EP0112872A1

EP0112872A1 - New glycerine derivatives for the synthesis of phospholipids

Info

Publication number: EP0112872A1
Application number: EP83902086A
Authority: EP
Inventors: Hansjörg EIBL
Original assignee: Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
Current assignee: Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
Priority date: 1982-07-06
Filing date: 1983-07-06
Publication date: 1984-07-11
Also published as: DE3239817A1; JPH041759B2; JPS59501211A; ATE30030T1; WO1984000367A1; DE3373906D1; EP0099068A1; EP0099068B1

Abstract

New derivatives of glycerine having the formulas (I, II and III), wherein the symbol Z represents the methyl rest, a beta-unsaturated alkyl rest, an alkyl-aryl rest or a benzyl rest, or a cation equivalent, wherein the symbols R<1> and R<2>, being similar or different, represent a hydrogen atom, a rest R or an acyl rest -COR or a trityl rest with the proviso that it is in the position 1 or 3, wherein R represents an alkyl rest optionally unsaturated and/or substituted by halogen, hydroxy, carboxy, cyclo-alkyl or aryl, wherein the symbol X represents O, NH or N(R<3>) and the symbol R<3> when the symbol X represents O or NH represents an alkyl rest optionally unsaturated and/or substituted by halogen, amino, methyl-amino, dimethyl-amino, trimethyl-ammonio, hydroxy, carboxy, cyclo-alkyl or aryl, or when X is O, also D- or L-20-tert.-butyl-oxycarbonyl-amino-2-tert.-butyloxycarbonylethyl, 1,2- or 2,3-isopropylidene-dioxypropyl, 1,2-, 1,3 or 2,3-dibenzyloxy-propyl, 1,2,3,4,5-penta-benzyloxy-cyclohexyl or N-alkyl-amino-alkyl, or else when X is NH, also 1,2-dihydroxypropyl and when X is N(R<3>), of similar or different meaning, represent hydrogen or an alkyl rest optionally unsaturated and/or substituted by halogen, cyclo-alkyl or aryl. The compounds of the invention are valuable intermediates and have, at least partially, an anti-tumoral activity.

Description

New glycerin derivatives for the synthesis of phospholipids

The invention relates to new glycerol derivatives for the synthesis of phospholipids, such as diester, monoester, monoether, ether / ester and diether phospholipids and corresponding phosphoric acid amide analogs.

Glycerin phosphatides are extremely important biologically and technically. Isolating pure, uniform glycerol phosphatides from natural tissues is extremely difficult and costly. Their synthesis is particularly problematic when glycerol phosphatides are to be produced in which glycerol with very specific radicals should be acylated in a very specific position.

E. Cubero Robles et al (Rec. Trav. Chim. 86 (1976),

762; Biochim. Biophys. Acta 187 (1969), 520), for example, describe a synthesis of glycerol phosphatides with mixed fatty acid residues by acylation of 1-palmitoyl-sn-glycerol-3-phosphorcholine and fatty acid anhydrides in the presence of Na ₂ O. However, this method is used to a great extent Exchange of the acyl groups takes place, whereby the selectivity, ie very targeted introduction of certain acyl residues, is greatly reduced in a certain position (cf. KM Vf. Keough, PJ Davis, Biochemistry 18 (1979), 1453). Another disadvantage of this method is that during the acy lation step, a migration of the phosphoric acid residue from the 3 to the 2 position takes place to a considerable extent. Rearrangements of both fatty acids and phosphate residues can also occur during the reacylation of monoacylglycerol phosphoric acid esters (cf. A. Plückthun and EA Dennis, Biochemistry (1982), 21, 1743 - 1750; H. Eibl, Chem. Phys. Lipids ( 1980), 26, 405-429; H. Eibl, Liposomes: From Physical Structure to Therapeutic Application: Chapter 2: Phospholipid Synthesis (1981), 19-59; CG Knight, Ed., Elsevier, Amsterdam).

The invention is therefore based on the object to provide starting materials and ways of producing glycerol phosphates (phospholipids) in which the problems mentioned do not occur and which have a defined distribution of acyl, alkyl and phosphate residues over positions 1, 2 and 3 of the Allow glycerol molecule. This object is achieved with the present invention.

