RS51543B - NEW BICYCLIC HORMON SENSITIVE LIPase INHIBITORS - Google Patents

Abstract

Benzotriazoles of general formula I, characterized in that they denote: R1 to R8H, wherein one of these residues R2 or R3 can stand in place for: Br, Cl, CH3, CN, NH2, NO2, CF3, OCH3, phenoxy. benzoyl, CH (OH) -phenyl, S-cyclohexyl, CO-OCH3, or two substituents of this series are: R1 = C1 and R3 = CF3 or R2 = F and R3 = Cl; n is an integer 0, 1 or 2; any of the substituents R6 or R7 may stand in place of: R6 CH3, R7 CH3, C2H5; CH (CH3) 2, C (CH3) 3. CF3. Br, Cl, benzoyl or CO-OC2H5; or R6 and R7 are both CH3; or the ring instead of R6 and R7 may contain one double bond or R5 and R6 or R6 and R7 together with the C-atoms on which they are located stand in place for one ring attached to benzene or if n = 0, it can stand in place for cyclohexanedyl, wherein in the case of ring closure R6 / R7 this substituent may optionally be substituted once with NH2 or NO2 or once or twice with OCH3; iR7 and R8 together are cyclopentyl, diazirine or = CH2, with the exception of compounds R1 to R5 and R8 = H, n = 1 and R6 / R7 = fused to benzene and R1, R3 -R8 = H, R2 = CH3 and = 1. The application contains 11 more patent claims.

Description

Benzotriazoles are already known in various fields, for example in photochemistry (US 4,255,510, Kodak) or as orexin antagonists (WO 02/090355, SKB). Furthermore, the synthesis for the production of benztriazole was described by Katritzkv et al. in J. Org. Chem. 1997, 62, 4155-4158 and in J.Chem. Research (s) 1999, 230-231. Furthermore, carbamates are known as lipase inhibitors, such as Shamkant Patkar et al. in Paul Woolley, Steffen B. Petterson (ed), Lipase (1994) 207-227 or WO 03/051842.

Unexpectedly, it could now be shown that the benzotriazoles, which are defined according to the present invention, act on HSL, a hormone sensitive lipase.

This invention relates to benzotriazoles of general formula I,

in which they indicate:

R1 to R8 H,

whereby one of these residues R2 or R3 can stand in place of: Br, Cl, CH3, CN, NH2, NO2, CF3, OCH3, phenoxy, benzoyl,

CH(OH)-phenyl, S-cyclohexyl, CO-OCH3;

or

two substituents from this sequence are:

R1 = Cl and R3 = CF3 or

R2 = F and R3 - Cl;

n one integer 0, 1 or 2; and

one of the substituents R6 or R7 can stand for:

R6 CH3;

R7 CH3, C2H5; CH(CH3)2, C(CH3)3, CF3, Br, Cl, benzoyl or CO-OC2H5; or R6 and R7 are both CH3; or

the ring instead of R6 and R7 may contain one double bond or

R5 and R6 or R6 and R7 together with the C-atoms that carry them stand in place of one ring attached to benzene or if n = 0, it can stand in place of cyclohexanediyl, while in the case of ring closure R6/R7 this substituent can be substituted once with NH2

or NO 2 or once or twice with OCH 3 ; and

R 7 and R 8 together are cyclopentyl, diazirine or =CH 2 ;

the compounds with R1 to R5 and R8 = H, n = 1 and R6/R7 = benzene attached and R1, R3 -R8 - H, R2 = CH3 and n - 1 are excluded.

The invention relates to compounds of formula I, in the form of their racemates, racemic mixtures and pure enantiomers as well as their diastereomers and mixtures thereof.

Alkyl residues can be both linear and branched. Halogen stands in for fluorine, chlorine or bromine, especially fluorine or chlorine.

Priority is given to benzotriazoles of formula I,

in which they indicate:

R1 to R8 H,

whereby one of these R2 or R3 residues can stand in place of:

R2 Br, Cl, CN, NO2, CF3, OCH3, phenoxy, benzoyl, CH(OH)-phenyl,

S-cyclohexyl, CO-OCH3;

R3 CH3, CN, Br, Cl, NH2, NQ2, benzoyl.

Particular priority is given to benzotriazoles of formula I, in which they denote:

R1 to R8 are H;

whereby one of these R2 or R3 residues can stand in place of:

R2 Br, Cl, NO2, OCH3, phenoxy, CO-OCH3;

R3 NH2; or

two substituents from this sequence are:

R2 = F and R3 - Cl;

n is an integer 1 or 2; and

one of the substituents R5 or R6 can stand for:

R6 CH3;

R7 is CH3, CF3 or Br; or

R6 and R7 both residues are CH3; or

the ring instead of R6 and R7 may contain one double bond or

R6 and R7 together with the C-atoms they bear can stand for one ring attached to benzene, which in a given case can be substituted

once with NH2 or once to twice with OCH3; and

R7 and R8 together are cyclopentyl; or

n an integer 0; and

R6 and R7 together with the C-atoms they carry can stand for one benzene

attached ring or cyclohexanediyl; or

benzotriazoles of the general formula I, in which they denote

R1 to R8 are H;

whereby one of these R2 or R3 residues can stand in place of:

R2 Br, CN, CF3, OCH3, phenoxy, benzoyl, CH(OH)-phenyl, S-cyclohexyl; R3 CN, Br, Cl, NO2, benzoyl; or

two substituents from this sequence are:

R1 = Cl and R3 = CF3;

n is an integer 1; and

one of the substituents R6 or R7 can stand for:

R6 CH3;

R7 CH3, C2H5, CH(CH3)2, C(CH3)3, benzyl or CO-OC2H5; or

R6 and R7 both residues are CH3; or

the ring instead of R6 and R7 may contain one double bond or

R 5 and R 6 or R 6 and R 7 together with the C-atoms bearing them can stand for

one benzene ring attached;

compounds with R1 to R5 and R8 = H, n = 1 and R6/R7 = benzene attached are excluded.

