DE2704744C2 - Fatty acid amides of α-methyl-m- (trifluoromethyl) -phenethylamine, processes for their preparation and pharmaceuticals - Google Patents

Description

CF ₃ ^H

wherein R is saturated or unsaturated alkyl radicals having 12 to 22 carbon atoms

2. N- [1-methyl-2- (3-trifluoromethylphenyl) ethyl] octadeca-9,12-dienamide-cis-cis isomer.

3. N- [1-methyl-2- (3-trifluoromethylphenyl) ethylioctadec-9-enamide cis isomer.
4. N-il-methyl-l - ^ - trifluoro-methylphenyOäthyljoctadec ^ -enamid-trans-isomer.

5. N-tl-Methyl ^ -O-trifluoromethylphenyOäthyljoctadecanarnid.

6. A mixture of fatty acid amides according to claim 1, wherein the predominant fatty acid amide is I noleic acid amide.

7. Process for the preparation of the fatty acid amides according to Claim 1 to 6, characterized in that one in a known manner to a solution of a-methyl-m- (trifluoromethyl) -phenäthy3aimin in dry Phosphorus trichloride is added to pyridine while stirring and cooling with an ice bath and then the solution with a fatty acid having 12 to 22 carbon atoms in the alkyl group or a mixture of such fatty acids added and heated.

8. Medicaments consisting of fatty acid amides according to one of claims 1 to 6 and customary carriers.

The invention relates to new fatty acid amides of ff-methyl-m- (trifluoromethyl) phenethylamine formula

CH

<f S-CH ₂ -CH-NH ₂

CF ₃

which is given the trivial name Norfenfluramine in the literature (M. Negwer, Oerganisch-Chemische Medicines, Vol. 1, p. 182, 1971) and is also referred to as follows.

Atherosclerosis, which is a form of atherosclerosis that is characterized by clogging of the blood vessels partly due to the deposition of excess lipids from the bloodstream, mainly cholesterol and triglycerides, which circulate through the body associated with proteins called lipoproteins, with cholesterol appearing to be the bigger threat. A decrease in the concentration of lipids in the blood appears a desirable agent for combating atherosclerosis and related heart diseases to be. Lipid levels in the blood can be determined by any of the following methods or through a combination of different methods depending on the primary type of hyperlipemia limiting lipid intake, through food selection, through voluntary dietary requirements Restriction of food in general, through the administration of anorectic Medicines that restrict food intake in general, or through the use of medicines, known as anti-hyperlipidemic agents are decreased. When using a antihyperlipidemic agents can generally be said that no single drug is completely effective with regard to the lowering of all classes of serum lipids and that no drug is uniform is effective in lowering cholesterol and triglycerides. Accordingly, there is a Need for more effective drugs to fight atherosclerosis.

Anti-hyperlipidemic agents are believed to act in one way or another in terms of degradation of serum cholesterol as follows:

(1) Inhibition of the synthesis of cholesterol in the liver and / or in the ilium (ileum),
(2) inhibiting the absorption of dietary cholesterol,

(3) Promoting excretion through increased catabolism of cholesterol from circulating lipids and from extrahepatic tissue.

The association of atherosclerosis with high serum cholesterol levels has made treatment for this disease directed to agents that lower serum cholesterol. Cholestyramine (Polystyrenetrimediylbenzylam-

monium chloride) and clofibrate (ethyl p-chlorophenoxyisobutyrate) act selectively with regard to the reduction of cholesterol, primarily through impairment or disturbance of its alimentary absorption or its syn-

Other known compounds, the long chain fatty acid amides of certain α-methylbenzylamines and -anilines, which are ascribed cholesterol-lowering activity, are disclosed in U.S. Patents 3,621,043 and 37 28 459, the ar-methylbenzyllinoleamide being of particular interest

Obesity (obesity) is often closely related to atherosclerosis and can be characterized by the same basic anomaly in the chemistry of individuals or mammals.

The invention is based on the unexpected finding that certain fatty acid amides of norfenfluramine (e-methyl-m- (trifluoromethyl) phenethylamine of the formula

CH ₃

J ~%, I.

<f V-CH ₂ -CH-NH ₂

in terms of lowering cholesterol not only by reducing its absorption, but also also affect its synthesis and that the nutritional efficacy as well as the appetite are reduced, to bring about a reduction in weight gain.

Thus, the compounds according to the invention have multiple functions in terms of controlling lipid levels in the blood tjm, as follows:

(1) appetite suppressant, which decreases cholesterol absorption,

(2) reducing cholesterol absorption and

(3) decrease in cholesterol synthesis in the liver.

