DE3410850A1 - MEDICINAL PRODUCTS FOR REINFORCING AND USING BLOOD VESSELS - Google Patents

Description

HOFFMANN · EITLE & PARTNER

PATENT AND LAWYERS PATENTANWÄLTE DIPL.-ING. W. EITLE DR. RER. NAT. K. HOFFMANN DIPL.-ING. W. LEHN DIPL.-ING. K. FOCHSLE DR. RER. NAT. B. HANSEN · DR. RER. NAT. H -A. BRAUNS DIPL.-ING. K. GORG DIPL.-ING. K. KOHLMANN - LAWYER A. NETTE

_3_ 40 014 o / sm

Otsuka Pharmaceutical Co., Ltd. Tokyo / Japan

Medicines for strengthening blood vessels and their use

The invention relates to a pharmaceutical composition for Strengthening the blood vessels, the active ingredient at least one fatty acid or a salt thereof of the general Formula 1)

H- (CH _n ). (CH-) -COOH

2 m \ S ^ c η

wherein the carbon-carbon bond of the formula

a single bond or double bond (cis form), m denotes 0 or 6 and η 7 or 8 and pharmaceutically acceptable Includes carrier.

The pharmaceutical compositions of the invention are useful prophylactic agents and remedies for treating hypertonic vascular injuries such as cerebral apoplexy Etc.

ARABELLASTRASSE A · D-BOOO MDNCHEN 81 · TELEPHONE CO8BJ 011087 · TELEX Ö-2961O CPATHE> ■ TELECOPIER 0183

Cerebral apoplexy is a serious hypertensive disease and a leading cause of death in Japanese Population. Cerebral apoplexy results in high mortality when it occurs, and even when it does not fatal, the disease is difficult to cure completely and patients are often bedridden stay and rely on the help of other people. Therefore, cerebral apoplexy is one of the incurable diseases that occur in a high percentage of the population. So far there is no sure way to cure this condition other than to seek preventive treatment before it occurs undertakes.

The development of high pressure is due to the mutual influence of genetic and environmental factors caused. Genetic factors that occur in hypertension and cerebral apoplexy have been implicated in stroke prone spontaneous hypertensive rats (hereinafter referred to as SHRSP) were examined. (100% of these rats develop severe hypertension spontaneously and over 90% of them develop cerebral apoplexy and die from it: Japanese Circulation Journal, Volume 41, Pages 259-266, March (1977)). As a result, takes one assumes that the genetic · factors involved in hypertension occur and those that are present in cerebral apoplexy are present differently from one another, and that these two kinds of genetic factors are closely related. When a person both If this has genetic factors, then that person is more prone to developing cerebral apoplexy. If, on the other hand, the extent to which the

genetic factors contributing to high blood pressure is not that great (i.e., in other words, if the person not suffering from severe hypertension) then the person will in the event that the contribution of genetic Factors for which cerebral apoplexy is high, cerebral apoplexy afflicted even if not under severe High pressure suffers.

Medicines for the treatment of hypertensive diseases

1.0 gen "currently in use are mainly Extreme pressure agent. From the results of the aforementioned studies with SHRSP for combating high-

. pressure disorders, it turns out that the use of hypertensive drugs is insufficient, and that drugs with a completely different mode of action are additionally required. As one Medicines can be viewed as blood vessel-strengthening agents, i. H. a medicine that is used for treatment used to treat high blood pressure, but does not prevent vascular brain damage from occurring. In the high pressure stage, a significantly greater load is placed on the blood vessel system (in particular on the arterial system) than normal, and functional dyscrasia of the cell membrane appears, such as hyperpermeability of the blood vessel wall cells, and as a result, blood vessel necrosis takes place instead, through which the blood vessels rupture and bleeding (cerebral haemorrhage) occurs and / or thrombus formation (cerebral infarction) in the necrotic areas of the blood vessels or in others Place. To treat these symptoms and syndromes,

- D ~ "

Considered the blood vessel-thickening agent.

The currently known blood vessel-strengthening agents are known compounds of the vitamin P group such as rutin, hesperidin, eriocitrin and the like However, ß-enhancing agents are bad in the bowels absorbed and rapidly excreted when administered by injection unfortunately its effect as a blood vessel-strengthening agent

10 to be regarded as low. In addition, these blood vessel strengthening agents cause side effects such as anaphylaxis for blood vessel strengthening agents,
Digestive difficulties and the like, so that they are not satisfactory. Therefore, from the present

carried out thorough investigations on the inventors, to find new blood vessel-strengthening agents and as a result, the compounds became general Formula (1) found to have prophylactic activity against blood vessel damage in hypertensive diseases and which can be used as effective blood vessel-strengthening agents. On this knowledge the present invention is based.

The object of the invention is to provide pharmaceutical compositions for blood vessel-strengthening agents which contain as active ingredient at least one fatty acid or a salt thereof of the general formula (1) in addition to a pharmaceutically acceptable carrier.
30th

Another object of the present invention is to

to provide a pharmaceutical composition, which are suitable for the treatment of hypertonic vascular diseases such as cerebral apoplexy.