Benzotriazoles with the following structures have a very special priority:

or benzotriazoles of the following structures:

Furthermore, benzotriazoles with the following structures have a very special priority:

and benzotriazoles of the following structures:

Pharmaceutically acceptable salts, based on their greater solubility in water compared to the starting or base compounds, are particularly suitable for use in medicine. These salts must have a pharmaceutically acceptable anion or cation. Considered as pharmaceutically acceptable salts of the compounds defined according to this disclosure are salts of inorganic acids, such as hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric and sulfuric acids, as well as organic acids, such as, for example, acetic, benzenesulfonic, benzoic, citric, ethanesulfonic, fumaric, gluconic, glycolic, isothionic, lactic, lactobionic, maleic, malic, methanesulfonic, succinic, p-toluenesulfonic and tartaric acids. Suitable pharmaceutically acceptable alkali salts are ammonia salts, alkali metal salts (such as sodium or potassium salts) and alkaline earth metal salts (such as magnesium and calcium salts), trometamol (2-amino-2-hydroxymethyl-1,3-propanediol), diethanolamine, lysine or ethylenediamine.

Salts with some anion, which is not pharmaceutically acceptable, such as trifluoroacetate, also fall within the scope of this invention as useful intermediates for the production or purification of pharmaceutically acceptable salts and/or for use in non-therapeutic, for example in vitro applications.

The term "physiologically functional derivative" as used herein means any physiologically acceptable derivative of a compound, which is defined according to the present invention, for example an ester, which when given to certain mammals, such as e.g. in humans, is able to be translated (directly or indirectly) into some compound of formula I or some active metabolite.

Physiologically functional derivatives also include prodrugs of compounds defined according to this invention, as described for example in H. Okada et al., Chem. Pharm. Bully. 1994, 42, 57-61. Such prodrugs can be metabolized in vivo to the compounds of the present invention. These prodrugs may or may not be active themselves.

The compounds defined according to this invention can exist in different polymorphic forms, for example in amorphous and in crystalline polymorphic forms. All polymorphic forms of the compounds defined according to this invention fall within the scope of this invention and represent another aspect of this invention. Hereinafter, all references to "compound/compounds of formula I" refer to compound/compounds of formula I as previously described, as well as their salts, solvates and physiologically functional derivatives as described herein.

The compound/compounds according to formula I can be dosed to patients in combination with other active substances.

The amount of a compound of formula I required to achieve the desired biological effect depends on a number of factors, for example on the specific compound chosen, on the intended use, on the method of administration and on the clinical condition of the patient. In general, the daily dose is in the range of 0.3 mg to 100 mg (typically 3 mg to 50 mg) per day per kilogram of body weight, for example 3-10 mg/kg/day. A single intravenous dose can be for example in the range of 0.3 mg to 1 mg/kg, most conveniently in the form of an infusion it can be dosed from 10 ng to 100 ng per kilogram per minute. Suitable infusion solutions for these purposes may contain for example from 0.1 ng to 10 mg, usually from 1 ng to 10 mg per milliliter. Individual doses can contain, for example, from 1 mg to 10 g of the active substance. Thus, ampoules for injection can contain, for example, from 1 mg to 100 mg, and single-dose formulations for oral use, such as tablets or capsules, can, for example, contain from 1.0 to 1000 mg, usually from 10 to 600 mg. In order to treat the above-mentioned conditions, the compounds according to formula I can be administered independently in the form of a compound, but primarily, however, they are found in a mixture with some acceptable carrier in the form of a single pharmaceutical preparation. The carrier must, of course, be acceptable in this sense, that it is compatible with the other ingredients of the preparation and that it is not harmful to the health of the patients. The carrier can be a solid substance or a liquid or both, and preferably a formulation is made with the compound in the form of a single dose, for example in the form of a tablet, which can contain from 0.05% to 95% by weight. % active substance. And other pharmaceutical active substances can also be found in the preparation, including other compounds according to formula I. Pharmaceutical preparations, which are defined according to this invention, can be produced according to one of the known pharmaceutical methods, which basically consists in the fact that the components are mixed with a pharmacologically acceptable carrier and/or auxiliary substances.

Pharmaceutical preparations according to the present invention are such that they are suitable to be dosed orally, rectally, topically, perorally (for example under the tongue) and parenterally (for example subcutaneously, intramuscularly, intradermally or intravenously), but the most suitable dosage method for each specific case depends on the type and severity of the condition to be treated, and on the type of compound used defined according to formula 1.1, the formulation in the form of dragees, as well as the formulation with prolonged action in the form of dragees, fall within the scope of this invention. Priority is given to formulations that are resistant to the action of acids and gastric juices. Suitable gastric-resistant dragee coatings include cellulose acetate phthalate, polyvinylacetate phthalate, hydroxypropyl methyl phthalate cellulose, and anionic polymers of methacrylic acid and methacrylic acid methyl ester.

Compounds contemplated to be used in pharmacy for oral dosage may be in individual units, such as in capsule form, capsules encapsulating drugs with a bad taste, lozenges or tablets, which always contain a certain amount of a compound according to formula I; in the form of powder or granules; in the form of a solution or suspension in an aqueous or non-aqueous liquid; or in the form of oil-in-water or water-in-oil emulsions. These preparations can, as already mentioned, be prepared using any suitable pharmaceutical method, which includes a step in which the active substance and the carrier (which may consist of one or more additional components) are brought into contact. In general, the preparations are produced by uniform and homogenizing mixing of the active substance with some liquid and/or finely dispersed solid carrier, after which, if necessary, the product is given a certain shape. Thus, for example, a tablet can be made, whereby the powder or granulate of the compound is pressed or shaped, in a given case with one or more additional components. Pressed tablets can be produced in a suitable machine by tableting the compound in a free flowing form, such as a powder or granulate, which is optionally mixed with a binder, a lubricant, an inert diluent and/or with one (more) surfactant/dispersant. Molded tablets may be produced in a suitable machine, whereby the compound in powder form is moistened with some inert liquid diluent.

Pharmaceutical preparations which are contemplated to be used for peroral dosing (under the tongue) include lozenges, which contain a compound of formula I together with some flavor enhancer, commonly sucrose and gum arabic or tragacanth, and pastilles, which contain this compound in some inert base, such as gelatin and glycerin or sucrose and gum arabic.

Pharmaceutical preparations contemplated for use for parenteral dosing include primarily sterile aqueous formulations of a compound of formula I, which are primarily isotonic with the blood of the intended recipient. These formulations are administered by injection primarily intravenously, but may also be administered subcutaneously, intramuscularly, or intradermally. These formulations can be produced primarily by mixing the compound with water and the resulting solution is sterilized and made isotonic with blood. Formulations defined according to the present invention, which can be given in the form of injections, generally contain from 0.1 to 5 wt. % of active compound.

