U.S. patent application number 14/148580 was filed with the patent office on 2014-05-22 for methods and compositions for treatment of diabetes and dyslipidemia.
This patent application is currently assigned to Kareus Therapeutics, SA. The applicant listed for this patent is Ish Khanna, Sivaram Pillarisetti. Invention is credited to Ish Khanna, Sivaram Pillarisetti.
Application Number | 20140142178 14/148580 |
Document ID | / |
Family ID | 45818296 |
Filed Date | 2014-05-22 |
United States Patent
Application |
20140142178 |
Kind Code |
A1 |
Khanna; Ish ; et
al. |
May 22, 2014 |
METHODS AND COMPOSITIONS FOR TREATMENT OF DIABETES AND
DYSLIPIDEMIA
Abstract
Novel compounds of Formula I are provided: ##STR00001## its
stereoisomers and/or pharmaceutically acceptable salts for the
treatment of diabetes and diabetes-associated dyslipidemia, wherein
R.sup.1 is selected from a group consisting of hydroxy, alkoxy,
amine, alkyl, haloalkyl, NHSO.sub.2R, or NHCOR wherein R is
selected from alkyl or cycloalkyl, NHR' wherein R' is alkyl or
cycloalkyl optionally substituted by hydroxy or alkoxy; and n.sup.1
and n.sup.2 are independently selected from 0, 1, and 2. At least
one of R.sup.3 and R.sup.4 and/or R.sup.5 and R.sup.6 form a cyclic
ring of 3-8 carbon atoms optionally containing alkyl groups, hetero
atoms, or functional groups such as O, N, SO.sub.2. Additionally,
when R.sup.3 and R.sup.4 or R.sup.5 and R.sup.6 do not form a
cyclic ring, then they may be independently selected from hydrogen,
alkyl, branched alkyl, and cycloalkyl. L.sub.1 is a linear
aliphatic chain optionally containing from 4 to 16 carbon atoms.
The chain may optionally be substituted one or more times by alkyl,
branched alkyl, cycloalkyl, or aryl. R.sup.2 is independently
selected from hydrogen, alkoxy, hydroxy, alkyl, haloalkyl,
cycloalkyl, heterocycloalkyl, heteroaryl, cyano, or COR.sup.7,
wherein R.sup.7 is selected from hydroxy, alkyl, alkoxy, or amine,
NHR', NHSO.sub.2R, or NHCOR. Y.sup.1 is oxygen or hydrogen. Y.sup.2
is optional, wherein when Y.sup.2 is present, Y.sup.1 and Y.sup.2
are hydrogen. When Y.sup.2 is not present, Y.sup.1 is a carbonyl
group.
Inventors: |
Khanna; Ish; (Alpharetta,
GA) ; Pillarisetti; Sivaram; (Norcross, GA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Khanna; Ish
Pillarisetti; Sivaram |
Alpharetta
Norcross |
GA
GA |
US
US |
|
|
Assignee: |
Kareus Therapeutics, SA
La Chaux-De-Fonds
CH
|
Family ID: |
45818296 |
Appl. No.: |
14/148580 |
Filed: |
January 6, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
13236807 |
Sep 20, 2011 |
8623897 |
|
|
14148580 |
|
|
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|
Current U.S.
Class: |
514/507 ;
560/315 |
Current CPC
Class: |
C07C 43/115 20130101;
C07C 2601/02 20170501; C07C 61/15 20130101; C07C 255/46 20130101;
C07C 233/58 20130101; A61P 3/00 20180101; C07C 2601/04 20170501;
A61P 9/00 20180101; C07C 311/51 20130101; C07C 61/12 20130101; A61P
3/08 20180101; C07C 2601/08 20170501; C07D 257/04 20130101; C07C
31/27 20130101; C07C 69/608 20130101; A61P 3/10 20180101; C07C
69/74 20130101; C07C 53/132 20130101; C07C 233/60 20130101; A61P
3/06 20180101 |
Class at
Publication: |
514/507 ;
560/315 |
International
Class: |
C07C 311/51 20060101
C07C311/51 |
Claims
1.-12. (canceled)
13. A composition having the following formula: ##STR00059## its
pharmaceutically acceptable salts, tautomers, and mixtures
thereof.
14. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and at least one compound according to claim
1.
15. A method for the treatment of insulin resistance, diabetes,
and/or cardiovascular disease in a subject, the method comprising
administering to the subject a compound having the following
formula: ##STR00060## its pharmaceutically acceptable salts,
tautomers, and mixtures thereof.
Description
[0001] This application is a continuation non-provisional
application and claims priority to U.S. Provisional Patent
Application Ser. No. 61/384,446, filed Sep. 20, 2010 and U.S.
Non-Provisional patent application Ser. No. 13/236,807, filed Sep.
20, 2011, which is incorporated by reference herein in their
entirety.
BACKGROUND OF THE INVENTION
[0002] The present invention relates to methods and compositions
for the treatment of insulin resistance, diabetes and/or
diabetes-associated dyslipidemia and cardiovascular disease. More
specifically, the present invention relates to compositions of
select cycloalkyl aliphatic carboxylic acids and derivatives for
the treatment of insulin resistance, diabetes and
diabetes-associated dyslipidemia.
SUMMARY OF THE INVENTION
[0003] In one aspect, the present invention is directed to novel
compounds of Formula I:
##STR00002##
its stereoisomers and/or pharmaceutically acceptable salts for the
treatment of diabetes and diabetes-associated dyslipidemia,
wherein: R.sup.1 is selected from a group consisting of hydroxy,
alkoxy, amine, alkyl, haloalkyl, NHSO.sub.2R, or NHCOR wherein R is
selected from alkyl or cycloalkyl. NHR' wherein R' is alkyl or
cycloalkyl optionally substituted by hydroxy or alkoxy; and n.sup.1
and n.sup.2 are independently selected from 0, 1, and 2. At least
one of R.sup.3 and R.sup.4 and/or R.sup.5 and R.sup.6 form a cyclic
ring of 3-8 carbon atoms optionally containing alkyl groups, hetero
atoms, or functional groups such as O, N, SO.sub.2. Additionally,
when R.sup.3 and R.sup.4 or R.sup.5 and R.sup.6 do not form a
cyclic ring, then they may be independently selected from hydrogen,
alkyl, branched alkyl, and cycloalkyl. L.sub.1 is a linear
aliphatic chain optionally containing from 4 to 16 carbon atoms.
The chain may optionally be substituted one or more times by alkyl,
branched alkyl, cycloalkyl, or aryl. R.sup.2 is independently
selected from hydrogen, alkoxy, hydroxy, alkyl, haloalkyl,
cycloalkyl, heterocycloalkyl, heteroaryl, cyano, or COR.sup.7,
wherein R.sup.7 is selected from hydroxy, alkyl, alkoxy, or amine,
NHR'. NHSO.sub.2R, or NHCOR. Y.sup.1 is oxygen or hydrogen. Y.sup.2
is optional, wherein when Y.sup.2 is present, Y.sup.1 and Y.sup.2
are hydrogen. When Y.sup.2 is not present, Y.sup.1 is a carbonyl
group.
[0004] In another aspect, the present invention is directed to a
compound of Formula II
##STR00003##
its stereoisomers and/or pharmaceutically acceptable salts for the
treatment of diabetes and diabetes-associated dyslipidemia, wherein
R.sup.1 and R.sup.7 are independently selected from a group
consisting of hydroxy, alkoxy, alkyl, amine. NHR' wherein R' is
alkyl or cycloalkyl optionally substituted by hydroxy or alkoxy.
NHSO.sub.2R or NHCOR, wherein R is selected from alkyl or
cycloalkyl and n.sup.1 and n.sup.2 are independently selected from
0, 1, and 2. At least one of R.sup.3 and R.sup.4 and/or R.sup.5 and
R.sup.6 form a cyclic ring of 3-8 carbon atoms optionally
containing alkyl groups, hetero atoms, or functional groups such as
O, N, SO.sub.2. Additionally, when R.sup.3 and R.sup.4 or R.sup.5
and R.sup.6 do not form a cyclic ring, then they may be
independently selected from hydrogen, alkyl, branched alkyl, and
cycloalkyl. L.sup.1 is independently a linear aliphatic chain
optionally containing from 6 to 16 carbon-atoms and L.sup.1 may
optionally be substituted one or more times by alkyl, branched
alkyl, cycloalkyl, or aryl.
[0005] In yet another aspect, the invention is directed to novel
compounds of Formula I, its stereoisomers, tautomers and/or its
pharmaceutically acceptable salt thereof as AMPK activators.
[0006] In a different aspect, the invention is directed to novel
compounds of Formula I, its stereoisomers, tautomers and/or its
pharmaceutically acceptable salt thereof as inhibitors of lipid
synthesis
[0007] In another aspect, the invention is directed to a method for
the treatment of diabetes, insulin resistance, dyslipidemia,
obesity and cardiovascular disease in a subject, which comprises
administering to the subject a therapeutically effective amount of
a compound of formula (I), its stereoisomers and/or its
pharmaceutically acceptable salt thereof.
DETAILED DESCRIPTION OF THE INVENTION
[0008] Reference will now be made in detail to the embodiments of
the invention, one or more examples of which are set forth below.
Each example is provided by way of explanation of the invention,
not a limitation of the invention. In fact, it will be apparent to
those skilled in the art that various modifications can be made in
the present invention without departing from the scope or spirit of
the invention. For instance, features illustrated or described as
part of one embodiment can be used on another embodiment to yield a
still further embodiment. Thus it is intended that the present
invention cover such modifications and variations as come within
the scope of the appended claims and their equivalents. Other
objects, features, and aspects of the present invention are
disclosed in, or are obvious from, the following detailed
description.
[0009] In one aspect, the present invention is directed to novel
compounds of Formula I:
##STR00004##
its stereoisomers and/or pharmaceutically acceptable salts for the
treatment of diabetes and diabetes-associated dyslipidemia, wherein
R.sup.1 is selected from a group consisting of hydroxy, alkoxy,
amine, alkyl, haloalkyl. NHSO.sub.2R, or NHCOR wherein R is
selected from alkyl or cycloalkyl. NHR' wherein R' is alkyl or
cycloalkyl optionally substituted by hydroxy or alkoxy; and n.sup.1
and n.sup.2 are independently selected from 0, 1, and 2. At least
one of R.sup.3 and R.sup.4 and/or R.sup.5 and R.sup.6 form a cyclic
ring of 3-8 carbon atoms optionally containing alkyl groups, hetero
atoms, or functional groups such as O, N, SO.sub.2. Additionally,
if R.sup.3 and R.sup.4 or R.sup.5 and R.sup.6 do not form a cyclic
ring, then they may be independently selected from hydrogen, alkyl,
branched alkyl, and cycloalkyl. L.sub.1 is a linear aliphatic chain
optionally containing from 4 to 16 carbon atoms. The chain may
optionally be substituted one or more times by alkyl, branched
alkyl, cycloalkyl, or aryl. R.sup.2 is independently selected from
hydrogen, alkoxy, hydroxy, alkyl, haloalkyl, cycloalkyl,
heterocycloalkyl, heteroaryl, cyano, or COR.sup.7, wherein R.sup.7
is selected from hydroxy, alkyl, alkoxy, or amine, NHR'.
NHSO.sub.2R, or NHCOR. Y.sup.1 is oxygen or hydrogen. Y.sup.2 is
optional, wherein when Y.sup.2 is present, Y.sup.1 and Y.sup.2 are
hydrogen. When Y.sup.2 is not present, Y.sup.1 is a carbonyl
group
[0010] In another aspect, the present invention is directed to a
compound of Formula II
##STR00005##
its stereoisomers and/or pharmaceutically acceptable salts for the
treatment of diabetes and diabetes-associated dyslipidemia, wherein
R.sup.1 and R.sup.7 are independently selected from a group
consisting of hydroxy, alkoxy, alkyl, amine, NHR' wherein R' is
alkyl or cycloalkyl optionally substituted by hydroxy or alkoxy,
NHSO.sub.2R or NHCOR, wherein R is selected from alkyl or
cycloalkyl and n.sup.1 and n.sup.2 are independently selected from
0, 1, and 2. At least one of R.sup.3 and R.sup.4 and/or R.sup.5 and
R.sup.6 form a cyclic ring of 3-8 carbon atoms optionally
containing alkyl groups, hetero atoms, or functional groups such as
O, N, SO.sub.2. Additionally, when R.sup.3 and R.sup.4 or R.sup.5
and R.sup.6 do not form a cyclic ring, then they may be
independently selected from hydrogen, alkyl, branched alkyl, and
cycloalkyl. L.sup.1 is independently a linear aliphatic chain
optionally containing from 6 to 16 carbon-atoms and L.sup.1 may
optionally be substituted one or more times by alkyl, branched
alkyl, cycloalkyl, or aryl.
[0011] In another aspect, the present invention is directed to a
compound of Formula III
##STR00006##
its stereoisomers and/or pharmaceutically acceptable salts for the
treatment of diabetes and diabetes-associated dyslipidemia, wherein
R.sup.1 is selected from a group consisting of hydroxy, alkoxy,
amine, alkyl, haloalkyl. NHR' wherein R' is alkyl or cycloalkyl
optionally substituted by hydroxy or alkoxy, NHSO.sub.2R or NHCOR,
wherein R is selected from alkyl or cycloalkyl; and n.sup.1 and
n.sup.2 are independently selected from 0, 1, and 2. At least one
of R.sup.3 and R.sup.4 and/or R.sup.5 and R.sup.6 form a cyclic
ring of 3-8 carbon atoms optionally containing alkyl groups, hetero
atoms, or functional groups such as O, N, SO.sub.2. Additionally,
when R.sup.3 and R.sup.4 or R.sup.5 and R.sup.6 do not form a
cyclic ring, then they may be independently selected from hydrogen,
alkyl, branched alkyl, and cycloalkyl. L.sup.1 is a linear
aliphatic chain optionally containing from 4 to 16 carbon-atoms and
L.sup.1 may optionally be substituted one or more times by alkyl,
branched alkyl, cycloalkyl, or aryl. R.sup.2 is independently
selected from hydrogen, alkoxy, hydroxy, alkyl, haloalkyl,
cycloalkyl, heterocycloalkyl, heteroaryl, cyano or COR.sup.7;
wherein R.sup.7 is selected from a group consisting of hydroxy,
alkyl, alkoxy, amine. NHR', or NHSO.sub.2R.
[0012] In another embodiment of present invention, the compound of
present invention is selected from one or more of:
##STR00007## ##STR00008##
their stereoisomers, pharmaceutically acceptable salts, tautomers,
and mixtures thereof.
[0013] In another embodiment of present invention, the compound of
present invention is selected from one or more of:
##STR00009## ##STR00010##
their stereoisomers, pharmaceutically acceptable salts, tautomers,
and mixtures thereof.
[0014] In another embodiment of present invention, the compound of
present invention is selected from one or more of:
##STR00011## ##STR00012## ##STR00013## ##STR00014##
their stereoisomers, pharmaceutically acceptable salts, tautomers,
and mixtures thereof.
[0015] In one embodiment, the invention is directed to novel
compounds of Formula (I), their stereoisomers, tautomers and/or
their pharmaceutically acceptable salts as AMPK activators
[0016] In another embodiment, the invention is directed to novel
compounds of Formula (I), their stereoisomers, tautomers and/or
pharmaceutically acceptable salts as inhibitors of lipid
synthesis.
[0017] In another embodiment, the invention is directed to a method
for the treatment of diabetes, insulin resistance, dyslipidemia,
obesity or cardiovascular disease in a subject, which comprises
administering to the subject a therapeutically effective amount of
a compound of formula (I), its stereoisomers and/or
pharmaceutically acceptable salts thereof.
[0018] In another embodiment of present invention, the compounds of
Formula I, II or III may optionally be combined with one or more
anti-diabetic or dyslipidemia drugs such as metformin, DDP-IV
inhibitor, sulfonylurea, SGLT-2 inhibitors statins, alpha
glucosidase inhibitors, and insulin.
