently selected from halogen, CF3, cyano, vitro, OR6,
(CH2)0-4CON(R19)2, (CH2)0-4C02R19 or C1-4 alkyl; or unsubstituted,
mono-, di- or tri-substituted: pyridyl, thienyl, furanyl or naphthyl
wherein the substituents on the pyridyl, thienyl, furanyl or naphthyl are
independently selected from CF3, phenyl, OR6, halogen, C1-4 alkyl or
C3-g cycloalkyl; and
n and t are each independently an integer from zero to two;
and all other variables are as originally defined above [previously];
and the pharmaceutically acceptable salts thereof.
In a third embodiment of the invention is the compound of
formula
R1s Rts m 2 _
I
R N ~ C Q
R1 J )
q
G R~~R~B
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wherein R1 is selected from unsubstituted, mono-, di- or tri-substituted
phenyl wherein the substituents on the phenyl are independently
selected from halogen, CF3, cyano, nitro, OR6, N(R19)2, NR19COR20,
NR19CON(R20)2~ NR19S02R6~ NR19S02N(R20)2~ (CH2)0-4C02R19,
(CH2)0-4CON(Rl9)2~ (CH2)p_4S02N(Rl9)2~ (CH2)0-4SO2R6 or
C1-4 alkyl; or unsubstituted, mono-, di- or tri-substituted pyridyl,
pyrazinyl, thienyl, thiazolyl, furanyl, quinazolinyl or naphthyl wherein
the substituents on the pyridyl, pyrazinyl, thienyl, thiazolyl, furanyl,
quinazolinyl or naphthyl are independently selected from CFg, cyano,
(CH2)0_4C02R19, (CH2)0_4CON(Rl9)2~ (CH2)0_4S02N(R19)2~
(CH2)0-4S02R6, phenyl, OR6, halogen, C1-4 alkyl or C3-g cycloalkyl; and
all other variables are as defined in the first embodiment; and the
pharmaceutically acceptable salts thereof.
In a first class of the invention is the compound selected
from
R 24 R24
R1 N R2s R R2s
R 1 N~ (CH~q Q
(CH2)q Q , ,
R R24
R~ N ~ R2s R R2s
H2)-Q or R1 N~~(~H2~Q .
wherein C~,1 is selected from
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'~ X)s ~~ ~X)s \~ ~X)s
O ~ ~ O ~ O
R8 Ra
~~N~N R4 ~\N~ N R9 ~~N~N
R~ ~ ~ R ~ Rio R~ S
O N R 5 O O , O!~ 9 o r
H , R1o R
7 R2~13
R
iN ~ ~i~ (X)s
~t
O
R1 is selected from unsubstituted, mono-, di- or tri-substituted phenyl
wherein the substituents on the phenyl are independently selected from
halogen, CF3, cyano, nitro, OR6, (CH2)0-2C02R19, (CH2)0-2CON(R~9)2,
(CH2)0-2S02N(R19)2, (CH2)0-2S02R6 or C1-4 alkyl; or unsubstituted,
mono-, or di-substituted pyridyl or pyridyl N-oxide, wherein the
substituents on the pyridyl or pyridyl N-oxide are independently selected
from halogen, CF3, cyano, OR6,
(CH2)0_2C02R19, (CH2)0_2CON(Rl9)2~ (CH2)0_2S02N(Rl9)2~
(CH2)0-2S02R6 or C1-4 alkyl;
R is selected from hydrogen, cyano, ORS, C02R19, CON(R19)2, or
unsubstituted, mono- or di-substituted phenyl wherein the substituents
on the phenyl are independently selected from halogen, cyano, nitro,
OR6, (CH2)0-2C~2R.19~ (CH2)0-2CON(Rl9)2 or C1_4 alkyl;
R4 is selected from hydrogen, CORE, C02R19, S02R6 or CON(R19)2;
R~ is selected from hydrogen, C1-g alkyl, C3-g cycloalkyl, (CH2)0-3086 or
(CH2)0-3CF3~
R6 is selected from hydrogen, C1-g alkyl, C3-g cycloalkyl or
(CH2)0-3CF3~
Rg and R10 are each independently selected from hydrogen,
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C1-s alkyl, C3-g cycloalkyl, (CH2)2-4086 or (CH2)0-3CF3;
R9 is selected from hydrogen, C1-6 alkyl, C3-g cycloalkyl,
C02R6, CON{R6)2, (CH2)1-4086 or (CH2)0-3CF3~
R13 is selected from hydrogen, C1-6 alkyl, Cg-g cycloalkyl,
(CH2)2-4086, (CH2)0-2CF3 or unsubstituted, mono- or di-substituted
phenyl wherein the substituents on the phenyl are independently
selected from halogen, CF3, cyano, nitro, amino, OR6, C02R19 or C1-4
alkyl;
R19 is selected from hydrogen, C1-6 alkyl, C3-g cycloalkyl or
(CH2)1-3CF3~ ~d
R22 is selected from hydrogen, C1-g alkyl, C3-6 cycloalkyl, (CH2)0-4086
or {CH2)0-3CF3~
R24 and R26 are each independently selected from hydrogen or OR28,
wherein R2g is selected from hydrogen, C 1-6 alkyl, C3-g cycloalkyl or
(CH2)1-3CF3~
s is an integer from zero to three;
and all other variables are as defined above in the second embodiment;
and the pharmaceutically acceptable salts thereof.
In a second class of the invention is the compound selected
from
R (CH2rQ R
R~ N~ R~ N~(CH2}q Q
R (CH2rQ R
R NJ or R1 N~~(CH2~-Q .
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wherein Rl is selected from unsubstituted, mono-, di- or tri-substituted
phenyl wherein the substituents on the phenyl are independently
selected from halogen, CFg, cyano, nitro, OR6, (CH2)0-2C02R19, (CH2)0-
2CON(Rl9)2,
(CH2)0-2S02N(R19)2, (CH2)0-2S02R6 or C1_4 alkyl; or unsubstituted,
mono-, or di-substituted pyridyl wherein the substituents on the pyridyl
are independently selected from halogen, CF3, cyano, OR6,
(CH2)0-2C02R19~ (CH2)0-2CON(R19)2~ (CH2)0-2S02N(R,19)2~
(CH2)0-2S02R6 or C1-4 alkyl;
R4 is selected from CORE, C02R19, S02R6, or CON(R19)2; and
all other variables are as defined above in the third embodiment; and the
pharmaceutically acceptable salts thereof.
In a first subclass of the invention is the compound selected
from
R R24
\ N
R2s
(R2y~~~A (CH2~C,~ ,
R 24
R R2s
\ N (CH2~'Q
(R21 ~~~~'-A
R24
R
\ N~ R2s
(R2~~~~~A ~(CH2~q a _ _
or
R2s
R
\ N~~~(CH2}~-Q
U
(R2~~~~~A
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wherein 14,1 is selected from
\~X)s ~~~X)s .
I / O I ~ ~X)s
R4 ~~ N~ s
N N N R ~~ N \
H
H O~O or H ~ ~ ,
O N R R
H
R is selected from hydrogen, OR6 or cyano;
A is selected from C-R21 or N or N-~O;
R13 is selected from hydrogen, C1_4 alkyl or unsubstituted, mono- or di-
substituted phenyl wherein the substituents on the phenyl are
independently selected from halogen, CFg, cyano, nitro, amino, OR6,
C02R19 or C1_4 alkyl;
each X is a halogen;
each R21 is independently selected from hydrogen, halogen, hydroxy,
cyano, OC1_4 alkyl, OCF3, OCH2CF3, C02-C1_4 alkyl, CONH2, S02NH2
or S02C1_4 alkyl; and
R24 and R26 are each independently selected from hydrogen or OR28,
wherein R28 is selected from hydrogen, C1_4 alkyl, or (CH2)0_2CF3;
r is an integer from zero to two;
and all other variables are as defined in the first class above;
and the pharmaceutically acceptable salts thereof.
In a second subclass of the invention is the compound
selected from - -
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R
N
(R21~~~~A (CH2~-Q
R
N (CH2~-(~
/., ,
(R21~ ~'~A
R ,,,,(CH2rq--Q
NJ
U or
(R21~ ~'~A
R
N~~(CH2}~-Q
(R2~ ~ ~'-'A
wherein A is selected from C-R21 or N;
R13 is selected from hydrogen, C1_4 alkyl or unsubstituted, mono- or di-
substituted phenyl wherein the substituents on the phenyl are
independently selected from halogen, CF3, cyano, nitro, amino, OR6,
C02R19 or
C 1_4 alkyl;
each X is a halogen;
each R21 is independently selected from hydrogen, halogen, hydroxy,
cyano, OC1_4 alkyl, OCFg, OCH2CF3, C02-C1_4 alkyl, CONH2, S02NH2
or S02C1_4 alkyl;
r is an integer from zero to two;
17-
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all other variables are as defined in the second class above; and the
pharmaceutically acceptable salts thereof.
In a first illustration of the invention is the compound
wherein Q is selected from
F F
F \ F
I\ I, F
O ~ F
~~ N~N R4 ~~ N N ~ \
H ~ I H ~ ~ ~ N
O~N R5 O O or H R13
H
R is selected from hydrogen, hydroxy or cyano;
R4 is C02R19;
R5 is (CH2)0-3086;
q is an integer from zero to one;
and all other variables are as defined in the first subclass above;
and the pharmaceutically acceptable salts thereof; provided that the
compound is not (4S)-trc~ns-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-
carboxylic acid-[1-(4-hydroxy-4-pyridin-2-yl-cyclohexyl)-(3R)-pyrrolidin-3-
yl]amide.
In a second illustration of the invention is the compound
wherein Q is selected from
F F
F \ F
I \ ~ / F
O ~ F
~\ N " N R4 ~\ N N
H ~~ ~
H O~ N R5 O p or H R~3 ,
H
R is selected from hydrogen, hydroxy or cyano;
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R4 is C02R19;
R5 is (CH2)0-3086;
q is an integer from zero to one;
and all other variables are as defined in the second subclass above;
and the pharmaceutically acceptable salts thereof; provided that the
compound is not (4S)-tracns-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-
carboxylic acid-[1-(4-hydroxy-4-pyridin-2-yl-cyclohexyl)-(3R)-pyrrolidin-3-
yl]amide.
In a third illustration of the invention is the compound
wherein
R is H or OH;
Q is selected from
F F
\ F I \ F
O
R4 Of ~~ 9
~~ N-C~N -C. R
H ~ I s H N O
O N R O
H
R4 is H or C02CH3;
R5 is H, CH3, or CH20CH3;
R9 is H, CHg, cyclopropyl, CONH2, CH20H, or COOCH3;
q is an integer from zero to one; and all other variables are as defined in
the first subclass above; and pharmaceutically acceptable salts thereof;
provided that the compound is not (4S)-trans-4-(3,4-difluorophenyl)-2-
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oxooxazolidine-3-carboxylic acid-[1-(4-hydroxy-4-pyridin-2-yl-cyclohexyl)-
(3R)-pyrrolidin-3-yl]amide hydrochloride.
One aspect of the invention is the compound having the
formula:
R
R ~ N~
wherein
R is H or OH;
R1 is suitably unsubstituted pyridyl, unsubstituted pyridyl N-oxide,
unsubstituted phenyl, or mono- or poly-substituted phenyl; typically
unsubstituted pyridyl, unsubstituted pyridyl N-oxide, unsubstituted
phenyl, or mono- or di-substituted phenyl; more typically 2-pyridyl, 2-
pyridyl N-oxide, 4-substituted phenyl, 2-substituted phenyl, or 2,4-
substituted phenyl; wherein the phenyl substituents are independently
selected from fluorine, cyano, OH, OCHg, C02CH2CH3, and CF3; and
Q is
F F
\ F I \ F
O
R4 °r n
~ N~ ~N ~ ,C,N Rs
H ,
O I s H ~O
R O
or more preferably is
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F F
\ F I \ F
O
w .C. R4 or
N N ~~N.C~N
H
O I Rs H ~-O
N O
H
wherein
R4 is H or C02CH3;
R5 is H, CH3, or CH20CH3; and
R9 is H, CH3, cyclopropyl, CONH2, CH20H, or COOCH3; provided that
the compound is not (4S)-trams-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-
carboxylic acid-[1-(4-hydroxy-4-pyridin-2-yl-cyclohexyl)-(3R)-pyrrolidin-3-
yl]amide hydrochloride.
Exemplifying the invention is the compound selected from
(4S)-3-{1-[4-(2-cyano-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl)-4-
(3,4-difluoro-phenyl)-6-methyoxymethyl-2-oxo-1,2,3,4-tetra
hydropyrimidine-5-carboxylic acid methyl ester;
(4S)-cis-3-{1-[4-(2-cyano-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl}-4-(3,4-difluoro-phenyl)-6-methyoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester;
trams-4S-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4
(2-cyano-phenyl)-cyclohexyl]-3R-pyrrolidin-3-yl}amide;
(4S)-cis-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4-(2
cyano-phenyl)-cyclohexyl]-(3R)--pyrrolidin-3-yl}amide;
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(4S)-trans-4-(3,4-difluoro-phenyl)-3-{1-[4-(2-ethoxycarbonyl-phenyl)-
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo-
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester;
(4S)-cis-4-(3,4-difluoro-phenyl)-3-{1-[4-(2-ethoxycarbonyl-phenyl)-
cyclohexyl]-(3R)--pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester;
(4S)-tr~ns-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid-{1-[4-
(2-ethoxycarbonylphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide;
(4S)-cis-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4-(2
ethoxycarbonylphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide;
(4S)-trans-4-(3,4-difluorophenyl)-3-[1-(4-pyridin-2-yl-cyclohexyl)-(3R)-
pyrrolidin-3-ylcarbamoyl]-S-methoxymethyl-2-oxo-1,2,3,4-tetrahydro
pyrimidine-5-carboxylic acid methyl ester;
(4S)-cis-4-(3,4-difluorophenyl)-3-[1-(4-pyridin-2-yl-cyclohexyl)-(3R)-
pyrrolidin-3-ylcarbamoyl)-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester;
(4S)-trans-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid-[1-(4
pyridin-2-yl-cyclohexyl)-(3R)-pyrrolidin-3-yl]amide;
(4S)-cis-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-[1-(4
pyridin-2-yl-cyclohexyl)-(3R)-pyrrolidin-3-yl]amide;
( 4S )-trot ns-4-(3, 4-difluorophenyl )-6-methoxymethyl-3- { 1-[4-( 2-methoxyl-
phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-2-oxo-1,2,3,4-
tetrahydro-pyrimidine-5-carboxylic acid methyl ester;
(4S)-cis-4-(3,4-difluoro-phenyl)-6-methoxymethyl-3-{1-[4-(2-methoxyl
phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-2-oxo-1,2,3,4
tetrahydro-pyrimidine-5-carboxylic acid methyl ester;
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(4S)-trczns-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4
(2-methoxyphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide;
(4S)-cis-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4-(2-
methoxyphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide;
(4S)-cis-4-(3,4-difluoro-phenyl) -3-[1-(4-hydroxy-4-pyridin-2-yl-cyclohexyl)
(3R)-pyrrolidin-3-ylcarbamoyl]-6-methoxymethyl -2-oxo-1,2,3,4
tetrahydro-pyrimidine-5-carboxylic acid methyl ester;
(4S)-traps-4-(3,4-difluoro-phenyl) -3-[1-(4-hydroxy-4-pyridin-2-yl
cyclohexyl)-(3R)--pyrrolidin-3-ylcarbamoyl]-6-methoxymethyl -2-oxo
1,2,3,4-tetrahydro-pyrimidine-5-carboxylic acid methyl ester;
(4S)-cis-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-[1-(4-
hydroxy-4-pyridin-2-yl-cyclohexyl)-(3R)--pyrrolidin-3-yl]amide;
(4S)-4-(3,4-difluoro-phenyl)-3-{ 1-[4-(2-hydroxy-phenyl)-cyclohexyl]-(3R)-
pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester;
( 4S )-cis-4-( 3,4-difluoro-phenyl )-6-methoxymethyl-3- { 1-[4-( 2-hydro xy-
phenyl)-cyclohexyl]-(3R)--pyrrolidin-3-ylcarbamoyl}-2-oxo-1,2,3,4-
tetrahydro-pyrimidine-5-carboxylic acid methyl ester;
(4S)-traps-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4
(2-hydroxyphenyl)-cyclohexyl]-(3R)--pyrrolidin-3-yl}amide;
(4S)-cis-4-(3,4-difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4-(2-
hydroxyphenyl)-cyclohexyl]-(3R)--pyrrolidin-3-yl}amide; and
the pharmaceutically acceptable salts thereof. - -
Also exemplifying the invention is the compound selected
from
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(4S )-tra ns-4-( 3-,4-difluoro-phenyl)-3- ( 1-[4-(2-fluoro-phenyl )-
cyclohexyl]-
(3R)-pyrrolidin-3-ylcarbamoyl)-6-methoxymethyl-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester;
b (4S)-traps-4-(3,4-difluoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid -(1-
[4-(2-fluoro-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}-amide;
(4S)-traps-4-(3,4-difluoro-phenyl)-3-(1-[4-(4-fluoro-2-methoxy-phenyl)-
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbau~oyl)-6-methoxymethyl-2-oxo-
1,2,3,4-tetrahydro-pyrimidine-5-carboxylic acid methyl ester;
(4S)-traps-4-(3,4-difluoro-phenyl)-2-oxo-oxazolidine-3-carboxylic arid (1-
[4-(4-fluoro-2-methoxy-phenyl ~cyclohexyl)-(3R)-pyrrolidin-3-yl }-amide;
(4S)-4-(3,4-difluoro-phenyl)-3-(1-[4-(2-fluoro-phenyl)-4-hydroxy-
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-1,2,3,4-
tetrahydro-pyrimidine-5-carboxylic acid methyl ester;
(4S)-4-(3,4-difluoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid -(1-[4-(2-
fluoro-phenyl)-4-hydroxy-cyclohexyl)-(3R)-pyrrolidin-3-yll-amide;
(4S)-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid (1-[4-
hydroxy-4-( 2-trifluoromethylphenyl )-cycl ohexyl]-( 3R )-pyrrolidia-3-yl }
amide;
(4S)-4-(3,4-difluorophenyl)-3-~ 1-[4-hydroxy-4-(2-trifluoromethylphenyl)-
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyll-6-methoxymethyl-2-oxo-
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester;
SO (4S)-4-(3,4-difluorophenyI)-2-oxo-oxazotidine-3-carboxylic acid (1-[4-(2
trifluoromethylphenyl)-cycIohexyl]-(3R)-pyrrolidin-3-yl} amide;
(4S)-4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-3-(1-[4-(2- _ __
trifluoromethylphenyl)-cyclohexyl)-(3R)-pyrrolidin-3-ylcarbamoyl}-
1,2,3,4-tetrahydropyri-midine-5-carboxylic acid methyl ester;
(4S)~4-(3,4-difluorophenyl)-3-(1-[4-(4-fluorophenyl)-4-hydroxy-cyclohexyl]-
_2g-
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(3R)-pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester;
(4S)-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid{1-[4-(4-
fluorophenyl)-4-hydroxy-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide;
(4S)-4 -(3,4-difluorophenyl)-3-{1-[4-(4-fluorophenyl)-4-hydroxy-cyclohexyl]-
(3R)-pyrrolidin-3-ylcarbamoyl}-6-methyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester;
(4S)-4 -(3,4-difluorophenyl)-3-{1-[4-(4-fluorophenyl)-cyclohexyl]-(3R)-
pyrrolidin-3-ylcarbamoyl}-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-
carboxylic acid methyl ester;
(4S)-4 -(3,4-difluorophenyl)-3-{1-[4-(4-fluorophenyl)-cyclohexyl]-(3R)-
pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester;
(4S)-4 -(3,4-difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid{1-[4-(4-
fluorophenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide;
(4S)-4 -(3,4-difluorophenyl)-3-{1-[4-(4-fluorophenyl)-4-hydraxy-cyclohexyl]-
(3R)-pyrrolidin-3-ylcarbamoyl}-2-oxo-1,2,3,4-tetrahydropyrimidine-5-
carboxylic acid methyl ester;
(4S)-3-{1-[4-(4-cyanophenyl)-4-hydroxycyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl}-4 -(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester;
5-cyclopropyl-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid
{ 1-[4-(2-fluorophenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl-amide;
(4S, 5S)-5-cyclopropyl-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-
carboxylic acid{1-[4-(4-fluorophenyl)-4-hydroxy-cyclohexyl]-(3R)-
pyrrolidin-3-yl}amide;
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(4S, 5S)-5-cyclopropyl-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-
carboxylic acid(1-[4-(4-cyanophenyl)-cyclohexyl]-(3R)-pyrrolidin-3-
yl} amide;
(4S)-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid{1-[4-(4
cyanophenyl)-4-hydroxy-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide;
(4S)-3-{1-[4-(4-cyanophenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-4-
(3,4-difluorophenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-
carboxylic acid methyl ester;
(4S)-trans-4-(3,4-difluorophenyl)-3-[1-(4-pyridin-2-yl-cyclohexyl)-(3R)
pyrrolidin-3-ylcarbamoyl]-6-methyl-2-oxo-1,2,3,4-tetrahydro-pyrimidine
5-carboxylic acid methyl ester;
(4S)-3-{1 -[4-(2-cyano-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3- ylcarbamoyl}-
4-(3,4-difluoro-phenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydro-pyrimidine-5-
carboxylic acid methyl ester;
(4S)-3-{1 -[4-(2-cyano-4-fluoro-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl}-4-(3,4-difluoro-phenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydro-pyrimidine-5-carboxylic acid methyl ester;
(4S)-4-(3,4-difluoro-phenyl)-3-{1 -[4-(4-fluoro-2-hydroxy-phenyl)-
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo
1,2,3,4-tetrahydro-pyrimidine-5-carboxylic acid methyl ester;
(4S)-4-(3,4-difluoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid {1-[4 -(4-
fluoro-2-hydroxy-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3- yl}-amide;
(4S)-4-(3,4-difluoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid {1-[4 -(2-
cyano-4-fluoro-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}-amide; ~ --
(4S)-4-(3,4-difluoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid {1-[4 -(2-
cyano-4-fluoro-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}-amide;
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(4S,5S)-4-(3,4-difluoro-phenyl)-5-methyl-2-oxo-oxazolidine-3-carboxylic
acid {1-[4 -(2-cyano-4-fluoro-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}-
amide;
(4S,5S)-5-Cyclopropyl-4-(3,4-difluoro-phenyl)-2-oxo-oxazolidine-3-
carboxylic acid {1-[4 -(2-cyano-4-fluoro-phenyl)-cyclohexyl]-(3R)-
pyrrolidin-3y1}-amide;
(4S)-4-(3,4-difluoro-phenyl)-3-{1-(4-(2-fluoro-phenyl)-4-hydroxy-cyclohex-1-
yl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6-methyl-2-oxo-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester;
(4S)-trams-4-(3,4-difluoro-phenyl)-3-{ 1-(4-(2-fluoro-phenyl)-cyclohexyl]-
(3R)-pyrrolidin-3-ylcarbamoyl}-6-methyl-2-oxo-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester;
(4S)-cis-4-(3,4-difluoro-phenyl) -3-[1-(4-hydroxy-4-pyridin-2-yl-cyclohexyl)-
(3R)-pyrrolidin-3-ylcarbamoyl] -2-oxo-1,2,3,4-tetrahydro-pyrimidine-5-
carboxylic acid methyl ester;
(4S)-tracns-4-(3,4-difluorophenyl)-3-[1-(4-pyridin-2-yl-cyclohexyl)-(3R)-
pyrrolidin-3-ylcarbamoyl]-2-oxo-1,2,3,4-tetrahydro-pyrimidine-5-
carboxylic acid methyl ester;
(4S)-trc~ns-3{1-[4-(2-cyano-phenyl)-piperidin-1-yl]-(3R)-pyrrolidin-3-
ylcarbamoly}-4-(3,4-difluoro-phenyl)-2-oxo-1,2,3,4-tetrahydro-pyrimidine-
5-carboxylic acid methyl ester;
(4S,5S)-tracns-5-cyclopropyl-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-
carboxylic acid {1-[ 4-(4-fluorophenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl-
amide;
(4S)-4-(3,4-difluoro-phenyl) -3-[1-(4-hydroxy-4-pyridin-2-yl-cyclohexyl)
(3R)-pyrrolidin-3-ylcarbamoyl]-6-methyl -2-oxo-1,2,3,4-tetrahydro
pyrimidine-5-carboxylic acid methyl ester;
-2?-
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(4S)-traps-4-{3,4-difluorophenyl)-5-methyl-2-oxooxazolidine-3-carboxylic
acid-[1-(4-pyridin-2-yl-cyclohexyl)-{3R)-pyrrolidin-3-yl]amide;
traps-4S-(3,4-difluorophenyl)-3-[1-(4-oxopyridin-2-yl-cyclohexyl)-3R-
pyrrolidin-3-ylcarbamoyl)-6-methyl-2-oxo-1,2,3,4-tetrahydro-pyrimidine-
5-carboxylic acid methyl ester;
traps-2-(3,4-difluorophenyl)-1-[1-(4-pyridin-2-yl-cyclohexyl)-3R-
pyrrolidin-3-ylcarbamoyl]-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydro-
pyrimidine;
(4S,5R)-traps-4-(3,4-difluorophenyl-3-~1-[4-(4-fluoro-2-
methoxycarbonylphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-2-
oxo-oxazolidine-5-carboxylic acid methyl ester;
(4S,5R)-traps-4-(3,4-difluorophenyl)-3-( 1-{4-[4-fluoro-2-(2,2,2-
trifluoroethoxy)phenyl]cyclohexyl}-(3R)-pyrrolidin-3-ylcarbamoyl)-2-oxo-
oxazolidine-5-carboxylic acid methyl ester;
(4S,5R)-traps-4-(3,4-difluorophenyl)-2-oxo-3-[1-(4-pyridin-2-ylcyclohexyl)
(3R)-pyrrolidin-3-ylcarbamoyl]-oxazolidine-5-carboxylic acid methyl
ester;
(4S,5R)-traps-4-(3,4-difluorophenyl)-3-( 1-[4-(4-fluoro-2-
methoxyphenyl)cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl]-2-oxo-
oxazolidine-5-carboxylic acid methyl ester;
(4S,5R)-3-{1-[4-Cyano-4-(2-methoxyphenyl)cyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl}-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-5-carboxylic acid
methyl ester;
(4S,5R)-3-{1-[4-Cyano-4-(2-fluorophenyl)cyclohexyl]-(3R)-pyrrolidin-3-- -
ylcarbamoyl}-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-5-carboxylic acid
methyl ester;
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(4S,5R)-trans-3-{ 1-[4-( 2-cyanophenyl)cyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl}-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-5-carboxylic acid
methyl ester;
(4S,5R)-trans-4-(3,4-difluorophenyl)-5-hydroxymethyl-2-oxo-oxazolidine-
3-carboxylic acid {1-[4-(4-fluorophenyl)cyclohexyl]-(3R)-pyrrolidin-3-yl}
amide;
(4S,5R)-trans-4-(3,4-difluorophenyl)-5-hydroxymethyl-2-oxo-oxazolidine-
3-carboxylic acid {1-[4-(4-fluoro-2-methoxyphenyl)cyclohexyl]-(3R)-
pyrrolidin-3-yl} amide;
(4S,5R)-trans-4-(3,4-difluorophenyl)-3-( 1-[4-(4-fluoro-2-
methoxyphenyl)cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl]-2-oxo-
oxazolidine-5-carboxamide;
(4S,5R)-trans-4-(3,4-difluorophenyl)-3-{1-[4-(4-fluorophenyl)cyclohexyl]-
(3R)-pyrrolidin-3-ylcarbamoyl]-2-oxo-oxazolidine-5-carboxamide;
(4S,5R)-trans-4-(3,4-difluorophenyl)-3-{1-[4-(2-fluorophenyl)cyclohexyl]
(3R)-pyrrolidin-3-ylcarbamoyl]-2-oxo-oxazolidine-5-carboxamide;
(4S,5R)-trans-3-{1-[4-(2-cyano-4-fluorophenyl)cyclohexyl]-(3R)-pyrrolidin-
3-ylcarbamoyl}-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-5-carboxamide;
and
the pharmaceutically acceptable salts thereof.
