Note: Descriptions are shown in the official language in which they were submitted.
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DESCRIPTION
MATERIALS AND METHODS FOR THE TREATMENT OF DIABETES,
HYPERLIPIDEMIA, HYPERCHOLESTEROLEMIA, AND ATHEROSCLEROSIS
Background of the Invention
Diabetes is one of the most prevalent chronic disorders worldwide with
significant personal and financial costs for patients and their families, as
well as for
society. Different types of diabetes exist with distinct etiologies and
pathogeneses.
For example, diabetes mellitus is a disorder of carbohydrate metabolism,
characterized by hyperglycemia and glycosuria and resulting from inadequate
production or utilization of insulin.
Noninsulin-dependent diabetes mellitus (I~TIDDM), often referred to as Type II
diabetes, is a form of diabetes that occurs predominantly in adults who
produce
adequate levels of insulin but who have a defect in insulin-mediated
utilization and
metabolism of glucose in peripheral tissues. Overt NIDDM is characterized by
three
major metabolic abnormalities: resistance to insulin-mediated glucose
disposal,
impairment of nutrient-stimulated insulin secretion, and overproduction of
glucose by
the liver. It has been shown that for some people with diabetes a genetic
predisposition results from a mutation in the genes) coding for insulin and/or
the
insulin receptor and/or insulin-mediated signal transduction factor(s),
thereby
resulting in ineffective insulin and/or insulin-mediated effects thus
impairing the
utilization or metabolism of glucose.
For people with Type II diabetes, insulin secretion is often enhanced,
presumably to compensate for insulin resistance. Eventually, however, the B-
cells
fail to maintain sufficient insulin secretion to compensate for the insulin
resistance.
Mechanisms responsible for the B-cell failure have not been identified, but
may be
related to the chronic demands placed on the B-cells by peripheral insulin
resistance
and/or to the effects of hyperglycemia. The B-cell failure could also occur as
an
independent, inherent defect in "pre-diabetic" individuals.
NIDDM often develops from certain at risk populations. One such population
is individuals with polycystic ovary syndrome (PCOS). PCOS is the most common
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endocrine disorder in women of reproductive age. This syndrome is
characterized by
hyperandrogenism and disordered gonadotropin secretion producing oligo- or
anovulation. Recent prevalence estimates suggest that 5-10% of women between
18-
44 years of age (about 5 million women, according to the 1990 census) have the
full-
y blown syndrome of hyperandrogenism, chronic anovulation, and polycystic
ovaries.
Despite more than 50 years since its original description, the etiology of the
syndrome
remains unclear. The biochemical profile, ovarian morphology, and clinical
features
are non-specific; hence, the diagnosis remains one of exclusion of disorders,
such as
androgen-secreting tumors, Cushing's Syndrome, and late-onset congenital
adrenal
hyperplasia. PCOS is associated with profound insulin resistance resulting in
substantial hyperinsulinemia. As a result of their insulin resistance, PCOS
women are
at increased risk to develop NIDDM.
TtIDDM also develops from the at risk population of individuals with
gestational diabetes mellitus (GDM). Pregnancy normally is associated with
progressive resistance to insulin-mediated glucose disposal. In fact, insulin
sensitivity
is lower during late pregnancy than in nearly all other physiological
conditions. The
insulin resistance is thought to be mediated in large part by the effects of
circulating
hormones such as placental lactogen, progesterone, and cortisol, all of which
are
elevated during pregnancy. In the face of the insulin resistance, pancreatic B-
cell
responsiveness to glucose normally increases nearly 3-fold by late pregnancy,
a
response that serves to minimize the effect of insulin resistance on
circulating glucose
levels. Thus, pregnancy provides a major "stress-test" of the capacity for B-
cells to
compensate for insulin resistance.
Other populations thought to be at risk for developing TIIDDM include
persons with Syndrome X; persons with concomitant hyperinsulinemia; persons
with
insulin resistance characterized by hyperinsulinemia and by failure to respond
to
exogenous insulin; and persons with abnormal insulin andlor evidence of
glucose
disorders associated with excess circulating glucocorticoids, growth hormone,
catecholamines, glucagon, parathyroid hormone, and other insulin-resistant
conditions.
Failure to treat l~IIDDM can result in mortality due to cardiovascular disease
and in other diabetic complications including retinopathy, nephropathy, and
peripheral neuropathy. There is a substantial need for a method of treating at
risk
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populations such as those with PCOS and GDM in order to prevent or delay the
onset
of NIDDM thereby bringing relief of symptoms, improving the quality of life,
preventing acute and long-term complications, reducing mortality and treating
accompanying disorders of the populations at risk for NIDDM.
For many years, treatment of NIDDM has involved a program aimed at
lowering blood sugar with a combination of diet and exercise. Alternatively,
treatment
of NIDDM can involve oral hypoglycemic agents, such as sulfonylureas alone or
in
combination with insulin injections. Recently, alpha-glucosidase inhibitors,
such as a
carboys, have been shown to be effective in reducing the postprandial rise in
blood
glucose (Lefevre, et al., Drugs 1992; 44:29-38). In Europe and Canada another
treatment used primarily in obese diabetics is metformin, a biguanide.
Compounds useful in the treatment of the various disorders discussed above,
and methods of making the compounds, are known and some of these are disclosed
in
U.S. Pat. Nos. 5,223,522 issued Jun. 29, 1993; 5,132,317 issued Jul. 12, 1992;
5,120,754 issued Jun. 9, 1992; 5,061,717 issued Oct. 29, 1991; 4,897,405
issued Jan.
30, 1990; 4,873,255 issued Oct. 10, 1989; 4,687,777 issued Aug. 18, 1987;
4,572,912
issued Feb. 25, 1986; 4,287,200 issued Sep. l, 1981; 5,002,953, issued Mar.
26, 1991;
U.S. Pat. Nos. 4,340,605; 4,438,141; 4,444,779; 4,461,902; 4,703,052;
4,725,610;
4,897,393; 4,918,091; 4,948,900; 5,194,443; 5,232,925; and 5,260,445; WO
91107107; WO 92102520; WO 94/01433; WO 89/08651; and JP Kokai 69383/92. The
compounds disclosed in these issued patents and applications are useful as
therapeutic
agents for the treatment of diabetes, hyperglycemia, hypercholesterolemia, and
hyperlipidemia. The teachings of these issued patents are incorporated herein
by
reference in their entireties.
Drug toxicity is an important consideration in the treatment of humans and
animals. Toxic side effects resulting from the administration of drugs include
a
variety of conditions that range from low-grade fever to death. Drug therapy
is
justified only when the benefits of the treatment protocol outweigh the
potential risks
associated with the treatment. The factors balanced by the practitioner
include the
qualitative and quantitative impact of the drug to be used as well as the
resulting
outcome if the drug is not provided to the individual. Other factors
considered
include the physical condition of the patient, the disease stage and its
history of
progression, and any known adverse effects associated with a drug.
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Drug elimination is typically the result of metabolic activity upon the drug
and
the subsequent excretion of the drug from the body. Metabolic activity can
take place
within the vascular supply and/or within cellular compartments or organs. The
liver is
a principal site of drug metabolism. The metabolic process can be categorized
into
synthetic and nonsynthetic reactions. In nonsynthetic reactions, the drug is
chemically altered by oxidation, reduction, hydrolysis, or any combination of
the
aforementioned processes. These processes are collectively referred to as
Phase I
reactions.
In Phase II reactions, also known as synthetic reactions or conjugations, the
parent drug, or intermediate metabolites thereof, are combined with endogenous
substrates to yield an addition or conjugation product. Metabolites formed in
synthetic reactions are, typically, more polar and biologically inactive. As a
result,
these metabolites are more easily excreted via the kidneys (in urine) or the
liver (in
bile). Synthetic reactions include glucuronidation, amino acid conjugation,
acetylation, sulfoconjugation, and methylation.
One of the drugs used to treat Type II diabetes is troglitazone. The major
side
effects of troglitazone are nausea, peripheral edema, and abnormal liver
function.
Other reported adverse events include dyspnea, headache, thirst,
gastrointestinal
distress, insomnia, dizziness, incoordination, confusion, fatigue, pruritus,
rash,
alterations in blood cell counts, changes in serum lipids, acute renal
insufficiency, and
dryness of the mouth. Additional symptoms that have been reported, for which
the
relationship to troglitazone is unknown, include palpitations, sensations of
hot and
cold, swelling of body parts, skin eruption, stroke, and hyperglycemia.
Accordingly,
forms of glitazones which have fewer, or no, adverse effects (i.e., less
toxicity) are
desirable.
The principal difference between the compounds of the present invention and
related compounds is the presence of a carboxyl group, either OOC- or COO-,
directly attached to the 4-position of the phenyl ring. In the literature,
thiazolidinediones having similar therapeutic properties have an ether
function at the
4-position of the phenyl ring instead of a carboxyl group.
The presence of the carboxyl group has significant consequences for the
biological behavior of these new compounds. The present compounds are
primarily
metabolized by hydrolytic enzymatic systems, whereas compounds having an ether
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function are metabolized only by oxidative enzymes. Hydrolytic enzymatic
systems
are ubiquitous, non-oxidative, not easily saturable, and non-inducible, and,
therefore,
reliable. By contrast, oxidative systems are mediated by the P-450 isozymes.
These
systems are localized, mainly, in the liver, saturable and inducible (even at
low
5 concentrations of therapeutic compounds) and therefore are highly
unreliable.
The compounds of the subject invention do not rely on saturable hepatic
systems fox their metabolism and elimination, whereas the prior art compounds
exert
a heavy bio-burden on hepatic functions, especially in the presence of other
drugs that
rely on similar enzymes for detoxification. Thus, the present compounds have a
much
more desirable toxicity profile than prior art compounds, especially when
considering
liver toxicity and potentially fatal drug-drug interactions.
Upon metabolism by plasma and tissue esterases, the compounds of this
invention are hydrolyzed into 2 types of molecules: 1) an alcohol or a phenol,
and 2) a
carboxylic acid. Therefore, any compound that yields compound 1, compound 2,
compound 3, or compound 4, as defined in Table I, as a primary metabolite
falls
under the definition of this invention. This concept is illustrated in Figure
1, taking
compound 9 (of Table I) and compound I45 (of Table X) as specific examples of
compounds giving I and 3, respectively, upon non-oxidative metabolism by
esterases.
Brief Summary of the Invention
The subject invention provides materials and methods for the safe and
effective treatment of diabetes, hyperlipidemia, hypercholesterolemia, and
atherosclerosis. In a preferred embodiment, the subject invention provides
therapeutic
compounds for the treatment of diabetes. The compounds of the subject
invention can
be used to treat at-risk populations, such as those with PCOS and GDM, in
order to
prevent or delay the onset of NIDDM thereby bringing relief of symptoms,
improving
the quality of life, preventing acute and long-term complications, reducing
mortality
and treating accompanying disorders.
Advantageously, the subject invention provides compounds that are readily
metabolized by the physiological metabolic drug detoxification systems.
Specifically,
in a preferred embodiment, the therapeutic compounds of the subj ect invention
contain an ester group, which does not detract from the ability of these
compounds to
provide a therapeutic benefit, but which makes these compounds more
susceptible to
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degradation by hydrolases, particularly serum and/or cytosolic esterases. The
subject
invention further provides methods of treatment comprising the administration
of
these compounds to individuals in need of treatment for Type II diabetes,
hyperlipidemia, hypercholesterolemia, and atherosclerosis.
In a further embodiment, the subject invention pertains to the breakdown
products that are formed when the therapeutic compounds of the subject
invention are
acted upon by esterases. These breakdown products can be used as described
herein
to monitor the clearance of the therapeutic compounds from a patient.
In yet a further embodiment, the subject invention provides methods for
synthesizing the therapeutic compounds of the subj ect invention.
Brief Description of the Figures
Figure 1 depicts exemplary metabolic breakdown products resulting from the
actions of esterases on compounds of the invention.
Figures 2-3 provide an exemplary synthetic scheme for compounds 1 through
4 (of Table I). These compounds can be conveniently prepared by the
I~noevenagel
reaction between an aldehyde and thiazolidine-2,4-dione using, for example,
sodium
acetate in acetic anhydride, or piperidine and benzoic acid in methylene
chloride as a
reaction medium.
Figure 4 illustrates an alternative reaction scheme for the production of
compound 1 (of Table I). In this reaction scheme, para-anisidine undergoes a
diazotation reaction with sodium nitrite and hydrochloric acid. The diazonium
chloride salt undergoing, in turn, a radicalar reaction with methyl acrylate
and then a
cyclization reaction with thiourea, the product of which is hydrolyzed to the
thiazolidinedione molecule.
Figure 5 shows an exemplary synthetic scheme for the compounds described
in Table I (compounds 5 to 32). These compounds can be made via an
esterification
reaction between 1 or 2 and an appropriately substituted carboxylic acid, or
between 3
or 4 and an appropriately substituted alcohol.
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Figure 6 depicts the synthesis of the 4-oxazoleacetic acid and the 4-
oxazoleethanol moiety starting from aspartic acid derivatives in which RZ and
R3 are
methyl or hydrogen.
Figure 7 describes the synthesis of the 4-oxazolecarboxylic acid and 4
oxazolemethanol groups. The synthesis starts from ethyl acetoacetate in which
a 2
amino-group is introduced via oxime formation followed by reduction with zinc
powder. The synthesis then proceeds as before, where the Rl group is
introduced by
acylating the amino group, followed by cyclization with sulfuric acid in ethyl
acetate,
and finally ester cleavage or reduction to the alcohol.
Figure 8 shows how steric hindrance can be introduced under the form of
methyl groups on the 4-methanol moiety. Starting from pentane-2,4-dione and
following the same synthetic sequence as in Figure 7 leads to the 4-
acetyloxazole
compounds which can be reduced by sodium borohydride to the 4-(1-
ethyl)oxazole,
or which can be transformed to 4-(2-hydroxy-2-propyl) oxazole with a methyl
Grignard reagent such as methyl magnesium iodide.
Figure 9 illustrates an alternative synthetic scheme wherein condensation of a
thioamide with methyl 4-bromo-3-oxopentanoate gives methyl 4-thiazoleacetate.
Ester cleavage with lithium hydroxide or reduction with lithium aluminum
hydride
gives the corresponding acid or the alcohol, respectively.
Figures 10-17 depict the synthesis of compounds 105 to 224 in Tables VI to
XVII. These compounds contain an amino acid or an amino alcohol as part of
their
structure.
Figure 18 provides an exemplary synthetic pathway for compounds 225 to
242 (Table XVIII). These compounds are oxazoline-4-carboxylic acid types of
compounds. Their synthesis (Figure 18) starts from serine (RS=H) or from
threonine
(RS=CH3) benzyl ester. The ester is coupled with an alkyl or an arylcarboxylic
acid
using for example EDC as a coupling agent. The serine or threonine group then
cyclizes into an oxazoline upon treatment with thionyl chloride. Coupling with
5-(4-
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hydroxybenzyl)thiazolidine-2,4-dione using DCC/DMAP/methylene chloride gives
compounds 225 to 242.
Figures 19-20 illustrate the activity of representative compounds on senun
glucose and insulin levels in non-insulin dependent diabetic mellitus (NIDDM)
KK-
Ay male mice. Post-treatment data for each group were transferred to a
percentage of
pretreatment values and unpaired Student's t test was used for comparison
between
vehicle and test substance treated groups. Results show a significant
reduction of
both serum glucose and serum insulin relative to the vehicle control group.
Reduction
in serum glucose and serum insulin levels were comparable to the reduction
observed
in the troglitazone-treated animals. The results are also presented in Table
XXI.
Figure 21 shows exemplary compounds of Formula IB.
Figures 22-28 show exemplary synthesis schemes to produce compounds of
Formula 1B.
Figures 29-38 provide synthetic pathways for compounds of Formulas II-IX.
Figures 39-40 depict compounds excluded from the scope of the subject
invention.
Brief Description of the Tables
Tables I-XXII depict various compounds of the invention. The term "db"
indicates a double bond between P and Q.
Table XXIII illustrates the effects of exemplary compounds on serum glucose
and insulin levels in 1VIDDM mice.
Detailed Disclosure of the Invention
The subject invention provides materials and methods for the treatment of
non-insulin dependent diabetes mellitus (IVIDDM), hyperlipidemia,
hypercholesterolemia, and atherosclerosis. Advantageously, the therapeutic
compounds of the subject invention (identified in Formulae II-IX) are stable
in
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storage but have a shorter half life in the physiological environment than
other drugs
which are available for treatment of diabetes; therefore, the compounds of the
subject
invention can be used with a lower incidence of side effects and toxicity,
especially in
patients having elevated liver function or compromised liver function.
In a preferred embodiment of the subject invention, therapeutic compounds
are provided which are useful in the treatment of diabetes, hyperlipidemia,
hypercholesterolemia, and atherosclerosis and which contain an ester group
which is
acted upon by esterases thereby breaking down the compound and facilitating
its
efficient removal from the treated individual. In a preferred embodiment the
therapeutic compounds are metabolized by the Phase I drug detoxification
system and
are exemplified by the compounds of Formulae II-IX.
In various embodiments of the subject invention, therapeutic compounds of
the subject invention may, optionally, exclude compounds of Formulae IA and
IB.
However, the compounds of the instant invention (identified in Formulae II
through
IX) do represent a new class of chemical compounds having therapeutic
properties for
the treatment of type-II diabetes mellitus, atherosclerosis,
hypercholesterolemia, and
hyperlipidemia.
FORMULA IA
P Q O
D2
i\D Ds B A
6
X E
O
For compounds of Formula IA:
A and B may be the same or different and are CHZ, CO, N, NO, NH, SOo_2, or
O;
D1-D6 can be the same or different and are CH, N, S, or O;
E can be a substituent attached to one or more of the atoms located at Dl-D6;
P and Q can be a double bond; or
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P, Q, and E can be the same or different and are a moiety selected from the
group consisting of H, CI_lo alkyl, substituted alkyl groups, substituted or
unsubstituted carboxylic acids, substituted or unsubstituted carboxylic
esters, halogen,
carboxyl, hydroxyl, phosphate, phosphonate, aryl, CN, OH, COOH, NOz, NHz,
SOz_4,
5 Ci_zo heteroalkyl, Cz_zo alkenyl, alkynyl, akynyl-aryl, alkynyl-heteroaryl,
aryl, C1_zo
alkyl-aryl, Cz_zo alkenyl-aryl, heteroaryl, C1_zo alkyl-heteroaryl, Cz_zo
alkenyl-
heteroaryl, cycloalkyl, heterocycloalkyl, Cl_zo alkyl-heteroycloalkyl, and
CI_zo alkyl-
cycloalkyl, any of which may be, optionally, substituted with a moiety
selected from
the group consisting of C1_6 alkyl, halogen, OH, NHz, CN, NOz, COOH, or SOz~.
10 Exemplary heterocyclic groups include, but are not limited to, morpholine,
triazole,
imidazole, pyrrolidine, piperidine, piperazine, pyrrole, dihydropyridine,
aziridine,
thiazolidine, thiazoline, thiadiazolidine or thiadiazoline.
Substituted carboxylic acids, substituted carboxylic esters, and substituted
alkyl groups can be substituted at any available position with a moiety
selected from
the group consisting of C1_lo alkyl, halogen, CN, OH, COOH, NOz, NHz, SOz.~,
C1_zo
heteroalkyl, Cz_zo alkenyl, alkynyl, akynyl-aryl, alkynyl-heteroaryl, aryl, Ci-
zo alkyl-
aryl, Cz_zo alkenyl-aryl, heteroaryl, C1_zo alkyl-heteroaryl, Cz_zo alkenyl-
heteroaryl,
cycloalkyl, heterocycloalkyl, C1_zo alkyl-heteroycloalkyl, and Cl_zo alkyl-
cycloalkyl,
any of which may be, optionally, substituted with a moiety selected from the
group
consisting of C1_6 alkyl, halogen, OH, NHz, CN, NOz, COOH, or SOz.~. Exemplary
heterocyclic groups include, but are not limited to, morpholine, triazole,
imidazole,
pyrrolidine, piperidine, piperazine, pyrrole, dihydropyridine, aziridine,
thiazolidine,
thiazoline, thiadiazolidine, and thiadiazoline.