The invention relates to glycerol derivatives of For my I, II and III

wherein Z is methyl, optionally a phenyl radical-containing alkyl with a CC multiple bond in the β-position, benzyl or an equivalent of a physiologically compatible cation, preferably methyl, R ¹ and R ^{2 are} identical or different and a hydrogen atom, the radical R or are the acyl radical -COR, in the 1- or 3-position also trityl, R is alkyl having 1 to 24 carbon atoms or halogen, hydroxy or carboxyalkyl having 2 to 24 carbon atoms, where alkyl can be straight-chain or branched and can contain double or triple bonds , and can be substituted by cycloalkyl having 3 to 6 carbon atoms or by aryl radicals, and in which a hydroxyl and / or carboxy group can also contain a protective group, X has the meaning O, NH or N (R ³ ), - and R ³ , if X = 0,

Alkyl having 1 to 20 carbon atoms or halogen,

Hydroxy- or carboxyalkyl or also amino-, methylamino-, dimethylamino- or trimethylammonioalkyl with 2 to 20 carbon atoms, where alkyl can be straight-chain or branched and contain double or triple bonds, and be substituted by cycloalkyl with 3 to 6 carbon atoms or by aryl radicals means, in which a hydroxyl and / or carboxy group can also contain a protective group, D- or L-2-tert-butyloxycarbonylamino-2-tert-butyloxy-carbonylethyl, 1,2- or 2,3-isopropylidene- dioxy-propyl, 1,2-, 1,3-or 2,3-dibenzyloxypropyl, 1,2,3,4,5-pentabenzyloxy-cyclohexyl or aminoalkyl and N-alkylaminoalkyl with 2 up to 14 carbon atoms in the alkyl radicals, in which the amino groups can also contain protective groups, means when X = NH, alkyl with 1 to 20 carbon atoms or halogen, hydroxy or carboxyalkyl or also amino, methylamino, dimethylamino or tri methylammonioalkyl having 2 to 12 carbon atoms, where alkyl can be straight-chain or branched and can contain double or triple bonds, and can be substituted by cycloalkyl having 3 to 6 carbon atoms or by aryl radicals, or 1, 2-dihydroxypropyl, in which a hydroxyl and / or Carboxy group may also contain a protecting group and, if X = N (R ³ ), is different or preferably the same and is hydrogen, alkyl having 1 to 20 carbon atoms or haloalkyl having 1 to 12, preferably 2 to 12 carbon atoms, wherein an alkyl group is straight-chain or branched and can contain double or triple bonds and substituted by cycloalkyl having 3 to 6 carbon atoms or by aryl radicals can be means.

An alkyl group can be branched or, preferably, straight chain and can contain double or triple bonds. An alkyl group with a double bond is, for example, the allyl radical serving as a protective group.

Cycloalkyl is, for example, cyclopropyl, cyclobutyl, cyclopentyl, methylcyclopentyl and preferably cyclohexyl.

An aryl radical is in particular one having 6 to 14 carbon atoms, preferably naphthyl- (1) or - (2), and especially phenyl. An alkyl group substituted with a phenyl radical is, for example, the benzyl group serving as a protective group. An acyl group -COR is derived in particular from saturated and unsaturated natural fatty acids, such as, for example, behenic, lauric, stearic, palmitic, myristic, capric or arachidic acid, and oleic, linoleic or arachidonic acid and more unsaturated fatty acids.

Halogen can be fluorine, bromine or iodine and is especially chlorine.

A physiologically acceptable cation Z is preferably a mono-, di- or trivalent physiologically acceptable cation, such as. As sodium, potassium, calcium and especially lithium. If Z is alkyl with β-unsaturation, it is preferably alkyl or its homologues.

Protecting groups for the hydroxyl group are, for example, benzyl, allyl, trityl, tetrahydropyranyl, mesyl and tosyl; for the carboxy group, for example, tert-butyl.

The precursors of the compounds, of the formulas I, II or III with free hydroxyl groups on the phosphorus, can be obtained analogously to a synthetic route known for glycerol derivatives (cf. for example D. Arnold, HU Weltzien and O. Westphal, Liebigs Ann. Chem. 709 (1967 ), 234 to 239; HU Weltzien and O. Westphal, Liebigs Ann. Chem. 709 (1967), 240 to 243; K. Eibl and 0. Westphal, Liebigs Ann. Chem. 709 (1967), 244 to 247) ; they are preferably known from one of the same filing dates for similar compounds in the same applicant's German patent application P 31 30 867 by the same applicant on August 4, 1981 or in German patent application "New D-mannitol derivatives as starting products for the synthesis of phospholipids" process. The conversion into the methyl ester is then carried out in a manner known per se, for example by reaction with methanol.