In a preferred embodiment the compound is selected from
Compound A:
F
~ F __
F ~ ~ N O i
HO N ~ N "",
H
O~ O
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Compound B:
F
F
O
.,,N~. ~ co2nne
y'"'~ H ~. I
N O N
H
Compound C:
F
F
'"' t
N H O
~,,,~--l~N C OZMa
O
O~N Me
H
and the pharmaceutically acceptable salts thereof.
Another preferred embodiment is Compound A or a
pharmaceutically acceptable salt thereof. Another preferred
embodiment is Compound B or a pharmaceutically acceptable salt
thereof Still another preferred embodiment is Compound C or a
pharmaceutically acceptable salt thereof.
An illustration of the invention is a pharmaceutical
composition comprising a therapeutically e$'ective amount of any of the
compounds described above and a pharmaceutically acceptable carrier.
An example of the invention is a pharmaceutical composition made by
combining any of the compounds described above and a - -
pharmaceutically acceptable carrier. Another illustration of the
invention is a process for making a pharmaceutical composition
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comprising combining any of the compounds described above and a
pharmaceutically acceptable carrier.
Another example of the invention is the composition further
comprising a therapeutically effective amount of a testosterone 5-alpha
reductase inhibitor. Preferably, the testosterone 5-alpha reductase
inhibitor is a type 1, a type 2, both a type 1 and a type 2 (i.e., a three
component combination comprising any of the compounds described
above combined with both a type 1 testosterone 5-alpha reductase
inhibitor and a type 2 testosterone 5-alpha reductase inhibitor) or a dual
type 1 and type 2 testosterone 5-alpha reductase inhibitor. More
preferably, the testosterone 5-alpha reductase inhibitor is a type 2
testosterone 5-alpha reductase inhibitor. Most preferably, the
testosterone 5-alpha reductase inhibitor is finasteride.
More specifically illustrating the invention is a method of
treating benign prostatic hyperplasia in a subject in need thereof which
comprises administering to the subject a therapeutically effective
amount of any of the compounds (or any of the compositions) described
above.
Further exemplifying the invention is the method of
treating BPH wherein the compound (or composition) additionally does
not cause a fall in blood pressure at dosages effective to alleviate BPH.
Another illustration of the invention is the method of
treating benign prostatic hyperplasia wherein the compound is
administered in combination with a testosterone 5-alpha reductase
inhibitor. Preferably, the testosterone 5-alpha reductase inhibitor is
finasteride.
Further illustrating the invention is a method of inhibiting
contraction of prostate tissue or relaxing lower urinary tract tissue in a
subject in need thereof which comprises administering to the subject a
therapeutically effective amount of any of the compounds (or any of the
compositions) described above.
More specifically exemplifying the invention is the method
of inhibiting contraction of prostate tissue or relaxing lower urinary
tract tissue wherein the compound (or composition) additionally does not
cause a fall in blood pressures at dosages effective to inhibit contraction
of prostate tissue.
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More particularly illustrating the invention is the method of
inhibiting contraction of prostate tissue or relaxing lower urinary tract
tissue wherein the compound (or composition) is administered in
combination with a testosterone 5-alpha reductase inhibitor; preferably,
the testosterone 5-alpha reductase inhibitor is finasteride.
More particularly exemplifying the invention is a method of
treating a disease which is susceptible to treatment by antagonism of the
alpha 1a receptor which comprises administering to a subject in need
thereof an amount of any of the compounds described above effective to
treat the disease. Diseases which are susceptible to treatment by
antagonism of the alpha la receptor include, but are not limited to, BPH,
high intraocular pressure, high cholesterol, impotency, sympathetically
mediated pain, migraine (see, K.A. Vatz, Headache 1997:37: 107-108) and
cardiac arrhythmia.
An additional illustration of the invention is the use of any
of the compounds described above in the preparation of a medicament
for: a) the treatment of benign prostatic hyperplasia; b) relaxing lower
urinary tract tissue; or c) inhibiting contraction of prostate tissue; in a
subject in need thereof.
An additional example of the invention is the use of any of
the alpha la antagonist compounds described above and a 5-alpha
reductase inhibitor for the manufacture of a medicament for: a) treating
benign prostatic hyperplasia; b) relaxing lower urinary tract tissue; or c)
inhibiting contraction of prostate tissue which comprises an effective
amount of the alpha 1a antagonist compound and an effective amount of
5-alpha reductase inhibitor, together or separately.
DETAILED DESCRIPTION OF THE INVENTION
Representative compounds of the present invention exhibit
high selectivity for the human alpha la adrenergic receptor. One
implication of this selectivity is that these compounds display selectivity
for lowering intraurethral pressure without substantially affecting - -
diastolic blood pressure.
Representative compounds of this invention display
submicromolar af~lnity for the human alpha la adrenergic receptor
subtype while displaying at least ten-fold lower affinity for the human
alpha ld and alpha lb adrenergic receptor subtypes, and many other G-
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protein coupled human receptors. Particular representative compounds
of this invention exhibit nanomolar and subnanomolar affinity for the
human alpha la adrenergic receptor subtype while displaying at least 30
fold lower affinity for the human alpha ld and alpha lb adrenergic
receptor subtypes, and many other G-protein coupled human receptors
(e.g., serotonin, dopamine, alpha 2 adrenergic, beta adrenergic or
muscarinic receptors).
These compounds are administered in dosages effective to
antagonize the alpha la receptor where such treatment is needed, as in
BPH. For use in medicine, the salts of the compounds of this invention
refer to non-toxic "pharmaceutically acceptable salts." Other salts may;
however, be useful in the preparation of the compounds according to the
invention or of their pharmaceutically acceptable salts. Suitable
pharmaceutically acceptable salts of the compounds of this invention
include acid addition salts which may, for example, be formed by mixing
a solution of the compound according to the invention with a solution of a
pharmaceutically acceptable acid such as hydrochloric acid, sulphuric
acid, fumaric acid, malefic acid, succinic acid, acetic acid, benzoic acid,
citric acid, tartaric acid, carbonic acid or phosphoric acid.
Furthermore, where the compounds of the invention carry an acidic
moiety, suitable pharmaceutically acceptable salts thereof may include
alkali metal salts, e.g. sodium or potassium salts; alkaline earth metal
salts, e.g. calcium or magnesium salts; and salts formed with suitable
organic ligands, e.g. quaternary ammonium salts. Thus,
representative pharmaceutically acceptable salts include the following:
Acetate, Benzenesulfonate, Benzoate, Bicarbonate,
Bisulfate, Bitartrate, Borate, Bromide, Calcium, Camsylate, Carbonate,
Chloride, Clavulanate, Citrate, Dihydrochloride, Edetate, Edisylate,
Estolate, Esylate, Fumarate, Gluceptate, Gluconate, Glutamate,
Glycollylarsanilate, Hexylresorcinate, Hydrabamine, Hydrobromide,
Hydrochloride, Hydroxynaphthoate, Iodide, Isothionate, Lactate,
Lactobionate, Laurate, Malate, Maleate, Mandelate, Mesylate, - --
Methylbromide, Methylnitrate, Methylsulfate, Mucate, Napsylate,
Nitrate, N-methylglucamine ammonium salt, Oleate, Pamoate
(Embonate), Palmitate, Pantothenate, Phosphate/diphosphate,
Polygalacturonate, Salicylate, Stearate, Sulfate, Subacetate, Succinate,
Tannate, Tartrate, Teoclate, Tosylate, Triethiodide and Valerate.
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Compounds of this invention are used to reduce the acute
symptoms of BPH. Thus, compounds of this invention may be used
alone or in conjunction with a more long-term anti-BPH therapeutics,
such as testosterone 5-a reductase inhibitors, including PROSCAR~
(finasteride). Aside from their utility as anti-BPH agents, these
compounds may be used to induce highly tissue-specific, localized alpha
1a adrenergic receptor blockade whenever this is desired. Effects of this
blockade include reduction of intra-ocular pressure, control of cardiac
arrhythmias, and possibly a host of alpha la receptor mediated central
nervous system events.
The present invention includes within its scope prodrugs of
the compounds of this invention. In general, such prodrugs will be
functional derivatives of the compounds of this invention which are
readily convertible in vivo into the required compound. Thus, in the
methods of treatment of the present invention, the term "administering"
shall encompass the treatment of the various conditions described with
the compound specifically disclosed or with a compound which may not
be specifically disclosed, but which converts to the specified compound in
vivo after administration to the patient Conventional procedures for the
selection and preparation of suitable prodrug derivatives are described,
for example, in "Design of Prodrugs," ed. H. Bundgaard, Elsevier, 1985.
Metabolites of these compounds include active species produced upon
introduction of compounds of this invention into the biological milieu.
Where the compounds according to the invention have at
least one chiral center, they may accordingly exist as enantiomers.
Where the compounds according to the invention possess two or more
chiral centers, they may additionally exist as diastereoisomers. It is to
be understood that all such isomers and mixtures thereof are
encompassed within the scope of the present invention. Furthermore,
some of the crystalline forms for compounds of the present invention
may exist as polymorphs and as such are intended to be included in the
present invention. In addition, some of the compounds of the present -
invention may form solvates with water (i.e., hydrates) or common
organic solvents. Such solvates are also encompassed within the scope
of this invention.
The term "alkyl" shall mean straight or branched chain
alkanes of one to ten total carbon atoms, or any number within this
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range (i.e., methyl, ethyl, 1-propyl, 2-propyl, n-butyl, s-butyl, t-butyl,
etc.).
The term "alkenyl" shall mean straight or branched chain
alkenes of two to ten total carbon atoms, or any number within this
range.
The term "aryl" as used herein, except where otherwise
specifically defined, refers to unsubstituted, mono- or poly-substituted
aromatic groups such as phenyl or naphthyl.
The term "cycloalkyl" shall mean cyclic rings of alkanes of
three to eight total carbon atoms (i.e., cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl).
Whenever the term "alkyl" or "aryl" or either of their prefix
roots appear in a name of a substituent (e.g., aralkoxyaryloxy) it shall be
interpreted as including those limitations given above for "alkyl" and
"aryl." Designated numbers of carbon atoms (e.g., C1-10) shall refer
independently to the number of carbon atoms in an alkyl or cyclic alkyl
moiety or to the alkyl portion of a larger substituent in which alkyl
appears as its prefix root.
The term "halogen" shall include iodine, bromine, chlorine
and fluorine.
The term "substituted" shall be deemed to include multiple
degrees of substitution by a named substituent. The term "poly-
substituted" as used herein shall include di-, tri-, tetra- and penta-
substitution by a named substituent. Preferably, a poly-substituted
moiety is di-, tri- or tetra-substituted by the named substituents, most
preferably, di- or tri-substituted.
It is intended that the definition of any substituent or
variable (e.g., X, Rs, Rl9) at a particular location in a molecule be
independent of its definitions elsewhere in that molecule. Thus, -
N(R19)2 represents -NH2, -NHCH3, -NHC2H5, -N(CH3)C2H5, etc. and
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R2a
~_~~m represents for m=2
~(s~
OH OH OH OH
s~ , i .r~~ ~ .r'v , etc.
,
It is understood that substituents and substitution patterns on the
compounds of the instant invention can be selected by one of ordinary
skill in the art to provide compounds that are chemically stable and that
can be readily synthesized by techniques known in the art as well as
those methods set forth below. Where multiple substituent moieties are
disclosed or claimed, the substituted compound can be independently
substituted by one or more of the disclosed or claimed substituent
moieties, singly or plurally.
The term "Z is hydrogen," when refering to the "Q" group
R13
R1a
~Y
O
Z N
R8
refers to the moiety
R13
R1a
~Y
O
N
R8
The term heterocycle or heterocyclic ring, as used herein,
represents an unsubstituted or substituted stable 5- to 7-membered
monocyclic ring system which may be saturated or unsaturated, and
which consists of carbon atoms and from one to three heteroatoms
selected from N, O or S, and wherein the nitrogen and sulfur
heteroatoms may optionally be oxidized, and the nitrogen heteroatom
may optionally be quaternized. The heterocyclic ring may be attached at
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any heteroatom or carbon atom which results in the creation of a stable
structure. Examples of such heterocyclic groups include, but is not
limited to, piperidinyl, piperazinyl, oxopiperazinyl, oxopiperidinyl,
oxopyrrolidinyl, oxoazepinyl, azepinyl, pyrrolyl, pyrrolidinyl, furanyl,
thienyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl,
imidazolidinyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl,
oxazolidinyl, isooxazolyl, isoxazolidinyl, morpholinyl, thiazolyl,
thiazolidinyl, isothiazolyl, thiadiazolyl, tetrahydropyranyl,
thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone,
and oxadiazolyl. Morpholino is the same as morpholinyl.
The term "thienyl," as used herein, refers to the group
S
The terms "(+)-DHP" and "DHP" as used herein, refer to a
dihydropyrimidinone group of the formula
(X)q
O
~N R4
O' _N R5
H
for example:
F
F
O ~ O
~~N O _ _.
~ O
O' _ N
H
The term "activated (+)-DHP," as used herein, refers to a N-
3-(activated)carbamate of the desired dihydropyrimidinone where the
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activating group is, for example, a p-nitrophenyloxy group. A specific
example of an activated (+)-DHP is 4-(3,4-difluorophenyl)-5-
methoxycarbonyl-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydropyrimidine-
3-carboxylic acid (4-nitrophenyl ester), also referred to as the compound
15.
The term "(S)-oxa" as used herein, refers to an
oxazolidinone group of the formula
~x)q
/
R8 Rs
fit, N ~Rlo
-O
O
for example,
F
F
(
O
~t,~ N
--O
O
The term "activated (S)-oxa" as used herein, refers to an N-
(activated)carbamate of the desired oxazolidinone where the activating
group is, for example, a p-nitrophenyloxy group. A specific example of
an activated (S)-oxa group is 4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-
carboxylic acid 4-nitrophenyl ester (i.e., compound 16).
The term "lower urinary tract tissue," as used herein,
refers to and includes, but is not limited to, prostatic smooth muscle the
prostatic capsule, the urethra and the bladder neck.
The term "subject," as used herein refers to an animal,
preferably a mammal, most preferably a human, who has been the
object of treatment, observation or experiment.
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The term "therapeutically effective amount" as used
herein means that amount of active compound or pharmaceutical
agent that elicits the biological or medicinal response in a tissue,
system, animal or human that is being sought by a researcher,
veterinarian, medical doctor or other clinician, which includes
alleviation of the symptoms of the disease being treated.
The term "selective alpha la adrenergic receptor
antagonist," as used herein, refers to an alpha la antagonist compound
which is at least ten fold selective for the human alpha la adrenergic
receptor as compared to the human alpha lb, alpha ld, alpha 2a, alpha
2b and alpha 2c adrenergic receptors.
The present invention also provides pharmaceutical
compositions comprising one or more compounds of this invention in
association with a pharmaceutically acceptable carrier. Preferably
these compositions are in unit dosage forms such as tablets, pills,
capsules, powders, granules, sterile parenteral solutions or
suspensions, metered aerosol or liquid sprays, drops, ampoules, auto-
injector devices or suppositories; for oral, parenteral, intranasal,
sublingual or rectal administration, or for administration by inhalation
or insufflation. Alternatively, the compositions may be presented in a
form suitable for once-weekly or once-monthly administration; for
example, an insoluble salt of the active compound, such as the decanoate
salt, may be adapted to provide a depot preparation for intramuscular
injection. For preparing solid compositions such as tablets, the
principal active ingredient is mixed with a pharmaceutical carrier, e.g.
conventional tableting ingredients such as corn starch, lactose, sucrose,
sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or
gums, and other pharmaceutical diluents, e.g. water, to form a solid
preformulation composition containing a homogeneous mixture of a
compound of the present invention, or a pharmaceutically acceptable
salt thereof. When referring to these preformulation compositions as
homogeneous, it is meant that the active ingredient is dispersed evenly
throughout the composition so that the composition may be readily
subdivided into equally effective unit dosage forms such as tablets, pills
and capsules. This solid preformulation composition is then subdivided
into unit dosage forms of the type described above containing from 0.1 to
about 500 mg of the active ingredient of the present invention. The
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tablets or pills of the novel composition can be coated or otherwise
compounded to provide a dosage form affording the advantage of
prolonged action. For example, the tablet or pill can comprise an inner
dosage and an outer dosage component, the latter being in the form of an
envelope over the former. The two components can be separated by an
enteric layer which serves to resist disintegration in the stomach and
permits the inner component to pass intact into the duodenum or to be
delayed in release. A variety of materials can be used for such enteric
layers or coatings, such materials including a number of polymeric
acids and mixtures of polymeric acids with such materials as shellac,
cetyl alcohol and cellulose acetate.
As used herein, the term "composition" is intended to
encompass a product comprising the specified ingredients in the
specified amounts, as well as any product which results, directly or
indirectly, from combination of the specified ingredients in the specified
amounts.
The liquid forms in which the novel compositions of the
present invention may be incorporated for administration orally or by
injection include aqueous solutions, suitably flavoured syrups, aqueous
or oil suspensions, and flavoured emulsions with edible oils such as
cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and
similar pharmaceutical vehicles. Suitable dispersing or suspending
agents for aqueous suspensions include synthetic and natural gums
such as tragacanth, acacia, alginate, dextran, sodium
carboxymethylcellulose, methylcellulose, polyvinyl-pyrrolidone or
gelatin.
Where the processes for the preparation of the compounds
according to the invention give rise to mixtures of stereoisomers, these
isomers may be separated by conventional techniques such as
preparative chromatography. The compounds may be prepared in
racemic form, or individual enantiomers may be prepared either by
enantiospecific synthesis or by resolution. The compounds may, for.-
example, be resolved into their component enantiomers by standard
techniques, such as the formation of diastereomeric pairs by salt
formation with an optically active acid, such as (-)-di-p-toluoyl-d-tartaric
acid and/or (+)-di-p-toluoyl-1-tartaric acid followed by fractional
crystallization and regeneration of the free base. The compounds may
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also be resolved by formation of diastereomeric esters or amides,
followed by chromatographic separation and removal of the chiral
auxiliary. Alternatively, the compounds may be resolved using a chiral
HPLC column.
During any of the processes for preparation of the
compounds of the present invention, it may be necessary and/or
desirable to protect sensitive or reactive groups on any of the molecules
concerned. This may be achieved by means of conventional protecting
groups, such as those described in Protective Groups in Organic
Chemistry, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene &
P.G.M. Wuts, Protective Groups in Or nic Synthesis, John Wiley &
Sons, 1991. The protecting groups may be removed at a convenient
subsequent stage using methods known from the art.
The specificity of binding of compounds showing affinity for
the alpha la receptor is shown by comparing affinity to membranes
obtained from tranfected cell lines that express the alpha la receptor and
membranes from cell lines or tissues known to express other types of
alpha (e.g., alpha ld, alpha lb) or beta adrenergic receptors. Expression
of the cloned human alpha ld, alpha lb, and alpha la receptors and
comparison of their binding properties with known selective antagonists
provides a rational way for selection of compounds and discovery of new
compounds with predictable pharmacological activities. Antagonism by
these compounds of the human alpha la adrenergic receptor subtype
may be functionally demonstrated in anesthetized animals. These
compounds may be used to increase urine flow without exhibiting
hypotensive effects.
The ability of compounds of the present invention to
specifically bind to the alpha la receptor makes them useful for the
treatment of BPH. The specificity of binding of compounds showing
affinity for the alpha 1a receptor is compared against the binding
affinities to other types of alpha or beta adrenergic receptors. The
human alpha adrenergic receptor of the la subtype was recently - --
identified, cloned and expressed as described in PCT International
Application Publication Nos. W094/08040, published 14 April 1994 and
WO 94J21660, published 29 September 1994. The cloned human alpha la
receptor, when expressed in mammalian cell lines, is used to discover
ligands that bind to the receptor and alter its function. Expression of the
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cloned human alpha 1d, alpha lb, and alpha la receptors and
comparison of their binding properties with known selective antagonists
provides a rational way for selection of compounds and discovery of new
compounds with predictable pharmacological activities.
Compounds of this invention exhibiting human alpha 1a
adrenergic receptor antagonism may further be defined by
counterscreening. This is accomplished according to methods known in
the art using other receptors responsible for mediating diverse biological
functions. S a e. , PCT International Application Publication No.
W094/10989, published 26 May 1994; U.S. Patent No. 5,403,847, issued
April 4, 1995]. Compounds which are both selective amongst the various
human alphal adrenergic receptor subtypes and which have low affinity
for other receptors, such as the alpha2 adrenergic receptors, the 13-
adrenergic receptors, the muscarinic receptors, the serotonin receptors,
and others are particularly preferred. The absence of these non-specific
activities may be confirmed by using cloned and expressed receptors in
an analogous fashion to the method disclosed herein for identifying
compounds which have high affinity for the various human alphal
adrenergic receptors. Furthermore, functional biological tests are used
to confirm the effects of identified compounds as alpha la adrenergic
receptor antagonists.
The present invention also has the objective of providing
suitable topical, oral, systemic and parenteral pharmaceutical
formulations for use in the novel methods of treatment of the present
invention. The compositions containing compounds of this invention
as the active ingredient for use in the specific antagonism of human
alpha la adrenergic receptors can be administered in a wide variety
of therapeutic dosage forms in conventional vehicles for systemic
administration. For example, the compounds can be administered in
such oral dosage forms as tablets, capsules (each including timed
release and sustained release formulations), pills, powders,
granules, elixirs, tinctures, solutions, suspensions, syrups and w --
emulsions, or by injection. Likewise, they may also be administered
in intravenous (both bolus and infusion), intraperitoneal,
subcutaneous, topical with or without occlusion, or intramuscular
form, all using forms well known to those of ordinary skill in the
pharmaceutical arts. An effective but non-toxic amount of the
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compound desired can be employed as an alpha 1a antagonistic
agent.
Advantageously, compounds of the present invention
may be administered in a single daily dose, or the total daily dosage
may be administered in divided doses of two, three or four times
daily. Furthermore, compounds for the present invention can be
administered in intranasal form via topical use of suitable intranasal
vehicles, or via transdermal routes, using those forms of
transdermal skin patches well known to those of ordinary skill in
that art. To be administered in the form of a transdermal delivery
system, the dosage administration will, of course, be continuous
rather than intermittent throughout the dosage regimen.
The dosage regimen utilizing the compounds of the
present invention is selected in accordance with a variety of factors
including type, species, age, weight, sex and medical condition of the
patient; the severity of the condition to be treated; the route of
administration; the renal and hepatic function of the patient; and the
particular compound thereof employed. A physician or veterinarian
of ordinary skill can readily determine and prescribe the effective
amount of the drug required to prevent, counter or arrest the
progress of the condition. Optimal precision in achieving
concentration of drug within the range that yields efficacy without
toxicity requires a regimen based on the kinetics of the drug's
availability to target sites. This involves a consideration of the
distribution, equilibrium, and elimination of a drug.
In the methods of the present invention, the compounds
herein described in detail can form the active ingredient, and are
typically administered in admixture with suitable pharmaceutical
diluents, excipients or carriers (collectively referred to herein as
"carrier" materials) suitably selected with respect to the intended
form of administration, that is, oral tablets, capsules, elixirs, syrups
and the like, and consistent with conventional pharmaceutical - -
practices.
For instance, for oral administration in the form of a
tablet or capsule, the active drug component can be combined with an
oral, non-toxic pharmaceutically acceptable inert carrier such as
ethanol, glycerol, water and the like. Moreover, when desired or
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necessary, suitable binders, lubricants, disintegrating agents and
coloring agents can also be incorporated into the mixture. Suitable
binders include, without limitation, starch, gelatin, natural sugars
such as glucose or beta-lactose, corn sweeteners, natural and
synthetic gums such as acacia, tragacanth or sodium alginate,
carboxymethylcellulose, polyethylene glycol, waxes and the like.
Lubricants used in these dosage forms include, without limitation,
sodium oleate, sodium stearate, magnesium stearate, sodium
benzoate, sodium acetate, sodium chloride and the like.
Disintegrators include, without limitation, starch, methyl cellulose,
agar, bentonite, xanthan gum and the like.
The liquid forms in suitably flavored suspending or
dispersing agents such as the synthetic and natural gums, for example,
tragacanth, acacia, methyl-cellulose and the like. Other dispersing
agents which may be employed include glycerin and the like. For
parenteral administration, sterile suspensions and solutions are
desired. Isotonic preparations which generally contain suitable
preservatives are employed when intravenous administration is desired.
The compounds of the present invention can also be
administered in the form of liposome delivery systems, such as small
unilamellar vesicles, large unilamellar vesicles and multilamellar
vesicles. Liposomes can be formed from a variety of phospholipids,
such as cholesterol, stearylamine or phosphatidylcholines.
Compounds of the present invention may also be
delivered by the use of monoclonal antibodies as individual carriers to
which the compound molecules are coupled. The compounds of the
present invention may also be coupled with soluble polymers as
targetable drug carriers. Such polymers can include polyvinyl-
pyrrolidone, pyran copolymer, polyhydroxypropylmethacryl-
amidephenol, polyhydroxy-ethylaspartamidephenol, or polyethyl-
eneoxidepolylysine substituted with palmitoyl residues.
Furthermore, the compounds of the present invention may be coupled--
to a class of biodegradable polymers useful in achieving controlled
release of a drug, for example, polylactic acid, polyepsilon
caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals,
polydihydro-pyrans, polycyanoacrylates and cross-linked or
amphipathic block copolymers of hydrogels.
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Compounds of this invention may be administered in
any of the foregoing compositions and according to dosage regimens
established in the art whenever specific blockade of the human alpha
la adrenergic receptor is required.
The daily dosage of the products may be varied over a wide
range from 0.01 to 1,000 mg per adult human per day. For oral
administration, the compositions are preferably provided in the form of
tablets containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0
and
100 milligrams of the active ingredient for the symptomatic adjustment
of the dosage to the patient to be treated. A medicament typically
contains from about 0.01 mg to about 500 mg of the active ingredient,
preferably, from about 1 mg to about 100 mg of active ingredient. An
effective amount of the drug is ordinarily supplied at a dosage level of
from about 0.0002 mg/kg to about 20 mg/kg of body weight per day.
Preferably, the range is from about 0.001 to 10 mg/kg of body weight per
day, and especially from about 0.001 mg/kg to 7 mg/kg of body weight per
day. The compounds may be administered on a regimen of 1 to 4 times
per day.
Compounds of this patent disclosure may be used alone at
appropriate dosages defined by routine testing in order to obtain optimal
antagonism of the human alpha la adrenergic receptor while
minimizing any potential toxicity. In addition, co-administration or
sequential administration of other agents which alleviate the effects of
BPH is desirable. Thus, in one embodiment, this includes
administration of compounds of this invention and a human
testosterone 5-a reductase inhibitor. Included with this embodiment are
inhibitors of 5-alpha reductase isoenzyme 2. Many such compounds are
now well known in the art and include such compounds as PROSCAR~,
(also known as finasteride, a 4-Aza-steroid; see US Patents 4,377,584 and
4,760,071, for example). In addition to PROSCAR~, which is principally
active in prostatic tissue due to its selectivity for human 5-a reductase
isozyme 2, combinations of compounds which are specifically active in-
inhibiting testosterone 5-alpha reductase isozyme 1 and compounds
which act as dual inhibitors of both isozymes 1 and 2, are useful in
combination with compounds of this invention. Compounds that are
active as 5a-reductase inhibitors have been described in W093/23420, EP
0572166; WO 93/23050; W093/23038, ; W093/23048; W093/23041;
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W093/23040; W093/23039; W093/23376; W093/23419, EP 0572165;
W093/2305I.
The dosages of the alpha la adrenergic receptor and
testosterone 5-alpha reductase inhibitors are adjusted when
combined to achieve desired effects. As those skilled in the art will
appreciate, dosages of the 5-alpha reductase inhibitor and the alpha
1a adrenergic receptor antagonist may be independently optimized
and combined to achieve a synergistic result wherein the pathology is
reduced more than it would be if either agent were used alone. In
accordance with the method of the present invention, the individual
components of the combination can be administered separately at
different times during the course of therapy or concurrently in
divided or single combination forms. The instant invention is
therefore to be understood as embracing all such regimes of
simultaneous or alternating treatment and the term "administering"
is to be interpreted accordingly.
