Note: Descriptions are shown in the official language in which they were submitted.
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3-AMINO-2-HYDROXYALKANOIC ACIDS AND THEIR PRODRUGS
Technical Field
The present invention relates to novel compounds having methionine
aminopeptidase-
2 inhibitory (MetAP2) activity useful for treating conditions which arise from
or are
exacerbated by angiogenesis, pharmaceutical compositions comprising the
compounds,
l0 methods of treatment using the compounds, methods of inhibiting
angiogenesis, and methods
of treating cancer.
Packground of the Invention
Angiogenesis is the fundamental process by which new blood vessels are formed
and
15 is essential to a variety of normal body activities (such as reproduction,
development, and
wound repair). Although the process is not completely understood, it is
believed to involve a
complex interplay of molecules which both stimulate and inhibit the growth of
endothelial
cells, the primary cells of the capillary blood vessels. Under normal
conditions these
molecules appear to maintain the microvasculature in a quiescent state (i.e.,
one of no
2o capillary growth) for prolonged periods that may last for weeks, or in some
cases, decades.
However, when necessary, such as during wound repair, these same cells can
undergo rapid
proliferation and turnover within as little as five days.
Although angiogenesis is a highly regulated process under normal conditions,
many
diseases (characterized as "angiogenic diseases") are driven by persistent
unregulated
25 angiogenesis. Otherwise stated, unregulated angiogenesis may either cause a
particular
disease directly or exacerbate an existing pathological condition. Thus, there
is a continuing
need for compounds which demonstrate antiangiogenic activity.
Summary of the Invention
3o In its principle embodiment, the present invention provides a compound of
formula (17
R1 O
H2N OR2
OH
(
or a therapeutically acceptable salt thereof, wherein
Rl is selected from the group consisting of alkyl, alkylsulfanylalkyl, aryl,
arylalkyl,
35 cycloalkyl, (cycloalkyl)alkyl, (heterocycle}alkyl, and hydroxyalkyl; and
-1-
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R2 is selected from the group consisting of hydrogen, alkenyl, alkyl,
alkylcarbonyloxyalkyl, alkylcarbonylalkyl, aryl, arylalkyl, cycloalkyl,
(cycloalkyl)alkyl,
heterocycle, and (heterocycle)alkyl.
In another embodiment, the present invention provides a phal-maceutical
composition
comprising a compound of formula (I), or a therapeutically acceptable salt
thereof, in
combination with a therapeutically acceptable carrier.
In another embodiment, the present invention provides a method of inhibiting
angiogenesis in a mammal in recognized need of such treatment comprising
administering to
the mammal a pharmaceutically acceptable amount of a compound of formula (1?.
i0 In another embodiment, the present invention provides a method for treating
cancer in
a patient in recognized need of such treatment comprising administering to the
patient a
therapeutically acceptable amount of a compound of formula (I), or a
therapeutically
acceptable salt thereof.
15 Detailed Description of the Invention
As used herein, the singular forms "a", "an", and "the" include plural
reference unless
the context clearly dictates otherwise.
As used in the present specification the following terms have the meanings
indicated:
The term "alkenyl," as used herein, refers to a monovalent straight or
branched chain
20 group of one to ten carbon atoms containing at least one carbon-carbon
double bond.
The term "alkoxy," as used herein, refers to an alkyl group attached to the
parent
molecular moiety through an oxygen atom.
The term "alkyl," as used herein, refers to a group derived from a straight or
branched
chain saturated hydrocarbon of one to ten atoms.
25 The term "alkylcarbonyl," as used herein, refers to an alkyl group attached
to the
parent molecular moiety through a carbonyl group.
The term "alkylcarbonyloxy," as used herein, refers to an alkyIcarbonyl group
attached
to the parent molecular moiety through an oxygen atom.
The term "alkylcarbonyloxyaIkyl," as used herein, refers to an
alkylcarbonyloxy group
30 attached to the parent molecular moiety through an alkyl group.
The teim "alkylsulfanyl," as used herein, refers to an alkyl group attached to
the parent
molecular moiety through a sulfur atom.
The term "alkylsulfanylalkyl," as used herein, refers to an alkylsulfanyl
group attached
to the parent molecular moiety through an alkyl group.
35 The term "amino," as used herein, refers to -NRaRb, wherein Ra and Rb are
independently selected from the group consisting of hydrogen, alkyl,
alkylcarbonyl,
cycIoalkyl, (cycioaIkyI)alkyI, and unsubstituted aryl.
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The term "aryl," as used herein, refers to a phenyl group, or a bicyclic or
tricyclic
fused ring system wherein one or more of the fused rings is a phenyl group.
Bicyclic fused
ring systems are exemplified by a phenyl group fused to a monocyclic
cycloalkenyl group, as
defined herein, a monocyclic cycloalkyl group, as defined herein, or another
phenyl group.
Tricyclic fused ring systems are exemplified by a bicyclic fused ring system
fused to a
monocyclic cycloalkenyl group, as defined herein, a monocyclic cycloalkyl
group, as defined
herein, or another phenyl group. Representative examples of aryl include, but
are not limited
to, anthracenyl, azulenyl, fluorenyl, indanyl, indenyl, naphthyl, phenyl, and
tetrahydronaphthyl. The aryl groups of the present invention can be optionally
substituted
to with one, two, three, four, or five substituents independently selected
from the group
consisting of alkoxy, alkyl, alkylcarbonyl, amino, carboxy, cyano, halo,
haloalkoxy, haloalkyl,
vitro, and oxo.
The term "arylalleyl," as used herein, refers to an aryl group attached to the
parent
molecular moiety through an alkyl group. The alkyl part of the arylalkyl can
be optionally
substituted with an amino group.
The term "carbonyl," as used herein, refers to -C(O)-.
The term "carboxy," as used herein, refers to -C02H.
The term "cyano," as used herein, refers to -CN.
The term "cycloalkenyl," as used herein, refers to a non-aromatic cyclic or
bicyclic
2o ring system having three to ten carbon atoms and one to three rings,
wherein each five-
membered ring has one double bond, each six-membered ring has one or two
double bonds,
each seven- and eight-membered ring has one to three double bonds, and each
nine-to ten-
membered ring has one to four double bonds. Examples of cycloalkenyl groups
include, but
are not limited to, cyclohexenyl, octahydronaphthalenyl, and norbornylenyl.
The term "cycloalkyl," as used herein, refers to a saturated monocyclic,
bicyclic, or
tricyclic hydrocarbon ring system having three to twelve carbon atoms.
Examples of
cycloalkyl groups include, but are not limited to, cyclopropyl, cyclopentyl,
bicyclo[3.1.1]heptyl, and adamantyl.
The term "(cycloalkyI)alkyl," as used herein, refers to a cycloalkyl group
attached to
3o the parent molecular moiety through an alkyl group.
The term "halo," or "halogen," as used herein, refers to F, Cl, Br, or I.
The term "haloalkoxy," as used herein, refers to a haloalkyl group attached to
the
parent molecular moiety through an oxygen atom.
The term "haloalkyl," as used herein, refers to an alkyl group substituted by
one, two,
three, or four halogen atoms.
The term "heterocycle," as used herein, refers to a five-, six-, or seven-
membered ring
containing one, two, or three heteroatoms independently selected from the
group consisting of
-3-
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nitrogen, oxygen, and sulfur. The five-membered ring has zero to two double
bonds and the
six- and seven-membered rings have zero to three double bonds. The term
"heterocycle" also
includes bicyclic groups in which the heterocycle ring is fused to a phenyl
group, a
monocyclic cycIoalkenyl group, as defined herein, a monocyclic cycloalkyl
group, as defined
herein, or another monocyclic heterocycle group, as defined herein; and
tricyclic groups in
which a bicyclic system is fused to a phenyl group, a monocyclic cycloalkenyl
group, as
defined herein, a monocyclic cycloalkyl group, as defined herein, or another
monocyclic
heterocycle group. The heterocycle groups of the present invention can be
attached to the
parent molecular moiety through a carbon atom or a nitrogen atom in the group.
Examples of
heterocycles include, but are not limited to, benzothienyl, furyl, imidazolyl,
indolinyl, indolyl,
isothiazolyl, isoxazolyl, morpholinyl, oxazolyl, piperazinyl, piperidinyl,
pyrazolyl, pyridinyl,
pyrrolidinyl, pyrrolopyridinyl, pyrrolyl, thiazolyl, thienyl,
and.thiomorpholinyl. The
heterocycle groups of the present invention can be optionally substituted with
one, two, three,
four, or five substituents independently selected from the group consisting of
alkoxy, alkyl,
alkylcarbonyl, amino, carboxy, cyano, halo, haloalkoxy, haloalkyl, nitro, and
oxo.
The term "(heterocycle)alkyl," as used herein, refers to a heterocycle group
attached to
the parent molecular moiety through an alkyl group.
The term "nitro," as used herein, refers to -h.T02.
The term "oxo," as used herein, refers to =O.
2o The compounds of the present invention can exist as therapeutically
acceptable salts.
The term "therapeutically acceptable salt," as used herein, represents salts
or zwitterionic
forms of the compounds of the present invention which are water or oil-soluble
or dispersible,
which are suitable for treatment of diseases without undue toxicity,
irritation, and allergic
response; which are commensurate with a reasonable benefit/risk ratio, and
which are
effective for their intended use. The salts can be prepared during the final
isolation and
purification of the compounds or separately by reacting an amino group with a
suitable acid.
Representative acid addition salts include acetate, adipate, alginate,
citrate, aspartate,
benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate,
digluconate,
glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, fumarate,
hydrochloride,
3o hydrobromide, hydroiodide, 2-hydroxyethansulfonate, lactate, maleate,
mesitylenesulfonate,
methanesulfonate, naphthylenesulfonate, nicotinate, 2-naphthalenesulfonate,
oxalate,
pamoate, pectinate, persulfate, 3-phenyIproprionate, picrate, pivalate,
propionate, succinate,
tartrate, trichloroacetate, trifluoroacetate, phosphate, glutamate,
bicarbonate, para-
toluenesulfonate, and undecanoate. Also, ammo groups in the compounds of the
present
invention can be quaternized with methyl, ethyl, propyl, and butyl chlorides,
bromides, and
iodides; dimethyl, diethyl, dibutyl, and diamyl sulfates; decyl, Iauryl,
myristyl, and steryl
chlorides, bromides, and iodides; and benzyl and phenethyl bromides. Examples
of acids
-4-
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which can be employed to form therapeutically acceptable addition salts
include inorganic
acids such as hydrochloric, hydrobromic, sulfuric, and phosphoric, and organic
acids such as
oxalic, malefic, succinic, and citric.
Basic addition salts can be prepared during the final isolation and
purification of the
compounds by reacting a carboxy group with a suitable base such as the
hydroxide,
carbonate, or bicarbonate of a metal cation or with ammonia or an organic
primary,
secondary, or tertiary amine. The cations of therapeutically acceptable salts
include lithium,
sodium, potassium, calcium, magnesium, and aluminum, as well as nontoxic
quaternary
amine cations such as ammonium, tetramethylammonium, tetraethylammonium,
methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine,
ethylamine,
tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-
methylmorpholine,
dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine, 1-
ephenamine,
and N,N'-dibenzylethylenedianune. Other representative organic amines useful
for the
formation of base addition salts include ethylenediamine, ethanolamine,
diethanolamine,
piperidine, and piperazine.
