DERIVATIVES OF PHENYLALKYL AND PHENOXYALKYL ACIDS FOR THE TREATMENT OF THE HYPERGLYCAEMIA, HYPERTRIGLYCERIDAEMIA AND TYPE 2 DIABETES AND PROCESS FOR PREPARING THEM
The invention described herein relates to the preparation of new derivatives of mono- and dicarboxylic phenyl or phenoxyal yl acids useful for the treatment of the hyperglycaemia and hypertxiglyceridaemia typical of type 2 diabetes. Background to the invention Diabetes is a widespread disease throughout the world and is associated with major clinical complications including macrovascular (atherosclerosis) and microvascular (retinopathy, nephropathy and neuropathy) damage. Such complications are inevitable consequences of the disease and constitute a serious threat to the subject's life and well-being. Diabetes is associated with various abnormalities such as obesity, hypertension and hyperlipidaemia. Various clinical forms of diabetic disease are known, the most common being type 2 and type 1 diabetes. Type 2 diabetes is characterised by reduced sensitivity to the action of insulin (insulin resistance) and gives rise to an increase in actual insulin levels in the body in an attempt to compensate for this deficiency and to a consequent increase in glucose levels. Numerous reports have confirmed the involvement of insulin resistance in many disease conditions in addition to type 2 diabetes itself, such as dyslipidaemia, obesity, and arterial hypertension. The combination of insulin resistance and obesity, hypertension and dyslipidaemia is known as Syndrome X.
Drugs used for many years such as the biguanides and sulphonylurea drugs are available on the market for the treatment of type 2 diabetes. In the case of the biguanides (the best known of which is metformin) the mechanism of action is still unclear; it presents side effects such as acidosis and gastrointestinal disorders and is contraindicated in renal, cardiac and pulmonary insufficiency. Sulphonylurea drugs promote the secretion of insulin by the β -cells and may present episodes of hypoglycaemia as a possible side effect. Drugs recently introduced onto the market are the thiazolidinediones, i.e. insulin- sensitising antidiabetic compounds such as troglitazone (J. Med. Chem., 1989, 32, 421-428), pioglitazone (Arzneim. Forsch./ Drug Res., 1990,, 40 (1), 37-42), and rosiglitazone (Bioorg. Med. Chem. Lett, 1994, 4, 1181- 1184) which are capable of reducing hyperglycaemia and insulin levels. These compounds are high-affinity synthetic ligands of PPARy (J. Biol. Chem., 1995, 270, 12953- 12956).
Peroxisome proliferator activated receptors (PPARs) are receptors belonging to the superfamily of nuclear receptors whose function is to control the expression of genes involved in carbohydrate and lipid metabolism (J. Med. Chem., 2000, 43, 527- 550). Various subtypes of PPARs have been identified: PPARγ, PPARα and PPARβ (also known as PPAR δ). The gamma isoform (PPARy) is involved in the regulation of the differentiation of adipocytes and in
energy homeostasis, whereas the alpha isoform (PPARα) controls fatty acid oxidation. In structure -activity relationship studies aimed at identifying new molecules endowed with potential antidiabetic action, a correspondence has been confirmed between PPARy activation and serum glucose lowering activity (J. Med. Chem., 1996, 39, 665-668; J. Med. Chem., 1998, 41, 5020-5036; 5037-5054; 5055-5069). The insulin- sensitising action would appear to be related, as far as this first series of compounds is concerned, to the fatty acid recruitment action regulated by activated PPARy which is thought to lead to an improvement in the insulin resistance of the tissues, enhancing serum glucose levels and lowering insulin levels. (Diabetes, 1998, 47, 507-514).
The side effects already observed with troglitazone and feared also in the case of the other compounds of this class are: severe liver toxicity (which caused the withdrawal of troglitazione from the US market), increased cholesterol, weight gain and oedema. In recent years molecules with a mixed profile, i.e. ligands of PPARy and PPARα have emerged (KRP 297, Diabetes, 1998, 47, 1841- 1847; DRF 2725, Diabetes, 2001, 50, suppl. 2, A108; AZ 242, Diabetes, 2001, 50, suppl. 2, A121-A122). These compounds are potentially capable of exerting a good measure of control of diabetic disease, while presenting a serum glucose and serum lipid lowering action with fewer side effects typical of the first series of compounds in the thiazolidinedione class, consisting exclusively of PPARy ligands. Not
all the scientific community, however, agrees with this line of thinking. Recent studies on new-generation compounds, whether thiazolidinedione derivatives or otherwise (MC555, J. Biol. Chem., 1998, Vol. 273 (49), 32679-32684; NC2100 Diabetes, 2000, 49, 759- 767, YM440, Metabolism, 2000, 49, 41 1-417), in gene transactivation tests, in-vitro glucose uptake experiments with muscle tissue and in-vivo experiments in transgenic animals with deficient PPARy expression, have led to the hypothesis that there is no real direct relationship between PPARy activation and the serum glucose and serum lipid lowering activity of these compounds (Toxicology Letters, 2001, 120, 9- 19). In particular, for YM440, a serum glucose lowering activity has been reported which is not attributable to PPARy activation, in that adipocyte differentiation and the consequent increase in body weight due to the increased fat mass are not observed. This strengthens the hypothesis that the serum glucose lowering activity of these molecules is not necessarily related to PPARy activation and that these compounds may be capable of modulating carbohydrate and lipid metabolism through interaction with other biochemical targets. This is confirmed by the work of investigators who have opted for the use of in-vivo screening in diabetic animals (db/db mice, ob/ob mice) in order to identify possible insulin- sensitising agents which are not necessarily good PPAR ligands. These experiments have led to the selection of compounds still being investigated with promising antidiabetic
activity in animal models (DRF 2189, J. Med. Chem., 1998, 41, 1619- 1630; JTT-501, J. Med. Chem., 1998, 41, 1927- 1933).