[0019] In an embodiment of present invention, metformin combines
with mono- or dicarboxylic fatty acid of Formula I, II or III to
form salts suitable for oral delivery--
##STR00015##
wherein R'CO.sub.2H is a fatty acid of present invention of Formula
I, II or III and x is the molar ratio of fatty acid in metformin
combination and may range from about 0.5 to about 3.
[0020] In another embodiment, the present invention describes a
pharmaceutical composition containing compounds of the present
invention and one or more pharmaceutically acceptable
excipients.
[0021] In yet another embodiment, the present invention describes a
method of preventing or treating insulin resistance and diabetes in
a subject. The method involves administering to a subject an
effective amount of compound(s) of the present invention or a
composition thereof.
[0022] In another embodiment, the effective amount of a compound of
the present invention is delivered orally, sublingually or
intravenously. Other administration means known in the art are also
contemplated as useful in accordance with the present
invention.
[0023] As used herein, the following terms are defined as:
[0024] `Halogen or Halo` means fluorine, chlorine, bromine or
iodine.
[0025] `Alkyl` group means linear or branched alkyl groups.
Exemplary alkyl groups include methyl, ethyl, n-propyl, iso-propyl,
n-butyl, iso-butyl, t-butyl, n-pentyl, iso-pentyl, hexyl, heptyl,
octyl and the like. Unless otherwise specified, an alkyl group
typically has from about 1 to about 10 carbon atoms.
[0026] `Cycloalky` group means a cyclic alkyl group which may be
mono or bicyclic. Exemplary cycloalkyl groups include cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and
the like. Unless otherwise specified, a cycloalkyl group typically
has from about 3 to about 10 carbon atoms.
[0027] `Alkoxy` means an --O (alkyl) group, where alkyl is as
defined above. Exemplary alkyl groups include methoxy, ethoxy,
propoxy, butoxy, iso-propoxy, iso-butoxy, and the like. Unless
otherwise specified, an alkoxy group typically has from 1 to about
10 carbon atoms.
[0028] `Amine` refers to a primary, secondary, or tertiary amino
group. The secondary and tertiary amine may contain alkyl,
cycloalkyl or aryl substitutions. Some examples of amines include
NH.sub.2, NHMe, NMe.sub.2 NH(cyclopropyl). Unless otherwise
specified, the alkyl or cycloalkyl group on an amine typically has
from 1 to about 8 carbon atoms.
[0029] `Aryl` means an optionally substituted monocyclic or
polycyclic aromatic ring system of about 6 to about 14 carbon
atoms. Exemplary aryl groups include phenyl, naphthyl, and the
like. Unless otherwise specified, an aryl group typically has from
6 to about 14 carbon atoms.
[0030] `Heteroaryl` means an aromatic monocyclic or polycyclic ring
system of about 4 to about 10 carbon atoms, having at least one
heteroatom or hetero group selected from --O--, --N--, --S--,
--SO.sub.2, or --CO. Exemplary heteroaryl groups include one or
more of pyrazinyl, isothiazolyl, oxazolyl, pyrazolyl, pyrrolyl,
tetrazolyl, imidazolyl, triazolyl, pyridazinyl, thienopyrimidyl,
furanyl, indolyl, isoindolyl, benzo[1,3]dioxolyl,
1,3-benzoxathiole, pyrrolidine 2,4-dione, quinazolinyl, pyridyl,
thiophenyl and the like. Unless otherwise specified, a heteroaryl
group typically has from 4 to about 10 carbon atoms.
[0031] `5- to 6-member heteroaryl` is an aromatic monocyclic ring
system of 5 or 6 ring atoms, having at least one heteroatom or
hetero group selected from --O--, --N--, --S--, --SO.sub.2, or
--CO. Exemplary `5- to 6-member heteroaryl` groups include one or
more of pyrazinyl, isothiazolyl, oxazolyl, pyrazolyl, pyrrolyl,
pyridazinyl, pyridyl, thienopyrimidyl, tetrazolyl, imidazolyl,
triazolyl, furanyl and the like.
[0032] `Heterocyclyl` means a non-aromatic saturated monocyclic or
polycyclic ring system of 3 to about 10 members having at least one
heteroatom or hetero group selected from --O--, --N--, --S--,
--SO.sub.2, or --CO. Exemplary heterocyclyl groups include one or
more of pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl,
thiomorpholinyl, thiomorpholine 1,1-dioxide, oxetane,
thiazolidinyl, 1,3-dioxolanyl, 1,4-dioxanyl and the like. Unless
otherwise specified, a heterocyclyl group typically has from 3 to
about 10 carbon atoms.
[0033] `Optionally substituted` means that substitution is optional
and, therefore, it is possible for the designated atom or molecule
to be unsubstituted. In the event a substitution is desired, then
such substitution means that any number of hydrogens on the
designated atom is replaced with a selection from the indicated
group, provided that the normal valence of the designated atom is
not exceeded, and that the substitution results in a stable
compound. For example in Formula I when a substituent is keto
(i.e., .dbd.O), then 2 hydrogens on the atom are replaced.
[0034] `Salts` refers to any acid or base salt, pharmaceutically
acceptable solvates, or any complex of the compound that, when
administered to a recipient, is capable of providing (directly or
indirectly) a compound as described herein. It should be
appreciated, however, that salts that are not pharmaceutically
acceptable also lie within the scope of the invention. The
preparation of salts can be carried out using known methods.
[0035] For example, pharmaceutically acceptable salts of compounds
contemplated herein as being useful may be synthesized by
conventional chemical methods using a parent compound containing a
base or an acid functionality. Generally, such salts may be
prepared, for example, by making free acid or base forms of the
compounds and reacting with a stoichiometric quantity of the
appropriate base or acid in water or in an organic solvent, or in a
mixture of the two. Generally, non-aqueous media such as one or
more of solvents such as ether, ethyl acetate, ethanol, isopropanol
or acetonitrile may be utilized. Examples of acid addition salts
include one or more of mineral acid addition salts such as
hydrochloride, hydrobromide, hydroiodide, sulphate, phosphate, and
organic acid addition salts such as one or more of acetate,
maleate, fumarate, citrate, oxalate, succinate, tartrate, malate,
mandelate, methanesulphonate and p-toluenesulphonate. Examples of
base addition salts include one or more of inorganic salts such as
sodium, potassium, calcium, ammonium, magnesium, and lithium salts,
and organic base salts such as one or more of ethylenediamine,
ethanolamine. N,N-dialkyl-ethanolamine, triethanolamine, and basic
amino acid salts.
[0036] Contemplated derivatives are those that may improve
dissolution or increase the bioavailability of the compounds of
this invention when such compounds are administered to a subject
(e.g., by making an orally administered compound more easily
absorbed. Compounds of formula I may be amorphous,
semi-crystalline, or crystalline and may either be given as parent
compounds, its salts, and/or in solvated form. The solvate may be
part of crystalline lattice or superficially associated. It is
intended that all of these forms should be within the scope of the
present invention. Methods of solvation are generally known within
the art. Suitable solvates are pharmaceutically acceptable
solvates. In one embodiment, the solvate is a hydrate.
[0037] For ease of reference, the present invention will be
described in terms of administration to human subjects. It will be
understood, however, that such descriptions are not limited to
administration to humans, but will also include administration to
other animals unless explicitly stated otherwise.
[0038] In one embodiment compounds of Formula I are useful for the
treatment of diabetes, insulin resistance, hyperlipidemia, coronary
heart disease, atherosclerosis, diabetes, and obesity. The
compounds of Formula I may influence one or more lipid parameters
such as increasing the HDL levels, lowering plasma levels of LDL,
lowering plasma glucose, or lowering triglycerides.
[0039] Compounds of formula I, their pharmaceutically acceptable
salts, and/or solvates thereof can, therefore, be used in the
prevention and/or treatment of a disease or condition discussed
above. Pharmaceutical compositions containing a therapeutically
effective quantity of a compound of formula I, its pharmaceutically
acceptable salts, and/or solvates thereof, together with
pharmaceutically acceptable excipients, are an additional aspect of
the present invention.
[0040] The therapeutically effective quantity of compounds of
formula I, their pharmaceutically acceptable salts and/or solvates
that must be administered, and the dosage for treating a
pathological state with said compounds will depend on numerous
factors, including age, the state of the patient, the severity of
the disease, the route and frequency of administration, the
modulator compound to be used, etc.
[0041] As used herein, "therapeutically effective amount" means the
dose or amount of a compound of the present invention administered
to a subject and the frequency of administration to give some
therapeutic response. The dose or effective amount to be
administered to a subject and the frequency of administration to
the subject can be readily determined by one of ordinary skill in
the art by the use of known techniques and by observing results
obtained under analogous circumstances. In determining the
effective amount or dose, a number of factors are considered by the
attending diagnostician, including but not limited to, the potency
and duration of action of the compounds used; the nature and
severity of the illness to be treated as well as on the sex, age,
weight, general health and individual responsiveness of the subject
to be treated, and other relevant circumstances.
[0042] The phrase "therapeutically-effective" indicates the
capability of an agent or combination to prevent, or improve on the
severity of, the disorder, while avoiding adverse side effects. The
therapeutically effective compositions of the present invention may
include compounds of present invention at doses of from about 50 to
about 3000 mg.
[0043] The compounds described herein are typically administered in
admixture with one or more pharmaceutically acceptable excipients
or carriers in the form of a pharmaceutical composition. A
"composition" may contain one compound or a mixture of compounds. A
"pharmaceutical composition" is any composition useful or
potentially useful in producing physiological response in a subject
to which such pharmaceutical composition is administered.
[0044] The term "pharmaceutically acceptable," with respect to an
excipient, is used to define non-toxic substances generally
suitable for use in human or animal pharmaceutical products. The
pharmaceutical composition may be in the forms normally employed,
such as tablets, capsules, powders, syrups, solutions, suspensions,
and the like. The pharmaceutical composition may contain
flavorants, sweeteners, etc., in suitable solid or liquid carriers
or diluents, or in suitable sterile media to form injectable
solutions or suspensions. Such compositions typically contain from
about 0.1 to about 50%, in some embodiments from about 1 to about
20%, by weight of active compound, the remainder of the composition
being pharmaceutically acceptable carriers, diluents or
solvents.
[0045] Suitable pharmaceutically acceptable carriers include solid
fillers or diluents and sterile aqueous or organic solutions. The
active ingredient may be present in such pharmaceutical
compositions in the amounts sufficient to provide the desired
dosage in the range as described above. Thus, for oral
administration, the active ingredient may be combined with a
suitable solid or liquid carrier or diluent to form capsules,
tablets, powders, syrups, solutions, suspensions and the like. For
parenteral administration, the active ingredient can be combined
with sterile aqueous or organic media to form injectable solutions
or suspensions. For example, solutions in sesame or peanut oil,
aqueous propylene glycol and the like can be used, as well as
aqueous solutions of water-soluble pharmaceutically-acceptable acid
addition salts or salts with base of the compounds. Aqueous
solutions with the active ingredient dissolved in polyhydroxylated
castor oil may also be used for injectable solutions. The
injectable solutions prepared in this manner can then be
administered intravenously, intraperitoneally, subcutaneously, or
intramuscularly, with intramuscular administration being preferred
in humans.
[0046] The following examples describe exemplary embodiments of the
invention. Other embodiments within the scope of the claims herein
will be apparent to one skilled in the art from consideration of
the specification or practice of the invention as disclosed herein.
It is intended that the specification, together with the examples,
be considered to be exemplary only, with the scope and spirit of
the invention being indicated by the claims which follow the
examples.
[0047] The following outlines general synthesis of compounds of the
invention. The suggested methodologies are not intended to be
limiting. The variations of these synthetic methodologies or
methodologies reported in literature can be adopted to synthesize
molecules within the scope of invention.
##STR00016##
[0048] Scheme A outlines a generic synthesis of compounds of
Formula I. The symmetrical compounds such as formula A2 and A3 are
synthesized by. quenching the carbanion generated from cycloalkyl
carboxylic acid or its ester with alkyl dihalide. Using conditions
known in literature, selective functional group transformations of
carboxyl acid, ester gives alcohol, amides, or esters that may be
further manipulated to give ether, haloalkyl, amines, ketones, etc
(A4).
[0049] The unsymmetrical compounds (B.sup.3, B.sup.4) are
synthesized using sequential anion generation followed by
alkylation steps to give B.sup.2 and B.sup.3. Functional group
manipulations of carboxyl acid, ester as discussed above gives
alcohol, amides, esters that may be further manipulated selectively
to give ether, haloalkyl, amines, ketones etc. (B.sup.4)
[0050] Similarly, the cycloalkyl (or branched alkyl) nitrile can be
used as starting points to synthesize C.sup.2 and C.sup.3. The
selective manipulation of cyano group can give easy access to
heteroaryl systems or be hydrolyzed to acids or amides and further
elaborated per need to give C.
[0051] The functional carboxylic acid group(s) in
A.sup.3/A.sup.4/B.sup.3/B.sup.4/C.sup.3/C.sup.4 may be homologated
using Arndt-Eistert synthesis or other methodologies known in
literature. The selective functional groups transformations are
used further to give target compounds of Formula I.
EXAMPLES
[0052] An embodiment of the present invention provides preparation
of the novel compounds of formula (I) using the procedures given in
the following examples. Those skilled in the art will understand
that known variations of the conditions and processes of the
following preparative procedures can be used to prepare these
compounds. Moreover, by utilizing the procedures described in
detail, one of ordinary skill in the art can prepare additional
compounds of the present invention claimed herein.
[0053] The following acronyms, abbreviations, terms and definitions
are used throughout the reaction scheme and experimental section:
[0054] THF (tetrahydrofuran) [0055] TBME (tert-butyl methyl ether)
[0056] MeOH (methanol) [0057] Ether (diethyl ether) [0058] EtOAc
(ethyl acetate) [0059] (Carbonyldiimidazole) [0060] DCM
(Dichloromethane) [0061] DMF (N,N-dimethylformamide), [0062] DMSO
(dimethyl sulfoxide) [0063] DIEA [(N,N-diisopropylethylamine)
(Hunig's base)] [0064] DMAP (4-Dimethylaminopyridine) [0065] DBU
(1,8-Diazabicyclo[5.4.0]undec-7-ene) [0066] DMPU
[1,3-Dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone] [0067] LiHMDS
(lithium hexamethyldisilazine) [0068] TMS (tetramethylsilane)
[0069] TFA (trifluoracetic acid) [0070] TLC (thin layer
chromatography) [0071] LCMS (liquid chromatographic mass
spectroscopy) [0072] MS (mass spectroscopy) giving molecular ion
[0073] NMR (nuclear magnetic resonance) [0074] The chemical shifts
values are expressed in ppm related to tetramethylsilane as
internal standard. [0075] br (broad). [0076] apt (apparent), [0077]
s (singlet), [0078] d (doublet), [0079] t (triplet), [0080] q
(quartet), [0081] dd (doublet of doublets), [0082] m (multiplet).
[0083] HPLC (high performance liquid chromatography) [0084] Mp/mp
(melting point) [0085] aq (aqueous)
[0086] All temperatures are in degrees Celsius (.degree. C.) unless
otherwise noted.
[0087] Abbreviations and acronyms used in biological screens
include: [0088] DMEM (Dulbecco's Modified Eagle's Mediunn) [0089]
FCS (fetal calf serum) [0090] PBS (phosphate buffered saline)
[0091] BCA--bicinchoninic acid
Example 1
Synthesis of 2,15-Dicycloproryl-hexadecanedioic acid
##STR00017##
[0093] Step 1--To a cooled (-78.degree. C.) sealed tube was added
isobutylene (800 mL), cyclopropane carboxylic acid (18.4 mL, 0.23
mol), t-butanol (2.0 mL) and catalytic amount of sulphuric acid
(1.0 mL). The cooled bath was removed and the reaction mixture
stirred at room temperature over a period of 72 hours. The sealed
tube was re-cooled (-78.degree. C.), opened to atmosphere and
excess isobutylene evaporated under a stream of nitrogen while
allowing the reaction mixture to attain room temperature. The
resulting mixture was diluted with ether (500 mL), washed with
saturated NaHCO.sub.3 solution (300 mL.times.3), water (300 mL),
brine (300 mL), dried over anhydrous sodium sulphate, and
evaporated under reduced pressure to give a pale, yellow liquid
which was purified by fractional distillation under reduced
pressure (120.degree. C., 30 mm Hg) to give product as a colorless
liquid 20.0 g (60%).
