Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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PROCESS FOR THE SYNTHESIS OF (S)-3-AMINO-4-
(DIFLUOROMETHYLENYL)CYCLOPENT-1-ENE-1-CARBOXYLIC ACID
This invention was made with government support under RO1 DA030604 awarded by
the
National Institutes of Health. The government has certain rights in the
invention.
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims benefit and priority to U.S. Provisional Application
No. 62/676,373, filed
May 25, 2018; U.S. Provisional Application No. 62/814,026, filed March 5,2019;
and U.S. Provisional
Application No. 62/835,776, filed April 18, 2019, which are all incorporated
herein by reference in their
entireties.
TECHNICAL FIELD
Synthesis of (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic
acid.
BACKGROUND
(S)-3-Amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid is an
inhibitor of
y-aminobutyric acid aminotransferase (GABA-AT) and has been shown as a
possible treatment of
epilepsy, addiction and hepatocellular carcinoma. See, e.g., US Patent No.
9,670,141 and Juncosa et
al., I Am. Chem. Soc. 2018, 140, 2151-2164. GABA is an inhibitory
neurotransmitter in the central
nervous system (CNS). When GABA concentrations in the brain drop below a
threshold level,
convulsions can occur. Increasing GABA levels has been shown to stop
convulsions. Additionally,
increased concentrations of GABA antagonize the release of dopamine from the
nucleus accumbens, a
region of the hypothalamus associated with reward and motivation, and have
been suggested as a
possible treatment of addiction. Unfortunately, direct administration of GABA
is not viable as GABA
does not cross the blood brain barrier. GABA concentrations, however, can be
increased by inhibiting
GABA aminotransferase (GABA-AT). 4-Aminohex-5-enoic acid, also known as
vigabatrin (marketed
as Sabri10), currently is the only FDA approved inhibitor of GABA-AT for the
treatment of infantile
spasms and has been shown as a possible treatment of addiction. Vigabatrin,
however, requires a large
dose (1-3 g/day), inhibits multiple GABA receptors, and, with prolonged use,
causes retinal damage
in 25-40% of patients.
In vivo studies in rats indicate that (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic acid is superior to previous inhibitors of GABA-AT at suppressing
the release of dopamine
in the corpus striatum after exposure to cocaine or nicotine. (S)-3-Amino-4-
(difluoromethylenyl)cyclopent- 1-ene- 1-carboxylic acid does not inhibit off-
target aminotransferase
enzymes like alanine aminotransferase and aspartate aminotransferase. It
additionally does not inhibit
the hERG potassium ion channel or various microsomal cytochrome P450 enzymes.
1
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According existing techniques, (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-
ene-1-
carboxylic acid has been synthesized in six steps from (1S,3S)-3-amino-4-
(difluoromethylidene)cyclopentane-1-carboxylic acid (also known as CPP-115).
Na}04
(4 steps) seph Nal1CO3
F F
-$CO214 (57% yieico CO2H
C1/401''
CPP-115 Compound A
8 steps imm
COMMeitlei materiel
SH
HO2C
1110
F CO2H F CO2H F sAillb CO2
Ingt H
CIH3hr C1H3te C1H3N
Compound B
14 total steps
60% yd 36% yield 3.7% overall yield
(ohromatographicallY inseparable)
See also, e.g., Juncosa et al., I Am. Chem. Soc., supra, and US Patent Nos.
7,381,748, 6,794,413 and
9,670,141, each incorporated herein by reference in their entireties.
CPP-115 is an inhibitor of GABA-AT and currently in clinical trials for the
treatment of
epilepsy. It has been determined that (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic acid is 9.8 times more efficient as an activator of GABA-AT than
CPP-115. Since
synthesis of CPP-115 takes 8-steps, the total synthetic step count from
commercial starting material to
(S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid is
fourteen with an overall
yield of 3.7%. The synthesis of CPP-115 involves the use of the highly
flammable tert-butyl lithium
(on gram scale) to install the 1,1'-difluoroolefin, which limits the scale at
which the reaction can be
run. Furthermore, the existing synthesis relies of the introduction of the
cyclopentene through
selenoxide elimination. Protected CPP-115 is selenated in 70% yield, although
yields can vary
depending on scale. a-Elimination of Compound A yields a mixture of
chromatographically
inseparable isomers in a 5:3 ratio favoring (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic acid. Compound B is selectively degraded using thiosalicylic acid
to produce solely (S)-3-
amino-4-(difluoromethylenyl)cyclopent-1-ene- 1-carboxylic acid in an overall
yield of 36% from
Compound A. Only small batches of (S)-3-amino-4-(difluoromethylenyl)cyclopent-
l-ene-1-
carboxylic acid can be obtained using the existing technique. Additionally,
since selenium is toxic and
regulated by the FDA to levels below 80-150 jig/day, the production of selenol
in the penultimate step
complicates the synthesis and purification of (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic acid.
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Accordingly, there is a need for processes that are better suited for larger
scale preparation of
(S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid that will
reduce costs, decrease
the number of manufacturing steps, decrease hazardous environmental waste, and
increase efficiency
of manufacture.
SUMMARY
A process for preparing (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic
acid (1) or salt thereof is provided which includes converting ((1R,4S)-2-
Azabicyclo[2.2.11hept-5-en-
3-one) (2) to (1R,45)-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11hept-5-en-3-one
(3). (1R,45)-2-(4-
Methoxybenzy1)-2-azabicyclo[2.2.11hept-5-en-3-one (3) is converted to
(1R,4R,6S,7R)-7-Bromo-2-(4-
methoxybenzy1)-3-oxo-2-azabicyclo[2.2.11heptan-6-y1 acetate (4). (1R,4R,6S,7R)-
7-Bromo-2-(4-
methoxybenzy1)-3-oxo-2-azabicyclo[2.2.11heptan-6-y1 acetate (4) is converted
to (1R,4R,65,7R)-7-
Bromo-6-hydroxy-2-(4-methoxybenzy1)-2-azabicyclo[2.2.1.1heptan-3-one (25).
(1R,4R,65,7R)-7-
Bromo-6-hydroxy-2-(4-methoxybenzy1)-2-azabicyclo[2.2.1.1heptan-3-one (25) is
converted to
(1R,4R,7R)-7 -Bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione
(5). (1R,4R,7R)-7-
Bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione (5) is
converted to (1R,4R,7R)-7 -
Bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-azabicyclop.2.11heptan-3-one
(6). (1R,4R,7R)-
7-Bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (6) is converted
to (1R,4R,7R)-7-
bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7). (1R,4R,7R)-7-
Bromo-6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7) is converted to
(1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one
(8). (1R,4R,7R)-7 -
Bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-
3-one (8) is
converted to methyl (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-
carboxylate (9). Methyl (5)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-
carboxylate (9) is converted to (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-
ene-1-carboxylic acid
(1).
(S)-3-Amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1) is
made by a
process described herein. Compositions including (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-
ene-1-carboxylic acid (1) are described herein. Pharmaceutical compositions
including (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1) are described
herein.
(1R,45)-2-(4-Methoxybenzy1)-2-azabicyclo[2.2.11hept-5-en-3-one (3) is provided
herein.
Compositions including (1R,45)-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11hept-5-
en-3-one (3) are
provided herein. Pharmaceutical compositions including (1R,4S)-2-(4-
methoxybenzy1)-2-
azabicyclo[2.2.11hept-5-en-3-one (3) are provided herein. (1R,4S)-2-(4-
Methoxybenzy1)-2-
azabicyclo[2.2.11hept-5-en-3-one (3) is made by a process disclosed herein.
(1R,4R,6S,7R)-7 -Bromo-2-(4-methoxybenzy1)-3-oxo-2-azabicyclo[2.2.11heptan-6-
y1 acetate
(4) is provided herein. Compositions including (1R,4R,6S,7R)-7-bromo-2-(4-
methoxybenzy1)-3-oxo-
3
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2-azabicyc1o[2.2.11heptan-6-y1 acetate (4) are provided herein. Pharmaceutical
compositions
including (1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-
azabicyclo[2.2.11heptan-6-y1 acetate
(4) are provided herein. (1R,4R,6S,7R)-7-Bromo-2-(4-methoxybenzy1)-3-oxo-2-
azabicyclo[2.2.11heptan-6-y1 acetate (4) is made by a process disclosed
herein.
(1R,4R,6S,7R)-7-Bromo-6-hydroxy-2-(4-methoxybenzy1)-2-azabicyclo[2.2.1.1heptan-
3-one
(25) is provided herein. Compositions including (1R,4R,6S,7R)-7-Bromo-6-
hydroxy-2-(4-
me thoxybenzy1)-2-azabicyclo[2.2.1.1heptan-3-one (25) are provided herein.
Pharmaceutical
compositions including (1R,4R,6S,7R)-7-Bromo-6-hydroxy-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.1.1heptan-3-one (25) are provided herein. (1R,4R,6S,7R)-7-Bromo-
6-hydroxy-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.1.1heptan-3-one (25) is made by a process
disclosed herein.
(1R,4R,7R)-7 -Bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione
(5) is
provided herein. Compositions including (1R,4R,7R)-7-bromo-2-(4-methoxybenzy1)-
2-
azabicyclo[2.2.11heptane-3,6-dione (5) are provided herein. Pharmaceutical
compositions including
(1R,4R,7R)-7-bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione
(5) are provided
herein. (1R,4R,7R)-7 -Bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-
dione (5) is made
by a process described herein.
(1R,4R,7R)-7 -B romo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-
azabicyclop.2.11heptan-
3-one (6) is provided herein. Compositions including (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-
azabicyclo[2.2.11heptan-3-one (6) are provided herein. Pharmaceutical
compositions including
(1R, 4R,7 R)-7-bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one
(6)are provided herein.
(1R,4R,7R)-7 -Bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-
azabicyclop.2.11heptan-3-one
(6) is made by a process described herein.
(1R,4R,7R)-7-Bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7)
is provided
herein. Compositions including (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-
azabicyclo[2.2.11heptan-3-one (7) are provided herein. Pharmaceutical
compositions including
(1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7)
are provided herein.
(1R,4R,7R)-7 -Bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7)
is made by a process
described herein.
(1R,4R,7R)-7 -Bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-
azabicyclo[2.2.11heptan-3-one (8) is provided herein. Compositions including
(1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one
(8) are provided herein.
Pharmaceutical compositions including (1R,4R,7R)-7-bromo-6-(difluoromethylene)-
2-(tert-
butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one (8) are provided herein.
1R,4R,7R)-7-Bromo-6-
(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one
(8) is made by a
process described herein.
Methyl (S)-3-((tert-Butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-
1-
carboxylate (9) is provided herein. Compositions including methyl (S)-3-((tert-
4
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butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-carboxylate (9)
are provided herein.
Pharmaceutical compositions including methyl (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylate (9) are provided herein.
Methyl (S)-3-((tert-
Butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-carboxylate (9)
is made by a
process described herein.
(S)-3-((tert-Butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-
carboxylic acid
(19) is provided herein. Compositions including (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-l-ene-l-carboxylic acid (19) are provided herein.
Pharmaceutical
compositions including (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-
carboxylic acid (19) are provided herein. (S)-3-((tert-Butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-l-ene-l-carboxylic acid (19) is made by a process
described herein.
Compositions including (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic
acid (1) and (1S,3S)-3-amino-4-(difluoromethylidene)cyclopentane-l-carboxylic
acid are provided.
Pharmaceutical compositions including (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic acid (1) and (1S,3S)-3-amino-4-(difluoromethylidene)cyclopentane-l-
carboxylic acid are
provided.
Compositions including (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic
acid (1) and one or more of (1R,4S)-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11hept-5-en-3-one (3),
(1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-azabicyclo[2.2.11heptan-6-y1
acetate (4),
(1R,4R,6S,7R)-7-Bromo-6-hydroxy-2-(4-methoxybenzy1)-2-azabicyclo[2.2.1.1heptan-
3-one (25),
(1R,4R,7R)-7-bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione
(5), (1R, 4R,7R)-7-
bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (6), (1R,4R,7R)-7-
bromo-6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7), (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one
(8), methyl (S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-carboxylate (9),
or (S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid
(19) are provided.
A process for preparing (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic
acid (1) is provided which includes converting ethyl-cyclopent-3-ene-
carboxylate (10) to (3R,4S)-
ethy1-3-((tert-butoxycarbonyl)amino)-4-(hydroxy)cyclopentane carboxylate (11).
(3R,4S)-Ethy1-3-
((tert-butoxycarbonyl)amino)-4-(hydroxy)cyclopentane carboxylate (11) is
converted to (3R)-ethy1-3-
((tert-butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12). (3R)-Ethy1-3-
((tert-
butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate is converted to ethyl
(9S)-9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-spiro[4.4]nonanecarboxylate (13). Ethyl (9S)-
9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-spiro[4.4]nonanecarboxylate (13) is converted
to ethyl (S)-9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-spiro[4.4]nonene-7-carboxylate (14). Ethyl
(S)-9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-spiro[4.4]nonene-7-carboxylate (14) is
converted to ethyl (S)-3-
((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-
carboxylate (15). Ethyl (S)-3-
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((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-
carboxylate (15) is converted
to (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1).
(3R,4S)-Ethyl-3-((tert-butoxycarbonyl)amino)-4-(hydroxy)cyclopentane
carboxylate (11) is
provided herein. Compositions including (3R,45)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-
(hydroxy)cyclopentane carboxylate (11) are provided herein. Pharmaceutical
compositions including
(3R,4S)-ethyl-3-((tert-butoxycarbonyl)amino)-4-(hydroxy)cyclopentane
carboxylate (11) are provided
herein. (3R,4S)-Ethyl-3-((tert-butoxycarbonyl)amino)-4-(hydroxy)cyclopentane
carboxylate (11) is
made by a process described herein.
(3R)-Ethyl-3-((tert-butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12)
is provided
herein. Compositions including (3R)-ethyl-3-((tert-butoxycarbonyl)amino)-4-oxo-
cyclopentane
carboxylate (12) are provided herein. Pharmaceutical compositions including
(3R)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12) are provided herein.
(3R)-Ethy1-3-((tert-
butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12) is made by a process
described herein.
Ethyl (9S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-
spiro[4.41nonanecarboxylate (13) is
provided herein. Compositions including ethyl (9S)-9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-
spiro[4.41nonanecarboxylate (13) are provided herein. Pharmaceutical
compositions including ethyl
(9S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro[4.41nonanecarboxylate (13)
are provided herein.
Ethyl (9S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-
spiro[4.4]nonanecarboxylate (13) is made by a
process described herein.
Ethyl (S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro[4.41nonene-7-
carboxylate (14) is
provided herein. Compositions including ethyl (S)-9-(tert-butoxycarbonylamino)-
1,4-dioxa-7-
spiro[4.41nonene-7-carboxylate (14) are provided herein. Pharmaceutical
compositions including
ethyl (S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro[4.41nonene-7-
carboxylate (14) are provided
herein. Ethyl (S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro[4.41nonene-7-
carboxylate (14) is
made by a process described herein.
Ethyl (S)-3-((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-
1-
carboxylate (15) is provided herein. Compositions including ethyl (S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-l-ene-l-carboxylate (15)
are provided
herein. Pharmaceutical compositions including ethyl (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylate (15) are provided herein.
Ethyl (S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-l-ene-l-carboxylate (15)
is made by a
process described herein.
Compositions including (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic
acid (1) and one or more of ethyl-cyclopent-3-ene-carboxylate (10), (3R,45)-
ethy1-3-((tert-
butoxycarbonyl)amino)-4-(hydroxy)cyclopentane carboxylate (11), (3R)-ethy1-3-
((tert-
butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12), ethyl (9S)-9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-spiro[4.41nonanecarboxylate (13), ethyl (S)-9-
(tert-
6
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butoxycarbonylamino)-1,4-dioxa-7-spiro[4.41nonene-7-carboxylate (14), or ethyl
(S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-carboxylate (15),
are provided.
BRIEF DESCRIPTION OF DRAWINGS
FIG. 1 is a 500 MHz 1H NMR spectrum (CDC13) of (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-one (6).
FIG. 2 is a 13C NMR spectrum (CDC13; 126 MHz) of (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-one (6).
FIG. 3 is a 127.5 MHz19F NMR spectrum (CDC13) of (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-one (6).
FIG. 4 is a 500 MHz 1H NMR spectrum (CDC13) of (1R,4R,7R)-7-Bromo-6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7).
FIG. 5 is a 13C NMR spectrum (CDC13; 126 MHz) of (1R,4R,7R)-7-Bromo-6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7).
FIG. 6 is a 127.5 MHz19F NMR spectrum of (1R,4R,7R)-7-Bromo-6-
(difluoromethylene)-2-
azabicyclo[2.2.11heptan-3-one (7) (CDC13).
FIG. 7 is a 500 MHz III NMR spectrum (CDC13) of methyl (S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-l-ene-l-carboxylate (9).
FIG. 8 is a 13C NMR spectrum (CDC13; 126 MHz) of methyl (S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-l-ene-l-carboxylate (9).
FIG. 9 is a 127.5 MHz19F NMR spectrum of methyl (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-l-ene-l-carboxylate (9).
FIG. 10 is a 500 MHz 1H NMR spectrum (CDC13) (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1).
FIG. 11 is a 13C NMR spectrum (CDC13; 126 MHz) of (S)-3-amino-4-
(difluoromethylenyl)cyclopent-l-ene-l-carboxylic acid (1).
FIG. 12 is a 127.5 MHz19F NMR spectrum of (S)-3-amino-4-
(difluoromethylenyl)cyclopent-
l-ene-l-carboxylic acid (1).
DETAILED DESCRIPTION
Provided herein are processes, compounds and compositions for making (S)-3-
amino-4-
(difluoromethylenyl)cyclopent- 1-ene- 1-carboxylic acid (1). The processes
herein are scalable and
high yielding as compared to the existing technique, avoid the use of selenium
and tert-butyllithium,
and avoid forming multiple isomers from an a-elimination. In embodiments, the
present process
incorporates elimination of a leaving group from the 0-position, precludes a
resulting mixture of
7
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isomers, reduces the number of synthesis steps from fourteen to nine as
compared to the existing
technique, and increases yield from 3.7% to 8.1%.
In embodiments, synthesis of (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-
1-
carboxylic acid (1) is shown in Scheme 1 starting with ((1R,4S)-2-
azabicyclo[2.2.11hept-5-en-3-one)
(2).
Scheme 1
(31 Br Br
i 0 0 0
ts:17M
AcCL--1--CT __________________________
4 'PM
Pin 6 PMB
Br
0
\.õ -mak,
= __________________ air co,ti F
* CO2440 F - 14 N
CHO( POW' F F 7
9 8
AC=Acetyl, PMB= 4-Methoxybenzyl, PG= Protecting Group
According to Scheme 1, a process for preparing (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1) or salt thereof is
provided which includes
converting ((1R,45)-2-azabicyclo [2 .2.11hept-5-en-3-one) (2) to (1R,45)-2-(4-
methoxybenzy1)-2-
azabicyclo [2 .2.11hept-5-en-3-one (3). (1R,45)-2-(4-Methoxybenzy1)-2-
azabicyclo [2 .2.11hept-5-en-3-
one (3) is converted to (1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-
azabicyclo [2 .2.11heptan-6-y1 acetate (4). (1R,4R,6S,7R)-7-Bromo-2-(4-
methoxybenzy1)-3-oxo-2-
azabicyclo[2.2.11heptan-6-y1 acetate (4) is converted to (1R,4R,7R)-7-bromo-2-
(4-methoxybenzy1)-2-
azabicyclo [2 .2.11heptane-3,6-dione (5). (1R,4R,7R)-7 -Bromo-2-(4-
methoxybenzy1)-2-
azabicyclo [2 .2.11heptane-3,6-dione (5) is converted to (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-
(4-methoxybenzy1)-2-azabicyclo [2 .2.11heptan-3-one (6). (1R,4R,7R)-7-Bromo-6-
(difluoromethylene)
-2-(4-methoxybenzy1)-2-azabicyclo [2 .2.11heptan-3-one (6) is converted to
(1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-azabicyclo [2 .2.11heptan-3-one (7). (1R,4R,7R)-7-Bromo-
6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7) is converted to
(1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo [2 .2.11heptan-3-one
(8). (1R,4R,7R)-7 -
Bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo [2
.2.11heptan-3-one (8) is
converted to methyl (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-
carboxylate (9). Methyl (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-
carboxylate (9) is converted to (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-
ene-1-carboxylic acid
(1).
In embodiments, synthesis of (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-
1-
carboxylic acid (1) is shown in Scheme 1A.
8
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Scheme 1A
Br
a. MASON MI, NW 0 c. KzO03 WON a *.