The present compounds can also exist as therapeutically acceptable prodrugs.
The
term "therapeutically acceptable prodrug," refers to those prodrugs or
zwitterions which are
suitable for use in contact with the tissues of patients without undue
toxicity, irritation, and
allergic response, are commensurate with a reasonable benefit/risk ratio, and
are effective for
their intended use. The term "prodrug," refers to compounds which are rapidly
transformed
in vivo to parent compounds of formula (I) for example, by hydrolysis in
blood. Prodrugs of
the present invention are compounds of formula (I] where R2 is other than
hydrogen (i.e.,
carboxylic esters). Representative prodrugs include, but are not limited to,
methyl (3R)-3-
amino-2-hydroxy-5-(methylsulfanyl)pentanoate; methyl (2RS,3R)-3-amino-4-
cyclohexyl-2-
hydroxybutanoate; (1S,2R)-2-amino-2,3-dihydro-1H inden-1-yl (2RS,3R)-3-amino-4-
-
cyclohexyl-2-hydroxybutanoate; benzyl (2S,3R)-3-amino-3-cyclopentyl-2-
hydroxypropanoate; benzyl (2S,3R)-3-amino-3-cycloheptyl-2-hydroxypropanoate;
(2S)-2-
(methylamino)-2-(1-naphthyl)ethyl (2S,3R)-3-amino-5-(ethylsulfanyl)-2-
hydroxypentanoate;
(2R)-2-(methylanuno)-2-(1-naphthyl)ethyl (2S,3R)-3-amino-5-(ethylsulfanyI)-2-
3o hydroxypentanoate; 2-(methylamino)-2-(1-naphthyl)propyl (3R)-3-amino-5-
(ethylsulfanyl)-2-
hydroxypentanoate; (2R)-2-(methylamino)-2-(1-naphthyl)ethyl (2S,3R)-3-amino-4-
cyclohexyl-2-hydroxybutanoate; (2R)-2-(methylamino)-2-(1-naphthyl)ethyl
(2S,3R)-3-amino-
2-hydroxy-5-phenylpentanoate; methyl (2RS,3R)-3-amino-5-(ethylsulfanyl)-2-
hydroxypentanoate; benzyl (2RS,3R)-3-amino-5-(ethylsulfanyl)-2-
hydroxypentanoate; butyl
(2RS,3R)-3-amino-5-(ethylsulfanyl)-2-hydroxypentanoate; isopropyl (2S,3R)-3-
amino-2-
hydroxy-5-(isopropylsulfanyl)pentanoate; isopropyl (2RS,3R)-3-amino-5-
(ethylsulfanyl)-2-
hydroxypentanoate; [(2,2-dimethylpropanoyl)oxy]methyl (2RS,3R)-3-amino-5-
-5-
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(ethylsulfanyl)-2-hydroxypentanoate; sec-butyl (2RS,3R)-3-amino-5-
(ethylsulfanyl)-2-
hydroxypentanoate; (1,3-dioxo-1,3-dihydro-2H isoindol-2-yl)methyl (2RS,3R)-3-
amino-5-
(ethylsulfanyl)-2-hydroxypentanoate; sec-butyl (2RS,3R)-3-amino-2-hydroxy-5-
(isopropylsulfanyl)pentanoate; [(2,2-dimethyIpropanoyI)oxy]methyl (2S,3R)-3-
amino-2-
hydroxy-5-(isopropylsulfanyl)pentanoate; (I,3-dioxo-I,3-dihydro-ZH isoindol-
2~yI)methyl
(2S,3R)-3-amino-2-hydroxy-5-(isopropylsulfanyl)pentanoate; and 3-oxo-I,3-
dihydro-2-
benzofuran-I-yl (2S,3R)-3-amino-2-hydroxy-5-(isopropylsulfanyI)pentanoate.
Asymmetric centers exist in the compounds of the present invention. These
centers
are designated by the symbols "R" or "S," depending on the configuration of
substituents
around the chiral carbon atom. It should be understood that the invention
encompasses all
stereochemical isomeric forms, or mixtures thereof, which possess the ability
to inhibit
angiogenesis. Individual stereoisomers of compounds can be prepared
synthetically from
commercially available starting materials which contain chiral centers or by
preparation of
mixtures of enantiomeric products followed by separation such as conversion to
a mixture of
diastereomers followed by separation or recrystallization, chromatographic
techniques, or
direct separation of enantiomers on chiral chromatographic columns. Starting
compounds of
particular stereochemistry are either commercially available or can be made
and resolved by
techniques known in the art.
In accordance with methods of treatment and pharmaceutical compositions of the
invention, the compounds can be administered alone or in combination with
other anticancer
agents. When using the compounds, the specific therapeutically effective dose
level for any
particular patient will depend upon factors such as the disorder being treated
and the severity
of the disorder; the activity of the particular compound used; the specific
composition
employed; the age, body weight, general health, sex, and diet of the patient;
the time of
administration; the route of administration; the rate of excretion of the
compound employed;
the duration of treatment; and drugs used in combination with or coincidently
with the
compound used. The compounds can be administered orally, parenterally,
osmotically (nasal
sprays), rectally, vaginally, or topically in unit dosage formulations
containing carriers,
adjuvants, diluents, vehicles, or combinations thereof. The term "parenteral"
includes
infusion as well as subcutaneous, intravenous, intramuscular, and intrasternal
injection.
Parenterally administered aqueous or oleaginous suspensions of the compounds
can
be formulated with dispersing, wetting, or suspending agents. The injectable
preparation can
also be an injectable solution or suspension in a diluent or solvent. Among
the acceptable
diluents or solvents employed are water, saline, Ringer's solution, buffers,
monoglycerides,
diglycerides, fatty acids such as oleic acid, and fixed oils such as
monoglycerides or
diglycerides.
-6-
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The antiangiogenic effect of parenterally administered compounds can be
prolonged
by slowing their absorption. One way to slow the absorption of a particular
compound is
administering injectable depot forms composing suspensions of crystalline,
amoyhous, or
otherwise water-insoluble forms of the compound. The rate of absorption of the
compound is
dependent on its rate of dissolution which is, in turn, dependent on its
physical state. Another
way to slow absorption of a particular compound is administering injectable
depot forms
comprising the compound as an oleaginous solution or suspension. Yet another
way to slow
absorption of a particular compound is administering injectable depot forms
comprising
microcapsule matrices of the compound trapped within liposomes,
rnicroemulsions, or
biodegradable polymers such as polylactide-polyglycolide, polyorthoesters or
polyanhydrides.
Depending on the ratio of drug to polymer and the composition of the polymer,
the rate of
drug release can be controlled.
Transdermal patches can also provide controlled delivery of the compounds. The
rate
of absorption can be slowed by using rate controlling membranes or by trapping
the
compound within a polymer matrix or gel. Conversely, absorption enhancers can
be used to
increase absorption.
Solid dosage forms for oral administration include capsules, tablets, pills,
powders,
and granules. In these solid dosage forms, the active compound can optionally
comprise
diluents such as sucrose, lactose, starch, talc, silicic acid, aluminum
hydroxide, calcium
silicates, polyamide powder, tableting lubricants, and tableting aids such as
magnesium
stearate or microcrystalline cellulose. Capsules, tablets, and pills can also
comprise buffering
agents, and tablets and pills can be prepared with enteric coatings or other
release-controlling
coatings. Powders and sprays can also contain excipients such as talc, silicic
acid, aluminum
hydroxide, calcium silicate, polyamide powder, or mixtures thereof. Sprays can
additionally
contain customary propellants such as chlorofluorohydrocarbons or substitutes
therefore.
Liquid dosage forms for oral administration include emulsions, microemulsions,
solutions, suspensions, syrups, and elixirs comprising inert diluents such as
water. These
compositions can also comprise adjuvants such as wetting, emulsifying,
suspending,
sweetening, flavoring, and perfuming agents.
Topical dosage forms include ointments, pastes, creams, lotions, gels,
powders,
solutions, sprays, inhalants, and transdermal patches. The compound is mixed
under sterile
conditions with a carrier and any needed preservatives or buffers. These
dosage forms can
also include excipients such as animal and vegetable fats, oils, waxes,
paraffins, starch,
tragacanth, cellulose derivatives, polyethylene glycols, silicones,
bentonites, silicic acid, talc
and zinc oxide, or mixtures thereof. Suppositories for rectal or vaginal
administration can be
prepared by mixing the compounds with a suitable non-irritating excipient such
as cocoa
butter or polyethylene glycol, each of which is solid at ordinary temperature
but fluid in the
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rectum or vagina. Ophthalmic formulations comprising eye drops, eye ointments,
powders,
and solutions are also contemplated as being~within the scope of this
invention.
The total daily dose of th.e compounds administered to a host in single or
divided
doses can be in amounts from about 0.1 to about 200 mg/kg body weight or
preferably from
about 0.25 to about 100 mg/kg body weight. Single dose compositions can
contain these
amounts or submultiples thereof to make up the daily dose.
Determination of Biol ~ical Activity
Proc. Natl. Acad. Sci. USA 94: 6099-6103 (1997) and Chemistry and Biolo~y,
4(6):
461-471 (1997) report that both AGM-1470 and ovalicin, a sequiterpene isolated
from the
fungus Pseudorotiuaax ocalis have been found to bind to a common bifunctional
protein, type
2-methionine aminopeptidase (MetAP-2) and conclude that MetAP2 plays a
critical role in
the proliferation of endothelial cells and may serve as a promising target for
the development
of new anti-angiogenic drugs.
Assays for the inhibition of catalytic activity of MetAP2 were performed in 96-
well
microtiter plates. Compounds to be tested (compounds of formula (>] where R2
is hydrogen)
were dissolved in dimethyl sulfoxide at 10 mM and diluted ten-fold in assay
buffer (50 mM
HEPES, pH 7.4, 100 mM NaCI). Ten microliters of solution of each compound to
be tested
for inhibition were introduced into each cell of the plate. hero inhibition of
enzyme activity
2o was taken to be the result obtained in cells in which 10 mL of assay buffer
was placed. A
mixture totaling 90 mL per well and made up of 84 rnL of assay buffer
containing 100 mM
MnCl2, 1 rnL of L-amino acid oxidase (Sigma Catalog No. A-9378, ~11 mg/mL), 1
mL of
horseradish peroxidase (Sigma Catalog No. P-8451, dissolved in assay buffer at
a
concentration of 10 mg/mL), 1 mL of the tripeptide Met-Ala-Ser (Sachem)
dissolved in assay
buffer at concentration of 50 mM, 1 mL of oYtho-dianisidine (Sigma Catalog No.
D-1954,
freshly made solution in water at a concentration of 10 mg/mL), and MetAP2 at
a final
concentration of 1.5 mg/mL was rapidly mixed and added to each cell containing
test or
control compound. The absorbence at 450 nanometers was measured every 20
seconds over a
period of twenty minutes using an automatic plate reader (Molecular Devices,
CA, USA).
