WO1999025682A1

WO1999025682A1 - Elevation of hdl cholesterol by 4-[(aminothioxomethyl) -hydrazono] -n-(substituted) -4-arylbutanamides

Info

Publication number: WO1999025682A1
Application number: PCT/US1998/024479
Authority: WO
Inventors: Thomas Joseph Commons
Original assignee: American Home Products Corp
Current assignee: Wyeth LLC
Priority date: 1997-11-17
Filing date: 1998-11-16
Publication date: 1999-05-27
Anticipated expiration: 2000-05-17
Also published as: AU1525999A

Abstract

Compounds of this invention increase plasma levels of high density lipoprotein or HDL, the 'good' cholesterol and as such may be useful for treating diseases such as atherosclerosis. These compounds are represented by formula (A), wherein: R?1, R2, and R3¿ are independently hydrogen, C¿1?-C6 alkyl, phenyl or -(CH2)1-6 phenyl where phenyl is optionally substituted by halogen, cyano, nitro, C1-C6 alkyl, C1-C6 alkoxy, trifluoromethyl, C1-C6 alkoxycarbonyl, -CO2H or OH; R?4 and R5¿ are independently hydrogen, C¿1?-C10 alkyl, C3-C8 cycloalkyl, -(CH2)0-6Ar?1¿ where Ar1 is phenyl, naphthyl, furanyl, pyridinyl or thenyl and Ar1 can be optionally substituted by halogen, cyano, nitro, C¿1?-C6 alkyl, phenyl, C1-C6 alkoxy, phenoxy, trifluoromethyl, C1-C6 alkoxycarbonyl, -CO2H or OH, or R?4 and R5¿ together with the nitrogen to which R?4 and R5¿ are attached from a ring containing 4-7 carbon atoms; and Ar is phenyl, naphthyl, furanyl, pyridinyl or thienyl which may be optionally substituted by halogen, cyano, nitro, C¿1?-C6 alkyl, C3-C6 cycloalkyl, phenyl, C1-C6 alkoxy, phenoxy, trifluoromethyl, C1-C6 alkoxycarbonyl, -CO2H or OH.

Description

Elevation of HDL Cholesterol by 4-[(Aminothioxomethyl)- Hydrazono]-N-(Substituted)-4-Arylbutanamides

Field of Invention

This invention relates to compounds useful in elevating high density lipoprotein, the "good" cholesterol. Compounds of this invention increase plasma levels of HDL in a cholesterol fed rat model and as such these compounds may be useful for treating diseases such as atherosclerosis.

Background of the Invention

It is widely beleived that HDL is a "protective" lipoprotein [Gloria Lena Vega and Scott Grundy, Current Opinion in Lipidology, 7, 209-216 (1996)] and that increasing plasma levels of HDL may offer a direct protection against the development of atherosclerosis. Numerous studies have demonstrated that both the risk of coronary heart disease (CHD) in humans and the severity of experimental atherosclerosis in animals are inversely correlated with serum HDL cholesterol (HDL-C) concentrations (Russ et al., Am. J. Med.. ϋ (1951) 480-493; Gofman et al, Circulation. 34 (1966) 679-697; Miller and Miller, Lancet. I (1975) 16-19; Gordon et al., Circulation. 79 (1989) 8-15; Stampfer et al., N. Engl. J. Med.. 325 (1991) 373-381; Badimon et al., Lab. Invest.. 60 (1989) 455-461). Atherosclerosis is the process of accumulation of cholesterol within the arterial wall which results in the occlusion, or stenosis, of coronary and cerebral arterial vessels and subsequent myocardial infarction and stroke. Angiographical studies have shown that elevated levels of some HDL particles in humans appears to be correlated to a decreased number of sites of stenosis in the coronary arteries of humans (Miller et al., Br. Med. J.. 282 (1981) 1741-1744).

There are several mechanisms by which HDL may protect against the progression of atherosclerosis. Studies in vitro have shown that HDL is capable of removing cholesterol from cells (Picardo et al., Arteriosclerosis. 6 (1986) 434-441). Data of this nature suggest that one antiatherogenic property of HDL may lie in its ability to deplete tissues of excess free cholesterol and eventually lead to the delivery of this cholesterol to the liver (Glomset, J. Lipid Res.. 9 (1968) 155-167). This has been supported by experiments showing efficient transfer of cholesterol from HDL to the liver (Glass et al., Circulation. 66 (Suppl. II) (1982) 102; MacKinnon et al., J. Biol. Chem.. 261 (1986) 2548-2552). In addition, HDL may serve as a reservoir in the circulation for apoproteins necessary for the rapid metabolism of triglyceride-rich lipoproteins (Grow and Fried, J. Biol. Chem.. 253 (1978) 8034-8041; Lagocki and Scanu, J. Biol. Chem.. 255 (1980) 3701-3706; Schaefer et al., J. Lipid Res.. 23 (1982) 1259-1273). Accordingly, agents which increase HDL cholesterol concentrations are useful as anti-atherosclerotic agents, particularly in the treatment of dyslipoproteinemias and coronary artery disease.

Takagi et al., US 5,608,109, discloses agricultural and horticultural insecticidal compounds according to formula A below where Ar and R² are both optionally substituted phenyl, R¹ and R³ are independently hydrogen, alkyl, alkenyl or alkynyl, and R⁴ and R⁵ are independently hydrogen or alkyl.

