Note: Descriptions are shown in the official language in which they were submitted.
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pWAR.MACEUTICALLY ACCEPTABLE SALTS OF
HETEROCYCLIC COMPOUNDS
Field of the Invention
The present invention relates. to pharmaceutically acceptable salts of
compound of the general formula (I), their derivatives, their analogs, their
tautomeria forms, their stereoisomersa their folymorphs, their
pharmaceutically acceptable solvates and pharmaceutically acceptable
compositions containing them.
~ a
O
P
ORS p+
O M
The present invention also relates to a process for the preparation of the
above said pharmaceutically acceptable salts, their derivatives, their
analogs,
their tautorrieric forms, their stereoisomersa their polymorphs,
pharmiaceutically acceptable solvates, and pharmaceutical compositions
containing them.
. The compounds of the present invention lower plasma glucose,
triglycerides, lower total cholesterol (TC) and increase high density
lipoprotein (HDL) and decrease low density lipoprotein (LDL)~ which have a
beneficial effect on coronary heart disease and atherosclerosis.
The compounds of general formula (I) are useful in reducing body
weight arid for the treatment andlor prophylaxis of diseases such as
atherosclerosis, stroke, peripheral vascular diseases and related disorders.
These compounds are useful for the treatment of hyperglycemia,
hyperlipidemia, hypercholesterolemia, lowering of atherogenic lipoproteins,
VLI7L (very low density lipoprotein) and LI~L. The compounds of the present
invention can be used for the treatment of certain renal diseases including
CONFIRMATION COPY
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glomerulonephritis, glomerulosclerosis, nephrotic syndrome, hypertensive
nephrosclerosis and nephropathy. The compounds of general formula (I) are
also useful for the treatment and/or prophylaxis of type 2 diabetes, leptin
resistance, atherosclerosis, impaired glucose tolerance, disorders related to
syndrome X such as hypertension, obesity, insulin resistance, coronary heart
disease and other cardiovascular disorders. These compounds may also be
useful as aldose reductase inhibitors, for improving cognitive functions in
dementia, treating diabetic complications, disorders related to endothelial
cell
activation, psoriasis, polycystic ovarian syndrome (PCOS), inflammatory
bowel diseases, osteoporosis, myotoriic dystrophy, . pancreatitis,
arteriosclerosis, retinopathy, xanthoina, eating disorders, inflammation and
for
the treatment of cancer. The compounds of the 'p'resent invention are also
useful in the treatment and/or prophylaxis of the above said diseases in
combination/concomittant with one or more HMG CoA reductase inhibitors,
hypolipidemic/hypolipoproteinemic agents such as fabric acid derivatives,
nicotinic acid, cholestyramine, colestipol and probucol.
Background of Invention
Atherosclerosis and other peripheral vascular diseases effect the quality
of life of millions of ,people. Therefore, considerable attention has been .
directed towards understanding the etiology of hypercholesterolemia and
hyperlipiderizia and development of effective therapeutic strategies.
Hypercholesterolerilia has been defined as plasma cholesterol level that
exceeds arbitrarily defined value called "normal" level. Recently, it has been
accepted that "ideal" plasma levels of cholesterol are much below the
"normal" level of cholesterol in the general population and the risk of
coronary artery disease (CAD) increases as cholesterol level rises above the
"optimum" (or "ideal") value. There is clearly a definite cause and effect-
relationship between hypercholesterolemia and CAD, particularly for
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individuals with multiple risk factors. Most of the cholesterol is present in
the
esterified forms with various lipoproteins such as Low density lipoprotein
(LDL), Intermediate density lipoprotein (IDL), High density lipoprotein
(HDL) and partially as Very low density lipoprotein (VLDL). Studies clearly
indicate that there is an inverse correlationship between CAD and
atherosclerosis with serum HDL-cholesterol concentrations, (Stampfer et al.,
N. Engl. J llPed., 325 (19'91), 373-381) and the risk of CAD increases with
increasing levels of LDL and VLDL.
In CAD, generally "fatty streaks" in carotid, coronary and cerebral
arteries, are found which are primarily free and esterified cholesterol.
Miller et
al., (BY. Med. .7., 282 (1981), 1741 - 1744) have shown that increase in HDL-
particles may decrease the number of sites of stenosis in coronary arteries of
human, and high level of HDL-cholesterol may protect against the progression
of atherosclerosis. Picardo et al., A~tef°iosclerosis 6 (1986) 434 -
441 have
shown by in vitro experiment that HDL is capable of removing cholesterol
from cells. They suggest that HDL may deplete tissues of excess free
cholesterol and transfer it to liver (Macikinnon et al., J. Biol. them. 261
(1986), 2548 - 2552). Therefore, agents that increase HDL cholesterol would
have therapeutic significance for the treatment of hypercholesterolemia and
coronary heart diseases (CHD).
Obesity is a disease highly prevalent in affluent societies and in the
developing world and is a major cause of morbidity and mortality. It is a
state
of excess body fat accumulation. The causes of obesity are unclear. It is
believed to be of genetic origin or promoted by an interaction between the
genotype and environment. Irrespective of the cause, the result is fat
deposition due to imbalance between the energy intake versus energy
expenditure. Dieting, exercise and appetite suppression have been a part of
obesity treatment. There is a need for efficient therapy to fight this disease
since it may lead to coronary heart disease, diabetes, stroke, hyperlipidemia,
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gout, osteoarthritis, reduced fertility and many other psychological and
social
problems.
Diabetes and insulin resistance is yet another disease which severely
effects the quality of large population in the world. Insulin resistance is
the
S diminished ability of insulin to exert its biological action across a broad
range
of concentrations. In insulin resistance, the body secretes abnormally high
amounts of insulin to compensate for this defect; failing which, the plasma
glucose concentration inevitably rises and develops into diabetes. Among the
developed countries, diabetes mellitus is a common probleW Gild is associated
with a variety of abnormalities including obesity, hypertension,
hyperlipidemia (J. C'li~. IhvesB., 75 (1985) 809 - 817; N. Ehgl. J. lVled 317
(1987) 350-357; J. Clin. Endocrinol. Metab., 66 (1988) 580 e- 583; J. Clira.
Invest., 68 (1975) 957 - 969) and other renal complications (patent
publication
No. ~WO 95/21608). It is now increasingly being recognized that insulin
resistance and relative hyperinsulinemia have a contributory role in obesity,
hypertension, atherosclerosis and type 2 diabetes mellitus. The association of
insulin resistance with obesity, hypertension and angina ha's been described
as
a syndrome haviilg insulin resistance as the central pathogenic link-Syndrome-
X.
Hyperlipidemia is the primary cause for cardiovascular (CHID) and
other peripheral vasculax diseases. High risk of CVD is related to the higher
LDL (Low Density Lipoprotein) and VLDL (Very Low Density Lipoprotein)
seen in hyperlipidemia. Patients having glucose intolerance/insulin resistance
in addition to hyperlipidemia have higher risk of CVD. Numerous studies in
the past have shown that lowering of plasma triglycerides and total
cholesterol, in particular LDL and VLDL and increasing HDL cholesterol help
in preventing cardiovascular diseases.
Peroxisome proliferator activated receptors (PPAR) are members of the
nuclear receptor super family. The gamma (y) isoform of PPAR (PPARy) has
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been implicated in regulating differentiation of adipocytes (E~cdocrifaology,
135 (19'94) 798-800) and energy homeostasis (Cell, 83 (1995) 803-812),
whereas the alpha (a) isofonn of PPAR (PPARa) mediates fatty acid
oxidation (Trend. Ehdocrin. Metab., 4 (1993) 291-296) thereby resulting in
reduction of circulating free fatty acid in plasma (Current Biol. 5 (1995) 618
-
621). PPAI~a agonists have been found useful for the treatment of obesity
(WO 97/36579). It has been recently disclosed that coW pounds which are
agonists for both PPARa and PPARy are suggested to be useful for the
treatment of syndrome X (WO 97/25042). Similar effect between the insulin
sensitizes (PPARy agonist) and HMG C~A reductase inhibitor has been
observed which may be useful for the treatment of atherosclerosis and
xanthoma (EP 0 753 298).
It is known that PPARy plays an important role in adipocyte
differentiation (Cell, 87 (1996) 377-389). Ligand activation of PPAR is
sufficient to cause complete terminal differentiation (Cell, 79 (1994) 1147
1156) including cell cycle withdrawal. PPARy is consistently expressed in
certain cells and activation of this nuclear receptor with PPARy agonists
would stimulate the terminal differentiation of adipocyte precursors and cause
morphological and molecular changes characteristics of a more differentiated,
less malignant state (Molecular Cell, (1998), 465-470; CaYCifzoge~cesis,
(1998), 1949-53; Proc. Natl. Acad. Sci., 94 (1997) 237-241) and inhibition of
expression of prostate cancer tissue (Cancef~ Research 58 (1998) 3344-3352).
This would be useful in the treatment of certain types of cancer, which
express
PPARy and could lead to a quite nontoxic chemotherapy.
Leptin resistance is a condition wherein the target cells are unable to
respond to leptin signal. This may give rise to obesity due to excess food
intake and reduced energy expenditure and cause impaired glucose tolerance,
type 2 diabetes, cardiovascular diseases and such other interrelated
complications. Kallen et al (Proc. Natl. Acad. Sci. (1996) 93, 5793-5796) have
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reported that insulin sensitizers which perhaps due to the PPAR agonist
expression lower plasma leptin concentrations. However, it has been recently
disclosed that compounds having insulin sensitizing property also possess
leptin sensitization activity. They lower the circulating plasma leptin
concentrations by improving the target cell response to leptin (WO 98/02159).
In our WO publication 99/08501 we have disclosed and described the
novel compounds of the formula (II),
N R~ O
i (CH2)n-WAr R5
z R3 YR7
Rg0
X
R1 .
R N
where X represents O or S ; the groups R1, R2 and group R3 when attached
to the carbon atom, may be same or different and represent hydrogen, halogen,
hydroxy, vitro, cyano, formyl or optionally substituted groups selected from
alkyl, cycloalkyl, alkoxy, cycloalkoxy, aryl, aryloxy, aralkyl, aralkoxy,
heterocyclyl, heteroaryl, heteroaralkyl, heteroaryloxy, heteroaralkoxy, acyl,
acyloxy, hydroxyalkyl, amino, acylamino, alkylamino, arylamino,
aralkylamino, aminoalkyl, allcoxycarbonyl, aryloxycarbonyl,
aralkoxycarbonyl, alkoxyalkyl, aryloxyalkyl, aralkoxyalkyl, alkylthio,
thioalkyl, alkoxycarbonylamino, aryloxycarbonylamilio,
aralkoxycarbonylamino, carboxylic acid or its derivatives, or sulfonic acid or
its derivatives; R1, R2 along with the adjacent atoms to which they are
attached may also form a substituted or unsubstituted 5-6 meinbered cyclic
structure containing carbon atoms with one or more double bonds, which may
optionally contain one or more heteroatoms selected from oxygen, nitrogen
and sulfur; R3 when attached to nitrogen atom represents hydrogen, hydroxy,
formyl or optionally substituted groups selected _ from alkyl, cycloalkyl,
alkoxy, cycloalkoxy, aryl, aralkyl, heterocyclyl, heteroaryl, heteroaralkyl,
acyl,
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acyloxy, ' hydroxyalkyl, amino, acylamino, alkylamino, arylamino,
aralkylamino, aW inoalkyl, aryloxy, arallcoxy, heteroaryloxy, heteroaralkoXy,
alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, alkoxyalkyl,
aryloxyalkyl, aralkoxyalkyl, alkylthio, thioallcyl groups carboxylic acid
derivatives, or sulfonic acid derivatives; the linking group represented by -
(CII2)n-O- may be attached either through nitrogen atom or through carbon
atom where n is an integer ranging from 1 - 4; Ar rep'reserits an optionally
substituted divalent single or fused aromatic or heterocyclic group; R4
represents hydrogen atom, hydroxy, alkoxy, halogen, lower alkyl, optionally
substituted aralkyl group or forms a bond together with the adjacent group R5;
RS represents hydrogen, hydroxy, alkoxy, halogen, lower alkyl group, acyl,
optionally substituted aralkyl or RS forms a bond together with R4; R6 may be
hydrogen, optionally substituted groups selected from alkyl, cycloalkyl, aryl,
aralkyl, alkoxyalkyl, alkoxycarbonyl, aryloxycarbonyl, alkylaminocarbonyl,
arylaminocarbonyl, acyl, heterocyclyl, heteroaryl, heteroaralkyl groups, with
a
provision that R6 does not represent hydrogen when R' represents hydrogen or
lower alkyl group; R' may be hydrogen or optionally substituted groups
selected from. alkyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl,
heteroaralkyl groups; Y represents oxygen or NRB, where R8 represents
2'0 hydrogen, allcyl, aryl, hydroxyallcyl, arallcyl, heterocyclyl, heteroaryl,
heteroaralkyl groups; R7 and R$ together may form a substituted or
unsubstituted 5 or 6 membered cyclic structure containing carbon atoms,
which may optionally contain one or more heteroatoms selected from oxygen,
sulfur or nitrogen. We have also described the processes for preparing the
compounds of formula (II).
The pharmaceutically acceptable salts of the general formula (I) have
significant formulation and bulk handling advantages in view of the their
stability.
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Objective of the Invention
The present invention provides pharmaceutically acceptable salts of (3-
aryl-a-oxysubstituted alkylcarboxylic acids of the formula (I), their
derivatives, their analogs, their tautomeric forms, their stereoisomers, their
polymorphs, their pharmaceutically acceptable solvates and pharmaceutical
compositions containing them or their mixtures having good stability and
solubility, which can be used for the treatment and ! or prophylaxis of
diseases
related to increased levels of lipids, especially to treat hyperlipidemia, and
for
the treatment of type II diabetes, impaired glucose intolerance, leptin
resistance, atherosclerosis, disorders related to Syndrome X such as
hypertension, obesity, insulin resistance, coronary artery disease and other
cardiovascular disorders, renal diseases including glomerulonephritis,
glomerulosclerosis, nephrotic syndrome, hypertensive nephrosclerosis,
nephropathy; retinopathy, disorders related to endothelial cell activation,
psoriasis, polycystic ovarian syndrome (PCOS), dementia, diabetic
complications, eating disorders, osteoporosis, inflammatory bowel diseases,
myotonic dystrophy, pancreatitis, retinopathy, arteriosclerosis; xanthoma or
cancer with better efficacy, potency and lower toxicity.
The present invention provides pharmaceutically acceptable salts of (3-
aryl-a-oxysubstituted alkylcarboxylic acids of the formula (I) and their
derivatives, their analogs, their tautomeric forms, their stereoisomers, their
o polymorphs, their pharmaceutically acceptable solvates and pharmaceutical
compositions containing them or their mixtures which may have agonist
activity against PPARoc and / or PPARy, and optionally inhibit HMG CoA
reductase, in addition to agonist activity against PPARa and / or PPAR~y.
The present invention provides pharmaceutically acceptable salts of (3-
aryl-a-oxysubstituted alkylcarboxylic acids of the formula (I) and their
derivatives, their analogs, their tautomeric forms, their stereoisomers, their
polymorphs, their pharmaceutically acceptable solvates and pharmaceutical
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compositions containing them or their mixtures having enhanced activities,
without toxic effect or with reduced toxic effect.
The present invention provides a process for the preparation of
pharmaceutically salts of (3-aryl-a-oxysubstituted alkylcarboxylic acids and
their derivatives of the formula (I) as defined above, their analogs, their
tautomeric forms, their stereoisomers, their polyrriorphs and their
pharmaceutically acceptable solvates.
. The present invention provides pharmaceutical compositions containing
compounds of the general formula (I), their analogs, their derivatives, their
tautomers, their stereoisomers, their polymorphs, solvates or their mixtures
in
combination with suitable carriers, solvents, diluents and other media
normally employed in preparing such compositions.
Detailed Description of the Invention
The present invention relates to pharmaceutically acceptable salts
having the general formula (I)
O
P
1' ~° cn
,4~0 \ I ~ ORS
their derivatives, their analogs, their tautomeric forms, their'
stereoisoniers,
their polymorphs, wherein Rl represents hydrogen, alkyl or. aryl group; M
represents. a counter ion or a moiety which forms a pharmaceutically
acceptable salt; p is an integer ranging from 1 to 2; A represents a cyclic
structure given below
RZ O O O
N Ni \ Ni Ni
3
N ~ ~ N~R4 R ~ ~ N~R4 R4 N~R3
R3
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wherein R~ and R3 may be same or different and independently represent
hydrogen, halogen, hydroxy, vitro, cyano, alkyl or alkoxy group; R4 represents
hydrogen, ~ halogen, hydroxy, vitro, cyano, azido, formyl or unsubstituted or
5 substituted groups selected from alkyl, cycloalkyl, alkoxy, aryl,
heterocyclyl,
heteroaryl, amino, monoalkylamino, diallcylamino or alkoXyalkyl groups. ,
Suitable groups represented by Rl may be selected from hydrogen,
linear or branched (Cl-C6)alkyl group, such as methyl, ethyl, ~n-propyl,
10 isopropyl, n-butyl, isobutyl, t-butyl, ~n-pentyl, isopentyl, hexyl and the
like;
aryl group such as phenyl or naphthyl.
Suitable groups represented by RZ and R3 may be selected from
hydrogen, halogen atom such as fluorine, chlorine, bromine 'or iodide;
hydroxy, vitro, cyano, linear or branched (CI-C6)alkyl group, such as methyl,
ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, isopentyl,
hexyl,
heptyl and the lilce; linear or branched (C~-C6)alkoxy group, such as methoxy,
ethoxy, n-propoxy, isopropoxy and the like.
Suitable groups represented by R4 may be selected from hydrogen,
halogen, hydroxy, vitro, cyano, azido= formyl or unsubstituted or substituted;
linear or branched (C~-C6)alkyl group, such as methyl, ethyl, n-propyl,
isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, hexyl and the
like;
unsubstituted or substituted, linear or branched (C1-C6)alkoxy group, such as
rnethoxy, ethoxy, n-propoxy, isopropoxy and the like; cyclo(C3-C6)alkyl group
such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl~ and the like, the
cycloalkyl group may be substituted; aryl group such as phenyl or naphthyl,
the aryl group may be substituted; heteroaryl group such as pyridyl, thienyl,
furyl, pyrrolyl, oxazolyl, thiazolyl, iinidazolyl, oxadiazolyl, tetrazolyl,
benzopyranyl, benzofuranyl and the like, the heteroaryl group may be
substituted; heterocyclyl groups such as aziridinyl, pyrrolidinyl,
morpholinyl,
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piperidinyl, piperazinyl and the like, the heterocyclyl group may be
substituted; amino; monoalkylamino group such ~as NHCH3, ,NHCzHs,
NHC3H7, NHC6H13, and the like, which may be substituted; diallcylamino
group such as N(CH3)a, NCH3(CaHs), N(C2H5)2 and the like, which may be
substituted; allcoxyalkyl group such as methoxymethyl, ethoxymethyl,
methoxyethyl, ethoxyethyl and the like, which may be substituted. The
substituents may be selected from halogen atom such as fluorine, chlorine,
bromide or iodine; alkyl group such as methyl, ethyl, isopropyl, n-propyl, n-
butyl and the like.