2i)KOliAt 0
b. MOW It. TPAP NMO al A HO
N 144
'Fivis 02 %) 'pms %) c's-r
6 ,pme
2 f4 %) 4 6 =-=õ
; p F
z 00 %) CAN
20 Is
0 F
3 ....
Br
h. itzeoz
F talk f, NCI F WON g.floc20
c0z14 07%) * CO2Me (63 F F 1'1
I SocHN''' F 7
9 ow stops) F g Bac
Abbreviations: PMBOH: 4-methoxybenzyl alcohol; DMF: N-N-dimethylformamide;
DBDMH:
1,3-dibromo-5,5- dimethylhydantoin; TPAP: tetrapropylammonium perruthenate;
NMO: N-
methylmorpholine N-oxide; CAN: ceric ammonium nitrite; DMAP: N,N-
dimethylaminopyridine
In embodiments, starting from ((1R,4S)-2-azabicyclo[2.2.11hept-5-en-3-one) (2)
(also known
as the Vince lactam) and following a modification of literature steps such as
the use of PMBOH/HC1,
(See, Qiu, J.; Silverman, R. B. I Med. Chem. 2000, 43, 706-720), (1R,4R,6S,7R)-
7-bromo-2-(4-
me thoxybenzy1)-3-oxo-2-azabicyclo[2.2.11heptan-6-y1 acetate (4) is obtained
on a multi-gram scale
(Scheme 1A): a. PMBOH, HC1, NaH, THF/DMF; b. DBDMH, AcOH. Alcoholysis of the
acetate and
oxidation yields ketone (1R,4R,7R)-7-bromo-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptane-3,6-
dione (5): c. K2CO3, alcohol; d. TPAP, NMO, 4 A MS, CH2C12, over two steps.
The foregoing allows
difluoro-Horner-Wadsworth-Emmons olefination of ketone 5. When 2-
((difluoromethyl)sulfinyl)pyridine (20) (also known as Hu's reagent) is
employed with KOtBu as
base, (1R, 4R, 7R)-7-bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-
azabicyclo [2 .2.11heptan-3 -
one (6) is obtained: e. 2-((difluoromethyl)sulfinyl)pyridine (20) , KOtBu,
DMF, then NH4C1, then
HC1. Alternatively, e. can be tert-BuLi and F2CHP(0)(0Et)2 (See, Pan, Y.; Qiu,
J.; Silverman, R. B.
I Med. Chem. 2003, 46, 5292-5293). The next step is alcoholysis of the lactam
and elimination.
(1R, 4R,7R)-7-Bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptan-3-one
(6) is deprotected to yield (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-
azabicyclo[2.2.11heptan-3-
one (7): f. CAN, MeCN, H20. A small amount of 4-methoxybenzoyl protected
lactam may also be
isolated. Boc protection of the lactam (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-
azabicyclo[2.2.11heptan-3-one) (7) yields (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-
butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one (8): g. Boc20, DMAP, Et3N,
CH2C12. Alcoholysis
of (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-
azabicyclo[2.2.11heptan-3-
one (8) with K2CO3 and alcohol, leads to subsequent elimination of the bromide
to yield methyl or
ethyl (S)-3-((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-
1-carboxylate (9): h.
K2CO3, alcohol, over two steps. Final deprotection in HC1 yields (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1) with no observable
isomerization or
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degradation: i. HC1, dioxane. Although methanol is shown in the above
schematic, it should be
understood that alcohols such as ethanol or propanol may be utilized as an
alcohol.
In embodiments, starting from ((1R,4S)-2-azabicyclo[2.2.1lhept-5-en-3-one) (2)
(Vince
lactam) and following a modification of literature steps (see, Qiu et al.
supra), e.g., use of
PMBOH/HC1, (1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-
azabicyclo[2.2.11heptan-6-y1
acetate (4) is obtained on a multi-gram scale (Scheme 1A): a. PMBOH (1-2
equiv.), HC1, NaH (0.8-
1.5 equiv.) 0-5 C, THF/DMF (0.75-1.5:0.75-1.5) , 4-8 h,; b. DBDMH (0.4-0.8
equiv), AcOH, 15-
30 C, 4-8 h; Methanolysis of the acetate and oxidation yields ketone
(1R,4R,7R)-7-bromo-2-(4-
me thoxybenzy1)-2-azabicyclo[2.2.1]heptane-3,6-dione (5): c. K2CO3 (2-4
equiv), Me0H 0.5-2 h; d.
TPAP (0.001-0.2 equiv), NMO (1.0-3.0 equiv), 4 A MS, CH2C12, 15-25 h, over two
steps. The
foregoing allows difluoro-Horner-Wadsworth-Emmons olefination of ketone 5.
When 2-
((difluoromethyl)sulfinyl)pyridine (20) (Hu's reagent) is employed with KO'Bu
as base, (1R, 4R, 7R)-
7-bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-
one (6) is
obtained, e.g., infusion of base over 15 minutes to 2 hours with NH4C1/6 M HC1
quench: e. 2-
((difluoromethyl)sulfinyl)pyridine (20) (1.0-1.5 equiv), KO'Bu (1.25-1.75
equiv), DMF, -80--40 C,
15-60 min, then NH4C1, then HC1, then 15-30 C, then 40-80 C, 1 h.
Alternatively, e. can be tert-BuLi
and F2CHP(0)(0Et)2 (See, Pan, Y. et al. supra). The next step is methanolysis
of the lactam and
elimination. (1R,4R,7R)-7-Bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.1]heptan-3-one (6) is deprotected to yield (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7): f. CAN (2-4 equiv),
MeCN, H20, -10
to10 C, 0.75-2 h. A small amount of 4-methoxybenzoyl protected lactam may also
be isolated. Boc
protection of the lactam (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-
azabicyclo[2.2.11heptan-3-one)
(7) yields (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-
azabicyclo[2.2.1]heptan-3-one (8): g. Boc20 (1.0-1.5 equiv), DMAP ( 0.01-0.5
equiv), Et3N (1.0-2.0
equiv), CH2C12, 0.5-2.0 h. Methanolysis of (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-
butoxycarbony1)-2-azabicyclo[2.2.1lheptan-3-one (8) with K2CO3 and methanol,
leads to subsequent
elimination of the bromide to yield methyl (S)-3-((tert-butoxycarbonyl)amino)-
4-
(difluoromethylene)cyclopent-1-ene-1-carboxylate (9): h. K2CO3 (2-4 equiv),
Me0H, 4-8 h, over two
steps. Final deprotection at 70-90 C in 6M HC1 yields (S)-3-amino-4-
(difluoromethylenyl)cyclopent-
1-ene-1-carboxylic acid (1) with no observable isomerization or degradation:
i. HC1 (6 M), dioxane,
70-90 C, 1-3 h.
In embodiments, starting from ((1R,4S)-2-azabicyclo[2.2.1lhept-5-en-3-one) (2)
(Vince
lactam) and following a modification of literature steps such as the use of
PMBOH/HC1, (See, Qiu, J.,
eta al. supra), (1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-
azabicyclo[2.2.11heptan-6-y1
acetate (4) is obtained on a multi-gram scale (Scheme 1A): a. PMBOH (1.5
equiv.), HC1, NaH (1.1
equiv.) 0 C, THF/DMF (1:1) , 6 h, 73%; b. DBDMH (0.6 equiv), AcOH, 23 C, 6 h,
90%;
Methanolysis of the acetate and oxidation yields ketone (1R,4R,7R)-7-bromo-2-
(4-methoxybenzy1)-2-
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azabicyclo[2.2.11heptane-3,6-dione (5): c. K2CO3 (3 equiv), Me0H 1 h; d. TPAP
(0.01 equiv), NMO
(2.0 equiv), 4 A MS, CH2C12, 18 h, 52% over two steps. The foregoing provides
a modality to run
these steps on multi-gram scale allowing difluoro-Horner-Wadsworth-Emmons
olefination of ketone
5. When 2-((difluoromethyl)sulfinyl)pyridine (20) (Hu's reagent) is employed
with KO'Bu as base,
employing Hu's reported conditions (see, Zhao, Y.; Huang, W.; Zhu, L.; Hu, J.
Org. Lett. 2010, 12,
1444-1447), (1R, 4R,7 R)- 7-bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptan-3-one (6) is obtained in small amounts (<10% yield).
Slow infusion of base
over 30 minutes with NH4C1/6 M HC1 quench dramatically increases the yield to
45%. Prolonging
infusion of base to one hour increases the yield to 58%. See, Table 1, below.
The reaction was not
greatly affected by scale, allowing scale up to 3.5 g of (1R,4R,7R)-7-bromo-2-
(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptane-3,6-dione (5) with no decrease in yield: e. 2-
((difluoromethyl)sulfinyl)pyridine (20) (1.2 equiv), KO'Bu (1.5 equiv), DMF, -
60 C, 30 min, then
NH4C1, then 6 M HC1, then 23 C, then 60 C, 1 h. Alternatively, e. can be
tert-BuLi and
F2CHP(0)(0Et)2 (See, Pan, et al., supra). The next step is methanolysis of the
lactam and elimination.
(1R, 4R, 7R)-7-Bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-azabicyclo [2
.2.11heptan-3 -one
(6) is deprotected to yield (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-
azabicyclo[2.2.11heptan-3-
one (7) in 80% yield: f. CAN (3 equiv), MeCN, H20, 0 C, 1 h. A small amount of
4-methoxybenzoyl
protected lactam may also be isolated. Boc protection of the lactam (1R,4R,7R)-
7-bromo-6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one) (7) yields (1R,4R,7R)-7-
bromo-6-
(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one
(8): g. Boc20 (1.2
equiv), DMAP ( 0.1 equiv), Et3N (1.5 equiv), CH2C12, 1 h. Methanolysis of
(1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11hep tan-3-one
(8) with K2CO3 and
methanol, leads to subsequent elimination of the bromide to yield methyl (S)-3-
((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-l-ene-l-carboxylate (9):
h. K2CO3 (3 equiv),
Me0H, 6 h, 52% over two steps. Final deprotection at 80 C in 6M HC1 yields (S)-
3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1) in 97% yield with no
observable
isomerization or degradation: i. HC1 (6 M), dioxane, 80 C, 2 h. Overall, the
yield from Vince Lactam
(2) to S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid (1)
is 8.1%.
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Table 1. Optimization of Fluorination'
F
11/41-
2 0 F
Br
0 2, quench, -60 c'e
430 T.--)60 C
o DMF f."
FMB 6 FMB
6
base
scale
entry (g) addition quench (time)b yield`
method
dropwise
1 0.05 6 M HCI (5 min)
over 5 min
dropwise
2 0.05 6 M HCI ( h) 9%
over 5 min
dropwise a. sat. NH4CI (1 h)
3 0.05 15%
over 5 min b. 6 M HCI (I min)
infusion over a. sat. NH4CI( h)
4 0.13 45%
3o min b. 6 M HCI (2 min)
infusion over a. sat. NH4CI( 1 h)
58%
6o min b. 6 M HCI (2 min)
infusion over a. sat. NH4CI( 1 h)
6 3.5 5o%
90 min b. 6 M HCI (2 min)
a Conditions: 5 (1 equiv), zo (1.2 equiv), DMF (0.3 M) -6o C,
then KCY13u (1.5 equiv) in DMF (0.5 M), then quench at -6o C,
then 23 C, then 6o C for 1 h; b time before quenching
solution was added; isolated yield after chromatography.
In embodiments, without wishing to be bound by any theory, the following is a
proposed
mechanism for fluorination of (1R,4R,7R)-7-Bromo-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptane-3,6-dione (5):
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KO'Su 1 Br
0 --_,, 0 F ,,,- ====-.."-A -60 0 C p F
...,,. 0
0
N
' F pma 0 F Th210 PMS
20 6
_ 11'
¨
B:t Etil
0
F '
0 MB
0.-- ).c..0 FMB
/141: C NH
1\\L" 23 .-.<--4 22
_ ¨
H.1 -fitrc
Y
larli
0 Br
OH -0
HO + N /-
.................................. , IN¨ 4
N
F}.....i...,.. ,
FMB
,11.4-ri7/ 24 F 6
As shown above, multiple intermediates form during the course of the
fluorination reaction.
Intermediate 21, which forms first, rearranges via cyclic intermediate 22 to
form sulfonate 23, which
is then protonated, triggering elimination and formation of the olefin (see
above). If the reaction were
quenched at -60 C with 6 M HC1 five minutes after the addition of KM3u to a
mixture of 5 and 20,
then only 21 was observed by LC/MS (entry 1, Table 1). Addition of KO13u
followed by a 6 M HC1
quench at -60 C (entry 2, Table 1), and subsequent heating at 60 C for 1 h
provided 6 in 9% yield
along with starting material and intermediate 21. Quenching after 1 hour with
a saturated NH4 Cl
solution, followed by 6 M HC1, slightly improved the yield (entry 3, Table 1).
Slow infusion of base
with a syringe pump over 30 min with an NH4C1/6 M HC1 quench dramatically
increased the yield to
45%. Prolonging infusion of base to one hour increased the yield to 58%.
In embodiments, synthesis of (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-
1-
carboxylic acid (1) is shown in Scheme 2.
13
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Scheme 2
kt0,0, Br
'
1 CT
K2C4.
=
THF ---?-
Ac0,,4.1õ.
14., Me0H
O `ems
. v -40 to BOV MOH
2 PA 3 82% 86% 2$ =
67% 84% 76%
,
Br, Brsõ Occ20
TP /It"
Br Hu
AP 0 Fleadiart _,...7.µ40 CAN
F-1-1- %mil WON /Hz() H
Clizat
0' WB r r
$3%
70% 70%
.,
rvroirt\iõ., WOK water 'N' CC.3211 (11) ,L."-W28
WO Btx1HV
HV 142te
f a 73% 19 -70% . 1 H.C4 salt
Racemic synthesis yield (%) is provided over the (s)-isomer yield (%).
Abbreviations: LiHMDS: lithium bis(trimethylsily1) amide; THF:
tetrahydrofuran; PMB: 4-
methoxybenzyl; DBDMH: 1,3 -dibromo-5,5- dimethylhydantoin; TPAP:
tetrapropylammonium
perruthenate; CAN: ceric ammonium nitrite; DMAP: N,N-dimethylaminopyridine
According to Scheme 2, a process for preparing (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1) or salt thereof is
provided which includes
converting ((1R,45)-2-azabicyclo[2.2.11hept-5 -en-3 -one) (2) to (1R,4S)-2-(4-
methoxybenzy1)-2-
azabicyclo[2.2. 11hept-5 -en-3 -one (3). (1R,45)-2-(4-Methoxybenzy1)-2-
azabicyclo[2.2.11hept-5 -en-3 -
one (3) is converted to (1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-
azabicyclo[2.2.11heptan-6-y1 acetate (4). (1R,4R,6S,7R)-7-Bromo-2-(4-
methoxybenzy1)-3-oxo-2-
azabicyclo[2.2.11heptan-6-y1 acetate (4) is converted to (1R,4R,65,7R)-7-bromo-
6-hydroxy-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.1]heptan-3-one (25). (1R,4R,65,7R)-7-bromo-6-
hydroxy-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-one (25) is converted to (1R,4R,7R)-
7 -bromo-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.1]heptane-3,6-dione (5). (1R,4R,7R)-7-Bromo-2-
(4-
methoxybenzy1)-2-azabicyclo[2.2.1]heptane-3,6-dione (5) is converted to
(1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(4-methoxybenzy1)-2-azabicyclo[2.2.1]heptan-3-one (6).
(1R, 4R,7R)-7-Bromo-
6-(difluoromethylene) -2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-one (6)
is converted to
(1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-azabicyclo[2.2.1]heptan-3-one (7).
(1R,4R,7R)-7 -
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Bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7) is converted
to (1R,4R,7R)-7-
bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-
3-one (8).
(1R,4R,7 R)-7 -Bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-
azabicyclo [2.2. 11heptan-3 -one
(8) is converted to (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-
carboxylic acid (19). (S)-34(Tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-
carboxylic acid (19) is converted to (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic
acid (1).
In embodiments, the following compounds are provided:
(3)
(1R,45)-2-(4-Methoxybenzy1)-2-azabicyclop.2.11hept-5-en-3-one (3)
Br
AcOT
tPMB (4)
(1R,4R,6S,7R)-7-Bromo-2-(4-methoxybenzy1)-3-oxo-2-azabicyclo[2.2.11heptan-6-y1
acetate (4)
Br
N,
PMB (25)
(1R,4R,6S,7R)-7-bromo-6-hydroxy-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptan-
3-one (25)
Br
µPMB ( 5)
(1R,4R,7 R)-7 -Bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione
(5)
F
PiVIB
(6)
(1R,4R,7R)-7-Bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptan-3-one (6)
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r 0
F ¨NH
(7)
(1R,4R,7R)-7-Bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7)
Br
Boc
F 15 (8)
(1R,4R,7 R)-7 -Bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-
azabicyclo[2.2.11heptan-3-one
(8)
\
(9)
Methyl (S)-3-((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-
1-carboxylate (9)
F
CO2H
/ BocHil (19)
(S)-3-((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-
carboxylic acid (19)
In embodiments, compositions including (1R,45)-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11hept-5-en-3-one (3) or a salt thereof are provided herein.
Such compositions may
include reaction mixtures such as those described herein. Compositions
including (1R,45)-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.11hept-5-en-3-one (3) or a salt thereof may
include polar solvents,
e.g., aqueous, methanol, ethanol, DMF, acetic acid, etc. or non-polar
solvents, e.g., diethyl ether,
hexane, dichloromethane, ethyl acetate, etc. Compositions including (1R,45)-2-
(4-methoxybenzy1)-2-
azabicyclo[2.2.11hept-5-en-3-one (3) or a pharmaceutically acceptable salt
thereof can be
pharmaceutical compositions. Compositions can include (1R,45)-2-(4-
methoxybenzy1)-2-
azabicyclo[2.2.11hept-5-en-3-one (3) or a salt thereof in amounts of from
0.0001 mg to 50 mg or
more. For example, a pharmaceutical composition may include (1R,45)-2-(4-
methoxybenzy1)-2-
azabicyclo[2.2.11hept-5-en-3-one (3) or a pharmaceutically acceptable salt
thereof in amounts greater
than 0.0001 mg, e.g., from 0.0001 mg to 0.0002 mg, 0.0001 mg to 0.0003 mg,
0.0002 mg to 0.0003
mg, 0.0003 mg to 0.0004 mg, 0.0004 mg to 0.0005 mg, 0.0005 mg to 0.0006 mg,
0.0006 mg to
0.0007 mg, 0.0007 mg to 0.0008 mg, 0.0008 mg to 0.0009 mg, 0.0009 mg to 0.001
mg, 0.001 mg to
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0.002 mg, 0.002 mg to 0.003 mg, 0.003 mg to 0.004 mg, 0.004 mg to 0.005 mg,
0.005 mg to 0.006
mg, 0.006 mg to 0.007 mg, 0.007 mg to 0.008 mg, 0.008 mg to 0.009 mg, 0.009 mg
to 0/.01 mg, 0.01
mg to 0.02 mg, 0.02 mg to 0.03 mg, 0.03 mg to 0.04 mg, 0.04 mg to 0.05 mg,
0.05 mg to 0.06 mg,
0.06 mg to 0.07 mg, 0.07 mg to 0.08 mg, 0.08 mg to 0.09 mg, 0.09 mg to 0.1 mg,
0.1 mg to 0.2 mg,
0.2 mg to 0.3 mg, 0.3 mg to 0.4 mg, 0.4 mg to 0.5 mg, 0.5 mg to 0.6 mg, 0.6 mg
to 0.7 mg, 0.7 mg to
0.8 mg, 0.8 mg to 0.9 mg, 0.9 mg to 1.0 mg, 1.0 mg to 2.0 mg, 2.0 mg to 3.0
mg, 3.0 mg to 4.0 mg,
4.0 mg to 5.0 mg, 5.0 mg to 6.0 mg, 6.0 mg to 7.0 mg, 7.0 mg to 8.0 mg, 8.0 mg
to 9.0 mg, or 9.0 mg
to 10 mg.
In embodiments, compositions including (1R,4R,6S,7R)-7-bromo-2-(4-
methoxybenzy1)-3-
oxo-2-azabicyclo[2.2.11heptan-6-y1 acetate (4) or a salt thereof are provided
herein. Such
compositions may include reaction mixtures such as those described herein.
Compositions including
(1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-azabicyclop.2.11heptan-6-y1
acetate (4) or a
salt thereof may include polar solvents, e.g., aqueous, methanol, ethanol,
DMF, acetic acid, etc. or
non-polar solvents, e.g., diethyl ether, hexane, dichloromethane, ethyl
acetate, etc. Compositions
including (1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-
azabicyclop.2.11heptan-6-y1 acetate
(4) or a pharmaceutically acceptable salt thereof can be pharmaceutical
compositions. Compositions
can include (1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-
azabicyclo[2.2.11heptan-6-y1
acetate (4) or a salt thereof in amounts of from 0.0001 mg to 50 mg or more.
