MXPA99008765A - Methods and compositions for treating and/or preventing non-insulin dependent diabetes mellitus (niddm) using specific retinoid compounds - Google Patents

Abstract

Methods for treating and/or preventing non-insulin dependent diabetes mellitus (NIDDM) in subjects having or at substantial risk of developing NIDDM, using specific retinoid compounds that are structurally related to 9-cis retinoid acid which induce the differentiation of preadipocytes into adipocytes, are provided. These compounds may be administered alone or in combination with other anti-diabetogenic agents such as thiazolidinediones.

Description

MOLECULES RELATED TO THE RETINOID FOR THE TREATMENT OF DIABETES MELLITUS NOT DEPENDENT OF THE INSULIN TECHNICAL FIELD OF THE INVENTION The present invention relates to the discovery that certain retinoid-related compounds that are structurally related to 9-cis-retinoic acid effectively induce the differentiation of preadipocytes to adipocytes. These compounds, and compositions containing them, are useful for the treatment and / or prevention of non-insulin-dependent diabetes mellitus (NIDDM).

BACKGROUND OF THE INVENTION Approximately 3 to 100 people suffer from diabetes. Of these people, a large proportion of them, approximately 90 to 95%, of the approximately 6 million people diagnosed with diabetes in the United States comprise non-insulin-dependent diabetes mellitus (NIDDM) or type II diabetes.

? F .: B5U5 NIDDM is characterized by insulin resistance and abnormalities related to the secretion and action of insulin. More specifically, NIDDM is characterized by hyperglycemia, the result of insulin resistance in peripheral tissue (skeletal muscle and adipose tissue), which occurs because the uptake / utilization of glucose stimulated by insulin is mitigated here; and in the liver, which occurs because insulin suppression of glucose output is insufficient. These deteriorations in the action of insulin play an important role in the development of elevated blood glucose in fasting and glucose intolerance. Currently, methods for the treatment of NIDDM and the prevention of NIDDM include diet and exercise (since NIDDM and insulin resistance correlate strongly with obesity). Also, oral administration of hypoglycemic drugs to control blood glucose levels is another means of treating NIDDDM. Such hypoglycemic agents include formations containing insulin and sulfonylurea. However, these therapies suffer from significant disadvantages, in particular, the appearance of potentially life-threatening hypoglycaemia, which is attributable to hyperinsulinemia. This is problematic since hypoglycemia is associated with an elevated risk of cardiovascular disease, the biggest killer of diabetics. Therefore, the provision of a method for the treatment of diabetes that does not increase the circulating insulin concentrations, could be highly beneficial. Recently, a new class of synthetic drugs, the thiazolidinediones (TZDs), has been identified, whose drugs have been shown to increase insulin sensitivity in patients who are resistant to this hormone. Thiazolidinediones apparently improve insulin resistance and normalize glucose and plasma insulin (where they are elevated) without causing a hypoglycaemic state, even when very high doses are administered. The insulin sensitizers of TZD, for example, ciglitazone, englitazone, pioglitazone, BRL 49653 (5 - [[4- [2- (methyl-2-pyridinylamino) -ethoxy] phenyl] met il] -2,4-thiazolidinedione) and troglitazone, improve the suppression of the output of hepatic glucose measured by insulin, and the uptake and utilization of glucose stimulated by insulin, by adipose tissue. Also, it has been reported that TZDs alter the expression of the glucose transporter (for example Glut 4) which contributes to the increased responsiveness to insulin. A specific member of TZD, troglitazone, was recently reported as effective against NIDDM in phase III clinical trials (Nolan et al, N. Engl. J. Med., 331: 1188-1193, 1994). The potency of TZDs as effective antidiabetic agents closely resembles the adipogenic action, for example, their ability to differentiate preadipocytes into adipocytes (Harris and Kletzien, Mol.Pharma col., 45: 439-445, 1994).; Wilson et al J. Med. Chem. , 39: 665-668, 1996). Also, it has been reported that TZDs convert myogenic cells into adipocyte-like cells (Teboul et al, J. Biol. Chem., 270: 28183-28187, 1995). At the molecular level the TZDs have shown that they function as regulators of the nuclear PPAR receptor? (Lehmann et al J. Biol. Chem., 270: 12953-12956, 1995) which is known to play an important role in adipogenesis (Spiegelman and Flier, Cel l, 87: 377-389, 1996; Tontonoz et al. Spiegelman, Cell, 79: 1147-1156, 1994). However, the exact cellular mechanism by which TZDs increase insulin sensitivity in the NIDDM is not clearly understood. In addition, a major problem with TZDs is their toxic side effects in vi vo. Very recently, PCT application WO 97/10819 (published on March 27, 1997, after the priority date of this application) described that certain types of retinoids, for example, RXR agonists when administered alone or in combination with PPAR agonists? such as thiazolidinedione compounds, could be used to treat NIDDM. However, specific retinoids of the present invention are not mentioned herein.

