GB2208473A - Anti-hyperlipemic agent - Google Patents

Description

V 4 i 1 ANTI-HYPERLIPEMIC AGENT r_ 11. I- 22 U z, 4 / j The present

invention relates to the use of dehydroepiandrosterone (DHA) as an anti-hyperlipemic agent, and more particularly relates to the use of DHA as an anti -hyperl ipemic agent to be used for the treatment and prophylaxis of hyperlipemia in human or animal cases.

Normal ranges of serum lipid content and lipoprotein content in humans are shown in Table 1 below:

TABLE 1 - Normal Ranqes of Serum Lipid Content and Liponrotein-Content in Human Kind Total cholesterol Triglyceride Phospholipid Chylomicron VLDL (VLDL Chol.) LDL (LDL Chol.) HDL Male (HDL - Chol.) Female (HDL Chol.) Lipoprotein Fraction alpha-lipoprotein (HDL) pre beta-lipoprotein (VLDL) beta-lipoprotein (LDL) Normal Range - 211 mg/dl - 160 mg/dl 159 299 mg/dl 0 (at hungry) - 400 mg/dl less than 30 mg/dl 200 - 400 ng/dl less than 170 mg/dl - 425 mg/dl 37 - 57 mg/dl 250 - 650 mg/dl 36 mg/dl 20.0 - 50.0 % 8.5 - 19.9 % 37.1 - 54.7 % (Tokyo Medical and Dental University) Physiocochemical properties and details of the compositions of these lipoproteins are as shown in Table 2 below:

Table 2 Physicochemical Properties and Composition of Lipoprotein Lipoprotein Chylomicrom VUL IDL LDL HDL? HM, Specific gravity (g/mi) <0.96 0.96-1.006 1.006-1.019 1.019-1.063 1.063-1. 125 1.125-1.21 Diamiter () 800-10000 300-750 ' 220-300 190-220 85-100 75-85 Electrophoresis origin pre,4 (mid band) Composition (%) Trigryceride M) 85 55 24 10 5 4 Cholesterol Estel (CE) 5 12 33 37 18 12 Free Cholesterol (M 2 7 13 a 6 3 Mcapholipid M). 6 1.8 12 22 29 23 Protein 2 8 18 23 42 58 VUL: Very low density lipoprocein IDL: Intermediate density lipoprocein LDL: Low density lipoprotein HM High density lipoprotein 4 m 3 Hyperlipemia. is a condition in which a surplus quantity of lipids is present in blood due to excessive uptake of lipids such as cholesterol, neutral fats, etc or due to abnormal lipometabolism. Hyperlipemia is a generic name for all of these conditions.

Hyperlipemia. is further divided into hyperchylomicronemia, hypercholesteremia, hypertriglyceridemia, etc according to the type and concentration of the lipid(s) in excess. The WHO (World Health Organisation) classifies hyperlipemia into 6 types. as shown in Table 3 below, according to the condition of lipoprotein.

Table 3 Classification and Features of Hyperlipemia Lipid concentration (Classification by WHO) Image Filter Trigly- Causes Type of paper Condition of CITG lipo- electro- lipids terol ceride proteins phoresis (220mg/d1) (120mgIdl) Increase of Hyperchylomicronemia or-'P- <0.2 Decrease of ability to dispose chyLomicron chyLomicron Decrease of lipase activity A-lipoprocein lIa Increase of)S- Hypercholesceremia TTT ort catabolic disorder LDL Increase of 8 Hypercholesteremia TG synthesis enhancement LDL and pre Hypertriglyceridemia and derease of catabolism Ilb VLDL Decrease of lipoprotein catabolism rncresse of Broad H"ercholesteremia 1- Z Existence of abnarm, III IDL Hypertriglyceridemia lipoprotein pre-,6 catabolic disorder Increase of preS Hypertriglyceridemia e->or TG catabolic disorder Partial enhancement of IV VLDL ft synthesis Increase of Ryperchylomicronemia Decrease of lipase activity v chylomicron Hypercholesteremia 0.15<0.6 TG catabolic disorder and VUL Hypertriglyceridemia In general, Ila > IV > Ilb h.

