DE3000139A1 - THE USE OF GLYZERINE PHOSPHORYL DERIVATIVES IN THE THERAPY OF DISLIPAEMIA, HEPATITIS AND COMPARISON-APPROPRIATE PHARMACEUTICAL COMPOSITIONS - Google Patents

Description

Dr. Ing.E. Liebau
Patent attorney (1935-1975)

- 4 patent attorneys

LIEBAU & LiEBAU

Birkenstrasse 39 D-8900 Augsburg 22

Oipl. Ing. G. Liebau patent attorney

Patent attorneys Liebaui. Liebau ■ Birkenstrasse 39 ■ D-89 00 Augsburg

Telephone (0821) 96096 cables: elpatent augsburi

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your / your ref.

LPB Istituto Farmaceutico s.p.a.

Via dei Lavoratori,

20092 Cinisello Balsamo (Milano) Italy

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Date:
date

The use of glycerol phosphoryl derivatives in the therapy of dislipemia, hepatitis and the like pathological conditions and pharmaceutical compositions suitable therefor.

The invention relates to the use of glycerol phosphoryl derivatives of the general formula (I)

CH ₀ OH

I ²

CHOH CH ₀ O-

(D

R where R is H or COOH and R ^{1 is} H or CH ₃ .

According to the invention is the use of glycerol phosphoryl derivatives of the general formula (I)

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Bank details. Post office in Munich, account 86510-809 h '0010080 Dt utsche Bank AG Augsburg. Account 0834192. BLZ 72070001

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CH ₀ OH
ι «a

C.
1

HIGH

+
L ₂ -CH-N (R ¹ ),

where R stands for H or COOH, R 'stands for H or CH ₃ , intended for the treatment of dislipemia, alcoholism, acute, subacute and chronic hepatitis, steatosis and comparable pathological conditions =

In particular, the pharmaceutical compositions according to the invention contain at least one active principle the compounds mentioned below;

Glycerine phosphorylcholine (R - H, R '= CH ₃ ), glycerine phosphorylethanolamine (R = H, R ^! = H), glycerine phosphorylserine (R = COOH, R' = H).

Otherwise it is not important for practical use which of these compounds is administered because the intermediate metabolism of humans can convert these three compounds into one another »The active principle a composition according to the invention can, for example, consist solely of glycerol phosphorylcholine or also from an association of all three connections exist.

It has surprisingly been found that the compounds of the general formula (I) are antidislipemic Have an effect (especially with regard to blood lipids and blood lipoproteins) and the metabolism in a way initiate, in which, among other things, the metabolism and the conversion of fats is favored (antisteatosis activity). Thanks to these properties and in conjunction with one

With an extraordinarily favorable toxicological profile, it is possible to use the compounds therapeutically for treatment to use certain pathological disorders.

The therapeutic usefulness of the compounds of the general formula (I) is now intended from various points of view be treated.

toxicity

It can be shown that the compounds of general formula (I) which are of particular interest - glycerol phosphorylcholine, Glycerine phosphorylethanolamine, glycerine phosphorylserine - as normal metabolic products occur in the human body, and it is therefore not acceptable to use the terms subacute and chronic in them Related toxicity.

As far as the acute toxicity of these compounds is concerned, experiments using L-OC-glycerol phosphorylcholine were employed, draw the following conclusions:

1) L-o £ -Glycerolphosphorylcholine has a toxicity, those with oral administration up to an amount of 3 g / kg) in all investigated species (mice, rats, Rabbit) neither leads to death nor shows any symptoms;

2) the lethal dose in the case of parenteral administration differs depending on the type of administration; L-oC-glycerol phosphorylcholine administered intraperitoneally resulted in an LD_ with a maximum of 1500 mg / kg in rats and mice, in rabbits of 1000 mg / kg;

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3) the LDg is ₀ when administered intramuscularly

about 1300 mg / kg in rats, about 1000 mg / kg in mice and about 600 mg / kg in rabbits «