In conclusion, then, since the first compounds belonging to the thiazolidinedione class (troglitazone in particular) have proved to be associated with substantial hepatotoxic and other side effects, probably related to their PPARy activity, the scientific community would now appear to be oriented towards the search for new compounds with a different mechanism of action which induce a similar or better effect on insulin sensitivity and glucose homeostasis without toxic side effects (J. Med. Chem., 2001, 44, 2601-2611).
We should also recall that, in addition to the above-mentioned YM440, the compounds of patents WO 93/03021 (Yamanouchi), WO 01 /79150 Al (Novo Nordisk), and US 5,063,240 (Beecham Group) have structures analogous to those of the compounds according to the present invention; in these patents, however, only in-vitro assays are reported, and only in one case (Beecham Group) is the ability to lower serum glucose demonstrated in db/db and ob/ob mice. Summary of the invention It has now been found that compounds with formula (I) have been reported as being active as serum glucose and serum lipid lowering agents and are endowed with low toxicity and are therefore useful as medicines, particularly for the treatment of hyperglycaemias and hyperlipidaemias.
The preferred applications are the prophylaxis and treatment of diabetes, particularly type 2 diabetes and its complications, Syndrome X, the various forms of insulin resistance and hyperlipidaemias .
The object of the invention described herein are formula (I) compounds:
in which: m = 0, 1; n = 0 - 4; when n = 0, m = 0 and the two aromatic rings are bound to form a biphenyl group; when n is from 2 to 4 the alkyl chain can be saturated or unsaturated, R3 and R4 can be the same or different and can be selected from H and alkyl C1-C5;
Zi and Z2 = 0, 1 and can be the same or different; Y can be O, -CH= or -CH2 when Zi and Z2 are equal to 1 , or OH when the corresponding Zi or Z2 is equal to zero; X can be -OH, -O-alkyl C1-C3;
Rl and R2 can be the same or different, and can be selected from the group consisting of: -H; alkyl C1 -C5; alkoxy possibly substituted with halogens; phenoxy possibly substituted with halogens, nitro, hydroxy, alkyls; benzyloxy possibly substituted with halogens, nitro, hydroxy, alkyls; COX; their pharmacologically acceptable salts, racemic mixtures, the single enantioners, geometric isomers or stereoisomers and tautomers, with the proviso that formula I compounds are excluded in which:
R3 = R4 = CH3; m = 0, n = 1; Y = 0;
Rl = R2 = CH3; X = OH, -O-alkyl C1-C3;
Zi = Z2 = 1; because these are described in patent DE 2017331 (1970). A further object of the present invention is the use of formula I compounds as medicines. A further object of the present invention is the use of formula
I compounds for the preparation of a medicine for the treatment of hyperglycaemias and hyperlipidaemias, particularly for the treatment of type 2 diabetes and its complications.
Further objects of the present invention are pharmaceutical compositions containing as their active ingredient a formula I compound and at least one pharmaceutically acceptable excipient and /or diluent. These and other objects will now be described in detail also by means of examples which illustrate but do no not limit the invention.
Detailed description of the invention
In the formula (I) compounds, a first group of preferred compounds consists of compounds in which m = 1 , n is from 1 to 4, preferably 2 or 4, and the alkyl can be saturated or unsaturated, R3 and R4 are the same and preferably equal to H, Y can be O, -CH= or
-CH2 and Zi may or may not be the same as Z2; if Zi is not the same as Z2 the latter is preferably equal to zero and the corresponding Y is OH. Within the context of this first group, Rl can be the same as R2 and equal to hydrogen or alkyl, preferably methyl; or Rl is preferably
COX, with X equal to -O-alkyl, preferably methyl, and R2 is H.
A second group of preferred compounds consists of compounds in which m = 0 and n = 0, or m = 0 and n is preferably from 1 to 2, and in this case the alkyl can be saturated or unsaturated, preferably saturated, R3 and R4 are the same and can be equal to H or to alkyl, preferably methyl, Y is preferably O and Zi may or may not be the same as Z2; if Zi is not the same as Z2, the latter is preferably equal to zero and the corresponding Y is OH. In
the context of this second group, Rl may or may not be the same as R2; if Rl is the same as R2, it is preferably alkyl, preferably methyl; if Rl is different from R2, Rl is preferably COX, with X equal to -O- alkyl, preferably methyl, and R2 is preferably H. A third group of preferred compounds consists of compounds where m and n are equal to zero, Y is preferably -CH=, -CH2-, and Zi is the same as Z2. In the context of this group, Rl is preferably COX, with X equal to -O-alkyl, preferably methyl, and R2 is H. Even more preferred are the following compounds: 1. Dimethyl 2-[4-(2~{4-[3-methoxy-2-(methoxycarbonyι)-3- oxopropyl]-phenoxy}ethoxy)benzyl]malonate (ST1720); 2. Dimethyl 2-{4'-[3-methoxy-2-(methoxycarbonyl)-3-oxo- l- propenyl]- [ 1 , l'-biphenyl]-4-yl}- 1 , 1 -ethylenedicarboxylate (ST2013); 3. Dimethyl 2-({4'-[3-methoxy-2-(methoxycarbonyl)-3- oxopropyl] [1, 1 '-biρhenyl]-4-yl}methyl)malonate (ST2032) ;
4. Dimethyl 2-{4-[((2Z)-4-{4-[3-methoxy-2-(methoxycarbonyl)-3- oxo- 1 -propenyl]phenoxy}-2-butenyT)oxy]phenyl}- 1 , 1- ethylenedicarboxylate (ST2012);
5. Dimethyl 2-{4-[((2Z)-4-{4-[3-methoxy-2-(methoxycarbonyl)-3- oxo-proρyl]phenoxy}-2-butenyl)oxy]benzyl}malonate (ST2145);
6. Dimethyl 2-[4-(4-{4-[3-methoxy-2-(methoxycarbonyl)-3- oxopropyl] -phenoxy}butoxy)benzyl]malonate (ST2144);
7. Methyl 2-{3-[2-(3-hydroxyphenoxy)ethoxy]phenoxy}-2-methyl- propanoate (ST 1877);
8. Methyl 2-(3-{2-[3-(2-methoxy- l, l-dimethyl-2-oxoethoxy)- phenoxy]ethoxy}phenoxy) -2-methylpropanoate (ST 1878) ;
9. Dimethyl 2-{4-[l-(4-hydroxyphenyl)- l-methylethyl]-phenoxy}- malonate (ST2020)
10. Dimethyl 2-[4-(l-{4-[2-methoxy- l-(methoxycarbonyl)-2- oxoethoxy]phenyl}- 1 -methylethyl)phenoxy]malonate (ST2048) ;
1 1. Methyl 2-{[4'-(2-methoxy- l, l-dimethyl-2-oxoethoxy)[l , l'- biphenyl]-4-yl]oxy}-2-methylpropanoate (ST2291).
