DE4117629A1 - Use of phosphatidyl serine derivs. - for treatment of depression and loss of cerebral function e.g. Parkinson's disease and Alzheimer's disease - Google Patents

Abstract

Use of 1,2-substd. phosphatidyl serine derivs. of formula (I) or a salt of (I), in prepn. of a medicament for therapy of disorders leading to depression and/or disorders or cerebral functions, esp. redn. or loss of memory, is new. In (I) R1, R2 = satd. or unsatd. 8-24C hydrocarbyl residue. USE/ADVANTAGE - (I) are useful in treatment of diseases such as Parkinson's disease, Alzheimer's disease, organic brain syndrome, AIDS dementia, depressive pseudo-dementia, dementia(l) syndromes, deliria, intoxications, withdrawal syndromes or cytopathic influences. Dosage is 1-20 mg/kg/day administered, e.g. as tablets or by injection. Some phosphatidyl serine derivs. are known for treatment of tumours and CNS disorders.

Description

The present invention relates to the use of che mix defined 1,2-substituted phosphatidylserines to Production of a drug.

Symmetrically substituted phosphatidylserines are per se known. So z. B. Dioleoylphosphatidylserine an object of European Patent 00 99 068 and PCT / DE83 / 0123 or the publications P. Wooley and H. Eibl, Chem. Physics Lipids 47 (1988), 55-62.

The chemically known from the above-mentioned documents Compounds defined so far are known as pharmaceutical Excipients in the production of liposomes, which is a Arz means such as B. Muramyldipeptid, for the treatment of Tumor diseases used (see H. Eibl, C. Unger, Phos pholipids as antitumor agents: possibilities of a selective Therapy, in: The cell membrane as a point of attack of the tumor rapie, Current Oncology 34, edited by Unger et al., W. Zuckschwerdt Verlag, Munich).

A natural phospholipid isolated from animal brains pid composition, which is a mixture of several different non-chemically defined phosphatidylserines and phosphatidylethanolamines in a specified Ver contains, is already used to treat disorders of the central nervous system (CNS) used (US-PS 45 95 680). The mode of action of the phospholipid composition is based on an influence of aged membranes of the attributed to the central nervous system in which either the Relief of transmitter substances facilitated or the Signal transmission is promoted (Chang, H.W., Br.J.Pharma col. 93 (1988), 647-653 and Clinical Trials Journal, Vol. 24 (1987), No. 1). This will u. a. the at reduced brain performance of pathologically altered dopamine or acetylcholine Transmitter systems positively influenced.  

The patent application WO87 / 05 024 discloses pharmaceutical Compositions containing glycerophosphoryl-O-serine as active Contain component and on the metabolism of the brain and act on the neuronal transmission. Glycerophosphoryl- O-serine is the precursor of phosphatidylserines in the body.

The object of the present invention was to provide a ver to find a meaningful way to treat diseases which causes the onset of depressive symptoms and disorders comes from brain power and memory functions.

This object is achieved by the use of the invention tion of a chemically defined 1,2-substituted phosphati dylserines of the general formula (I)

wherein R ₁ and R _{2 are the} same or different and each represent a saturated or unsaturated, branched or geradketti gene hydrocarbon radical having 8 to 24 carbon atoms, or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of Erkran diseases in which it comes to depression and / or disorders of brain function functions, in particular to relaxation or loss of memory functions.

The term "chemically defined" within the meaning of the present Invention is intended to exclude mixtures of phosphatidylserines, how they are isolated from the brain of animals. The chemi synthesis of symmetric or unsymmetrical phospha tidylserinen is known, see, for. Eibl and Woolley, Chem. Physics Lipids 41 (1986) 53-63; Eibl and Woolley, Chem.Phy  sics Lipids 47 (1988), 47-53; Wolley and Eibl, Chem.Physics Lipids 47 (1988), 55-62 and Eibl and Woolley, Chem. Physics Lipids 47 (1988), 63-68.

In the formula (I), the radicals -OCO-R₁ and -OCO-R₂ are, for example Caprylic, capric, lauric, myristic, palmitic, Stearic, oleic, linoleic, cis-11-eicosenic, erucic or neuronic acid residues.