k' 1 1 1 1 Hyperlipemia is generally observed in people of middle age and older people, in most cases; lipids are deposited on arterial walls or connective tissues. on the other hand, hyperlipemia is seen in younger people and is mostly a metabolic disorder of a hereditary nature: it is called familial hyperlipemia. Hyperlipemia is closely related with arterial sclerosis and very often causes pultaceous sclerosis in aortae, coronary arteries, basilar arteries, etc. it frequently occurs in the coronary artery and causes angina pectoris and myocardial infarction. Further, it also tends to cause cholelithiasis.

As already described, hyperlipemia causes vascular disorders such as arterial sclerosis. Accordingly, it is necessary or desirable to take therapeutic measures to cope with each of the causes.

To this end, therapeutic methods can be conveniently divided as follows:

(1) In order to treat dietary hyperlipemia, which is caused by excessive uptake of lipids or cholesterol, alimentary therapy (restriction of food intake) or kinesitherapy (increasing energy consumption) is recommended. If neither of these therapeutic methods do not bring any favourable results, pharmacotherapy is recommended.

(2) To treat the hereditary familial hyperlipemia, which involves a disorder in a lipometabolic enzyme, or apoprotein or a defect in the receptor mechanism, pharmacotherapy by direct 6 administration of drugs is performed.

(3) To treat secondary hyperlipemia, which is caused by the other diseases such as nephrosis, diabetes, etc, therapy to eliminate the causes of the primary disease should be undergone to improve the pathological conditions of the primary disease.

Drugs useful in pharmacotherapy include drugs to inhibit intracorporeal absorption of cholesterol, drugs to inhibit biosynthesis of cholesterol and drugs to promote lipometabolism. Absorption inhibitors include cholestyramine made of anion-exchange resin, melinamide derived from linoleic amide, and soysterol, which is an unsaponifiable substance of soybean oil. Biosynthesis inhibitors include probucol and clofibrate preparations, which are widely known. Lipometabolism promoters include heparin and dextran- sulphur ic acid, which increase the activity of lipoprotein lipase and enhance.the catabolic effects of lipoprotein.

However, it is also known that these drugs can cause side effects such as mild symptoms in the digestive tract (in the case of cholesterol absorption inhibitors), liver disorders, digestive tract disorders and cholelithiasis (in the case of cholesterol biosynthesis inhibitor).

The present inventors have carried out biochemical research on hyperlipemia and have found that DHA and DHA-sulphate (DHA-S) have excellent anti -hyperl ipemic effects. Further, the inventors have developed DHA preparations with which to perform therapeutic t 7 experiments on human subjects and have evaluated the effects and side effects by administering the drugs to patients with hyperlipemia. The results of the experiments revealed that DHA preparations have the effect of relieving hyperlipemia, strengthening the vascular wall and removing thrombocytes attached to the vascular wall. In addition, no unacceptably adverse side effects were observed.

For a better understanding of the present invention, and to show how it may be put into ef f ect, ref erence will now be made by way of example to the accompanying drawings, in which:

Figure 1 shows a synthetic for steroids in the adrenal cortex, and Figure 2 shows the metabolic mechanism of dehydroepiandrosterone.

The present invention relates to the anti-hyperlipemic use of DHA or a derivative thereof. The term "derivative" as applied to DHA in this specification includes DHA conjugated with sulphuric acid and similar products.

DRA as mentioned herein is a chemical substance already known. It is one of the steroid hormones produced and secreted by the adrenal cortex. Derivatives of DHA include esters on the third hydroxyl radical of DHA. A typical example is DHA conjugated with sulphuric acid which is a urophanic type of DHA. However, derivatives are not limited to this; synthesised derivatives may be 1 4 8 used.

In the following description, which is given by way of example, properties of DHA and DHA-sodium sulphate, which are two effective components according to the present invention, will be described.