Pharmaceutical preparations, which are considered to be given to patients through the rectum, are present primarily in the form of single doses, in the form of suppositories. These can be produced by mixing the compound of formula I with one or more conventional solid carriers, for example cocoa butter, and forming the resulting mixture into suppositories.

Pharmaceutical preparations that come into consideration for topical application to the skin are primarily in the form of salves, creams, lotions, pastes, sprays, aerosols or oils. Vaseline, lanolin, polyethylene glycols, alcohols or combinations of two or more of these substances can be used as carriers. The active substance is generally found in a concentration of 0.1 to 15 wt.% of the preparation, for example from 0.5 to 2 wt. %.

It is also possible to use the preparation transdermally. Pharmaceutical preparations suitable for transdermal applications may be in the form of individual patches, which are suitable for close contact with the patient's epidermis over a long period of time. Such patches contain an active substance in a suitable form, in a given case in a buffered aqueous solution, dissolved and/or dispersed in an adhesive agent or dispersed in a polymer. A suitable concentration of the active substance is about 1% to 35%, preferably about 3% to 15%. One special possibility is that the active substance can, as described for example in Pharmaceutical Research, 2(6): 318 (1986), be released via electrophoresis or iontophoresis.

The following are suitable as other active substances for combined preparations:

all antidiabetics, which are listed in the Red List 2001, chapter 12. They can be combined with the compounds of formula I, defined according to the present invention, in particular in order to synergistically improve the effect. Dosing of a combination of active substances can take place by separately administering the active substances to the patient or by administering them in the form of combined preparations, whereby several active substances are present in one pharmaceutical preparation. The largest number of active substances, which are listed below, were published in the USP Dictionarv of USAN and International Drug Names, US Pharmacopeia, Rockville 2001.

Antidiabetics include insulin and insulin derivatives, such as Lantus<®> (see www.lantus.com) or HMR 1964, rapid-acting insulins (see US, 6,221,633), GLP-1 derivatives, such as those disclosed by Novo Nordisk A/S in patent WO 98/08871, as well as oral hypoglycemic active substances. Hypoglycemic active substances, which act orally, include primarily sulfonylureas, biguanidines, meglitinides, oxadiazolidinediones, thiazolidinediones, glucosidase inhibitors, glucagon antagonists, GLP-1 agonists, potassium channel openers, such as those published by Novo Nordisk A/S in WO 97/26265 and WO 99/03861, insulin sensitizers, inhibitors liver enzymes, which take part in the stimulation of gluconeogenesis and/or glycogenolysis, modulators of glucose binding, compounds that change fat metabolism such as antihyperlipidemic active substances and antilipidemic active substances, compounds that affect the reduction of food intake, PPAR- and PXR - agonists and active substances that act on the ATP-dependent potassium channel of beta cells.

In one embodiment of the present invention, compounds of formula I are applied in combination with an HMGCoA-reductase inhibitor such as simvastatin, fluvastatin, pravastatin, lovastatin, atorvastatin, cerivastatin, rosuvastatin.

In one embodiment of the present invention, compounds of formula I are administered in combination with a cholesterol resorption inhibitor, such as ezetimibe, tiquesid, pamaquesid.

In one embodiment of the present invention, compounds of formula I are dosed in combination with a PPAR gamma agonist, such as, for example, rosiglitazone, pioglitazone, JTT-501, GI 262570.

In one embodiment of the present invention, compounds of formula I are used in combination with a PPAR alpha agonist, such as, for example, GW 9578, GW 7647.

In one embodiment of the present invention, compounds of formula I are administered in combination with some mixed PPAR alpha/gamma agonists, such as for example GW 1536, AVE 8042, AVE 8134, AVE 0847 or as described in WO 00/64888, WO 00/64876, WO 03/020269.

In one embodiment of the present invention, the compounds of formula I are dosed in combination with a fibrate, such as for example with fenofibrate, with clofibrate, with bezafibrate.

In one embodiment of this invention, compounds of formula I are applied in combination with some MTP inhibitor, such as, for example, implitapide, BMS-201038, R-103757.

In one embodiment of the present invention, compounds of formula I are administered in combination with a bile acid resorption inhibitor (see for example US 6,245,744 or US 6,221,897), such as for example HMR 1741.

In one embodiment of the present invention, compounds of formula I are dosed in combination with a CETP inhibitor, such as JTT-705.

In one embodiment of the present invention, the compounds of formula I are administered in combination with a polymeric bile acid adsorbent, such as, for example, cholestyramine, colesevelam.

In one embodiment of the present invention, compounds of formula I are administered in combination with an LDL-receptor inducer (see US 6,342,512), such as HMR1171, HMR1586.

In one embodiment of the present invention, the compounds of formula I are administered in combination with an AC AT-inhibitor, such as, for example, avasimib.

In one embodiment of the present invention, compounds of formula I are used in combination with an antioxidant, such as OPC-14117.

In one embodiment of the present invention, compounds of formula I are dosed in combination with a lipoprotein lipase inhibitor, such as NO-1886.

In one embodiment of the present invention, the compounds of formula I are administered in combination with an ATP-citrate-lyase inhibitor, such as SB-204990.

In one embodiment of the present invention, compounds of formula I are administered in combination with a squalene synthetase inhibitor, such as BMS-188494. In one embodiment of the present invention, compounds of formula I are dosed in combination with a lipoprotein antagonist, such as CI-1027 or nicotinic acid.

In one embodiment of the present invention, the compounds of formula I are administered in combination with a lipase inhibitor, such as orlistat.

In one embodiment of the present invention, the compounds of formula I are administered in combination with insulin.

In one embodiment, the compounds of formula I are prescribed in combination with a sulphonylurea, such as for example tolbutamide, glibenclamide, glipizide or glimepiride.

In one embodiment, compounds of formula I are administered in combination with a biguanide, such as, for example, metformin.

In another embodiment, the compounds of formula I are administered in combination with a meglitinide, such as repaglinide.

In one embodiment, the compounds of formula I are administered in combination with a thiazolidinedione, such as for example troglitazone, ciglitazone, pioglitazone, rosiglitazone or the compounds disclosed in patent WO 97/41097 by Dr. Reddv's Research Foundation, particularly the compound 5-[[4-[(3,4-dihydro-3-methyl-4-oxo-2-quinazolinyl-methoxy]phenyl]methyl]-2,4-thiazolidinedione.