Thus, in one preferred embodiment of the present
invention, a method of treating BPH is provided which comprises
administering to a subject in need of treatment any of the compounds
of the present invention in combination with finasteride effective to
treat BPH. The dosage of finasteride administered to the subject is
about 0.01 mg per subject per day to about 50 mg per subject per day in
combination with an alpha 1a antagonist. Preferably, the dosage of
finasteride in the combination is about 0.2 mg per subject per day to
about 10 mg per subject per day, more preferably, about 1 to about 7
mg per subject to day, most preferably, about 5 mg per subject per
day.
For the treatment of benign prostatic hyperplasia,
compounds of this invention exhibiting alpha la adrenergic receptor
blockade can be combined with a therapeutically effective amount of a
5a-reductase 2 inhibitor, such as finasteride, in addition to a 5a-
reductase 1 inhibitor, such as 4,713-dimethyl-4-aza-5a-cholestan-3- - --
one, in a single oral, systemic, or parenteral pharmaceutical dosage
formulation. Alternatively, a combined therapy can be employed
wherein the alpha la adrenergic receptor antagonist and the 5a-
reductase 1 or 2 inhibitor are administered in separate oral,
systemic, or parenteral dosage formulations. See, e.g., U.S. Patent
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No.'s 4,377,584 and 4,760,071 which describe dosages and
formulations for 5a-reductase inhibitors.
Abbreviations used in the instant specification, particularly
the Schemes and Examples, are as follows:
BCE = bromochloroethane
Boc or BOC = t-butyloxycarbonyl
BOPCl = bis(2-oxo-3-oxazolidinyl)phosphinic chloride
Cbz-Cl or CBZCI= benzyloxycarbonyl chloride
CSA = 10-Camphorsulfonic acid
DAST = diethylaminosulfurtrifluoride
DEAD = diethylazodicarboxylate
DMF = N,N-dimethylformamide
DMSO = dimethylsulfoxide
EDCI = 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride
Et = ethyl
Et3N = triethyiamine
EtOAc = ethyl acetate
EtOH = ethanol
FABLRMS = fast atom bombardment low resolution mass
spectroscopy
HMPA = hexamethylphosporamide
HPLC = high performance liquid chromatography
HOAc = acetic acid
HOBt = 1-hydroxy benzotriazole hydrate
i-PrOH = 2-propanol
i-Pr2NEt = diisopropylethylamine
LAH = lithium aluminum hydride
LDA = lithium diisopropyl amide
mCPBA = meta-chloroperbenzoic acid
Me = methyl . -
MeOH = methanol
NMR = nuclear magnetic resonance
PCTLC = preparative centrifugal thin Iayer
chromatography
PEI = polyethylenimine
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Ph = phenyl
pTSOH = p-toluenesulfonic acid
RT = retention time
TEBAC = benzyltriethylatnmonium chloride
r~ TFA = trifluoroacetic acid
THF = tetrahydrofuran
TLC = thin layer chromatography
TMS = trimethylsilyl
TMSCN = trimethylsilylcyanide
The compounds of the present invention can be prepared
readily according to the following reaction schemes and examples, or
modifications thereof, using readily available starting materials,
reagents and conventional synthesis procedures. In these reactions, it
is also possible to make use of variants which are themselves known to
those of ordinary skill in this art, but are not mentioned in greater detail.
Unless otherwise indicated, all variables are as defined above.
Many of the compounds claimed within this invention can
be assembled as outlined in a general fashion in Scheme 1. Reductive
amination of a hydroxy amino derivative with a cycloalkanone provides
an intermediate which can be carried on via either of the tyvo outlined
methods; conversion of the hydroxy group to a halide or tosylate and
displacement with an anionic equivalent of one of the Q groups.
Alternatively, the hydroxy group can be manipulated into an al~aino
group and acylated or alkylated to supply the desired amide as urea
linked analogs.
An example of this approach is highlighted where the
central portion of the antagonist is assembled from N-protected 3-
hydroxy azetidine. Conversion of the hydroxy to a protected amino group
and deprotection of the azetidine nitrogen provided the appropriate
amine for reductive amination with a cycloalkanone, which after
deprotection was acylated with an activated Q species. This was
accomplished with the 3-aminomethyl azetidine linker in an analogous
fashion.
Derivatives where the central molecular framework is a
4-amino piperidine were prepared from isonipecotic acid via the
sequence outlined in Scheme 2. Isonipecotic acid was converted to 4-
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(tert-butoxycarbonyl)amino piperidine in three chemical steps.
Reductive amination with 4-cyano 4-phenyl cyclohexanone provided a
separable mixture of cis and trans products, which after Boc
deprotection and acylation provided the desired antagonists.
The 3-amino piperidinyl bridged compounds were
assembled in analogous fashion starting from nipecotic acid, Scheme 3.
The corresponding 3-amino pyrroldinyl analogs were obtained by
substituting the commercially available 3-(tent-butoxycaronyl)amino
pyrrolidine for the corresponding piperidinyl material as shown in
Scheme 4.
Selective acylation of the primary amines was accomplished
by treatment of the amines with nearly equimolar quantities of the
activated termini species (i.e., the "Q" groups). The activated termini
species comprising the "Q" groups are readily prepared by one of
ordinary skill in the art. For example, unsubstituted, alkyl- and
cycloalkyl-substituted oxazolidinones are prepared and activated in
general by published and well developed chemistry, in particular, of
Evans. [Evans, D.A.; Nelson, J.V.; Taber, T.R. Top. Stereochem. 13, 1
(1982)] The starting materials, in general, are natural and unnatural
amino acids. For instance, some of the preferred compounds are
prepared from substituted phenyl glycine derivatives, which after
reduction of the carboxylate and a phosgene equivalent mediated
cyclization provides the substituted oxazolidinone ring system.
Deprotonation with n-butyl lithium and addition to a THF solution of p-
nitrophenylchloroformate produces the stable, isolable
"activated"oxazolidinone (oxa).
Oxazolidinones substituted with carboxylate, carboxamide,
and hydroxymethyl are prepared by hydroxyamination of olefins to
provide protected aminoalcohols, using procedures as described in
Sharpless et al., Angew. Chem. Int. Ed. Engl., 35, 2813 (1996).
Deprotection under standard conditions followed by a phosgene
equivalent to mediate cyclization provides the substituted oxazolidinone
ring system. Deprotonation with a strong base, for example, lithium
bis(trimethylsilyl)amide, and addition to a THF solution of p-
nitrophenylchloroformate produces the stable, isolatable "activated"
oxazolidinone.
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Dihydropyrimidinones are prepared by condensation
reaction of the aldehyde, urea and a 1,3-acetoacetate type derivative
catalyzed by a Lewis Acid, a copper (I) species and acetic acid.
Activation was accomplished by treatment with a strong base, for
instance, LiN(TMS)2, followed by addition to a THF solution of p-
nitrophenylchloroformate.
Hydantoins and cycloimide were prepared in two chemical
steps from ketones as outlined in the literature. More specifically,
hydantoins were prepared according to known methodology, e.g., J.J.
Edmunds et al., J. Med. Chem. 1995, 38, pp. 3759-3771; J.H. Poupart et
al., J. Chem. Res. 1979, pp. 174-175. Saccharins were prepared
according to known methods, e.g., page 40 and Examples 21 and 22 of
PCT International Application Publication No. W096/25934, published
August 29, 1996.
The dihydropyrimidinones and the unsubstituted, alkyl-
and cycloalkyl-substituted oxazolidinones were synthesized
independently in racemic form, and then separated utilizing preparative
chiral HPLC. Their optical rotations were recorded. Then they were
activated and reacted with prerequisite amines. From the receptor
binding studies, a preferred isomer was identified, the (+) rotational
isomer in each case. The absolute configurations were determined to be
(S) for both the dihydropyrimidinones and oxazolidinones by correlating
their optical rotations with x-ray crystal structures obtained of
fragments involved in the production of the antagonists.
The oxazolidinones substituted with carboxylate,
carboxamide, and hydroxymethyl were prepared in enantiomer-
enriched form and the assignments of (4S,5R) were made in accordance
with Sharpless et al., Angew. Chem. Int. Ed. Engl., 35, 2813 (1996)
Antagonists with cycloalkyl linking chains can be
assembled by reductive amination of the prerequisite amino alcohol and
a ketone, for example, N-{2-cyanophenyl)piperidin-4-one, Scheme 5.
Conversion of the hydroxy to a tosylate with tosyl anhydride, followed by
displacement by the sodium or lithium salt of the desired Q group
completes the synthesis of the targeted antagonists.
Some of the required ketones were readily assembled
following the sequence outlined in Scheme 6. For example, a substituted
benzyl nitrile, sulphone, etc. could be added to methyl acrylate (or other
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substituted acrylates), submitted to Dieckman cyclization, hydrolyzed
and decarboxylated providing appropriately substituted ketones.
Further modifications of the ketones can be accomplished following the
Dieckman cyclization, which provides the (3-keto ester which can be
either: (a) submitted to a reductive amination and carried on to final
product, (b) enolized and alkylated then reductively aminated,
deprotected and further manipulated providing further substituted
analogs; or (c) hydrolyzed and decarboxylated and run through the
above described conditions producing the desired antagonists.
Another strategy for the synthesis of some geminally
disubstituted cyclic ketones, in particular, 4,4-disubstituted
cyclohexanones was accomplished as outlined in Schemes 7A and 7B
starting from benzophenone derivatives and substituted methyl vinyl
ketones which under basic conditions lead to the 4,4-aryl cyclohex-2-en-1-
ones in good yield. Subsequent hydrogenation, reductive amination and
deprotection provided the appropriate acylation/alkylation precursors.
Alternatively, the 4,4-diaryl cyclohex-2-en-1-ones could be subjected to
Michael addition of selected nucleophiles, alkylation or aldolyzation of
the enolate of the resulting ketone then reductively aminated and carried
through the standard chemical transformation to provide further
elaborated antagonists.
The synthesis of some additional compounds of the present
invention is described in Schemes 8-17. Schemes 16-17 describe the
synthesis of the 3-aminomethyl-3-hydroxyazetidine and the 4-amino-3-
hydroxypyrrolidine intermediates. The 3-aminomethyl-3-
hydroxyazetidine was assembled from the commercially available N-
protected 3-hydroxyl azetidine as outlined Scheme 16. Swern oxidation of
the alcohol with dimethylsulfoxide and oxalyl chloride provided the
azetidinone. The zinc iodide catalyzed addition of TMSCN produced the
cyanohydrin. Subsequent LAH reduction of the nitrile and two
protecting group manipulations yielded the key intermediate required
for the reductive aminations with the cyclohexanones. Deprotection-o~
the N-BOC carbamate and acyclation with preferred activated "Q"-
groups furnished the final targets. The synthesis of the 4-amino-3-
hydroxypyrrolidine intermediate began with 3,4-pyrroline. BOC
protection of the amine followed by mCPBA oxidation provided the
epoxidation. Subsequent sodium azide opening of the epoxide and
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triphenylphosphine/water mediated reduction produced 4-amino-N-1-
(1,1-dimethylethoxycarbonyl)-3-hydroxypyrrolidine. After two protecting
manipulations the key amino intermediate was alkylated by reductive
amination reactions with cyclohexanones. Following the cleavage of the
CBZ protecting group acyclation with preferred activated "Q"-groups
furnished the final targets.
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Scheme 1
2R m
~'' OH
CN n HN~) ( ~ CN n
/i
(R20)r p (R20)r 2 R3 m
o ~ ~~O AcOH/MeOH o ~ ~N OH
A NaCNeH3
i P
CBr4
PPh3
1 ) Tos20
CN 2) NaN3
(R2o)/~ n 2 3} PPh3/H20
r o ~ ~ N R m q,
~'Br LG Q'
P (R EDCI, HOBT
H-Q
CN i R2 R m O
/ n
R .., , ,
20 / ~ N
( )r (~~ R2 R m ~ ~ H Q'
o N Q p
p l_G = leaving group, e.g.,
\ w (X)S
\ / N02
i i
wN C02Me
Q' = w
O N I OMe N
H ~-O
O
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Scheme 1 (cont'd)
~ CN
/ n
CN n AcOHIMeOH
NaCNBH3 (R2o)
_( ~.. .. o~ N-1
A "
HCI
t ) Tos20 I ~ CN n EtOAc
2) NaN3 zo ~ ~
3) PPh3lH20 (R )r ~ ~,
4 8oc20 0~ ~N
5)H2/Pd-C ~--NH2
Ph
--N~OH
O
Ph I ~ CN
I.G~Q'
n
(R2o) /
o~ N ~ D,
~- N
H
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Scheme 1 (cont'd)
CN'
CN n AcOH/MeOH 20 ~~ / n
(R2o) ~ NaCNBH3 (R )r
o~ O o~ N
HN\~ NHBOC
A NHBOC
1 ) LAH HCI
2) Boc20 EtOAc
3 H2/Pd-C
Ph
-f~- CN
Ph I CN
/ n
(Rzo)
o~ N
NH2
O
CN
n LG~Q
(R2o)
o~ N H
N ~Q
'IO
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SCHEME 2
Ti(OiPr)4 \ CN
\ NaCNBH3 I / . HC~
I CN MeOH EtOAc
N
O HN I \ NHBOC
N NHBOC / ,CN
1 ) CBZCI
2) DPPA ~ ~~~'N
C02H Et3N/tBuOH NHBOC
3) H2/Pd-C
I \
CN
\ ~ ~ F
CN
/ . F
N /I
N N \
1 ) 3,4-F2PhC02H H
NH EDCI
HOBt
\ 2) HCI/EtOAc
'CN
,N CN
/ . F
NH2 'N O / F
N
H
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SCHEME 3
Ti(OiPr)4 CN
\ NaCNBH3 I / ,
MeOH
/ CN NHBOC
N
O HN NHBOC \ HCI ,-
/ ' CN EtOAc
-NH 1) CBZCI
2) DPPA ~ ~.,N NHBOC
C02H Et3N/tBuOH
3) H2/Pd-C
F
CN \ I C02Me
\ / . F ~ Oi
CN N N NH
/ . N
N NH2 O O
CN
/ .
..,,N NH2
F /
C02Me
CN
F I / . F ~ \ i
F -O
N N H
N
O / O O
'I C02Me 4
15 = O~N
1 , o H 1
02N °~ _.. __
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Scheme 3 (cont'd)
I CN
HO \ F
N NH2 O
F
\ EDCI
I CN HOBt
/ .
~,,N NH2 I CN
/ .
F H
.,. N N
F \ ~ \ F
\ O
16 O O I CN / F
/ . 5
02N ~ ~ O O H
N N
F \ F
\ O
I CN F \ ~ 6 F
/ , H
N N~ O
IOI O
7
F
I / ' CN F
N N O
N
O O
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SCHEME 4
Ti(OiPr)4 CN
\ NaCNBH3 I / .
CN MeOH
/ NHBOC
N
O HN NHBOC \ HCI _
C N EtOAc
/ .
-,,N NHBOC
\ F / C02Me
CN
\ i
\ F \ O
/ . CN N N NH
N~ ~ ~
N NH2 O O
9
/ ' CN 15
.,,.N NH2
F
\ / C02Me
CN
/ . F H \ O~
N~N~ NH
IOI I IO
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SCHEME 4 (Cont'd)
CN
/ .
Ris F
NH2 HO
CN N~ ~ ~ F R'3 = H or iPr
O
EDCI, HOBt
-,, N N H2 \
CN
/ . R~s F
H
N
16 N ' / F
\ 11 O
CN
/ . Rts
\ F / .,. N N \ F
/ 'CN \
F N N O 10 ~F
N
O O
13
F
'CN \
/ F
H O
~.,N N~N
IOI ~O
14
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SCHEME 5
AcOH or
Ti(OiPr)4 ~ CN
\ NaCNBH3 /
CN MeOH 1-2
1-2 N (CH2)o-20H
p HN (CH2)o-20H
0-2
F ~ 0-2
Tos20
F I\
CN
/ /
1-2
HN N (CH2)o_20Tos
nBuLi
O 02
O F
/ CN F \ I
1-2 ~
N N 'O
0-2 O
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SCHEME 5 (cont'd)
R13 ~ CN
R14 / R1g R14 O
H~Y ~ 1 _2
O N ~ i0-2 Y NRa
Z N
R8 0_2 Z
R13 \
R8 R14 NaH ~ CN Ra
O ~ / O N R1s
O~ i_2
[~ ~ N R 14
O (X)s ~ 0_2 O
HN ~ ~ \
O.;S1 / ~ / CN
O i _2 O (X)s
N 0-2 ~ N
0-2 O,.SO
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SCHEME 6
For example,
~C02Me ( ~ R KOtBu
~~R ~ / C02Me
(R2~)~ Triton-B (R2~)~ THF
'--~--J tBuOH
or heteroaryl or heteroaryl
C02Me
R = CN, S02alk, others
w ( w R
R C02Me AcOH ! /
R2' (R21)~
Aq HCI ~ O
or heteroaryl O or heteroaryl A
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SCHEME 7A
~S~
\ O O
/ O ~ H2/Pd-C
Base
I
O
HN~ NHP
NHBOC
AcOH/MeOH
NaCNBH3
P = BOC HCI
P = H~HCI ~ EtOAc
O , F
F N~ \ I
O ~ I EDCI \\// ~\ H F
HO F HOBt
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SCHEME 7B
~S~
I \ O
/ O ~ H2/Pd-C
Base
I
H
O H2N~ NHBOC N ~ NHP
AcOH/MeOH
NaCNBH3
P = BOC HCI
EtOAc
P = H~HCI
O / F
F N I
O ~ I EDCI ~ H \ F
HO F HOBt
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Scheme 8
LDA, PhN SO CF O
p ( 2 3)2
O F3C.S~ O
O
(R21)
Zn I
R2' = H, CN, C02Et, F, CF3, OMe, OR6 ~ ~ THF
Pd[P(Ph)3Ja
(RZi )~ O
(R2~ H2, Pd/C
EtOAc
1 N HCI 1. ~NHBOC
Dioxane HN
O NaCNBH3 2~ N~ NH3C1
(R21)r THF, MeOH (RJ )r
2. HCI(g), EtOAc
/ /
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SCHEME 8 (Cont'd)
N~NH3C1
(R11)r
O
02N ~ ~ O~Q, O
Q,
N~ N
TEA, DMF
(R2~) \
(X)s (X)s
I~\
/ /
wN C02Me ~N
Q -
O N OMe ~-O
H O
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Scheme 9
0 0
LDA, PhN(S02CF3) O
O 'S.O
O THF, -78°C F3C ~ '~
O
(R21)
Znl
R21 = H, CN, C02Et, F, CF3, OMe, OR6 ~ ~ THF
N Pd[P{Ph)sJa
O
{Rz~ )r O
(R2~)
r 'O H2, Pd/C I ~~ a
a - iN
~ N EtOAc
1 N HCI
Dioxane
{R21) O 1HN~NHBOC
r
N~ NH3C1
NaCNBH3 (R )r
I THF, MeOH
~N
2. HCI(g), EtOAc ~ N
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SCHEME 9 (Cont'd)
N~ NH3C!
(R21)r
TEA, DMF O
N OII N N ~Q'
i
02N ~ ~ O ~Q, H
(R2t)r ~N
(X)s (X)s
\\
~N C02Me
_ ! N
O ~ OMe ~-O
O H O
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Scheme 10
R21 ~R21)r O
1. n-BuLi, THF, -78°C I O
/ 2. O OH
~O
R21 = H, CN, Me, OMe,
CF , F, OR6 O 1 N HCI
s Dioxane
O
~R2~ )r
~R21)r O H2, Pd/C ~ \ /
\ EtOAc /
/
1' ~-NHBOC
HN
NaCNBH3
THF, MeOH O
2. HCI(g), EtOAc O
\ O O
N~'' NH3C1 O N /
21 z N~ N
~R ~ )r H
TEA, DMF
v
/ ~X)s (X)S ~R2~)r I /
i ~/
wN C02Me ~N
Q =
O N OMe ~-O
H O
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Scheme 11 O
\ B~ 1. n-BuLi, THF, -78°C ~ ~ p
O _
2. N OH
O 1 N HCI
O Dioxane
O
1 HN~ NHBOC \ pH
,N
NaCNBH3
THF, MeOH
N~ NHBoc
NH3C1
\ HCi(g)
~ N O EtOAc
O TEA, DMF
02N ~ ~ O~Q,
O
(X)S
(X)s N~' N Q'
\ H
w C02Me ~ ~ N OH
w
N I OMe N
O H ~-O
O
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Scheme 12
(R2i )
r
\ Br 1. n-BuLi, THF, -78°C I O
O
OH
2.
(R21) ~O
r ~ 1 N HCI
O Dioxane
O
(R2~
1' ~NHBOC I \ OH
H N ~~// /
NaCNBH3
THF, MeOH
N~ NH3C1
N~ NHBoc 21
(R
HCI(g)
OH
EtOAc
(R2t )r O
II TEA, DMF
Q2N ~ ~ O~Q,
O
(X)S
(X)s N~ N Q'
\ R2rt H
/ ~ ~ )r
/ \\
wN C02Me ~ / OH
p ~ I OMe \N
O H ~-O
O
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Scheme 13
HO OH O
Br 1. n-BuLi, THF, -78°C ~ I ~ O
O
OH
R21 O R21
O K2CO3
Rz1 = H, CN, Me, OMe, BnBr
CF3, F, OR6 DMF
/ ~ O
O 1 N HCI
Dioxane ~ _
O ~ / '~ ~ \ OH
/
R2i
R2~
H2, Pd/C
EtOAc
O
HO
a
1HN~NHBOC N NH CI
c,
NaCNBH3 HO
v
R2' THF, MeOH
2. HCI(g), EtOAc
R2t
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Scheme 13 (cont'd)
HC
TEA, DMF
O O
Ra ~Q.
N~ N Q'
HO H
v
(X)S
/ ~ ~~ R2~
w C02Me
N
q' O~ N I OMe \N
/~-O
O
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Scheme 14
h Iw I \ \
O O
N ZnBr N / H2, Pd (10%)
/ Pd(PPh THF ~ EtOH O
3)4~ ,
O O
OTf
\ \ 1. HN~ NHBoc
mCPBA ~ ~ HCI, HOAc, H20 ~ ~ NaCN~BH3, MeOH
-~. O ~ N ~~~ O
CHC13 ~ 2. chromatography
O O
\ \
HCI(g); Na2C03
N
~,,NHBoc
~~' N ~ ~. ,.~ NH2
O
F
F
F
02N ~ ~ O~N ~ C02Me I \ ~ F
Me
H II
DMF ''~~~"'N~N C02Me
O
Me
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Scheme 15
0
0
O BnOCONH2 NH O
O H MeOH I Me ~~OCI, NaOH I Me
HZ~ F a H
(DHQ)ZPHAL F
KzOs02(OH)2
F
NHz O
H2. Pd/C LiHMDS
O H Me tnph~ .~02Me -~
F ~pr2~t H p-NOZPhOCOCI
O
R z~ F
OZN ~1 Rz~ F
NHz I
02Me
'Pr?]vlEt "' O Y~OzMe
O
F F
R
R F I CHCl3/NH3 z' F
",t'1 O ~02Me mN O ~~ONHz
~N~ silica gcl
H
O O
10
20
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Scheme 15 (cont'd)
F F
F
F F
LiBH4 2.3 dihydropy~an
~,C 02M e ----i .O H
CSA, CHZCIZ
H ~ H N
a~~'~'O
F
F
R~
LiHMDS 02N O~ ~mN~
N H2
p-NO~PhOCOCI '~
' Pr.,NEt
F F
F
R2~ F
pTsOH ~..~ O
mN~ ~ ~,,-O H
MeOH ~ N~l
~''O
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SCHEME 16
Swern ~ TMSCN
Ph N Oxidatn Ph N Znl2
a
OH O
Ph LAH ~ BOC20
Ph' 'N AIC13 Ph N NH2
''' I'CN
OH OH
Ph
H2/Pd-C
Ph~N -' HN
~NHBOC EtOH ~NHBOC
'~O~ H~' OH
+ ~ / NaCNBH3
CN ~~ AcOH, MeOH
O
1 ) HCI-EtOAc Final
2) Activated Products
(+)-DHP
or
OH (S)-OXA
CN ~N
NHBOC
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SCHEME 17
HN BOC20 gpCN ~ mCPBA
NaHC03
BoCN Na~ BoCN -off PP~
p DMF H20
N3
BOCN _pH CBZCI BOCN
-pH HCI
NH2 NHCBZ EtOAC
HN _pH + ~ / NaCNBH3
CN ~~ AcOH, MeOH
NHCBZ p
\ ~ 1 ) H2/Pd-C Final
CN ~N 2) Activated Products
-OH (+)-DHP
or
NHCBZ {S)-OXA
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The following examples are provided to further define the
invention without, however, limiting the invention to the particulars of
these examples.
Examples 1-12 were prepared according to Schemes 8 and 9.
EXAMPLE 1
(4S)-3-(1-[4-(2-Cyano-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl5
4-(3,4-difluoro-phenyl)-6-methyoxymethyl-2-oxo-1,2,3,4-tetra
hydropyrimidine-5-carboxylic acid methyl ester hydrochloride
F
CN
N O ~ ~ F
"", //
N
H N
O~ ~ C02Me
N
H OMe . HCl
Step A: Trifluoromethanesulfonic acid 1,4-dioxa-spiro[4.5]dec-7-en-
8-yl ester
To a solution of diisopropylamine (7.6 mL, 54.3 mmol) in 200
mL THF cooled to -78°C was added n-butyllithium (21.8 mL 2.5 M in
hexane, 54.3 mmol) under argon.The solution was stirred for 10
minutes, then a solution of 1,4-cyclohexanedione monoethylene ketal (8.5
g, 54.3 mmol) in 75 mL THF was added slowly. The solution was stirred
for 10 min, then a solution of N-phenyltrifluoromethane sulfonamide
(19.5 g, 54.3 mmol) in 150 mL THF was added slowly. The reaction
solution was warmed to r.t., poured onto saturated sodium bicarbonate,
and extracted with ethyl acetate. The combined organic layers were
washed with saturated sodium chloride, dried with magnesium sulfate,
and concentrated in vacuo. The crude material was passed through ~ -
silica (3% methanol, dichloromethane) to give the product as an oil.
1H NMR 8g (CDC13) 5.7-5.6 (m, 1H), 4.1-4.0 (m, 4H), 2.6-2.5
(m, 2H), 2.5-2.4 (m, 2H), 1.9 (t, 2H, J 6.6).
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Step B: 2-(1,4-Dioxaspiro[4.5]dec-7-en-8-yl)benzonitrile
To a solution of trifluoromethanesulfonic acid 1,4-dioxa-
spiro[4.5]dec-7-en-8-yl ester (12.5 g, 45.2 mmol) in THF (100 mL) at room
temperature was added a solution of iodozinc benzonitrile ( 100 mL of a
0.5 N solution in THF, 50 mmol) and palladium
tetrakistriphenylphophine (1 g, 0.8 mmol). The reaction was heated to
80°C for 1 hour. The reaction was cooled to room temperature and
poured into saturated sodium bicarbonate ( 1L), and extracted with ethyl
10 acetate. The combined organic layers were washed with saturated
sodium chloride, dried with magnesium sulfate, and concentrated in
vacuo. The crude material was chromatographed over silica gel eluting
with 25% ethylacetate/hexane to give the product as a pale yellow oil.
1H NMR, &f~ (CDC13) 7.7-7.6 (m, 1H), 7.52 (dt, 1H, J = 2, 7
Hz),7.4-7.26 (m, 2H), 5.95-5.88 (m, IH), 4.05-4.0 (m, 4H), 2.7-2.6 (m, 2H),
2.55-2.48 (m, 2H), 1.95 (t, 2H, J = 6.6 Hz).