[0094] .sup.1H NMR (CDCl.sub.3, .delta. (ppm): 0.78-0.81 (m, 2H),
0.89-0.94 (m, 2H), 1.42 (s, 9H), 1.45-1.54 (m, 1H).
[0095] Step 2--A solution of n-butyl lithium in hexane (1.4M, 107
mL, 0.15 mol) was added dropwise to a solution of freshly distilled
diisopropylamine (19.6 mL, 0.14 mol) in dry THF (160 mL) at
-60.degree. C. under a nitrogen atmosphere. The reaction mixture
was slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -60.degree. C. and
cyclopropanecarboxylic acid tert-butyl ester (20 g, 0.14 mol) was
added dropwise over a period of 15 min. The reaction mixture was
slowly warmed to -20.degree. C. and stirred for 30 min. To the
re-cooled (-60.degree. C.) reaction mixture was added drop wise a
solution of 1,12-dibromododecane (15.15 g, 0.04 mol) in dry THF (20
mL) and DMPU (2.8 mL, 0.02 mol). Then, the temperature was allowed
to reach room temperature and was stirred at the same temperature
over a period of 16 h. The resulting reaction mixture was quenched
with saturated NH.sub.4Cl (150 mL) at 0.degree. C. and extracted
with ether (200 mL.times.3). The organic phases were combined,
dried over anhydrous Na.sub.2SO.sub.4 and the volatiles were
evaporated under reduced pressure to obtain a pale, yellow liquid
which was filtered through a silica gel (230-400) column (1.5%
ethyl acetate in pet ether) to yield 15.0 g (crude) colorless oil.
The compound was taken to the next step without further
purification or characterization.
[0096] Step 3--Formic acid (75 mL, 5 vol) was added dropwise with
vigorous stirring to the compound 3 (15.0 g, 0.03 mol) at 0.degree.
C. Then ice bath was removed and the reaction mixture was stirred
at room temperature over a period of 16 hours. The resulting
reaction mixture was diluted with toluene (50 mL) and evaporated
azeotropically to remove formic acid. The azeotropic process was
repeated three times. The residue obtained upon azeotropic
evaporation was washed with diisopropylether (50 mL.times.3) to
give a product as a white solid (3.5 g). The product was further
purified by hot ethyl acetate wash to afford a white solid (2.5 g)
(20%) having a mp of 135-140.degree. C.
[0097] .sup.1H NMR (DMSO-d6; .delta. (ppm): 0.67 (bs, 4H),
0.99-1.02 (m, 4H), 1.23 (bs, 16H), 1.40-1.43 (m, 8H), 12.00 (bs,
2H); .sup.13C NMR (DMSO-d6, .delta. ppm): 15.11, 23.20, 27.75,
29.52, 29.74, 33.85 and 176.69.
Example 2
Preparation of 2,15-dicyclopropyl-hexadecanedioic acid
di-tert-butyl ester
##STR00018##
[0099] The compound was prepared following synthetic procedure
described in Example 1.
Example 3
Preparation of
{1-[10-(1-Ethoxycarbonylmethyl-cyclopropyl)-decyl]-cyclopropyl}-acetic
acid ethyl ester
##STR00019##
[0101] Step 1--A solution of n-butyl lithium in hexane (1.4M, 426
mL, 0.59 mol) was added dropwise to a solution of freshly distilled
diisopropylamine (78.7 mL, 0.55 mol) in dry THF (560 mL) at
-60.degree. C. under a nitrogen atmosphere. The reaction mixture
was slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -60.degree. C. and
cyclopropanecarboxylic acid tert-butyl ester (80 g, 0.55 mol) was
added dropwise over a period of 30 min. The reaction mixture was
slowly warmed to -20.degree. C. and stirred for 30 min. To the
re-cooled (-60.degree. C.) reaction mixture was added drop wise
1,10-dibromododecane (42.1 g, 0.18 mol) in dry THF (56 mL) and DMPU
(11.2 mL, 0.09 mol). Then, the temperature was allowed to reach
room temperature and stirred at room temperature over a period of
16 hours. The resulting reaction mixture was quenched with
saturated NH.sub.4Cl (1.0 L) at 0.degree. C. and extracted with
ether (600 mL.times.3). The organic phases were combined, dried
over anhydrous Na.sub.2SO.sub.4 and volatiles were removed under
reduced pressure to yield a pale, yellow liquid which was filtered
through a silica gel (230-400) column (1.5% ethyl acetate in pet
ether) to yield 60.0 g crude (25%) colorless oil. The compound was
taken to the next step without further purification or
characterization.
[0102] Step 2--Formic acid (300 mL, 5 vol) was added dropwise with
vigorous stirring to compound 3 (60.0 g, 0.14 mol) at 0.degree. C.
Then, the ice bath was removed and the reaction mixture was stirred
at room temperature over a period of 16 hours. The resulting
reaction mixture was diluted with toluene (200 mL) and evaporated
azeotropically to remove formic acid. The azeotropic process was
repeated three times. The residue obtained after azeotropic solvent
removal was washed with diisopropylether (200 mL.times.3) to give
product as white solid 19 g (43.%).
[0103] .sup.1H NMR (DMSO-d6, .delta. ppm): 0.69 (bs, 4H), 0.99-1.02
(m, 4H), 1.23 (bs, 12H), 1.40-1.43 (m, 8H), 12.00 (bs, 2H).
[0104] Step 3--To a suspension of acid 4 (3.0 g, 0.01 mol) in
dichloromethane (30 mL) was added oxalyl chloride (2.7 mL, 0.02
mol) and a catalytic amount of DMF (0.06 mL) at ice temperature.
Then, the ice bath was removed and the reaction mixture was stirred
at room temperature over a period of 1 hour. The crude product
obtained upon evaporation of the volatiles was diluted with diethyl
ether (20 mL), cooled to 0.degree. C. and added to a solution of
diazomethane (4.0 g, 0.09 mol) in ether (200 mL) at the same
temperature. Then, the mixture was allowed to reach room
temperature and was stirred at room temperature over a period of 16
hours. Excess of diazomethane was removed with a stream of
nitrogen. The crude product obtained upon evaporation of the ether
was diluted with ethyl acetate (200 mL), washed with water (200 mL)
and brine (200 mL), and dried over sodium sulphate. The crude
product obtained upon evaporation of ethyl acetate was purified by
flash column chromatography to give product as a yellow solid (2.3
g) (67%).
[0105] .sup.1H NMR (DMSO-d6, .delta. (ppm): 0.70 (bs, 4H),
1.00-1.03 (m, 4H), 1.24-1.34 (m, 16H), 1.46-1.51 (m, 4H), 6.17 (s,
2H).
[0106] Step 4--To a solution of diazo compound 5 (7.0 g, 19.5 mmol)
in ethanol (50 mL) was added a freshly prepared solution of silver
benzoate (3.5 g, 15.6 mmol) in triethylamine (10.0 mL) drop wise at
reflux temperature. Then, the reaction mixture was refluxed over a
period of 6 hours, allowed to cool room temperature and
filtered.
[0107] The crude product obtained upon evaporation of the ether was
diluted with ethyl acetate (500 mL), washed with 10% sodium
bicarbonate (100 mL.times.2), water (100 mL), and brine (100 mL)
and dried over sodium sulphate. The crude product obtained upon
evaporation of ethyl acetate was purified by flash column
chromatography to give a product as a pale, yellow oil (1.51 g)
(19%).
[0108] .sup.1H NMR (DMSO-d6, .delta. ppm): 0.34-0.38 (m, 4H),
0.42-0.45 (m, 4H), 1.22-1.33 (m, 26H), 2.34 (s, 4H), 4.14 (q, J=7.2
Hz, 4H).
Example 4
Synthesis of
{1-[10-(1-carboxymethyl-cyclopropyl)-decyl]-cyclopropyl}-acetic
acid
##STR00020##
[0110] To a solution of KOH (2.7 g, 49.3 mmol) in 90% ethanol (50
mL) was added ester 6 (1.16 g, 2.90 mmol) at 80.degree. C. and
stirred at the same temperature over a period of 8.5 hours. The
crude product obtained upon evaporation of the solvent was diluted
with water (10 mL), acidified to pH=2 (1N HCl) and extracted with
ethyl acetate (100 mL.times.2). The organic layer was dried over
sodium sulphate and concentrated. The residue obtained was washed
with n-hexane (50 mL.times.2) and dried under high vacuum to give
product as an off-white solid (890 mg) (89%).
[0111] .sup.1H NMR (DMSO-d6, .delta. ppm): 0.28 (bs, 4H), 0.36 (bs,
4H), 1.23-1.28 (m, 21H), 2.15 (s, 4H), 11.96 (bs, 2H). 13C NMR
(DMSO-d6, .delta. ppm): 12.02, 17.50, 26.54, 29.51, 29.56, 29.72,
36.53 and 173.80
Example 5
Synthesis of 2,2,15,15-Dicyclopentyl-hexadecanedioic acid
##STR00021##
[0113] Cyclopentanecarboxylic acid (2.6 mL, 24.3 mmol) was added to
a stirred solution of LDA (5.33 eq, generated by adding n-butyl
lithium in hexane (1.6M, 35.0 mL, 48.7 mmol) to a solution of
diisopropylamine (7.0 mL, 48.7 mmol) in THF at -78.degree. C. under
a nitrogen atmosphere) at -30.degree. C. The reaction mixture was
slowly warmed to room temperature and stirred for 3 hours. The
reaction mixture was recooled to -10.degree. C., followed by the
rapid addition of 1,12-dibromododecane (3.0 g, 9.14 mmol) in THF
(3.0 mL). The reaction mixture was slowly warmed to room
temperature and stirred over a period of 16 hours.
[0114] The reaction mixture was quenched with a 12% HCl solution at
0.degree. C. and extracted with benzene (2.times.25 mL). The
organic layer was washed with water and brine solution, then dried
over anhydrous Na.sub.2SO.sub.4 and evaporated under reduced
pressure to obtain a crude product. The crude was purified by
titrating with hexane at -70.degree. C., filtered, and dried under
vacuum to obtain product as an off-white solid (1.7 g) (47.22%)
having amp of 117-118.degree. C.
[0115] .sup.1H NMR (DMSO-d6, .delta. (ppm): 1.22 (bs, 20H),
1.32-1.40 (m, 4H), 1.49-1.53 (bs, 12H), 1.95-2.03 (m, 4H), 11.99
(bs, 2H).
Example 6
Synthesis of 2,11-Dicyclopropyl-dodecanedioic acid
##STR00022##
[0117] Step 1--A solution of n-butyl lithium in hexane (1.4M, 16.9
mL, 0.09 mol) was added drop wise to a solution of freshly
distilled diisopropylamine (3.1 mL, 0.02 mol) in dry THF (20 mL) at
-60.degree. C. under a nitrogen atmosphere. The reaction mixture
was slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -60.degree. C. and
cyclopropanecarboxylic acid tert-butyl ester (3.1 g, 0.02 mol) was
added drop wise over a period of 10 min. The reaction mixture was
slowly warmed to -20.degree. C. and stirred for 30 min. To the
re-cooled (-60.degree. C.) above reaction mixture was added drop
wise 1,8-dibromoctane (2.0 g, 7.0 mmol) in dry THF (5 mL) and DMPU
(0.46 mL, 3.2 mmol). Then, the temperature was allowed to reach
room temperature and stirred at the same temperature over a period
of 16 hours. The resulting reaction mixture was quenched with
saturated NH.sub.4Cl (50 mL) at 0.degree. C. and the reaction
mixture extracted with ether (50 mL.times.3). The organic phases
were combined, dried over anhydrous Na.sub.2SO.sub.4 and volatiles
evaporated under reduced pressure to yield a pale, yellow liquid
which was filtered through a silica gel (230-400) column (1.5%
ethyl acetate in pet ether) to yield 1.96 g of a crude (70%)
colorless oil. The compound was taken to the next step without
further purification or characterization.
[0118] Step 2--Formic acid (6.0 mL, 10 vol) was added drop wise
with vigorous stirring to the compound 3 (600 mg, 0.14 mol) at
0.degree. C. Then, the ice bath was removed and the reaction
mixture was stirred at room temperature over a period of 16 hours.
The resulting reaction mixture was diluted with toluene (20 mL),
followed by azeotropic distillation to remove formic acid. The
azeotropic process was repeated three times. The residue obtained
upon azeotropic evaporation was washed with diisopropylether (20
mL.times.3) to give a product as a white solid (110 mg) (26%).
[0119] .sup.1H NMR (DMSO-d6, .delta. (ppm): 0.63-0.67 (m, 4H),
0.98-1.02 (m, 4H), 1.22 (bs, 8H), 1.40-1.43 (m, 8H), 12.00 (bs,
2H).
Example 7
Synthesis of
1-[8-(1-carboxymethyl-cyclopropyl)-octyl]-cyclopropyl}-acetic acid
diethyl ester
##STR00023##
[0121] Step 1--To a suspension of product 4 of Example 6 (1.0 g,
3.54 mmol) in dichloromethane (10 mL) was added oxalyl chloride
(1.0 mL, 10.6 mmol) and one drop of DMF at ice temperature. The
reaction mixture was stirred at room temperature over a period of 1
hour. The crude product obtained upon evaporation of the volatiles
was diluted with diethyl ether (20 mL) and cooled to 0.degree. C.
To the crude product was added a solution of diazomethane in ether
(40 mL) at the same temperature. Then, the mixture was stirred at
room temperature over a period of 12 hours. Excess diazomethane was
removed with a stream of nitrogen. The crude product obtained upon
evaporation of the ether was diluted with ethyl acetate (200 mL),
washed with water (50 mL) and brine (50 mL), then dried over sodium
sulphate. The crude product obtained upon evaporation of ethyl
acetate was purified by flash column chromatography to give a
product as a yellow solid (0.8 g) (68%).
[0122] .sup.1H NMR (DMSO-d6, .delta. ppm): 0.69-0.73 (m, 4H),
0.99-1.03 (m, 4H), 1.17-1.13 (m, 12H), 1.40-1.51 (m, 4H), 6.17 (s,
2H).
[0123] Step 2: To a solution of diazo compound from Step 1 (0.8 g,
2.4 mmol) in ethanol (20 mL) was added a solution of silver
triflate (300 mg, 1.35 mmol) in triethyl amine (1.5 mL) in a drop
wise fashion at reflux temperature. The reaction mixture was
refluxed for 6 hours, allowed to cool to room temperature, and
filtered. The crude product obtained upon evaporation of the
ethanol was diluted with ethyl acetate (200 mL), washed with 10%
sodium bicarbonate (50 mL) and water (50 mL), and dried over sodium
sulphate. The crude product obtained upon evaporation of ethyl
acetate was purified by flash column chromatography to give a
product as a yellow oil (0.2 g) (22%).
[0124] .sup.1H NMR (DMSO-d6, .delta. ppm): 0.34-0.38 (m, 4H),
0.41-1.45 (m, 4H), 1.15-1.14 (m, 22H), 2.23 (s, 4H), 4.11-4.18 (m,
2H).
Example 8
Synthesis of
{1-[8-(1-carboxymethyl-cyclopropyl)-octyl]-cyclopropyl}-acetic
acid
##STR00024##
[0126] To a solution of KOH (0.5 g, 9.16 mmol) in 90% ethanol (10
mL) was added the product of Example 7 (0.2 g, 0.54 mmol) at
80.degree. C. and stirred at same temperature over a period of 8.5
hours. The crude product obtained upon evaporation of the solvent
was diluted with water (5 mL), acidified to pH=2 (1N HCl),
extracted with ethyl acetate (500 mL.times.2), dried over sodium
sulphate, and concentrated. The residue obtained was washed with
n-hexane (10 mL.times.2) and dried under high vacuum to give a
product as an off-white solid (115 mg) (68%) having a mp of
100.6-101.3.degree. C.
[0127] .sup.1H NMR (DMSO-d6, .delta. (ppm): 0.26-0.29 (m, 4H),
0.34-0.37 (m, 4H), 1.22-1.27 (m, 16H), 2.12 (m, 4H), 11.95 (bs,
2H).