The formula I compounds can be prepared using the reactions described in methods A-E.
Possible COX hydrolysis reactions, where X coincides with what is specified in the description of the general formula, for preparing the corresponding free acids, can be conducted according to the common laboratory procedures for this type of reaction.
The meaning of the various symbols, unless otherwise specified, is intended to coincide with the indications provided in the general formula.
METHOD A
Step 2 Mitsunobu ■"-θ-' R1 R2 l, 1
Step 3
Catalytic hydrogenation
I (Y = 0, -CH2 )
The preparation of general formula I compounds was done by reacting a general formula II compound with a general formula III compound with a base, preferably inorganic, and preferably sodium hydride, to form the corresponding intermediate product IV, which was then reacted with a formula II compound in the classic Mitsunobu reaction conditions, as described in Synthesis 1981, 1-28, using anhydrous and aprotic solvents such as benzene, toluene, ether or preferably tetrahydrofuran, for time periods ranging from 30 minutes to 72
hours, preferably 18 hours, at temperatures ranging from 10 to 40°C, preferably 25°C.
Product V obtained could then be subjected to catalytic hydrogenation in the presence of H2, at a pressure ranging from atmospheric pressure to 60 psi, preferably 50 psi, and with catalysts such as metals supported on C, such as Pd/C, in percentages ranging from 1 to 20%, preferably 10%. The amount of catalyst used was within the 1- 100% p/p range, usually 10% p/p, in protic or non-protic solvents, such as MeOH, dioxane and THF, preferably MeOH, for reaction times ranging from 18 hours to 3 days, preferably 24 hours.
METHOD B
Step l Y = -CH=
VI VIII
Step 2
Catalytic hydrogenation
R1 = H (see method A
R2 = COX step 3
Z, = Z2 = 1
X other than OH
Y = -GH2
I
The general formula I compounds were synthesised starting from general formula VI compounds dissolved in aprotic solvents
such as toluene, and reacted with a general formula VII compound, at reflux temperature with Dean-Stark, for time periods ranging from 5 to 24 hours, preferably 7 hours, in the presence of a catalysts such as a salt of an organic base with an organic acid, such as piperidine acetate, normally used in Knoevenagel reactions. Alternatively, the reaction was conducted in dipolar aprotic solvents such as DMF (Synthetic Communication, 2000, 30 (4), 713-726) possibly in the presence of an organic base such as piperidine, at temperatures ranging from 20 to 100°C, preferably 80°C for time periods ranging from 1 hour to 3 days, preferably 2 days. Compound VIII was then subjected to catalytic hydrogenation in the conditions and with the times described in method A (step 3) to yield the general formula I product. METHOD C
Ste l Base "
IX
XI (Y = -CH=)
VII
I A (Y = -CH2) I B ( Y = -CH2 )
( n = 2 - 4 unsaturated) ( n = 1 - 4 saturated )
X other than OH n=1-4
L = leaving group, preferably halogen, preferably Br
R, = H
Z, = Z2=1
The preparation of general formula I A and I B compounds was done by reacting two equivalents of a general formula II compound with one equivalent of a formula IX compound with a base, preferably inorganic, and preferably sodium hydride, to form the corresponding intermediate product X.
Intermediate product X was then reacted with a general formula VII compound, in the conditions described in general method B (step 1), to form the intermediate general formula XI product, which in turn was reacted with a general formula VII compound, in the conditions described in method B to form the intermediate product XII. Intermediate product XII was then subjected to reduction with metal hydrides, for example, sodium borohydride in a polar protic solvent, preferably MeOH, for a time period ranging from 2 to 24 hours, preferably 18 hours, to give compound I A. Product I B, on the other hand, was obtained by reduction with metal supported in a hydrogen atmosphere according to the specifications of general method A (step 3) .
If the [CR3R4]n group does not present unsaturations, IA and IB are the same; if the [CRsR4]n group presents unsaturations IA and IB are not the same.