3o The Vmax in mOD/min, calculated for each well, was used to represent MetAP2
activity.
The ICSO for each inhibitor was obtained by plotting the remaining activity
versus inhibitor
concentrations. Representative compounds of the present invention had ICSO's
between about
0.030 ~M and about 1.80 ~,M. Preferred compounds of the present invention had
IC5o's
between about 0.030 and about 0.05 ~,M.
As tlxe literature has established a casual link between inhibition of MetAP2
and the
resultant inhibition of endothelial cell proliferation and angiogenesis, it
can be inferred that
the compounds of the invention, including, but not limited to those specified
in the examples,
_g_
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possess antiangiogenic activity. As angiogenesis inhibitors, such compounds
are useful in the
treatment of both primary and metastatic solid tumors, including carcinomas of
breast, colon,
rectum, lung, oropharynx, hypopharynx, esophagus, stomach, pancreas, liver,
gallbladder and
bile ducts, small intestine, urinary tract (including kidney, bladder, and
urothelium), female
genital tract (including cervix, uterus, and ovaries as well as
choriocarcinoma and gestational
trophoblastic disease), male genital tract (including prostate, seminal
vesicles, testes, and
germ cell tumors), endocrine glands (including the thyroid, adrenal, and
pituitary glands), and
skin, as well as hemangiomas, melanomas, sarcomas (including those arising
from bone and
soft tissues as well as Kaposi's sarcoma) and tumors of the brain, 'nerves,
eyes, and meninges
l0 (including astrocytomas, gliomas, glioblastomas, retinoblastomas, neuromas,
neuroblastomas,
Schwannomas, and meningiomas). Such compounds may also be useful in treating
solid
tumors arising from hematopoietic malignancies such as leukemias (i.e.,
chloromas,
plasmacytomas and the plaques and tumors of mycosis fungosides and cutaneous T-
cell
lymphoma/leukemia) as well as in the treatment of lymphomas (both Hodgkin's
and non-
Hodgkin's lymphomas). In addition, these compounds may be useful in the
prevention of
metastases from the tumors described above either when used alone or in
combination with
radiotherapy andlor other chemotherapeutic agents. The compounds of the
invention can also
be useful in the treatment of the aforementioned conditions by mechanisms
other than the
inhibition of angiogenesis.
Further uses include the treatment and prophylaxis of autoimmune diseases such
as
rheumatoid, immune and degenerative arthritis; various ocular diseases such as
diabetic
retinopathy, retinopathy of prematurity, corneal graft rejection, retrolental
fibroplasia,
neovascular glaucoma, rubeosis, retinal neovascularization due to
macular~degeneration,
hypoxia, angiogenesis in the eye associated with infection or surgical
intervention, and other
abnormal neovascularization conditions of the eye; skin diseases such as
psoriasis; blood
vessel diseases such as hemagiomas, and capillary proliferation within
atherosclerotic
plaques; Osler-Webber Syndrome; myocardial angiogenesis; plaque
neovascularization;
telangiectasia; hemophiliac joints; angiofibroma; and wound granulation. Other
uses include
the treatment of diseases characterized by excessive or abnormal stimulation
of endothelial
3o cells, including not limited to intestinal adhesions, Crohn's disease,
atherosclerosis,
scleroderma, and hypertrophic scars, i.e., keloids. Another use is as a birth
control agent, by
inhibiting ovulation and establishment of the placenta. The compounds of the
invention are
also useful in the treatment of diseases that have angiogenesis as a
pathologic consequence
such as cat scratch disease (Rochele minutesalia qui~ztasa) and ulcers
(He~icobacter~ylori).
The compounds of the invention are also useful to reduce bleeding by
administration prior to
surgery, especially for the treatment of resectable tumors.
-9-
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Synthetic Methods
Abbreviations which have been used in the descriptions of the scheme and the
examples that follow are: BOC for tart-butoxycarbonyl; PCC for pyridinium
chlorochromate;
,EDC for 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; HOAT for
1-
hydroxy-7-azabenzotriazole; DCC for 1,3-dicyclohexylcaxbodiimide; HC?BT for 1-
hydroxybenzotriazole; DMSO for dimethylsulfoxide; BOC-ON for [2-(tert-
butoxycarbonyloxyimino)-2-phenylacetonitrile; and THF for tetrahydrofuran.
The compounds and processes of the present invention will be better understood
in
connection with the following synthetic schemes which illustrate the methods
by which the
compounds of the invention may be prepared. Starting materials can be obtained
from
commercial sources or prepared by well-established literature methods known to
those of
ordinary skill in the art. The groups Rl, and R2 are as defined above unless
otherwise noted
below.
This invention is intended to encompass compounds having formula (1~ when
prepared by synthetic processes or by metabolic processes. Preparation of the
compounds of
the invention by metabolic processes include those occurring in the human or
animal body (~32
vivo) or processes occurring in vitro.
Schegne 1
By Rt
OH ~OH
BOCHN ~ BOCNN
O
(2) (3)
p p
H2N OH
BOCHN
OH O
(Ia) (4)
2o As shown in Scheme 1, compounds of formula (2) can be converted to
compounds of
formula (3) by treatment with a reducing agent. Examples of reducing agents
include sodium
bis(2-methoxyethoxy)aluminum hydride (Red-Al~), lithium aluminum hydride, and
borane.
Oxidation of compounds of formula (3) can be accomplished by treatment with an
oxidizing
agent such as sulfur trioxide pyridine complex, PCC, or the Dess-Martin
periodinane to
provide compounds of formula (4). Conversion of compounds of formula (~) to
compounds
of formula (4) may also be achieved by conversion to the N-methoxy-N
methylamide using
N, O-dimethyIhydroxylamine and an activating agent such as DCC or EDCI,
followed by
treatment with a reducing agent such as lithium aluminum hydride. Reacting
compounds of
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formula (4) with trimethylsilylcyanide or sodium bisulfate and potassium
cyanide provides the
cyanohydxin which can be treated ira situ with a strong acid such as HCl to
simultaneously
hydrolyze the cyano group and remove the protecting group, providing compounds
of formula
(Ia) (compounds of formula (I) where R2 is hydrogen).
Scheme 2
SR3
O O
NH2 H2N
HO H H OOH OH
BOCHN
S-S
O
(6) (2a)
Compounds of formula (2a) can be prepared according to the procedure shown in
Scheme 2. Reduction of compounds of formula (6) with liquid ammonia and sodium
1o followed by treatment with an alkyl halide provides compounds of'formula
(2a) where R3 is
alkyl. Alternatively, compounds of formula (2a} can be prepared by treating
compounds of
formula (6) with liquid ammonia and sodium followed by a strong acid (such as
HCl) and an
appropriately substituted alcohol (R30H).
Scheme 3
O O BOCHN O
R1/ \H R1 ~ ORa ~ Ry OR4
{9) OH
HZN O BOCHN O
R' OH Ri OH
OH OH
15 (ta) (10)
Scheme 3 shows the synthesis of compounds of formula (Ia). Compounds of
formula
(7) can be converted to compounds of formula (8) (R4 is an alkyl group) by
treatment with the
appropriately substituted phasphonate in the presence of a base such as sodium
hydride,
potassium hydride, or lithium hexamethyldisilazide. Conversion of compounds of
formula
2o (8) to compounds of formula (9) can be accomplished by treatment with tart-
butylcarbamate,
tent-butylhypochlorite, and NaOH, followed by treatment with potassium osmate
and either
hydroquinine 1,4-phthalazinediyl diether or 1,4-phthalazinediyl diether
(depending on which
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enantiomer is desired). Hydrolysis of the ester can be accomplished by methods
known to
those of ordinary skill in the aut (for example, lithium hydroxide and
hydrogen peroxide) to
provide cornpounds of formula (10), which can be deprotected using conditions
kno~rn to
those of ordinary skill in the ant (such as HCl) to provide compounds of
formula (Ia).
Scheme 4
R~ O R~ O
BOCHN OH ~ BOCHN ORz
OH OH
(S) (11)
R~ O
NH2 ~ ~ORZ
OH
(~)
Compounds of formula (Ib) can be prepared by the methods shown in Scheme 4.
Compounds of formula (5) can be esterified by treatment with an aIkylating
agent such as
trimethylsilyldiazomethane or by coupling with the corresponding alcohol
(R20H) in the
to presence of a coupling agent such as EDC and H(~AT or DCC and HOBT to
provide
compounds of formula (11). Conversion of compounds of formula (I1) to
compounds of
formula (Ib) (compounds of formula (I) where Ra is other than hydrogen) can be
accomplished by methods known to those of ordinary skill in the art (for
example, treatment
with HCl).
Scheme 5
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R1 O
BOCHN ~ ~OR2
OH
(11)
(12)
R1 O
H2N ~ ~OR2
OH
(Ib) (14) (13)
Compounds of formula (Ib) can also be prepared by the procedures described in
Scheme 5. Compounds of formula (11) can be treated with 2,2-dirnethoxypropane
in the
presence of an acid such as methylbenzenesulfonic acid or p-toluenesulfonic
acid to provide
compounds of formula (12). Hydrolysis of the ester using conditions known to
those of
ordinary skill in the art (for example, lithium hydroxide) provides compounds
of formula
(13). Esterification of compounds of formula (13) can be accomplished by
treatment with an
appropriately substituted alcohol (R20H) and a coupling agent such as EDC and
HOAT or
DCC and HOBT. Compounds of formula (14) can be treated with a strong acid such
as HCl
l0 in the presence of water to provide compounds of formula (Ib}.
The present invention will now be described in connection with certain
preferred
embodiments which are not intended to limit its scope. On the contrary, the
present invention
covexs all alternatives, modifications, and equivalents as can be included
within the scope of
the claims. Thus, the following examples, which include preferred embodiments,
will
15 illustrate the preferred practice of the present invention, it being
understood that the examples
are for the purposes of illustration of certain preferred embodiments and are
presented to
provide what is believed to be the most useful and readily understood
description of its
procedures and conceptual aspects.
Compounds of the invention were named by ACD/ChemSketch version 5.fl
20 (developed by Advanced Chemistry Development, Tnc., Toronto, ON, Canada) or
were given
names which appeared to be consistent with ACD nomenclature.
Example 1
(2RS,3.R)-3-amino-2-h droxy-5-(methylsulfan~pentanoic acid
Example lA
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tent-butt(1R)-1-(hydroxymethyl)-3-(methylsulfanyl)propylcarbamate
A solution of N (tent-butoxycarbonyl)-D-methionine (12.47 g, 50 mmol) and
sodium
bis(2-methoxyethoxy)aluminum hydride (Red-Al~) (50 mmol) in day toluene (1~5
mL) was
stirred at 0 °C for 30 minutes, then at ambient temperature for 1 hour.
The mixture was
treated with aqueous Rochelle salt and extracted with diethyl ether. The
extract was washed
sequentially with brine and aqueous NaHC03, dried (MgSO~.), filtered, and
concentrated to
provide the desired product (9.05 g).