Antidislipemia activity

The antidislipemia activity of the compounds is illustrated, for example, by the following test results:

a) Buchanan's test

Twenty male rats with an average weight of 200 g received L-oC-glycerol phosphorylcholine in doses of 100 mg / kg / day and 200 mg / kg / day by intramuscular injection on four consecutive days. The following occurred: a decrease in total blood cholesterol of 10% or 17 % , while the (^ -lipoproteins increased by 17 % or 40 % ; these results were based on the results on a further ten untreated animals. At the same time, a corresponding percentage that was to be expected was obtained Decrease in ß-lipoproteins "which, unlike σΙ-lipoproteins, are atherogenic.

b) Diet with ethanol

Twenty male rats with an average weight of 200 g and twenty female rats with an average weight of 170 g were taken over the course of four Days 100 mg / kg / day L-oC-glycerol phosphorylchlorin intraperitoneally administered »Beginning on the second day of treatment, the animals received ethanol, which was used as 10% was added to the water that was freely available? on the third and fourth day the rats also fed ethanol through a gastric tube in an amount of 0.5 g per animal

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there is a net decrease in blood triglycerides that in the males 38%, in the females 45% in comparison to the animals, which only with ethanol had been treated (ten males, ten females).

^c ) lipolysis during fasting

Three groups of male rats (each group of 20 animals) with an average weight of 195 g received intraperitoneally 1-cC-glycerol phosphorylcholine in amounts from 100 to 200 or 400 mg / kg at the beginning of a forty-eight hour fast during which they only use water could consume ad libitum. After twenty-four hours The animals were fasted an equal amount (equal to the previous one) of L-OC-glycerol phosphorylcholine administered. At the.' At the end of the fast, the rats were killed painlessly. There was a decrease in NEFA (non-esterified fatty acids), the amounts of decrease were 7% and 11% and 21%, respectively, over the Control group rats (ten for each group)

~ Pharmacokinetic studies

The activity described above can be interpreted based on the following results:

Labeled L-glycerol phosphorylcholine, which is administered intravenously, intramuscularly and orally administered was completely and rapidly reabsorbed so that the administered was labeled Material was found almost entirely in the circulatory system in all cases.

A large part (at least 50%) of the labeled administered material disappeared very quickly (in the first Hour) from the circulatory system and found itself in it

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Phase mainly in the liver (an insignificant part in the brain); during the third and fourth hours after entering, the labeled glycerol phosphorylcholine returned to the circulatory system so that it was converted into By the sixth hour, at least 80 to 90% of it was back in the circulatory system and contained in the lipoproteins was. This proves that the glycerol phosphorylcholine was mainly used for the synthesis of lipoproteins. In addition, it was in the hundredth hour After the administration in the circulatory system, the radioactivity recorded in the sixth Hour was reached.

The values obtained show that glycerol phosphorylcholine is actually an agent that intervenes in lipid metabolism and has the effect of anti-islipemia substances? the effect of this substance, which is less strong than the modern antidislipemia remedies, has the very remarkable advantage " that it is not associated with acute, subacute or chronic tossisity"

Activation of the liver metabolism

The test values below show the surprising protective effect of the compound (Σ) _t, in particular of L-oC-glycerol phosphorylcholine, compared with the toxic effect caused by carbon tetrachloride, by a fat diet, by experimental cholemia and by Q (.- naphthyl isothiocyanate.
In detail;

1. L-O ^ -Glyserinphosphorylcholine, intramuscularly in one Dosage of 100 mg / kg administered to rats treated with a single dose of CCl ^ continues the mortality rate reduced by 50%, reduced in the serum

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the increase in GOT (glutamate oxaloacetate transaminase) (- 70%), of GPT (glutamate pyruvate transaminase) (-68%) and of LDH (lactate dehydrogenase) (-72%) and shows the occurrence of histological changes to be minor In the liver area.

2. L-ot-glycerol phosphorylcholine, administered intramuscularly at a dose of 100 mg / kg to rats _{treated with a single dose of CCl 4} , interferes with the retention of BSP (bromosulfalein) in the blood, which is caused by the toxicity of CCl ₄ was caused.

3. L-oC-glycerol phosphorylcholine, intramuscularly in one Dosage of 100 mg / kg given to rats with experimentally produced cholemia (binding of the bile duct), decreases the increase in GOT (-31%) and GPT (-45%) in the serum.

