MXPA00010998A - Novel fatty analogues for the treatment of diabetes - Google Patents

Abstract

The present invention relates to novel fatty acid analogues of the general formula (I):CH3-[CH2]m-[xi-CH2]n-COOR, as defined in the specification, which can be used for the treatment and/or prevention of diabetes. Further, the invention relatesto a nutritional composition comprising such fatty acid analogues.

Description

ANALOGUES OF FATS FOR THE TREATMENT OF DIABETES FIELD OF THE INVENTION The present invention relates to novel fatty acid analogs that can be used for the treatment and / or prevention of diabetes. In addition, the invention relates to a nutritional composition comprising such fatty acid analogues.

BACKGROUND OF THE INVENTION Diabetes mellitus and its complications are now considered the third leading cause of death in Canada and the United States, behind only cancer and cardiovascular disease. The treatment with modified fatty acids represents a new way to treat these diseases. EP 345,038 and PCT / N095 / 00195 disclose the use of non-β-oxidizable fatty acid analogues.

It has now been found that they have a wider area of applications. In addition, the new fatty acid analogues have been synthesized and characterized now, which impose an effect on diabetes. In feeding experiments with fatty acid, the results show that these compounds decrease adipose tissue mass and body weight and are thus potent drugs for the treatment of obesity and overweight. In addition, it has now been shown that fatty acid analogues are potent antidiabetic compounds, with a profound effect on glucose and insulin levels. In addition, it has been proven that the compounds have a favorable effect on restenosis, and show good antioxidant properties.

DIABETES Diabetes mellitus and its complications are now considered the third leading cause of death in Canada and the United States, behind only cancer and cardiovascular disease. Although the acute and often lethal symptoms of diabetes can be controlled by insulin therapy, long-term complications reduce life expectancy by as much as a third. Compared with incidence rates in normal non-diabetic people, diabetic patients show rates that are increased 25 times for blindness, 17 times for kidney disease, 5 times for gangrene, and 2 times for heart disease. There are two major forms of diabetes mellitus. One is type I diabetes, which is also known as insulin-dependent diabetes mellitus (IDDM), and the other is type II diabetes, which is also known as diabetes mellitus II, not insulin-dependent (NIDDM). Most patients with IDDM have a common pathological picture: the almost complete disappearance of pancreatic beta-producing insulin cells which results in hyperglycemia. Considerable evidence has accumulated that shows that most IDDM is the consequence of the progressive destruction of beta cells during an asymptomatic period that frequently extends: in many years. The pre-diabetic period can be recognized by the detection of autoantibodies against islet cells, circulating cells, and autoantibodies against insulin. There is a need for a compound that could be non-toxic and have no side effects, but could prevent IDDM and NIDDM clinics. Type I diabetes: severe diabetes mellitus, usually of abrupt onset before maturity, characterized by low levels of plasma insulin, polydipsia, polyuria, increased appetite, weight loss and ketoacidosis in episodes; also called IDDM. Type II diabetes: a frequently mild form of diabetes mellitus, often of gradual onset, usually in adults, characterized by normal to high plasma insulin levels that are relatively low in relation to plasma glucose levels; also called NIDDM. Type I and II diabetes are in accordance with an etiological classification considered as "primary" diabetes respectively.

Secondary diabetes comprises pancreatic diabetes, ext. pancreatic / endocrine or drug induced. In addition, some types of diabetes are classified as exceptional forms. These include lipoatrophic, miatonic diabetes and a type of diabetes caused by disturbance of insulin receptors. Considering the high occurrence of diabetes in our society and the serious consequences associated with it as discussed above, any therapeutic drug potentially useful for the treatment and prevention of this disease could have a profound beneficial effect on your health. There is a need in the art for a drug that reduces the concentration of glucose in the blood of diabetic subjects without significant adverse side effects. It is therefore an object of the present invention to provide a treatment regimen that is useful for lowering blood glucose and for treating a diabetic condition. Yet another objective of the present invention is to provide a treatment regimen that is useful for decreasing the concentration of insulin in the blood, and for increasing the effect of the remaining insulin.

ACTION MECHANISMS Minor modifications of natural fatty acids, sulfur, selenium or oxygen, replace one or more of the carbon atoms in the main chain of the fatty acid. The compounds defined by formula I have properties that give them a unique combination of biological effects. Tetradecylthioacetic acid (TTA) is more fully studied and has been shown to have several beneficial effects in various animal tests. Studies have shown that TTA has properties very similar to natural fatty acids, the main difference being that TTA is not oxidized by the mitochondrial ß-oxidation system. However, the presence of the compounds of the present invention has not been shown to increase the β-oxidation of the others (unsubstituted fatty acids). The administration of TTA to rats for 12 weeks almost doubled the hepatic and plasma content of the monounsaturated fatty acids (mainly oleic acid), whereas the polyunsaturated fatty acids (mainly linoleic acid and DHA) decreased. In this way the compound of the present invention modifies the composition of the lipids in various tissues. It was also shown that the present compounds modify the fatty acid content, and it was anticipated that the present compounds also modify the distribution of fats. Moderate doses of TTA feed to animals such as rats, mice, rabbits and dogs decreased plasma cholesterol and triacylglycerol levels within a few days of treatment. The same effect has also been shown for TSA, and compounds of the present invention with sulfur substituted at positions 5 or 7, which have been shown to increase β-oxidation and thus it is anticipated that also these fatty acid analogues will decrease the plasma levels of triglycerides and cholesterol. TTA and TSA are much more potent in this respect than polyunsaturated fatty acids such as EPA.