BRIEF DESCRIPTION OF THE INVENTION

The compounds of this invention which elevate plasma levels of HDL cholesterol have the general structure A

A

wherein:

R¹, R², and R³ are independently hydrogen, -C_ό alkyl, phenyl or -(CH₂)ι- phenyl where phenyl is optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, -C alkyl, -C_ό alkoxy, trifluoromethyl, -C₆ alkoxycarbonyl, -CO₂H or OH;

R⁴ and R⁵ are independently hydrogen, - o alkyl, C₃-C0, cycloalkyl, -(CT^o-_όAr¹ where Ar¹ is phenyl, naphthyl, furanyl, pyridinyl or thenyl and Ar¹ can be optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, -C alkyl, phenyl, Cι-C₆ alkoxy, phenoxy, trifluoromethyl, Cι-C alkoxycarbonyl, -CO₂H or OH, or R⁴ and R⁵ together with the nitrogen to which R⁴ and R⁵ are attached form a ring containing 4-7 carbon atoms;

and Ar is phenyl, naphthyl, furanyl, pyridinyl or thienyl which may be optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, Cι-C₆ alkyl, C₃-C₆ cycloalkyl, phenyl, C C alkoxy, phenoxy, trifluoromethyl, - alkoxycarbonyl, -CO₂H or OH.

When used herein as a group or part of a group the term alkyl may be straight or branched, including methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, n-pentyl, n-hexyl and n-heptyl. When used herein the term alkenyl may be straight or branched, including, ethenyl, 1-propenyl and 2-propenyl. When used herein the term alkynyl may be straight or branched, including, ethynyl, 1-propynyl and 2-propynyl.

When used herein the term cycloalkyl may be saturated or unstaurated, including, cyclopropyl, cyclobutyl, cyclopenyl, cyclohexyl, cyclopropenyl, cyclobutenyl and cyclopentenyl. When used herein the term halogen may be chlorine, bromin3, fluorine or iodine.

R¹, R², and R³ are preferably selected from hydrogen and Cι-C alkyl, more preferably from hydrogen and methyl. R⁴ and R⁵ are preferably selected from hydrogen, - o alkyl, C₃-C₈ cycloalkyl, and -(CH₂)o-₆Ar^l, more preferably from hydrogen, methyl, i-propyl, n-butyl, 1,5-dimethylhexyl, cyclohexyl, and benzyl. Ar is preferably optionally substituted phenyl.

The compounds are tested in vivo in rats fed cholesterol-augmented rodent chow for 8 days according to the test protocol and blood from the rats analyzed for HDL cholesterol.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of this invention are prepared by reacting 4-oxo-4-arylbutyric acid amides with an appropriately substituted thiosemicarbazide according to Scheme I. The intermediate 4-oxo-4-arylbutyric acid amides are conveniently prepared by the routes shown in Scheme II by reacting an amine of the formula HNR⁴R⁵ with either a 4-aryl-4-oxobutyric acid or a γ-aryl-γ-butyrolactone. Specific examples are given in the Experimental Section. Scheme I: Preparation of title compounds.

Scheme LI: Preparation of intermediate 4-oxo-4-phenylbutyric acid amides.

(1) Route A to Ketoamide i

(2) Route B to Ketoamide 1

R⁴R⁵NH PyHClCrO₃

Solvent CH₂C1₂

1

The following examples are included for illustrative purposes only and are not to be construed as limiting to this disclosure in any way. Those skilled in the art of synthetic organic chemistry may be aware of other methods of preparing compounds of this invention. The starting materials or intermediates are available commercially or can be prepared by following standard literature procedures. Example 1

2-(4-Butylamino-4-oxo-l-phenyl-butylidene)-hydrazinothiocarboxamide (A) 1 ,3-Diisopropylcarbodiimide (22.0 ml, 0.14 moles) in 200 ml of methylene chloride was added under nitrogen dropwise over 45 minutes to a solution of 3- benzoylpropionic acid (25.00g, 0.14 moles), 4-dimethylaminopyridine (17.16g, 0.14 moles) and butylamine (13.9 ml, 0.14 moles) in 500 ml of methylene chloride at ice bath temperature. After the addition the reaction was stirred at ice bath temperature for 1 hour. The ice bath was removed and the reaction stirred for 18 hours. The solid present was removed by filtration and discarded. The filtrate was extracted with 1 N HCl, 5% NaHCO₃, dried (MgSO₄) and the solvent removed under reduced pressure to give 35.84g of a yellow solid. Recrystallization of the solid from isopropyl alcohol gave N-butyl-4-oxo-4-phenyl-butyramide (15.99g, 49%) as a light yellow solid, mp 78-80°C.

Elemental Analysis for C₁₄H₁₉NO₂ Calc'd: C, 72.07; H, 8.21; N, 6.00 Found: C, 72.42; H, 8.20; N, 6.07

(B) Thiosemicarbazide ( 1.95g, 21.4 mmol) was added to a solution of N-butyl-4- oxo-4-phenyl-butyramide (5.0g, 21.4 mmol), prepared in the previous step, in 75 ml of methanol plus 5.8 ml of 1 N HCl plus 5.8 ml of water and the reaction stirred at room temperature for 22 hours. The reaction was concentrated under reduced pressure at which time a solid formed. The solid was collected by filtration and dried to give 4.49g of an off-white solid. Recrystallization of the solid from isopropyl alcohol gave 3.16g (48%) of the title compound as a white solid, mp 64-68°C.