The preparation of the glycerol derivatives of the formulas I, II or III, in which Z is an equivalent of a physiologically compatible cation, can in particular be carried out by reacting the corresponding methyl ester (Z = CH ₃ ) with a halide of Z, for. B. with lithium bromide.

It has been shown that starting from the intermediates of the formulas I, II or III, glycerol derivatives with various radicals can be prepared in a simple and economical manner in a position-specific manner and with high selectivity.

The compounds of formula I lead to phospholipids of natural configuration (sn-3-phosphates), those of formula II and III to compounds with non-natural configurations (sn-1- or -2-phosphates).

The glycerol derivatives mentioned above and below can exist both as optically pure stereoisomers and as racemates.

With two same-chain fatty acids or alkyl residues and one phosphate residue, three stereoisomers are possible; in the case of two different fatty acids or alkyl residues and one phosphate residue, six stereoisomers are possible, all of which can be obtained from the intermediate compounds I, II and III.

The methyl group in formulas I, II and III is superior to corresponding benzyl groups in many respects. It has sufficient stability for the necessary ones Reaction steps, for example the phosphoric acid methyl esters of the formulas I, II and III are stable under catalytic hydrogenolysis, acidic trityl and propenyl elimination under the selected conditions. With lithium bromide, the methyl group can be removed easily and without destroying the phospholipid molecule.

The invention therefore also relates to a process for the further processing of the compounds of the formulas I, II or III for the preparation of phospholipids, which is characterized in that

a) for the preparation of diester phospholipids with the same acyl residues, a compound in which R ¹ and R ² = hydrogen, acylated and then demethylated with lithium bromide, or b) for the preparation of diester phospholipids with different acyl residues from a starting compound goes out in which R ¹ and R ^{2 are} different and R ¹ alkanoyl, benzyl, allyl or trityl and R ² alkanoyl,

Is benzyl or allyl, where a radical R ¹ or R ^{2 is} an alkanoyl group, splits off a benzyl, allyl or trityl group R ¹ or R ² , acylates the released hydroxyl group and then demethylates it with lithium bromide, or c) to produce monoester phospholipids starts from a starting compound in which R ¹ and R ^{2 are} different and R ^{1 is} alkanoyl, benzyl, allyl or trityl, R ² alkanoyl, benzyl or allyl, where a radical R ¹ or R ^{2 is} an alkanoyl group, and with

Lithium bromide demethylated, or d) for the preparation of mono-ether phospholipids starts from a compound in which R ¹ and R ^{2 are} different and R ^{1 is} alkyl, benzyl, allyl or trityl, R ^{2 is} alkyl, benzyl or allyl, one radical R ¹ or R ^{2 is} an alkyl group, and demethylated with lithium bromide, or e) for the preparation of ether / ester phospholipids, starts from a compound in which R ¹ and R ^{2 are} different and R ^{1 is} alkyl, benzyl, allyl or trityl be means R ^{2 is} alkyl, benzyl or allyl, where a radical R ¹ or R ^{2 is} an alkyl group, splits off a benzyl, allyl or trityl group R ¹ or R ^2, acylates the resulting hydroxyl group, and then demethylates with lithium bromide, or f) for the preparation of diether phospholipids from one

Compound in which R ¹ and R ^{2 are} alkyl and demethylated with lithium bromide,

and if necessary or desired, in a compound obtained according to a) to f) cleaves off protective groups which are known per se and / or aminates the phosphorus-containing radical or converts it to another phosphorus-containing radical, the cleavage of benzyl, allyl or trityl can also be carried out after the amination or conversion into another phosphorus-containing radical. These processes also make it possible to convert a compound of the formula I, II or III into another compound of the formula I, II or III with other radicals R ¹ , R ² , R ³ or Z.

If desired or expedient, the processes can also be carried out by starting from a product obtained by one of the process steps and carrying out the remaining process steps. Taking into account all residues present in the molecule, it is also possible, for example, to interchange the order of the individual process steps.