The radicals R ₁ and R ₂ in the general formula (I) may be the same or different. However, particular preference is given to those compounds in which the radicals R ₁ and R _{2 are} identical, ie symmetrically substituted phosphatidylserines. In this sense, compounds in which R ₁ and R ₂ are not identical but are identical, ie belong to a class of hydrocarbon radicals (either the saturated or the unsaturated hydrocarbons), are also preferred in this sense. Among these, in turn, those compounds are preferred in which R ₁ and R _{2 are} mono- or polyunsaturated, geradket term hydrocarbon radicals. It is particularly preferred if R ₁ or / and R _{2 are} oleic acid residues. Most preferred are pharmaceutical compositions containing synthetic and substantially pure sodium 1,2-dioleylphosphatidyl- (S) serine.

It is known from the prior art that the use of Phosphatidylserine mixtures as found in brain extracts of Animals are obtained (US Patent 45 95 680), or syn thetic precursors of phosphatidylserines, eg. B. glycero phosphoryl-O-serine (WO 87 05 024) in certain animal models an enhancing effect on brain performance, memory or exercise the spatial orientation. Therefore, na natural phosphatidylase isolated from animal brains rin mixtures already occasionally used clinically. Your Mode of action is based on an influence of aged Returned membranes of the CNS, so that either the Freiset  tion of transmitter substances or the signal transmission is encouraged. In particular, the at reduced brain performance pathologically altered dopamine or acetylcholine Transmittersysteme positively influenced.

According to the present invention has now been surprisingly found that chemically defined, symmetrically 1,2-sub substituted phosphatidylserines of the general formula (I), in particular dioleylphosphatidylserine, in animal models, the for the development of brain-enhancing and antidepres effective substances, an improvement of the brain performance and memory functions as well as an antidepressive Show an effect on the previously known phosphatidylserine Extracts or glycerophosphoryl-O-serine significantly superior are.

It should be noted that an antidepressant agent profile in a prior art means, d. H. the Brain extracts or the glycerophosphoserine, experimental is not tangible. In the inventive chemically defi nated, symmetrically 1,2-substituted phosphatidylserines the general formula (I), however, can be an antide statistically significant evidence of pressive effect.

The results of the animal experiments can be expected that the use according to the invention of chemically defined, symme trically 1,2-substituted phosphatidylserines of the general Formula (I) is suitable for the therapy of diseases, in which leads to a loss of brain power and memory functions comes, z. B. in the course of pathological old processes such as Parkinson's disease, Alzheimer's disease, the brain organic psychosyndrome (Organic Brain Syndrome), AIDS dementia, depressive pseudo-dementia, dementia Syndromes, delirium as acute organic brain syndromes, Intoxications, withdrawal syndromes or cytopathic effects sen.  

Accordingly, the present invention also relates to a medicament agent for the treatment of diseases in which it is too Depression and / or disorders of brain function, in particular to easing or loss of memory radio arrives, which is characterized in that it as Active substance one or more chemically defined, symmetrical 1,2-substituted phosphatidylserines of the general formula (I)

wherein R ₁ and R _{2 are the} same or different and each represent a saturated or unsaturated, branched or geradketti gene hydrocarbon radical having 8 to 24 carbon atoms, or a pharmaceutically acceptable salt thereof, if appropriate together with pharmaceutically customary carrier, auxiliary, Contains fillers and diluents. Preferably, R ₁ and R _{2 are the} same or similar. It is further preferred if R ₁ and R _{2 represent} mono- or polyunsaturated, straight-chain hydrocarbons.

Examples of medicaments according to the invention are those which synthetic, substantially pure sodium 1,2-dioleoyl phosphatidyl (S) -serine or sodium 1,2-dipalmitoyl-phospha tidyl (S) -serine as an active ingredient.

The compounds of formula (I) are preferably in Dosie from 0.01 to 100 mg / kg body weight / day. Suitable dosage forms are z. B. Combinations of active substance with common pharmaceutical carrier, Auxiliaries, fillers and diluents in the form of tablets,  coated tablets, dragees, suppositories, semi-solid Formulations, liposomes, emulsions, sterile solutions or Suspensions for injection. Pharmaceutically usable Carriers are z. As lactose, sucrose, sorbitol, talc, sterol acid, magnesium stearate, lipids, gum arabic, cornstarch or cellulose, in combination with solvents such as water, Polyethylene glycol etc.