(1) Dehydroepiandrosterone (DRA) Chemical name Molecular formula Molecular weight Structural formula 3 beta-hydroxy-5-androsten-17-one C19H2802 288.43 0 HO "c Melting point Needle crystals Leaflet crystals Property 1410C 152 1530C White crystals or crystallin powder with no odour Easily soluble in benzene, alcoholand ether. Insoluble in water.

Solubility (2) Dehydroepiandrosterone-sodium sulphate (DRA-S) Chemical name 3 beta-hydroxy-5-androsten17-one-sodium sulphate dihydrate C19H27NaOSS. 2H20 Molecular formula:

9 Molecular weight: 426.50 Structural formula:

NoO.3SO Melting point Property Solubility pH -0 "d - 2H,20 Approx. 1540C (decomposed) White crystals or crystalline powder with no odour and bitter taste. Slightly soluble in methanol, not very soluble in water, hardly soluble in dehydrated ethanol, and substantially insoluble in acetone, ether, chloroform or benzene. 4.0 - 7.0 (0.5 W/V% aqueous solution) Normal dosage of DHA or a derivative thereof based on this invention would be from 25 to 75 mg/day in oral administration. The dosage may be adjusted according to sex, age, degree of obesity, and other symptoms.

Oral administration is generally suitable. For oral administration, the product may be prepared in tablets, capsules, granules, powder, etc. The preparations can be produced by conventional pharmaceutical procedures.

An approximate synthetic pathway for adrenal cortex steroids is shown in Figure 1. About 50 different steroid hormones biosynthesized in the adrenal cortex have been isolated up to now. As shown in Table 4, they are roughly divided into glucocorticoids (for example, cortisol), which are biochemically related to saccharometabolism, mineral ocorticoids (eg aldosterone), which are related to electrolyte metabolism, and androgens, which would include DHA and DHA conjugated with sulphuric acid (DHA-S).

TABLE 4 - Steroid Secreted from Human Adrenal Cortex mean amount secGroup Cznpound reted by adult mr dav cortisol 15 - 20 mg glucocorticoid corticosterone 2 - 5 mg aldosterone 50 - 150 ug mineralocorticoid 11-deoxycorticosterone trace dehydroepiandrosterone 15 - 20 mg androgens delta4-androstenedione 0 - 10 mg 11 beta-oH androstenedione 0 - 10 mg pregnenolone 0.5 - 0.8 ng progrestin 17-OH pregnenolone 0.2 - 0.4 mg estrogen estradiol trace DHA can be produced from DHA-S.

For this reason, DHA-S A is called a depot or storage form of DHA. DHA i s a steroid which can be a biosynthetic origin of the androgenic hormones testosterone and estradiol, but testosterone and estradiol are synthesised in the adrenal cortex in small quantities only.

DHA has very few effects as a sex hormone. It has effects on lipometabolism and protein metabolism and also exerts an influence on the metabolism of salts such as phosphates, potassium salts and sodium salts. It is excreted into urine as DHA-S, as shwon in Fig. 2. It is also excreted in the form of androsterone or etiocholanolone conjugated with sulphuric acid or glucuronic acid through androstenedione. All of these substances can be determined as 17-KS (ketosteroids). In case of women (not pregnant), almost all of these 17-KSts are said to be derived from the adrenal cortex. In case of men, 2/3 to 3/4 of them are derived from the adrenal cortex, whereas the remaining 1/3 to 1/4 come from the testes.

It has been experimentally demonstrated that only a low concentration of metabolic products of DHA are excreted into urine (17-KS in urine). When diabetes or other diseases are combined with obesity, it is found in extremely low concentrations.

DHA and DHA-S (ie, the storage type of DHA) exist abundantly in the blood of younger people (20 to 30 years old), but the values gradually decrease as they get older. In contrast, cortisol is secreted at an almost constant level (15 - 20 mg per day) irrespective of age, and 17-OHCS (hydroxycorticoid) is excreted into 12 urine as its metabolite.

