KR20010080428A - Glucose and Lipid Lowering Compounds - Google Patents

Abstract

The present invention particularly provides methods for reducing lipids or reducing glucose, and compounds with an A-B ring structure of formula (I), in particular compounds which reduce lipids or reduce glucose.

<Formula I>

Wherein the dashed line between carbon 5 and 6 carbon, the dashed line between carbon 4 and R ² and the dashed line between carbon 11 and R ³ are independently of each other any doublet from each numbered carbon to the linked carbon Represents a bond and X is hydroxy or oxy linking carbon 8 and carbon 12 with Y.

Description

Glucose and Lipid Lowering Compounds

The present invention relates to certain compounds and compositions, as well as methods for alleviating hyperlipidemia and hyperglycemia.

This application claims priority to US Patent Application No. 09 / 190,507, filed November 12, 1998, and US Patent Application No. 09 / 197,352, filed November 20, 1998.

Harmful complications of hyperlipidemia are well known throughout the world for the high incidence of severe atherosclerosis, including heart attacks and heart attacks. Cardiovascular and cerebrovascular diseases are characterized by elevated blood levels of lipids and are slowly progressing symptoms that can be demonstrated by accumulated deposits of fatty substances in giant blood vessels for years or decades. Often not found. The disease exhibits its high morbidity and mortality only if sufficient blood vessel damage and accumulated thrombotic material are present to occlude or drop the main blood vessel and cause vascular occlusion.

In particular, the underlying factors within and beyond the controllability of an individual include genetic trends, comorbidities with diseases such as diabetes mellitus, or lifestyle factors such as diet, smoking and exercise. Affects. While genetic components have only recently become potential direct targets for therapeutic intervention, and lifestyle changes are difficult to achieve and often difficult to maintain, pharmacologically lowering blood lipid levels can reduce the incidence of cardiovascular and cerebrovascular diseases. Clinically successful in reducing. Its labels indicating elevated blood lipids and pathological conditions include free fatty acids, glycerol, triglycerides, cholesterol and low density lipoproteins. Alleviation or regulation of each of these components can have a positive effect on other abnormal blood analytes, including glucose and insulin.

Diabetes mellitus is a disease originating from other causes, including anabolic regulatory tissues as well as metabolic disorders associated with insulin producing tissues. In addition to diseases of hyperglycemia, some types of diseases are associated with abnormally elevated blood lipid levels. Thus, lowering blood levels of lipids and glucose is advantageous for the treatment of patients with diabetes mellitus. In type II diabetes, genetic mutations can cause certain subgroups of diseases (eg, adult diabetes or MODY in children), but most common types of diseases develop resistance to the action of insulin on insulin target tissues. Accompanying, predominantly associated with obesity of abdominal origin. In addition to elevated glucose levels, blood levels of triglycerides, cholesterol, and non-esterified fatty acids (NEFA) rise during fasting. In people with diabetes, cardiovascular disease, which is a major cause of morbidity and mortality, is likely to manifest. Resistance to insulin action develops not only in hyperglycemic states but also in syndrome X, an insulin resistant disease characterized by hyperinsulinemia and dyslipidemia (Reaven GM, Syndrome X, Clinical Diabetes, March / April 1994: 32-36). In this condition, normal glycemia can be maintained due to increased secretion of insulin. However, glucose intolerance coexists when insulin resistance cannot be maintained in the compensatory activity of the pancreas. Thus, apparent hyperglycemia or lipid abnormalities coupled with the normal glycemic syndrome X phenotype are thought to contribute to the severity of diabetic complications such as cardiovascular complications (Olefsky, JM, Current Approaches to the Management of Type 2 Diabetes: A Practical Monograph , National Diabetes Education Initiative, 1997).

Oral antihyperglycemic agents, such as sulfonylureas and biguanides, enhance the disposal of glucose and enhance the disposal of glucose by increasing the endogenous release of insulin from β-cells of the pancreas as in the case of sulfonylureas, or as in the case of biguanides. Alleviating the associated state of insulin resistance by mitigating promotes improvement of elevated lipid levels. Another approach to modulating elevated lipids and glucose involves the use of orally antihyperglycemic agents that act as widely as the thiazolidinedione family of agents that lower fatty acids and improve insulin resistance in insulin reactive tissues. Compound 4,7,8αH-Eudesma-5 (6), 11 (13) -diene-8,12-oxide, also known as helenin and allantolactone, causes rabbit hyperglycemia at high doses It has been reported to induce hypoglycemia at normal doses and to suppress hyperglycemia caused by food (1986, Handbook of Effective Ingredients of Medicinal Plants, Beijing, China).

Numerous different treatment strategies have been developed to treat hyperlipidemia and associated hyperglycemia of type II diabetes. The purpose of lowering blood lipids was to reduce cardiovascular morbidity and / or improve overall diabetes. Examples of classes of agents that act directly on plasma triglycerides and cholesterol levels include HMG-CoA reductase inhibitors, fibric acid and bile-salt resins. These classes are effective in lowering triglyceride and cholesterol levels, while having little effect on plasma fatty acids.

Nonesterified fatty acids in the lipid class appear to play a role in promoting diabetes and / or hyperlipidemic insulin resistance. Elevated free fatty acids result from excessive physical burden of fatty tissue in obese conditions, or unregulated cleavage of triglycerides in fatty tissue, major insulin target tissue, or both. In addition, elevated levels of free fatty acids have been shown to cause severe insulin resistance in muscle, major body tissues using glucose, and a direct effect on glucose delivery in the muscle is observed. In addition, fatty acids affect liver metabolism and increase hepatic glucose production. Elevated fatty acids also cause damaged α-cells that function by reducing the secretion of insulin from α-cells in response to glucose stimulation. Attempts to reduce fatty acid levels include the development of pyrogenic agents (agents) that are thought to function by stimulating brown fatty tissue to oxidize fatty acids and remove them from the blood. Alternatively, attempts have been made to develop fatty acid inhibitors by the enzyme carnitine palmitoyl transferase I (CPTI) to free fatty acids from their effects in the liver.

In particular, resistance to the action of insulin in adipose tissue appears to play a significant role in the elevation of fatty acids. Agents that inhibit the cleavage of fatty tissue of triglycerides are known as lipolytic inhibitors. It is the intention of the present invention to develop novel methods and agents for lowering blood lipids to treat hyperlipidemia diseases, or lowering blood glucose for treating hyperglycemia, diabetes and related diseases.

Summary of the Invention

The present invention relates to lipid lowering or glucose lowering methods and compounds having a ring A-B structure of the formula (I), in particular compounds lowering lipids or lowering glucose.

Wherein the dotted line between carbon 5 and 6 carbon, the dotted line between carbon 4 and R ² and the dotted line between carbon 11 and R ³ are independently of each other any double from each numbered carbon to the linked carbon Indicates binding.

The present invention particularly relates to methods of lowering the lipid level of a mammal to treat or prevent the progression of pathologies associated with elevated lipid levels; Compounds which inhibit the enzyme hormone-sensitive lipase and inhibit lipolysis; Agents and methods for the treatment of insulin resistance and syndrome X; Methods of lowering glucose levels in a mammal to treat or prevent the progression of pathologies associated with hyperglycemia; Provided are methods for treating impaired glucose tolerance associated with fasting or eating hyperglycemia, normal glycemia or insulin resistance, for example to delay or prevent the development of non-insulin dependent diabetes mellitus.

In accordance with the present invention, a method for reducing the lipid level of a mammal, comprising administering to a mammal (especially a hyperglycemic mammal) an effective amount of a composition characterized by comprising a compound of the formula A method of lowering glucose levels is provided.

The present invention comprises administering to a mammal a lipid lowering or glucose lowering effective amount of at least one compound of the AB ring structure of Formula (I) or a pharmaceutically acceptable salt thereof. In the formula I, the dotted line between carbon 5 and carbon 6, the dotted line between carbon 4 and R ² and the dotted line between carbon 11 and R ³ are each independently numbered carbon. Any double bond to carbon linked from R ¹ represents a hydrogen, hydroxy, lower alkanoyloxy or aroyloxy group wherein the aryl of aroyloxy is one or more lower alkyl, lower alkoxy, halo, trifluoro Romethyl, di (lower) alkylamino, hydroxy, nitro or C ₁ -C ₃ alkylenedioxy group), and X is hydroxy or Y Is an oxy linking carbon 8 and carbon 12 with Y being hydroxy, or an amino group which may be mono- or di-substituted with lower alkyl (where lower alkyl may be bonded to form a 5- or 6-membered ring) R ² and R ³ are independently of each other a lower alkyl or alkenyl group, lower alkyl of R ³ may be substituted on carbon linked to C11 with an amino group which may be mono- or di-substituted with lower alkyl, the lower alkyls being bonded To form a five or six membered ring, provided that when R ¹ is hydrogen, (a) X is hydroxy or (b) R ³ is substituted with amino. The invention also relates to a dosage pharmaceutical composition.