X is -OH, -COOH, or a substituted carboxylic group having the carboxyl
moiety OOC- or COO- directly attached to the phenyl ring of the compound of
Formula 1. The carboxylic acid group can be substituted with a moiety selected
from
the group consisting of alkyloxycarbonyl, alkylcarbonyloxy, aryloxycarbonyl,
arylcarbonyloxy, heteroalkyloxycarbonyl, heteroalkylcarbonyloxy, heteroaryl
oxycarbonyl, and heteroarylcarbonyloxy each of which is, optionally,
substituted with
Ci_lo alkyl, CN, COOH, NOz, NHz, SOz~, C1_zo heteroalkyl, Cz_zo alkenyl,
alkynyl,
akynyl-aryl, alkynyl-heteroaryl, aryl, C1_zo alkyl-aryl, Cz_zo alkenyl-aryl,
heteroaryl,
C1_zo alkyl-heteroaryl, Cz-zo alkenyl-heteroaryl, cycloalkyl,
heterocycloalkyl, Cl_zo
alkyl-heteroycloalkyl, and C1_zo alkyl-cycloalkyl, any of which may be,
optionally,
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substituted with a moiety selected from the group consisting of Cl_G alkyl,
halogen,
OH, NHz, CN, NOz, COOH, or SOz~.. In other embodiments, the substituted
carboxylic group can be substituted with a moiety selected from the group
consisting
of C1_lo alkyl, CN, COOH, NOz, NHz, SO2_4, C1-zo heteroalkyl, Cz_zo alkenyl,
alkynyl,
akynyl-aryl, alkynyl-heteroaryl, aryl, C1_zo alkyl-aryl, Cz_zo alkenyl-aryl,
heteroaryl,
C1-zo alkyl-heteroaryl, Cz_zo alkenyl-heteroaryl, cycloalkyl,
heterocycloalkyl, C1_zo
alkyl-heteroycloalkyl, and Cl_zo alkyl-cycloalkyl, any of which may be,
optionally,
substituted with a moiety selected from the group consisting of Cl_6 alkyl,
halogen,
OH, NHz, CN, NOz, COOH, or SOz_4. Exemplary heterocyclic groups include, but
are not limited to, morpholine, triazole, imidazole, pyrrolidine, piperidine,
piperazine,
pyrrole, dihydropyridine, aziridine, thiazolidine, thiazoline,
thiadiazolidine, and
thiadiazoline.
In specific embodiments, X can be hydroxyl, hydroxycarbonyl, 1-methyl-1-
cyclohexylcarbonyloxy, 1-methyl-1-cyclohexylmethoxycarbonyl, 5-ethyl-2-pyridyl-
acetoxy, 5-ethyl-2-pyridylmeth-oxy-carbonyl, (R)-6-hydroxy-2,5,7,8-tetramethyl-
chroman-2-carboxy, (S)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxy, (R)-6-
hydroxy-2,5,7,8-tetra-methylchroman-2-ylmethoxy -carbonyl, (S)-6-hydroxy-
2,5,7,8-
tetramethylchroman-2-ylmethoxycarbonyl, (R)-5-hydroxy-2,2,4,6,7-pentamethyl-
2,3-
dihydrobenzofuran-3-carboxy, (S)-5-hydroxy-2,2,4,6,7-pentamethyl-2,3-dihydro-
benzofuran-3-carboxy, (R)-5-hydroxy-2,2,4,6,7-penta-methyl-2,3-
dihydrobenzofuran-
3-methoxycarbonyl, (S)-5-hydroxy-2,2,4,6,7-pentamethyl -2,3-dihydrobenzofuran-
3-
methoxycarbonyl, 2-hydroxybenzoyloxy, or 2,4-dihydroxybenzoyloxy.
In other embodiments, X can be
OH OH O
Hetero O"'
wherein Hetero is an aromatic, cyclic, or alicyclic moiety that can contain
heteroatoms. In certain specific embodiments, Hetero is an aromatic, cyclic,
or
alicyclic moiety that contains heteroatoms that are generally part of the
structure of
the statin-family of lipid lowering agents. Preferred examples include, but
are not
limited to, 2-(4-fluorophenyl)-5-(1-methylethyl)-3-phenyl-4-
[(phenylamino)carbonyl]
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-1-(1H-pyrrol)yl, a component of atorvastatin, and 1,2,3,7,8,8a-hexahydro-1-(2-
methylbutanoyl)oxy-3,7-dimethyl-8-naphthalenyl, a component of lovastatin.
Alternatively, X can be
O
Fib
. . O
wherein Fib is an aromatic, cyclic, or alicyclic moiety that can contain
heteroatoms. In certain specific embodiments, Fib moieties are part of the
fibrate-
family of lipid lowering agents. Preferred examples include, but are not
limited to
4-(4-chlorobenzoyl)phenoxy, a component of fenofibric acid, 4-chlorophenoxy, a
component of clofibric acid, and 3-(2,5-xylyloxy)-1-propyl, a component of
gemfibrozil.
Alternatively, X can be
R
O
NSA
O
wherein R is hydrogen or methyl, and in which NSAID means an aromatic, alkyl,
or
cycloalkyl moiety that may contain heteroatoms and that are generally part of
the
family of non-steroidal anti-inflammatory agents. Preferred examples include,
but are
not limited to 4-(2-methyl-1-propyl)phenyl, 2-(2,6-dichloro-1-
phenyl)aminophenyl,
6'-methoxy-2'-naphthyl, and 6'-methoxy-2'-naphthylmethyl.
In another embodiment, X can be
i
E
s
where a and [3 are hydrogen or a, and (3 form a bond, and where y, 8, and s,
are
independently hydrogen, hydroxy, fluoro, chloro, or methyl.
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or
X can also be of the general formula
CH3
R2
N~~ R n
R1 O
O
In such embodiments, n is 0 or 1, R2 and R3 are independently hydrogen or
methyl; Z
is N, O, or S; and RI is aryl or heteroaryl, alkyl or heteroalkyl. Preferred
non-limiting
examples include compounds where Rl is phenyl, 4-fluorophenyl, 4-
methoxyphenyl,
3-methyl-2-thiophenyl, 5-methyl-2-thiophenyl, 5-methyl-3-isoxazolyl, 2-
pyridyl,
4-pyridyl, 2-pyrazinyl, 2-hydroxybenzoyl, or 2,4-dihydroxybenzoyl.
Other embodiments provide compounds wherein X is
CH3
R2
R ~N O n
O
Alternatively, X can be
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in which n is 0 or l, Ra and R3 are independently hydrogen or methyl; Z is N,
O, or S;
and Rl is aryl or heteroaryl, alkyl or heteroalkyl. Preferred non-limiting
examples
include compounds where Rl is phenyl, 4-fluorophenyl, 4-methoxyphenyl, 3-
methyl-
2-thiophenyl, 5-methyl-2-thiophenyl, 5-methyl-3-isoxazolyl, 2-pyridyl, 4-
pyridyl,
2-pyrazinyl, 2-hydroxybenzoyl, or 2,4-dihydroxybenzoyl.
In other embodiments, X is a 1-substituted (R)-pyrrolidine-2-
methoxycarbonyl, (S)-pyrrolidine-2-methoxycarbonyl, (R)-pyrrolidine-2-carboxy,
or
(S)-pyrrolidine-2-carboxy, having the following formulas
,N ,N ~N~
Y Y Y
O ~O O~O
O O "
in which Y is aryl or heteroaryl, alkyl or heteroalkyl. Preferred non-limiting
examples include compounds where Y is (R)-6-hydroxy-2,5,7,8-tetramethylchroman-
2-carboxy, (S)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxy, (R)-6-hydroxy-
2,5,7,8-tetrameth-ylchroman-2-ylmeth-oxycarbonyl, (S)-6-hydroxy-2,5,7,8-tetra-
methylchroman-2-ylmeth-oxycarbonyl, (R)-5-hydroxy-2,2,4,6,7-pentamethyl-2,3-
dihydrobenzofuran-3-carboxy, (S)-5-hydroxy-2,2,4,6 ,7-pentamethyl-2,3-dihydro-
benzofuran-3- carboxy, (R)-5-hydroxy-2,2,4,6,7-pentamethyl-2,3-
dihydrobenzofuran-
3-methoxycarbonyl, (S)-5-hydroxy-2,2,4, 6,7-pentamethyl-2,3-dihydrobenzofuran-
3-
methoxycarbonyl, 5-chloro-2-pyridyl, 5-methyl-2-pyridyl, 3-chloro-2-pyridyl,
4-methyl-2-pyridyl, 2-pyridyl, 2-benzoxazolyl, 2-benzothiazolyl, 5-amino-2-
pyridyl,
5-nitro-2-pyridyl, 2-pyrazinyl, 4-phenyl-2-oxazolinyl, 5-methyl-2-thiazolinyl,
4,5-dimethyl-2-oxazolinyl, 4,5-dimethyl-2-thiazolinyl, 5-phenyl-2-thiazolinyl,
2-thiazolinyl, 4-methyl-5-phenyl-2-thiazolinyl, 5-methyl-4-phenyl-2-
thiazolinyl,
2-piperidinyl, 4-phenyl-2-piperidinyl, 6-methyl-2-pyridinyl, 6-methoxy-2-
pyridinyl,
2-hydroxybenzoyl, or 2,4-dihydroxybenzoyl.
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Alternatively X is an N-substituted 2-methylaminoethoxycarbonyl or a
N-substituted 2-methylaminoacetoxy, having the following formulas:
CH3
CH3 Y-N~ O
Y -N
or
O O
O
in which Y is aryl or heteroaryl, alkyl or heteroalkyl. Preferred non-limiting
10 examples include compounds where Y is (R)-6-hydroxy-2,5,7,8-
tetramethylchroman-
2-carboxy, (S)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxy, (R)-6-hydroxy-
2,5,7,8-tetramethylchroman-2-ylmeth-oxycarbonyl, (S)-6-hydroxy-2,5,7,8-tetra-
methylchroman-2-ylmethoxycarbonyl, (R)-5-hydroxy-2,2,4,6,7-pentamethyl-2,3-
dihydrobenzofuran-3-carboxy, (S)-5-hydroxy-2,2,4,6, 7-pentamethyl-2,3-dihydro-
15 benzofuran-3-carboxy, (R)-5-hydroxy-2,2,4,6,7-pentamethyl-2,3-
dihydrobenzofuran-
3-methoxycarbonyl, (S)-5-hydroxy-2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran-3-
methoxycarbonyl, 5-chloro-2-pyridyl, 5-methyl-2-pyridyl, 3-chloro-2-pyridyl,
4-methyl-2-pyridyl, 2-pyridyl, 2-benzoxazolyl, 2-benzothiazolyl, 5-amino-2-
pyridyl,
5-nitro-2-pyridyl, 2-pyrazinyl, 4-phenyl-2-oxazolinyl, 5-methyl-2-thiazolinyl,
4,5-dimethyl-2-oxazolinyl, 4,5-dimethyl-2-thiazolinyl, 5-phenyl-2-thiazolinyl,
2-thiazolinyl, 4-methyl-5-phenyl-2-thiazolinyl, 5-methyl-4-phenyl-2-
thiazolinyl,
2-piperidinyl, 4-phenyl-2-piperidinyl, 6-methyl-2-pyridinyl, 6-methoxy-2-
pyridinyl,
2-hydroxybenzoyl, or 2,4-dihydroxybenzoyl.
X can also be a 1-substituted (R)-pyrrolidine-2-methoxycarbonyl,
(S)-pyrrolidine-2-methoxycarbonyl, (R)-pyrrolidine-2-carboxy, or (S)-
pyrrolidine-2-
carboxy, having the following formulas:
,N ,N ,N ~N~
y Y Y Y
O ~O O O O~O
O
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16
wherein Y is
CH3
Z R2
~ R3 n
/ 'N O
R1
n is 0 or 1; RZ and R3 are independently hydrogen or methyl; Z is N, O, or S;
and R1 is
aryl or heteroaryl, alkyl or heteroalkyl. Preferred non-limiting examples
include
compounds where Rl is phenyl, 4-fluorophenyl, 4-methoxyphenyl, 3-methyl-2-
thiophenyl, 5-methyl-2-thiophenyl, 5-methyl-3-isoxazolyl, 2-pyridyl, 4-
pyridyl, or 2-
pyrazinyl; or
Y is
~3 n
m
n is 0 or 1; m is 0 or 1; RZ and R3 are independently hydrogen or methyl; Z is
N, O, or
S; and Rl is aryl or heteroaryl, alkyl or heteroalkyl. Preferred non-limiting
examples
include compounds where Rl is phenyl, 4-fluorophenyl, 4-methoxyphenyl, 3-
methyl-
2-thiophenyl, 5-methyl-2-thiophenyl, 5-methyl-3-isoxazolyl, 2-pyridyl, 4-
pyridyl, or
2-pyrazinyl; or
Y is
OH OH O
Hetero
wherein Hetero is an aromatic, cyclic, or alicyclic moiety that usually
contains
heteroatoms. In certain specific embodiments, these moieties are part of the
structure
of the statin-family of lipid lowering agents. Preferred examples include, but
are not
limited to, 2-(4-fluorophenyl)-5-(1-methylethyl)-3-phenyl-4-
[(phenylamino)carbonyl]
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17
-1-(1H-pyrrol)yl, a component of atorvastatin, and 1,2,3,7,8,8a-hexahydro-1-(2-
methylbutanoyl)oxy-3,7-dimethyl-8-naphthalenyl, a component of lovastatin; or
Y is
O
Fib
wherein Fib is an aromatic, cyclic, or alicyclic moiety that contains
heteroatoms. In
some embodiments, these moieties are part of the fibrate-family of lipid
lowering
agents. Preferred examples include, but are not limited to 4-(4-
chlorobenzoyl)phenoxy, a component of fenofibric acid, 4-chlorophenoxy, a
component of clofibric acid, and 3-(2,5-xylyloxy)-1-propyl, a component of
gemfibrozil; or
Y is
R
NSAI ~ "
O
wherein R is hydrogen or methyl, and in which NSAID means an aromatic, alkyl,
or
cycloalkyl moiety that may contain heteroatoms and that are generally part of
the
family of non-steroidal anti-inflammatory agents. Preferred examples include,
but are
not limited to 4-(2-methyl-1-propyl)phenyl, 2-(2,6-dichloro-1-
phenyl)aminophenyl,
6'-methoxy-2'-naphthyl, and 6'-methoxy-2'-naphthylmethyl or
Y can be
i
3
E
s
where a and (3 are hydrogen or a and [3 form a bond, and where y, 8, and s,
are
independently hydrogen, hydroxy, fluoro, chloro, or methyl; or
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Y can be
0
or
Alternatively X can be an N-substituted 2-methylaminoethoxycarbonyl or an
N-substituted 2-methylaminoacetoxy, having the following formulas:
CH3
GH3 Y-N O
Y - N or
~ O O -
O-
wherein Y is
CH3
Z ~ R2
N R3 n
Rl O
n is 0 or 1; R2 and R3 are independently hydrogen or methyl; Z is N, O, or S;
and R1 is
aryl, heteroaryl, alkyl or heteroalkyl. Preferred non-limiting examples
include
compounds where Rl is phenyl, 4-fluorophenyl, 4-methoxyphenyl, 3-methyl-2-
thiophenyl, 5-methyl-2-thiophenyl, 5-methyl-3-isoxazolyl, 2-pyridyl, 4-
pyridyl, or
2-pyrazinyl, 2-hydroxybenzoyl, or 2,4-dihydroxybenzoyl; or
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Y is
n
m
n is 0 or 1; m is 0 or l; RZ and R3 are independently hydrogen or methyl; Z is
N, O, or
S; and Rl is aryl or heteroaryl, alkyl or heteroalkyl. Preferred non-limiting
examples
include compounds where Rl is phenyl, 4-fluorophenyl, 4-methoxyphenyl, 3-
methyl-
2-thiophenyl, 5-methyl-2-thiophenyl, 5-methyl-3-isoxazolyl, 2-pyridyl, 4-
pyridyl,
2-pyrazinyl, 2-hydroxybenzoyl, or 2,4-dihydroxybenzoyl; or
Y is
OH OH O
Hetero
wherein Hetero is an aromatic, cyclic, or alicyclic moiety that contains
heteroatoms.
In certain specific embodiments, these moieties are part of the structure of
the statin-
family of lipid lowering agents. Preferred examples include, but are not
limited to,
2-(4-fluorophenyl)-5-(1-methylethyl)-3-phenyl-4-[(phenylamino)carbonyl]-1-(1H-
pyrrol)yl, a component of atorvastatin, and 1,2,3,7,8,8a-hexahydro-1-(2-
methylbutanoyl)oxy-3,7-dimethyl-8-naphthalenyl, a component of lovastatin; or
Y is
O
Fib
wherein Fib is an aromatic, cyclic, or alicyclic moiety that contains
heteroatoms. In
some embodiments, these moieties are part of the fibrate-family of lipid
lowering
agents. Preferred examples include, but are not limited to 4-(4-
chlorobenzoyl)phenoxy, a component of fenofibric acid, 4-chlorophenoxy, a
component of clofibric acid, and 3-(2,5-xylyloxy)-1-propyl, a component of
gemfibrozil; or
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Y is
R
NSAI ~ ''
O
wherein R is hydrogen or methyl, and in which NSAID means an aromatic, alkyl,
or
cycloalkyl moiety that may contain heteroatoms and that are generally part of
the
5 family of non-steroidal anti-inflammatory agents. Preferred examples
include, but are
not limited to 4-(2-methyl-1-propyl)phenyl, 2-(2,6-dichloro-1-
phenyl)aminophenyl,
6'-methoxy-2'-naphthyl, and 6'-methoxy-2'-naphthyhnethyl; or
Y can be
i
i
s
s
10 where a and (3 are hydrogen or a, and (3 form a bond, and where y, b, and
s, are
independently hydrogen, hydroxy, fluoro, chloro, or methyl; or
Y can be
15 Other embodiments provide compounds wherein X is
O
O\ /
R ~ N O--
s
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R4 is hydrogen or methyl, and where RS is aryl or heteroaryl, alkyl or
heteroalkyl.
Preferred non-limiting examples include compounds where RS is phenyl,
4-fluorophenyl, 4-methoxyphenyl, 3-methyl-2-thiophenyl, 5-methyl-2-thiophenyl,
5-methyl-3-isoxazolyl, 2-pyridyl, 4-pyridyl, 2-pyrazinyl, (R)-6-hydroxy-
2,5,7,8-
tetramethyl-2-chromanyl, (S)-6-hydroxy-2,5,7,8-tetramethyl-2-chromanyl, (R)-5-
hydroxy-2,2,4,6,7-pentamethyl-2,3-dihydro-3-benzofuranyl, or (S)-5-hydroxy-
2,2,4,
6,7-pentamethyl-2,3-dihydro-3-benzo-furanyl.
X can also be
R / 'N O
5 \
O
wherein R4 is hydrogen or methyl, and where RS is aryl or heteroaryl, alkyl or
heteroalkyl. Preferred non-limiting examples include compounds where RS is
phenyl,
4-fluorophenyl, 4-methoxyphenyl, 3-methyl-2-thiophenyl, 5-methyl-2-thiophenyl,
5-methyl-3-isoxazolyl, 2-pyridyl, 4-pyridyl, 2-pyrazinyl, (R)-6-hydroxy-
2,5,7,8-
tetramethyl-2-chromanyl, (S)-6-hydroxy-2,5,7,8-tetramethyl-2-chromanyl, (R)-5-
hydroxy-2,2,4,6,7-pentamethyl-2,3-dihydro-3-benzofuranyl, or (S)-5-hydroxy-
2,2,4,
6,7-pentamethyl-2,3-dihydro-3-benzofuranyl.
In one embodiment, A is NH; B is sulfur (S); P and Q are a double bond or
hydrogen (H); E is hydrogen (H) and is attached to each of Di through D6; D1
through
D6 are carbon (C); and X can be any of the structures provided supra.
Also excluded, in certain optional embodiments, from the scope of the
presently claimed invention are compounds of Formula IB.