As mentioned above, an important mechanism of action of type 3-thia fatty acids is a significantly increased oxidation of mitochondrial fatty acid, which reduces the availability of fatty acids for esterification. The synthesis of cholesterol and cholesterol is reduced and the VLDL secretion of the liver is reduced (10). This has the effect of reducing the production of LDL. All these effects seem to be at least partially mediated by the receptors]: it is activated by the peroxisome proliferator (PPAP), ubiquitous transcription factors involved in the regulation of lipid metabolism. We have shown that TTA is a potent PPARα ligand, a transcription factor that regulates the catabolism of fatty acids and the cosanoids, and a less potent PPARα ligand, which is involved in the regulation of the differentiation of adipocytes. Obesity is a common feature of non-insulin-dependent diabetes mellitus (NIDDM) and a risk factor for its development. NIDDM is frequently linked to hypertension, dyslipidemia, high levels of plasma free fatty acids and an increased risk of cardiovascular disease. Patients with NIDDM are characterized by resistance to the action of insulin on glucose uptake in peripheral tissues and poorly regulated insulin secretion. We have shown that TTA decreases hyperinsulinemia and markedly improved the action of insulin on the utilization of glucose. TTA also prevented insulin resistance induced by diet. In contrast to the previously known antidiabetic glitazones, TTA did not increase the increase in body weight. These effects can be at least partially explained by the increased influx of fatty acids and the increased oxidation of the fatty acid in the liver. The data thus suggest a role for TTA in the homeostasis of lipids and glucose in vivo. As clearly shown in the experimental section, the compounds of the present invention inhibit an increase in body weight and adipose tissue mass of animals given either a high fat or high sucrose content diet. This makes the compounds of the present invention very suitable as pharmaceutical and / or nutritional agents for the treatment of obesity, for example compounds that can be used as a slimming agent to provide a reduction in body weight or weight of adipose tissue In addition, the compounds of the present invention can be used as an antidiabetic drug by reducing the concentration of glucose in the blood. It has also been shown that the compounds of the present invention reduce the plasma concentration of insulin in hyperinsulinemic animals. For animals who have reduced sensitivity to insulin, the compounds of the present invention have been shown to reinforce the effect of endogenous insulin. The term "metabolic syndrome" is used to describe a multimetabolic syndrome which is inter alia characterized by hyperinsulinemia, insulin resistance, obesity, glucose intolerance, type 2 diabetes mellitus, dyslipidemia or hypertension. As indicated above, the compounds of the present invention have been shown to provide a positive effect on all the aforementioned conditions, for example in regulating the homeostasis of glucose and lipids, and thus it is anticipated that the compounds of the present invention will be suitable agents for the regulation of the metabolic disease defined above (sometimes called syndrome X).

DETAILED DESCRIPTION OF THE INVENTION The present invention discloses that modified analogs of fatty acid at non-cytotoxic concentrations can be used for the treatment and / or prevention of obesity, hypertension and fatty liver. The present invention relates to the use of fatty acid analogues of the general formula (I): CH3- [CH2] 3, - [X? -CH2) n-COOR where n is an integer from 1 to 12; and where m is an integer from 0 to 23, and where i is an odd number indicating the position relative to COOR, and where X, independently of one another, is selected from the group comprising 0, S, SO, S02, Se and CH2, and wherein R represents hydrogen or alkyl of 1 to 4 carbon atoms, with the proviso that at least one of the Xx is not CH: .. or a salt, prodrug and complex thereof, for the preparation of a pharmaceutical composition for the treatment and / or prevention of diabetes. In particular, the present invention relates to the use of a compound of the general formula I, wherein the diabetes is type I diabetes. A preferred embodiment of the present invention relates to the use of a compound of the general formula I, wherein Diabetes is type II diabetes. Additional modalities refer to types of diabetes selected from the group comprising secondary diabetes such as pancreatic diabetes, extravascular / endocrine or drug induced extirpation, or exceptional forms of diabetes such as diabetes 1 ipoat rófica, miatónica or diabetes caused by disturbance of insulin receptors.

One embodiment of the invention is the use of a compound of the formula I wherein m > 13. A currently preferred embodiment of the invention comprises formula I, wherein Xi = 3 is selected from the group consisting of 0, S, SO, S02 and Se, and wherein Xi = 5-25 is CH2. Tetradecylthioacetic acid (TTA) and selenoacetic t et radeci 1 acid (TSA), for example Xi = 3 is sulfur and selenium, respectively is one of the currently preferred compounds. In still another aspect of the present invention, this relates to the use of a compound of the formula I for the preparation of a pharmaceutical composition for the treatment and / or prevention of the syndrome mui t imet abó! ico called "Met abó licic syndrome" which among other things is characterized by hyperinsulinemia, insulin resistance, obesity, glucose intolerance, diabetes mellitus type 2, dyslipidemia and / or hypertension. An additional aspiration of the invention relates to a method for the treatment or prevention of a diabetic condition, said method comprising the step of administering to an animal in need thereof an effective amount of fatty acid analogs of the formula general (I): CH3- [CH2] T- [X1-CH2) n-COOR where n is an integer from 1 to 12; and where m is an integer from 0 to 23, and where i is an odd number indicating the position relative to COOR, and - where XL independently from each other are selected from the group comprising O, S, SO, S02, Se and CH2, and wherein R represents hydrogen or alkyl of 1 to 4 carbon atoms, - with the proviso that at least one of the Xx is not CH2. or a salt, prodrug or complex thereof. According to the method indicated above, preferred embodiments are as follows: said animal is a human, the animal is a farm animal, such as gallinaceous birds, bovine, ovine, caprine or porcine mammals.

The animal is a pet or pet, such as a dog or a cat. The treatment involves the administration to a patient in need of such treatment, of a therapeutically effective concentration that is maintained continuously in the blood of the animal for the duration of the period of its administration. In addition, the invention relates to a pharmaceutical composition for the prevention and / or treatment of a diabetic condition. Preferably, the pharmaceutical composition comprises in admixture with the fatty acid analogues, a pharmaceutically acceptable carrier or excipient. In addition, the invention relates to methods for the treatment and / or prevention of hyperglycemia, hyperinsulmia or reduced sensitivity to insulin, said method comprising the step of administering to an animal in need thereof, an effective amount of fatty acid analogs of the general formula (I): The invention also relates to a nutritional composition comprising an amount of fatty acid analogs of the general formula (I): effective to reduce, or to prevent an increase in the concentration of glucose in the blood of human or non-human animal. The invention also relates to the novel fatty acid analogues of the general formula I CH3- [CH2] m- [X1-CH2) n-COOR where n is an integer from 1 to 12; and - where I get an integer from 0 to 23, and where i is an odd number that indicates the position relative to COOR, and where Xx independently of one another are selected from the group comprising 0, S, SO, S02, Se and CH2, and wherein R represents hydrogen or alkyl of 1 to 4 carbon atoms, with the proviso that at least one of the Xx is not CH; > . or a salt, prodrug or complex thereof.