BRIEF DESCRIPTION OF THE INVENTION The present invention relates to the discovery that specific retinoid-related molecules, which do not show typical retinoid activities, can be used as therapeutic agents. These compounds, in contrast to normal retinoids, show reduced ability or no ability to induce differentiation of F9 teratocarcinoma cells or P12 pluripotent teratocarcinoma cells. However, these compounds potentially potentiate the differentiation of preadipocytes to adipocytes. This property, coupled with the fact that these molecules are well tolerated, for example, does not show the typical retinoid toxicities, makes them very suitable for use in the treatment and / or prevention of NIDDM.

BRIEF DESCRIPTION OF THE FIGURES Figure 1 compares the ability of various retinoids according to the invention to induce the differentiation of preadipocytes to adipocytes at different concentrations.

Figure 2 compares the ability of various retinoids according to the invention to induce the differentiation of preadipocytes to adipocytes in a period of seven days.

OBJECTIVES OF THE INVENTION An object of the invention is to provide novel and improved methods for the treatment and / or prevention of NIDDM. A more specific objective of the invention is to use the retinoid-related molecules, specific, structurally related to 9-cis-retinoic acid, which induce the differentiation of preadipocytes to adipocytes, for the treatment and / or prevention of NIDDM. Another specific objective of the invention is to provide the novel compositions adopted for the treatment or prevention of NIDDM comprising the combination of at least one molecule related to retinoid, structurally related to 9-cis-retinoic acid, whose molecule induces the differentiation of preadipocytes to adipocytes and at least one thiazolidinedione compound (TZD).

DETAILED DESCRIPTION OF THE INVENTION The present invention is based on the discovery that certain retinoid-like molecules, which do not show typical retinoid activities, show the ability to induce the differentiation of preadipocytes to adipocytes. Also, these molecules show no adverse side effects in vi vo. This adipogenic action makes these compounds, and the isomers or pharmaceutically acceptable salts thereof, very suitable for the treatment or prevention of NIDDM. In particular, these compounds must show an effective anti-diabetogenic action based on their adipogenic activity, similar to the TZDs. However, contrary to TZDs, these retinoid compounds should not show toxic side effects after intravenous administration, because these molecules do not show the typical retinoid toxicities and have been shown to be very well tolerated in animals. The retinoid-like molecules reported herein are described in the North American application No. 08/429, 0096, filed April 26, 1995, now allowed. This application also describes the methods for the synthesis thereof. More specifically, it has surprisingly been found that the following retinoid compounds effectively induce the differentiation of preadipocytes to adipocytes: 6- (3, 5, 5, 8, 8-pentamethyl-5, 6, 7, 8-tetrahydro-2-naphthyl) nicotinic acid (referred to in Examples as Cl) (Compound 1); 4- (3-ethyl-5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-2-naphthyloxy) benzoic acid (referred to in Examples as C2) (Compound 2); 4- (3, 5, 5, 8, 8-pentamethyl-5, 6, 7, 8-tetrahydro-2-naphthio) enzoic acid (referred to in Examples as C3) (Compound 3); 4- (3-isopropyl-5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-2-naphthyloxy) benzoic acid (Compound 4); 4- (5,6,7,8-tetra-idro-5,5,8,8-tetramethyl-2-naphthyl) benzoic acid (Compound 5); 4- (3, 5, 5, 8, 8-pentamethyl-5, 6,7,8-tetrahydro-2-naphthyloxy) benzoic acid (Compound 6); 4- (3-bromo-5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyloxy) benzoic acid (Compound 7); 3-methy1-4- (5, 6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthylthio) benzoic acid (Compound 8); Y 3-Meth1-4- (3-ethyl-5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyloxy) benzoic acid (Compound 9).