30
P
FORMULAIB Q O
D2 D3~ D4
B\ Fe A
6
X E
For compounds of Formula IB:
A, B, and F may be the same or different and are CH2, CO, N, NO, NH, SOo_a,
O;
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D1-D6 can be the same or different and are CH, N, S, or O;
E can be a substituent attached to one or more of the atoms located at D1-D6;
P and Q can be a double bond; or
P, Q, and E can be the same or different and are a moiety selected from the
group consisting of H, Cl_to alkyl, substituted alkyl groups, substituted or
unsubstituted carboxylic acids, substituted or unsubstituted carboxylic
esters, halogen,
carboxyl, hydroxyl, phosphate, phosphonate, aryl, CN, OH, COOH, NOz, NHz,
SOz_4,
Ci-zo heteroalkyl, Cz_zo alkenyl, alkynyl, akynyl-aryl, alkynyl-heteroaryl,
aryl, C1_zo
alkyl-aryl, Cz_zo alkenyl-aryl, heteroaryl, C1_zo alkyl-heteroaryl, Cz_zo
alkenyl-
heteroaryl, cycloalkyl, heterocycloalkyl, Cl_zo alkyl-heteroycloalkyl, and
C1_zo alkyl
cycloalkyl, any of which may be, optionally, substituted with a moiety
selected from
the group consisting of C1_6 alkyl, halogen, OH, NHz, CN, NOz, COOH, or SOz~.
Exemplary heterocyclic groups include, but not limited to, morpholine,
triazole,
imidazole, pyrrolidine, piperidine, piperazine, pyrrole, dihydropyridine,
aziridine,
thiazolidine, thiazoline, thiadiazolidine or thiadiazoline.
Substituted carboxylic acids, substituted carboxylic esters, and substituted
alkyl groups can be substituted at any available position with a moiety
selected from
the group consisting of CI_lo alkyl, halogen, CN, OH, COOH, NOz, NHz, SOz_4,
C1_zo
heteroalkyl, C2_zo alkenyl, alkynyl, akynyl-aryl, alkynyl-heteroaryl, aryl,
C1_zo alkyl-
aryl, Cz_zo alkenyl-aryl, heteroaryl, C1_zo alkyl-heteroaryl, Cz_zo alkenyl-
heteroaryl,
cycloalkyl, heterocycloalkyl, C1_zo alkyl-heteroycloalkyl, and C1_zo alkyl-
cycloalkyl,
any of which may be, optionally, substituted with a moiety selected from the
group
consisting of C1_6 alkyl, halogen, OH, NHz, CN, NOz, COOH, or 502. Exemplary
heterocyclic groups include, but are not limited to, morpholine, triazole,
imidazole,
pyrrolidine, piperidine, piperazine, pyrrole, dihydropyridine, aziridine,
thiazolidine,
thiazoline, thiadiazolidine, and thiadiazoline.
X is -OH, -COOH, or a substituted carboxylic group having the carboxyl
moiety OOC- or COO- directly attached to the phenyl ring of the compound of
Formula IB. The carboxylic acid group can be substituted with a moiety
selected
from the group consisting of alkyloxycarbonyl, alkylcarbonyloxy,
aryloxycarbonyl,
arylcarbonyloxy, heteroalkyloxycarbonyl, heteroalkylcarbonyloxy, heteroaryl-
oxycarbonyl, and heteroarylcarbonyloxy each of which is, optionally,
substituted with
Ci-to alkyl, CN, COOH, NOz, NHz, SOz~, C1_zo heteroalkyl, Cz_zo alkenyl,
alk5myl,
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23
akynyl-aryl, alkynyl-heteroaryl, aryl, C1_ZO alkyl-aryl, CZ_2o alkenyl-aryl,
heteroaryl,
Ci-ao alkyl-heteroaryl, CZ_2o alkenyl-heteroaryl, cycloalkyl,
heterocycloalkyl, C1_Zo
alkyl-heteroycloalkyl, and C1_ao alkyl-cycloalkyl, any of which may be,
optionally,
substituted with a moiety selected from the group consisting of C1_6 alkyl,
halogen,
OH, NH2, CN, NO2, COOH, or 502. In other embodiments, the substituted
carboxylic group can be substituted with a moiety selected from the group
consisting
of C1_lo alkyl, CN, COOH, N02, NH2, SO2~, Ci_2o heteroalkyl, C2_2o alkenyl,
alkynyl,
akynyl-aryl, alkynyl-heteroaryl, aryl, C1_ZO alkyl-aryl, CZ_ZO alkenyl-aryl,
heteroaryl,
Cl_ZO alkyl-heteroaryl, C2_2o alkenyl-heteroaryl, cycloalkyl,
heterocycloalkyl, C1_Zo
alkyl-heteroycloalkyl, and Cl_2o alkyl-cycloalkyl, any of which may be,
optionally,
substituted with a moiety selected from the group consisting of C1_6 alkyl,
halogen,
OH, NH2, CN, NO2, COON, or 502.x. Exemplary heterocyclic groups include, but
are not limited to, morpholine, triazole, imidazole, pyrrolidine, piperidine,
piperazine,
pyrrole, dihydropyridine, aziridine, thiazolidine, thiazoline,
thiadiazolidine, and
thiadiazoline.
In one exemplary embodiment, compounds of the invention of Formula IB
have the following moieties: A is NH; F is O; B is C=O; P and Q are a double
bond
or H; Dl-D6 are C (carbon), E is hydrogen; X is selected from the group
consisting of:
COOH, OH,
N
N~ N I ~N
% ~ ~ / O
O
O O
~ ~N N
\N N
> >
O
O
O
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CH3 O CH3
0
O O , and O O~ ; wherein
R1/\ N R2 Rl/\ N R2
N
R1= / ~ ~ ~ , or C ~ ; and
~ ' N N
RZ is CH3 or H.
The subject invention provides compounds comprising the following general
formula:
P Q
A~ R1
CH2~ R2
X-EY~ d ~- a
R4 R3
FORMULA II
wherein a is 0 to 4;
P and Q are H or CH3, or P and Q form a bond, therefore resulting in a double
bond between A and the adjacent carbon atom;
A is CH, N, O, or S; however, if A is O or S, then P is absent from Formula
II,
and Q is H or CH3;
Rl and R2 are linked and together form a chain having a length of 4- or 5-
atoms, said
chain containing at least 1 but optionally 2 or even 3 heteroatoms from the
group O,
S, or N, and said chain optionally containing at least 1 or 2 carbonyl (C=O)
groups;
or wherein Rl and RZ are not linked, and Rl can be -(C=0)NHZ, -(C=O)OH,
tetrazole, or -(C=O)0-C1_6 alkyl; and
R2 can be a hydrogen atom; Cl_3 alkyl; C1_6 alkoxy; Co_3 alkylenephenyl,
wherein the phenyl ring may be optionally substituted by 1 or more halogen
atoms;
tetrazole ring; (C=O)OH; (C=O)O-C1_6 alkyl;
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(C=O)bNR5R6, wherein b is 0 or 1; RS is H or C1_6 alkyl; and
R~ is H or B(C=O)~DR~ or B(CHOH)~DR~, where c is 0 or 1, B is a bond, a
C1_~ alkylene, a CZ_~ alkenylene, a C4_6 cycloalkenylene, a phenyl optionally
substituted by 1 or more C1_3 alkyl groups and/or 1 or more halogen atoms, or
a 5- or
5 6-membered heterocyclic group containing at least 1 or optionally 2
heteroatoms
(including any combination of O, N, or S at any position), where D is a bond,
a C1_3
alkyleneoxy, -O-, -NH-, or N(C1_3 alkyl)-, and where R~ is C1_6 alkyl, C4_6
cycloalkyl
or cycloalkenyl, phenyl optionally substituted by 1 or more halogen atoms,
Cl_3 alkyl,
CI_3 alkoxy, Co_3 alkyleneNR8R9 (each of R8 and R9 being independently H, C1_3
alkyl,
10 SOZC1_3alkyl, (C=O)OC1_3alkyl, S02NHC1_3alkyl), Co_3alkyleneCOOH, Co_
3alkylene(C=O)OC1_3alkyl, OCHZ(C=O)NH2, a 5- or 6-membered heterocyclic ring
containing at least 1 or optionally 2 heteroatoms (including any combination
of O, N,
or S at any position), or a fused bicyclic ring containing a benzene ring
fused with a 5-
or 6-membered heterocyclic ring containing at least 1 heteroatom (including O,
N, or
15 S at any position), and optionally substituted by an oxo (=O) group,
wherein said
bicyclic fused ring can be attached to D via a ring atom of the heterocyclic
ring either
directly or through a C1_6 alkylene ERIO where E is O, S, or NRII-, Rlo and
Rll being
independently H or CI_3 alkyl;
R3 and R4 are, optionally, the same or different and can be H, CH3, CF3,
20 OCH3, or a halogen atom;
dis0orl;
X can be a C3_8 cycloalkyl optionally substituted by a Cl_3 alkyl, Cl_3
alkoxy,
trifluoromethyl, hydroxy, cyano, (C=O)OC1_6alkyl, amino, alkylamino, or
dialkylamino; phenyl ring, optionally substituted by any combination of one or
more
25 halogen atoms, C1_6 alkyl, Cl_6 alkoxy, C1_6 fluoroalkoxy, nitrile, or
NR12Ri3 where
R12 and RI3 are independently H or C1_6 alkyl; 5- or 6-membered heterocyclic
ring
containing at least 1, or optionally 2, or more heteroatoms such as O, S, or
N, said
heterocyclic ring being optionally substituted by a C1_3 alkyl, C1_3 alkoxy,
trifluoromethyl, hydroxy, cyano, (C=O)OC1_6alkyl, amino, alkylamino, or
dialkylamino, provided that the heterocyclic ring may not be aromatic; fused
bicyclic
ring containing a phenyl ring fused with a 5- or 6-membered heterocyclic ring
containing at least 1, or optionally 2 or more heteroatoms such as O, N, or S,
wherein
both rings can be, optionally, independently substituted by C1_3 alkyl, Cl_3
alkoxy,
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26
trifluoromethyl, hydroxy, cyano, (C=O)OC1_6alkyl, amino, alkylamino, or
dialkylamino, and the heterocyclic ring may not be aromatic, provided that if
d = 0,
then the bicyclic ring X is attached to Z either directly via a ring atom of
the
heterocyclic ring of X, or through a sequence (CHa)tGg(CHZ)h(C=O);, wherein G
is O,
S, NH, or NC1_3alkyl, f is 0-6, g = 0 or 1, h = 0-6, and i = 0 or 1; or if d =
1, then the
bicyclic ring X is attached to Y either directly between a ring atom of the
heterocyclic
ring of X and a nitrogen atom of Y, or through a sequence (CH2)~g(CH2)h(C=O);,
where f, g, h, i, and G are defined as above;
Y is one of the following:
,....
..."...
N N N N
,N,~,I,~. , ,""
I
~ N ~. '~ N ~'/~
, or
in which the nitrogen atom is attached to X as defined above and in which the
2-
position of the pyrrolidine ring is attached to Z, either directly or through
a methylene
group;
Z is a group that can be enzymatically hydrolyzed or reduced, said enzymatic
reduction or hydrolysis results in the cleaving of Z into 2 molecular
fractions
including moieties such as -O(C=O)-, -(C=O)O-, -(C=O)S-, -S(C=O)-, -O(C=O)O-, -
S-S-, -O-P(=O)(OC1_6alkyl)O-, -P(=O)(OC1_6alkyl)O-, -N=N-, -(C=O)NH-, -
NH(C=O)-, -NHSO2-, -SO2NH-, -SO3-, -03S-, cholesteryl-O(C=O)O-, cholesteryl-
O(C=O)-, androstane 17[3-(C=O)- wherein the androtane group can contain 1-4
double bonds and can be optionally substituted by 1 or 2 oxo-groups, 1-4
halogen
atoms, 1-4 hydroxyl groups, or 1-4 methyl groups;
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alternatively, Z can also represent the following groups:
/ O OR14 Ris O
\ ~ / ~O O O
O O ~ j ~ k , or
O
OH OH ~'O
wherein j and k are integers from 0 to 4, and R14 and Rls independently
represent H or
C1_3 alkyl.
In some embodiments, X-Y-Z- together represent HO-, HO(C=O)-, H2N, or
HO3S-. Other embodiments provide compounds wherein the carbon center bearing Q
and R2 can be of the (S)-, (R)-, or (R,S)-configuration. Yet other embodiments
are
provided wherein all the possible asymmetrical carbon centers can be of the
(S)-, (R)-,
or (R,S)-configuration. Unsaturated moieties can be of the cis- or trans-
configuration.
A subgroup of compounds according to the present invention are represented
by the formula of Formula III. These compounds represent the
thiazolidinediones
subgroup:
O
NH
R3 R~ S
;' O
X(~dZ
FORMULA III.
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2~
Another subgroup is represented by Formula IV. These represent the
isoxazolidinediones subgroup:
O
FORMULA IV.
Another embodiment is depicted by Formula V. The compounds of this
subgroup represent the benzylmalonate subgroup (P and Q are H) and the
benzylidenemalonate subgroup (P and Q form a bond):
O
R3 / v
i
\ / N
X(~dZ O H
R3 / COOH
COOH
X(Y~dZ FORMULA V.
Yet another embodiment provides compounds having Formula VI. These
represent the 2-phenoxyisobutyric acid subgroup:
R4
R3 / O COOH
X(Y~dZ FORMULA VI.
Other embodiments provide compounds of Formula VII, which represents the
N-amyl phenylalanine subgroup in which Ar is phenyl or a 5- or 6-membered
heteroaryl group containing at least 1 atom selected from the group O, S, or
N:
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R3 / COOH
HN O
X~,.~dZ \
O Ar
FORMULA VII.
Formula VIII represents the N-aryl phenylalanine subgroup in which Ar is
phenyl or a 5- or 6-membered heteroaryl group containing at least 1 atom from
the
group O, S, or N:
R3 / COOH
HN
X(Y)dZ Ar
O
FORMULA VIII.
Another subgroup is depicted by Formulae IXA and IXB, the phenoxyacetic
acid subgroup, where the carboxylic acid moiety can be replaced by a tetrazole
ring:
O N~N
/ N
Z \ / O OH X YZ ~ ~ O N
d H
X-Y
R4 R3 R~t R3
FORMULA IXA FORMZJLA IXB
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wherein X(Y)dZ can be the following:
CHI
O
)H
O
O
O~O
O
O -N O~
\ /
10 ~CH3 H3C
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31
CH3 O ~, CH3
O ~ O O ~ O O
~N / ~ ~N
CH3 O ~, CH3 O
~ N O-S-O
I I ~>--~ o
S ~S
/ ~ /
F3C' F3C
N ~ N
~ \~-N ."", H I ~ \~-N
O O
O
O
N N
\ N I \ \ N
~ ~--
o ~ o
o ~ o
o Pso ~o
is ~o .,.
or
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32
O
N H
\ ~~ N O/
O
O
O
Compounds of Formula III can be conveniently prepared by the Knoevenagel
reaction between an aldehyde and thiazolidine-2,4-dione, using for example
sodium
acetate in acetic anhydride, or piperidine and benzoic acid in methylene
chloride as a
reaction medium. This is illustrated in Figure 29 and Figure 30.
Alternatively, the
same compounds can be prepared by the method described in Figure 31, in which
papa-anisidine undergoes a diazotation reaction with sodium nitrite and
hydrochloric
acid, the diazonium chloride salt undergoing in turn a radicalar reaction with
methyl
acrylate and then a cyclization reaction with thiourea, the product of which
is
hydrolyzed to the thiazolidinedione molecule.
Compounds of Formulae IV and V can be conveniently prepared according to
Figure 32 where for example p-methoxybenzaldehyde reacts in step (i) with
dimethyl
malonate in methanol with a catalytic amount of piperidinium benzoate, giving
the
benzylidene product. In step (ii), the benzylidene is hydrolyzed in
methanol/NaOH/water and then is acidified with dilute HCl to give the diacid.
The
diacid in turn reacts with thionyl chloride to give the acid chloride. In step
(iii), the
acid chloride is dissolved in dichloromethane and triethylamine. Hydroxylamine
hydrochloride is added under ice-cooling, giving the isoxazolidine compound.
In step
(iv), the methoxy-group is cleaved readily by boron trifluoride, yielding the
phenolic
product. Finally, in step (v), the benzylidene is reduced by magnesium powder
in
ethanol, giving dimethyl 4-methoxybenzylmalonate.
Compounds of Formula VI can be conveniently prepared from the reaction of
a phenol with acetone, chloroform, and sodium hydroxide, as shown in Figure
33.
Compounds of Formulae VII and VIII are tyrosine derivatives that are
substituted on the tyrosine nitrogen. They can be conveniently synthesized
from
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33
tyrosine methyl ester and 2-benzoylcyclohexanone followed by reduction with
10%
Pd/C as a catalyst (Figure 34), or from tyrosine methyl ester and 2-
benzoylcyclohexanecarbonyl chloride, followed by reduction with 10% Pd/C as a
catalyst (Figure 35).
The X-(Y)a-Z group can be synthesized according to procedures that have
been published elsewhere, for example in Chao et al. WO 01/00603 Al, in Henke
et
al., J. Med. CIZena. (1998) 41:5020-5036, in Collins et al., J. Med. CTzem.
(1998)
41:5037-5054, in Cobb et al., J. Med. Cl~em. (1998) 41:5055-5069, and in
Druzgala et
al. PCT/LTSO1/13131, each of which is hereby incorporated by reference in its
entirety. Some of these procedures are exemplified in Figures 36, 37, and 38.
Modifications of the compounds disclosed herein can readily be made by
those skilled in the art. Thus, analogs, derivatives, and salts of the
exemplified
compounds are within the scope of the subject invention. With a knowledge of
the
compounds of the subject invention, and their structures, skilled chemists can
use
known procedures to synthesize these compounds from available substrates.
As used in this application, the terms "analogs" and "derivatives" refer to
compounds which are substantially the same as another compound but which may
have been modified by, for example, adding additional side groups. The terms
"analogs" and "derivatives" as used in this application also may refer to
compounds
which are substantially the same as another compound but which have atomic or
molecular substitutions at certain locations in the compound.
Analogs or derivatives of the exemplified compounds can be readily prepared
using commonly known, standard reactions. These standard reactions include,
but are
not limited to, hydrogenation, methylation, acetylation, and acidification
reactions.
For example, new salts within the scope of the invention can be made by adding
mineral acids, e.g., HCI, HZSO4, etc., or strong organic acids, e.g., formic,
oxalic, etc.,
in appropriate amounts to form the acid addition salt of the parent compound
or its
derivative. Also, synthesis type reactions may be used pursuant to known
procedures
to add or modify various groups in the exemplified compounds to produce other
compounds within the scope of the invention.
The subj ect invention further provides methods of treating disorders, such as
diabetes, atherosclerosis, hypercholesterolemia, and hyperlipidemia,
comprising the
administration of a therapeutically effective amount of esterified
thiazolidinedione
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34
analogs to an individual in need of treatment. Thiazolidinedione based
compounds
include troglitazone (for example, REZULIN), pioglitazone, and rosiglitazone.
Accordingly, the subj ect invention provides esterified thiazolidinedione
analogs and
pharmaceutical compositions of these esterified compounds. The compounds and
compositions according to the invention can also be administered in
conjunction with
other therapeutic compounds, therapeutic regimens, compositions, and agents
suitable
for the treatment of disorders, such as diabetes, atherosclerosis,
hypercholesterolemia,
and hyperlipidemia. Thus, the invention includes combination therapies wherein
the
compounds and compositions of the invention are used in conjunction with other
therapeutic agents for the treatment of disorders, such as diabetes,
atherosclerosis,
hypercholesterolemia, and hyperlipidemia.
The compounds of this invention have therapeutic properties similar to those
of the unmodified parent compounds. Accordingly, dosage rates and routes of
administration of the disclosed compounds are similar to those already used in
the art
and known to the skilled artisan (see, for example, Physicians' Desk
Reference, 54th
Ed., Medical Economics Company, Montvale, NJ, 2000).
The compounds of the subject invention can be formulated according to
known methods for preparing pharmaceutically useful compositions. Formulations
are described in detail in a munber of sources that are well known and readily
available to those skilled in the art. For example, Remington's Pharmaceutical
Science by E.W. Martin describes formulations that can be used in connection
with
the subject invention. In general, the compositions of the subject invention
are
formulated such that an effective amount of the bioactive compounds) is
combined
with a suitable carrier in order to facilitate effective administration of the
composition.