METHOD D
XII
XIV
X other than OH γ = o
L = leaving group Z, = Z2=1
The preparation of general formula I (and general formula XIV) compounds was done, for example, according to the description in Tetrahedron, 1990, 46 (3), 967-978 starting from product XII which was reacted with a general formula XIII compound containing a leaving group (for example, chlorine, bromine, iodine, mesyl, tosyl), for example, methyl-2-bromoisobutyrate in the presence, alternatively, of base alone, preferably sodium hydride, in polar aprotic solvents, preferably DMF at temperatures ranging from 25°C to the reflux temperature of the solvent selected or of a base, such as potassium carbonate, and a catalyst by phase transfer, such as, for example, tetrabutylammonium bromide (TBAB) in aprotic solvents such as toluene, at temperatures ranging from 25°C to the reflux temperature of the solvent selected, preferably the reflux temperature, for time periods ranging from 1 to 5 days, preferably 3 days. The two compounds obtained were separated using a chromatographic method, preferably chromatography on a silica gel column, using eluotropic mixtures of varying polarity from pure hexane to pure ethyl acetate, preferably mixtures of hexane/ ethyl acetate, in varying ratios to one another, preferably 8/2.
METHOD E
XVII
XV step 1
X other than OH
The preparation of the general formula I compound was done by reacting a general formula XV compound with a general formula XVI compound in the presence of dimeric rhodium (II) acetate as the catalyst to form general formula compound XVII. Compound XVII was then reacted again (with a general formula XVI compound, in the same conditions as described above to yield general formula compound I, in a polar solvent such as acetonitrile, or preferably toluene, for a time period ranging from 18 to 48 hours, preferably 24 hours, at a temperature ranging from 10 to 130°C, preferably reflux temperature.
EXAMPLE 1
Preparation of dimethyl 2-[4-(2-{4-[3-methoxy-2-
(methoxycarbonyl)-3-oxopropyl]fenoxy}ethoxy)benzyl1malonate (ST1720)
Preparation of the intermediate product dimethyl 4-|~(2- hydroxyethyD-oxylbenzylidenemalonate
Sodium hydride (244 mg, 10.16 mmol) was added to dimethyl 4-hydroxybenzylidenemalonate (2 g, 8.47 mmol) in 40 mL of anhydrous DMF. The suspension was left under magnetic stirring for 30 minutes until solution was complete, and then 2- bromoethanol (0.780 mL, 11.01 mmol) was added. The solution was left overnight under magnetic stirring at room temperature. The
reaction was processed, diluting with AcOEt and washing with water. The organic phase was dried on anhydrous Na2S0 , and, after filtration, the solvent was evaporated in vacuo. The residue was purified by chromatography on silica gel using a hexane/AcOEt gradient in a ratio of 8:2 to 6:4 as the eluent. 1.760 g of product were obtained (yield: 74%); iHNMR (CDCI3, 300 MHz) δ 7.80 (s, 1H), 7.40 (d, 2H), 6.90 (d, 2H), 4.20 (brd, 2H), 4.00 (brd, 2H), 3.80 (d, 6H).
Preparation of the intermediate product l ,2-bis[4-(methylene- dicarboxylate dimethyl) phenoxyl ethane
Method A (step2)
To dimethyl 4-[(2-hydroxyethyl)-oxy]benzylidenemalonate (1.760 g, 6.28 mmol) (prepared as described above) dissolved in 50 mL of THF were added 4-hydroxybenzylidenedimethylmalonate (1.482 g, 6.28 mmol), PPh3 (2.138 g, 8.16 mmol), and DEAD (1.28 mL, 8.16 mmol dissolved in 10 mL of THF). The solution thus obtained was left
under magnetic stirring for three days at room temperature. After this time, the formation of an insoluble product was observed, which by filtration yielded 1.3 g of product which was used as is for the next reaction (yield: 41.5%); *HNMR (CDCI3, 300 MHz) δ 7.80 (s, 2H), 7.40 (d, 4H), 6.90 (d, 4H), 4.40 (s, 4H), 3.80 (d, 12H).
Preparation of dimethyl 2- -(2-{4-[3-methoxy-2-
(methoxycarbonyl)-3-oxopropynphenoxy}ethoxy)benzyllmalonate (ST 1720)
1.3 g of l,2-bis[4-(methylenedicarboxylate dim ethyl) phenoxy]- ethane were solubilised in 70 mL of MeOH and 240 mg of 10% Pd/C were added; the suspension was subjected to catalytic hydrogenation at room temperature and at 60 psi hydrogen pressure. The mixture was filtered in celite and the filtrate was washed thoroughly with CHCI3. 1,280 g of product were obtained and purified by chromatography on silica gel using a hexane/AcOEt gradient from 80:20 to 65:35 as the eluent. 460 mg of product were obtained (yield: 35%); Mp (melting point): 90°C; TLC: silica gel; eluent; hexane/AcOEt 1/ 1 Fr (frontal ratio): 0.55; *HNMR (CDCI3,
300 MHz) δ 7.10 (d, 4H), 6.80 (d, 4H), 4.30 (s, 4H), 3.70 (s, 12H), 3.60 (t, 2H), 3.25 (d, 4H); HPLC: Column: Inertisil ODS-3 (5μm), 4.6 x 250 mm, T: 30°C: mobile phase: CH3OH/KH2PO4 50 mM (65/35 v/v); flow rate: 1 mL/min; UV 205 nm detector, retention time: 22.69 min; E.A.: conforms for C26H30O10. EXAMPLE 2
Preparation of dimethyl 2-{4'-r3-methoxy-2-
(methoxycarbonyl)-3-oxo- 1 -propenyl] [ 1 , 1 '-biphenyl -4-yl|- 1, 1- ethylenedicarboxylate (ST2013)
Method B (stepl)
To a solution of 4,4'-bis benzaldehyde (2 g, 9.51 mmol) in 30 mL of anhydrous DMF were added dimethyl malonate (3.76 g, 28.53 mmol) and piperidine (0.121 g, 1.425 mmol). The solution was heated to 80°C and left under magnetic stirring for 48 hours, and then poured into water and extracted three times with CHCI3. The pooled organic phases were dried on anhydrous sodium sulphate
and the solvent evaporated in vacuo. The residue obtained was purified by chromatography on silica gel, eluting with hexane/AcOEt 8/2. 550 mg of product were obtained (yield: 13%); Mp: 163°C; TLC: silica gel; eluent: hexane/AcOEt 6/4, Fr: 0.43; ΗNMR (CDCI3, 300 MHz) δ 7.80 (s, 2H), 7.60 (d, 4H), 7.50 (d, 4H), 3.90 (d, 12H); HPLC: Column: Symmetry-Cis (3.5 μm); mobile phase: CH3CN/H2O 50/50 v/v, pH: as is; flow rate: 0.9 mL/min; temperature: RT; UV 205 nm detectors, retention time: 10.7 min; E.A.: conforms for C24H22O8.