One of these functions, (3) reducing cholesterol synthesis in the liver, is not characteristic for the α-methylbenzylamides of the prior art, and the function (2), (Ve reduction of liver and Serum cholesterol is more pronounced with compounds according to the invention in the hypercholesteremic Diet. In addition, the compounds differ from the known amide compounds in that the Amides according to the invention reduce weight gain in two ways, i. H. (1) through Decrease in appetite and (2) decrease in food efficiency. While in the US-PS 36 21 043 stated that the known compounds do not affect appetite, was in the "Journal of Atherosclerosis Kdsearch", Vol. 9, pages 65 to 71 (1969) reports that a known compound (ff-Methylbenzy'linoleumid) suppresses the increase in the body weight of rats, which is due to decreased Food intake appears to be the cause. As explained below, it can be seen that in addition to suppressing appetite, the compounds according to the invention increase food efficiency decrease, a function that is not apparent in the known connections tested is.

Another distinction from the amides of the prior art is the greater degradation with regard to low density lipid carriers in blood serum. It became a decrease with a decrease the lipid carrier of high density with increasing concentration when using compounds according to FIG Invention observed.

The compounds according to the invention are therefore more suitable for controlling lipid levels in blood serum and have a beneficial effect in combating obesity, which the amide compounds of the State of the art do not have.

The invention relates to new amides of norfenfluramine, processes for their preparation and compositions for lowering elevated serum cholesterol levels in animals of the mammalian species with a hypercholesteremic diet were taken care of. The compounds act in multiple ways in terms of Reduction of serum and liver cholesterol and are ideally suited for combating atherosclerosis of those suffering from it. In addition, dit compounds retain some anorectic activity Parent amine, d. H. des Norfenfiuramine without any of the undesirable side effects the Accompany administration of norfenfluramine. In the same way, the amides of norfenfluramine affect the use of the food given in order to reduce the expected weight gain. Thus, in addition to lowering cholesterol, the effects of anorexia come together (Loss of appetite) and decreased food utilization to reduce weight gain without any noticeable effect on the well-being of people or other living beings and only weakly affect the appetite. You are therefore in terms of Effectively combating obesity in persons and living beings afflicted with hypercholesteremia are. Change studies show that the concentration of norfenfluramine units of the fatty acid amides in the Blood is low considering the amount of drug given.

The fatty acid amides according to the invention have the general formula

CH ₂ -CH-NCR

(D

CF ₃ ^H

wherein R is saturated or unsaturated alkyl radicals having 12 to 22 carbon atoms.

The compounds according to the invention act as cholesterol-lowering agents with beneficial influence in fighting as obesity. Especially the long-chain fatty acid amides of norfenfluramine have a dual effect of lowering elevated cholesterol levels in humans or mammals on a high cholesterol diet by impairing or hindering the absorption of cholesterol and the synthesis of cholesterol in the liver and a complementary activity in reducing the Weight gain, with the decrease in weight gain partly due to decreased appetite and, in part, to decreased food efficiency from ingested foods is due. The cholesterol lowering agents according to the invention are ideally suited for Fight the diseases atherosclerosis and hypercholesteremia without harmful side effects and to combat obesity in persons or mammals afflicted with these diseases are.

Due to the easy accessibility, the good tolerance and the excellent combination of effectiveness the N- [l-MethyI-2- (3-trifluoromethylphenyl) ethyl] octadeca-9,12-dienamide-cis-cis isomer, the N-fl-Methyl ^ -ß-triflüormethylphenyOäthylloctadec ^ -enamid-cis-isomer, the N- [l-MethyI-2- (3-trifluormethylphenyOäthylJoctadec-Q-enamid-trans-isomer, the N-tl-methyl-1-O-trifluoromethylphenyOä'thylJoctadecanamid and mixtures of fatty acid amides according to the invention, especially those in which linoleic acid amide the predominant fatty acid amide is preferred.

The preparation of the amides of norfenfluramine according to the invention can be carried out by one of the customary processes for the preparation of acid amides, such as
20th

(1) Reaction of norfenfiuramine in dry pyridine with phosphorus trihalide to form a phosphazo compound to form, with the following?:? Reaction with the fatty acid or a mixture of fatty acids, what is the preferred method.

(2) Reaction of norfenfluramine with fatty acid halide or a mixture of fatty acid halides, (3) Conversion of fatty acids or a mixture of fatty acids, norfenfluramines and suitable disubstituted ones Carbodiimide,

(4) Implementation of fatty acids or a mixture of fatty acids with norfenfiuramine at 100 to 300 ⁰ C and

(5) catalytic dehydration of fatty acid or mixtures of fatty acids and norfenfiuramine via a dehydrating agent such as sulfuric acid, p-phenolsulfonic acid or the like, or a cation exchange resin.

The acids used in the preparation of the amides according to the invention can be of any origin

However, the acids usually come from some natural fats and oils, in particular Vegetable oils, such as tall oil, linseed oil, hemp seed oil, safflower oil, soybean oil, sunflower oil, rice

shell oil, corn oil, tree seed oil, olive oil, peanut oil, palm oil, coconut oil, and animal oils found in the most animal fats and fish oils can be found.