Suitable groups represented by M may be selected from sodium, Mg,
calcium, potassium, I,i, glucamine, N-methyl glucamine; N-octyl glucamine,
dicyclohexylamiile, t-butyl amine, methyl benzylamine,
tris(hydroxymethyl)ainino methane (tromethamine), phenyl glycinol, lysine,
arginine, metformin, aminoguanidine, aminoguanidine hydrogen carbonate,
imidazole,, piperazine, dimethyl piperazine, pyrrolidine, berizylamine, phenyl
glycine methyl ester, phenylalanine benzyl ester or morpholine.
Particularly useful compounds according to the present invention include
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid phenyl glycinol salt;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid phenyl glycinol salt;
(-) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid phenyl glycinol salt;
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid methyl benzylamine salt;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]=2-
ethoxypropanoic acid methyl benzylamine salt;
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i2
(-) . 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid methyl benzylamine salt;
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid dicyclohexylamine salt;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoXy]phenyl]-2-
ethoxypropanoic acid dicyclohexylamine salt;
(-) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid dicyclohexylamine salt;
(~) , 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid lysine salt;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazoliii-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid lysine salt;
(-) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid lysine salt;
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid tris (hydroxymethyl)amino methane salt ;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)'ethoxy]phenyl]-2-
ethoxypropanoic acid tris (hydroxymethyl)amino methane salt ;
(-) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanaic acid tris (hydroxymethyl)amino methane salt ;
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4=dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid N-octyl glucamine salt;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquiriazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid N-octyl glucamine salt;
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(-) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoXypropanoic acid N-octyl glucaxnine salt;
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid N-methyl glucamine salt;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid N-methyl glucamine salt;
(-) 3-[4-[2.-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid N-methyl glucamine salt;
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-~-
ethoxypropanoic acid amino guanidine hydrogen carbonate salt;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid amino guanidine hydrogen carbonate salt;
(-) 3-[4-[2=(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid amino guanidine hydrogen carbonate salt ;
(~) 3-[4-[2-(~-Ethyl-4-oxo-3,4-dihydroquiiiazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid lithium salt;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid lithium salt;
3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid lithium salt;
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid arginine salt;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid arginine salt ;
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(-) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazoliri-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid arginine salt ;
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid metformin salt;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid metformin salt ;
(-) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid metfornZin salt ;
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid imidazole salt;
(+) ~ 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid imidazole salt;
(-) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid imidazole salt;
(~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquilzazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid magnesium salt ;
(+) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]=2-
ethoxypropanoic acid magnesium salt ;
(-) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoXypropanoic acid magnesium salt.;
(~) 3-[4-[2-(2-Morpholinyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-
2-ethoxypropanoic acid magnesium salt ; .
(+) 3-[4-[2-(2-Morpholinyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-
2-ethoxypropanoic acid magnesium salt;
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(-) 3-[4-[2-(2-Morpholinyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-
2-ethoxypropanoic acid magnesium salt;
(~) 3-[4-[2-(2-Piperidinyl-4-oxo-3,4-dihyd~oquinazolin-3-yl)ethoxy]phenyl]-
5 2-ethoxypropanoic acid magnesium salt ;
(+) 3-[4-[2-(2-Piperidinyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-
2-ethoxypropanoic acid magnesium salt ;
(-) 3-[4-[2-(2-Piperidinyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid magnesium salt ;
(~) 3-[4-[2-(2-Azido-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid magnesium salt ;
(+) , 3-[4-[2-(2-Azido-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid magnesium salt;
(-) 3-[4-[2-(2-Azido-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid magnesium salt ;
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid magnesium salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid magnesium salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid magnesium salt ;
(~) 3-[4-[2-(1,5-I~imethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid magnesium salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid magnesium salt ;
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(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[~;3-
d]pyrimidin-6-yl)eth'oxy]phenyl]-2-ethoxypropionic acid rizagnesium salt ;
(~) ~ 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid potassium salt ;
(+) 3-[4-[2-(1-Methyl=5-ethyl-7-oxo-3-propyl-6~7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid potassium salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid potassium salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4;3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid potassium salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid potassium salt ;
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3=
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid potassium salt ;
(~) 3-[4-[2-(1-Methyl-5=ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid calcium salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid calcium salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid calcium salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropi~onic acid calcium salt ;
(+) 3-[4=[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid calcium salt ;
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(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid calcium salt;
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lithium salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyriinidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lithium salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lithium salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrilnidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lithium salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lithium salt ;
(-) ' 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidiri-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lithium salt ;
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-lgi-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid sodium salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4;3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid sodium salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4;3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid sodium salt ;
(~) ' 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidiii-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid sodium salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid sodium salt ;
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(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1~I-pyrazolo[4,3-
d]pyririzidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid sodium salt ;
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)athoxy]phenyl]-2-ethoxypropionic acid arginine salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oXO-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid arginine salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid arginine salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethbxy]phenyl]-2-ethoxypropionic acid arginine salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid arginine salt ;
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid arginine salt ;
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidiiz-6-yI)ethoxyjphenyl]-2-ethoxypropionic acid rriethyl benzylamine
salt;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-lei-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid methyl benzylamine
salt;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid rilethyl benzylamiine
salt;
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(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid methyl benzylamine
salt;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid methyl benzylamine
salt; ,
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro=1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid W ethyl benzylamine
salt;
(~) 3-[4-[~-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid S-(+)-phenylglyciuol
salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid S-(+)-phenylglycinol
salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid S-(+)-phenylglycinol
salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid S-(+)-phenylglycinol
salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid S-(+)-phenylglycinol
salt ;
3-[4-[2-(1,5-Dimethyl-7-ono-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid S-(+)-phenylglycinol
salt ;
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(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidiii-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid amirioguanidine salt
5 (+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7=dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminoguanidine salt
(-) 3-[4-[2=(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)eth'oxy]phenyl]-2-ethoxypropionic acid aW inoguanidine salt
10 ;
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminoguanidine
hydrogen carbonate salt ;
15 (+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminoguanidine
hydrogen carbonate salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminoguanidiile
20 hydrogen carbonate salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,'7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminoguanidine
hydrogen carbonate salt ; ~ .
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminoguanidine
hydrogen carbonate salt ;
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(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminoguanidiile
hydrogen carbonate salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4;3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aininoguanidine salt
(+) ' 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminoguanidine salt
;
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminoguanidine salt
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4;3-
d]pyrirriidill-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid tromethamine salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid tioriiethamW a salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid tromethamine salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid tromethamine salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid tromethamine salt ;
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro=1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid tromethamine salt ;
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(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid dicyclohexylamine
salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid dicyclohexylamine
salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypiopionic acid dicyclohexylamine
salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid dicyclohexylam~ne
salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid dicyclohexylamine
salt ;
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid dicyclohexylamine
salt ; ~ .
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-octylglucamine
salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-octylglucamine
salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyriniidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-octylglucamine
salt ; ~ ~.
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(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7=dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-octylglucamine
salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-octylglucamine
salt ;
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-octylglucamine
salt ;
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[~,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N=methylglucamine
salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-inethylglucamine
salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-methylglucamine
salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-methylglucamine
salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-niethylglucamine
salt ;
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(-) 3-[4-[2-(1,5-Dimethyl-7-oa~o-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl)-2-ethoxypropionic acid N-methylglucamine
salt ; '
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyriinidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid metforrilin salt
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4;3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid metformin salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)'ethoxy]phenyl]-2-ethoxypropionic acid metformin salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4.,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid metformin salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid metformin salt ;
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoXy]phenyl]-2-ethoxypropionic acid metformin salt ;
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lysine salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1~I-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lysine salt ; ,i_
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxyp'ropionic acid lysine salt ;
(~) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoXy]phenyl]-2-ethoxypropionic acid lysine salt ; .
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyriinidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lysine salt ;
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(-) 3-[4-[2-(1,5-Dimethyl-7-ox~o-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lysine salt ;
(~) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
5 d]pyrimidin-6-yl)ethoxy]phenyl]-B-ethoxypropionic acid t-butylamine salt ;
(+) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid t-butylamine salt ;
(-) 3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4;3-
d]pyrimidin-6=yl)ethoxy]phenyl]-2-ethoxypropionic acid t-butylamine salt ;
(~) ~ 3-[4-[2-(1,5-Dimethyl-7-ox~o-3-propyl-6,7-dihydro-lei-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid t-butylamine salt ;
(+) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid t-butylarnine salt ;
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid t-butylamirie salt ;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid potassium salt ;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid potassium salt ;
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid potassium salt ;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid magnesium salt ;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid magnesium salt ;
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26
(-) . 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid magnesium salt ;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid phenyl glycinol salt;
(+) ~ 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid phenyl glycinol salt;
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid phenyl glycinol salt;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid sodium salt ;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid sodium salt ;
(-) ' 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid sodium salt ;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanQic acid t-butylamine salt ;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrirnidin-1-
yl]ethoxy]phenyl]propanoic acid t-butylamine salt ;
(-) . 2-Ethoxy-3-[4-[2-[2-ethyl=6-oxo-4-phenyl-1,6=dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid t-butylamine salt ;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid N-methyl glucamine salt;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid N-methyl glucamine salt;
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27
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid N-rizethyl glucamine salt;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid L-lysine salt ;
(+) 2-EthoXy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid L-lysine salt ;
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid L-lysine salt ;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid N-octyl glucainine salt;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid N-octyl glucamine salt;
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-diliydropyrimidin-1-
yl]ethoXy]phenyl]propanoic acid N-octyl glucamine salt;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid tris (hydroxymethyl)amino methane salt ;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1
yl]ethoxy]phenyl]propanoic acid tris (hydroxymethyl)amino methane salt ;
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]etho~y]phenyl]propanoic acid tris (hydroxymethyl)amino methane salt ;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid lithium salt ;
(+) .2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid lithium salt ;
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28
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid lithium salt ;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl=6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid calcium salt ;
(+) 2-Ethbxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid calcium salt ;
(-) ~ 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propaiioic acid calcium salt ;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyririzidin-1-
yl]ethoxy]phenyl]propanoic acid L-arginine salt ;
(+) . 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid L-arginine salt ;
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid L-arginine salt ;
(~) ~ 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid metformin salt ;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-.1-
yl]ethoxy]phenyl]propanoic acid metformin salt ;
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid metformin salt ;
(~) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid dicyclohexylamine salt ;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid dicyclohexylamine salt ;
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(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid dicyclohexylamine salt ;
(~) ' 2-Ethoxy-3-[4-[2-[2-ethyh6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid aminoguanidine salt ;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid aminoguanidine salt ;
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid aniinoguanidine salt ;
(~) ~ 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid methyl benzylamine salt;
(+) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid methyl benzylamine salt;
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-c~ihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid methyl benzylamine salt;
According to another feature of the present invention, there is provided
a process ~ for the preparation of pharmaceutically acceptable salts of the
formula (I) which comprises, reacting compound of the formula .(III'
O
(III)
~O
where all symbols are as defined earlier with a stoichiometric amount of an
appropriate base in the presence of a solvent at a temperature in the range of
10 °C to the boiling point of the solvent employed fox a period in the
range of
10 minutes to 30 hours.
The compound of the formula (III) used may be either optically pure
form or a racemic form. The base employed in the reaction may be selected
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from sodium hydroxide, sodium methoxide, potassium hydroxide, calcium
hydroxide, lithium hydroxide, magnesium hydroXide, glucamine, N-
methylglucamine, N-octylglucamine, dicyclohexylaniine, t-butylamine,
methyl benzylamine, tris(hydroxymethyl)aminomethane, phenyl glycinol,
5 lysine, arginine, metformin, aminoguanidine, aminoguanidine hydrogen
carbonate, imidazole, piperaziiie, dimethyl piperazirie, pyrrolidine,
benzylamine, phenyl glycine methyl ester, phenylalanine benzyl ester or
morpholine. The solvent employed may be selected from alcohols such as
ethanol, methanol, isopropanol, butanol and the like; lcetones such as
acetone,
10 diethyl ketone, methyl ethyl ketone or their mixtures; ethers such as
diethyl
ether, ether, tetrahydrofuran, dioxane, dibutyl ether and the like or DMF,
DMSO, xylene, toluene, ethyl acetate and the like or mixture thereof.
The pharmaceutically acceptable salts of the general formula (I) have
significant formulation and bulk handling advantages in view of the their
15 physicochemical properties and their stability.
Various polymorphs of a compound of general formula (I) forming part
of this invention may be prepared by crystallization of compound of formula
(I) under different conditions. For example, using different solvents commonly
20 used or their mixtures for recrystallization; crystallizatioris at
different
temperatures; various modes of cooling, ranging from very fast to very slow
cooling during crystallizations. Polymorphs may also be obtained by heating
or melting the compound followed by gradual or fast cooling. The presence of
polymorph's may be determined by solid probe NMR spectroscopy, IR
25 spectroscopy, differeritial scanning calorimetry, powder X-ray diffraction
or
such other techniques.
The stereoisomers of the compounds forming part of this invention may
be prepared by using compound of formula (1) in its single enantiomeric form
I in the process by resolving the mixture of stereoisomers by conventional
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31
methods. Some of the preferred methods include use of microbial resolution,
resolving the diastereomeric salts formed with optically pure bases such as
brucine, cinchona alkaloids and their derivatives, optically pure 2-alkyl
phenethyl amine, phenyl glycinol and the like. The diastereomeric salts may
be obtained in pure form by fractional crystallization. Commonly used
methods are compiled by Jaques et al in "Enantiorriers, Racemates and
Resolution" (Wiley Interscience, 1981).
Pharmaceutically acceptable solvates of the compounds of formula (I)
forming part of this invention may be prepared by conventional methods such
as dissolving the compounds of formula (I) in solvents such as water,
methanol, ethanol and the like, preferably water and recrystallizing by using
different crystallization techniques.
The present invention provides a pharmaceutical composition,
containing the compounds of the general formula (I) as defined above, their
derivatives, their analogs, their tautomeric forms, their stereoisomers, their
polymorphs, their pharmaceutically acceptable solvates in combination with
the usual pharmaceutically employed carriers, diluents and the like, useful
for
the treatment and / or prophylaxis of diseases such as hypertension, coronary
heart disease, atherosclerosis, stroke, peripheral vascular diseases and
related
disorders. These compounds are useful for the treatment of familial
hypercholesterolemia, hypertriglyceridemia, lowering . of atherogenic
lipoproteins, VLDL and LDL. The compounds of the present invention can be
used for the treatment of certain renal diseases including
glomerulonephri'tis,
glomerulosclerosis, nephrotic syndrome, hypertensive nephrosclerosis,
nephropathy. The compounds of general formula (I) are also useful for the
treatrizent/prophylaxis of insulin resistance (type II diabetes), leptin
resistance,
impaired glucose tolerance, dyslipidemia, disorders related to syndrome X
such as hypertension, obesity, insulin resistance, coronary heart disease, and
other cardiovascular disorders. These compounds may also' be useful as aldose
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32
reductase inhibitors, for improving cognitive functions in dementia, as
inflammatory agents, treating diabetic complications, disorders related to
endothelial cell activation, psoriasis, polycystic ovarian syndrome (PCO.S),
inflammatory bowel diseases, osteoporosis, myotonic dystrophy, pancreatitis,
retinopathy, arteriosclerosis, xanthoma and for the treatment of cancer. The
compounds of the present invention are useful in the treatment and/or
prophylaxis of the above said diseases in combination/concomittant with one
or more HMG CoA reductase inhibitors, hypolipidemic/ hypolipoproteinemic
agents such as fabric acid derivatives, nicotinic acid, cholestyramine,
colestipol, probucol or their combination. The compounds of the present
invention ' in combination with HMG CoA reductase inhibitors,
hypolipidemic/hypolipoproteinemic agents can be administered together or
within such a period to act synergistically. The IzTi~IG CoA reductase
inhibitors
nlay be selected from those used for the treatment or prevention of
hyperlipidemia such as lovastatin, provastatin, simvastatin, fluvastatin,
atorvastatin, cerivastatin and their analogs thereof. Suitable fabric acid
derivative may be gemfibrozil, ~ clofibrate, fenofibrate, ciprofibrate,
benzafibrate and their analogs thereof.
The present invention also provides a pharmaceutical composition,
containing the compounds of the general formula (I) as defined above, their
derivatives, their analogs, their tautomeric forms, their stereoisomers, their
polymorphs, their pharmaceutically acceptable solvates and one or more HMG
CoA reductase inhibitors, hypolipidemic a hypolipoproteineinic agents such.as
fabric acid derivatives, nicotinic acid, cholestyramine, colestipol, probucol
~ in
combination with the usual pharmaceutically employed carriers, diluents and
the like.
The pharmaceutical composition may be in the forms normally
employed, such as tablets, capsules, powders, syrups, solutions, suspensions
and the like, may contain flavorants, sweeteners etc. in suitable solid or
liquid
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33
carriers or. diluents, or in suitable sterile media to form injectable
solutions or
suspensions. Such compositions typically contain from 1 to 20 %, preferably
1 to 10 % by weight of active compound, the remainder of the composition
being pharmaceutically acceptable carriers, diluents or solvents.
Suitable pharmaceutically acceptable carriers include solid fillers or
diluents and sterile aqueous or organic solutions. The active ingredient will
be
present in such pharmaceutical compositions in the amounts sufficient to
provide the desired dosage in the range as described above. Thus, for oral
administration, the active ingredient can be combined with a suitable solid or
liquid carrier or diluent to form capsules, tablets, powders, syrups,
solutions,
suspensions and the like. The pharmaceutical compositions, may, if desired,
contain additional components such as flavourants, sweeteners, excipients and
the like. For parenteral administration, the active ingredient can be combined
with sterile aqueous or organic media to form injectable solutions ~or
1 S suspensions. For example, solutions in sesame or peanut oil, aqueous
propylene glycol and the like can be used, as well as aqueous solutions of
water-soluble pharmaceutically-acceptable acid addition salts or salts with
base of the compounds. Aqueous solutions with the active ingredient dissolved
in p~olyhydroxylated castor oil may also be used for injectable solutions. The
injectable solutions prepared in this manner can then be administered
intravenously, intraperitoneally, subcutaneously, or intramuscularly, with
intramuscular administration being preferred in humans.
For nasal administration, the preparation may contain the active
ingredient of the present invention dissolved or suspended in a liquid
carrier,
in particular an aqueous carrier, for aerosol application. The carrier may
contain additives such as solubilizing agents, such as propylene glycol,
surfactants, absorption enhancers such as lecithin (phosphatidylcholine) or
cyclodextrin or preservatives such as parabenes.
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34
Tablets, dragees or capsules having talc and / or a carbohydrate carried
binder arid the like are particularly suitable for any oral application.
Preferably, carriers for tablets, dragees or capsules include lactose, corn
starch
and / or potato starch. A syrup or elixir can be used in cases where a
sweetened vehicle can be employed.
A typical tablet production method is exemplified below °.
')<'alilet Production Example
a) 1) Active ingredient 30 g
2) Lactose 95 g
3) Corn starch 30 g
4) Carboxymethyl cellulose 44 g
5) MagnesiLUn stearate 1 g
200 g for 1000 tablets
The ingredients 1 to 3 are uniformly blended with water and granulated after
drying under reduced pressure. The ingredient 4 and 5 are' mixed well with the
granules and compressed by a tabletting machine to prepare 100'0 tablets each
containing 30 mg of active ingredient:
b) 1) Active ingredient30
g
2) Calcium phosphate 90
g
3) Lactose 40
g
4) Corn starch 35
g
5) Polyvinyl pyrrolidone 3.5
g
6) Magnesium stearate 1.5
g
200 g for 1000 tablets
The ingredients 1-4 are uniformly moistened with an aqueous solution
of 5 and granulated after cliying under reduced pressure. Ingredient 6 is
added
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and granules are compressed by a tabletting machine to prepare 1000 tablets
containing 30 mg of ingredient 1.