EXAMPLE 3
Preparation of dimethyl 2-({4'-f3-methoxy-2-(methoxy- carbonyl)-3-oxopropyT| [1 , 1 '-biphenvH-4-yl)methyl)nιalonate (ST 2032)
Method B (step 2)
To a solution of ST 2013 (550 mg, 1.24 mmol) (prepared as described in example 2) in 50 mL of THF were added 55 mg of 10% Pd/C. The suspension was subjected overnight to catalytic hydrogenation at room temperature and 60 psi hydrogen pressure.
After filtration of the mixture on celite, the solvent was evaporated in vacuo and the residue purified by chromatography on silica gel, eluting with hexane/ AcOEt 8:2. 160 mg of product were obtained (yield: 30%); Mp: 128.5°C; TLC: silica gel; eluent: hexane/AcOEt 6:4, Fr: 0.58; iHNMR (CDCla, 300 MHz) δ 7.50 (d, 4H), 7.25 (d, 4H), 3.75 (s, 12H), 3.25 (d, 4H); HPLC Column: Inertisil - ODS 3 (5 μM); mobile phase: CH3CN/H2O 70/30 v/v; temperature: RT; pH: as is; flow rate: 0.75 mL/min; UV 205 nm detectors, retention time 10.82 min; E.A. : conforms for C24H26O8.
EXAMPLE 4
Preparation of dimethyl 2-{4-[((2Z)-4-{4-[3-methoxy-2- (methoxy-carbonyl)-3-oxo- l-propenyl|phenoxyl-2-butenyl)oxy1- phenyl}- 1 , 1 -ethylenedicarboxylate(ST2012)
Preparation of intermediate product 1 ,4 dibromo-2-cis-butene
The preparation was done according to the procedure described in Synth. Commun. 1991, 721-726.
Preparation of the intermediate product 1 ,4-bis (4- formylphenoxy) -2-cis-butene
To a solution of 4-hydroxybenzaldehyde (1 g, 8.18 mmol) in 15 mL of anhydrous DMF was added NaH (258 mg, 10.87 mmol). The suspension was left under magnetic stirring for 10 minutes at room temperature. After solubilisation, l ,4-dibromo-2-cis-butene (874 mg, 4.09 mmol) was added. The reaction was left overnight under magnetic stirring at room temperature, the solvent was then evaporated in vacuo and the semisolid obtained was treated several times with hexane until a dark solid was obtained. The residue was purified by chromatography on silica gel, eluting with hexane/AcOEt 8:2. 775 mg of product were obtained (yield: 32%); iHNMR (CDCI3, 300 MHz) δ 9.90 (s, 2H), 7.90 (d, 4H), 7.00 (d, 4H), 6.00 (m, 2H), 4.80 (s, 4H) .
Preparation of the intermediate product dimethyl 2-(4-{|
~(2Z)- 4-(4-formylphenoxy)-2-butenyl]oxy}benzylidene)malonate
The title product was prepared according to the procedure described in Method B step 1, adding dimethylmalonate (0.338 g, 2.56 mmol), piperidine (0.032 g, 0.38 mmol) and acetic acid (0.023 g, 0.38 mmol) to a solution of 1,4-bis (4-formylphenoxy6)-2-cis-butene (380 mg, 1.28 mmol) in 30 mL of toluene. The solution was refluxed for 5 hours at 140°C, and then the solvent was evaporated in vacuo and the crude product obtained was purified by chromatography on silica gel, eluting with hexane/ethyl acetate 8:2. 320 mg of product were obtained (yield: 60 %); ΗNMR (CDCI3, 300 MHz) δ 9.90 (s, IH), 7.90 (d, 2H), 7.70 (s, IH), 7.40 (d, 2H), 7.10 (d, 2H), 6.90 (d, 2H),
6.00 (m, 2H), 4.70 (m, 4H), 3.90 (d, 6H).