The invention is explained in more detail below with reference to examples, which the production and the physical Show constants of the preferred amides.

example 1

(Linoleamide from Norfenfluramine)
Nn-Methyl ^ -Q-trifluoromethylphenyOäthylloctadeca - ^^ - dienamide (cis-cis)

A mixture of 61.56 g (0.18 mol) of norfenfluramine hydrochloride in chloroform was treated with a 10% strength extracted aqueous solution of sodium hydroxide. The chloroform layer was dried with sodium sulfate, filtered and concentrated under vacuum. The residue was dissolved in one liter of dry pyridine and it 15.0 g (0.11 mol) of phosphorus trichloride were added dropwise while cooling with an ice bath and while stirring admitted. After stirring for one hour at room temperature, 50.0 g of pure linoleic acid (0.18 mol) were added. This solution was refluxed for 2 hours. After cooling the solution was taking Vacuum concentrated. The residue was partitioned between chloroform and a 10% solution of sodium hydroxide divided up. The chloroform layer was dried with sodium sulfate, filtered, concentrated in vacuo and distilled. Bp 225-230 ° / 0.1 mm Hg; the yield was 44.0 3 (0.09 mol); 50% of theory.

Analysis for C ₂₈ H ₄₂ F ₃ NO:
Calculated: C 72.23 H 9.09 N 3.01
Found: C 72.37 H 9.08 N 2.97
60

Example 2

(Oleamide from Norfenfluramine) N-tl-Methyl ^ -O-trifluoromethylphenyDäthylloctadec ^ -enamid-cis-isomer

A mixture of 61.56 g (0.18 mol) of norfenfluramine hydrochloride in chloroform was with a IO% sodium hydroxide solution extracted. The chloroform layer was dried with sodium sulfate, filtered and concentrated under vacuum. The residue was dissolved in 1 1 of dry pyridine, and 15.0 g Phosphorus trichloride (0.11 mol) was added with stirring and cooling with an ice bath. After an hour Stirring at room temperature, 50.0 g of pure oleic acid (0.18 mol) was added and the solution was left for 15 h boiled under reflux conditions. After cooling, the solution was concentrated in vacuo and the Residue was partitioned between chloroform and a 10% solution of sodium hydroxide. The chloroform layer was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was distilled. Bp 230-235 ° / 0.1 mm Hg; the yield was 42.0 g (0.09 mol); 50% of theory.

Analysis for C ₂₈ H ₄₄ F ₁ NO:
Calculated: C 71.9! H 9.48 N 3.0?
Found: C 71.90 H 9.43 N 2 ^ 98

Example 3
(Elaidic acid amide from norfenfluramine)

N-n-Methyl ^ -Q-trinuormethylphenyDäthylloctaaec ^ -enamid-trans-isomer

Following the procedure of Example 2 except that 50.0 g (0.18 moles) of elaidic acid were used of oleic acid were used, the above-mentioned compound was obtained in 15% yield. Bp 230-235 ° / 0.1 mm Hg.

Analysis for C ₂₈ H ₄₄ F ₃ NO:
Calculated: C71.91 H 9.48 N 3.00
C 71.83 H 9.40 N 2.99

Example 4
(Stearamide from Norfenfluramine)

N- [1 -Methyl ^ -O-trifluoromethylphenyOethylJoctadecanamid

Following the procedure of Example 2 with the modification that oleic acid is replaced by 50.0 g (0.18 mol) of stearic acid was replaced and that the concentrated final residue was recrystallized from ethyl acetate instead of distilled, the above-mentioned compound was obtained in a yield of 29%; F 74-76 ° C

Analysis for C ₂₈ H ₄ ^ F ₃ NO:
Calculated: C 71.61 H 9.87 N 2.98
Found: C 71.61 H 9.69 N 2.90

Example 5

(Linoleamide from Norfenfluramine from raw linoleic acid)

A mixture of 36.7 g (0.1 mol) of norfenfluramine hydrochloride in chloroform was treated with a 10% strength extracted aqueous solution of sodium hydroxide. The chloroform layer was dried with sodium sulfate, filtered and concentrated under vacuum. The residue was dissolved in a 1 1 dry pyridine, and it 8.0 g (0.06 mol) of phosphorus trichloride was added dropwise while cooling with ice water and stirring admitted. After stirring for one hour at room temperature, 14.0 g (0.05 mol) of linoleic acid, the

Wt .- "/ Linoleic acid 65.5 Oleic acid 19.0 Linolenic acid 10.5 Palmitic acid 3.5 Myristic acid 0.5 Stearic acid 0.5 various fatty acids 0.5

100.0

contained, admitted. This solution was refluxed for two hours. After cooling the solution was concentrated in vacuo. The residue was between chloroform and a 10% Sodium hydroxide solution divided. The chloroform layer was dried with sodium sulfate, filtered and concentrated under vacuum. The remainder was dissolved in benzene and there were 75 ml aliquots on one Magnesium silicate column collected, with benzene and then with a 10% solution of acetone and benzene eluted. The portion containing the product was concentrated under vacuum and distilled. Bp 220 ° / 0.05 mm Hg; the yield was 26% of theory.