DHA inhibits synthesis of lipids and reduces cholesterol and lipids in blood whereas excessive secretion of cortisol, coupled with excessive secretion of insulin against it, enhances snythesis of lipids. Accordingly, when DHA secretion decreases due to age or other reasons, cortisol and insulin cannot be controlled or inhibited any more and lipids begin to accumulate gradually in fat tissues. This results in obesity, causes troubles in production and secretion of insulin, and finally induces diabetes.

DHA has the effect of inhibiting and controling production of insulin and cortisol at the same time. In addition, it hinders blood coagulation and platelet aggregation.

When given orally, DHA reaches the intestine and is turned,to DHA-S there. It is then transported to various tissues in this form and is reconverted to DHA again. This inhibits the activity of the enzyme glucose-6-phosphate dehydrogenase (G6PDH), which is closely implicated with steatogenesis. This is bel.ieved to give DHA its anti -hyperi ipenic properties. Another useful feature of DHA is that no side effects were observed during this experiment.

The anti-hyperlipemic effects of DHA appear not only in human cases but also in animals kept at home or in the zoo, so it is useful as an antihyperlipemic agent both for human and veterinary cases.

z Z f 13 In the following, production examples of the antihyperlipemic agent according to the present invention, animal experiments, acute toxicity tests and clinical tests are described. It should be understood that this invention is not limited to the exact details of such description.

Production Example - DHA tablet Composition of a tablet: DHA (dehydroepiandrosterone) Lactose Starch Polyvinyl pyrrolidone-K30 Maqnesium stearate Total mg 80 mg 12. 5 mg 5 mg 2. 5 mcf 125 mg Tablet production method Tablet diameter Appearance Production Example 2 - DHA-S tablet Composition of a tablet: DHA-S (dehydroepiandrosteronesodium sulphate) Wet granule method 7 mm White tablet mg 70 mg 12.5 mg Lactose Starch Polyvinyl pyrrolidone-K30 5 mg Maqnesium stearate 2.5 mg Total 125 mg Tablet production method: Wet granule method Tablet diameter: 7 mm k 14 Appearance: White tablet 35 mg of DHA-S is equivalent to 25 mg of DHA.

Animal Test Twenty adult SD rats (10 male rats and 10 female rats) were used, being divided into a control group and a test group. The feed containing DHA (0. 6%) was given to thetest group and the feed without DHA was given to the control group for 11 weeks. Then, the rats were examined.

The composition of rat body in DHA-administered group is as given in Table 5.

TABLE 5

Sex male (rr-5) female (rF-5) group control DHA- control DHA qr-OUP adm. = stered C1roulD administered gro C1rouD body weight (g) 526 445 288 268 water 58.3 62.2 60.3 64.6 protein 51.5 56.2 49.3 59.0 fat (%) 36.8 33.2 37.4 24.0 An experiment was performed on the effects of DHA to determine liver weight, G6PM activity in liver tissue and triglycerides in serum. The results were as shown in Table 6.

1 1 1 is TABLE 6 sex male fe 9 control DHA- control DHA CWOUP administered CWOUP administered cf-roup CWOUP liver weight (g) 0.45 0.77 0.37 0.51 G6PM activity 0.0486 0.0301 0.0473 0.0303 (=m/ing protein) triglycerides 215 290 190 in serum (mcj/dl) Acute Toxicitv Test DHA was given subcutaneously or orally to male and female ICR mice and SD rats and the LD50 (mg/kg) was determined. The results are given in Table 7.

TABLE 7 mouse rat route male femle nale fe subcutaneous 900 1,060 1,000 1,015 administration Oral administration >10,000 >10,000 >10,000 >10,000 16 In the chronic toxicity test with mice and rats, no pathological change was observed in any cases.