These and other objects of the invention are set forth below Description and explained in more detail in the figures.

In the figures: 10 '

Fig. 1 shows a graph showing the relationship between

the blood pressure and the age (days) of SHRSPs fed with Diet-I and Diet-II, respectively.
15th

Fig. 2 shows curves showing the relationship between blood pressure and age (days) of SHRSP, the were fed a Diet-I and were chronically administered methionine. 20th

Fig. 3 shows graphs showing the relationship between the rate of occurrence of cerebral apoplexy and the survival time of SHRSP to which Diet-I and Diet-II or Diet-III were administered, indicates.

Fig. 4a shows curves showing the relationship between the Blood pressure and the age (days) of SHRSP to which Diet-I or Diet-IV were administered is indicating.

Fig. 4b is a graphic representation and shows the Relationship between the rate of occurrence of cerebral apoplexy from SHRSP and the type of Diets (Diet I and Diet IV). 5

Fig. 5 shows curves showing the survival rate of SHRSP, who were fed a diet containing palmitoic acid.

Fig. 6 shows curves for the survival rate of SHRSP obtained with a diet containing undecylenic acid or palmitin drinks had been fed.

The present invention relates to medicaments for strengthening blood vessels, which are used as the active ingredient at least one fatty acid or a salt thereof in general Formula 1)

20 s

wherein the carbon-carbon bond of the formula

is a single or double bond (cis form), m is O or 6 and η is 7 or 8 and contains a pharmaceutically acceptable carrier.
.

The fatty acids or their salts of the general formula

(1) include:

cis-9-hexadecenoic acid {palmitoleic acid, in which the .Carbon-carbon bond of the formula

■> <

is a double bond, m is 6 and η 7), n-hexadecanoic acid (palmitic acid in which the carbon Carbon bond of the formula

a single bond, m 6 and η 7) and 10-Undec; en acid (undecylenic acid, in which the carbon Carbon bond of the formula

15 ^

a double bond, m is O and η 8).

The drug composition of the present invention for strengthening blood vessels causes recovery of the Fragility and the abnormal permeability of the blood vessels, prevents the aging of the blood vessels and results excellent results in the prophylaxis and therapeutic treatment of various blood vessel diseases in humans and animals, e.g. B. in bleeding caused by the fragility of the blood vessels will.

The compositions according to the invention are also . 30 for strengthening blood vessels are particularly effective for damage to cerebral vessels, and they are suitable prophylactic agents and therapeutics for cerebral apoplexy.

The aforementioned active ones. Components, d. H. the fatty acids or their salts of the general formula (1) have have improved visceral absorbency and are stable in the blood and therefore a very strong good duration of the pharmacological effects and a low toxicity, so that they can be used for a longer period of time Can occupy periods. Some of the fatty acid salts of general formula (1) are in rather large ones Amounts in a number of green plant and animal oils and fats, typically milk fats, fish oils, Whale oil, apples and the like. Contain and can from these raw materials in the usual way, for. B. by molecular distillation, Countercurrent distribution, chromatography and the like. Can be obtained and are used as standard reagents available in the stores. For practical purposes it is not necessary to use the fatty acids or their Salts, which are used as active ingredients here, to isolate or purify, rather they can be called Raw products that contain small amounts of other fatty acids

20 ren etc. included, use.

The fatty acids or their salts of the general formula (1) can also contain synthetic organic products made from suitable starting materials.

Pharmaceutically acceptable salts of the fatty acids of general formula (1), which are used as active ingredients in the Medicaments according to the invention for strengthening the blood vessels are typically such pharmaceutically acceptable salts as alkali salts, e.g. Sodium salts and potassium salts, alkaline earth salts, such as kai

zium salts and other metal salts such as aluminum salts, ammonium salts, amine salts such as morpholine salts, piperadine salts, Trimethylamine salts, diethylamine salts and the like.

The fatty acids or salts of the general formula (1) according to the invention can be administered alone or with any suitable carrier or diluent and they can also be administered as pharmaceutical compositions consisting of a mixture of the Fatty acids or their salts represented by the general formula (1) as the active ingredient besides pharmaceutically acceptable ones Carriers are present. Examples of pharmaceutically acceptable carriers are diluents and excipients such as fillers, volume-increasing agents, binders, humectants, disintegrants, surface-active agents Agents, lubricants and the like. These substances are all used as additives in the manufacture of Medicines are known in the desired administration forms. The administration forms of the invention Medicines can vary and be adapted to the respective application. Typical examples the forms of administration are tablets, pills, powders, granules, capsules, suppositories and others Solid and / or semi-solid preparations;

25 liquids, suspensions, emulsions, injections

(Solutions, suspensions) and the like. Administration in the form of a solid is particularly preferred. When shaping, excipients such as lactose, refined sugar, sodium chloride, a glucose solution , urea, starch, potassium carbonate, kaolin, crystalline cellulose, can be used as carriers,