The medicaments according to the invention contain a Thera pie of disorders that cause depression or / and Disturbances of brain power and memory functions come sufficient amount of active ingredient, optionally together with pharmaceutically customary carrier, auxiliary, fillers and diluents planning agents. For example, such drugs contain 0.01 to 98 wt .-% of at least one inventive Active substance together with at least one pharmaceutical Carrier.

The method of treating diseases in which it to depression or / and a loss of brain power and Memory functions includes the administration of a therapeutically effective amount of active substance on a Patient in need of this treatment.

The dosage of the medicament according to the invention depends in first line of the specific dosage form and of the Purpose of the therapy. The size of the single doses and the Administration scheme can best be determined by an indivi dulichen assessment of each case by the Doctor to be determined, taking the age, weight and the Condition of the recipient, the route of administration and the type and Severity of the disease must be considered. virtue example, the daily dose is 1 to 20 mg / kg body weight.

The duration of the treatment depends on the type and severity the disease. It will generally be over several  Weeks, for example 4 to 8 weeks. If that Present in the form of a metering unit, this contains preferably 10 to 500 mg of the invention for use coming connection.

The medicaments of the invention may be oral or parent administered. Oral administration may be in fe ster form, z. As tablets, lozenges, capsules, powders or in liquid form, e.g. B. as aqueous or oily suspensions ions, emulsions, liposomal preparations, syrups, elixirs, Solution or filled with liquid capsules. before Oral remedies are in the form of tablets or capes and conventional supports such as binders (eg. Syrup, acacia, gelatin, sorbitol, tragacanth or polyvinyl pyrrolidone), fillers (eg lactose, sugar, corn starch, Potato starch, calcium phosphate, sorbitol or glycine), slip (for example, magnesium stearate, talc, polyethylene glycol or Silica), disintegrating agents (e.g., starch) and mesh medium (eg sodium lauryl sulfate).

Agents for parenteral administration are generally in the form of a solution or suspension of the invention for Application coming compound along with usual pharma zeutischen carriers before, for example in the form of an aqueous Solution for intravenous injection, a liposomal preparation tion or an oily suspension for the intramuscular Injection. Means suitable for parenteral administration is obtained by 0.1 to 10 wt .-% of the inventive Active ingredient in water or a carrier made from an alipha Polyalcohol such as glycerol, propylene glycol or poly ethylene glycol or a mixture thereof dissolves. The Polyethylene glycols consist of a mixture of non-volatile ger, usually liquid polyethylene glycols, both in Water as well as in organic liquids are soluble. The molecular weights of suitable polyethylene glycols lie preferably in a range of from 200 to 1500 enough.  

Pharmaceutical agents for rectal administration are in Form of suppositories, the inventions use verwen compounds in a suitable suppository base ge, such as. As cocoa butter, hydrogenated fats, poly waxes or Polyethylene glycols in an amount of 1 to 1 10 wt .-% are included.

The preparation of the medicaments according to the invention takes place by conventional methods, for example by tableting, Incorporation of the present invention used Compounds in a suppository base, sterile filtration and filling in ampoules or dropper bottles of a solution of in accordance with the invention used in Injection water along with common additives, such as sodium chloride, sodium hydrogen phosphate, disodium EDTA, benzyl alcohol hol or sodium hydroxide to adjust the pH.

The following examples serve to illustrate the inventions dung.

example 1 Pharmacological-toxicological behavior profile in mice

Experimental procedure:
All substances are tested in standard doses 4, 12, 36, 108 and 324 mg / kg. The highest dose (324 mg / kg) is dissolved or suspended in 1% methylcellulose + 1% Tween 80. From this, the lower dosages are obtained by dilution Ver. The experimental animals used are female NMRI mice (breeders: Charles River Wiga) weighing 18 to 25 g and 5 animals per dosage, ie a total of 25 animals. The application volume is 10 ml / kg and the type of administration is intraperitoneal. The 5 lowest dose animals are administered first; afterwards, in the course of 10 minutes, the next higher dosage is applied, etc.