LEGENDS OF THE FIGURES Figure 1 shows the effect of TTA on weight gain for rats, administered with a high fat diet. Figure 2 shows the effect of TTA on weight gain for rats given a diet high in sucrose. Figure 3 shows that treatment with TTA prevents hyperinsulinemia induced by high fat content in the diet. Figure 4 shows that TTA treatment prevents insulin resistance induced by the high fat diet. Figure 5 shows that the treatment with TTA reduces blood insulin and glucose concentrations in Zucker (fa / fa) r 5-week-olds. Figure 6 shows that treatment with TTA reduces blood insulin and glucose concentrations in Zucker (fa / fa) rats at 4 months of age (Figure 5B). Figure 7 shows that treatment with TTA decreases the response of plasma insulin to glucose.

Figure 8 shows that TTA increases mitochondrial ß-oxidation.

ADMINISTRATION OF THE COMPOUNDS OF THE PRESENT INVENTION As a pharmaceutical medicament, the compounds of the present invention can be administered directly to the animal by any suitable technique, including parenterally, intimately, orally, or by absorption through the skin. These can be admired locally or systemically. The specific route of administration of each agent will depend, for example, on the medical history of the animal. Examples of parenteral administration include subcutaneous, intramuscular, intravenous, intial and intraperitoneal administration. As a general proposition, the total pharmaceutically effective amount of each of the compounds administered parenterally per dose will preferably be in the range of about 5 mg / kc / day to 1000 mg / kg / day of the patient's body weight, although, as previously noted, this will be subject to great care of therapeutic discretion. For TTA it is expected that a dose of 100-500 mg / kg / day is preferable, and for TSA the dose could probably be in the range of 10 to 100 mg / kg / day. If administered continuously, the compounds of the present invention are each typically administered by 1 to 4 injections per day or by continuous subcutaneous infusions, for example, using a mini-pump. An intravenous bag solution can also be used. The key factor in the selection of an appropriate dose is the result obtained, as measured by decreases in total body weight or the ratio of fat to lean mass, or by other criteria for measurement, control or prevention of Obesity, or prevention of conditions related to obesity, as they are considered appropriate by the practitioner. For parenteral administration, in one embodiment, the compounds of the present invention are generally formulated by mixing each to the desired degree of purity, in a unit dose injectable form (solution, suspension, or emulsion) with a pharmaceutically carrier. acceptable, for example, one that is non-toxic to patients at the doses in concentrations employed and that is compatible with other ingredients of the formulation. In general, the formulations are prepared by contacting the compounds of the present invention each uniformly and intimately with liquid carriers or finely divided solid carriers or both. Later, if necessary, the product is shaped into the desired solution. Preferably, the carrier is a parenteral carrier, more preferably a solution that is isctonic with the patient's blood. Examples of such carrier vehicles include water, saline, Ringer's solution, and dextrose solution. Non-aqueous vehicles such as fixed oils and ethyl oleate are also useful herein, as well as the 1 iposomes. The carrier may suitably contain minor amounts of additives such as substances that improve isotonicity and chemical stability. Such materials are non-toxic to patients at the doses and concentrations employed, and include buffers such as phosphate, citrate, succinate, acetic acid, and other organic acids or their salts; antioxidants such as ascorbic acid; immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamic acid, aspartic acid, or arginine; monosaccharides, disaccharides, and other carbohydrates including cellulose and its derivatives, glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; counterions such as sodium; and / or nonionic surfactants such as poly isorbate, poloxamers, or PEG. For oral pharmacological compositions, carrier materials such as for example water, gelatin, gums, lactose, starches, magnesium stearate, talc, oils, polyalkylene glycol, petroleum jelly and the like can be used. Such a pharmaceutical preparation may be in unit dosage form and may further contain other therapeutically valuable substances or conventional pharmaceutical adjuvants such as preservatives, stabilizing agents, emulsifiers, buffers and the like. The pharmaceutical preparations can be in conventional liquid forms such as tablets, capsules, dragees, ampoules and the like, in conventional dosage forms such as dry ampules, and as suppositories, and the like. The treatment with the present compounds may occur without, or may be imposed with, a restriction on the diet such as a limit on the daily food or on the ingestion of calories, as desired by the individual patient. In addition, the compounds of the present invention are suitably administered in combination with other treatments to combat or prevent obesity. The invention will be more fully understood by reference to the following examples. These should not, however, be considered as limiting the scope of the invention.

EXPERIMENTAL SECTION METHODS Obese rats Zucker (fa / fa). The obese Zucker rats (fa / fa) used in this study were reared at the U 465 INSERM animal facility from pairs originally provided by the Harriet G. Bird Laboratory (Stow, MA USA). Unless stated otherwise, the animals were kept under a constant light-dark cycle (light from 7:00 am to 7:00 pm) at 21 ± 1 ° C and were given free access to food and water. Three rats were housed per cage. The gains in weight were recorded daily.

Rats istar Male Wistar Charles River rats weighing 280-358 grams were purchased from AnLab Ltd.

(Prague, Czech Republic) and housed in wire mesh cages at a temperature of (22 ± 1 ° C) and controlled light (after 7:00 a.m. to 7:00 p.m.). These were given free access to food and water. Three rats were housed per cage. The gain in weight and the ingestion of food were recorded daily.

Diets (given in% by weight) used in feeding experiments Standard food diet: The rats were fed a rats feed ST1 for Standard Laboratory, Velaz, Prague, Czech Republic.

Diet with high sucrose content (HS) 50. 3% sucrose, 4.8% gelatin, 3.2% hay, 2.3% vitamins and minerals, 8.7% yeast, 8.7% lech powder, 12.3% casein, 9% meat tallow, 1% oil sunflower.