Example 1B
1o tert-butyl (1R)-1-formyl-3-(meth lsulfanyl)~ropylcarbamate
A solution of Example lA (9.05 g, 38.5 mmol), sulfur trioxide pyridine complex
(30.64 g,192.5 mmol), and triethylamine (26.8 mL, 192.5 mmol) in DMSO (30 mL)
was
stirred at ambient temperature for 30 minutes, cooled to 0 °C, treated
sequentially with water
(20 xnL) and saturated aqueous KHSOq. (120 mL), and extracted with ethyl
acetate. The
I5 extract was washed sequentially with saturated aqueous KHS04 and brine,
dried (MgS04),
filtered, and concentrated to provide the desired product (9.00 g).
Example 1C
2RS,3R)-3-amino-2-hydrox~5-(methylsulfanyl)pentanoie acid
2o A solution of Example 1B (9.00 g, 38.5 mmol) and sodium bisulfate (3.80 g,
36.6
mrnol) in water (200 rnL) was stirred at 5 °C for 72 hours, warmed to
ambient temperature,
treated with a mixture of potassium cyanide (2.51 g, 38.6 mmol) in ethyl
acetate (250 mL),
and stirred for 4 hours. The separated ethyl acetate layer was washed
sequentially with water
and brine, dried (MgS04), filtered, and concentrated. The concentrate was
dissolved in
25 dioxane (75 mL) and 12M HCl~(75 mL), heated to reflux for 16 hours,
concentrated,
dissolved in water (8 imL) and acetone (300 mL), adjusted to pH 5.5 with 1M
NaOH, and
filtered. The filter cake was dried under vacuum to provide the desired
product (5.81 g).
MS (ESI(+)Q1MS) m/e 180 (M+H)+, 202 (M+ Na)~; MS (ESI(-)) m!e 178 (M-H) ;1H
NMR
(300 MHz, D20) 8 4.25 (d, O.SH), 4.14 (d, 4.5H), 3.78 (m, O.SH), 3.66 (m,
0.5H), 2.65 (m,
30 2H), 2.13 (s, 1.5H), 2.09 (s, 1.5H), 1.93 (m, 2IT).
Example 2
(2RS,3R)-3-amino-5-(eth Isu~IfanyI)-2-h~xy~entanoic acid
35 Example 2A
(2RSi3R)-3-f(teat-butoxycarbonYl)aminol-5-(eth ls~ ulfanyI)-2-hydroxypentanoic
acid
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The desired product was prepared by substituting (2R)-2-((tert-
butoxycarbonyl)amino)-4-(ethylsulfanyl)butanoic acid for (2R)-2-((tert-
butoxycarbonyl)amino)-3-cyclohexylpropanoic acid in Examples 3A-3C. MS (ESI)
mle 294
(M+H)+.
Exam In a 2B
(2RS,3R)-3-amino-5-(ethylsulfan~l -2-hydroxypentanoic acid
The desired product was prepared by substituting Example 2A for Example 3C in
Example 3D. MS (ESI] mle 194 (M+H)+;1H NMR (300 MHz, DMSO-d6) S 8.14 (br s,
l0 0.6H), 7.95 (br s, 1.4H), 4.35 (d, 0.3H), 4.16 (d, 0.7H), 3.70 (m, 0.3H),
3.46 (m, 0.7H), 2.63
(m, 2I~, 2.49 (m, 2H), 1.84 (m, 2H), 1.18 (m, 3H).
Example 3
(2RS,3R)-3-amino-4-cyclohexyl-2-hydroxybutanoic acid
Exam lp a 3A
tef~-butyl (1R)-2-cyclohexyl-1-(hydroxymeth~rl)eth~lcarbamate
A solution of (2R)-2-((tent-butoxycarbonyl)amino)-3-cyclohexylpropanoic acid
(30.4
g,112 mmol) in toluene (300 mL) at 0 °C was treated with sodium bis(2-
2o rnethoxyethoxy)aluminum hyd~~de (Red-AI~) (115 mmol) over 45 minutes. The
mixture was
stirred for 30 minutes, warmed to room temperature, stirred for 1 hour,
treated with aqueous
Rochelle salt, and extracted with diethyl ether. The extract was washed
sequentially with
brine and aqueous NaHC03, dried (MgSO4), filtered, and concentrated to provide
the desired
product.
Exam lp a 3B
tent-butyl (1R)-2-c clohexyl-1-forrilylethylcarbamate
A solution of Example 3A (25.8 g,100 mmol), sulfur trioxide pyridine complex
(79.6
g, 500 mmol), and triethylamine (69.7 mL, 500 mmol) in DMSO (70 mL) at room
3o temperature was stirred for 30 minutes, cooled to 0 °C, treated with
water and saturated
aqueous KHS04, and extracted with ethyl acetate. The extract was washed
sequentially with
saturated aqueous KHS04 and brine, dried (MgS04), filtered, and concentrated
to provide the
desired product.
Example 3C
(2RS 3R)-3-f (tart-butoxycarbonyl)aminol-4-c clue 1-~2-hydrox~utanoic acid
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A solution of Example 3B (19.7 g, 77.1 mmol) and sodium bisulfate (8.0 g, 77.1
mmol) in water (500 mL) at 5 °C was stirred for 24 hours, warmed to
room temperature,
treated v~ith a solution of potassium cyanide (5.I g, 78.8 mmol) in ethyl
acetate (350 mL), and
stirred for 5 hours. The aqueous phase was separated and extracted with ethyl
acetate. The .
combined extracts were washed sequentially with water and brine, dried
(MgS04), filtered,
and concentrated. The concentrate was dissolved in dioxane (150 mL), treated
with I2M HCl
(150 mL), heated to reflux, stirred for 21 hours, and cooled to room
temperature. The
mixture was concentrated, dissolved in a mixture of water (30 mL) and acetone
(200 mL),
adjusted to pH 5.5 with 1M NaOH, treated with acetone (3.5 L), and cooled to
l0 0 °C for 4 hours. The resulting precipitate was collected by
filtration. The filter cake was
dried, dissolved in 1:1 water/dioxane, treated with BOC-ON (1.2 eq.), and
triethylamine (2
eq.), heated to 45 °C, stirred for 15 hours, treated with 10% aqueous
KHS04, and extracted
with ethyl acetate. The extract was washed sequentially with water and brine,
dried (MgSO~.),
filtered, and concentrated to provide the desired product.
Example 3D
(2RS,3R)-3-amino-4-~clohex 1-~2-hydroxybutanoic acid
A solution of Example 3C (75 mg, 0.25 mmol) in 4M HCI in dioxane (2 mL) at
room
temperature was stirred for 1 hour, concentrated, then purified by reverse
phase HPLC with
2o acetonitrile/water to provide the desired product. MS (ESI] m/e 202
(M+IT)~; 1H NMR (300
MHz, DMSO-d6) ~ 7.81 (br s, 2I-~, 4.07 (d, 1H), 3.35 (m, 1H), 1.80-1.67 (m,
5H), 1.57-1.51
(m, l~, 1.46-1.17 (m, 5H), 1.00-0.90 (m, 2H); Anal. Calcd. for CloFi2oC1NO3:
C, 50.52; H,
8.48; N, 5.89. Found: C, 50.18; H, 8.56; N, 5.67.
Example 5
f2RS,3R)-3-amino-3-cyclohexyl-2-hydroxy~ro~anoic acid
The desired product was prepared by substituting (2R)-2-((tert-
butoxycarbonyl)amino)-2-(cyclohexyl)ethanoic acid for (2R)-2-((tent-
butoxycarbonyl)amino)-
3-cyclohexylpropanoic acid in Examples 3A-3D. MS {ESn m/e 188 (M+H)+;1H NMR
(300
MHz, DMSO-d6) S 7.92 (br s, 0.6H), 7.77 (br s, 1.4H), 4.31 (d, 0.3H), 4.12 (d,
0.7H), 3.69
(m, 0.7H), 3.48 (m, 0.3H), 1.85-1.70 (m, 4H), 1.70-1.51 (m, 2H), 1.46-1.17 (m,
3H), 1.00-
0.90 (m, 2H}.
Example 6
(ZRS,3R)-3-amino-2-h~drox r-~5-pheny~entanoic acid
Example 6A
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(2RS,3R)-3-f(tert-butox ca~bonyl~aminol-2-h drox~-5-phenylpentanoic acid
The desired product was prepared by substituting (2R)-2-((tert-
butoxycarbonyl}amino)-4-(phenyl)butanoic acid for (2R)-2-((tent-
butoxycarbonyl)amino)-3-
cyclohexylpropanoic acid in Examples 3A-3C. MS {ES>] m/e 310 (M+H)+.
Example 6B
(2RS,3R)-3-amino-2-h droxy-5-nhenyl~entanoic acid
The desired compound was obtained by substituting Example 6A for Example 3C in
Example 3D. MS (ESI(+)) m/e 210 (M+H)+; 1H NMR (300MHz, DMSO-d6) b 7.34-7.16
(m,
io 5H), 4.19 (br d, 1H), 2.76-2.65 (m, 2H), 1.97-1.68 (m, 2H).
Example 7
(2RS,3R)-3-amino-2-hydro~-5-(isoprop lsulfanyl)~entanoic acid
15 Example 7A
~2R)-2-f(tert-butox cy arbonyl)aminol-4-fisoprop ls~yl)butanoic acid
A solution of D-homocystine (20 g, 75 mmol) in liquid ammonia (600 mL) was
treated sequentially with sodium (8.9 g, 390 mmol) and 2-bromopropane (20 mL,
210 mmol).
The ammonia was allowed to evaporate under a stream of nitrogen, and the
residue was
2o dissolved in 1:1 2-propanol/water (500 mL), treated with di-ten-butyl
dicarbonate (50 g, 230
mmol), stirred at room temperature for 6 hours, then concentrated. The residue
was dissolved
in water and the pH was adjusted to 10 with 50°lo NaOH. The solution
was washed twice
with diethyl ether, adjusted to pH 2 with 12M HCI, and extracted twice with
ethyl acetate.
The extracts were dried (MgSO4}, filtered, then concentrated to provide the
desired product.
25 MS (ESI) m/e 279 (M+H)+.
Example 7B
(2RS 3R)-3-f(tert-butox carbonyl)aminol-2-hydroxy-5-(isopropylsulfan~pentanoic
acid
The desired product was prepared by substituting Example 7A for (2R)-2-((tert
butoxycarbonyl)amino)-3-cyclohexylpropanoic acid in Examples 3A-3C. MS
(ESI(+}) mle
30 308 (M+H)+.
Example 7C
(2RS,3R)-3-amino-2-hydrox -5-(iso ropylsulfanyl)pentanoic acid
The desired product was prepared by substituting Example 7B for Example 3C in
35 Example 3D. MS (ESJ] m/e 208 (M+H)~; iH NMR (300 MHz, DMSO-d6) & 7.34 (br
s,
0.6H), 7.20 (br s, 1.4H), 4.10 (d, 0.3I~, 4.02 (d, 0.7H), 3.70 {m, 0.7H), 3.46
(m, 0.3H), 2.92
(m, 1H), 2.61 (m, 2H), 1.83-1.75 (m, 2H), 1.21 (d, 2.1H), 1.19 (d, 0.9H).
