CA1080622A - Hypolipidemic compositions and method of producing hypolipidemic activity - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE

Hypolipidemic compositions containing 4-(2-thenoyl)-2,3-dichlorophenoxyacetic acid, its alkali metal salts or pharmaceutically acceptable base addition salts and methods of producing hypolipidemic activity by administering said compound. Combinations with other useful agents are also disclosed.

Description

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1 This invention re~ tes to novel hypolipidemic compo-sitions containing an active in~redient which lower plasma lipi~ concentrations and to a method of producing hypolipi-demic activity by administering nontoxic effective quantities of said ingredient to hyperlipidemic subjects. More specifi- -cally, the active ingredient used in the compositions and - methods of this invention is 4-(2-thenoyl)-2,3-dichloro-phenoxyacetic acid which has the following formula:

2CO~H

or an alkali metal salt of said acid, for example the sodium or potassium salt, or a pharmaceutically acceptable addition nontoxic salt of said acid formed with a base, for example the piperazine or (trihydroxymethyl)methylamine salt.
This acid and its preparation is described in U.S.
Patent No. 3 J 758,506. Generally, 2,3-dichloroanisole is condensed wLth thiophene-2-carboxylic acid chloride in the ~-presence of aluminum chloride, the re~ulting ketone is demethylated and ~he hydroxyketone i9 reacted with an ester of chloroacetic acid to give the product after hydrolysis of the ester. The compound iq dLsclosed as having diuretic activity.

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10~622 In accordance with the present teachings, a pharmaceutical composition is provided which comprises 4-(2-thenaly~-2, 3-dichlorophenoxyacetic acid, or a pharmaceutically acceptable salt thereof, in a dosage unit range of from about 100 mg. to 500 mg., in combination with a second active medicinal ingredient used to treat a disease in which hyperlipidemia is a significant component or with a second hypolipidemic agent, the second active medicinal ingredient is selected from the group wherein guanethidine, alpha methyldopa, phentolamine, yohimbine, phenoxybenzamine, reserpine, propranolol, hydralazine, diazoxide, tolbutamide, chlorpropamide, phenformin, allopurinol, probenacid, sulfimpyrazine, cholestyramine, nicotinamide, para-aminosalicylic acid, norethynodrel, norethindrone, norethin-drone acetate, medroxyprogesterone acetate, ethynodiol diacetate, mestranol, ethinyl estradiol, spironolactone, triamterene or chlorothalidone and a nontoxic pharmaceutical carrier.
Abnormal plasma lipid concentrations are a part of coronary heart disease and therefore the reduction of ele-vated plasma lipids is a desirable goal in the long term management of such disease. The hypolipidemic activity o~ 4-(2-thenoyl)-2,3-dichlorophenoxyacetic acid or its f - 2a -~' .

~e~8~ z 1 salts is readily demonstrated in rats by oral administra-tion of the compound at a dose of 300 mg/kg/day for 14 days.
On the morning of the fifteenth day, animals are given 150 mg/kg of compound 30 minutes prior to sacrifice. The animals are anesthetized and bled by cardiac puncture.
Livers are removed, blotted, weighed and homogenized.
Kidneys are excised, decapsulated and weighed. Plasma free - fatty acid is analyzed, and plasma and hepatic triflycerides and cholesterol are determined by standard methods.
The results of testing 4-(2-thenoyl)-2,3-dichloro-phenoxyacetic acid as described above are summarized in Table I. Compared to controls, the test compound showed a significant decrease in body weight gain, kidney weight was unaffected and liver weight was increased. Pla~ma tri-glycerides were significantly reduced but plasma cholesterol concentrations were not significantlg affected, nor was there a significant effect on free fatty acids. Hepatic - lipid concentrations are expressed as mg/g liver wet weight and~as mg/liver. The test compound did not have a signifi-cant effect~on hepatic cholesterol when expressed as mg/g liver, however total hepatic chole~terol was increased.
Hepatic triglyceride concentration was significantly reduced but total hepatic triglyceride was not affected.
In a human clinic~l study, ~-(2-thenoyl)- ~

2,3-dichlorophenoxyacetic acid has been found to decrease ~ -serum triglycexide levels at an oral daily dose of 1000 mg.
after six weeks administration.