Elemental Analysis for C₁₅H₂₂N₄OS'0.8 C₃H₈O Calc'd: C, 58.95; H, 8.07; N, 15.80

Found: C, 58.93; H, 8.25; N, 15.45 Example 2

4-[(Aminothioxomethyl)hydrazono]-N-(l-methylethyl)-4-phenyl- butanamide (A) In the same manner as described in step (A) of Example 1 ,and replacing butylamine with isopropylamine, N-(l-methylethyl)-4-oxo-4-phenyl-butyramide (10.24g, 33%) was obtained as a white solid, mp 118-121°C.

Elemental Analysis for C₁₃H_]7NO₂ Calc'd: C, 71.21; H, 7.81; N, 6.39

Found: C, 71.14; H, 7.83; N, 6.36

(B) Thiosemicarbazide (2.08g, 22.8 mmol) was added to a solution of N-(l- methylethyl)-4-oxo-4-phenyl-butyramide (5.0g, 22.8 mmol), prepared in the previous step, in 80 ml of methanol plus 6.2 ml of 1 N HCl plus 6.2 ml of water and the reaction stirred at room temperature for 2 days. An additional 1.04g (11.4 mmol) of thiosemicarbazide was added and the reaction stirred at room temperature for 24 hours. The reaction was cooled in an ice bath and a white solid precipitated. The solid was collected by filtration and dried. Recrystallization of the solid from isopropyl alcohol gave the title compound (4.73g, 53%) as a white solid, mp 66-71°C.

Elemental Analysis for C₁₄H₂₀N₄OS^«1.6 C₃H₈O Calc'd: C, 58.12; H, 8.51; N, 14.42 Found: C, 57.06; H, 8.58; N, 13.80

Example 3

4-[(Aminothioxomethyl)-hydrazono]-4-phenylbutanamide

(A) A mixture of γ-phenyl-γ-butyrolactone (10.30g, 63.5 mmol) and an excess of ammonia was stirred under nitrogen and a dry ice trap for 8 hours. The dry ice trap was removed and after evaporation of the ammonia 11.27g of a tan solid remained. Recrystallization of the solid from ethyl acetate-hexane gave 4-hydroxy-4-phenyl- butyramide (8.56g, 75%) as a white solid, mp 85-87°C.

Elemental Analysis for C₁₀H₁₃NO₂

Calc'd: C, 67.02; H, 7.31; N, 7.82 Found: C, 67.27; H, 7.25; N, 7.84 (B) A mixture of 4-hydroxy-4-phenyl-butyramide (4.02g, 22.4 mmol), prepared in the previous step, and pyridinium chlorochromate (7.26g, 33.7 mmol) in 500 ml of methylene chloride was stirred at room temperature for 2 hours. The reaction was poured onto 400g of silica gel (230-400 mesh) and the material eluted with ethyl acetate. Isolation of the major component gave 1.92g of a purple solid. Recrystallization of the solid from ethyl acetate gave 4-oxo-4-phenyl-butyramide (1.24g, 31%) as a green solid, mp 122-124°C.

Elemental Analysis for CioH] iNO₂

Calc'd: C, 67.78; H, 6.26; N, 7.90 Found: C, 67.34; H, 6.15; N, 7.75

(C) Thiosemicarbazide (1.38g, 15.1 mmol) was added to a solution of 4-oxo-4- phenyl-butyramide (1.63g, 9.22 mmol), prepared in the previous step, in 50 ml of methanol plus 2.5 ml of 1 N HCl plus 2.5 ml of water and the reaction stirred at room temperature for 28 hours. The solid formed was collected by filtration and dried to give 1.73g of a purple solid. Recrystallization of the solid from methanol gave the title compound (1.34g, 58%) as a purple solid, mp 186-188°C.

Elemental Analysis for C_πH₁₄N₄OS Calc'd: C, 52.78; H, 5.64; N, 22.38 Found: C, 52.75; H, 5.66; N, 22.41

Example 4

4-[(Aminothioxomethyl)-hydrazono]-N-benzyl-4-phenylbutanamide

(A) A solution of γ-phenyl-γ-butyrolactone (5.17g, 31.8 mmol) and benzylamine (3.48 ml, 31.8 mmol) in 200 ml of benzene was refluxed under a nitrogen atmosphere for 20 hours. An additional 1.75 ml (16.0 mmol) of benzylamine was added and the solution refluxed for 24 hours. When the reaction was partitioned with 1 N HCl a white solid precipitated. The solid was collected by filtration and dried to give N- benzyl-4-hydroxy-4-phenyl-butyramide (6.12g, 72%) as a white solid, mp 92-94°C.

Elemental Analysis for C i.₇H _{( 9}NO₂

Calc'd: C, 75.81; H, 7.11 ; N, 5.20 Found: C, 75.64; H, 7.08; N, 5.08 (B) A mixture of N-benzyl-4-hydroxy-4-phenyl-butyramide (5.58g, 20.7 mmol), prepared in the previous step, and pyridinium chlorochromate (6.69g, 31.0 mmol) in 600 ml of methylene chloride was stirred at room temperature for 1.5 hours. The reaction was poured onto 300g of silica gel (230-400 mesh) and the material eluted with methylene chloride-ethyl acetate. Isolation of the major component gave 4.68g (85%) of a green solid. Recrystallization of the solid from ethyl acetate gave N-benzyl-4-oxo- 4-phenyl-butyramide as a white solid, mp 110-112°C.