The acylation of a free hydroxy group can be carried out by Reaction with acid chlorides in the presence of triethylamine or pyridine can be carried out (cf. H. Eibl and 0. Westphal, Liebigs Ann. Chem. 709 (1967), 244). However, it is also possible to work according to other acylation methods known per se (for example as described by Gupta et al, Proc. Nat. Acad. Sci. USA 74 (1977), 4315). Acy lation is particularly easy with free fatty acid and dicyclohexylcarbodiimide in the presence of dimethylaminopyridine.

The trityl group is cleaved off under weakly acidic conditions, preferably at a pH of 4 to 6, it being easy to determine the most favorable value taking into account the other substituents in the molecule. The allyl and benzyl protective groups are completely stable. The reaction can be carried out in an aqueous or aqueous-organic medium, but also in a purely organic medium, for example in absolute ethanol, in the presence of HCl or H ₂ SO ₄ . The organic solvent can be a

Water-miscible or an inert solvent that is only partially or only slightly miscible with water. The reaction, particularly when working in a two-phase system, advantageously takes place with vigorous stirring. The temperature is generally 20 to 80 ° C. To improve the solubility, it may be expedient to add a higher alcohol, such as propanol (2), in a small amount.

The benzyl group is split off by catalytic hydrogenolysis. The reaction conditions correspond to the usual conditions. In particular, the hydrogenolysis is carried out in an inert solvent, such as ethanol, in the presence of a palladium or platinum / palladium catalyst, preferably at room temperature and under normal pressure (cf. H. Eibl et al, Liebigs Ann. Chemie, 738 (1970), 161).

The cleavage of the allyl group (rearrangement into propenyl and subsequent cleavage of propenyl) can be carried out by two different methods, namely 1) under alkaline conditions, such as with potassium tert-butoxide in dimethylformamide and subsequent cleavage with bromine in buffered solution at a pH - Value around 5 to 6, or 2) by rearrangement in the presence of a palladium (carbon) catalyst to form the propenyl group which spontaneously cleaves under these conditions, expediently in 80% methanol, which in the aqueous phase is 20% Contains formic acid, is carried out at reflux temperature. In general, variant 1, i.e. H. cleavage with bromine is preferred. Iodine can also be used to split off the propenyl group in the 1 position (Eibl and Lands, Biochemistry 9 (1970), 423). However, while the propenyl group cannot be split off in the 2-position with iodine, such a split-off can surprisingly be carried out completely and in a few minutes using bromine.

The methyl phosphoric acid is demethylated with lithium bromide by boiling in a suitable organic solvent, preferably in methyl ethyl ketone.

The amination of the residue containing phosphorus can be carried out in a manner known per se (cf., for example, BH Eibl and A. Nicksch, Chem. Phys. Lipids, 22 (1978), 1; W. Diembeck and H. Eibl, Chem. Phys Lipids, 24 (1979), 237), as well as the subsequent alkylation of a free amino group. Compounds of the formula I and II according to the invention are also of great importance because of their particular effectiveness in the growth of tumors. It has been found that, in particular, the glycerol derivatives of the general formula I and II, in which R ¹ denotes the radical - (CH ₂ ) _m -CH ₃ , where m is 13-19, R ^{2 is} a hydrogen atom and, if X = O, R ^{3 is} alkyl with 1 to 12 carbon atoms, alkyl with 2 to 11 carbon atoms substituted by hydroxy or trityloxy, alkyl substituted with carboxy or tert-butyloxycarbonyl to 1 to 11 carbon atoms, 2- (amino- or tert-butyloxycarbonylamino) -2-tert-butyloxycarbonylethyl, 1,2-dihydroxy- or 1,2-isopropylidenedioxy-propyl, pentahydroxy- or pentabenzyloxy-cyclohexyl, or by bromine, amino, methylamino, dimethylamino, trimethylammonio, tert.- Butyloxycarbonylamino, tert-butyloxycarbonyl-methylamino W-substituted alkyl with 3-11 carbon atoms means, when X = NH, R ^{3 is} alkyl with 1 to