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Example 8
(~S 3R 4S)-3-amino-~-hydrox~4-(3-hydroxypropyl)-7-methyloctanoic acid
Example 8A
5-methylhexanoyl chloride
A mixture of 5-methylhexanoic acid (25 mL~ 175 mmol) and thionyl chloride (50
znL,
0.69 mol) was stirred at reflux for 90 minutes. The mixture was concentrated,
then distilled
at reduced pressure to give the desired product (22.35 g).
Example 8B
(4R)-4-benzyl-3-(5-meth~hexanoyl)-1 3-oxazolidin-2-one
To a -78 °C solution of 4(R)-benzyl-2-oxazolidinone (21.28 g, 0.12 mol)
in THF (360
mL) was added ~a-butyllithium (48 mL, 2.5M in hexane, 120 mmol). After 10
minutes
Example 8A (19.76 g, 0.134 mol) was added. After 30 minutes the mixture was
warmed to 0
°C, and quenched with saturated ammonium chloride. The supernatant was
decanted and
concentrated. The residue was partitioned between water and ethyl acetate and
the organic
layer was washed sequentially with water,1M sodium bicarbonate, water, and
brine, dried
(MgSO4), filtered, and concentrated to provide the desired product. MS (ESA
m/e 290
(M+~~.
Example 8C
~R~-4-benzyl-3-f (2~-2-(3-methylbutyl)-4-~entenoyll-13-oxazolidin-2-one
To a stirred -78 °C solution of Example 8B (15.65 g, 54.2 mmol) in THF
(I60 mL)
was added sodium bis(trimethylsilyl)amide (55 mL, 1M in THF). The reaction
mixture was
stirred for 45 minutes and a11y1 bromide (5.3 mL, 60.9 mmol) was added. The
reaction
mixture was stirred for 90 minutes, warmed to 0 °C, and stirred an
additional 90 minutes.
The reaction was quenched with saturated ammonium chloride, concentrated, and
partitioned
between water and ethyl acetate. The organic layer was washed sequentially
with water, 1M
3o sodium bicarbonate, water, and brine, dried (MgS04), filtered, and
concentrated to provide
the desired product. MS (ESZ] m/e 330 (M+I~~.
Example 8D
~25~' -2-(3-methylbutyl)-4-pentenoic acid
To a 0 °C solution of Example 8C (17.3 g, 52.6 mmol) in THF (200 mL)
was added
water (25 mL) and 30°Io hydrogen peroxide (25 mL, 220 mmol). A solution
of lithium
hydroxide monohydrate {4.38 g, 104 mmol) in water (100 mL) was added and the
resulting
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solution was stirred for 12 hours. ~l rie mixture was quenched with aqueous
NaHS03 then
concentrated and made basic with 5M NaOH. The aqueous phase was washed with
ethyl
acetate, then acidified with concentrated HCI to pH 3 and extracted twice with
ethyl acetate.
The combined extracts were dried (MgS04), filtered, and concentrated to
provide the desired
product. MS (ESI(+)) mle 171 (M+H)+.
Example 8E
(2S)-N methoxy-N-methyl-2-(3-meth~lbut 1~4-pentenamide
A solution of Example 8D (4.54 g, 26.7 mmol), N, O-dimethyl hydroxylamine
l0 hydrochloride (5.6 g, 57 mmol), 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide
hydrochloride (5.19 g, 27 mmol), 1-hydroxybenzotriazole (4.I2 g, 30.5 mmol),
and N
methylmorpholine (7.0 mL, 64 mmol) in dichloromethane (270 mL) at room
temperature was
stirred for 4 hours, diluted with dichlor~methane, washed sequentially with
aqueous
NaHC03, brine, 10% KHS04, and brine, dried (MgSO~.), filtered, and
concentrated. Flash
15 column chromatography (silica gel, 10% ethyl acetate/hexanes) provided the
desired product.
MS (ESI(+)) m/e 214 (M+H)~.
Example 8F
(2S)-2-(3-hydroxypropyl)-N methoxy-N,5-dimethylhexanamide
2o A solution of Example 8E (2.31 g, 10.8 mmol), and THF-borane complex (12.0
mL,
1.OM in THF) in THF (10 mL) at 0 °C was stirred for 2.5 hours, treated
with 30% aqueous
hydrogen peroxide (I0 mL, 88 mmol) and pH 7 buffer solution (20 mL), stirred
vigorously
for 90 minutes, diluted with dichloromethane, washed with pH 7 buffer, dried
{Na2S04),
filtered, and concentrated to provide the desired product. MS (ESI] m/e 232
(M+H)+.
Exam l~p a 8G
~2S)-2-(3-1 ~tert-butyl(dimethyl)silyllox~propyl)-N methoxy-N 5-
dimethylhexanamide
A solution of Example 8F (2.58 g, 11.1 mmol), tent-butyldimethylsilyl
trifluoromethanesulfonate (2.8 mL, 12 mmol), and 2,6-lutidine (1.5 mL, 13
mmol) in
3o dichloromethane (100 mL) at 0 °C was stirred for 2 hours; diluted
with dichloromethane,
washed sequentially with IO% KHS04, aqueous NaHC03, and brine, dried (MgS04),
filtered,
and concentrated to provide the desired product. MS (ESl~ m/e 346 (M+H)+.
Example 8H
~2S)-2-(3-~ ftert-butyl(dimeth 1)sil~loxylpropyl)-5-methylhexanal
A solution of Example 8F (3.16 g, 9.16 mmol) and lithium aluminum hydride
(0.34 g,
9.2 mmol) in diethyl ether (100 mL) at room temperature was stirred 30
minutes, treated with
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1M NaHS04, diluted with diethyl ether, washed sequentially with 10% NaHSO~,
and brine,
dried (MgS04), filtered, and concentrated to provide the desired product. MS
(ESI) n~le 287
(M+H)~.
Example SI
benzyl (2E,4S)-4-(3-l ftert-butyl(dimethyl)silylloxy)pro~yl)-7-meth-2-
octenoate
A solution of Example 8H (2.6I g, 9.I6 mmol), benzyl diethylphosphonoacetate
(2.83
g, 9.90 mmol), and sodium hydride (0.24 g, IO mmoI) in benzene (40 mL) at room
temperature was stirred 2 hours, diluted with ethyl acetate, washed
sequentially with saturated
l0 aqueous NH4C1 and pH 7 buffer, dried (MgSO~), filtered, and concentrated.
Flash column
chromatography (silica gel, 2:1 hexanes/CH2C12) provided the desired product.
MS (ESI) mle
419 (M+H)+.
Example 8J
15 benzyl (2S,3R,4,S7-3-f(tent-butoxycarbonyl)aminol-4-(3-dftert-
butyl(dimeth 1)siIylloxylprop~l)-2-l~droxy-7-methyloctanoate
The desired product was prepared by substituting Example 8I for Example 9B in
Example 9C. Flash column chromatography (silica gel, 10% ethyl
acetate/hexanes) provided
the purified product. MS (ESTj mle 552 (M+H)+.
Exam 1P a 8I~
(2S,3R,45~-3-amino-2-h'rdrox -~4-(3-h~oxypropyl)-7-meth~loctanoic acid
A mixture of Example 8J (15 mg, 0.027 mmol) and 10% PdlC in THF (5 mL) under a
hydrogen atmosphere at room temperature was stirred for 3.5 hours and
filtered. The filtrate
2S was treated with tetrabutylammonium fluoride (0.25 mL, 1.OM in TIC for 18
hours, diluted
with diethyl ether, washed twice with 1M HCI, dried (Na25O4), filtered, and
concentrated.
The concentrate was dissolved in 4M HGl in dioxane (2 mL), stirred for 1 hour,
and
concentrated to provide the desired product. MS (ESI) m/e 248 (M+H)~; 1H NMR
(300
MHz, DMSO-d6) ~ 4.21 (d, 1H), 3.73 (m, 1H), 3.67 (rn, 1H), 3.56 (m, 1H), 1.60-
1.48 (m,
4H), 1.43-1.37 (m, 2H), 1.35-1.23 (m, 4H), 0.91 (d, 6H).
Example 9
2S,3R)-3-amino-2-hydroxy-6-phenylhexanoic acid
Example 9A
4- henylbutanal
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A solution of 4-phenylbutyric acid (1.64 g, 10.0 mmol), N,O-dimethyl
hydroxylamine
hydrochloride (1.58 g, 16 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (2.06 g, 10.7 mmol), 1-hydroxybenzotriazole (1.56 g, 11.6 mmol),
and N-
methylmorpholine (2.8 mL, 26 mmol) in dichloromethane (40 mL) at room
temperature was
stirred for 16 hours, diluted with dichloromethane, washed sequentially with
aqueous
NaHC03, brine, 10% KHSO~., and brine, dried (MgS04), filtered, and
concentrated. A
mixture of the concentrate and lithium aluminum hydride (9.0 mmol, 1 equiv.)
in diethyl
ether (49 mL) at room temperature was stirred for 90 minutes, treated with 1M
NaHS04,
diluted with diethyl ether, washed sequentially with 10% KHS04, and brine,
dried (MgS04),
l0 filtered, and concentrated to provide the desired product. MS (ESI) m/e 148
(M+H)+.
Exam lp a 9B
ethyl (2E)-6-phenyl-2-hexenoate
A solution of Example 9A (1.17 g, 7.9 mmol), triethyl phosphonoacetate (1.05
mL,
7.9 mmol), lithium bromide (0.70 g, 8.1 mmol), and triethylamine (1.1 mL, 7.8
mmol) in
THF (80 mL) at room temperature was stirred for 16 hours, quenched with water,
stirred for
15 minutes, diluted with ethyl acetate, washed sequentially with pH 7 buffer
and brine, dried
(MgS04), filtered, and concentrated. The concentrate was purified by flash
column
chromatography on silica gel with 1:1/dichloromethane:hexanes to provide the
desired
product. MS (ESI) mle 218 (M+H)+.
Example 9C
ethyl (2S,3R)-3-f(tef~-butoxycarbonyl)anv.nol-2-h dery-6-phenylhexanoate
A solution of tef-t-butylcarbamate (1.00 g, 8.55 mmol), tart-butylhypochlorite
(0.96
mL, 8.5 mmol}, and 0.5M NaOH (17 mL, 8.5 mmol) in 1-propanol (30 mL) at room
temperature was stirred for 15 minutes. Example 9B (0.614 g, 2.82 mmol),
potassium osrnate
dihydrate (0.030 g, 0.08 mmol) and hydroquinine 1,4-phthalazinediyl diether
(0.109 g, 0.14
mmol) were added, and the mixture was stirred at 0 °C for 2 hours,
diluted with ethyl acetate,
washed sequentially with water, 1M HCl, aqueous NaHC03, and brine, dried
(MgSO~.),
3o filtered, and concentrated. The concentrate was purified by flash column
chromatography on
silica gel with 1:4/ethyl acetate:hexanes to provide the desired product. MS
(ESI) m/e 352
(M+H)+.