In one embodiment, compounds of formula I are administered in combination with an α-glucosidase inhibitor, such as miglitol or acarbose.

In one embodiment, compounds of formula I are administered in combination with an active substance that acts on the ATP-dependent potassium channel of beta cells, such as for example tolbutamide, glibenclamide, glipizide, glimepiride or repaglinide.

In one embodiment, compounds of formula I are dosed in combination with more than one of the above-mentioned compounds, for example in combination with some sulfonylurea and metformin, with some sulfonylurea and acarbose, with repaglinide and metformin, with insulin and some sulfonylurea, with insulin and metformin, with insulin and troglitazone, with insulin and lovastatin, etc.

In one of the other embodiments, compounds of formula I are applied in combination with CART modulators (see "Cocaine-amphetamine-regulated transcript influences energy metabolism, anxiety and gastric emptying in mice" Asakawa, A. et al., M: Hormone and Metabolic Research (2001), 33 (9), 554 - 558), NPY-agonists, for example naphthalene-1-sulfonic acid {4-[(4- amino-quinazolin-2-ylamino)-methyl]-cyclohexylmethyl}-amide; hydrochloride (CGP 71683A)), MC4 - agonists (for example 1-amino-1,2,3,4-tetrahydro-naphthalene-2-carboxylic acid [2-(3a-benzyl-2-methyl-3-oxo-1,3,3a,4,6,7-hexahydro-pyrazolo[4,3-c]pyridin-5-yl)-1-(4-chloro-phenyl)-2-oxo- ethyl]-amide; (WO 01/91752), orexin-agonists (for example 1-(2-methyl-benzoxazol-6-yl)-3-[1,5]naphthyridin-4-yl-urea; hydrochloride (SB-334867-A)), H3-agonists (3-cyclohexyl-1-(4,4-dimethyl -1,4,6,7-tetrahydro-imidazo[4,5]pyridin~5-yl)- propan-l-one salt of oxalic acid (WO 00 / 63208)); TNF - agonists, CRF - agonists (for example [2-methyl-9-(2,4,6-trimethyl-phenyl)-9H-1,3,9-triaza-fluoren-4-yl]-dipropyl-amine (WO 00 / 66585)), CRF BP - antagonists (for example urocortin), urocortin-agonists, JB-agonists (for example l-(4-chloro-3-methanesulfonylmethyl- phenyl)-2-[2-(2,3-dimethyl-1H-indol-6-yloxy)-ethylaminoj-ethanol; hydrochloride (WO 01/83451)), MSH (melanocyte-stimulating hormone)-agonists, CCK-A agonists (for example {2-[4-(4-chloro-2,5- dimethoxy-phenyl)-5-(2-cyclohexyl-ethyl)-triazol-2-ylcarbamoyl]-5,7-dimethyl-indol-1-yl}-acetic acid with trifluoroacetic acid (WO 99 / 15525)); serotonin reuptake inhibitors (e.g. dexfenfluramine), a mixture of serotonin- and noradrenostimulatory compounds (e.g. WO 00/71549), 5HT-agonists e.g. l-(3-ethyl-benzofuran-7-yl)-piperazine salt oxalic acid (WO 01/ 09111), bombesin agonists, galanin antagonists, growth hormone (e.g. human growth hormone), hormone-releasing compounds growth (6-benzyloxy-1-(2-diisopropylamino-ethylcarbamoyl)-3,4-dihydro-1H-isoquinoline-2-carboxylic acid-t-butyl-ester (WO 01/ 85595), TRH agonists (see for example EP 0 462 884), 2- or 3-modulators that break down protein, Ieptin agonists (see for example Lee, Daniel W.; Leinung, Matthew C.; Rozhavskaya-Arena, Marina; Grasso, Patricia: Leptin agonists a potential approach to the treatment of obesity. Drugs of the Future

(2001), 26 (9), 873 - 881), DA agonists (bromocriptine, doprexin), lipase/amylase inhibitors (eg WO00/40569), PPAR-modulators (eg WO 00/78312), RXR-modulators or TR-/3-agonists.

In one embodiment of the present invention, leptin is the second active substance; see for example "Perspectives in the therapeutic use of leptin", Salvador, Javier; Gomes-Ambrosi, Javier; Fruhbeck, Gemma: Expert Opinion on Pharmacotherapy (2001), 2 (10), 1615 - 1622.

In one embodiment, the second active substance is dexamfetamine or amphetamine. In one embodiment, the second active substance is fenfluramine or dexfenfluramine. In another embodiment, sibutramine is the second active substance.

In one embodiment, orlistat is the second active substance.

In one embodiment, the second active substance is mazindol or phentermine.

In one embodiment, compounds of formula I are administered to the patient in combination with bulking substances, primarily with insoluble bulking substances (see for example Carob/ Caromax<®>(Zunft H. J., et al. Carob pulp preparation for treatment of hvpercholesterolemia, ADVANCES IN THERAPY (2001 Sep-Oct), 18 (5), 230 - 6). Caromax, which contains carob, is a product of Fa. Nutrinova, Nutrition Specialties & Food Ingredients GmbH, Industriepark Hoechst, 65926 Frankfurt/Main). The combination with the substance Caromax<®> can be made in one preparation, but the compounds of formula I and the substance Caromax<®> can also be dosed separately. Caromax<®> can also be given in the form of foodstuffs, such as in pastries or in Muslim.

It is understood that any suitable combination of compounds defined according to this invention, with one or more of the above-mentioned compounds and optionally with one or more other pharmacologically active substances, also falls within the scope of patent protection of this invention. The production of benzotriazoles of the general formula I, which are defined according to this invention, takes place according to known methods, for example by applying acylation of substituted or unsubstituted benzotriazole 2 with carbamoyl chloride 3 (method A), or in two stages by conversion of benzotriazole with phosgene and further reaction of the resulting benzotriazole-carboxylic acid-chloride with amines (method

B).

Since acids are usually released during these reactions, it is recommended to add bases such as pyridine, triethylamine, sodium hydroxide or alkali carbonates in order to speed up the reaction. Reactions can take place in a wide temperature range. As a rule, it has been shown to be advantageous to operate at 0 °C up to the boiling point of the solvent used. As a solvent, examples of methylene chloride, THF, DMF, toluene, acetic acid ester, n-heptane, dioxane, diethyl ether can be used.