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l~&lGtiZ;2 TABL~ 1 EFFECT OF 4~ THENOYL)-2,3-DICHLOROPHENOXYACETIC ACID ON
BODY WEIGHT GAIN, LIVER W~IGHT, KIDN~Y WEIGHT, AND PLASMA
AND HEPATIC LIPID CONCENTRATIONS
ENDPOINT CONTROL COMPOUND
0.5% Gum 150 mg/kg Tragacanth b.i.d.
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INITIAL BODY WEIGHT (g) 196 + 7 193 ~ 6 _ FINAL BODY WEIGHT (g) 286 _ 13 268 ~ 10**
- BODY WEIGHT GAIN (g) 90 + 11 74 + 6***-1 0 ~ V/ ) LIVER WEIGHT (g) 11.8 + 1.1 13.8 + 1.2**
(+17V/
.
. KIDNEY WEIGHT 2.1 + 0.2 - 2.1 + 0.1 NS
,: _ PLASMA LIPIDS
Cholesterol mg/100 ml 64 + 13 61 + 5 NS
(-5~
Triglyceride mg/100 ml 77 + 33 35 + 15**
(-54/0) Free Fatty Acid~ ~Eq/liter 338 ~ 106 287 i 116 NS
: (-33%) HEPATIC LIPIDS
Cholesterol mg/g liver 2.9 + 0.1 2.8 + 0.4 NS
( _3V/o~
Cholesterol mg/liver 33.8 _ 3.2 38.1 + 5.4*
(+13V/o~
Triglyceride mg/g liver 5.0 + 0.7 3.9 + 0.5***.
~ (-22~
Triglyceride mg/liver 58.8 + 11.0 53 8 + 9.0 NS

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NS = Not significant *p~ 0.05; **P ~ 0.01; ***P~ 0.001 compared to control group.
Values are ~ + S.D. Values in parentheses are percent change from control.
10 animals per group.

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1 The hypolipidemic compositions of this invention are prepared in conventional dosage unit forms by incorpora-ting 4-(2-thenoyl)-2,3-dichlorophenoxyacetic acid or a pharmaceutically acceptable salt thereof, in a nontoxic amount sufficient to produce hypolipidemic activity in the designated subject, with a nontoxic pharmaceutical carrier according to accepted procedures. Preferably the composi-tions will contain the active ingredient in an active but nontoxic amount selected from about 100 mg. to about 500 mg. of active ingredient per dosage unit.
The pharmaceutical carrier employed may be, for example, either a solid or liquid. Exemplary of solid carriers are lactose, terra alba, sucrose, talc, gelatin, -agar, pectin, acacia, magnesium stearate, stearic acid and the like. Exemplary of liquid carriers are syrup, peanut oil, olive oil, water and the like. Similarly the carrier or diluent include any time delay material well known to the art, such as glyceryl monostearate or glyceryl di- -stearate alone or with a wax.
A wide yariety of pharmaceutical forms can be employed. Thus, if a solid carrier is used the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form or in the form ~f a troche or lozenge. The amount of solid carrier will v2ry widely but preferably - -will be from about 25 mg. to about 1 g. If a liquid car-rier is u8ed, the preparation will be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampule or an aqueous or nonaqueous liquid suspension.