12 carbon atoms, alkyl substituted by bromine, hydroxyl, trityloxy, amino, methylamino, dimethylamino or trimethylammonio ധ- alkyl having 2 to 11 carbon atoms, alkyl substituted by carboxy or tert-butyloxycarbonyl having 1 to 11 carbon atoms or 1,2-dihydroxy- or 1,2-isopropylidenedioxy-propyl, and, when X = N (R ³ ), R ^{3 is} different or preferably the same and is alkyl with 2 to 4 carbon atoms or W-chloroalkyl with 1 to 4 carbon atoms, a pronounced tumor growth inhibitor Have an effect. This effect is shown by both the optically pure compounds and the Raemates. A particularly good effect is shown by compounds of the formula II and in particular I in which R ^{1 is} n-octadecyl, R ^{2 is} hydrogen, X = 0 and R ^{3 is} 3-trimethylammoniopropyl.

The pronounced tumor growth-inhibiting effect of the Lyso compounds (1-alkyl-2-OH) according to the invention are based in part on a completely new active principle. They are not detoxified by acyltransferases or only very slowly. This is based on some of the lyso compounds according to the invention in which the radical R ³ z. B. one by

Amino or alkyl-substituted amino ധ-substituted alkyl group with more than 3 carbon atoms, insisting that between P and N there are at least 3 methylene groups, such as. B. in the grouping

In such a case, acyltransferases no longer attack; Phosphorus lipases A _{2 are} still attacking, but the acyltransferases can no longer reactylate.

The compounds of the formulas I and II, in which R ^{1 is} the

Residue - (CH ₂ ) _m -CH ₃ , in which m is 13 to 19 and R ² denotes an acyl residue COR, act as a precursor of the above-mentioned tumor-active substances in natural form. With phospholipase A ₂ they go into the active substances in vivo.

In contrast to the 1-alkyl-2-OH compounds into which they are first converted enzymatically, the 1-alkyl-2-acylglycerol derivatives are not hemolytic; they can therefore be dosed higher without any problems. It is advantageous to use matrix molecules with stable ether structures that are not cleaved by phospholipase A ₂ , such as in particular, for. B. 1-octadecyl-2-oleyl compounds.

The effectiveness of compounds on the growth of tumors is expedient on tumors in experimental animals proven. Various experimental tumors are used for this, for example the Ehrlich-Ascites tumor, a methylcholanthrene-induced tumor and a myeloma tumor in mice, and also a chemically induced rat tumor. The anti-tumor substances are administered parenterally into the tumor-bearing test animals. Intravenous and intra- or subcutaneous administration is preferred. Oral applicability is with a correspondingly higher dosage of the anti-tumor agent and with a physiologically compatible preparation, for. B. in capsules, not excluded.

The invention therefore also relates to medicaments which contain one or more of the abovementioned tumor growth-inhibiting compounds of the formula I or II as active ingredient. In addition to the customary pharmaceutical formulations and / or diluents, these medicaments may also contain, in addition to the compounds of the formulas I and II, to assist therapy, if appropriate, provided that these, together with the compounds according to the invention, do not show any undesirable side effects. The dosage and application essentially correspond to the conditions given for the anti-tumor agents of DE-OS 2 619 686, but due to the higher effectiveness, lower doses and / or less frequent administration can also be considered. In the case of parenteral administration, it has proven expedient to use about 0.05 to 5 mg / kg of body weight as the dosage. In order to allow the anti-tumor agents to persist in the circulation over a long period of time, it is often sensible to apply the agents daily or at 2 to 3 day intervals. One which has proven to be a particularly favorable pharmaceutical composition is one in which the active ingredient component according to the invention consists of a mixture of about 80-95% by weight of 1-alkyl-2-acyl compound and about 20-5% by weight of 1-alkyl -2-OH connection exists. Such a composition is not hemolytic due to the higher content of 1-alkyl-2-anyl compound.

Schemes A, B and C below show the possibilities of variation in the radical containing phosphorus (polar region) without restricting it (A: phospholipids; B and C: phosphatidic acid amides). Starting from the triphosphonic acid esters (A) or the corresponding phosphoric acid amides (B, C), all essential phospholipids of natural membranes or their analogues can be prepared in a simple manner.

The known disadvantages of the synthesis of glycerol phosphatides, in particular the migration of the acyl residues and / or of the

Phosphoric acid residue, avoided. It has been shown that the compounds of the formulas I, II and III are very good starting materials for the preparation of glycerol phosphatides, it being possible for each individual process step to be carried out under conditions which do not affect the other substituents.