In accordance with the subject invention, pharmaceutical compositions are
provided which comprise, as an active ingredient, an effective amount of one
or more
of the compounds of the invention and one or more non-toxic, pharmaceutically
acceptable carriers or diluents. Examples of such carriers for use in the
invention
include ethanol, dimethyl sulfoxide, glycerol, silica, alumina, starch, and
equivalent
carriers and diluents. Additional therapeutic agents suitable for the
treatment of
disorders such as diabetes, atherosclerosis, hypercholesterolemia, and hyper-
lipidemia
can also be incorporated into pharmaceutical agents according to the
invention.
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Further, acceptable Garners can be either solid or liquid. Solid form
preparations include powders, tablets, pills, capsules, cachets, suppositories
and
dispersible granules. A solid Garner can be one or more substances that may
act as
diluents, flavoring agents, solubilizers, lubricants, suspending agents,
binders,
5 preservatives, tablet disintegrating agents or encapsulating materials.
The disclosed pharmaceutical compositions may be subdivided into unit doses
containing appropriate quantities of the active component. The uiut dosage
form can
be a packaged preparation, such as packeted tablets, capsules, and powders in
paper or
plastic containers or in vials or ampoules. Also, the unit dosage can be a
liquid based
10 preparation or formulated to be incorporated into solid food products,
chewing gum,
or lozenge.
Adverse drug-drug interactions (DDI), elevation of liver function test (LFT)
values, and QT prolongation leading to torsades de pointes (TDP) are three
major
reasons why drug candidates fail to obtain FDA approval. All these causes are,
to
15 some extent metabolism-based. A drug that has two metabolic pathways, one
oxidative and one non-oxidative, built into its structure is highly desirable
in the
pharmaceutical industry. An alternate, non-oxidative metabolic pathway
provides the
treated subject with an alternative drug detoxification pathway (an escape
route) when
one of the oxidative metabolic pathways becomes saturated or non-functional.
While
20 a dual metabolic pathway is necessary in order to provide an escape
metabolic route,
other features are needed to obtain drugs that are safe regarding DDI, TDP,
and LFT
elevations.
In addition to having two metabolic pathways, the drug should have a rapid
metabolic clearance (short metabolic half life) so that blood levels of
unbound drug
25 do not rise to dangerous levels in cases of DDI at the protein level. Also,
if the
metabolic half life of the drug is too long, then the CYP450 system again
becomes the
main elimination pathway, thus defeating the original purpose of the design.
In order
to avoid high peak concentrations and rapidly declining blood levels when
administered, such a drug should also be administered using a delivery system
that
30 produces constant and controllable blood levels over time.
The subject invention also provides therapeutically useful and effective
compounds and compositions for the treatment of diabetes and a variety of
related
disorders, such as hyperlipidemia, and atherosclerosis. Various classes of
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36
compounds, useful for the treatment of diabetes and related disorders, that
can be
modified according to the concepts outlined herein include compounds such as
the
glitazones, thiazolidinediones, and isoxazolidinediones
The compounds of this invention have one or more of the following
characteristics or properties:
1. Compounds of the invention are metabolized both by CYP450 and by a
non-oxidative metabolic enzyme or system of enzymes;
2. Compounds of the invention have a short (up to four (4) hours) non-
oxidative metabolic half life;
IO 3. Oral bioavailability of the compounds is consistent with oral
administration using standard pharmaceutical oral formulations; however, the
compounds, and compositions thereof, can also be administered using any
delivery
system that produces constant and controllable blood levels over time;
4. Compounds according to the invention contain a hydrolysable bond that
can be cleaved non-oxidatively by hydrolytic enzymes;
5. Compounds of the invention can be made using standard techniques of
small-scale and large-scale chemical synthesis;
6. The primary metabolites) of compounds) of this invention results) from
the non-oxidative metabolism of the compound(s);
7. The primary metabolite(s), regardless of the solubility properties of the
parent drug, is, or are, soluble in water at physiological pH and have, as
compared to
the parent compound, a significantly reduced pharmacological activity;
8. The primary metabolite(s), regardless of the electrophysiological
properties of the parent drug, has, or have, negligible inhibitory activity at
the IKR
(HERG) channel at normal therapeutic concentration of the parent drug in
plasma
(e.g., the concentration of the metabolite must be at least five times higher
than the
normal therapeutic concentration of the parent compound before activity at the
II~R
channel is observed);
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37
9. Compounds of the invention, as well as the metabolites thereof, do not
cause metabolic DDI when co-administered with other drugs;
10. Compounds of the invention, as well as metabolites thereof, do not elevate
LFT values when administered alone.
In some embodiments, the subj ect invention provides compounds have any
two of the above-identified characteristics or properties. Other embodiments
provide
for compounds having at least any three of the above-identified properties or
characteristics. In another embodiment, the compounds, and compositions
thereof,
have any combination of at least four of the above-identified characteristics
or
properties. Another embodiment provides compounds have any combination of five
to 10 of the above-identified characteristics or properties. In a preferred
embodiment
the compounds of the invention have all ten characteristics or properties.
In various embodiments, the primary metabolites) of the inventive
compounds, regardless of the electrophysiological properties of the parent
drug, has,
or have, negligible inhibitory activity at the IKR (HERG) channel at normal
therapeutic concentrations of the drug in plasma. In other words, the
concentration of
the metabolite must be at least five times higher than the normal therapeutic
concentration of the parent compound before activity at the IKR chaimel is
observed.
Preferably, the concentration of the metabolite must be at least ten times
higher than
the normal therapeutic concentration of the parent compound before activity at
the
IKR channel is observed.
Compounds according to the invention are, primarily, metabolized by
endogenous hydrolytic enzymes via hydrolysable bonds engineered into their
structures. The primary metabolites resulting from this metabolic pathway are
water
soluble and do not have, or show a reduced incidence of, DDI when administered
with
other medications (drugs). Non-limiting examples of hydrolysable bonds that
can be
incorporated into compounds according to the invention include amide, ester,
carbonate, phosphate, sulfate, urea, urethane, glycoside, or other bonds that
can be
cleaved by hydrolases.
Additional modifications of the compounds disclosed herein can readily be
made by those skilled in the art. Thus, analogs, derivatives, and salts of the
exemplified compounds are within the scope of the subject invention. With a
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38
knowledge of the compounds of the subject invention skilled chemists can use
known
procedures to synthesize these compounds from available substrates. As used in
this
application, the terms "analogs" and "derivatives" refer to compounds which
are
substantially the same as another compound but which may have been modified
by,
for example, adding additional side groups. The terms "analogs" and
"derivatives" as
used in this application also may refer to compounds which are substantially
the same
as another compound but which have atomic or molecular substitutions at
certain
locations in the compound.
The subject invention further provides novel drugs that are dosed via drug
delivery systems that achieve slow release of the drug over an extended period
of
time. These delivery systems maintain constant drug levels in the target
tissue or
cells. Such drug delivery systems have been described, for example, in
Remington:
The Science and Practice of Pharmacy, 19th Ed., Mack Publishing Co., Easton,
PA,
1995, pp 1660-1675, which is hereby incorporated by reference in its entirety.
Drug
delivery systems can take the form of oral dosage forms, parenteral dosage
forms,
transdermal systems, and targeted delivery systems.
Oral sustained-release dosage forms are commonly based on systems in which
the release rate of drug is determined by its diffusion through a water-
insoluble
polymer. There are basically two types of diffusion devices, namely reservoir
devices,
in which the drug core is surrounded by a polymeric membrane, and matrix
devices,
in which dissolved or dispersed drug is distributed uniformly in an inert,
polymeric
matrix. In actual practice, however, many diffusion devices also rely on some
degree
of dissolution in order to govern the release rate.
Dissolution systems are based on the fact that drugs with slow dissolution
rates inherently produce sustained blood levels. Therefore, it is possible to
prepare
sustained-release formulations by decreasing the dissolution rate of highly
water-
soluble drugs. This can be carried out by preparing an appropriate salt or
other
derivative, by coating the drug with a slowly soluble material, or by
incorporating it
into a tablet with a slowly soluble Garner.
In actual practice, most of the dissolution systems fall into two categories:
encapsulated dissolution systems and matrix dissolution systems. Encapsulated
dissolution systems can be prepared either by coating particles or granules of
drug
with varying thicknesses of slowly soluble polymers or by micro-encapsulation,
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which can be accomplished by using phase separation, interfacial
polymerization, heat
fusion, or the solvent evaporation method. The coating materials may be
selected
from a wide variety of natural and synthetic polymers, depending on the drug
to be
coated and the release characteristics desired. Matrix dissolution devices are
prepared
by compressing the drug with a slowly soluble polymer carrier into a tablet
form.
In osmotic pressure-controlled drug-delivery systems, osmotic pressure is
utilized as the driving force to generate a constant release of drug.
Additionally, ion-
exchange resins can be used for controlling the rate of release of a drug,
which is
bound to the resin by prolonged contact of the resin with the drug solution.
Drug
release from this complex is dependent on the ionic environment within the
gastrointestinal tract and the properties of the resin.
Parenteral sustained-release dosage forms most commonly include
intramuscular injections, implants for subcutaneous tissues and various body
cavities,
and transdermal devices. Intramuscular injections can take the form of aqueous
solutions of the drug and a thickening agent which increases the viscosity of
the
medium, resulting in decreased molecular diffusion and localization of the
injected
volume. In this mamzer, the absorptive area is reduced and the rate of drug
release is
controlled. Alternatively, drugs can be complexed either with small molecules
such as
caffeine or procaine or with macromolecules, e.g., biopolymers such as
antibodies and
proteins or synthetic polymers, such as methylcellulose or
polyvinylpyrrolidone. In
the latter case, these formulations frequently take on the form of aqueous
suspensions.
Drugs which are appreciably lipophilic can be formulated as oil solutions or
oil
suspensions in which the release rate of the drug is determined by
partitioning of the
drug into the surrounding aqueous medium. The duration of action obtained from
oil
suspensions is generally longer than that from oil solutions, because the
suspended
drug particles must first dissolve in the oil phase before partitioning into
the aqueous
medium. Water-oil (W/O) emulsions, in which water droplets containing the drug
are
dispersed uniformly within an external oil phase, can also be used for
sustained
release. Similar results can be obtained from O/W (reverse) and multiple
emulsions.
Implantable devices based on biocompatible polymers allow for both a high
degree of control of the duration of drug activity and precision of dosing. In
these
devices, drug release can be controlled either by diffusion or by activation.
In
diffusion-type implants, the drug is encapsulated within a compartment that is
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enclosed by a rate-limiting polymeric membrane. The drug reservoir may contain
either drug particles or a dispersion (or a solution) of solid drug in a
liquid or a solid-
type dispersing medium. The polymeric membrane may be fabricated from a
homogeneous or a heterogeneous non-porous polymeric material or a microporous
or
5 semi-permeable membrane. The encapsulation of the drug reservoir inside the
polymeric membrane may be accomplished by molding, encapsulation,
microencapsulation or other techniques. Alternatively, the drug reservoir is
formed by
the homogeneous dispersion of drug particles throughout a lipophilic or
hydrophilic
polymer matrix. The dispersion of the drug particles in the polymer matrix may
be
10 accomplished by blending the drug with a viscous liquid polymer or a semi-
solid
polymer at room temperature, followed by crosslinking of the polymer, or by
mixing
of the drug particles with a melted polymer at an elevated temperature. It can
also be
fabricated by dissolving the drug particles and/or the polymer in an organic
solvent
followed by mixing and evaporation of the solvent in a mold at an elevated
15 temperature or under vacuum.
In microreservoir dissolution-controlled drug delivery, the drug reservoir,
which is a suspension of drug particles in an aqueous solution of a water-
miscible
polymer, forms a homogeneous dispersion of a multitude of discrete,
unleachable,
microscopic drug reservoirs in a polymer matrix. The microdispersion may be
20 generated by using a high-energy dispersing technique. Release of the drug
from this
type of drug delivery device follows either an interfacial partition or a
matrix
diffusion-controlled process.
In activation-type implants, the drug is released from the semi-permeable
reservoir in solution form at a controlled rate under an osmotic pressure
gradient.
25 Implantable drug-delivery devices can also be activated by vapor pressure,
magnetic
forces, ultrasound, or hydrolysis.
Transdermal systems for the controlled systemic delivery of drugs are based
on several technologies. In membrane-moderated systems, the drug reservoir is
totally
encapsulated in a shallow compartment molded from a drug-impermeable backing
30 and a rate-controlling microporous or non-porous polymeric membrane through
which the drug molecules are released. On the external surface of the
membrane, a
thin layer of drug-compatible, hypoallergenic adhesive polymer may be applied
to
achieve an intimate contact of the transdermal system with the skin. The rate
of drug
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release from this type of delivery system can be tailored by varying the
polymer
composition, permeability coefficient or thickness of the rate-limiting
membrane and
adhesive.
In adhesive diffusion-controlled systems, the drug reservoir is formulated by
directly dispersing the drug in an adhesive polymer and then spreading the
medicated
adhesive, by solvent casting, onto a flat sheet of drug-impermeable backing
membrane to form a thin drug reservoir layer. On top of the drug-reservoir
layer,
layers of non-medicated, rate controlling adhesive polymer of constant
thickness are
applied to produce an adhesive diffusion-controlled drug-delivery system.
In matrix dispersion systems, the drug reservoir is formed by homogeneously
dispersing the drug in a hydrophilic or lipophilic polymer matrix. The
medicated
polymer is then molded into a disc with a defined surface area and controlled
thickness. The disc is then glued to an occlusive baseplate in a compartment
fabricated from a drug-impermeable backing. The adhesive polymer is spread
along
the circumference to form a strip of adhesive rim around the medicated disc.
In
microreservoir systems, the drug reservoir is formed by first suspending the
drug
particles in an aqueous solution of a water-soluble polymer and then
dispersing
homogeneously, in a lipophilic polymer, by high-shear mechanical forces to
form a
large number of unleachable, microscopic spheres of drug reservoirs. This
thermodynamically unstable system is stabilized by crosslinking the polymer in
situ,
which produces a medicated polymer disk with a constant surface area and a
fixed
thickness.
Targeted delivery systems include, but are not limited to, colloidal systems
such _ as nanoparticles, microcapsules, nanocapsules, macromolecular
complexes,
polymeric beads, microspheres, and liposomes. Targeted delivery systems can
also
include resealed erythrocytes and other immunologically-based systems. The
latter
may include drug/antibody complexes, antibody-targeted enzymatically-activated
prodrug systems, and drugs linked covalently to antibodies.
The invention also provides methods of producing these compounds.
It is another aspect of this invention to provide protocols by which these
conditions can be tested. These protocols include in vitro and in vivo tests
that have
been designed to: 1) ensure that the novel compound is metabolized both by
CYP450
and by hydrolytic enzymes; 2) that the non-oxidative half life of the parent
drug is no
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more than a certain value when compared to an internal standard (in preferred
embodiments, less than about four hours); 3) that the primary metabolite of
the parent
drug is the result of non-oxidative metabolism; 4) that the primary metabolite
of the
parent drug (regardless of the solubility properties of the parent drug) is
water soluble;
5) that the primary metabolite of the parent drug (regardless of the
electrophysiological properties of the parent drug) has negligible inhibitory
properties
toward IKR channel at concentrations similar to therapeutic concentration of
the
parent drug; 6) that the novel compound (regardless of its properties) does
not cause
metabolic DDI when co-administered with other drugs; and 7) that the novel
compound does not cause hepatic toxicity in primary human hepatocytes.
A further aspect of the subject invention provides procedures for synthesizing
the therapeutic compounds of interest. An exemplary synthesis scheme is shown
in
Figures 22-28. In step 1, (3-benzyl aspartate is suspended in triethylamine
and acetic
anhydride is added slowly at 0°C with stirring. A catalytic amount
ofDMAP is then
added under ice-cooling. The mixture is stirred overnight at room temperature
and
then ice-water is added. The pH is brought up to 9.0 with I~.OH solution and
the
product is extracted with ethyl acetate, dried, and concentrated.
In step 2, the acetamide group and the benzyl ester are cleaved with 6N HCl at
reflux for 2 hours. The resulting amino acid is then isolated, dried, and then
dissolved
in a solution of thionyl chloride in methanol. After refluxing for 4 hours,
the resulting
methyl ester 3 is obtained.
In step 3, the amine compound 3 is suspended in dichloromethane and benzoyl
chloride and triethylamine are added under ice-cooling. After stirnng for 5
hours at
room temperature, the product is washed with sodium bicarbonate solution,
dried, and
evaporated to give the benzamide 4.
In step 4, the oxazole 5 is formed by dissolving compound 4 in anhydrous
ethyl acetate and treating with a catalytic amount of sulfuric acid for 3
hours at 90°C.
The product is isolated as usual.
In step 5, the carboxylic acid 6 is obtained by treating 5 with 1 equivalent
amount of lithium hydroxide in methanolJwater.
Steps 6 and 7 can be combined in a one-pot reaction as follows: Acetylacetane
7 (l.Smo1) is dissolved in 450m1 of glacial acetic acid and the solution is
cooled to
5°C. Sodium nitrite (l.Smol in 150m1 of Water) is added slowly so that
the
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temperature stays between 5 and 7°C. Keep stirnng for 4 hours at room
temperature
then add zinc powder (3mo1) portionwise under ice-cooling. Keep stirring at
room
temperature until the reaction is over and then collect the product 9 by
filtration. Dry
thoroughly.
Steps 8 and 9 proceed as described before. The amine 9 reacts with benzoyl
chloride in dichloromethane in the presence of triethylamine in order to give
the
benzamide 10. The oxazole 11 is then obtained by cyclization with a catalytic
amount
of sulfuric acid at reflux in anhydrous ethyl acetate.
In step 10, treating the ketone 11 with 1 equivalent of methyl magnesium
iodide in tetrahydrofuran at -40°C gives the tertiary alcohol 12.
In step 11, the ketone 11 is reduced to the secondary alcohol 13 with sodium
borohydride in methanol.
In step 12, p-methoxybenzaldehyde 14 reacts with dimethyl malonate in
methanol with a catalytic amount of piperidinium benzoate, giving the
benzylidene
product 15.
In step 13, the benzylidene 15 is hydrolyzed in methanol/NaOH/water and
then is acidified with dilute HCl to give the diacid. The diacid in turn
reacts with
thionyl chloride to give the acid chloride 16.
In step 14, the acid chloride 16 is dissolved in dichloromethane and
triethylamine. Hydroxylamine hydrochloride is added under ice-cooling, giving
the
isoxazolidine 17.
In step 15, the methoxy-group in compound 17 is cleaved readily by boron
tribromide, yielding the phenolic compound 18.
In step 16, the benzylidene compound 15 is reduced by magnesium powder in
ethanol, giving dimethyl 4-methoxybenzylmalonate 19.
In steps 17, 18, and 19, compound 19 undergoes a similar sequence of
reactions as in steps 13, 14, and 15, i.e., hydrolysis with
NaOH/methanol/water and
subsequent reaction with thionyl chloride to give the acid chloride 20.
Compound 20
in turn reacts with hydroxylamine hydrochloride in dichloromethane and
triethylamine to give 21. Finally, cleavage of the ether function with boron
tribromide yields the phenolic compound 22.
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In step 21, p-carboxybenzaldehyde 24 reacts with 2,4-isoxalolidinedione 25
(made from malonyl chloride and hydroxylamine, step 20) in THF in the presence
of
piperidinium benzoate to give the benzylidene 26.
In step 22, compound 26 is reduced with magnesium powder in ethanol to give
3-(4-carboxybenzyl)-isoxazolidine-2,4-dione 27.
In step 23, the carboxylic acid 26 reacts with the secondary alcohol 13 in
dichloromethane in the presence of 1 equivalent amount of
dicyclohexylcarbodiimide
(DCC) and 4-dimethylaminopyridine (DMAP), giving the ester 28.
The same reaction takes place in step 24 between compounds 27 and 13,
giving the ester 29.
Compounds 28 and 29 are among the group of preferred isoxazolidinedione
analogs that have' therapeutic properties against NIDDM and related diseases
in
mammals.
In step 25, the phenolic compound 18 reacts with the carboxylic acid 6 in
dichloromethane in the presence of 1 equivalent amount of
dicyclohexylcarbodiimide
(DCC) and 4-dimethylaminopyridine (DMAP), giving the ester 30.
The same reaction takes place in step 26 between compounds 22 and 6, giving
the ester 31.
Compounds 30 and 31 are among the group of preferred isoxazolidinedione
analogs that have therapeutic properties against NIDDM and related diseases in
mammals.