Analysis:

Found: C 71.94 H 9.15 N 2.84

Analysis using high pressure liquid chromatography indicated that the product had a Mixture of fatty acid amides from Norfenfluramine was and that the predominant amide mainly made up Linoleamide (linoleic acid amide) consisted of Norfenfluramine.

The fatty acid amides according to the invention of the formula I can in particular humans and mammals which suffer from hypercholesteremia or atherosclerosis. The compounds can be in the form of Capsules, tablets, syrups, elixirs or admixtures with food can be administered orally. When the compounds are administered as capsules, tablets, elixirs, and syrups, they generally should with meals in daily amounts of 50 to 600 mg / kg, preferably 200 to 600 mg / kg. When the compounds are administered as admixtures in foods, the weight percentage will vary of the compound, based on the food, from 0.025 to 0.5% by weight, preferably from 0.1 to 0.2 wt%. In general, humans and mammals show a favorable propensity at these dosages to reduce weight gain and reduce obesity.

pharmacology
Triton-induced hyperlipidemia tests

Caged male Sprague-Dawley rats weighing 250-350 g were fed ad libitum with commercially available laboratory chow with free access to water. The animals were locked up overnight and given Triton-WR-1339 (manufactured by Ruger Chemical Company, Inc. Irvington, New Jersey) as described by Schurr et al. In "Lipids" 7,68 (1972), with the modification that the saphenous vein was taken and divided doses of 70 mg / kg were used. The rats were anesthetized with sodium pentobarbital and bled by cardiac puncture. The serum was stored at -20 ⁰ C, was analyzed until analyzed for total cholesterol and triglycerides. A comparison with clofibrate was also made. As shown in Table I, the norfenfluramine fatty acid amides decreased serum triglycerides and serum cholesterol.

Diet-induced hyperlipemia tests

General feeding experiments on rats were carried out to assess the effect of the present invention Amides on serum and liver lipids from animals on a basic diet and one with cholesterol supplemented basic diet were fed. Also, the effects of these drugs have been assessed Serum lipoproteins and their effect on body weight, gross feeding efficiency, liver weight, Appetite and energy utilization examined with regard to their metabolic size. Male Sprague-Dawley Rats weighing 150 to 175 g were placed in cages with raised wire floors in a room that was maintained at a temperature of 24 to 26 ° C, with a 12-hour day and a 12-hour Night rhythm accommodated and fed ad libitum with a semi-purified diet, the following composition exhibited:

Calcium 7.1 chlorine 3.95 copper 0.0059 iodine 0.00067 iron 0.0287 magnesium 0.528 manganese 0.0527 phosphorus 4.55 potassium 4.35 sodium 2.40 sulfur 0.73 zinc 0.135

Wt%

vitamin-free casein

(ilucose

hydrogenated coconut oil GBI Vitamin Fortification Mixture, Cat. No. 40 060 from the General Biochemical Company, Chargrin Falls, Ohio, USA modified salt mixture from Williams and Briggs, Cat. No. 170 911 from the General Biochemical Company

Cellulose 2

and watered. The modified salt mixture provided the following amounts of elements in g / kg.

20th 25th

The vitamin fortification mix consisted of the mixture in g / kg:

p-aminobenzoic acid 11.0132

Ascorbic acid, coated 97.5% pure 101.6604

Biotin 0.0441

Vitamin B ₁₂ , 0.12% in mannitol 2.9736

Calcium pantothenate 6.6079

Choline Dihydrogen Citrate 349.6916

Folic acid 0.1982

Innosite 11.0132

Menadione (Vitamin K) 4.9559

Niacin 9.9il9

Pyridoxine HCL 2.2026

Riboflavin 2.2026

Thiamine HCL 2.2026

dry vitamin A palmitate (500,000 units / g) 3.9648

dry vitamin D ₂ (500,000 units / g) 0.4405

dry vitamin E acetate (500,000 units / g) 24.2291

Remainder, thinning corn starch 466.6878

The diets were supplemented with 0.5% cholic acid, 1% cholesterol and drugs at the expense of glucose. The test period was 18 to 21 days, with body weight and food being measured at specific time intervals. The animals were locked up overnight and bled by cardiac puncture. The blood was coagulated with anti-ethylene diamine, and the plasma was separated by centrifugation and stored at -20 ^0C. The liver was excised, weighed and stored at -20 ⁰ C. All samples were held at this temperature until they were analyzed for total lipids, total cholesterol, triglycerides and phospholipids. Gross feed efficacy was determined along with the effect of metabolic size on food intake and the effect of drug on energy utilization.