The compound of the formula (I) as defined above are clinically
5 administered to mammals, including man, via either oral, nasal, pulmonary,
transdermal or parenteral, rectal, depot, subcutaneous, intravenous,
intrauxethral, intramuscular, intranasal, ophthalmic solution or an ointment.
Administration by the oral route is preferred, being more convenient and
avoiding the possible pain and irritation of injection. However, ili
10 circumstances where the patient cannot swallow the medication, or
absorption
following oral administration is impaired, as by disease or other abnormality,
it is essential that the drug be administered parenterally. By eider route,
the
dosage is in the range of about 0.01 to about 100 ing / kg body weight of the
subject per day or preferably about O.Ol to about 30 mg / kg body weight per
15 day administered singly or as a divided dose. However, the optimum dosage
for the individual subject being treated will be determined by the person
responsible for treatment, generally smaller doses being administered
initially
and thereafter increments made to determine the most suitable dosage.
20 The invention is explained in detail in the examples given below
which are provided by way of illustration only and therefore should not be
construed to limit the scope of the invention.
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
25 , ethoxypropanoic acid was prepared according to the procedure given in WO
9910g501 as given below
A solution of [(2S)-N(1S)]-2-ethoxy-3-[4-[2-(2,-Ethyl-4-oxo-3,4-
dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxypropanoic acid-N-(2-
g
3) Lactose
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36
hydroxy-1-phenylethyl)propanamide (267 mg, 0.504 mmol) in a mixture of
11VT sulphuric acid and dioxane ! water was heated at 100 °C for 16 h.
The
reaction mixture was cooled to ca 25 °C and dioxane was removed under
reduced pressure. The remaining solution was cooled in an ice bath and the
white solid precipitated was filtered and dried to afford (-)-3-[4-[2-(2-ethyl-
4-
oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxypropanoic acid (170
mg, 82 %).
However, any other procedure for preparing (-)-3-[4-[2-(2-Ethyl-4-oxo
3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxypropanoic acid can be
used. (~) 3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]
2-ethoxypropanoic acid , and (+)-3-[4-[2-(2-Ethyl-4-oxo-3,4-
dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxypropanoic acid can be
prepared by a similar procedure described above.
Example-1
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2,-
ethoxypropanoic acid phenyl glycinol salt
O _
Ph
. I w NCO ~ /. ~ ~ . ~ OH
C-O NH
Et0
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid (2 g) and isopropanol (50 ml) was added to 250 ml four
necked round bottom flask fitted with a mechanical stirrer and reflux
condenser: The reaction mixture was slowly heated to 45-55 °C for
complete
dissolution of the. reaction mass. Phenyl glycinol (0.667 g) dissolved in
isopropanol (10 ml) was added to the reaction mixture at 45-55 °C in
about 10
min. under stirring. The progress of the reaction was maintained by gentle
reflux of reaction mixture at 75-85 °C for 10 h. The reaction mixture
was
cooled to room temperature and stirred for 5 h at room temperature. The
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37
precipitated product was filtered, dried at 60 °C for 2-3 h to afford
pure phenyl
glycinol salt of (-)-3-[4-[2-(2-ethyl-4-oxo-3,4-dihydroquinazolin-3-
yl)ethoxy]phenyl]-2-ethoxypropanoic acid as free flowing white crystalline
solid, (weighs about 2 g, yield 75 %, mp : 105 -107 °C, purity 99 % by
HPLC).
IR (KBr) ciri 1 : 3400-3300 (O-H stretch), 2922 (-C-H aliphatic stretch), 1670
(-COO- stretch), 1590(-CONH stretch), 1420 (-COO stretch).
1H NMR (200 MHz, DMSO-d6) 8 : 8.13 (d, J=7.89Hz, 1H), 7.82 (t, J=7.01Hz,
1H), 7.64 (d, J=8.21Hz, 1H), 7.50 (t, J= 7.26Hz, 2H), 7.4 (s, SH), 7.13 (d, J=
8.SOHz, 2H), 6.84 (d, J=B.SOHz, 2H),'4.47 (t, J=5.19Hz, 2H), 4.4 (d, 2H), 4.26
(t, J=5,19Hz, 2H), 3.99-3.84 (m, 1H), 3.60-3.40 (m, 1H), 3.40-3.20 (m, 2H),
3.06 (q, J= 6.96Hz, 2H), 2.88 (q, J= 6.64Hz, 2H), 1.32.(t, J= 7.17Hz, 3H),
1.02
(t, J= 6.96Hz, 3H).
Mass m/z .: 411 (M+ + 1), 137 (C$H11N0). Anal. Calcd : C31H37N3O6; % C
68.00; % H 6.76; % N 7.67, Found % C 67.80; % H 6.56; % N 7.57.
Example-2
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid R-(+) methyl benzylamine salt
.
O NH3
O
~ HBO ~ ~ , C_O~ , I W CH3
Et~
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoXy]phenyl]-2-
ethoxypropanoic acid (2.0 g) and isopropanol (50 m1) was added to 250 ml
four necked round bottom flask, fitted with a mechanical stirrer and reflux
condenser. The reaction mixture was slowly heated to 45-55 °C for
complete
dissolution of the reaction mass. R-(+) methyl benzylamine (0.589 g) in
isopropanol (10 m1) was added to the reaction mixture at 45-55 °C in
about 10
min. under stirring. The progress of the reaction was maintained by gentle
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38
reflux of reaction mixture at 75-85 °C for 10 h. The reaction mixture
was
4
cooled to room temperature and stirred for 12 h at room temperature. The
reaction mixture was cooled to -5 °C and maintained at that temperature
for 2
h under stirring. The precipitated product was filtered, dried at 60 °C
for 2 h to
afford pure (R)-(+)-methyl benzylamine salt of (-)-3-[4-[2-(2-ethyl-4-oxo-3,4-
dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxypropanoic acid as free
flowing off white amorphous solid, (weighs about 2 g, yield 80 %, mp : 137-
139 °C, purity 99 % by HPLC).
IR (KBr) crri 1 : 3400-3300 (N-H stretch), 3120 (-C-H aromatic), 2930 (-C-H
aliphatic), 1660 (-COO stretch), 1583 ,(-CONH stretch), 1400 (=COO stretch).
1H NMR (200 MHz; DMSO-d6) b : 8.13 (d; J=7.89Hz, 1H), 7.82. (t; J=7.OlHz,
1H), 7.64 (d, J=8.21Hz, 1H), 7.6-7.2 (m, SH), 7.50 (t, ~= 7.26Hz, 2H), 7.13
(d,
J= B.SOHz, 2H), 6.84 (d, J=8.SOHz, 2H), 4.47 (t, J=5.19Hz, 2H), 4.26 (t,
J=5.19Hz, 2H), 3.99-3.84 (m, 1H), 3.60-3.40 (m, 1H); 3.40-3:20 (m, 1H), 3.06
(q, J= 6.96Hz, 2H), 2.88 (q, J= 6.64Hz, 2H), 1.32 (t, J= 7.17Hz, 3H), 1.02 (t,
J= 6.96Hz, 3H), 1.0 (d, 3H).
Mass mlz : 411 (M+ + 1), 121 (C8H11N). Anal. Calcd. C31H37N3O5, %. C
70.05, % H 6.96; % N 7.90%; Found % C 69.82; % H 6.80; % N 7.70.
Example-3
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid dicyclohexylamine salt .
0
H
. <
Et0
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4~dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid (2 g) and isopropanol (50 ml) was added to 250 ml four
necked round bottom flask, fitted with a mechanical stirrer and reflux
condenser. The reaction mixture was slowly heated to 45-55 °C for
complete
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39
dissolution of the mass. Dicyclohexylamine (0.88 g) dissolved in isopropanol
(10 ml) was added to the reaction mixture at 45-55 °C in about 10 Thin.
under
stirring. The progress of the reaction was maintained by gentle reflux of
reaction mixture at 75-85 °C for 10 h. The reaction mixture was cooled
to 0-5
°C and maintained for 2 h under stirring. The precipitated product was
filtered,
dried at 60 °C fox 2-3 h to afford pure dicyclohexylamine salt of (-)-3-
[4-[2-(2-
ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxypropanoic
acid as free flowing off white crystalline solid, (weighs about 2.2 g, yield
76
% mp : 152-153 °C; purity 99 % by HPLC).
IR (I~Br) cm 1 : 3400-3300 (-N-H stretch), 3100 (C-H, aromatic), 2930 (=C-H,
aliphatic), 1670 (-COO stretch), 1597 °(-CONH stretch), 1400 (-COO
stretch).
1H NMR (200 MHz, DM80-d6) b : 8.13 (d, 3=7.89Hz, 1H), 7.82 (t, J=7.OlHz,
1H), 7.64 (d, J=8.21Hz, 1H), 7.50 (t, J= 7.26Hz, 2H), 7.13 (d, J= 8.SOHz, 2H),
6.84 (d, J=8.SOHz, 2H), 4.47 (t, J=5.19Hz, 2H), 4.26 (t, J=5.19Hz, 2H), 3.99-
3.84 (m, 1H), 3.60-3.40 (m, 1H), 3.40-3.20 (m, 1H), 3.06 (q, J= 6.96Hz, 2H),
2.88 (q, J= 6.64Hz, 2H), 2.6-0.8 (m, 22H), 1.32 (t, J= 7.17Hz, 3H), 1.02 (t,
J=
6.96Hz~ 3H).
Mass mlz : 411 (M+ + 1), 181 (Cl2HasN). Anal. Calcd :°C35H49N3O5;
% C
71.06;%H8.29;%N7.10;Found°/~C70.89;%H8.10;%N6.95.
Example 4 .
(-)-3-[4-[Z-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl-2-
ethoxypropanoic acid lysine salt
O
O NHs
NCO \ / O~0 . (CH2)4
H-,-NH2
Et0
COOH
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid (2 g) and isopropanol (50 ml) was added to 250 ml four
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necked round bottom flask, fitted with a mechanical stirrer and reflux
condenser. The reaction mixture was slowly heated to 45-55 °C for
complete
dissolution of the mass. Lysine (0.8 g) dissolved in isopropanol (10 ml) was
added to the reaction mixture at 45-55 °C in about 10 min. under
stirring. The
5 progress of the reaction was maintained by the gentle reflux of reaction
mixture at 75-85 °C for 10 h. The reaction mixture was cooled to room
temperature and stirred for 12 h at room temperature. The precipitated product
was filtered, dried at 60 °C for 2-3 h to afford pure lysine salt of (-
)-3-[4-[2-(2-
ethyl-4-oxo-3,4-dihydroquinazolin=3-yl)ethoxy]phenyl]-2-ethoxypropanoic
10 acid as free flowing off white amorphous solid (weights about 2 g, yield :
75
%, mp : 152-155 °C purity 99 % by HPLC).
IlZ (KBr) cm 1 : 3400-3300 (N-H stretch), 3120 (-C-H, aromatic), 2930 (-C-H
aliphatic), 1670 (-COO stretch), 1584 (-CONH stretch), 1407 (-COO stretch).
1H NMR (200 MHz, DMSO-d6) 8 : 8.13 (d, J=7.89Hz, 1H), 7.82 (t, J=7.01Hz,
15 1H), 7.64 (d, J=8.21Hz, 1H), 7.50 (t, J= 7.26Hz, 2H), 7.13 (d, J= 8.50Hz,
2H),
6.84 (d, J=8.50Hz, 2H), 4.47 (t, J=5.19Hz, 2H), 4.26 (t, J=5.19Hz, 2H), 3.99-
3.84 (m, 1H), 3.60-3.40 (m, 1H), 3.40-3.20 (m, 1H), 3.19 (t, 2H), 3.06 (q, J=
6.96Hz, 2H), 2.88 (q, J= 6.64Hz, 2H), 2.63 (t, 2H), 1.52 (m, 2H), 1.32 (t, J=
7.17Hz, 3H), 1.32 (m, 2H), 1.02 (t, J= 6.96Hz, 3H).
20 Mass m!z : 411 (M+ + 1), 146 (C6H14NZO2). Anal. Calcd : C~9H4pN4O7 ; % C
62.5; % H 7.19; % N 10.07; Found % 062.35; % H 7.10; % N 9.89.
Example 5
(-)-3-[4-[2-(2-Ethyl=4-oxo-3,4=dihydroquinazolin-3-yl)ethoxy] phenyl]-2-
25 ethoxypropanoic acid tris (hyd~oxymethyl)amino methane salt
O
HO OH
NCO ~ / O O . ~
C-O 0.,. Hs~OH
N~ Et0
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41
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoXypropanoic acid (2 g) and isopropanol (50 rizl) was added to 250 ml four
necked round bottom flask, fitted with a mechanical stirrer and xeflux
condenser. The reaction mixture was slowly heated to 45-55 °C for
complete
dissolution of the mass. Tris (hydroxymethyl) amino methane (0.59 g)
dissolved in isopropanol (10 ml) was added to the reaction mixture at 45-55
°C in about 10 min. under stirring. The progress of the reaction was
maintained by gentle reflux of reaction mixture at 75-85 °C for 10 h.
The
reaction mixture was cooled to room temperature and stirred for 12 h. Distill
off the isopropanol on rotavapor bath at 45-55 °C under vacuum. The
product
could i~ot be isolated as a fine solid, because of hygroscopic nature and
obtained as a sticky gummy mass, (weighs about 2 g, yield 77 %, purity 99
by HPLC).
IR (I~Br) cm ~ : 3400-3300 (N-H stretch), 3100 (C-H aromatic), 2937 (-C-H
aliphatic), 1670 (-COO stretch), 1584 (-CONH stretch), 1395 (-COO stretch).
1H NMR (200 MHz, DMSO-d6) 8 : 8.13 (d, J=7.89Hz, 1H), 7.82 (t, J=7.01Hz,
1H), 7.64 (d, J=8.21Hz, 1H), 7.50 (t, J= 7.26I3z, 2H), 7.13 (d, J= 8.SOHz,
2H),
6.84 (d, J=8.SOHz, 2H), 4.47 (t, J=5.19Hz, 2H), 4.26 (t, J=5.19Hz, 2H), 3.99-
3.84 (m, 1H), 3.8 (s, 6H), 3.60-3.40 (m, 1H), 3.40-3.20 (iii, 1H), 3.06 (q, J=
6.96Hz, 2H), 2.88 (q, J= 6.64Hz, 2H), 1.32 (t, J= 7.17Hz, 3H), 1.02 (t, J=
6.96Hz, 3H).
Mass. mlz : 411 (M+ + 1), 121 (C4H11N03).
Example 6
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid N-octyl glucamine salt
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42
CH2-NH2-C$H~~
O H OH
W NCO ~ ~ O_O . WO H
Et0 H OH
N H' OH
CH20H
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid (2 g) and isopropanol (50 ml) was added to 250 ml four
necked round bottom flask, fitted with a mechanical stirrer and reflux
condenser. The reaction mixture was slowly heated to 45-55 °C for
complete
dissolution of the mass. N-octyl glucarriine salt (1.42 g) dissolved ~in
isopropanol (10 ml) was added to the reaction mixture at 45-55 °C in
about 10
min. under stirring. The progress of the reaction was maintained by the gentle
reflux of reaction mixture at 75-85 , °C for 6 h. The reaction mixture
was
cooled to room temperature and stirred for 12 h. The precipitated product was
filtered and dried at 60 °C for 2-3 h to afford pure N-octyl glucamine
salt (-)-
3-[4-[2-(2-ethyl-4=oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxy
propanoic acid as free flowing off white crystalline solid, (weighs about 2.7
g,
yield 92 %, Trip : 114-116 °C, purity 99 % by HPLC).
IR (I~Br) cm 1 : 3400-3250 (-N-H stretch), 2926 (-C-H stretch), 2800-2200 (-
NH3 stretch), 1776 (-COO stretch), 1593 (-CONH stretch), 1410 (-COO
stretch).
1H NMR (200 MHz, DMSO-d6) 8 : 8.13 (d, J=7.89Hz; 1H), 7.82 (t, J=7.OlHz,
1H),'7.64 (d, J=8.21Hz, 1H), 7.50 (t, ~= 7.26Hz, 2H), 7.13 (d, J= 8.50Hz, 2H),
6.84 (d, J=8.50Hz, 2H), 4.47 (t, J=5.19Hz, 2H), 4.26 (t, J=5.19Hz, 2H), 4.2-
3.06 (m, 8H), 3.99-3.84 (m, 1H), 3.60-3.40 (m, 1H), 3.40-3.20 (m, 1H), 3.06
(q, J= 6.96Hz, 2H), 2.88 (q, J= 6.64Hz, 2H), 1.5 (d, 3H), 1.40-1.00 (m, 1~6H),
1.32 (t, J= 7.17Hz, 3H), 1.02 (t, J= 6.96Hz, 3H).
Mass m/z ~: 411 (M+ +1), 293 (C14H3iNOs). Anal. Calcd. C37,H57N3O1o, % C
63.15; % H 8.10; % N 5.97; Found %C 63.01; %H 7.91; %N 5.73.
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Example 7
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid N-methyl glucamine salt
O
CHZ-NH2-CH3
O H OH
\ NCO ~ ~ 0_0 . HO H
~ ~~ Et0 H OH
H OH
CH20H
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxyproparloic acid (2 g) and isopropanol (50 ml) was added to 250 ml four
necked round bottom flask, fitted with a mechanical stirrer and reflux
condenser. The reaction mixture was slowly heated to 45-55 °C for
complete
dissolution of the mass. N-methyl glucamine (0.95 g) in isopropanol (20 ml)
was added to the reaction mixture at 45-55 °C in about 10 min. under
stirring.
The progress of the reaction was maintained by gentle reflux of reaction
mixture at 75-85 °C for 5 h. The reaction mixture was cooled to RT and
stirred
for 12 h at room temperature. The precipitated product was filtered, and dried
at 60 °C for 2-3 h to afford pure N-methyl glucarriine salt of (-)-3-[4-
[2-(2-
ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxypropanoic
acid as free flowing off white crystalline solid, (weighs about 2.75 g, yield
: 93
%, mp : 114-116 °C, purity 99 % by HPLC).
IR (KBr) cm 1 : 3350-3300 (-NH, -OH stretching), 2960 (C-H stretch), 1670 (-
COO stretch), 1887 (-CONH stretch).'
1H NMR (200 MHz, DMSO-d6) 8 : 8.13 (d, J=7.89Hz, 1H), 7.82 (t, J=7.01Hz,
1H), 7.64 (d, J=8.21Hz, 1H), 7.50 (t, J= 7.26Hz, 2H), 7.13 (d, J= 8.SOHz, 2H),
6.84 (d, J=8.SOHz, 2H), 4.47 (t, J=5.19Hz, 2H), 4.26 (t, J=5.19Hz, 2H), 4.0-
3.26 (m, $H), 3.99-3.84 (m, 1H), 3.60-3.40 (m, 1H), 3.40-3.20 (m, 1H), 3.06
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(q, J= 6.96Hz, 2H), 2.88 (q, J= 6.64Hz, 2H), 2.4 (s, 3H), 1.32 (t, J= 7.17Hz,
3H), 1.02 (t, J= 6.96Hz~ 3H).
Mass m/z : 411 (M+ + 1), 195 (C7H17N0=). Anal. Cacld. C3oH43N3~10~ % C
59.5;%H7.10;%N6.94;Found%C59.25;%H6.9;%N6.74.