Ethyl acetoacetate 32 undergoes the same chemical treatment in steps 27 to 29
as acetylacetone 7 in steps 6 to 9 (Fig. 3). Thus, compound 32 in glacial
acetic acid
reacts with sodium nitrite, and the resulting oxime intermediate is not
isolated but is
reduced with zinc powder in acetic acid to give the amine 33. The amine is
then
coupled with benzoyl chloride in dichloromethane in the presence of
triethylamine.
The resulting benzamide 34 is then cyclized with a catalytic amount of
sulfuric acid in
refluxing ethyl acetate, giving the substituted oxazole 35.
In step 30, the ethyl carboxylate function of compound 35 is reduced with
lithium aluminum hydride in THF to give the primary alcohol 36 (an analog of
compounds 12 and 13 .
In step 31, the ethyl carboxylate function of compound 35 is hydrolyzed in 6N
HCl to give the carboxylic acid 37 (an analog of compound ~.
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Compounds 1 through 4 (of Table I) can be conveniently prepared by the
Knoevenagel reaction between an aldehyde and thiazolidine-2,4-dione, using for
example sodium acetate in acetic anhydride, or piperidine and benzoic acid in
methylene chloride as a reaction medium. This is illustrated in Figure 2 and
Figure 3.
5 Alternatively, compound 1 can be prepared by the method described in Figure
4. In
this reaction scheme, para-anisidine undergoes a diazotation reaction with
sodium
nitrite and hydrochloric acid. The diazonium chloride salt undergoing, in
turn, a
radicalar reaction with methyl acrylate and then a cyclization reaction with
thiourea,
the product of which is hydrolyzed to the thiazolidinedione molecule.
10 The compounds described in Table I (compounds 5 to 32) can all be made via
an esterification reaction between 1 or 2 and an appropriately substituted
carboxylic
acid, or between 3 or 4 and an appropriately substituted alcohol. The
esterification
reaction can be facilitated by the presence of a~ catalyst in the reaction
medium, such
as a small amount of concentrated sulfuric acid for example. Preferably,
especially if
15 the alpha-position to the carbonyl is an asymmetric center, an activated
functional
derivative of the carboxylic acid is made. Numerous functional derivatives of
carboxylic acids used in esterification reactions have been described in the
scientific
literature. The most commonly used activated functional derivatives are aryl
chlorides, anhydrides and mixed anhydrides, and activated esters. In one
aspect of
20 this invention dicyclohexyl carbodiimide (DCC) was used as an activating
agent
(Figure 5).
Compounds 33 to 104 are functionalized 5-methyloxazole and functionalized
5-methylthiazole derivatives. They all have various functional groups attached
to the
2-position (R1 in Tables II to V), and at the 4-position, which is the
enzymatically
25 labile link with the thiazolidine portion of the molecule. The
enzymatically labile link
is either an ester (COO-) or a reverse ester (00C-) and can be substituted
with 0, 1, or
2 methyl groups at the alpha-position from the oxazole or thiazole ring (R2
and R3 in
Tables II to V).
The synthesis of compounds 33 to 104 is described in general terms in Figures
30 7-10. Figure 6 describes the synthesis of the 4-oxazoleacetic acid and the
4
oxazoleethanol moiety starting from aspartic acid derivatives in which R2 and
R3 are
methyl or hydrogen. In a typical example, y-benzyl aspartate is acetylated and
then
decarboxylated to benzyl 3-acetamido-4-oxovalerate using acetic anhydride as
an
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46
acetylating agent followed by potassium hydroxide in order to obtain the
decarboxylated product. This in turn is transformed into methyl 3-amino-4-
oxovalerate using standard hydrolytic and esterification procedures, for
example
refluxing in dilute hydrochloric acid followed by reaction in thionyl chloride
and
methanol. The Rl group is then introduced by acylating the 3-amino group using
the
appropriate acyl or aroyl chloride. There is almost no limitation to the
nature of the
Rl group being introduced at this stage, as shown in Tables II to V where
various Rl
groups are described. Cyclization to an oxazole ring is then effected using
sulfuric
acid as a catalyst in ethyl acetate as a solvent. At this stage, ester
hydrolysis using
lithium hydroxide in methanol gives the desired 4-oxazoleacetic acid
derivatives,
whereas reduction of the ester with lithium aluminum hydride or reduction of
the acid
using diborane gives the 4-oxazoleethanol analogs.
Figure 7 describes the synthesis of the 4-oxazolecarboxylic acid and
4-oxazolemethanol groups. The synthesis starts from ethyl acetoacetate in
which a
2-amino-group is introduced via oxime formation followed by reduction with
zinc
powder. The synthesis then proceeds as before, where the Rl group is
introduced by
acylating the amino group, followed by cyclization with sulfuric acid in ethyl
acetate,
and finally ester cleavage or reduction to the alcohol.
Figure 8 shows how steric hindrance can be introduced under the form of
methyl groups on the 4-methanol moiety. Starting from pentane-2,4-dione,
following
the same synthetic sequence as in Figure 7 leads to the 4-acetyloxazole
compounds
which can be reduced by sodium borohydride to the 4-(1-ethyl)oxazole.
Alternatively, the compounds can be transformed by methylmagnesium iodide into
the tertiary alcohol analogs. In another embodiment, condensation of a
thioamide
with methyl 4-bromo-3-oxopentanoate gives methyl 4-thiazoleacetate, as
described in
Figure 9. Ester cleavage with lithium hydroxide or reduction with lithium
aluminum
hydride gives the corresponding acid or the alcohol, respectively.
Compounds 105 to 224 in Tables VI to XVII all have an amino acid or an
amino alcohol as part of their structure. Their synthesis is described in
Figures 10 to
18. Any amino acid can be used in the synthesis of compounds according to this
aspect of the invention. In certain embodiments, the amino acid group can be
either
proline or N-methyl glycine and the amino alcohol group is their alcohol
equivalent,
i.e., prolinol or N-methyl glycinol, respectively. As shown in Figures 10 to
13, the
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47
reaction of an alkyl chloride or a 2-heteroaryl chloride with proline,
prolinol,
N-methyl glycine, or N-methyl glycinol, in THF and triethylamine gives the
corresponding N-alkyl or N-heteroaryl adduct, respectively. When these adducts
are
carboxylic acids, such as in Figures 10 and 12, they react with 5-(4-
hydroxybenzyl)thiazolidine-2,4-dione in the presence of DCC and DMAP to give
compounds 105-108, 111, 112, 125-128, 131, 132, 185-188, 191, 192. Carboxylic
acid adducts react with 5-(4-hydroxybenzylidene)thiazolidine-2,4-dione in the
presence of DCC and DMAP to give compounds 115-118, 121, 122, 135-138, 141,
142, 195-198, 201, 202. When these adducts are alcohols, such as in Figures 11
and
13, they react with 5-(4-carboxybenzyl)thiazolidine-2,4-dione in the presence
of DCC
and DMAP to give compounds 145-148, 151, 152, 165-168, 171, 172, 205-208, 211,
212. Alcohol adducts react with 5-(4-carboxybenzylidene)thiazolidine-2,4-dione
in
the presence of DCC and DMAP to give compounds 155-158, 161, 162, 175-178,
181,182, 215-218, 221, 222.
Alternatively, the amino acid or amino alcohol group can be linked to another
group via an amide function, such as described in Figures 14 to 17. The
synthesis of
such compounds is straightforward. When the compounds contain an amino acid,
as
in Figures 14 and 16, the synthetic sequence is an amide bond formation, ester
deprotection, and ester formation.
As an illustrative example, (R)-Trolox~ is combined with L-proline methyl
ester, in the presence of DCC and DMAP in methylene chloride to form an amide
intermediate. The methyl ester of the proline group is then cleaved with
lithium
hydroxide in methanol, and the resulting carboxylic acid is combined with 5-(4-
hydroxybenzyl)thiazolidine-2,4-dione in 'DCC/DMAP/methylene chloride to give
compound 109. The (S)-isomer, compound 110, is made in a similar way. The same
kind of synthetic scheme leads to compounds 113,114, 119,120, 123,124,
129,130,
133,134,139,140,143,144,189,190,193,194,199, 200, 203, and 204.
When the compounds contain an amino alcohol, as in Figures 15 and 17, the
synthetic sequence is an amide bond formation, followed by an ester bond
formation.
As an illustrative example, (R)-Trolox~ is combined with L-prolinol in the
presence
of DCC and DMAP in methylene chloride to form an amide intermediate. The
resulting amide is combined with 5-(4-carboxybenzyl)thiazolidine-2,4-dione in
DCC/DMAP/methylene chloride to give compound 149. The (S)-isomer, compound
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150, is made in a similar way. The same kind of synthetic scheme leads to
compounds 153,154,159,160, 163,164,169,170,173,174,179,180,183,184, 209,
210, 213, 214, 219, 220, 223, and 224.
Compounds 225 to 242 (Table XVIII) are oxazoline-4-carboxylic acid types of
compounds. Their synthesis (Figure 18) starts from serine (RS=H) or from
threonine
(RS=CH3) benzyl ester. The ester is coupled with an alkyl or an arylcarboxylic
acid
using for example EDC as a coupling agent. The serine or threonine group then
cyclizes into an oxazoline upon treatment with thionyl chloride. Coupling with
5-(4
hydroxybenzyl)thiazolidine-2,4-dione using DCC/DMAP/methylene chloride gives
compounds 225 to 242.
Compounds 243 to 248 (Table XIX) are thiazolidinedione molecules where X
is a group containing a substituted 2-methyl-2-propionyl residue. Examples
include
the 2-methyl-2-(4-chlorophenoxy)propionyl moiety (clofibryl moiety), the 2-
methyl
2-[4-(4-chlorobenzoyl)phenoxy]propionyl moiety (fenofibryl moiety), and 2,2
dimethyl-5-(2,5-xylyloxy)valeryl moiety (gemfibrozilyl moiety).
Compounds 249 to 252 ~ (Table XX) are thiazolidinedione molecules where X
is a group containing a substituted (R,R)-3,5-dihydroxyheptanoyl residue.
Examples
include the ((3R, 8R)-2-(4-fluorophenyl)-5-(1-methylethyl)-3-phenyl-4-[(phenyl
amino)carbonyl] 1H-pyrrole-1-([3,8,dihydroxy)heptanoyl group (atorvastatin),
and the
1,2,3,7,8,8a-hexahydro-1-(2-methylbutanoyl)oxy-3,7-dimethylnaphthalenyl-8
[(3R,SR)-7-heptan]oyl group (lovastatin). The synthesis of these compounds
proceeds as in the examples of Table I, (i.e., by a simple esterification
procedure
between the lipid-lowering agent and compound 1 or compound 2).
Compounds 253 to 260 (Table XXI) are thiazolidinedione molecules where X
is a group containing an arylacetic acid residue, such as in molecules that
have non-
steroidal anti-inflammatory properties. In these examples, the X group is an
ibuprofen, ibufenac, naproxen, diclofenac, or nabumetone residue. The
synthesis of
these compounds is a simple ester formation reaction between the X group and
compound 1 (P and Q are hydrogen) or compound 2 (P and Q form a bond).
Compounds 261 to 268 (Table XXII) are thiazolidinedione molecules where X
is a group containing a cortienic acid residue, such as in molecules that have
glucocorticoid anti-inflammatory properties. In these examples, the X group is
a
cortienic acid, 1,2-dihydrocortienic acid, 6a, 9a-difluoro-1,2-
dihydrocortienic acid,
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49
and a 9a-fluoro-16a-methyl-1,2-dihydrocortienic acid residue. The synthesis of
these
compounds is a simple ester formation reaction between the X group and
compound 1
(P and Q are hydrogen) or compound 2 (P and Q form a bond). Cortienic acid,
one of
the many metabolites of hydrocortisone in man, can be synthetized from
hydrocortisone by oxidation with sodium periodate. The substituted cortienic
acid
analogs can be made in an identical manner from the corresponding substituted
glucocorticoids. This oxidation procedure is described in detail in [Druzgala
P.:
Novel Soft Anti-inflammatory Glucocorticoids for Topical Application. Ph.D.
Dissertation (1985), University of Florida, Gainesville, FL, hereby
incorporated by
reference in its entiretyJ.
Representative compounds were chosen and evaluated for activity on serum
glucose and insulin levels in non-insulin dependent diabetic mellitus
(I~TIDDM) KK-
AY male mice. Post-treatment data for each group were transferred to a
percentage of
pretreatment values and unpaired Student's t test was used for comparison
between
vehicle and test substance treated groups. Results show a significant
reduction of
both serum glucose and serum insulin relative to the vehicle control group.
Reduction
in serum glucose and serum insulin levels were comparable to the reduction
observed
in the troglitazone-treated animals. The results are presented in Table XXI
and in
Figures 19 and 20.
In certain embodiments, the subject invention specifically excludes those
compounds taught in International Application Nos. PCT/LTS00/18211 (filed June
30,
2000, and having International Publication No. WO 01/02377), PCT/USO1/29853
(filed September 21, 2001, and having International Publication No. WO
02/24689),
and PCT/LTSO1/13131 (filed April 24, 2001, and having International
Publication
No. WO 01/81328); EP 0528734 (Adir et Compagnie); EP 0549365 (Sankyo
Company Ltd.); EP 0848004 (Shionogi & Co., Ltd.); WO 97/32863 (Torii
Pharmaceutical Co.); Japanese Patent Nos. JP9165371 and JP9301963; I~letzien
et al.
(Molecular Pharmacology, 41(2)393-398, 1992); Unangst et al. (J. Medicinal
Chemistry, 37(2):322-328, 1994); and Sohda et al. (Chemical and Pharmaceutical
Bulletin, 30(10):3580-3600, 1982), each of which is hereby incorporated by
reference
in its entirety, including all figures and formulae).
Also excluded in various embodiments of the invention are those compounds
of Formula IA and IB where A is NH; B is sulfur; F is C=O; P and Q are a
double
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bond or hydrogen; E is hydrogen and is attached to each of D1 through D~ or E
is a
hydrogen atom, tef~t-butyl, phenyl, iso-propyl group attached to one, or both,
of DZ
and D~; D1 through D~ are carbon, and X is COON or OH. Thus, when A is NH; B
is
sulfur; F is C=O; P and Q are a double bond or hydrogen; E is hydrogen and is
5 attached to each of D1 through D6; and D1 through D6 are carbon, X cannot be
COOH
or OH. Alternatively, when E is a hydrogen atom or t-butyl group attached to
both of
DZ and D6, either 1) P and Q cannot be a bond; andlor 2) X cannot be OH.
Also excluded from the scope of the subj ect invention are 5-[4-[2-(3-
trifluoromethylphenyl)-2-(methoxy)ethylaminocarbonyloxy]benzyl]thiazolidin-2,4-
10 dione and 4-(2,4-dioxothiazolidin-5-ylidenemethyl)benzoic acid, 5-(4-
hydroxybenzyl)-3-triphenylmethyl-thiazolidine-2,4-dione, 5-(4-
acetoxybenzylidene)thiazolidine-2,4-dione, 5-(4-acetoxybenzyl)thiazolidine-2,4-
dione, 5-(4-acetoxybenzyl)-3-triphenylmethyl-thiaolidine-2,4-dione, 5-(4-
hydroxybenzyl)-3-triphenylmethyl-thiaolidine-2,4-dione, 5-(4-(N-3,4-
15 dichlorophenylcarbamoyl)benzylidene)thiazolidine-2,4-dione, 5-(4-(3,4-
dichlorophenoxycarboynyl)benzylidene)thiazolidine-2,4-dione, 5-(4-(3,4-
dichlorophenylacetoxy)benylidene)thiazolidine-2,4-dione, and 5-(4-(3,4-
dichlorobenzoyloxy)benylidene)thiazolidine-2,4-dione.
In certain other embodiments of the invention, compounds exemplified in
20 Figures 39-40, are excluded from the scope of the instant invention.
For Figure 39, substituent groups, and their definitions are set forth as
follows.
R is an optionally substituted aromatic hydrocarbon group, an optionally
substituted
alicyclic hydrocarbon group, an optionally substituted heterocyclic group, or
an
optionally substitutued condensed heterocyclic group. The aromatic hydrocarbon
25 group means phenyl, biphenylyl, naphthyl and the like. It may be an aralkyl
group
such as benzyl. Preferred is phenyl. The alicyclic hydrocarbon group means
alicyclic
hydrocarbon group having 3 to 7 carbon atoms, and is exemplified by
cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopropenyl, cyclobutenyl,
cyclobutadienyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl,
cyclohexadienyl,
30 cycloheptenyl, cycloheptadienyl and the like, with preference given to
alicyclic
hydrocarbon group having 5 to 7 carbon atoms. Specific examples thereof
include
cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclopentadienyl,
cyclohexenyl,
cyclohexadienyl, cycloheptenyl and cycloheptadienyl, with particular
preference
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given to cyclopentyl and cyclohexyl. The heterocyclic group is a 5- or 6-
membered
heterocycle having, as an atom constituting the ring, 1 to 3, preferably 1 or
2, hetero
atoms selected from nitrogen atom, oxygen atom and sulfur atom, besides carbon
atom, preferably an aromatic heterocycle. Specific examples thereof include
thienyl,
furyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl,
isoxazolyl,
oxadiazolyl, thiadiazolyl, triazolyl, pyridyl, pyrazinyl, pyrimidinyl,
pyridazinyl,
triazinyl, dithiazolyl, dioxolanyl, dithiolyl, pyrrolidinyl, dithiadiazinyl,
thiadiazinyl,
morpholinyl, oxazinyl, thiazinyl, piperazinyl, piperidinyl, pyranyl and
thiopyranyl,
with preference given to thienyl, furyl, pyrrolyl, imidazolyl, pyridyl and
pyrimidinyl,
and particular preference given to pyridyl, pyrimidinyl and imidazolyl. The
condensed heterocyclic group is a ring wherein 5- or 6-membered heterocycles
having, as an atom constituting the ring, 1 to 3, preferably 1 or 2, hetero
atoms
selected from nitrogen atom, oxygen atom and sulfur atom, besides carbon atom,
preferably aromatic heterocycles have been condensed, or a ring, wherein such
heterocycle, preferably an aromatic heterocycle, and a 4- to 6-membered
aromatic
hydrocarbon ring, preferably a benzene ring, have been condensed. Specific
examples
thereof include furoisoxazolyl, imidazothiazolyl, thienoisothiazolyl,
thienothiazolyl,
imidazopyrazolyl, cyclopentapyrazolyl, pyrrolopyrrolyl, cyclopentathienyl,
thienothienyl, oxadiazolopyrazinyl, benzofurazanyl, thiadiazolopyridinyl,
triazolothiazinyl, triazolopyrmidinyl, triazolopyridinyl, benzotriazolyl,
oxazolopyrimidinyl, oxazolopyridinyl, benzoxazolyl, thiazolopyridazinyl,
thiazolopyrimidinyl, benzisothiazolyl, benzothiazolyl, pyrazolotriazinyl,
pyrazolothiazinyl, imidazopyrazinyl, purinyl, pyrazolopyridazinyl,
pyrazolopyrimidinyl, imidazopyridinyl, pyranopyrazolyl, benzimidazolyl,
indazolyl,
benzoxathiolyl, benzodioxolyl, dithiolopyrimidinyl, benzodithiolyl,
indolidinyl,
indolyl, isoindolyl, furopyrimidinyl, furopyridinyl, benzofuranyl,
isobenzofuranyl,
thienopyrazinyl, thienopyrimidinyl, thienodioxinyl, thienopyridinyl,
benzothienyl,
isobenzothienyl, cyclopentaoxazinyl, cyclopentafuranyl, benzothiadiazinyl,
benzotriazinyl, pyridoxazinyl, benzoxazinyl, pyrimidothiazinyl,
benzothiazinyl,
pyrimidopyridazinyl, pyrimidopyrimidinyl, pyridopyridazinyl,
pyridopyrimidinyl,
cinnolinyl, quinazolinyl, quinoxalinyl, benzoxathiinyl, benzodioxinyl,
benzodithiinyl,
naphthyridinyl, isoquinolyl, quinolyl, benzopyranyl, benzothiopyranyl,
chromanyl,
isochromanyl, indolinyl and the like, with preference given to benzoxazolyl,
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52
benzisothiazolyl, benzothiazolyl, benzimidazolyl, indazolyl, benzoxathiolyl,
benzodioxolyl, benzodithiolyl, indolyl, isoindolyl, benzofuranyl,
isobenzofuranyl,
benzothienyl, isobenzothienyl, benzothiadiazinyl, benzotriazinyl,
benzoxazinyl,
benzothiazinyl, cinnolinyl, quinazolinyl, quinoxalinyl, benzoxathiinyl,
benzodioxinyl,
benzodithiinyl, isoquinolyl, quinolyl, benzopyranyl, benzothiopyranyl,
chromanyl,
isochromanyl and indolinyl, and particular preference given to indolyl,
isoindolyl,
benzofuranyl, isobenzofuranyl, benzothienyl, isobenzothienyl, isoquinolyl and
quinolyl. The lower alkyl is a linear or branched alkyl having 1 to 6 carbon
atoms,
such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-
butyl, pentyl,
isopentyl, neopentyl, tert-pentyl, hexyl, isohexyl, 3,3-dimethylbutyl, 2,2-
dimethylbutyl and the like, with preference given to alkyl having 1 to 4
carbon atoms
such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-
butyl, and
particular preference given to methyl. The optionally substituted means that
the
group may be substituted by 1 to 3 substituents which may be the same or
different.