Preparation of dimethyl 2-(4-[((2Z)-4-{4-f3-methoxy-2- (methoxy- carbonyl) - 3 - oxo- 1 - propenyll phenoxyj- 2 - butenyl) oxy) f enylj-
1.1 -ethylenedicarboxylate (ST2012)
The title product was prepared according to the procedure described in Method B step 1, adding dimethylmalonate (0.080 g, 0.610 mmol), piperidine (0.013 g, 0.152 mmol) and acetic acid
(0.009 g, 0.152 mmol) to a solution of dimethyl 2-(4-{[(2Z)-4-(4- formylphenoxy)-2-butenyl]oxy}benzylidene) malonate (320 mg, 0.610 mmol) in 35 mL of toluene. The mixture thus obtained was refluxed for 5 hours at 140°C, and then the solvent was evaporated in vacuo and the crude product obtained was purified by chromatography on silica gel, eluting with CHCI3. 250 mg of product were obtained (yield: 78%); Mp: 80-82°C; TLC: silica gel; eluent: CHCI3, Fr: 0,41; iHNMR (CDCI3, 300 MHz) δ 7.70 (s, 2H), 7.40 (d, 4H), 6.90 (d, 4H), 5.95 (t, 2H), 4.70 (d, 4H), 3.85 (d, 12H); HPLC Column: Symmetry - Ci8 (3.5 μm), mobile phase: CH3CN/H2O 50/50 v/v; pH: as is; temperature: RT, flow rate: 0.9 mL/min, retention time: 18.51 min; E.A. : conforms for C28H28O10.
EXAMPLE 5
Preparation of dimethyl 2-{4-[((2Z)-4-{4-[3-methoxy-2- (m.ethoxycarbonyl)-3-oxopropyl1phenoxyl-2-butenyl)oxy|benzyl|- malonate (ST2145)
Method C (step 2)
To a solution of ST 2012 (2.4 g, 4.57 mmol) (prepared as described in example 4) in 50 mL of anhydrous methanol was added
NaBH
4 (0.451 g, 1 1.92 mmol). The suspension was left under magnetic stirring for 40 minutes at room temperature. The mixture was then poured into ethyl acetate and the organic phase washed with water and then dried on anhydrous sodium sulphate. The solvent was evaporated in vacuo and the residue purified by chromatography on silica gel (previously treated with triethylamine) , eluting with hexane/ AcOEt 8:2. 0.460 g of product were obtained (yield: 19%); TLC: silica gel; eluent: hexane/ AcOEt 6:4, Fr: 0.49;
iHNMR (CDCI3 300 MHz) δ 7.01 (d, 4H), 6.80 (d, 4H), 5.95 (t, 2H), 4.64 (d, 4H), 3.70 (s, 12H), 3.62 (t, 2H), 3.15 (d, 4H); HPLC: Column: Inertisil - ODS 3 (5 μM), 4.6 x 250 mm; mobile phase: CH3CN/H2O 70/30 v/v, temperature: RT; pH: as is; flow rate: 0.9 mL/min; UV 205 nm detectors, retention time 11.61 min; E.A. : conforms for
EXAMPLE 6
Preparation of dimethyl 2-[4-(4-{4-[3-methoxy-2-(methoxy- carbonyl)-3-oxopropyHphenoxy}butoxy)benzvHmalonate (ST2144)
Method C (step 2')
To a solution of ST2012 (1.85 g, 3.52 mmol) (prepared as described in example 4) in 150 mL of THF were added 230 mg of 10% Pd/C. The suspension was subjected overnight to catalytic hydrogenation at room temperature and at 60 psi hydrogen pressure. After filtering the mixture on celite, the solvent was evaporated in vacuo and the residue purified by chromatography on silica gel, eluting with hexane/ AcOEt 8:2. 940 mg of product were obtained (yield: 49); Mp: 83°C; TLC: silica gel; eluent: hexane/AcOEt 6:4, Fr: 0.53;
iHNMR (CDC1
3? 300 MHz) δ 7.05 (d, 4H), 6.80 (d, 4H), 4.00 (t, 4H), 3.70 (s, 12H), 3.60 (t, 2H), 3.15 (d, 4H); HPLC Column: Inertisil - ODS 3 (5 μM), 4.6 x 250 mm; mobile phase: CH3CN/H2O 70/30 v/v, temperature: RT; pH: as is; flow rate: 0.75 mL/min, UV 205 nm detectors, retention time 13.30 min; E.A. conforms for
EXAMPLE 7 AND EXAMPLE 8
Preparation of methyl 2-{3-[2-(3-hvdroxyphenoxy)ethoxy1- ρhenoxyl-2-methylproρanoate (ST1877) and methyl 2-(3-{2-[3-(2-
methoxy- 1 , l-dimethyl-2-oxoethoxy)phenoxy]ethoxy}phenoxy)-2- methylpropanoate (ST 1878)
ST 1877
ST 1878
Method D
A mixture of 3,3'-ethylenedioxidiphenol (2.000 g, 8.1 mmol), K2CO3 (4.500 g, 32.4 mmol), TBAB (0.131 g, 0.4 mmol) and methyl- 2-bromoisobutyrate (11.611 g, 64 mmol) in 100 mL of toluene was heated at 130°C for three days, and then cooled and filtered. The solid obtained was washed with toluene, the pooled organic phases were evaporated in vacuo and the oily residue was purified by chromatography on silica gel using hexane/AcOEt 8:2 as the eluent. Two products were obtained: the monoderivative ST1877 (0.700 g; yield: 25%) and the biderivative ST1878 (1.100 g; yield: 30.4 %).
Analytical data for ST 1877 (monoderivative, Example 7)
Mp: 77-79°C; *HNMR (CDCI3, 300 MHz) δ 7.13 (t, 2H), 6.62 - 6.40 (m, 6H), 4.25 (s, 4H), 3.78 (s, 3H), 1.60 (s, 7H); HPLC: Column:
Inertisil ODS - 3 (5 μm), 4.6 x 250 mm, T: 30°C, mobile phase:
CH3CN/H2O (60/40 v/v), pH: 3.2, flow rate: 1.0 mL/min, UV 205 nm detector, retention time: 8.76 min; E.A. conforms for C19H22O6.