Example 9
Synthesis of (2,15-Dicyclopropyl-hexadecanedioic acid monoethyl
ester
##STR00025##
[0129] To a suspension of product of Example 1 (2.0 g, 5.9 mmol) in
ethanol (20 mL) was added thionyl chloride (1.65 mL, 0.02 mol) in a
dropwise fashion at ice temperature. The ice bath was removed and
reaction mixture stirred at ambient temperature over a period of 16
h. The reaction mixture was filtered to remove unreacted starting
material (0.6 gm) and residue obtained upon evaporation of the
filtrate was purified by silica gel column chromatography (5% ethyl
acetate in petroleum ether) to give product as a pale yellow liquid
100 mg. The pale yellow liquid solidified as an off-white solid
upon storage; mp 36.9-38.2.degree. C.
[0130] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.72
(m, 4H), 0.99-1.04 (m, 4H), 1.15 (t, J=7.2 Hz, 3H), 1.23 (bs, 16H),
1.40-1.46 (m, 8H), 4.02 (q, J=7.2 Hz, 2H), 11.99 (bs, 1H); MS 365.0
(M-H)
Example 10
Synthesis of 2,15-Dicyclopropyl-hexadecanedioic acid diethyl
ester
##STR00026##
[0132] To a suspension of product of Example 1 (1.0 g, 2.9 mmol) in
ethanol (10 mL) was added thionyl chloride (0.86 mL, 11.8 mmol)
drop wise at ice temperature. The ice bath was removed and reaction
mixture stirred at 80.degree. C. over a period of 16 h. The crude
product obtained upon evaporation of the volatiles was diluted with
ethyl acetate (200 mL), washed with water (50 mL.times.3) and dried
over sodium sulphate. The crude product obtained upon evaporation
of the solvent was purified through silica gel (230-400) column (4%
ethyl acetate in petroleum ether) to give product as pale yellow
liquid 0.25 g (21.5%).
[0133] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.66-0.71
(m, 4H), 1.01-1.05 (m, 4H), 1.15 (t, J=7.2 Hz, 6H), 1.23 (bs, 16H),
1.36-1.49 (m, 8H), 4.02 (q, J=7.2 Hz, 2H). .sup.13C NMR (75 MHz,
DMSO-d.sub.6) .delta. (ppm): 14.50, 15.32, 23.62, 27.62, 29.39,
29.46, 29.57, 33.57, 60.32 and 174.69. MS-395.0 (M+H)
Example 11
Synthesis of
1-[12-(1-Carbamoyl-cyclopropyl)-dodecyl]-cyclopropanecarboxylic
acid ethyl ester
##STR00027##
[0135] To a solution of product of Example 9 (1.5 g, 4.09 mmol) in
dichloromethane (15 mL) was added oxalyl chloride (0.68 mL, 8.1
mmol) and catalytic amount of DMF (0.015 mL) at ice temperature.
The ice bath was removed and reaction mixture stirred at room
temperature over a period of 1 h. The crude product obtained upon
evaporation of the solvent was diluted with DCM (10 mL), cooled to
ice bath temperature and added ammoniated DCM (100 mL) drop wise
fashion. Then reaction mixture was slowly allowed to reach room
temperature and stirred for 2 h. The resulting reaction mass was
diluted with ethyl acetate (250 mL), washed with water (100
mL.times.2) and dried over sodium sulphate. The residue obtained
upon evaporation of the volatiles was purified by silica gel
(230-400) column (40% ethyl acetate in petroleum ether) to give
product as yellow oil 0.84 g (56%). The pale yellow oil solidified
as off-white solid upon storage. mp: 49.5-51.2.2.degree. C.
[0136] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.46 (bs,
2H), 0.70-0.71 (bs, 2H), 0.87 (bs, 2H), 1.02-1.03 (bs, 2H), 1.48
(t, J=6.9 Hz, 3H), 1.23 (bs, 16H), 1.30-1.44 (m, 8H), 4.01 (q,
J=6.9 Hz, 2H), 6.79 (bs, 1H, D.sub.2O exchangeable 1H), 6.99 (bs,
.sup.1H, D.sub.2O exchangeable .sup.1H). .sup.13C NMR (75 MHz,
DMSO-d.sub.6) .delta. (ppm): 13.77, 24.70, 27.79, 29.52, 29.63,
34.36 and 175.92. MS: 366 (M+H)
Example 12
Synthesis of
(1-[12-(1-Carbamoyl-cyclopropyl)-dodecyl]-cyclopropanecarboxylic
acid
##STR00028##
[0138] To a solution of KOH (0.9 g, 16.0 mmol) in 90% ethanol (25
mL) was added product of Example 11 (0.7 g, 1.9 mmol) at 80.degree.
C. and stirred at same temperature for 8.5 h. The crude product
obtained upon evaporation of the solvent was diluted with water
(2.0 mL), acidified to pH=2 (1N HCl), and extracted with ethyl
acetate (100 mL.times.2). The organic layer was dried over sodium
sulphate and concentrated. The residue obtained was washed with
n-hexane (20.0 mL.times.2) and dried under high vacuum to give
product as white solid 470 mg (73%); mp: 116.6-118.1.degree. C.
[0139] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.45-0.48
(m, 2H), 0.63-0.67 (m, 2H), 0.86-0.89 (m, 2H), 0.99-1.02 (m, 2H),
1.30 (bs, 18H), 1.32-1.46 (m, 6H), 6.78 (bs, 1H, --CONH.sub.2;
D.sub.2O exchangeable), 6.98 (bs, 1H, --CONH.sub.2, D2O
exchangeable), 11.99 (s, 1H, --COOH, exchangeable .sup.1H).
.sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 13.83, 15.11,
23.30, 24.66, 27.75, 27.84, 29.53, 29.66, 29.74, 33.85, 34.35,
175.93 and 176.69. MS-338.0 (M+H)
Example 13
Synthesis of
1-[1,2-(1-Hydroxymethyl-cyclopropyl)-dodecyl]-cyclopropanecarboxylic
acid
##STR00029##
[0141] Step 1: To a solution of product of Example 9 (0.5 g, 1.3
mmol) in dry THF (10 mL, 20 vol) was added boranedimethyl sulfide
(0.64 mL, 6.8 mmol) at ice bath temperature. After 10 min, ice bath
was removed and the reaction mixture stirred at room temperature
over a period of 16 h. The resulting reaction mixture was quenched
with saturated ammonium chloride (20 mL) at ice bath temperature
and extracted with ethyl acetate (50 mL.times.3). The combined
organic layer was washed with water (50 mL.times.3), brine (50 mL)
and dried over sodium sulphate. The residue obtained upon
evaporation of the volatiles was purified by silica gel (230-400)
column (15% ethyl acetate in petroleum ether) to give product as
yellow oil 0.3 g (62%).
[0142] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.14-0.18
(m, 2H), 0.22-0.30 (m, 2H), 0.68-0.82 (m, 2H), 1.01-1.07 (m, 2H),
1.10-1.48 (m, 27H), 3.19 (d, J=5.7 Hz, 2H), 3.96-4.05 (m, 2H), 4.37
(t, J=5.7 Hz, 1H)
[0143] Step 2: To a solution of KOH (0.4 g, 4.5 mmol) in 90%
ethanol (15 mL) was added ester of step 1 (0.3 g, 0.8 mmol) at
80.degree. C. and stirred at same temperature over a period of 8.5
h. The crude product obtained upon evaporation of the solvent was
diluted with water (3.0 mL), acidified to pH=2 (1N HCl), and
extracted with ethyl acetate (100 mL.times.2). The organic layer
was dried over sodium sulphate and concentrated. The residue
obtained was washed with n-hexane (10.0 mL.times.2) and dried under
high vacuum to give product as white solid 240 mg (88%).
mp-73.6-75.3.degree. C.
[0144] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.15-0.18
(m, 2H), 0.27-0.30 (m, 2H), 0.63-0.67 (m, 2H), 0.99-1.02 (m, 2H),
1.15-1.28 (m, 20H), 1.35-1.40 m, 4H), 3.19 (d, J=5.1 Hz, 2H), 4.37
(t, J=5.4 Hz, 1H, --OH; D2O exchangeable 1H), 11.99 (s, 1H, --COOH;
D.sub.2O exchangeable).
[0145] .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 9.88,
15.10, 22.19, 23.29, 26.61, 27.75, 29.54, 29.64, 29.76, 29.96,
33.86, 34.33, 66.05, 176.66. MS 323.0 (M-H)
Example 14
Preparation of
1-[12-(1-Methoxymethyl-cyclopropyl)-dodecyl]-cyclopropanecarboxylic
acid ethyl ester
##STR00030##
[0147] To a solution of product of Step 1, Example 13 (0.4 g, 1.1
mmol) in dry DMF (2 mL) was added 60% sodium hydride (45 mg, 1.1
mmol) at ice bath temperature under nitrogen atmosphere. Methyl
iodide (0.14 mL, 2.2 mmol) was added drop wise at ice bath
temperature. After 10 min ice bath was removed and allowed to stir
at room temperature over a period of 2 h. The resulting reaction
mixture was quenched with ice water and diluted with ethyl acetate
(200 mL). The ethyl acetate layer was washed with water (50 mL,
.times.2), brine (50 mL) and dried over sodium sulphate. The crude
product obtained upon evaporation of the solvent was purified by
chromatography (10% ethyl acetate in petroleum ether) to give
product as pale yellow oil 400 mg (96%).
[0148] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.20-0.32
(m, 4H), 0.68-0.72 (m, 2H), 1.01-1.04 (m, 2H), 1.12-1.46 (m, 27H),
3.11 (s, 2H), 3.21 (s, 3H), 3.98-4.05 (m, 2H).
Example 15
Preparation of
1-[12-(1-Methoxymethyl-cyclopropyl)-dodecyl]-cyclopropanecarboxylic
acid
##STR00031##
[0150] To a solution of KOH (0.5 g, 9.1 mmol) in 90% ethanol (20
mL) was added product of Example 14 (0.4 g, 1.0 mmol) at 80.degree.
C. and stirred at same temperature over for 8.5 h. The crude
product obtained upon evaporation of the solvent was diluted with
water (5.0 mL), acidified to pH=2 (1N HCl), and extracted with
ethyl acetate (50 mL.times.2). The organic layer was dried over
sodium sulphate and concentrated. The residue obtained was washed
with n-hexane (10.0 mL.times.2) and dried under high vacuum to give
product as white solid 350 mg (94%). mp. 36.4-37.6.degree. C.
[0151] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.22-0.32
(m, 4H), 0.63-0.66 (m, 2H), 0.99-1.02 (m, 2H), 1.23-1.27 (m, 20H),
1.40 (bs, 4H), 3.11 (s, 2H), 3.21 (s, 3H), 11.99 (bs, 1H, --COOH;
020 exchangeable). .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta.
(ppm): 10.23, 15.09, 19.89, 23.29, 26.57, 27.75, 29.52, 29.58,
29.76, 29.85, 33.87, 34.50, 58.29, 77.45, 176.65. MS 337.0
(M-H)
Example 16
Synthesis of 2,15-Cyclobutyl-hexadecanedioic acid
##STR00032##
[0153] A solution of n-butyl lithium in hexane (1.4M, 46 mL, 0.06
mol) was added drop wise to a solution of freshly distilled
diisopropylamine (10.0 mL, 0.06 mol) in dry THF (50 mL) at
-60.degree. C. under nitrogen atmosphere. The reaction mixture was
slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -60.degree. C. and cyclobutane
carboxylic acid (3.2 g, 0.03 mol) added drop wise over a period 10
min. Reaction mixture was slowly warmed to -20.degree. C. and
stirred for 30 min. To the re-cooled (-60.degree. C.) reaction
mixture was added drop wise 1,12-dibromododecane (4.0 g, 0.01 mol)
in dry THF (4 mL). Then the temperature was allowed to reach room
temperature and stirred at same temperature over a period of 16 h.
The resulting reaction mixture was quenched with saturated NH4Cl
(100 mL) at 0.degree. C. and the reaction mixture was extracted
with ethyl acetate (100 mL.times.3). The organic phases were
combined, dried over anhydrous Na2SO4 and volatiles were evaporated
under reduced pressure to yield a pale yellow liquid which was
purified through silica gel (230-400) column (8% ethyl acetate in
petroleum ether) to give product as a white solid 160 mg (3.6%).
mp: 99.8-100.3.degree. C.
[0154] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 1.11-1.14
(m, 4H), 1.23 (m, 16H), 1.64-1.66 (m, 4H), 1.69-1.84 (m, 8H),
2.22-2.30 (m, 4H), 12.03 (s, 2H, --COOH; D.sub.2O exchangeable).
.sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 15.49, 24.93,
29.44, 29.46, 29.49, 29.77, 29.98, 38.01, 47.41, 178.41. MS: 365.0
(M-H)
Example 17
Synthesis of 2,14-Dicyclopropyl-pentadecanedioic acid
##STR00033##
[0156] Step 1: A solution of n-butyl lithium in hexane (1.6M, 114
mL, 0.18 mol) was added drop wise to a solution of freshly
distilled diisopropylamine (26.4 mL, 0.17 mol) in dry THF (180 mL)
at -60.degree. C. under nitrogen atmosphere. The reaction mixture
was slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -60.degree. C. and
cyclopropanecarboxylic acid tert-butyl ester (24.4 g, 0.17 mol)
added drop wise over a period 30 min. Reaction mixture was slowly
warmed to -20.degree. C. and stirred for 30 min. To the re-cooled
(-60.degree. C.) reaction mixture was added drop wise
1,11-dibromoundecane (18.0 g, 0.05 mol) in dry THF (18 mL) and DMPU
(3.6 g, 0.02 mol). Then the temperature was allowed to reach room
temperature and stirred at same temperature over a period of 16 h.
The resulting reaction mixture was quenched with saturated NH4Cl
(500 mL) at 0.degree. C. and the reaction mixture was extracted
with ethyl acetate (300 mL.times.2). The organic phases were
combined, dried over anhydrous Na.sub.2SO.sub.4 and volatiles were
evaporated under reduced pressure to yield a pale yellow liquid
which was filtered through silica gel (230-400) column (1.5% ethyl
acetate in petroleum ether) to give 24 g (96%) crude product as
colorless oil. The product obtained was taken to next step without
characterization.
[0157] Step 2: Formic acid (240 mL, 10 vol) was added drop wise
with vigorous stirring to the compound of Step 1 (24.0 g, 0.05 mol)
at 0.degree. C. Then ice bath was removed and reaction mixture
stirred at room temperature over a period of 16 h. The resulting
reaction mixture was diluted with toluene (200 mL) and evaporated
azeotropically to remove formic acid. The azeotropic evaporation
was repeated three times. The residue obtained upon azeotropic
evaporation was washed with diisopropylether (100 mL.times.3) to
give product as white solid 5.0 g (28%). mp: 114.7-117.7.degree.
C.
[0158] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.67
(m, 4H), 0.99-1.23 (m, 4H), 1.23 (s, 14H), 1.40 (bs, 8H), 12.00
(bs, 2H, --COOH; D2O exchangeable).
[0159] .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 15.10,
23.30, 27.75, 29.51, 29.74, 33.85, 176.68. MS: 323.0 (M-H).
Example 18
Synthesis of
1-[10-(1-Carboxymethyl-cyclobutyl)-decyl]-cyclopropanecarboxylic
acid
##STR00034##
[0161] Step 1: A solution of n-butyl lithium in hexane (1.4M, 45.75
mL, 0.07 mol) was added drop wise to a solution of freshly
distilled diisopropylamine (10.2 mL, 0.06 mol) in dry THF (200 mL)
at -60.degree. C. under nitrogen atmosphere. The reaction mixture
was slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -60.degree. C. and
cyclopropanecarboxylic acid tert-butyl ester (9.48 g, 0.06 mol)
added drop wise over a period 30 min. Reaction mixture was slowly
warmed to -20.degree. C. and stirred for 30 min. To the re-cooled
(-60.degree. C.) reaction mixture was added drop wise
1,10-dibromodecane (20.0 g, 0.06 mol) in dry THF (20 mL) and DMPU
(1.79 g, 0.01 mol). Then the temperature was allowed to reach room
temperature and stirred at same temperature over a period of 16 h
The resulting reaction mixture was quenched with saturated
NH.sub.4Cl (500 mL) at 0.degree. C. and the reaction mixture was
extracted with ethyl acetate (300 mL.times.3). The organic phases
were combined, dried over anhydrous Na.sub.2SO.sub.4 and volatiles
were evaporated under reduced pressure to yield a pale yellow
liquid which was purified through silica gel (230-400) column (0.5%
ethyl acetate in petroleum ether) to give product as colorless oil
8.0 g (33%).