The structures for the retinoid-related molecules, previously identified as Compounds 1-9, are described on the following page.

As discussed above, and as shown in the following examples, these compounds effectively promote adipogenesis. Therefore, these compounds or the isomers or pharmaceutically acceptable salts thereof can be used for the treatment and / or prevention of NIDDM. In the use of the present compounds to treat and / or prevent NIDDM, a therapeutic / prophylactic composition comprising a therapeutically or prophylactically effective amount of at least one compound according to the invention, will be administered to a subject having or being in risk of developing NIDDM. Such pharmaceutical / therapeutic compositions will comprise a carrier, carrier or diluent which is pharmaceutically acceptable and compatible with the mode of administration regimen selected for the given composition, and a therapeutically or prophylactically effective amount of at least one compound according to the invention, or a pharmaceutically acceptable salt or isomer thereof. The administration of the compounds according to the invention can be carried out by any suitable means, for example, systemically, enterally, parenterally, topically or ocularly. However, oral administration is generally preferred. For enteral administration, the medicinal / pharmaceutical compositions may be in the form of tablets, gelatin capsules, sugar-coated tablets, syrups, suspensions, elixirs, solutions, powders, granules, emulsions, microspheres or nanospheres or lipid or polymeric vesicles that they allow a controlled release. For parenteral administration, the compositions may be in the form of solutions or suspensions for perfusion or injection. The compounds according to the invention are generally administered at a daily dose of about 0.1 mg / kg to 100 mg / kg of body weight, which are administered at a rate of 1 to 3 doses per day. For topical administration, pharmaceutical compositions based on the compounds according to the invention can be provided in the form of ointments, creams, milks, ointments, powders, balms, impregnated pads, solutions, gels, sprays, lotions or suspensions. These can also be provided in the form of microspheres or nanospheres or lipid or polymeric vesicles or polymeric patches and hydrogels that allow controlled release. These compositions for topical administration may, in addition, be provided either in anhydrous form or in an aqueous form. For ocular administration, these are mainly ocular washes. These compositions for topical or ocular application contain at least one compound according to the invention or one of its salts, at a concentration preferably in the range of 0.001% to 5% by weight relative to the total weight of the composition. The medical compositions according to the invention can, moreover, contain inert or far-amine-active additives. In particular, the compositions according to the invention may comprise other drugs that are suitable for the treatment or prevention of NIDDM. In a preferred embodiment, the therapeutic / prophylactic compositions of the invention will comprise at least one retinoid compound according to the invention, in combination with at least one thiazolidinedione compound such as ciglitazone., englitazone, pioglitazone, BRL 49653 (5 - [[4- [2- (methyl-2-pyridylamino) ethoxy] phenyl] methyl] 2,4-thiazolidinedione, and troglitazone The combination of these adipogenic agents should provide at least additive and potentially synergistic effects on adipogenic activity Also, the therapeutic or prophylactic compositions of the present invention may further comprise other drugs useful for the treatment or prevention of diabetes.The retinoid compounds of the present invention, as noted, will be preferably used to treat people already diagnosed with NIDDM, for example those who show an active disease condition, however, another important application of the present invention comprises the use of the retinoid compounds present for the prevention of UIDDM in people who are at risk substantial risk of developing diabetes, for example, due to genetic factors and / or other risk factors. Risk factors include, by way of example, obesity, and pancreas transplantation. Also, these compounds or the pharmaceutically acceptable salts thereof can be used for the treatment of persons who show the early signs, for example preclinical, of NIDDM. Methods for identifying people who show the early signs of NIDDM or those who are at substantial risk of developing NIDDM are known in the diabetic technique, and include the measurement of glucose levels. In order to further illustrate the present invention and the advantages thereof, the following specific examples are given, it being understood that they are intended to be illustrative and in no way limiting.