The invention also relates to the dotted line between carbon 5 and 6, the dotted line between carbon 4 and R ² and the dotted line between carbon 11 and R ³ independently of each other from each numbered carbon to the linked carbon. Represents a double bond, and R ¹ represents a hydrogen, hydroxy, lower alkanoyloxy or aroyloxy group, where aryl of aroyloxy is one or more lower alkyl, lower alkoxy, halo, trifluoromethyl, di (lower) Alkylamino, hydroxy, nitro or a C ₁ -C ₃ alkylenedioxy group), X is hydroxy or oxy which links carbon 8 and carbon 12 together with Y, and Y is hydroxy Or an amino group which may be mono- or disubstituted with lower alkyl, wherein lower alkyl may be bonded to form a 5- or 6-membered ring, and R ² and R ³ are independently lower alkyl or alkenyl groups, and R ³ , lower alkyl is lower And lower alkyl may be substituted to form a 5- or 6-membered ring, provided that (1) when X is hydroxy, R ¹ is hydrogen; And (2) when R ¹ is hydroxy, (a) X is hydroxy or (b) R ³ is substituted with amino, (3) R ² is methylene linked to C4 by a double bond and R ³ When methylene connected to C11 by this double bond and X is oxy, R ¹ relates to a compound of formula (I) (and its pharmaceutical composition) or a pharmaceutically acceptable salt thereof, which is not ethanoyloxy. Preferably, R ¹ is not propanoyloxy.

In one embodiment, the hydrogen substituents at positions 5, 7, 8 are in the alpha position relative to the AB ring structure. In another embodiment, X is hydroxy. In another embodiment, R ¹ is a lower alkanoyloxy group selected from the group consisting of, for example, acetoxy, propanoyloxy, isobutyryloxy and cyclopropanoyloxy. For example, the compound may comprise 2α-acetoxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-propanoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-cyclopropanoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-acetoxy-4,5,7,8,11αH-Eudesman-8,12-oxide; 2α-cyclopropanoyloxy-5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide. In another embodiment, R ¹ is an aroyloxy group such as peroyloxy and benzoyloxy, for example 2α-benzoyloxy-5,7,8αH-udesma-4 (15), 11 (13) -diene -8,12-oxide; 2α-perroyloxy-5,7,8αH-udesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-benzoyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide; Or 2α-benzoyloxy-5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide.

The compound may be administered in a lipid lowering effective amount or glucose lowering effective amount.

In one embodiment, (1) X is oxy, (2) R ¹ is lower alkanoyloxy or aroyloxy, R ² is methylene linked to C4 by a double bond and R ³ is linked to C11 by a double bond When linked methylene, R ¹ is not ethanoyl, and (3) R ² and R ³ are independently lower alkyl or lower alkenyl groups with limited unsaturation as linkages to carbon 4 or carbon 11 independently of one another. In another embodiment, (1) X is oxy, (2) R ¹ is lower alkanoyloxy or aroyloxy, and (3) R ² and R ³ are independently of each other to carbon 4 or carbon 11 Lower alkyl group or lower alkenyl group having limited unsaturation as the

Lower alkyl and alkenyl groups contain 1-6 carbons. Lower alkenyl groups are defined in relation to the framework structure of formula (I) such that the methylene group has its unsaturation, for example in connection with carbon 4 or carbon 11. In one embodiment, at least one or both of R ¹ and R ² are methyl or methylene (both bonded to a carbon 4 or 11 carbon with a double bond). The lower alkanoyloxy groups mentioned above contain 1 or 2 to 6 carbon atoms and are metanoyloxy, ethanoyloxy or acetoxy, propanoyloxy, butyryloxy, pentanoyloxy, hexanoyloxy, And corresponding branched chain isomers thereof. In the present application, among the alkanoyloxy groups, substituted and unsubstituted cycloalkyl groups such as cyclopropanoyloxy, cyclobutanoyloxy, cyclopentanoyloxy, cyclohexanoyloxy, 1-methylcyclohexanoyloxy, trans-4 -Isopropylcyclohexanoyloxy and the like. Aroyloxy groups as referred to herein are substituted and unsubstituted aromatic and heteroaromatic groups such as perroyloxy, benzoyloxy, 4-fluorobenzoyloxy, 2-chlorobenzoyloxy, 2-methoxybenzoyloxy, 3- Ethylbenzoyloxy, 4-ethylbenzoyloxy, 4-isopropylbenzoyloxy, 4-chlorobenzoyloxy, 3,5-tert-dibutyl-4-hydroxybenzoyloxy, 2-methoxy-5-chlorobenzoyloxy, 3-fluoro-4'-methoxybenzoyloxy, 2-trifluoromethylbenzoyloxy, 4-trifluoromethylbenzoyloxy, 2-naphthoyloxy, 2-thiophencarbonyloxy, 3-pyridinecarbonyl Oxy and 5-methyl-2'-pyrazinecarbonyloxy, including both 2α and 2β isomers, with the 2α isomer being preferred. Lower alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl and the corresponding branched chain isomers thereof. Preferably, the aryl group comprises a C ₆ -C ₁₀ aromatic group, or a heteroaromatic group having 5 to 10 ring atoms, preferably wherein at most two of the ring atoms are heteroatoms selected from nitrogen, oxygen or sulfur.

Lower alkyl groups mentioned above include methyl, ethyl, propyl, butyl, pentyl, hexyl and the corresponding branched chain isomers thereof. As noted above, the bond linking groups R ² and R ³ may be single or double bonds and the alkyl groups described herein may be bonded to carbon 4 and 11 respectively by single or double bonds.

Non-limiting examples of compounds in which R ¹ is hydrogen include 8β-hydroxy-4,7,8αH-Eudesma-5,11 (13) -diene-12-osan; 8β-hydroxy-5,7,8,11αH-Eudesme-4 (15) -ene-12-osan; 8β-hydroxy-4,5,7,8,11αH-Eudesman-12-osan; Salts such as the sodium salt of 2α-hydroxy-4 (15), 11 (13) udesadiene-12-osan. Non-limiting examples of compounds where R ¹ is hydroxy include 2α, 8β-dihydroxy-2β-4,5,7,8,11αH-udesmanman-12-osan; 2α, 8β-dihydroxy-4 (15), 11 (13) -udesadiadiene-12-osan; 2α-benzoyl-8β-hydroxy-2β, 4,5,7,8,11αH-udexaman-12-osan; 1- [2α-hydroxy-11,12-dihydro-5,7,8αH-Eudesme-4 (15) -ene-8,12-olidil] -pyrrolidine; And sodium or hydrochloride salts of 1- [2α-benzoyloxy-11,12-dihydro-5,7,8αH-Eudesme-4 (15) -ene-8,12-olidyl] -pyrrolidine. Salts.

Compounds wherein R ¹ is a lower alkanoyloxy group include 2α-acetoxy-5,7,8αH-udesma-4 (15), 11 (13) -diene-8,12-oxides; 2α-propanoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-isobutyryloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; α-cyclopropanoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-acetoxy-4,5,7,8,11αH-Eudesman-8,12-oxide; And 2α-cyclopropanoyloxy-5,7,8,11αH-udesma-4 (15) -ene-8,12-oxides. Compounds wherein R ¹ is an aroyloxy group include, for example, 2α-benzoyloxy-5,7,8αH-udesma-4 (15), 11 (13) -diene-8,12-oxide; And 2α-perroyloxy-5,7,8αH-udesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-benzoyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide; And 2α-benzoyloxy-5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide.

Hyperlipidemia When administered to a mammal, the compounds of the present invention have a positive effect on blood lipid levels. In addition, in vitro, the compounds specifically inhibit enzymes, hormone sensitive lipases (HSLs) and inhibit lipid degradation in whole cell models. In vivo, the compounds described herein inhibit the appearance of increased blood free fatty acids in normal mice fasted overnight. In the genetic model of obesity / type II diabetes associated with hyperlipidemia, the compound alleviates elevated blood lipid levels.

Compounds of the invention are prepared from synthetic starting materials or those isolated from natural sources. One specific starting material having a double bond between carbon 4 and 15 and between carbon 11 and 13, 2α-hydroxy-5,7,8αH-udesma-4 (15), 11 (13) -Diene-8,12-old can be synthesized by the procedure described by Tomioka et al. (1984, Tetrahedron Letters 25: 333-336), or by Herz et al. (1962, J. Org. Chem. 27: 905-910) can be isolated and purified from the plant Iva microcephala or other plants. Alkanoyloxy and aroyloxy derivatives of 2-hydroxy compounds such as 2α-hydroxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxides Prepared by synthetic procedures known to the skilled person, for example 2α-propanoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide and 2α-Benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide is 2α-hydroxy-5,7,8αH-Eudesme-4 ( 15), 11 (13) -diene-8,12-oxide, prepared by reaction with propanecarbonyl chloride and benzoyl chloride, respectively. In order to prepare a compound of the invention wherein R ² and R ³ and each ring carbon is a single bond, 2α-hydroxy-5,7,8αH-Eudesme-4 (15), 11 (13) -diene Dienes such as -8,12-oxides are first used, for example, according to the procedure of Hertz et al. (1962, J. Org. Chem. 27: 905-910), 2α-hydroxy-4,5,7,8,11αH. Reduced to Eudesman-8,12-oxide, followed by reaction with, for example, acyl chloride or benzoyl chloride, respectively 2α-acetoxy-4,5,7,8,11αH-Eudesman-8,12-ol Lead and 2α-benzoyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide. These compounds, in which only one of R ² or R ³ is double bonded to the ring, can be isolated or prepared synthetically or from natural sources according to synthetic procedures similar to those described above. For example, Compound 5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 4,7,8αH-Eudesma-5 (6), 11 (13) -diene-8,12-old and 5,7,8,11αH-Eudesme-4 (15) -en-8, 12-olide is commercially available from Sigma Chemical Co., St. Louis, MO, by silica gel chromatography (e.g., loaded with petroleum ether and eluting with increasing concentration of ethyl ether). Can be isolated from 80% isolehenin. 4,5,7,8,11αH-Eudesmann-8,12-Olyde reduces 4,7,8αH-Eudesma-5 (6), 11 (13) -diene-8,12-Old It can manufacture by doing.