Silicic acid etc., binders such as water, ethanol, propanol, simple syrup, a glucose solution, glycol, Glycerine, a starch solution, a gelatin solution, carboxymethyl cellulose, Shellak, methyl cellulose, calcium phosphate, polyvinylpyrrolidone etc. Suitable disintegrants are starch, sodium alginate, agar powder, laminar powder, Not-triuminous bicarbonate, potassium carbonate, fatty acid esters of polyoxyethylene sorbitol, sodium lauryl sulfate, Monoglyceride of stearic acid, lactose etc. Disintegration inhibitors are, for example, refined sugar, Stearin, cocoa butter, hydrogenated oils, etc. Absorption accelerators are, for example, quaternary ammonium bases, Sodium lauryl sulfonate etc. humectants are, for example, glycerine, starch etc. absorbents are, for example, starch, lactose, kaolin, bentonite, colloidal silica etc. and suitable Lubricants are, for example, purified talc, stearic acid salt, boric acid powder, macrogels, solid Polyethylene glycol etc. If necessary, the tablets can be provided with a conventional coating and as coated tablets, e.g. As sugar-coated tablets, gelatin film-coated Tablets, as coated with an enteric layer, ne tablets, as film-coated tablets, as double-layer tablets, as multi-layer tablets etc.

When shaping into pills, carriers, for example excipients such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oils, kaolin, talc, etc., and binders such as powdered gum arabic, powdered Tragacanth, gelatine, ethanol etc. can be used and laminariums,

Agar etc. can be used. In the production of suppositories, for example, polyethylene glycol, Cocoa butter or higher alcohols or esters of higher alcohols, gelatin or semi-synthetic Glycerides used. When making injections, one can make solutions or suspensions, which are then preferably sterilized and adjusted to be isotonic with blood.

In the preparation of injections for solutions, emulsions and suspensions one can use as a diluent for example water, ethyl alcohol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, Polyethylene sorbitol fatty acid ester etc.

use. You can add sodium chloride, glucose or glycerin to the injectable solutions in a sufficient amount Add amount to make it isotonic to blood. In the production of pastes, creams or Gels can be used as diluents, for example, petroleum jelly, paraffin, glycerine, cellulose derivatives, polyethylene glycol, Use silicone, bentonite, etc. If necessary, the medicaments according to the invention for strengthening the blood vessels can also contain antioxidants, z. B. butylated hydroxytoluene, propyl gallate, quinone, a-tocopherol etc., common dissolving agents, buffer solutions, analgesics, preservatives, coating agents, fragrances, flavorings, sweeteners and other drugs, e.g. B. Platelet Aggregation Inhibiting Funds etc. included.

The amount of active ingredient of the general formula

mel (1) in the medicaments according to the invention is not particularly limited and can be selected within a wide range. In general, 0.01% by weight or more of the active compound calculated as the free fatty acid is used. As tablets contain the tablets for example about 0.01 to 1.0 g each of active compound expressed as free fatty acid.

The administration of the medicaments for enhancing blood vessels of the present invention is not particularly limited and one can employ the appropriate methods for the respective administration forms. Tablets, pills, solutions, suspensions, emulsions, granules and capsules can be used administered orally; Injections are administered intravenously, individually or together with usual ones injectable transfusions such as glucose solutions, amino acid solutions etc. and if necessary the injections are intramuscular, intracutaneous, subcutaneous or administered intraperitoneally. Suppositories are administered rectally and, if necessary, vaginally.

The dosage of the medicament according to the invention depends on the condition of the patient etc. In general, the active ingredient, expressed as the free acid, is in an amount of about 1 to 200 mg per kg of body weight per day and preferably in amounts of 4 to 150 mg per kg of body weight administered per day. In general, the aforementioned daily amount of active Components divided into 3 to 4 daily doses.

The fatty acids or their salts of the general formula (1) which are used as active ingredients in the

moderate compositions for strengthening blood vessels can be determined on the basis of the results obtained from a number of pharmacological studies .using SHRSP in connection with the blood Evaluate pressure., The index of cerebral apoplexy and other facts caused by the activities, where a number of test diets are administered, each with a different composition of the Fatty acids or their salts contain an active ingredient.

The blood vessel-strengthening effects of fatty acids or of the salts according to the general formula (1), which are used as active ingredients in the medicaments according to the invention can also be contained by the physical and morphological. Properties to be assessed, e.g. B. by the tear strength of a sample of a SHRSP that has received the respective trial diets Aorta.

The invention is based on the following pharmacological Studies of fatty acids or their salts of general formula (1), which are used as active ingredients are included in the medicinal products.

25th

Manufacturing example for tablets - 1

Tablets containing the following compositions were prepared:

Palmitoleic acid 140 mg

Strength 31.4 mg

Lactose 125 mg

Polyvinylpyrrolidone 1.8 mg

Magnesium stearate 1.8 mg

total 300 mg

Manufacturing example for tablets - 2
10

Tablets were prepared as in the previous example, but using palmitic acid instead of palmitoleic acid or undecylenic acid was used.