Example 8
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)etho~y]phenyl]-2-
ethoxypropanoic acid aminoguanidine hydrogen carbonate salt
0
O O ~ NIIH
\ ~ ~ ~ C~O~ . H3N.N~ 0. HCOO
Et0 H
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoXy]phenyl]-2-
ethoxypropanoic acid (2 g) and isopropanol (50 ml) was added to 250 ml four
necked round bottom flask, fitted with a mechanical stirrer and reflux
condenser. The reaction mixture was slowly heated to 45-55 °C for
complete
dissolution of the mass. Aminoguanidine hydrogen carbonate (0.662 g) ll1
isopropanol (20 ml) was added to the reaction mixture at 45-55 °C in
about 10
min. under stirring. The progress of the reaction was maintained by gentle
reflux of reaction mixture at 75-85 °C for 10 h. The reaction rilixture
was
cooled to room temperature and stirred for 12 h at room terilperature. Distill
off the isopropanol on rotavapor water bath at 45-55 °C under stirring.
The
product could not be isolated as fme solid, because of hyg~oscopic nature and
obtained as sticky gummy mass (weighs about 2 g, yield 75 %, purity 99 % by
HPLC).
IR (KBr) cm 1 : 3400-3300 (N-H~ stretch), 2970 (=C-H, aliphatic stretch), 1675
(-COO stretch), 1595 (-CONH stretch), 1392 (-COO stretch).
1H NMR (200 MHz, DMSO-d6) 8 : 8.13 (d, J=7.89Hz, 1H), 7.82 (t, J=7.Olliz,
1H), 7.64 (d, J=8.21Hz, 1H), 7.50 (t, J= 7.26Hz, 2H), 7.13 (d, J= 8.50Hz, 2H),
6.84 (d, J=8.50Hz, 2I3), 4.47 (t, J=5.19Hz, 2H), 4.26 (t, J=5.19Hz, 2H), 3.99-
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3.84 (m, 1H), 3.60-3.40 (m, 1H), 3.40-3.20 (m, lI~), 3.06 (q, J= 6.96Hz, 2H),
2.88 (q, J= 6.64Hz, 2H), 2.4 (s, 6H), 1.32 (t, J= 7.17Hz, 3H), 1.02 (t, J=
6.96Hz, 3H).
Mass m/z : 411 (M~ +1), 136 (CZH8N402).
5
Exaanple 9
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid lithium salt
o _
O ~ ~ O-po . Lid
Et0
10 (-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid (5 g) and isopropanol (100 ml) was added to 250 ml
four necked round bottom flask, fitted with a mechanical stirrer and reflux
condenser. The reaction mixture was, slowly heated to 45-55 °C for
complete
dissolution of the mass. Lithium hydroxide hydrate (0.51 g) dissolved in water
15 (20 ml) was added to the reaction mixture and heated to 75-85 °C for
12 h and
monitor the progress of the reaction. The reaction mixture was cooled to room
temperature and stirred for 12 h at room temperature. Distill off the
isopropanol on rotavapor water bath at 45-55 °C under vacuum.
Isopropanol
(50 ml) was added and the precipitated product was filtered, dried at 60
°C for
20 2-3 to afford pure lithium salt of (-)-3-[4-[2-(2-ethyl-4-oxo-3;4-
dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxypropanoic acid as off white
crystalline solid, (weighs about 4.5 g, yield : 90 %, mp, : 245-47 °C,
purity 99
by HPLC). .
IR (KBr) cm 1 : 3120 (-C-H aromatic), 2930 (-C-H aliphatic), 1660 (-COO
25 stretch), 1587 (-CONH stretch), 1400 (-COO stretch).
1H NMR (200 MHz, DMSO-d6) 8 : 8.13 (d, J=7.89Hz, 1H), 7.82 (t, J=7.OlHz,
1H), 7.64 (d, J=8:21Hz, 1H), 7.50 (t, J= 7.26Hz, 2H), 7.13 (d, J= 8.50Hz, 2H),
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6.84 (d, J=B.SOHz, 2H), 4.47 (t, J=5.19Hz, 2H), 4.26 (t; J=5.19Hz, 2H), 3.99-
3.84 (m, 1H), 3.60-3.40 (m, 1H), 3.40-3.20 (m, 1H), 3.06 (q, J= 6:96Hz, 2H),
2.8~ (q, J= 6.64Hz, 2H); 1.32 (t, J= 7.17Hz, 3H), 1.02 (t, J= 6.96Hz, 3H).
Mass m/z : 411 (M+ + 1). Anal. Calcd : C23H25N205Li; % C 66.34; % H 6.00;
% N .6.73; Found % C 66.12; % H 5.99; % N 6.53.
Example 10
(-)-3-[4-[2-(B-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid magnesium salt
O
~ N~o
Et0
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid (6 g) and isopropyl alcohol (60 ml) was added to 250 ml
four necked round bottom flask, fitted with a mechanical stirrer and reflux
condenser. The reaction mixture was slowly heated to 45-55 °C for
complete
dissolution of the mass. Magnesium hydroxide (0.423 g) was added to the
reaction mixture and heated to 75-85 °C for 12 h and monitored the
progress
of the reaction. The reaction mixture was cooled to RT and stirred for 12 h at
room temperature. Distill off the isopropanol on rotavapor water bath at 45-55
°C under vacuum and added isopropanol (5 ml) to the glassy residue and
stirred for 10 min. The precipitated product was filtered, dried at 60
°C for 2-3
h to afford pure magnesium salt of (-)-3-[4-[2-(2-ethyl-4-oxo-3,4-
dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxypropanoic acid as free
flowing off white amorphous solid, (weighs about 5 g, yield : 80 %, nip : >25U
°C, purity 99% by HPLC).
IR (KDr) cm 1 : 3120 (-C-H aromatic), 2930 (-C-H aliphatic), 1660 (-COO
stretch), 1587 (-CONH stretch), 1400 (-COO stretch).
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1H NMR (200 MHz, DMSO-d6) 8 : 8.13 (d, J=7.89Hz, 1H), 7.82 (t, J=7.OlHz,
1H), 7.64 (d, J=8.21Hz, 1H), 7.50 (t, J= 7.26Hz, 2H), 7.13 (d, J= 8.SOHz,
2~I),
6.84 (d, J=8.SOHz, ZH), 4.47 (t, J=5.19Hz, 2H), 4.26 (t, J=5.19Hz, 2H), 3.99
3.84 (m, 1H), 3.60-3.40 (m, 1H), 3.40 -3.20 (m, 1H), 3.06 (q, J= 6.96Hz, 2H),
2.88 (q, J= 6.64Hz, 2H), 1.32 (t, J= 7.17Hz, 3H), 1.02 (t, J= 6.96Hz, 3H).
Mass m/z : 411 (M+ + 1). Anal. Calcd : C46HSON4010Mg~ % C 65.63; % H
5.94; % N 6.65; Found % C 65.50; % H 5.75; % N 6.50.
Example 11
Form ~
(-)-3-[4-(B-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy] phenyl]-2-
ethoxypropanoic acid arginine salt
O - H NH2
W NCO ~ / IOCI-OO , HN~N~C02H
Et0 ~ NH3
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2
ethoxy propanoic acid (4.1 g), isopropanol (50 ml) was added to 250 ml four
necked round bottom flask, fitted with a mechanical stirrer and reflux
condenser. The reaction mixture was slowly heated to 45-55 °C in about
10
min. under stirring. L-Arginine (1.74 g) was dissolved in DM water (5 ml)
and added' at 45-55 °C under stirring. Maintained the gentle reflux of
reaction
mixture at 75-85 °C for 10 h and monitored the progress of the
reaction. The
reaction mixture was cooled to room temperature and stirred for 12 h at room
temperature. Distill off the isopropanol on rotavapor bath at 45-55 °C
under
. vacuum and added isopropanol (5 ml) to the glassy residue and stirred for 10
min. . The precipitated product was filtered, , dried at 60 °C for 2-3
h to afford
form I of pure arginine salt of ~ (-)-3-[4-[2-(2-ethyl=4-oxo-3,4
dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxy propanoic acid as off white
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amorphous solid (weights about 4.67 g, yield : 80%, W ~p : 215 °C,
purity 99
by HPLC).
IR (KBr) cm 1 : 34f0-3300 (N-H stretch), 3120 (-C-H, aromatic), 2930 (-C-H
aliphatic), 1670 (-COO stretch(, 1584 (-CONH stretch), 1407 (-COO stretch).
1H NMR (200 MHz, DMSO-d6) b : 8.13 (d, J=7.89Hz, 1H), 7.82 (t, J=7.OlHz,
1H), .7.64 (d, J=8.21Hz; 1H), 7.50 (t, J= 7.26Hz, 2H), 7.13 (d; J= 8.50Hz,
2H),
6.84 (d, J=8.SOHz, 2H), 4.47 (t, J=5.19Hz, 2H), 4.26 (t, J=5.19Iiz, 2H), 3.99-
3 . 84 (m, 1 H), 3. 8-3 .2 (m, 7H); 3 .60-3 .40 (m, 1 H), 3 .40-3 .20 (m, 1
H), 3.19 (t,
2H), 3.06 (q, J= 6.96Hz, 2H), 2.88 (q, J= 6.64Hz, 2H), 2.63 (t, 2H), 1.52 (m,
2H), 1.32 (t, J= 7.17Hz, 3H), 1.32 (m, 2H), 1.02 (t, J= 6.96Hz, 3H).
Mass m/z : 411 (M+ + 1). Anal. Calcd : C29H4oN607 ; .% C 59.57; % H 6.84;
N 14.38; Found % C 59.40; % H 6.00; % N 13.28.
Form a
(-)-2-Ethoxy-3-[4-[2-(2-ethyl-4-oxo-3,4-dihydro-3-quinazolinyl)ethoxy]
phenyl]propionic acid ( 140 g), isopropanol (2.8 L) was added to 5 L four
necked round bottom flask, fitted ' with a mechanical stirrer and reflux
condenser. The reaction mixture was slowly heated to 65-75 °C in about
10
min. under stirring. L-Arginine (59.2 g) dissolved in demineralized (DM)
water (296 ml) was added at 65-75 °C under stirring. After addition of
the
arginine solution, the reaction mixture becomes clear and precipitation
appeared immediately in the reaction mixture. Maintained the gentle reflux of
reaction mixture at 75-85 °C for 4-8 h and monitored the progress of
the
reaction. The reaction mixture was cooled to room temperature and stirred for
2 h at room temperature. The precipitated product was filtered, dried at 60-65
°C for 5-6 h, till moisture content (MC) reached <1 %, to afford form
II of
pure arginiiie salt of (-)-3-[4-[2-(2-ethyl-4-oxo-3,4-dihydroquinazolin-3-
yl)ethoxy]phenyl]-2-ethoxypropanoic acid as white crystalline solid, (weights
about 1.80 g; yield : 90 %, Trip 216-220 °C, purity 99 % by HPLC).
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IR (I~Br) cm 1 : 3400-3300 (N-H stretch), 312,0 (-C-H, aromatic), 2930 (-C-H
aliphatic), 1712 (-COOH), 1670 (-COO stretch, -CONH stretch), 1584 (-
CONH stretch), 1407 (-COO stretch).
1H NMR (200 MHz, DMSO-d6) 8 : 8.13 (d, J=7.89Hz, 1H), 7.82 (t, J=7.OlHz,
1H), 7.64 (d, J=8.21Hz, 1H), 7.50 (t, J= 7.26Hz, 2H), 7.13 (d, J= 8.50Hz, 2H),
6.84 (d, J=8.50Hz, 2H), 4.47 (t, J=5.19Hz, 2H), 4.26 (t, J=5.19Hz~ 2H), 3.99-
3.84 '(m, 1H), 3.8-3.2 (m, 7H), 3.60-3.40 (rim, 1H), 3.40-3.20 (m, 1H), 3.19
(t,
2H), 3.06 (q, J= 6.96Hz, 2H), 2.88 (q, J= 6.64Hz, 2H), 2.63 (t, 2H), 1.52 (m,
2H), 1.32 (t, J= 7.17Hz, 3H), 1.32 (m, 2H), 1.02 (t, J= 6:96Hz, 3H).
Mass mlz : 411 (M+ + 1). Anal. Calcd : Ca9H4oN607 ; % C 59.57; % H 6.84;
N 14.38; Found % C 59.40; % H 6.00; % N 13.28.
Example 12
(-)-3-[4-[2-(2-Azido-4-oXO-3,4-dihydroquinazoliri-3-yl)ethoxy]phenyl]-2- .
ethoxypropanoic acid anagnesium salt
0
W N~0 ~ / ~-OO M O+
9
N~N3 Et0
2
(-)-3-[4-[2-(2-Azo-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid (61 mg), dry methanol (5 ml) and magnesium hydroxide
(3.98 mg)~ was added to 50 ml one necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was refluxed at
80 °C for 12 h and monitored the progress of the reaction. The reaction
mixture was cooled to RT and distill off the methanol. The residue was
washed with dry ether and dried under vacuuriz to afford pure magnesium salt
of (-)-3-[4-[2-(2-azo-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid a solid, (weighs about 61 m g, yield : 48.8 %, mp : 230 -
235 °C, purity 90.75 % by HPLC).
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IR (KBr) cm~l : 3425, 2926, 1696, 1624, 1565.
1H NMR (200 MHz, CDC13) 8 : 8.39 (d, J=7.89Hz, 1H), 8.29 (d, J=7.89Hz,
1H), 8.05 - 7.95 (m, 1H), 7.8-7.7 (m, 1H), 7.10 (d, J=8.21Hz, 2H), 6.74 (d,
J=7. 8 9Hz, 2H), 4.4 - 4.3 (m, 4H), 3 . 8 - 3 . 7 (m, 1 H), 3 . 6 - 3 . 5 (m,
1 H), 3 .4 -
5 3.2 (m, 1H), 3.0 - 2.8 (m, 1H), 2.8 - 2.7 (m, 1H), 1.06 (t, J-6:64Hz, 3H).
Example 13
(-)-3-[4-[2-(2-Morpholinyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]
phenyl]-2-ethoxypropanoic acid magnesium salt
0
\ N~° ~ ~ ~_00 M O
~ ~~N~ Et0
10 ~0 2
(-)-3-[4-[2-(2-Morphoinyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-
2-ethoxypropanoic acid (160 mg), dry methanol (5 ml) and magnesium
hydroxide (9.45 mg) was added to 50 ml one necked round bottom flask, fitted
with a mechanical stirrer and reflux condenser. The reaction mixture was
15 refluXed at 80 °C for 12 h and monitored the progress of the
reaction. 'The
reaction mixture was cooled to RT and distill off the methanol. The residue
was washed with dry ether and dried under vacuum to afford pure magnesium
salt of (-)-3-[4-[2-(2-morpholinyl-4-oxo-3,4-dihydroquiriazolin-3-yl)ethoxy]
phenyl]-2-ethoxypropanoic acid a solid, (weighs about 100 rim g, yield : 30.58
20 %, mp : 250 - 255 °C, purity 90.43 % by HPLC).
IR (KBr) cm 1 : 3441, 2924, 2854,1674, 1610, 1587
1H NMR (200 MHz, CDC13) S : 8.16 (d, J=7.89Hz, 1H), 7.71 (d, J=7.06Hz,
1H), 7.60 - 7.50 (m, 1H), 7.50 = 7.35 (m, 1H), 7.16 (d, J=7.98Hz, 2H), 6.75
(d, J=8.21Hz, 2H), 4.56 (t, J = 8. l4Hz, 3H), 4.40 - 4.30 (m, 3H), 3.90 -.
3.65
25 (m, 4H), 3.65 - 3.50 (m, 2H), 3.40 - x.10 (m, SH), 3.05 - 2:90 (m, 1H),
2.90 -
2.70 (m, 1H), 1.15 - 1.00 (m, 3H).
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51
Example 14
(-)-3-[4-[2-(2-Piperidinyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy] phenyl]
-B-ethoxypropanoic acid magnesium salt
O
O +
\ NCO ~ ~ C_OO M O
N ~ N Et0
(-)-3-[4-[2-(2-Piperidinyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxypropanoic acid (147 mg), dry methanol (5 ml) and magnesium
hydroxide (8.72 mg) was added to 50 ml ~ne necked round bottom flask, fitted
with a rilechanical stirrer and reflux condenser. The reaction mixture was
refluxed at 80 °C for 12 h and monitored the progress of the reaction.
The
reaction mixture was cooled to RT and distill off the methanol. The residue
was washed with dry ether and dried under vacuum to afford pure magnesium
salt of (-)-3-[4-[2-(2-morpholinyl-4-oxo-3,4-dihydroquinazolin-3
yl]ethoxy)phenyl]-2-ethoxypropanoic acid as a solid, (weighs about 100 m g,
yield : 33.24 %, mp : 220 - 225 °C, purity 95.30 % by HPLC).
IR (I~Br) cm~l : 2933, 1674, 1610, 1584, 1566.
1H NMIZ (200 MHz, CI~C13) ~ : 8.12 (d, J=7.80Hz, 1H), 7.69 (t, J=4.00Hz,
1H), 7.53 (t, J=8.40Hz, 1H), 7.35 (t, J=7.SOHz, 1H), 7.13 (d, J=8.20Hz, 2H),
6.74 (d, J=8.OOHz, 2H), 4.54 (t, J = 4.90Hz, 2H), 4.31 (t, J=5.20Hz, 2H), 3.90
- 3.75 (m,.1H), 3.65 - 3.50 (m, 1H), 3.33 - 3.26 (m, 1H), 3.20 - 3.03 (m, 4H),
2.95 - 2.70 (m, 2H), 1.75 -1.50 (m, 6H), 1.10 -1.00 (m, 3H).
Example 15
(-)-3-[4-[2-(2-Ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-
ethoxy propanoic acid metformin salt
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52
O -
NH NH
O ~~ II
NCO ~ ~ C_O ~. ~ N~N~N.Me
3
Et0 H Me
(-)-2-EthoXy-3-[4-[2-(2-ethyl-4-axo-3,4-dihydro-3-quinazolinyl)ethoxy]
phenyl]propionic acid (2.0 g) and isopropanol (50 ml) was added to 250 ml
four necked round bottom flask, fitted with a mechanical stirrer and reflux
condenser. The reaction mixture was slowly heated to 45-55° for
complete
dissolution of the mass. Metformin (0.663 g).dissolved in isopropanol (10 ml)
was .added to the reaction mixture of 75-85 °C for 5 h and monitor the
progress of the reaction. The reaction mixture was cooled to room temperature
and stirred for 12 h at room temperature. Distill off the isopropanol on
rotavapor water bath at 45-55 °C under vacuum. The precipitated product
was
filtered, dried at 60 °C for 2-3 h to afford the pure metformin salt of
(-)-3-[4-
[2-(2-ethyl-4-oxo-3,4-dihydroquinazolin-3-yl)ethoxy]phenyl]-2-ethoxy
propanoic acid as off white crystalline solid, (weighs about 2.0 g, Yield : 77
%, m.p. 164-66 °C, purity 99% by HPLC).
IR as KBr shows the following absorption bands (cm 1) 3400-3300 (N-H
stretch), 3120 (-C-H aromatic), 2930 (-C-H aliphatic), 1660 (-COO stretch),
1587. (-CONH stretch), 1400 (-COO stretch).