60

Table I.

Effect of fatty acid amides of norfenfluramine on serum triglycerides and cholesterol in trilonindu / hyperlipidemic rats

Group connection

Dose Serum-% An Seruirt-

mg / kg triglycerides lowering cholesterol

p.o. mg / 100 ml tri- mg / 100 ml

+ SD glyceride ± SD

% Change Cholesler

none (control) clofibrate linoleic acid amide (ex. 1)

none 588.2 ± 338.6 0

124.5 ± 7.8 -79 268.7 ± 69.6 -54

none (control) none

Clofibrate

Linoleic Acid & Mixed Acid Amides (Ex. 5)

231.0 ± 59.9 173.0 ± 54.0 209.0 ± 16.4

123.5 ± 13.4

94 ,! ± 13.6

101.9 ± 20.4

-24.0 -9.5

-24 -18

none (control) clofibrate

Linoleic acid & mixed acidic amides (Ex. 5) the same.

none (control) clofibrate

Oleic acid amide (Ex. 2) from Norfenfluramine

Elaidic acid amide (Ex. 3) from Norfenfluramine

Stearic acid amide (Ex. 4) from Norfcnflurarr.inc

140

140

166.8 ± 82.0 0

132.7 ± 64.5 -20 67.3 ± 19.1 -60

(a) abnormal result

64.3 ± 33.9 -62

(a) 600.0 ± 214.7 +260

none 618.3 ± 302.2 0

411.7 ± 118.0 -34

166.3 ± 37.2 -73

260.0 ± 137.8 -58

160.0 ± 110.0 -74

Basic diet

140.0 ± 20.0

109.3 ± 24.4 -22

109.0 ± 10.0 -22

109.7 ± 16.6 -22

189.0 ± 13.2 +28

281.5 ± 40.3

213.0 ± 14.1 -24

173.3 ± 10.2 -39

210.0 ± 47.8 -25

214.3 ± 91.6 -24

The results of testing the mixed amides of Example 5 and the reference compounds in Rrs'en at a basic diet according to the procedure described above for diet-induced hyperlipemia are disclosed in US Pat Tables 2 and 3 given. Notably, the amides of the invention (Example 5) decreased food intake and the feed efficiency is considerable, d. H. lower weight gain per unit taken Food.

Basic diet with cholesterol

The results of testing the mixed amides of Example 5 and the reference compounds in rats in the case of a basic diet with additional cholesterol according to the procedure described above for diet-induced diets Hyperlipemia are shown in Tables 4 and 5. The mixed amides of Example 5 produced again a considerable decrease in feed efficiency, but also a decreased serum and liver cholesterol observed.

Data similarly obtained in Table 6 using the pure linoleamide from Norfenfiuramine of Example 1 also show the effect on serum and liver lipids.

In other analyzes, the increased doses of mixed amides of Example 5 produced decreases Levels of high density lipoproteins (HDL) and continued decreasing levels of lipoproteins lower Density (LDL), see Table 7.

Toxicity comparison

The motor activity of norfenfluramine and the amides of norfenfluramine according to the invention were determined by modification of the activity in female Aibinornäusen of the ICR-Starnmes the method of Cook et al in J. Pharmac. Exp. Ther. 113: lla (1955) compared. The results point to this suggest that the amides of norfenfluramine have only a weak effect on motor activity in one Dose from 100 to 300 mg / kg, i.v. p., compared to the strong effects of Norfenfluramine in

a dose of 6.5 mg / kg, i. p. was established. In mice, at doses of 300 mg / kg, L p. and 1000 mg / kg, p. o-, no neurotoxicity with the amides of Example 5, while at a dose of 40 mg / kg, i. p., with norfenfluramine tremors and at a dose of 100 mg / kg, i. p., death occurred

Table II

Effects on liver weight, body weight and food consumption in rats
(fed with semi-purified basic diet)

Treatment ¹ · ') Dose ² )
mg / kg /
Day number
Rats Liver weight,
Total g + SD
100 g each
Body weight Body weight,
Beginning g ± SD
end Close-
foresight
middle-
consumption,
G Gross-
futier-
effective
samk. ^b ) No 6th 5.7 ± 0.3 2.7 ± 0.3 158 ± 18 212 ± 22 309 0.175 Clofibrate 0.2% 209 6th 9.2 ± 0.4 4.4 ± 0.4 155 + 16 210 ± 21 306 0.180 Cholestyramine, 3% 3120 6th 5.8 ± 0.3 2.7 ± 0.3 151 ± 8 215 ± 13 316 0.203 Amides before Norfenflur
amide (ex. 5) 0.1% 96 4th 5.3 ± 0.8 2.8 ± 0.2 176 ± 21 190 ± 28 241 0.058 0.2% 204 3 7.0 ± 0.2 3.4 ± 0.7 171 ± 15 205 ± 22 304 0.112 0.5% 490 4th 5.9 ± 0.6 2.8 ± 0.1 185 ± 16 208 ± 24 276 0.083 ff-methylbenzyl-
lino'eamide 0.2% ^d ) 182 6th 6.7 ± 0.8 2.9 ± 0.3 158 ± 20 226 ± 30 331 0.210

^a ) = Based on "!. Medicines in the diet and average daily food consumption.