Example of the production of film-coated tablets - 1

Film-coated tablets of the following composition were produced:
20th

Palmitoleic acid 150 g

Avicel 40 g

(Trade name for microcrystalline
Cellulose manufactured by Asahi
Chemical Industry Co., Ltd.)
Cornstarch
Magnesium stearate
Hydroxypropyl methyl cellulose
Polyethylene glycol 6000
0 castor oil
Methanol

30th G 2 G 10 G 3 G 40 G 40 G

Palmitoleic Acid, Avicel, Corn Starch, and Magnesium Stearate were mixed together and wedded. Then the mixture was molded into tablets using a punch with a diameter of 10 mm. The tablets were then coated with a film coating agent consisting of hydroxypropylmethyl cellulose, Polyethylene glycol 6000, castor oil and methanol to give film-coated tablets coated.

Example of the production of film-coated tablets - 2

Film-coated tablets of the following composition have been produced:

Undecylenic acid 150 g

Citric acid 1.0 g

Lactose 33.5 g

20 Calcium secondary phosphate 70.0 g

Pluronic F-68 30.0 g

Sodium lauryl sulfate 15.0 g

Polyvinyl pyrrolidone 15.0 g

Polyethylene glycol (Carbowax 1500) 4.5 g

Polyethylene glycol (Carbowax 6000) 45.0 g. .

Corn starch 30.0 g

Dry sodium lauryl sulfate 3.0g

Dry magnesium stearate 3.0 g

Ethanol. sufficient amount

undecylenic acid, citric acid, lactose, secondary calcium phosphate, Pluronic F-68 and sodium lauryl sulfate were

mixed together. The mixture was sieved through a No. 60 sieve and then using an ethanolic Solution containing polyvinylpyrrolidone, Carbowax 1500 and Carbowax 6000 granulated. The mixed powder was if necessary processed into a pasty granulate by adding ethanol. Corn starch was added to the mixture and mixing was continued until homogeneous granules were obtained. The granules were then through a Sieve No. 10 sifted and placed in a bowl and 10 to

10 dried in an oven at 100 ^° C. for 14 h. The dry one

Granules were sieved through a No. 16 sieve and then with mixed dry sodium lauryl sulfate and dry magnesium stearate. The mixture thus obtained was in a Deformed tablet machine. The core portion of these tablets was then coated and then was Talc given to prevent moisture ingress. The surface of the thus treated core parts was covered with an intermediate coating layer. A sufficient number of the coating layers has been used for oral administration performed. In order to make the surface of the tablets completely round and smooth, a Intermediate coat and another coat applied. These coated tablets were then colored, until tablets with the desired color were obtained. After drying, the coated tablets were polished, so that they received a homogeneous glossy surface.

example 1
30th

The prophylactic activity in the case of blood vessel damage in hypotensive diseases has been demonstrated for palmitoleic acid,

one of the fatty acids of the general formula (1) ^ examined as an active ingredient in the medicaments according to the invention, and the effect is clearly evident from the following test results.
5

All experiments were performed using male 2 month old SHRSP. At the first try received the SHRSP on two different diets, namely Diet-I and Diet-II as shown in Table I 10 for about 10 months. Nine (9) SHRSP were used for the Diet-I and eight (8) SHRSP were used for Diet-II. Blood pressure and the occurrence of cerebral apoplexy this SHRSP was investigated.

link

Water,% by weight proteins,% by weight oils and fats,% by weight Fibers,% by weight ash,% by weight phosphorus,% by weight soluble nitrogen-free mtrium chloride, wt% thiamine, mg / 100 g riboflavin, mg / 100 g Niacin, mg / 100 g vitamin A Iü / 100 g Carotene, ncg / 100 g calcium, wt%

Table I. Diet I. Diet II 9.3 9.0 20.5 25.3 4.4 6.2 3.3 3.5 4.85 6.46 0.63 0.74 dance, wt% 59.8 49.6 1.05 0.98 1.4 1.5 1.0 0.83 11th 11th 770 1 .200 50 430 ■ 0.683 0.825

10

15 20 25 30

---- - link - - 3 410850 - 2Ö - Cobalt, ppm Copper, ppm Diet ~ i Diet II Iron, ppm -2.9 0.9 Magnesium, ppm 21.6 7.7 Manganese, ppm 214 387 Potassium, wt% 224 775 Selenium, ppm 114 89.9 Sodium, wt% 0.650 0.754 Zinc, ppm 0.53 0.33. Nitrate nitrogen (nitrate, nitric acid), ppm 0.284 0.261 Organic acid composition of proteins 52.2 57.5 Alanine, wt% 3 14th Valine, wt% Glycine, wt .-% 0.99 1.50 Isoleucine, wt% 1.03 1.27 Leucine, wt% 0.95 1.33 Proline, wt% 0.88 1.06 Threonine, wt% 1.63 2.22 Serine, wt% 1.63 1.77 Methionine, wt% 0.77 1.02 Hydroxyproline, wt .-% 1.01 1.26 Phenylalanine, wt .-% 0.38 0.51 Aspartic acid, wt .-% 0.06 0.17 Glutamic acid, wt .-% 1.00 1.23 Tyrosine, wt% 1.65 2.29 Lysine, wt%. 4.37 4.46 Histidine, wt% 0.67 0.86 Arginine, wt% 0.96 1.34 Cystine, wt% 0.67 0.39 0.52 0.73 0.22 0.26