For example, a
pharmaceutical composition may include (1R,4R,6S,7R)-7-bromo-2-(4-
methoxybenzy1)-3-oxo-2-
azabicyclo[2.2.11heptan-6-y1 acetate (4) or a pharmaceutically acceptable salt
thereof in amounts
greater than 0.0001 mg, e.g., from 0.0001 mg to 0.0002 mg, 0.0001 mg to 0.0003
mg, 0.0002 mg to
0.0003 mg, 0.0003 mg to 0.0004 mg, 0.0004 mg to 0.0005 mg, 0.0005 mg to 0.0006
mg, 0.0006 mg
to 0.0007 mg, 0.0007 mg to 0.0008 mg, 0.0008 mg to 0.0009 mg, 0.0009 mg to
0.001 mg, 0.001 mg
to 0.002 mg, 0.002 mg to 0.003 mg, 0.003 mg to 0.004 mg, 0.004 mg to 0.005 mg,
0.005 mg to 0.006
mg, 0.006 mg to 0.007 mg, 0.007 mg to 0.008 mg, 0.008 mg to 0.009 mg, 0.009 mg
to 0.01 mg, 0.01
mg to 0.02 mg, 0.02 mg to 0.03 mg, 0.03 mg to 0.04 mg, 0.04 mg to 0.05 mg,
0.05 mg to 0.06 mg,
0.06 mg to 0.07 mg, 0.07 mg to 0.08 mg, 0.08 mg to 0.09 mg, 0.09 mg to 0.1 mg,
0.1 mg to 0.2 mg,
0.2 mg to 0.3 mg, 0.3 mg to 0.4 mg, 0.4 mg to 0.5 mg, 0.5 mg to 0.6 mg, 0.6 mg
to 0.7 mg, 0.7 mg to
0.8 mg, 0.8 mg to 0.9 mg, 0.9 mg to 1.0 mg, 1.0 mg to 2.0 mg, 2.0 mg to 3.0
mg, 3.0 mg to 4.0 mg,
4.0 mg to 5.0 mg, 5.0 mg to 6.0 mg, 6.0 mg to 7.0 mg, 7.0 mg to 8.0 mg, 8.0 mg
to 9.0 mg, or 9.0 mg
to 10 mg.
In embodiments, compositions including (1R,4R,6S,7R)-7-bromo-6-hydroxy-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-one (25) or a salt thereof are
provided herein. Such
compositions may include reaction mixtures such as those described herein.
Compositions including
(1R,4R,65,7R)-7-bromo-6-hydroxy-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptan-
3-one (25) or a
salt thereof may include polar solvents, e.g., aqueous, methanol, ethanol,
DMF, acetic acid, etc. or
non-polar solvents, e.g., diethyl ether, hexane, dichloromethane, ethyl
acetate, etc. Compositions
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including (1R,4R,6S,7R)-7-bromo-6-hydroxy-2-(4-methoxybenzy1)-2-
azabicyclo112.2.11heptan-3-one
(25) or a pharmaceutically acceptable salt thereof can be pharmaceutical
compositions. Compositions
can include (1R,4R,6S,7R)-7-bromo-6-hydroxy-2-(4-methoxybenzy1)-2-
azabicyclo112.2.11heptan-3-
one (25) or a salt thereof in amounts of from 0.0001 mg to 50 mg or more. For
example, a
pharmaceutical composition may include (1R,4R,6S,7R)-7-bromo-6-hydroxy-2-(4-
methoxybenzy1)-2-
azabicyclo[2.2.11heptan-3-one (25) or a pharmaceutically acceptable salt
thereof in amounts greater
than 0.0001 mg, e.g., from 0.0001 mg to 0.0002 mg, 0.0001 mg to 0.0003 mg,
0.0002 mg to 0.0003
mg, 0.0003 mg to 0.0004 mg, 0.0004 mg to 0.0005 mg, 0.0005 mg to 0.0006 mg,
0.0006 mg to
0.0007 mg, 0.0007 mg to 0.0008 mg, 0.0008 mg to 0.0009 mg, 0.0009 mg to 0.001
mg, 0.001 mg to
0.002 mg, 0.002 mg to 0.003 mg, 0.003 mg to 0.004 mg, 0.004 mg to 0.005 mg,
0.005 mg to 0.006
mg, 0.006 mg to 0.007 mg, 0.007 mg to 0.008 mg, 0.008 mg to 0.009 mg, 0.009 mg
to 0.01 mg, 0.01
mg to 0.02 mg, 0.02 mg to 0.03 mg, 0.03 mg to 0.04 mg, 0.04 mg to 0.05 mg,
0.05 mg to 0.06 mg,
0.06 mg to 0.07 mg, 0.07 mg to 0.08 mg, 0.08 mg to 0.09 mg, 0.09 mg to 0.1 mg,
0.1 mg to 0.2 mg,
0.2 mg to 0.3 mg, 0.3 mg to 0.4 mg, 0.4 mg to 0.5 mg, 0.5 mg to 0.6 mg, 0.6 mg
to 0.7 mg, 0.7 mg to
0.8 mg, 0.8 mg to 0.9 mg, 0.9 mg to 1.0 mg, 1.0 mg to 2.0 mg, 2.0 mg to 3.0
mg, 3.0 mg to 4.0 mg,
4.0 mg to 5.0 mg, 5.0 mg to 6.0 mg, 6.0 mg to 7.0 mg, 7.0 mg to 8.0 mg, 8.0 mg
to 9.0 mg, or 9.0 mg
to 10 mg.
In embodiments, compositions including (1R,4R,7R)-7-bromo-2-(4-methoxybenzy1)-
2-
azabicyclo[2.2.11heptane-3,6-dione (5) or a salt thereof are provided herein.
Such compositions may
include reaction mixtures such as those described herein. Compositions
including (1R,4R,7R)-7-
bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione (5) or a salt
thereof may include
polar solvents, e.g., aqueous, methanol, ethanol, DMF, acetic acid, etc. or
non-polar solvents, e.g.,
diethyl ether, hexane, dichloromethane, ethyl acetate, etc. Compositions
including (1R,4R,7R)-7-
bromo-2-(4-methoxybenzy1)-2-azabicyclo112.2.11heptane-3,6-dione (5) or a
pharmaceutically
acceptable salt thereof can be pharmaceutical compositions. Compositions can
include (1R,4R,7R)-7-
bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione (5) or a salt
thereof in amounts of
from 0.0001 mg to 50 mg or more. For example, a pharmaceutical composition may
include
(1R,4R,7R)-7-bromo-2-(4-methoxybenzy1)-2-azabicyclo112.2.11heptane-3,6-dione
(5) or a
pharmaceutically acceptable salt thereof in amounts greater than 0.0001 mg,
e.g., from 0.0001 mg to
0.0002 mg, 0.0001 mg to 0.0003 mg, 0.0002 mg to 0.0003 mg, 0.0003 mg to 0.0004
mg, 0.0004 mg
to 0.0005 mg, 0.0005 mg to 0.0006 mg, 0.0006 mg to 0.0007 mg, 0.0007 mg to
0.0008 mg, 0.0008
mg to 0.0009 mg, 0.0009 mg to 0.001 mg, 0.001 mg to 0.002 mg, 0.002 mg to
0.003 mg, 0.003 mg to
0.004 mg, 0.004 mg to 0.005 mg, 0.005 mg to 0.006 mg, 0.006 mg to 0.007 mg,
0.007 mg to 0.008
mg, 0.008 mg to 0.009 mg, 0.009 mg to 0.01 mg, 0.01 mg to 0.02 mg, 0.02 mg to
0.03 mg, 0.03 mg to
0.04 mg, 0.04 mg to 0.05 mg, 0.05 mg to 0.06 mg, 0.06 mg to 0.07 mg, 0.07 mg
to 0.08 mg, 0.08 mg
to 0.09 mg, 0.09 mg to 0.1 mg, 0.1 mg to 0.2 mg, 0.2 mg to 0.3 mg, 0.3 mg to
0.4 mg, 0.4 mg to 0.5
mg, 0.5 mg to 0.6 mg, 0.6 mg to 0.7 mg, 0.7 mg to 0.8 mg, 0.8 mg to 0.9 mg,
0.9 mg to 1.0 mg, 1.0
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mg to 2.0 mg, 2.0 mg to 3.0 mg, 3.0 mg to 4.0 mg, 4.0 mg to 5.0 mg, 5.0 mg to
6.0 mg, 6.0 mg to 7.0
mg, 7.0 mg to 8.0 mg, 8.0 mg to 9.0 mg, or 9.0 mg to 10 mg.
In embodiments, compositions including (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-one (6) or a salt thereof are
provided herein. Such
compositions may include reaction mixtures such as those described herein.
Compositions including
(1R, 4R, 7R)-7-bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptan-3-one
(6) or a salt thereof may include polar solvents, e.g., aqueous, methanol,
ethanol, DMF, acetic acid,
etc. or non-polar solvents, e.g., diethyl ether, hexane, dichloromethane,
ethyl acetate, etc.
Compositions including (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-(4-
methoxybenzy1)-2-
azabicyclo[2.2.11heptan-3-one (6) or a pharmaceutically acceptable salt
thereof can be pharmaceutical
compositions. Compositions can include (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-one (6) or a salt thereof in
amounts of from 0.0001 mg
to 50 mg or more. For example, a pharmaceutical composition may include
(1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-one (6) or
a pharmaceutically
acceptable salt thereof in amounts greater than 0.0001 mg, e.g., from 0.0001
mg to 0.0002 mg, 0.0001
mg to 0.0003 mg, 0.0002 mg to 0.0003 mg, 0.0003 mg to 0.0004 mg, 0.0004 mg to
0.0005 mg,
0.0005 mg to 0.0006 mg, 0.0006 mg to 0.0007 mg, 0.0007 mg to 0.0008 mg, 0.0008
mg to 0.0009
mg, 0.0009 mg to 0.001 mg, 0.001 mg to 0.002 mg, 0.002 mg to 0.003 mg, 0.003
mg to 0.004 mg,
0.004 mg to 0.005 mg, 0.005 mg to 0.006 mg, 0.006 mg to 0.007 mg, 0.007 mg to
0.008 mg, 0.008
mg to 0.009 mg, 0.009 mg to 0.01 mg, 0.01 mg to 0.02 mg, 0.02 mg to 0.03 mg,
0.03 mg to 0.04 mg,
0.04 mg to 0.05 mg, 0.05 mg to 0.06 mg, 0.06 mg to 0.07 mg, 0.07 mg to 0.08
mg, 0.08 mg to 0.09
mg, 0.09 mg to 0.1 mg, 0.1 mg to 0.2 mg, 0.2 mg to 0.3 mg, 0.3 mg to 0.4 mg,
0.4 mg to 0.5 mg, 0.5
mg to 0.6 mg, 0.6 mg to 0.7 mg, 0.7 mg to 0.8 mg, 0.8 mg to 0.9 mg, 0.9 mg to
1.0 mg, 1.0 mg to 2.0
mg, 2.0 mg to 3.0 mg, 3.0 mg to 4.0 mg, 4.0 mg to 5.0 mg, 5.0 mg to 6.0 mg,
6.0 mg to 7.0 mg, 7.0
mg to 8.0 mg, 8.0 mg to 9.0 mg, or 9.0 mg to 10 mg.
In embodiments, compositions (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-
azabicyclo[2.2.11heptan-3-one (7) or a salt thereof are provided herein. Such
compositions may
include reaction mixtures such as those described herein. Compositions
including (1R,4R,7R)-7-
bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7) or a salt
thereof may include polar
solvents, e.g., aqueous, methanol, ethanol, DMF, acetic acid, etc. or non-
polar solvents, e.g., diethyl
ether, hexane, dichloromethane, ethyl acetate, etc. Compositions including
(1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7) or a pharmaceutically
acceptable salt thereof
can be pharmaceutical compositions. Compositions can include (1R,4R,7R)-7-
bromo-6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7) or a salt thereof in
amounts of from 0.0001
mg to 50 mg or more. For example, a pharmaceutical composition may include
(1R,4R,7R)-7-bromo-
6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7) or a
pharmaceutically acceptable salt
thereof in amounts greater than 0.0001 mg, e.g., from 0.0001 mg to 0.0002 mg,
0.0001 mg to 0.0003
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mg, 0.0002 mg to 0.0003 mg, 0.0003 mg to 0.0004 mg, 0.0004 mg to 0.0005 mg,
0.0005 mg to
0.0006 mg, 0.0006 mg to 0.0007 mg, 0.0007 mg to 0.0008 mg, 0.0008 mg to 0.0009
mg, 0.0009 mg
to 0.001 mg, 0.001 mg to 0.002 mg, 0.002 mg to 0.003 mg, 0.003 mg to 0.004 mg,
0.004 mg to 0.005
mg, 0.005 mg to 0.006 mg, 0.006 mg to 0.007 mg, 0.007 mg to 0.008 mg, 0.008 mg
to 0.009 mg,
0.009 mg to 0.01 mg, 0.01 mg to 0.02 mg, 0.02 mg to 0.03 mg, 0.03 mg to 0.04
mg, 0.04 mg to 0.05
mg, 0.05 mg to 0.06 mg, 0.06 mg to 0.07 mg, 0.07 mg to 0.08 mg, 0.08 mg to
0.09 mg, 0.09 mg to 0.1
mg, 0.1 mg to 0.2 mg, 0.2 mg to 0.3 mg, 0.3 mg to 0.4 mg, 0.4 mg to 0.5 mg,
0.5 mg to 0.6 mg, 0.6
mg to 0.7 mg, 0.7 mg to 0.8 mg, 0.8 mg to 0.9 mg, 0.9 mg to 1.0 mg, 1.0 mg to
2.0 mg, 2.0 mg to 3.0
mg, 3.0 mg to 4.0 mg, 4.0 mg to 5.0 mg, 5.0 mg to 6.0 mg, 6.0 mg to 7.0 mg,
7.0 mg to 8.0 mg, 8.0
mg to 9.0 mg, or 9.0 mg to 10 mg.
In embodiments, compositions including (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-
butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one (8) or a salt thereof are
provided herein. Such
compositions may include reaction mixtures such as those described herein.
Compositions including
(1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-
azabicyclo[2.2.11heptan-3-one
(8) or a salt thereof may include polar solvents, e.g., aqueous, methanol,
ethanol, DMF, acetic acid,
etc. or non-polar solvents, e.g., diethyl ether, hexane, dichloromethane,
ethyl acetate, etc.
Compositions including (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-(tert-
butoxycarbony1)-2-
azabicyclo[2.2.11heptan-3-one (8) or a pharmaceutically acceptable salt
thereof can be pharmaceutical
compositions. Compositions can include (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-
butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one (8) or a salt thereof in
amounts of from 0.0001 mg
to 50 mg or more. For example, a pharmaceutical composition can include
(1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one
(8) or a
pharmaceutically acceptable salt thereof in amounts greater than 0.0001 mg,
e.g., from 0.0001 mg to
0.0002 mg, 0.0001 mg to 0.0003 mg, 0.0002 mg to 0.0003 mg, 0.0003 mg to 0.0004
mg, 0.0004 mg
to 0.0005 mg, 0.0005 mg to 0.0006 mg, 0.0006 mg to 0.0007 mg, 0.0007 mg to
0.0008 mg, 0.0008
mg to 0.0009 mg, 0.0009 mg to 0.001 mg, 0.001 mg to 0.002 mg, 0.002 mg to
0.003 mg, 0.003 mg to
0.004 mg, 0.004 mg to 0.005 mg, 0.005 mg to 0.006 mg, 0.006 mg to 0.007 mg,
0.007 mg to 0.008
mg, 0.008 mg to 0.009 mg, 0.009 mg to 0.01 mg, 0.01 mg to 0.02 mg, 0.02 mg to
0.03 mg, 0.03 mg to
0.04 mg, 0.04 mg to 0.05 mg, 0.05 mg to 0.06 mg, 0.06 mg to 0.07 mg, 0.07 mg
to 0.08 mg, 0.08 mg
to 0.09 mg, 0.09 mg to 0.1 mg, 0.1 mg to 0.2 mg, 0.2 mg to 0.3 mg, 0.3 mg to
0.4 mg, 0.4 mg to 0.5
mg, 0.5 mg to 0.6 mg, 0.6 mg to 0.7 mg, 0.7 mg to 0.8 mg, 0.8 mg to 0.9 mg,
0.9 mg to 1.0 mg, 1.0
mg to 2.0 mg, 2.0 mg to 3.0 mg, 3.0 mg to 4.0 mg, 4.0 mg to 5.0 mg, 5.0 mg to
6.0 mg, 6.0 mg to 7.0
mg, 7.0 mg to 8.0 mg, 8.0 mg to 9.0 mg, or 9.0 mg to 10 mg.
In embodiments, compositions including methyl (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylate (9) or a salt thereof are
provided herein. Such
compositions may include reaction mixtures such as those described herein.
Compositions including
m ethyl (S)-3-((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-
ene-1-carboxylate (9)
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or a salt thereof may include polar solvents, e.g., aqueous, methanol,
ethanol, DMF, acetic acid, etc.
or non-polar solvents, e.g., diethyl ether, hexane, dichloromethane, ethyl
acetate, etc. Compositions
including methyl (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-
carboxylate (9) or a pharmaceutically acceptable salt thereof can be
pharmaceutical compositions.
Compositions can include methyl (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylate (9) or a salt thereof in
amounts of from 0.0001 mg
to 50 mg or more. For example, a pharmaceutical composition may include methyl
(S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-l-ene-l-carboxylate (9)
or a
pharmaceutically acceptable salt thereof in amounts greater than 0.0001 mg,
e.g., from 0.0001 mg to
0.0002 mg, 0.0001 mg to 0.0003 mg, 0.0002 mg to 0.0003 mg, 0.0003 mg to 0.0004
mg, 0.0004 mg
to 0.0005 mg, 0.0005 mg to 0.0006 mg, 0.0006 mg to 0.0007 mg, 0.0007 mg to
0.0008 mg, 0.0008
mg to 0.0009 mg, 0.0009 mg to 0.001 mg, 0.001 mg to 0.002 mg, 0.002 mg to
0.003 mg, 0.003 mg to
0.004 mg, 0.004 mg to 0.005 mg, 0.005 mg to 0.006 mg, 0.006 mg to 0.007 mg,
0.007 mg to 0.008
mg, 0.008 mg to 0.009 mg, 0.009 mg to 0.01 mg, 0.01 mg to 0.02 mg, 0.02 mg to
0.03 mg, 0.03 mg to
0.04 mg, 0.04 mg to 0.05 mg, 0.05 mg to 0.06 mg, 0.06 mg to 0.07 mg, 0.07 mg
to 0.08 mg, 0.08 mg
to 0.09 mg, 0.09 mg to 0.1 mg, 0.1 mg to 0.2 mg, 0.2 mg to 0.3 mg, 0.3 mg to
0.4 mg, 0.4 mg to 0.5
mg, 0.5 mg to 0.6 mg, 0.6 mg to 0.7 mg, 0.7 mg to 0.8 mg, 0.8 mg to 0.9 mg,
0.9 mg to 1.0 mg, 1.0
mg to 2.0 mg, 2.0 mg to 3.0 mg, 3.0 mg to 4.0 mg, 4.0 mg to 5.0 mg, 5.0 mg to
6.0 mg, 6.0 mg to 7.0
mg, 7.0 mg to 8.0 mg, 8.0 mg to 9.0 mg, or 9.0 mg to 10 mg.
In embodiments, compositions including (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylic acid (19) or a salt thereof
are provided herein. Such
compositions may include reaction mixtures such as those described herein.
Compositions including
(S)-3-((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-
carboxylic acid (19) or a
salt thereof may include polar solvents, e.g., aqueous, methanol, ethanol,
DMF, acetic acid, etc. or
non-polar solvents, e.g., diethyl ether, hexane, dichloromethane, ethyl
acetate, etc. Compositions
including (S)-3-((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-
ene-1-carboxylic
acid (19) or a pharmaceutically acceptable salt thereof can be pharmaceutical
compositions.