[0162] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.62-0.64
(m, 2H), 0.94-0.98 (m, 2H), 1.25 (bs, 12H), 1.37 (s, 9H), 1.39-1.43
(m, 4H), 1.73-1.80 (m, 2H), 3.52 (t, J=6.6 Hz, 2H).
[0163] Step 2: A solution of n-butyl lithium in hexane (1.4M, 84.2
mL, 0.11 mol) was added drop wise to a solution of freshly
distilled diisopropylamine (18.4 mL, 0.11 mol) in dry THF (100 mL)
at -78.degree. C. under nitrogen atmosphere. The reaction mixture
was slowly warmed to -30.degree. C. and stirred for 30 min. To the
reaction mixture cyclobutaric acid (5.88 g, 0.05 mol) added drop
wise over a period 30 min. Reaction mixture was slowly warmed to
room temperature and stirred for 3 h. To the re-cooled (-10.degree.
C.) reaction mixture was added drop wise product of Step 1 (8.0 g,
0.02 mol) in dry THF (6.6 mL). Then the temperature was allowed to
reach room temperature and stirred at same temperature over a
period of 16 h. The resulting reaction mixture was quenched with
saturated NH.sub.4Cl (200 mL) at 0.degree. C. and the reaction
mixture was extracted with ethyl acetate (500 mL). The organic
phases were combined, dried over anhydrous Na.sub.2SO.sub.4 and
volatiles were evaporated under reduced pressure to yield a pale
yellow liquid which was purified through silica gel (230-400)
column (1% ethyl acetate in petroleum ether) to give product as
colorless pale yellow oil 2.0 g (23%).
[0164] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.62-0.68
(m, 2H), 0.91-1.01 (m, 2H), 1.10-1.25 (m, 2H), 1.37-1.39 (bs, 13H),
1.42-1.45 (bs, 12H), 1.64-1.69 (m, 2H), 1.73-1.84 (m, 4H),
2.41-2.30 (m, 2H), 12.03 (bs, 1H).
[0165] Step 3: To a suspension of product of Step 2 (2.0 g, 5.2
mmol) in dry THF (20 mL) was added triethyl amine (0.73 mL, 5.2
mmol) at ice temperature. After 15 min ethyl chloroformate (0.5 mL,
5.2 mmol) was added and the reaction mixture was stirred at same
temperature over a period of 15 min. The resultant reaction mixture
cooled to 0.degree. C. and a solution of diazomethane (2.2 g, 0.05
mol) in ether (250 mL) was added. Then the mixture was allowed to
reach room temperature and stirred over a period of 16 h. Excess of
diazomethane was removed with steam of nitrogen, the crude product
obtained upon evaporation of the ether was diluted with ethyl
acetate (500 mL), washed with water (100 mL.times.2), brine (100
mL) and dried over sodium sulphate. The crude product obtained upon
evaporation of ethyl acetate was purified by flash column
chromatography (4% ethyl acetate in petroleum ether) to give
product as yellow liquid 0.51 g (24%).
[0166] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (Ppm): 0.55-0.63
(m, 2H), 1.08-1.12 (m, 2H), 1.26 (m, 15H), 1.43-1.48 (m, 12H),
1.64-1.69 (m, 2H), 1.72-1.91 (m, 4H), 2.34-2.38 (m, 2H), 5.21 (s,
1H).
[0167] Step 4: To a solution of diazo compound from Step 3 (0.51 g,
1.2 mmol) in ethanol (20 mL) was added a freshly prepared solution
of silver benzoate (0.22 g, 0.9 mmol) in triethyl amine (1.0 mL) in
a drop wise fashion at reflux temperature. Then the reaction
mixture was refluxed over a period of 16 h, allowed to cool room
temperature and filtered. The crude product obtained upon
evaporation of the ethanol was diluted with ethyl acetate (200 mL),
washed with 10% sodium bicarbonate (50 mL), water (50 mL), brine
(50 mL) and dried over sodium sulphate. The crude product obtained
upon evaporation of ethyl acetate was purified by flash column
chromatography (2% ethyl acetate in petroleum ether) to give
product 0.128 g (35%) as pale yellow oil.
[0168] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.62-0.68
(m, 2H), 0.97-0.99 (m, 2H), 1.14-1.39 (m, 17H), 1.43-1.46 (m, 15H),
1.70-1.90 (m, 6H), 2.37 (s, 2H), 4.03 (q, J=7.2 Hz, 2H).
[0169] Step 5: To a solution of KOH (0.2 g, 3.5 mmol) in 90%
ethanol (10 mL) was added product of Step 4 (0.18 g, 0.4 mmol) at
80.degree. C. and stirred at same temperature over a period of 8.5
h. The crude product obtained upon evaporation of the solvent was
diluted with water (2.0 mL), acidified to pH=2 (1N HCl), and
extracted with ethyl acetate (50 mL.times.2). The organic layer was
dried over sodium sulphate and concentrated. The residue obtained
150 mg (89%) was taken to next step without further purification
and characterization.
[0170] Step 6: Formic acid (1.5 mL, 10 vol) was added drop wise
with vigorous stirring to the product of Step 5 (0.15 g, 0.3 mmol)
at 0.degree. C. Then ice bath was removed and reaction mixture
stirred at room temperature over a period of 16 h. The resulting
reaction mixture was diluted with toluene (4 mL) and evaporated
azeotropically to remove formic acid. The azeotropic evaporation
was repeated three times. The residue obtained upon azeotropic
evaporation was washed with diisopropylether (5 mL.times.3) to give
product as Off-white solid 120 mg (83%). mp: 62.0.degree.
C.-63.9.degree. C.
[0171] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.67
(m, 2H), 1.00-1.09 (m, 2H), 1.23 (bs, 14H), 1.40-1.49 (m, 6H),
1.71-1.89 (m, 6H), 2.30 (s, 2H), 11.93-12.09 (bs, 2H, COOH
exchangeable .sup.1H). .sup.1H NMR (300 MHz, DMSO-d.sub.6-D.sub.2O)
.delta. (ppm): 0.63-0.67 (m, 2H), 0.99-1.01 (m, 2H), 1.23 (bs,
14H), 1.40-148 (m, 6H), 1.70-1.88 (m, 6H), 2.30 (s, 2H).
Example 19
Preparation of
1-[12-(1-Cyano-cyclopropyl)-dodecyl]-cyclopropanecarboxylic
acid
##STR00035##
[0173] Step 1: A solution of n-butyl lithium in hexane (1.4M, 95.7
mL, 0.13 mol) was added drop wise to a solution of freshly
distilled diisopropylamine (18.75 mL, 0.12 mol) in dry THF (400 mL)
at -60.degree. C. under nitrogen atmosphere. The reaction mixture
was slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -60.degree. C. and
cyclopropanecarboxylic acid tert-butyl ester (17.34 g, 0.12 mol)
added drop wise over a period 30 min. Reaction mixture was slowly
warmed to -20.degree. C. and stirred for 30 min. To the re-cooled
(-60.degree. C.) reaction mixture was added drop wise
1,12-dibromododecane (40.0 g, 0.12 mol) in dry THF (40 mL) and DMPU
(3.12 g, 0.02 mol). Then the temperature was allowed to reach room
temperature and stirred at same temperature over a period of 16 h.
The resulting reaction mixture was quenched with saturated
NH.sub.4Cl (500 mL) at 0.degree. C. and the reaction mixture was
extracted with ethyl acetate (400 mL.times.3). The organic phases
were combined, dried over anhydrous Na.sub.2SO.sub.4 and volatiles
were evaporated under reduced pressure to yield a pale yellow
liquid which was purified through silica gel (230-400) column (0.5%
ethyl acetate in petroleum ether) to give product as pale yellow
liquid 14.0 g (29%).
[0174] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 1.55-1.61
(m, 2H), 1.08-1.12 (m, 2H), 1.35 (bs, 13H), 1.49 (bs, 16H),
1.64-1.89 (m, 2H), 3.42 (t, J=6.9 Hz, 2H).
[0175] Step 2: A solution of n-butyl lithium in hexane (1.4M, 58.7
mL, 0.08 mol) was added drop wise to a solution of freshly
distilled diisopropylamine (11.8 mL, 0.07 mol) in dry THF (100 mL)
at -60.degree. C. under nitrogen atmosphere. The reaction mixture
was slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -78.degree. C. and cyclopropane
carbonitrile (5.17 g, 0.07 mol) added drop wise over a period 30
min and stirred at same temperature over a period of 1 h. To the
reaction mixture was added drop wise product of Step 1 (10.0 g,
0.02 mol) in dry THF (40 mL) and DMPU (1.6 g, 0.01 mol). After 2 h
stirring at -78.degree. C. the reaction mixture was slowly allowed
to reach room temperature and continued stirring over a period of
16 h. The resulting reaction mixture was quenched with saturated
NH.sub.4Cl (250 mL) at 0.degree. C. and the reaction mixture was
extracted with ethyl acetate (300 mL.times.2). The organic phases
were combined, dried over anhydrous Na.sub.2SO.sub.4 and volatiles
were evaporated under reduced pressure to yield a pale yellow
liquid which was purified through silica gel (230-400) column (0.5%
ethyl acetate in petroleum ether) to give product as pale yellow
liquid 2.0 g (20%). The crude product so obtained was used in next
step without further purification.
[0176] Step 3: Formic acid (20 mL, 10 vol) was added drop wise with
vigorous stirring to the product of Step 2 (2.0 g, 5.3 mmol) at
0.degree. C. Then ice bath was removed and reaction mixture stirred
at room temperature over a period of 16 h. The resulting reaction
mixture was diluted with toluene (20 mL) and evaporated
azeotropically to remove formic acid. The azeotropic evaporation
was repeated three times. The residue obtained upon azeotropic
evaporation was purified through silica gel column chromatography
(7% ethyl acetate in petroleum ether) to give product as Off-white
solid 620 mg (36%). mp: 44.4.degree. C.-46.9.degree. C.
[0177] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.64-0.68
(m, 2H), 0.84-1.86 (m, 2H), 1.00-1.02 (m, 2H), 1.12-1.16 (m, 2H),
1.24 (bs, 16H), 1.40-1.43 (m, 8H), 11.99 (bs, 1H, COOH exchangeable
.sup.1H). MS 318 (M-H)
Example 20
Synthesis of of
1-{12-[1-(1H-Tetrazol-5-yl)-cyclopropyl]-dodecyl}-cyclopropanecarboxylic
acid
##STR00036##
[0179] To a solution of Step 3 product of Example 19 (0.38 g, 1.8
mmol) in nitrobenzene (4 mL) were added sodium azide (0.46 g, 7.1
mmol) and triethyl amine hydrochloride (0.98 g, 7.1 mmol) at room
temperature. The reaction mixture was allowed to stir at
130.degree. C. over a period of 24 h. The reaction mixture was
diluted with diethyl ether (50 mL) and extracted with 2% NaOH
solution (50 mL.times.2). The combined aqueous layer was acidified
to P.sup.H=5 (1N HCl), extracted with ethyl acetate (50
mL.times.2), dried over sodium sulphate. The residue obtained upon
evaporation of the solvent was purified through silica gel
(230-400) column (50% ethyl acetate in petroleum ether) to give
product as Off-white solid 120 mg (27%). mp-133.2.degree.
C.-135.0.degree. C.
[0180] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.67
(m, 2H), 0.99-1.02 (m, 4H), 1.08-1.15 (m, 2H), 1.22 (bs, 18H), 1.40
(bs, 4H), 1.70-1.75 (m, 2H), 11.97-12.14 (bs, 1H, COOH exchangeable
.sup.1H). .sup.1H NMR (300 MHz, DMSO-d.sub.6-D.sub.2O) .delta.
(ppm): 0.64-0.68 (m, 2H), 0.99-1.06 (m, 6H), 1.16 (bs, 18),
1.30-1.36 (bs, 4H), 1.66-1.71 (m, 2H), .sup.13C NMR (75 MHz,
DMSO-d.sub.6) .delta. (ppm): 15.12, 15.43, 16, 41, 23.32, 27, 26,
27.75, 29, 40, 29.48, 29, 72, 33.83, 35.38, 160.48, and 176.71. MS:
361.5 (M-H)
Example 21
Preparation of 2,13-dicyclopropyll-tetradecanedioic acid
##STR00037##
[0182] Step 1: A solution of n-butyl lithium in hexane (1.4M, 22.8
mL, 0.32 mol) was added drop wise to a solution of freshly
distilled diisopropylamine (46.0 mL, 0.30 mol) in dry THF (300 mL)
at -60.degree. C. under nitrogen atmosphere. The reaction mixture
was slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -60.degree. C. and
cyclopropanecarboxylic acid tert-butyl ester (42.6 g, 0.30 mol)
added drop wise over a period 30 min. Reaction mixture was slowly
warmed to -20.degree. C. and stirred for 30 min. To the re-cooled
(-60.degree. C.) reaction mixture was added drop wise
1,10-dibromodecane (30.0 g, 0.10 mol) in dry THF (30 mL) and DMPU
(6.4 g, 0.05 mol). Then the temperature was allowed to reach room
temperature and stirred at same temperature over a period of 16 h.
The resulting reaction mixture was quenched with saturated
NH.sub.4Cl (500 mL) at 0.degree. C. and the reaction mixture was
extracted with ethyl acetate (300 mL.times.3). The organic phases
were combined, dried over anhydrous Na.sub.2SO.sub.4 and volatiles
were evaporated under reduced pressure to yield a pale yellow
liquid which was filtered through silica gel (230-400) column (1.5%
ethyl acetate in petroleum ether) to give 28 g (66%) crude product
as colorless oil. The product obtained was taken to next step
without characterization.
[0183] Step 2: Formic acid (140 mL, 5 vol) was added drop wise with
vigorous stirring to the product of Step 1 (28.0 g, 0.06 mol) at
0.degree. C. Then ice bath was removed and reaction mixture stirred
at room temperature over a period of 16 h. The resulting reaction
mixture was diluted with toluene (100 mL) and evaporated
azeotropically to remove formic acid. The azeotropic evaporation
was repeated three times. The residue obtained upon azeotropic
evaporation was washed with diisopropylether (100 mL.times.3) to
give product as white solid 7.0 g (34.%). mp: 142.5.degree.
C.-144.3.degree. C.
[0184] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.67
(m, 4H), 0.98-0.10 (m, 4H), 0.14-0.17 (m, 12H), 1.40 (bs, 8H),
11.98 (s, 2H). .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm):
15.09, 23.29, 27.74, 29.51, 29.74, 33.85 and 176.68. MS: 309
(M-H)
Example 22
Synthesis of 2,13-dicyclopropyll-tetradecanedioic acid monoethyl
ester
##STR00038##
[0186] To a suspension of product from Example 21 (7.0 g, 0.02 mol)
in ethanol (70 mL) was added thionyl chloride (1.65 mL, 0.02 mol)
drop wise at ice temperature. Ice bath was removed and reaction
mixture stirred at ambient temperature over a period of 16 h.
Filtered the reaction mixture to remove un reacted starting
material (2.1 g) and residue obtained upon evaporation of the
filtrate was purified by column chromatography (5% ethyl acetate in
petroleum ether) to give product as a pale yellow liquid 1.0 g
(13.1%).