^h ) = gross weight increase in g, total food consumption in g.

^c ) = weight percent of additive in food.

^d ) = connection according to US-PS 36 21 043 and US-PS 37 28 459.

25th

30th

Table III

Effect on mean concentration of serum and liver lipids in rats
(fed with a basic diet cleansed by the skin)

Treatment * ¹ ) Lipids

Serum mg / 100 ml ± SD total cholesterol

Triglycerides Liver mg / g ± SD Total Cholesterol Triglycerides

40

no
Clofibrate, 0.2%

Cholestyramines

3%

Amides of
Norfeniluramine
(Ex. 5)

0.1%
0.2%
0.5%
σ-methylbenzyl

138.80 ± 22.57
143.70 ± 22.42
116.60 ± 23.16

153.30 ± 20.10
191.00 ± 49.80
175.50 ± 9.40
199.80 ± 60.9

69.67 ± 14.15 89.83 ± 8.77 69.20 ± 10.55

84.00 ± 10.58 79.33 ± 10.39 86.00 ± 14.99 77.50 ± 3.91

30.67 ± 2.89 31.83 ± 7.08 25.80 ± 6.38

35.33 ± 7.02 75.33 ± 34.60 70.00 ± 17.51 72.33 ± 29.92

46.86 ± 7.43 3.06 ± 0.18 4.39 ± 1.65
51.21 ± 7.89 2.63 ± 0.09 1.78 ± 0.43
39.32 ± 5.95 2.87 ± 0.33 4.88 ± 2.54

30.71 ± 2.88 3.55 ± 0.55 3.42 ± 2.60

31.97 ± 2.49 3.22 ± 0.40 4.00 ± 2.10

34.51 ± 4.06 4.10 ± 0.27 6.82 ± 0.46

40.05 ± 11.37 3.34 ± 0.24 4.85 ± 2.71

Connection according to US-PS 36 21 043 and US-PS 37 28 459. Weight percent of the additive in the food.

45

50

55

60

Table IV

Effect on food consumption and feed efficacy in rats (fed with a semi-purified basic diet supplemented with cholesterol)

Treatment " ¹ ) number
Rats Dose ² )
mg / kg /
Day Liver weight,
total g ± SD
100 g each
Body weight Body weight, g ± SD
beginning end 4th 208 ± 13 Close
rungsm,
ver
zehr, g Gross-
foulter
effective
samk. ^b ) No 6th 7.5 ± 0.6 3.6 ± 2 132 ± 5 208 ± 16 321 0.237 Clofibrate 0.2% 6th 198 10.4 ± 2.6 4.9 ± 1.2 135 ± 13th 208 ± 18 340 0.215 Cholestyramine, 3% 6th 2930 6.1 ± 0.5 2.9 ± 0.2 132 ± 336 0.226 Amides of Norfenflur-
amide (ex. 5) 6th 177 ± 15 0.1% 6th 91 5.9 + 0.5 3.4 ± 0.2 138 ± 9 190 ± 7 288 0.135 0.2% 6th 194 6.1 ± 0.3 3.2 ± 0.1 148 ± 12th 172 ± 11 328 0.128 0.5% 6th 466 5.7 ± 0.4 3.3 ± 0.1 148 ± 8th 192 ± 25 298 0.081 ö-Meihyibenzyl-
linoleamide ^c ) 0.2% 4th 199 7.1 ± 1.3 3.7 ± 0.2 137 ± 311 0.177

^a ) = based on% drugs in the diet and average daily feed consumption.

^b ) = gross weight increase / gross total food increase in g.

^c ) = connection according to US-PS 36 21 043 and US-PS 37 28 459.

^d ) = weight percent of the additive in food.

30 Table V

Effect on serum and liver lipids of rats (fed with a semi-purified basic diet with cholesterol)

Treatment ^c ) Lipids Serum, mg / 100 ml ± SD liver, mg / g ± SD Total Cholesterol Trigiyceride Total Cho'estcrin Triglyceride

no clofibrate, 0.2% cholestyramine, 3% amides of norfenfluramine (Ex. 5) 0.1% 0.2% 0.5%

ar-methylbenzyllinoleamide ^b ) 0.2%

635.67 ± 73.03 272.67 ± 44.19 430.33 + 119.37 ") 235.0O ± 68.27

253.83 ± 67.69 120.00 ± 23.22 25.37 + 6.51 12.45 ± 10.10 126.50 ± 47.21 ^a ) 93.83 ± 12.06 ^a ) 14.17 ± 9.10 ^a ) 7.83+ 2.22