Compositions can include (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-
1-carboxylic acid (19) or a salt thereof in amounts of from 0.0001 mg to 50 mg
or more. For example,
a pharmaceutical composition may include (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylic acid (19) or a
pharmaceutically acceptable salt
thereof in amounts greater than 0.0001 mg, e.g., from 0.0001 mg to 0.0002 mg,
0.0001 mg to 0.0003
mg, 0.0002 mg to 0.0003 mg, 0.0003 mg to 0.0004 mg, 0.0004 mg to 0.0005 mg,
0.0005 mg to
0.0006 mg, 0.0006 mg to 0.0007 mg, 0.0007 mg to 0.0008 mg, 0.0008 mg to 0.0009
mg, 0.0009 mg
to 0.001 mg, 0.001 mg to 0.002 mg, 0.002 mg to 0.003 mg, 0.003 mg to 0.004 mg,
0.004 mg to 0.005
mg, 0.005 mg to 0.006 mg, 0.006 mg to 0.007 mg, 0.007 mg to 0.008 mg, 0.008 mg
to 0.009 mg,
0.009 mg to 0.01 mg, 0.01 mg to 0.02 mg, 0.02 mg to 0.03 mg, 0.03 mg to 0.04
mg, 0.04 mg to 0.05
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mg, 0.05 mg to 0.06 mg, 0.06 mg to 0.07 mg, 0.07 mg to 0.08 mg, 0.08 mg to
0.09 mg, 0.09 mg to 0.1
mg, 0.1 mg to 0.2 mg, 0.2 mg to 0.3 mg, 0.3 mg to 0.4 mg, 0.4 mg to 0.5 mg,
0.5 mg to 0.6 mg, 0.6
mg to 0.7 mg, 0.7 mg to 0.8 mg, 0.8 mg to 0.9 mg, 0.9 mg to 1.0 mg, 1.0 mg to
2.0 mg, 2.0 mg to 3.0
mg, 3.0 mg to 4.0 mg, 4.0 mg to 5.0 mg, 5.0 mg to 6.0 mg, 6.0 mg to 7.0 mg,
7.0 mg to 8.0 mg, 8.0
mg to 9.0 mg, or 9.0 mg to 10 mg.
In embodiments, synthesis of (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-
1-
carboxylic acid (1) is shown in Scheme 3 starting with ethyl-cyclopent-3-ene-
carboxylate (10).
Scheme 3
CO2Et 002Et CO2Et
CO2Et
0 _____________________________________________________________
*Bac 0 1µ11-1800 14-1B00
11 la 13
CO2Et 02E1 CO2E1,
. - . .. . ...
'1.1µ111Boa F .N1-11300 F 1012
14 16
As depicted in Scheme 3, a process for preparing (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1) is provided which
includes converting
ethyl-cyclopent-3-ene-carboxylate (10) to (3R,4S)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-
(hydroxy)cyclopentane carboxylate (11). (3R,4S)-Ethy1-3-((tert-
butoxycarbonyl)amino)-4-
(hydroxy)cyclopentane carboxylate (11) is converted to (3R)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-
oxo-cyclopentane carboxylate (12). (3R)-Ethyl-3-((tert-butoxycarbonyl)amino)-4-
oxo-cyclopentane
carboxylate is converted to ethyl (95)-9-(tert-butoxycarbonylamino)-1,4-dioxa-
7-
spiro[4.41nonanecarboxylate (13). Ethyl (95)-9-(tert-butoxycarbonylamino)-1,4-
dioxa-7-
spiro[4.41nonanecarboxylate (13) is converted to ethyl (S)-9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-
spiro[4.41nonene-7-carboxylate (14). Ethyl (S)-9-(tert-butoxycarbonylamino)-
1,4-dioxa-7-
spiro[4.41nonene-7-carboxylate (14) is converted to ethyl (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-l-ene-l-carboxylate (15). Ethyl (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylate (15) is converted to (S)-3-
amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1).
In embodiments, synthesis of (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-
1-
carboxylic acid (1) is shown in Scheme 3A.
22
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WO 2019/227093 PCT/US2019/034140
Scheme 3A
,,.., Sharptess amino CO2Et CO2Et Cop
kou2at horoxylation Oxidation
a
.,S1.. Protection
0 ,
, ..., c,õ0 '-NliBcc
HO. MB= 0 1s4HBoc
ti 12 13
CO2Et CO2Et CO2Et
PhSeBr, Base
deprotection TFA : OCM
H202 0 ,
faimed bY F 1 *Ik114Boo Sat NaHC 3 F i '.141-12
'LA 141-1Boc Horner-Vetig
F 14 reaction F16 1
In embodiments, as shown in Scheme 3A, ethyl-cyclopent-3-ene-carboxylate (10)
(commercially available from Sigma Aldrich, St. Louis, MO) is converted to
(3R,45)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-(hydroxy)cyclopentane carboxylate (11) by Sharpless
aminohydroxylation
and Boc protection. Sharpless aminohydroxylation allows the syn-selective
preparation of 1,2-amino
alcohols by reaction of alkenes with salts of N-halosulfonamides, -amides and -
carbamates using
0s04 as a catalyst. Enantioselectivity is achieved through the addition of
dihydroquinine- and
dihydroquinidine-derived chiral ligands. Oxidation of (3R,45)-ethy1-3-((tert-
butoxycarbonyl)amino)-
4-(hydroxy)cyclopentane carboxylate (11) yields ketone (3R)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-
oxo-cyclopentane carboxylate (12). (3R)-ethyl-3-((tert-butoxycarbonyl)amino)-4-
oxo-cyclopentane
carboxylate (12) is converted to ethyl (95)-9-(tert-butoxycarbonylamino)-1,4-
dioxa-7-
spiro [4 .4]nonanecarboxylate (13). Ethyl (95)-9-(tert-butoxycarbonylamino)-
1,4-dioxa-7-
spiro[4.41nonanecarboxylate (13) is subjected to phenyl selenium bromide, Base
and H202 to yield
Ethyl (S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro [4 .4]nonene-7-
carboxylate (14). Ethyl (5)-
9-(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro [4 .4]nonene-7-carboxylate (14)
is deprotected and
subjected to Horner-Wittig reaction to yield ethyl (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylate (15). The Horner-Wittig
reaction involves the
reaction of aldehydes or ketones with stabilized phosphorus ylides
(phosphonate carbanions) and
leads to olefins with E-selectivity. Ethyl (S)-3-((tert-butoxycarbonyl)amino)-
4-
(difluoromethylene)cyclopent-l-ene-l-carboxylate (15) is converted to (S)-3-
amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1) with trifluoroacetic
acid (TFA),
dichloromethane (DCM) and saturated NaHCO3.
In embodiments, the following compounds are provided:
23
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co2Et
o
Hd õNFIBoc
(3R,45)-ethyl-3-((tert-butoxycarbonyl)amino)-4-(hydroxy)cyclopentane
carboxylate (11)
902Et
O NHBoe
(3R)-ethyl-3-((tert-butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12)
902Et
0 ______________________________
'-NHBoe
(13)
Ethyl (9S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-
spiro[4.4]nonanecarboxylate (13)
902Et
0 _______________________________ .
'NHBoc
(14)
Ethyl (S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro[4.4]nonene-7-
carboxylate (14)
CO2Et
F .--NHBoc
Ethyl (S)-3-((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-
1-carboxylate (15)
In embodiments, compositions including (3R,4S)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-
(hydroxy)cyclopentane carboxylate (11) or a salt thereof are provided herein.
Such compositions may
include reaction mixtures such as those described herein. Compositions (3R,45)-
ethy1-3-((tert-
butoxycarbonyl)amino)-4-(hydroxy)cyclopentane carboxylate (11) or a salt
thereof may include polar
solvents, e.g., aqueous, methanol, ethanol, DMF, acetic acid, etc. or non-
polar solvents, e.g., diethyl
ether, hexane, dichloromethane, ethyl acetate, etc. Compositions including
(3R,45)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-(hydroxy)cyclopentane carboxylate (11) or a
pharmaceutically acceptable
salt thereof can be pharmaceutical compositions. Compositions can include
(3R,4S)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-(hydroxy)cyclopentane carboxylate (11) or a salt
thereof in amounts of
from 0.0001 mg to 50 mg or more. For example, a pharmaceutical composition may
include (3R,45)-
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ethyl-3-((tert-butoxycarbonyl)amino)-4-(hydroxy)cyclopentane carboxylate (11)
or a
pharmaceutically acceptable salt thereof in amounts greater than 0.0001 mg,
e.g., from 0.0001 mg to
0.0002 mg, 0.0001 mg to 0.0003 mg, 0.0002 mg to 0.0003 mg, 0.0003 mg to 0.0004
mg, 0.0004 mg
to 0.0005 mg, 0.0005 mg to 0.0006 mg, 0.0006 mg to 0.0007 mg, 0.0007 mg to
0.0008 mg, 0.0008
mg to 0.0009 mg, 0.0009 mg to 0.001 mg, 0.001 mg to 0.002 mg, 0.002 mg to
0.003 mg, 0.003 mg to
0.004 mg, 0.004 mg to 0.005 mg, 0.005 mg to 0.006 mg, 0.006 mg to 0.007 mg,
0.007 mg to 0.008
mg, 0.008 mg to 0.009 mg, 0.009 mg to 0.01 mg, 0.01 mg to 0.02 mg, 0.02 mg to
0.03 mg, 0.03 mg to
0.04 mg, 0.04 mg to 0.05 mg, 0.05 mg to 0.06 mg, 0.06 mg to 0.07 mg, 0.07 mg
to 0.08 mg, 0.08 mg
to 0.09 mg, 0.09 mg to 0.1 mg, 0.1 mg to 0.2 mg, 0.2 mg to 0.3 mg, 0.3 mg to
0.4 mg, 0.4 mg to 0.5
mg, 0.5 mg to 0.6 mg, 0.6 mg to 0.7 mg, 0.7 mg to 0.8 mg, 0.8 mg to 0.9 mg,
0.9 mg to 1.0 mg, 1.0
mg to 2.0 mg, 2.0 mg to 3.0 mg, 3.0 mg to 4.0 mg, 4.0 mg to 5.0 mg, 5.0 mg to
6.0 mg, 6.0 mg to 7.0
mg, 7.0 mg to 8.0 mg, 8.0 mg to 9.0 mg, or 9.0 mg to 10 mg.
In embodiments, compositions including (3R)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-oxo-
cyclopentane carboxylate (12) or a salt thereof are provided herein. Such
compositions may include
reaction mixtures such as those described herein. Compositions including (3R)-
ethy1-3-((tert-
butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12) or a salt thereof
may include polar
solvents, e.g., aqueous, methanol, ethanol, DMF, acetic acid, etc. or non-
polar solvents, e.g., diethyl
ether, hexane, dichloromethane, ethyl acetate, etc. Compositions including
(3R)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12) or a
pharmaceutically acceptable salt
thereof can be pharmaceutical compositions. Compositions can include (3R)-
ethy1-3-((tert-
butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12) or a salt thereof in
amounts of from
0.0001 mg to 50 mg or more. For example, a pharmaceutical composition can
include ((3R)-ethy1-3-
((tert-butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12) or a
pharmaceutically acceptable
salt thereof in amounts greater than 0.0001 mg, e.g., from 0.0001 mg to 0.0002
mg, 0.0001 mg to
0.0003 mg, 0.0002 mg to 0.0003 mg, 0.0003 mg to 0.0004 mg, 0.0004 mg to 0.0005
mg, 0.0005 mg
to 0.0006 mg, 0.0006 mg to 0.0007 mg, 0.0007 mg to 0.0008 mg, 0.0008 mg to
0.0009 mg, 0.0009
mg to 0.001 mg, 0.001 mg to 0.002 mg, 0.002 mg to 0.003 mg, 0.003 mg to 0.004
mg, 0.004 mg to
0.005 mg, 0.005 mg to 0.006 mg, 0.006 mg to 0.007 mg, 0.007 mg to 0.008 mg,
0.008 mg to 0.009
mg, 0.009 mg to 0.01 mg, 0.01 mg to 0.02 mg, 0.02 mg to 0.03 mg, 0.03 mg to
0.04 mg, 0.04 mg to
0.05 mg, 0.05 mg to 0.06 mg, 0.06 mg to 0.07 mg, 0.07 mg to 0.08 mg, 0.08 mg
to 0.09 mg, 0.09 mg
to 0.1 mg, 0.1 mg to 0.2 mg, 0.2 mg to 0.3 mg, 0.3 mg to 0.4 mg, 0.4 mg to 0.5
mg, 0.5 mg to 0.6 mg,
0.6 mg to 0.7 mg, 0.7 mg to 0.8 mg, 0.8 mg to 0.9 mg, 0.9 mg to 1.0 mg, 1.0 mg
to 2.0 mg, 2.0 mg to
3.0 mg, 3.0 mg to 4.0 mg, 4.0 mg to 5.0 mg, 5.0 mg to 6.0 mg, 6.0 mg to 7.0
mg, 7.0 mg to 8.0 mg,
8.0 mg to 9.0 mg, or 9.0 mg to 10 mg.
In embodiments, compositions including ethyl (95)-9-(tert-butoxycarbonylamino)-
1,4-dioxa-
7-spiro[4.41nonanecarboxylate (13) or a salt thereof are provided herein. Such
compositions may
include reaction mixtures such as those described herein. Compositions
including including ethyl
CA 03101472 2020-11-24
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(9S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro[4.4]nonanecarboxylate (13)
or a salt thereof
may include polar solvents, e.g., aqueous, methanol, ethanol, DMF, acetic
acid, etc. or non-polar
solvents, e.g., diethyl ether, hexane, dichloromethane, ethyl acetate, etc.
Compositions including
including ethyl (9S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-
spiro[4.4]nonanecarboxylate (13) or a
pharmaceutically acceptable salt thereof can be pharmaceutical compositions.
Compositions can
include including ethyl (9S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-
spiro[4.4]nonanecarboxylate
(13) or a salt thereof in amounts of from 0.0001 mg to 50 mg or more. For
example, a pharmaceutical
composition can include including ethyl (9S)-9-(tert-butoxycarbonylamino)-1,4-
dioxa-7-
spiro[4.4]nonanecarboxylate (13) or a pharmaceutically acceptable salt thereof
in amounts greater
than 0.0001 mg, e.g., from 0.0001 mg to 0.0002 mg, 0.0001 mg to 0.0003 mg,
0.0002 mg to 0.0003
mg, 0.0003 mg to 0.0004 mg, 0.0004 mg to 0.0005 mg, 0.0005 mg to 0.0006 mg,
0.0006 mg to
0.0007 mg, 0.0007 mg to 0.0008 mg, 0.0008 mg to 0.0009 mg, 0.0009 mg to 0.001
mg, 0.001 mg to
0.002 mg, 0.002 mg to 0.003 mg, 0.003 mg to 0.004 mg, 0.004 mg to 0.005 mg,
0.005 mg to 0.006
mg, 0.006 mg to 0.007 mg, 0.007 mg to 0.008 mg, 0.008 mg to 0.009 mg, 0.009 mg
to 0.01 mg, 0.01
mg to 0.02 mg, 0.02 mg to 0.03 mg, 0.03 mg to 0.04 mg, 0.04 mg to 0.05 mg,
0.05 mg to 0.06 mg,
0.06 mg to 0.07 mg, 0.07 mg to 0.08 mg, 0.08 mg to 0.09 mg, 0.09 mg to 0.1 mg,
0.1 mg to 0.2 mg,
0.2 mg to 0.3 mg, 0.3 mg to 0.4 mg, 0.4 mg to 0.5 mg, 0.5 mg to 0.6 mg, 0.6 mg
to 0.7 mg, 0.7 mg to
0.8 mg, 0.8 mg to 0.9 mg, 0.9 mg to 1.0 mg, 1.0 mg to 2.0 mg, 2.0 mg to 3.0
mg, 3.0 mg to 4.0 mg,
4.0 mg to 5.0 mg, 5.0 mg to 6.0 mg, 6.0 mg to 7.0 mg, 7.0 mg to 8.0 mg, 8.0 mg
to 9.0 mg, or 9.0 mg
to 10 mg.
In embodiments, compositions including ethyl (S)-9-(tert-butoxycarbonylamino)-
1,4-dioxa-7-
spiro[4.4]nonene-7-carboxylate (14) or a salt thereof are provided herein.
Such compositions may
include reaction mixtures such as those described herein. Compositions
including ethyl (S)-9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-spiro[4.4]nonene-7-carboxylate (14) or a salt
thereof may include
polar solvents, e.g., aqueous, methanol, ethanol, DMF, acetic acid, etc. or
non-polar solvents, e.g.,
diethyl ether, hexane, dichloromethane, ethyl acetate, etc. Compositions
including ethyl (S)-9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-5pir0[4.4]nonene-7-carboxylate (14) or a
pharmaceutically
acceptable salt thereof can be pharmaceutical compositions. Compositions can
include ethyl (S)-9-
(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro[4.4]nonene-7-carboxylate (14) or
a salt thereof in
amounts of from 0.0001 mg to 50 mg or more. For example, a pharmaceutical
composition can
include ethyl (S)-9-(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro[4.4]nonene-7-
carboxylate (14) or a
pharmaceutically acceptable salt thereof in amounts greater than 0.0001 mg,
e.g., from 0.0001 mg to
0.0002 mg, 0.0001 mg to 0.0003 mg, 0.0002 mg to 0.0003 mg, 0.0003 mg to 0.0004
mg, 0.0004 mg
to 0.0005 mg, 0.0005 mg to 0.0006 mg, 0.0006 mg to 0.0007 mg, 0.0007 mg to
0.0008 mg, 0.0008
mg to 0.0009 mg, 0.0009 mg to 0.001 mg, 0.001 mg to 0.002 mg, 0.002 mg to
0.003 mg, 0.003 mg to
0.004 mg, 0.004 mg to 0.005 mg, 0.005 mg to 0.006 mg, 0.006 mg to 0.007 mg,
0.007 mg to 0.008
mg, 0.008 mg to 0.009 mg, 0.009 mg to 0.01 mg, 0.01 mg to 0.02 mg, 0.02 mg to
0.03 mg, 0.03 mg to
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0.04 mg, 0.04 mg to 0.05 mg, 0.05 mg to 0.06 mg, 0.06 mg to 0.07 mg, 0.07 mg
to 0.08 mg, 0.08 mg
to 0.09 mg, 0.09 mg to 0.1 mg, 0.1 mg to 0.2 mg, 0.2 mg to 0.3 mg, 0.3 mg to
0.4 mg, 0.4 mg to 0.5
mg, 0.5 mg to 0.6 mg, 0.6 mg to 0.7 mg, 0.7 mg to 0.8 mg, 0.8 mg to 0.9 mg,
0.9 mg to 1.0 mg, 1.0
mg to 2.0 mg, 2.0 mg to 3.0 mg, 3.0 mg to 4.0 mg, 4.0 mg to 5.0 mg, 5.0 mg to
6.0 mg, 6.0 mg to 7.0
mg, 7.0 mg to 8.0 mg, 8.0 mg to 9.0 mg, or 9.0 mg to 10 mg.
In embodiments, compositions including ethyl (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylate (15) or a salt thereof are
provided herein. Such
compositions may include reaction mixtures such as those described herein.
Compositions including
ethyl (S)-3-((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-
1-carboxylate (15) or
a salt thereof may include polar solvents, e.g., aqueous, methanol, ethanol,
DMF, acetic acid, etc. or
non-polar solvents, e.g., diethyl ether, hexane, dichloromethane, ethyl
acetate, etc. Compositions
including ethyl (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-
carboxylate (15) or a pharmaceutically acceptable salt thereof can be
pharmaceutical compositions.
Compositions can include ethyl (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-
1-ene-1-carboxylate (15) or a salt thereof in amounts of from 0.0001 mg to 50
mg or more. For
example, a pharmaceutical composition can include ethyl (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylate (15) or a pharmaceutically
acceptable salt thereof
in amounts greater than 0.0001 mg, e.g., from 0.0001 mg to 0.0002 mg, 0.0001
mg to 0.0003 mg,
0.0002 mg to 0.0003 mg, 0.0003 mg to 0.0004 mg, 0.0004 mg to 0.0005 mg, 0.0005
mg to 0.0006
mg, 0.0006 mg to 0.0007 mg, 0.0007 mg to 0.0008 mg, 0.0008 mg to 0.0009 mg,
0.0009 mg to 0.001
mg, 0.001 mg to 0.002 mg, 0.002 mg to 0.003 mg, 0.003 mg to 0.004 mg, 0.004 mg
to 0.005 mg,
0.005 mg to 0.006 mg, 0.006 mg to 0.007 mg, 0.007 mg to 0.008 mg, 0.008 mg to
0.009 mg, 0.009
mg to 0.01 mg, 0.01 mg to 0.02 mg, 0.02 mg to 0.03 mg, 0.03 mg to 0.04 mg,
0.04 mg to 0.05 mg,
0.05 mg to 0.06 mg, 0.06 mg to 0.07 mg, 0.07 mg to 0.08 mg, 0.08 mg to 0.09
mg, 0.09 mg to 0.1 mg,
0.1 mg to 0.2 mg, 0.2 mg to 0.3 mg, 0.3 mg to 0.4 mg, 0.4 mg to 0.5 mg, 0.5 mg
to 0.6 mg, 0.6 mg to
0.7 mg, 0.7 mg to 0.8 mg, 0.8 mg to 0.9 mg, 0.9 mg to 1.0 mg, 1.0 mg to 2.0
mg, 2.0 mg to 3.0 mg,
3.0 mg to 4.0 mg, 4.0 mg to 5.0 mg, 5.0 mg to 6.0 mg, 6.0 mg to 7.0 mg, 7.0 mg
to 8.0 mg, 8.0 mg to
9.0 mg, or 9.0 mg to 10 mg.