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l The method in accordance with this invention com-prises administering internally to an animal subject in need of hypolipidemic activity, i.e. a hyperlipidemic subject, the compound 4-(2-thenoyl)-2,3-dichlorophenoxyacetic acid or a salt thereof, usually combined with a pharmaceutical car-rier, in a nontoxic amount sufficient to produce hypolipi-demic activity. The àctive ingredient will be administered preferably in a dosage unit, in an active, nontoxic quanti-ty selected from about lO0 mg. to about 500 mg. of the parent chemical of Formula I. The route of administration may be orally or parenterally, the oral route being pre-ferred. Advantageou.sly equal doses will be administered two to four times daily with the daily dosage regimen being -from about 200 mg. to about 2000 mg. When the method described above is carried out hypolipidemic activity is produced with a minimum of side effects.
The pharmaceutical preparations are made following the conventional techniques of the pharmaceutical chemist involving mixing, granulating and compressing when necessary, or variously mixing and dissolving the ingredients as appropriate to the desired end product;
Also included within the scope of this invention r-are pharmaceutical compositions comprising 4-(2-thenoyl)-2,3-dichlorophenoxyacetic acid or a salt thereof as described above in combination either with a second active medicinal ingredient used to treat a disease in which hyperlipidemia is a significant component but which agent - u8ed to treat the disease or abnormal condition does not reduce the plasma lipid levels of the subject, or with a second hypo1ipidemic agene. Such disease8 and ingredients .
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~ 2 1 useful in the combination with the active ingredient of this invention are as follows:
1. Hypertension with associated hyperlipidemia a) drugs acting on the afferent sympathetic nervous system (1) ganglionic blocking agents such as guanethidine (2) centrally active drugs such as alpha . methyldopa, phentolamine or yohimbine and alpha adrenergic blocking agents such as phenoxybenzamine

(3) peripherally and centrally active drugs such as re~erpine and beta adrenergic blocking agents 8uch as propranolol : . b) drugs acting on arteriola smooth muæcle 8uch as hydralazine or diazoxide -2. Diabetes with associated hyperlipidemia . hypoglycemic agents such as tolbutamide, ` ~ chlorpropamide or phenformin ;.i 3. Hyperuricemia with associated hyperlipidemia agents such as allopurinol, probenacid or - 8ulfimpyrazine

4. Hyperlipidemia agents such as cholestyramine, nicotinamide or para-aminocalicylic acid 25. 5. Hyperlipidemia induced by oral contraceptives agents such as norethynodrel, norethindrone and acetate, medroxyprogesterone acetate, ethynodiol diacetate, mestranol or ethinyl estradiol 6. Edema treated with potas8ium sparing diuretics such as spironolactone or triamterene, or hypertension .. . .
treated with a diuretic such as chlorthalidone.

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These combination compositions will contain per dosage unit the same amount of 4-(2-thenoyl)-2,3-dichloro-phenoxyacetic acid or a salt thereof as indicated above, namely within the dosage unit range of from about 100 mg.
to 500 mg. The amount of the second active ingredient will be the same dosage set forth for the ingredient in the "Physicians' Desk Reference", 28th Edition, 1974. Specific examples of combination compositions are as follows:

4-(2-Thenoyl)-2,3-dichloro- Second Active Phenoxyacetic acid Ingredient In~ication 100 mg. to 500m.mg. allopurinol hyperuricemia 100 mg, to with hyper-300 mg. lipidemia 100 mg. to 500 mg,cholestyramine hyperlipidemia 4 g, 100 mg, to 500 mg,alpha methyldo- hypertension pa 250 mg. with hyper-lipidemia 100 mg. to 500 mg.chlorthalido- hypertension ne 50 mg. to with hyper-100 mg. lipidemia 100 mg, to 500 mg, reserpine hypertension 0.1 mg. to with hyper- -1 mg. lipidemia 100 mg, to 500 mg, triamterene edema with 25 mg, to hyp~rlipi-100 mg. demia The following examples illustrate the preparation of the hypolipidemic and combination compositions of this invention, Ingredients Mg./Table~

4-(2-Thenoyl)-2,3-dichloro 250 phenoxyacetic acid Corn starch 30 Polyvinyl pyrrolidone 12 Corn starch 16 30 Magnesium stearate 3 .