Elemental Analysis for C ₁₇H ₁₇NO₂

Calc'd: C, 76.38; H, 6.41; N, 5.24 Found: C, 76.33; H, 6.14; N, 5.26

(C) Thiosemicarbazide (2.30g, 25.2 mmol) was added to a solution of N-benzyl-4- oxo-4-phenyl-butyramide (4.12g, 15.4 mmol), prepared in the previous step, in 100 ml of methanol plus 4.2 ml of 1 N HCl plus 4.2 ml of water and the reaction stirred at room temperature for 20 hours. The reaction was cooled in an ice bath and a white solid precipitated. The solid was collected by filtration and dried to give 4.61g of a white solid. Recrystallization of the solid from ethyl acetate-methanol gave the title compound (2.05g, 39%) as a white solid, mp 162-164°C.

Elemental Analysis for C_l8H₂₀N₄OS Calc'd: C, 63.50; H, 5.92; N, 16.46 Found: C, 63.48; H, 5.77; N, 16.60

Example 5

4-[(Aminothioxomethyl)-hydrazono]-N-methyl-4-phenylbutanamide

(A) A solution of γ-phenyl-γ-butyrolactone (5.17g, 31.9 mmol) in 40 ml of a 2 molar solution of methylamine in THF was stirred under nitrogen at room temperature for 23 hours. The solvent was removed under reduced pressure to give 6.15g of a yellow solid. Recrystallization of the solid from ethyl acetate-hexane gave 4-hydroxy- N-methyl-4-phenyl-butyramide (4.8 lg, 78%) as an off-white solid, mp 67-69°C.

Elemental Analysis for ₁H₁₅NO₂

Calc'd: C, 68.37; H, 7.82; N, 7.25 Found: C, 68.60; H, 7.98; N, 7.32 (B) A mixture of 4-hydroxy-N-methyl-4-phenyl-butyramide (4.50g, 23.3 mmol), prepared in the previous step, and pyridinium chlorochromate (7.54g, 35.0 mmol) in 200 ml of methylene chloride was stirred at room temperature for 2.5 hours. The reaction was poured onto 400g of silica gel (230-400 mesh) and the material eluted with ethyl acetate-methylene chloride and then ethyl acetate. Isolation of the major component gave N-methyl-4-oxo-4-phenyl-butyramide (3.65g, 82%) as an off-white solid, mp 80-82°C.

Elemental Analysis for C_nH₁₃NO₂ Calc'd: C, 69.09; H, 6.85; N, 7.32

Found: C, 69.08; H, 6.87; N, 7.27

(C) Thiosemicarbazide (2.60g, 28.5 mmol) was added to a solution of N-methyl-4- oxo-4-phenyl-butyramide (3.33g, 17.4 mmol), prepared in the previous step, in 60 ml of methanol plus 4.7 ml of 1 N HCl plus 4.7 ml of water and the reaction stirred at room temperature for 22 hours. The reaction was submerged in an ice bath and a solid precipitated. The solid was collected by filtration and dried to give 4.55g of a white solid. The solid was triturated with water and then dried to give the title compound (4.10g, 89%) as a white solid, mp 205-207°C.

Elemental Analysis for C₁₂H₁₆N₄OS Calc'd: C, 54.52; H, 6.10; N, 21.19 Found: C, 54.55; H, 6.02; N, 21.20

Example 6

4-[(AminothioxomethyI)-hydrazono]-N-cyclohexyl-4-phenylbutanamide

(A) A solution of γ-phenyl-γ-butyrolactone (5.16g, 31.8 mmol) and cyclohexylamine (7.3 ml, 63.8 mmol) in 200 ml of benzene was refluxed under a nitrogen atmosphere for 3 days. The reaction was extracted with 1 N HCl, dried (MgSO₄) and the solvent removed under reduced pressure to give 7.07g of a brown solid. Recrystallization of the solid from ethyl acetate-hexane gave N-cyclohexyl-4- hydroxy-4-phenyl-butyramide (4.22g, 51%) as a white solid, mp 91-93°C.

Elemental Analysis for C₁₆H₂₃NO₂

Calc'd: C, 73.53; H, 8.87; N, 5.36 Found: C, 73.48; H, 9.12; N, 5.48 (B) A mixture of N-cyclohexyl-4-hydroxy-4-phenyl-butyramide (3.98g, 15.2 mmol), prepared in the previous step, and pyridinium chlorochromate (4.94g, 22.9 mmol) in 200 ml of methylene chloride was stirred at room temperature for 2.5 hours. The reaction was poured onto 400g of silica gel (230-400 mesh) and the material eluted with methylene chloride-ethyl acetate. Isolation of the major component gave N- cyclohexyl-4-oxo-4-phenyl-butyramide (3.28g, 83%) as a light green solid, mp 109- 111°C.

Elemental Analysis for C i.₆H₂1 NO₂

Calc'd: C, 74.10; H, 8.16; N, 5.40 Found: C, 73.64; H, 8.11; N, 5.41

(C) Thiosemicarbazide (1.75g, 19.2 mmol) was added to a solution of N- cyclohexyl-4-oxo-4-phenyl-butyramide (3.30g, 11.7 mmol), prepared in the previous step, in 40 ml of methanol plus 3.2 ml of 1 N HCl plus 3.2 ml of water and the reaction stirred at room temperature for 21 hours. The solid formed was removed by filtration and dried to give 2.69g of a white solid. The filtrate was concentrated under reduced pressure to remove the methanol. The residue was partitioned between methylene chloride and water. The organic layer was separated, washed multiple times with water, dried (MgSO₄) and the solvent removed under reduced pressure to give 1.1 Og of a white foam. The initial solid filtered from the reaction was dissolved in methylene chloride, washed multiple times with water, dried (MgSO₄) and the solvent removed under reduced pressure to give a white foam. Both materials were combined and crystallized from methylene chloride-ethyl acetate to give the title compound (3.3 lg, 69%) as a white solid, mp 89-92°C.