Useful sources of compounds or starting materials include Bohlmann et al. (“New Sesquiterpene Lactones fom Inula Species,” 1978, Phytochem. 17: 1165-1172); Topcu et al. (“Cytotoxic and Antibacterial Sesquiterpenes from Inula Graveolens,” 1993, Phytochem. 33: 407-410); Ohta et al. ("Sequiterpene Constituents of Two Liverworts of Genus Diplophyllum, 1977, Tetrahedron 33: 617-628); Hertz et al. (1964," Constituents of Iva Species II, "J. Org. Chem. 29: 1022-1026) It is a plant isolate described by numerous authors.

Compounds in which R ³ incorporates amino, such as similar adducts described by Hejchman et al. (1995. 38: 3407-3410), can be synthesized by Michael addition reaction. The ring-opening compound can be synthesized by hydrolysis of the corresponding lactone.

Typical synthetic procedures are described in the examples below. Both 2α and 2β isomers are included, with 2α isomers being preferred. 2β isomers can be prepared from the corresponding 2α isomers using the Mitsunobu reaction (Mitsunobu et al., 1967, Bull. Chem. Soc. Japan 40: 935).

Representative compounds of the present invention include 2α-cyclohexanecarbonyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide, 2α-cyclopentanecarbonyloxy-4,5,7, 8,11αH-Eudesman-8,12-oxide, 2α-cyclobutanecarbonyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide, 2α- (2'- Naphthoyloxy) 5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2α- (1'-methylcyclohexanecarbonyloxy) -5,7,8αH- Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (4'-trifluoromethylbenzoyloxy) -4,5,7,8,11αH-Eudesmann -8,12-oxide, 2α- (2'-naphthoyloxy) -4,5,7,8,11αH-Eudesman-8,12-oxide, 2α-cyclohexanecarbonyloxy-5, 7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (3 ', 5'-di-tert-butyl-4'-hydroxybenzoyloxy) -5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2α- (2'-methoxy-5'-chlorobenzoyloxy) -4,5,7, 8,11αH-Eudesmann-8,12-oxide, 2α- (trans-4'-isopropylcyclohexanecarbonyloxy) -5, 7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α-cyclopentanecarbonyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (4'-ethylbenzoyloxy) -5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide , 2α- (3'-ethylbenzoyloxy) -5,7,8,11αH-udesme-4 (15) -ene-8,12-oxide, 2α-cyclobutanecarbonyloxy-5,7, 8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (2'-methoxybenzoyloxy) -5,7,8,11αH-Eudesme-4 (15) -en-8,12-oxide, 2α- (2'-trifluoromethylbenzoyloxy) -5,7,8,11αH-udesme-4 (15) -ene-8,12- Old, 2α- (3'-fluoro-4'-methoxybenzoyloxy) -5,7,8,11αH-udesme-4 (15) -ene-8,12-oxide, 2α- ( 2'-methoxy-5'-chlorobenzoyloxy) -5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2α- (2α- (2'-chloro Benzoyloxy) -5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2α- (4'-fluorobenzoyloxy) -5,7,8,11αH- Eudesme-4 (15) -en-8,12-old, 2α- (trans-4'-iso Lofilcyclohexanecarbonyloxy) -5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (1'-methylcyclohexanecarbonyloxy) -5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (4'-fluorobenzoyloxy) -4,5,7,8, 11αH-Eudesman-8,12-oxide, 2α- (2α- (2'-chlorobenzoyloxy) -4,5,7,8,11αH-Eudesman-8,12-oxide, 2α- (2'-methoxybenzoyloxy) -4,5,7,8,11αH-Eudesman-8,12-oxide, 2α- (3'-ethylbenzoyloxy) -4,5,7,8, 11αH-Eudesman-8,12-oxide, 2α- (4'-ethylbenzoyloxy) -4,5,7,8,11αH-Eudesman-8,12-oxide, 2α- (4 ' Isopropylbenzoyloxy) -4,5,7,8,11αH-Eudesmann-8,12-oxide, 2α- (4'-chlorobenzoyloxy) -4,5,7,8,11αH-oil Dessman-8,12-Old, 2α- (3 ', 5'-tert-dibutyl-4'-hydroxybenzoyloxy) -4,5,7,8,11αH-Eudesman-8,12 -Old, 2α- (2'-naphthoyloxy) -5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (3'- Fluoro-4'-methoxybenzoyloxy) -4,5,7,8,1 1αH-Eudesman-8,12-oxide, 2α- (2'-trifluoromethylbenzoyloxy) -4,5,7,8,11αH-Eudesman-8,12-oxide, 2α- (4'-isopropylbenzoyloxy) -5,7,8,11αH-udesme-4 (15) -ene-8,12-oxide, 2α- (4'-chlorobenzoyloxy) -5,7 , 8,11αH-Eudesme-4 (15) -en-8,12-oxide, 2α-cyclohexanecarbonyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -Diene-8,12-oxide, 2α-cyclopentanecarbonyloxy-5,7,8αH-eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- ( 4'-trifluoromethylbenzoyloxy) -5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2α-cyclobutanecarbonyloxy-5,7,8αH -Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (1'-methylcyclohexanecarbonyloxy) -4,5,7,8,11αH-Eudes Man-8,12-Old, 2α- (trans-4'-isopropylcyclohexanecarbonyloxy) -4,5,7,8,11αH-Eudesman-8,12-Old, 2α- ( 4'-fluorobenzoyloxy) -5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (2α- (2 ' -Chlorobenzoyloxy) -5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (2'-methoxybenzoyloxy) -5,7 , 8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (3'-ethylbenzoyloxy) -5,7,8αH-Eudesma-4 (15 ), 11 (13) -diene-8,12-oxide, 2α- (4'-ethylbenzoyloxy) -5,7,8αH-udesma-4 (15), 11 (13) -diene-8 , 12-oxide, 2α- (4'-isopropylbenzoyloxy) -5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- ( 4'-Chlorobenzoyloxy) -5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (3 ', 5'-tert-dibutyl -4'-hydroxybenzoyloxy) -5,7,8αH-udesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (2'-methoxy-5 ' -Chlorobenzoyloxy) -5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (3'-fluoro-4'-methoxybenzoyl Oxy) -5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α- (2'-trifluoromethylbenzoyloxy) -5,7, 8αH-Eudesma-4 (15), 11 (13) -diene-8,12-old, 2α- (4'-triple Oromethylbenzoyloxy) -5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α-benzoyloxy-4,5,7,8,11αH- Eudesman-8,12-Old, 2α-benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-Old, 2α-benzoyloxy- 5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2β-benzoyloxy-4,5,7,8,11αH-Eudesmann-8,12-ol Reed, 2β-benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2β-benzoyloxy-5,7,8,11αH-oil Desme-4 (15) -ene-8,12-oxide, 2α-cyclopropanoyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide, 2α-cycloprop Panoyloxy-5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2α-cyclopropanoyloxy-5,7,8αH-Eudesma-4 ( 15), 11 (13) -diene-8,12-oxide, 2β-cyclopropanoyloxy-4,5,7,8,11αH-Eudesmann-8,12-oxide, 2β-cycloprop Panoyloxy-5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2β-cyclopropanoyloxy-5,7,8αH-Eudesma-4 ( 15), 11 (1 3) -diene-8,12-oxide, 2α-perroyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α-perfluoro Iloxy-5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2α-perroyloxy-4,5,7,8,11αH-Eudesmann-8 , 12-Old, 2β-Perroyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-Old, 2β-Perroyloxy-5,7 , 8,11αH-Eudesme-4 (15) -en-8,12-oxide, 2β-perroyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide, 2α-isobutyryloxy-5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2α-isobutyryloxy-4,5,7,8,11αH- Eudesman-8,12-Old, 2α-Isobutyryloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-Old, 2β-iso Butyryloxy-5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2β-isobutyryloxy-4,5,7,8,11αH-Eudesmann -8,12-Old, 2β-Isobutyryloxy-5,7,8, αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α-propano Iloxy-4,5,7,8,11αH-Eudesmann-8,12-Old, 2α-propanoyl Ci-5,7,8,11αH-Eudesme-4 (15) -en-8,12-oxide, 2α-propanoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2β-propanoyloxy-4,5,7,8,11αH-Eudesmann-8,12-oxide, 2β-propanoyloxy-5 , 7,8,11αH-Eudesme-4 (15) -en-8,12-oxide, 2β-propanoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13 ) -Diene-8,12-oxide, 2α-acetoxy-5,7,8αH-eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2α-acetoxy- 5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide, 2α-acetoxy-4,5,7,8,11αH-Eudesmann-8,12-ol Reed, 2β-Acetoxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide, 2β-acetoxy-5,7,8,11αH-u Desme-4 (15) -ene-8,12-oxide, 2β-acetoxy-4,5,7,8,11αH-Eudesmann-8,12-oxide, 4,5,7,8 , 11αH-Eudesmann-8,12-Old, 4,7,8αH-Eudesma-5 (6), 11 (13) -diene-8,12-Old, 5,7,8αH-U Desma-4 (15), 11 (13) -diene-8,12-old, 5,7,8,11αH-Eudesme-4 (15) -en-8,12-old, 2α- Hydrock -4 (5), 11 (13) udedesadiene-12,8-oxide, 1- [sodium 2α-benzoyloxy-8β-hydroxy-11,13-dihydro-5,7,8,11αH -Eudesme-4 (13) ene-12-acetyl] -pyrrolidine hydrochloride, sodium 8β-hydroxy-4,5,7,8,11αH-Eudesman-12-oate, sodium 8β -Hydroxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-12-oate, sodium 8β-hydroxy-4,7,8αH-Eudesma-5 (6 ), 11 (13) -diene-12-oate, sodium 8β-hydroxy-5,7,8,11αH-Eudesme-4 (15) -ene-12-oate, sodium 2α, 8β-di Hydroxy-2β-4,5,7,8,11αH-Eudesman-12-Oate, Sodium 2α, 8β-dihydroxy-4 (15), 11 (13) -Eudesadiene-12- Oate, sodium 2α-benzoyl-8β-hydroxy-2β, 4,5,7,8,11αH-Eudesman-12-oate, 8β-hydroxy-4,7,8αH-udesama-5, 11 (13) -diene-12-oxane, 8β-hydroxy-5,7,8,11αH-Eudesam-4 (15) -ene-12-acid, 8β-hydroxy-4,5,7,8 , 11αH-Eudesman-12-Osan, 2α, 8β-dihydroxy-2β-4,5,7,8,11αH-Eudesaman-12- Acid, 2α, 8β-dihydroxy-4 (15), 11 (13) -udesadiadi-12-osan, 2α-benzoyl-8β-hydroxy-2β, 4,5,7,8,11αH-oil Desaman-12-osan, 1- [2α-hydroxy-11,12-dihydro-5,7,8αH-Eudesme-4 (15) -ene-8,12-olidyl] -pyrrolidine ;