242.83 ± 70.05 ^a ) 93.67 ± 17.50 ^a ) 180.00 + 82.19 76.67 ± 8.09 ^a ) 6.15 ± ^{1.37 a} ) 11.13 ± 4.11

375.67+ 39.1 l ^a ) 166.17 ± 44.53 ^a ) 176.00 ± 33.92 ^a ) 83.83+ 7.96 ")

266.33 ± 40.27 ^a ) 141.33 ± 28.08 ^a ) 184.67 ± 44.59 78.83 ± 12.83 ^a )

286.67 ± 58.03 ^a ) 163.00 ± 23.24 ^a ) 299.50 + £ 51.8. 75.33 ± 12.94 ^a )

453.25 + 9O.58 ^a ) 197.75 + 23.24 262.50 + 51.88 84.00 ± 14.26 ^a )

^a ) = ρ < 0.05 by Student's t-test.

^b ) = connection according to US-PS 36 21 043 and US-PS 37 28 459.

^c ) = weight percent of the additive in food.

4.80 ± 3.42 ^a ) 7.33+ 0.79

2.55 ± 2.65 ^a ) 6.50 ± 0.97

l, 50 ± 0.50 ^a ) 7.07 ± 0.86

8.33 ± 1.37 ^a ) 4.35 ± 2.88

10

Table VI

Effect of norfenfluramine linoleamide on serum and liver lipids in rats
(fed on a hypercholesteremic diet) (a) (b)

Treatment " ¹ ) serum lipids rag / 100 ml hepatic lipids mg / g

Cholesterol Triglycerides Phospholipids Cholesterol Triglycerides Phospholipids

None 454 + 121 105 ± 37 97.5 ± 32.6 57.32 ± 4.06 30.69 ± 14.66 23.98 ± 5.44

Linoleamide of 212 ± 30 ^c ) 156 ± 62 °) 105.4 ± 18.4 7.15 ± 0.73 ^c ) 5.59 ± 3.67 ^C ) 24.1 +8.75 norfenfluramine
(Ex. 1) 0.2%

^a ) = ± values are standard deviations of 10 animals per group.

^b ) = semi-purified diet supplemented with 1% cholesterol and 0.5% cholic acid at the expense of glucose.

^c ) = ρ < 0.05 determined according to Student's t-test

^d ) = weight percent of the additive in food.

15th

Table VU

Effect on serum lipoproteins of rats (fed on semi-purified diet supplemented with cholesterol)

Treatment ¹ -) number Percentage composition ³ ) LDL HDL albumin Tapes VLDL 45.94 2.52 13.93 no 6th 37.53 42.56 9.03 22.63 Clofibrate 0.2% 8th 25.76 35.07 16.15 31.77 Cholestyrumine 3 1 « 7th 17.00 Linoleamide of Norfenfiuramine - ;. 8sp. 5) 11.29 20.07 31.69 0.1% 6th 36.92 15.42 12.11 42.81 0.2% 6th 23.65 16.98 2.76 40.46 0.5% 4th 35.78 18.51 29.62 15.65 a-methylbenzyllinoleamide ^b ) 0.2% 7th 36.19

^a ) = VLDL = very low density lipoproteins; LDL = low density lipoproteins; HDL = lipoproteins where density,

determined with a Beckman-Gilford spectrophotometer.

^b ) = connection according to US-PS 36 21 043 and US-PS 37 28 459.

^c ) = weight percent of the additive in food.

20th

30 35 40

The LD ₅ "value of the amides of norfenfluramine, determined by a method according to Finney Statistical Methods in Biological Assay, Hafner Pub. Co., New York, 2nd edition (1964) was greater than 10,000 mg / kg, po. The KD ₅ "value of norfenfluramine was 137 mg / kg according to the same method. The therapeutic index of the amides of norfenfluramine appeared to be very favorable compared to norfenfluramine and no adverse reactions of the amides to serum glucose, microsomal enzyme, or monoamine oxidase were observed.

Synthesis of ^ ß-sterol and unsaponifiable substances

The effect of the mixed amides of Example 5 and the reference compounds in the./? -sterine synthesis was by a modified procedure according to Hill and Dvornik Arch. Biochem. Biophys. 114 (1966) 88 determined. Male Sprague-Dawley rats weighing 127-175 g freely displaced a semi-purified basic diet and water. The rats were placed in cages with raised wire floors in one Room that was kept at a temperature of 24 to 26 ° C, at a 12 hour day and 12 hour Night rhythm housed. The rhythm was reversed so that the night cycle corresponds to the working day to take advantage of the circadian rhythm, which promotes cholesterol synthesis. The test period was 5 days, and 20 ^ ci sodium acetate-l- ^ C (Sp. Act. 56 mei / m mol) administered intraperitoneally. Animals were sacrificed under carbon dioxide, and the distal 10 cm of the ilium as well the mcdianas and the left lobes of the liver were excised and homogenized. After that, the / i-Sterinc isolated as digitonide, and the radioactivity was measured in a "Packard Tricarb Liquid Scintillation Spectrometer" Model 3385 using 0.5% 2,5-diphenyloxazole and 8% naphthalene in one Solvent mixture of 40% toluene, 40% dioxane and 20% absolute ethanol determined.