Step C: 2-{1,4-Dioxaspiro[4.5]dec -8-yI)benzonitrile
20 To a suspension of 10% palladium on carbon ( 1.5g) in ethyl
acetate {200mL) at room temperature under argon was added a solutioin
of 2-(1,4-Dioxaspiro[4.5]dec-7-en-8-yl)benzonitrile (8.9 g, 36.7 mmol) in
ethyl acetate (50 mL). Hydrogen gas was then bubbled into the reaction
mixture until all the starting material was consumed. The reaction was
25 filtered through celite to remove the catalyst and the solution
concentrated to give the product as a colorless oil.
1H NMR, &f.~ (CDCl3) 7.65-7.6 (m, 1H), 7.54 (dt, 1H, J = 2, 7
Hz),7.46 (br d, 2H, J = 7 Hz), 7.28 (dt, 1H, J = 2, 7 Hz), 4.05-4.0 (m, 4H),
3.1-
3.0 (m, 1H), 2.0-1.7 (m, 8H)
Step D: 2-(4-Oxocyclohexyl)benzonitrile
To a stirring solution of 2-(1,4-Dioxaspiro[4.5]dec -8-
yl)benzonitrile (8.7 g, 35.7 mmol) in dioxane {200 mL) was added 1N HCl
(200 mL) and the reaction heated to 80°C for 2 hours. The reaction was
cooled to room temperature and poured into water (1 L) the mixture was
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extracted with ethyl acetate (3 X 250 mL). The combined organics were
washed with brine (200 mL) and then dried over anhydrous magnesium
sulfate, filtered and concentrated at reduced pressure to give the
product.
1H NMR 8H (CDC13) 7.65-7.6 (br d, 1H, J = 7 Hz), 7.59 (br t,
1H, J = 7 Hz),7.40-7.30 (m, 2H), 3.5 (tt, 1H, J = 3, 10 Hz), 2.7-2.5 (m, 4H),
2.35-2.2 (m, 2H), ), 2.05-1.90 (m, 2H)
Step E: cis and tracns 2-[4-(3R-3-Amino-pyrrolidin-1-yl)-cyclohexyl]-
benzonitrile dihydrochloride
To a solution of 2-(4-Oxocyclohexyl)benzonitrile (0.921 g, 4.63
mmol) and acetic acid (1.0 mL 17.5 mmol) in 30 mL methanol was
addded (3R)-(+)-3-(tert-butoxycarbonylamino)-pyrrolidine (1.298, 6.95
mmol) under argon. The solution was stirred for 2h, then sodium
cyanoborohydride (6.95 mL 1M in THF, 6.95 mmol) was added dropwise.
The solution was poured onto saturated sodium bicarbonate, and
extracted with ethylacetate. The combined organic layers were washed
with saturated sodium chloride, dried with magnesium sulfate, and
concentrated in vacuo. The crude material was passed through silica
(5~o methanol, ethylactate) to give 2:1 trans:cis 2-[4-(3-
tertbutoxycarbonylamino-pyrrolidin-1-yl)-cyclohexyl]-benzonitrile. The
trc~ns-2-[4-(3-tertbutoxycarbonylamino-pyrrolidin-1-yl)-cyclohexyl]-
benzonitrile (1.1 g, 2.98mmol) was dissolved in 100 mL ethylacetate, and
HCl gas was bubbled through the solution for 30 min. The solution was
concentrated in vacuo to give the tracns product.
1H NMR &H (CDC13) 7.75-7.60 (m, 2H), 7.55-7.45 (m, 1H),
7.40-7.35 (m, 1H), 4.20-3.20 (m, 7H), 3.15-3.00 (m, 2H), 2.80-2.50 (m, 1H),
2.40-2.00 (m, 5H), 1.90-1.60 (m, 4H).
The cis-2- [4- (3-tert -butoxycarbonylamino-pyrrolidin-1-yl) -
cyclohexyl]- benzonitrile (0.60 g, 1.6 mmol) was dissolved in 100 mL
ethylacetate, and HCl gas was bubbled through the solution for 30 min.
The solution was concentrated in vacuo, swished with hot ethyl acetate,
and filtered to give the cis product
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1H NMR ~ (CDCl3) 8.00-7.90 (m, 1H), 7.70-7.60 (m, 2H),
3.40-3.30 (m, 1H), 4.30-3.80 (m, 6H), 3.70-3.I0 (m, 3H), 2.85-2.70 (m, 1H),
2.55-1.75 (m, 9H).
Step F: 3-{1-[4-(2-Cyano-phenyl)-cyclohexyl]-3R-pyrrolidin-3-
ylcarbamoyl]-4-(3,4-difluoro-phenyl)-6-methoxymethyl-2-
oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl
ester hydrochloride
To a solution of trans-2-[4-(3R-3-Amino-pyrrolidin-1-yl)-
cyclohexyl]-benzonitrile dihydrochloride (300 mg, 0.880 mmol) and
triethylamine (0.37 mL, 2.64 mmol) in 3.0 mL DMF was added (+)-3-(4-
nitrophenoxycarbonyl)-4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester (462 mg,
0.968 mmol) under argon. The solution was stirred for 2 hour, poured
onto saturated sodium bicarbonate, and extracted with ethylacetate. The
combined organic layers were washed with saturated sodium chloride,
dried with magnesium sulfate, and concentrated in vacuo. The crude
material was passed through silica (3°l° methanol,
dichloromethane)
and washed with sodium carbonate. To a solution of the free base in
ethylacetate was added excess hydrogen chloride (1M in diethylether).
This solution was concentrated in vacuo to give the product.
Anal. Calcd. for Cg2H35F2N505~HC1: C, 59.67; H, 5.63; N,
10.87. Found: C, 60.10; H, 5.13; N, 11.16%.
The following compounds were prepared by procedures
substantially as described above for step F from either the cis or trans
product of step E. In the case of Examples 3 and 4, the (+)-(S)-3-(4-
nitrophenoxycarbonyl)-4-(3,4-difluorophenyl)-oxazolidin-2-one was
substituted for the (+)-3-(4-nitrophenoxycarbonyl)-4-(3,4-difluorophenyl)-
6-methoxymethyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylic acid
methyl ester
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EXAMPLE 2
(4S)-cis-3-{ 1-[4-(2-Cyano-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3
ylcarbamoyl}-4-(3,4-difluoro-phenyl)-6-methyoxymethyl-2-oxo-1,2,3,4
tetrahydropyrimidine-5-carboxylic acid methyl ester hydrochloride
F
CN / ~ F
N~ ~
N N
C02Ma
N
H OMe ~ HC1
Analysis: Calcd. for Cg2Hg5F2N5~5 ~ HCl ~ 0.4 H20
C, 59.01; H, 5.70; N,10.75
Found: C, 59.04; H, 5.65; N,10.47
EXAMPLE 3
trans-4S-4-(3,4-Difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{ 1-[4-
(2-Cyano-phenyl)-cyclohexyl]-3R-pyrrolidin-3-yl}amide hydrochloride
F
CN
~ "~~~ N O / \ F
N
H N
O~O ~ HCl
Analysis: Calcd. for C2~H2gF2N4Og ~ HCl ~ 0.2 H20 ~ 0.25 EtOAc
C, 60.41; H, 5.69; N,10.07
Found: C, 60.42; H, 5.62; N,10.10
EXAMPLE 4 w -
(4S)-cis-4-(3,4-Difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4-(2-
Cyano-phenyl)-cyclohexyl]-(3R)--pyrrolidin-3-yl}amide hydrochloride
_g4_
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F
CN
/ ~ N O / ~ F
N
H N
O~O ~ HC1
Analysis: Calcd. for C27H28F2N4O3 ~ HCl ~ 0.1 H20 ~ 0.95 EtOAc
C, 60.00; H, 6.02; N,9.09
Found: C, 60.02; H, 5.75; N,9.06
The following Examples were prepared by procedures
substantialy as described above for Examples 1-4 except substituting the
appropriate halozinc reagent (purchased from RIEKE Chemical) in step
B.
EXAMPLE 5
( 4S )-tran s-4-( 3, 4-Difluoro-phenyl )-3- ( 1-[4-( 2-ethoxycarbonyl-phenyl )-
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo-
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester
hydrochloride
F
/ ~"... N O / \ F
N N
C02Et ~~ ~ C02Me
N
O M a ~ HCl
Analysis: Calcd. for Cg4H40F2N4~5 ~ HCl ~ 0.75 H20 ~ 0.05 EtOAc
C, 58.01; H, 5.96; N,7.91
Found: C, 57.89; H, 5.99; N,8.31
EXAMPLE 6
{4S)-cis-4-(3,4-Difluoro-phenyl)-3-( 1-[4-{2-ethoxycarbonyl-phenyl )
cyclohexyl]-(3R)--pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo
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1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester
hydrochloride
F
O ~ ~ F
N
N N
C02Et p~ C02Me
N /
OMe ~ HCl
Analysis: Calcd. for C34H4oF2N40s ~ HCl ~ 0.75 H20 ~ 0.05 EtOAc
C, 57.20; H, 6.16; N,7.80
Found: C, 57.21; H, 6.00; N,8.46
EXAMPLE 7
(4S)-trans-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid-(1-
[4-(2-ethoxycarbonylphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide
hydrochloride
F
O ~ ~ F
", N
N
C02Et H N
O~O ~ HCl
Analysis: Calcd. for C2gH33F2N3~5 ~ HCl ~ 0.15 H20 ~ 0.05 EtOAc
C, 59.93; H, 5.98; N,7.18
Found: C, 59.94; H, 5.78; N,7.52
EXAMPLE 8
(4S)-cis-4-(3,4-Difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-(1-..[4-(2
ethoxycarbonylphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide
hydrochloride
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F
O / ~ F
N
N
C02Et
O O ~ HCl
Analysis: Calcd. for C2gH33F2N3~5 ~ HCl ~ 0.85 H20 ~ 0.05 EtOAc
C, 58.67; H, 6.09; N,7.03
Found: C, 58.63; H, 5.86; N,7.05
EXAMPLE 9
(4S)-trans-4-( 3,4-Difluorophenyl)-3-[ 1-(4-pyridin-2-yl-cyclohexyl )-(3R)
pyrrolidin-3-ylcarbamoyl]-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydro
pyrimidine-5-carboxylic acid methyl ester
F
N O / \ F
-N H N
/ C02Me
N
H OMe ~ HCt
Analysis: Calcd. for CgOH35F2N5~5 ~ HCl ~ 0.45 H20 ~ 0.2 EtOAc
C, 57.28; H, 6.01; N,10.84
Found: C, 57.25; H, 5.73; N,10.85
EXAMPLE 10
(4S)-cis-4-(3,4-Difluorophenyl)-3-[1-(4-pyridin-2-yl-cyclohexyl)-(3R)
pyrrolidin-3-ylcarbamoyl]-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydro
pyrimidine-5-carboxylic acid methyl ester
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F
F
N
-N H N
C02M a
N
H O M a ~ HCl
Analysis: Calcd. for CgpH35F2N5~5 ~ HCl ~ 0.35 EtOAc
C, 57.93; H, 6.01; N,10.76
Found: C, 58.17; H, 5.80; N,10.73
EXAMPLE 11
(4S)-trans-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid-[1
(4-pyridin-2-yl-cyclohexyl)-(3R)-pyrrolidin-3-yl]amide hydrochloride
F
~ "", O ~ ~ F
-N, ~N
H N
O~O ~ HC1
Analysis: Calcd. for C25H28F2N4O3 ~ HCl ~ 2.5 H20 ~ 0.05 EtOAc
C, 51.05; H, 6.02; N,9.45
Found: C, 50.90; H, 5.30; N,9.40
EXAMPLE 12
(4S)-cis-4-(3,4-Difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-[1-(4-
pyridin-2-yl-cyclohexyl)-(3R)-pyrrolidin-3-yl]amide hydrochloride
F
N O / ~ __
_ N N'\
H N
O~O ~ HCl
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Analysis: Calcd. for C25H28FZN4O3 ~ HCl ~ 1.65 H20 ~ 0.1 EtOAc
C, 55.92; H, 6.12; N,10.27
Found: C, 55.92; H, 5.85; N,10.22
Examples 13 - 17 were prepared according to Scheme 10.
EXAMPLE 13
(4S)-trans-4-{3,4-Difluorophenyl)-6-methoxymethyl-3-{ 1-[4-(2-methoxyl
phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-2-oxo-1,2,3,4
tetrahydro-pyrimidine-5-carboxylic acid methyl ester hydrochloride
F
OMe
~ F
~ ,.... N O
N N
/ C02Me
N
H OMe ~ HCl
Step A: 8-(2-Methoxy-phenyl)-1,4-dioxa-spiro[4.5]decan-8-of
To a solution of 2-Bromoanisole (1.87g, 10.0 mmol) in 10 mL
THF under Argon was added n-butyllithium (4.0 mL 2.5M in hexane,
I0.0 mmol) cooled to -?8°C. The solution was stirred 10 min. at -
78° C,
and a solution of 1,4-Cyclohexanedione mono-ethylene ketal ( 1.56g, 10.0
mmol) in 10 mL THF was added slowly. The solution was warmed to r.t.
and stirred for 60 min., then poured onto saturated sodium bicarbonate
and extracted with ethylacetate. The combined organic layers were
washed with saturated sodium chloride, dried with magnesium sulfate,
and concentrated in vacuo. The crude solid was crystallized with
hexane/diethylether 2:1 to give 8-(2-Methoxy-phenyl)-1,4-dioxa-
spiro[4.5]decan-8-of as a white solid.
1H NMR 8H (CDC13) 7.4-7.2 (m, 2H), 7.0-6.9 (m, 2H), 4.1-3.9
(m, 4H), 3.9 (s, 3H), 2.3-2.0 (m, 6H), 1.7-1.6 (m, 2H).
Step B: 4- (2-Methoxy-phenyl)-cyclohex-3-enone and 4-(2-Methoxy-
phenyl)-cyclohex-2-enone
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To a solution of 8-(2-Methoxy-phenyl)-1,4-dioxa-
spiro[4.5]decan-8-of (100mg, 0.379 mmol) in 10 mL THF was added HCl
(1.4 mL 6M, 8.4 mmol). The solution was heated to 80°C for 45 min, then
poured onto saturated sodium bicarbonate and extracted with
ethylacetate. The combined organic layers were washed with saturated
sodium chloride, dried with magnesium sulfate, and concentrated in
vacuo. The crude material was passed through silica (dichloromethane/
3% methanol as eluent) to give 4-(2-Methoxy-phenyl)-cyclohex-3-enone
1H NMR 8H (CDC13) 7.3-7.1 (m, 2H), 7.0-6.9 (m, 2H), 5.9-5.8
(m, 1H), 3.9-3.8 (s, 3H), 3.1-3.0 (m, 2H), 2.9-2.8 (m, 2H), 2.7-2.5 (m, 2H)
and 4-(2-Methoxy-phenyl)-cyclohex-2-enone.
1H NMR 8H (CDC13) 7.3-7.2 (m, 1H), 7.2-7.1 (m, IH), 7.0-6.9
(m, 3H), 6.2-6.1 (m, 1H), 4.2-4.1 (m, 1H), 3.8 (s, 3H), 2.6-2.4 (m, 2H), 2.4-
2.2 (m, 1H), 2.1-1.9 (m, 1H).
Step C: 4- (2-Methoxy-phenyl)-cyclohexanone
To a suspension of palladium on carbon (3.75 g, 25% wt) in
200 mL ethylacetate was added a 1:2 mixture of 4-(2-Methoxy-phenyl)-
cyclohex-3-enone and 4-(2-Methoxy-phenyl)-cyclohex-2-enone (15 g, 75.0
mmol) under argon. The suspension was stirred under a hydrogen at 1
atm. for 6.5 h, filtered through celite,concentrated in vacuo, and
crystallized from ethylacetate to give 4- (2-Methoxy-phenyl)-
cyclohexanone.
iH NMR 8g (CDC13) 7.3-7.1 (m, 2H), 7.0-6.9 (m, 2H), 3.88 (s,
1H), 3.6-3.4(m, 1H), 2.6-2.4 (m, 4H), 2.3-2.2 (m, 2H), 2.0-1.8 (m, 2H).
The title compound of Example 13 was prepared from the
product of step C by procedures identical to that described above for
EXAMPLE 1 Steps E + F.
trans-4-(3,4-Difluoro-phenyl)-6-methoxymethyl-3-( 1-[4-(2-methoxyl-
phenyl)-cyclohexyl]-3R-pyrrolidin-3-ylcarbamoyl)-2-oxo-1,2,3,4-
tetrahydro-pyrimidine-5-carboxylic acid methyl ester hydrochloride
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Analysis Calcd. for C32H3gF2N4Os ~ HCl ~ 0.45 H20
C, 58.56; H, 5.98; N, 8.54
Found: C, 58.55; H, 6.59; N, 8.15
The following Examples were prepared from the product of
step C above by procedures substantially as described above for Example
1 steps E + F.
EXAMPLE 14
(4S)-cis-4-(3,4-Difluoro-phenyl)-6-methoxymethyl-3-{1-[4-(2-methoxyl-
phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-2-oxo-1,2,3,4-
tetrahydro-pyrimidine-5-carboxylic acid methyl ester hydrochloride
F
F
O
C02Me
N N~N
OMe H O~.N OMe
H
~ HCl
Analysis: Calcd. for C32H3gF2N4O6 ~ HCl ~ 0.7 H20
C, 57.96; H, 6.25; N, 8.19
Found: C, 58.66; H, 6.05; N, 8.19
EXAMPLE 15
(4S)-trans-4-(3,4-Difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4-
(2-methoxyphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide hydrochloride
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F
F
O
N N' _N
OMe H ~O
//O
~ HCl
Analysis: Calcd. for C32H38F2N4O6 ~ HCl ~ 0.7 H20 ~ 0.25 EtOAc
C, 60.42; H, 5.69; N,10.07
Found: C, 60.42; H, 5.62; N,10.10
EXAMPLE 16
(4S)-cis-4-(3,4-Difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4-(2
methoxyphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide hydrochloride
F
F
O
N N~N
OMe H ~O
O
~ HCl
Analysis: Calcd. for C32H38F2N4O6 ~ HCl ~ 0.1 H20 ~ 0.95 EtOAc
C, 60.00; H, 6.02; N,9.09
Found: C, 60.02; H, 5.75; N,9.06
Examples 17 - 20 prepared according to Scheme 11. w -
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EXAMPLE 17
(4S)-cis-4-(3,4-Difluoro-phenyl) -3-[1-(4-hydroxy-4-pyridin-2-yl-cyclohexyl)
(3R)-pyrrolidin-3-ylcarbamoyl~-6-methoxymethyl -2-oxo-1,2,3,4
tetrahydro-pyrimidine-5-carboxylic acid methyl ester hydrochloride
F
HO O / ~ F
/~ N ~ -
,,
-N H N
/ C02Me
N
H O M a ~ HCl
Step A: 8-(2-Pyridyl)-1,4-dioxaspiro[4,5]decan-8-of
To a solution of 2-bromo-pyridine (10 g, 63. 3mmo1) in
tetrahydrofuran (200 mL) at -78 °C under an argon atmosphere was
added a solution of n-butyllithium ( 26 mL of 2.5N solution, 65 mmol).
The solution was stirred at -78°C for 10 minutes and then a
solution of
cyclohexanedione monoethylene ketal ( 10 g, 64 mmol) in
tetrahydrofuran(50 mL) was added. The reaction was allowed to warm to
room temperature and poured into a saturated solution of sodium
15 bicarbonate (mL) the mixture was extracted with ethyl acetate. The
combined ethyl acetate fractions were dried over anhydrous sodium
sulfate, filtered, and concentrated at reduced pressure. The residue was
chromatographed over silica gel eluting with 25% to 50% ethyl
acetate/hexane to give the product.
20 1H NMR. (CDC13~ 300 MHz) 8 8.5 (m, 1H), 7.69 (dt, J = 2, 7.5 Hz, 1H), 7.40
(dt, J = 7.5, 1 Hz, 1H), 7.2 (ddd, J = 1.5, 4.5, 7 Hz, 1H), 5.25 (s, 1H ex),
4.0
(m, 4H), 2.3-2.0 (m, 4H), 1.8-1.65 (m, 4H)..
Step B: 4-Hydroxy-4-(2-pyridyl)-cyclohexanone
_ __
A solution of the product of step A (8-(2-pyridyl)-1,4-
dioxaspiro[4,5]decan-8-ol) ( 8.3 g, 35.5 mmol) in dioxane (150 mL) was
treated with 6 N HCl ( 60 mL) and stirred at room temperature for 0.5 hr.
The reaction was carefully poured into cold saturated sodium
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bicarbonate (1 L). The mixture was extracted with ethyl acetate (3 X 200
mL) and the combined extracts were dried over anhydrous sodium
sulfate, filtered, and concentrated at reduced pressure to give 5.3 g of the
product.
1H NMR (CDCl3~ 300 MHz) 8 8.6 (br d, 1H), 7.8-7.7 (m, 1H),
7.35 (d, J = 5 Hz, 1H), 7.3-7.25 (m, 1H), 5.52 (s, 1H ex), 3.2-2.95 (m, 2H),
2.48-2.35 (m, 2H). 2.35-2.20 (m, 2H). 2.1-2.0 {m, 2H).
The title compound of Example 17 was prepared from the
product of step B by procedures identical to those described above for
EXAMPLE 1 Steps E + F
Analysis Calcd. for C3pH35F2N5~6 ~ HCl ~ 0.1 H20
C, 58.94; H, 5.99; N,11.46
Found: C, 58.59; H, 5.59; N,11.24
The following Examples were prepared from the product of
step B above by procedures substantially as described above for Example
1 steps E + F.
EXAMPLE 18
(4S)-trans-4-(3,4-Difluoro-phenyl) -3-[1-(4-hydroxy-4-pyridin-2-yl
cyclohexyl)-(3R)--pyrrolidin-3-ylcarbamoyl]-6-methoxymethyl -2-oxo
1,2,3,4-tetrahydro-pyrimidine-5-carboxylic acid methyl ester
hydrochloride
F
HO ~ ~ F
N~ ~
-N H N
O~ ~ C02Me
N ._
H OMe ~ HCl
Analysis: Calcd. for CgpHg5F2N5~6 ~ HCl ~ 0.55 H20 ~ O.1CH2C12
C, 58.87; H, 5.83; N,11.39
Found: C, 58.88; H, 5.68; N,11.25
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EXAMPLE 19
(4S)-cis-4-{3,4-Difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-[1-{4
hydroxy-4-pyridin-2-yl-cyclohexyl)-(3R)--pyrrolidin-3-yl]amide
hydrochloride
F
HO ~ \ F
\ ,,,, N O
N' \
-N H N
O~O ~ 2 HCl
Analysis: Calcd. for C25H28F2N5O4 ~ 2 HCl ~ 0.05 H20 ~ 0.25 EtOAc
C, 53.62; H, 5.56; N,9.62
Found: C, 54.30; H, 5.66; N,9.60
EXAMPLE 20
(4S)-trans-4-(3,4-Difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-[1-(4
hydroxy-4-pyridin-2-yl-cyclohexyl)-(3R)-pyrroiidin-3-yl]amide
hydrochloride
F
HO ~ \ F
\ N I O _
\~N' \
N H N
O- O ~ 2 HCl
Analysis: Calcd. for C25H28F2N5O4 ~ 2 HCl ~ 0.3 H20 ~ 0.25 EtOAc
C, 53.21; H, 5.60; N,9.55
Found: C, 54.18; H, 5.57; N,9.29
Examples 21 - 24 were prepared according to Scheme 13.
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EXAMPLE 21
(4S)-4-(3,4-Difluoro-phenyl)-3-( 1-[4-(2-hydroxy-phenyl)-cyclohexyl]-(3R)-
pyrrolidin-3-ylcarbamoyl)-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester
F
N O / \ F
N~N
OH ~~ ~ C02Me
N
H OMe
Step A: 8-(2-Benxyloxy-phenyl)-1,4-dioxa-spiro[4.5]decan-8-of
To a solution of 8-(2-Hydroxy-phenyl)-1,4-dioxa-
spiro[4.5]decan-8-of (2.Og, 7.9 mmol) in 10 mL DMF was added
potassium carbonate (3.91g, 28.3 mmol) and benzyl bromide (1.35g, 1.9
mmol) under argon. The suspension was stirred for 1.5 h, poured onto
10% potassium hydrogen sulfate, and extracted with ethylacetate. The
combined organic layers were washed with saturated sodium chloride,
dried with magnesium sulfate, and concentrated in vacuo to give the
product (2.76g, 100%).
1H NMR &H (CDC13) 8.02 (bs,lH), 7.45-7.35 (m, 2H), 7.25-7.20
(m, 2H), 7.05-6.95 (m, 1H), 5.18 (s, 2H), 4.00-3.90 (m, 4H), 2.20-2.10 (m,
4H), 1.70-1.60 (m, 4H).
Step B: 4-(2-Benzyloxy-phenyl)-4-hydroxy-cyclohexanone
To a solution of of 8-(2-Hydroxy-phenyl)-1,4-dioxa-
spiro[4.5]decan-8-of (2.76 g, 8.12 mmol) in 70 mL 1,4-dioxane was added
aqueous hydrogen chloride (40.6 mL 1M, 40.6 mmol). The solution was
stirred for 30 min., poured onto saturated sodium bicarbonate, and
extracted with ethylacetate. The combined organic layers were washed
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with saturated sodium chloride, dried with magnesium sulfate, and
concentrated in vacuo to give the product (1.96 g, 82%).
1H NMRBH (CDC13) 7.35-7.25 (M, 2H), 7.10-7.00 (m, 2H), 5.20
(s, 2H), 4.44 (s, 1H), 3.00-2.85 (m, 2H), 2.55-2.40 (m, 2H), 2.35-2.15 (m,
4H).
Step C: 4-(2-Benzyloxy-phenyl)-cyclohex-3-enone and 4-(2-Benzyloxy-
phenyl)-cyclohex-2-enone
To a solution of 4-(2-Benzyloxy-phenyl)-4-hydroxy-
cyclohexanone (1.9 g, 6.42 mmol) in 70 mL benzene was addedp-
toluenesulfonic acid monohydrate (0.070 g, 0.368 mmol). The solution
was warmed to 60°C, stirred for 1 h, then poured onto saturated sodium
bicarbonate, and extracted with ethylacetate. The combined organic
layers were washed with saturated sodium chloride, dried with
magnesium sulfate, and concentrated in vacuo to give a mixture of
enone products (1.7 g, 100%).
1H NMR &H (CDC13) 7.45-7.20 (m, 7H), 7.00-6.90 (m, 2H), 5.83
(s, 1H), 5.13 (s, 2H), 3.03 (bs, 2H), 2.83 (bt, 2H, J 6.35 Hz), 2.52 (bt, 2H,
J
6.83).
Step D: 4-(2-Hydroxy-phenyl)-cyclohexanone
To a solution of a mixture of 4-(2-Benzyloxy-phenyl)-
cyclohex-3-enone and 4-(2-Benzyloxy-phenyl)-cyclohex-2-enone (1.7 g,
6.12 mmol) in 25 mL ethylacetate under argon was added 10% palladium
on carbon (0.425 g, 25 wt. %). The suspension was stirred under
hydrogen at 1 atm. for 24 h, filtered through celite, and concentrated in
vacuo to give the product (1.13 g, 97%).
1H NMR &~ (CDC13) 7.20-7.05 (m, 2H), 6.92 (t, 1H, J 7.56 Hz),
6.75 (d, 1H, J 8.06 Hz), 3.35-3.45 (m, 1H), 2.65-2.45 (m, 4H), 2.30-2.20 (m,
2H), 2.0- 1,85 (m, 2H).
The title compound of Example 21 was prepared from the
product of step D by procedures identical to that described in EXAMPLE
1 Steps E + F
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Analysis Calcd. for CglHggF2N506 ' 0~9 H20 ~ 0.4 CH2C12
C, 58.12; H, 6.00; N, 8.64
Found: C, 58.21; H, 5.72; N, 8.66
The following examples were prepared from the product of
step D above by procedures substantially as described above for Example
1 steps E + F.