In summary, it can thus be stated that the process according to the invention is a very selective, simple and economical method for the production of phospholipids, which is particularly important for the stereo- and position-specific preparation of glycerol phosphatides with two or three different radicals. The following examples illustrate the invention without restricting it.

Scheme A;

Diester, monoester, ether / ester, monoether and diethylphospholipids (R = basic glycerol with residues R ₁ and R ₂ ; X = O).

Scheme B:

Diester, monoester, ether / ester, monoether and diethylglycerol phosphoric acid amides (R = glycerol base with the residues R ¹ and R ² ; X = NH).

Scheme C:

Diester, monoester, ether / ester, monoether and diethylglycerol-phosphoric acid dialkylamides (R = glycerol base with the residues R ¹ and R ² ; X = N (R ³ )).

Examples

Diester phospholipids of the same chain

a) 1,2-dibenzyl-sn-glycerol

The starting product is 3,4-isopropylidene-D-mannitol (CF Wiggins, J. Chem. Soc. 13, 1946). A solution of 0.1 mol in 1 l of toluene is mixed with 0.5 mol of K-tert-butoxide and boiled under reflux. 0.5 mol of benzyl chloride is added dropwise with stirring. After completion of the reaction (TLC control), the reaction mixture is extracted with water and the toluene phase is spun in. The residue is taken up in 1 l of methanol / 2-propanol 1: 1 (v / v) for deacetonation, 50 ml of 2N-H ₂ SO _{4 are} added and the mixture is boiled under reflux. When the reaction is complete (TLC control), 1 l of diisopropyl ether is added and the mixture is washed twice with 1 l of water. The diisopropyl ether phase is evaporated and the intermediate, 1, 2,5,6-tetrabenzyl-D-mannitol is purified by chromatography on silica gel. The yields are 90-95%.

1,2,5,6-tetrabenzyl-D-mannitol (C ₃₄ H ₃₈ O ₆ ; 542.68) calc. C 75.25 H 7.06 found. C 75.11 H 6.99

1, 2,5,6-tetrabenzyl-D-mannitol (0.1 mol) is dissolved in 500 ml of benzene, and lead tetraacetate (approx. 0.1 mol) is added in portions until the starting product is completely converted (TLC control) . It is washed twice with 500 ml of water each time, the benzene phase is rotated in and the residue is taken up in 500 ml of methanol. The solution of the aldehydes in methanol is added in portions NaBH ₄ (approx. 0.1 mol) was added. After the reaction has ended, 500 ml of diisopropyl ether are added and the mixture is washed with water. The diisopropyl ether phase is spun in and the residue is chromatographed on silica gel. The yield of 1,2-dibenzyl-sn-glycerol is 95%; Rotation value (in substance).

1,2-dibenzyl-sn-glycerol (C ₁₄ H ₂₀ O ₃ ; 272.35) calcd. C 74.97 H 7.40 found. C 74.91 H 7.40.

b) phosphorylation

1, 2-Dibenzyl-sn-glycerol (0.1 mol) are dissolved in 500 ml THF and triethylamine (0.2 mol) is added. The solution is added dropwise with vigorous stirring at 15 ° C. to phosphorus oxychloride (0.15 mol) in 100 ml of THF. After the reaction has ended (TLC control), the precipitated triethylamine hydrochloride is filtered off, 100 ml of toluene are added and the mixture is concentrated in vacuo at 40 ° C. (removal of excess POCl ₃ ). The oily residue consists mainly of 1,2-dibenzyl-sn-glycero-3-phosphoric acid dichloride, which is dissolved in 500 ml of THF and added dropwise with triethylamine (0.3 mol) and then with the corresponding alcohols, e.g. B. N-BOC-serine tert-butyl ester (0.15 mol) (BOC = butyloxycarbonyl) is added. After the reaction is complete, methanol (1 mol) is added and the mixture is worked up after the reaction is complete. 500 ml of diisopropyl ether are added and the organic phase is extracted with water. The diisopropyl ether phase is spun in and the residue is chromatographed on silica gel. 1,2-Dibenzylsn-glycero-3-phosphomethyl-N-BOC-serine tert-butyl ester is obtained in yields of 80%. 1,2-dibenzyl-sn-glycero-3-phospho-methyl-N-BOC-serine tert-butyl ester