Elemental Analysis for C₁₇H₂₄N₄OS^»0.9 C₄H₈O₂ Calc'd: C, 60.09; H, 7.64; H; 13.61 Found; C, 59.98; H, 7.67; N, 13.43

Example 7

4-[(Aminothioxomethyl)hydrazono]-N,N-dimethyl-4-phenylbutanamide (A) A solution of γ-phenyl-γ-butyrolactone (4.84g, 29.8 mmol) in 200 ml of a 2 molar solution of dimethylamine in THF was stirred under a nitrogen atmosphere at room temperature for 2 days. The solvent was removed under reduced pressure to give 6.26g of 4-hydroxy-N,N-dimethyl-4-phenyl-butyramide as a light brown oil, MS [M+H]⁺ m/e 208.

Elemental Analysis for C₁₂H_I7NO₂ Calc'd: C, 69.54; H, 8.27; N, 6.76

Found: C, 68.13; H, 8.28; N, 6.65

(B) In the same manner as described in step (B) of Example 6, 4-oxo-N,N- dimethyl-4-phenyl-butyramide (4.08g, 72%) was obtained as an off-white solid, mp 56-58°C.

Elemental Analysis for C₁₂H₁₅NO₂ Calc'd: C, 70.22; H, 7.37; N, 6.82 Found; C, 70.08; H, 7.40; N, 6.95

(C) Thiosemicarbazide (2.75g, 30.2 mmol) was added to a solution of 4-oxo-N,N- dimethyl-4-phenyl-butyramide (3.80g, 18.5 mmol), prepared in the previous step, in 65 ml of methanol plus 5 ml of 1 N HCl plus 5 ml of water and the reaction stirred at room temperature for 28 hours. The solid present was collected by filtration to give 4.76g of a white solid. The solid was triturated with water and then dried to give the title compound (4.53g, 88%) as a white solid, mp 173-175°C.

Elemental Analysis for Cι₃H₁₈N₄OS Calc'd: C, 56.09; H, 6.52; N, 20.13 Found: C, 55.64; H, 6.37; N, 19.78

Example 8

l-[4-[(AminothioxomethyI)hydrazono]-l-oxo-4-phenylbutyl]piperidine (A) In the same manner as described in step (A) of Example 6, and replacing cyclohexylamine with piperidine, 4-hydroxy-4-phenyl-l-piperidin-l-yl-butan-l-one (4.86g, 62%) was obtained as an off-white solid, mp 42-44°C.

Elemental Analysis for C₁₅H₂₁NO₂ Calc'd: C, 72.84; H, 8.56; N, 5.66

Found: C, 72.83; H, 8.82; N, 5.63 (B) A mixture of 4-hydroxy-4-phenyl-l-piperidin-l-yl-butan-l-one (4.00g, 16.2 mmol), prepared in the previous step, and pyridinium chlorochromate (5.23g, 24.3 mmol) in 150 ml of methylene chloride was stirred at room temperature for 2.25 hours. The reaction was poured onto 400g of silica gel (230-400 mesh) and the material eluted with methylene chloride-ethyl acetate. Isolation of the major component gave 4-oxo-4- phenyl-1-piperidin-l-yl-butan-l-one (3.66g, 92%) as a green solid. Recrystallization of a portion of this solid from ethyl acetate-hexane gave an analytically pure sample, mp 52-54°C.

Elemental Analysis for C₁₅H₁₉NO₂

Calc'd: C, 73.44; H, 7.81; N, 5.71 Found: C, 73.55; H, 7.95; N, 5.70

(C) In the same manner as described in step (C) of Example 7, the title compound (3.90g, 91%) was obtained as a white solid, mp 181-183°C.

Elemental Analysis for Cj₆H₂₂N₄OS Calc'd: C, 60.35; H, 6.96; N, 17.59 Found: C, 60.14; H, 7.06; N, 17.58

Example 9

4-[(Aminothioxomethyl)-hydrazono]-N-(l,5-dimethylhexyI)-4-phenyl- butanamide (A) A solution of γ-phenyl-γ-butyrolactone (5.02g, 30.9 mmol) and 1,5- dimethylhexylamine (10.4 ml, 61.7 mmol) in 200 ml of benzene was refluxed under a nitrogen atmosphere for 40 hours. An additional 10.4 ml (61.7 mmol) of 1,5- dimethylhexylamine was added and the reaction refluxed for 26 hours. The reaction was extracted with 1 N HCl, dried (MgSO₄) and the solvent removed under reduced pressure to give to give 8.26g of a light brown solid. Recrystallization of the solid from ethyl acetate-hexane gave N-(l,5-dimethyl-hexyl)-4-hydroxy-4-phenyl- butyramide (3.72g, 41%) as a white solid, mp 72-89°C.

Elemental Analysis for C₁₈H₂₉NO₂ Calc'd: C, 74.18; H, 10.03; N, 4.81

Found: C, 74.50; H, 10.34; N, 4.79 (B) A mixture of N-( 1 ,5-dimethyl-hexyl)-4-hydroxy-4-phenyl-butyramide (3.50g, 12.0 mmol), prepared in the previous step, and pyridinium chlorochromate (3.89g, 18.0 mmol) in 150 ml of methylene chloride was stirred at room temperature for 1.25 hours. The reaction was poured onto 400g of silica gel (230-400 mesh) and the material eluted with methylene chloride-ethyl acetate. Isolation of the major component gave N-(l,5-dimethyl-hexyl)-4-oxo-4-phenyl-butyramide (2.7 lg, 78%) as a light green solid, mp 84-87°C.