[0187] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.72
(m, 4H), 0.98-1.04 (m, 4H), 1.09 (t, J=6.9 Hz, 3H), 1.23 (bs, 12H),
1.40-1.48 (m, 8H), 4.02 (q, J=6.9 Hz, 2H), 11.99 (bs, 1H). MS: 337
(M-H)
Example 23
Synthesis of
1-[10-(1-Carboxymethyl-cyclopropyl)-decyl]-cyclopropanecarboxylic
acid
##STR00039##
[0189] Step 1: To a suspension of acid product of Example 22 (1.0
g, 2.9 mmol) in dichloromethane (15 mL) was added oxalyl chloride
(0.37 mL, 4.3 mmol) and catalytic amount of DMF (0.01 mL) at ice
temperature. Then ice bath was removed and reaction mixture was
stirred at room temperature over a period of 1 h. The crude product
obtained upon evaporation of the volatiles was diluted with diethyl
ether (5.0 mL), cooled to 0.degree. C. and a solution of
diazomethane (1.24 g, 0.03 mol) in ether (100 mL) added. Then the
mixture was allowed to reach room temperature and stirred over a
period of 16 h. Excess of diazomethane was removed with steam of
nitrogen, the crude product obtained upon evaporation of the ether
was diluted with ethyl acetate (200 mL), washed with water (100
mL), brine (100 mL), dried over sodium sulphate. The crude product
obtained upon evaporation of ethyl acetate was purified by flash
column chromatography to give product as a yellow liquid 0.7 g
(65%).
[0190] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.69-0.72
(m, 4H), 1.00-1.05 (m, 4H), 1.15 (t, J=7.2 Hz, 3H), 1.23 (bs, 12H),
1.32-1.35 (m, 4H), 1.44-1.51 (m, 4H), 4.02 (q, J=7.2 Hz, 2H), 6.17
(s, 1H). LC-MS: 363.5 (M+H)
[0191] Step 2: To a solution of diazo compound of Step 1 (0.7 g,
1.9 mmol) in ethanol (20 mL) was added a freshly prepared solution
of silver benzoate (0.35 g, 1.5 mmol) in triethyl amine (3.0 mL) in
a drop wise fashion at reflux temperature. Then the reaction
mixture was refluxed over a period of 6 h, allowed to cool room
temperature and filtered. The crude product obtained upon
evaporation of the ether was diluted with ethyl acetate (200 mL),
washed with 10% sodium bicarbonate (50 mL), water (50 mL), brine
(50 mL) and dried over sodium sulphate. The crude product obtained
upon evaporation of ethyl acetate was purified by flash column
chromatography (4% ethyl acetate in petroleum ether) to give
product 0.35 g (47.9%) as pale yellow oil.
[0192] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.27-0.30
(m, 2H), 0.36-0.39 (m, 2H), 0.69-0.72 (m, 2H), 1.01-1.05 (m, 2H),
1.12-1.20 (m, 6H), 1.23-1.26 (m, 16H), 1.36-1.47 (m, 4H), 2.20 (s,
2H), 3.98-4.08 (m, 4H). MS: 381.7 (M+H)
[0193] Step 3: To a solution of KOH (0.86 g, 15.4 mmol) in 90%
ethanol (10 mL) was added product of Step 2 (0.35 g, 0.91 mmol) at
80.degree. C. and stirred at same temperature over a period of 8.5
h. The crude product obtained upon evaporation of the solvent was
diluted with water (2.0 mL), acidified to pH=2 (1N HCl), and
extracted with ethyl acetate (50 mL.times.2), The organic layer was
dried over sodium sulphate and concentrated. The residue obtained
was washed with n-hexane (10.0 mL.times.2) and dried under high
vacuum to give product as a white solid 250 mg (83%). mp:
71.9.degree. C.-74.3.degree. C.
[0194] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.26-0.29
(m, 2H), 0.34-0.37 (m, 2H), 0.63-0.66 (m, 2H), 0.99-1.02 (m, 2H),
1.22 (bs, 12H), 1.27 (bs, 4H), 1.40 (bs, 4H), 2.12 (s, 2H), 11.97
(bs, 2H). .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 12.01,
15.10, 17.50, 23.30, 26.53, 27.74, 29.50, 29.55, 29.72, 29.74,
33.85, 36.52, 173.80 and 176.68. MS: 323.5 (M-H)
Example 24
Synthesis of 1-(13-Carboxy-13-methyl-tetradecyl)cyclopropane
carboxylic acid
##STR00040##
[0196] Step 1: A solution of n-butyl lithium in hexane (1.4M, 41.8
mL, 0.06 mol) was added drop wise to a solution of freshly
distilled diisopropylamine (9.3 mL, 0.06 mol) in dry THF (200 mL)
at -60.degree. C. under nitrogen atmosphere. The reaction mixture
was slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re cooled to -60.degree. C. and
cyclopropanecarboxylic acid tert-butyl ester (8.67 g, 0.06 mol)
added drop wise over a period 30 min. Reaction mixture was slowly
warmed to -20.degree. C. and stirred for 30 min. To the re-cooled
(-60.degree. C.) reaction mixture was added drop wise
1,12-dibromododecane (20.0 g, 0.06 mol) in dry THF (20 mL) and DMPU
(1.56 g, 0.01 mol). Then the temperature was allowed to reach room
temperature and stirred at same temperature over a period of 16 h.
The resulting reaction mixture was quenched with saturated
NH.sub.4Cl (500 mL) at 0.degree. C. and extracted with ethyl
acetate (300 mL.times.3). The organic phases were combined, dried
over anhydrous Na.sub.2SO.sub.4 and volatiles were evaporated under
reduced pressure to yield a pale yellow liquid which was filtered
through silica gel (230-400) column (0.5% ethyl acetate in
petroleum ether) to give 2.5 g (10.6%) product as a yellow color
viscous oil.
[0197] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.66
(m, 2H), 0.94-0.98 (m, 2H), 1.24 (bs, 17H), 1.33-1.39 (m, 12H),
1.73-1.83 (m, 2H), 3.33-3.54 (m, 2H).
[0198] Step 2: A solution of n-butyl lithium in hexane (1.4M, 19.42
mL, 0.02 mol) was added drop wise to a solution of freshly
distilled diisopropylamine (4.19 mL, 0.02 mol) in dry THF (30 mL)
at -60.degree. C. under nitrogen atmosphere. The reaction mixture
was slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -60.degree. C. and isobutyric
acid (0.05 mL, 0.01 mol) added drop wise over a period 30 min.
Reaction mixture was slowly warmed to -20.degree. C. and stirred
for 30 min. To the re-cooled (-60.degree. C.) reaction mixture was
added drop wise bromo intermediate of Step 1 (2.0 g, 0.005 mol) in
dry THF (5.0 mL). Then the temperature was allowed to reach room
temperature and stirred at same temperature over a period of 16 h.
The resulting reaction mixture was quenched with saturated
NH.sub.4Cl (300 mL) at 0.degree. C. and extracted with ethyl
acetate (100 mL.times.3). The organic phases were combined, dried
over anhydrous Na.sub.2SO.sub.4 and volatiles were evaporated under
reduced pressure to give pale yellow liquid which was filtered
through silica gel (230-400) column (2.0% ethyl acetate in
petroleum ether) to give 1.3 g crude (64%) colorless oil. The
compound was taken to next step without further purification and
characterization.
[0199] Step 3: Formic acid (6.5 mL, 5 vol) was added drop wise with
vigorous stirring to the compound 4 (1.3 g, 0.003 mol) at 0.degree.
C. Then ice bath was removed and reaction mixture stirred at room
temperature over a period of 16 h. The resulting reaction mixture
was diluted with toluene (10 mL) and evaporated azeotropically to
remove formic acid. The azeotropic evaporation was repeated two
times. The residue obtained upon azeotropic evaporation was washed
with 10% dichlorormethane in hexane (20 mL.times.3) to give product
as an off-solid 350 mg (31%). mp: 85.6-86.7.degree. C.
[0200] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.66
(m, 2H), 0.99-1.02 (m, 2H), 1.06 (s, 6H), 1.23 (bs, 18H), 1.40-1.44
(m, 6H), 12.00 (bs, 2H). .sup.13C NMR (75 MHz, DMSO-d.sub.6)
.delta. (ppm): 15.09, 23.31, 24.94, 25.47, 27.75, 29.50, 29.74,
30.04, 33.86, 41.66, 176.68 and 179.27. MS-339.0 (M-H)
Example 25
Synthesis of
{1-[12-(1-Hydroxymethyl-cyclopropyl)-dodecyl]-cyclopropyl}-methanol
##STR00041##
[0202] To a solution of product of Example 1 (2.0 g, 5.9 mmol) in
dry THF (20 mL, 10 vol) was added boranedimethyl sulfide (5.61 mL,
59.1 mmol) at ice bath temperature. After 10 min ice bath was
removed and the reaction mixture was stirred at room temperature
for 5 h. The resulting reaction mixture was quenched with saturated
ammonium chloride (100 mL) at ice bath temperature and extracted
with ethyl acetate (100 mL.times.3). The combined organic layer was
washed with water (100 mL), brine (100 mL) and dried over sodium
sulphate. The residue obtained upon evaporation of the volatiles
was washed with ratio (0.5:9.5) of dichloromethane: petroleum ether
to give product as an off-white solid 1.5 g (81.9%). mp:
63.2-64.6.degree. C.
[0203] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.15-0.18
(m, 4H), 0.27-0.30 (m, 4H), 1.23 (bs, 16H), 1.28 (bs, 8H), 3.19 (d,
J=5.7 Hz, 4H), 4.37 (t, J=5.7 Hz, 2H, --OH, Exchangeable proton).
.sup.1H NMR (300 MHz, DMSO-d.sub.6-D2O) .delta. (ppm): 0.13-0.16
(m, 4H), 0.24-0.28 (m, 4H), 1.20 (bs, 16H), 1.25 (bs, 8H), 3.16 (s,
4H). .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 9.86,
22.16, 26.63, 29.60, 29.68, 30.00, 34.37 and 66.04.
Example 26
Synthesis of 1,16-Dimethoxy-2,15-dicyclopropyl-hexadecane
##STR00042##
[0205] To a solution of product of Example 25 (1.0 g, 3.2 mmol) in
dry DMF was added 60% sodium hydride (0.38 g, 9.6 mmol) at ice bath
temperature in three portions in intervals of 5 min under nitrogen
atmosphere. Methyl iodide (1.19 mL, 19.3 mmol) added drop wise at
ice bath temperature. After 10 min ice bath was removed and allowed
to stir at room temperature over a period of 2 h. The resulting
reaction mixture was quenched with ice water and diluted with ethyl
acetate (300 mL). The ethyl acetate layer was washed with water
(100 mL.times.2) and dried over sodium sulphate. The crude product
obtained upon evaporation of the solvent was purified by
chromatography (3% ethyl acetate in petroleum ether) to give
product as pale yellow oil 600 mg (55%).
[0206] .sup.1H NMR (300 MHz, CDCl3) .delta. (ppm): 0.31-0.39 (m,
8H), 1.27 (bs, 18H), 1.35 (bs, 8H), 3.19 (s, 4H), 3.35 (s, 6H).
.sup.13C NMR (75 MHz, CDCl3) .delta. (ppm): 10.05, 19.76, 26.59,
29.67, 29.72, 29.97, 34.39, 58.62 and 78.25.
Example 27
Synthesis of 1,1'-undecane-1,11-diyldicyclopropanecarbonitrile
##STR00043##
[0208] A solution of n-butyl lithium in hexane (1.8M, 297 mL, 0.53
mol) was added drop wise to a solution of freshly distilled
diisopropylamine (78.3 mL, 0.50 mol) in dry THF (400 mL) at
-60.degree. C. under nitrogen atmosphere. The reaction mixture was
slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -78.degree. C., DMPU (8.16 g,
0.06 mol) was added drop wise and stirred for 15 min. Then
cyclopropane carbonitrile (34.17 g, 0.50 mol) was added drop wise
over a period 30 min and stirred for 1 h. To the reaction mixture
was added drop wise 1,11-dibromoundecane (40.0 g, 0.12 mol) in dry
THF (40 mL) and continued stirring at -78.degree. C. for 2 h. The
reaction mixture was slowly allowed to reach room temperature and
continued stirring over a period of 16 h. The resulting reaction
mixture was quenched with saturated NH.sub.4Cl (1.0 L) at 0.degree.
C. and the reaction mixture was extracted with ethyl acetate (600
mL.times.2). The organic phases were combined, dried over anhydrous
Na.sub.2SO.sub.4 and volatiles were evaporated under reduced
pressure to yield a pale yellow liquid which was purified through
silica gel (230-400) column (5% ethyl acetate in petroleum ether)
to give product as a pale yellow liquid 25 g (68.5%).
[0209] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.84-0.85
(m, 4H), 1.07-1.14 (m, 4H), 1.23-1.25 (bs, 14H), 1.42 (bs, 8H). MS:
287 (M+H), 304 (M+NH.sub.4)
Example 28
Synthesis of 2,14-Dicyclopropyl-pentadecanedioic acid monoethyl
ester
##STR00044##
[0211] Step 1: To a solution of product of Example 27 (15.0 g,
0.052 mol) in ethanol (150 mL) was added water (75 mL), potassium
hydroxide (88 g, 1.57 mol) and the reaction mass was stirred at
140.degree. C. over a period of 72 h. The crude product obtained
upon evaporation of the solvent was diluted with water (150.0 mL),
acidified to pH=2 (6N HCl) to obtain pale yellow precipitate. The
solid obtained upon filtration was washed with diisopropyl ether
(50 mL.times.2) and dried to give product as pale yellow solid 15 g
(88.33%); identical to product of Example 17.
[0212] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.70
(m, 4H), 0.92-0.98 (m, 4H), 1.21 (s, 14H), 1.38 (bs, 8H), 11.96
(bs, 2H). MS: 323 (M-H)
[0213] Step 2: To a suspension of product of Step 1 (15.0 g, 0.04
mol) in ethanol (150 mL) was added thionyl chloride (3.38 mL, 0.04
mol) drop wise at ice temperature. Ice bath was removed and
reaction mixture stirred at 25.degree. C. over a period of 16 h.
The crude product obtained upon evaporation of the volatiles was
purified through silica gel (230-400) column (5% ethyl acetate in
petroleum ether) to give product as an orange liquid 4.0 g (24.5%)
and recovered starting material (6.0 g, 40%).
[0214] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.73
(m, 4H), 0.99-1.02 (m, 4H), 1.13 (t, J=6.9 Hz, 3H), 1.22 (bs, 14H),
1.38-1.42 (m, 8H), 4.01 (q, J=6.9 Hz, 2H), 11.98 (bs, 1H, --COOH
Exchangeable 1H). MS: 351 (M-H)
Example 29
1-[11-(1-Hydroxymethyl-cyclopropyl)-undecyl]-cyclopropane
carboxylic acid ethyl ester
##STR00045##
[0216] To a solution of product of Example 28 (4.0 g, 1.3 mol) in
dry THF (40 mL) was added boranedimethyl sulfide (5.3 mL, 0.056
mol) at ice bath temperature. After 10 min, ice bath was removed
and the reaction mixture stirred at room temperature over a period
of 5 h. The resulting reaction mixture was quenched with saturated
ammonium chloride (100 mL) at ice bath temperature and extracted
with ethyl acetate (200 mL.times.3). The combined organic layer was
washed with water (100 mL), brine (100 mL) and dried over sodium
sulphate. The residue obtained upon evaporation of the volatiles
was purified by silica gel (230-400) column (10% ethyl acetate in
petroleum ether) to give product as orange oil 2.7 g (70%).
[0217] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.14-0.17
(m, 2H), 0.26-0.29 (m, 2H), 0.68-0.71 (m, 2H), 1.00-1.04 (m, 2H),
1.14 (t, J=7.2 Hz, 3H), 1.22-1.27 (m, 18H), 1.34-1.38 (m, 4H), 3.18
(d, J=5.7 Hz, 2H), 4.01 (q, J=7.2 Hz, 2H), 4.36 (t, J=5.7 Hz, 1H),
MS: 321 (M-H.sub.2O).