Figure 1 shows the effect of Diet-I and Diet-II on the Blood pressure, showing the relationship between age (days) and blood pressure of the SHRSP. From Fig. 1 becomes

it can be seen that the increase in blood pressure was somewhat suppressed in the initial phase of Diet-II. Farther O% cerebral apoplexy cases occurred after 220 days from birth in the case of Diet II compared to 60% in the case of Diet I. Also with regard to the natural After death, there were signs of cerebral apoplexy in 95% of Diet I, while only 13% of Diet II were present, which clearly indicates that there was a significant difference in terms of the two diets. a Stroke.

At first, it was thought that the suppressive effect was low Diet II on the blood pressure of the different protein composition between Diet I and Diet II was to be attributed. As a result, the following experiment was performed: Eight (8) SHRSP received Diet-I, containing 1.5% by weight methionine and the relationship between the age (days) and the blood pressure of this SHRSP, who had been chronically administered methionine was investigated. The results are shown in FIG. In this experiment another group of ten (10) SHRSPs are used as controls, this being a Diet-I without methionine. The results are also shown in FIG. As can be clearly seen from FIG suppresses the sulfur-containing amino acid (e.g. Methionine) as a protein-supplying amino acid significantly increases blood pressure and reduces its occurrence a cerebral apoplexy by about 1/3. Noteworthy is the fact that the amount of Diet-I added Methionine was 1.5% by weight and that this amount was significantly greater than the difference in methionine content between diet I and diet II [0.51 - 0.38 = 0.13 (g per 100 g diet)]. From this fact it can be seen that

- 22 -

the cerebral apoplexy-reducing effect of Diet-II not solely on its protein composition Is to be traced back.

An analysis of the composition of the fats in Diet I and Diet II has been performed and the results will be published shown in Table II.

• · Table II salary (Wt .-%) Composition of the fatty acids, which in the fats contained in the feed ent Diet I. Diet II hold were » 0.3 Lauric acid 0.4. 0.2 Myristic acid 13.7 14.5 Palmitic acid 0.3 3.3 Palmitoleic acid 4.5 3.4. Stearic acid 20.7 22.0 oleic acid 52.1 41.5 Linoleic acid 0.1 0.2 Arachidic acid 5.2 4.8 Gadoleic acid 0.3 0.4 Behenic acid .0.5 0.7 Erucic acid

In the table above, the content of is different Palmitoleic acid (POA) in the two diets is quite significant, because Diet II (containing 6.2% by weight of fat) contains about 15 times the amount of POA compared to Diet-I (containing 4.5 wt% fat).

Further, in order to confirm whether or not palimoteic acid has an effect, the occurrence of cerebral apoplexy was observed to suppress or not, a Diet III prepared with a composition similar to Diet I. In the 5 Diet-III, the amino acid content was adjusted to the same level as in Diet-I, and the fat content became also adjusted to about the same level as in Diet-I. On the other hand, the content of palmitoleic acid to 0.7% by weight, that is to about 1/5 in comparison

to the 3 »3 wt .-%, which are contained in the diet II,

humiliated. The result of the composition analysis of Diet-III is shown in Table III. Continue to be the results of the analysis of the fatty acids in the fats included in Diet-III in Table IV

shown.

Table III

Part of Diet III

Water, weight percent 9.6

Proteins, wt% 19.0 oils and fats, wt% 4.1

Fibers, wt% 3.3

Ash, wt>% 6.30

Phosphorus, wt% 0.68

Soluble nitrogen-free substances,% by weight 57.7 sodium chloride,% by weight 1.18

Thiamine, mg / 100 g 2.0

Riboflavin, mg / 100 g 1.2

Niacin, mg / 100 g 13

Vitamin A, IU / 100 g 1,000

Carotene, mcg / 100 g 140 calcium, wt% 0.869

Component (continued) Diet III Cobalt, ppm 2.7 Copper, ppm 10.9 Iron, ppm 350 Magnesium, ppm 0.111 Manganese, ppm 114 Potassium, wt% 0.826 Selenium, ppm 0.35 Sodium, wt% 0.403 Zinc, ppm 58.0 Nitrate nitrogen (nitrate, nitric acid), ppm 4th

Iodnoic acid composition of proteins

Alanine, wt% 1.02

Valine, wt%. 0.83

Glycine, wt% 0.89

Isoleucine, wt% 0.68

Leucine, wt% 1.53

Proline, wt% 1.38

Bireonin, wt% 0.76

Serine, wt% .1.05

Methionine, wt%. 0.39

Hydroxyproline, wt% 0.06

Phenylalanine, wt% 0.90

Aspartic acid, wt% 1.70

Glutamic acid, wt% 3.68

Tyrosine, wt% 0.65

Lysine, wt% O, C8

Histidine, wt% 0.47

Arginine, wt% 0.62

Cystine, wt% 0.30

Table IV

Composition of fatty acids in the content (% by weight)