1H NMR spectrum in I7MS0-d6 (TMS as internal standard) shows the
following signals 8 8.13 (d, J=7.89Hz, 1H), 7.82 (t, ~J=7.01Hz, 1H), 7.64(d,
J=8.21Hz, 1H), 7.50 (t, J= 7.26Hz, 2H), 7.13(d, J= 8.SOHz, 2H), 684 (d,
J=B.SOHz, 2H), 4.47 (t, J=5.19Hz, 2H), 4.26 (t, J=5.19Hz, 2H), 3.99-3.84 (m,
1H), 3.60=3.40 (m, 1H), 3.40-3.20 (m, 1H), 3.06 (q, J= 6.96Hz, 2H), 2.88 (q,
J= 6.64Hz, 2H), 2.8 (s, 6H), 1.32 (t, J= 7.17Hz, 3H), 1.02 (t, J= 6.96Hz, 3H).
The mass spectrum shows m/z, 411 (M+ + 1), 130 (C4H11N5), Anal. Calcd :
C23H2gNaO5; % C 60.11; % H : 6.86; % N 18.18; Found % C 59.89; % H 6.66;
N 18.00.
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Example-lfi
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin=6-yl)ethoxy]phenyl]-2-ethoxypropionic acid magnesium salt
O _
NCO ~ ~ ~
C-O
N ~ I N Et0 Mg++
2
(-) ' 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimi'din-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), methanol (50
ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and . reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Magnesium
hydroxide (0.328 g) was added to the reaction mixture at 60 °C in about
10
minutes under stirring. Maintained gentle reflux of the reaction mixture for
12-
14 hr and monitored the progress of the reaction. The reaction mixture was
cooled t~ RT and stirred for 2-3 h at room temperature. The precipitated
product was filtered, vacuum dried to afford the pure magnesium salt of (-) 3-
[4-[2-(1,5-dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white
crystalline solid, (weighs about 4.09 g, yield : 80 %, rrip 113 °C,
purity 99
by HPLC).
IR (KBr) cm 1 : 3120 (C-H aromatic), 2930 (-C-H aliphatic), 1693 (-COO / -
CONH stretch), 1574 (-CONH amide-II band).
1H NMR (200 MHz, l~MSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2Ii), 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, T= 4.6 Hz, 2H), 4.35-4.15 (m, SH), 4.01-3.97 (m, 1H), 3.56-3.38
(m, 2H), 3.12-2.90 (in, 2H), 2.83 (t; J=7. Hz, 2H), 2.76 (s, 3H), 1.89-1.71
(m,
2H), 1.16 (t, J= 7.0 Hz, 3H), 0.98 (t, J= 7.3 Hz, 3H).
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Mass m/z : 443 (M+ + 1), Anal. Calcd : C46HS8NsOloMg; % C 60.92; % H
6.40; % N 12.31; Found % C 60.80; % H 6.30; % N 12.20.
Example-17
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid potassium salt
O _
O ~ / ~ ~ O
-C-O K
Et0
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), methanol (50
ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Potassium
hydroxide
(0.63 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to
1ZT and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vac~ium- dried to afford the pure potassium salt of (-) 3-[4-[2-(1,5-
dimethyl-7-oxo-3-propyl-6, 7-dihydro=1 H-pyrazolo [4, 3-d]pyrimidin-6-
yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white crystalline solid
(weighs about 4.6 g, yield : 85 %, mp : 168-170 °C, purity 99 % by
HPL,C).
IR (KBr) cm-1 : 3120 (C-H aromatic), 2960 (-C-H aliphatic), 1690 (-COO I -
CONK stretch), 1573 (-CONH amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H); 6.78 (d, J=8.3 Iiz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, SH), 4.01-3.97 (m, 1H), 3.56-3.38
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(m, 2H), 3.12-2.90 (m, 2H), 2.83 (t, J~7.8 Hz, 2H), 2.76 (s, 3H), 1.89-1.71
(m,
2H), 1.16 (t, J= 7.0 Hz, 3H), 0.98 (t, J= 7.3 Hz, 3H).
Mass mlz : 443 (M+ + 1), Anal. Calcd : Ca3Hz9N~OsK; % C 57.49; % H : 6.04;
N 11.66; Found % C 57.30; % H 5.9; % N 11.50.
5
Example-18
(-) 3-[4-[2-(1;5=Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-B-ethoxypropionic acid calcium salt
~ _
NCO ~ ~ 101 O
N ~ .
\ N~ Ca++
Et0
/ 2
10 (-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimid~z-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), methanol (50
ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Calcium
hydroxide
15 (0.418 g) was added to the reaction mixture at 60 °C in about 10
minutes
under stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr
and monitored the progress of the reaction. The reaction mixture was cooled ,
to RT and stirred for 2-3 h at room temperature. The precipitated product Was
filtered, vacuum- dried to afford the pure calcium salt of (-) 3-[4-[2-(1,5-
20 dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-6-
yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white crystalline solid
(weighs about 4.17 g, yield : 80 %, mp : 251 °C (dec), purity 99 % by
HPLC).
IR (KBr) cm 1 : 3150 (C-H aromatic), 2960 (-C-H aliphatic), 1690 (-COO / -
CONH stretch), 1577 (-CONH amide II band).
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56
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H), 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, SH), 4.10-3.97 (m; 1H), 3.56-3.38
(m~ 2H), 3.12-2.90~(m, 2H), 2.83 (t, J=7.8 Hz, 2H), 2.76 (s, 3H), 1.89-1.71
(m,
2H), 1.16 (t, J= 7.0 Hz, 3H), 0.98 (t, J= 7.3 Hz, 3H).
Mass m/z : 443 (M+ + 1), Anal. Calcd : C46HssN8oioCa; % C 59.86; % H :
6.29; % 1V~ 12.14; Found % C 59.70; % H 6.15; % N 12Ø
Example-19
(-) , 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lithium salt
O
NOD
Li
N ~ I ~ Et
-N
3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidiiz-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), methanol (50
ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Lithium
hydroxide
(0.47 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum- dried to afford the pure lithium salt of (-) 3-[4-[2-(1,5-
dimethyl-7-oxo-3-propyl-6,7-dihydro-1 H-pyrazolo [4,3-d]pyrimidin-6-
yl)ethoxy]phenyl]-2-ethoXypropionic acid as off white crystalline solid
(weighs about 4.3 g, yield : 85 %, mp : 254-266 °C, purity 99 % by
HPLC).
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IR (KBr) cm 1 : 3150 (C-H aromatic), 2950 (-C-H aliphatic), 1690 (-COO / -
CONH stretch), 1574 (-CONH amide-II band).
1H NMIZ (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H), 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, SH), 4.10-3.97 (m, 1H), 3.56-3.38
(m, 2H), 3.12-2.90 (m, 2H), 2.83 (t, J=7.8 Hz, 2H), 2.76 (s, 3H), 1.89-1.71
(m,
2H), 1.16 (t, J= 7,0 Hz, 3H), 0.98 (t, J= 7.3 Hz, 3H).
Mass m/z ~ : 443 (1VI+ + 1), Anal. Calcd : Ca3H29N4OsLi; % C 61.60; % H
6.47;%N12.49;Found%C61.45;%H6.31;%N12.35.
Example-20
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoXypropionic acid sodium salt
O _
NCO
N ~ ,--C-O Na
Et0
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), methanol (50
ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Sodium
hydroxide
(0.45 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum- dried to afford the pure sodium salt of (-) 3-[4-[2-(1,5-
dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-6-
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58
yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white hygroscopic solid
(weighs about 4.19 g, yield : 80 %, purity 99 % by HP1,C).
IR (I~Br) cm 1 : 3100 (C-H aromatic), 2963 (-C-H aliphatic), 1688 (-COO / - ,
CONH stretch), 1574 (-CONH amide-II band).
IH NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H), 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, 5H), 4.10-3.97 (m, 1H), 3.56-3.38
(m, 2H), 3.12-2.90 (m, 2H), 2.83 (t, J=7.8 Hz, 2H), 2.76 (s, 3H), 1.89-1.71
(m,
2H), 1.16 (t, J= 7.0 Hz, 3H), 0.981(t, J= 7.3 Hz, 3H).
Mass m/z : 443 (M++ 1).
Example-21
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid arginine salt
O
NCO \-/ O O H NH2
C-O HN N
N ~ I ~ Et ~ H C02H
N ~NH3
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 riml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. 1;-Argiriine
(1.96 g)
was added to the reaction mixture at 60° in about 10 minutes under
stirring.
Maintained gentle reflux of the reaction mixture for 12-14 hr and monitored
the progress of the reaction. The reaction mixture was cooled to RT and
stirred
for 2-3 h at room temperature. The precipitated product was filtered, vacuum-
dried to afford the pure arginine salt of (-) 3-[4-[2-(1;5=dimethyl-7-oxo-3-
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59
propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-6-yl)ethoxy]phenyl]-2-
ethoxypropionic acid as off white crystalline solid (weighs about 5.92 g,
yield
85 %, mp : 192 °C, purity 99 % by HPLC).
IR (KBr) cm 1 : 3300-3200 (-NH stretch), 3150 (C-H aromatic), 2950 (-C-H
aliphatic), 1690 (-COO / -CONH stretch), 1583 (-CONH amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H), 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, SH), 4.10-3.97 (m, 1H), 3.80 (m,
2H), 3.56=3.38 (m, 2H), 3.20 (m, 2H), 3.12-2.90 (m, 2H), 2.83 (t, J=7.8 Hz,
2H), 2.76 (s, 3H), 2.0-1.6 (m, 3H), 1.89-1.71 (m, 2H), 1.16 (t, J= 7.0 Hz,
3H),
0.98 (t, J= 7.3 Hz, 3H).
Mass xn/z : 443 (1VI+ + 1), Anal. Calcd : C2~H44N807i % ~ 56.49; % H : 7.14;
N 18.18; Found % C 56.34; % H 6.98; % N 17.99.
Example-22
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid 1t-(+) methyl
benzylamine salt
,. ~ O
N ~C - C O NH3
N \ ( N ~ ~ C_O \ CHs
EtO
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro=1H-pyrazolo[4,3-
d]pyiimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. R-(+) methyl
benzylamine (1.36 g) was added to the reaction mixture at 60 °C in
about 10
minutes under stirring. Maintained gentle reflux of the reaction mixture for
12-
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- 14 hr and monitored the progress of the reaction. The reaction mixture vvas
cooled to RT and stirred for 2-3 h at room temperature. The precipitated
product was filtered, vacuum- dried to afford the pure, hygroscopic (-) 3-[4-
[2-
(1,5-dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-6-
5 yl)ethoxy]phenyl]-2-ethoxypropionic acid R-(+) methyl benzylamine salt
as off white crystallixie solid (weighs about 5.41 g, yield : 85 %, mp : 114-
115
°C~ pm'itY 99 % by HPI,C).
IR (KBr) crri 1 : 3150 (C-H aromatic), 2940 (-C-H aliphatic), 1687 (-COO / -
CONH stretch), 1573 (-CONH amide-II band).
10 1H NMR (200 MHz, DMSO-d6) 8 : 7.40-7.20 (m, 5H), 7.13 (d, J=8.3 Hz, 2H),
6.78 (d, J=8.3 Hz, 2H), 4.48 (t, J= 4.6 Hz,. 2H), 4.35-4.15 (m, SH), 4.10 (m,
1H), 4.10-3.97 (m, 1H), 3.56-3.38 (m, 2H), 3.12-2.90 (m, 2H), 2.83 (t, J=7.8
Hz, 2H), 2.76 (s, 3H), 1.89-1.71 (m, 2H), 1.40 (d, 3H), 1.16 (t, J= 7.0 Hz,
3H),
0.98 (t, J= 7.3 Hz, 3H). ,
15 Mass m/z : 443 (M+ + 1), tonal. Calcd : C31H41NSO5; % C 66.07; % H : 7.28;
N 12.43; Found % C 65.90; % H 7.15; % N 12.33.
Example-23
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H=pyrazolo(4,3-
20 d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid S-(+)-
~phenylglycinol salt
O Ph
N~~ ~ / ~ ~
N ~ C-O ~3N~OH
Et0
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo=3-propyl-6,7-dihydro-1H-pyrazolo[4,3
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
25 (50 ml) ~,vere added to 250 ml four necked round bottom flask, fitted with
a
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mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. S-(+)-
phenylglycinol
(1.549 g) was added to the reaction mixture at 60 °C in about 10
minutes
under stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr
and monitored the progress of the reaction. The reaction mixture was cooled
to RT and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure S-(+)=phenylglycinol salt of (-) 3-
[4-
[2-( 1, 5-dimethyl-7-oxo-3-propyl-6, 7-dihydro-1 H-pyrazolo [4, 3-d]pyrimidin-
6-
yl)ethoxy]phenyl]-2-ethoxypropionic acid of the formula as off white
hygroscopic solid (weighs about 5.23 g, yield : 80 %, purity 99
°l° by HPLC).
IR (KBr) cm 1 : 3400-3300 (O-H stretch), 3150 (C-H aromatic), 2950 (-C-H
aliphatic), 1689 (-COO / -CONH stretch), 1573 (-CONH amide-II band).
1H NMR (200 MHz, I~MSO-d6) ~ : 7.40 (m, SH), 7.13 (d, J=8.3 Hz, 2H), 6.78
(d, J=8.3 Hz, 2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, SH), 4.10-3.97 (m,
1H), 4.00 (d, 2H), 3.60-3.50 (m, 1H), 3.56-3.38 (m, 2H), 3.12-2.90 (m, 2H),
2.83 (t, J=7.8 Hz, 2H), 2.76 (s, 3H), 1.89-1.71 (m, 2H), 1.16 (t, J= 7.0 Hz,
3H),
0.98 (t, J= 7.3 Hz, 3H).
Mass m/z : 443 (M+ + 1).
Example-24
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminogeaanidine
hydrogen carbonate salt
O NW
NCO ~ ~ ~ ~ ,~ HCOO~
N ~ rC-O H3N N 3
Et0 Q H
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(-) 3-[4-[2-(1,5-I7imethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionie acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, gifted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °c for complete dissolution of the . mass. 1.53 g of
Aminoguanidine hydrogen carbonate°was added to the reaction
mixture at 60
°C in about 10 minutes under stirring. Maintained gentle reflux of the
reaction
mixture for 12-14 hr and monitored the progress of the reaction. The reaction
mixture was cooled to RT and stirred for 2-3 h at room temperature. The
precipitated product was filtered, vacuum- dried to afford the pure
aminoguanidine salt of (-) 3-[4-[2-(1,5-diinethyl-7-oxo-3-propyl-6,7-dihydro-
1H-pyrazolo[4,3-d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid as
off white hygroscope solid, (weighs about 5.5 g, yield : 85 %, mp : 110-112
°C, purity 99 % by Hl'LC).
IR (I~r) cm' : 3450-3350 (-NH stretch), 3150 (C-H aromatic), 2956 (-C-H
aliphatic), 1684 (-COO / -CONH stret'ch), 1572 (-CONH amide=II band).
1~I NMR (200~MHz, I~MSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H), 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, SH), 4.10-3.97 (m, 1H), 3.56-3.38
(m, 2H), 3.12-2.90 (W , 2H), 2.83 (t, J=7.8 Hz, 2H), 2.76 (s, 3H), 1.89-1.71
(m,
2H), 1.16 (t, J= 7.0 Hz, 3H), 0.98 (t, J= 7.3 Hz, 3H).
Mass m/z : 443 (M+ + 1).
Example-25
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo(4,3
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid tromethamine
s alt
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O OH
NCO ~ ~ ~ OH
rc_O
N ~ I N ~ Et0 N3N OH
(-) . 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stiirer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Trdmethamine
(1.36
g) was added to the reaction mixture at 60 °C in about 10 minutes undei
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled,to
RT and stirred for 2-3 h at room temperature. The precipitated product was
filtered, aacuum- dried to afford the pure tromethamine salt of (-) 3-[4-[2-
(1,5-
dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-6-
yl)ethoxy]phenyl]-2-ethoxypropionic ~ acid as off white hygroscopic solid
(weighs about 5.1 g, yield : 80 %, purity 99 % by HPLC).
IR (KBr) cmi 1 : 3300-3250 (O-H stretch), 3120 (C-Ii aromatic), 2950 (-C-H
aliphatic), 1689 (-COO / -CONH stretch), 1574 (-CONH, atiiide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H); 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, SH), 4.10-3.97 (m, 1H), 4.00 (s,
6H), 3.56-3.38 (m, 2H), 3.12-2.90 (m, 2H), 2.83 (t, J=7.8 Hz, 2H), 2.76 (s,
3H), 1.89-1.71 (m, 2H), 1.16 (t, J= 7.0 Hz, 3H), 0.98 (t, J= 7.3 Hz, 3H).
Mass m/z : 443 (M+ + 1).
Example-26
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(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4;3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid
dicyclohexylamine salt
O _
O ~N2
C-O
EtO
(-)-3-[4-[2-(1,5-I~imethyl)]-7-oxo-3-propyl-6,7-diliydro-1H-pyrazolo[4,3-d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass.
Dicyclohexylamine
(2.04 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to RT
and 'stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure dicyclohexylamine salt of (-) 3-[4-
[2-
(1,5-dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3=d]pyrimidin-6-
yl)ethoxy]phenyl]-2-ethoxypropionic acid as . off white crystalline solid
(weighs about 5.6 g, yield : 80 %, mp : 132-134 °C, purity 99 % by
HPLC).
IR (KBr) cm 1 : 3150 (C-H aromatic), 2950 (-C-H aliphatic), 1690 (-COO / -
CONH stretch), 1574 .(-CONH amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H), 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, 5H), 4.10-3.97 (rri, 1H), 3.56-
3.38
(m, 2H), 3.12-2.90 (m, 2H), 2.83 (t, J=7.8 Hz, 2H), 2.76 (s, 3H), 2.60-1.00
(m,
22H)~, 1.89-1.71 (m, 2H), 1.16 (t, J= 7:0 Hz, 3H), 0.98 (t, J= 7.3 Hz, 3H).
Mass m/z : 443 (M++ 1), Anal. Calcd : C35Hs3NsOs; % C 67.41; % H : 8.5;
N 11.23; Found % C 67.25; % H 8.4; % N 11.15. ~.,
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Example-27
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-
5 octylglucamine salt
O CH2-NH2-C$H~~
NCO ~ ~ 101
Etp C O H OH
N HO H
H OH
H OH
CH20W
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidill-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
10 mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. N-
octylglucamine
(3.31 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 h and
monitored the progress of the reaction. 'fhe reaction mixture was cooled to
15 RT and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure N-octylglucamine salt of (-) 3-[4-[2-
(1,5-dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-6-
yl)etlioxy]phenyl]-2-ethoxypropionic acid as off white hygroscopic solid
(weighs about 6.6 g, yield : 80 %, purity 9'9 % by HPLC).
20 IR (KBr) cm 1 : 3350-3300 (O-H stretch), 3150 (C-H aromatic), 2930 (-C-H
aliphatic), 1689 (-COO / -CONH stretch), 1576 (-CONH amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H)~ 6.78 (d, J=8.3 Hz,
2H), .4.48 .(t, J= 4.6 Hz, 2H), 4.35-4.1 S (m, SH), 4.20-3.01 (m, 8H), 4.01
(m,
1H), 4.10-3.97 (m, 1H), 3.56-3.38 (m,, 2H), 3.12-2.90 (rri, 2H), 2.83 (t,
J=7.8
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Hz, 2H), 2.76 (s, 3H), 1.89-1.71 (m, 2H), 1.42 -1.00 (m, 16H), 1.16 (t, J= 7.0
Hz, 3H), 0.98 (t, J= 7.3 Hz, 3H).