Exam Ip a 9D
(2S 3R)-3-f (tart-butoxycarbonyl)aminol-2-h~y-6-phenylhexanoic acid
A solution of Example 9C (0.316 g, 0.90 mmol), 30% hydrogen peroxide (0.40 mL,
3.5 mmol), and lithium hydroxide monohydrate (0.076 g, 1.8 mmol) in 3:1
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tetrahydrofuranlwater (8 mL) was stirred at 0 °C for 3 hours and
concentrated. The
concentrate was dissolved in water and the pH adjusted to 10 with 1M NaOH. The
solution
was washed twice with diethyl ether, adjusted to pH 2 with 1M HCl, and
extracted twice with
ethyl acetate. The combined extracts were dried (MgS04), filtered, and
concentrated ~to
provide the desired product. MS (ESI] m/e 324 (M+H)~.
Example 9E
(2S,3R)-3-amino-2-hydroxy-6-t~henylhexanoic acid
A solution of Example 9D (25.7 mg, 0.08 mmol) in 4M HCl in dioxane (2 mL) was
stirred at room temperature for 1 hour and concentrated to provide the desired
product. MS
(ESI] m/e 224 (M+H)~; IH NMR (300 MHz, DMSO-d~) S 7.29 (rn, 2H), 7.18 (m, 3H),
4.11
(d, 1H), 3.69 (m, 1H), 2.57 (m, 2H), 1.73-1.49 (m, 4H).
Exam lp a 10
(2RS,3R)-3-amino-5-(tart-butylsulfanyl)-2-hydroxypentanoic acid
Example l0A
(2R)-2-f(tern-butoxycarbonyl)aminol-4-(tart bu~lsulfanyl)butanoic acid
A solution of D-homocystine (3.0 g, l l mmol) in liquid ammonia (I00 mL) was
2o treated with sodium (1.3 g, 56 mmol). The ammonia was allowed to evaporate
under a
stream of nitrogen, and the residue was dissolved in 1M HCl (10 mL). The pH
was adjusted
to 0 with concentrated HCI, treated with tart-butanol (5 mL, S3 mmol), heated
to reflux for 16
hours, and concentrated. The concentrate was dissolved in 1:1 2-propanollwater
(80 mT.~),
treated with di-tart-butyl dicarbonate (20.2 g, 92.7 mmol), stirred at room
temperature for 16
hours, and concentrated. The concentrate was dissolved in water adjusted to pH
10 with 50%
NaOH. The solution was washed twice with diethyl ether, adjusted to pH 2 with
HCI, and
extracted twice with ethyl acetate. The combined extracts were dried (MgS04),
f ltered, then
concentrated to provide the desired product. MS (ESI) m/e 292 (IV!+H)~.
3o Example lOB
(2RS,3R)-3-amino-5-(tent-butylsulfan Iy )J2-hydroxypentanoic acid
The desired product was prepared by substituting Example l0A for (2R)-2-((tert-
butoxycarbonyl)amino)-3-cyclohexylpropanoic acid in Examples 3A-3D. MS (ESn
m/e 222
(M+I~~; IH NMR (300 MHz, DMSO-d6) S 4.38 (d, 0.5H), 4.28 (d, 5H), 3.73-3.66
(m, 1H),
2.72-2.63 (m, 2H), 1.95-1.84 (m, 2H), 1.34 (s, 4.5H), 1.32 (s, 2.5H).
Example 11
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methyl (3R)-3-amino-2-h droxy-5-(methylsulfan~pentanoate
Exam lp a 11A
(3R)-3-f(tent-butoxycarbonyl)aminol-2-h~oxy-5-(methylsuIfanyl)pentanoic acid
A solution of Example 1C (2.68 g, 12.5 mmol), triethylamine (3.47 ml, 75.0
mmol),
and 2-(tent-butyoxycarbonyloxyimino)-2-phenylacetonitrile (3.22 g, 13.0 mmol)
in 30%
aqueous dioxane was heated to 50 °C for 6 hours, cooled to ambient
temperature, and
concentrated. The residue was diluted with aqueous sodium hydroxide (0.25M, 50
mL) and
extracted twice with ethyl acetate. The aqueous layer was acidified to pH 3
with IM H3P04
to and extracted five times with chloroform. The combined extracts were washed
with brine,
dried (MgS04), filtered, and concentrated to provide the desired product (1.62
g). MS (ESI]
m/e 27 8 (M-H)-.
Example I lB
(3R)-3-r(tert butoxycarbonyl)aminol-5-(methylsulfanyl)-2-(tetrahydro-2H pyran-
2-
loxy)pentanoic acid
A solution of Example 11A (1.62 g, 5.8 mmol), 3,4-dihydro-2FI-pyran (0.64 mL,
70
mmol), and pyridinium p-toluenesulfonate (0.15 mL, 0.60 mmol) in
dichloromethane (30 mL)
was heated to 35 °C for 16 hours and concentrated. The concentrate was
diluted with
aqueous sodium hydroxide (0.25M, 15 mL) and extracted twice with ethyl
acetate. The
aqueous layer was adjusted to pH 3 with aqueous H3P04 (1.OM) and extracted
three times
with ethyl acetate. These three extracts were combined, washed sequentially
with water (20
mL) and brine (20 mL), dried (MgS04), filtered, and concentrated to provide
the desired
product (0.6I g). MS (APCI(+)) m/e 364. (M+H)+
Example I1C
methyl (3R)-3-((tart-butoxycarbonyl)amino)-~-h droxy-5-(methylthio)pentanoate
A solution of Example 11B (0.25g, 0.68 mmol) and trimethylsilyldiazomethane
(2.OM
in hexanes, 15 mL) in THF at room temperature was stirred for 4 hours, treated
with acetic
acid (1 mL), and concentrated. The residue was dissolved in methanol, treated
with p-
toluenesulfonic acid (10 mg), and heated to 55 °C for 16 hours. The
mixture was
concentrated and purified by flash column chromatography on silica gel with
25% ethyl
acetate/hexanes to provide the desired product. MS (APCI(+)) m/e 294 (M+I~t.
Example 11D
methyl (3R)-3-amino-2-h d~ roxy-5-(methylsulfa~l)pentanoate
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A solution of Example 11C in 4M HCI in dioxane (10 mL) at room temperature was
stirred for 4 hours and concentrated to provide the desired product. MS
(APCI(+)) m/e 194
(M-H) ; 1H NMR (300 MHz, DMSO-d6) 8 8.09 (br s, 2H), 7.94 (br s, 1H), 6.58 (m,
O.SH),
6.42 (m, 0.5H), 4.48 (m, 0.5H), 4.30 (m, 0.5H), 3.72 (s, 3H), 2.58 (m, 2H),
2.04 (s, 1.SH),
2.02 (s, I.5H), 1.82 (m, 2H).
Example 12
methyl (2RS,3R)-3-amino~4-cyclohexyl-2-h~oxybutanoate
to Example 12A
methyl (2RS,3R)-3-(tart-butoxycarbonyl)amino-4-cyclohexyl-2-h droxybutanoate
A solution of Example 3C (10.234 g, 34 mmol), and trimethylsilyldiazomethane
(2.0M in hexanes, 25 mL) in 3.5:1 benzene/methanol (232 mL) at room
temperature was
stirred at 0 °C for I hour, warmed to ambient temperature, stirred for
1 hour, quenched by the
dropwise addition of acetic acid, concentrated to provide the desired product.
MS (ES'1) mle
316 (M+H)+.
Example 12B
methyl (2RS,3R)-3-amino-4-cyclohexyl-2-h droxybutanoate
20. The desired product was prepared by substituting Example 12A for Example
3C in
Example 3D. MS (ES17 mle 216 (M+H)~; 1H NMR (300 MHz, CD30D) 8 4.42 (d, 0.3H),
4.22 (d, 0.7H), 3.80 (s, 2.1H), 3.77 (s, 0.9H), 3.~4 (m, 1H), 1.80-1.67 (m,
5H), 1.57-1.51 (m,
1H), 1.46-1.I7 (m, 5H), 1.00-0.90 (m, 2H).
2s Exam Ip a 13
(IS,2R)-2-amino-2,3-dihydro-1H inden-1-yl (2RS 3R)-3-amino-4-cyclohex
hydroxybutanoate
A solution of Example 3C (0.10 g, 0.33 mmol), (IS,2R)-1-amino-2-hydroxyindane
(0.086 g, 0.58 mmol), 1-(3-dimethylaxninopropyl)-3-ethylcarbodiimide
hydrochloride (0.080
3o g, 0.41 mmol), 1-hydroxybenzotriazole (0.061 g, 0.45 mmol), and N
methylmorpholine (0.05
mL , 0.46 mmol) in 3:1 dichloromethane/DMF (4 mL) at room temperature was
stirred for 16
hours, diluted with dichloromethane, washed sequentially with aqueous NaHC03,
brine, 10%
KHSO4, and brine, dried (MgS04), filtered, and concentrated. The residue was
dissolved in
4M HCl in dioxane (3 mL), stirred for I hour, concentrated, then purified by
reverse phase
35 HPLC with acetonitrile/water to provide the desired product. MS (ESl~ m/e
333 (M+I~+; 1H
NMR (300 MHz, DMSO-d6) 8 7.81 (br s, 2H), 7.40-7.17 (m, 4H), 6.65 (d, 1H),
5.52 (m, 1H),
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4.18 (d, 1H), 3.14 (m, 1H), 2.55-2.45 (m, 2H), 1.80-1.67 (m, 5H), 1.57-1.51
(m, 1H), 1.46-
1.17 (m, 5H), 1.00-0.90 (m, 2H).
Example 14
benzyl (2S,3R)-3-amino-3-c~pent~2-h droxypropanoate
Example 14A
bend (2E~-3-c~pentylacrylate
A solution of cyclopentanecaboxaldehyde (0.069 g, 0.70 mmol), benzyl
(diethylethyl
io phosphono)acetate (0.100 g, 0.35 mmol), lithium bromide (0.030 g, 0.35
nunol), and
triethylamine (0.049 mL, 0.35 mmol) in THF (1 mL) at room temperature was
stirred for 16
hours, quenched with water, stirred for 15 minutes, diluted with ethyl
acetate, washed
sequentially with pH 7 buffer and brine, dried (MgS04), filtered, and
concentrated to provide
the desired product. MS (ESI] m/e 231 (M+H)+.
Example 14B
benzyl (2S,3R)-3-f (tart-butox cy arbon~)aminol-3-cyclopentyl-2-h~ypropanoate
The desired product was prepared by substituting Example 14A for Example 9B in
Example 9C. MS (ESI) m/e 364 (M+H)+.
Example 14C
benzyl (2S,3R)-3-amino-3-cyclopent~ydroxypropanoate
The desired product was prepared by substituting Example 14B fox Example 3C in
Example 3D. MS (ESl] mle 264 {M+H)+; 1H NMR (300 MHz, CD30D) b 7.39 (m, 5H),
5.29
(dd, 2H), 4.04 (d, 1H), 3.88 (dd, 1H), 3.6'6 (s, 1H), 1.96 (m, 1H), 1.86 (m,
1H), 1.68 (m, 2H),
1.58 (m, 2H), 1.43 (m, IH), 1.23 (m, 1H).
