IE46599B1 - Lipogenesis inhibitors - Google Patents

Abstract

(19) AU (21) 34701/78 (22) 3.4.78 (23) 3.4.78 (24) 4.4.77 (31) 784097 (32) 4.4.77 (33) US (43) 11.10.79 (51)2 A61K 31/335 A61K 31/40 A61K 31/445 C07D 303/02 C07D 405/06 (54) LIPOGENESIS INHIBITORS (71) SHELL INTERNATIONAL RESEARCH MAATSCHAPPIJ B.V. (72) DURHAM H.G.(74) SF (57) Method of inhibiting lipogenesis in warm-blooded animals is also COMPLETE SPECIFICATION(ORIGINAL) FOR OFFICE USE:-34701/78 [AU3470178A]

Description

This .invention relates to a composition useful as a lipogenenis inhibitor jn warm-blooded animal».

The invention provides a composition for inhibiting lipo^enesis in warm-blooded animals comprising a pharmaceut5 ically-acceptanle carrier and, in a pharmaceutically-acceptable state of purity, a 3-substituted-2-(aminocarbcnyl)-cxiranecarboxylic acid ester of the general famula: wherein R is alkyl, cycloalkyl, alkenyl or alkynyl, or is phenyl, methylphenyl, chlorophenyl, phenalkyl or cyclo10 alkylalkyl; R^ is alkyl, alkenyl, alkynyl or cycloalkyl; 2 . . and R and R each individually is hydrogen, one of the moieties represented by r\ or together represent a tetramethylene or pentamethylene chain forming a hetero ring with the indicated nitrogen atom.

In these compounds, each alkyl, alkenyl and alkynyl moiety can be of dr aight-chain or branched-chain configuration, and of up to twenty carbon atoms. Each cycloaikyl moiety can contain from three to six carbon atoms, while the alkyl portion of each cycloalkylalkyl moiety PO and each phenalkyl moiety can contain from one to six -3carbon atoms, preferably with from one or two carbon atoms joining the cycloalkyl or phenyl moiety to the oxirane ring.

Preferred of these compounds, because of their 5 activity as lipogenesis inhibitors, are those wherein R is phenyl or alkyl of from four to twelve carbon atoms, Ξ . 3 R and R each is hydrogen and R is alkyl of from one to four carbon atoms.

Compounds employed in the composition according to the invention can exist as either of two geometrical (cis-trans) isomers, depending upon the spatial relationship of the moieties upon the oxirane ring. Further, chirality exists in the compounds due to the asymmetric configurations at the 2- and 3-positions of the oxirane ring. As a result, four optical isomers exist, one pair for each of the two geometrical isomers. Both the cis and trans isomer products, as prepared, have been found to inhibit lipogenesis. In this specification, for the sake of simplicity, these compounds will, be referred to generally as 3-substituted-2-(aminocarbonyl)ox:iranocarboxylic acid esters this terminology including each of the isomers, as well as mixtures thereof. Under the circumstances, the invention contemplates each of the individual isomers, as well as mixtures thereof.

For illustration, preparation of typical compounds defined by formula I are described in tne examples 46S99 -4included hereinafter. Other typical, illustrative individual species of this genus are those wherein the symbols define the moieties: x . 12 R-' is ethyl, R and R each is hydrogen, R is dodecyl; x 12 R is ethyl, R and R each is hydrogen, R is butyl; x 1 2 R·, is ethyl, R is hydrogen, R is isopropyl, R is 2,6-dichlorophenyl; x 12 R is 2-propenyl; R = R = H; R χ 4-chlorophenyl; R-5 is 2-propynyl; R1 = R2 = H; R = phenyl; R^ is cyclohexyl; R1 = R2 = H; R = 4-methylphenyl; R^ is ethyl, R1 = H; R2 = 2-propenyl; R = phenyl; R^ is ethyl; R1 = H; R2 = 2-propynyl; R = 4-chlorophenyl; R-5 is ethyl; R1 = H; R2 = cyclohexyl; R = octyl; R^ is butyl; R^ + R2 = -(CH2)-^; R = hexyl; R^ is propyl; R1 = R2 = H; R = 2-pentenyl; R-5 is ethyl; R1 = R2 = H; R = benzyl; R^ is ethyl; R^ = R2 = H; R = 4-pentynyl; r3 is ethyl; R1 = R2 = H; R = cyclohexylmethyl.

The lipogenesis inhibitors employed in the composition I according to this invention are a known class of compounds, those wherein R is alkyl being disclosed by M.Igaroshi and H. Midorikawa, J. Org. Chem., 28, 5088-3092 (1963) (Reference J), while those wherein R is phenyl are disclosed by A. Robert and A. Foucaud, Bull. Soc. Chim.

France, 7_, 2531 (1961) (Reference II). Compounds contemplated in the invention not specifically disclosed in those publication:: cnn h- pn.’parfd by tin- method:: disclosed therein.