Specific examples thereof include lower alkyl such as methyl, ethyl, propyl,
butyl,
tert-butyl and the like; lower alkoxy such as methoxy, ethoxy, propoxy,
butoxy, tert-
butoxy and the like; halogen atom; nitro; cyano; hydroxy; acyl (e.g., lower
alkanoyl
such as formyl, acetyl, propionyl, butyryl, isobutyryl and the like, amyl such
as
benzoyl, naphthoyl and the like, and the like); acyloxy (acyl moiety being as
defined
above) such as formyloxy, acetyloxy, propionyloxy, butyryloxy, isobutyryloxy,
benzoyloxy and the like; aralkyloxy such as benzyloxy, phenethyloxy,
phenylpropyloxy and the like; mercapto; lower alkylthio such as methylthio,
ethylthio, propylthio, butylthio, isobutylthio, tert-butylthio and the like;
amino; lower
alkylamino such as methylamino, ethylamino, propylamino, isopropylamino,
butylamino and the like; di(lower)alkylamino such as dimethylamino,
diethylamino,
dipropylamino, diisopropylamino, dibutylamino and the like; carboxy; lower
alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl,
isopropoxycarbonyl, butoxycarbonyl, tert-butoxycarbonyl and the like;
acylamino
(acyl moiety being as defined above); trifluoromethyl; phosphoryl; sulfonyl;
sulfonyloxy; carbamoyl; sulfamoyl; lower alkylphosphonamide such as
methylphosphonamide, ~ ethylphosphonamide, propylphosphonamide,
isopropylphosphonamide and the like; methylenedioxy; lower alkoxyphosphoryl
such
as methoxyphosphoryl, ethoxyphosphoryl, propoxyphosphoryl,
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isopropoxyphosphoryl and the like; lower alkylsulfonyl such as methylsulfonyl,
ethylsulfonyl, propylsulfonyl, butylsulfonyl, tent-butylsulfonyl and the like;
lower
alkylsulfonylamino such as methylsulfonylamino, ethylsulfonylamino,
propylsulfonylamino, butylsulfonylamino, tert-butyrylsulfonylamino and the
like; and
the like, with preference given to hydroxy, lower alkyl, lower alkoxy,
aralkyloxy,
mercapto, lower alkylthio, vitro, halogen atom, trifluoromethyl, amino,
di(lower)alkylamino, lower alkylamino, acyl, cyano, carbamoyl, acyloxy,
sulfonyl,
carboxy and lower alkoxycarbonyl, and particular preference given to hydroxy,
lower
alkyl and lower alkoxy. As used herein, by lower is meant that the number of
carbon
atoms is preferably 1 to 6, more preferably 1 to 4.
Also excluded, in certain embodiments of this invention, are compounds such
as those exemplified in Figures 40A and 40B. For such compounds, R is an
optionally substituted aromatic hydrocarbon; an optionally substituted
alicyclic
hydrocarbon; an optionally substituted heterocyclic group; an optionally
substituted
condensed heterocyclic group; or a group exemplified in Figure 40B, wherein Rl
is an
optionally substituted aromatic hydrocarbon, an optionally substituted
alicyclic
hydrocarbon, an optionally substituted heterocyclic group or an optionally
substituted
condensed heterocyclic group, Rz and R3 are the same or different and each is
a
hydrogen atom or a lower alkyl, and X is an oxygen atom, a sulfur atom or a
secondary amino; R4 is a hydrogen atom, a lower alkyl or a hydroxy; RS is a
lower
alkyl optionally substituted by hydroxy; and P and Q are each a hydrogen atom
or P
and Q together form a bond, and pharmaceutically acceptable salts thereof.
Also
excluded, in certain embodiments of the subject invention, are compounds
wherein R
is an optionally substituted phenyl, an optionally substituted 5- or 6-
membered
aromatic heterocyclic group having 1 or 2 hetero atoms selected from sulfur
atom,
oxygen atom and nitrogen atom, or an optionally substituted condensed aromatic
heterocyclic group wherein such aromatic heterocyclic ring and a benzene ring
are
condensed, and pharmaceutically acceptable salts thereof; or where R is a
phenyl, a 5-
or 6-membered aromatic heterocyclic group having one or two hetero atoms
selected
from sulfur atom, oxygen atom and nitrogen atom, or a condensed aromatic
heterocyclic group wherein such aromatic heterocyclic ring and a benzene ring
are
condensed, and pharmaceutically acceptable salts thereof; or where R is a
phenyl, or a
condensed aromatic heterocyclic group wherein a benzene ring and a 5- or 6-
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54
membered heterocyclic group having sulfur atom are condensed, and
pharmaceutically acceptable salts thereof; or where R is a phenyl, a
benzothienyl or 1
methyl-1-(2-pyridylthio)methyl, and pharmaceutically acceptable salts thereof;
or
where R is a phenyl, and pharmaceutically acceptable salts thereof; or where R
is the
structure set forth in Figure 40B, provided that X is a sulfur atom and
Rl is an optionally substituted phenyl or an optionally substituted 5- or 6-
membered aromatic heterocyclic group having one or two hetero atoms selected
from
sulfur atom, oxygen atom and nitrogen atom, and pharmaceutically acceptable
salts
thereof; or
Rl is a 5- or 6-membered aromatic heterocyclic group having one or two
hetero atoms selected from sulfur atom, oxygen atom and nitrogen atom, and
pharmaceutically acceptable salts thereof; or
Rl is a 5- or 6-membered aromatic heterocyclic group having nitrogen atom,
and pharmaceutically acceptable salts thereof; or
Rl is pyridyl, and pharmaceutically acceptable salts thereof.
In certain specific embodiments, isoxazolidinedione derivatives set forth in
Figure 40 and selected from the group of 4-[4-[2-(2-phenyl-5-methyl-4-
oxazolyl)ethoxy]benzyl]-3,5-isoxazolidinedione; 4-[4-[2-(2-phenyl-5-methyl-4-
oxazolyl)ethoxy]benzylidene]-3,5-isoxazolidine dione; 4-[4-[2-(2-benzothienyl-
5-
methyl-4-oxazolyl)ethoxy]benzyl]-3,5-isoxazolidinedione; 4-[4-[2-[5-methyl-[2-
(2-
pyridylthio)ethyl]-4=oxazolyl]ethoxy]benzyl]-3,5-isoxazolidinedione; and
pharmaceutically acceptable salts thereof are excluded from the scope of the
invention.
EXAMPLES
Example 1 - To (S)-2-pyrrolidinemethanol (3.96g) in THF (30m1) is added
2-chlorobenzoxazole (5.90g) also in THF (80m1) and then, dropwise,
triethylamine
(3.96g). Stir at 50°C for 4 hours. Cool to room temperature and filter
out the solid.
Evaporate the solvent and dissolve the crude product in 5m1 of methylene
chloride.
Pass through a silica plug (50g) in a fritted filter funnel, and elute with
methanol/methylene chloride (10:90), applying suction until the product has
been
collected. The yield of (S)-1-(2-benzoxazolyl)-2-hydroxymethylpyrrolidine is
8.2g.
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Example 2 - To (S)-2-pyrrolidinemethanol (3.96g) in THF (30m1) is added
2-chlorobenzothiazole (6.50g) also in THF (80m1) and then, dropwise,
triethylamine
(3.96g). Stir at 50°C for 4 hours. Cool to room temperature and filter
out the solid.
5 Evaporate the solvent and dissolve the crude product in 5m1 of methylene
chloride.
Pass through a silica plug (50g) in a fritted filter funnel, and elute with
methanol/methylene chloride (10:90), applying suction until the product has
been
collected. The yield of (S)-1-(2-benzothiazolyl)-2-hydroxymethylpyrrolidine is
8.8g.
10 Example 3 - To (R)-2-pyrrolidinemethanol (10.1g) in THF (50m1) is added 4,5-
dimethylthiazole (14.8g) also in THF (100m1) and then, dropwise, triethylamine
(10.1g). Stir at 50°C for 4 hours. Cool to room temperature and filter
out the solid.
Evaporate the solvent and dissolve the crude product in lOml of methylene
chloride.
Pass through a silica plug (100g) in a fritted filter funnel, and elute with
15 methanol/methylene chloride (10:90), applying suction until the product has
been
collected. The yield of (R)-1-(4,5-dimethyl-2-thiazolyl)-2-
hydroxymethylpyrrolidine
is 19.5g.
Example 4 - 2-chloropyridine (12g) and 2-(methylamino)ethanol (100m1) are
stirred
20 under nitrogen at 120°C for 18 hours. Cool to room temperature and
then pour into
iced water (250m1). Extract with ethyl acetate (2x200m1). Dry over sodium
sulfate.
Filter. Evaporate to dryness. The crude product is distilled in vacuo to give
10.3g of
N-methyl-N-(2-pyridinyl)-2-aminoethanol, boiling at 110°C/l.OmmHg.
25 Example 5 - A solution of 2-chlorobenzoxazole (15.3g) in THF (100m1) is
added
dropwise to an ice-cold solution of 2-(methylamino)ethanol (8.0g) and
triethylamine
(10.1g) also in THF (100m1). The mixture is stirred at room temperature for 4
hours
and the solid is filtered off. The solvent is evaporated and the residue is
dissolved in
methylene chloride and passed through a silica plug (100g), eluting with
30 methanol/methylene chloride (10:90) until the product has been collected.
The yield
of N-methyl-N-(2-benzoxazolyl)-2-aminoethanol is 15.7g.
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Example 6 - Thionyl chloride (2.5m1) was added dropwise to an ice-cold
solution of
(R)-6-hydroxy-2,5,7,8-tetramethylchroman-2-ylcarbinol (5.1g) in anhydrous
methylene chloride (SOmI). The solution was stirred at 0°C for 1 hour
and then at
room temperature for another period of 2 hours. Wash with saturated sodium
bicarbonate solution (2x25m1), then with brine (25m1), and then with water
(25m1).
Dry over sodium sulfate, filter, and evaporate to dryness. The crude product,
(R)-6-
hydroxy-2,5,7,8-tetramethylchroman-2-ylmethyl chloride (5.2g) is used as is in
the
next step.
Example 7 - Thionyl chloride (2.5m1) was added dropwise to an ice-cold
solution of
(S)-6-hydroxy-2,5,7,8-tetramethylchroman-2-ylcarbinol (5.1g) in anhydrous
methylene chloride (SOmI). The solution was stiired at 0°C for 1 hour
and then at
room temperature for another period of 2 hours. Wash with saturated sodium
bicarbonate solution (2x25m1), then with brine (25m1), and then with water
(25m1).
Dry over sodium sulfate, filter, and evaporate to dryness. The crude product,
(S)-6-
hydroxy-2,5,7,8-tetramethylchroman-2-ylmethyl chloride (5.0g) is used as is in
the
next step.
Example 8 - A mixture of (R)-6-hydroxy-2,5,7,8-tetramethylchroman-2-ylmethyl
chloride (8.43g), triethylamine (2.6g), and 2-(methylamino)ethanol (40m1) is
stirred at
120°C under nitrogen for 16 hours. Cool to room temperature and pour
into iced
water (100m1). Extract with ethyl acetate (3x100m1) and wash the combined
organic
extracts with brine (100m1). Dry over sodium sulfate. Filter. Evaporate to
dryness.
The product, (R)-2-[N-(6-hydroxy-2,5,7,8-tetramethylchroman-2-ylmethyl)-N-
methylamino]ethanol weighs 9.0g.
Example 9 - A mixture of (S)-6-hydroxy-2,5,7,8-tetramethylchroman-2-ylmethyl
chloride (8.43g), triethylamine (2.6g), and 2-(methylamino)ethanol (40m1) is
stirred at
120°C under nitrogen for 16 hours. Cool to room temperature and pour
into iced
water (100m1). Extract with ethyl acetate (3x100m1) and wash the combined
organic
extracts with brine (100m1). Dry over sodium sulfate. Filter. Evaporate to
dryness.
The product, (S)-2-[N-(6-hydroxy-2,5,7,8-tetramethylchroman-2-ylmethyl)-N-
methylamino]ethanol weighs 8.9g.
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Example 10 - A mixture of 2-chlorobenzoxazole (3.7g), (L)-proline methyl
ester,
hydrochloride salt (4.0g), and triethylamine (4.9g) in anhydrous THF (SOmI) is
stirred
at room temperature for 18 hours. The solid is filtered off and washed with
THF
(lOml). The solution is evaporated to dryness and the crude product is
dissolved in
methylene chloride (5m1) and passed through a plug of silica (50g), eluting
with ethyl
acetate/methylene chloride (10:90). The product, (L)-N-(2-benzoxazolyl)-
proline
methyl ester (5.0g) is a crystalline solid.
Example 11 - A mixture of 2-chlorobenzoxazole (3.7g), (D)-proline methyl
ester,
hydrochloride salt (4.0g), and triethylamine (4.9g) in anhydrous THF (SOml) is
stirred
at room temperature for 18 hours. The solid is filtered off and washed with
THF
(lOml). The solution is evaporated to dryness and the crude product is
dissolved in
methylene chloride (5m1) and passed through a plug of silica (50g), eluting
with ethyl
acetate/methylene chloride (10:90). The product, (D)-N-(2-benzoxazolyl)-
proline
methyl ester (5.5g) is a crystalline solid.
Example 12 - (L)-N-(2-benzoxazolyl)-proline methyl ester (5.0g) is suspended
in a
mixture consisting of methanol (SOmI), water (5m1), and lithium hydroxide
(0.5g).
Stir for 18 hours at room temperature. Acidify to pH 4.5 with citric acid.
Extract
with ethyl acetate (4x50m1). Dry over sodium sulfate, filter, and evaporate to
dryness.
The product, (L)-N-(2-benzoxazolyl)-proline (4.3g) is an off white solid.
Example 13 - A mixture of (L)-proline (4.6g), 2-chlorobenzoxazole (6.6g), and
triethylamine (4.45g) in anhydrous THF (100m1) is stirred at reflux
temperature for 18
hours. Cool down to room temperature, filter off the solid and wash it with a
THF
(lOml). Evaporate the solvent. Add ethyl acetate (SOmI) and then 1N sodium
hydroxide (SOml). Stir for 5 minutes. Keep the aqueous phase. Wash again with
ethyl acetate (SOml). Acidify with citric acid to pH 4.5. Isolate the
precipitate by
filtration. The aqueous filtrate is extracted with ethyl acetate (4x30m1). Dry
over
sodium sulfate. Filter. Evaporate to dryness. The solids are dried in vacuo at
35°C
for 18 hours. The first crop of product weighs 4.77g. The second crop weighs
3.26g.
The total amount of product, (L)-N-(2-benzoxazolyl)-proline, is 8.03g.
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Example 14 - A mixture of (D)-proline (4.6g), 2-chlorobenzoxazole (6.6g), and
triethylamine (4.45g) in anhydrous THF (100m1) is stirred at reflux
temperature for 18
hours. Cool down to room temperature, filter off the solid and wash it with a
THF
(lOml). Evaporate the solvent. Add ethyl acetate (SOmI) and then 1N sodium
hydroxide (SOml). Stir for 5 minutes. Keep the aqueous phase. Wash again with
ethyl acetate (SOmI). Acidify with citric acid to pH 4.5. Isolate the
precipitate by
filtration. The aqueous filtrate is extracted with ethyl acetate (4x30m1). Dry
over
sodium sulfate. Filter. Evaporate to dryness. The solids are dried in vacuo at
35°C
for 18 hours. The first crop of product weighs 4.93g. The second crop weighs
2.908.
The total amount of product, (L)-N-(2-benzoxazolyl)-proline, is 7.838.
Example 15 - A mixture of 4-hydroxybenzaldehyde (122.12g), 2,4-
thiazolidinedione
(117.13g), piperidine (5.11g), and benzoic acid (6.11g) in toluene (1,000m1),
is stirred
at 80°C for 16 hours. Cool to room temperature and filter off the
yellow solid. Wash
the solid with methylene chloride (3x100m1) and then with methanol/methylene
chloride (30:70) (2x100m1). Dry in vacuo at 35°C until constant weight.
The yield of
product, 5-(4-hydroxybenzylidene)-2,4-thiazolidinedione, is 217.8g.
Example 16 - To p-anisidine (25g) in acetone (400m1) at between 0 and
5°C, add
dropwise a solution of sodium nitrite (15.41g) in water (SOml) and 12N
hydrochloric
acid (SOml) from 2 different funnels over a 15-minute period. Stir for another
5
minutes at 0°C. Add methyl acrylate (104.9g) and then warm up the
solution to 35°C.
Transfer into a 2-L Erlenmeyer flask and stir vigorously. While stirring, add
copper(I) oxide (0.7g) in several portions. Keep stirnng for as long as
nitrogen gas
evolves from the solution, then stir for another 4 hours. Evaporate the
organic solvent
and dilute the aqueous residue with water (200m1). Extract with methylene
chloride
(200m1). Dry over sodium sulfate, filter, and evaporate to dryness. The
product,
methyl 2-chloro-3-(4-methoxyphenyl)propanoate, is a dark oil weighing 42.96g.
Example 17 - Methyl 2-chloro-3-(4-methoxyphenyl)propanoate (31.44g), thiourea
(16.89g), and anhydrous sodium acetate (11.24g) in 2-methoxyethanol (100m1) is
stirred at 100°C for 4 hours. Cool to room temperature and place the
flask at 4°C for
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16 hours. The pale yellow solid is filtered off and is washed with hexanes
(SOmI).
Stir for 30 minutes in ethyl acetate/water (100m1:10m1). Filter. Crystallize
from hot
ethanol (600m1). After leaving at 4°C for 16 hours, the crystals are
filtered off and
stirred at reflux for 8 hours in a mixture of 2-methoxyethanol (100m1) and 2N
hydrochloric acid (20m1). Evaporate the solvent. Add ethyl acetate (200m1) and
water (200m1). Keep the organic phase and wash again with water (200m1). Dry
over
sodium sulfate, filter, evaporate to dryness. The product, 5-(4-
methoxybenzyl)thiazolidine-2,4-dione (16.7g) is an oil that solidifies upon
standing.
Example 18 - To a solution of 5-(4-methoxybenzyl)thiazolidine-2,4-dione
(14.3g) in
anhydrous methylene chloride (100m1) cooled to -40°C, add a 1.0M
solution of boron
tribromide in methylene chloride (63m1). The solution is left to warm up to
23°C and
is then stirred for another 16 hours. Pour into iced water (700m1) and stir
for 15
minutes. Isolate the precipitate by filtration. Wash the product with water
(50m1) and
then with methylene chloride (SOml). The yield of 5-(4-
hydroxybenzyl)thiazolidine-
2,4-dione is 12.8g.
Example 19 - A mixture of methyl 4-formylbenzoate (164.16g), 2,4-
thiazolidinedione
(117.13g), piperidine (5.l 1g), and benzoic acid (6.11g) in toluene (1,000m1),
is stirred
at 80°C for 16 hours. Cool to room temperature and filter off the
yellow solid. Wash
the solid with methylene chloride (3x100m1) and then with methanol/methylene
chloride (30:70) (2x100m1). Dry in vacuo at 35°C until constant weight.
The yield of
product, 5-(4-carbomethoxybenzylidene)-2,4-thiazolidinedione, is 258.0g.