Information on the effect of the mixed amides from Example 5 on the synthesis of non-saponifiable substances undjö sterols as determined by the digitonide method are made in Table 8. The results

show that at doses of 140 mg / kg, p. o., the amides of the invention reverse the sterol synthesis from control values 48% in the liver and 15% in the ilium decreased. A prior art compound, ar-methylbenzyllinoleamide, had at a dose of 140 mg / kg, p. o., no effect on cholesterol synthesis in either Tissues.

Table VIII

jff sterol synthesis.

Incorporation of acetate-l - '^ C in non-saponifiable and digitonin-precipitated sterols

link

Number of dose of tissue acetate-I- ^I4 C incorporated into rats mg / kg η mol / g tissue ± SD

^p '° "non-saponifiable digitoniol

No

Clofibrate

Cholestyramines

²⁵ Linoleic acid and mixed acid amides (Ex. 5) from Norfenfluramine

15th 140 liver 4.123 ± 1.718 3.028 ± 1.072 15th Ilium 7.841 ± 2.467 6.752 ± 1.248 10 660 liver 2.943 ± 1.707 1.588 ± 1.216 10 ilium 6.774 ± 1.447 6.098 ± 0.909 10 140 liver 14.869 ± 4.027 15.456 ± 2.710 Ilium 13.533 ± 3.904 12.085 ± 1.430 4th 140 liver 2.077 ± 1.246 !, 581 ± 1.158 4th Ilium 7.022 ± 2.125 5.731 ± 1.499 4th liver 4.533 ± 3.131 3.882 ± 2.387 4th Ilium 7.648 ± 1.664 5.947 ± 0.654

ff-MethylbenzyllinoIeamide ")

') Connection according to US-PS 36 21 043 and 37 28 459.

Formulation and administration

The cholesterol lowering anti-obesity agents according to the invention can be administered orally; usually the amount administered is 0.5 to 10 g per day, preferably 0.25 to 2.5 g per day, and the Administration can continue for several months. Anyone can use the cholesterol-lowering agent in a suitable form conventional for oral administration. So it can be enclosed in a capsule be or in liquid form, e.g. B. as a slurry, in tablet or powder form. In the preparation of The agent in these various forms can carry the active compound with a liquid carrier such as an edible oil. Mixtures of two or more of the agents according to the invention can can be used in any of the administration forms described above.

Tablet manufacturing

A granulation is made from

Lactose

strength

Water, sufficient door the granulation

The granulation is dried and sieved.

Amide of norfenfluramine (structural formula I) lactose granulation (as above) Magnesium stearate

Parts by weight

74 26

Ge * .- parts

100

145

are mixed and compressed into the form of tablets weighing 250 mg and 100 g of the. active ingredient 60 per unit dose included.

Administration in the form of a slurry Ks, an elixir is made, which per liter contains:

Linoleamide from Norfenfluramine (structural formula I) 100.0 g 5

Ethyl alcohol 150.0 ml

Glycerin 350.0 ml

Sorbitol (70% solution) 350.0 ml

Benzoic acid 1.0 g

Sodium cacharine 0.3 g 10

Colorant (FD and C Red No. 2) 0.02 g

artificial raspberry flavor 0.2 ml

Spiced vanilla 0.02 ml

distilled water, remainder to 1000 ml

The active ingredient of formula I is added to about two thirds of the ethanol, all of the glycerin and sorbitol are added and the mixture thoroughly triturated. The saccharin and the dye are dissolved in a small amount of water and dye dissolved in it. The aqueous solution is then added to the alcohol solution, and the water residue necessary for a volume of one liter is added; after mixing and filtering, an elixir containing 20 mg of active ingredient / ml is obtained. A 20 A unit of a dose of 15 ml (one tablespoon) contains 150 mg of the active ingredient.

It can all linoleamides of the invention or mixtures thereof for the preparation of the tablet or Elixir form can be used. The active ingredient can suitably be within the range of 50 to 400 mg and preferably between 50 and 200 mg per unit dose can be varied. Also can other therapeutic agents can be added to these formulations if desired. 25th

The powder form is made with the addition of various conditioning agents such as starches, gums, etc. made for mixing with food for humans and for animals.

13th

Claims (1)

Patent claims: 1. Fatty acid amides of ff-MethyI-m- (trifluoromethyl) -phenäthylamin (Norfenfluramine) the general formula CH ₃ O I Il -CH ₂ -CH-NCR (I)