Clinical Test 1 A clinical test was performed on 5 patients with hyperlipemia. The age, sex, amount of lipid in blood, etc are shown in Table 8 below. The drug used for the test was DHA tablets prepared according to Production Example I above, each containing 25 mg of DHA. One tablet was given at every meal, 3 times per day (75 mg of DHA given per day). The administration period varied from 3 to 5 months according to the symptoms. The use of the other drugs was avoided during this period.

1 f 17 Table 8 test number 1 2 3 4 5 age 64 52 56 48 44 sex man woman man woman man total cholesterol (mgldl) 290 250 280 260 300 before triglyceride (neutral administration fats) (mgIdl) 257 270 260 260 280 free fatty acid (1A1Eq/1) 720 590 680 580 690 total cholesterol (mgldl) 205 230 190 210 210 after triglyceride (neutral administraion fats) (mgldl) iso 130 140 120 150 free fatty acid (/t( Eq/1 510 430 490 450 53 j 0 Clinical Test 2 Clinical test was further performed on 5 woman patients. The drugs used for the test was said tablets. One tablet was given one time per day (25 mg of DIIA given per day). Table 9 below shows the test results of improvement of lipids in blood. As in the case with the clinical test 1 above, use of the other drugs was avoided during the test period.

Table 9

Test No.

Sex Age.

Administration period (days) Free 9-lipo- Trigly- Phospho- fatty acid E-type HM Division protein ceride (M) lipid (PL) (NEFA) chol. chol.

(mgldl) (mg/dl) (mgldl) (/X Eqlml) (mgIdl) (mgIdl) 6 woman 24 14 Before 543 229 213 0.798 148 33 administration After 461 147 189 0.460 141 38 administration 7 Before. 322 127 221 0.610 116 44 administration After 358 73 200 0.942 140 54 administration Before 494 86 261 0.110 206 53 administration a, 4a 58 After 469 98 198 0.319 155 43 administration 9 woman 45 76 woman 50 78 1.

Before 255 122 170 0.459 ill 4 administration Af ter 283 106 217 0.507 133 58 administration Before 596 197 237 0.136 179 51 administration Afr-!r 455 154 220 0.499 190 53 administration 1 19 In each case, the values of triglyceride, phospholipid and the like which have been beyond the normal ranges are lowered to be within normal ranges and it was confirmed that DHA has an excellent anti-hyperlipemic effect.

As described above, when the anti-hyperlipemic agent based on the present invention is given to patients, its efective component DHA or its derivative improves lipid metabolism and normalizes lipid level in blood. As a result, the drug reinforce blood-vessel walls to prevent arterial sclerosis as well as angina pectoris and myocardial infarction in human and animal cases.

1 1 1 h 9

Claims (9)

1. The use of dehydroepiandrosterone or a derivative thereof in the preparation of an anti-hyperlipemic agent.

2. The use as claimed in Claim 1 wherein the derivative is an ester on the third hydroxyl radical of DHA.

3. The use as claimed in Claim 2 wherein the derivative is dehydroepiandrosterone conjugated with sulphuric acid.

4. The use as claimed in Claims 1, 2 or 3, where the anti-hyperlipemic agent is for human use.

5. The use as claimed in Claim 1, 2 or 3, wherein the anti-hyperlipemic agent is for veterinary use.

6. The use as claimed in claim 1, wherein dosage of the dehydroepiandrosterone or the derivative is 25 - 75 mg/day/human.

7. The use as claimed in any one of claims 1 to 6, wherein the antihyperlipemic agent is orally administerable.

8. The use as claine din Claim 7 wheein the agent is in the form of tablets, capsules, granules or powder.

9. The use of dehydroepiandrosterone or a derivative thereof substantially as herein described with reference to the examples.

Published 1988 at nIte Patent Office. State House 66 -- H.:-. M'CIR 4TF Further c,-rcc rna--., be &,tai-neJ- frcin The Patent Office Sales Brancli, St Marv Cray. Orping-cit. Kent BR5 3RD- Printed by Mtiltilplex techniques ltd', St Mary erky. Kent Con- 1877