Example 20 - A suspension of 5-(4-carbomethoxybenzylidene)-2,4-
thiazolidinedione
(26.3g) and magnesium turnings (24g) in anhydrous methanol (300m1) is stirred
at
45°C for 8 hours. Acidify to pH 5.0 with 6N HCl and then extract with
methylene
chloride (2x250m1). Dry over sodium sulfate, filter, and evaporate to dryness.
The
crude product is chromatographed on silica gel (1,300g), eluting with
methanol/methylene chloride (02:98). The yield of 5-(4-carbomethoxybenzyl)-2,4-
thiazolidinedione is 15.2g.
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Example 21 - A suspension of 5-(4-carbomethoxybenzylidene)-2,4-
thiazolidinedione
(50g) in 6N HCl (200m1) is stirred at reflux for 4 hours. The mixture is
cooled to 4°C
and the product is filtered off. The product is then washed with water
(2x100m1) and
is dried in vacuo at 40°C. The yield of 5-(4-carboxybenzylidene)-2,4-
5 thiazolidinedione is 45g.
Example 22 - A suspension of 5-(4-carbomethoxybenzyl)-2,4-thiazolidinedione
(50g)
in 6N HCl (200m1) is stirred at reflux for 4 hours. The mixture is cooled to
4°C and
the product is filtered off. The product is then washed with water (2x100m1)
and is
10 dried in vacuo at 40°C. The yield of 5-(4-carboxybenzyl)-2,4-
thiazolidinedione is
44g.
Example 23 - (R)-6-Hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (9.2g)
and 5-(4-hydroxybenzyl)thiazolidine-2,4-dione (8.3g) are dissolved in
methylene
15 chloride (100m1) a.nd THF (50m1). To this add dicyclohexylcarbodiimide
(7.6g) and
DMAP (0.5g), and then stir for 4 hours at room temperature. The solid is
removed by
filtration and is washed with a small amount of THF (20m1). The solvent is
removed
and the solid residue is stirred with methylene chloride (100m1) and left at
4°C for 16
hours. The product is isolated by filtration and dried in vacuo at
23°C. The yield of
20 5-~4-[(R)-6-hydroxy-2,5,7,8-tetramethylchroman-2-
carboxy]benzyl}thiazolidine-2,4-
dione is 12.4g.
Example 24 - (S)-6-Hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (9.2g)
and 5-(4-hydroxybenzyl)thiazolidine-2,4-dione (8.3g) are dissolved in
methylene
25 chloride (100m1) and THF (SOml). To this add dicyclohexylcarbodiimide
(7.6g) and
DMAP (0.5g), and then stir for 4 hours at room temperature. The solid is
removed by
filtration and is washed with a small amount of THF (20m1). The solvent is
removed
and the solid residue is stirred with methylene chloride (100m1) and left at
4°C for 16
hours. The product is isolated by filtration and dried in vacuo at
23°C. The yield of
30 5-{4-[(S)-6-hydroxy-2,5,7,8-tetramethylchroman-2-
carboxy]benzyl}thiazolidine-2,4-
dione is 13.38.
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Example 25 - (R)-6-Hydroxy-2,5,7,8-tetramethylchroman-2-carbinol (1.9g) and 5-
(4-
carboxybenzyl)thiazolidine-2,4-dione (1.8g) are dissolved in methylene
chloride
(20m1) and THF (lOml). To this add dicyclohexylcarbodiimide (1.6g) and DMA.P
(0.1g), and then stir for 4 hours at room temperature. The solid is removed by
filtration and is washed with a small amount of THF (5m1). The solvent is
removed
and the solid residue is stirred with methylene chloride (20m1) and left at
4°C for 16
hours. The product is isolated by filtration and dried in vacuo at 23
°C. The yield of
5-~4-[(R)-6-hydroxy-2,5,7,8-tetramethylchroman-2-methoxy]benzyl)thiazolidine-
2,4-
dione is 2.54g.
Example 26 - (S)-6-Hydroxy-2,5,7,8-tetramethylchroman-2-carbinol (1.9g) and 5-
(4-
carboxybenzyl)thiazolidine-2,4-dione (1.8g) are dissolved in methylene
chloride
(20m1) and THF (lOml). To this add dicyclohexylcarbodiimide (1.6g) and DMAP
(0.1g), and then stir for 4 hours at room temperature. The solid is removed by
filtration and is washed with a small amount of THF (5m1). The solvent is
removed
and the solid residue is stirred with methylene chloride (20m1) and left at
4°C for 16
hours. The product is isolated by filtration and dried in vacuo at
23°C. The yield of
5-~4-[(S)-6-hydroxy-2,5,7,8-tetramethylchroman-2-methoxy]benzyl)thiazolidine-
2,4-
dione is 2.17g.
Example 27 - (R)-6-Hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (4.6g)
and 5-(4-hydroxybenzylidene)thiazolidine-2,4-dione (4.2g) are dissolved in
methylene chloride (SOmI) and THF (25m1). To this add dicyclohexylcarbodiimide
(3.8g) and DMAP (0.25g), and then stir for 4 hours at room temperature. The
solid is
removed by filtration and is washed with a small amount of THF (lOml). The
solvent
is removed and the solid residue is stirred with methylene chloride (SOmI) and
left at
4°C for 16 hours. The product is isolated by filtration and dried in
vacuo at 23°C.
The yield of 5-~4-[(R)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxy]benzyl-
idene]thiazolidine-2,4-dione is 5.9g.
Example 28 - (S)-6-Hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (4.6g)
and 5-(4-hydroxybenzylidene)thiazolidine-2,4-dione (4.2g) are dissolved in
methylene chloride (SOml) and THF (25m1). To this add dicyclohexylcarbodiimide
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(3.8g) and DMAP (0.25g), and then stir for 4 hours at room temperature. The
solid is
removed by filtration and is washed with a small amount of THF (10m1). The
solvent
is removed and the solid residue is stirred with methylene chloride (SOmI) and
left at
4°C for 16 hours. The product is isolated by filtration and dried in
vacuo at 23°C.
The yield of 5-~4-[(S)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxy]benzyl
idene}thiazolidine-2,4-dione is 6.2g.
Example 29 - (R)-6-Hydroxy-2,5,7,8-tetramethylchroman-2-carbinol (3.8g) and 5-
(4-
carboxybenzylidene)thiazolidine-2,4-dione (3.6g) are dissolved in methylene
chloride
(40m1) and THF (20m1). To this add dicyclohexylcarbodiimide (3.2g) and DMAP
(0.2g), and then stir for 4 hours at room temperature. The solid is removed by
filtration and is washed with a small amount of THF (lOml). The solvent is
removed
and the solid residue is stirred with methylene chloride (40m1) and left at
4°C for 16
hours. The product is isolated by filtration and dried in vacuo at 23
°C. The yield of
5- f 4-[(R)-6-hydroxy-2,5,7,8-tetramethylchroman-2-methoxy]benzylidene}
thiazoli
dine-2,4-dione is 5.4g.
Example 30 - (S)-6-Hydroxy-2,5,7,8-tetramethylchroman-2-carbinol (3.8g) and 5-
(4-
carboxybenzylidene)thiazolidine-2,4-dione (3.6g) are dissolved in methylene
chloride
(40m1) and THF (20m1). To this add dicyclohexylcarbodiimide (3.2g) and DMAP
(0.2g), and then stir for 4 hours at room temperature. The solid is removed by
filtration and is washed with a small amount of THF (lOml). The solvent is
removed
and the solid residue is stirred with methylene chloride (40m1) and left at
4°C for 16
hours. The product is isolated by filtration and dried in vacuo at 23
°C. The yield of
5- f 4-[(S)-6-hydroxy-2,5,7,8-tetramethylchroman-2-methoxy]benzylidene}
thiazoli-
dine-2,4-dione is 5.2g.
Example 31 - (L)-N-(2-benzoxazolyl)-proline (3.26g) and 5-(4-hydroxybenzyl)
thiazolidine-2,4-dione (3.11g) are suspended in methylene chloride (100m1).
Add
DCC (2.89g) and DMAP (0.12g) and stir at room temperature for 4 hours. Filter
and
purify on 114g of silica, eluting with methanol/methylene chloride (02:98).
The yield
of 5- f 4-[(S)-1-(2-benzoxazolyl)pyrrolidne-2-carboxy]benzyl} thiazolidine-2,4-
dione
is 4.55g.
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Example 32 - (L)-1-(2-benzoxazolyl)pyrrolidine-2-carbinol (3.26g) and 5-(4-
carboxy-
benzyl)thiazolidine-2,4-dione (3.25g) are suspended in methylene chloride
(100m1).
Add DCC (2.88g) and DMAP (0.12g) and stir at room temperature for 4 hours.
Filter
and purify on 132g of silica, eluting with methanol/methylene chloride
(02:98). The
yield of 5-{4-[(S)-1-(2-benzoxazolyl)pyrrolidinyl-2-methoxycarbonyl] benzyl~-
thiazolidine-2,4-dione is 4.68g.
Example 33 - (D)-1-(2-benzoxazolyl)pyrrolidine-2-carbinol (3.26g) and 5-(4-
carboxy-
benzylidene)thiazolidine-2,4-dione (3.35g) are suspended in methylene chloride
(100m1). Add DCC (2.91g) and DMAP (0.12g) and stir at room temperature for 4
hours. Filter and purify on 108g of silica, eluting with methanol/methylene
chloride
(02:98). The yield of 5-~4-[(R)-1-(2-benzoxazolyl)pyrrolidinyl-2-
methoxycarbonyl]benzylidene~-thiazolidine-2,4-dione is 4.32g.
Example 34 - (D)-1-(2-benzoxazolyl)pyrrolidine-2-carbinol (3.26g) and 5-(4-
carboxy-
benzyl)thiazolidine-2,4-dione (3.25g) are suspended in methylene chloride
(100m1).
Add DCC (2.93g) and DMAP (0.12g) and stir at room temperature for 4 hours.
Filter
and purify on 162g of silica, eluting with methanol/methylene chloride
(02:98). The
yield of 5- f 4-[(S)-1-(2-benzoxazolyl)pyrrolidinyl-2-methoxycarbonyl]benzyl)-
thiazolidine-2,4-dione is 4.77g.
Example 35 - Triethylamine (8.3m1) is added dropwise to a stirred cold
solution of
ethyl 2-aminoacetoacetate hydrochloride (5.4g) and 4-methoxybenzoyl chloride
(5.2g) in dichloromethane (100m1). After stirring for 3 hours, the solution is
washed
with water (100m1), dried over sodium sulfate, filtered, and evaporated to
dryness.
The crude product, ethyl 2-(4-methoxy)phenylaminoacetoacetate weighs 6.7g.
Example 36 - Ethyl 2-(4-methoxy)phenylaminoacetoacetate (5.9g) and phosphorus
oxychloride (SOmI) are stirred at 100C for 30 minutes. The phosphorus
oxychloride is
removed by evaporation, and the residue is diluted with aqueous sodium
bicarbonate
and extracted with methylene chloride. After drying over sodium sulfate, the
solution
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is evaporated and the product is crystallized from hexane, giving ethyl 5-
methyl-2-(4-
methoxy)phenyl-4-oxazolecarboxylate (4.5g).
Example 37 - A solution of benzoyl chloride (17g) in ethyl acetate (40m1) is
added
dropwise, with stirring, in an ice-cold mixture of L-serine methyl ester,
hydrochloride
(15.5g), water (100m1), sodium bicarbonate (21.8g), and ethyl acetate (100m1).
After
stirnng for 2 hours, the organic phase is separated, dried over sodium
sulfate, and
evaporated to give crystalline N-benzoyl-L-serine methyl ester (22.0g).
Example 38 - A stirred mixture of N-benzoyl-L-serine methyl ester (21.0g),
thionyl
chloride (21.0g), and methylene chloride (150m1) is stirred at reflux for 1
hour. The
solvent is evaporated and the residue is diluted with cold water. Neutralize
with
sodium bicarbonate, and extract with ethyl acetate. Purification on silica gel
(250g),
eluting with methanol:methylene chloride (01:99), yields methyl (S)-2-phenyl-2-
oxazoline-4-carboxylate (15.2g).
Example 39 - A solution of benzoyl chloride (17g) in ethyl acetate (40m1) is
added
dropwise, with stirring, in an ice-cold mixture of L-threonine methyl ester,
hydrochloride (16.5g), water (100m1), sodium bicarbonate (21.8g), and ethyl
acetate
(100m1). After stirring for 2 hours, the organic phase is separated, dried
over sodium
sulfate, and evaporated to give crystalline N-benzoyl-L-threonine methyl ester
(21.5g).
Example 40 - A stirred mixture of N-benzoyl-L-threonine methyl ester (21.0g),
thionyl chloride (21.0g), and methylene chloride (150m1) is stirred at reflux
for 1
hour. The solvent is evaporated and the residue is diluted with cold water.
Neutralize
with sodium bicarbonate, and extract with ethyl acetate. Purification on
silica gel
(250g), eluting with methanol:methylene chloride (01:99), yields methyl (R,S)-
2-
phenyl-2-oxazoline-5-methyl-4-carboxylate (14.8g).
Example 41 - Activity in NIDDM KK-AY male mice. Non-inslin dependent diabetic
mellitus male mice, weighing 50 +/- 5g (9-10 weeks of age) were used. These
animals exhibited hyperinsulinemia, hyperglycemia, and islet atrophy. The test
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compounds 105, 115, and 155, and the positive control compound troglitazone
were
suspended in a 1% carboxymethylcellulose preparation and were given orally at
a
dose of l Omg/kg, twice a day, for 5 consecutive days. Blood sampling was
performed
before the first dose and then 90 minutes after the last dose. Serum glucose
and
5 insulin levels were measured. Percent reduction of serum glucose and insulin
levels
relative to the pre-treatment values are shown in Table XX and figures 20 and
21.
Example 42 - CYP assays
A series of assays to test for activity of 5 principal drug metabolizing
10 enzymes, CYP1A4, CYP2C9, CYP2C19, CYP2D6, and CYP3A4, as well as other
CYP450 subfamilies, have been designed and are now commercially available
either
as ready-to-use kits or as contract work. Commercial sources for these assays
include
for example Gentest and MDS Panlabs. These assays can test for activity of the
enzyme toward metabolism of the test compound as well as testing for kinetic
15 modification (inhibition or activation) of the enzyme by the substrate.
These ira vitro
protocols use simple rapid, low cost methods to characterize aspects of drug
metabolism and typically require less than 1 mg of test material.
Example 43 - High Throughput Cytochrome P450 Inhibition Screen
20 The majority of drug-drug interactions are metabolism-based and of these,
most involve CYP450. For example, if a new chemical entity is a potent CYP450
inhibitor, it may inhibit the metabolism of a co-administered medication,
potentially
leading to adverse clinical events. The inhibition of human CYP1A2, CYP2C8,
CYP2C9, CYP2C19, CYP2D6, CYP3A4 and other isoforms are assessed using
25 microsomal preparations as enzyme sources and the fluorescence detection
method
described in the literature (Crespi, C.L., et al. (1997) Microtiter plate
assays for
inhibition of human, drug-metabolizing cytochromes P450. Anal. Biochem.
248:188-
190; Crespi, C.L., et al. (1999) Novel High throughput fluorescent cytochrome
P450
assays. Toxicol. Sci. 48, abstr. No.323; Favreau, L.V., et al. (1999) Improved
30 Reliability of the Rapid Microtiter Plate Assay Using Recombinant Enzyme in
Predicting CYP2D6 Inhibition in Human Liver Microsomes. Drug Metab. Dispos.
27:436-439). Tests are conducted in 96-well microtiter plates and may use the
following fluorescent CYP450 substrates: resorufm benzyl ether (BzRes), 3-
cyano-7-
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ethoxycoumarin (CEC), ethoxyresorufm (ER), 7-methoxy-4-trifluoromethylcoumarin
(MFC), 3-[2-(N,N-diethyl-N-methylamino)ethyl]-7-methoxy-4-methylcoumarin
(AMMC), 7-benzyloxyquinoline (BQ), dibenzyfluorescein (DBF) or 7-benzyloxy-4-
trifluoromethylcoumarin (BFC). Multiple CYP3A4 substrates are available to
assess
substrate dependence of ICSO values, activation and the complex inhibition
kinetics
associated with this enzyme (Korzekwa, K.R., et al. (1998). Evaluation of
atypical
cytochrome P450 kinetics with two-substrate models: evidence that multiple
substrates can simultaneously bind to the cytochrome P450 active sites.
Biochemistry., 37, 4137-4147; Crespi, C.L. (1999) Higher-throughput screening
with
human cytochromes P450. Curr. Op. Drug Discov. Dev.2: 15-19). Data are
reported
as ICSO values or percent inhibition when using only one or two concentrations
of test
compound.
Example 44 - Metabolic Stability
Metabolic stability influences both oral bioavailability and half life;
compounds of lugher metabolic stability are less controllable in their
pharmacokinetic parameters. This combination of characteristics, or
properties, leads
to potential DDI and liver toxicity. This test measures the metabolic
stability of the
compound in the presence of CYP450, in the presence of hydrolytic enzymes, and
in
the presence of both CYP450 and hydrolytic enzymes.
Stability in the pf~esence of CYP450: With CYP450 substrates of low and
moderate in vivo clearance, there is a good correlation between ira vitro
metabolic
stability and in vivo clearance (Houston, J.B. (1994) Utility of in vitro drug
metabolism data in predicting in vivo metabolic clearance). This test uses
pooled
liver microsomes, S9 (human and/or preclinical species) or microsomal
preparations
with appropriate positive and negative controls. Assessment of both phase-I
and
phase-II enzymatic metabolism is possible, and a standard set of substrate
concentrations and incubations may be used. Metabolism is measured by loss of
parent compound HPLC analysis with absorbance, fluorescence, radiometric or
mass
spectrometric detection can be used.
Stability in tlae presence of hydrolytic enzymes: Hydrolytic enzymes in liver
cytosol, plasma, or enzymatic mixes from commercial sources (human and/or
preclinical species) are used to assess the metabolic stability of the novel
compounds
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67
of the invention. Appropriate positive and negative controls, as well as a
standard set
of substrate concentrations, are added in order to correlate ire vitro
observations with
in vivo metabolic half life. Metabolism is measured by loss of parent
compound.
HPLC analysis with absorbance, fluorescence, radiometric or mass spectrometric
detection can also be used.
Stability in the presence of both CYP450 arad IZydf°olytic enzynt.es:
This test
uses pooled liver microsomes, S9 (human and/or preclinical species) or
microsomal
preparations with appropriate positive and negative controls, combined with
hydrolytic enzymes from commercial sources, plasma, or cytosol to assess
metabolic
stability. The test can also be performed in primary hepatocytes (human andJor
preclinical species) or in perfused liver (preclinical species). The use of
positive and
negative controls, as well as a standard set of substrates allow for
correlations
between in vitro observations and in vivo metabolic half life.
Example 45 - CYPlAl Induction Screening
Induction of CYP1A1 is indicative of ligand activation of the aryl
hydrocarbon (Ah) receptor, a process associated with induction of a variety of
phase-I
and phase-II enzymes (Swanson, H.I. (1993) The AH-receptor: genetics,
structure and
function. Pharmacogenetics 3:213-230). Many pharmaceutical companies choose to
avoid development of compounds suspected as Ah-receptor ligands. This test
uses a
human lymphoblastoid cell line containing native CYPlAl activity that is
elevated by
exposure to Ah receptor ligands. Assays are conducted in 96-well microtiter
plates
using an oversight incubation with the test substances, followed by addition
of
7-ethoxy-4-trifluoromethylcoumarin as substrate. Dibenz(a,h)anthracene is used
as a
positive control inducer. A concurrent control test for toxicity or CYP1A1
inhibition
is available using another cell line that constitutively expresses CYP1A1.
Example 46 - Cytochrome P450 Reaction Phenotyping
The number and identity of CYP450 enzymes responsible for the metabolism
of a drug affects population variability in metabolism. Reaction phenotyping
uses
either liver microsomes with selective inhibitors or a panel of cDNA-expressed
enzymes to provide a preliminary indication of the number and identity of
enzymes
involved in the metabolism of the substrate. The amount of each cDNA-expressed
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68
enzyme is chosen to be proportional to the activity of the same enzyme in
pooled
human liver microsomes. Protein concentration is standardized by the addition
of
control microsomes (without CYP450 enzymes). A standard set of substrate
concentrations and incubations is used and metabolism of the drug is measured
by
loss of parent compound. Alternatively, HPLC analysis with absorbance,
fluorescence, radiometric or mass spectrometric detection can be used.