EXAMPLE I Effects of the Present Compounds at Different Concentrations on Adipogenesis Activity 3T3-L1 preadipocyte cells (ATCC) were selected at 5 × 10 4 cells per well in DMEM and 10% calf serum in 24-well culture plates. Two days after reaching the confluence, differentiation was induced by the addition of different compounds according to the invention, as well as the appropriate control compounds in DMEM containing 10% fetal calf serum. The media and compounds were changed every three days. The cells were fixed after 7 days post-confluence and the accumulation of lipid droplets in the cytoplasm was determined by staining with red oil 0 (Oil Red O). All cells including control (vehicle) cells were treated with the same volume of dimethyl sulfoxide (DMSO), at a level not exceeding 0.2% of the final concentration of the solvent. All-trans RA (tRA) and 9-cis-RA (9cRA) were used as the control compounds.

Staining with Red O Oil: Seven days after confluence, the cells were washed twice with PBS, fixed with 10% formalin and washed once more with PBS. The cells were then stained with 60% solution of Red O Oil (Oil Red O) for 30 minutes. The cells were then washed twice with water for 15 minutes each.

The Red O oil reserve solution was prepared from 0.5 g of Red O Oil dissolved in 100 ml of isopropanol, and this was filtered before dilution in PBS to make the 60% Red O Oil solution.

The results of these experiments are contained in Figure 1. These results establish that the compounds of the inventors induced the differentiation of preadipocytes to adipocytes. In contrast, the control compounds did not show similar activity. In addition, the results indicate that such differentiation was induced in a concentration-dependent manner.

EXAMPLE II Effects of the Present Compounds on Adipogenesis with Time The 3T3-L1 preadipocyte cells (ATCC) were seeded at 5 × 10 04 cells per well in DMEM and 10% calf serum in 24-well tissue culture plates. Two days after reaching the confluence, the differentiation was induced by the addition of the different compounds according to the invention in DMEM containing 10% fetal calf serum (Day 0). The media and compounds were changed every three days. The cells were fixed on the indicated days and processed as explained in Example I.

All cells, including control (vehicle) cells, were treated with the same volume of dimethyl sulfoxide (DMSO) never exceeding 0.2% of the final concentration of the solvent. As a positive control for differentiation, the cells were treated with 10 μg insulin per ml and 1 μM dexamethasone (Ins / Dex). The all-trans (tRA) lμM and 9-cis-RA (9cRA) 2 μM RA were used as additional controls. Cl, C2 and C3 were used at a final concentration of 1 μM. The results of these experiments are contained in Figure 2. These results show that the compounds according to the invention induced the differentiation of preadipocytes to adipocytes in a time-dependent manner. In contrast, control compounds, insulin, dexamethasone, all-trans RA and 9-cis-RA did not show similar activity. Therefore, the results obtained support that the compounds of the invention effectively induce adipogenesis in a dependent, time and concentration manner. Accordingly, they should provide effective therapeutic and / or prophylactic agents for the treatment and / or prevention of NIDDM in subjects in need of such treatment.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (15)

RE IVINDICATIONS Having described the invention as above, the content of the following claims is claimed as property:

1. A method for the treatment and / or prevention of non-insulin dependent diabetes mellitus (NIDDM) in a subject having or at risk of developing NIDDM, characterized in that the method comprises administering a prophylactically or therapeutically effective amount of minus one compound selected from the group consisting of: 6- (3, 5, 5, 8, 8-pentamethyl-5, 6, 7, 8-tetrahydro-2-naphthylthio) nicotinic acid; 4- (3-ethyl-5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-2-naphthyloxy) benzoic acid; 4- (3, 5, 5, 8, 8-pentamethyl-5, 6, 7, 8-tetrahydro-2-naphthylthio) benzoic acid; 4- (3-isopropyl-5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-2-naphthyloxy) benzoic acid; 4- (5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthylthio) benzoic acid; 4- (3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydro-2-naphthyloxy) benzoic acid; 4- (3-Bromo-5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyloxy) benzoic acid; 3-methyl-4- (5, 6, 7, 8-tetrahydro-5, 5, 8, 8-tetramethyl-2-naphthyl) benzoic acid; 3-me i1-4- (3-ethyl-5, 6, 7, 8-tetrahydro-5, 5,8,8-tetramethyl-2-naphthyloxy) benzoic acid; or a pharmaceutically acceptable salt thereof.