Test parameters:
Lokomotorik and exploratory activity were determined according to S. Irwin (Psychopharmacologica 13, 222-257 (1968)) in the open-field cage. From the score values, ED ₅₀ values were calculated according to Litchfield and Wilcoxon (JT Litchfield and F. Wilcoxon, J. Pharmacol. Exp. Ther 96, (1949), 99-113)).

The evaluation of the 5 animals of one dose takes place 40 min. reagent grade

Tested substances:
There are the inventive chemically defined Phos phatidylserine (PS) 1,2-dioleoyl-PS and 1,2-dipalmitoyl-PS with lecithin and a natural, from animal brains gewonne NEN phosphatidylserine extract according to US-PS 45 95 680 vergli chen.

Results

It can be seen that only 1,2-dioleoyl-PS is the only app tested test substances with a score of 16.2 Lokomotorik and exploratory activity of mice very clearly influenced. Here are the differences noted statistically to other substances at the 0,05% level significant. It has also been found that dipalmitoyl-PS as 1,2-symmetrically substituted with saturated fatty acids synthetic PS, a qualitative and quantitative with the PS extract from animal brains has comparable effect.  

Example 2 Telemetric derivations of electroencephalographic effects on freely mobile rats

Experimental procedure:
The experiments were carried out as described by Dimpfel et al. (Neuropsychobiology 18 (1987), 212-218)). The animals received the test substances in a volume of 1 ml / kg body weight intraperitoneally.

The dose was 50 μmol / kg in each case. Control animals received physiological saline.

EEG spectra will be 15 minutes before and 50 minutes after Substance application registered.

Test parameters:
Substance effects were determined as a percentage change in the spectral power density of the EEG spectra in the α-band (7.5-9 Hz) of the frontal cortex relative to the values before substance application (multivariate F-test).

Tested substances:
In addition to the substances used in Example 1, additional glycerophosphoryl-0-serine (GPS-Ca ²⁺ ) according to WO87 / 05024 was also investigated.

Results

This series of studies shows that after treatment the Rats with equimolar doses of the test substances only 1,2- Dioleoyl PS to assess a vigilanzsteigernde effect significant effect (see W. Dimpfel et al. supra). 1,2-Dipalmitoyl-PS is qualitatively and quantitatively natural, chemically undefined PS extract comparison bar.

Example 3 Antagonism of mouse reserpine hypothermia (Antide pressiva test)

Experimental procedure:
Hypothermia was induced in mice by subcutaneous administration of 2.5 mg / kg reserpine. After 18 hours, the test substances were administered intraperitoneally in a volume of 10 ml / kg. The dose was 50 μmol / kg body weight. Subsequently, the rectal body temperature was recorded over a period of two hours. The experimental and control groups each comprised 10 animals.

Test parameters:
Increase in the rectal body temperature of the experimental animals compared to the control (physiological saline).

Results

Temperature difference (° C)

It can be seen that, in contrast to GPS Ca ²⁺ in equimolar concentration, 1,2-dioleoyl-PS affects the body temperature of mice to about the same extent as the standard antidepressant desipramine. The differences are statistically significant at the 0.05% level.

example 4 Various administration forms of the medicament according to the invention

Example 4a tablet formulation Active substance | 10 mg lactose 18 mg potato starch 38 mg gelatin 2 mg talc 2 mg magnesium stearate 0.1 mg

Example 4b tablet formulation Active ingredient | 50 mg potato starch 45 mg polyvinylpyrrolidone 5 mg

The tablets can be made with a colored sugar layer or Coated dragee cover.  

The dragee cover can consist of:

Sugar | 65.0 mg talcum 39.0 mg calcium carbonate 13.0 mg gum arabic 6.5 mg corn starch 3.7 mg shellac 1.1 mg Polyethylene glycol 6000 0.2 mg Magnesia usta 1.3 mg dye  0.2 mg 130.0 mg

Total Dragee Weight at 50 mg Core = 180 mg

Example 4c capsule formulation Active ingredient | 50 mg corn starch 90 mg lactose 50 mg talc 2 mg

This mixture is wrapped in gelatin capsules.