Elemental Analysis for Cι₈H₂₇NO₂ Calc'd: C, 74.70; H, 9.40; N, 4.84

Found: C, 74.38; H, 9.65; N, 4.89

(C) Thiosemicarbazide (1.32g, 14.5 mmol) was added to a solution of N-(l,5- dimethyl-hexyl)-4-oxo-4-phenyl-butyramide (2.54g, 87.9 mmol), prepared in the previous step, in 30 ml of methanol plus 2.4 ml of 1 N HCl plus 2.4 ml of water and the reaction stirred at room temperature for 48 hours. The reaction was concentrated under reduced pressure to remove the methanol. The residue was partitioned between methylene chloride and water. The organic layer was separated, washed multiple times with water, dried (MgSO₄) and the solvent removed under reduced pressure to give 2.95g of an off-white solid foam. Purification of this foam by chromatography on silica gel (230-400 mesh) using 3:1 hexane-ethyl acetate and then 3:1 ethyl acetate- methylene chloride as the eluents gave the title compound (2.63g, 82%) as a white foam, mp 58-65°C.

Elemental Analysis for C₁₉H₃₀N₄OS

Calc'd: C, 62.95; H, 8.34; N, 15.45 Found: C, 62.53; H, 8.47; N, 15.35

PHARMACOLOGY

In Vivo Assay: Male Sprague-Dawley rats weighing 200-225 g are housed two per cage and fed Purina Rodent Chow Special Mix 5001-S supplemented with 0.25% cholic acid and 1.0% cholesterol and water ad libitum for 8 days. Each test substance is administered to a group of six rats fed the same diet with the test diet mixed in as 0.005 - 0.1 % of the total diet. Body weight and food consumption are recorded prior to diet administration and at termination. Typical doses of the test substances are 5 - 100 mg/kg/day. At termination, blood is collected from anesthetized rats and the serum is separated by centrifugation. Total serum cholesterol is assayed using the Sigma Diagnostics enzymatic kit for the determination of cholesterol, Procedure No. 352, modified for use with ninety-six well microtiter plates. After reconstitution with water the reagent contains 300 U/I cholesterol oxidase, 100 U/I horse radish peroxidase, 0.3 mmoles/1 of 4-aminoantipyrine and 30.0 mmoles/1 of p-hydroxybenzenesulfonate in a pH 6.5 buffer. In the reaction cholesterol is oxidized to produce hydrogen peroxide which is used to form a quinoneimine dye. The concentration of dye formed is measured spectrophotometrically by absorbance at 490 nm after incubation at 25 °C for 30 minutes. The concentration of cholesterol was determined for each serum sample relative to a commercial standard from Sigma.

HDL cholesterol concentrations in serum are determined by separation of lipoprotein classes by fast protein liquid chromatography (FPLC) by a modification of the method of Kieft et al., J. Lipid Res., 32 (1991) 859-866. 25 μl of serum is injected onto Superose 12 and Superose 6 (Pharmacia), in series, with a column buffer of 0.05 M Tris (2-amino-2-hydroxymethyl-l,3-propanediol) and 0.15 M sodium chloride at a flow rate of 0.5 ml/min. The eluted sample is mixed on line with Boehringer- Mannheim cholesterol reagent pumped at 0.2 ml/min. The combined eluents are mixed and incubated on line through a knitted coil (Applied Biosciences) maintained at a temperature of 45° C. The eluent is monitored by measuring absorbance at 490 nm and gives a continuous absorbance signal proportional to the cholesterol concentration. The relative concentration of each lipoprotein class is calculated as the per cent of total absorbance. HDL cholesterol concentration, in serum, is calculated as the per cent of total cholesterol as determined by FPLC multiplied by the total serum cholesterol concentration. TABLE I

Cholesterol Fed Rat

Example % Increase in HDL (Dose)

Example 1 56 % (100 mg/kg) Example 2 61.7 % (100 mg/kg) Example 3 36.4 % (50 mg/kg) Example 4 49.5 % (50 mg/kg) Example 5 43.3 % (100 mg/kg) Example 6 28.1 % (82 mg/kg) Example 7 56.2 % (50 mg/kg) Example 8 52.8 % (50 mg/kg) Example 9 40.4 % (50 mg/kg)

PHARMACEUTICAL COMPOSITION

Compounds of this invention may be administered neat or with a pharmaceutical carrier to a patient in need thereof. The pharmaceutical carrier may be solid or liquid.

Applicable solid carriers can include one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrating agents or an encapsulating material. In powders, the carrier is a finely divided solid which is in admixture with the finely divided active ingredient. In tablets, the active ingredient is mixed with a carrier having the necessary compression properties In suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99% of the active ingredient. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins.

Liquid carriers may be used in preparing solutions, suspensions, emulsions, syrups and elixirs. The active ingredient of this invention can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fat. The liquid carrier can contain other suitable pharmaceutical additives such a solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmo-regulators. Suitable examples of liquid carriers for oral and parenteral administration include water (particularly containing additives as above, e.g., cellulose derivatives, preferable sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g., glycols) and their derivatives, and oils (e.g., fractionated coconut oil and arachis oil). For parenteral administration the carrier can also be an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are used in sterile liquid form compositions for parenteral administration.