1- [2α-benzoyloxy-11,12-dihydro-5,7,8αH-Eudesme-4 (15) -ene-8,12-olidil] -pyrrolidine.

After synthesis, the compounds of the present invention are isolated and purified by established methods to produce pharmaceutically acceptable substances for administration to mammals.

The compounds used in the methods of the invention may preferably be administered as medicaments, ie pharmaceutical compositions, preferably administered. Preferred oral and intraperitoneal dosage forms of the compound are preferred.

Pharmaceutical compositions for use in the methods of the invention for administration to animals and humans are characterized in that they comprise a compound of the invention incorporating a pharmaceutical carrier and excipients. The medicament may be in the form of a tablet, dragee, capsule, pill, ampoule or suppository, characterized in that it comprises a compound of the invention.

As used herein, "pharmaceutical" means a physically discrete aggregated portion suitable for drug administration. As used herein, a "dose in the form of a dosage unit" refers to the active compound of the invention, each associated with a carrier and / or encapsulated in an envelope, of several times or less (four times) or dozens (less than four times) of a daily or daily dosage. Hereinafter) means physically discrete aggregated units suitable for pharmaceutical administration containing a quantity. Whether the medicament will contain a daily dose or, for example, 1/2, 1/3 or 1/4 of the daily dose will depend on whether the medicament is once a day or, for example, twice, three or four times a day. It will depend on whether it will be administered.

Advantageously, the composition is formulated as a dosage unit, each unit adapted to provide a fixed dosage of the active ingredient. Tablets, coated tablets, capsules, ampoules and suppositories are examples of preferred dosage forms according to the present invention. It is only required that the active ingredient constitutes an effective amount. In other words, a suitable effective dosage will be consistent with the dosage form used in single or multiple unit dosages. Of course, the exact daily dose as well as the individual dose will be determined according to standard medical principles under the direction of a physician or veterinarian.

The compounds of the invention can also be administered as suspensions, solutions and emulsions, syrups, granules or powders of the active ingredients in aqueous or non-aqueous diluents.

Diluents that can be used in pharmaceutical compositions (eg granules) containing the active compound adapted to form tablets, dragees, capsules and pills include (a) fillers and extenders such as starch, sugars, mannitol and silicic acid; (b) binders such as carboxymethyl cellulose and other cellulose derivatives, alginates, gelatin and polyvinyl pyrrolidone; (c) humectants such as glycerol; (d) disintegrants such as agar-agar, calcium carbonate and sodium bicarbonate; (e) agents for delaying dissolution, such as paraffin; (f) resorption accelerators, such as quaternary ammonium compounds; (g) surface active agents such as cetyl alcohol, glycerol monostearate; (h) adsorptive carriers such as kaolin and bentonite; (i) lubricants such as, but not limited to, talc, calcium and magnesium stearate and solid polyethylene glycols.

Tablets, dragees, capsules and pills characterized by comprising the active compound may comprise conventional coatings, shells and protective matrices which may contain opacifying agents. They may be configured to release the active ingredient, possibly only at a specific site of the intestine or preferably at that particular site for a period of time. Coatings, sheaths and protective matrices can be made, for example, of polymeric materials or waxes. The compounds of the present invention can also be prepared by microencapsulation with one or several of the diluents. Diluents used in pharmaceutical compositions adapted to be formed into suppositories are for example polyethylene glycols and fats (e.g. cocoa oil and higher esters such as C ₁₄ alcohols with C ₁₆ fatty acids) or mixtures of these diluents. General water soluble diluents.

Pharmaceutical compositions that are solutions and emulsions may contain conventional diluents such as, for example, solvents, solubilizers, and emulsifiers. Specific and non-limiting examples of such diluents are water, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil Fatty acid esters of peanut oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitol, or mixtures thereof. For parenteral administration, solutions and suspensions, such as water or arachis oil, should be sterile and, if appropriate, blood isotonic. Pharmaceutical compositions that are suspensions include common diluents, such as liquid diluents, for example water, ethyl alcohol, propylene glycol, surface active agents (eg, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters), microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth or mixtures thereof.

Pharmaceutical compositions may also contain flavoring and flavoring additives (eg peppermint oil and eucalyptus oil), as well as colorants and preservatives, sweeteners (eg saccharin and aspartame). Pharmaceutical compositions will generally contain from 0.5 to 90% by weight of the active ingredient of the total composition. In addition to the compounds of the present invention, pharmaceutical compositions and medicaments may also contain other pharmaceutically active compounds. Any diluent in the medicament of the invention may be any of those mentioned above with respect to the pharmaceutical composition. Such agents may include solvents having a molecular weight of less than 200 as the sole diluent. Compounds of the invention will be administered orally, parenterally (eg intramuscular, intraperitoneal, subcutaneous, transdermal or intravenous), rectally or topically, preferably orally or parenterally, in particular sublingually or intravenously. I think.

For example, a dosage of 0.5 to 500 mg per kg of body weight may vary depending on the nature and body weight of the human or animal being treated, the individual response of the treatment subject, the type of formulation of the active ingredient to be administered, the mode of administration, the progression of the disease or the interval at which it is administered. It will be a function. Thus, in some cases it will be sufficient to use less than the minimum dose, but in other cases the upper limit will have to be exceeded to obtain the desired result. If higher doses are administered, it may be wise to divide into several doses throughout the day.

While not intending to be bound by any particular theory, it is believed that by inhibiting the activity of hormone sensitive lipases, the compounds of the present invention interfere with the lipolysis of stored triglycerides and their metabolism to fatty acids and glycerol. As a result of metabolism, elevated glucose levels can be reduced. Metabolism of fat provides three times more calorie energy than carbohydrates or proteins. As such, fats are fuel sources of limited availability due to their relatively abundance. In the liver, free fatty acids undergo α-oxidation, which simultaneously inhibits glycolysis by the negative five-bag mechanism while simultaneously promoting your life. In addition, the released glycerol is used as a three-carbon substance in your life, contributing to increased levels of hepatic glucose production. Under normal metabolic regulatory environment, enhanced lipolysis of storage triglycerides in adipose tissue occurs after an extended fasting period and is rapidly inhibited after meal digestion by the action of insulin. However, in the insulin resistant state of type II diabetes, this inhibition of lipid degradation by insulin is lost even after meals. In this situation, the liver remains constant in your life and glucose delivery to muscle tissue is reduced, resulting in overt hyperglycemia.

The antilipidic effect of the compounds of the present invention in adipose tissue results in a decrease in plasma free fatty acid and glycerol concentrations. In addition, the conversion from your life to glycolysis in the liver can be achieved due to the reduced use of fatty acids as a fuel source, and the resulting drop in plasma glucose levels is reset again as the first energy source. The degradation of free fatty acids further affects the functioning of the pancreas. In β-cells, the elevation of free fatty acids is evidenced by compounds that exhibit NEFA-lowering properties that prevent and restore proper β-cell function, as demonstrated by Unger (1995, Diabetes 44: 863-870). Causes damage to the secretion mechanism. Thus, the compounds of the present invention, in addition to their beneficial effects on blood lipids, achieve lowered or normalized elevated blood glucose and insulin levels, and alleviated muscle insulin resistance.

Combining the results of enzyme inhibition studies, lipolysis assays, and animal studies described herein, it can be seen that the compounds of the present invention have unique mechanisms of action and will not exhibit the toxic cell modification effects observed in conventional thiazolidinediones. . These are thus very useful and preferred therapeutic agents in various disease states where lowering blood glucose levels and / or inhibition of lipolysis is required for the treatment of patients. Such disease states include diabetes, and in particular adult diabetes, or type II diabetes, insulin resistance associated with dyslipidemia, and hyperglycemia, referred to as syndrome X, hypertension and atherosclerosis.