Example 4d Liquid oral formulation Active ingredient | 2 g sucrose 250 g glucose 300 g d-Sorbitol 150 g Agar Agar 0.15 g methylparaben 0.5 g Propylparaben 0.05 g Flavor (orange flavor) 10 g Tartrazine yellow 0.05 g Purified water on 1000 ml

Example 4e Liquid oral formulation Active ingredient | 2 g tragacanth 7 g glycerin 50 g sucrose 400 g Methylparabren 0.5 g Propylparabren 0.05 g Flavor (blackcurrant flavor) Red Dye No. 2C.E.184 0.02 g Purified water on 1000 ml

Example 4f Liquid oral formulation Active ingredient | 5 g sucrose 400 g Tincture of bitter orange peel 20 g Tincture of sweet orange peel 15 g Purified water on 1000 ml

example 4g injection

To prepare a 0.5% solution, 0.5% active ingredient and 0.8% sodium chloride DAB 9 in bidistilled water solved. The solution is passed through a degerming layer filter filtered, filled in 2 ml ampoules and at 120 ° C in the car sterilized.  

Example 4h infusion

To prepare a 0.01% solution for infusion 0.01% active ingredient and 5% laevulose in bidistilled water solved. The solution is filtered through degermination filters, in 500 ml infusion bottles filled and sterilized.

The example refers to 50 mg of active ingredient per single dose sis.

Example 5 Synthesis of phosphatidylserine

1) Symmetrical phosphatidylserines
1. Phosphorylation:
1,2-Dibenzylglycerol is dissolved in tetrahydrofuran (THF) (0.1 mol in 100 ml of THF). The solution is added dropwise and with stirring to a mixture of phosphorus oxychloride (0.12 mol) and triethylamine (0.18 mol) in THF (100 ml) in an ice / water bath so that the temperature does not exceed 15 ° C , The phosphorylation is completed immediately after the dropping, as a thin-layer chromatographic test shows.

2. Phosphorylation:
The resulting phosphoric acid dichloride of 1,2-dibenzylglycerol is then added dropwise with another alcohol, N-BOC-serine tert-butyl ester (0.15 mol), triethylamine (0.20 mol) in THF (200 ml) 25 to 30 ° C offset. The reaction is complete after about 3 hours (Dünnschichtchromatogra phie). The obtained 1,2-dibenzyl-sn-glycero-3-phosphoric acid N-BOC-serine tert-butyl ester monochloride is stirred for 3 hours after the addition of H ₂ O (100 ml) and thereby hydrolyzed. It is combined with CHCl ₃ (500 ml), H ₂ O (800 ml) and CH ₃ OH (800 ml), shaken and the lower chloroform phase is separated off. The mixture is again extracted with 300 ml of CHCl ₃ and the combined chloroform phases are concentrated on a rotary evaporator. The residue is taken up in CHCl ₃ / CH ₃ CH / H ₂ O 65/35/5 (ml) and passed through a short column of 200 g of silica gel. Fractions containing product are collected, mixed with 10 g of Pd / C catalyst and shaken in a hydrogen atmosphere until H ₂ uptake is complete. The solvent is finally removed on a rotary evaporator with toluene until no more water separation takes place and takes the residual oil in dimethylformamide (400 ml).

acylation:
The product, sn-glycero-3-phospho-N-BOC-serine tert-butyl ester in DMF (400 ml) is stirred at 25 ° C and treated dropwise with a solution of acyl chloride (0.12 mol), triethylamine ( 0.15 mol) and 4-dimethylaminopyridine (0.02 mol) in THF (400 ml). The acylation is completed after about 5 hours. The reaction mixture is combined with CHCl ₃ (500 ml), water (800 ml) and methanol (800 ml), shaken and, after phase separation, the lower CHCl ₃ phase is separated off. It is back-extracted with CHCl ₃ (300 ml), and the combined chloroform phases are freed from the solvent in vacuo.

Cleavage of the protecting groups:
Protected phosphatidylserine (0.1 mol) is dissolved in 1 L CH ₂ Cl ₂ , treated with 1 L trifluoroacetic acid and cooled to 10 ° C. With stirring, it is mixed with 500 ml of 70% perchloric acid. After 30 minutes at 10 ° C., the solution is mixed with 4 l of H ₂ O, 2 l of CHCl ₃ and 4 l of CH ₃ OH in the order mentioned. After phase separation, the lower phase is shaken with 2 l of 0.5 M sodium carbonate, treated with 2 l of CH ₃ OH, shaken again and separated after phase separation, the lower phase. The solvent is removed in the lower phase in vacuo and the residue is combined with 2 l CHCl ₃ / CH ₃ OH 1: 2 (ml). The crystals are filtered off with suction and washed with the above mixture.