The compounds of general Formula 1, which are defined according to this invention, have an unexpected inhibitory effect on hormone-sensitive lipase, HSL, an allosteric enzyme in fat cells, which is inhibited by insulin and which is responsible for the breakdown of fat in fat cells, and therefore for the transfer of fat components into the blood vessel system. Inhibition of this enzyme by the action of the compounds defined in this article therefore corresponds to an action similar to the action of insulin, which ultimately leads to a reduction of free fatty acids in the blood and to a reduction of blood sugar. They can therefore be used for metabolic irregularities such as diabetes mellitus, which is not dependent on insulin, for diabetic syndrome, for syndrome X and for direct damage to the pancreas.

Some inhibition of HSL in P-cells should lead to a direct recovery of insulin release (M. Winzell et al., Diabetes, vol 52, August 2003, 2057-2065). The compounds of formula I defined according to the present invention can therefore also serve to release insulin.

The activity of the compounds of formula I, which are defined according to the present invention, was tested on the following enzyme test system:

Substrate preparation:

Preparation of NAG (NBD - monoacylglyceride) substrate:

6 mg of phosphatidylcholine and 6 mg of phosphatidylinositol are dissolved in 1 mL of chloroform each. 10 mg of NAG was dissolved in 1 mL of chloroform. Two parts of the phosphatidylinositol solution (eg 83.5 (µL) and one part of the phosphatidylcholine solution (eg 41.5 µL) and 100 µL of the NAG solution are pipetted together into a plastic scintigraphy dish (final assay concentration: 0.0375 mg phospholipid/mL; 0.05 mg/NAG/mL). Chloroform (225 µL total volume) is removed by blowing with one stream. N2. The dried substrate can be stored for up to 3 days at 4 °C. In order to produce phospholipid vesicles/micelles with inserted NAG (on the day of testing), the dried substrate is placed in 20 mL of research buffer (25 mM Tris/HCl, pH 7.4; 150 mM NaCl) and two sonications are performed using a single ultrasonic wand (Branson Sonifier Type II, standard microtip): 1. 2.2x1 min, meanwhile always 1 min on ice; 2. Processing position 4, 2 x 1 min, in the meantime always 1 min on ice. During this procedure, the color of the substrate solution changes from yellow (extinction maximum 481 nm) to red (extinction maximum 550 nm) due to the insertion of NAG between the phospholipid molecules of vesicles/micelles. Before using as a substrate (for the next 2 h), the solution is incubated on ice for another 15 min.

Indirect NAG research:

The research is carried out in Eppendorf vessels with a volume of 1.5 mL or in 96-well plates at a temperature of 30 °C. In order to determine HSL inhibitors, 10 µL of the test substance is previously placed in the research buffer (25 mM Tris/HCl, pH 7.4; 150 mM NaCl) in the presence of 16.6% DMSO. 180 µL of substrate solution (20 µg/mL phosphatidylcholine, 10 µg/mL phosphatidylinositol, 50 µg/mL NAG in research buffer) is added to it. After one pre-incubation for 15 min at 30 °C, 20 µL of enzyme solution in research buffer (1- to 4-fold diluted) is pipetted into it and the extinction is immediately measured at 480 nm in in a photometer with cuvettes (0.5 mL cuvettes) or in a device with microtiter plates. After incubation for 60 min at 30 °C, the extinction is measured at 480 nm. Under standard conditions, 20 µg of partially purified

HSL to extinction change of 0.4 = 4000 arbitrage units.

Direct NAG research:

As an alternative to measuring the change in extinction of the substrate solution, the products of the HSL reaction are examined by phase separation/thin layer chromatography. To this end, an incubation batch (200 µL total volume, see indirect NAG research) was mixed in a 2 mL Eppendorf dish with 1.3 mL methanol/chloroform/heptane (10:9:7) and finally with 0.4 mL 0.1 M NaOH. After intense mixing (10 sec), the phases are separated by centrifugation (800 x g, 20 min, room temperature). Equivalent volumes (for example 0.4 mL) are taken from the aqueous upper phase and the extinction is determined photometrically at 481 nm. For thin layer chromatography, the mobile phase was dried (SpeedVac) and then mixed in 50 µL of tetrahydrofuran. Samples with a volume of 5 µL were applied to silica gel Si-60 plates (Merck). Chromatography is performed using a mixture of 78 mL of diethyl ether/22 mL of petroleum ether/1 mL of glacial acetic acid as an eluent. The amount of released fluorescent NBD-fatty acid is determined by phosphorimaging (Molecular Dynamics, Storm 840 and ImageQuant Software) at an excitation wavelength of 460 nm and an emission wavelength of 540 - 560 nm.

Enzyme preparation:

Preparation of partially purified HSL:

Isolated rat fat cells were obtained from epididymal adipose tissue of untreated male rats (Wistar, 220-250 g) using collagen treatment according to a published procedure (eg S. Nilsson et al., Anal. Biochem. 158, 1986, 399-407; G. Fredikson et al., J. Biol. Chem. 256, 1981, 6311 - 6320; H. Tomquist, J. Biol. 251, 813-819). Fat cells from 10 rats were washed three times by flotation each time with 50 mL homogenization buffer solution (25 mL tris/HCl, pH 7.4, 0.25 M sucrose, 1 mM EDTA, 1 mM DTT, 10 µg/mL Leupeptin, 10 µg/mL antipain, 20 µg/mL pepstatin) and finally placed in 10 mL buffer solution for homogenization. Fat cells are homogenized in a Teflon-in-glass homogenizer (Braun-Melsungen) with 10 elevations at 1500 revolutions per minute and at a temperature of 15 °C. The homogenate is centrifuged (Sorvall SM24 tubes, 5000 revolutions per minute, 10 min, 4 °C). The bottom layer between the fat layer, which lies above, and the pellet is separated and the centrifugation is repeated. The lower layer resulting from this centrifugation is separated and centrifuged again (Sorvall SM24 tube, 20000 revolutions per minute, 45 min, 4 °C). The lower layer is separated and treated with 1 g of heparin-sepharose (Pharmacia Biotech, C1-6B, washed 5 times with 25 mM Tris/HCl, pH 7.4, 150 mM NaCl). After incubation for 60 min at a temperature of 4 °C (stirring at 15 min intervals), the sediment is centrifuged (Sorvall SM24 tubes, 3000 revolutions per minute, 10 min, 4 °C). The lower layer is adjusted to pH 5.2 by adding glacial acetic acid and incubated for 30 minutes at a temperature of 4 °C. The precipitates were collected by centrifugation (Sorvall SS34, 12000 rpm, 10 min, 4 °C) and their suspension was made in 2.5 mL of 20 mM Tris/HCl, pH 7.0, 1 mM EDTA, 65 mM NaCl, 13% sucrose, 1 mM DTT, 10 µg/mL leupeptin/pepstatin/antipain. This suspension is dialyzed overnight at 4 °C with 25 mM Tris/HCl, pH 7.4, 50% glycerin, 1 mM DTT, 10 µg/mL leupeptin, pepstatin, antipain and then applied to a single hydroxyapatite column packing (0.1 g per 1 mL suspension, equilibrated with 10 mM potassium phosphate, pH 7.0, 30% glycerin, 1 mM DTT). The column pack is washed with four volumes of equilibration buffer solution at a flow rate of 20 to 30 mL/h. HSL is eluted with one volume of equilibration buffer containing 0.5 M potassium phosphate, then dialyzed (see above) and concentrated 5- to 10-fold using ultrafiltration (Amicon Diaflo PM 10 Filter) at 4 °C. Partially purified HSL can be stored for 4 to 6 weeks at -70 °C.