Enzyme hormone-sensitive lipase (HSL) is distinguished from other endogenous lipase enzyme systems in that its activity is governed by the action of various steroidal and nonsteroidal hormones, and secretory neurotransmitters through the receptor mediated second messenger pathway. Examples of such substances include insulin, corticosteroids and norepinephrine. The first location of the hormone sensitive lipase is in adipose tissue, where it is involved in the equal regulation of lipid metabolism. As noted above, the activity of the enzyme is responsible for the mobilization of stored triglycerides to provide a fuel source upon fasting. Recent expression of genes for mammalian hormone sensitive lipases in transfected bacterial vectors provides a means for the isolation of pure hormone sensitive lipase proteins. This led to the development of in vitro assays that assess the direct effect of potential therapeutic agents on the activity of the enzyme itself without disturbing other signaling and regulatory entities found in the whole cell line.

A suitable model for assessing lipid cell function in tissue culture conditions is transformed NIH-3T3 cells. Incubation of 3T3 cell lines with dexamethasone and 3-isobutyl-1-methylxanthine (IBMX) prevented the differentiation of fibroblastic blast cells into mature adipocytes capable of insulin-regulated liposynthesis and norepinephrine-mediated lipolysis. cause. Thus, inhibition of lipolytic activity can be assessed for latent agents in whole cells and in permeated conditions. This type of assessment is used to correlate with the effectiveness of potential therapeutics evaluated in purified enzymatic systems in an appropriate and intact biological environment.

Under normal metabolic control, cleavage of storage triglycerides in the mammalian fat tissue is tightly regulated by the hormone insulin. When insulin levels are as high as after release of β-cells from the pancreas in response to glucose's high blood blood levels, the activity of hormone sensitive lipase is inhibited. When glucose levels decrease, insulin levels decrease in a parallel manner. At some point during the extended fasting period, insulin levels are reduced to levels below that required for inhibition of hormone sensitive lipase. As a result, the catalytic activity of the enzyme is enhanced to cause release of the unesterified free fatty acids (NEFA), and glycerol from the storage triglyceride reservoirs.

Thus, normal animals without diabetes can be used for the determination of hormone sensitive lipase inhibition by administering test compounds prior to the start of the fasting period with high insulin levels. Decrease in the level of plasma NEFA after an extended fasting period of 18 hours would suggest inhibiting the recruitment of triglyceride reservoirs from the lipid site via the hormone sensitive lipase pathway.

In type II diabetes, the insulin regulatory mechanisms for lipolysis do not function properly. Due to the state of insulin resistance, in certain cases, despite elevated insulin levels over the normal range, the inhibition of hormone sensitive lipase activity is severely reduced, resulting in enhanced lipolytic activity against stored triglycerides, resulting in highly elevated fasting plasma NEFA Cause levels. In addition, recent experimental evidence suggests that a transient state of insulin resistance in normal individuals without diabetes can be achieved following direct injection of NEFA into the circulation. This data will support the pathological role for elevated plasma NEFA in type II diabetes.

To date, the exact etiology of the development of adult type II diabetes is unknown. However, 70% of these patient populations correlate with significant obesity because they can be classified as obese in the general criteria set by the American Diabetes Association, National Institutes of Health guideline. Thus, non-human mammals with similar pathophysiological profiles are suitable as models for evaluating anti-dyslipidemia and antidiabetic properties of potential agents.

Animal models showing these overall characteristics include ob / ob and KK / Ay strains of mice, and Zucker fa / fa (ZDF) rats. ob / ob mice begin to develop a diabetic profile at 4 weeks of age. Plasma glucose levels rise until 9-10 weeks, when the levels begin to stagnate. In addition to hyperglycemia, ob / ob mice show increased levels of NEFA, triglyceride production and insulin resistance. Moreover, parallel increases in plasma insulin levels appear as a result of hyperglycemia. When plasma insulin levels peaked, improvement in hyperglycemia is seen. However, the amount of insulin present is not sufficient to fully normalize the insulin resistance state during the oral glucose tolerance test.

Overall, KK / Ay is similar to ob / ob except for some of the differences mentioned. The onset of the Type II profile occurs much later, approximately after 3 months and 4 months of age, and lasts for 1 year. Both mouse models show hyperinsulinemia, hyperglycemia, insulin resistance, and dyslipidemia similar to those seen in clinical settings.

ZDF rats are also similar to ob / ob and KK / Ay mice in all general attributes of type II like profiles with one additional exception. In contrast to mouse models that show severe hyperinsulinemia, ZDF rats show a progressive decrease in their ability to secrete insulin from α-cells starting at 10 weeks of age, resulting in a state of hypoinsulinemia. Thus, these symptoms are similar in profile to subtypes of Type II diabetic patients who do not respond to certain classes of pharmacological agents, such as sulfonylureas, and may require insulin therapy.

The present invention may be better understood with reference to the following non-limiting examples provided as examples of the present invention. The following examples are provided to more fully illustrate preferred embodiments of the invention. However, they should not be construed as limiting the broad scope of the invention in any way.

Example 1 Preparation of Intermediates and Compounds

Isolation and Purification of 2α-hydroxy-5,7,8αH-Eudesme-4 (15), 11 (13) -diene-8,12-oxides: These compounds are intermediates in the preparation of the various compounds of the present invention. Used as Method reported by Hertz et al. Of 2α-hydroxy-5,7,8αH-Eudesme-4 (15), 11 (13) -diene-8,12-oxide (J. Org. Chem., 27: 905-910, 1962) was isolated from Iva microcephala Nutt. The soft stems of the heading, leaves and plants were dried, ground and extracted with a Soxhlet extractor with stirring for 5 hours with a mixture of petroleum ether and ether (2: 1 v / v). Crude 2-hydroxy-5,7,8α-Eudesme-4 (15), 11 (13) -diene-8,12-oxide began to precipitate in 2 hours. The extract was cooled overnight to room temperature, then filtered and washed with the same solvent mixture to afford crude 2-hydroxy-5,7,8α-Eudesme-4 (15), 11 (13) -diene-8,12- An oled was obtained (2.93% yield). Crude 2-hydroxy-5,7,8α-Eudesme-4 (15), 11 (13) -diene-8,12-oxide is crystallized from a mixture of CH ₂ Cl ₂ , ether and petroleum ether to give 2 -Hydroxy-5,7,8α-Eudesme-4 (15), 11 (13) -diene-8,12-oxide was obtained. Melting point: 127-129 ° C., yield: 2.86%,

Preparation of 2α-Benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide: The following scheme was used.

2α-hydroxy-udesma-4 (15), 11 (13) -diene-8,12-oxide (4.0 g, 16.1 mmol) is dissolved in CH ₂ Cl ₂ (13 ml) and 0 for 2 hours. Excess benzoyl chloride (4.0 g, 28.4 mmol) and pyridine (3.1 ml) were added with stirring at &lt; RTI ID = 0.0 &gt; C, &lt; / RTI &gt; and stirring continued overnight at room temperature. The reaction mixture was poured into ice water with stirring. The precipitate was collected by filtration and washed with ice water. The solid was dissolved in CH ₂ Cl ₂ (30 ml) and dried over anhydrous Na ₂ SO ₄ . The CH ₂ Cl ₂ solution was evaporated in vacuo to afford the crude product (7.0 g) which was purified by silica gel column chromatography eluted with CH ₂ Cl ₂ and 1% MeOH—CH ₂ Cl ₂ continuously. The product (5.08 g) was crystallized from a mixture of CH ₂ Cl ₂ , ether and petroleum ether to give 2α-benzoyloxy-5,7,8αH-udesma-4 (15), 11 (13) -diene-8,12 Yield (4.5 g, 79.3%) was obtained. Melting point: 127-129 ° C., purity 99.9% measured by HPLC,

Analyzes: C ₂₂ H ₂₄ C ₄ Theory: C% 74.90%, H 6.81%, Found: C% 75.06%, H% 6.95%

Preparation of 2α-hydroxy-4,5,7,8,11αH-Eudesman-8,12-oxide: 2α-hydroxy-4,5,7,8,11αH-Eudesman-8,12 -Olives were used as intermediates in the preparation of certain compounds of the invention. 2α-hydroxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide (4.0 g) in methanol (150 ml) containing acetic acid (1.5 ml) To the solution was added platinum oxide (0.12 g). The reaction mixture was hydrogenated in a Parr shaker at 2.72 atm (40 psi) at room temperature for 17 hours. The reaction mixture was filtered and concentrated in vacuo. The residue was washed with ice water to yield white powder product, 2α-hydroxy-4,5,7,8,11αH-Eudesman-8,12-oxide (3.36 g, 82.7%). Melting point: 154-155 ° C.

Preparation of 2α-benzoyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide: 2α-benzoyloxy-4,5,7,8,11αH-Eudesman-8,12 -Old was prepared according to the following scheme.

2α-hydroxy-4,5,7,8,11αH-Eudesman-8,12-oxide (3.36 g, 13.3 mmol) is dissolved in CH ₂ Cl ₂ (15 ml) and kept at 0 ° C. for 2 hours. Excess benzoyl chloride (4.0 g, 28.4 mmol) and pyridine (7.6 ml) were added with stirring, followed by continued stirring at room temperature overnight. The reaction solution was poured into ice water (700 ml) with evaporation and stirring. The precipitate was collected by filtration and washed with ice water. The solid was dissolved in CH ₂ Cl ₂ (50 ml) and dried over anhydrous Na ₂ SO ₄ . The CH ₂ Cl ₂ solution was evaporated in vacuo to afford crude product (7.51 g). The product was purified by silica gel column chromatography (eluate CH ₂ Cl ₂ ). The product was recrystallized from a mixture of CH ₂ Cl ₂ , ether and petroleum ether to give pure 2α-benzoyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide (3.38 g, 70.8%). Obtained. Melting point: 181-183 ° C. Purity 99.1% as determined by HPLC.