Example 15
benzyl (2S,3R)-3-amino-3-cycloheptyl-2-h~ypropanoate
3o The desired product was grepared by substituting cycloheptanecarboxaldehyde
for
cyclopentanecarboxaldehyde in Examples 14A-C. MS (ES>:) m/e 292 (M+H)+;1H NMR
(300
MHz, CD30D) 8 7.39 (m, 5H), 5.29 (dd, 2H), 4.09 (d, 1H), 3.78 (dd, 1H), 3.66
(s, 1H), 1.84
(m, 1H), 1.65 (m, 2H), 1.57 (m, 2H), 1.52 (m, 2H), 1.46 (m, 2H), 1.41 (m, 2H),
1.33 (m, 2H).
Exam lp a 16
12,t'1-~-(methylamino)-2-(1-naphthyl)ethyl (2S 3R)-3-amino-5-(eth lsy ulfanyl)-
2
h droxypentanoate
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Example 16A
teat-butyl (IR)-2-hydroxy-1-(1-naphth l~ l(iyethyl)carbamate and ter-t-butyl
(1RS)-2
h~droxy-1-methyl-1-(1-naphthyl)ethyl(methyl)carbamate
A mixture of sodium hydride (dispersion in 60% mineral oiI , 440 mg, 10.9
mmol)
and methyl (2R)-[{tert-butoxycarbonyl)amino](1-naphthyl)acetate (1.43 g, 4.54
mmol) in
THF (20 mL) was stirred at 0 °C for 15 minutes, treated with methyl
iodide (1.978 mL, 31.8
mmol), stirred at 0 °C for 2 hours, warmed to room temperature, and
stirred for 16 hours.
The mixture was diluted with ethyl acetate, washed sequentially with 10%
aqueous KHS04
and brine, dried (MgS04), filtered, and concentrated to provide a mixture of
mono- and di-
methylated product (ratio: 4:1). The mixture (2.0 g) was dissolved in THF (20
mL), cooled to
0 °C, treated with sodium bis(2-methoxyethoxy)aluminum hydride (Red-
Al~) {6 mL), stirred
for 1 hour, warmed to room temperature, stirred for 1 hour, quenched with
aqueous Na2S04
(10 mL), and extracted three times with ethyl acetate. The combined extracts
were washed
twice with brine, twice with 10% KHS04, once more with brine, twice with IO%
sodium
hydrogen carbonate, three additional times with brine, dried (MgSO~.),
filtered, and
concentrated. The concentrate was purified by flash column chromatography on
silica gel
with 10% ethyl acetate/hexanes to provide the desired products MS(ESI(+)) m!e
302 (M+H)+
and 316 (M+H)+.
Example I6B
~2R)-2-(meth~amino)-2-(1-nanhthyl)ethyl (2S,3R~3-amino-5-(ethylsulfanyl)-2
hydroxypentanoate
The desired product was prepared by substituting tert-butyl (1R)-2-hydroxy-I-
(1
naphthyl)ethyl(methyl)carbamate from Example 16A and Example 2A for (IS,2R)-I-
amino-
2-hydroxyindane and Example 3C, respectively, in Example 13. The crude product
was
purified by reverse phase HPLC with acetonitrile/water to provide the desired
product. MS
(ESI(+)) m/e 377 (M+I-~~; 1H-NMR (300MHz, DMSO-d6) 8 8.29-8.19 (m, 1H), 8.06-
8.00
(m, 3H), 7.71-7.58 (m, 3H), 6.48 (d, 1H), 5.60 (br s, 1H), 4.75-4.66 (m, 1H),
4.60-4.53 (m,
21~, 3.72-3.56 (m, 1H), 2.60 (br s, 3H), 2.48-2.26 (m, 3H, includes 2.33 (q)),
1.77-1.44 (m,
2I~, 1.05 (t, 3I~.
Example I7
(2R~2-(methylamino)-2-(1-naphth ly )ethyl (2S 3R)-3-amino-5-(ethylsulfanyl)-2
h droxyperztanoate
The desired compound u~as obtained by substituting methyl (2S)-[(tert
butoxycarbonyl)amino](1-naphthyl)acetate for methyl (2R)-[(te3~'-
butoxycarbonyl)amino](1-
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naphthyl)acetate in Examples 16A-B. MS (ESI(+)) mle 377 (M+H)t; 'H-NMR
(300MHz,
DMSO-d~) 8 8.32-8.25 (m, 1H), 8.08-7.99 (m, 3H), 7.72-7.54 (m, 3H), 6.63 (d,
1H), 5.62 (br
s, 1H), 4.71-4.55 (m, 2H), 4.45-4.40 (m, 1H), 3.73-3.64 (m, 1H), 2.62 (br s,
3H), 2.53-2.42
(m, 3H, overlapped with solvent peaks), 1.94-1.70 (m, 2H), I.17 (t, 3H).
Example 18
2-(methylamino)-2-(1-na~ht~l)propyl (3R)-3-amino-5-(eth~sulfan
l~ydroxypentanoate
The desired product was prepared by substituting ter t-butyl (IRS)-2-hydroxy-1
methyl-1-(1-naphthyl)ethyl(methyl)carbamate from Example 16A and Example 2A
for
to (1S,2R)-1-amino-2-hydroxyindane and Example 3C, respectively, in Example
13. MS
(ESI(+)) m/e 391 (M+H]~; 1H-NMR (300MHz, DMSO-d6) 8 8.I5-8.03 (br s, 3I-~,
7.93-7.82
(br s, 3H), 7,66-7.56 (br s, 1H), 6.54 (d, 0.7H}, 6.45 (d, 0.3H), 4.25-4.17
(br s, 0.3H), 4.13-
4.05 (br s, 0.7H), 3.54-3.40 (m, 1H), 2.60 (br s, 3H), 2.67-2.43 (m, includes
solvent peaks),
I.93-1.59 (m, 2H), I.2I-I.07 (m, 6H).
z5
Example I9
(2R)-2-~methylamino -2-) (I-naphthyl)ethyl (2S,3R)-3-amino-4-c~clohexyl-2
h droxybutanoate
The desired product was prepaxed by substituting tef-t-butyl (1R)-2-hydroxy-1-
(1-
2o naphthyl)ethyl(methyl)carbamate from Example 16A for (1S,2R)-1-amino-2-
hydroxyindane
in Example 13. The crude product was purified by reverse phase HPLC with
acetonitxile/watex to provide the desired product. MS (ESI) m/e 385 (M+H)+; 1H
NMR (300
MHz, DMSO-d6) 8 I0.I7 (br s,lH}, 9.8 (6x s, 1H), 8.28 (d, 1H), 8.08-7.95 (m,
4H}, 7.7-7.55
(m, 3H), 6.56 (d, IH), 5.63 (br s, IH), 4.69 (dd, 1H), 4.57 (dd, 1H), 4.3 (m,
1H), 3.5 (br s,
25 IH), 2.6 (s, 3H), I.7-I.5 (m, 6H), 1.45-1.35 (m, 3H), I.22-1.05 (m, 3H),
0.88-0.73 (m, 1H).
Example 20
(2R)-2-(methylamino)-2-(1-naphthyl)ethyl (2S,3R)-3-amino-2-h droxy-5-
phenylpentanoate
The desired compound was obtained by substituting ten-butyl (1R)-2-hydroxy-1-
(1-
3o naphthyl)ethyl(methyl)carbamate from Example I6A and Example 6A for (IS,2R)-
1-amino-
2-hydroxyindane and Example 3C, respectively, in Example 13. The crude pxoduct
was
purified by reverse phase HPLC to provide the desired product. MS (ESI) mle
393 (M+H)+;
1H NMR (300 MHz, DMSO-d6} S 10.2 (br s,lH), 9.8 (br s, IH), 8.29 (d, IH), 8.15
(br s, 2IT),
8.04 (m, 3H), 7.7-7.58 (m, 3H),7.34-7.I6 (m, 4H), 6.63 (d, 1H), 5.63 (br s,
1H), 4.7-4.56 (m,
35 2H), 4.48 (br s, Il~, 3.58 (br s, zH), 2.78-2.53 (m, 2H), 2.6 (s, 3I-~,
1.95-I.75 (m, 2H).
Exam Ip a 21
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methyl (2RS,3R)-3-amino-5-(ethylsulfan l~ydroxypentanoate
Example 21A
methyl (2RS,3R)-3-f(tent-6utoxycarbonyl)aminol-5-(eth lsulfanyl)-2-
h~droxypentanoate
The desired product was prepared by substituting Example 7B and diazomethane
for
Example 3C and trimethylsilyldiazomethane, respectively, in Example 12A.
Example 21B
methyl (2RS,3R)-3-amino-5-(ethylsulfan~-hydroxypentanoate
The desired product was prepared by substituting Example 21A for Example 3C in
Example 3D. MS (ESI] m!e 208 (M+H)+; III NMR (300 MHz, DMSO-db) S 8.15 (br s,
0.6H), 7.99 (br s, 1.4H), 6.53 (d, 0.7H~, 6.39 (d, 0.3H), 4.48 (m, 0.3ITj,
4.29 (m, 0.7H), 3.70
(s, 2.1H), 3.53 (s, 0.9H), 2.63 (m, 2IT), 2.49 (m, 2H), 1.90-1.80 (m, 2IT),
1.20-1.14 (m, 3H).
Exam Ip a 22
benzyl (2RS,3R)-3-amino-5-(ethylsuIfanyI)-2-hydroxypentanoate
Example 22A
3-tent-butyl 5-methyl (4R 5RS)-4-f2-(ethylsulfan Iy )ethyll-Z 2-dimeth~-1 3-
oxazolidine-3 5-
2o dicarboxylate
A mixture of Example 21A (1.12 g, 3.65 mmol) and catalytic 4-
methylbenzenesulfonic acid (15 mg) in 2,2-dimethoxypropane (35 mL) at ambient
temperature was stirred for 36 hours, diluted with ethyl acetate, washed
sequentially with
aqueous NaHC03 and pH 7 buffer, dried (MgS04), filtered, and concentrated.
Flash column
chromatography (silica gel, 10% acetone/hexanes) provided the desired product.
MS (ESI)
m/e 348 (M+H)+.
Exam lie 2-2B
4R.5RS)-3-(tent-butoxycarbonyl)-4- f 2-(ethylsulfanyl)ethyl)-2,2-dimethvl-1,3-
oxazolidine-5-
3o carboxylic acid
A 0 °C solution of Example 22A (0.65 g, 1.87 mmol) in THF (6 mL) was
treated with
a solution of lithium hydroxide monohydrate (0.088 g, 2.1 mmol) in water (2
mL), stirred fox
2 hours, concentrated, and diluted with wafer. The aqueous phase was washed
with diethyl
ether, adjusted to pH 4 with 0.5M citric acid, and extracted twice with ethyl
acetate. The
combined extracts were dried (Na2S0~.), filtered, and concentrated to provide
the desired
product. MS (ES>) m/e 334 (M+1~~.