In embodiments, pharmaceutical compositions may include (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically
acceptable salt thereof
in an amount of, e.g., about 0.001 to 500 mg, 0.01 to 500 mg, 0.01 to 450 mg,
0.01 to 300 mg, 0.01 to
250 mg, 0.01 to 200 mg, 0.01 to 175 mg, 0.01 to 150 mg, 0.01 to 125 mg, 0.01
to 100 mg, 0.01 to 75
mg, 0.01 to 50 mg, 0.01 to 30 mg, 0.01 to 25 mg, 0.01 to 20 mg, 0.01 to 15 mg,
0.01 to 10 mg, 0.01 to
mg, 0.01 to lmg, 0.025 to 500 mg, 0.025 to 450 mg, 0.025 to 300 mg, 0.025 to
250 mg, 0.025 to
200 mg, 0.025 to 175 mg, 0.025 to 150 mg, 0.025 to 125 mg, 0.025 to 100 mg,
0.025 to 75 mg, 0.025
to 50 mg, 0.025 to 30 mg, 0.025 to 25 mg, 0.025 to 20 mg, 0.025 to 15 mg,
0.025 to 10 mg, 0.025 to 5
mg, 0.025 to lmg, 0.05 to 500 mg, 0.05 to 450 mg, 0.05 to 300 mg, 0.05 to 250
mg, 0.05 to 200 mg,
27
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0.05 to 175 mg, 0.05 to 150 mg, 0.05 to 125 mg, 0.05 to 100 mg, 0.05 to 75 mg,
0.05 to 50 mg, 0.05
to 30 mg, 0.05 to 25 mg, 0.05 to 20 mg, 0.05 to 15 mg, 0.05 to 10 mg, 0.05 to
5 mg, 0.05 to lmg,
0.075 to 500 mg, 0.075 to 450 mg, 0.075 to 300 mg, 0.075 to 250 mg, 0.075 to
200 mg, 0.075 to 175
mg, 0.075 to 150 mg, 0.075 to 125 mg, 0.075 to 100 mg, 0.075 to 75 mg, 0.075
to 50 mg, 0.075 to 30
mg, 0.075 to 25 mg, 0.075 to 20 mg, 0.075 to 15 mg, 0.075 to 10 mg, 0.075 to 5
mg, 0.075 to lmg,
0.1 to 500 mg, 0.1 to 450 mg, 0.1 to 300 mg, 0.1 to 250 mg, 0.1 to 200 mg, 0.1
to 175 mg, 0.1 to 150
mg, 0.1 to 125 mg, 0.1 to 100 mg, 0.1 to 75 mg, 0.1 to 50 mg, 0.1 to 30 mg,
0.1 to 25 mg, 0.1 to 20
mg, 0.1 to 15 mg, 0.1 to 10 mg, 0.1 to 5 mg, 0.1 to lmg, 0.25 to 500 mg, 0.25
to 450 mg, 0.25 to 300
mg, 0.25 to 250 mg, 0.25 to 200 mg, 0.25 to 175 mg, 0.25 to 150 mg, 0.25 to
125 mg, 0.25 to 100 mg,
0.25 to 75 mg, 0.25 to 50 mg, 0.25 to 30 mg, 0.25 to 25 mg, 0.25 to 20 mg,
0.25 to 15 mg, 0.25 to 10
mg, 0.25 to 5 mg, 0.25 to lmg, 0.05 to 500 mg, 0.5 to 450 mg, 0.5 to 300 mg,
0.5 to 250 mg, 0.5 to
200 mg, 0.5 to 175 mg, 0.5 to 150 mg, 0.5 to 125 mg, 0.5 to 100 mg, 0.5 to 75
mg, 0.5 to 50 mg, 0.5
to 30 mg, 0.5 to 25 mg, 0.5 to 20 mg, 0.5 to 15 mg, 0.5 to 10 mg, 0.5 to 5 mg,
0.5 to lmg, 1 to 500
mg, 1 to 450 mg, 1 to 300 mg, 1 to 250 mg, 1 to 200 mg, 1 to 175 mg, 1 to 150
mg, 1 to 125 mg, 1 to
100 mg, 1 to 75 mg, 1 to 50 mg, 1 to 30 mg, 1 to 25 mg, 1 to 20 mg, 1 to 15
mg, 1 to 10 mg, 1 to 5
mg, 1 to 4 mg, 1 to 3 mg, 1 to 2 mg, 2 to 500 mg, 2 to 450 mg, 2 to 300 mg, 2
to 250 mg, 2 to 200 mg,
2 to 175 mg, 2 to 150 mg, 2 to 125 mg, 2 to 100 mg, 2 to 75 mg, 2 to 50 mg, 2
to 30 mg, 2 to 25 mg, 2
to 20 mg, 2 to 15 mg, 2 to 10 mg, 2 to 5 mg, 3 to 500 mg, 3 to 450 mg, 3 to
300 mg, 3 to 250 mg, 3 to
200 mg, 3 to 175 mg, 3 to 150 mg, 3 to 125 mg, 3 to 100 mg, 3 to 75 mg, 3 to
50 mg, 3 to 30 mg, 3 to
25 mg, 3 to 20 mg, 3 to 15 mg, 3 to 10 mg, 3 to 5 mg, 4 to 500 mg, 4 to 450
mg, 4 to 300 mg, 4 to 250
mg, 4 to 200 mg, 4 to 175 mg, 4 to 150 mg, 4 to 125 mg, 4 to 100 mg, 4 to 75
mg, 4 to 50 mg, 4 to 30
mg, 4 to 25 mg, 4 to 20 mg, 4 to 15 mg, 4 to 10 mg, 4 to 5 mg, 5 to 500 mg, 5
to 450 mg, 5 to 300 mg,
to 250 mg, 5 to 200 mg, 5 to 175 mg, 5 to 150 mg, 5 to 125 mg, 5 to 100 mg, 5
to 75 mg, 5 to 50
mg, 5 to 30 mg, 5 to 25 mg, 5 to 20 mg, 5 to 15 mg, 5 to 10 mg, 10 to 500 mg,
10 to 450 mg, 10 to
300 mg, 10 to 250 mg, 10 to 200 mg, 10 to 175 mg, 10 to 150 mg, 10 to 125 mg,
10 to 100 mg, 10 to
75 mg, 10 to 50 mg, 10 to 30 mg, 10 to 25 mg, 10 to 20 mg, 10 to 15 mg, 15 to
500 mg, 15 to 450 mg,
to 300 mg, 15 to 250 mg, 15 to 200 mg, 15 to 175 mg, 15 to 150 mg, 15 to 125
mg, 15 to 100 mg,
15 to 75 mg, 15 to 50 mg, 15 to 30 mg, 15 to 25 mg, 15 to 20 mg, 20 to 500 mg,
20 to 450 mg, 20 to
300 mg, 20 to 250 mg, 20 to 200 mg, 20 to 175 mg, 20 to 150 mg, 20 to 125 mg,
20 to 100 mg, 20 to
75 mg, 20 to 50 mg, 20 to 30 mg, 20 to 25 mg, 25 to 500 mg, 25 to 450 mg, 25
to 300 mg, 25 to 250
mg, 25 to 200 mg, 25 to 175 mg, 25 to 150 mg, 25 to 125 mg, 25 to 100 mg, 25
to 80 mg, 25 to 75
mg, 25 to 50 mg, 25 to 30 mg, 30 to 500 mg, 30 to 450 mg, 30 to 300 mg, 30 to
250 mg, 30 to 200
mg, 30 to 175 mg, 30 to 150 mg, 30 to 125 mg, 30 to 100 mg, 30 to 75 mg, 30 to
50 mg, 40 to 500
mg, 40 to 450 mg, 40 to 400 mg, 40 to 250 mg, 40 to 200 mg, 40 to 175 mg, 40
to 150 mg, 40 to 125
mg, 40 to 100 mg, 40 to 75 mg, 40 to 50 mg, 50 to 500 mg, 50 to 450 mg, 50 to
300 mg, 50 to 250
mg, 50 to 200 mg, 50 to 175 mg, 50 to 150 mg, 50 to 125 mg, 50 to 100 mg, 50
to 75 mg, 75 to 500
mg, 75 to 450 mg, 75 to 300 mg, 75 to 250 mg, 75 to 200 mg, 75 to 175 mg, 75
to 150 mg, 75 to 125
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mg, 75 to 100 mg, 100 to 500 mg, 100 to 450 mg, 100 to 300 mg, 100 to 250 mg,
100 to 200 mg, 100
to 175 mg, 100 to 150 mg, 100 to 125 mg, 125 to 500 mg, 125 to 450 mg, 125 to
300 mg, 125 to 250
mg, 125 to 200 mg, 125 to 175 mg, 125 to 150 mg, 150 to 500 mg, 150 to 450 mg,
150 to 300 mg,
150 to 250 mg, 150 to 200 mg, 200 to 500 mg, 200 to 450 mg, 200 to 300 mg, 200
to 250 mg, 250 to
500 mg, 250 to 450 mg, 250 to 300 mg, 300 to 500 mg, 300 to 450 mg, 300 to 400
mg, 300 to 350
mg, 350 to 500 mg, 350 to 450 mg, 350 to 400 mg, 400 to 500 mg, 400 to 450 mg,
with 0.01 mg,
0.025 mg, 0.05 mg, 0.075 mg, 0.1 mg, 0.25 mg, 0.5 mg, 0.75 mg, 1 mg, 2 mg, 2.5
mg, 3 mg, 4 mg, 5
mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 30 mg, 35
mg, 40 mg, 45 mg,
50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg,
125 mg, 150 mg
175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400
mg, 425 mg, 450
mg, 475 mg, and 500 mg being examples.
In embodiments, pharmaceutical compositions including (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1) and one or more of
(1R,4S)-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.11hept-5-en-3-one (3), (1R,4R,6S,7R)-7-bromo-2-
(4-
methoxybenzy1)-3-oxo-2-azabicyclo[2.2.11heptan-6-y1 acetate (4), (1R,4R,7R)-7-
bromo-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione (5), (1R,4R,7 R)- 7-bromo-
6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (6), (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7), (1R,4R,7R)-7-bromo-6-
(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one
(8), or methyl (S)-3-
((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-
carboxylate (9) are provided.
In embodiments, pharmaceutical compositions including (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid (1) and one or more of
ethyl-cyclopent-3-ene-
carboxylate (10), (3R,4S)-ethyl-3-((tert-butoxycarbonyl)amino)-4-
(hydroxy)cyclopentane carboxylate
(11), (3R)-ethyl-3-((tert-butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate
(12), ethyl (9S)-9-
(tert-butoxycarbonylamino)-1,4-dioxa-7-spiro[4.41nonanecarboxylate (13), ethyl
(S)-9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-spiro[4.41nonene-7-carboxylate (14), or ethyl
(S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-carboxylate (15),
are provided.
In embodiments, (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic
acid may
be provided as an acid addition salt, a zwitter ion hydrate, zwitter ion
anhydrate, hydrochloride or
hydrobromide salt, or in the form of the zwitter ion monohydrate. Acid
addition salts, include but are
not limited to, maleic, fumaric, benzoic, ascorbic, succinic, oxalic, bis-
methylenesalicylic,
methanesulfonic, ethane-disulfonic, acetic, propionic, tartaric, salicylic,
citric, gluconic, lactic, malic,
mandelic, cinnamic, citraconic, aspartic, stearic, palmitic, itaconic,
glycolic, pantothenic, p-amino-
benzoic, glutamic, benzene sulfonic or theophylline acetic acid addition
salts, as well as the 8-
halotheophyllines, for example 8-bromo-theophylline. In embodiments, inorganic
acid addition salts,
including but not limited to, hydrochloric, hydrobromic, hydroiodic, sulfuric,
sulfamic, phosphoric or
nitric acid addition salts may be used.
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In embodiments, (1R,45)-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11hept-5-en-3-one
(3),
(1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-azabicyclo[2.2.11heptan-6-y1
acetate (4),
(1R,4R,7R)-7-bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione
(5), (1R,4R,7R)-7-
bromo-6-(difluoromethylene)-2-azabicyclo [2.2.11heptan-3 -one (6), (1R,4R,7R)-
7-bromo-6-
(difluoromethylene)-2-azabicyclo [2.2.11heptan-3 -one (7), (1R,4R,7R)-7-bromo-
6-
(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one
(8), or methyl (S)-3-
((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-
carboxylate (9) may be
provided as an acid addition salt, a zwitter ion hydrate, zwitter ion
anhydrate, hydrochloride or
hydrobromide salt, or in the form of the zwitter ion monohydrate. Acid
addition salts, include but are
not limited to, maleic, fumaric, benzoic, ascorbic, succinic, oxalic, bis-
methylenesalicylic,
methanesulfonic, ethane-disulfonic, acetic, propionic, tartaric, salicylic,
citric, gluconic, lactic, malic,
mandelic, cinnamic, citraconic, aspartic, stearic, palmitic, itaconic,
glycolic, pantothenic, p-amino-
benzoic, glutamic, benzene sulfonic or theophylline acetic acid addition
salts, as well as the 8-
halotheophyllines, for example 8-bromo-theophylline. In embodiments, inorganic
acid addition salts,
including but not limited to, hydrochloric, hydrobromic, hydroiodic, sulfuric,
sulfamic, phosphoric or
nitric acid addition salts may be used.
In embodiments, ethyl-cyclopent-3-ene-carboxylate (10), (3R,45)-ethy1-3-((tert-
butoxycarbonyl)amino)-4-(hydroxy)cyclopentane carboxylate (11), (3R)-ethy1-3-
((tert-
butoxycarbonyl)amino)-4-oxo-cyclopentane carboxylate (12), ethyl (9S)-9-(tert-
butoxycarbonylamino)-1,4-dioxa-7-spiro[4.41nonanecarboxylate (13), ethyl (S)-9-
(tert-
butoxycarbonylamino)-1,4-dioxa-7-spiro[4.41nonene-7-carboxylate (14), or ethyl
(S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-carboxylate (15)
may be provided as
an acid addition salt, a zwitter ion hydrate, zwitter ion anhydrate,
hydrochloride or hydrobromide salt,
or in the form of the zwitter ion monohydrate. Acid addition salts, include
but are not limited to,
maleic, fumaric, benzoic, ascorbic, succinic, oxalic, bis-methylenesalicylic,
methanesulfonic, ethane-
disulfonic, acetic, propionic, tartaric, salicylic, citric, gluconic, lactic,
malic, mandelic, cinnamic,
citraconic, aspartic, stearic, palmitic, itaconic, glycolic, pantothenic, p-
amino-benzoic, glutamic,
benzene sulfonic or theophylline acetic acid addition salts, as well as the 8-
halotheophyllines, for
example 8-bromo-theophylline. In embodiments, inorganic acid addition salts,
including but not
limited to, hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfamic,
phosphoric or nitric acid
addition salts may be used.
In embodiments, pharmaceutical compositions include various dosage forms
including
conventional formulations and modified release formulations. Such
pharmaceutical compositions may
be adapted for any suitable route of administration, e.g., oral, rectal,
nasal, ophthalmic, pulmonary,
vaginal, sublingual, transdermal, intravenous, intraarterial, intramuscular,
intraperitoneal and
subcutaneous routes. Suitable dosage forms include tablets, capsules, oral
liquids, ophthalmic drops,
CA 03101472 2020-11-24
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ophthalmic ointments, ophthalmic gels, powders, aerosols, transdermal
modalities such as topical
liquids, patches, creams and ointments, parenteral formulations and
suppositories.
In embodiments, as mentioned previously, pharmaceutical compositions herein
may be
provided with conventional release or modified release profiles.
Pharmaceutical compositions may be
prepared using a pharmaceutically acceptable "carrier" composed of materials
that are considered safe
and effective. The "carrier" includes all components present in the
pharmaceutical formulation other
than the active ingredient or ingredients. The term "carrier" includes, but is
not limited to, diluents,
binders, lubricants, disintegrants, fillers, and coating compositions. Those
with skill in the art are
familiar with such pharmaceutical carriers and methods of compounding
pharmaceutical compositions
using such carriers.
In embodiments, pharmaceutical compositions herein are modified release dosage
forms
which provide modified release profiles. Modified release profiles may exhibit
immediate release,
delayed release, or extended release profiles. Conventional (or unmodified)
release oral dosage forms
such as tablets, capsules, suppositories, syrups, solutions and suspensions
typically release
medications into the mouth, stomach or intestines as the tablet, capsule shell
or suppository dissolves,
or, in the case of syrups, solutions and suspensions, when they are swallowed.
The pattern of drug
release from modified release (MR) dosage forms is deliberately changed from
that of a conventional
dosage form to achieve a desired therapeutic objective and/or better patient
compliance. Types of MR
drug products include orally disintegrating dosage forms (ODDFs) which provide
immediate release,
extended release dosage forms, delayed release dosage forms (e.g., enteric
coated), and pulsatile
release dosage forms.
An ODDF is a solid dosage form containing a medicinal substance or active
ingredient which
disintegrates rapidly, usually within a matter of seconds when placed upon the
tongue. The
disintegration time for ODDFs generally range from one or two seconds to about
a minute. ODDFs
are designed to disintegrate or dissolve rapidly on contact with saliva. This
mode of administration
can be beneficial to people who may have problems swallowing tablets whether
it be from physical
infirmity or psychiatric in nature. Some subjects with an eye disorder may
exhibit such behavior.
ODDF's can provide rapid delivery of medication to the blood stream through
mucosa resulting in a
rapid onset of action. Examples of ODDFs include orally disintegrating
tablets, capsules and rapidly
dissolving films and wafers.
Extended release dosage forms (ERDFs) have extended release profiles and are
those that
allow a reduction in dosing frequency as compared to that presented by a
conventional dosage form,
e.g., a solution or unmodified release dosage form. ERDFs provide a sustained
duration of action of a
drug. Suitable formulations which provide extended release profiles are well-
known in the art. For
example, coated slow release beads or granules ("beads" and "granules" are
used interchangeably
herein) in which any of the compounds described herein are applied to beads,
e.g., confectioners
nonpareil beads, and then coated with conventional release retarding materials
such as waxes, enteric
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coatings and the like. In embodiments, beads can be formed in which any of the
compounds described
herein are mixed with a material to provide a mass from which the compound
leaches out. In
embodiments, the beads may be engineered to provide different rates of release
by varying
characteristics of the coating or mass, e.g., thickness, porosity, using
different materials, etc. Beads
having different rates of release may be combined into a single dosage form to
provide variable or
continuous release. The beads can be contained in capsules or compressed into
tablets.
In embodiments, modified dosage forms herein incorporate delayed release
dosage forms
having delayed release profiles. Delayed release dosage forms can include
delayed release tablets or
delayed release capsules. A delayed release tablet is a solid dosage form
which releases a compound
(or compounds) described herein are at a time other than promptly after
administration. A delayed
release capsule is a solid dosage form in which the drug is enclosed within
either a hard or soft soluble
container made from a suitable form of gelatin, and which releases a drug (or
drugs) at a time other
than promptly after administration. For example, enteric-coated tablets,
capsules, particles and beads
are well-known examples of delayed release dosage forms. Enteric coated
tablets, capsules and
particles and beads pass through the stomach and release the drug in the
intestine. In embodiments, a
delayed release tablet is a solid dosage form containing a conglomerate of
medicinal particles that
releases a drug (or drugs) at a time other than promptly after administration.
In embodiments, the
conglomerate of medicinal particles are covered with a coating which delays
release of the drug. In
embodiments, a delayed release capsule is a solid dosage form containing a
conglomerate of
medicinal particles that releases a drug (or drugs) at a time other than
promptly after administration.
In embodiments, the conglomerate of medicinal particles are covered with a
coating which delays
release of the drug.
Delayed release dosage forms are known to those skilled in the art. For
example, coated
delayed release beads or granules in which any of the compounds described
herein are applied to
beads, e.g., confectioners nonpareil beads, and then coated with conventional
release delaying
materials such as waxes, enteric coatings and the like. In embodiments, beads
can be formed in which
any of the compounds described herein are mixed with a material to provide a
mass from which the
drug leaches out. In embodiments, the beads may be engineered to provide
different rates of release
by varying characteristics of the coating or mass, e.g., thickness, porosity,
using different materials,
etc. In embodiments, enteric coated granules of any of the compounds described
herein can be
contained in an enterically coated capsule or tablet which releases the
granules in the small intestine.