Examination:

In order to produce the substrate, 25 - 50 uCi of [<3>H Jtrioleoylglycerol (in toluene), 6.8 uMol of unlabeled trioleoylglycerol and 0.6 mg of phospholipid (phosphatidylcholine / phosphatidylinositol 3 : 1 mass / volume) are mixed, dried in an N2 atmosphere and then dispersed in 2 mL of 0.1 M KPi (pH 7.0) by sonication (Branson 250, micro tip, setting 1-2, 2 x 1 min at 1 minute intervals). After adding 1 mL of KPi and re-treatment with ultrasound (4 x 30 sec on ice and intervals of 30 sec), 1 mL of 20% BSA (in KPi) is added (final concentration of trioleoylglycerin is 1.7 mM). In order to perform the reaction, 100 uL of substrate solution is taken with a pipette and added to 100 uL of HSL solution (HSL prepared as above, diluted in 20 mM KPi, pH 7.0, 1 mM EDTA, 1 mM DTT, 0.02% BSA, 20 µg/mL pepstatin, 10 µg/mL leupeptin) and incubated for 30 min at a temperature of 37 °C. After adding 3.25 mL of a mixture of methanol/chloroform/heptane (10 : 9 : 7) and 1.05 mL of 0.1 M K2C03, 0.1 M boric acid (pH 10.5) was mixed well and finally centrifuged (800 x g, 20 min). After separating the phases, take one equivalent of the upper phase (1 mL) and determine the radioactivity using the liquid scintillation method.

Calculation:

Substances are usually tested in four independent runs. The inhibition of HSL enzyme activity caused by the application of the test substance is determined by comparison with a control reaction that is not inhibited. The calculation of the IC50-value takes place according to one curve, which shows the inhibitions with at least 10 concentrations of the test substance. The software package GRAPHIT, Elsevier - BIOSOFT, was used for data analysis.

In this test, the compounds from examples 1 to 55 showed inhibitions in the IC50 range of 0.04 - 5 µM.

The following examples serve to explain this invention in more detail, but not to limit the invention thereby.

Examples:

The following examples were produced according to the methods described in the following text:

Method A:

To a solution containing 2 mmol of 1H-benzotriazole in pyridine (5 mL) and dichloromethane (10 mL) was added a solution of the corresponding carbamoyl chloride (1 mmol) in dichloromethane (10 mL). The reaction mixture was stirred for 16 h at room temperature, then mixed with EtOAc (15 mL), filtered over silica gel and the filtrate concentrated. The product is purified using preparative HPLC and dried using the lyophilization procedure.

Method B:

a) Production of one solution of benzotriazole-l-carboxylic acid-chloride One solution of phosgene (20% in toluene; 90 mL; 182 mmol) is added dropwise with cooling

with ice a solution of benzotriazole (6 g, 50.4 mmol) in THF (100 mL). The ice bath was removed and the solution was stirred for another 2 h at room temperature. It was dissolved away by distillation and the residue was mixed in THF to a total volume of 25 mL.

b) Conversion of benzotriazolecarboxylic acid-chlorides into corresponding benzotriazole-l-carboxylic acid-amides and anilides

Each time 10 mg of amine or aniline (2 mmol) was placed in THF (1 mL) and pyridine (0.2 mL) was added. The batch was incubated with a solution of benzotriazole-1-carboxylic acid chloride (1 mL, 2 mmol) and stirred continuously for 16 h at room temperature. At the end, the batches are diluted with ethyl acetate (5 mL), filtered through silica gel, and the filtrates are concentrated to dryness in a vacuum. The crude products are purified using flash chromatography.

Example 1:

3-(4-methyl-piperidine-1-carbonyl)-3H-benzotriazole-5-carboxylic acid-methyl ester

M + H+: 303.14

Example 2:

(8-aza-spiro[4,5]dec-8-yl)-benzotriazol-1-yl-methanone

M + H+: 285.16

Example 3:

Benzotriazol-1-yl-(6,7-dimethoxy-3,4-dihydro-1H-isoquinolin-2-yl)-methanone

M + H+: 339.13

Example 4:

(5-phenoxy-benzotriazol-1-yl)-(4-trifluoromethyl-piperidin-1-yl)-methanone

M + H+: 391.13

Example 5:

(6-chloro-5-fluoro-benzotriazol-1-yl)-(4-methyl-piperidin-1-yl)-methanone

M + H<+>: 297.74

Example 6:

Benzotriazol-1-yl-(4-bromo-piperidin-1-yl)-methanone

M + H+: 310.3

Example 7:

Benzotriazol-1-yl-(4-trifluoromethyl-piperidin-1-yl)-methanone

M + H+: 299.18

Example 8:

benzotriazol-1-yl-(1,3-dihydro-isoindol-2-yl)-methanone

M + H+: 265.0

Example 9:

1-(3,6-dihydro-2H-pyridine-1-carbonyl)-1H-benzotriazole-5-carboxylic acid methyl ester