EXAMPLE 22
(4S)-cis-4-(3,4-Difluoro-phenyl)-6-methoxymethyl-3-{ 1-[4-(2-hydroxy-
phenyl)-cyclohexyl]-(3R)--pyrrolidin-3-ylcarbamoyl)-2-oxo-1,2,3,4-
tetrahydro-pyrimidine-5-carboxylic acid methyl ester
F
/ ~ F
/ v N~ ~
N N
OH ~~ / C02Me
N
H OMe
Analysis: Calcd. for CglHg6F2N5~6 ' 0.4 H20 ~ 0.1 CH2C12
C, 61.00; H, 5.76; N, 9.15
Found: C, 61.01; H, 5.77; N, 8.91
EXAMPLE 23
(4S)-trans-4-(3,4-Difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4-
(2-hydroxyphenyl)-cyclohexyl]-(3R)--pyrrolidin-3-yl)amide
F
/ ~ ,.... N F . __
N~
OH H
O
Analysis: Calcd. for C2gH29F2N3~4 ' 0.5 H20 ~ 0.2 CH2C12
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C, 61.50; H, 5.99; N, 8.22
Found: C, 61.42; H, 5.76; N, 8.32
EXAMPLE 24
(4S)-cis-4-(3,4-Difluorophenyl)-2-oxooxazolidine-3-carboxylic acid-{1-[4-(2-
hydroxyphenyl)-cyclohexyl]-(3R)--pyrrolidin-3-yl}amide
F
N O / ~ F
N' \
OH H N
O~ _O
Analysis: Calcd. for C26H29F2N3~4 ~ 0.3 H20 ~ 0.15 CH2CI2
C, 62.35; H, 5.98; N, 8.34
Found: C, 62.73; H, 5.81; N, 8.30
Utilizing the methodology described in detail herein, the
following additional compounds shown in Tables 1-3 were prepared.
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N 1-'
rr
(Jt
H
A
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v
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CA 02293408 1999-12-06
WO 98/57641 PCT/US98/12673
w '~~"+ ~,
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CA 02293408 1999-12-06
WO 98/57641 PCT/US98/12673
O
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-102 -
CA 02293408 1999-12-06
WO 98/57641 PCT/US98/12673
x a~
0 0
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n
-103 -
CA 02293408 1999-12-06
WO 98/57641 PCT/US98/12673
d d ~ x a
b
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-104 -
CA 02293408 1999-12-06
WO 98/57641 PCT/US98/12673
0
'Ti TI
n
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-105 -
CA 02293408 1999-12-06
WO 98/57641 PCT/US98/12673
O
n
n n
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a
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EXAMPLE 25
(4S)-trans-4-(3-,4-Difluorophenyl)-3-{ 1-[4-(2-fluorophenyl)-cyclohexyl]-
(3R)-pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester hydrochloride
7 H ~HCI
F
F
Step A: 8-(2-Fluorophenyl)-1,4-dioxa-spiro[4.5]decan-8-of
To a solution of 2-fluoro-1-Iodobenzene (ll.lg, 50.0 mmol) in
50 mL THF, cooled to -78°C under argon, was added n-butyllithium (20.0
mL 2.5M in hexane, 50.0 mmol). The solution was stirred 10 min. at -78°
C, and a solution of 1,4-cyclohexanedione mono-ethylene ketal (7.81g,
50.0 mmol) in 50 mL THF was added slowly. The solution was warmed
to r.t. and stirred for 60 min., then poured onto saturated sodium
bicarbonate and extracted with ethylacetate. The combined organic
layers were washed with saturated sodium chloride, dried with
magnesium sulfate, and concentrated in vacuo. The crude solid was
crystallized from dichloromethane / hexane to give 8-(2-Fluoro-phenyl)-
1,4-dioxa-spiro-[4.5]decan-8-of (3.8g, 30°l0).
'H NMR 8 (CDC13) 7.54-7.47 (m, 1H), 7.28-7.20 (m, 1H), 7.15-
7.00 (m, 2H) 4.01 (s, 4H), 2.39-2.28 (m, 2H), 2.18-2.07 (m, 2H), 1.95 -1.85-
(m, 2H), 1.75-1.60 (m, 2H).
Step B: 4-(2-Fluorophenyl)-4-hydroxy-cyclohexanone
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To a solution of 8-(2-fluorophenyl)-1,4-dioxa-spiro[4.5]decan-
8-0l (5.8 g, 23 mmol) in dioxane (100 mL), cooled to 0°C was added
hydrogen chloride (1M in water, 75 ml, 75 mmol). The reaction mixture
was stirred for 2 hour at room temperature, extracted with ethyl ether
(2x 200 mL) and washed with saturated bicarbonate solution, water and
brine. Drying and solvent evaporation gave 4-(2-Fluorophenyl)-4-
hydroxy-cyclohexanone in quantitative yield.
1H NMR (CDC13) 1H NMR d (CDCl3) 7.60-7.50 (m, 1H), 7.40-7.00 (m, 3H),
3.0-2.8 (m, 2H), 2.6-2.3 (m, 4H), 2.3-2.2 (m, 2H).
Step C : 4- (2-Fluorophenyl)-cyclohexanone:
4-(2-Fluorophenyl)-4-hydroxy-cyclohexanone from Step B
was added to trifluoroacetic acid at 0°C and the reaction was stirred
for
30 minutes and then poured onto saturated sodium bicarbonate and
extracted with ethylacetate. The combined organic layers were washed
with saturated sodium chloride, dried with magnesium sulfate, and
concentrated in vacuo. to give the mixture of olefins, 4- (2-Fluorophenyl)-
cyclohex-3-enone and 4-(2-Fluorophenyl)-cyclohex-2-enone (5.2 g)
To a suspension of palladium on carbon (1.1 g) in 100 mL
ethylacetate was added the mixture of 4-(2-Fluoro-phenyl)-cyclohex-3-
enone and 4-(2-Fluorophenyl)-cyclohex-2-enone (5.1 g) under argon. The
suspension was placed under 50 psi hydrogen for 36 h, filtered through
celite, concentrated in vacuo, and passed through silica (10%
ethylacetate,hexane) to give 4-(2-fluorophenyl)-cyclohexanone (3.5 g).
tH NMR ~, (CDC13) 7.29-7.20 (m, 2H), 7.15-7.04 (m, 2H), 3.44-
3.34 (m, 1H), 2.64-2.41 (m, 4H), 2.26-2.21 (m, 2H), 2.06-2.01 (m, 2H).
Step D: cis and trans -1-[4-(2-Fluorophenyl)-cyclohexyl]-pyrrolidin-
3-ylamine dihydrochloride
To a solution of 4- (2-Fluorophenyl)-cyclohexanone (1.2 g,
6.25 mmol) and acetic acid (1.3 mL 22.75 mmol) in 30 mL methanol was
addded (3R)-(+)-3-(tent-butoxycarbonyiamino)-pyrrolidine (1.74 g, 9.38
mmol) under argon. The solution was stirred for 2h, then sodium
cyanoborohydride (9.36 mL 1M in THF, 9.38 mmol) was added dropwise.
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The solution was poured onto saturated sodium bicarbonate, and
extracted with ethylacetate. The combined organic layers were washed
with saturated sodium chloride, dried with magnesium sulfate, and
concentrated in vacuo. The crude material was passed through silica
5 (3% methanol, ethylactate) to give 2:1 trans:cis 1-tertbutoxycarbonyl-[4-
(2-fluoro-phenyl)-cyclohexyl]pyrrolidin-3-ylamine (2.3 g, 100°~0). The
trans-1-tertbutoxycarbonyl-[4-(2-fluoro-phenyl)-cyclohexyl]pyrrolidin-3-
ylamine (1.53 g, 4.23 mmol) was dissolved in 100 mL ethylacetate, and
HCl gas was bubbled through the solution for 20 min. The solution was
10 concentrated in vacuo to give tracns -1-[4-(2-Fluorophenyl)-cyclohexyl]-
pyrrolidin-3-ylamine dihydrochloride (1.4 g, 100%).
'H NMR 8H (CD30D) 7.31-7.26 (m, 2H), 7.29-7.03 (m, 2H), 4.15
(m, 1H), 3.90-3.45 {m, 2H), 3.42-3.25 (m, 2H), 3.0-2.85 (m, 1H), 2.70-2.45
(m, 1H), 2.40-2.20 {m, 2H), 2.05-2.02 (d, 2H, J= 9.5 Hz), 1.79-1.67 (m, 4H).
Step E: trans-4-{3-,4-Difluorophenyl)-3-(1-[4-(2-fluorophenyl)-
cyclohexyl]-pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-
1,2,3,4-tetrahydro-pyrimidine-5-carboxylic acid methyl ester
hydrochloride
To a solution of trans-1-[4-(2-fluoro-phenyl)-cyclohexyl]-
pyrrolidin-3-ylamine dihydrochloride (100 mg, 0.299 mmol) and
triethylamine (0.125 mL, 0.897 mmol) in 3.0 mL DMF was added (+)-3-(4-
nitrophenoxycarbonyl)-4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-
25 1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester (477 mg,
0.329 mmol) under argon. The solution was stirred for 2h, poured onto
saturated sodium bicarbonate, and extracted with ethylacetate. The
combined organic layers were washed with saturated sodium chloride,
dried with magnesium sulfate, and concentrated in vacuo. The crude
30 material was passed through silica (3% methanol, dichloromethane)
and washed with sodium carbonate. To a solution of the free base in
ethylacetate was added excess hydrogen chloride (1M in diethylether).
This solution was concentrated in vacuo to give the product ( 133 mg,
67%).
Analysis: Calcd. for C3,H35F3N4O5~HCl~O.55 HZO
C, 57.54; H, 5.78; N, 8.66.
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Found: C, 57.58; H, 5.91; N, 8.79
The compounds set forth in Examples 26-42 below were
prepared by procedures substantially the same as described above in
Example 25. Additional preparative details are provided in certain
Examples as deemed appropriate.
EXAMPLE 26
(4S)-trczns-4-(3,4-Difluoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid -{ 1-
(4-(2-fluoro-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}-amide
hydrochloride
O
-O
C~ N
?l-NJ
O
~HCI
F
F
Analysis: Calcd. for C3,H35F3N405~HCl~0.35 Hz0~0.20 EtOAc
C, 58.74; H, 5.76; N, 7.67
Found: C, 58.74; H, 5.82; N, 7.50
EXAMPLE 27
(4S)-tracns-4-(3,4-Difluoro-phenyl)-3-{1-[4-(4-fluoro-2-methoxy-phenyl)-
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo-
1,2,3,4-tetrahydro-pyrimidine-5-carboxylic acid methyl ester
hydrochloride
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F,
O
O-
F
Analysis: Calcd. for C32Ha7FaN406 ~HCl ~ 1.35 H20 ~ 0.20 EtOAc
C,57.00; H, 6.32; N, 8.11
Found: C, 57.03; H, 5.95; N, 7.88
EXAMPLE 28
(4S)-trans-4-(3,4-Difluoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid (1
10 [4-(4-fluoro-2-methoxy-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl)-amide
hydrochloride
O
N
F ?r-~J
O
~i
F
F
15 Analysi s : Calcd. for C27H3oF3N304 ~ HC1 ~ 0.40 Hz0 ~ 0.45 EtOAc
C,57.56; H, 5.94; N, 6.99
Found: C, 57.56; H, 5.68; N, 7.00
EXAMPLE 29
20 (4S)- 4-(3,4-Difluoro-phenyl)-3-(1-[4-(2-fluoro-phenyl)-4-hydroxy-
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl)-6-methoxymethyl-1,2,3,4-
tetrahydro-pyrirnidine-5-carboxylic acid methyl ester hydrochloride
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H3C0 H ~HCI
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Prepared from the product of Example 25, Step B by procedures similar
to those described for Steps D and E of Example 25.
F
7 ~H ~HCI
F
Analysis: Calcd. for C31H35F3N406~HCl~1.00 HZO~0.05 ether
C,55.53; H,5.75; N, 8.30
Found: C, 55.53; H, 5.60; N, 8.34
EXAMPLE 30
(4S)-4-(3,4-Difluoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid -{1-[4-(2-
fluoro-phenyl)-4-hydroxy-cyclohexyl)-(3R)-pyrrolidin-3-yl}-amide
hydrochloride
O,'
N ~O
N
~J
O
F OH ~HCI ~ I
F
F
Analysis: Calcd. for CZSH2gF3N304~HCl~0.40 HZO
C,57.07; H, 5.49; N,7.68
Found: C,57.03; H, 5.32; N, 7.42
EXAMPLE 31
(4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid {1-[4-
hydroxy-4-(2-trifluoromethylphenyl)-cyclohexyl)-(3R)-pyrrolidin-3-yl}
amide
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F
C F3 l ~ F
~ O
N II
HO N~N
H
O O
Analysis: Calculated for C17H28N304F5 ~ 0.15 EtOAc:
C, 58.49; H, 5.19; N, 7.41
Found: C, 58.50; H, 4.98; N, 7.13
EXAMPLE 32
(4S)-4-(3,4-Difluorophenyl)-3-{1-[4-hydroxy-4-(2-trifluoromethylphenyl)
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester
F
F
C F3
N OI[
HO N~N COOMe
H ~ I OMe
O N
H
Analysis: Calculated for C32Hs5N40sFs ' 0.30 EtOAc:
C, 57.53; H, 5.44; N, 8.08
Found: C, 57.92; H, 5.43; N, 7.82 - -
EXAMPLE 33
(4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid {1-[4-(2
trifluoromethylphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl} amide
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F
C F3 ' ~ F
O
N
N ~N
H
O O
Analysis: Calculated for CZ7H28N3O3F5:
C, 60.32; H, 5.26; N, 7.82
Found C, 60.36; H, 5.29; N, 7.50
EXAMPLE 34
(4S)-4-(3,4-Difluorophenyl)-6-methoxymethyl-2-oxo-3-{1-[4-(2-
trifluoromethylphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester
F
F
C F3
\ O
N II
N~N COOMe
OMe
O N
Analysis: Calculated for C~ZH3~N405F5 ~ 0.15 CHC13:
C, 57.75; H, 5.30; N, 8.38
Found: C, 57.53; H, 5.30; N, 8.51 - -
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EXAMPLE 35
(4S)-4-(3,4-Difluorophenyl)-3-(1-[4-(4-fluorophenyl)-4-hydroxy-cyclohexyl]
(3R)-pyrrolidin-3-ylcarbamoyl)-6-methoxymethyl-2-oxo-1,2,3,4
tetrahydropyrimidine-5-carboxylic acid methyl ester hydrochloride
F
F
F / \ N O
COOMe
HO H N
O' _N OMe
HCI H
Step A.: 8-(4-Fluorophenyl)-1,4-dioxa-spiro[4.5]decan-8-of
To a solution of 1,4-cyclohexanedione mono-ethylene ketal (25 g, O.I6
mol) in diethylether (750 ml), cooled to -78°C was added 4-fluorophenyl
magnesium bromide {1 M in tetrahydrofuran, 190 ml, 0.19 mol)
dropwise. The reaction mixture was warmed to room temperature,
quenched with water and extracted twice with ethyl acetate. Washing
with brine, drying and solvent evaporation gave 8-(4-fluorophenyl)-1,4-
dioxa-spiro[4.5]decan-8-of {38 g, 95%).
'H NMR (CDC13) 8 7.55-7.40 (m, 2H) 7.05-6.95 (m, 2H), 3.95 (m, 4H), 2.20-
1.90 (m, 4H), 1.90-1.60 (m, 4H), 1.50 (s, 1H)
Step B: 4-(4-Fluorophenyl)-4-hydroxy-cyclohexanone
To a solution of 8-(4-fluorophenyl)-1,4-dioxa-spiro[4.5]decan-8-of
(38 g, 0.15 mol) in dioxane (900 ml), cooled to 0°C was added hydrogen
chloride (1M in water, 1.5 L, 1.5 mol). The reaction mixture was stirred
for 2 hour at RT and extracted twice with ethyl acetate. The ethyl acetate
layer was washed with saturated sodium bicarbonate solution, water-
and brine. Drying and solvent evaporation gave 4-(4-fluorophenyl)-4-
hydroxy-cyclohexanone {24.8 g, ?9 %); 'H NMR (CDCl3) 8 7.55-7.45 (m,
2H) 7.16-7.0 (m, 2H), 3.0-2.85 (m, 2H), 2.18-2.38 (m, 6H), 1.85 (s, 1H)
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Step C: 4-(3-(3R)-tert-Butoxycarbonylamino-pyrrolidin-1-yl)-1-(4-
fluorophenyl)- cyclohexanol
HN
NHBoc
HO~~~ NaCNBH3, MeCOOH HO~
V\'O MeOH ~\N
NHBoc
To a solution of 4-(4-Fluorophenyl)-4-hydroxy-cyclohexanone
(2.0 g, 9.6 mmol) and acetic acid (2.6 ml, 48 mmol) in methanol (60 ml)
was added (3R)-(+)-3-(tert-butoxycarbonylamino) pyrrolidine (2.6 g, 14.4
mmol). After stirring for 2 hours at room temperature, sodium
cyanoborohydride (1M in tetrahydrofuran, 14.4 ml, 14.4 mmol) was
added dropwise. The reaction mixture was stirred for 45 minutes,
concentrated, diluted with 10% potassium hydrogen sulfate and
extracted twice with ether. The aqueous solution was basified with 1N
sodium hydroxide and extracted with ethyl acetate three times.
Washing with brine, drying and solvent evaporation gave an oil. Flash
chromatography (silica gel, 2-propanol-ethyl acetate, 4:96 increasing to
8:92) gave 4-(3-(3R)-tert-Butoxycarbonylamino-pyrrolidin-1-yl)-1-(4-
fluorophenyl)- cyclohexanol (2.0 g, 56%); 'H NMR (CDC13) 8 7.46(m, 2H),
7.02 (m, 2H), 4.89 (m, 1H), 4.18 (m, 1H), 2.94 (m, 1H), 2.71 (m, 2H), 2.42
(m, 1H), 2.26 (m, 1H), 2.13 (m, 1H), 1.83 (m, 9H), 1.63 (m, 1H), 1.44 (s,
9H),
A less polar isomer was also isolated.
Step D: 4-[(3R)-3-Aminopyrrolidin-1-yl]-1-(4-fluorophenyl)-
cyclohexanol dihydrochloride
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\
/ /.
HO~~ HC~ HO
~ \N EtOAc N
NHBoc 2 HCI NH2
To a solution of 4-((3R)-3-tert-Butoxycarbonylamino-pyrrolidin-1-
yl)-1-(4-fluorophenyl) cyclohexanol (1.05 g, 2.8 mmol) in ethyl acetate (50
ml), cooled to 0°C was added hydrogen chloride gas, bubbled in gently
for
5-10 minutes. The reaction mixture was stirred for 20 minutes at 0°C
and the above procedure was repeated twice. Concentration and
flushing with ethyl acetate three times gave 4-((3R)-3-aminopyrrolidin-1-
yl)-1-(4-fluorophenyl) cyclohexanol dihydrochloride (0.87 g, 89%); 'H
NMR (DMSO) 8 11.33 (bd, 1H) ), 8.60 (bd, 3H), 7.52 (m, 2H), 7.14 (m,2H),
5.12 (bs, 1H), 3.35-4.04 (m, 6H), 1.76-2.27 (m, lOH).
Step E: 4-(3,4-Difluorophenyl)-3-{1-[4-(4-fluorophenyl)-4-hydroxy
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6
methoxymethyl-1,2,3,4-tetrahydro-pyrimidine-5-carboxylic
acid methyl ester hydrochloride was prepared by a
procedure substantially as described above for the
preparation of Example 25
Mass Spectrum: 617.4 (M+1), FAB
EXAMPLE 36
(4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid{ 1-[4-(4-
fluorophenyl)-4-hydroxy-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide _ __
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F
\ F
F / \ N O
HO N~N
H
HCI O
Mass Spectrum: 504.3 (M+1), FAB
EXAMPLE 37
(4S)-4 -(3,4-Difluorophenyl)-3-~1-(4-(4-fluorophenyl)-4-hydroxy
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6-methyl-2-oxo-1,2,3,4
tetrahydropyrimidine-5-carboxylic acid methyl ester hydrochloride
F
F
F / \ N O
COOMe
HO H N
HCI O' _N CH3
H
Analysis: Calculated for C3oH33N4~5F3 ' HCL ~ 1.5 H20:
C, 55.43; H, 5.74; N, 8.62
Found: C, 55.43; H, 5.48; N, 8.5fi
EXAMPLE 38 w
(4S)-4 -(3,4-Difluorophenyl)-3-(1-(4-(4-fluorophenyl)-cyclohexyl]-(3R)
pyrrolidin-3-ylcarbamoyl}-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5
carboxylic acid methyl ester hydrochloride
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F
F
/
F ~ \ N O
COOMe
H ~N I
O' _N CH3
HCI H
Analysis: Calculated for C3oH33N4~4F3 ~ HCL ~ 0.75 H20:
C, 58.06; H, 5.77; N, 9.03
Found: C, 58.09; H, 5.90; N, 8.86
EXAMPLE 39
(4S)-4 -(3,4-Difluorophenyl)-3-(1-[4-(4-fluorophenyl)-cyclohexyl]-(3R)
pyrrolidin-3-ylcarbamoyl)-6-methoxymethyl-2-oxo-1,2,3,4
tetrahydropyrimidine-5-carboxylic acid methyl ester hydrochloride
F
F
F ~ \ N O
COOMe
H ~ I OMe
HCI O N
H
Analysis: Calculated for C3,Ha5N4O6F3 ~ HCL ~ 0.45 H20:
C, 57.70; H, 5.76; N, 8.68
Found: C, 57.77; H, 5.75; N, 8.29
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EXAMPLE 40
(4S)-4 -(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid{1-[4-(4
fluorophenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}amide
F
\ F
F ~ ~ N O
N~N
H
O O
Analysis: Calculated for CZSHzgN3O3F3:
C, 64.04; H, 5.80; N, 8.62
Found: C, 63.93; H, 5.79; N, 8.58
EXAMPLE 41
(4S)-4 -(3,4-Difluorophenyl)-3-{1-[4-(4-fluorophenyl)-4-hydroxy-
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-2-oxo-1,2,3,4
tetrahydropyrimidine-5-carboxylic acid methyl ester
F
F
F / \ N O
COOMe
HO H N
O' _ N H
H
Analysis: Calculated for CZ9H3,N4O5F3 ~ 0.30 CHCI3:
C, 57.84; H, 5.19; N, 9.21
Found: C, 57.95; H, 5.54; N, 8.92
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EXAMPLE 42
(4S)-3-( 1-[4-(4-Cyanophenyl)-4-hydroxycyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl}-4 -(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester hydrochloride
F
F
/ \ O
NC N ~ COOMe
HO H N
O' _N OMe
HCI H
Analysis: Calculated for C32H35N50~F2 ' HCL ~ 0.75 H20:
C, 57.05; H, 5.61; N, 10.40
Found: C, 56.75; H, 5.68; N, 10.79
EXAMPLE 43
5-Cyclopropyl-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid
( 1-[4-(2-fluorophenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl-amide
F
F
F O
/ \ N N N
H O __
O
to A
2-(3,4-difluorophenyl)-3-cyclopropyl-3-hydroxy-propionic acid
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A solution of 3,4-difluorophenylacetic acid (2.9 g, 16.8 mmol) in 80 mL
dry THF was cooled to -78 °C and treated with LDA (2.0 M
heptane/THF/ethylbenzene, 42 mmol, 21 mL) for 15 minutes. Then
cyclopropylcarboxaldehyde (17 mmol, 1.27 mL, 1.19 g) was added via
syringe and the reaction warmed slowly to room temperature over 1 hr. .
The reaction mixture was diluted with 5 % aqueous potassium
hydrogen sulfate (50 mL), and extracted with ethyl acetate (2 x 150 mL}.
The combined extracts were dried (Na2S04), filtered and concentrated in
vacuo. The lower rf material crystallized to give 1.4 g of solid.
1H NMR (CDCl3, 300 MHz) 8 7.40 - 7.00 (m, 3H, Ar-H), 3.75-3.60 (br m,
1H), 3.50-3.40 (m, 1H), 0.95-0.70 (m, 1H), 0.5-0.20 (m, 4H)
Step B
4-(3,4-difluorophenyl)-5-cyclopropyl-oxazolidin-2-one
The resulting hydroxy acids (7.95 g, 32 mmole) without further
purification, were dissolved in dry degassed DMF (100 mL) and treated
with solid NaHC03 (14.0 g) and diphenylphosphorylazide (DPPA, 9.9 g,
36 mmol, 7.74 mL) at room temperature for 1 hr. The reaction was then
heated on a steam bath for 15 min. Signs of nitrogen evolution were
immediately apparent. The reaction mixture was poured into saturated
sodium bicarbonate and extracted with ethyl acetate. The organics were
dried over anhydrous magnesium sulfate, filtered and concentrated in
vacuo and purified by chromatography (Si02, 8 mm, 0 - 50%
EtOAc/hexanes) affording the (~)-traps diastereomer (4.9 g) followed by
the (~)-cis diastereomer (1.5 g). The traps isomer was resolved by HPLC
on a Chiralcel OD eluting with 10% ethanol hexanes containing I%
diethylamine.
For the cis isomer.
1H NMR (CDC13, 300 MHz) 8 7.35 - 7.20 (m, 2H, Ar-H), 7.10-7.05 (br m,
1H, Ar-H), 5.70 (br s, 1H, NH), 4.95 (d, 1H, J = 7.5 Hz, CH(Ar)}, 4.10 (t,
1H, J = 7.5 Hz, CHcycpr), 0.6-0.50 (m, 1H), 0.5-0.4 (m, 1H), 0.4-0.3 (m,
1H), 0.2-0.1 (m, 1H) _ _
For the trc~ns isomer:
IH NMR (CDC13, 300 MHz) 8 7.30 - 7.15 (m, 2H, Ar-H), 7.15-7.08 (br m,
1H, Ar-H), 5.85 (br s, 1H, NH), 4.75 (d, 1H, J = 7.2 Hz, CH(Ar)), 3.65 (t,
1H, J = 7.2 Hz, CHcycpr), 1.3-1.15 (m, 1H), 0.75-0.6 (m, 1H), 0.5-0.4 (m,
1H), 0.22-0.14 (m, 1H)
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Sin C
4-(3,4-Difluorophenyl)-5-cyclopropyl-2-oxo-oxazolidine-3-carboxylic acid-
s 4-vitro-phenyl ester
To a solution of 4-(3,4-difluorophenyl)-5-cyclopropyl-oxazolidin-2-one (2
g, 8.3 mmol) in 100 mL THF was added a solution of n-butyllithium in
hexane (9.1 mmol) dropwise via a syringe under an argon atmosphere at
-78°C. The resulting yellow solution was stirred at -78°C for 10
min. To
this solution was then added dropwise via syringe 4-
nitrophenylchloroformate (1.03 g, 5.1 mmol) in 20 mL of THF. The
reaction was stirred at -78° for 10 min. The reaction mixture was
poured
into saturated sodium bicarbonate aad extracted with ethyl acetate (2 X
200 mL). The organic extracts were washed with brine, and the organic
layer was dried over NaZSO,. The solvent was removed after filtration,
and the residue was purified by column chromatography on silica gel
30°k ethyl acetate hexane. The material was rechromatographed on
silica gel eluting with 2% acetone/ methylene chloride to give 1.2 g the
product as a thick syrup which solidified upon standing.
'H NMR (CDCh, 300 MHz) S 8.22 (d, J = 9.0 Hz, 2 H), 7_34 (d, J s 9.0 Hz, 2
H), 7.35-7.05 (m, 3 H), b.19 (d, J = 6.0 Hz, 1 H),3.80 (dd, J = 5.0,8.fi Hz, 1
H), 1.35-1.15 (m, 1H), 0.85-0.7 (m, 1H), 0.6-0.5 (m, 1H), 0.35-0.2 (m, 1H)
StemD.
b-cyclopropyl-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid
( 1-[4-(2-fluorophenyl)-cyclohexyl]-pyrrolidin-3-yl-amide
The title compound was prepared from the reaction of the product of Step
C above and traps -1-[4-(2-fluoro-phenyl)-cyclohexyl~-pyrrolidin-3-
ylamine dihydrnchloride
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F
F
F O
N~ N ~ N
~--i
0
Calcd. for C~BH~~NaO,F~~0.3 Hz0
65.35 6.17 7.88
Found 65.38 6.02 8.05
The compoundB set forth in Examples 44 and 45 below were
prepared by procedures substantially as described above for Example 43.