Example 47 - Drug Permeability Measurement in Caco-2, LLC-PKl or MDCK Cell
Monolayers
Drug permeability through cell monolayers correlates well with intestinal
permeability and oral bioavailability. Several mammalian cell lines are
appropriate for
this measurement (Stewart, B.H., et al. (1995) Comparison of intestinal
permeabilities
determined in multiple in vitro and in situ models: relationship to absorption
in
humans. Pharm. Res. 12:693-699; Irvine, J.D., et al. (1999). MDCK (Madin-Darby
Canine Kidney) cells: A tool for membrane permeability screening. J. Pharm.
Sci.
88:28-33). Apical to basolateral diffusion is measured using a standard set of
time
points and drug concentrations. These systems can be adapted to a high
throughput
mode. Liquid chromatography/mass spectroscopy (LC/MS) analysis is also
available
for analysis of metabolites. Controls for membrane integrity and comparator
compounds are included and data are reported as apparent permeability (Papp)
or
percent flux under fixed conditions.
Example 48 - Human P-glycoprotein (PGP) Screen
An ATPase assay is used to determine if the compounds interact with the
xenobiotic transporter MDRl (PGP). ATP hydrolysis is required for drug efflux
by
PGP, and the ATPase assay measures the phosphate liberated from drug-
stimulated
ATP hydrolysis in human PGP membranes. The assay screens compounds in a high
throughput mode using single concentration determinations compared to the
ATPase
activity of a known PGP substrate. A more detailed approach by determining the
concentration-dependence and apparent kinetic parameters of the drug-
stimulated
ATPase activity, or inhibitory interaction with PGP can also be used.
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Example 49 - PGP-Mediated Drug Transport in Polarized Cell Monolayers
P-glycoprotein (PGP) is a member of the ABC transporter superfamily and is
expressed in the human intestine, liver and other tissues. Localized to the
cell
membrane, PGP functions as an ATP-dependent efflux pump, capable of
transporting
many structurally unrelated xenobiotics out of cells. Intestinal expression of
PGP may
affect the oral bioavailability of drug molecules that are substrates for this
transporter.
Compounds that are PGP substrates can be identified by direct measurement of
their
transport across polarized cell monolayers. Two-directional drug transport
(apical to
basolateral permeability, and basolateral to apical PGP-facilitated efflux)
can be
measured in LLC-PKl cells (expressing human PGP cDNA) and in corresponding
control cells. Caco-2 cells can also be used. Concentration-dependence is
analyzed
for saturation of PGP-mediated transport, and apparent kinetic parameters are
calculated. Test compounds can also be screened in a higher throughput mode
using
this model. LC/MS analysis is available. Controls for membrane integrity and
comparator compounds are included in the assay system.
Example 50 - Protein Binding
LC/MS analysis can be used to assess the affinity of the test compound for
immobilized human serum albumin (Tiller, P.R., et al. (1995) Immobilized human
serum albumin: Liquid chromatography/mass spectrometry as a method of
determining drug-protein binding. Rapid comm. mass spectrom. 9:261-263).
Appropriate low, medium and high binding positive control comparators are
included
in the test.
Example 51 - Metabolite Production
Milligram quantities of metabolites can be produced using microsomal
preparations or cell lines. These metabolites can be used as analytical
standards, an
aid in structural characterization, or as material for toxicity and efficacy
testing.
Example 52 - Effect on Herg Channel
This assay tests the effect of parent drugs and metabolites) on Herg channels
using either a cloned Herg channel expressed in stable human embryonic kidney
cells
(HEK), or Chinese hamster ovary cells (CHO) transiently expressing the
Herg/MiRP-
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1-encoded potassium channel. Whole cell experiments are carried out by means
of
the patch-clamp technique and performed in the voltage-clamp mode.
In the test using HEK cells, cells are depolarized from the holding potential
of
-80mV to voltages between -80 and +60 mV in lOmV increments for 4 seconds in
5 order to fully open and inactivate the channels. The voltage is then stepped
back to
-SOmV for 6 seconds in order to record the tail current. The current is also
recorded
in the presence of test compounds in order to evaluate a dose-response curve
of the
ability of a test compound to inhibit the Herg channel.
In the test involving CHO cells, the cells are clamped at a holding potential
of
10 -60mV in order to establish the whole-cell configuration. The cells are
then
depolarized to +40mV for 1 second and afterwards hyper-/depolarized to
potentials
between -120 and +20mV in 20mV increments for 300mSec in order to analyze the
tail currents. To investigate the effects of test compounds, the cells are
depolarized
for 300mSec to +40mV and then repolarized to -60mV at a rate of O.SmV/mSec,
15 followed by a 200-mSec test potential to -120mV. After 6 control
stimulations, the
extracellular solution is changed to a solution containing the test compound,
and 44
additional stimulations are then performed. The peaks of the outward currents
and
inward tail currents are analyzed.
Activity on HERD channel can also be assessed using a perfused hear
20 preparation, usually guinea pig hear or other small animal. In this assay
the hear is
paced and perfused with a'solution containing a known concentration of the
drug. A
concentration-response curve of the effects of drug on QT interval is then
recorded
and compared to a blank preparation in which the perfusate does not contain
the drug.
25 Example 53 - Toxicity in Hepatocyte Cell Culture
This test is performed in primary human and porcine hepatocyte cultures.
Toxicity is determined by the measurement of total protein synthesis by pulse-
labeling with [14C]leucine (Kostrubsky, V.E., et al. (1997) Effect of taxol on
cytochrome P450 3A and acetaminophen toxicity in cultured rat hepatocytes:
30 Comparison to dexamethasone. Toxicol., Appl. Pharmacol. 142:79-86), and by
reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
using a
protocol described by the manufacturer (Sigma Chemical Co., St. Louis, MO).
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Hepatocytes can be isolated from livers not used for whole organ transplants
or from
male Hanford miniature pigs.
It should be understood that the reaction schemes and embodiments described
herein are for illustrative purposes only and that various modifications or
changes in
light thereof will be suggested to persons skilled in the art and are to be
included
within the spirit and purview of this application and the scope of the
appended claims.
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Formula I
Table I.
Compound X P and Q*
number
1 H
HO---
2 db
3 O H
HO
db
O H
O~
\ CH3 db
O H
-N O~
H3C ~ ~ db
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73
Table I (continued).
Compound X P and Q*
number
CH3 O
H C 03C H
3 / O-
HO db
CH3
11 CH3 O ~ H
H3C / O = CH3
12 HO \ db
CH3
CH3
13 H3C O H
/
CH3
HO \ 'CH3
14 CH H O db
3
CH3
H3C O H
/
CH3
HO \ 'CH3
16 CH3 H ~O db
O
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74
Compound X P and Q*
number
17 O H
~O
1 g CH3 db
19 O~ H
O
2p CH3 db
21 - N O H
22 H3C O db
23 O " H
-N O
24 H C ~ ~ ~ db
3
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Compound
X
number P and Q*
CH3
25 H3C 03C O H
O
HO
26 db
CH3
27 CH3 ~O H
H3C / O - CH30
28 HO ~ db
CH3
CH3
29 H3C / O H
CH3
HO \ 'CH3
H
30 CH3 O db
O
CH3
31 H3C / O H
CH3
HO \ 'CH3
32 CH3 H O db
O
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0
Z CH3
Table II. R,--(~ I I ~ S~ H
N O
Formula II R2 R' o
Compound
Z R1 R2 R3
number
33 O ~ ~ H H
34 O ~ ~ CH3 H
35 O \ ~ CH3 CH3
36 S ~ ~ CH3 H
F
37 O ~ CH3 H
F
38 S ~ CH3 H
H3C0 /
39 O ~ CH3 H
H3C0 /
40 S ~ CH3 H
S
41 O I ~ CH3 H
CH3
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Compound Z R1 R2 R3
number
S
42 S I ~ CH3 H
CH3
H3C
S
43 O I / H H
H3C S
44 O I / CH3 H
H3C S
45 S I / H H
H3C
46 O ~ CH3 H
O,N
H3C
47 S ~ H H
O,N
48 O ~ ~ CH3 H
N ~'
49 O / I CH3 H
N
N
50 O ~ CH3 H
N
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0
Z CH3 \ U
O
RAW I ~~~~Y\~~NH
O /
Table III. R o
Compound
Z R1 R2 R3
number
51 O ~ I H H
52 O ~ ~ CH3 H
53 O \ ~ CH3 CH3
54 S ~ ~ CH3 H
F
55 O ~ CH3 H
F
56 S ~ CH3 H
H3C0 /
57 O ~ CH3 H
H3C0 /
58 S ~ CH3 H
S
59 O I ~ CH3 H
CH3
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Compound
Z R1 R2 R3
number
S
60 S I ~ CH3 H
CH3
H3C S
61 O I / H H
H3C S
62 O I / CH3 H
H3C S
63 S I / H H
H3C
64 O ~ CH3 H
O,N
H3C
65 S ~ H H
O~N
66 O ~ ~ CH3 H
N ~~.
67 O / I CH3 H
N
N
68 O ~ CH3 H
N
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0
CH3
Table IV. R ~Z ~ o ~ ~ S~ H
N
~Ry O p
Compound
Z R1 R2 R3
number
69 O ~ ~ H H
/
70 O ~ ~ CH3 H
71 O ~ ~ CH3 CH3
72 S \ ~ CH3 H
F
73 O ~ CH3 H
F
74 S ~ CH3 H
H3C0 /
75 O ~ CH3 H
\
H3C0 /
76 S ~ CH3 H
S
77 O I ~ CH3 H
CH3
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Compound
Z R1 R2 R3
number
S
78 S I ~ CH3 H
CH3
H3C S
79 O I / H H
H3C S
80 O I / CH3 H
H3C S
81 S I / H H
H3C
82 O ~ CH3 H
O, N
H3C
83 S ~ H H
O~N
84 O ~ ~ CH3 H
N
85 O / I CH3 H
N
N
86 O ~ ~ CH3 H
N
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S2
O
Z CH3
Table V. R'~~ ~ o ~ i ~ H
N
Rz Rs O O
Compound
Z R1 R2 R3
number
g7 O ~ ~ H H
88 O ~ ~ CH3 H
89 O ~ ~ CH3 CH3
90 S ~ ~ CH3 H
F
91 O ~ CH3 H
F
92 S ~ CH3 H
H3C0
93 O ~ ~ CH3 H
H3C0
94 S ~ ~ CH3 H
s
95 O I ~ CH3 H
CH3
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Compound
Z R1 R2 R3
number
S
96 S I ~ CH3 H
CH3
H3C S
97 O I / H H
H3C S
98 O I / CH3 H
H3C S
99 S I / H H
H3C
100 O / CH3 H
O , N.
H3C
101 S ~ H H
O, N.
102 O ( CH3 H
N
103 O / I CH3 H
N
N
104 O ~ CH3 H
N
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O
N _ I \ ~\NH
Y/ H O
Table VI.
O O
Compound
Y
number
/ N
105
\ O
/ N
106
\ S
107
N
H3C N
108
H3C S
CH3 O
H3C / O C
109
HO \
CH3
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Compound
Y
number
CH3 O
H3C / O - CH3
110
HO
CH3
CH3
H3C ~ O C
111
HO
CH3
CH3 ~~'~,.' ,
H3C / O - CH3
112
HO
CH3
CH3
H3C / O
113 I CH3
HO \ 'CH3
CH3 H
O
CH3
H3C / O
114 I CH3
HO \ 'CH3
CH3 H
O
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O
N I \ ~ \NH
Table VII. y' H=-' O
O O
Compound
Y
number
/ N
115
O
/ N
116
S
117
N ~'
H3C N
118
H3C S
CH3 O
H3C / ~3C
119
HO
CH3
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Compound
Y
number
CH3 O
H3C / O ' CH3
120
HO
CH3
CH3
H3C ~ O C
121
HO
CH3
CH3 ~'~' ,
H3C / O - CH3
122
HO
CH3
CH3
H3C / O
123 I CH3
HO \ 'CH3
CH3 H
O
CH3
H3C / O
124 ~ I CH3
HO \ 'CH3
CH3 H o,
O
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88
O
N I \ ~\NH
Y~ ~ / S
Table VIII. H ~--O
O O
Compound
Y
number
/ N
125
O
N
126
S
127
N ~'
HsC N
128
H3C S
CH3 O
H3C / O C
129
HO
CH3
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~9
Compound
Y
number
CH3 O .~,
H3C / O = CH3
130
HO
CH3
CH3
H3C ~ O C
131
HO
CH3
CH3
H3C / O = CH3
132
HO
CH3
CH3
H3C / O
133 ~ CH3
HO ~ 'CH3
CH3 H
O
CH3
H3C / O
134 ~ ~ CH3
HO ~ H CH3
CH3
O
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O
y N~ I \ s NH
H/.~-O /
Table IX. O O
Compound
Y
number
/ N
135
O
/ N
136
S
137
N
H3C N
138 ~ ~
H3C S
CH3 O
H3C / O C
139
HO
CH3
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Compound
Y
number
CH3 O ~',
H3C / O - CH3
140
HO
CH3
CH3
H3C / O C
141
HO
CH3
CH3
H3C / O - CH3
142
HO
CH3
CH3
H3C / O
143 I CH3
HO \ 'CH3
CH3 H
O
CH3
H3C / O
144 ~ CH3
HO \ 'CH3
CH3 H
O
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0
Table X. /~ ~ ~ ~ ~NH
O / S
' =~N
/ H
1' O
Compound
Y
number
/ N
145
\ O
/ N
146
\ S
147
N
H3C N
148
H3C S
CH3 O
H3C / O C
149
HO
CH3
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Compound
Y
number
CH3 O ,
H3 C / O - CH3
150
HO
CH3
CH3
H3C ~ O C
151
HO
CH3
CH3
H3C / O - CH3
152
HO
CH3
CH3
H3C / O
153 I CH3
HO \ 'CH3
CH3 H
O
CH3
H3C / O
154 I CH3
HO \ 'CH3
CH3 H
O
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94
O
\NH
p I / S
Table XI. N
/ H o O
Y
Compound
Y
number
/ N
155
O
/ N
156
S
157
N ~~.
H3C N
158
H3C S
CH3 O
H3C / O C
159
HO
CH3
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Compound
Y
number
CH3 O
H3C / O - CH3
160
HO
CH3
CH3
H3C ~ O C
161
HO
CH3
CH3 ~'~' .,
H3C / O °- CH3
162
HO
CH3
CH3
H3C / O
163 I CH3
HO \ 'CH3
CH3 H !
O
CH3
H3C / O
164 I CH3
HO \ 'CH3
CH3 H
O
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0
~NH
N O
1, H O O
Table XII.
Compound
Y
number
N
165
O
N
166
s
167
N
H3C N
168
H3C S
CH3 O
H3C ~ H03C
169
HO
CH3
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Compound
Y
number
CH3 O
H3C / O - CH3
170
HO
CH3
CH3
H3C / H03C
171
HO
CH3
CH3
H3C / O - CH3
172
HO
CH3
CH3
H3C / O
173 ~ CH3
HO \ 'CH3
CH3 H a
O
CH3
H3C / O
174 ~ CH3
HO \ 'CH3
CH3 H
O
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Table XIII. N~~.,","~O
1, H
Compound
Y
number
/ N
175
O
/ N
176
S
177
N ~'
H3C N
178
H3C S
CH3 O
HsC / O C
179
HO
CH3
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Compound
Y
number
CH3 O
H3C / O - CH3
180
HO
CH3
CH3
H3C ~ O C
181
HO
CH3
CH3 ., ,
H3C / O - CH3
182
HO
CH3
CH3
H3C / O
183 CH3
HO \ 'CH3
CH3 H i
O
CH3
H3C / O
184 I CH3
HO \ 'CH3
CH3 Oi
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100
O
H3C
N I ~ NH
Table XIV. y' . ~O / S
O \\O
Compound
Y
number
/ N
185
\ O
/ N
186
\ S
187
N ~'
H3C N
188
H3C S
CH3 O
H3C / OsC
189
HO \
CH3
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Compound
Y
number
CH3 O
H3C / O - CH3
190
HO
CH3
CH3
H3C / HO3C
191
HO
CH3
CH3
H3C / O - CH3
192
HO
CH3
CH3
H3C / O
193 ~ CH3
HO \ 'CH3
CH3 H
O
CH3
H3C / O
194 ( CH3
HO \ 'CH3
CH3 H
O
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O
H3C
N I ~ NH
Table XV. y O
O O
Compound
Y
number
/ N
195
O
/ N
196
S
197
N ~~.
H3C N
198
H3C S
CH3 O
H3C / HO3C
199
HO
CH3
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Compound
Y
number
CH3 O
H3C / O - CH3
200
HO
CH3
CH3
H3C / HO3C
201
HO
CH3
CH3
H3C / O °- CH3
202
HO
CH3
CH3
H3C / O
203 ~ CH3
HO \ 'CH3
CH3 H
O
CH3
H3C / O
204 I CH3
HO \ 'CH3
CH3 H
O
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H3C
Table XVI. N~
Y
Compound
Y
number
/ N
205
O
/ N
206
S
207
N ~~.
H3C N
208
H3C S
CH3 O
HsC '/ 03C
209
HO
CH3
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Compound
Y
number
CH3 O
H3C / O - CH3
210
HO
CH3
CH3
H3C ~ O C
211
HO
CH3
CH3
H3C / O - CH3
212
HO
CH3
CH3
H3C / O
213 I CH3
HO \ 'CH3
CH3 H
O
CH3
HsC / O
214 I CH3
HO \ 'CH3
CH3 O
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O
NH
H3C N~/O I / S
Table XVII. /
1, O O
Compound
Y
number
/ N
215
O
/ N
216
S
217
N
H3C N
218
HsC S
CH3 O
H3C '/ O C
219
HO
CH3
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Compound
Y
number
CH3 O
H3C / O - CH3
220
HO
CH3
CH3
H3C ~ 03C
221
HO
CH3
CH3
H3C / O - CH3
222
HO
CH3
CH3
H3C / O
223 ~ CH3
HO \ 'CH3
CH3 H
O
CH3
H3C / O
224 I CH3
HO \ 'CH3
CH3 H
O
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RS O O
O~~O , S NH
~~ N
Table XVIII.
Compound
R4 R5
number
225 ~ ~ H
/
226 ~ ~ CH3
F
227 ~ H
F
228 ~ CH3
H3C0 /
229 ~ H
H3C0 /
230 ~ CH3
S
231 I / H
CH3
S
232 I ~ CH3
CH3
HsC S
233 I H
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Compound
R4 RS
number
H3C S
234 I / CH3
H3C
235 ~ H
O~N
H3C
236 ~ CH3
O, N
237 ~ ~ H
N
238 ~ ~ CH3
N
239 / I H
N
240 / ~ CH3
N
N
241 ~ H
N
N
242 ~ CH3
N
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o ~ ~ ~O
Flb \ I s NH
'O
Table XIX. O
Compound Fib P and
Q*
number
O
243 C1 ~ / H
244 db
~
O
C1
245 H
246 O db
247 O H
248 db
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OH O
NH
Table XX. O
Compound Hetero P and Q*
number
249 ~ O H
H-N
- \ O
N . OH
OH
250 I ~ db
F
HO
251 ~ H
HO O
O O
252 db
",
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P
R 0
NSAID~O
S
~N
Table XXI //o
Compound NSAID P and Q*
number
253 H
/ w
H3C0 \ /
254 db
255
/
H3CO ~ /
256 db
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Table XXI (continued)
Compound NSAID P and Q*
number
257 H
25g ~ I db
259 ~ ~ ~' H
C1 Cl
260 ~ ~ db
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P
X ~ I S~NH
Table XXII
0
Compound X P and
Q*
number
261 H
0
OH
HO ,,,,.
262 " db
0
o
263 H
0
OH
HO ,,,..
264 ~ " db
0
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Table XXII (continued)
Compound X P and Q*
number
265 H
0
OH
HO ",..
266 F db
o/
/
F
o
267 H
0
HO ,,...OH
.",~
~
CH3
268 / " db
F
O/
/
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Table XXIII: Activity in NIDDM Mice.
Compound Serum Glucose (%) Serum Insulin
(%)
Vehicle 0 1
105 40 10
115 36 13
155 37 9
Troglitazone 35 15