HS + TTA: the same as HS + 0.3% of TTA dissolved in beef tallow. HS + fish oil (FO): meat tallow and sunflower oil is replaced by 10% Triomar. Tr Lomar is from Pronova Biocare, Norway and contains 33.4% EPA, 3.1% DPA and 20.2% DHA.

High fat content (HF): 1.9% gelatin, 5.7% wheat bran, 7.7% vitamins and minerals, 25.4% corn starch, 25.7% casein, 26.8% meat tallow and 7.1% oil sunflower. HF + TTA: the same + 0.4% of TTA dissolved in beef tallow. HF + FO: 10% of meat tallow is replaced by 10% of Triomar.

Glucose tolerance tests, intravenous Male Zueker (fa / fa) rats (5 weeks old) were anesthetized after a 5 hour fast, by peritoneal injection of pentobarbi such sodium (50 mg / kg). The rats were injected. with glucose (0.55% g / kg) in the saphenous vein and blood samples were collected from the tail vein in hepar tubes initiated at the times of 0, 5, 10, 15, 20 and 30 minutes after loading of glucose. The samples were kept on ice, centrifuged and the plasma was stored at -20 ° C until analysis.

Euglycemic hyper insulic clamp After 21 days in their respective diets (see above), the rats were anesthetized by injection of xylazine hydrochloride (Rometar SPOFA, Prague, Czech Republic, 10 mg / ml) and ketamine hydrochloride (Narkamon SPOFA, Prague, Czech Republic; 75 mg / ml), and adjusted with cannulas for the chronic carotid artery and the jugular vein as described by Koopmans et al. (Koopmans SJ, .and collaborators Biochim Biophys Acta, 1115, 2130-2138 1992). The cannulated rats were allowed to recover two days after the surgery before the clamping studies that were carried out according to Kraegen and collaboracores (Kraegen E. W. et al., Am J Physiol, 248, E353-E362 1983). Thus, on the third day after surgery, the unrestricted conscious rats were administered with a continuous infusion of porcine insulin (ActrapLd, Novo Nordisk, Denmark) at a dose of 6.4 mIJ per kg per minute, to reach insulin levels Plasma in the upper physiological range. The arterial blood glucose concentration was confirmed at the basal fasting level, by variable infusion of a glucose solution at 30% w / v (Leciva, Prague, Czech Republic). Blood samples for the determination of plasma glucose and insulin concentrations were obtained every 15 minutes after the start of glucose infusion. After 90 minutes, the rats were disconnected from the infusions and immediately decapitated, the blood was collected for plasma separation, adipose tissue pads from the liver and the epididymis were dissected and weighed.

Measurement of plasma parameters Glucose concentrations (GLU, Boehringer Manheim, Germany), free fatty acids (NEFA, C ACS-ACOD equipment, Wako Chemicals, Dalton, USA) and b-hydroxybutyrate (equipment 310-A, Sigma Diagnostics Inc., St Louis, USA) were measured using enzymatic methods. Insulin concentrations were determined with radioimmunoassay using (CIS bio International, Gif sur Yvette, France) the use of rat insulin as standard in Zucker rats. In Wistar Charles River rats, plasma glucose concentrations were measured with the help of the Beckman glucose analyzer (Fullerton, CA, United States). Plasma insulin levels were measured using a RIA team from Lineo Research Inc. (St. Charles, MO, United States). The phospholipids were measured by the enzymatic method of bioMérieux, Marcy-1 'Et oi le, France, triacylglycerol by Technicon Method No. SA4-0324L90, United States and cholesterol by Technicon method No. SA4-0305L90 , U.S.

Preparation of nuclear and mitochondrial post-nuclear fractions and measurement of magnetic activity Livers freshly isolated from old individual Zucker rats were homogenized with ice-cooled sucrose buffer (0.25 M sucrose, 10 mM ipH 7.4 HEPES) and 2 mM EDTA). The postnuclear and mitochondrial fractions were prepared using preparative differential centrifugation according to DeDuve, C., and collaborators Biochem. J., 60, 604-617 1955). The modifications, purity and performance were as described at the beginning (Garras, A., and collaborators Biochim, Biophys, Acta, 1255, 154-160 1995). Acid-soluble products were measured in the post-nuclear and mitochondrial enriched fractions, using [l-14C] -palmitoyl-CoA and [1 -14C] -palmi t or i 1-L-carni t ina (Radiochemica 1 Center, Amersham, England) as substrates as described at the beginning (Willumsen, N., et al., J. Lipid Res., 34, 13-22 1993). The carnitine-palmit oiltrans-I and -II activities were measured in the postnuclear and mitochondrial fraction essentially as described by Bremer (Bremer, J., Biochim, Biophys, Acta, 665, 628-631, 1981), and 3-hydroxy-3-met i lglut ar i 1-CoA-s int a sa was measured according to Clinkenbeard and co-workers (Clinkenbeard, KD et al., J. Biol. Chem, 250, 3108-3116 1975) in the mitochondrial fractions.

RNA analysis RNA extraction (Chomc zyns ki, P., et al., Anal. Biochem., 162, 156-159 1987), analysis of Northern blotting and transfer or spotting of RNA on nylon filters, and hybridization to the immobilized RNA, they were performed as described at the beginning (Vaagenes, H., et al Biochem, Pharmacol., 56, 1571-1582 1998). The following cDNA fragments were used as probes: CPT-I (Esser, V. et al., J. Biol. Chem., 268-5817-5822 1993), CPT-II (Woeltje, KF et al., J. Biol. Chem., 265, 10720-10725 1990), 3-h? Drox? -3-metigltaryl-CoA-smtase (Ayte, J., et al., Proc. Nati, Acad. Sci. United States, 87, 3874- 3878 L990), and the hormone-sensitive lipase (Holm, c., Et al., Biochim, Biophys. Acta, 1006, 193-197 1989). Relative levels of RNA expression were estimated as the amounts of radioactive probe hybridized to the respective 28S rRNA levels.