EXAMPLE 44
(4S, 5S)-5-Cyclopropyl-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-
carboxylic acid~l-[4-(4-fluorophenyl)-4-hydroxy-cyclohexyl]-(3R)-
pyrrolidin-3-yl?amide hydrochloride
This compound was prepared from the product of Example 35,
Step D and the product of Example 43, Step C by a procedure similar to
that described in Example 25, Step E.
F
F
F ~ ~ O
HO N- 'N ..."
H ~
HCI O~O _ __
Analysis: Calculated for C~H~ZN~04F~ ~ HCL:
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C, 60.04; H, 5.74; N, 7.24
Found: C, 59.74; H, 5.76; N, ?.4b
Mass Spectrum: 544.3 (M+1), FAB
EXAMPLE 46
(45, 5S)-5-Cyclopropyl-4-(3,4-diffuorophenyl)-2-oxo-oxazolidine-3-
carboxylic acid(1-[4-(4-cyanophenyl)-cyclohexyl]-(3R)-pyrrolidin-3-
yl)amide
F
I \ F
NC ~ ~ O
N "N "",
H ~ Q
O?' O
Analysis: Calculated for C'QH9pN,09F2 ~ O.lb CHClg:
C, 65.54; H, b.87; N, 10.14
Found: C, 65.85; H, 5.80; N, 10.10
The compounds set forth in Examples 46-64 below were prepared
using procedures substantially the same as described in the preceding
Examples.
EXAMPLE 46
(4S)~4-(3,4-difluorophenyl)-2-ozo-ozazolidine-3-carboxylic acidll-[4-(4
cyanophenyi?-4-hydroxy-cyclohexyl]-(3ft)-pprrolidin-3-yl)amide
hydrochloride
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F
F
NC ~ \ N OII
HO N~N
H
HCI O~O
Analysis: Calculated for Cz~H2AN4O4F2 ~ HCL ~ 0.75 H20:
C, 57.85; H, 5.48; N, 10.00
Found: C, 57.88; H, 5.79; N, 9.97
EXAMPLE 47
(4S)-3-{1-[4-(4-Cyanophenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-4-
(3,4-difluorophenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-
carboxylic acid methyl ester
F
~ F
I~
\ O
N C N ~ COOMe
H I
O H CH3
Analysis: Calculated for C3,H33N5~4F2'
C, 64.45; H, 5.77; N, 12.12
Found: C, 64.04; H, 5.73; N, 11.82 _ -
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EXAMPLE 48
(45)-traps-4-(3,4-Difluorophenyl)-3-[1-{4-pyridin-2-yl-cyclohexyl)-(3R)
pyrrolidin-3-ylcarbamoyl]-6-methyl-2-oxo-1,2,3,4-tetrahydro-pyrimidine
5-carboxylic acid methyl ester
F
F
O
..~~N ~ C02Me
N I
N O~ N
H
To a solution of dry acetonitrile {20 mL) containing 666 mg (1.141 mmole)
of traps-4-(3,4-difluorophenyl)-3-[1-(4-pyridin-2-yl-cyclohexyl)-3R-
pyrrolidin-3-ylcarbamoyl]-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester (the preparation of which is
described in Example 9) was added 0.676 ml (5.33 mmole) of
chlorotrimethylsilane and 0.8 g (5.33 mmole) of sodium iodide. The
reaction vessel was fitted with an efficient reflux condenser and the
reaction mixture was heated to 65° C for 30 minutes. The reaction
mixture was subsequently heated to reflux for 30 minutes more. After
one hour, an additional 5.33 mmole each of chlorotrimethylsilane and
sodium iodide were added and then the same amount, once more, after
three hours. The reaction was terminated after six hours. All volatiles
were removed under reduced pressure and the residue was partitioned
between ethyl acetate (200 mL) and water. The organic phase was
washed with water and brine, then dried (sodium sulfate) and
concentrated to yield an amorphous solid. The crude product was
purified via preparative centrifugal chromatography (4 mm plate
thickness, 2-20% methanol/chloroform gradient); repurification via
preparative thick layer chromatography (1 mm plate thickness, 88:12:1.2
chloroform/methanol/conc. ammonium hydroxide elution, v/v) yielded
212 mg (34%) of the title compound in analytically pure form. This
material was converted to its salt form with HCl (gas) in ethyl acetate:
HPLC = >99% pure at 215 nm & 255 nm.
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NMR(CDC13, 400 MHz): Consistent with structure assignment and
confirms presence of solvent.
FAB MS: 554 (M+ + 1).
Analysis for C2gH33F2N504 ~ 2HCl ~ 0.7H20:
Calculated: C, 54.49; H, 5.74; N, 10.96.
Found: C, 54.46; H, 5.73; N, 10.64.
EXAMPLE 49
(4S)-3-{1 -[4-(2-Cyano-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3- ylcarbamoyl}-
4-(3,4-dilluoro-phenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydro-pyrimidine-5-
carboxylic acid methyl ester Hydrochloride.
F
H F /
N
C N H ~ ,~'H O
N ~ -OCH3
~HCI
O~N CH3
H
Analysis: Calcd. for C3,Ha;~F2N504~HCl~0.75H20
C, 59.32; H, 5.70; N, 11.16
Found: C, 59.08; H, 5.89; N, 10.87
EXAMPLE 50
(4S)-3-{1 -[4-(2-Cyano-4-fluoro-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl}-4-(3,4-difluoro-phenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydro-pyrimidine-5-carboxylic acid methyl ester Hydrochloride.
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F / I F
N O \ O
C N H .,. H
~HCI H ~ ~ OCH3
H
O~
Analysis: Calcd. for C32H34F3Nu~5~HCl~1.5H20
C, 55.77; H, 5.56; N, 10.16
Found: C, 55.52; H, 5.63; N, 10.02
EXAMPLE 51
(4S)-4-(3,4-Difluoro-phenyl)-3-(1 -[4-(4-fluoro-2-hydroxy-phenyl)-
cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6-methoxymethyl-2-oxo
1,2,3,4-tetrahydro-pyrimidine-5-carboxylic acid methyl ester
Hydrochloride.
F
F
/ H F /
N \
O H H ~ ,,.H O
N I ~OCH3
~HCI ~
O-' _ N
H O
Analysis: Calcd. for C3,H35F3N406~HC1~0.85H20~0.25C4H,o0_
C, 55.95; H, 5.90; N, 8.16
Found: C, 56.17; H, 5.51; N, 7.81
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EXAMPLE 52
(4S)-4-(3,4-Diffuoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid (1-[4 -(4
ffuoro-2-hydroxy-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3- yl}-amide
Hydrochloride.
F F
\ I H I \ F
I ~ 'N~ O
OH H II
N~N
~HCI H
O O
Analysis: Calcd. for C26H~pF3N304 ~ HC1 ~ 0.10H20
C, 57.64; H, 5.43; N, 7.76
Found: C, 57.42; H, 5.22; N, 7.88
EXAMPLE 53
(4S)-4-(3,4-Difluoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid (1-[4 -(2-
cyano-4-fluoro-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}-amide.
F / F
\ I I ~ F
_ N
CN H
H ~[
O~O
- _
Analysis: Calcd. for C2~H27F3N403~0.35H20
C, 62.50; H, 5.38; N, 10.80
Found: C, 62.87; H, 5.53; N, 10.42
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EXAMPLE 54
(4S)-4-(3,4-Diffuoro-phenyl)-2-oxo-oxazolidine-3-carboxylic acid {1-[4 -(2
cyano-4-fluoro-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}-amide
Hydrochloride.
F / F
~ I ~ ~ F
i a _ N
CN H
H N
~HCI
O
Analysis: Calcd. for CZ7H27F3N403~HCl
C, 59.07; H, 5.14; N, 10.21
Found: C, 59.38; H, 5.42; N, 9.87
EXAMPLE 55
(4S,5S)-4-(3,4-Difluoro-phenyl)-5-methyl-2-oxo-oxazolidine-3-carboxylic
acid (1-[4 -(2-cyano-4-fluoro-phenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl}-
amide Hydrochloride.
F /
I ~HCI F
C N ~N ~ \ F
NJ ,."~CH3 . _
O~O
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Analysis: Calcd. for C28H29F3N4O3~HC1~1.25H20
C, 57.43; H, 5.59; N, 9.57
Found: C, 57.19; H, 5.68; N, 9.79
EXAMPLE 56
(4S,5S)-5-Cyclopropyl-4-(3,4-difluoro-phenyl)-2-oxo-oxazolidine-3-
carboxylic acid {1-[4 -(2-cyano-4-fluoro-phenyl)-cyclohexyl]-(3R)-
pyrrolidin-3y1}-amide Hydrochloride.
F /
~HCI F
C N ~N ~ \ F
H N
", a
0
Analysis: Calcd. for C3~ H3,F3N40;~~HCl~0.55H20
C, 60.15; H, 5.57; N, 9.35
Found: C, 59.85; H, 5.77; N, 9.14
EXAMPLE 57
(4S)-4-(3,4-Difluoro-phenyl)-3-{1-[4-(2-fluoro-phenyl)-4-hydroxy-cyclohex-
1-yl]-(3R)-pyrrolidin-3-ylcarbamoyl}-6-methyl-2-oxo-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester hydrochloride
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F
F--~~ ~~ o~ i
H
~~ N \
N ~N
~HCI O ~H
HO
F
Analysis: Calcd. for C3~H3;~F3N405~HCl~0.60 H20~0.35 EtOAc
C,56.73; H,5.76; N, 8.43
Found: C, 56.73; H,5.83; N, 8.45
EXAMPLE 58
(4S)-trans-4-(3,4-Difluoro-phenyl)-3-(1-[4-(2-fluoro-phenyl)-cyclohexyl]-
(3R)-pyrrolidin-3-ylcarbamoyl}-6-methyl-2-oxo-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester hydrochloride
F
F ~ ~ O
O
H
~~ N
N ~N
~HCI O O H
w _
F
Analysis: Calcd. for C3oH32F3N404~HC1~0.85 H20~0.10 EtOAc
C, 57.94; H, 5.68; N, 8.89 w -
Found: C, 57.93; H,5.91; N, 8.94
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EXAMPLE 59
(4S)-cis-4-(3,4-Difluoro-phenyl) -3-[1-(4-hydroxy-4-pyridin-2-yl-cyclohexyl)
(3R)-pyrrolidin-3-ylcarbamoyl] -2-oxo-1,2,3,4-tetrahydro-pyrimidine-5
carboxylic acid methyl ester hydrochloride
F
F \ / Y-O
(~ N N
.N ~ N
O
HO '2HC1 O H
,N
Analysis: Calcd. for C28H3,FZN50~~2HC1~1.15 Hz0
C,51.80; H,5.4; N,10.79
Found: C, 51.79; H,5.60; N, 10.86
EXAMPLE 60
(4S)-tr~ns-4-(3,4-Difluorophenyl)-3-[1-(4-pyridin-2-yl-cyclohexyl)-(3R)
pyrrolidin-3-ylcarbamoyl]-2-oxo-1,2,3,4-tetrahydro-pyrimidine-5
carboxylic acid methyl ester
F,
F \ / OY-O
N
N
.N
O ~N
O H
/N _ _
Analysis: Calcd. for CZ8H3,FZN504~0.75 Hz0
C,60.80; H,5.92; N,12.66
Found: C, 60.74; H,5.72; N,13.05
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E~;AMPLE 61
(4S)-traps-3{ 1-[4-(2-Cyano-phenyl)-piperidin-1-yl]-(3R)-pyrrolidin-3-
ylcarbamoly}-4-(3,4-difluoro-phenyl)-2-oxo-1,2,3,4-tetrahydro-pyrimidine-
5-carboxylic acid methyl ester
F
F-~~
N
N ~ ~N
.,,. O H
O
w
~ CN
Analysis: Calcd. for C3oH3,F2N504 '0.75 H20
C, 62.43; H, 5.68; N, 12.14
Found: C, 62.43; H, 5.45; N, 12.27
EXAMPLE 62
(4S,5S)-traps-5-Cyclopropyl-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-3-
carboxylic acid { 1-[ 4-(4-fluorophenyl)-cyclohexyl]-(3R)-pyrrolidin-3-yl-
amide
F
~ ,..,. N O ~ F
F -
N
H
O ~.""
O
Calcd. for C29H32N3~3F3 ~ 0~35 H2O' O.1 CHZC12
C, 64.44; H, 6.11; N, 7.75
Found C, 64.39; H, 5.97; N, 7.77
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EXAMPLE 63
(4S)-4-(3,4-Difluoro-phenyl) -3-[1-(4-hydroxy-4-pyridin-2-yl-cyclohexyl)-
(3R)-pyrrolidin-3-ylcarbamoyl]-6-methyl -2-oxo-1,2,3,4-tetrahydro-
pyrimidine-5-carboxylic acid methyl ester hydrochloride
F
HO O / ~ F
N~ ~
-N H N
O~ / C02Me
~ 2 HCI N
H Me
Analysis: Calcd. for CZ9H33F2N5O56 ~ 2 HCl
C, 50.26; H, 5.91; N,10.11
Found: C, 50.24; H, 5.72; N, 9.82
EXAMPLE 64
(4S)-trans-4-(3,4-Difluorophenyl)-5-methyl-2-oxooxazolidine-3-carboxylic
acid-[1-(4-pyridin-2-yl-cyclohexyl)-(3R)-pyrrolidin-3-yl]amide
hydrochloride
F
F
/ ~ "", N O /
_N~ ~N
H N
"'Me
O O ~ 2 HCl
Mass Spectrum (FAB) Calcd. for C26H27F2N4O3
Found: 485 (M+1)
EXAMPLE 65
trans-4S-( 3,4-Difluorophenyl )-6-methyl-2-oxo-3- { 1-[4-( 1-oxopyridin-2-yl )-
cyclohexyl]-3R-pyrrolidin-3-ylcarbamoyl?-1,2,3,4-tetrahydropyrimidine-5-
carboxylic acid methyl ester hydrochloride
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F
F
H O
~~,, N--~N C 02M a
o N~ I
O~N Me
H
Step A: 2-(1,4-Dioxaspiro[4.5]dec-7-en-8-yl)pyridine
To a solution of trifluoromethanesulfonic acid 1,4-dioxa-
spiro[4.5]dec-7-en-8-yl ester (25.41 g, 88.15 mmol) in THF (266 mL) at
room temperature was added palladium tetrakistriphenylphosphine
(5.08 g, 4.4 mmol) under an inert atmosphere. To this mixture was
added a solution of 2-pyridyl zinc bromide (264.4 mL of a 0.5 N solution
in THF, 132.2 mmol) resulting in a slight exotherm. The resulting
reaction mixture was then stirred 15 minutes. The reaction was
quenched with the addition of 200 mL of saturated sodium bicarbonate
solution. After stirring 5 minutes more, the phases were separated and
the aqueous layer was extracted with ethyl acetate The combined
organic extracts were washed with saturated sodium chloride, dried
with sodium sulfate, and concentrated in vacuo. The crude material
was chromatographed over silica gel eluting with 25% ethyl
acetate/hexane to give 14.6 (76%) of homogeneous product.
Step B: 2-(1,4-Dioxaspiro[4.5]dec-8-yl)pyridine
To a solution of 14.6 g (67.19 mmol) of 2-(1,4-
dioxaspiro[4.5]dec-7-en-8-yl)pyridine in ethyl acetate (400 mL) was added
4.6 g of 10% palladium on carbon catalyst. The resulting suspension was
then hydrogenated at ambient temperature and atmospheric pressure
until all the starting material was consumed ( 10.5 hr). The reaction
mixture was filtered through celite and the filtrate was concentrated to
give the product as an oily solid.
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Step C: 2-(1,4-Dioxaspiro[4.5]dec-8-yl)pyridine N-oxide
To an ice cold solution of 3 g (13.68 mmol) of 2-(1,4-
dioxaspiro[4.5]dec-8-yl)pyridine in chloroform (46 mL) was added 4.51 g
(14.36 mmol) of meta-chloroperoxybenzoic acid (50-60%). The reaction
mixture was stirred and allowed to come to room temperature over a
four hour period. The volume of the reaction mixture was reduced by
approximately 75% under reduced pressure and the residual material
was partitioned between ethyl acetate and 1 N aqueous sodium hydroxide
solution. The organic phase was separated and washed with water and
brine, then dried (sodium sulfate) and concentrated to yield 3 g of the
product as an oil.
Step D: 2-(4-Oxocyclohexyl)pyridine N-oxide
2-(1,4-Dioxaspiro[4.5]dec-8-yl)pyridine N-oxide (3 g, 12.75
mmol) was combined with glacial acetic acid (20 mL), concentrated
hydrochloric acid (10 mL), and water (10 mL). The resulting mixture
was heated to 45°C for 24 hours. The reaction was cooled to 0°C
and the
pH was adjusted to 6 with 20% sodium hydroxide solution. The phases
were separated and the aqueous layer was extracted with ethyl acetate (3
X 100 mL). The combined organic extracts were washed with water and
brine, then dried (sodium sulfate), and concentrated under reduced
pressure to yield 1.8 g of the product as an oil.
Step E: 2-[trans-4-(3R-3-Aminopyrrolidin-1-yl)-cyclohexyl]pyridine
N-oxide
To a solution of 17.8 mL of methanol containing 2-(4-
oxocyclohexyl)pyridine N-oxide (1.8 g, 9.41 mmol) and 1.75 g (9.41 mmol)
of (3R)-(+)-3-(tert-butoxycarbonylamino)-pyrrolidine was added 3.23 rnL
(56.4 mmol) of glacial acetic acid under an inert atmosphere. The
resulting solution was stirred at ambient temperature for 20 minutes,
cooled to 0°C, and treated with four equal portions of solid sodium
cyanoborohyride (total: 296 mg, 4.71 mmol). The reaction mixture was
stirred at 0°C for 20 minutes more. All volatiles were removed under
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reduced pressure and the residue was taken up in 200 mL of ethyl
acetate. The organic layer was washed with 10% sodium carbonate
solution and brine, then dried (sodium sulfate) and concentrated to yield
2.86 g of the crude product as an oil. The desired trans-2-[4-(3-tert-
butoxycarbonylaminopyrrolidin-1-yl)-cyclohexyl]-pyridine N-oxide was
obtained via preparative centrifugal chromatography on silica gel
(chloroform-methanol gradient elution). A portion of this material (544
mg, 1.505 mmol) was dissolved in 300 mL ethyl acetate at 0°C, and HCl
gas was bubbled through the solution for 10 min. The reaction mixture
was allowed to come to room temperature over a 1 hour period. All
volatiles were rotoevaporated under reduced pressure to give a white
solid. This material was resuspended in 200 mL of ethyl acetate and
treated with 10 ml of 10% sodium carbonate solution. After 10 minutes
the phases were separated and the organic layer was dried (sodium
sulfate). The aqueous layer was concentrated to dryness and the residue
was stirred with a chloroform-methanol mixture (4:1, v/v). The
suspension was filtered and the filtrate was combined with the ethyl
acetate extracts. The combined organic extracts were then concentrated
and azeotropicaily dried with toluene to give 400 mg of the title compound
as a tan powder.
Step F: trczns-4S-(3,4-Difluorophenyl)-6-methyl-2-oxo-3-(1-[4-(1-
oxopyridin-2-yl)-cyclohexyl]-3R-pyrrolidin-3-ylcarbamoyl)-
1,2,3,4-tetrahydropyrimidine-5-carboxylic Acid Methyl Ester
Hydrochloride
2-[trans-4-(3R-3-Aminopyrrolidin-1-yl)-cyclohexyl]pyridine
N-oxide (300 mg, 1.148 mmol) and (+)-3-(4-nitrophenoxycarbonyl)-4S-(3,4-
difluorophenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylic
acid methyl ester (514 mg, 1.148 mmol) were combined in 20 mL of dry
tetrahydrofuran at room temperature under nitrogen. The solution was
stirred for 10 minutes, an additional 50 mg of (+)-3-(4- - --
nitrophenoxycarbonyl)-4S-(3,4-difluorophenyl)-6-methyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester was added, and
stirring was continued for 1 hour. The reaction mixture was
concentrated in vacuo and the residue was purified directly via
preparative centrifugal chromatography on silica gel (first ethyl acetate
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and then chloroform-methanol gradient elution: CHC13/MeOH, 100%/0%
to 85%/15%) to give the title compound as the free base. The
hydrochloride salt was prepared by dissolving the chromatographed
product in ice cold ethyl acetate and treating this solution with a solution
of ethyl acetate saturated with HCl gas. In this way, the title compound
was obtained analytically pure as an off white solid: m.p. 195-200°C.
Anal. Calcd. for C29HssF2NsOs'HCl~2.2H20: C, 53.78; H,
5.60; N, 10.59.
Found: C, 53.94; H, 5.60; N, 10.59%.
EXAMPLE 66
tracns-2-( 3,4-Difluorophenyl )-1-[ 1-(4-pyridin-2-yl-cyclohexyl )-3R-
pyrrolidin-3-ylcarbamoyl]-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydro-
pyrimidine
O
f~
H
O
H
The title compound was prepared in accordance with the
following scheme:
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F
NaOH I /
MeOH
54-71% H~
G ~. O~ O N
H
17 18 _ 19
2.OM LDA
,- 02N
02 ~ o
/ O
0
20 F
F
/ \ / _N~ O
N NH2 N N~N
H
21 O"N
H
Step A: To a solution of (+)-DHP 17 (4.63 g, 14.7 mmol) in a methanol
(100 ml) was added sodium hydroxide (2.94 g, 73.6 mmol). The resulting
mixture was refluxed at 90 oC for 16 hours. After cooling to room
temperature the solvent was removed in vaccuo. The solid was dissolved
in CHZC12 and H20 then neutralized with 10% aqueous HCl solution.
The organic layer was dried over Na2S04, concentrated, and purified by
PCTLC (7% MeOH in CHC13 with 2% NH40H) to afford a 2.65 g mixture
of 18 and 19 (71°lo yield). The 'H NMR was consistent with the assigned
structure.
MS (FAB) 255 (M+1)
Step B (alternative to Step A): To a solution of (+)-DHP 17 (5.36 g, 17.0
mmol) in a methanol (150 ml) was added 1N NaOH (10 ml). The
resulting mixture was refluxed at 90 oC for 16 hours. After cooling to
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room temperature the solvent was removed in uacuo. The solid was
dissolved in CH2C1~ and H20 then neutralized with 10% aqueous HCl
solution. The organic layer was dried over NalS04, concentrated, and
purified by PCTLC (?% MeOH in CHC13 with 2% NH40H) to afford a 2.35
g mixture of 18 and 19 (54% yield). The 'H NMR is consistent with the
assigned structure.
MS (FAB) 255 (M+1)
Step C: The title compound was prepared by reacting a mixture of
18 and 19 (1.93 g, 7.59 mmol) with 4-nitrophenoxycarbonyl chloride (1.5
equivalents) in LDA (1.1 equivalents) at -78°C until the reaction was
completed as determined by GLC. 0.488 g (15°!o yield) of 20 was
obtained.
The 'H NMR was consistent with the assigned structure.
Step D: The title compound was obtained from 20 (0.119 g, 0.284
mmol) using the procedure described for step F in Example 1. The 'H
NMR was consistent with the assigned structure.
C28H33FZN~O3 MS (FAB) 526 (M+H).
EXAMPLE 67
(4S,5R)-trans-4-(3,4-Difluorophenyl-3-( 1-[4-(4-fluoro-2-
methoxycarbonylphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-2-
oxo-oxazolidine-5-carboxylic acid methyl ester
F
F
F ~ ~~~N~ O N,C02Me
C02Me H~ O
O
Step A: trans-3,4-Difluorocinnamic acid methyl ester
To a solution of trans-3,4-difluorocinnamic acid (10 g, 54 mmol) in
300 mL methanol was added concentrated sulfuric acid (2 mL). The
solution was stirred 48 h at ambient temperature and then concentrated
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in vacuo. The residue was taken up in ethyl acetate (500 mL) and
washed with saturated sodium bicarbonate (2 x 100 mL), brine ( 1 x 100
mL), dried with magnesium sulfate, and concentrated in vacuo to
provide tracns-3,4-difluorocinnamic acid methyl ester (10.7 g, 54 mmol,
100%) as a white solid.
1H NMR dH (400 MHz, CDC13) 7.59 (d, 1H, J = 15.9), 7.34 (m, 1H),
7.24 (m, 1H) 7.18 (dd, 1H, J = 9.9, 2.0), 6.35 (d, 1H, J = 16.1), 3.81 (s,
3H).
Step B: (2R, 3S)-N-Benzyloxycarbonyl-3-amino-3-(3,4-
difluorophenyl)-2-hydroxypropionic acid methyl ester
A solution of NaOH (4.1 g, 103 mmol) was prepared in 175 mL
water. Potassium osmate dihydrate (491 mg, 1.3 mmol) was dissolved in
35 mL of this NaOH solution, resulting in a dark pink homogeneous
mixture. To a 1000 mL round bottom flask is added the remaining
NaOH solution prepared above, 135 mL n-propanol and benzyl cabamate
(9.8 g, 110 mmol). The suspension was stirred at ambient temperature
for 30 min wherein the mixture was nearly homogeneous. The reaction
flask was placed in a room temperature water bath and the surrounding
lights were turned off. Freshly prepared t-butylhypochlorite (11.2 mL,
103 mmol) was added dropwise with vigorous stirring, and the reaction
stirred an additional 15 min. In a separate 250 mL round bottom flask
was suspended trans-3,4-difluorocinnamic acid methyl ester (6.6 g, 33.3
mmol) and (DHQ)ZPHAL (1.3 g, 1.7 mmol) in 100 mL n-propanol. The
suspension was added to the above reaction mixture and the residue
rinsed into the reaction flask (2 x 10 mL). To the reaction was added the
above prepared solution of potassium osmate dihydrate. The resulting
green solution became amber/brown over 1 h. Sodium metabisulfite (66
g, 347 mmol) was added and the resulting suspension stirred 3 h when it
was poured into a separatory funnel containing ethyl acetate (200 mL)
and the layers separated. The aqueous layer was extracted with ethyl
acetate ( 150 mL) and the combined organics washed with brine ( 100 ~mL),
dried with magnesium sulfate, and concentrated in vacuo to provide a
pale yellow solid. The crude material was passed through silica (25°l0
ethyl acetate/hexane) to give (2R, 3S)-N-benzyloxycarbonyl-3-amino-3-
(3,4-difluorophenyl)-2-hydroxypropionic acid methyl ester contaminated
with benzyl carbamate.
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Step C: (2R, 3S)-3-Amino-3-(3,4-difluorophenyl)-2-hydroxypropionic
acid methyl ester
(2R, 3S)-N-benzyloxycarbonyl-3-amino-3-(3,4-difluorophenyl)-2-
hydroxypropionic acid methyl ester (>12.2 g, 33.3 mmol maximum) was
dissolved in 750 mL ethanol. The flask was purged and filled with argon
three times. Palladium on carbon (2 g, 10% wt) was added under argon
and the suspension was again purged and filled with argon three times.
The suspension was then purged, filled with hydrogen, and stirred 16 h.
The suspension was purged, filled with argon three times, filtered
through celite and concentrated in vacuo to give (2R, 3S)-3-amino-3-(3,4-
difluorophenyl)-2-hydroxypropionic acid methyl ester (5.8 g, 25 mmol,
75% from trans-3,4-difluorocinnamic acid methyl ester).
'H NMR dH (400 MHz, CDC13) 7.26 (m, 1H), 7.15-7.08 (m, 2H), 4.28
(s, 2H), 3.82 (s, 3H), 2.48 (bs, 2H).