In embodiments, the granules have a coating which remains intact until the
coated granules reach at
least the ileum and thereafter provide a delayed release of the drug in the
colon. Suitable enteric
coating materials are well known in the art, e.g., Eudragit0 coatings such
methacrylic acid and methyl
methacrylate polymers and others. The granules can be contained in capsules or
compressed into
tablets.
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In embodiments, any of the compounds described herein are incorporated into
porous inert
carriers that provide delayed release profiles. In embodiments, the porous
inert carriers incorporate
channels or passages from which the drug diffuses into surrounding fluids. In
embodiments, any of the
compounds described herein are incorporated into an ion-exchange resin to
provide a delayed release
profile. Delayed action may result from a predetermined rate of release of the
drug from the resin when
the drug-resin complex contacts gastrointestinal fluids and the ionic
constituents dissolved therein. In
embodiments, membranes are utilized to control rate of release from drug
containing reservoirs. In
embodiments, liquid preparations may also be utilized to provide a delayed
release profile. For
example, a liquid preparation consisting of solid particles dispersed
throughout a liquid phase in which
the particles are not soluble. The suspension is formulated to allow at least
a reduction in dosing
frequency as compared to that drug presented as a conventional dosage form
(e.g., as a solution or a
prompt drug-releasing, conventional solid dosage form). For example, a
suspension of ion-exchange
resin constituents or microbe ads.
In embodiments, pharmaceutical compositions described herein are suitable for
ophthalmic or
parenteral administration, including, e.g., intramuscular (i.m.), intravenous
(iv.), subcutaneous (s.c.),
intraperitoneal (i.p.), or intrathecal (it). Parenteral or ophthalmic
compositions must be sterile for
administration by injection, infusion, instillation or implantation into the
body and may be packaged
in either single-dose or multi-dose containers. In embodiments, liquid
pharmaceutical compositions
for ophthalmic or parenteral administration to a subject include an active
substance, e.g., any of the
compounds described herein are in any of the respective amounts described
above. In embodiments,
the pharmaceutical compositions for ophthalmic or parenteral administration
are formulated as a total
volume of about, e.g., 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 7.5 m1,10 ml, 20 ml, 25
ml, 50 ml, 100 ml, 200 ml,
250 ml, or 500 ml. In embodiments, the compositions are contained in a bag, a
glass vial, a plastic
vial, or a bottle.
In embodiments, pharmaceutical compositions for ophthalmic or parenteral
administration
include respective amounts described above for any of the compounds described
herein. In
embodiments, pharmaceutical compositions for ophthalmic or parenteral
administration include about
0.0001 mg to about 500 mg of any of the compounds described herein. In
embodiments,
pharmaceutical compositions for ophthalmic or parenteral administration to a
subject may include any
of the compounds described herein, at a respective concentration of about
0.005 mg/ml to about 500
mg/ml. In embodiments, the pharmaceutical composition for ophthalmic or
parenteral administration
includes any of the compounds described herein at a respective concentration
of, e.g., about 0.05
mg/ml to about 50 mg/ml, about 0.1 mg/ml to about 50 mg/ml, about 0.1 mg/ml to
about 10 mg/ml,
about 0.05 mg/ml to about 25 mg/ml, about 0.05 mg/ml to about 10 mg/ml, about
0.05 mg/ml to about
mg/ml, or about 0.05 mg/ml to about 1 mg/ml. In embodiments, the
pharmaceutical composition for
ophthalmic or parenteral administration includes any of the compounds
described herein at a
respective concentration of, e.g., about 0.05 mg/ml to about 15 mg/ml, about
0.5 mg/ml to about 10
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mg/ml, about 0.25 mg/ml to about 5 mg/ml, about 0.5 mg/ml to about 7 mg/ml,
about 1 mg/ml to
about 10 mg/ml, about 5 mg/ml to about 10 mg/ml, or about 5 mg/ml to about 15
mg/ml.
In embodiments, a pharmaceutical composition for ophthalmic or parenteral
administration is
provided wherein the pharmaceutical composition is stable for at least six
months. In embodiments,
the pharmaceutical compositions for ophthalmic or parenteral administration
exhibit no more than
about 5% decrease in active substance, e.g., (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic acid, or a pharmaceutically acceptable salt of thereof, e.g., 3
months or 6 months. In
embodiments, the amount of ( (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-
1-carboxylic acid,
or a pharmaceutically acceptable salt thereof, degrades at no more than about,
e.g., 2.5%, 1%, 0.5% or
0.1%. In embodiments, the degradation is less than about, e.g., 5%, 2.5%, 1%,
0.5%, 0.25%, 0.1%, for
at least six months.
In embodiments, pharmaceutical compositions for ophthalmic or parenteral
administration are
provided wherein the pharmaceutical composition remains soluble. In
embodiments, pharmaceutical
compositions for ophthalmic or parenteral administration are provided that are
stable, soluble, local
site compatible and/or ready-to-use. In embodiments, the pharmaceutical
compositions herein are
ready-to-use for direct administration to a subject in need thereof
The pharmaceutical compositions for ophthalmic or parenteral administration
provided herein
may include one or more excipients, e.g., solvents, solubility enhancers,
suspending agents, buffering
agents, isotonicity agents, stabilizers or antimicrobial preservatives. When
used, the excipients of the
ophthalmic or parenteral compositions will not adversely affect the stability,
bioavailability, safety,
and/or efficacy of any of the compounds described herein used in the
composition. Thus, ophthalmic
or parenteral compositions are provided wherein there is no incompatibility
between any of the
components of the dosage form.
In embodiments, ophthalmic or parenteral compositions including any of the
compounds
described herein include a stabilizing amount of at least one excipient. For
example, excipients may
be selected from the group consisting of buffering agents, solubilizing
agents, tonicity agents,
antioxidants, chelating agents, antimicrobial agents, and preservatives. One
skilled in the art will
appreciate that an excipient may have more than one function and be classified
in one or more defined
group.
In embodiments, ophthalmic or parenteral compositions any of the compounds
described
herein and an excipient wherein the excipient is present at a weight percent
(w/v) of less than about,
e.g., 10%, 5%, 2.5%, 1%, or 0.5%. In embodiments, the excipient is present at
a weight percent
between about, e.g., 1.0% to 10%, 10% to 25%, 15% to 35%, 0.5% to 5%, 0.001%
to 1%, 0.01% to
1%, 0.1% to 1%, or 0.5% to 1%. In embodiments, the excipient is present at a
weight percent between
about, e.g., 0.001% to 1%, 0.01% to 1%, 1.0% to 5%, 10% to 15%, or 1% to 15%.
In embodiments, ophthalmic or parenteral compositions of any of the compounds
described
herein are provided, wherein the pH of the composition is between about 4.0 to
about 8Ø In
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embodiments, the pH of the compositions is between, e.g., about 5.0 to about
8.0, about 6.0 to about
8.0, about 6.5 to about 8Ø In embodiments, the pH of the compositions is
between, e.g., about 6.5 to
about 7.5, about 7.0 to about 7.8, about 7.2 to about 7.8, or about 7.3 to
about 7.6. In embodiments,
the pH of the aqueous solution is, e.g., about 6.8, about 7.0, about 7.2,
about 7.4, about 7.6, about 7.7,
about 7.8, about 8.0, about 8.2, about 8.4, or about 8.6.
In embodiments, pharmaceutical compositions including (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid, or a pharmaceutically
acceptable salt thereof,
provide an in vivo plasma profile having a C. of (S)-3-amino-4-
(difluoromethylenyl)cyclopent-l-
ene- 1-carboxylic acid less than about, e.g., 2000 ng/ml, 1000 ng/ml, 850
ng/ml, 800 ng/ml, 750 ng/ml,
700 ng/ml, 650 ng/ml, 600 ng/ml, 550 ng/ml, 450 ng/ml, 400 ng/ml 350 ng/ml, or
300 ng/ml. In
embodiments, the pharmaceutical composition provides an in vivo plasma profile
having a C. less
than about, e.g., 250 ng/ml, 200 ng/ml 150 ng/ml, or 100 ng/ml.
In embodiments, provided herein are pharmaceutical compositions containing (5)-
3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid, or a pharmaceutically
acceptable salt thereof
and one or more of any of the compounds described herein, wherein the
composition provides a
consistent in vivo plasma profile of (S)-3-amino-4-
(difluoromethylenyl)cyclopent-l-ene-l-carboxylic
acid having a AUC0_. of less than about 900 ngehr/ml.
In embodiments, the Tmax of pharmaceutical compositions containing (S)-3-amino-
4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid, or a pharmaceutically
acceptable salt thereof
and one or more of any of the compounds described herein is less than 3 hours.
In embodiments, the
T. of the pharmaceutical composition is less than 2.5 hours. In embodiments,
the Tmax of the
pharmaceutical composition is less than 2 hours. In embodiments, the T. of the
pharmaceutical
composition is less than 1.5 hours. In embodiments, the Tmax of the
pharmaceutical composition is less
than 1 hour. In embodiments, the T. of the pharmaceutical composition is less
than 0.5 hour. In
embodiments, the T. of the pharmaceutical composition is less than 0.25 hour.
In embodiments, pharmaceutical compositions containing (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid, or a pharmaceutically
acceptable salt thereof
and one or more of any of the compounds described herein provide a dissolution
of at least about 80%
within the first 20 minutes of administration to a subject in need thereof In
embodiments,
pharmaceutical compositions containing (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic acid, or a pharmaceutically acceptable salt thereof and one or more
of any of the
compounds described herein provide a dissolution of at least about, e.g., 85%,
90% or 9 5 % within the
first 20 minutes of administration to a subject in need thereof. In
embodiments, pharmaceutical
compositions containing (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic acid, or a
pharmaceutically acceptable salt thereof and one or more of any of the
compounds described herein
provide a dissolution of at least 80% within the first 10 minutes of
administration to a subject in need
thereof.
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It should be understood that respective amounts of (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid, or a pharmaceutically
acceptable salt thereof
and one or more of any of the compounds described herein are applicable to all
the dosage forms
described herein including conventional dosage forms, modified dosage forms,
as well as the
ophthalmic and parenteral formulations described herein. Those skilled in the
art will determine
appropriate amounts depending on criteria such as dosage form, route of
administration, subject
tolerance, efficacy, therapeutic goal and therapeutic benefit, among other
pharmaceutically acceptable
criteria.
Unless defined otherwise, all technical and scientific terms used herein have
the same
meanings as commonly understood by one of skill in the art to which the
disclosure herein belongs.
The term "about" or "approximately" as used herein means within an acceptable
error range
for the particular value as determined by one of ordinary skill in the art,
which will depend in part on
how the value is measured or determined, i.e., the limitations of the
measurement system. For
example, "about" can mean within 3 or more than 3 standard deviations, per the
practice in the art.
Alternatively, "about" can mean a range of up to 20%, up to 10%, up to 5%,
and/or up to 1% of a
given value. Alternatively, particularly with respect to biological systems or
processes, the term can
mean within an order of magnitude, preferably within 5-fold, and more
preferably within 2-fold, of a
value.
"PK" refers to the pharmacokinetic profile. C. is defined as the highest
plasma drug
concentration estimated during an experiment (ng/ml). T. is defined as the
time when C. is
estimated (min). AUC0_. is the total area under the plasma drug concentration-
time curve, from drug
administration until the drug is eliminated (ng.hr/m1). The area under the
curve is governed by
clearance. Clearance is defined as the volume of blood or plasma that is
totally cleared of its content
of drug per unit time (ml/min).
"Pharmaceutically acceptable" refers to molecular entities and compositions
that are
"generally regarded as safe"-e.g., that are physiologically tolerable and do
not typically produce an
allergic or similar untoward reaction, such as gastric upset and the like,
when administered to a
human. In embodiments, this term refers to molecular entities and compositions
approved by a
regulatory agency of the federal or a state government, as the GRAS list under
section 204(s) and 409
of the Federal Food, Drug and Cosmetic Act, that is subject to premarket
review and approval by the
FDA or similar lists, the U.S. Pharmacopeia or another generally recognized
pharmacopeia for use in
animals, and more particularly in humans.
"Effective amount" or "therapeutically effective amount" means a dosage
sufficient to
alleviate one or more symptoms of a disorder, disease, or condition being
treated.
"Co-administered with", "co-therapy", "in combination with", "a combination
of', "combined
with" or "administered along with" may be used interchangeably and mean that
two or more agents
are administered in the course of therapy. The agents may be administered
together at the same time
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or separately in spaced apart intervals. The agents may be administered in a
single dosage form or in
separate dosage forms.
"Patient in need thereof' includes individuals that have been diagnosed with a
disease,
condition or disorder for which treatment with (S)-3-amino-4-
(difluoromethylenyl)cyclopent-1-ene-1-
carboxylic acid, or a pharmaceutically acceptable salt thereof is indicated.
"Patient" and "subject" are
used interchangeably herein.
EXAMPLES
The Examples provided herein are included solely for augmenting the disclosure
herein and
should not be considered to be limiting in any respect.
Methods In General
(1R,4S)-2-Azabicyclo[2.2.11hept-5-en-3-one (2) was purchased from Acella
Chembio, San
Diego, CA 92121 or AK Scientific, Inc., Union City, CA 94587. 4-Methoxybenzyl
chloride was
purchased from AK Scientific, Inc., Union City, CA 94587.
2-(difluoromethyl)sulfinyl)pyridine (20) was either purchased from Enamine
Chemicals, Monmouth
Jct., NJ 08852, Sigma-Aldrich Corp, St Louis, MO, or synthesized from diethyl
bromodifluoromethylphosphonate and 2-mercaptopyridine. See, Zhou, Q.; Ruffoni,
A.; Gianatassio,
R.; Fujiwara, Y.; Sella, E.; Shabat, D.; Baran, P. S. Angew. Chem. Int. Ed.
2013, 52, 3949-3952. 1,3-
dibromo-5,5-dimethylhydantoin was purchased from Acros Organics division of
Thermo Fisher
Scientific, Waltham, MA 02451. TPAP was purchased from Combi Blocks, Inc. San
Diego, CA
92126. Cerium ammonium nitrate was purchased from Alfa Aesar, Ward Hill, MA
01835. All other
reagents were purchased from Sigma-Aldrich, Fisher Scientific or Acros
Organics and used without
further purification. Anhydrous solvents (THF, CH2C12, DMF) were purified
before use by passing
through a column composed of activated alumina and a supported copper redox
catalyst. Yields refer
to chromatographically and spectroscopically (1H-NMR) homogeneous material.
Analytical thin-layer
chromatography (TLC) was performed using Merck Silica Gel 60 A F-254 precoated
plates (0.25 mm
thickness), and components were visualized by ultraviolet light (254 nm)
and/or ceric ammonium
molybdate stain. Flash column chromatography was performed on a Teledyne
Combiflash Rf Plus
automated flash purification system with various Teledyne cartridges (4-80 g,
40-63 um, 60 A).
Purifications were performed with hexanes and ethyl acetate unless otherwise
noted. III and 13C NMR
spectra were recorded on a Bruker Avance-III NMR spectrometer at 500 MHz and
126 MHz,
respectively, in CDC13 or D20. Chemical shifts were reported in ppm,
multiplicities are indicated by s
= singlet, d = doublet, t = triplet, q = quartet, sep = septet, dd = doublet
of doublet, dt = doublet of
triplet, m = multiplet, br = broad resonance. Coupling constants were reported
in Hz. High resolution
mass spectral data were obtained on an Agilent 6210 LC-TOF spectrometer in the
positive ion mode
using electrospray ionization with an Agilent G1312A HPLC pump and an Agilent
G1367B
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autoinjector at the Integrated Molecular Structure Education and Research
Center (IMSERC),
Northwestern University. Analytical HPLC was performed by using a reserved-
phase Agilent Infinity
1260 HPLC with a Phenomenex Kintex C-18 column (50 x 2.1 mm, 2.6 jtm),
detecting with UV
absorbance at 254 nm.
Example 1
Manufacture of (1R,45)-2-(4-Methoxybenzy1)-2-azabicyclo[2.2.11hept-5-en-3-one
(3)
Method A
4-Methoxybenzyl alcohol (35.80 mL, 0.29 mol, 1.5 equiv) was added dropwise to
concentrated HC1 (300 mL) and stirred for 1 h. Water was added, and the liquid
was extracted (2 x
100 mL) with diethyl ether. The diethyl ether was dried over Na2SO4 and
concentrated to a volume of
about 50 mL. To a 2 L flask, equipped with an addition funnel, was added
(1R,45)-2-
azabicyclo[2.2.11hept-5-en-3-one (21.00 g, 0.19 mol), DMF (600 mL) ,and THF
(600 mL), and the
flask was cooled to 0 C. NaH (8.45 g, 0.21 mol, 1.1 equiv, 60% dispersion in
mineral oil) was added
portionwise. The flask was placed under N2 and stirred for 30 min. The
Et20/PMBC1 solution was
transferred to the addition funnel and was added dropwise at 0 C. The
reaction was stirred for 6 h at
room temperature. Upon completion, THF was removed in vacuo, and diethyl ether
and water were
added. Any solids were filtered, and the layers were separated. The aqueous
layer was extracted (3 x
100 mL) with diethyl ether, and the organic layers were combined and washed
with brine (2 x 200
mL). After drying over Na2SO4 and concentration, a yellow oil was obtained.
The crude oil was
purified by flash chromatography to yield 32.2 g (0.14 mol, 73% yield) of
protected (1R,4S)-2-
azabicyclo[2.2.11hept-5-en-3-one (3). Spectra matched those in the literature.
See, Qiu, J.; Silverman,
R. B. I Med. Chem. 2000, 43, 706-720.
Method B
A dry 2-liter 3-neck round bottom flask was fitted with an overhead stirrer/
Teflon paddle,
500 ml dropping funnel, and a serum cap/Type-K thermocouple /N2 needle. (1R4S)-
(+2-Aza-
bicyclo[2.2.11hept-5-en-3-one, (-)-2 (25 gm, 229 mmol, 1 equiv) was dissolved
in anhydrous THF
(600 ml) in the flask and the dropping funnel charged with the LiHMDS solution
(1M soln. in
toluene). The flask contents were cooled to -40 C and the LiHMDS solution
(252 ml, 252 mmol, 1.2
equiv) was added in drop by drop into the flask keeping the temperature around
-35 to 40 C. The
flask content started to thicken and became a thin gel towards the end of the
base addition. The
reaction mixture was stirred for further 1 hour after the LiHMDS had all been
added, and then the 500
ml dropping funnel was removed and replaced with a smaller 125 ml dropping
funnel charged with 4-
methoxybenzyl chloride (37.2 gm, 240.5 mmol, 1.05 equiv). The halide was added
into the reaction
flask dropwise. When approximately 1/4 of the chloride had been added,
tetrabutylammonium iodide
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powder (1.3 g, 3.44 mmol, 0.015 equiv) was added in a single portion. When all
the 4-methoxybenzyl
chloride was added the flask was warmed to room temperature initially and then
warmed to 60 C in
an heating mantle. The progress of the benzylation reaction was followed over
time by hplc and by
lcms. The reaction was worked up after 2 days at 60 C when the reaction had
reached ¨85%
conversion to lactam (-)(1R,4S)-2-azabicyclo[2.2.11hept-5-en-3-one (3). The
fine solid precipitate in
the reaction solution was removed by filtering through a large 17 cm Buchner
funnel/ filter paper
under vacuum. The residues were washed with THF (200 ml) and also collected
with the original
filtrate and the combined golden yellow filtrates were concentrated (rotary
evaporator/vacuum/ water
bath at 40 C). The residual golden oil was dissolved in Et0Ac (350 ml) and
washed with water (250
m1). After the phase split, the aqueous layer was back extracted with Et0Ac (2
x 100 m1). The Et0Ac
layers were combined and dried over anhydrous Na2SO4 powder, filtered and
concentrated (rotary
evaporator/vacuum/ water bath at 40 C) to a golden yellow liquid. This crude
product was purified
by chromatography on the Biotage with a 340 g silica column and an Et0Ac-
hexanes gradient [15%
(2 CV) 15-75% (8 CV) 100% (3 CV)] with fractions collected in 25 x 150 mm
tubes. The product
containing fractions were combined and concentrated (rotary evaporator/vacuum/
water bath at 40 C)
to a golden yellow liquid which was pumped down on the vacuum line for several
hours.
Method C
NaH (40.3 g, 1.01 mol, 60% purity, 1.1 equiv) and Bu4NI (16.9 g, 45.8 mmol,
0.05 equiv)
were added to THF (300 mL), then a solution of (1R,4S)-2-azabicyclo[2.2.11hept-
5-en-3-one (100.0 g,
916.3 mmol, 1 equiv) in DMF (900 mL) was added dropwise to the mixture at 0 C.