RESULTS Example 1. Preparation and characterization of the compound a) Synthesis of the novel compounds Giassic acids with the heteroatom in variable positions were synthesized according to the generic description for the 3-substituted analogues (see below) with the following modification: Alkyl-Hal was replaced by Alkanoic-Hal and HS-CHCOOR was replaced by alkyl -SH. The following fatty acid analogues have been prepared and characterized: The purification of the products was as described below. Purity > 95% The structure was verified by mass spectrometry. b) The synthesis of the fatty acid analogues 3- their t i tidos The compounds used according to the present invention, wherein the substituent X1 = 3 is a sulfur atom or a selenium atom, can be prepared according to the following general procedure: X is a sulfur atom: The t io-sus t-tido compound used according to the present invention can be prepared by the general procedure indicated below: Base Alkyl-Hal + HS-CH2COOR === > Alk i 1 - S -CH 2 -COOR The compound with sulfur, namely tetradecylthioacetic acid (TTA), (CH 3 - (CH 2) - 3-S-CH 2 -COOH was prepared as shown in European patent EP-345,038 .

X is a selenium atom The selenium substituted compound used according to the present invention can be prepared by the following general procedure: 1. Alkyl-Hal + KSeCN = > Alkyl 1 -SecN ... 2. Alkyl-SeCN + BH4"=> Alkyl-Se" 3. Alkyl-Se "+ 02 => Alkite-1-Se-Se-Alki- This compound was purified by careful crystallization from ethanol or methanol.

BH4"4. Alkyl-Se-Se-Alkyl = > 2-alkyl-Se 5. Alkyl-Se" + Hal-CH2-COOH = > Alkyl-Se-CH2-COOH The final compound, for example, when the alkyl is tetradecyl (CH3- (CH2)? 3-Se-CH2-COOH (t-selenioacetic acid (TSA)) can be purified by crystallization from diethyl ether and hexane This product can be completely characterized by NMR, IR and molecular weight determination The method for the synthesis and isolation of these sulfur and selemo compounds, and the compound where X of the formula I is oxygen (0), sulfur oxide I (SO) and sulfur dioxide (S02) are described in European Patent No. 345,038, and International Patent Application No. WO 97/03663.

E] emplo 2 TTA toxicity study A 28-day toxicity study in dogs according to the GLP guidelines (good laboratory practices) has been carried out by Corning Hazleton (Europe), England. Oral administration of TTA at dose levels up to 500 mg / kg / day was generally well tolerated. Some parameters related to lipids were decreased in animals given high doses. This is consistent with the pharmacological activity of TTA.

The dose level of 500 mg / kg / day also caused the loss of body weight. There was no evidence of toxicity at dose levels of 50 or 500 mg / day / kg. The tests for the mutagenic activity have been carried out with Covance Laboratories Limited, England. It was concluded that TTA and TSA did not induce mutations in the strains of Salmonel typhimurium and Escherichia coli. In addition, TTA was not mutagenic as was tested in mouse lymphoma cells and L5178Y. The concentration of the tested compounds in S. typhymur ium and E. coli in 3-1000 mg / plate (TTA), 2-5000 mg / plate (TSA). In mouse lymphoma cells L5178Y, the concentration was 2.5-50 mg / ml. TSA and TTA were found to be non-mutagenic in these tests. TSA and TTA have been tested for chromosomal aberrations in cultured Chinese hamster ovary cells and were not induced to aberrations by the doses tested (12-140 mg / ml). The compounds of the present invention are therefore potentially useful as pharmaceutical compounds in this regard.

E j us 3 TTA induces a decrease effect in lipids in obese animals Obese male Zucker fa / fa rats, weighing 100 g at the beginning of the experiment, were housed in pairs in metal wire cages in a room kept at 12-hour light-dark cycles and a constant temperature of 20 ± 3 ° C. The animals were acclimated for at least one week under these conditions before the start of the experiment. The TTA (acid t et radeci 1 t ioacético) prepared according to the procedure described previously, and palmitic acid (control) were suspended in carboxymethylcellulose at 0.5% (w / v) (CMC). Six animals were used in both groups. TTA (tet radeci lt ioacetic acid) and palmitic acid were administered at a dose of 300 mg / day / kg body weight., by gastric intubation (fattening) once a day for 10 days. The rats were fasted for 2 hours before the termination of the experiment. Blood and organs were collected. The concentrations of lipid in the plasma were determined using a self-measuring device, as described in the methods section. The results obtained are reported in Table 1.

TABLE 1 Effect of TTA on lipid levels in obese Zucker fa / fa rats.

The results clearly show that TTA decreases the levels of triglycerides, cholesterol and phospholipid in the plasma.

E j us 4 TTA and TSA induce a lipid lowering effect in normal animals (Wistar rats) Male Wistar rats, weighing 180-200 g at the beginning of the experiment, were housed individually in metal wire cages in a room maintained at 12-hour light-dark cycles and at a constant temperature of 20 ± 3 ° C. The animals were acclimated for a week under these conditions before the start of the experiments. TTA, TSA and eicosapent aenoic acid (EPA) were suspended in 0.5% (w / v) carboxymethylcellulose (CMC). Six animals were used for each treatment, and a 0.5% solution of CMC was administered to rats as control. After administration of the test compound, the rats were fasted for 12 hours and anesthetized with haloet ano. The EPA and the fatty acid derivatives were administered by gastric intubation (priming) once a day for 7 days. Blood samples were collected by cardiac puncture, and lipid concentrations in the plasma were determined as described in the methods section. The results are given in table 2.

Table 2 Effect of TTA, TSA and EPA on plasma lipid levels in rats Table 2 shows that TTA exhibits a good blood lipid lowering effect in rats. It would appear that a 100-fold higher dose of EPA is necessary to obtain the same decrease in plasma lipid concentration as obtained for TSA. In addition, the substituted fatty acid compounds of the present invention are much more effective than pure EPA and fish oil in lowering plasma lipids. Therefore, these are potentially useful as medical compounds.