Liquid pharmaceutical compositions which are sterile solutions or suspensions can be utilized by, for example, intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions can also be administered intravenously. Oral administration may be either liquid or solid composition form. The compounds of this invention may be administered rectally in the form of a conventional suppository. For administration by intranasal or intrabronchial inhalation or insufflation, the compounds of this invention may be formulated into an aqueous or partrially aqueous solution, which can then be utilized in the form of an aerosol. The compounds of this invention may also be administered transdermally through the use of a transdermal patch containing the active compound and a carrier that is inert to the active compound, is non-toxic to the skin, and allows delivery of the agent for systemic absorption into the blood stream via the skin. The carrier may take any number of forms such as creams and ointments, pastes, gels, and occlusive devices. The creams and ointments may be viscous liquid or semi-solid emulsions of either the oil in water or water in oil type. Pastes comprised of absorptive powders dispersed in petroleum or hydrophilic petroleum containing the active ingredient may also be suitable. A variety of occlusive devices may be used to realease the active ingredient into the blood stream such as a semipermeable membrane covering a reservoir containing the active ingredient with or without a carrier, or a matrix containing the active ingredient. Other occlusive devices are known in the literature.

The dosage to be used in the treatment of a specific patient suffering from high density lipoprotein insufficiency must be subjectively determined by the attending physician. The variables involved include the severity of the dysfunction, and the size, age, and response pattern of the patient.. Treatment will generally be initiated with small dosages less than the optimum dose of the compound. Thereafter the dosage is increased until the optimum effect under the circumstances is reached. Precise dosages for oral or parenteral administration will be determined by the administering physician based on experience with the individual subject treated and standard medical principles.

Preferably the pharmaceutical composition is in unit dosage form, e.g., as tablets or capsules. In such form, the composition is sub-divided in unit doses containing appropriate quantities of the active ingredient; the unit dosage form can be packaged compositions, for example packed powders, vials, ampoules, prefilled syringes or sachets containing liquids. The unit dosage form can be, for example, a capsule or tablet itself, or it can be the appropriate number of any such compositions in package form.

Claims

What is claimed:

1. A compound according to the formula:

A wherein:

R¹, R², and R³ are independently hydrogen, -C alkyl, phenyl or -(CT^ -₆ phenyl where phenyl is optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, Cι-C alkyl, Cj-C alkoxy, trifluoromethyl, -C alkoxycarbonyl, -CO₂H or OH;

R⁴ and R⁵ are independently hydrogen, -Cio alkyl, C₃-C₈ cycloalkyl, -(CH₂)o-₆Ar¹ where Ar¹ is phenyl, naphthyl, furanyl, pyridinyl or thenyl and Ar¹ can be optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, -Cβ alkyl, phenyl, -C₆ alkoxy, phenoxy, trifluoromethyl, -C alkoxycarbonyl, -CO₂H or OH, or R⁴ and R⁵ together with the nitrogen to which R⁴ and R⁵ are attached form a ring containing 4-7 carbon atoms;

and Ar is phenyl, naphthyl, furanyl, pyridinyl or thienyl which may be optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, - alkyl, C₃-C₆ cycloalkyl, phenyl, C C₆ alkoxy, phenoxy, trifluoromethyl, -C₆ alkoxycarbonyl, -CO₂H or OH;

with the proviso that Ar and R² cannot simultaneously be optionally substituted phenyl when both R¹ and R³ are independently hydrogen or alkyl.

2. A compound according to claim 1 which is selected from:

2-(4-butylamino-4-oxo-l-phenyl-butylidene)-hydrazinothiocarboxamide, 4-[(aminothioxomethyl)hydrazono]-N-(l-methylethyl)-4-phenyl-butanamide, 4-[(aminothioxomethyl)-hydrazono]-4-phenylbutanamide, 4-[(aminothioxomethyl)-hydrazono]-N-benzyl-4-phenylbutanamide, 4-[(aminothioxomethyl)-hydrazono]-N-methyl-4-phenylbutanamide, 4-[(aminothioxomethyl)-hydrazono]-N-cyclohexyl-4-phenylbutanamide, 4-[(aminothioxomethyl)hydrazono]-N,N-dimethyl-4-phenylbutanamide, l-[4-[(aminothioxomethyl)hydrazono]-l-oxo-4-phenylbutyl]piperidine, and

4-[(aminothioxomethyl)-hydrazono]-N-(l,5-dimethylhexyl)-4-phenyl- butanamide.

3. A method of treating atherosclerosis in mammals which comprises administration to a mammal having atherosclerosis a therapeutically effective amount of a compound of the formula

A wherein: R¹, R², and R³ are independently hydrogen, -C_ό alkyl, phenyl or -(CH₂)ι_-6 phenyl where phenyl is optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, - alkyl, Cj;-C₆ alkoxy, trifluoromethyl, Cι-C alkoxycarbonyl, -CO₂H or OH;

R⁴ and R⁵ are independently hydrogen, - o alkyl, C₃-C₈ cycloalkyl, -(CH₂)o-_θAr¹ where Ar¹ is phenyl, naphthyl, furanyl, pyridinyl or thenyl and Ar¹ can be optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, C,-C alkyl, phenyl, -C_ό alkoxy, phenoxy, trifluoromethyl, -C_ό alkoxycarbonyl, -CO₂H or OH, or R⁴ and R⁵ together with the nitrogen to which R⁴ and R⁵ are attached form a ring containing 4-7 carbon atoms;

and Ar is phenyl, naphthyl, furanyl, pyridinyl or thienyl which may be optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, - alkyl, C₃-C₆ cycloalkyl, phenyl, C_rC₆ alkoxy, phenoxy, trifluoromethyl, Cι-C₆ alkoxycarbonyl, -CO₂H or OH.