Analytical data for ST 1878 (biderivative, Example 8)
Mp: 60-62°C; ΗNMR (CDC13, 300 MHz) δ 7.13 (t, 2H), 6.60 (d, 2H), 6.41 (m, 4H), 4.26 (s, 4H), 3.78 (s, 6H), 1.60 (s, 12H); HPLC: Column: Inertisil ODS - 3 (5 μm), 4.6 x 250 mm, T: 30°C; mobile phase: CH3CN/H2O (60/40 v/v), pH: 3.2, flow rate: 1.0 mL/min; UV 205 nm detector, retention time: 23.92 min; E.A.: conforms for
EXAMPLE 9
Preparation of dimethyl 2-{4-[l-(4-hydroxyphenyl)- l- methylethyll fenoxyjmalonate (ST2020)
To a suspension of rhodium diacetate (0.220 g, 0.5 mmol) and bisphenol A (3.400 g, 15 mmol) in 100 mL of anhydrous toluene was added dropwise, under nitrogen flow, a solution of diazomalonate (3.000 g, 18 mmol, prepared as described in Org. Synth: 1973, V, 179) in 100 mL of anhydrous toluene, taking care to keep the temperature during the addition between 15 and 20°C. The reaction mixture was then refluxed at 120- 130°C for 24 hours under nitrogen. After this time period the mixture was filtered and the toluene evaporated in vacuo. The residue obtained was purified by means of chromatography on silica gel using hexane/AcOEt 8:2 as
the eluent. 1.700 g of oil product were obtained (yield: 32%); TLC: silica gel; eluent hexane/AcOEt 7:3, Fr: 0.23; *HNMR (CDCI3, 300 MHz) δ 7.16 (m, 4H), 6.90 (d, 2H), 6.87 (d, 2H), 5.12 (s, IH), 3.90 (s, 6H), 1.62 (s, 6H); HPLC: Column: Inertisil ODS - 3 (5 μm) 4.6 x 250 mm T: 30°C; mobile phase: CH3CN/H2O 70/30 (v/v), pH: as is, flow rate: 0.75 mL/min; UV 205 nm detector, retention time 7.0 min; E.A. conforms for C20H22O6.
EXAMPLE 10
Preparation of dimethyl 2-[4-(l-{4-[2-methoxy- l- (methoxycarbonyl)-2-oxoethoxy]phenyl|- 1 -methylethyl)- phenoxylmalonate (ST2048)
Method E (step 2)
The product was prepared starting from rhodium diacetate (0.885 g, 0.2 mmol) and ST 2020 (1.230 g, 3.4 mmol) (prepared as described in example 9) in 36 mL of anhydrous toluene, adding diazomalonate (1.882 g 11.9 mmol of preparation as described in Org. Synth: 1973, V, 179) dropwise in 18 mL of anhydrous toluene, taking care to keep the temperature between 15 and 20°C. The reaction mixture was refluxed at 120- 130°C for 24 hours under nitrogen. The mixture was filtered and the toluene evaporated in
vacuo. The residue obtained was purified by chromatography on a silica gel column using hexane/ AcOEt 8:2 as the eluent. 0.430 g of oily product were obtained (yield: 26%); TLC: silica gel; eluent: hexane/AcOEt 6:4, Fr: 0.46; *HNMR (CDCI3, 300 MHz) δ 7.20 (d, 4H), 6.90 (d, 4H), 5.22 (s, 2H), 3.90 (s, 12H), 1.61 (s, 6H); HPLC: Column: Inertisil ODS - 3 (5 μm) 4.6 x 250 mm, T: 30°C; mobile phase: CH3CN/H2O 70/30 (v/v), pH: as is, flow rate: 0.75 mL/min, UV 205 nm detector, retention time 9.68 min; KF: 0.7 % H20; E.A. conforms for C25H28O10.
EXAMPLE 11
Preparation of methyl 2-{[4'-(2-methoxy- l , l-dimethyl-2- oxoethoxy) [ 1 , 1 '-biphenyl|-4-ylloxy|-2-methylpropanoate (ST2291 )
Method D
The title product was prepared starting from a solution of 4,4' bisphenol (0.5 g, 2.68 mmol), in 25 mL of anhydrous DMF, to which were added sodium hydride (0.321 g, 10.72 mmol), and after 10 minutes at room temperature methyl-2-bromoisobutyrate (1.06 g, 5.89 mmol). The mixture was left under magnetic stirring for 3 days at room temperature, and then poured into ethyl acetate. The
organic phase was washed with a 5% NaOH solution and then with water, and then dried on anhydrous sodium sulphate, filtered and evaporated in vacuo. The residue was purified by chromatography on a silica gel column, eluting with hexane/AcOEt 8/2. 0.175 of product were obtained (yield: 18%) Mp: 58-59°C; TLC: silica gel; eluent: hexane/AcOEt 7:3, Fr: 0.6; *HNMR (CDCls, 300 MHz) δ 7.40 (d, 4H), 6.85 (d, 4H), 3.80 (s, 6H), 1.65 (s, 12H); HPLC: Column: Symmetry Ciβ 4.6 x 150 mm (5 μm), T: RT; mobile phase: CH3CN/H2O 60/40 (v/v), pH: as is; flow rate: 0.8 mL/min; UV 205 nm detector, retention time 16.48 min; KF: 0.75% H2O; E.A. conforms for C22H26O6.
The compounds according to the present invention are useful as medicines, and particularly for the preparation of medicines with serum glucose and serum lipid lowering activity. Preferred applications are the prophylaxis and treatment of diabetes, particularly type 2 diabetes and its complications, X syndrome, the various forms of insulin resistance and hyperlipidaemias.