E j us 5. Influence of TTA on high-fat diets, fed Wistar rats Charles River Wistar Charles River male rats (280-360 g) were fed 3 different diets (see methods) for 3 weeks at d 1 ibi t um. After this, they were sacrificed by decapitation, the adipose tissue pads of the liver and the epididymis were dissected and weighed. By feeding the Wistar rats with the high-fat diet, the weight of the epididymal fat pad and the re-treper implant were increased. Treatment with TTA prevented the increase in tissue in adipose tissue mass and this effect was independent of food consumption, which was identical (high fat content: 15.1 ± 1.1 vs. high fat content + TTA: 14.8 ± 1.3 g / rat / day.

Table 3 Influence of high fat diets with and without TTA supplementation for three weeks on the gain in body weight, liver weight and adipose tissue weights in Charles River Wistar rats fed a high fat diet.

The data are given as the mean ± SEM E j us 6. TTA decreases the total body weight of the normal rats 2 groups of 6 male Wistar rats were randomly selected, and studied for weight development over a period of 12 weeks. The body weight of each Wistar rat was measured at the beginning of the experiment. All the animals in both groups individually received the same amount of food (nutrition) during the experimental period of 12 weeks. All the animals in one of the groups were orally administered with the medicine that comprises TTA. The other group was the control group (CMC). After the 12-week period, the body weight of the rats was measured again. The results are given in Table 4 and show that oral administration of TTA leads to significant loss in weight.

Table 4 Effect of TTA on the body weight of Wistar male rats after 12 weeks of treatment E j us 7. Influence of TTA on high-fat diets administered to Wistar Charles River rats Figure 1 shows the cumulative values for the gain in weight (g) / total food eaten (g) in 3 weeks. The values were calculated by taking the daily average weight gain and dividing it by the average amount of food eaten that day. See the section on methods for abbreviations and the specification of diets. The composition of the diets is given in the methods section.

Example 8 Influence of TTA on high sucrose content diets administered to Wistar Charles River rats Figure 2 shows the cumulative values for weight gain (g) / total food eaten (g) in 3 weeks. The values were calculated by taking the daily average weight gain and dividing it by the average amount of food eaten that day. See the section on methods for abbreviations and the specification of diets. The composition of the diets is given in the methods section.

E j psa 9. Influence of TTA on body weight gain, liver and adipose tissue weight in obese animals TTA was also tested for its effect on liver and adipose tissue weight. The results are indicated in table 5. Zucker obese male rats (fa / fa) of 5 weeks of age were fed TTA, 300 mg / kg / day suspended in 0.5% CMC. The control animals received only CMC. After 11 days of treatment, the rats were sacrificed by cervical dislocation, and the adipose tissue pads of the liver and epididymis were dissected and weighed. The data are the means ± SD of 6 animals of the control group and 6 animals in the experimental group.

Table 5 Influence of TTA on body weight gain, weight of liver and adipose tissue in obese young Zucker rats (fa / fa).

Example 10. TTA induces a reduction in weight in dogs 3 male dogs (4 to 6 months old) were housed alone during the days. Each animal was offered 400 g of SQC diet A every morning after dosing, and any residue from the diet was removed in the afternoon. The drug was administered orally in capsules once a day for 28 days.

Table 6. Mean body weights of male dogs treated with 500 mg / kg / day of TTA for 4 weeks.

E xemployment 11. Treatment with TTA prevents the induction of hyperinsulinemia by HF diet in normal rats Rats weighing 280-360 g were divided into 3 groups (n = 6) and fed three different diets: standard feed for rats, high fat diet (HF) and HF supplemented with TTA. After 21 days in their respective diets, blood was collected after an all-night fast, from the vein of the tail. The data is shown as the mean ± SEM. The results were analyzed by ANOVA (analysis of variance) and the different letters denote the statistical significance (p <0.05). Figure 3 shows that TTA treatment prevents hyperinsulinemia induced by the high-fat diet in Wistar Charles River rats.

Example 12 Treatment with TTA prevents insulin resistance induced by high-fat diet (HF) in normal rats Rats weighing 330 ± 20 g were divided into 3 groups (n = 9) and fed three different diets: standard feed for rats, high fat diet (HF) and HF supplemented with TTA. After 21 days of their respective diets, a 90 minute euglycemic hypermic retention was performed in unrestricted conscious animals as described in Materials and Methods. The rate of glucose infusion (GIR) was determined from the period of the clamp or clamp where the blood glucose remained stabilized, for example between 45-90 minutes after the start of fixation or clamping. The data are presented as the mean ± SEM. An euglycemic hyperinsulinic clamp or hyperinsulin protocol was established to test whether the highest TTA in the diet could improve the deterioration of insulin action, induced by feeding with high fat content, in rats. The 90-minute hyperinsemic ulcerative euglycemic clamp resulted in maximum plasma glucose and plasma insulin levels that were not different in the three groups studied. There was a significant reduction in the rate of exogenous glucose infusion (GIR) required to maintain euglycemia in the HF group (Figure 4) compared to Wistar rats fed the standard diet. Interestingly, TTA overfeeding of the HF diet prevented the development of insulin resistance in these rats as evidenced by completely normal GIR. This indicates a beneficial effect of TTA on the action of insulin in vivo. Figure 4 shows that treatment with TTA prevents insulin resistance induced by the high-fat diet in Wistar Charles River rats.

E xemplo 13 The effect of TTA on plasma levels of insulin and glucose in obese animals -week-old Zucker rats (fa / fa As shown in Figure 5, treatment with TTA reduced the blood insulin concentration by almost 40%, while the blood glucose concentration was reduced by approximately 15%. The rats were administered with TTA at a dose of 300 mg / kg / day suspended in 0.5% CMC (n = 6) by oral priming. After 11 days of treatment, the rats were sacrificed by cervical dislocation. The blood was collected and the insulin and glucose levels measured as indicated in the methods section. The data are the means ± S.D. According to Zucker, L.M. and collaborators (Sparks, J. D. et al., Metabolism, 47, 1315-1324 1998), these young animals have not developed hyperglycemia.