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The first two ingredients are thoroughly mixed and granulated with a 20~ w/v solution of polyvinyl pyrrolidone in water.
The wetted mass is passed through a ~4 mesh screen directly onto drying trays. The granules are dried at 50C. and mixed with the remaining corn starch and magnesium stearate, and compressed into tablets. The tablets are administered to a hyperlipidemic subject 3 times daily.
Ingredients Mg./Capsule 4-t2-Thenoyl)-2,3-dichloro- 500 phenoxyacetic acid Magnes~nm stearate 2 10 Lactose 50 The above ingredients are screened through a #40 mesh screenn, mixed and filled into #0 hard gelatin capsules.
; The capsules are administered to a hyperlipidemic subject twice daily.
Ingredients Mg./Tablet 4-(2-Thenoyl)-2,3-dichloro- 250 phenoxyacetic acid Starch 30 Povidone 12 20 Triamterene 25 Starch 16 Primojel R 12.5 Magnesium stearate 3 The ~irst two ingredients are thoroughly mixed and granulated with a 25% w/v solution of Povidone in water. The wetted mass is passed through a ~4 mesh ' _g_ .
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screen and placed onto trays. The granulated material is dried at 500C. and passed through a ~10 mesh screen.
The triamterene, Primojel, magnesium stearate and remaining starch, after passing through a #40 mesh screen, are mixed with the granulated material and compressed into tablets.

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~o~ z In accordance with the teachings of the principal disclosure, a hypolipidemic composition is disclosed which contains 4-(2-thenoyl)-2,3-dichlorophenoxyacetic acid or a pharmaceutically acceptable salt thereof and a nontoxic pharmaceutical carrier. The pharmaceutical composition may comprise in addition to the active ingredient 4-(2-thenoyl)-2, 3-dichlorophenoxyacetic acid a second active medicinal ingredient used to treat a disease in which hyperlipidemia is a significant component or with a second hypolipidemic agent together with a nontoxic pharmaceutical carrier. The preferred second active medicinal ingredient is triamterene and may be present in a dosage unit range of from 25 mg. to 100 mg. with the active ingredient the 4-(2-thenoyl)-2,3-dichlorophenoxyacetic acid in a dosage unit range of from about 100 mg. to 500 mg.
Now, and in accordance with the teachings of the supplementary disclosure, it has been found that the composition comprising 4-(2-thenoyl)-2,3-dichlorophenoxyacetic acid or a pharmaceutically acceptable salt thereof and triamterene with a nontoxic pharmaceutical carrier has been found useful as a diuretic and has further been found to have hypotensive properties. Thus, the new teachinqs now include a diuretic or hypotensive pharmaceutical composition in dosage unit form comprising per dosage unit 4-(2-thenoyl)-2,3-dichlorophenoxy-acetic acid, and an alkali metal salt of said acid or a - pharmaceutically acceptable addition salt of the acid formed with a base, in a nontoxic amount selected from about 100 mg.
to about 500 mg. and from about 25 mg. to about 100 mg. of triamterene, and a nontoxic pharmaceutical carrier.
A particularly useful diuretic combination composition of this invention contains per dosage unit 4-(2-thenoyl)-2,3-dichlorophenoxyacetic acid or a pharmaceutic-: Ai io~ z ally acceptable salt thereof in an active but nontoxic amountselected from about 100 mg. to about 500 mg. and from about 25 mg. to about 100 mg. of triamterene. This combination is also useful as a hypotensive composition. Preferably the combination composition will contain a ratio of said phenoxy-acetic acid ingredient to triamterene of 10:1. Such combination compositions have a synergistic effect on sodium excretion thereby increasing the amount of sodium excreted above the responses achieved by the single ingredients at comparable doses. The synergistic effect is demonstrated in the Na deficient rat test which measures saluretic and kaliuretic effects in rats which have been on a Na deficient diet for 4 days, fasted overnight and then given-a water load and sodium chloride providing 10 mg. Na per rat. Urine is collected for 6 hours and analyzed-for Na AND K . Results of 2 separate experiments are set forth in Table II.
In Experiment #73075, the rats on 15 and 30 mg/kg p.o.
of triamterene excreted approximately the same amount of sodium (3.31 and 3.04 mg., average pe~ group of 8). With 150~and 300 mg!kg p.o. of 4-(2-thenoyl)-2,3-dichlorophenoxyacetic acid ("phenoxyacetic acid"), the quantity of sodium excreted was less than with triamterene (0.61 and 1.71 mg.). When both compounds were combined at the low and high doses, a synergistic effect on sodium excretion was observed. After the combination of 15 mg/kg of triamterene and 150 mg/kg of phenoxyacetic acid, sodium excretion was 5.14 mg. At the higher doses, 30 mg/kg of triamterene and 300 mg/kg of phenoxyacetic acid, sodium excretion rose to 10.18 mg. This value greatly exceeds the response of the o .
- individual effects at these doses. The combination doses were also effective in lowering potassium excretion a~d elevating the ~ajK ratio. Wlth either triamterene or phenoxyacetic acid ~ 12 .