M + H+: 287.04

Example 10:

(3,6-dihydro-2H-pyridin-1-yl)-(5-nitro-benzotriazol-1-yl)-methanone

M + Na: 296.21

Example 11:

(3,4-dihydro-1H-isoquinolin-2-yl)-(5-nitro-benzotriazol-1-yl)-methanone

M + H+: 324.10

Example 12:

(5-bromo-benzotriazol-1-yl)-(3,6-dihydro-2H-pyridin-1-yl)-methanone

M + H4": 306.98

Example 13:

(5-bromo-benzotriazol-1-yl)-(3-trifluoromethyl-piperidin-1-yl)-methanone

M + H+: 377.30

Example 14:

(5-bromo-benzotriazol-1-yl)-(3,4-dihydro-1H-isoquinolin-2-yl)-methanone

M + H<+>: 357.04

Example 15:

Benzotriazol-1-yl-(octahydro-isoindol-2-yl)-methanone

M + FT": 271.15

Example 16:

(7-amino-3,4-dihydro-1H-isoquinolin-2-yl)-benzotriazol-1-yl-methanone

M + H+: 294.0

Example 17:

(3,4-dihydro-1H-isoquinolin-2-yl)-(5-methoxy-benzotriazol-1-yl)-methanone

M + H<+>: 309.04

Example 18:

(5-Methoxy-benzotriazol-1-yl)-(3-methyl-piperidin-1-yl)-methanone

M + H<+>: 275.5

Example 19:

(6-amino-benzotriazol-1-yl)-(4-methyl-piperidin-1-yl)-methanone

M + H+: 260.1

Example 20:

(5-chloro-benzotriazol-1-yl)-(4-methyl-piperidin-1-yl)-methanone

M + H<+>: 279.6

Example 21:

Benzotriazol-1-yl-(1,2,6-triaza-spiro[2,5]oct-1-en-6-yl)-methanone

M + Na+: 279.19

Example 22:

Benzotriazol-1-yl-(4-ethyl-piperidin-1-yl)-methanone

M + H<+>: 259.04

Example 23:

(4-methyl-piperidin-1-yl)-(6-nitro-benzotriazol-1-yl)-methanone

M + H<+>: 290.4

Example 24:

1-(benzotriazole-1-carbonyl)-piperidine-4-carboxylic acid - ethyl ester

M + H+: 303.13

Example 25:

Benzotriazol-1-yl-(4-methyl-piperidin-1-yl)-methanone

M + H<+>: 245.0

Example 26:

3-(4-methyl-piperidine-1-carbonyl)-3H-benzotriazole-5-carbonitrile

M + H+: 270.12

Example 27:

Benzotriazol-1-yl-(3,4-dihydro-1H-isoquinolin-1-yl)-methanone

M + H+: 279.11

Example 28:

Benzotriazol-1-yl-(3,4-dihydro-2H-quinolin-1-yl)-methanone

M + H<+>: 279.2

Example 29:

(6-methyl-benzotriazol-1-yl)-pyrrolidine-1-yl-methanone

M + H+: 231.11

Example 30:

Benzotriazol-1-yl-(3-methyl-piperidin-1-yl)-methanone

M + H+: 245.13

Example 31:

Benzotriazol-1-yl-(3,4-dimethylpiperidin-1-yl)-methanone

M + H+: 259.14

Example 32:

[1-(4-methyl-piperidine-1-carbonyl)-1H-benzotriazol-5-yl]-phenyl-methanone

M + H+: 349.15

Example 33:

(4-methyl-piperidin-1-yl)-(5-trifluoromethyl-benzotriazol-1-yl)-methanone

M + H+: 313.5

Example 34:

(6-bromo-benzotriazol-1-yl)-(4-methyl-piperidin-1-yl)-methanone

M + H<+>: 324.0

Example 35:

Benzotriazol-1-yl-(4-t-butyl-piperidin-1-yl)-methanone

M + H+: 287.17

Example 36:

[5-(hydroxy-phenyl-methyl)-benzotriazol-1-yl]-(4-methyl-piperidin-1-yl)-methanone

M + H+: 350.17

Example 37:

(4-methyl-piperidin-1-yl)-(5-phenoxy-benzotriazol-1-yl)-methanone

M + H<+>: 337.3

Example 38:

(5-cyclohexylsulfanyl-benzotriazol-1-yl)-(4-methyl-piperidin-1-yl)-methanone

M + H+: 359.17

Example 39:

Benzotriazol-1-yl-(4-isopropyl-piperidin-1-yl)-methanone

M + H+: 273.3

Example 40:

(6-chloro-benzotriazol-1-yl)-(methyl-piperidin-1-yl)-methanone

M + H<+>: 279.5

Example 41:

Benzotriazol-1-yl-(6-methoxy-3,4-dihydro-1H-isoquinolin-2-yl)-methanone

M + H<+>: 309.3

Example 42:

Benzotriazol-1-yl-(4-benzyl-piperidin-1-yl)-methanone

M + H+: 321.1

Example 43:

(5-bromo-benzotriazol-1-yl)-(3-methyl-piperidin-1-yl)-methanone

M + H+: 325.31

Example 44:

1-(3,4-dihydro-1H-isoquinoline-2-carbonyl)-1H-benzotriazole-5-carbonitrile

M + H1": 270.12

Example 45:

(4-chloro-6-trifluoromethyl-benzotriazol-1-yl)-(3,4-dihydro-1H-isoquinolin-2-yl)-methanone

M + H+: 381.06

Example 46:

1-(3,4-dihydro-2H-quinoline-1-carbonyl)-1H-benzotriazole-5-carbo-nitrile

M + H+: 304.11

Example 47:

(4-chloro-6-trifluoromethyl-benzotriazol-1-yl)-(3,6-dihydro-2H-pyridin-1-yl)-methanone

M + H+: 331.04

Example 48:

Benzotriazol-1-yl-(7-methoxy-3,4-dihydro-1H-isoquinolin-2-yl)-methanone

M + H+: 309.1

Example 49:

(5-Methoxy-benzotriazol-1-yl)-(4-methyl-piperidin-1-yl)-methanone

M + H<+>: 275.3

Example 50:

(3,4-dihydro-1H-isoquinolin-2-yl)-(6-nitro-benzotriazol-1-yl)-methanone

M + H<+>: 324.3

Example 51:

(6- Benzoyl- benzotriazol-1-yl)-(4- methyl- piperidin-1-yl)-methanone

M + H+: 349.15

Example 52:

Benzotriazol-1-yl-(7-nitro-3,4-dihydro-1H-isoquinolin-2-yl)-methanone

M + H+: 324.1

Example 53:

Azepan-1-yl-benzotriazol-1-yl-methanone

M + H<+>: 245.3

Example 54:

Benzotriazol-1-yl-(4-chloro-piperidin-1-yl)-methanone

M + H+: 265.7

Example 55:

Benzotriazol-1-yl-(3,6-dihydro-2H-pyridin-1-yl)-methanone

M + H<+>: 229.2

Claims (12)

1. Benzotriazoles of general formula I, characterized by the fact that they denote: R1 to R8 H, whereby one of these residues R2 or R3 can stand in place of: Br, Cl, CH3, CN, NH2, NO2, CF3, OCH3, phenoxy. benzoyl, CH(OH)-phenyl, S-cyclohexyl, CO-OCH3; or two substituents from this sequence are: R1 = C1 and R3 = CF3 or R2 = F and R3 = Cl; n one integer 0, 1 or 2; and one of the substituents R 6 or R 7 can stand in place of: R 6 CH 3 ; R7 CH3, C2H5; CH(CH3)2. C(CH3)3. CF3. No. Cl. benzoyl or CO-OC2H3; or R6 and R7 are both CH3; or the ring instead of R 6 and R 7 may contain one double bond or R 5 and R 6 or R 6 and R 7 together with the C-atoms on which they are located stands for one ring attached to benzene or if n = 0, can stand in place of cyclohexanediyl, while in the case of ring closure R6/R7 this substituent can be substituted once with NH 2 or NO 2 or once or twice with OCH 3 ; and R 7 and R 8 together are cyclopentyl, diazirine or =CH 2 ; compounds with R1 to R5 and R8 = H, n = 1 and R6/R7 = joined to benzene and R1, R3 - R8 - H, R2 = CH3 and n = 1 are excluded. 2. Benzotriazoles of the general formula I, which are defined according to patent claim 1, characterized by the fact that they denote: R1 to R8 H; whereby one of these R2 or R3 residues can stand in place of: R2 Br, Cl, CN, NO2, CF3, OCH3, phenoxy, benzoyl, CH(OH)-phenyl, S-cyclohexyl, CO-OCH3; R3 CH3, CN, Br, Cl, NH2, NO2, benzoyl. 3. Benzotriazoles of the general formula I, which are defined according to patent claim 1 or 2, characterized by the fact that they denote: R1 to R8 H; wherein one of these residues R2 or R3 can stand in place of: R2 Br, Cl, NO2, OCH3, phenoxy, CO-OCH3; R3 NH2; or two substituents from this sequence are: R2 = F and R3 = Cl; n one integer 1 or 2; and one of the substituents R 5 or R 6 can stand in place of: R 6 CH 3 ; R7 is CH3, CF3 or Br; or the ring instead of R6 and R7 may contain one double bond or R6 and R7 together with the C-atoms on which they are located may stand in place of one benzene-fused ring, which in a given case can be substituted once with NH 2 or once to twice with OCH 3 ; and R 7 and R 8 together are cyclopentyl; or n one integer 0; and R 6 and R 7 together with the C-atoms on which they are located can stand for one ring fused to benzene or cyclohexanediyl. 4. Benzotriazoles of the general formula I, which are defined according to patent claims 1 to 2, characterized in that they denote R 1 to R 8 H; wherein one of these residues R2 or R3 can stand in place of: R2 Br, CN, CF3, OCH3, phenoxy, benzoyl, CH(OH)-phenyl, S-cyclohexyl; R3 CN, Br, Cl, NO2, benzoyl; or two substituents from this sequence are: R1 = Cl and R3 = CF3 ; n one integer 1; and one of the substituents R 6 or R 7 can stand in place of: R 6 CH 3 ; R7 CH3, C2H5, CH(CH3)2, C(CH3)3, benzyl or CO-OC2H5; or R6 and R7 are both CH3; or the ring instead of R6 and R7 may contain one double bond or R5 and R6 or R6 and R7, together with the C-atoms on which they occur, may stand instead of one benzene-fused ring; compounds with R1 to R5 and R8 = H, n = 1 and R6/R7 = benzene together are excluded. 5. Benzotriazoles according to claim 1, with the following structures: 6. Benzotriazoles according to claim 1, with the following structures: 7. Benzotriazoles of the following structures according to claim 6: 8. Benzotriazoles of the following structures according to claim 7: 9. Benzotriazoles of general formula I, in which they denote: R1 to R8 H, whereby one of these residues R2 or R3 can stand in place of: Br, Cl, CH3, CN, NH2, NO2, CF3, OCH3, phenoxy, benzoyl, CH(OH)-phenyl, S-cyclohexyl, CO-OCH3; or two substituents from this sequence are: R1 = Cl and R3 = CF3 or R2 = F and R3 = Cl; n one integer 0, 1 or 2; and one of the substituents R 6 or R 7 can stand in place of: R 6 CH 3 ; R7 CH3, C2H5; CH(CH3)2, C(CH3)3, CF3, Br, Cl, benzoyl or CO-OC2H5; or R6 and R7 are both CH3; or the ring instead of R6 and R7 may contain one double bond or R5 and R6 or R6 and R7, together with the C-atoms on which they are located, stand for one ring connected to benzene or if n = 0, it can stand in place of cyclohexanediyl, while in the case of ring closure R6/R7 this the substituent may in a given case be substituted once with NH 2 or NO 2 or once or twice with OCH 3 ; and R 7 and R 8 together are cyclopentyl, diazirine or =CH 2 ; indicated by the fact that they are used in some medication. 10. Benzotriazoles of formula I, which are defined according to patent claim 9, for use in one medication with an inhibitory effect on hormone-sensitive lipase, HSL. 11. Benzotriazoles of formula I, which are defined according to claim 9, for use in a single medicament for the treatment of non-insulin dependent diabetes mellitus, diabetic syndrome or syndrome X. 12. Medication for the treatment of non-insulin-dependent diabetes mellitus or diabetic syndrome containing at least one benzotriazole of formula I, which is defined according to patent claim 9.