The 3-substituted-2-(aminocarbonyl)oxiranecarboxylie acid esters of formula I can be used to control lipogenesis in warm-blooded animals such as, for example, pets, animals in a zoo, livestock, fur-bearing animals, including, but not limited to dogs, cats, mink, sheep, goats, swine, cattle, horses, mules, donkeys and poultry. The effect is obtained by administering an effective amount of one or a mixture of two or more of the compounds of formula I orally or parenterally to the animal. They may be administered as such, or as an active ingredient of a conventional pharmaceutical formulation. Accordingly the present invention includes a method of inhibiting ligogenesis in non-human warm-blooded animals which comprises administering to such an animal an effective amount of a compound of formula I or a composition containing it.

The compounds or compositions containing them may be administered orally by any convenient means. Thus, they may be orally administered as a drench, by intubation, in the animal's food and water, in a food supplement or in a formulation expressly designed for administration of .the drug. Suitable formulations include solutions, suspensions, dispersions, emulsions, tablets, boluses, powders, granules, capsules, syrups and elixirs. For parenteral administration, they may -(ibe in the form of a solution, suspension, dispersion or emulsion. They can be administered in the form of an implant or other controlled sustained release formulation. Inert pharmaceutically-acceptable carriers, such as one or more of water, edible oil, gelatin, lactose, starch, magnesium stearate, talc or vegetable gum can be used.

The dosage of the compound of formula I needed to inhibit lipogenesis will depend upon the particular compound used, and the particular animal being treated. However, in general, satisfactory results are obtained when the compounds are administered in a dosage of from about 1 to about 500 milligrams per kilogram of the animal's body weight. It can be administered in a single dose or in a series of doses in the same day, or over a period of days. For any particular animal, I a specific dosage regimen should be adjusted according to the individual need, the particular compound used as the inhibitor, and the professional judgement of the person administering or supervising the administration of the inhibitor.

The invention is further illustrated by the following Examples. In the preparative Examples the identities of the product, and of the precursor(s) involved were confirmed by appropriate chemical and spectral analyses. 46S99 EXAMPLE 1 - Ethyl 2-( aminocarbonyl)-3_htxyloxirane carboxylate (trans)(1) JL was prepared as a white crystalline solid, m.p.: 86°C by the procedure of Reference jl, using sodium tungstate dihydrate as catalyst.

EXAMPLE 2 - Ethyl ,2-(aminocarbonyl)-3~hexyloxirane carboxylate (cis)(2) A mixture of 31 g of diethyl heptylidenepropanedicarboxylate (B. Wojcik and H. Adkins, J. Am. Chem.

Soc., 56, 2424 (1954), prepared by the method of A.C. Cope and K.E. Hoyle, J. Am. Chem. Soc., 63, 733 (1941)), 5 g of potassium bicarbonate and 17 g of 30% hydrogen peroxide in 200 ml of methanol was stirred for two hours at 40°C, then was allowed to stand at 20°C for l6 hours. The mixture then was concentrated to 50 ml; 100 ml of water was added and the solution was extracted with two 50 ml volumes of methylene chloride. The solvent was evaporated to give a colourless liquid, which was vacuum distilled to give diethyl 3"hexyloxiranedicarboxylate (2A),b.p.: 117-126°C at 0.1 Torr.

A solution of 8.0 g of 2A, in 10 ml of ethanol was saturated with ammonia (gas) and the mixture was allowed to stand for 2 days at room temperature. The precipitate that formed was collected and recrystailized from pentane to give 2, as white crystals, m.p.: 58-6O°C. -8£6 59 9 EXAMPLE 3 - Ethyl 2-(aminocarbonyl)-3-phenyloxirane carboxylate(5) was prepared as a white crystalline solid, m.p.: 15O-151°C by the procedure of Reference II.

EXAMPLE 4 - Ethyl 2-(aminocarbonyl)-3_methyloxirane carboxylate(4) g of ethyl 2-cyano-2-butenoate (P.D. Popp et al., J. Org. Chem., 26, 2738-40 (1961)) and 2 g of sodium tungstate dihydrate were dissolved in 100 ml of ethanol. ml of 3052 hydrogen peroxide was added to the solution dropwise at 50°C. A vigorous exothermic reaction occurred and the mixture was cooled to maintain it below 70°C, When the reaction was complete, the solvent was evaporated and the residue, an oil, was extracted with hot cyclohexane. The residue was distilled to give ethyl 2-cyano-3~ methyloxiranecarboxylate (4A), as a fraction boiling at 67-68°C at 0.1 Torr. g of 4A, 2 g of sodium tungstate dihydrate and 2 g of trisodium phosphate were mixed with 30 ml of ethanol. 50 ml of 30$ hydrogen peroxide was added over a 30 minute period to the stirred mixture at 55°C and the mixture was stirred for 90 minutes while being warmed to 60°C. Then 35 ml of 30$ hydrogen peroxide was added at 60°C, sufficient ethanol to make the mixture homogeneous was added and the mixture was stirred for 5 hours at 60°C. The solvent then was evaporated and the aqueous residue 46509 -9was extracted with methylene chloride. The solvent was evaporated from the separated extract and the residue was triturated with carbon tetrachloride to give a solid product, which upon recrystallization from chloroform gave 4, as a white solid, m.p.: 139-140°C.