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Example 22C
benzyl (2RS 3R)-3-amino-5-(eth lsulfanyl)-2-hydroxypentanoate
A solution of Example 22B (0.11 g, 0.33 mmol), benzyl alcohol (0.07 mL, 0.7
mmol),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.080 g, 0.41
mmol), HOAT
(0.045 g, 0.33 mmol), and N methylmorpholine (0.055 mL , 0.50 mmol) in 3:1
dichloromethane/DMF (4 mL) at room temperature was stirred for 16 hours,
diluted with
dichIoromethane, washed sequentially with aqueous NaHC03, brine, 0.5M citric
acid, and
brine, dried (MgS04), filtered, and concentrated. The residue was dissolved in
4M HCl in
dioxane (2 mL), stirred for 1 hour, treated with water (1 mL), stirred for 1
hour, concentrated,
to and purified by reverse phase HPLC with acetonitrilelwater to provide the
desired product.
MS (ESl] m/e 284 (M+H)+; 1H NMR (300 MHz, DMSO-d6) b 7.43-7.37 (m, 5H), 5.20
(s,
1.4H), 5.19 (m, 0.6H), 4.44 (m, 0.3H), 4.32 (m, 0.7H), 3.53 (m, 1H), 2.63 (m,
2H), 2.43 (m,
2H), 1.90-1.75 (m, 2H), 1.20-1.12 (m, 3H).
15 Example 23
butyl (2RS 3R)-3-amino-5-(eth lsu~lfanyl)-2-hydroxypentanoate
The desired product was prepared by substituting 1-butanol for benzyl alcohol
in
Example 22C. MS (ESn m/e 250 (M+H)+; 1H NMR (300 MHz, DMSO-d6) 8 7.87-7.80 (br
s, 2H), 6.53 (m, 0.7H), 6.33 (d, 0.3H), 4.40 (m, 0.3H), 4.24 (m, 0.7H), 4.12
(t, 2H), 3.53 (m,
20 1H), 2.63 (m, 2H), 2.48 (m, 2H), 1.90-1.75 (m, 2H), 2.66-1.57 (m, 2H), 1.44-
1.33 (m, 2H),
1.22-1.16 (m, 3H) 0.92 (t, 3H).
Example 24
isopropyl (2S 3R)-3-amino-2-hydroxy-5-(isoprop lsy ulfanyl)pentanoate
25 The desired product was prepared by substituting 2-propanol and Example 7B
for
(1S,2R)-1-amino-2-hydroxyindane and Example 3C, respectively, in Example 13.
The crude
product was purified by reverse phase HPLC with acetonitrile/water to provide
the desired
product. MS (ESA m/e 250 (M+H)+; 1H NMR (300 MHz, DMSO-d6) b 7.97 (br s, 2H),
6.62
(d, 1H), 5.02-4.92 (m, 1H), 4.18 (t, 1H), 3.73-3.63 (m, 1H), 3.53-3.38 (m,
2H), 2.98-2.85 (m,
30 1H), 2.7-2.55 (m~ IH), I.8 (q, 1H), 1.25 (d, 6H), 1.2 (d, 6H).
Example 25
isopropyl (2RS 3R)-3-amino-5-(eth lsy ulfan l~ydroxypentanoate
The desired product was prepared by substituting 2-propanol for benzyl alcohol
in
35 Example 22C. MS (ESA m/e 236 (M+H)+; 1H NMR (300 MHz, DMSO-d~) 8 7.73 (br
s,
2H), 4.97 (m, 1H), 4.15 (d, 1H), 3.44 (m, 1H), 2.63 (m, 2H), 2.49 (m, 2H),
1.79 (m, 2H), 1.23
(d, 6H), 1.18 (m, 3H).
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Example 26
((2,2-dimeth~lwopanoyl)oxylmethyl (2RS 3R)-3-amino-5-(ethylsulfan~?-2
hydroxypentanoate
A solution of Example 22B (0.102 g, 0.3I mmol), cesium carbonate (0.1I l g,
0.34
mmol), and chloromethyl pivalate (0.055 mL, 0.38 mmol) in DMF (3 mL) at room
temperature was stirred for 24 hours; diluted with ethyl acetate, washed
sequentially with
water, aqueous NaHC03, pH 7 buffer, and brine, dried (MgS04), filtered, and
concentrated.
The residue was dissolved in 4M HCl in dioxane (2 mL), stirred for 1 hour,
treated with water
(1 mL), stirred for 1 hour, concentrated, and purified by reverse phase HPLC
with
acetonitrile/water to provide the desired product. MS (ES)) m/e 308 (M+H)+; 1H
NMR (300
MHz, DMSO-d6) 8 7.87-7.80 (br s, 2H), 6.53 (m, 0.7H), 6.33 (d, 0.3H), 4.40 (m,
0.3H), 4.24
(m, 0.7H), 4.28 (s, 0.6H), 4.26 (s, 1.4H), 3.53 (m, 1H), 2.63 (m, 2H), 2.48
(m, 2H), 1.90-1.75
(m, 2H), 1.22-1.16 (m, 12H).
is
Example 27
sec-butyl (2RS,3R)-3-amino-S-(ethylsulfan ly )-2-hydroxypentanoate
The desired product was prepared by substituting 2-butanol for benzyl alcohol
in
Example 22C. MS (ES)] m/e 250 (M+H)+; tH NMR (300 MHz, DMSO-d6) S 7.87-7.80
(br
2o s, 2H), 6.53 (m, 0.7H), 6.33 (d, 0.3H), 4.83 (m, 1H), 4.40 (m, 0.3H), 4.24
(m, 0.7H), 3.53 (m,
1H), 2.63 (m, 2H), 2.48 (m, 2H), 1.90-1.75 (m, 2H), 1.28-1.17 (m, 7H), 0.92-
0.84 (m, 3H).
Example 28
(I,3-dioxo-I,3-dihydro-2H isoindol-2- 1)Y methyl (2RS,3R)-3-amino-5-
(ethylsulfanyI)-2-
25 h~y~entanoate
The desired product was prepared by substituting N (bromomethyl)phthalimide
for
chloromethyl pivalate in Example 26. MS (ESI) m/e~ 353 (M+H)+; 1H NMR (300
MHz,
DMSO-d6) 8 7.99-7.82 (m, 4H), 6.64 (d, 0.7H), 6.48 (d, 0.3H), 5.73 (s, 1H),
4.97 (s, 1H),
4.30 (m, 0.7H), 4.18 (m, 0.3H), 3.53 (m, 1H), 2.63 (m, 2H), 2.48 (m, 2H), 1.90-
1.75 (m, 2H),
30 1.22-1.16 (m, 3H).
Example 29
sec-but I (~2RS 3R)-3-amino-2-hYdrox -~5- isopropylsulfan~)pentanoate
35 Examt~le 29A
(4R,SRS~3-(tern-butoxycarbo~l)-4-f2-(isoprop ls~ulfanyl)ethyll-2 2-dimethyl-1
3
oxazolidine-5-carboxylic acid
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The desired product was prepared by substituting Example 7B for Example 21A in
Examples 22A and 22B.
Exam lp a 29B
sec-butyl (2RS 3R)-3-amino-2-h dy roxy-5-(isoprop ls~ ulfanyl)pentanoate
The desired product was prepared by substituting 2-butanol and Example 29A for
benzyl alcohol and Example 22B, respectively, in Example 22C. MS (ESI) m/e 264
(M+H)+;
iH NMR (300 MHz, DMSO-d6) 8 7.97 (br s, 2H), 6.60-5.55 (m, 1H), 4.83 (m, 1H),
4.18 (m,
1H), 3.73-3.63 (m, 1H), 3.53-3.38 (m, 2H), 2.98-2.85 (m, 1H), 2.70-2.55 (m,
1H), 1.87-1.78
to (m, 3H), 1.25 (m, 3H), 1.20 (d, 6H), 0.89-0.83 (m, 3H).
Example 30
L(2 2-dimeth~nropanoyl)ox l~methyl (2S,3R)-3-amino-2-hydroxy-5
isoprop ls~yl)pentanoate
15 The.desired product was prepared by substituting Example 29A for Example
22B in
Example 26. The crude product was purified by reverse phase HPLC with
acetonitrile/water
to provide the desired product. MS (ESI) m/e 322 (M+H)+;1H NMR (300 MHz, DMSO-
d6)
8 7.97-7.83 (br m, 2H), 6.73 (d, 1H), 5.78 (dd, 2H), 4.33 (t, 1H), 4.16 (m,
1H), 3.73-3.63 (m,
1H), 3.53-3.38 (m, 1H), 2.98-2.85 (m, 1H), 2.70-2.55 (m,1H), 1.87-1.78 (m,
1H), 1.20 (d,
20 6H), 1.12 (s, 9H).
Example 31
(1,3-dioxo-1 3-dihydro-2H isoindol-2- 1)m~ethyl (2S 3R)-3-amino-2-hydroxy-5
(isopropylsulfan~pentanoate
25 The desired product was prepared by substituting N
(brornomethyl)phthalimide and
Example 29A for chloromethyl pivalate and Example 22B, respectively, in
Example 26. The
crude product was purified by reverse phase HPLG with acetonitrile/water to
provide the
desired product. MS (ESI) m/e 367 (M+H)+; 1H NMR (300 MHz, DMSO-d6) S 7.97-
7.83
(m, 6H), 6.63 (d, 1H), 5.72 (s, 1H), 4.97 (s, 1H), 4.32 (t, 1H), 4.17 (m, 1H),
3.73-3.63 (m,
30 1H), 3.53-3.38 (m, 1H), 2.98-2.85 (m, 1H), 2.70-2.55 (m, 1H), 1.87-1.78 (m,
1H), 1.20 (d,
6H).
Example 32
3-oxo-1 3-dihydro-2-benzofuran-1-yl (2S 3R)-3-anuno-2-hydroxy-5-
35 (isoprop, lsy ulfanyl)pentanoate
The desired product was prepared by substituting 3-bromophthalide and Example
29A
for chloromethyl pivalate and Example 22B, respectively, in Example 26. The
crude product
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was purified by reverse phase HPLC with acetonitrile/water to provide the
desired product.
MS (ESI) m/e 340 (M+H)+; IH NMR (300 MHz, DMSO-d~) b 8.03-7.92 (m, 3H), 7.83-
7.78
(m, 2H), 7.66 (d, 1H), 6.63 (s, 1H), 4.28 (m, 1H), 4.18 (d, 1H), 3.73-3.63
(111, 1H), 3.53-3.38
(m, 1H), 2.98-2.85 (m, 1H), 2.70-2.55 (m, 1H), 1.87-1.78 (m, 1H), 1.20 (d,
6H).
It will be evident to one skilled in the art that the present invention is not
limited to
the foregoing illustrative examples, and that it can be embodied in other
specific forms
without departing from the essential attributes thereof. It is therefore
desired that the
examples be considered in all respects as illustrative and not restrictive,
reference being made
to the appended claims, rather than to the foregoing examples, and all changes
which come
within the meaning and range of equivalency of the claims are therefore
intended to be
embraced therein.
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