Mass m/z : 443 (M+ + 1).
Example-28
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)'ethoxy]phenyl]-2-ethoxypropionic acid N-
methylglucamine salt
O C H2-iV H2-C H3
N ~ ~ ~ O ~
H ' OH
C O
N~ Et0 H~ H
~. OH
H OH
CH20H
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro=1H-pyrazolo[4;3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. 'The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. N-
inethylglucamine
(2.2 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum- dried to afford the pure, hygroscopic N-methylglucamine
salt of (-) 3=[4-[2-(1,5-dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4;3-
d]pyrirriidin-6-yl)ethoXy]phenyl]-2-ethoxypropionic acid as off white
crystalline solid (weighs about 6.1 g, yield : 85 %, inp 115 °C, purity
99 % by
HPLC).
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IR (KBr) cm 1 : 3400-3200 (-OH stretch), 3150 (C-H aromatic), 2950 (-C-H
aliphatic), 1680 (-COO / -CONH stretch), 1 S74 (-CONH amide-TI band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H), 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, SH), 4.10-3.97 (m, 1H), 4.00-3.26
S (m, 8H), 3.56-3.38 (m, 2H), 3.12-2.90 (m, 2H), 2.83 (t, J=7. Hz, 2H), 2.76
(s,
3H), 2.40 (s, 3H), 1.89-1.71 (m, 2H), 1.16 (t, J= 7,0 Hz; 3H), 0.98 (t, J= 7.3
Hz, 3H).
Mass m/z : 443 (M+ + 1), Anal. Calcd : C3oH47NsOlo; % C S6.S; % H : 7.37;
N 10.98; Found % C S6.3S; % H 7:25; % N 10.8.
Example-29
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-B-ethoxypropionic acid metformin salt
O NH NH
N NCO \ / 101 ~ ~ N~N~N.NIe
N , ~-c-~
Et0 H Me
/ .
1S (-) ' 3-[4-[2-(1,S-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isoproparol
(SO ml) were added to 2S0 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to SS-60 °C for complete dissolution of the mass. Metformin free
base
(1.45 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was 'cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure metformin salt of (-) 3-[4-[2-(1,S-
2S dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3=d]pyrimidin-6-
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yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white hygroscopic solid
(weighs about 5.16g, yield : 80 %, purity 99 % by HPLC).
IR (KBr) cm 1 : 3400-3300 (N-H stretch), 3190 (C-H aromatic), 2968 (-C-H
aliphatic), '1687 (-COO / -CONH stretch), 1573 (-CONH amide-II band).
1H NMI~ (200 MHz, DMSO-d~) 8 : 7.13 (d, J=8.3 I-~z, 2H), 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, 5~I), 4.10-3.97 (rim, 1H), 3.56-
3.38
(m, 2H), 3.12-2.90 (m, 2H), 2.90 (s, 6H), 2.83 (t, 3=7.8 Hz, 2H), 2.76 (s,
3H),
1.89-1.71 (m, 2H), 1.16 (t, J= 7.0 Hz, 3H), 0.98 (t, J= 7.3 Hz, 3H).
Mass m/z : 443 (M++ 1).
example-30
(-) 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lysine salt
O O
NCO ~ l o ~ NH3
N ~ I . i Et ~C-O ~ (CH2)a.
N
H--~-N H2
COON
(-) , 3-[4-[2-(1,5-Dimethyl-7-oxo-3-propyl-6,7-dihydr'o-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Lysine
monohydrate
free base (1.65 g) was added to the reaction mixture at 60 °C in about
10
minutes under stirring. Maintained gentle reflux of the reaction mixture for
12-
14 hr and monitored the progress of the reaction. The reaction mixture was
cooled to RT and stirred for 2-3 h at room temperature. The precipitated
product was filtered, vacuum dried to afford the pure lysine salt of (-) 3-[4-
[2-
(1,5-dimethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-6-
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yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white crystalline solid
(weighs about 5.98 g, yield : 90 %, mp : 158-160 °C, purity 99 % by
HPLC).
IR (I~r) cm 1 : 3400-3300 (N-H stretch), 3150 (C-H aroW atic), 2950 (-C-H
aliphatic), 1694 (-COO / -CONH stretch), 1573 (-CONH amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.3 Hz, 2H), 6.78 (d, J=8.3 Hz,
2H), 4.48 (t, J= 4.6 Hz, 2H), 4.35-4.15 (m, SH), 4.20 (t, 1H); 4.1-3.97 (m,
1H),
3.56-3.38 (m, 2H), 3.12-2.90 (rim, 2H), 3.01 (t, 2H), 2.83 (t, J=7.8 Hz, 2H),
2.76 (s, 3H), 2.20-1.5 (m, 6H), 1.89-1.71 (m, 2H), 1.16 (t, J= 7.0 Hz, 3H),
0.98
(t, J= 7.3 Hz, 3H).
Mass m/z : 443 (M+ + 1), Anal. Calcd : C29H44N6O7; % C 59.18; % H : 7.46;
N 14.78; Found % C 59.0; % H 7.29; % N 14.59:
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-lI-I-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid was prepared
according .to the procedure described in our copending IJS patent application
No. 09/507,373
[2S, N(1S)]-2-Ethoxy-3-[4-[2-(5-ethyl-1-methyl-7-oxo-3-propyl-6,7-dihydro-
1 H-pyrazolo [4, 3-d]pyrimidin-6-y] ethoxy]phenyl]-N-(2-hydroxy-1-
phenylethyl)propanamide was taken, in a mixture of dioxarie, water and 1M
sulfuric acid at room temperature and stirred under reflux for 34 h. Water and
dioxane were removed under vacuum. The residue was taken in water and
extracted with ethyl acetate and the ethyl acetate layer was washed with
water,
dried (Na2S04) and evaporated to dryness to yield the crude compound. The
crude compound was purified by column chromatography using 50 % ethyl
acetate in pet. ether as an eluent to afford (-)-3-[4-[2-(1-methyl-5-ethyl-7-
oxo-
3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-6-yl)ethoxy]phenyl]-2- .
ethoxypropionic acid.
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However, any other procedure for preparing (-)-3-[4-[2-(1-methyl-5-
ethyl-7-oxo-3-propyl-6,7-dihydro-1 H-pyrazolo [4, 3-d]pyrimidin-6-
yl)ethoxy]phenyl]-2-ethoxypropionic acid can be used. (~) 3-[4-[2-(1-methyl-
5-ethyl-7-oxo-3-propyl-6, 7-dihydro-1 H-pyrazolo [4, 3-d]pyrimidin-6-yl)
5 ethoxy]phenyl]-2-ethoxypropionic acid and (+)-3-[4-[2-(1-methyl-5-ethyl-7-
oxo-3-propyl-6,7-dihydro-1H-pyrazolo[~.,3-d]pyrimidin-6-yl)ethoxy]phenyl]-
2-ethoxypropionic acid can be prepared by a similar procedure described
above.
10 Example-31
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo [4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid magnesium salt
O
C-O
Et0 Mg++
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]
15 pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), methanol (50
ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass and magnesium
hydroxide (0.29 g) was added to the reaction mixture at 60 °C in about
10
20 minutes under stirring. Maintained gentle reflux of the reaction mixture
for 12-
14 hr and monitored the progress of the reaction. The reaction mixture was
cooled to ' RT and stirred for 2-3 h at room temperature. The precipitated
product was filtered, vacuum dried to afford the pure magnesium salt of (-)-3-
[4-[2-(1-methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3- .
25 d]pyrimidin-6-yl)ethoxy] phenyl]-2-ethoxypropionic acid as off white
crystalline solid, (weighs about 4.0 g, yield : 80 %, purity f9 % by HPLC). .
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IR (KBr) crri 1 : 3120 (C-H aromatic), 2970 (-C-T~ aliphatic), 1688 (-COO l -
CONH stretch), 1576 (amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.4 Hz, 2H), 6.77 (d, J=8.6 Hz,
2H), 4.50 (t, J= 5.0 Hz, 2H), 4.29-4.22 (m, 5H), 4.01 (t, J=3.7 Hz, 1H), 3.69-
~ 3.35 (m, 2H), 3.10-2.94 (m, 4H), 2.84 (t, J=7.5 Hz, 2H), 1.90-1.78 (m, 2H),
1.30 (t, J= 7.0 Hz, 3H), 1.16 (t, J=7.4 Hz, 3H), 0.97 (t, J= 7.4 Hz, 3H).
Mass m/z : 457 (M+ + 1), Anal. Calcd : C4gH6zN8~IOMg~ % C 61.67; % H :
6.63; % N 11.99; Found % C 61.58; % H 6.55; % N 11.84.
Example-3B
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7=dihydro-1~I-pyrazolo [4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid potassium salt
O
O
-C-O K
Et0
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1I-I-pyrazolo[4,3-d]
15. pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Potassium
hydroxide
(0.61 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure potassium salt of (-)-3-[4-[2-(1-
methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d] pyrimidin-6-
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yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white ainoiphous solid,
(weighs about 4.35 g, yield : 80 %, mp : 164 °C, purity 99 % by HPLC).
IR (It.Br) cm 1 : 3120 (C-H aromatic), 2970 (-C-H aliphatic), 1681 (-COO / -
C~NH stretch), 1576 (amide-II band).
1H NMR (200 MHz, DMSO-d6) ~ : 7.13 (d, J=8.4 Hz, 2H), 6.77 (d, J=8.6 Hz,
2H), 4.50 ,(t, J= ~.0 Hz, 2H), 4.29-4.22 (rim, SH),-4.01 (t, J=3.7 Hz, 1H),
3.69-
3.35 (m, 2H), 3.10-2.94 (m, 4H), 2.84 (t, J=7.5 Hz, 2H), 1.90-1.78 (m, 2H),
138 (t, J= 7.3 Hz, 3H), 1.16 (t, J=7.0 Hz, 3H), 0.97 (t, J= 7.4 Iiz, 3H).
Mass spectrum shows m/z : 457 (M+' + 1), Anal. Calcd : Ca4H31N405R~
°a C
58.29; % H : 6.27; % N 11.33; Found % C 58.1; % H 6.15; % N 11.20.
lJxariiple-33 .
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo [4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid calcium salt
. N NBC \ .~ ~ ~
C-O
Et0 Ca++
2
(-)-3-[4-[2-( 1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1 H-pyrazolo[4,3-d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), methanol (50
ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and ~ reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Calcium
hydroxide
(0.81 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
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73
filtered, vacuum dried to afford the pure calcium salt of (-)-3-[4-[2-(1-
Methyl-
5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d] pyrimidin-6-
yl)ethoxy]phenyl]-2-ethoxypropionic ~ acid as off White solid, (weighs about
4.4 g, yield : 85 %, mp : >260 °C, purity 99 % by HPLC).
IR (KBr) cm 1 : 3120 (C-H aromatic), 2970 (-C-H aliphatic), 1681 (-COO l -
CONH stretch), 1576 (-CONH stretch).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.4 Hz, 2H), 6.77 (d, J=8.6 Hz,
2H), 4.50 (t, J= 5.0 Hz, 2H), 4.29-4.22 (m, SH), 4.01 (t, J=3.7 Hz, 1H), 3.69-
3.35 (m, 2H), 3.1-2.94 (m, 4H), 2.84 (t, J=7.3 Hz, 3H), 1.90-1.78 (m, 2H),
1.38 (t, J= 7.3 Hz, 3H), 1.16 (t, J=7.0 Hz, 3H), 0.97 (t, J= 7.4 IIz, 3H).
Mass m/z : 457 (M+ + 1), Anal. Calcd : C48H62N$OloCa; % C 60.63; % H
6.52; % N 11.78; Found % C 60.51; % H 6.4; % N 11.60.
Example-34
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid lithium salt
O
N NCO \ . / 101. O
N ~ ,--O_~ a
Et0
i
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoXypropionic acid (5.0 g), methanol (50
ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Lithium
hydroxide
(0.44 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to RT
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and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure lithium salt of (-)-3-[4-[2-(1-
methyl-
5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d] pyrimidin-6-
yl)ethoxy]phenyl]-2-ethoxypropionic acid as ~ off white amorphous solid,
(weighs about 4.55 g, yield : 90 %, mp : 215-218 °C, purity 99% by
HPLC).
IR (KBr) cm 1 : 3120 (C-H aromatic), 2857 (-C-H aliphatic), 1686 (-COO / -
CONH stretch), 1570 (amide-II band).
1H NMR (200 MHz, I7MSO-d6) 8 : 7.13 (d, J=8.4 Hz, 2H), 6.77 (d, J=8.6 Hz,
2H), 4.50 (t, J= 5.0 Hz, 2IY), 4.29-4.22 (m, SH), 4:01 (t, J=3.7 Hz, lI~),
3.69
3.35 (m, 2H), 3.1-2.94 (m, 4H), 2.84 (t, J=7.5 Hz, 2H), 1.90-1.78 (m, 2H),
1.38 (t, J= 7.3 Hz, 3H), 1.16 (t, J=7.OHz, 2H), 0.9'7 (t, J= 7.4 Hz, 3H).
Mass m/z : 457 (M+ + 1), Anal. Calcd : C24H3iN40sLi; % C 62.33; % H :
6.70;. % N 12.12; Found % C 62.1; % H 6.58; % N 12.10
Example-35
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo (4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid sodium salt
O
0 \ / ~ ~ N~ ._
C-O
Ef0
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), methanol (50
ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Sodium
hydroxide
(0.43 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
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monitored the progress of the reaction. The reaction mixture was cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure sodium salt of (-)-3-[4-[2-(1-methyl-
5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d] pyrimidin-6-
5 yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white amorphous solid,
(weighs about 4.25 g, yield : 80 %, mp : 112-114 °G, purity 99 % by
HPLC).
IR (I~.Br) cm 1 : 3120 (C-H aromatic), 2970 (-C-H aliphatic), 1680 (-COO / -
CONH stretch), 1570 (amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.4 Hz, 2H); 6.77 (d, J=8.6 Hz,
10 2H), 4.50 (t, J= 5.0 Hz, 2H), 4.29-4.22 (m, SH), 4.01 (t, J=3.7 Hz, 1~I),
3.69-
3.35 (rim; 2H), 3.10-2.94 (m, 4H), 2.84 (t, J=7.5 Hz, 2H), 1.90-1.78 (m; 2H),
1.16 (t, J= 7.0 Hz, 3H), 1.38 (t, J= 7.3 Hz, 3H), 0.97 (t, J= 7.4 Hz, 3H).
Mass mlz : 457 (M+ + 1), Anal. Calcd : Ca4H31N4O5Na; % C 60.25; % H
6.48;%N11.71;Found%C60.15;°/.°H6.31;%N11.58.
Example=36
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H=pyrazolo [4,3=
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid arginine salt
p _
N~~ ~ I O ~ HN N NH2
N ~ ( N ~ Et~ C O H C~2H
~NH3
(-)-3-[4-[2-( 1-Methyl-5-ethyl-7-oxo-3 -propyl-6,7-dihydro-1 H-pyrazolo [4, 3-
d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic . acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the rr~ass. L-Arginine
(1.9 g)
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was added to the reaction mixture at 60 °C in about 10 minutes under
stirring.
Maintained gentle reflux of the reaction mixture for 12-14 hr and monitored
the progress of the reaction. The reaction mixture was cooled to RT and
stirred
for 2-3 h at room temperature. The precipitated product was filtered, vacuum
dried to afford the pure arginine salt of (-)-3-[4-[2-(1-methyl-5-ethyl-7-oxo-
3-
propyl-6;7-dihydro-1H-pyrazolo[4,3-d] pyrimidin-6-yl)ethoxy]phenyl]-2
ethoxypropionic acid as off white amorphous solid, (weighs about 5.5 g, yield
80 %, mp : 235 °C, purity 99 % by HPLC).
IR (KBr) , cm 1 : 3360 (-NH stretch), 3120 (C-H aromatic), 2957 (-C-H
aliphatic), 1688 (-COO / -CONH stretch), 1573 (amide-II band).
1H NMR (200 MHz, I~MSO-d6) 8 : 7.13 (d, J=8.4 Hz; 2H),'6.77 (d, J=8.6 Hz,
2H), 4.50 (t, J= 5.0 Hz, 2H), 4.29-4.22 (m, SH), 4.01 (t, J=3.7 Hz, 1H), 3.80
(m, 2H), 3.69-3.35 (m, 2H), 3.20 (m, 3H), 3.10-2.94 (m, 4H), 2.84 (t, J=7.5
Hz, 2H), 1.90-1.78 (m, 2H), 1.60-2.00 (m, 3H), 1.38 (t, J= 7.3 Hz, 3H), 1.16
(t,
J= 7.0 Hz, 3H), 0.97 (t, J= 7.4 Hz, 3H).
Mass m/z : 457 (M+ + 1), Anal. Calcd : C3oH4gN807; % C 57.14; % H : 7.3;
N17.75;Found%C57;%H7.15;%N17.58.
Example-37
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid R-(+) methyl
benzylamine salt
O
O ~ ~ ~ 0 NH3
C-O
~EtO ~
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydio-1H-pyrazolo[4,3-d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
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(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. R-(+) methyl
benzylamilie (1.32 g) was added to the reaction mixture at 60 °C in
about 10
minutes under stirring. Maintained gentle reflux of the reaction mixture for
12-
14 hr and monitored the progress of the reaction. The reaction mixture was
cooled to RT and stirred for 2-3 h at room temperature. The precipitated
product was filtered, vacuum dried to afford the pure, hygroscopic (-)-3-[4-[2-
( 1-methyl-5-ethyl-7-oxo-3-propyl-6, 7-dihydro-1 H-pyrazolo [4, 3 -
d]pyrimidin~
6-yl)ethoxy] phenyl]-2-ethoxypropionic acid R-(+) methyl benzylamine salt
as off white solid, (weighs about 4.4 g, yield : 70 %, mp : 164-166 °C,
purity
99 % by HPLC).
IR (KDr) cm 1 : 3120 (C-H aromatic), 2970 (-C-H aliphatic), 1689 (-COO / -
CONH stretch), 1573 (amide-II band).
1H NMR (200 MHz, DMSO-d6) b : 7.20 -7.40 (m, SH), 7.13 (d, J=8.4 Hz,
2H), 6.77 (d, J=8.6 Hz, 2H), 4.50 (t,']= 5.0 Hz, 2H), 4.29-4.22 (m, SH), 4..10
(m, 1H), 4.01 (t, J=3.0 Hz, 1H), 3.69-3.35 (m, 2H), 3.10-2.94 (m, 4H), 2.84
(t,
J=7.5 Hz, 2H), 1.90-1.78 (m, 2H), 1.40 (d, 3H), 1.16 (t; J= 7.0 Hz, 3H), 1.38
(t, J= 7.3 Hz, 3H), 0.97 (t, J= 7.4 Hz, 3H). . . _
Mass m/z : 457 (M++ 1).
Example-38
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-lI3-pyrazolo [4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid S-(+)-
phenylglycinol salt
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O Ph
N. NCO ~_~ ~ O
C-O H3N~OH
EtO O
(-)-3-[4-[2-( 1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1 H-pyrazolo[4,3-d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. 'lie reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. S-(+)-
1'henylglycinol
( 1.5 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure S-(+)-phenylglycinol salt of (-)-3-
[4-
[2-(1-methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[43-d]
pyririzidili-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white
crystalline
solid, (weighs about 5.5 g, yield : 85 %, mp : 88-90 °C, purity 99 % by
HPLC).