Structures of products using appropriate Acyl chlorides are produced:

1,2-dilauroyl-sn-G-3-PS C₁₂ 1,2-dimyristoyl-sn-G-3-PS C₁₄ 1,2-dipalmitoyl-sn-G-3-PS C₁₆ 1,2-distearoyl-sn-G-3-PS C₁₈ 1,2-dioleoyl-sn-G-3-PS C _{18: 1} (cis) 1,2-dilinoleoyl-sn-G-3-PS C _{18: 2} (cis)

Similarly, mixed chain phosphatidylserines were used manufactured. The starting material was 1-myristoyl-2- Benzyl-sn-glycerol, 1-palmitoyl-2-benzyl-sn-glycerol or 1- Stearoyl-2-benzyl-sn-glycerin used. These substances were described as for 1,2-dibenzyl-sn-glycerol in the used there synthesis path. The following connections were made receive:

1-myristoyl-2-oleoyl-sn-glycero-3-phosphoserine
1-myristoyl-2-linoleoyl-sn-G-3-PS
1-palmitoyl-2-oleoyl-sn-G-3-PS
1-palmitoyl-2-linoleoyl-sn-G-3-PS
1-stearoyl-2-oleoyl-sn-G-3-PS
1-stearoyl-2-linoleoyl-sn-G-3-PS

Claims (12)

1. Use of a chemically defined 1,2-substituted phosphatidylserine of the general formula (I) wherein R ₁ and R _{2 are the} same or different and each represent a saturated or unsaturated, branched or straight-chain hydrocarbon radical having 8 to 24 C-Ato men, or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of diseases in which it to depression and / or disorders of brain function functions, in particular to lessening or loss of memory functions. 2. Use according to claim 1, characterized in that R ₁ and R _{2 are the} same. 3. Use according to claim 1 or 2, characterized, in that -OCO-R₁ and -OCO-R₂ are each selected from the group consisting of of caprylic, capric, lauric, myristic, palmitic, Stearic, oleic, linoleic, cis-11-eicosene, eruca or nerve acid residues are selected. 4. Use according to claim 1 or 2, characterized in that R ₁ and R _{2 represent} mono- or polyunsaturated, straight-chain hydrocarbon radicals. 5. Use according to claim 4, characterized in that R ₁ or / and R _{2 are} oleic acid residues. 6. Use according to claim 5, characterized, the compound of general formula (I) is sodium 1,2-dioleoyl-phosphatidyl- (S) -serine. 7. Medicaments for the treatment of diseases in which there is depression or / and disorders of Hirnlei stungsfunktionen, in particular to lessening or loss of loss of memory functions, characterized in that it has as active ingredient one or more chemically defined th 1,2-substituted Phosphatidylserines of the general formula (I) wherein R ₁ and R _{2 are the} same or different and each represent a saturated or unsaturated, branched or straight-chain hydrocarbon radical having 8 to 24 C-Ato, or a pharmaceutically acceptable salt thereof, optionally together with pharmaceutically customary carrier, auxiliary, filling Contains - and diluents. 8. Medicament according to claim 7, characterized in that R ₁ and R _{2 are the} same. 9. Medicament according to claim 7 or 8, characterized in that R ₁ and R _{2 represent} mono- or polyunsaturated, straight-chain hydrocarbons. 10. Medicament according to claim 7, characterized, that it is active as sodium 1,2-dioleoyl-phosphatidyl Contains (S) -serine. 11. Medicament according to claim 7, characterized, that it as the active ingredient sodium 1,2-dipalmitoyl-phosphati dyl (S) -serine contains. 12. Medicament according to one of claims 7 to 11 for the Therapy of Parkinson's disease, Alzheimer's disease, the Brain organic psychosyndrome, AIDS dementia, depres severe pseudodemes, dementia syndromes, deliria, Intoxications, withdrawal syndromes and / or cytopathic Influences.