The mixture was
stirred for 0.5 h, and para-methoxybenzyl chloride (215.2 g, 1.37 mol, 186.5
mL, 1.50 equiv) was
added dropwise to the above mixture at 0 C. The mixture was stirred at 25 C
for 1 h. TLC
(petroleum ether: ethyl acetate = 1:1, Rf = 0.20) showed the reactants were
consumed completely.
The mixture was diluted with MTBE (2 L) and water (3.5 L) was added. Any
solids were filtered,
and the layers were separated. The aqueous layer was extracted with MTBE (800
mL x 2) and the
organic layers were combined and washed with brine (1 L x 2). The combined
organic layers were
dried over anhydrous Na2SO4and concentrated to give a residue as a yellow oil.
The residue was
purified by column chromatography on silica gel with petroleum ether: ethyl
acetate (10:1-1:1) to
give a yellow oil. The crude (1R,4S)-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11hept-5-en-3-one (3)
(89.5 g, 390.3 mmol, 42.6% yield) was obtained as a yellow oil, which was used
in the next step
without further purification.
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Example 2
Manufacture of (1R,4R,6S,7R)-7-Bromo-2-(4-methoxybenzy1)-
3-oxo-2-azabicyclo[2.2.11heptan-6-y1 acetate (4)
Method A
To a solution of (1R,45)-2-(4-Methoxybenzy1)-2-azabicyclo[2.2.11hept-5-en-3-
one (3), (10.00
g, 43.62 mmol) in AcOH (110.0 mL) was added 1,3-dibromo-5,5-dimethylhydantoin
(7.48 g, 26.17
mmol, 0.6 equiv). The reaction was stirred for 6 h, and upon completion, water
was added. The
aqueous layer was extracted with diethyl ether (3 x 200 mL), and the organic
layers were combined,
washed with 1 M NaOH, dried over Na2SO4, and concentrated. Purification by
flash chromatography
yielded (1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-
azabicyclo[2.2.11heptan-6-y1 acetate
(4) (14.40 g, 39.25 mmol, 90% yield) as a thick oil. Spectra matched those in
the literature. See, Qiu,
J.; Silverman, R. B. I Med. Chem. 2000, 43, 706-720.
Method B
(1R, 4R)-(-)-2-( 4-Methoxybenzy1)-2-azabicyclo[2.2.1 lhept-5-en-3-one (15.5 g,
67.60 mmol,
1 equiv) was dissolved in glacial acetic acid (125 ml) in a 250 ml round
bottom flask with a stir-bar.
1,3-Dibromo-5,5-di-methylhydantoin (9.7 g, 33.43 mmol, 0.5 equiv) was added to
the acidic solution
in multiple batches in 10 minutes at room temperature. A serum cap/nitrogen
needle was placed in the
neck of the flask and the reaction mixture was stirred overnight at room
temperature. In 20 minutes,
after all the dibromide was added, the initial cloudy yellow solution turned
clear golden yellow. The
progress of the reaction was monitored by hplc and by lcms. After stirring
overnight the reaction
solvent was removed (rotary evaporator/vacuum / water bath at 55 C to afford
a dark golden yellow
liquid that was then dissolved in dichloromethane (200 m1). Water was added
(100 ml), and the
mixture transferred into a 1-L Erlenmeyer flask. 10% Sodium sulfite solution
(75 ml) was added and
stirred vigorously in the conical flask for a 1/2-hour to destroy unreacted
dibromohydantoin. More
water (100 ml) was added and this was followed by 3M sodium hydroxide, added
slowly and swirled
in the conical flask, to bring the aqueous layer to pH= 7 (pH paper). The
flask contents were
transferred into in a 1-L separatory funnel; the conical flask was rinsed with
dichloromethane (100
ml) and the rinse was combined in the funnel. The dichloromethane layer from
the phase split was
saved, the aqueous layer was back extracted with dichloromethane (2 x 100 ml)
and combined with
the original saved extract which were washed with satd. NaHCO3 (250 ml), water
(250 ml) and brine
(250 m1). The organic phase was dried with anhydrous Na2SO4 powder, filtered
and concentrated
(rotary evaporator/vacuum/ water bath at 40 C) to a golden yellow liquid.
This crude product was
purified by chromatography on the Biotage with a 340 g silica column and a
Et0Ac-hexanes gradient
[15% (1 CV) 15-75% (7.5 CV) 100% (3 CV)], fractions collected in 25 x 150 mm
tubes. The fractions
containing the product were combined and concentrated (rotary
evaporator/vacuum/ water bath at 40
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C) to give a golden yellow liquid which was pumped down further on the vacuum
line for several
hours, thus yielding purified (1R,4R,6S,7R)-(+ )-7-bromo-2-( 4-methoxybenzy1)-
3-oxo-2-azabicyclo[
2.2.1 Fheptan-6-y1 acetate (4) (23 g; 82%).
Example 3
Manufacture of (1R,4R,7R)-7-Bromo-2-(4-methoxybenzy1)-
2-azabicyclo[2.2.11heptane-3,6-dione (5)
Method A
(1R,4R,6S,7R)-7-Bromo-2-(4-methoxybenzy1)-3-oxo-2-azabicyclo[2.2.11heptan-6-y1
acetate
(4) (12.8 g, 34.7 mmol) was dissolved in Me0H (270 mL) and K2CO3 (14.40 g,
104.28 mmol, 3.0
equiv) was added. The reaction was stirred for 1 h, filtered, and then
concentrated. Ethyl acetate and
water were added, and the layers were separated. The organic layer was dried
over Na2SO4 and
concentrated to yield an off-white colored solid 41R,4R,6S,7R)-7-Bromo-6-
hydroxy-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.11heptan-3-one) (25), which was used directly
in the next step.
This compound was placed in a 500 mL flask and purged with argon.
Dichloromethane (170
mL) was added followed by 4 A molecular sieves (10 g). TPAP (122.2 mg, 0.35
mmol. 0.01 equiv)
and NMO (8.14 g, 69.52 mmol, 2.0 equiv) were then added, and the reaction
mixture was stirred
overnight. The reaction mixture was then filtered and concentrated to a volume
of 20 mL and loaded
directly onto a flash chromatography column. The resulting yellowish solid can
be recrystallized from
hexanes/ethyl acetate to obtain a white powder (1R,4R,7R)-7-bromo-2-(4-
methoxybenzy1)-2-
azabicyclo[2.2.11heptane-3,6-dione (5) (5.96 g, 18.38 mmol, 52% yield).
Spectra matched those in the
literature. See, Qiu, J.; Silverman, R. B. I Med. Chem. 2000, 43, 706-720.
Method B
(1R,4R,6S,7R)-7-Bromo-2-(4-methoxybenzy1)-3-oxo-2-azabicyclo[2.2.11heptan-6-y1
acetate
(4) (25.5 g, 69.2 mmol) was dissolved in Me0H (300 mL) and K2CO3 (30 g, 0.23
mol, 3 equiv) was
added. The reaction was stirred for 1 h, filtered, and then concentrated.
Ethyl acetate and water were
added, and the layers were separated. The organic layer was dried over Na2SO4
and concentrated to
yield an off-white colored solid ((lR,4R,6S,7R)-7-Bromo-6-hydroxy-2-(4-
methoxybenzy1)-2-
azabicyclo[2.2.11heptan-3-one), which was used directly in the next step.
A three-neck flask was equipped with a vent line to a bubble, dropping funnel
with nitrogen inlet, and
septum. Dichloromethane (160 mL) was added, and the flask was purged with
nitrogen. Oxalyl
chloride (8.40 mL, 98.0 mmol, 1.4 equiv) was added, and the reaction was
cooled to -78 C. DMSO
(11.60 mL, 0.16 mol, 2.3 equiv) was added to the addition funnel and then
added dropwise slowly at a
rate to control the vigorous gas evolution. After addition, the reaction was
stirred at -78 C for 10
min. The deacylated material was dissolved in dichloromethane (160 mL) and
added slowly to the
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reaction via addition funnel. The reaction was stirred for 10 min at -78 C.
Triethylamine (68.3 mL,
0.49 mol, 7 equiv) was then added dropwise via addition funnel. Upon
completion, the reaction was
stirred at -78 C for 10 min, warmed to room temp, and quenched with 1 M HC1.
After separation,
the organic layer was dried over Na2SO4 and concentrated in a fume hood.
Purification via flash
chromatography yielded (1R,4R,7R)-7-bromo-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptane-3,6-
dione (5) as a beige solid (13.5 g, 41.7 mmol, 60% yield).
Method C
To (1R,4R,6S,7R)-7-bromo-2-(4-methoxybenzy1)-3-oxo-2-zabicyclo[2.2.11heptan-6-
y1 acetate
(4) (96.5 g, 262.0 mmol, 1 equiv) in Me0H (680 mL) was added K2CO3 (108.6 g,
786.2 mmol, 3
equiv). The mixture was stirred at 25 C for 1 h. TLC (petroleum ether: ethyl
acetate = 1:1, Rf = 0.65)
showed that the reaction was complete. The mixture was filtered, and the
filtrate was concentrated
under reduced pressure to give a black brown solid. MTBE (500 mL) was added to
the brown solid,
and the mixture was stirred for 3 h, then filtered to give a filtrate, which
was concentrated under
reduced pressure to give the crude product. The crude compound (76.5 g, 234.5
mmol, 89.5%
yield) was obtained as an off-white solid, which was used in the next step
without further purification.
The above crude compound (76.5 g, 234.5 mmol, 1 equiv) was added to MeCN (400
mL).
IBX (65.6 g, 234.5 mmol, 1.0 equiv) was added in one portion to the above
mixture at 20-30 C for
0.5 h. The reaction mixture was stirred at 75-80 C for 2 h after which time a
TLC (petroleum ether:
ethyl acetate = 3:1, Rf = 0.23) showed the reaction was completed. The mixture
was concentrated
under reduced pressure to give the crude product, which was purified by column
chromatography on
silica gel with petroleum ether: ethyl acetate (20:1-1:1) to give (1R,4R,7R)-7-
bromo-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione (5) (48.5 g, 149.6 mmol,
63.7% yield) as an
off-white solid.
Example 3(a)
Manufacture of (1R,4R,6S,7R)-7-bromo-6-hydroxy-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptan-
3-one (25)
(1R, 4R, 6S,7R)-(+)-7-Bromo-2-( 4-methoxybenzy1)-3 -oxo-2-azabicyclo42 .2 .1] -
heptan-6-y1
acetate (4) (22.25 g, 60.42 mmol, 1 equiv) was dissolved in a solvent mixture
of methanol (120 ml)
and water (15 ml) in a 500 ml round bottom flask. Potassium carbonate powder
(12.5 g, 90.45 mmol,
1.5 equiv) was added into the golden yellow solution and the resulting cloudy
reaction mixture was
sealed with a serum cap/N2 needle and stirred overnight at room temperature.
The progress of the
reaction was monitored by lcms and by hplc. The reaction was worked up by
evaporating the volatiles
(rotary evaporator/vacuum/water bath at 40 C). Distilled water (200 ml) was
added to the dark
golden yellow oil obtained from the evaporation. The pH of the aqueous layer
was adjusted to pH= 7
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with 1M hydrochloric acid and the product extracted into dichloromethane (200
ml, then 2 x 150 m1).
The dichloromethane extracts were combined and washed with water (200 ml),
brine (250 ml), dried
with Na2SO4 powder. The dried extract was filtered and the spent drying agent
was rinsed with
dichloromethane (2 x 50 ml) and the combined extracts were concentrated
(rotary evaporator/vacuum/
water bath at 40 C) to a golden yellow oil. The golden yellow product was
split into two roughly
equal portions and each portion was purified by chromatography on the Biotage
on a 100 g silica
column, with an Et0Achexanes gradient po% (1 CV) 20-100% (7 CV) 100% (5 CV)],
fractions
collected in 25 x 150 mm tubes. The product fractions were combined and
concentrated (rotary
evaporator/vacuum/water bath at 40 C) to a golden yellow liquid which was
pumped down on the
vacuum line for several hours. The two chromatography purified samples (20 g)
were combined and
crystallized from hexanes-Et0Ac. A first crop of crystals was collected (17
g). The mother liquors
were concentrated and crystallized from hexanes-Et0Ac to provide a second crop
of crystals (2 g).
(1R,4R,6S,7R)-(+ )-7-Bromo-6-hydroxy-2-( 4-methoxybenzy1)-2-aza-bicyclo [2
.2.1 ]heptan-3-one
(25) was obtained as a white crystalline powder (total of -15 g; 75 %).
Example 3(b)
Manufacture of (1R,4R,7R)-7-Bromo-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptane-
3,6-dione (5) by Swern Oxidation
A 1-liter 3-neck round bottom flask, fitted with overhead stirrer, 50 ml
dropping funnel and
serum cap/thermocouple/N2 needle, was charged with a solution of oxalyl
chloride (3.4 g (2.3 ml),
1.25 equiv) and dichloromethane (90 ml), and the stirred acid chloride
solution was cooled in an
acetone/CO2cooling bath to -60 C. The dropping funnel was charged with a
solution of DMSO (2.6
g(2.35 ml), 1.5 equiv) in dichloromethane (18 ml) which was added into the
cold acid chloride
solution in 3 minutes. The resulting mixture was stirred for 7 minutes at -60
C and then to the
solution, (1R,4R,6S,7R)-(+)-7-bromo-6-hydroxy-2-(4-methoxybenzy1)-2-
azabicyclop.2.1]-heptan-3-
one (25) (7.2 g, 22.1 mmol, 1 equiv) was added in 10 minutes. The reaction
mixture was kept at -60
C for 25 minutes and then neat trimethylamine (21.5 ml, 7 equiv) was placed in
the dropping funnel
and added into the flask. The resulting milky white solution was kept cold for
1 hour, the reaction
checked by hplc/lcms and then the reaction was allowed to warm to room
temperature in 2.5 hours
before the work up. Saturated brine (125 ml) was added to the flask followed
by diethyl ether (125
ml) in the work up The quenched reaction was transferred into a 1-L separating
funnel. The upper
layer was saved and the aqueous lower layer was extracted with diethyl ether
(2 x 125 m1). These
ethereal extracts were combined with the original extract and this was then
washed with 1 M
hydrochloric acid (125 ml), brine (125 ml) and water (50 m1). The organic
phase was dried over
anhydrous Na2SO4powder, filtered and the filter cake washed with
dichloromethane (2 x 50 ml) and
the wash combined with the filtrate. The volatiles were removed (rotary
evaporator/vacuum/ water
bath at 40 C) to afford a yellow liquid that solidified on standing. The
solid was crystallized from
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Et0Achexanes and afforded a 1st crop (5.31 g). The mother liquors were
concentrated and
crystallized to give a 2nd crop (1.36 g). The mother liquors from the 2nd
crystallization were purified
on the Biotage with a 25g silica column eluted with Et0Ac-hexanes gradient
[20% (2 CV), 20-100%
(8 CV), 100% (2 CV)] fractions collected in 16 x 150 mm tubes. From the
chromatography 250 mg of
product was obtained. The combined yield of (1R,4R,7R)-(+)-7-bromo-2-(4-
methoxybenzy1)-
2-azabicyclo[2.2.11heptane-3,6-dione (5) was 6.8 g (94% yield).
Example 3(c)
Manufacture of (1R,4R,7R)-7-Bromo-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptane-
3,6-dione (5) by TPAP Oxidation
A 2-liter 3-neck round bottom flask fitted with overhead stirrer and serum
cap/thermocouple/N2 needle was charged with (1R,4R,6S,7R)-(+)-7-bromo-6-
hydroxy-2-(4-
methoxybenzy1)-2-azabicyclo[2.2.1]-heptan-3-one (25) (12 g, 36.8 mmol, 1
equiv), 4-morpholine-N-
oxide (12.9 g, 110 mmol, 3 equiv), and 4A molecular sieve powder in methylene
chloride ( 400 m1).
A catalytic amount of TPAP (25 mg, 0.07 mmol, 0.002 equiv) was then added. The
solution was
stirred under a nitrogen at room temperature as a green solution which
darkened over time. The
reaction was monitored daily by lc-ms. On day 2, the reaction had stalled and
the reaction was filtered
through Celite0 pad using a filter flask and vacuum source. The solution was
returned to the 2-liter 3-
neck round bottom reaction flask and fresh 4-morpholine-N-oxide (5.5 g), 4A
molecular sieve powder
(10 g) and TPAP (25 mg). The reaction had stalled late on Day 3 and the
reaction was filtered and
restarted with fresh NMO (5.5 g), 4A-sieves (10 g) and TPAP (25 mg). At the
end of Day 4 there was
¨ 7% bromoalcohol left and so the TPAP oxidation reaction was worked up. The
solids in the reaction
solution were removed by filtering with a plastic sinter funnel packed with
1/2-cm sand, 1/2-cm
anhydrous Na2SO4 powder and 1-cm depth of Celite using a Buchner flask and
vacuum source. The
filter cake was washed with dichloromethane (2 x 100 ml and 50 ml) until the
filtrate stream was
colorless. The filtrate was concentrated to a dark golden oil (rotary
evaporator/vacuum/ water bath at
40 C) which showed signs of solidifying on the glass surface. The crude
product was split into half
and each half underwent chromatography on the Biotage on silica gel (100 g)
eluted with a Et0Ac-
hexanes gradient 20% (1CV), 20-100% (7.5 CV) and 100% (4CV) collecting
fractions in 25 x 150 mm
tubes. The product fractions from the two chromatographies were combined and
concentrated (rotary
evaporator/vacuum/ water bath at 40 C) to afford a white powder.
The powder was crystallized from Et0Ac-hexanes affording a 1st crystal crop
(8.5 g). The mother
liquors were concentrated and crystallized to give a 2nd crop (1.5 g). The
combined yield of
(1R,4R,7R)-(+ )-7-bromo-2-( 4-methoxybenzy1)-2-azabicyclo[2.2.1 ]heptane-3,6-
dione (5) was 10 g
(80%).
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Example 4
Manufacture of (1R,4R,7R)-7-Bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptan-3-one (6)
Method A
(1R,4R,7 R)-7 -Bromo-2-(4-methoxybenzy1)-2-azabicyclo[2.2.11heptane-3,6-dione
(5) (1.00 g,
3.09 mmol) and 2-((difluoromethyl)sulfinyl)pyridine (20), (715.10 mg, 3.70
mmol, 1.2 equiv) were
added to a round bottom flask and purged with argon. DMF (15 mL) was added,
and the reaction was
cooled to between -55 and -65 C. KO'Bu (623.0 mg, 5.55 mmol, 1.8 equiv, 0.5 M
in DMF) was
added via syringe pump over 1 h. The temperature was maintained between -55
and -65 C. After
addition was complete, the reaction was further stirred for 30 min at -60 C.
Saturated NH4C1 (5.00
mL) was added, and the reaction was stirred for 5 min at -60 C before 6 M HC1
(5.00 mL) was
added. After 5 min of stirring at -60 C, the reaction mixture was warmed to
room temperature and
then to 65 C for 1 h. After being cooled, the reaction was diluted with
brine, extracted with ethyl
acetate (2 x 20 mL), and washed with brine (10 mL). Drying over Na2SO4 and
concentration yielded a
yellow oil, which was purified via flash chromatography to yield a white solid
(620.0 mg, 1.73 mmol,
58% yield). [a]r = -46.6 4 (c 0.80, CHC13); m.p. 85-87 C; 1H NMR (500 MHz,
CDC13) 6 7.14 (d,
J= 8.4 Hz, 1H), 6.86 (d, J= 8.5 Hz, 1H), 4.60 (d, J = 14.6 Hz, 1H), 4.19 (s,
OH), 4.14 (s, OH), 3.90 (d,
J= 14.7 Hz, 1H), 3.79 (s, 1H), 3.00 (s, OH), 2.83 (dq, J= 14.6, 3.0 Hz, 1H),
2.27 (d, J = 15.2 Hz, 1H).
See, FIG. 1. 13c NMR (126 MHz, CDC13) 6 173.0, 159.5, 153.9 (dd, J= 287.5,
283.8 Hz), 129.7,
127.5, 114.3, 87.1 (dd, J = 24.9, 23.5 Hz), 63.4, 63.3, 55.3, 50.8 (d,J= 17.1
Hz), 44.6, 24.8. See, FIG.
2. 19F NMR (376 MHz, CDC13) 6 -88.15 (dp, J= 55.1, 2.7 Hz), -88.88 (dq, J=
54.8, 2.8 Hz). See,
FIG. 3. IR (film, cm') 3013, 1785, 1683, 1551; HMRS (ESI) calc'd for
Ci5tli4BrF2NO2+Na+:
380.0074; found 380.0075.