IR (KBr) cm 1 : 3150 (C-H aromatic), 2950 (-C-H aliphatic), 1686 (-CQO / -
CONH stretch), 1570 (amide-II band).
The 1H NMR (200 MHz, I~MS~-d6) 8 : 7.40 (m, 5H), 7.13 (d, J=8.4 Hz, 2H),
6.77 (d, J=8.6 Hz, 2H), 4.50 (t, J-- 5.0 Hz, 2H), 4.29-4.22 (m, 5H), 4.10 (in,
1H), 4.01 (t, J=3.70 Hz, 1H), 4.00 (d, 2H), 3.60-3.80 (m, 1H), 3.69-3.35 (in,
2H), 3.10-2.94 (m, 4H), 2.84 (t, J=7.5 Hz, 2H), 1.90-1.78 (in, 2H), 1.38 (t,
J=
7.3 Hz, 3H), 1.16 (t, J= 7.0 Hz, 3H), 0.97 (t, J= 7.4 Hz, 3H),
Mass m/z : 457 (M+ + 1), Anal. Calcd : C32H43N5~6~ % C 64.59; % H : 7.15;
N 11.80; Found % C 64.59; % H 7.15; % N 11.66.
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Example-39
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo [4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid aminoguanidine
hydrogen carbonate salt
O _
NH
N NCO \ _ / 101 ~ ~ ,~ 3 HCOO~
-C-O H3N N
N ~ I N Et0 ~ H
(-)-3-[4-[2-( 1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro- l I~-pyrazolo [4, 3-
d]
pyrimidiil-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Aminoguanidine
hydrogen carbonate (1.49 g) was added to the reaction mixture at 60 °O
in
about 10 minutes under stirring. Maintained gentle reflux of the reaction
mixture for 12-14 hr and monitored the progress of the reaction. The reaction
mixture was cooled to RT and stirred for 2-3 h at room temperature. The
precipitated product was filtered, Vacuum dried to afford the pure
aminoguanidine hydrogen carbonate salt of (-)-3-[4-[2-(1-Methyl-5-ethyl-7-
oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-6-yl)ethoxy]phenyl]-
2-ethoxypropionic acid as off white crystalline solid, (weighs about 5.2 g,
yield : 80 %, mp : 130 °C, purity 99% by HPLC).
IR (I~Br) cm 1 : 3450-3350 (-NH stretch), 3120 (C-H aromatic), 2970 (-C-H
aliphatic), 1676 (-COO / -CONH stretch), 1573 (amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.4 Hz, 2H), 6.77 (d, J=8.6 Hz,
2H), 4.50 (t, J= 5.0 Hz, 2H), 4.29-4.22 (m, 5H), 4.01 (t, J=3.70 Hz, 1H), 3.69
3.35.(m, 2H), 3.10-2.94 (m, 4H), 2.84 (t, J=7.5 Hz, 2H), 1.90-1.78 (m, 2H),
1.38 (tJ= 7.3 Hz, 3H), 1.16 (t, J= 7.0 Hz, 2H), 0.97 (t, J= 7.4 Hz, 3H).
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Mass m/z : 457 (M+ + 1), Anal. Calcd : C26H~oN80~; % C 54.16; % H : 6.94;
N 19.44; Found % C 54.00; % H 6.78; % N 19.32.
Example-40
5 (-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]p~rimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid tromethamine
s alt
O OH
N ~0 \ / ~ C~ OH
O O
Et0 H3~
OH
(-)-3-[4-[2-( 1-Methyl-5-ethyl-7-oxo-3-propyl-6, 7-dihydro-1 H-pyrazolo [4, 3-
d]
10 pyrimidin=6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Tromethamine
(1.32
g) was added to the reaction mixture at 60 °C in about 10 minutes under
15 stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to I~T
and stirred for 2-3 ,h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure tromethamine salt of (-)-3-[4-[2-(1-
methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-6-
20 yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white crystalline solid,
(weighs about 5.18 g, yield : 82 %, mp : 116-118 °C, purity 99 % by
HPLC).
IR (KBr) cni 1 : 3300-3250 (-OH stretch), 3120 (C-H aromatic), 2935 (-C-H
aliphatic), 1693 (-COO / -CONH stretch), 1575 (amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.4 Hz, 2H), 6.77 (d, J=8.1 Hz,
25 2H), 4.50 (t, J= 5.0 Hz, 2H), 4.29-4.22 (m, 5H), 4.01 (t, J=3.70 Hz; 1H),
4.00
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(s, 6H), 3.69-3.35 (m, 2H), 3.10-2.94 (m, 4H), 2.84 (t, J=7.5 Hz, 2H), 1.90-
1.78 (m, 2H), 1.38 .(t, J= 7.3 Hz, 3H), 1.16 (t, J= 7.0 Hz, 3H), 0.97 (t, J=
7.4
Hz, 3H). ,
Mass m/z : 457 (M+ + 1), Anal. Calcd : CZgHa3N30g; % C 61.20; % H : 7.83;
% N 7.65; Found % C 61.05; % H 7.70; % N 7.50.
Example-41
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-lA-pyrazolo [4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid
dicyclohexylamine salt
O
N NCO ~-~ O O O+ N2,
C-O
N ~ I N~ Et0
i
(-)-3-[4-[2-( 1-Methyl-5-ethyl-7-oxo-3~propyl-6,7-dihydro-1 H-pyrazolo[4,3-d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflex condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass.
Dicyclohexylamine
(1.98 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflex of the reaction mixture for 12-14 hr and
monitored the.progress of the reaction. The reaction mixture was cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure dicyclohexylamine salt of (-)-3-[4-
[2-
(1-methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-
6-yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white crystalline solid,
(weighs about 5.51 g, yield : 79 %, mp : 138-140 °C, purity 99% by
HPLC.
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82
1R (KBr) cxri 1 : 3150 (C-H aromatic), 2933 (-C-H aliphatic), 1682 (-COO / -
CONH stretch), 1571 (amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.4 Hz, 2H), 6.77 (d, J=8.6 Hz,
2H), 4.50 (t, J= 5.0 Hz, 2H), 4.29-4.22 (m, SH), 4.01 (t, J=3.70 Hz, 1H), 3.69-
3.35 (m, 2H), 3.10-2.94 (m, 4H), 2.84 (t, J=7.5 Hz, 2H), 2.60-1.00 (m, 22H),
1.90-1.78 (m, 2H), 1.38 (t, J= 7.3 Hz, 3H), 1.16 (t, J= 7.0 Hz, 3H), 0.97 (t,
J=
7.4 Hz, 3H).
Mass m/z : 457 (M+ + 1), Anal. Calcd : C36HSSNSOSi % C 67.81; % H : 8.63;
N 10.98; Found % C 67.65; % H 8.55; % N 10.75.
example-42
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro'-1H-pyrazolo [4,3-
d]pyrirriidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid N-
octylglucamine salt
O C H~-N H2-C$H~ ~
NCO ~ ~ 101 O H ' OH
N ~ I ~ rC-O . HO H
N Et0 H' OH
H ' OH
1~ CH~OH
(-)-3-[4-[2-( 1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]
pyriinidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. N-
octylglucamine
(3.2 g) was added to the reaction mixture at 60 °C in about 10 minutes
under
stirring. Maintained gentle reflux of, the reaction mia~ture for 12-14 hr and
monitored the progress of the reaction. The reaction mixture was cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
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83
filtered, vacuum dried to afford the pure, hygroscopic N=octylglucamine salt
of (-)-3-[4-[2-(1-methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white
hygroscopic solid, (weighs about 6.48 g, yield : 79 %, purity 99 % by HPLC.
IR (KBr) cm 1 : 3350 (-OH stretch), 3150 (C-H aromatic), 2929 (-C-H
aliphatic), 1689 (-COO / -CONH stretch), 1575 (amide-II band).
1H NMR (200 MHz, DMSO-d6) b : 7.13 (d, J=8.4 Hz, 2H), 6.77 (d, J=8.6 Hz,
2H), 4,50 (t, J= 5.0 Hz, 2H), 4.29-4.22 (m, 5H), 4.01 (t, J=3.70 Hz, 1H),4.20-
3.06 (m, 8H), 3.69-3.35 (m, 2H), 3.10-2.94 (m, 4H), 2.84 (t~ J=7.5 Hz, 2H),
1.90-1.78 (m, 2H), 1.40-1.00 (m, 16H), 1.38 (t, J= 7.3 Hz, 3H), 1.16 (t, J=
7.0
Hz, 3H), 0.97 (t, J= 7.4 Hz, 3H).
Mass m/z : 457 (NT~ + 1).
Example-43
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-lI3f-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid ~hT-
methylglucamine salt
.
O CH2-N H2-C H3
O ~, H OH
N ~ ~ ~ rC-O HO H
Et0 H OH
H OH
CH20H
(-)-3-[4-[2-( 1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1 H-pyrazolo [4,3-d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropioriic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. N-
methylglucamine
(2.1g) of was added to the reaction mixture at 60 °C in about 10
minutes under
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stirring. Maintained gentle reflux of the reaction mixture for 12-14 hr. and
monitored the progress of the reaction. The reaction mixture was cooled to RT
and stirred for 2-3 h at room temperature. The precipitated product was
filtered, vacuum dried to afford the pure, hygroscopic N-methylglucamine salt
of (-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxa-3-propyl-6,7-dihydro-1H-pyrazolo[4,3- '
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid as off crystalline
hygroscopic solid, (weighs about 5.7 g, yield : 80 %, purity 99 % by HPLC).
IR (KBr) cm 1 : 3400-3300 (-OH stretch), 3150 (C-H aromatic), 2935 (-C-H
aliphatic), 1676 (-COO / -CONH stretch), 1576 (amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 (d, J=8.4 Hz, 2H); 6.77 (d, J=8.6 Hz,
2H), 4.50 (t, J= 5.0 Hz, 2H), 4.29-4.22 (m, SH), 4.01 (t, J=3.70 Hz, 1H),4.00-
3.26 (m, 8H), 3.69-3.35 (m, 2H), 3.10-2.94 (m, 4H), 2.84 (t, J=7.5 Hz, 2H),
2.40 (s, 3H), 1.90-1.78 (m, 2H), 1.38 (t, J= 7.3 Hz, 3H), 1.16 (t, J= 7.0 Hz,
3H), 0.97 (t, J= 7.4 Hz, 3H).
Mass m/z : 457 (M++ 1).
Example-44
(-)-3-(4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo [4,3-
d] pyrimidin-6-yl)ethoxy]phenyl]-2~ethoxypropionic acid metformin salt
NH NH
i O
N N~~O \ / o O ~ ~ ~L .Me
--C-O . H3N N N
~ N~ Et0 H Me
(-)-3-[4-[2-( 1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1 H-pyrazolo [4, 3-
d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropariol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
heated to 55-60 °C for complete dissolution of the mass. Metformin
(1.41 g)
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as free base was added to the reaction mixture at 60 °C in about 10
minutes
under stiiTing. Maintained gentle reflux of the reaction mixture for 12-14 hr
and monitored the progress of the reaction. The reaction mixture was cooled to
RT and stirred for 2-3 h at room temperature. The precipitated product was
5 filtered, vacuum dried to afford the pure, crystalline metformin salt of (-)-
3-[4-
[2-(1-methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white
crystalline solid, (weighs about 5.45 g, yield : 85 %, rnp : 118 °C,
purity 99
by HPLC). '
10 IR (KBr) cm 1 :) 3350 (-OH stretch), 3150 (C-H arorizatic), 2950 (-C-H
aliphatic), 1677 (-COO / -CONH stretch), 1574 (amide-II band).
1H NMR (200 MHz, DMSO-d6) ~ : 7.13 (d, J=8.4 Hz, 2H), 6.77 (d, J=8.6 Hz,
2H), 4.50 (t, J= 5.0 Hz, 2H), 4.29-4.22 (m, SH), 4.01 (t, J=3.70 Hz, 1H), 3.69-
3.35 (m, 2H), 3.10-2.94 (m, 4H), 2.90 (s, 6H), 2.84 (t, J=7.5 Hz, 2H), 1.90-
15 1.78 (m, 2H), 1.38 (t, J= 7.3 Hz, 3H), 1.16 (t, J= 7.0 Hz, 3H), 0.97 (t, J=
7.4
Hz, 3H).
Mass m/z : 457 (M++ 1). Anal. Calcd : C2gH43N9Os; % C 57.43; % H : 7.35;
N 21.53; Found % C 57.29; % H 7.25; % N 21.40.
20 Example-45 '
(-)-3-[4-[2-(1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-lI3-pyrazolo [4,3-
d] pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid Lysine salt
O _ 0
NCO ~ ~ o ~ ~ H3
Et0 C O . (CH2)a.
H NH2
COOH
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(-)-3-[4-[2-( 1-Methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1 H-pyrazolo [4, 3-
d]
pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid (5.0 g), isopropanol
(50 ml) were added to 250 ml four necked round bottom flask, fitted with a
mechanical stirrer and reflux condenser. The reaction mixture was slowly
S heated to 55-60 °C for complete dissolution of the mass. Lysine
monohydrate
(1.79 g) free base was added to the reaction mixture at 60 °C in about
10
minutes under stirring. Maintained gentle reflux of the reaction mixture for
12-
14 hr and monitored the progress of the reaction. The reaction mixture was
cooled to RT and stirred for 2-3 h at room temperature. The precipitated
product was filtered, vacuum- dried to afford the pure lysine salt of (-)-3-[4-
[2-(1-methyl-5-ethyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-
d]pyrimidin-6-yl)ethoxy]phenyl]-2-ethoxypropionic acid as off white
amorphous solid, weighs about 5.77 g, yield : 85 %, mp : 162-164 °C,
purity
99 % by HPLC).
IR (KBr) em 1 : 3120 (C-H aromatic), 2965 (-C-H aliphatic), 1691 (-COO l
-CONH stretch), 1573 (amide-II band).
1H NMR (200 MHz, DMSO-d6) 8 : 7.13 ,(d, J=8.4 Hz, 2H), 6.77 (d, J=8.6 Hz,
2H), 4.50 (t, J= 5.4 Hz, 2H), 4.29-4.22 (m, SH), 4.20 (t, 1H), 4.01 (t, J=3.70
Hz, 1H), 3.69-3.35 (m, 2H), 3.10 (t, 2H), 3.10-2.94 (m, 4H), 2.90 (s, 6H),
2.84
(t, J=7.5 Hz, 2H), 2.11-1.5 (m, 6H), 1.90-1.78 (m, 2H), 1.38 (t, J= 7.3 Hz,
3H),
1.16 (t, J= 7.0 Hz, 3H), 0.97 (t, J= 7.4'Hz, 3H).
Mass m/z ; 457 (M+ + 1). Anal. Calcd : C3oH48N6O8; % C 58.06; % I~ : 7.74;
N 13.54; Found % C 57.9; % H 7.55; % N 13.38.
(-) .2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoXy]phenyl]propanoic acid was prepared according to the procedure
described in our copending application No. 09/507,371 :
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A solution of [2S, N(1S)]-2-ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-
dihydropyrimidin-1-yl]ethoxy]phenyl]-N-(2-hydroxy-1-phenylethyl)
propanamide (295 mg, 0.53 inmol) in a mixture of 1M sulfuric acid. (7.7 mL)
and dioxane / water (1 : 1, 14 mL) was heated at 90 °C for 48 h and the
phi of
the mixture was adjusted to 4 by the addition of aqueous sodium bicarbonate
solution. The mixture was extracted with ethyl acetate and the combined ethyl
acetate layers were washed with water, brine, dried (NazS04) and evaporated
to yield (-) 2-ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid as a colorless solid.
~Iowever, any other procedure for preparing (-) 2-ethoxy-3-[4-[2-[2-ethyl-6-
oxo-4-phenyl-1,6-dihydropyrimidin-1-yl]ethoxy]phenyl]propanoic acid can be
used. (~) 2-ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid and (+) 2-ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-
phenyl-1,6-dihydropyrimidin-1-yl]ethoxy]phenyl]propanoic acid can be
prepared by a similar procedure described above. .
Example 46
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid potassium salt
O
Ph N~ j , O
N\/\ \ I H OC2H5
O
A mixture of (-) 2-ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-
dihydropyrimidin-1-yl]ethoxy]phenyl]propanoic acid (5.0 g) and isopropanol
(75 ml) was refluxed to obtain clear solution. Aqueous potassium hydroxide
solution (0.63 g in 2 ml water) was added slowly at refluX temperature under
stirring and continued stirring for 2-3 h. Then the reaction mass was
cooled~to
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room temperature and continued stirring for further 12 h. The precipitated
solid. was filtered and dried to yield the title compound, (weights about 4.5
g,
mp (DSC) 240.9 °C).
1R (KBr) cni 1 . 3439.8, 2976.2, 2934.3, 2877:4, 1667.1, 1602.7, 1551.4,
1509.9, 1448.7, 1407.3, 1363.2, 1241.8,1178.2, 1108.7, 957.9, 898.3, 864.2,
815.7, 781.9, 696.6, 643.6.
p- XRD: 5.56, 7.46, 8.30, 14.36, 14.72, 15.74, 16.84, 17.06, 17.88,
18.40, 18.68, 19.44, 24.30, 24.42, 24.92 (2A).
Exariiple 47
(-) 2-Ethoxy-3-(4-[2-[2-ethyl-6-oxo-4-phenyl-1,6=dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid sodium salt
O
Ph [ N~ , I '. O
O ~" H OC2H5
O
1~ mixture of (-) 2-ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-
dihydropyrimidin-1-ylJethoxy]phenylJpropanoic acid (5.0 g) and isopropanol
(75 ml) was refluxed to obtain clear solution. Sodium methoXide (0.615g) v~ias
added at reflux temperature under stirring and continued stirring for 5-6 h.
Then the reaction mass was cooled to room temperature, filtered and dried to
yield the title compound (weights about 4.6 g, mp (DSC) 251.19 °C).
IR (KBr) cm 1 : 3437.2, 2977.7, 2879.1, 1667.7, 1605.4, 1552:5, 1509.7,
1449.0, 1407.6, 1362.9, 1282.8, 1242.0, 1178.2, 1108.9, 1047.4, 958.6, 898.9,
863.4, 815.4, 781.3, 737.5, 696.1, 644.,6, 519Ø
p- XRD: 5.52, 7.38, 8.24, 14.42, 14.68, 17.0, 17.88, 19.46, 19.80, 23.38,
24.26, 24.44, 26.40 (2~).
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Example 48
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid L-arginine salt
O NH2
P h I N~~ / I , O ~ H N N ~~
N~ ~ H OC2H5 ~ H C02H
O O+ NH3
A , mixture of (-) 2-ethoxy-3-r4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-
dihydropyrimidin-1-yl]ethoxy]phenyl]propanoic acid (10 g) and isopropanol
(200 ml) was refluxed to obtain clear solution. Aqueous L arginine solution
(3.99 g in 6 ml water) was added slowly at reflux temperature under stirring
and continued stirring for 2-3 h. Then the reaction mass was cooled to room
temperature and continued stirring for further 12 h. The precipitated solid
was
filtered and dried to yield the title compound (weights about 8.5 g, mp (DSC)
187 °G).
IR (hBr) cni 1 : 3358.9, 3064.4, 2974.3, 1665.5, 1591.0, 1544.9, 1511.2,
1448.9, 1406.3, 1323.7, 1240.1, 1178.9, 1111.2, 1046.2, 956.8, 896.7, 780.5,
697.3, 542:0.
p- XRD: 3.68, 7.34, 11.34, 12.74, 14.18, 14.86, 15.12, 16.64, 19.48, 20.0,
20.54, 20.86, 22.70, 30.02 (20).