Obese rats Zucker (fa / fa) 4 months old Figure 6 shows the effect on TTA on insulin and blood glucose levels in Zucker (fa / fa) rats at 4 months of age, for example rats that have developed hyperglycemia (Sparks, JD et al., Metabolism, 47, 1315- 1324 1998). The rats were administered with a standard diet, either with (n = 5) or without (n = 6) 0.15% TTA. After 21 di. As of treatment, blood was collected and insulin and glucose levels were measured. The data are the means ± S.D.

E xemployment 15. Treatment with TTA decreases the response of plasma insulin to glucose To investigate whether treatment with TTA resulted in or not an improvement in the action of insulin on glucose utilization, an intravenous glucose tolerance test (IVGTT) was performed. In Zucker rats at 5 weeks of age (fa / fa), treatment with TTA resulted in a significantly lower plasma insulin response towards glucose (Figure 7A). The IVGTT glucose curves were normal and comparable between the control and TTA treated rats (Figure 7B).

E xemplo 16 The effect of TTA on mitochondrial ß-oxidation Obese Zucker (fa / fa) rats were administered with a standard feed either with (n = 6) or without (n = 5) 0.15% TTA. After 21 days of treatment, the rats were sacrificed by cervical dislocation and the livers were removed. The mitochondrial fractions were isolated from the individual livers. Oxidation rates of fatty acids were measured using [1 -14 C] -palmi t oi 1 -CoA or [1-14C] -pa lmi t oi 1 -L-carni t ina as substrate (A) CPT-I ( B) and CPT-II (C) were measured in the mitochondrial fractions. RNA purification and hybridization experiments were also performed. The relative levels of mRNA were determined by densitometry scanning of the autoradiograms and the different mRNA levels were normalized to the 28S rRNA and the means for the controls were adjusted up to 1. The formation of the acid-soluble products in the Obese control animals were 1.3 ± 0.7 and 5.3 ± 2.2 nmol / g liver / minute using pa lmi t oi 1 -CoA and palmi toi 1 -L-carnitine as substrates, respectively. The CPT-I activity in control rats was 22.14.9 nmol / g liver / minute, and the CPT-II activity in the control rats was 270 + 115 nmol / g liver / minute. The values are expressed as the mean i S.D. The administration of TTA increased the plasma concentrations of ketone bodies, resulting in a marked decrease in the body ratio FFA / ketone (Table 7). These data indicate that treatment with TTA of obese 4-month-old Zucker (fa / fa) rats increased hepatic mitochondrial ß-oxidation and ketogenesis. Of course, treatment with TTA of obese Zucker (fa / fa) rats increased hepatic fatty acid oxidation more than 7 times as measured with palmi toi 1-CoA and palmi toyl-L-carnitine as substrates (Figure 8A). This induction of β-oxidation was accompanied by an increase in the activity of mRNA levels of CPT-I (Figure 8B) and CPT-II (Figure 8C). In addition, the activities of speed-limiting enzymes in ketogenesis were increased (Table 7).

TABLE 7 Influence of TTA on the plasma concentration of free fatty acids (FFA) and cetomach bodies (4-hydroxybutyrate) in obese old Zucker rats.

The data are the means ± S.D. of six animals in the control group and in the experimental group. The free fatty acids (FFA) and the cetomach bodies (4-hydroxybut i rat o) were measured in plasma and the activities of 3- h? Drox? -3-met? Lglutar? L- (HMG) -CoA- were measured. Smtasa in fractions my tocondría prepared from the liver of rats Zucker (fa / fa) obese, male, 21 weeks old, admin Lst radas either with a standard diet control or a standard diet enriched with 0.15% TTA for 15 days.

E xemplo 17 The effect of TTA on hepatic levels of triacylglycerol The oxidation of mitochondrial fatty acids significantly increased, caused by TTA will reduce the availability of fatty acids for esterification. The synthesis of glycerol and cholesterol is thus reduced, and the secretion of VLDL from the liver is diminished. This is reflected at a decreased level of blood glucose in the liver, reduced plasma glucose, and reduced adipose tissue mass. The basal and total lipolysis are not changed (data not shown) and the ratio between plasma free fatty acids and ketone bodies is diminished (data not shown). This indicates an increased flow of fatty acids from the peripheral tissues to the liver for oxidation.

Even a hepatic level of triacylglycerol can be alleviated by TTA. By feeding the rats with an inhibitor of fatty acid oxidation, the level of hepatic thyroid is increased resulting in fatty liver. Tet radeci 1 - 4-t ia-propiónico acid (TTP) is a fatty acid analogue with a sulfur atom in position 4. This analogue inhibits the ß-oxidation of fatty acids due to the formation of a mitochondrial inhibitor . Feeding the rats with this analogue results in the formation of fat. However, if rats are fed a combination of TTA and TTP, fatty liver formation is avoided (Table 8). This provides evidence that TTA can be used for the treatment of conditions with an increased hepatic level of metabolism. Male Wistar rats had free access to water and food for rat maintenance. These were fed with palmitic acid or fatty acid analogues suspended at 0.5% CMC for 6 days. In some experiments TTA or TTP were fed for 3 days before feeding both for 6 days. At the end of the experiment, the rats were fasted overnight, sacrificed, the liver was removed and homogenized. Triacylglycerol was measured in the homogenate.

Table 8 Hepatic levels of t riacilgl icerol in rats treated with palmitic acid and fatty acid analogues for 6 days (TTA: 150 mg / kg / day - TTP: 300 mg / kg / day).

Use 1 The fatty acid analogs have been synthesized where the sulfur atom is moved to the posterior positions of the carboxyl group of the fatty acid. When the sulfur atom is placed in the positions on the carbon chain with odd numbers (5, 7, 9, etc.), these analogs will be partially β-oxidized. The β-oxidation removes the two carbon atoms at a time from the carboxy terminus of the fatty acid, and such analogs can thus be β-oxidized until the sulfur atom is in the 3-position. In this way, it is conceivable that such analogs may have biological effects similar to TTA. Experiments have shown that the fatty acid analogues related to having a sulfur atom in an odd numbered position on the carbon chain will all increase the mitochondrial ß-oxidation (Table 9). The mitochondrial ß-oxidation is measured as in example 16 with the use of [1-14C] -pa lmit oi 1 -L-carnitine as substrates.