4. The method of treatment according to claim 3 wherein the compound used to increase HDL cholesterol is selected from:

2-(4-butylamino-4-oxo-l-phenyl-butylidene)-hydrazinothiocarboxamide, 4-[(aminothioxomethyl)hydrazono]-N-(l-methylethyl)-4-phenyl- butanamide, 4-[(aminothioxomethyl)-hydrazono]-4-phenylbutanamide, 4-[(aminothioxomethyl)-hydrazono]-N-benzyl-4-phenylbutanamide, 4- [(aminothioxomethyl)-hydrazono]-N-methyl-4-phenylbutanamide,

4-[(aminothioxomethyl)-hydrazono]-N-cyclohexyl-4-phenylbutanamide, 4-[(aminothioxomethyl)hydrazono]-N,N-dimethyl-4-phenylbutanamide, l-[4-[(aminothioxomethyl)hydrazono]-l-oxo-4-phenylbutyl]piperidine, and 4-[(aminothioxomethyl)-hydrazono]-N-(l,5-dimethylhexyl)-4-phenyl- butanamide.

5. A pharmaceutical composition for the treatment of atherosclerosis which comprises a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to the formula:

A wherein:

R¹, R², and R³ are independently hydrogen, - alkyl, phenyl or -(CH₂)_1- phenyl where phenyl is optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, Cι-C alkyl, Cj-C₆ alkoxy, trifluoromethyl, - alkoxycarbonyl, -CO₂H or OH;

R⁴ and R⁵ are independently hydrogen, Q-Q_υ alkyl, C₃-C₈ cycloalkyl, -(CH₂)o_-όAr¹ where Ar¹ is phenyl, naphthyl, furanyl, pyridinyl or thenyl and Ar¹ can be optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, Q-Q alkyl, phenyl, Q-Q alkoxy, phenoxy, trifluoromethyl, Q-Q alkoxycarbonyl, -CO₂H or OH, or R⁴ and R⁵ together with the nitrogen to which R⁴ and R⁵ are attached form a ring containing 4-7 carbon atoms;

and Ar is phenyl, naphthyl, furanyl, pyridinyl or thienyl which may be optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, Q-Q alkyl, Q-Q cycloalkyl, phenyl, Q-Q alkoxy, phenoxy, trifluoromethyl, Q-Q alkoxycarbonyl, -CO₂H or OH.

6. A pharmaceutical composition according to claim 5 wherein the compound used is selected from:

2-(4-butylamino-4-oxo-l-phenyl-butylidene)-hydrazinothiocarboxamide, 4-[(aminothioxomethyl)hydrazono]-N-( 1 -methylethyl)-4-phenyl- butanamide, 4-[(aminothioxomethyl)-hydrazono]-4-phenylbutanamide,

4-[(aminothioxomethyl)-hydrazono]-N-benzyl-4-phenylbutanamide, 4-[(aminothioxomethyl)-hydrazono]-N-methyl-4-phenylbutanamide, 4-[(aminothioxomethyl)-hydrazono]-N-cyclohexyl-4-phenylbutanamide, 4-[(aminothioxomethyl)hydrazono]-N,N-dimethyl-4-phenylbutanamide, l-[4-[(aminothioxomethyl)hydrazono]-l-oxo-4-phenylbutyl]piperidine, and

4-[(aminothioxomethyl)-hydrazono]-N-(l,5-dimethylhexyl)-4-phenyl- butanamide.

7. A process for the preparation of a compound according to the formula:

A wherein:

R¹, R², and R³ are independently hydrogen, Q-Q alkyl, phenyl or -(CH ₂) _1-6 phenyl where phenyl is optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, Q-Q alkyl, Q-Q alkoxy, trifluoromethyl, Q-Q alkoxycarbonyl, -CO₂H or OH;

R⁴ and R⁵ are independently hydrogen, Q-Qo alkyl, C₃-C₈ cycloalkyl, -(CH₂)o_-όAr¹ where Ar¹ is phenyl, naphthyl, furanyl, pyridinyl or thenyl and Ar¹ can be optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, Q-Q alkyl, phenyl, Q-Q alkoxy, phenoxy, trifluoromethyl, Q-Q alkoxycarbonyl, -CO₂H or OH, or R⁴ and R⁵ together with the nitrogen to which R⁴ and R⁵ are attached form a ring containing 4-7 carbon atoms;

and Ar is phenyl, naphthyl, furanyl, pyridinyl or thienyl which may be optionally substituted by one or more (same or different) substitutents selected from halogen, cyano, nitro, Q-Q alkyl, C₃-Q cycloalkyl, phenyl, Q-Q alkoxy, phenoxy, trifluoromethyl, Q-Q alkoxycarbonyl, -CO₂H or OH.

which comprises reacting a 4-oxo-4-arylbutyric acid of formula 1

1

wherein Ar, R^ and R^ are as defined above, with the appropriately substituted thiosemicarbazide of formula

S II H₂NNR¹^ NR²R³

wherein Rl, R^ and R- are as defined above.

8. Use of a compound of formula A, as defined in 7, as a medicament.

9. Use of a compound of formula A, as defined in 7, in the preparation of a medicament for the treatment of atherosclerosis. .