4. L-o ^ -glycerol phosphorylcholine, intramuscularly in one Administered at a dose of 100 mg / kg, rats may have antagonistic activity against hepatic steatosis caused by overeating with glycides, lipids and proteins: weight the liver (expressed in g / 100 g body weight of the animal) and the amount of fat contained in the liver (expressed in g / 100 g fresh tissue) in control animals, in those treated with the unbalanced diet Animals and those animals that have an unbalanced diet with more L-oC-glycerol phosphorylcholine are listed in the table below.

Incidentally, oral administration of glycerol provides phosphorylcholine a comparable result in terms of the changes made by having a diet excessive content of glycides, lipids and proteins

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were brought about. The fat infiltration is evident lower in the animals treated with glycerol phosphorylcholine have been treated? see »the values given in the table =

L-oC-glycerol phosphorylcholine, intramuscularly in one Dosage of 100 mg / kg administered to rats that had been poisoned with oC-naphthyl isothiocyanate able to develop good antagonistic activity with death not occurring, a decrease there is an increase in GOT (-76 S), GPT (-53%) and LDH (-77%) and there is also a decrease the rise in CC-globulin and ß-globulin occurs and finally intervened to help reduce histological changes in the liver becomes (slight blood overcrowding of the sinusoid and the edema state of the portal vein spaces, slight Overgrowths associated with the internal duct of the liver and minor damage to the Liver cells) =

TABEL

Liver weight
(g / 100 g body
weight) orally Fats in the fresh
liver (g / 100 g
Fresh tissue) intra
muscular 3.50
4.37
3.61 intra-oral
muscular groups 3.40
4.26
3.79 4.16 4.22
11.90 12.60
7.55 6.18 Control group
treated with un
more balanced
nourishment
treated with un
more balanced
Nutrition plus
Glycerine phospho-
rylcholine

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Clinical tests

The pharmacological indicated by the above numerical values Activity was evident during clinical tests and reports on indications for lipid metabolism as well as through hepatological indications confirm.

¹ · Report on indications related to lipid metabolism

Antidislipoproteinic and antitriglyceridic vision agents Clearing the serus.

The clinical examination was carried out on about twenty patients / those on various forms of the Dislipemia suffered: It was shown that L-oC-glycerol phosphorylcholine a marked effect in the direction of triglyceridemia reduction and improvement de3 has the ratio of c <_- to ß-lipoproteins.

In addition, there is always a very clear effect on alcoholics.

2. Report on indications with regard to liver metabolism

Acute, subacute and chronic hepatitis steatosis.

L-gC-glycerol phosphorylcholine has special activity in acute hepatitis with cholostatic features shown in double-blind cross-experiment studies (English: cross-over to blind double), whereby not only the blood level of the transaminases but above all alkaline phosphatase and bilirubin levels were also improved.

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Glycerol phosphorylcholine has been shown to be effective against persistent chronic hepatitis; Advantages in the treatment of the initial forms of cirrhosis.

In active chronic hepatitis, glycerol phosphorylcholine has no effect "

According to the results of various toxicological, pharmacological and clinical examinations prove the compounds of the general formula (I) prove to be extremely effective for therapeutic use. For this purpose they can be put together to form compositions according to the known pharmaceutical methods are, the essential active ingredient of which is a compound of formula (I) together with a a suitable pharmaceutical carrier. If necessary, the compositions can be put together in dosage units appropriate for a particular dosage form, with the amount of active ingredient is chosen in each dosage unit so that for each therapeutic administration one or several units are required, the dosage unit For example, it can be a tablet, a pill, a pillow, a packaged powder, or one in capsules included powder for oral administration may be in the form of a suppository or a injectable sterile solution or suspension, if necessary in an ampoule, for have parenteral administration.

Below are examples of such pharmaceutical dosage units %

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EXAMPLE 1

Tablets

AWAY

L-oC-glycerol phosphorylcholine 100 mg 200 mg

Lactose 50 mg 100 mg

Strength 32.5 mg 65 mg

Talc 10 mg 20 mg

Avicel 55 mg 110 mg

Magnesium stearate 2.5 mg 5 mg

EXAMPLE 2

Tablets

Compositions were prepared according to Example 1, but the glycerol phosphorylcholine was replaced by the same Amounts (100 or 200 mg) of glycerol phosphorylethanolamine or glycerol phosphorylserine replaced.