Example 30
Synthesis of
1-[11-(1-Carboxymethyl-cyclopropyl)-undecyl]-cyclopropanecarboxylic
acid
##STR00046##
[0219] Step 1: A solution of dichloromethane (15 mL) and oxalyl
chloride (1.5 mL, 0.017 mol) in a 50 mL flask was cooled in a dry
ice-acetone bath (-78.degree. C.). A solution of dimethyl sulfoxide
(2.5 mL, 0.035 mol) dissolved in dichloromethane (7 mL) was added
to the stirred oxalyl chloride solution at -70.degree. C. The
reaction mixture was stirred for 5 minutes, and a solution of
product of Example 29 (2.7 g, 7.9 mmol) in dichloromethane (5 mL)
was added drop wise over a period of 5 minutes. Stirring was
continued for an additional 45 min at -70.degree. C. and
triethylamine (7.3 mL, 0.08 mol) was added. The reaction mixture
was stirred for 5 min and then allowed to warn to room temperature.
After 1 hour, water (200 mL) was added and the aqueous layer was
extracted with additional dichloromethane (500 mL). The organic
layer was combined, washed with brine (200 mL) and dried over
anhydrous sodium sulphate. The crude product obtained upon
evaporation of the solvent was purified by silica gel column (2.0%
ethyl acetate in petroleum ether) to give product as a yellow
liquid 2.2 g (82%).
[0220] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.69-0.71
(m, 2H), 0.92-0.94 (m, 2H), 1.00-1.03 (m, 2H), 1.10-1.12 (m, 2H),
1.13 (t, J=7.2 Hz, 3H), 1.22-1.48 (m, 22H), 4.01 (q, J=7.2 Hz, 2H),
8.56 (s, 1H). MS: 337 (M+H)
[0221] Step 2: To a mixture of potassium-t-butoxide (3.0 g, 27.4
mmol) and toluenesulphonylmethyl isocyanide (2.7 g, 13.7 mmol) in
1,2-dimethoxyethane (20 mL) was added a solution of product of Step
1 (2.2 g, 6.5 mmol) in 1,2-dimethoxyethane (5 mL) at -60.degree. C.
and continued stirring at same temperature for 10 min. Then
reaction mixture was allowed to reach room temperature and
continued stirring for 1 h. To the reaction mixture methanol (25
mL) was added and refluxed for 15 min. The residue obtained upon
evaporation of volatiles was dissolved in 4% acetic acid (50 mL),
extracted with dichloromethane (100 mL.times.3). The combined
organic layers were washed with 10% NaHCO.sub.3 (20 mL), dried
(Na.sub.2SO.sub.4) and concentrated to give crude product as orange
liquid. The crude product was purified by silica gel column (5.0%
ethyl acetate in petroleum ether) to give crude product as yellow
liquid 1.6 g (70%) that was used in next reaction without further
purification.
[0222] Step 3: To a solution of product of Step 2 (1.3 g, 3.7 mmol)
in ethanol (12 mL) was added water (6 mL), potassium hydroxide (6.2
g, 112.2 mmol) and the reaction mass was stirred at 140.degree. C.
over a period of 16 h. The crude product obtained upon evaporation
of the solvent was diluted with water (10 mL), acidified to pH=2
(6N HCl) and extracted with ethyl acetate (100 mL.times.3). The
crude product obtained upon evaporation of the solvent was purified
by silica gel column (20% ethyl acetate in petroleum ether) to give
product as white solid 0.62 g (49.2%). mp: 75.3.degree.
C.-78.9.degree. C.
[0223] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.26-0.34
(m, 4H), 0.63-0.64 (m, 2H), 0.99-1.0 (m, 2H), 1.21-1.26 (m, 18H),
1.39 (m, 4H), 2.11 (s, 2H), 11.95 (bs, 2H exchangeable-COOH).
.sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 12.00, 15.07,
17.48, 23.28, 26.51, 27.72, 29.50, 29.72, 33.83, 36.51, 173.77 and
176.65. MS: 337 (M-H)
Example 31
Synthesis of 2,15-Dicyclopropyl-hexadecanedioic acid diamide
##STR00047##
[0225] To a solution of product of Example 1 (2.0 g, 5.9 mmol) in
dichloromethane (DCM) (20 mL) was added oxalyl chloride (1.98 mL,
23.6 mmol) and catalytic amount of DMF (0.02 mL) at ice
temperature. The ice bath was removed and reaction mixture stirred
at room temperature over a period of 1 h. The crude product
obtained upon evaporation of the solvent was diluted with DCM (10
mL), cooled to ice bath temperature and added ammoniated DCM (100
mL) drop wise fashion. Then reaction mixture was slowly allowed to
reach room temperature and stirred for 2 h. The resulting reaction
mass was filtered, washed with water (2.times.50 mL), DCM
(2.times.30 mL) and dried under suction to give product as
off-white solid 1.5 g (75%). mp: 157.0-157.9.degree. C.
[0226] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.45-0.48
(m, 4H), 0.86-0.89 (m, 4H), 1.22-1.31 (bs, 20H), 1.43-1.46 (m, 4H),
6.78 (bs, 2H, D.sub.2O exchangeable .sup.1H), 6.96 (bs, 2H,
D.sub.2O exchangeable .sup.1H). .sup.13C NMR (75 MHz, DMSO-d.sub.6)
.delta. (ppm): 13.77, 24.70, 27.79, 29.52, 29.63, 34.36 and 175.92.
MS: 337 (M+H)
Example 32
Synthesis of 1,1'-dodecane-1,12-diyldicyclopropane carbonitrile
##STR00048##
[0228] A solution of n-butyl lithium in hexane (1.8M, 1.06 L, 1.92
mol) was added drop wise to a solution of freshly distilled
diisopropylamine (280.8 mL, 1.82 mol) in dry THF (1.5 L) at
-60.degree. C. under nitrogen atmosphere. The reaction mixture was
slowly warmed to -20.degree. C. and stirred for 30 min. The
reaction mixture was re-cooled to -78.degree. C., DMPU (29.3 g,
0.22 mol) was added drop wise and stirred for 15 min. Then
cyclopropane carbonitrile (122.7 g, 1.82 mol) added drop wise over
a period 1 h and stirred for 1 h. To the reaction mixture was added
drop wise 1,12-dibromododecane (150.0 g, 0.45 mol) in dry THF (150
mL) and continued stirring at -78.degree. C. for 2 h. The reaction
mixture was slowly allowed to reach room temperature and continued
stirring over a period of 16 h. The resulting reaction mixture was
quenched with saturated NH.sub.4Cl (3.0 L) at 0.degree. C. and the
reaction mixture was extracted with ethyl acetate (1 L.times.3).
The organic phases were combined, dried over anhydrous
Na.sub.2SO.sub.4 and volatiles were evaporated under reduced
pressure to yield a pale yellow liquid which was purified through
silica gel (230-400) column (5% ethyl acetate in petroleum ether)
to give product as off-white solid. The off-white solid washed with
petroleum ether to obtain product as white solid 112 g (81.5%). mp:
45.0.degree. C.-46.9.degree. C.
[0229] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.82-0.85
(m, 4H), 1.11-1.15 (m, 4H), 1.24 (bs, 16H), 1.38-1.42 (m, 8H).
.sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 9.75, 13.62,
27.81, 28.84, 29.34, 29.41, 29.44, 34.49, and 124.00. MS: 301
(M+H)
Example 33
Preparation of (2,15-dicyclopropyl-hexadecanedioic acid monomethyl
ester
##STR00049##
[0231] To a suspension of product of Example 1 (15.0 g, 44.3 mmol)
in methanol (150 mL) was added thionyl chloride (3.2 mL, 44.3 mmol)
drop wise at ice temperature. Ice bath was removed and reaction
mixture stirred at ambient temperature over a period of 48 h.
Filtered the reaction mixture to remove unreacted starting material
(9 g) and residue obtained upon evaporation of the filtrate was
purified by column chromatography (10% ethyl acetate in petroleum
ether) to give product as a white solid 1.5 g (9.6%). mp:
59.4.degree. C.-60.5.degree. C.
[0232] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.72
(m, 4H), 1.00-1.04 (m, 4H), 1.22 (bs, 16H), 1.40-1.46 (m, 8H), 3.55
(s, 3H), 11.98 (s, 1H, --COOH, D.sub.2O exchangeable .sup.1H).
.sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 15.60, 16.45,
23.34, 23.52, 27.55, 27.66, 29.56, 29.62, 29.81, 33.58, 33, 97,
51.65, 176.01 and 182.31. MS: 351 (M-H)
Example 34
Preparation of
1-[12-(1-cyclopropylcarbamoyl-cyclopropyl)-dodecyl]-cyclopropane-carboxyl-
ic acid methyl ester
##STR00050##
[0234] To a solution of product of Example 33 (0.91 g, 2.58 mmol)
in dichloromethane (9.1 mL) was added oxalyl chloride (0.4 mL, 5.1
mmol) and catalytic amount of DMF (0.015 mL) at ice temperature.
The ice bath was removed and reaction mixture stirred at room
temperature over a period of 1 h. The crude product obtained upon
evaporation of the solvent was diluted with DCM (10 mL), cooled to
ice bath temperature added triethyl amine (1.0 mL, 7.75 mmol), and
cyclopropyl amine (1.04 g, 2.5 mmol) added in a drop wise fashion.
Then reaction mixture was slowly allowed to reach room temperature
and stirred for 16 h. The resulting reaction mass was diluted with
ethyl acetate (250 mL), washed with water (50 mL) and dried over
sodium sulphate. The residue obtained upon evaporation of the
volatiles was purified by silica gel (230-400) column (25% ethyl
acetate in petroleum ether) to give product as an off-white solid
0.79 g (79%). mp: 50.6.degree. C.-53.0.degree. C.
[0235] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.37-0.49
(m, 4H), 0.55-0.58 (m, 2H), 0.71-0.72 (m, 2H), 0.84-0.85 (m, 2H),
1.03-1.05 (m, 2H), 1.22 (bs, 20H), 1.36-1.46 (m, 4H), 2.54-2.59 (m,
1H), 3.56 (s, 3H), 7.42 (bs, 1H, --CONN--, D.sub.2O exchangeable
.sup.1H). .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 6.61,
11.78, 14.00, 15.41, 22.82, 23.47, 24.53, 25.22, 27.26, 27.38,
27.57, 29.43, 29.48, 29.52, 29.73, 33.91, 34.55, 36.08, 51.49,
175.45 and 175.79. MS: 392 (M+H)
Example 35
Preparation of
1-[12-(1-Cyclopropylcarbamoyl-cyclopropy)-dodecyl]-cyclopropane
carboxylic acid
##STR00051##
[0237] To a solution of product of Example 34 (0.7 g, 1.9 mmol) in
methanol (7 mL) was added KOH (0.36 g, 6.4 mmol) in water (2.1 mL)
and the reaction mixture was stirred at 60.degree. C. over a period
of 16 h. The crude product obtained upon evaporation of the solvent
was diluted with water (5.0 mL), acidified to pH=2 (1.5N HCl),
extracted with ethyl acetate (100 mL.times.2). The organic layer
was dried over sodium sulphate and concentrated. The residue
obtained upon evaporation of the volatiles was purified by silica
gel (230-400) column (50% ethyl acetate in petroleum ether) to give
product as white solid 0.51 g (76%). mp: 95.3.degree.
C.-97.7.degree. C.
[0238] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.37-0.45
(m, 4H), 0.52-0.57 (m, 2H), 0.63-0.66 (m, 2H), 0.83-0.86 (m, 2H),
0.98-1.01 (m, 2H), 1.22 (bs, 18H), 1.40-1.46 (m, 6H), 2.51-2.59 (m,
1H), 7.42 (s, 1H, --CONH--, D.sub.2O exchangeable 1H), 11.99 (s,
1H, --COOH, D.sub.2O exchangeable .sup.1H). .sup.13C NMR (75 MHz,
DMSO-d.sub.6) .delta. (ppm): 6.68, 14.10, 16.21, 22.92, 23.29,
24.54, 27.37, 27.49, 29.42, 29.48, 29.51, 29.70, 29.75, 33.60,
34.56, 175.76 and 181.51. MS: 376 (M-H)
Example 36
Preparation of
1-[12-(1-Methanesulfonylaminocarbonyl-cyclopropyl)-dodecyl]-cyclopropanec-
arboxylic acid methyl ester
##STR00052##
[0240] To a solution of product of Example 33 (1.6 g, 4.5 mmol) in
THF (16 mL, 10V) was added CDI (1.84 g, 8.1 mmol) and DMAP (0.06 g,
0.4 mmol) at room temperature. The reaction mixture stirred at room
temperature over a period of 16 h. To the resultant reaction
mixture was added DBU (2.27 g, 14.9 mmol), methane sulfonamide
(1.08 g, 11.3 mmol) and the resulting reaction mixture was refluxed
at 100.degree. C. over a period of 5 h. The reaction mixture was
quenched with saturated NH.sub.4Cl (20 mL), diluted with ethyl
acetate (200 mL), separated organic layer, washed with water (50
mL), brine (50 mL), dried over sodium sulphate and evaporated under
reduced pressure. The residue obtained upon evaporation of the
volatiles was purified by silica gel (230-400) column (25% ethyl
acetate in petroleum ether) to give product as white solid 1.2 g
(63%). mp: 53.5.degree. C.-54.5.degree. C.
[0241] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.63-0.73
(m, 4H), 1.02-1.08 (m, 4H), 1.23 (bs, 20H), 1.31-1.52 (m, 4H), 3.20
(s, 3H), 3.57 (s, 3H), 11.17 (s, 1H, --CONH--, D.sub.2O
exchangeable .sup.1H). .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta.
(ppm): 15.44, 15.62, 23.49, 25.78, 27.41, 27.56, 29.39, 29.52,
29.72, 33.52, 33.89, 41.45, 51.54, 173.74 and 175.88. MS: 428.0
(M-H)
Example 37
1-[12-(1-Methanesulfonylaminocarbonyl-cyclopropyl)-dodecyl]-cyclopropaneca-
rboxylic acid
##STR00053##
[0243] To a solution product of Example 36 (0.85 g, 1.97 mmol) in
methanol (8.5 mL) was added KOH (0.39 g, 7.1 mmol) in water (2.1
mL) and the reaction mixture was stirred at 60.degree. C. over a
period of 16 h. The crude product obtained upon evaporation of the
solvent was diluted with water (5.0 mL), acidified to pH=2 (1.5N
HCl), extracted with ethyl acetate (100 mL.times.2). The organic
layer was dried over sodium sulphate and concentrated. The residue
obtained upon evaporation of the volatiles was purified by silica
gel (230-400) column (25% ethyl acetate in petroleum ether) to give
product as white solid 0.55 g (67%). mp: 97.5.degree.
C.-99.4.degree. C.
[0244] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.61-0.62
(m, 4H), 0.95-0.96 (m, 4H), 1.02 (bs, 18H), 1.18-1.35 (m, 4H), 1.48
(m, 2H), 3.30 (s, 3H), 11.12 (s, 1H, --CONH--, D.sub.2O
exchangeable 1H), 11.95 (s, 1H, --COOH--, D.sub.2O exchangeable
.sup.1H). .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta. (ppm): 15.75,
16.33, 23.28, 25.77, 27.44, 29.48, 29.73, 33.51, 41.53, 173.78 and
181.77. MS: 414 (M-H)
Example 38
Preparation of
1-[12-(1-Difluoromethyl-cyclopropyl)-dodecyl]-cyclopropanecarboxylic
acid ethyl ester
##STR00054##
[0246] Step 1: A solution of dichloromethane (3 mL) and oxalyl
chloride (0.36 mL, 4.3 mmol) in a 50 mL flask was cooled in a dry
ice-acetone bath (-78.degree. C.). A solution of dimethyl sulfoxide
(0.62 mL, 8.9 mmol) dissolved in dichloromethane (2 mL) was added
to the stirred oxalyl chloride solution at -70.degree. C. The
reaction mixture was stirred for 5 minutes, and a solution of
product of Step 1, Example 13 (0.7 g, 1.9 mmol) in dichloromethane
(2 mL) was added drop wise over a period of 5 minutes. Stirring was
continued for an additional 45 min at -70.degree. C. and
triethylamine (2.7 mL, 19.8 mol) was added. The reaction mixture
was stirred for 5 min and then allowed to warn to room temperature.