Analyzes: C ₂₂ H ₂₈ O ₄ Theory: C% 74.15%, H% 7.86%, Found: C% 74.17%, H% 8.00%

Compound 2α-Acetoxy-4,5,7,8,11αH-Eudesme-8,12-oxide was prepared according to the following procedure.

2α-cyclopropanoyloxy-5,7,8,11αH-Eudesman-4 (15) -ene-8,12-oxide and 2α-benzoyloxy-5,7,8,11αH-udedesme Preparation of -4 (15) -ene-8,12-oxides: These compounds are 2α-benzoyloxy-5,7,8αH-udesma-4 (15), 11 (13) -diene-8,12 2α-hydroxy-5,7,8,11αH-udesme-4 (15) -ene- using cyclopropanecarbonyl chloride and benzoyl chloride, respectively, according to the procedures described herein for the preparation of the oxides. Prepared from 8,12-oxides.

5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 4,7,8αH-Eudesma-5 (6), 11 (13) -diene-8,12-oxide; And Isolation and Purification of 5,7,8,11αH-Eudesme-4 (15) -en-8,12-oxides: Crude 5,7,8αH-Eudesma-4 (15), 11 (13 A commercially available product of) -diene-8,12-oxide, isohelenine (Sigma, Cat. 1-1639, purity 80%) was obtained. To purify 5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxides, petroleum ether with increasing ether amount was used as eluent (20.0 g). Silica gel chromatography gave 60 fractions of 100-150 ml each. Based on the similarity of TLC, the fractions were combined to yield three main fractions and further purified by rechromatography on a silica gel column with the same eluent. Fraction 2-4 (20.6%) containing 4,7,8αH-Eudesma-5 (6), 11 (13) -diene-8,12-oxide, 5,7,8αH-Eudesma Fractions 11-19 (57.5%) and -5,7,8,11αH-udesma-4 (15) -diene- containing -4 (15), 11 (13) -ene-8,12-oxide Fraction 41-51 (13.0%) containing 8,12-oxide. The structures of these compounds were inferred by high field NMR, including mass spectrometry, and 2D NMR studies.

4,7,8αH-Eudesma-5 (6), 11 (13) -diene-8,12-oxide: columnar, melting point (from a mixture of CH ₂ Cl ₂ , Et ₂ O and petroleum ether) : 73-75 ° C., purity 99.8% as determined by HPLC (Conditions: Column-Dynamax C-18, Detector-UV 203/210 nm, Flow rate-1.6 ml / min, Mobile phase-65% acetonitrile).

5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide: needle, melting point (from a mixture of CH ₂ Cl ₂ , Et ₂ O and petroleum ether) : 108-110 ° C., purity 99.8% as determined by HPLC (Conditions: column-Dynamax C-18, detector-UV 203/210 nm, flow rate-1.6 ml / min, mobile phase-65% ACN).

5,7,8,11αH-Eudesma-4 (15) -ene-8,12-oxide: needle-like (from mixture of CH ₂ Cl ₂ , Et ₂ O and petroleum ether) Melting Point: 171- Purity 99.4% as determined by HPLC (conditions: column-Dynamax C-18, detector-UV 203/210 nm, flow rate-1.6 ml / min, mobile phase-65% ACN).

Preparation of 4,5,7,8,11αH-Eudesman-8,12-Old: 5,7,8αH-Eudesma-4 in methanol (135 ml) containing acetic acid (1.0 ml) (15 To the solution, 11 (13) -diene-8,12-oxide (5.0 g, 0.021 mol) was added platinum (IV) oxide (0.12 g). The reaction mixture was hydrogenated overnight at 2.72 atm (40 psi) at room temperature. The reaction mixture was filtered and concentrated in vacuo. The residue was dissolved in methylene chloride (100 ml) and dried over anhydrous sodium sulfate. The methylene chloride solution was evaporated to give 4.95 g of crude product. The product was recrystallized from a mixture of methylene chloride and petroleum ether to give 4.52 g of 4,5,7,8,11αH-Eudesman-8,12-oxide. Melting point: 144-146 ° C., yield 88.9%.

Example 2A Additional Compounds

In a manner similar to that described in Example 1 above, the following additional compounds were prepared.

2α-Acetoxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide (melting point: 125-127 ° C.), 2α-isobutyryloxy-5 , 7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide (melting point: 76-78 ° C.), 2α-propanoyloxy-5,7,8αH-oil Desma-4 (15), 11 (13) -diene-8,12-oxide (melting point: 98-100 ° C.), 2α-perroyloxy-5,7,8αH-udesma-4 (15) , 11 (13) -diene-8,12-oxide (melting point: 98-100 ° C.), 2α-cyclopropanoyloxy-5,7,8αH-udesma-4 (15), 11 (13) -Diene-8,12-oxide (melting point: 160-163 ° C), 2α-acetoxy-4,5,7,8,11αH-Eudesman-8,12-oxide (melting point: 183-185 ° C) ), 2α-cyclopropanoyloxy-5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide (melting point: 157-160 ° C.), 2α-benzoyloxy-5 , 7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide (melting point: 150-153 ° C.).

Example 2B Additional Example

Preparation of 1- [2α-hydroxy-11,12-dihydro-5,7,8αH-Eudesme-4 (15) -ene-8,12-olidil] -pyrrolidine: 1- [2α Hydroxy-11,12-dihydro-5,7,8αH-udesme-4 (15) -ene-8,12-olidyl] -pyrrolidine described by Michael Hedgeman et al. (1995, 38: 3407-3410). Ivalin (2α-hydroxy-4 (15), 11 (13) udesadiene-12,8-oxide) (1.15 mmol) was dissolved in THF (5 ml). The solution was cooled in the refrigerator for 15 minutes. Pyrrolidine (3 mmol) in THF (5 ml) was added to a solution of invalin. The mixture was stored in a refrigerator at 5 ° C. for 20 periods. The solvent was evaporated. The residue was dissolved in ether and filtered through a pad of silica gel. The solvent was evaporated and the residue was placed under vacuum for 24 hours to remove traces of pyrrolidine. The product was crystallized from a mixture of ethyl acetate and hexanes in about 55% yield. Melting point: 121-124 ° C.

In the same way, 1- [11,12-dihydro-5,7,8αH-Eudesme-4 (15) -ene-8,12-olidil] -pyrrolidine isohelenine (5,7 , 8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide) and synthesizes 1- [2α-benzoyloxy-11,12-dihydro-5,7,8αH -Eudesme-4 (15) -ene-8,12-olidil] -pyrrolidine to 2α-benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene Synthesized from -8,12-oxide.

Example 2C Additional Example

Preparation of the ring-opening structure: The starting compound (8 mol) was added to a solution of NaOH (8 mol) in 90% EtOH (10-20 ml if necessary) and the mixture was stirred at 60-90 ° C. for 2-3 hours.

Starting compound product 4,5,7,8,11αH-Eudesmann-8,12-Old 8β-hydroxy-4,5,7,8,11αH-Eudesman-12-osan 4,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-old 8β-hydroxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-12-osan 4,7,8αH-Eudesma-5 (6), 11 (13) -diene-8,12-old 8β-hydroxy-4,7,8αH-udesama-5 (6), 11 (13) -diene-12-osan 5,7,8,11αH-Eudesme-4 (15) -en-8,12-old 8β-hydroxy-5,7,8,11αH-Eudesam-4 (15) -ene-12-osan 2α-hydroxy-4,5,7,8,11αH-Eudesmann-8,12-old 2α, 8β-dihydroxy-2β-4,5,7,8,11αH-Udesaman-12-osan 2α-hydroxy-5,7,8αH-Eudesme-4 (15), 11 (13) -diene-8,12-old 2α, 8β-dihydroxy-4 (15), 11 (13) -udesamadiene-12-osan 2α-benzoyloxy-4,5,7,8,11αH-Eudesmann-8,12-oxide 2α-benzoyl-8β-hydroxy-2β, 4,5,7,8,11αH-udexaman-12-osan

The solvent was evaporated at reduced pressure and the product carboxylate salt was recrystallized from CH ₃ CN and EtOH or MeOH and ethyl acetate.

Example 3 Dosage Form

The compounds of the present invention can be formulated for oral pharmaceutical administration to patients in need of blood glucose lowering as follows.

ingredient mg / tablet Compound of the Invention 50 Starch 50 Mannitol 75 Magnesium stearate 2 Stearic acid 5

The compound, a portion of starch and lactose were combined and wet granulated with starch paste. The wet granulation was placed in a tray and dried overnight at 45 ° C. The dried granulate was ground in a grinder to approximately 20 mesh particle size. Magnesium stearate, stearic acid and the remaining starch were added and the entire mixture was blended and then pressed on a suitable tablet press. The tablets were compressed to a weight of 232 mg using a 0.873 cm (11/32 inch) punch of 4 kg hardness. These tablets will disintegrate within 1/2 hour according to the method described in USP XVI.