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alone the Na/K ratio did not rise above 1.93. After low and high doses of the combination these values were elevated to 3.41 and 9.03 respectively.
In Experiment $92475, 15 and 20 mg/kg p.o. of triamterene produced a dose response effect on sodium excretion (2.91 and 4.88 mg.). The responses to 150 and 300 mg/kg p.o. of phenoxyacetic acid were 0.78 and 1.18 mg. The combination of 15 mg/kg of triamterene and 150 mg/kg of phenoxyacetic acid resulted in a sodium excretion of 5.79 mg. At the higher doses (30 mg/kg of triamterene and 300~mg/kg of phenoxyacetic acid) of the combination the output of sodium was 7.47 mg. Again, the combined effect was much more pronounced than the natriuretic -response seen with each compound alone. Potassium excretion was also less with the combination.
With either compound alone the highest Na/K ratios were observed with triamterene (2.32 and 3.48). Values for the low and high doses of the combination rose to 5.32 and 6.22.

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TABLE II
Sodium Deficient Rat Test 4-(2-Thenoyl)~2,3-Dichlorophenoxyacetic acid + Triamterene Sodium Potassium Dose Excreted Excreted Na/K
Drug mg/kg p.o. (m~)* (mg)* Ratio Experiment #73075 Control ---_ 0.13 3.99 0.6 Triamterene 15 3.31** 2.88 1.93 3.04** 4.18 1.23 Phenoxyacetic150 0.61 3.07 0.34 acid 300 1.71 8.88 0.32 Triamterene15 ) 5.14** 2.59 3.41 +
Phenoxyacetic150 ) acid Triamterene30 ) 10.18** 1.91 9.03 +
Phenoxyacetic300 ) acid Experiment #92475 Control --- 0.25 2.37 0.18 Triamterene 15 2.91** 2.13 2.32 4.88** 2.35 3.48**
Phenoxyacetic150 0.78 2.17 0.64 acid 300 1.18 5.17 0.39 Triamterene15 ) 5.79** 1.82 5.32**
Phenoxyacetic150 ) acid Triamterene30 ) 7.47** 2.06 6.22**
Phenoxyacetic300 ) acid .~ I
* Average per group ** Significant p= < 0.05 8 ar,imal~ per group ... 1~ ~

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~OB(J~Z2 The invention has been described in detail with particular reference to preferred embodiments thereof, but it will be understood that variations and modifications can be effected within the spirit and scope of the invention.

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Claims (4)

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A pharmaceutical composition comprising 4-(2-thenoyl)-2, 3-dichlorophenoxyacetic acid, or a pharmaceutically acceptable salt thereof, in a dosage unit range of from about 100 mg. to 500 mg., in combination with a second active medicinal ingredient used to treat a disease in which hyperlipidemia is a significant component or with a second hypolipidemic agent, said second active medicinal ingredient being selected from the group is guanethidine, alpha methyldopa, phentolamine, yohimbine, phenoxybenzamine, reserpine, propranolol, hydralazine, diazoxide, tolbutamide, chlorpropamide, phenformin, allopurinol, probenacid, sulfimpyrazine, cholestyramine, nicotinamide, para-aminosalicylic acid, norethynodrel, norethindrone, norethindrone acetate, medroxyprogesterone acetate, ethynodiol diacetate, mestranol, ethinyl estradiol, spironolactone, triamterene or chlorothalidone, and a nontoxic pharmaceutical carrier.

2. A pharmaceutical composition according to claim 1 in which the second active medicinal ingredient is triamterene.

3. A pharmaceutical composition according to claim 2 in which the triamterene is present in a dosage unit range of from about 25 mg. to 100 mg.

4. A pharmaceutical composition according to claim 3 comprising 250 mg. of 4-(2-thenoyl)-2,3-di-chlorophenoxyacetic acid and 25 mg. of triamterene per dosage unit.