After 1 hour, water (100 mL) was added and the aqueous layer was
extracted with additional dichloromethane (200 mL). The organic
layer was combined, washed with brine (200 mL) and dried over
anhydrous sodium sulphate. The crude product obtained upon
evaporation of the solvent was purified by silica gel column (2.0%
ethyl acetate in petroleum ether) to give product as a yellow
liquid 0.6 g (87%). The aldehyde seems unstable on storage and was
used in next step quickly.
[0247] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.68-0.71
(m, 2H), 0.94-0.99 (m, 2H), 1.01-1.04 (m, 2H), 1.11-1.13 (m, 2H),
1.14-1.17 (m, 3H), 1.23 (bs, 18H), 1.31-1.46 (m, 6H), 4.02 (q,
J=7.2 Hz, 3H), 8.56 (s, 1H).
[0248] Step 2: To a solution of product of Step 1 (0.6 g, 1.71
mmol) in dry THF (6.0 mL) was added diethylamino-sulfur trifluoride
(1.31 mL, 8.5 mmol) drop wise at room temperature and the reaction
mixture was stirred at same temperature over a period of 16 h. The
resulting reaction mass was cooled to ice both temperature and
quenched with water (20 mL) and extracted with ethyl acetate (200
mL). the organic layer was washed with 5% sodium bicarbonate (100
mL), brine (100 mL) and dried over sodium sulphate. The crude
product obtained upon evaporation of the solvent was purified by
silica gel column (2.0% ethyl acetate in petroleum ether) to give
product as a yellow oil 0.5 g (79%).
[0249] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.48-0.49
(m, 2H), 0.62-0.64 (m, 2H), 0.68-0.71 (m, 2H), 1.01-1.04 (m, 2H),
1.50 (t, J=7.2 Hz, 3H), 1.23 (bs, 16H), 1.35-1.48 (bs, 8H), 4.02
(q, J=7.2 Hz, 3H), 5.66 (t, J=56.7 Hz, 1H).
Example 39
Synthesis of
1-[12-(1-Difluoromethyl-cyclopropyl)-dodecyl]-cyclopropanecarboxylic
acid
##STR00055##
[0251] To a solution of potassium hydroxide (0.63 g, 11.2 mol) in
90% ethanol (5 mL) was added product of Example 38 (0.5 g, 1.3
mmol) at 40.degree. C. and the reaction mass was stirred at
40.degree. C. over a period of 16 h. The crude product obtained
upon evaporation of the solvent was diluted with water (5 mL),
acidified to pH=2 (1N HCl) and extracted with ethyl acetate (100
mL.times.3). The crude product obtained upon evaporation of the
solvent was purified by silica gel column (5% ethyl acetate in
petroleum ether) to give product as white solid 0.32 g (69.5%). mp:
54.0.degree. C.-54.8.degree. C.
[0252] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.48-0.49
(m, 2H), 0.64-0.65 (m, 4H), 1.00-1.02 (m, 2H), 1.23 (bs, 16H), 1.40
(bs, 8H), 5.67 (t, J=56.7 Hz, 1H), 11.97 (bs, 2H
exchangeable-COOH). .sup.13C NMR (75 MHz, DMSO-d.sub.6) .delta.
(ppm): 7.88, 7.94, 8.00, 15.01, 21.89, 22.20, 22.52, 23.29, 26.62,
27.69, 29.36, 29.45, 29.70, 29, 82, 30.99, 33.83, 116.87, 116.90,
120.03, 120.06, 123.18, 123.21, and 176.58. MS: 343 (M-H)
Example 40
Preparation of
1,1-dodecane-1,12-diylbis[N-(2-hydroxyethyl)cyclopropanecarboxamide]
##STR00056##
[0254] To the product of Example 10 (2.0 g, 5.0 mmol) in a round
bottomed flask, ethanolamine (10 mL) was added at room temperature.
The reaction mixture was heated at 150.degree. C. over a period of
18 h. The resulting reaction mixture was diluted with ethyl acetate
(100 mL), washed with water (2.times.50 mL) and dried over sodium
sulphate. The crude product obtained upon evaporation of the
volatiles was purified through silica gel (230-400) column (50%
ethyl acetate in petroleum ether) to obtain pure compound A as
white solid (0.5 g, 23%) and pure compound B as pale yellow solid
(0.75 g, 36%). Compound A: mp: 112.5.degree. C.-113.5.degree. C. 1H
NMR (300 MHz, DMSO-d6) .delta. (ppm): 0.46-0.48 (m, 4H), 0.86-0.89
(m, 4H), 1.22-1.28 (bs, 20H), 1.45-1.48 (m, 4H), 3.07-3.13 (m, 4H),
3.32-3.38 (m, 4H), 4.60 (t, J=5.7 Hz, 2H exchangeable-OH), 7.37 (t,
J=5.7 Hz, 2H). 13C NMR (75 MHz, DMSO-d6) .delta. (ppm): 13.51,
24.98, 27.62, 29.48, 29.60, 34.23, 42.35, 60.46 and 173.63. MS: 425
(M+H)
Example 41
Preparation of
1-{12-[1-(2-Hydroxy-ethylcarbamoyl)-cyclopropyl]-dodecyl}-cyclopropane
carboxylic acid ethyl ester
##STR00057##
[0256] The target compound (B) was prepared as described in Example
40.
[0257] mp: 42.2.degree. C.-44.3.degree. C. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. (ppm): 0.60-0.63 (m, 2H), 0.65-0.68 (m, 2H),
1.19-1.94 (m, 4H), 1.21-1.24 (m, 3H), 1.27 (bs, 16H), 1.43-1.57 (m,
8H), 2.06 (bs, 1H, exchangeable-OH), 3.43-3.48 (m, 2H), 3.72-3.76
(m, 2H), 4.11 (q, J=7.2 Hz, 2H), 6.21 (bs, 2H). .sup.13C NMR (75
MHz, CDCl3) .delta. (ppm): 14.15, 14.30, 15.36, 23.53, 24.61,
27.43, 27.57, 29.48, 29.54, 29.73, 29.78, 33.90, 34.47, 42.73,
60.21, 62.44, 175.44 and 173.63. MS: 410 (M+H)
Example 42
Synthesis of
1-{12-[1-(2-hydroxy-ethylcarbamoyl)-cyclopropyl]-dodecyl}-cyclopropane
carboxylic acid
##STR00058##
[0259] To a solution of potassium hydroxide (0.63 g, 11.2 mol) in
90% ethanol (5 mL) was added product B of Example 40 (0.5 g, 1.3
mmol) at 40.degree. C. and the reaction mass was stirred at
40.degree. C. over a period of 2 h. The crude product obtained upon
evaporation of the solvent was diluted with water (10 mL),
acidified to pH=2 (1.5 N HCl). The white precipitate obtained upon
acidification was filtered, washed with petroleum ether to obtain
the product as white solid (300 mg, 65.2%). mp: 81.1.degree.
C.-83.8.degree. C.
[0260] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. (ppm): 0.46 (bs,
2H), 0.64-0.65 (m, 2H), 0.86-0.87 (m, 2H), 0.99-1.00 (m, 2H), 1.22
(bs, 18H), 1.40-1.48 (m, 8H), 3.07-3.13 (m, 2H), 3.33-3.36 (m, 2H),
4.61 (t, J=5.1 Hz, 1H exchangeable-OH), 7.39 (bs, 1H,
exchangeable-CONH), 11.99 (s, 1H, exchangeable-COOH). .sup.13C NMR
(75 MHz, DMSO-d.sub.6) .delta. (ppm): 13.52, 15.02, 23.30, 24.97,
27.62, 27.69, 29.47, 29.60, 29.69, 33.82, 34.22, 42.34, 60.45,
173.62 and 176.58. MS: 382 (M+H)
Example 43
AMPK Activation Assay
[0261] The assay protocol used for screening compounds is described
below:
[0262] Reagents: [0263] PathScan Phospho-AMPK (Thr172) Sandwich
ELISA Antibody Pair (Cell Signaling Cat. No. 7955) [0264] DuoSet IC
Human/Mouse/Rat Total AMPK1 (R&D Systems, Cat.#DYC3197-5)
[0265] Lysis Buffer (Cell Signaling Technology, Cat. No. 9803)
diluted to 1.times. in distilled water containing 1 mM PMSF. [0266]
3,3',5,5'-Tetramethylbenzidine (TMB) Substrate (Sigma Cat No.
T8665)
[0267] Day 1: HepG2 cells were seeded at a density of 50,000 cells
per well in a 24-well plate in DMEM Low glucose (5 mM) (Sigma)
containing 10% FCS (Invitrogen) with 1 nM insulin and incubated for
6 hours at 37.degree. C. After 6 hrs of incubation, the cells were
rinsed with starvation media (DMEM Low Glucose without FCS) and 0.5
mL of starvation media was added and incubated overnight at
37.degree. C.
[0268] Day 2: Stock concentrations of the compounds (along with
standard) were prepared in 100% DMSO and were diluted to the
desired concentrations in serum-free DMEM Low glucose media. The
medium was aspirated from the wells and 0.5 mL of the medium
containing the compounds were added to the appropriate wells and
the plate was incubated for 6 hours at 37.degree. C. The media was
aspirated and the cells were rinsed twice with cold PBS. 250 .mu.L
of 1.times. lysis buffer was added to all the wells and incubated
for 15 minutes on ice. The samples were centrifuged at 2000 g for 5
minutes and the supernatant was collected. Protein concentration in
the lysate was quantified by using the BCA assay (Pierce
Biotech).
Protocol for phopsho-AMPK ELISA
Coating Procedure:
[0269] Day 1: The capture antibody was diluted and coated on a
96-well plate (100 .mu.L per well). The plate was sealed and
incubated overnight at 4.degree. C.
[0270] Day 2: Each well was aspirated and washed 4.times. with wash
buffer. The plates were then blocked by adding 150 .mu.L of Block
buffer to each well. The plate was then incubated at 37.degree. C.
for 2 hours. Each plate was washed 4 times with wash buffer and 100
.mu.L of the lysate was transferred to an ELISA plate, covered, and
incubated at 37.degree. C. for 2 hours. The contents were discarded
and the plate was washed 4 times with wash buffer. After each wash,
the residual solution was removed by striking the plate on fresh
towels. 100 .mu.L of the detection antibody was then added to each
well and the plate was incubated for 1 hour at 37.degree. C. The
plate was then washed with wash buffer as earlier and 100 .mu.L of
horse radish peroxidase-linked secondary antibody was added to each
well and the plate was then incubated for 30 minutes at 37.degree.
C. The wash procedure was repeated and 100 .mu.L of substrate
solution was added to all the wells and incubated for a further 10
minutes at 37.degree. C. 100 .mu.L of Stop solution was added and
the absorbance was read at 450 nm in a micro plate reader.
Protocol for Total AMPK ELISA
Coating Procedure:
[0271] Day 1: The capture antibody was diluted to a working
concentration of 4 .mu.g/mL in PBS. A 96-well microplate (Maxisorp)
was coated with 100 .mu.L per well of the diluted Capture antibody.
The plate was sealed and incubated overnight at 4.degree. C.
[0272] Day 2: Each well was aspirated and washed 3 times with wash
buffer. The plates were then blocked by adding 300 .mu.L of Block
buffer to each well. The plate was then incubated at room
temperature for 2 hours. Each plate was washed 3 times with wash
buffer and 100 .mu.L of the lysate was transferred to the ELISA
plate, covered, and incubated at room temperature for 2 hours. The
contents were discarded and the plate was washed 3 times with wash
buffer. After each wash, the residual solution was removed by
striking the plate on fresh towels. 100 .mu.L of the detection
antibody was then added to each well and the plate was incubated
for 2 hours at room temperature. The plate was then washed with
wash buffer as earlier and 100 .mu.L of horse radish
peroxidase-linked secondary antibody was added to each well and the
plate was incubated for 20 minutes at room temperature. The wash
procedure was repeated and 100 .mu.L of substrate solution was
added to all the wells and incubated for a further 20 minutes at
room temperature. 50 .mu.L of Stop solution was added and the
absorbance was read at 450 nm in a microplate reader.
[0273] Data Analysis--Data analysis was performed and was expressed
in Relative absorbance per mg protein and the ratio of
phosphorylated AMPK to total AMPK levels was calculated.
[0274] Compounds that showed an increase of .gtoreq.40% at <60
.mu.m concentration in AMPK activation assay and/or showed a
reduction in triglycerides as a consequence of inhibition of fatty
acid synthesis are considered active.
[0275] In vivo Efficacy: The in vivo efficacy of compounds of
general Formula I can be evaluated in diabetes and dyslipidemia
using animals models known in literature. Db/db mouse model is the
most commonly used diabetic dyslipidemia model. These animals have
high plasma glucose, insulin and triglycerides and exhibit severe
insulin resistance (ref--Young A A, Gedulin B R, Bhaysar S, Bodkin
N, Jodka C, Hansen B, Denaro M. Diabetes. 1999 May; 48(5):1026-34;
Yasuda N, Inoue T, Nagakura T, Yamazaki K, Kira K, Saeki T, Tanaka
I. J Pharmacol Exp Ther. 2004 August; 310(2): 614-9.
Example 44
In Vivo Efficacy of Representative Compounds in Diabetic Mice
[0276] The model described below can be used to evaluate potential
of representative compounds of generic Formula I to treat diabetes
and dyslipidemia.
[0277] Male obese diabetic C57BLKS db/db mice (6 weeks old) were
procured from the Jackson Laboratory (Bar Harbor, Me.) and were
maintained on LabDiet 5K52 ad libitum with access to water. Animals
were kept in a temperature-controlled room on a 12:12-h light-dark
cycle. Experiments were performed on 6-9 week old mice.
[0278] Blood samples were collected after a 6 h fast on day 0
(baseline) and at the end week 1 and week 2 (day 7 and day 14) for
analysis of glucose, triglycerides, total cholesterol, insulin and
free fatty acid levels. Body weight and feed intake was measured
twice weekly. Before initiation of the treatment, the animals were
randomized based on plasma glucose levels into treatment groups
(n=8 per group). The animals were administered the test compounds
in 0.5% carboxymethylcellulose in water at the mentioned doses for
two weeks by oral gavage. The control animals received the vehicle
(0.5% carboxymethylcellulose in water) alone. Compound
administration was continued till day 16.
Measurement of Plasma Parameters
[0279] Plasma glucose (PG) and triglycerides (TG) were measured
spectrophotometrically in a microplate reader using reagent kits
(Labkit, Merck) according to the manufacturer's instructions.
Plasma insulin was measured by ELISA using a Rat/Mouse kit from
Millipore (Cat. No. EZRMI-13K).
Data Analysis
[0280] The percentage reduction was calculated according to the
following formula
1 - Final day treated value / Day 0 treated value Final day control
value / Day 0 Control value .times. 100 ##EQU00001##
Statistical analysis was carried out by one way ANOVA followed by
Dunnett's test using GraphPad Prism.
[0281] Determination of Homeostatic Model Assessment of Insulin
Resistance (HOMA)--
[0282] The HOMA index for insulin resistance was calculated using
the formula--
I0.times.G0/405
Where I0 is the fasting insulin (.mu.U/ml), G0 is the fasting
glucose in mg/dL. A factor of 6.945 was used for the conversion of
insulin pmol/L to .mu.lU/L).
[0283] The table below summarizes the effect on plasma glucose,
triglycerides, and insulin resistance after 1-2 weeks of treatment
with representative compounds of Formula I. Example numbers denote
the products of examples given above. The data presented in the
table is representative and should not be read as limiting the
invention to the specific examples listed in the Table.
TABLE-US-00001 Plasma Insulin resistance Plasma glucose
Triglycerides HOMA index (% change (% change (% change Dose
compared to compared to compared to Example (mg/kg) vehicle)
vehicle) vehicle) 4 50 -51 -54 1 10 -52 -65 12 20 -55 -28 -89 15 15
-56 -35 -92 10 15 -51 -23 35 15 -30 37 15 -59
[0284] Although exemplary embodiments of the invention have been
described using specific terms, devices, and methods, such
description is for illustrative purposes only. The words used are
words of description rather than of limitation. It is to be
understood that changes and variations may be made by those of
ordinary skill in the art without departing from the spirit or
scope of the present invention, which is set forth in the following
claims. In addition, it should be understood that aspects of the
various embodiments may be exchanged in whole or in part.
* * * * *