Method B
2-(Difluoromethylsulfonyl)pyridine (20) (2.5 gm,12.94 mmol, 1.2 equiv) and
(1R,4R,7R)-( +
)-7-Bromo-2-( 4-methoxy-benzy1)-2-azabicyclo [2.2.1 lheptane-3,6-dione (5)
(3.5 gm, 10.8 mmol, 1
equiv) were combined in a round bottom flask with stir bar to which
dimethylformamide (32 ml) was
added. The resulting yellow solution was sealed with a serum cap/N2
needle/thermocouple and cooled
to -40 C. Potassium bis(trimethylsilyl)amide (KHMDS) solution (15.1 ml, 15.1
mmol, 1.4 equiv)
was added keeping the temperature between -40 C and -35 C. The reaction
solution turned a dark
orange color as the base was added. The resulting reaction mixture was stirred
for I-hour at -40 C and
then allowed to warm to room temperature gradually. The reaction was quenched
with saturated
NH4C1 solution (10 ml) , stirred for 10 minutes, 3M hydrochloric acid (40 ml)
was added, stirred for
minutes and then the flask contents were heated in a heating bath at 60 C for
1.5 hours and then
cooled to room temperature. Water (35 ml) was added and the reaction was
extracted with tert-butyl
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methyl ether (MTBE) (3 x 75 m1). The ethereal extracts were combined into one
and washed with
brine, dried with anhydrous Na2SO4 powder and filtered through a Chem-R-Us
plastic sinter funnel to
remove the spent drying agent. The filtrate was concentrated (rotary
evaporator/vacuum/ water bath at
40 C) to afford a light brown colored oil. The crude product was purified by
chromatography on the
Biotage on a 100 g silica column, an Et0Ac-hexanes gradient [15% (1 CV) 15-75%
(7.5 CV) 100%
(3 CV)] and fractions collected in 16 x 100 mm tubes. The product fractions
were combined and
concentrated (rotary evaporator/vacuum/ water bath at 40 C) to a golden
yellow liquid that was
pumped down on the vacuum line for several hours, yielding (1R,4R,7R)-(+ )-7-
bromo-6-(
difluoromethylene)-2-( 4-methoxybenzy1)-2-azabicyclo[2.2.1] heptane-3-one (6)
(2.5 g; 63 %).
Method C
To a solution of (1R,4R,7R)-7-bromo-2-(4-methoxybenzy1)-2-
azabicyclo[2.2.11heptane-3,6-
dione (5) (48.5 g, 149.6 mmol, 1 equiv) and 2-
((difluoromethyl)sulfinyl)pyridine (20), (31.8 g, 164.6
mmol, 1.1 equiv) in DMF (800 mL) was added dropwise a solution of t-BuOK (30.2
g, 269.3 mmol,
1.8 equiv) in DMF (800 mL) very slowly at -57 to -52 C. The reaction was
stirred at -55 C for 0.5 h,
then a saturated NH4C1 solution (400 mL) was slowly added dropwise to the
reaction mixture at a
temperature below -30 C. After addition, 6 N HC1 solution (360 mL) was added
to the above mixture
below -20 C. The mixture was warmed to 10 C and stirred at 65 C for 1 h.
TLC (petroleum ether:
ethyl acetate = 3:1, Rf= 0.50) showed a new spot. The mixture was added to
water (4 L) and extracted
with MTBE (2 L x 2). The organic layer was separated and washed with brine (2
L), then dried with
Na2SO4, concentrated under reduced pressure to give a residue as a red oil.
The crude product was
purified by column chromatography on silica gel with petroleum ether: ethyl
acetate (20:1-1:1) to
give (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-azabicyclo
[2.2.11heptan-3 -
one (6) (32.5 g, 90.7 mmol, 60.6% yield) as a white solid.
Example 5
Manufacture of (1R,4R,7R)-7-Bromo-6-(difluoromethylene)-2-
azabicyclo[2.2.11heptan-3-one (7)
Method A
(1R,4R,7R)-7 -B romo-6-(difluoromethylene)-2-(4-methoxybenzy1)-2-
azabicyclop.2.11heptan-
3-one (6) (140 mg, 0.39 mmol) was added to MeCN (2.0 mL) and cooled to 0 C.
Ceric ammonium
nitrate (643.5 mg, 1.17 mmol, 3 equiv) in H20 (0.75 mL) was added dropwise.
The reaction was
allowed to warm to room temperature and stirred for 1 h. After completion,
water was added, and the
solution was extracted with ethyl acetate (2 x 15 mL). The organic layers were
dried over Na2SO4 and
concentrated under reduced pressure. Flash chromatography yielded (1R,4R,7R)-7-
bromo-6-
(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7) as a white solid (75
mg, 0.315 mmol, 80%
yield). [a]iSrc= +38.5 (c 0.90, CHC13); m.p. 139-141 C; 1H NMR (500 MHz,
CDC13) 6 5.87 (s, 1H),
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4.41 (s, 1H), 4.32 (s, 1H), 2.96 (s, 1H), 2.87 (dq, J= 15.4, 3.4 Hz, 1H), 2.32
(d, J= 15.2 Hz, 1H). See,
FIG. 4. 13C NMR (126 MI-lz, CDC13) 6 174.9, 153.5 (t, J= 287.5 Hz), 88.7 (t,
J= 24.1 Hz), 60.6,
51.5, 50.3, 24.3. See, FIG. 5. 19F NMR (376 MHz, CDC13) 6 -88.60 (dq, J= 55.1,
2.8 Hz), -88.88 (dp,
J= 54.6, 2.5 Hz). See, FIG. 6. IR (film, cm-1) 3249. 1788, 1678, 1397; HMRS
(ESI+) calc'd for
C7H6BrF2NO+H+: 237.9679; found 237.9678.
Method B
An aqueous solution of Cerium (IV) ammonium nitrate (CAN) (6.4 g) 11.67 mmol,
2.94
equiv) in 20 ml distilled water was prepared 10 minutes prior to the running
the oxidative cleavage
reaction. (1R,4R,7R)-( + )-7-Bromo-6-( difl uoromethylene )-2-( 4-
methoxybenzy1)-2-aza-bicyclo[2 .2
.1 ]heptane-3-one (1.4 g, 3.91 mmol, 1 eqiv) was dissolved in acetonitrile (70
ml) and stirred with a
stir bar in a round bottom flask. The aqueous CAN solution was added to the
flask drop by drop and
the resulting mixture stirred at room temperature for hours, monitoring the
progress of the reaction by
hplc and lcms. The reaction was worked up by pouring the reaction into a 500
ml separatory funnel
and then Et0Ac (300 ml) was added, shaken and the upper layer was saved. The
lower aqueous layer
was extracted with more Et0Ac (2 x 50 m1). The ethyl acetate extracts were
combined into one and
washed with water ( 4 x 10 ml) and with saturated brine (25 ml) then dried
over anhydrous Na2SO4.
After filtering through a plastic sinter funnel to remove the spent drying
agent, the filtrate was
concentrated (rotary evaporator/vacuum/water bath at 40 C) to afford a light
brown colored oil. The
crude product was purified by chromatography on the Biotage on a 25 g silica
column, an Et0Ac-
hexanes gradient [15% (1 CV) 15-75% (7.5 CV) 100% (3 CV)] and fractions
collected in 16 x 100
mm tubes. The product containing fractions were combined and concentrated
(rotary
evaporator/vacuum/ water bath at 40 C) to a golden yellow liquid that was
pumped down on the
vacuum line for several hours yielding purified (1R,4R,7R)-(+ )-7-Bromo-6-
(difluoromethylene )-2-
azabicyclo[2.2.1 lheptane-3-one (7) (0.65 g; 70 %).
Example 6
Manufacture of (1 R,4R,7 R)-7 -Bromo-6-(difluoromethylene)-2-(tert-
butoxycarbony1)-2-
azabicyclo[2.2.11heptan-3-one (8) and Methyl (S)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-l-ene-l-carboxylate (9)
Method A
(1R,4R,7R)-7-Bromo-6-(difluoromethylene)-2-azabicyclo[2.2.11heptan-3-one (7)
(890.0 mg,
3.74 mmol) was added to dichloromethane (18.0 mL) followed by the sequential
addition of Boc20
(978.8 mg, 4.49 mmol, 1.2 equiv), DMAP (45.7 mg, 0.37 mmol, 0.1 equiv), and
Et3N (0.78 mL, 5.61
mmol, 1.5 equiv). The reaction was stirred for 1 hand then was washed with 1 M
HC1 (10 mL), dried
over Na2SO4, and concentrated. The resulting oil containing (1R,4R,7R)-7-Bromo-
6-
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(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-3-one
(8) was dissolved in
methanol (18.0 mL), then K2CO3 (1.55 g, 11.21 mmol, 3.0 equiv) was added, and
the reaction was
stirred for 6 h. After completion, as indicated by LC/MS (methanolysis of the
lactam occurs in the
first 10 min), the reaction was diluted with brine and extracted with ethyl
acetate (3 x 200 mL). Upon
drying over Na2SO4, concentrating, and purification by flash chromatography,
methyl (S)-3-((tert-
butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-l-ene-l-carboxylate (9)
was obtained as a
white solid (570 mg, 1.97 mmol, 52% yield). [a]r= +104.8 (c 0.50, CHC13); m.p.
95-97 C; 1H
NMR (500 MHz, CDC13) 6 6.58 (s, 1H), 5.50 (d, J = 9.1 Hz, 1H), 4.63 (d, J= 8.7
Hz, 1H), 3.75 (s,
3H), 3.33 (d, J= 20.4 Hz, 1H), 3.21 (dd, J= 20.3, 2.7 Hz, 1H), 1.42 (s, 9H).
See, FIG. 7. 11C NMR
(126 MHz, CDC13) 6 164.3, 154.7, 154.6, 152.4 (t, J = 288.5 Hz), 150.1, 140.6,
135.5, 88.9 (dd, J =
21.8, 20.2 Hz), 80.1, 55.3, 51.9, 31.1, 28.3. See, FIG. 8. 19F NMR (376 MHz,
CDC13) 6 -84.49 (d, J =
43.6 Hz), -85.91 (d, J = 43.4 Hz). See, FIG. 9. IR (film, cm') 3347, 2987,
1773, 1681; HMRS (ESI )
calc'd for Ci3Ht7F2N04+Na+: 312.1023; found 312.1018.
Method B
To a solution of (1R,4R,7R)-7-bromo-6-(difluoromethylene)-2-
azabicyclo[2.2.11heptan-3-one
(7) (14.4 g, 60.5 mmol, 1 equiv) in DCM (150 mL) were added DMAP (739.0 mg,
6.05 mmol, 0.1
equiv) and TEA (9.18 g, 90.7 mmol, 12.6 mL, 1.5 equiv). Then (Boc)20 (15.8 g,
72.6 mmol, 16.6 mL,
1.2 equiv) was added slowly. After the addition, the mixture was stirred at 25
C for 1 h. TLC
(petroleum ether: ethyl acetate = 1:1, Rf = 0.20) showed that the reaction was
completed. The mixture
was adjusted to pH = 3-4 with 1N HC1 solution, then the organic layer was
separated and washed with
brine (100 mL), dried with Na2SO4, and concentrated under reduced pressure to
give (1R,4R,7R)-7-
bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-azabicyclo[2.2.11heptan-
3-one (8) (7.50 g,
crude) as a black brown solid, which was used in the next step without further
purification.
(1R,4R,7 R)-7 -Bromo-6-(difluoromethylene)-2-(tert-butoxycarbony1)-2-
azabicyclo[2.2.11heptan-3-one
(8) (7.50 g, 22.1 mmol, 1 equiv) was dissolved in Me0H (55 mL), then CH3ONa
(1.44 g, 26.6 mmol,
1.2 equiv) was added. The mixture was stirred at 0 C for 1 h. TLC (petroleum
ether: ethyl acetate =
1:1, Rf = 0.70) showed that the reaction was completed. Water (100 mL) was
added to the reaction
mixture, and the mixture was extracted with MTBE (200 mL x 2), washed with
brine (150 mL), dried
with Na2SO4, filtered, and concentrated under reduced pressure to give a
residue as a red solid
(crude). The crude product was purified by column chromatography on silica gel
with petroleum
ether: ethyl acetate (20:1-1:1) to give methyl (5)-3-((tert-
butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylate (9) (5.20 g, 17.9 mmol, 81.0%
yield) as a white
solid.
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Example 7
Manufacture of tert-Butyl (1R,4R,7R)-(+)-7-bromo-6-(difluoromethylene)-
3-oxo-2-azabicyclo[2.2.11heptan-2-carboxylate (8)
Dichloromethane (30 ml) used in this reaction was deoxygenated with a gas
dispersion tube
and nitrogen just prior to use in the reaction. (1R,4R,7R)-(+)-7-Bromo-6-
(difluoromethylene)-2-
azabicyclo[2.2.1 1-heptan-3-one (7) (1.4 g, 5.88 mmol, 1 equiv) and di-tert-
butyl dicarbonate (2.56 g,
11.73 mmol, 2 equiv) were dissolved in dichloromethane (30 ml) in a 100 ml
round bottom flask. 4-
(Dimethylamino)-pyridine (DMAP) (0.3 g, 2.46 mmol, 7.3 equiv) was dissolved in
triethylamine (6
ml, 43 mmol, 0.4 eqiv) and then the amines were added to the reaction drop by
drop under a nitrogen
atmosphere blanket. The resulting reaction mixture in the flask was sealed
with a serum cap/N2 needle
and the contents was stirred for 1 hour at room temperature. A small aliquot
was removed and the
reaction progress was checked by hplc and lcms. In the work up, the volatiles
were removed (rotary
evaporator/vacuum/water bath at ambient temperature) to afford a dark golden
yellow liquid. The
liquid was taken up in dichloromethane (50 ml) and partitioned with 1 M
hydrochloric acid (25 ml),
washed with brine (50 m1). The extract was dried over anhydrous Na2SO4 powder,
filtered and the
filtrate concentrated (rotary evaporator/vacuum/water bath at ambient
temperature) to afford a brown
oil. The crude product was purified by chromatography on the Biotage on a 25 g
silica column, an
Et0Ac-hexanes gradient [15% (1 CV) 15-75% (7.5 CV) 100% (3 CV)] and fractions
collected in 16 x
100 mm tubes. The product containing fractions were combined and concentrated
(rotary
evaporator/vacuum/ water bath at 40 C) to a golden yellow liquid that was
pumped down on the
vacuum line for several hours yielding purified tert-butyl (1R,4R,7 R)-(+ )-7-
Bromo-6-
(difluoromethylene )-3-oxo-2-azabicyclo[2.2.1 lheptane-2-carboxylate (1.65 g;
82%).
Example 8
Manufacture of (S)-3-((tert-butoxycarbonyl)amino)-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylic acid (19)
The solvent methanol-water (3:1 v/v) used in this reaction was deoxygenated
with a gas
dispersion tube and nitrogen just prior to use in the reaction. tert-Butyl
(1R, 4R,7 R)-( + )-7-bromo-6-
(difluoromethylene )-3-oxo-2-azabicyclo[2.2.1 [-heptane-2-carboxylate (8) (1.6
g, 4.73 mmol, 1
equiv) was dissolved in the aqueous methanol (35 ml) and stirred with a stir
bar under a nitrogen
atmosphere, and potassium carbonate powder (1.96 g, 14.18 mmol, 3 equiv) was
added into the
solution in a single portion. The reaction flask was sealed with a serum cap
and nitrogen needle and
the mixture inside stirred for 12 hours. Dichloromethane (50 ml) was added and
then the biphasic
crude reaction mixture was acidified with 1 M hydrochloric acid to destroy the
excess potassium
carbonate and bring the pH to 7 (pH papers). The phases were split and then
the volatiles were
removed from the dichloromethane layer. The crude product was purified by
chromatography on the
49
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Biotage on a 25 g silica column, a methanol-CH2Ch gradient [1 % (1 CV) 1-20%
(7.5 CV) 20% (3
CV)] and fractions collected in 16 x 100 mm tubes. The product containing
fractions were combined
and concentrated (rotary evaporator/vacuum/ water bath at 40 C) to a golden
yellow solid that was
pumped down on the vacuum line for several hours to yield purified (S)-3-
((tert-
butoxycarbonyl)amino )-4-( difluoromethylene )cyclopent-l-ene-l-carboxylic
acid (19) (0. 733 g; 56
0/0).
Example 9
Manufacture of (S)-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic
acid hydrochloride
(1)
Method A
Methyl (S)-3-((tert-butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-
1-
carboxylate (9) (570.0 mg, 1.97 mmol) was dissolved in dioxane (1.00 mL), and
6 M HC1 (9 mL) was
added. After heating at 80 C for 2 h, the reaction was concentrated to yield
(S)-3-Amino-4-
(difluoromethylene)cyclopent-1-ene-1-carboxylic acid hydrochloride (1) as a
light brown powder
(403.0 mg, 1.90 mmol, 97% yield). Crystallization from ethanol/diethyl ether
increased purity to
>99%. [c]rc= +67.2 (e 0.90, H20); m.p. 207 C (decomp.); 1H NMR (500 MHz, D20)
6 6.59 (s,
1H), 4.70 (s, 13H), 3.39 (d, J= 20.5 Hz, 1H), 3.33 (d, J= 20.8 Hz, 1H). See,
FIG. 10. 13C NMR (126
MHz, D20) 6 167.2, 153.0 (dd, J = 290.1, 288.4 Hz), 141.8, 134.3, 86.1 (dd, J
= 26.6, 21.2 Hz), 54.8
(d, J= 5.7 Hz), 31.1. See, FIG. 11. "F NMR (470 MHz, D20) 6 -83.1 (dq, J=
40.8, 2.8 Hz), -83.5
(dq, J= 40.4, 2.1 Hz). See, FIG. 12. IR (film, cm') 3348, 3075, 2981, 2883,
2829, 2600, 2434, 1771,
1686; HMRS (ES!-) calc'd for C7H7F2NO2-H: 174.0372; found 174.0369.
Method B
(S)-3-((tert-Butoxycarbonyl)amino)-4-(difluoromethylene)cyclopent-1-ene-1-
carboxylate (9)
(5.20 g, 17.9 mmol, 1 equiv) was added to 4 M HC1(g)/Et0Ac (100 mL), and the
mixture was stirred
at 20-30 C for 2 h. TLC (petroleum ether: ethyl acetate = 5:1, Rf= 0)
indicated the starting
material was consumed completely. The mixture was concentrated under reduced
pressure to give
methyl (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylate
hydrochloride (3.00 g,
crude, HC1 salt) as a yellow solid, which was used in the next step without
further purification. The
above compound (3.00 g, 13.3 mmol, 1.0 equiv) was added to Me0H (15 mL), and a
solution of
LiORH20 (1.39 g, 33.2 mmol, 2.5 equiv) in H20 (10 mL) was added to the above
mixture slowly at
20-30 C. The mixture was stirred at 20-30 C for 3 h. TLC (dichloromethane:
methanol = 5:1 ,Rf=
0.40) indicated that the starting material was consumed completely. The
mixture was adjusted to pH
= 2-3 with 6 M HC1 solution and concentrated to give the crude product, which
was purified by
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reversed-phase HPLC (Agela C18 330 g; mobile phase: [water (0.1%HC1)-Me0H1;
B%: 10%-20%,
20 min, 70 mLimin) to give (S)-3-amino-4-(difluoromethylene)cyclopent- 1-ene-
1-carboxylic acid
hydrochloride (1) (1.80 g, 8.46 mmol, 63.7% yield, 99.5% purity) as a white
solid.
Example 10
Manufacture of (S)-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-
carboxylic acid hydrochloride (1)
6N Hydrochloric acid was prepared from mixing conc. hydrochloric acid (10 ml)
and water
(10 m1). THF used in this reaction was deoxygenated with a gas dispersion tube
and nitrogen just
prior to use in the reaction. (S)-3-((tert-butoxycarbonyl)amino )-4-
(difluoromethylene )-cyclopent-l-
ene-l-carboxylic acid (19) was dissolved in THF (2 ml) and stirred in a 10 ml
round bottom flask with
a stir bar. 6M hydrochloric acid (2 ml) was added. The reaction mixture was
stirred at room
temperature for 2 hours and then the THF and water was removed with an active
nitrogen gas flow
overnight. The pink colored solid residue was pumped down on a vacuum line for
several hours to
remove any residual solvent yielding (S)-3-amino-4-(difluoromethylene )-1-
cyclopentene-l-carboxylic
acid hydrochloride salt (1) as a pink solid (-100 mg; 86%).
It should be understood that the examples and embodiments provided herein are
exemplary
examples and embodiments. Those skilled in the art will envision various
modifications of the
examples and embodiments that are consistent with the scope of the disclosure
herein. Such
modifications are intended to be encompassed by the claims.
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