2. The method according to claim 1, characterized in that the method further comprises administering at least one other anti-diabetogenic agent.

3. The method according to claim 2, characterized in that the other diabetogenic agent is a thiazolidinedione.

4. The method according to claim 3, characterized in that the compound is selected from the group consisting of ciglitazone, englitazone, pioglitazone, (5- [[4- [2- (methyl-2-pyridiniumlamino) ethoxy] phenyl] methyl] -2, 4-thiazolidinedione) and troglitazone.

5. The method according to claim 1, characterized in that the compound is administered systemically, enterally, parenterally, topically or ocularly.

6. The method according to claim 1, characterized in that the compound is administered orally.

7. The method according to claim 1, characterized in that the compound is contained in a pharmaceutically acceptable form selected from the group consisting of tablets, gelatin capsules, sugar coated tablets, syrups, elixirs, solutions, powders, granules, emulsions, microspheres , nanospheres, lipid or polymeric vesicles that provide controlled release, ointments, creams, milks, ointments, powders, balsams, impregnated pads, gels, sprays, lotions and suspensions.

8. The method according to claim 1, characterized in that the compound is administered daily at a dose of about 0.01 mg / kg to 100 mg / kg of body weight, at a ratio of about 1 to 3 doses per day.

9. The method according to claim 1, characterized in that it is administered to a subject at substantial risk of developing NIDDM due to genetic factors and / or other risk factors.

10. The method according to claim 9, characterized in that the subject is a person at risk of developing type II recurrent diabetes.

11. The method according to claim 1, characterized in that the compound is administered by injection.

12. A composition adopted for the treatment and / or prevention of non-insulin dependent diabetes mellitus (NIDDM), characterized in that it comprises the combination of: (i) at least one compound related to the retinoid selected from the group consisting of: 6- (3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydro-2-naphthylthio) nicotinic acid; 4- (3-ethyl-5,5,8,8-tetramethyl-5,6,7,8-tetra idro-2-naphthyloxy) benzoic acid; 4- (3, 5, 5, 8, 8-pentamethyl-5, 6, 7, 8-tetrahydro-2-naphthylthio) benzoic acid; 4- (3-isopropyl-5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-2-naphthyloxy) benzoic acid; 4- (5, 6, 7, 8-tetrahydro-5, 5, 8, 8-tetramethyl-2-naphthylthio) benzoic acid; 4- (3, 5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-2-naphthyloxy) benzoic acid; 4- (3-bromo-5, 6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyloxy) benzoic acid; 3-methi 1-4- (5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthylthio) benzoic acid; 3-methi 1-4- (3-ethyl-5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyloxy) benzoic acid; or a pharmaceutically acceptable salt thereof; (ii) at least one thiazolidinedione compound; and (iii) a pharmaceutically acceptable carrier or excipient.

13. The composition according to claim 12, characterized in that the thiazolidinedione compound is selected from the group consisting of ciglitazone, englitazone, pioglitazone, (5- [[4- [2- (methyl-2-pyridinylamino) ethoxy] phenyl] methyl ] -2,4-thiazolidinedione) and troglitazone.

14. The composition according to claim 12, characterized in that it is adopted for oral administration.

15. The composition according to claim 14, characterized in that it is in a form selected from the group consisting of tablets, gelatin capsules, sugar coated tablets, syrups, elixirs, solutions, powders, granules, emulsions, microspheres, nanospheres, lipid vesicles or polymeric that provide controlled release, ointments, creams, milks, ointments, powders, balsams, impregnated pads, gels, sprays, lotions and suspensions.