Example 4 Hormone Sensitive Lipase

The potential to directly inhibit the activity of hormone sensitive lipase was measured in an in vitro purified enzyme system. The recombinant hormone sensitive lipase protein isolated from the bacterial expression vector is preincubated with a compound of the invention (eg 100 μM), followed by addition of the emulsified triolein phosphatidylcholine / phosphatidylinositol (PC / PI) material. The reaction was initiated. At various time points, an aliquot of the reaction was taken and the glycerol content was used as an indicator of enzyme activity. Pharmacological specificity was measured by culturing the test compound with other mammalian and non-mammalian lipase enzyme systems to assess inhibition of activity. Evaluation was performed using triolein concentrations from 42 μM to 835 μM.

Example 5 Inhibition of Hormone Sensitive Lipase

In a manner similar to that described in Example 4 above, 2α-benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide at 100 μM The ability to inhibit the activity of hormone sensitive lipase against cholesterol oleate PC / PI was evaluated. Evaluation was performed using cholesterol oleate at a concentration of 12.5 μM to 225 μM.

Example 6 Hormone Sensitive Lipase and 3T3-L1 Hormone Sensitive Lipase

In a manner analogous to that described in Example 4 above, 2α-benzoyloxy-4,5,7,8,11αH-Eudesman-8,12-olith at 100 μM 3T3 on glycerol stabilized triolein The ability to inhibit the activity of hormone sensitive lipase and recombinant hormone sensitive lipase derived from the preparation of cell lysates was evaluated. The results are shown in Table 1 below.

Triolein concentration (μM) % Suppression Refined HSL 3T3-L1 melt 30 77 63 67 78 56 100 82 51 201 64 41 400 50 38 601 40 30 800 18 18 1000 27 15

Example 7 Additional Enzyme Evaluation

To demonstrate the specificity of the compounds of the present invention for hormone sensitive lipases, 2α-benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxides Inhibition of Candida rugosa lipase against emulsified triolein substance PC / PI was evaluated. As can be seen in Table 2, 2α-benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide did not inhibit the enzyme activity.

Triolein substance, μM Candida Lipase (DMSO) sem Candida Lipase + Test Compound sem % Suppression 42 5.40 0.44 3.21 0.12 40 83.5 9.66 0.51 11.59 0.59 -12% 167 14.98 0.05 14.14 0.65 6.3 334 25.19 0.12 22.68 0.75 3.6 501 59.12 1.56 60.13 2.85 -1.7 668 91.67 14.97 93.59 11.56 -1.1 835 94.13 2.12 89.71 1.98 1.0

Example 8 Additional Enzyme Evaluation

The specificity of the compounds of the invention is catalyzed by the hydrolysis of glycerol stabilized triolein material with 2α-benzoyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide at 100 μM. It was further demonstrated by evaluating the inhibition of hepatic lipase and lipoprotein lipase. The results are shown in Table 3.

Triolein substance, μM Pure soy lipase Lipoprotein lipase (DMSO) sem + Test compound sem (DMSO) sem + Test compound sem 30 0.11 0.02 0.08 0.03 0.6 0.1 0.7 0.1 67 0.20 0.03 0.37 0.08 1.7 1.7 1.8 0.2 100 1.35 0.17 0.69 0.05 - - - - 134 - - - - 4.8 4.8 4.9 0.3 201 6.11 0.14 6.36 0.20 - - - - 264 - - - - 9.6 9.6 10.3 0.8 400 29.20 3.20 30.80 3.06 - - - - 1000 134.60 2.60 151.60 2.09 65.2 65.2 67.4 1.2

As can be seen from Table 5, compound 2α-benzoyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide failed to significantly inhibit the activity of the two enzymes evaluated.

Example 9 Effect of Compounds of the Invention on Lipid Degradation

Any in vitro assay for lipid degradation was performed using 3T3-L1 cells. In this assay protocol, 3T3-L1 adipocytes were perfused with Digitonin and then precultured with the compounds of the present invention. The hormone sensitive lipase was then activated with cAMP, causing the hydrolysis of triglycerides to free fatty acids and glycerol. Free glycerol in the supernatant was quantified by fluorescence analysis performed on a Microfluor B flat bottom plate.

The protocol of the lipolysis assay was as follows: 3T3-L1 adipocytes were used 14 days after the onset of differentiation. The medium was carefully removed from the fat cells. The cells were then washed gently twice with 300 μl of PBS. The cells were incubated with 0.2 ml of a permeable buffer consisting of 10 mM sodium phosphate, 150 mM sodium chloride, pH 7.4, 1% fatty acid free BSA and digitotonin (10 mg / ml). After 30 min incubation at 37 ° C., additional 0.2 ml of permeable buffer containing test compounds were added and cells were further incubated for 30 min. The buffer was then completely removed and contained 200 μl permeable buffer containing test compound and N ⁶ -benzoyloxy-cAMP (0.6 mM), 8-thimethyl-cAMP (0.6 mM), 2 mM ATP and 4 mM MgCl ₂ . Was replaced with 200 μl permeable buffer. cAMP was added to activate cAMP dependent protein kinase, followed by phosphorylate to activate hormone sensitive lipase. The cells were then incubated at 37 ° C. for 30-60 minutes. 350 μl aliquots of buffer were removed and mixed with 100 μl of 10% charcoal solution. The suspension was centrifuged (300 RPM, 20 minutes) and 50 μl of supernatant was transferred to a 96 well flat bottom plate (microfluor B) and frozen at −20 ° C. until analyzed for glycerol according to the following procedure.

Glycerol Analysis: The amount of glycerol produced during lipolysis was measured according to the following two step reaction.

The reaction buffer consisted of kinase buffer, pH 7.0 (2 mM MgCl ₂ , 100 mM triethanolamine, 2 mg / ml fatty acid free BSA), 130 μM ATP, 2 units / ml of riser kinase and 1 unit / ml of glycerol phosphate oxidase lost. In the first step of the reaction, 50 μl of reaction buffer was added to 50 μl of the supernatant from the lipolysis assay. A standard curve of 0-12,000 pmol was generated with standard glycerol diluted in permeate buffer. The plate was sealed and incubated at 37 ° C. for 30 minutes. The reaction resulted in the conversion of glycerol to dihydroxyacetone phosphate and hydrogen peroxide.

In the second stage of the reaction, the released hydrogen peroxide is measured using Quanta Blu fluorescence material (Pierce, Rockford, Ill.) And HRP, and the Quanta Blue wicking solution Was added to the sample and reference and 50 μl of 250 units / HRP ml was added. The Quanta Blue Wicking Solution contains 9 parts of Quanta Blue Matter Solution (Pierce, Quanta Blu Fluororgenic Peroxidase Substrate Kit, cat. # 15169) and pH 7.0 Kinase Buffer (2 mM MgCl ₂ , 100 1 part of mM triethanolamine, 2 mg / ml fatty acid-free BSA) was prepared by mixing. The plate was sealed and incubated at 37 ° C. for 30 minutes. The reaction was stopped by adding 50 [mu] l of Quanta Blu Stop solution (Pierce, Quanta Blue Fluorogenic Peroxide Date Kit, Cat. # 15169) per well. Relative fluorescence units (RFU's) of the product were measured at excitation and emission of 320 nm and 405 nm, respectively, and corrected for background fluorescence. Final data are presented as percent inhibition of lipid degradation. In the calculation, cAMP stimulation of lipolysis was considered 100% stimulus and vice versa 0% stimulus. The final data is expressed as percent inhibition calculated as follows.

RFU levels were corrected. In other words, detract from the background.

Example 10 Blood Free Fatty Acid Levels in Normal Mice Fasting Overnight

Dry male C57 / B16J mice were purchased from Jackson Laboratories (Bar Harbor, Maine, USA). Animals were kept in accordance with NIH guidelines and water and food were freely accessible from time to time. Test compounds of the present invention were suspended in 0.25% methylcellulose (weight / volume in deionized water) and stored at −20 ° C. in single use aliquots. Animals were administered once daily for two weeks via the intraperitoneal route of administration. After the last dose, the animals were fasted overnight. The next morning, 250 μl blood sample was placed through a replay orbital cavity into a tube containing EDTA saline and centrifuged at 1,200 × g for 15 minutes. The NEFA C diagnostic kit (WAKO Chemicals USA, Richmond, VA, USA) is used to analyze non-esterified free fatty acid levels (NEFA) in plasma fractions, and a BM Hitachi 911 clinical chemistry analyzer. Glycerol levels were analyzed with an improved triglyceride GPO-Trinder diagnostic kit (Sigma) for use. P-values from the Student's t-test were calculated using a Prism software data analysis program (Graphpad, Sorrento, Canada). The data is shown in Table 4 below. As can be seen from the table, animals using 2α-benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-olide at a dose of 50 mg / kg Treatment resulted in a 27% reduction in fasting NEFA plasma levels.

group NEFA, mM Glycerol, mg / dl NEFA Reduction% Vehicle 1.108 ± 0.193 36 0 2α-benzoyloxy-5,7,8αH-udesma-4 (15), 11 (13) -diene-8,12-oxide (50 mg / kg) 0.808 ± 0.080 ^a 28 ^b 27 A = p <0.0055 compared to vehicle b = p <0.05 compared to vehicle

Example 11 Effect on Fasting KK / Ay Mice

Male KK / Ay mice were purchased from Clea Japan USA (Fennington, NJ). Four-month-old animals were intraperitoneally administered 2α-benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide daily for two weeks. As described in Example 10, fasted plasma NEFA levels were obtained. Cholesterol and triglycerides were also measured using the BM Reagent Analysis Kit on the BM Hitachi 911 Clinical Chemistry Analyzer. The data show that NEFA, triglycerides, cholesterol and glycerol are significantly reduced in mice taking 2α-benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-olide. It was shown. Plasma glucose levels were measured using the Boehringer Mannheim Reagent Assay Kit on a Boehringer Mannheim Hitachi 911 Clinical Chemistry Analyzer. Control mice showed fasting plasma glucose levels of 184 ± 24 mg / dL, whereas animals receiving test compounds showed plasma glucose levels of 90 ± 29 mg / dL (p <0.01).