Diet III Forage Fats

. Lauric acid 0.1

Myristic acid 0.9

Palmitic acid 13.7

Palmitoleic acid 0.7

Stearic acid 3.2

oleic acid 23.7

Linoleic acid 48.6

Arachidic acid 0.2

Gadoleic acid ■ 5.0

Behenic acid 0.4

Erucic acid 0.4

In this experiment, a 1% aqueous sodium chloride solution was used as drinking water to complete the test period

20 abbreviate. SHRSP (10 rats for each group) were used with the respective diets, i.e. Diet-I, Diet-II and Diet-III fed and received an aqueous 1% sodium chloride solution as drinking water. It was found that the Blood pressure rose sharply and also an increase in occurrence

25 cerebral apoplexy as shown in Table V.

Table V

Trial period Appear of cerebral apoplexy Diet III 30th Diet I. Diet II 39%
77% After 6 weeks
After 10 weeks 63%
79% 0%
61%

Cerebral apoplexy occurred 6 weeks after the administration of the diets as follows: Diet-I (most: 63%), Diet III (medium: 39%) and Diet II (lowest: 0%). After 10 weeks, the incidence of cerebral apoplexy increased to 79% in Diet I, 77% in Diet III and 61% in Diet II. These results are shown in FIG as the relationship between the incidence of cerebral apoplexy and survival time (days). The summit regarding the occurrence of cerebral apoplexy, joined first in Diet I, then in Diet III, and lastly Diet II one. This indicates that palmitoleic acid (POA) suppresses the damage in the brain vessels without one To influence the suppression of blood pressure.

To confirm the indication shown here, a Diet IV, containing 20% by weight of total fats, prepared so that it contained 44 times the amount of POA with respect to Diet-I by Milk fat containing about 3% by weight POA was added to Diet-I, and then it was examined whether Diet-IV May or not effectively suppress damage to cerebral vessels. When carrying out this experiment was usual tap water given as drinking water. A test group of 14 SHRSP received Diet-I and the other .Test group of 20 SHRSP received Diet IV. The results are shown in Figs. 4 (a) and 4 (b). It there was no difference in the increase in blood pressure between Diet I and Diet IV. Cerebral apoplexy occurred in approximately 83% in the animals that received Diet-I, as by observation of the whole natural death was noted, while it occurred in only about 5% in the animals receiving the Diet IV

- 27 -

had received. The survival time of the SHRSP was at the Diet IV fed animals, averaging 325 days, noticeably longer compared to the 306 days of the piät-I fed animals.

These results confirm that palmitoleic acid (POA) has a prophylactic effect in the event of damage to the hypertonic blood vessels, this activity being of POA was previously unknown.

Example 2

(1) SHRSPs each 3 months old were divided into five groups as shown in Table VI. Every Test group received a diet FR-2 (manufactured by Funahashi Nojo K.K.) and an 18% sodium chloride aqueous solution as drinking water. The test animals were examined with the passage of time, and the survival rate was ascertained. All test groups showed no difference in the intake of feed and drinking water.

Test group

Table VI

Number (n) of SHRSP

Fatty acids

Attempt - 1 n = 11 No encore control n = 9 Palmitoleic acid Test group A Attempt - 2 n = 10 No encore control n = 10 undecylenic acid Test group B n = 10 Palmitic acid Test group C

Amount of fatty acid added to the diet (% by weight)

The results of Trials -1 and -2 are shown in Figures 5 and 6, respectively. It has been confirmed that palmitoleic acid, Undecylenic acid and palraitic acid, which are all active components of the fatty acids of the general formula (1) according to the invention, have a very strong prophylactic activity in cerebral apoplexy.

(2) In experiment (1) above, the changes in the systolic blood pressure from SHRSP in both of the two. Control groups as well as test group A in the course determined by time (days). The results are shown in Table VII.

Table VII 15

Test group 0 days 10 days 23 days 38 days e

Control 198.9 + _4.7 201.1 + 5.5 187.5 + 6.6 (244.0 ^ 36.4)

n = 11 n = 9 'n = 3 n = 3

Test group A 199.3 + 6.6 201.3 + 8.5 189.0 + 4.0 196.7 + 6.6

20 n = 9 ri = 9 'n = 7 n = 6

(Mean + ^ standard error)

There was no difference in blood pressure in the SHRSP used as control group and test group A. were used. One cannot conclude from this that palmitoleic acid reduces blood pressure and thereby reduces the Cerebral apoplexy is suppressed.

(3) Spontaneous Hypertensive Rats (SHR) and Wister Kyoto rats (WKY), which are the maternal ancestors of SHR, were separated with a diet (MF: manufactured from Oriental Yeast Co., Ltd) containing 1% by weight palm

toleic acid, nourished for 4 weeks. Then the content of palmitoleic acid in the plasma and in the thoracic aorta and the platelet aggregation activity was measured.