Step D: (4S,5R)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-5-carboxylic acid
methyl ester
To a solution of (2R, 3S)-3-amino-3-(3,4-difluorophenyl)-2-
hydroxypropionic acid methyl ester (5.8 g, 25 mmol) in 250 mL
tetrahydrofuran at 0°C was added N,N-diisopropylethylamine (8.75 mL,
50 mmol) and triphosgene (2.48 g, 8.4 mmol). The reaction was stirred
at 0°C for 30 min when it was poured over ethyl acetate (200 mL) and
saturated sodium carbonate solution ( 100 mL). The layers were
separated, the organic layer washed with saturated sodium carbonate
solution (1 x 100 mL), dried with magnesium sulfate, and concentrated
in vacuo to provide a pale yellow oil. The material was triturated with
25% ethyl acetate/hexane from dichloromethane to provide (4S,5R)-4-
(3,4-difluorophenyl)-2-oxo-oxazolidine-5-carboxylic acid methyl ester.
The recovered mother liqour was passed through silica (50% ethyl - -
acetate/hexane) to give an additional 1.1 g (4.8 g total, 18 mmol, 75%)
'H NMR dH (400 MHz, CDC13) 7.25-7.20 (m, 2H), 7.15 (m, 1H), 6.33
(bs, 1H), 4.98 (d, 1H, J = 5.1), 4.72 (d, 1H, J = 5.3), 3.89 (s, 3H).
FABMS M+H = 258
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Step E: (4S,5R)-4-(3-,4-difluorophenyl)-2-oxo-oxazolidine-3,5-
dicarboxylic acid methyl ester 3-(4-nitrophenyl) ester
To a solution of (4S,5R)-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-5-
carboxylic acid methyl ester (910 mg, 3.5 mmol) in anhydrous
tetrahydrofuran (50 mL), cooled to -78°C under argon, was added a THF
solution of lithium bis(trirnethylsilyl)amide (3.5 mL, 3.5 mmol)
dropwise. The reaction mixture was warmed to 0°C in an ice bath,
stirred 30 minutes, then returned to -78°C. In a separate flask, p-
nitrophenylchloroformate (714 mg, 3.54 mmol) was dissolved in
anhydrous tetrahydrofuran (40 mL) under argon and cooled to -78°C.
The above prepared anion solution was added via cannula to the
chloroformate solution and the reaction mixture was stirred 1 h at -
78°C. The reaction mixture was treated with ethyl acetate (150 mL) and
the resulting solution was washed with water ( 1 x 150 ml) , brine ( 1 x
150 ml) and dried over magnesium sulfate and filtered. The volitiles
were removed under reduced pressure and the resulting oil was
triturated with diethyl ether. Ether was twice decanted from the
resulting pale yellow solid to give (4S,5R)-4-(3-,4-difluorophenyl)-2-oxo-
oxazolidine-3,5-dicarboxylic acid methyl ester 3-(4-nitrophenyl) ester (1.3
g, 3.1 mmol, 87%).
FAB MS: m/z=423 (M+H)
Step F: (4S,5R)-trans-4-(3,4-Difluorophenyl-3-(1-[4-(4-fluoro-2-
methoxycarbonylphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl)-2-oxo-oxazolidine-5-carboxylic acid methyl
ester
To a solution of trans-2-[4-(3-amino-(3R)-pyrrolidin-1-
yl)cyclohexyl]-5-fluorobenzoic acid methyl ester (145 mg, 0.37 mmol) in
dry, degassed N,N-dimethylformamide (3 mL) was added N,N-
diisopropylethylamine (130 ~L, 0.74 mmol) followed (4S,5R)-4-(3-,4- - --
difluorophenyl)-2-oxo-oxazolidine-3,5-dicarboxylic acid methyl ester 3-(4-
nitrophenyl) ester (156 mg, 0.37 mmol). The reaction mixture was
stirred at ambient temperature for 4 h. The volatiles were removed
under reduced pressure and residue dissolved in ethyl acetate, washed
with 10% aqueous sodium carbonate solution (8 x 100 ml), brine (1 x 100
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ml), dried over magnesium sulfate and filtered. The volatiles were
removed under reduced pressure and the resulting oil was purified by
pressurized silica gel chromatography (2-5% methanol in ethyl acetate)
to afford (4S,5R)-trans-4-(3,4-difluorophenyl-3-{1-[4-(4-fluoro-2-
methoxycarbonylphenyl)-cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl}-2-
oxo-oxazolidine-5-carboxylic acid methyl ester as a white foamy solid.
The hydrochloride salt was prepared according to standard procedures
and isolated as a white solid (125 mg, 0.2 mmol, 56%).
'H NMR dH (400 MHz, CD30D) 7.46 (m, 2H), 7.33 (m, 2H), 7.24 (m,
2H), 5.54 (dd, J= 3.85, 1.83 Hz, 1H), 5.03 (dd, J= 3.84, 2.01 Hz, 1H), 3.89
(s,
3H), 3.87 (s,3H), 3.82-3.20 (m, 8H), 2.57 (m, 1H), 2.30 (m, 3H), 1.65 (m,
4H).
Analysis: Calcd for C30 H32 N3 07 F3 ~HCl ~ 0.15 Et20 ~0.85 H20,
C 55.14, H 5.47, N 6.31.
Found: C 55.13, H 5.44, N 6.16
HPLC retention time = 9.082 min, purity= 96%
FAB MS: m/z=604 (M+H)
The compounds in Examples 68-73 were prepared by procedures
substantially as described above for Example 67, Step F.
EXAMPLE 68
(45,5R)-traps-4-(3,4-Difluorophenyl)-3-(1-{4-[4-fluoro-2-(2,2,2-
trifluoroethoxy)phenyl]cyclohexyl}-(3R)-pyrrolidin-3-ylcarbamoyl)-2-oxo-
oxazolidine-5-carboxylic acid methyl ester
F
F
F ~ ~~~N~ O ,~~C02Me _ __
U
OCH2CF3 N ~O
H
O
FAB MS: m/z=644 (M+H)
Analysis: Calcd for C30H31N3O6F6~F3CC02H,
C 50.73, H 4.26, N 5.55.
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Found: C 50.34, H 4.13, N 5.50.
EXAMPLE 69
(4S,5R)-tracns-4-(3,4-Difluorophenyl )-2-oxo-3-[ 1-(4-pyridin-2-ylcyclohexyl)-
(3R)-pyrrolidin-3-ylcarbamoyl]-oxazolidine-5-carboxylic acid methyl ester
F
F
~~~N~ O ~~~CO2M a
N
H ~O
,/O
FAB MS: m/z=529 (M+H)
Analysis: Calcd for C27H30N4O5F2~2HC1~0.95H20~0.3EtOAc
C 52.85, H 5.80, N 8.55
Found: C 52.86, H 5.45, N 8.55
EXAMPLE 70
(4S,5R)-trczns-4-(3,4-Difluorophenyl )-3-( 1-[4-(4-fluoro-2-
methoxyphenyl)cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl]-2-oxo-
oxazolidine-5-carboxylic acid methyl ester
F
F
F ~ ~~~N~
OMe N
H
FAB MS: m/z=576 (M+H)
Analysis: Calcd for C29H32N3O6F3~HCl~0.25H20
C 56.49, H 5.48, N 6.82.
Found: C 56.52, H 5.50, N 7.03. - -
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EXAMPLE 71
(4S,5R)-3-{ 1-[4-Cyano-4-(2-methoxyphenyl)cyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl}-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-5-carboxylic acid
methyl ester
F
N F w
NI ~ ~,,C 02M a
O M a H N~O
'IO
FAB MS: m/z=583 (M+H)
Analysis: Calcd for C30H32N406F2~F3CC02H~0.6CH2C12
C 52.37, H 4.61, N 7.49.
Found: C 52.32, H 4.37, N 7.24.
EXAMPLE 72
(4S,5R)-3-{ 1-[4-Cyano-4-(2-fluorophenyl)cyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl}-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-5-carboxylic acid
methyl ester
F
F
- NC
N~ O ~~,C02Me
F H~ O
O
FAB MS: m/z=571 (M+H)
Analysis: Calcd for C29H29N405F3~HCl~0.65H20
C 56.30, H 5.10, N 9.06.
Found: C 56.33, H 5.22, N 8.83.
EXAMPLE 73
(4S,5R)-trans-3-{1-[4-(2-Cyanophenyl)cyclohexyl]-(3R)-pyrrolidin-3-
ylcarbamoyl}-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-5-carboxylic acid
methyl ester
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F
F
~~~N~ O ~C02Me
N
C N H ~O
IlO
FAB MS: m/z=553 (M+H)
Analysis: Calcd for C29H30N405F2 ~HCl ~ 0.5H20
C58.24,H5.39,N9.37.
Found: C 58.23, H 5.42, N 9.07.
EXAMPLE 74
(4S,5R)-trans-4-(3,4-difluorophenyl)-5-hydroxymethyl-2-oxo-oxazolidine-
3-carboxylic acid {1-[4-(4-fluorophenyl)cyclohexyl]-(3R)-pyrrolidin-3-yl}
amide
F
F
F \ m ~~ H
N
H
Step A: (4S,5R) 4-(3,4-Difluorophenyl)-5-hydroxymethyl-oxazolidin-
2-one
To a solution of (4S,5R)-4-(3,4-difluorophenyl)-2-oxo-oxazolidine-5-
carboxylic acid methyl ester [(200 mg, 0.8 mmol) product of Example 67,
Step D] in tetrahydrofuran (10 mL) at 0°C was added a 2 M solution
of
lithium borohydride in tetrahydrofuran (0.4 mL, 0.8 mmol). After
stirring for 20 min at 0°C, saturated sodium bicarbonate (20 mL) was
added and the mixture stirred at ambient temperature for 20 min. Ethyl
acetate (50 mL) was added and the layers separated. The organic layer
was washed with brine (1 x 10 mL), dried with magnesium sulfate,
filtered and concentrated in vacuo to provide (4S,5R) 4-(3,4-
difluorophenyl)-5-hydroxymethyl-oxazolidin-2-one as a white solid (180
mg, 0.8 mmol, 100%)
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'H NMR dH (400 MHz, CDCh) ?.23-7.15 (m, 2H), 7.10-7.07 (m, 1H),
6.48 (bs, 1H), 4.89 (d, 1H, J = 6.8), 4.31 (dt, IH, J = 6.6, 2.9), 3.96 (dd,
1H, J
= 12.82, 2.75), 3.70 (bdd, 1H, J =12.1, 2.2), 3.53 (be, 1H).
Step B: (45,5R) 4-(3,4-Difluorophenyl)~-(tetrahydropyran-2-
yloxymethyl)-oxazolidin-2-one
To a solution of (4S,5R) 4-(3,4-difluotophenyl)-5-hydroxymethyl-
oxazolidin-2-one (695 mg, 3.0 mmol) in dry dichloromethane (30 mL)
was added 2,3-dihydropyran (0.3 mL, 3.6 mmol) and camphorsulfonic
acid (70 mg, 0.3 mmol). The reaction mixture was stirred at ambient
temperature for 3 h. The reaction mixture was diluted with
dichloromethane (100 mL) and washed with saturated sodium
bicarbonate solution (2 x 100 ml), brine (1 x 100 ml), dried over
magnesium sulfate and filtered. The volatiles were removed under
reduced pressure and the resulting solid was purified by pressurized
silica gel chromatography (I:1 then 2:1 ethyl acetate:hexane) to afford
(4S,5R)-4-(3,4-difluorophenyl)-5-(tetrahydropyran-2-yloxymethyl)-
oxazolidin-2-one as a colorless oil (750 mg, 2.4 mmol, 800).
FAB MS: m/z-314 (M+H' )
Step C: (4S,5R)-4-(3,4-Difluorophenyl)_2-oxo-5-(tetrahydropyran-2-
yloxymethyl)-oxazolidine-3-carboxylic acid 4-nitrophenyl
ester
To a solution of (4S,5R)-4-(3,4-difluorophenyl)-5-(tetrahydropyran-
2-yloxymethyl)-oxazolidin-2-one (910 mg, 2.9 mmol) in anhydrous
tetrahydrofuran (60 mL) cooled to -78°C under argon, was added a TAF
solution of lithium bis(trimethylsilyl)amide (2.9 mL, 2.9 miuol)
dropwise. The reaction mixture was warmed to 0°C in an ice bath and
stirred 45 min, and then returned to -~8°C. Meanwhile, in a separate
dried flask, the p-nitrophenylchloroformate (586 mg, 2.9 mmol) was
dissolved in anhydrous tetrahydrofuran (50 mL) under argon and cooled
to -78°C. Then the above prepared anion solution was added via cannula
to the chloroformate solution and reaction mixture was stirred 1 h at -
78°C. The reaction mixture was treated with ethyl acetate (1b0 mL). The
resulting solution was washed with water (1 x 150 ml), brine (1 x 150
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mL), dried over magnesium sulfate and filtered. The volatiles were
removed under reduced pressure to give (4S,5R)-4-(3,4-difluorophenyl)-2-
oxo-5-(tetrahydropyran-2-yloxymethyl)-oxazolidine-3-carboxylic acid 4-
nitrophenyl ester as a yellow foam (1.3g, 2.8 mmol, 96%).
FAB MS: m/z=479 (M+H+)
Step D: (4S,5R)-traps-4-(3,4-Difluorophenyl)-5-(tetrahydropyran-2-
yloxymethyl-2-oxo-oxazolidine-3-carboxylic acid { 1-[4-(4-
fluorophenyl)cyclohexyl]-(3R)-pyrrolidin-3-yl} amide
To a solution of trcxns-1-[4-(4-fluorophenyl)cyclohexyl]-(3R)-
pyrrolidin-3-ylamine dihydrochloride (210 mg, 0.63 mmol) in dry
tetrahydrofuran (4 mL) was added N,N-diisopropylethylamine (219 ~tL,
1.26 mmol) followed by (4S,5R)-4-(3,4-difluorophenyl)-2-oxo-5-
(tetrahydropyran-2-yloxymethyl)-oxazolidine-3-carboxylic acid 4-
nitrophenyl ester (300 mg, 0.63 mmol). The reaction mixture was stirred
at ambient temperature for 18 h when the volatiles were removed under
reduced pressure and residue dissolved in ethyl acetate (100 mL) and
washed with 10% aqueous sodium carbonate solution (8 x 100 ml), brine
(1 x 100 ml), dried over magnesium sulfate and filtered. The volatiles
were removed under reduced pressure and the resulting oil was purified
by pressurized silica gel chromatography (3:1 then 1:0 ethyl
acetate:hexane to afford (4S,5R)-traps-4-(3,4-difluorophenyl)-5-
(tetrahydropyran-2-yloxymethyl-2-oxo-oxazolidine-3-carboxylic acid {1-[4-
(4-fluorophenyl)cyclohexyl]-(3R)-pyrrolidin-3-yl} amide
as a white foam (153 mg, 0.25 mmol, 40%).
Step E: (4S,5R)-traps-4-(3,4-Difluorophenyl)-5-hydroxymethyl-2-oxo-
oxazolidine-3-carboxylic acid {1-[4-(4-
fluorophenyl)cyclohexyl]-(3R)-pyrrolidin-3-yl} amide
To a solution of (4S,5R)-traps-4-(3,4-difluorophenyl)-5-
(tetrahydropyran-2-yloxymethyl-2-oxo-oxazolidine-3-carboxylic acid { 1-[4-
(4-fluorophenyl)cyclohexyl]-(3R)-pyrrolidin-3-yl} amide (153 mg, 0.26
mmol) was dissolved in methanol (5 mL) and p-toluenesulfonic acid (50
mg, 0.26 mmol) was added. The reaction mixture was stirred at ambient
temperature for 17 h. The volatiles were removed under reduced
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pressure and the residue was taken up in ethlyl acetate (100 mL) and
washed with saturated sodium carbonate solution (3 x 100 ml), brine (1 x
100 ml), dried over magnesium sulfate and filtered. The volatiles were
removed under reduced pressure to afford an oil which was lyophilized
at reduced pressure from acetonitrile and HCl (1N aqueous solution) to
afford (4S,5R)-trans-4-(3,4-difluorophenyl)-5-hydroxymethyl-2-oxo-
oxazolidine-3-carboxylic acid {1-[4-(4-fluorophenyl)cyclohexyl]-(3R)-
pyrrolidin-3-yl} amide.
HPLC: retention time= 8.05 min, purity=94%
FAB MS: m/z=571 (M+H)
Analysis: Calcd for C27 H30 N3 04 F3 ~HCl ~ 0.9 HZO
C 56.87, H 5.80, N 7.37.
Found: C 56.68, H 5.40, N 7.66.
The compound of Example 75 was prepared by procedures
substantially as described above for Example 74, Steps E and F.
EXAMPLE 75
(4S,5R)-trans-4-(3,4-Difluorophenyl)-5-hydroxymethyl-2-oxo-oxazolidine-
3-carboxylic acid {1-[4-(4-fluoro-2-methoxyphenyl)cyclohexyl]-(3R)-
pyrrolidin-3-yl} amide
F
F
F \ ~ ~ ~~ H
N
OMe H
FAB MS: m/z=548 (M+H)
Analysis: Calcd for C28H32N305F3~HCl~0.7H20
C 56.36, H 5.81, N 7.04.
Found: C 56.35, H 5.82, N 6.76.
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EXAMPLE 76
(4S,5R)-trczns-4-(3,4-Difluorophenyl)-3-{ 1-[4-(4-fluoro-2-
methoxyphenyl)cyclohexyl]-(3R)-pyrrolidin-3-ylcarbamoyl)-2-oxo-
oxazolidine-5-carboxamide
F
F
F ~ / ""N O ,CONH2
U ,,,
OMe N
~O
0O
The title compound was prepared by the procedure described in Example
67, followed by pressurized silica gel chromatography using an elution
system containing chloroform saturated with ammonia gas and
methanol.
F
F ,'"
F ~ / ""N ~ ,,,,C02Me
OMe N N
~O
//O
F
F
F ~ / ""N H2 . _
OMe H ~O
,/O
FAB MS: m/z=561 (M+H)
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Analysis: Calcd for C28H31N405F3
C 59.99, H 5.57, N 9.99.
Found: C 59.84, H 5.47, N 9.85.
The compounds of Examples 77- 79 were prepared using
procedures substantially as described above for Example 76.
EXAMPLE 77
(4S,5R)-tracns-4-(3,4-Difluorophenyl)-3-{1-[4-(4-fluorophenyl)cyclohexyl]-
(3R)-pyrrolidin-3-ylcarbamoyl]-2-oxo-oxazolidine-5-carboxamide
F
F
F ~ / .."N ~ ,CON H2
,,,
N N
~O
//O
FAB MS: m/z=531 (M+H)
Analysis: Calcd for C27H29N404F3~0.05H20
C 61.02, H 5.52, N 10.54.
Found: C 61.07, H 5.47, N 10.51.
EXAMPLE 78
(4S,5R)-trans-4-(3,4-Difluorophenyl)-3-{1-[4-(2-fluorophenyl)cyclohexyl]-
(3R)-pyrrolidin-3-ylcarbamoyl]-2-oxo-oxazolidine-5-carboxamide
F
F
/ .... N
F H _ -~ O
O
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Analysis: Calcd for C27H29N404F3
C61.12,H5.51,N10.56.
Found: C 61.20, H 5.67, N 10.46.
EXAMPLE 79
(4S,5R)-trans-3-{1-[4-(2-Cyano-4-fluorophenyl)cyclohexyl]-(3R)-pyrrolidin-
3-ylcarbamoyl)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-5-carboxamide
F
F
i
F ~ ~ ...,N ~ ,CONH2
~./ ,,,
N
CN H ~O
FAB MS: m/z=556 (M+H)
Analysis: Calcd for C28H28N5O4F3 ~ HCl ~ 0.85H20 ~ 0.35EtOAc
C 55.33, H 5.29, N 10.98.
Found: C 55.33, H 4.90, N 10.65.
EXAMPLE 80
Mixture of 4S-4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine and 4S-4-(3,4-difluorophenyl)-6-methoxymethyl-2-
oxo-2,3,4,5-tetrahydropyrimidine
H
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To a solution of (+)-4-(3,4-difluorophenyl)-6-methoxymethyl-
2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester (4.63 g,
14.7 mmol) in a methanol (100 ml) was added sodium hydroxide (2.94 g,
73.6 mmol). The resulting mixture was refluxed at 90 oC for 16 hours.
After cooling to room temperature the solvent was removed in vaccuo.
The solid was dissolved in CH2C12 and H20 then neutralized with 10%
aqueous HCl solution. The organic layer was dried over Na2S04,
concentrated, and purified by PCTLC (7% MeOH in CHC13 with 2%
NH40H) to afford a 2.65 g mixture of the title compounds (71°lo
yield).
The 'H NMR was consistent with the assigned structure.
MS (FAB) 255 (M+1)
EXAMPLE 81
Mixture of 4S-4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine and 4S-4-(3,4-difluorophenyl)-6-methoxymethyl-2-
oxo-2,3,4,5-tetrahydropyrimidine
O
H
To a solution of (+)-4-(3,4-difluorophenyl)-6-methoxymethyl-
2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester (5.36 g,
17.0 mmol) in a methanol (150 ml) was added 1N NaOH (10 ml). The
resulting mixture was refluxed at 90 oC for 16 hours. After cooling to
room temperature the solvent was removed in vacuo. The solid was
dissolved in CHZCl2 and H20 then neutralized with 10°lo aqueous HCl
solution. The organic layer was dried over Na2S04, concentrated, and
purified by PCTLC (7% MeOH in CHC13 with 2% NH40H) to afford a 2.35
g mixture of the title compounds (54% yield). The 'H NMR was
consistent with the assigned structure.
MS (FAB) 255 (M+1)
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EXAMPLE 82
4S-4-(3,4-Difluorophenyl)-6-methoxymethyl-3-(4-nitrophenoxycarbonyl)-2-
oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester
OZN ~ \ C
O
H
The title compound was prepared by treating the mixture
obtained from Example 80 or Example 81 (1.93 g, 7.59 mmol) with
lithium diisopropylamide (2.0M THF solution, 1.1 equivalents) in THF at
-78 °C for 20 minutes followed by the rapid addition of 4-nitrophenyl
chloroformate (1.5 equivalents) in THF. 0.488 g of the title compound
was obtained in a 15°~o yield. The 'H NMR was consistent with the
assigned structure.
EXAMPLE 83
Mixture of 4R-4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine and 4R-4-(3,4-difluorophenyl)-6-methoxymethyl-2-
oxo-2,3,4,5-tetrahydropyrimidine
F F
\ F \ F
0
O N ~ O N
H
The title compounds were prepared from 4R-4-(3,4-
difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-
carboxylic acid methyl ester (5.0 g, 17.7 mmol) using the procedure
described in Example 80. A mixture of 2.0 g of the title compounds was
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obtained in 50% yield. The 'H NMR was consistent with the assigned
structure.
MS (FAB) 255 (M+1)
Compounds of the invention can be prepared by reacting the
products obtained in Example 82 with an aminopiperidine, e.g., in
accordance with Scheme 3. Compounds of the invention can also be
prepared by preparing a nitrophenoxy derivative of the compound of
Example 83 in accordance with the procedure set forth in Example 82
and then reacting the derivative with an aminopiperidine in accordance
with Scheme 3.
The following compounds were prepared in accordance with
procedures set forth in the preceding Examples and Schemes.
EXAMPLE 84
trc~ns-(4S)-3-{ 1-(4-(2-Methoxyphenyl )cyclohexyl] azetidin-3-
ylmethylcarbamoyl)-4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester
hydrochloride
O
/ ,,,,, / O /
O
N~~N " ,NH
~''~~~0
O
1H NMR (CDC13, 400 MHz) consistent with assigned
structure. - --
FABLRMS m/e 613.4 g/mole (M++H, C32H38F2N406 =
612.67 g/mole. )
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HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04) - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 97°lo pure.
Anal. Calcd for C32H38F2N406 ~ 1.0 HCl, 0.3 CHC13 and
0.35 H20: C = 57.45, H = 5.97, N = 8.30. Found: C = 57.48, H = 5.97, N =
8.40.
EXAMPLE 85
trans-(4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid(1-[4-
(2-methoxyphenyl)-cyclohexyl]azetidin-3-ylmethyl-amide hydrochloride
F F
O~ _
/ -.,,,
N H O
'~ N
O
O
1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
FABLRMS m/e 500.3 g/mole (M++H, C27H31F2N304 =
499.45 g/mole. )
HPLC (Vydac; C 18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 99% pure.
Anal. Calcd for C27H31F2N304 ~ 1.35 HCl and 1.1 Et20: C =
59.83, H = 6.93, N = 9.56. Found: C = 60.17, H = 6.57, N = 6.28.
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EXAMPLE 86
traps-(4S)-3-{1-[4-(2-Cyanophenyl)cyclohexyl)azetidin-3-
ylmethylcarbamoyl}-4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester
hydrochloride
structure.
g/mole.)
CN F O
/ .,,, / O /
O
N~~N " ,NH
~'~O
O
1H NMR (CDCl3, 400 MHz) consistent with assigned
FABLRMS m/e 608 g/mole (M++H, C32H35F2N505 = 607.65
HPLC (Vydac; C 18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 97°l° pure.
Anal. Calcd for C32H35F2N505 ' 1.0 HCl and 0.8 H20: C =
58.36, H = 5.76, N = 10.64. Found: C = 58.40, H = 5.80, N = 10.42.
EXAMPLE 87
traps-(4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid {1-
[4-(2-cyanophenyl)-cyclohexyl]azetidin-3-ylmethyl-amide hydrochloride
CN _
/ ~.,,,
N H O . __
~~N
O
O
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1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
g/mole. )
FABLRMS m/e 495 g/mole (M++H, C27H28F2N403 = 494.54
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CHgCN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; >97% pure.
Anal. Calcd for C27H28F2N403 ~ 1.0 HCl and 0.85 H20: C =
59.36, H = 5.66, N = 10.26. Found: C = 59.48, H = 5.84, N = 9.87.
EXAMPLE 88
(High Rfj (4S)-3-{1-[4-(2-pyridyl)cyclohexyl]azetidin-3-lmethylcarbamoyl}-
4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester hydrochloride
F O O
N ~ ~ O
'
N'~ H ,,"N H
N
O O
structure.
g/mole.)
1H NMR (CDC13, 400 MHz) consistent with assigned
FABLRMS m/e 584 g/mole (M++H, C3pH35F2N5O5 = 583.63
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 99% pure.
Anal. Calcd for C3pH35F2N5O5 ~ 2.0 HCl, 0.5 CHC13 and
0.75 Et20: C = 52.19, H = 5.75, N = 9.09. Found: C = 52.21, H = 5.88, N '_
9.12.
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EXAMPLE 90
(High Rf) (4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid
{1-[4-(2-pyridyl)-cyclohexyl]azetidin-3-ylmethyl-amide hydrochloride
F F
N
N~.~ N ~O
O
O
1H NMR (CDCl3, 400 MHz) consistent with assigned
structure .
g/mol a . )
FABLRMS m/e 471 g/mole (M++H, C25H28F2N403 = 470.51
HPLC (Vydac; C 18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5°l° - 95%, over
16
minutes, 2 ml/min flow rate) focus = 215 nm; 99% pure.
Anal. Calcd for C25H28F2N403 ~ 2.0 HCI, 0.4 CHCl3 and
0.45 Et20: C = 52.31, H = 5.63, N = 8.97. Found: C = 52.33, H = 5.60, N =
9.15.
EXAMPLE 91
(Low Rf) (4S)-3-(1-[4-(2-pyridyl)cyclohexyl]azetidin-3-ylmethylcarbamoyl}-
4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester hydrochloride
F O O
ni ~ \ o
' ''~ . _
N'~ H ,,"N H
N
O O
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1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
g/mole. )
FABLRMS m/e 584 g/mole (M++H, C3pH35F2N505 = 583.63
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04) - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 99% pure.
Anal. Calcd for C3pH35F2N505 ~ 2.0 HCl and 2.35 Et20: C
= 51.63, H = 5.88, N = 10.04. Found: C = 51.63, H = 5.83, N = 10.20.
EXAMPLE 92
(Low Rf) (4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid
I1-[4-(2-pyridyl)-cyclohexyl]azetidin-3-ylmethyl-amide hydrochloride
F F
N
N H O
'~ N
O
structure.
g/mole. )
1H NMR (CDC13, 400 MHz) consistent with assigned
FABLRMS m/e 471 g/mole (M++H, C25H28F2N403 = 470.51
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95°l0 - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 99% pure.