Example 12 Effect on Fasting ob / ob Mice

Male C57 / B16J ob / ob mice were purchased from Jackson Levatoritoris, Bar Harbor, Maine, USA. The 8-9 week old animals were administered compound daily via the intraperitoneal route for a two week period at the indicated dose. Test compounds of the present invention were prepared as previously described in Example 10. At 2 or 4 weeks of treatment, animals were fasted overnight and plasma NEFA levels were measured as described in Example 11. Data are presented as reduced percentages compared to animals that only received vehicle only. Test compounds led to a decrease in NEFA levels in treated animals. Glucose levels were measured using a Johnson & Johnson One-Touch Glucometer from 20 μl blood collected and sampled through a reparative orbital cavity with heparin-added microcapillaries. Fasting glucose levels for the vehicle group ranged from 250 to 350 mg / dL compared to 100 to 120 mg / dL of normal mice without diabetes. Data is expressed as normalized percentage using the following operation.

a) range = vehicle control value-normal value

b)% normalization = (vehicle control value-compound treatment value) / range

Example 13 Effect on Blood Free Fatty Acid Levels on Fasted ZDF Rats

Similar studies were performed on male ZDF rat models with type II diabetes purchased from Genetic Models, Inc., Indianapolis, Indiana, USA. The animals were intraperitoneally administered with the compound daily as described above. Plasma NEFA levels were obtained in the fasted state 6 weeks after treatment. As can be seen in the KK / Ay mouse model described in Example 11, a similar effect of lowering fasting NEFA levels occurred in ZDF rats.

Example 14 Effect on Insulin Resistance in KK / Ay Mice

KK / Ay mice were administered compounds as described in the examples above. Two weeks after treatment, overnight fasted animals are subjected to an oral glucose tolerance test (OGTT) in a fasted state by administering a 2 gm / kg glucose solution prepared with deionized water and measuring venous glucose from the tail tip at regularly scheduled times. . In addition, data obtained from young C57 / B16J mice without diabetes were included as comparative examples. The calculation of the area under the curve (AUC) shown in Table 2 was performed using a right-angle sum procedure incorporated into the Prism Data software analysis program (GraphPad, Sorrento, Canada). % Normalization of AUC was derived using the following operation.

a) range = vehicle control value-normal value

b)% = normalization = (vehicle control value-compound therapeutic value) / range

Mouse treatment with test compounds resulted in a significant normalization of glucose disposition to the application.

Example 15 Effect on Hyperglycemia Insulin Resistance in ob / ob Mice

Female ob / ob mice were administered the compound as described below. Five to six weeks after dosing, OGTT was performed as already described in Example 14. Initial fasting glucose levels were obtained from mice without diabetes. In the absence of prevailing fasting hyperglycemia defined as> 140 mg / D1 levels in the American Diabetes Association, which is clinically referred to as Syndrome X, the compound of the present invention is capable of disposing glucose in this condition. Significant normalization of.

All publications and references mentioned herein, including but not limited to patents and patent applications, are fully incorporated by reference in their entirety as if each individual publication or reference were specifically and individually referred to herein as being incorporated by reference in its entirety. Hereby incorporated by reference. All patent applications to which this application claims priority are also incorporated herein by reference in their entirety in the manner described above for publications and references.

While the invention has been described with emphasis on preferred embodiments, it will be apparent to those skilled in the art that modifications in the preferred apparatus and methods may be used and that the invention may be practiced otherwise than as specifically described herein. Accordingly, the present invention includes all modifications included within the spirit and scope of the invention as defined by the following claims.

Claims (17)

Lowering the blood lipid level or blood glucose of a mammal comprising administering to the mammal an effective amount of a lipid lowering or glucose lowering of the at least one compound of the AB ring structure of Formula I, or a pharmaceutically acceptable salt thereof How to let. <Formula I> Where The dotted line between carbon 5 and 6, the dotted line between carbon 4 and R ² and the dotted line between carbon 11 and R ³ independently represent each double bond from each numbered carbon to the linked carbon. , R ¹ is hydrogen, hydroxy, lower alkanoyloxy or aroyloxy group, and aryl of aroyloxy is at least one lower alkyl, lower alkoxy, halo, trifluoromethyl, di (lower) alkylamino, hydroxy, May be substituted with a nitro or C ₁ -C ₃ alkylenedioxy group, X is hydroxy or is oxy linking carbon 8 and carbon 12 with Y, Y is an hydroxy or amino group wherein lower alkyl, where lower alkyl may be bonded to form a 5- or 6-membered ring, may be mono- or di-substituted, R ² and R ³ are independently of each other a lower alkyl or alkenyl group, and lower alkyl of R ³ may be substituted with carbon in which an amino group, which may be mono- or disubstituted with lower alkyl, is linked to C 11, and the lower alkyls are bonded to 5 Can form a circular or six-membered ring, Provided that when R ¹ is hydrogen, (a) X is hydroxy or (b) R ³ is substituted with amino. The method of claim 1 wherein the hydrogen substituents at positions 5, 7 and 8 are in the α position relative to the A-B ring structure. The method of claim 1, wherein R ¹ is a lower alkanoyloxy group. The method of claim 3, wherein the alkanoyloxy group is selected from the group consisting of acetoxy, propanoyloxy, isobutyryloxy and cyclopropanoyloxy. The compound of claim 3, wherein the compound is selected from 2α-acetoxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-propanoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-cyclopropanoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-acetoxy-4,5,7,8,11αH-Eudesman-8,12-oxide; 2α-cyclopropanoyloxy-5,7,8,11αH-udesme-4 (15) -ene-8,12-oxide. The method of claim 1, wherein R ¹ is an aroyloxy group. The method of claim 6, wherein the aroyloxy group is selected from the group consisting of peroyloxy and benzoyloxy. 8. The compound of claim 7, wherein the compound is selected from 2α-benzoyloxy-5,7,8αH-Eudesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-perroyloxy-5,7,8αH-udesma-4 (15), 11 (13) -diene-8,12-oxide; 2α-benzoyloxy-4,5,7,8,11αH-Eudesman-8,12-oxide; And 2α-benzoyloxy-5,7,8,11αH-Eudesme-4 (15) -ene-8,12-oxide. The method of claim 1, wherein the compound is administered in a lipid lowering effective amount. The method of claim 1, wherein the compound is administered in a glucose lowering effective amount. The compound of claim 1 wherein (1) X is oxy, (2) R ¹ is lower alkanoyloxy or aroyloxy, provided that R ² is methylene linked to C4 by a double bond and R ³ is In the case of methylene linked to C11, R ¹ is not ethanoyl, and (3) R ² and R ³ are independently of each other a lower alkyl group or lower alkenyl group with the proposed unsaturation as linkage to carbon 4 or carbon 11 Way. The process of claim 1, wherein (1) X is oxy, (2) R ¹ is lower alkanoyloxy or aroyloxy, and (3) R ² and R ³ are independently of each other carbon 4 or carbon 11 A lower alkyl group or a lower alkenyl group having a limited unsaturation as the linkage. Compound of formula (I) or a pharmaceutically acceptable salt thereof. <Formula I> Where The dotted line between carbon 5 and 6, the dotted line between carbon 4 and R ² and the dotted line between carbon 11 and R ³ independently represent each double bond from each numbered carbon to the linked carbon. , R ¹ is hydrogen, hydroxy, lower alkanoyloxy or aroyloxy group, and aryl of aroyloxy is at least one lower alkyl, lower alkoxy, halo, trifluoromethyl, di (lower) alkylamino, hydroxy, May be substituted with a nitro or C ₁ -C ₃ alkylenedioxy group, X is hydroxy or is oxy linking carbon 8 and carbon 12 with Y, Y is an hydroxy or amino group wherein lower alkyl, where lower alkyl may be bonded to form a 5- or 6-membered ring, may be mono- or di-substituted, R ² and R ³ are independently of each other a lower alkyl or alkenyl group, and lower alkyl of R ³ may be substituted with carbon in which an amino group, which may be mono- or disubstituted with lower alkyl, is linked to C 11, and the lower alkyls are bonded to 5 Can form a circular or six-membered ring, Provided that when (1) X is hydroxy, R ¹ is not hydrogen; (2) when R ¹ is hydroxy, (a) X is hydroxy or (b) R ³ is substituted with amino, (3) When R ² is methylene linked to C4 by a double bond and R ³ is methylene linked to C11 by a double bond and X is oxy, R ¹ is not ethanoyloxy. The compound of claim 13, wherein X is hydroxy. 15. The compound of claim 14, wherein R ¹ is a lower alkanoyloxy or aroyloxy group. The compound of claim 13, wherein R ¹ is a lower alkanoyloxy group. The compound of claim 13, wherein R ¹ is an aroyloxy group.