Fatty acid analysis

The thoracic aortas were each removed from the examined SHR and WKY and placed in a homogenizer Glass together with 19 times the amount of an aqueous 1.15% KCl solution given. The thoracic aorta was homogenized to obtain a homogenate with a Concentration of 5%.

0.1 ml of plasma or tissue homogenate was placed in a centrifuge tube provided with a stopper, and 20 micrograms of pentadecanoic acid was added as an internal standard. Furthermore, 3 ml of a chloroform-methanol mixture (2: t) were added and then a deproteinization and extraction of the total lipids were carried out. After centrifugation at 3000 rpm for 10 minutes, the supernatant liquid was transferred to a test tube with a screw cap. The solution was concentrated to dryness at 40 ⁰ C or a lower temperature under a nitrogen gas stream. To the residue thus obtained, 0.2 to 0.25 ml of a 5% HCl / methanol solution (manufactured by Tokyo Kasei Kogyo Co., Ltd.) was added. After sealing the test tube containing the mixture, the

Mixture subjected to transmethylation on an aluminum block at 100 ^° C. for 2 h. After the mixture had cooled to room temperature, 1.0 ml of η-hexane were added,

and the mixture was stirred thoroughly to release the methyl ester to extract the fatty acids into the hexane layer. Then 1 ml of water was added and the mixture was stirred gently and then the mixture was centrifuged at 3000 rpm for 5 minutes. The hexane layer of the upper part was in transferred to another test tube and the layer was dried under a nitrogen gas stream. Before performing the analysis, the residue was redissolved in 0.1 ml of hexane and part of the resulting solution (1 to 3 Microliters) were used as a sample for gas chromatography under the following conditions:

Gas chromatography:

Device: Shimadzu GC-4M (FID)

Column: 3% Silar 10C on Cromo-

sorb W-HP 80-100 mesh, 3 mm 2 mm glass column Temperature: inlet and sensor: 280 ^° C

Column: 160-220 ⁰ C at 5 ° C / min

Carrier gas: nitrogen gas 50 ml / min

Proof: FID

Integrator: YHP 3390A

The results are shown in Table VIII.

• * Stearic acid ■ Table VIII SHR. cnuro ^ e POA admin
enough WKY ' POA administered aorta . SHR POA admin
enough 'WKY POA administered Palmitoleic acid • plasma 1.70 5.73 "Control 4 90 control
n = 5 4.68 control
n = 5 ¹ 3, «3 oleic acid (0.22) (0.61) 2.03 (0.53) 2.08 ' (0.61) 2.00 (0.4D 1.22 1.27. (0.27) 1.46 (0.30) 65.83 (0.60) 59.33 POA concentrate. ■ linolenic acid ■ ' (0.07) 1.72 (0.12) ^ 7.35 (7Λ0) 53.41 (6.62) (mg / dl plasma) (0.08) (5.13) (R, 3D Total fatty acid concentration
X. j _ ^ _ «/-.-_ / __" i \ ArachidonsMure tion (mg / nu.) 22.59
(0.4-5) 22.15 20.26 26.10 22.93 Composition of the- total Dccosakhexaenoic acid - 1.39 (0.71) 21.37 (0, H9) 27.75 (0 » ¹ +! +) 25.36 (Oj53) fatty acids (%) (0.13) V, 52
(0.1 + 0) (O? 33) 3.35 7.09 (Ό, '63) 6.47 Palmitic acid '■ · 7.31 6.68 1.17 (0.19) S35 . (0.33) 3.48 (0.18) (0.19) (0.12) 7.97 (0.37) 6.69 (0.52) 7.39 13.15 I ¹ +, 01 7.36 (0.1 + 2) 6.9? (0.19) 8.02 (0.23) (0.1 + 9) (0.36) (0.26) 31.60 (0.21+) 28.39 (0.58) 27.77 30.31 31, p5 11.47 (0.38) 28-0 (0.75) 28.28 (0.68) (0.7 ¹ O (1.42) (0.1 + 7) 28.22 (0.8I ₊ ) 22.62 (1.21) 26.3R I ¹ +, 56 12.70 28.93 (0.85) 22.1 + 0 (3.01) 25.33 (1.33) (0.96) (0.21) (0.56) 19 34 (0.89) 2.04 (3.17) 2.20 ^ 59 4.32 38.52 (l'03) 2.69 (0.19) 2.37 (0.34) (0.19) (0.2R) (1 ', 2I) 3.95 (0.12) 1.12 (0.28) 0.98 4.30 (0.51) Ij 25 (0.12) 1.06 (0.16) (0.09) (0.94) (0.14)

(Mean + standard error)

The concentration ratio of palmitoleic acid (POA) to the concentration of total fatty acids in plasma was noticeable higher in the groups that received POA than in the control groups. This shows that POA was well absorbed. The ratio of the concentration of POA to the concentration of total fatty acids in the thoracic aorta was also noticeably higher in the group that received POA than in the control groups, and the POA concentration in the thoracic aorta was higher than in plasma. This indicates that POA has a greater affinity for the aorta and that its prophylactic Effect on cerebral apoplexy due to the reinforcing activity related to blood vessels POA is due.