Anal. Calcd for C25H28F2N403 ~ 2.0 HCI, 0.15 CHCl3 and
1.5 H20: C = 51.34, H = 5.68, N = 9.52. Found: C = 51.34, H = 5.63, N =
9.64.
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EXAMPLE 93
trans-(4S)-3-(1-[4-(2-Cyanophenyl)cyclohexyl]azetidin-3-ylcarbamoyl}-4-
(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester
\ CN
/ ~.,
N
"NH
O
~ H
1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
FABLRMS m/e 594 g/mole (M++H, C31H33F2N505 = 593.63
g/mole.)
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5°l0 - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 100% pure.
Anal. Calcd for C31H33F2N505 ~ 0.3 H20: C = 62.15, H =
5.65, N = 11.69. Found: C = 62.19, H = 5.75, N = 11.39.
EXAMPLE 94
trans-(4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid {1-
[4-(2-cyanophenyl)-cyclohexyl]azetidin-3-yl-amide
CN
\, F
~ F
/ __
N
'NH
O
O
O
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1H NMR (CDCl3, 400 MHz) consistent with assigned
structure.
g/mole. )
FABLRMS m/e 481 g/mole (M++H, C26H26F2N403 = 480.51
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1°~o H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 100% pure.
Anal. Calcd for C26H26F2N403 ' 0.2 CHC13 and 0.2 H20: C
= 63.84, H = 5.44, N = 11.43. Found: C = 63.85, H = 5.30, N = 11.41.
ZO
EXAMPLE 95
tracns-(4S)-3-( 1-[4-(2-Methoxyphenyl)cyclohexyl]azetidin-3-ylcarbamoyl ~-4-
(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester
\ O-
,,,,~ F
F
N
O
N I O~
O /~ N .,
O H
structure.
g/mole.)
1H NMR (CDC13, 400 MHz) consistent with assigned
FABLRMS m/e 599 g/mole (M++H, C31H36F2N406 = 598.64
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5°l0 - 95%, over 16
minutes, 2 mUmin flow rate) focus = 215 nm; 100% pure. - -
Anal. Calcd for C31H36F2N406~ C = 62.19, H = 6.06, N =
9.35. Found: C = 62.18, H = 5.96, N = 9.28.
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EXAMPLE 96
trans-(4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid {1-
[4-(2-methoxyphenyl)-cyclohexyl]azetidin-3-yl-amide
NH
O
O
O
1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
FABLRMS m/e 486 g/mole (M++H, C26H2gF2N304 = 485.53
g/mol e. )
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 100% pure.
Anal. Calcd for C26H2gF2N304: C = 64.31, H = 6.02, N =
8.65. Found: C = 64.28, H = 6.04, N = 8.68.
EXAMPLE 97
(Low Rf) (4S)-3-{1-[4-(2-pyridyl)cyclohexyl]azetidin-3-ylcarbamoyl)-4-(3,4-
difluorophenyl)-6-methoxymethyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-
carboxylic acid methyl ester hydrochloride
( ~ F
i F
N I \ . _
~ O
" 'NH i
~N I ~O
O O~ N
H
O-
I ~ ,,,, F
. F
I
N
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1H NMR (CDCl3, 400 MHz) consistent with assigned
structure.
g/mole. )
FABLRMS m/e 570 g/mole (M++H, C2gH33F2N505 = 466.577
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1°lo H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 100% pure.
Anal. Calcd for C29H33F2N505: C = 61.12, H = 6.04, N =
11.88. Found: C = 61.14, H = 5.90, N = 11.88.
EXAMPLE 98
(Lower Rf) (4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid
( 1-[4-(2-pyridyl)-cyclohexyl]azetidin-3-ylmethyl-amide
F
I
i F
N ~~ I /
'NH
~.- N l
o ~o
structure.
g/mole.)
1H NMR (CDC13, 400 MHz) consistent with assigned
FABLRMS m/e 457 g/mole (M++H, C24H26F2N404 = 456.49
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CHgCN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 100% pure.
Anai. Calcd for C24H26F2N404 ' 0.2 CHC13 and 0.45
EtOAc: C = 60.05, H = 5.78, N = 10.77. Found: C = 59.92, H = 5.77, N =
10.79.
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EXAMPLE 99
trans-(4S)-3-{1-[4-(2-Carboxymethoxyphenyl)cyclohexyl]azetidin-3-
ylmethylcarbamoyl}-4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester
structure.
g/mole. )
\ F
/ .,,,, F
\
N H / O
~N I O
O ~ N O
O H
1H NMR (CDC13, 400 MHz) consistent with assigned
FABLRMS m/e 641 g/mole (M++H, C33H38F2N4O7 = 640.69
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1°~o H3P04] - CH3CN, 95°l0 - 5°~0, 5% -
95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 97°~o pure.
Anal. Calcd for C33H38F2N4O7 ~ 0.2 CHCl3 and 0.25
EtOAc: C=59.82,H=5.90,N=8.16. Found:C=59.81,H=5.80,N=8.13.
EXAMPLE 100
trans-(4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid {1-
[4-(2-carboxymethoxyphenyl)-cyclohexyl]azetidin-3-ylmethyl-amide
F
/ ,,,,, F
O ( /
N N . _.
N
O ~O
0O
-168 -
EtOAc: C = 60.05, H =
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1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
g/mole. )
FABLRMS m/e 528 g/mole (M++H, C28H31F2N3O5 = 527.57
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1°~o H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 98% pure.
Anal. Calcd for C28H31F2N305 ~ 0.35 CHC13 and 0.1
EtOAc: C = 59.72, H = 5.61, N = 7.27. Found: C = 59.72, H = 5.78, N = 7.37.
E~~AMPLE 101
trans-(4S)-3-{1-[4-(2-Carboxyethoxyphenyl)-cyclohexyl]azetidin-3-
ylmethylcarbamoyl)-4-(3,4-difluorophenyl)-6-methoxymethyl-2-oxo-
1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester
\ F
/ ,,,, F
O N H / O
i
~N ~ o
O ~N O
O H
structure.
g/mole.)
1H NMR (CDC13, 400 MHz) consistent with assigned
FABLRMS m/e 655 g/mole (M++H, C34H40F2N407 = 654.71
HPLC (Vydac; C 18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95°~0, over 16
minutes, 2 mUmin flow rate) focus = 215 nm; 98% pure.
Anal. Calcd for C34H40F2N407 ~ 0.3 EtOAc: C = 62.06, H
6.27, N = 8.23. Found: C = 61.91, H = 6.09, N = 8.25.
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EXAMPLE 102
traps-(4S)-3-(1-[4-(2-Carboxyethoxyphenyl)-cyclohexyl]azetidin-3-
ylmethylcarbamoyl}-4-(3,4-difluorophenyl)-2-oxo-1,2,3,4-tetrahydro-4H-
furo[3,4-d]pyrimidine
F
/ .,,,, F
O N H / O
~N ( O
O ~N
O H
1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
FABLRMS m/e 609 g/mole (M++H, C32H34F2N406 = 608.64
g/mole. )
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1°~o H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 98% pure.
Anal. Calcd for C32H34F2N4O6 ' 0.4 EtOAc: C = 62.67, H =
5.82, N = 8.70. Found: C = 62.31, H = 5.71, N = 8.83.
EXAMPLE 103
traps-(4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic acid (1-
[4-(2-carboxyethoxyphenyl)-cyclohexyl]azetidin-3-ylmethyl-amide
F
/ .,, F
O I /
O N~ N
~N
O ~O
0O
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1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
g/mole. )
FABLRMS m/e 542 g/mole (M++H, C2gH33F2N305 = 541.59
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CHgCN, 95% - 5°~0, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 98% pure.
Anal. Calcd for C2gH33F2N305 ~ 0.3 EtOAc: C = 63.85, H =
6.28, N = 7.40. Found: C = 63.54, H = 6.12, N = 7.43.
EXAMPLE 104
(Diast A) (4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic
acid(1-[4-(2-pyridyl)cyclohexyl]-3-hydroxy-pyrrolidin-4-yl}amide
N ''
~N .,,,OH
~O
structure.
1H NMR (CDC13, 400 MHz) consistent with assigned
FABLRMS m/e 487.2 g/mole (M++H, C25H28F2N404 =
486.51 g/mole.)
HPLC (Vydac; C18; diameter = 4.6 mm; length = I50 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 100% pure.
EXAMPLE 105
(Diast B) (4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic
acid( 1-[4-(2-pyridyl)cyclohexyl]-3-hydroxy-pyrrolidin-4-yl}amide
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N ~~ F
F
N ..,,OH I
H
N
O ~O
,IO
structure.
1H NMR (CDC13, 400 MHz) consistent with assigned
FABLRMS m/e 487.2 glmole (M++H, C25H28F2N4O4 =
486.51 g/mole.)
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 100% pure.
EXAMPLE 106
(Diast A) (4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic
acid( 1-[4-(2-cyanophenyl)cyclohexyl]-3-hydroxy-pyrrolidin-4-yl}amide
I ~ cN
F
F
N ..,,OH
H
,/ N
p ~O
lIO
1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
g/mole. )
FABLRMS m/e 511 g/mole (M++H, C27H28F2N404 = 510.54
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HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 99% pure.
Anal. Calcd for C27H28F2N404 ~ 0.5 H20 and 0.25 Et20: C
= 62.50, H = 5.90, N = 10.41. Found: C = 62.55, H = 5.69, N = 10.36.
EXAMPLE 107
(Diast B) (4S)-4-(3,4-Difluorophenyl)-2-oxo-oxazolidine-3-carboxylic
acidf 1-[4-(2-cyanophenyl)cyclohexyl]-3-hydroxy-pyrrolidin-4-yl}amide
I w cN
,,
F
O
1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
FABLRMS m/e 511 g/mole (M++H, C27H28F2N404 = 510.54
g/mole. )
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 mI/min flow rate) focus = 215 nm; 99% pure.
Anal. Calcd for C27H28F2N404 ~ 0.4 H20 and 0.4 Et20: C =
62.75, H = 6.04, N = 10.24. Found: C = 62.77, H = 5.71, N = 10.23.
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EXAMPLE 108
(Racemic C~ pyrrolidine) (4S)-3-{1-[4-(2-Pyridyl)-cyclohexyl]-3-hydroxy-
pyrrolidin-4-ylcarbamoyl)-4-(3,4-difluorophenyl)-6-methyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester
I %~... F
N ,~ F
N ..".OH I /
O
H // N ( Oi
O
O N
H
structure.
g/mole. )
1H NMR (CDCl3, 400 MHz) consistent with assigned
FABLRMS m/e 600 g/mole (M++H, C3pH35F2N5O6 = 599.63
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95°l0 - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 98% pure.
Anal. Calcd for C3pH35F2N506 ~ 0.55 H20 and 0.35 Et20: C
= 59.34, H = 6.28, N = 11.02. Found: C = 59.32, H = 5.99, N = 10.98.
EXAMPLE 109
tracns-(4S,5S)-4-(3,4-Difluorophenyl)-5-methyl-2-oxo-oxazolidine-3-
carboxylic acid {1-[4-(2-cyanophenyl)-cyclohexyl]3-hydroxy-azetidin-3-
ylmethyl-amide
CN
F
/ ~~..
F __
I/
N H
N ,,~
OH N
O ~O
lIO
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1H NMR (CDCI3, 400 MHz) consistent with assigned
structure.
FABLRMS m/e 525.0 g/mole (M++H, C28H3pF2N404 =
524.56 g/mole.)
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 99% pure.
Anal. Calcd for C28H3pF2N404 ~ 0.40 H20 and 1.30
CH2Cl2: C = 54.80, H = 5.24, N = 8.73. Found: C = 54.78, H = 5.25, N =
8.67.
EXAMPLE 110
trans-(4S,5S)-4-(3,4-Difluorophenyl)-5-methyl-2-oxo-oxazolidine-3-
carboxylic acid (1-[4-(2-pyridyl)-cyclohexyl]3-hydroxy-azetidin-3-ylmethyl-
amide
F
F
N~ H ,,,.
~N
O
O
O
1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
FABLRMS m/e 485 g/mole (M++H, C26H30F2N403 = 484.54
g/mole. )
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95°l0 - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 97% pure. _ __
Anal. Calcd for C26H3pF2N403 ~ 0.15 CH2C12: C = 63.15, H
= 6.14, N = 11.27. Found: C = 63.40, H = 6.00, N = 11.24.
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EXAMPLE 111
trnns-(4S,5S)-4-(3,4-Difluorophenyl)-5-methyl-2-oxo-oxazolidine-3-
carboxylic acid {1-[4-(2-cyanophenyl)-cyclohexyl]azetidin-3-yimethyl-
amide
CN
I F F'
/ i...
I/
_ N~ H
N
~N
O// /I O
O
1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
FABLRMS m/e 509 g/mole (M++H, C28H3pF2N403 = 508.56
g/mole. )
HPLC (Vydac; C18; diameter = 4.6 mm; length = I50 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95°!0 - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 97% pure.
Anal. Calcd for C28H3pF2N403 ~ 0.05 Et20 and 0.10
CH2C12: C = 65.27, H = 5.94, N = 10.76. Found: C = 65.36, H = 65.36, N =
10.73.
EXAMPLE 112
trc~ns-(4~)-3-{ 1-[4-(2-Cyanophenyl)cyclohexyl]azetidin-3-
ylmethylcarbamoyl}-4-(3,4-difluorophenyl)-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester
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~N I O~
O O~ N
H
structure.
g/mole.)
CN
.,,, ~ F
N H O
N
1H NMR (CDC13, 400 MHz) consistent with assigned
FABLRMS m/e 564 g/mole (M++H, C30H31F2N504 = 563.60
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 98% pure.
Anal. Calcd for C3pH31F2N5O4 ' 0.15 Et20 and 0.15
CH2C12: C = 62.86, H = 5.63, N = 11.92. Found: C = 62.88, H = 5.74, N =
11.99.
EXAMPLE 113
trans-(4~)-3-{1-[4-(2-Cyanophenyl)cyclohexyl]3-hydroxy-azetidin-3-
ylmethylcarbamoyl}-4-(3,4-difluorophenyl)-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester
CN
N H
N il Oi
OH p
n
structure.
1H NMR (CDC13, 400 MHz) consistent with assigned
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FABLRMS m/e 580 g/mole (M++H, C3pH31F2N5O5 = 579.60
g/mole. )
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5°~0, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 98% pure.
Anal. Calcd for C3pH31F2N5O5 ' 0.05 CH2C12: C = 61.81, H
= 5.37, N = 12.00. Found: C = 61.61, H = 5.57, N = 12.22.
EXAMPLE 114
tracns-(4~)-3-{ 1-[4-(2-Pyridyl)cyclohexyl]3-hydroxy-azetidin-3-
ylmethylcarbamoyl)-4-(3,4-difluorophenyl)-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carboxylic acid methyl ester
F
1 ~~,,, w F
N I/
N O
~N N O'
O N
O H
1H NMR (CDC13, 400 MHz) consistent with assigned
structure.
g/mole.)
FABLRMS m/e 540 g/mole (M++H, C28H31F2N504 = 539.58
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm;
gradient = H20 [0.1% H3P04] - CH3CN, 95% - 5%, 5% - 95%, over 16
minutes, 2 ml/min flow rate) focus = 215 nm; 95% pure.
Anal. Calcd for C28H31F2N504 ~ 0.05 Et20 and 0.45
CH2C12: C = 59.17, H = 5.62, N = 12.04. Found: C = 59.18, H = 5.62, N =
12.04. - _
EXAMPLE I15
trans-(3R,4S,5S)-4-(3,4-Difluorophenyl)-5-methyl-2-oxo-oxazolidine-3-
carboxylic acid {1-[4-(2-pyridyl)-cyclohexyl]azetidin-3-ylmethyl-amide
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F
.,..C
structure.
g/mole. )
1H NMR. (CDCl3, 400 MHz) consistent with assigned
FABLRMS m/e 485 g/mole (M++H, C2gHgOFgN4O3 = 484.548
HPLC (Vydac; C18; diameter = 4.6 mm; length = 150 mm; gradient
= H20 [0.1°Jo H3POqJ - CH3CN, 95°.b - 5%, b% - 95%, over I6
minutes, 2
ml/min flow rate) focus = 215 nm; 96.4% pure.
EXAMPLE 116
As a specific embodiment of an oral composition, 100 mg of
the compound of Example 1 is formulated with suffident finely divided
lactose to provide a total amount of 580 to b90 mg to fill a size O hard gel
capsule.
EXAMPLE 117
Screening aseaw A~nha la Adre-eyS..~!~~~t~r 3ind;ne
Membranes prepared from the etably transfected human
alpha la cell Iine (ATCC CRL 11140) were used to identify compounds
that bind to the human alpha la adrenergic receptor. These competition
binding reactions (total volume = 200 ~1) contained 50 mM Tris-HCl pH.
7.4, 5 mM EDTA, 150 mM NaCI, 100 pM [I25 I]-HEAT, membraaes
prepared from the sipha la cell line and increasing amounts of _
unlabeled ligand. Reactions were incubated at room temperature for
one hour with shalang. Reactions were filtered onto Whatman GF/C
glass fiber filters arith a Inotec 96 well cell harvester. Filters were
washed three times with ice cold buffer and bound radioactivity was
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determined (Ki). Representative compounds of the present invention
were found to have Ki values _< 50 nM.
EXAMPLE 118
Selective Bindine assays
Membranes prepared from stably transfected human alpha
1d and alpha lb cell lines (ATCC CRL 11138 and CRL 11139, respectively)
were used to identify compounds that selectively bind to the human
alpha la adrenergic receptor. These competition binding reactions (total
volume = 200 ~1) contained 50 mM Tris-HCl pH. 7.4, 5 mM EDTA, 150
mM NaCI, 100 pM [125 I~_gEAT, membranes prepared from cell lines
transfected with the respective alpha 1 subtype expression plasmid and
increasing amounts of unlabeled ligand. Reactions were incubated at
room temperature for one hour with shaking. Reactions were filtered
onto Whatman GF/C glass fiber filters with a Inotec 96 well cell
harvester. Filters were washed three times with ice cold buffer and
bound radioactivity was determined (Ki).
All of the compounds of the present invention prepared in
the foregoing Examples were found to have alpha la Ki values of less
than 50 nM as determined via the screening assay described in Example
117, except for Examples 12 (<200 nM), 16 (ca. 800 nM) and 20 (inactive).
All of the compounds were further found to be at least about 50-fold more
selective in binding to alpha la receptors versus binding to the alpha lb
and alpha ld receptors, as determined via the selective binding assay
described in the preceding paragraph, except for Examples 12 (at least
about 3-fold), 16 (at least about 8-fold), 20, and 78 (about 37-fold). All of
the
compounds except for those of Examples 8, 12, 14, 16, 19, 20, 21, 24, 30, 40,
47, 50, 51, 52, 67, 69, 70, 76, 77 and 78 were at least about 100-fold more
selective in binding to alpha la receptors versus alpha lb and ld
receptors.
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EXAMPLE 119
EXEMPLARY COUNTERSCREENS
1. Assay Title: Dopamine D2, D3, D4 in vitro screen
Obiective of the Assay:
The objective of this assay is to eliminate agents which
specifically affect binding of [3H] spiperone to cells expressing human
dopamine receptors D2, D3 or D4.
Method:
Modified from VanTol et al (1991); Nature (Vol 350) Pg 610-
613.
Frozen pellets containing specific dopamine receptor
subtypes stably expressed in clonal cell lines are lysed in 2 ml lysing
buffer (lOmM Tris-HCl/5mM Mg, pH 7.4). Pellets obtained after
centrifuging these membranes (15' at 24,450 rpm) are resuspended in
50mM Tris-HCl pH 7.4 containing EDTA, MgCI[2], KCl, NaCI, CaCl[2]
and ascorbate to give a 1 Mg/mL suspension. The assay is initiated by
adding 50-75 ~g membranes in a total volume of 500 ~.1 containing 0.2 nM
[3H]-spiperone. Non-specific binding is defined using 10 ~.M
apomorphine. The assay is terminated after a 2 hour incubation at room
temperature by rapid filtration over GFB filters presoaked in 0.3% PEI,
using 50mM Tris-HCl pH 7.4.
2. Assay Title: Serotonin 5HTla
Obiective of the Assay
The objective of this assay is to eliminate agents which
specifically affect binding to cloned human 5HTla receptor
Met o
Modified from Schelegel and Peroutka Biochemicacl
Pha~rmaccology 35: 1943-1949 (1986).
Mammalian cells expressing cloned human 5HTla
receptors are lysed in ice-cold 5 mM Tris-HCl , 2 mM EDTA (pH 7.4) and
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homogenized with a polytron homogenizer. The homogenate is
centrifuged at 1000Xg for 30', and then the supernatant is centrifuged
again at 38,OOOXg for 30'. The binding assay contains 0.25 nM [3H]8-OH-
DPAT (8-hydroxy-2-dipropylamino-1,2,3,4-tetrahydronaphthalene) in 50
mM Tris-HCl, 4 mM CaCl2 and lmg/ml ascorbate. Non-specific binding
is defined using 10 ~.M propranolol. The assay is terminated after a 1
hour incubation at room temperature by rapid filtration over GF/Cfilters
EXAMPLE 120
EXEMPLARY FUNCTIONAL ASSAYS
In order to confirm the specificity of compounds for the
human alpha la adrenergic receptor and to define the biological activity
of the compounds, the following functional tests may be performed:
1. In vitro Rat, Dog and Human Prostate and Dog Urethra
Taconic Farms Sprague-Dawley male rats, weighing 250-
400 grams are sacrificed by cervical dislocation under anesthesia
(methohexital; 50 mg/kg, i.p.). An incision is made into the lower
abdomen to remove the ventral lobes of the prostate. Each prostate
removed from a mongrel dog is cut into 6-8 pieces longitudinally along
the urethra opening and stored in ice-cold oxygenated Krebs solution
overnight before use if necessary. Dog urethra proximal to prostate is
cut into approximately 5 mm rings, the rings are then cut open for
contractile measurement of circular muscles. Human prostate chips
from transurethral surgery of benign prostate hyperplasia are also
stored overnight in ice-cold Krebs solution if needed.
The tissue is placed in a Petri dish containing oxygenated
Krebs solution [NaCl, 118 mM; KCl, 4.7 mM; CaCl2, 2.5 mM; KH2P04,
1.2 mM; MgS04, 1.2 mM; NaHC03, 2.0 mM; dextrose, 11 mM] warmed
to 37°C. Excess lipid material and connective tissue are carefully . _
removed. Tissue segments are attached to glass tissue holders with 4-0
surgical silk and placed in a 5 ml jacketed tissue bath containing Krebs
buffer at 37°C, bubbled with 5% C02/95% 02. The tissues are connected
to a Statham-Gould force transducer; 1 gram (rat, human) or 1.5 gram
(dog) of tension is applied and the tissues are allowed to equilibrate for
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one hour. Contractions are recorded on a Hewlett-Packard 7700 series
strip chart recorder.
After a single priming dose of 3 p.M (for rat), 10 ~.M (for dog)
and 20 ~.M (for human) of phenylephrine, a cumulative concentration
response curve to an agonist is generated; the tissues are washed every
minutes for one hour. Vehicle or antagonist is added to the bath and
allowed to incubate for one hour, then another cumulative concentration
response curve to the agonist is generated.
ECSp values are calculated for each group using GraphPad
10 Inplot software. pA2 (-log Kb) values were obtained from Schild plot
when three or more concentrations were tested. When less than three
concentrations of antagonist are tested, Kb values are calculated
according
to the following formula Kb =~jB ,
x-1
where x is the ratio of ECSp of agonist in the presence and absence of
antagonist and [B] is the antagonist concentration.
2. Measurement of Intra-Urethral Pressure in Anesthetized Dogs
PURPOSE: Benign prostatic hyperplasia causes a decreased urine flow
rate that may be produced by both passive physical obstruction of the
prostatic urethra from increased prostate mass as well as active
obstruction due to prostatic contraction. Alpha adrenergic receptor
antagonists such as prazosin and terazosin prevent active prostatic
contraction, thus improve urine flow rate and provide symptomatic
relief in man. However, these are non-selective alpha 1 receptor
antagonists which also have pronounced vascular effects. Because we
have identified the alpha la receptor subtype as the predominent subtype
in the human prostate, it is now possible to specifically target this
receptor to inhibit prostatic contraction without concomitant changes-in
the vasculature. The following model is used to measure adrenergically
mediated changes in intra-urethral pressure and arterial pressure in
anesthetized dogs in order to evaluate the efficacy and potency of
selective alpha adrenergic receptor antagonists. The goals are to: 1)
identify the alpha 1 receptor subtypes responsible for prostatic/urethral
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contraction and vascular responses, and 2) use this model to evaluate
novel selective alpha adrenergic antagonists. Novel and standard alpha
adrenergic antagonists may be evaluated in this manner.
METHODS: Male mongrel dogs (7-12 kg) are used in this study. The
dogs are anesthetized with pentobarbital sodium (35 mg/kg, i.v. plus 4
mg/kg/hr iv infusion). An endotracheal tube is inserted and the animal
ventilated with room air using a Harvard instruments positive
displacement large animal ventilator. Catheters (PE 240 or 260) are
placed in the aorta via the femoral artery and vena cava via the femoral
veins (2 catheters, one in each vein) for the measurement of arterial
pressure and the administration of drugs, respectively. A supra-pubic
incision --1/2 inch lateral to the penis is made to expose the urethers,
bladder and urethra. The urethers are ligated and cannulated so that
urine flows freely into beakers. The dome of the bladder is retracted to
facilitate dissection of the proximal and distal urethra. Umbilical tape is
passed beneath the urethra at the bladder neck and another piece of
umbilical tape is placed under the distal urethra approximately 1-2 cm
distal to the prostate. The bladder is incised and a Millar micro-tip
pressure transducer is advanced into the urethra. The bladder incision
is sutured with 2-0 or 3-0 silk (purse-string suture) to hold the
transducer. The tip of the transducer is placed in the prostatic urethra
and the position of the Millar catheter is verified by gently squeezing the
prostate and noting the large change in urethral pressure.
Phenylephrine, an alpha 1 adrenergic agonist, is
administered (0.1-100 ug/kg, iv; 0.05 ml/kg volume) in order to construct
dose response curves for changes in intra-urethral and arterial
pressure. Following administration of increasing doses of an alpha
adrenergic antagonist (or vehicle), the effects of phenylephrine on
arterial pressure and intra-urethral pressure are re-evaluated. Four or
five phenylephrine dose-response curves are generated in each animal
(one control, three or four doses of antagonist or vehicle). The relative
antagonist potency on phenylephrine induced changes in arterial and
intra-urethral pressure are determined by Schild analysis. The family
of averaged c:.rves are fit simultaneously (using ALLFIT software
package) with a four paramenter logistic equation constraining the
slope, minimum response, and maximum response to be constant
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among curves. The dose ratios for the antagonist doses (rightward shift
in the dose-response curves from control) are calculated as the ratio of
the ED50's for the respective curves. These dose-ratios are then used to
construct a Schild plot and the Kb (expressed as ug/kg, iv) determined.
The Kb (dose of antagonist causing a 2-fold rightward shift of the
phenylephrine dose-response curve) is used to compare the relative
potency of the antagonists on inhibiting phenylephrine responses for
intra-urethral and arterial pressure. The relative selectivity is
calculated as the ratio of arterial pressure and intra-urethral pressure
Kb's. Effects of the alpha 1 antagonists on baseline arterial pressure are
also monitored. Comparison of the relative antagonist potency on
changes in arterial pressure and intra-urethral pressure provide
insight as to whether the alpha receptor subtype responsible for
increasing intra-urethral pressure is also present in the systemic
vasculature. According to this method, one is able to confirm the
selectivity of alpha la adrenergic receptor antagonists that prevent the
increase in intra-urethral pressure to phenylephrine without any
activity at the vasculature.
While the foregoing specification teaches the principles of
the present invention, with examples provided for the purpose of
illustration, it will be understood that the practice of the invention
encompasses all of the usual variations, adaptations and/or
modifications as come within the scope of the following claims and their
equivalents.
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