Table 9 Effect of different fatty acid analogues on mitochondrial ß-oxidation in rat liver E xemployment 19 Fat obese, male Zucker rats, weighing 100 g at the beginning of the experiment, were housed in pairs in metal wire cages in a room maintained at 12 hours lu-cycles and a constant temperature of 2013 ° C. The animals were acclimated for at least one week under these conditions before the start of the experiment. TTA and palmitic acid (control), was suspended in 0.5% (w / v) of carboxymethyl Ice lulose (CMC) and administered at a dose of 300 mg / day / kg of body weight, by gastric intubation (fattening) once a day for 10 days. The rats were fasted for 2 hours before the end of the experiment. Blood and organs were collected. The total lipids were extracted from the liver and plasma. The lipids were evaporated, saponified and esterified before separation using a Cario Erba 2900 gas chromatograph.

Table 10 Effect of compound I (tet radecytic acid on the composition of fatty acids in obese Zucker fa / fa rats.

Table 10 shows that oral administration of TTA increases the level of oleic acid in liver and in plasma. Also, a delta-9-desaturated TTA product accumulated in the plasma and in the liver.

Claims (21)

1. The use of fatty acid analogs of the general formula (I): CH3- [CH2] m- [X1-CH2] n-COOR where n is an integer from 1 to 12; and where m is an integer from 0 to 23, and - where i is an odd number indicating the position relative to COOR, and where Xx independently of one another is selected from the group comprising O, S, SO, S02, Se and CH2, and - wherein R represents hydrogen or alkyl of 1 to 4 carbon atoms, with the proviso that at least one of the Xx is not CH2, or a salt, prodrug or complex thereof, for the preparation of a pharmaceutical composition for the treatment and / or prevention of diabetes.

2. The use according to claim 1, wherein the diabetes is type I diabetes.

3. The use according to claim 1, wherein the diabetes is type II diabetes.

4. The use according to claim 1, wherein the diabetes is a form selected from the group comprising secondary diabetes such as pancreatic, extrapancreatic / endocrine or drug induced diabetes, or exceptional forms of diabetes such as diabetes 1 ipoat ró f ica, miatónica or diabetes caused by the disturbance of insulin receptors.

5. The use according to claim 1, wherein m > 13 H.H.

The use according to claim 1, wherein X? = 3 is selected from the group consisting of O, S, SO, S02 and Se, and wherein X? = 5-25 SS CH2.

7. The use according to claim 6, wherein X1 = 3 is sulfur.

8. The use according to claim 6, wherein Xi = 3 is selenium.

9. The use of the fatty acid analogs of the general formula (I): CH3- [CH2] m- [Xi-CH2] n-COOR where n is an integer from 1 to 12; and - where m is an integer from 0 to 23, and where i is an odd number indicating the position relative to COOR, and where Xi independently of one another is selected from the group comprising O, S, SO, S02, Se and CH2, and wherein R represents hydrogen or alkyl of 1 to 4 carbon atoms, with the proviso that at least one of the Xi is not CH2, or a salt, prodrug or complex thereof, for the preparation of a pharmaceutical composition for the treatment and / or prevention of the metabolic syndrome called "metabolic syndrome" which is inter alia characterized by hyperinsulinemia, insulin resistance, obesity, glucose mtoLeranci, diabetes mellitus type 2, dyslipidemia and / or hypertension.

10. The use according to claim 9, wherein m > 13

11. The use according to the rei indication 9, wherein Xi = 3 is selected from the group consisting of O, S, SO, S02 and Se, and where X? = 5-25 is CH2.

12. The use according to claim 11, wherein X1 = 3 is sulfur.

13. The use according to claim 11, wherein Xi = 3 is selemo.

14. A pharmaceutical composition for the prevention and / or treatment of a diabetic condition in animals, the pharmaceutical composition comprises fatty acid analogues of the general formula (I): CH3- [CH2] m- [X1-CH2) n-C00R - where n is an integer from 1 to 12, and - where m is an integer from 0 to 23, and - where i is an odd number that indicates the position relative to COOR, and - wherein X independently from each other are selected from the group comprising O, S, SO, S02, Se and CH2, and - wherein R represents hydrogen or alkyl of 1 to 4 carbon atoms , - with the condition that at least one of the X-. is not CH, or a salt, prodrug or complex thereof.

15. A pharmaceutical composition according to claim 14, wherein the pharmaceutical composition comprises a mixture with the fatty acid analogs and a pharmaceutically acceptable carrier or excipient.

16. The use according to claim 14, wherein m > 13

17. A pharmaceutical composition according to rei indication 14, wherein X1 = 3 is selected from the group consisting of 0, S, SO, S02 and Se, and wherein X1 = 5-25 is CH2.

18. A pharmaceutical composition according to the rei indication 17, wherein X x = 3 is sulfur.

19. A pharmaceutical composition according to claim 17, wherein X1 = is selenium.

20. A nutritional composition comprising a quantity of fatty acid analogs of the general formula (I): CH3- [CH2] m- [X1-CH: -COOR - where n is an integer from 1 to 12, and - where m is an integer from 0 to 23, and - where i is an odd number that indicates the position relative to COOR, and - where Xx is independently one of the another is selected from the group comprising 0, S, SO, S02, Se and CH2, and wherein R represents hydrogen or alkyl of 1 to 4 carbon atoms, - with the proviso that at least one of the X is not CH;:, or a salt, prodrug or complex thereof, effective to reduce, or to prevent an increase in the concentration of glucose in the blood of a human or non-human animal.

21. A novel fatty acid analogue of the general formula I CH3- [CH2] m- [Xi-CH2) n-COOR - where n is an integer from 1 to 12, and - where m is an integer from 0 to 23, and - where i is an odd number that indicates the position relative to COOR, and - where Xi is independently one of the another is selected from the group comprising 0, S, SO, S02, Se and CH2, and - wherein R represents hydrogen or alkyl of 1 to 4 carbon atoms, - with the proviso that at least one of the X -. Do not be CH2. or a salt, prodrug or complex thereof.