15 '

Platelet aggregation activity

The platelet aggregation activity was determined according to the turbidity test by Born (Nature: Volume 194, 927 (1962)), the is used extensively in the evaluation of platelet aggregation inhibitors.

The blood sample was taken from a vein and then part of the blood (9) was treated with a 3.8% trisodium citrate solution (1) mixed. The mixture of this blood sample was centrifuged at 200 G for 15 min, whereby a supernatant liquid fraction of platelet rich plasma (PRP) was obtained. The remaining sediment fraction was further centrifuged at 800 G for 10 min, leaving a supernatant liquid as low-platelet Plasma (PPP) received. The number of platelets contained in the PRP became so by diluting it with PPP set so that one received a PRP sample containing 400,000 platelets per μΐ.

- 33 -

The platelet aggregation was measured with an aggregometer (manufactured by Nikoh Bio-Science K.K.). 200 μl of the PRP sample were placed in a cuvette and leave in the aggregometer for 1 min at 37 ° C. Then ADP, collagen or Ca-ionophore became A23187 (manufactured by Sigma Co.) was added to each of the PRP samples as a platelet aggregation initiator. The aggregation was determined by the average, and the maximum aggregation rate (%) was determined according to the calculated using the following formula:

A ₃ - A
Aggregation rate (%) = χ 100

This means: A ₁ : Average PRP

A ₂ : Average of PPP

A ₃ : Permeability of PRP, the one

Aggregation initiator had been added

The results are shown in Table IX.

Table IX

control 1, 25th Maximum ADP Aggregation rate 5.0 i-H: 2, (% ■) KoI lay 0 ρ g / m: A23187 POA administered , 31, 2+> 2.5 I'M 6 ^, 2 + 8, + · 62, 5 l'g / rol _Oi W, k ' L 2x10 "^ M I control 5 + 7 μ Μ Ι 53.2 + 9. .71 » ^ 135.5 • 0 + 5.2 , 63.8 + 35.8 ; POA administered 8 + 3 .2 '■ 3; I.
! 72.6 + 65! '.80, 2 + 16.0 • 78,
• V + 3.1 ; 67.8 + 38.0 '.5O, 313 65.6 + 1. ; ⁷³ ' 0 + 0.0 *
j 81, 8 + S7 j 80.0 + 1.6 SHR '. .1' _: 63.6 + 7. 9 ^: .76,
*
• I80.5 + 5.3 .9 2

(Mean + standard error)

From the results in Table IX it can be seen that there is no difference between the experimental groups, those POA had been administered and the control groups, as seen by ADP, collagen and Ca-ionophore A23187, respectively induced platelet aggregation. This indicates that the prophylactic effect of palmitoleic acid is on the cerebral apoplexy is not due to suppression of platelet aggregation.

acute toxicity

All fatty acids or salts of the general formula (1), which are used as active ingredients in the medicaments according to of the present invention are used have low toxicity. This is shown in the following examples shown.

Undecylenic acid (rats, po): 2.5 g / kg. Palmitic acid (mice, iv): 57 ^ 3.4 mg / kg

Claims (8)

PATENT AND LAWYERS PATENTANWÄLTE D1PL.-ING. W. EITLE. DR. RER. NAT. K. HOFFMANN - DIPL.-ING. W. LEHN D1PL.-ING. K. FOCHSLE. DR. RER. NAT. B. HANSEN. DR. RER. NAT. H -A. BRAUNS DIPL.-ING. K. GORG DIPL.-ING. K. KOHLMANN · LAWYER A. NETTE 40 014 o / sm Otsuka Pharmaceutical Co., Ltd. TOKYO / JAPAN Medicines for strengthening blood vessels and their use Claims Medicinal product for strengthening blood vessels, characterized in that it is active Component of at least one fatty acid or a salt thereof of the general formula (CH> / (CKp) -COOH CnnC wherein the carbon-carbon bond of the formula a single bond or double bond (cis form), m denotes 0 or 6 and η 7 or 8 and one pharmaceutical contains acceptable carrier. ! ABELLASTRASSE < . D-8O0O MUNICH 81 ■ TELEPHONE CO8S0 O11O87 · TELEX 8-2ββ1β CPATHEJ. TELECOPER Ο183οβ 2. Medicament according to claim 1, characterized in that _{it is} in solid form
is present. 3. Medicament according to claims 1 or 2, characterized
characterized that the fatty acid
is cis-9-hexadecenoic acid (palmitoleic acid). 4. Medicament according to claims 1 or 2, characterized
characterized in that the fatty acid is 10-undecenoic acid (undecylenic acid). 5. Medicament according to claims 1 or 2, characterized g e indicates that the fatty acid is n-hexadecanoic acid (Palmitic acid) is. 6. Use of the medicament according to claims 1 or 2 as a prophylactic or therapeutic for hypertensive Vascular damage. 7. Use according to claim 6 for prophylaxis or therapeutic Treatment of cerebral apoplexy. 8. Use according to claims 6 or 7, wherein the fatty acid is palmitoleic acid, palmitic acid or undecene acid is.