In a thoroughly advantageous manner, the compounds according to the present invention are endowed with good pharmacological activity and present reduced liver toxicity. EXAMPLE 12 Antidiabetic and serum lipid lowering activity in db/db mice
Mutations in laboratory animals have made it possible to develop models that present non-insulin-dependent diabetes
associated with obesity, hyperlipidaemia and insulin resistance and that make it possible to test the efficacy of new antidiabetic compounds (Reed and Scribner, Diabetes, obesity and metabolism 1: 75 - 86, 1999). A genetically diabetic mouse model much used in the laboratory is the C57BL/KsJ db/db mouse.
The genetic basis of this model is a defect in the leptin receptor gene that determines leptin resistance and leads to hyperphagia, obesity, hyperinsulinaemia and insulin resistance, with subsequent symptoms of insufficient insular secretion and hyperglycaemia (Kodama et al, Diabetologia 37: 739 - 744, 1994; Chen et al, Cell 84: 491 - 495, 1996).
Since hyperglycaemia is accompanied by obesity and insulin resistance, the db/db mouse model presents characteristics that cause it to resemble type 2 diabetes in man and is useful for assaying insulin-sensitising compounds.
One class of these compounds consists in the thiazolidinediones (Day, Diabet. Med. 16: 179-192, 1999; Mudaliar and Herry, Annu. Rev. Mred. 52: 239 - 257, 2001, Drexler et al, Geήatrix 56: 20 - 33, 2001).
Of the three thiazolidinediones launched on the market, troglitazone was withdrawn owing to serious liver toxicity, while the other two compounds, rosiglitazone and pioglitazone, are effective in reducing diabetic hyperglycaemia despite presenting unwanted side
effects (Schoonjans and Auwerx, The Lancet 355: 1008 - 1010, 2000; Peters, Am. J. Manag. Care 7: 587-595, 2001; Gale, The Lancet 357: 1870 - 1875, 2001).
The C57BL/KsJ db/db mice used in the experiments were supplied by Jackson Lab (via Ch. River) . After 10 days' acclimatisation in standard conditions (22 ± 2°C; 55 ± 15% humidity; 15 - 20 air changes/hour; 12 hour light-dark cycle with light from 7.00 a.m. to 7 p.m.) on a standard 4 RF21 diet (Mucedola), blood samples were taken in post- absorption conditions (fasting from 8.30 a.m. to 4.30 p.m.) from the caudal vein with the aid of a Jelco 22G catheter (Johnson and Johnson). Levels of glucose, triglycerides and cholesterol were measured in plasma for a well matched distribution of the mice in the treatment groups.
The body weight of the animals was checked at the start of treatment and the monitoring of water and animal feed consumption was arranged.
The mice were treated twice daily (8.30 a.m. and 6.30 p.m.), orally, for 11 days.
The compounds were administered at the dose of 35 mg/kg [ST2145 (example 5)], in 10 ml/kg of vehicle (1% CMC containing
Tween 80 0.5% in deionised H20). The reference compound, rosiglitazone, was administered at the dose of 5 mg/kg (Lohray et al.
J. Med Chem 41, 1619 - 1630, 1998).
The animals were sacrificed (by decapitation) in post- absorption conditions (fasting from 9.30 a.m. to 4.30 p.m.), 7 hours after the last treatment. Serum levels of a number of important lipid and carbohydrate metabolism parameters were determined. 5 The compounds according to the invention are capable of lowering serum glucose levels, of reducing weight gain and of reducing the production of transaminase (GPT), which is an indicator of less liver damage. By way of an example, we give the serum glucose lowering activity and the changes in weight and lo transaminase levels for ST2145 compared to rosiglitazone.
The results obtained are presented in Table 1. TABLE 1
Serum glucose and serum lipid lowering activity, and variations in serum GPT levels in db/db mice, and weight gain, after is 11 da s' treatment
Student's 't'-test: A indicates P < 0.001; Δ indicates P < 0.01; ■ indicates P < 0.02 vs. Controls.
Also objects of the present invention are pharmaceutical compositions containing as their active ingredient at least one
formula (I) compound, alone or in combination with one or more formula (I) compounds, or, said formula (I) compound or compounds in combination with active ingredients useful for the treatment of the diseases indicated in the present invention, for example, other products with serum glucose and serum lipid lowering activity; also in separate dosage forms or in forms suitable for combined therapies. The active ingredient according to the invention will be in a mixture with suitable vehicles and/or excipients commonly used in pharmacy, such as, for example, those described in "Remington's Pharmaceutical Sciences Handbook", latest edition. The compositions according to the present invention will contain a therapeutically active amount of the active ingredient. The doses will be determined by the expert in the sector, e.g. the clinician or primary care physician, according to the type of disease to be treated and the patient's condition, or concomitantly with the administration of other active ingredients. By way of an example, doses ranging from 0.1 to 200 mg/day may be indicated.
Examples of pharmaceutical compositions are those that permit oral or intravenous, intramuscular, subcutaneous, or transdermal parenteral administration. Pharmaceutical compositions suitable for the purpose are tablets, rigid or soft capsules, powders, solutions, suspensions, syrups, and solid forms for extempore liquid preparations. Compositions for parenteral administration are, for example, all the intramuscular, intravenous,
subcutaneous injectable forms, in the form of solutions, suspensions, or emulsions. We should also mention the liposomal formulations. Also included are the forms with controlled release of the active ingredient, whether as oral administration forms, tablets coated with appropriate layers, microencapsulated powders, complexes with cyclodextrins, depot forms, for example, subcutaneous ones, such as depot injections or implants.