Example 49
(-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-g,fi-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid L-lysine salt
O
NH3
Ph I N~ / [ ' O0 (CH2)4
N~ ~ H OC2H5 H-~--NH2
O
O COOH
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A mixture of (-) 2-ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-
dihydropyrimidin-1-yl]ethoxy]phenyl]propanoic acid (5.0 g) and isopropanol
(75 ml) was refluxed to obtain clear solution. Aqueous L lysine solution (1.88
g in 3m1 water) was added slowly at reflux temperature under stirring and
5 continued ~ stirring for 12-15 h. Then the reaction mass was cooled to room
temperature and the precipitated solid was filtered and dried to yield the
title
compound (weights about 4.3 g, mp (D8C) 114.93 °C).
IR (KBr) cni l : 3413.9, 2934.8, 1663.6, 1571.6, 15457, 1511.8, 1449.0,
1405.5, 1323.2, 1241.8, 1179.3, 1112.1, 1046.5, 956.0,898.9, 853.9, 782.5,
10 736.6, 698.9, 644.7, 557.8.
p- ~: 4.42, 7.58, 7.84, 7.98, 11.64, 11.96, 13.62, 15.76, 16.14, 16.36,
16.54, 17.84, 19.32, 19.46 (20).
Example 50
15 (-) 2-Ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-dihydropyrimidin-1-
yl]ethoxy]phenyl]propanoic acid t-butyl amine salt
O
Ph I N~ ~ I ~ ~ NH3
-IN ~ H OC2H5 CH3--~-CH3
CHg
O
A ' mixture of (-) 2-ethoxy-3-[4-[2-[2-ethyl-6-oxo-4-phenyl-1,6-
dihydropyrimidin-1-yl]ethoxy]phenyl]propanoic acid (2.0 g) and isopropanol.
20 (15 ml) was heated to reflux to get the clear solution. t-Butylamine (0.35
g)
was added slowly dropwise and continued reflux for 2-3 hr. Then the reaction
mass was cooled to room temperature and continued stirring for further 2 h.
Then slowly reaction mass cooled to 15 °C, continued stirring for
overnight.
Filtered the compound at 10-15 °C, washed with isopropanol (5 ml)
dried
25 under high vacuum at 50-60 °C over a period of 8-10 hr to yield the
title
compound (weights 'about 2.0 g).
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DSC: 90.89-105.46 (Endo), 118.30 (Exo), 158.37 (Endo)
p-XRD: 4.98, 6.84, 8.52, 10.14, 13.34, 14.64, 15.76, 17.32, 18.16, 19.54,
20.62, 22.20, 23.10, 24.76, 25.60, 27.36, 31.54, 32.46 Cm 1.
IR: 3428, 2977, 1676, 1545, 1512, 1470, 1448, 1402, 1319 1238, 1113, 1047,
900, 854, 781, 699Cm 1.
The compounds of the present invention lowered random blood sugar
level, triglyceride, total cholesterol, LDL, VLDL and increased HDL. This
was demonstrated by ih vitro as well as ih vivo animal experiments.
Demonstration of Efficacy of Compounds
A) Iya vaty~~
a) Determination of hPPARa, activity
Ligaiid binding domain of hPPARoc was fused to DNA binding domain
of Yeast transcription factor GAL4 in eucaryotic expression vector. Using
superfect (Qiagen, Germany) as transfecting reagent HEK-293 cells were
transfected with this plasmid and a~reporter plasmid harboring the luciferase
a gene driven by a GAL4 specific promoter. Compound was added at different
concentrations after 42 hrs of transfection and incubated overnight.
Luciferase
activity as. a function of compound binding/activation capacity of PPARoc was
measured using Packard Luclite kit (Packard, USA) in Top Count (Ivan
Sadowski, Brendan Bell, Peter Broag and Melvyn Hollis. Gene. 1992. 118
137 -141; Superfect Transfection Reagent Handbook. February 1997. Qiagen,
Germany).
b) Determination of hPPARy activity
Ligand binding domain of hPP'ARy1 was fused to DNA binding domain
of Yeast transcription factor GAL4 in eucaryotic expression vector. Using
lipofectamine (Gibco BRL, USA) as transfecting reagent HEK-293 cells were
transfected with this plasmid and a reporter plasmid harboring the luciferase
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92
gene driven by a GAL4 specific promoter. Compound was added at 1 ~M
concentration after 48 hrs of transfection and incubated overnight. Luciferase
activity as a function of drug binding/activation capacity of PPARyl was
measured using Packard Luclite kit (Packard, ~JSA) in Packard Top Count
(Ivan Sadowski, Brendan Bell, Peter Broag and Melvyn Iiollis. Gene. 1992.
118 .: 137 -141; Guide to Eukaryotic Transfections with Cationic Lipid
Reagents. Life Technologies, GIBCO BRL, IJSA).
~xannple ConcentrationppARoc PPARy Concentration
No.
11 ~ 50 ~M 13.8 1 p,M 27.0
16 S 0 ~M 3.4 1 ~M 13.5
20 50 ~M 3.5 1 ~M 13.0
23 50 ~M 3.4 1 ~,M 11.5
36 50 pM 3.3 1 p,M 11.4
38 ' SOp.M 3.3e l,uM 12.0
41 50 p.M 3.1 1 ~,M 13.0
44 50 ~,M 2.5 1 ~M 12.0
c) Determination of I~1VIG CmE1 reductase inhibition activity
Liver microsome bound reductase is prepared from 2% cholestyramine
fed rats at mid-dark cycle. Spectrophotometric assays are carried out in 100
mM I~HaP04, 4 mM DTT, 0.2 mM NADPH, 0.3 mM HMG CoA and 125 ~.g
of liver microsomal enzyme. Total reaction mixture voluriie is kept as 1 ml.
Reaction is started by addition of HMG CoA. Reaction mixture is incubated at
37°C for 30 min and decrease in absorbance at 340 nm is recorded.
Reaction
mixture without substrate is used as blank (Goldstein, J. L and Brown, M. S.
Progress in understanding the LDL receptor and HMG CoA reductase, two
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93
membrane, proteins that regulate the plasma cholesterol. J. Lipid Res. 1984,
25: 1450 - 1461). The test compounds will inhibit the HMCp CoA reductase
enzyme.
'In vavo
a) Efficacy in genetic modeYs
Mutation in colonies of laboratory animals and different sensitivities to
dietary regimens have made the development of animal models with non-
insulin dependent diabetes and hyperlipidemia associated with obesity and
insulin resistance possible. Genetic models such as db/db and ob/ob
(Diabetes, (1982) 31(1) : 1- 6) mice and zucker fa/fa rats have been developed
by the various laboratories for understanding the pathophysiology of disease
and testing the efficacy of new antidiabetic compounds (Diabetes, (1983) 32:
830-838 ; Annu. Rep. Sankyo Res. Lab. (1994). 46 : I-57). 'The homozygous
animals, C57 BL/KsJ-db/db mice developed by Jackson Laboratory, I_TS, are
obese, hyperglycemic, hyperinsulinemic and insulin resistant (J. Clin.
Invest.,
(1990) 85 : 962-9'67), whereas heterozygous are lean and norinoglycemic. In
db/db model, mouse progressively develops insulinopenia with age, a feature
commonly observed in late stages of human type II diabetes when blood sugar
levels are insufficiently controlled. The state of pancreas and its course
vary
according to the models. Since this model resembles that of type II diabetes
mellitus, the compounds of the present invention were tested for blood sugar
and triglycerides lowering activities.
Male C57BL/KsJ-db/db mice of 8 to I4 weeks age, having body weight
range of 35 to 60 grams, bred at Dr. Reddy's Research Foundation (DRF)
animal house, were used in the experiment. The mice were provided with
standard feed (National Institute of Nutrition (NIN), Iiyderabad, India) and
acidified water, ad libitum. 'The anirilals having more than 350 mg / dl blood
sugar were used for testing. The number of animals in each group was 4.
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Test compounds were suspended on 0.25 % carboxymethyl cellulose
and administered to test group at a dose of 0.1 mg to 30 mg / kg through oral
gavage daily for 6 days. The control group received vehicle (dose 10 ml / kg).
On 6th day the blood samples were collected one hour after administration of
test compounds / vehicle for assessing the biological activity.
The random blood sugar and triglyceride levels were measured ~ by
collecting blood (100 ~1) through orbital sinus, using heparinised capillary
in
tubes containing EDTA which was centrifuged to obtain plasma. The plasma
glucose and triglyceride levels were measured spectrometrically, by glucose
oxidase and glycerol-3-P04 oxidase/peroxidase enzyme (I~r. Reddy's Lab.
I3iagnostic Division Kits, Hyderabad, India) methods respectively.
The blood sugar and triglycerides lowering activities of the test
compound was calculated according to the formula.
No adverse effects were observed for any of the mentioned compounds
of invention in the above test.
example Dose (mg / Reduction in Blood Glucose
No. kg) )Level (/~)
Fxample 0.1 61
11
The ob/ob mice are obtained at 5 weeks of age from Bomholtgard,
Denmark and are used at 8 weeks of age. Zucker falfa fatty rats are obtained
from IffaCredo, France at 10 weeks of age and are used at 13 weeks of age.
The animals are maintained under 12 hour light and dark cycle at 25 + 1
°C.
Animals are given standard laboratory chow (N1N, Hyderabad, India) and
water, ad libitum (Fujiwara, T., Yoshioka, S., Yoshioka, T., ZJshiyama, I and
Horikoshi, H. Characterization of new oral antidiabetic agent CS-045. Studies
in KK and ob/ob mice and Zucker fatty rats. Diabetes. 1988. 37 : 1549 -
1558).
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The test compounds will be administered at 0.1 to 30 mg/kg/day dose
for 9 days. The control animals receives the vehicle (0.25%
carboxymethylcellulose, dose 10 ml/kg) through oral gavage.
The blood samples can be collected in fed state 1 hour after drug
5 administration on 0 and 9 day of treatment. The blood can be collected from
the retro-orbital sinus through heparinised capillary in EDTA containing
tubes.
After centrifugation, plasma sample will be separated for triglyceride,
glucose,
free fatty acid, total cholesterol and insulin estimations. Measurement of
plasma triglyceride, glucose, total cholesterol can be done using commercial
10 kits (Dr. Reddy's Laboratory, Diagnostic Division, India). The plasma free
fatty acid will be measured using a commercial kit from Boehringer
Mannheim, Germany. The plasma insulin can be measured using a RIA kit
(BARC, India). The reduction of various parameters examined will be
calculated according to the formula given below.
15 In ob/ob mice oral glucose tolerance test is performed after 9 days
treatment. Mice are fasted for 5 hrs and challenged with 3 ,gm/kg of glucose
orally. The blood samples are collected at 0, 15, 30, 60 and 120 min for
estimation of plasma glucose levels.
b) Plasma tri~lyceride and Cholesterol lowerin~activity in
20 hypercholesterolemic rat models
Male Sprague Dawley rats (NIN stock) are bred in DRF animal house.
Animals are maintaW ed under 12 hour light and dark cycle at 25 ~
1°C. Rats
of 180 - 200 gram body weight range were used for the ea~periment. Animals
are made hypercholesterolemic by feeding 2% cholesterol and 1 % sodium
25 cholate mixed with standard laboratory chow [National Institute of
Nutrition
(NIN), Hyderabad, India] for 6 days. Throughout the experimental period ,the
animals are maintained on the same diet (Petit, D., Bonnefis, M. T., Rey, C
and Infante, R. Effects of ciprofibrate on liver lipids and lipoprotein
synthesis
a
in norino- and hyperlipidemic rats. Atherosclerosis. 1988. 74 : 21S - 225).
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The test compounds can be administered orally at a dose 0.1 to 30
mg/kg/day for 3 days. Control group is treated with vehicle alone (0.25
Carboxymethylcellulose; dose 10 ml/kg).
The blood samples can be collected in fed state 1 hour after drug
administration on 0 and 3 day of compound treatment: The blood can be
collected from the retro-orbital sinus through heparinised capillary in EDTA
containing tubes. After centrifugation, plasma sample will be separated for
total cholesterol, HDL and triglyceride estimations. Measurement of plasma
triglyceride, total cholesterol and HDL are done using cbmmercial kits (Dr.
Reddy's Laboratory, Diagnostic Division, India). LDL and VLI~L cholesterol
can be calculated from the data obtained for total cholesterol, HDL and
triglyceride. The reduction of various parameters examined are calculated
according to the formula given below.
c) Plasma tri~lyceride and total cholesterol lowering activity in
Swiss albino mice and Guinea pigs
Male Swiss albino mice (SAM) and male Guinea pigs were obtained
from NAT and housed in DRF animal house. All these animals were
maintained under 12 hour light and dark cycle at 25 ~ 1 °C. Animals
were
given standard laboratory chow (NIN, Hyderabad, India) and water, ad
libitum. SAM of 20 - 25 g body weight range and Guinea pigs of 500 - 700 g
body weight range were used (Oliver, P., Plancke, M. O., Marzin, D., Clayey,
V., Sauzieres, J and Fruchart, J. C. Effects of fenofibra~e, gemfibrozil and
nicotinic acid on plasma lipoprotein levels in normal and hyperlipidemic mice.
Atherosclerosis. 1988. 70 : 107 -114).
.The test compounds were administered orally to Swiss albino mice at
0.3 to 30 mg/kg/day dose for 6 days. Control mice were treated with vehicle
(0.25% Carboxymethylcellulose; dose 10 ml/kg). The test compounds were
administered orally to Guinea pigs at 0.3 to 30 mg/kg/day dose for 6 days.
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Control animals were treated with vehicle (0.25% Carboxymethylcellulose;
dose 5 ml/kg).
The blood samples were collected in fed state 1 hour after drug
administration on 0 and 6 day of treatment. The blood was collected from the
retro-orbital sinus through heparinised capillary in ED'I'A containing tubes.
After centrifugation, plasma sample was separated for triglyceride and total
cholesterol (Wieland; ~. Methods of Enzymatic analysis. Bergermeyer, H. O.,
Ed., 1963. 211 - 214; Trinder, P. Ann. Olin. Biochem. 1969. 6 : 24 - 27).
Measurement of plasma triglyceride, total cholesterol, and HDL were done
using commercial kits (Dr. Reddy's Diagnostic Division, Hyderabad, India).
Example Dose (mg / kg) Triglyceride Lowering
No. (%)
1 3 ' 87
3 3 81
11 0.3 58
12 0.3 37
31 3 84
44 3 80
45 3 81
d) Body weight reducing effect in cholesterol fed hamsters
Male Syrian Hamsters are procured from NIN, Hyderabad, India.
Animals are housed at DRF animal house under 12 hour light and dark cycle
at 25 ~ 1°C with free access to food and water. Animals are maintained
with
1 % cholesterol containing standard laboratory chow (N1N) from the day of
treatment.
The test compounds can be administered orally at 1 to 30 mg/kg/d~ay
dose for 15 days. Control group animals are treated with vehicle (Mill Q
water, dose 10 ml/kg/day). Body weights are measured on every 3rd day.
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Formulae for calculation
1. Percent reduction in Blood sugar / triglycerides / total cholesterol were
calculated according to the formula
TT / OT
Percent reduction (%) = 1 - X 100
TC / OC
OC = Zero day control group value
OT = Zero day treated group value
TC = Test day control group value
TT = Test day treated group value
2. LDL and VLDL cholesterol levels were calculated according to the
formula
Triglyceride
LDL cholesterol in mg/dl = - [ Total cholesterol - HDL cholesterol - ] mg/dl
5
VLDL cholesterol in mg/dl = [Total . cholesterol - HDL cholesterol - LDL
cholesterol] mg/dl.
Single dose oral pharmacokinetic studies
Male VVistar rats (220 - 250 gm) were used in the experiments. The
animals were maintained under standard laboratory conditions and had free
access to feed and water ad libitum. Before experimentation animals were
fasted overnight (~15 h) during which they had free access to water ad
libitum.
An amount equivalent to 30 mg of drug was weighed accurately and
transferred into a clean mortar and triturated to obtain a fine powder. To
this
0.5 ml of 0.25% sodium carboxy methyl cellulose (sodium CMC) was added
to obtain a paste. To the obtained paste remaining 2.5 ml of sodium CMC was
added to make up the volume to 3 ml. Based on the animal weight appropriate
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volume (body weight x 3) of the prepared suspension was administered
through oral gavage.
After dosing, at designated time points (0.5, 1, 2, 3, 5, 8, 12 and 24 h)
200 ~Zl of blood was collected from retro orbital plexus into 0.5 ml
eppendorff
tubes containing EDTA (10 g.1 of 200 mg/ml solution in Milli Q water). Blood
was centrifuged at 12,800 rpm for 5 min and obtained plasma and stored at -
20 °C till further analysis.
1001 plasma was transferred into a clean and dry centrifuge tube. To
this internal standard (10 ~l of 100 ~g/ml) was added and extracted With 2 ml
of extraction recovery solvent. The contents were vortexed for 2 min, followed
by centrifugation for 10 min at 2800 rpm. Clear organic layer (2 x 0.75 ml)
was separated and dried under nitrogen gas at 50 °C: The residue was
reconstituted with 150 w1 of mobile phase and vortexed for 20 sec, from this
50 ~.1 was injected onto HPLC column.
Pharmacokinetic parameters were calculated by non-compartmental
model analysis. The peak plasma concentration (CmaX) and the corresponding
time (TmaX) were directly obtained from the raw data. The area under the
plasma concentration versus time curve up to the last quantifiable time point,
AUC~o_t~ was obtained by the linear and log-linear trapezoidal summation. The
AUC~o_t~ extrapolated to infinity (i.e., AUC ~0_~1) by adding the quotient of
Clast~ela Where Coast represents the last measurable time concentration and
I~el
represents the apparent terminal rate constant. I~~ was calculated by the
linear
regression of the log-transformed concentrations of the drug in the terminal
phase. The half life of the terminal elimination phase was obtained using the
relationship tyz = 0.693/ I~~.
Example AUC ~o_~) AUC ~o_t~Cn,ax Tn,ax W (h') $n2
(h) (h)
No. (pg.hrlml (~g.hr/ml(~,g/ml)
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4 38.36 6.7640.55 38.67 0.50 0.31 1.97
+ +
6.93 10.76 0.00 0.15 0.46
7 51.43 16.2451.65 40.25 0.90 0.38 1.91
+ +
16.34 16.93 0.22 0.09 0.41
74.07 21.2675.89 54.21 0.50 0.34 2.16
21.03 19.66 0.00 0.08 0.58
31 65.93 15.5464.70 19.15 2.75 0.34 2.24
+
15.82 5.01 1.50 Ø12 0.87
40 78.55 19.4977.80 34.19 0.75 0.60 1.20
19.27 10.03 0.29 0.13 0.25
~
45 , 101.29 99.33 24.22 2.38 0.42 1.72
14.76 15.42 9.94 2.06 0.09 0.38
.
47 83.28 10.4582.25 39.54 0.50 0.34 2.07
+
10.61 10.43 0.00 0.04 0.22
46 62.91 8.7861.25 17.69 1.33 0.32 2.19
+
8.86 5.74 ' 0.75 0.03 0.20
48 92.57 27.28'90.12 50.44 0.50 0.29 2.18
28.59 11.12 0.00 0.11 0.61