EXAMPLE 2

Pills

AWAY

Glycerol phosphorylcholine

Lactose (or ethanolamine or serine) starch

Talc Avicel

Magnesium stearate Eudragite E magnesium carbonate titanium dioxide

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100 mg 200 mg 50 mg 100 mg 32, 5 mg 65 mg 50 mg 80 mg 55 mg 110 mg 2.5 mg 5 mg 0.5 mg 0.75 mg 4th mg 6th mg 5 mg 7.5 mg

AWAY

Kaolin 6.5 rag 9.75 mg

Gelatin 2.0 mg 3 mg

Sucrose 90 mg 135 mg

Gum arabic 2 mg 3 mg

EXAMPLE 4 What gelatin capsules

In soft gelatin capsules of a suitable shape were packed:

_JL B

Glycerol phosphorylcholine

(or serine or ethanolamine) 100 mg 200 mg

Vegetable oil 20 mg 40 mg

EXAMPLE 5 Hard gelatin capsules

In hard gelatine capsules of a suitable shape were packed:

A B

Glycerol phosphorylcholine

(or serine or ethylamine) 100 mg 200 mg

Avicel 50 mg 100 mg

Strength 18 mg 36 mg

Lactose 15 mg 30 mg

Talc 15 mg 30 mg

Magnesium stearate 2 mg 4 mg

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EXAMPLE 6 Freeze-dried ampoules

AWAY

Glycerol phosphorylcholine

(or serine or ethanolamine) 100 mg 200 mg

Glycine 250 mg 250 mg

Methyl p-hydroxybenzoate 1.68 mg 1.68 mg

Propy-p-hydroxybenzoate 0.48 mg 0.48 mg

EXAMPLE 7

AWAY

Glycerol phosphorylcholine

(or serine or ethanolamine) 100 mg 200 mg NaH ₂ PO ₄ -Na ₃ HPO ₄ (M / 10), pH 6.5 qb 1 ml 2 ml

Similar approaches result if, instead of the individual active principles, identical mixtures of two or employs more than two active principles.

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Claims (10)

Patent claims: _ 1. Use of glycerol phosphoryl derivatives of the general formula (I) H ₂ OH <ί ^Η0Η (D CH ₂ -CH-N (R ¹ ) ₃ R in which R stands for H or COOH and R 'for H or CH ₃ , in the therapy of dislipemia, alcoholism, acute, subacute or chronic hepatitis, steatosis and comparable pathological conditions ο 2. Use of glycerol phosphorylcholine in the therapy of dislipemia, alcoholism, acute, subacute and chronic hepatitis, steatosis and comparable pathological conditions. ■! 3 © 0f4 / 0912 3. Use of glycerol phosphorylethanolamine in the therapy of dislipemia, alcoholism, acute, subacute and chronic hepatitis, steatosis and similar pathological conditions. 4. Use of glycerol phosphorylserine in the therapy of dislipemia, alcoholism, acute, subacute and chronic hepatitis, steatosis and similar pathological conditions. 5. Use of a combination of at least two glycerol phosphoryl derivatives of the general formula (I) in the therapy of dislipemia, alcoholism, the acute, subacute and chronic hepatitis, steatosis and comparable pathological conditions. 6. Pharmaceutical composition for the treatment of dislipemia, alcoholism, acute, subacute and chronic hepatitis, steatosis and comparable pathological conditions, characterized in that as active principle of at least one glycerol phosphoryl derivative of the formula (I) according to claim 1 and furthermore a pharmacologically acceptable carriers are provided. 7. Pharmaceutical composition according to claim 6, characterized in that the active principle is glycerol phosphorylcholine, Glycerine phosphorylethanolamine or glycerine phosphorylserine, either individually or in combination is provided with each other. 1300U / 0912 : ": i -:.;:": 30001 8. Pharmaceutical composition according to claim 6 or 7 in a preparation suitable for oral administration. 9. A pharmaceutical composition according to claim 6 or 7 in a preparation suitable for parenteral administration. 10. Pharmaceutical composition according to one of the examples up to 7 «, 1300U / 091