EXAMPLE 5 - Ethyl 2-((methylamino)carbonyl)-3-ptenyl oxiranecarboxylate(5) g of benzalmalonic ester (E.H. Kroeker, et al., j. Am. Chem. Soc., 56., 1171-3 (1934)) and 10 ml of 10# aqueous sodium bicarbonate were mixed with 100 ml of ethanol. 42 ml of 30% hydrogen peroxide was added dropwise over a 5-hour period at 70°C. Then 30 ml of 30% hydrogen peroxide was added over a one-hour period at 60°C. 100 ml of ethanol was added and the mixture stirred for 8 hours at 60°C. The solvent was evaporated and the residue extracted‘with methylene chloride. The solvent was evaporated from the separated extract and the residue was distilled to give ethyl 2-(ethoxycarbonyl)-3~phenyloxiranecarboxylate (5A), as a fraction boiling at 173-lSO°C at 0.05 Torr. g of 5A and 4 g of 40% aqueous methylamine were mixed and the mixture was warmed to 30°C. Then 2 g of ethanol was added and the mixture was allowed to stand for 105 minutes. The solvent was evaporated. The residue was poured into water and the oily layer which formed was separated and treated with ether to leave 5, as a whitf solid, m.p.: 1O2-1O3°C. -10EXAMPLE 6 - Tests to demonstrate lipogeneuis inhibition ι The effectiveness of the 3-substituted-2-(aminocarbonyl )oxiranecarboxylic acid esters was ascertained by Immersing samples of animal liver or adipose tissue in a liquid medium containing radioactive glucose and the test chemipal, for a period of time, then isolating the lipid from the treated tissue and determining the uptake of the radioactive carbon by means of scintillation counting techniques. These tests were conducted in swine adipose tissue, because in swine the primary site of lipogenesis appears to be adipose tissue.

Described in more detail, the tests were conducted according to the following general procedure. 150 milligrams of slices of swine adipose tissue 15 were incubated at 37°C for 2 hours with shaking in 3 millilitres of Krebs-Ringer bicarbonate solution containing one-half the normal calcium ion concentratbn, micromoles of glucose, 0.5 micro-Curie of glucose-U C, and 300 micro-units of insulin, and 5$ dimethyl sulphoxide 2θ (DMSO). The test compounds were added as suspensions or solutions in DMSO and were present at a concentration of 100 micrograms per millilitre of the incubation mixture.

The incubation was terminated by addition of 0.25 ,’5 millilitre of 1 N sulphuric acid. The resulting mixture was extracted with a total of 25 millilitres of chloro-1146S99 form:methanol (2:1, v/v). The extracts were washed according to Folch et al. (J. Biol. Chem., 226, 497-509, (1957)) air-dried, and counted in a liquid scintillation counter with 15 millilitres of counting fluid (two parts toluene containing 0.4% w/v New England Nuclear Omniflutx: 1 part Trite» Χ-lOO). (Triton is a Registered Trade Mark) The tests were conducted in triplicate and were accompanied by control tests in which all-ingredients, proportions and conditions were the same except that no test compound was included. From the data obtained were calculated the percent inhibition of lipid synthesis by the test compound in each case.

The data obtained from the tests are set out in Table I, as the percent inhibition of lipogenesis compared to the results obtained in the control tests wherein only the test compound was omitted. (>impound No.

TABLE I Percent inhibition

Claims (6)

1. A composition for inhibiting lipogenesis in warmblooded animals comprising a pharmaceutically-acceptable carrier and, in a pharmaceutically acceptable state of purity, a 5 3-substituted-

2. -(aminocarbonyl)-oxirane carboxylic acid ester of the general farnula: wherein R is alkyl, cycloalkyl, alkenyl or alkynyl, or is phenyl, methylphenyl, ehlorophenyl, phenalkyl or lo cycloalkylalkyl; R-5 is alkyl, alkenyl, alkynyl or cyclo1 2 alkyl; and R and R is each individually hydrogen, one of the moieties represented by r\ or together represent a tetramethylene or pentamethylene chain forming a hetero ring with the indicated nitrogen atom. /5 2. A composition according to Claim 1, wherein in the compound of formula I each alkyl, alkenyl or alkynyl moiety contains up to 20 carbon atoms, each cycloalkyl moiety contains

3. To 6 carbon atoms, and the alkyl portion of each cycloalkylalkyl moiety and each phenalkyl moiety zo contains from 1 to 6 carbon atoms. -133. A composition according to Claim 1 or 2, wherein, in the compound of formula I, R is phenyl or alkyl 1 2 containing four to twelve carbon atoms, R and R are •5 both hydrogen and R is alkyl containing one to four 5 carbon atoms.

4. A composition according to Claim 1 including a compound of formula I specifically named herein.

5. A composition according to claim 1 substantially as hereinbefore described and with reference to 10 Example 6.

6. . A methed of inhibiting lipogenesis in non-human warm-blooded animals which comprises administering to such an animal an effective amount of a compound of formula I or a composition according to any one of Claims 1 to 5·