(C ₃₀ H ₄₄ NO ₁₀ P; 609, 67) calcd. C 59, 1 0 H 7, 28 N 2, 30 P 5, 08 found. C 58, 95 H 7, 31 N 2, 1 7 P 5, 01.

c) Katalyti see debenzylation

1,2-Dibenzyl-sn-glycero-3-phosphomethyl-N-BOC-serine tert-butyl ester (0.1 mol) are dissolved in 400 ml THF and with 10 g Pd / C (10% Pd on activated carbon ) and 1 g of palladium black. The mixture is left under stirring under a H ₂ atmosphere until the H ₂ uptake is complete. The reaction is quantitative. The catalyst is filtered off and the solvent is removed in vacuo. The oily residue is directly implemented further.

sn-glycero-3-phosphomethyl-N-BOC-serine tert-butyl ester (C ₁₆ H ₃₂ NO ₁₀ P; 429.415) calcd. C 44.75 H 7.51 N 3.26 P 7.21 found . C 44.59 H 7.39 N 3.14 P 7.12.

By appropriate reaction with other alcohols, further central intermediates can be obtained, for example:

sn-glycero-3-phosphomethyl-bromoethyl ester (C6 H ₁₄ Br O ₆ P; 269.07) calcd. C 26.78 H 5.25 P 11.51 found. C 26.63 H 5.21 P 11.01.

d) acylation

The intermediates with two free hydroxyl groups in the sn-1 and sn-2 positions of the glycerol can now be used with the desired fatty acids are implemented, for example with palmitic acid or with oleic acid. For this purpose, 0.1 mol are dissolved in 300 ml of methylene chloride, the fatty acids (0.25 mol) in 200 ml of methylene chloride, with dicyclohexylcarbodiimide (0.25 mol) and 4-dimethylaminopyridine (0.05 mol) are added. After the reaction has ended, the precipitated urea is filtered off with stirring at 20 ° C., washed with water and the methylene chloride phase is spun in. The residue is purified on silica gel by chromatography. The yield of acylated product is 95%.

1, 2-Dioleoyl-sn-glycero-3-phospho-methyl-N-BOC-L-Serintert.-butyl ester (C ₅₂ H ₉₆ NO ₁₀ P; 926.32) calcd. C 65.17 H 10.10 N 1.46 P 3.23 found C 64.86 H 10.11 N 1.47 P 3.17.

e) Demethylation of the phosphoric acid diester with the protected serine

(0.1 mol) is mixed with a solution of LiBr (0.5 mol) in ethyl methyl ketone, 500 ml, and boiled under reflux for 60 min. The split is quantitative. Diisopropyl ether is added and the mixture is extracted with water. The diisopropyl ether phase is spun in and the residue is directly reacted further.

f) cleavage of the serine protective groups

The residue from e) (0.1 mol) is dissolved in 300 ml of methylene chloride and 150 ml of trifluoroacetic acid are added at 0 ° C. After adding 30 ml of 70% HClO ₄ , the mixture is stirred at 0 ° C. for 60 minutes. The split is complete. 300 ml of chloroform, 600 ml of water and 600 ml of methanol are added. The chloroform phase is added to 600 ml of 1M NaHCO ₃ . After adding 600 ml of CH ₃ OH shaken vigorously and the chloroform phase is spun in after phase separation. The residue is chromatographed on silica gel. The yield is 95%, based on the phosphoric triester.

1,2-dioleoyl-sn-glycero-3-phosphoserine

(C ₄₂ H ₇₇ N Na O ₁₀ P; 810.05) calcd. C 62.28 H 9.58 N 1.73 P 3.82 found. C 62.09 H 9.41 N 1.49 P 3.69.

mixed chain a) phosphorylation

1-Trityl-2-benzyl-sn-glycerol (0.1 mol) is dissolved in 500 ml THF and triethylamine (0.2 mol) is added. The solution is added dropwise at 15 ° C. with vigorous stirring to phosphorus oxychloride (0.15 mol) in 100 ml of THF. After the reaction has ended, the mixture is filtered, 100 ml of toluene are added and the mixture is concentrated in vacuo at 40 ° C. (excess triethylamine and phosphorus oxychloride are drawn off). The residue, mainly 1-trityl-2-benzyl-sn-glycero-3-phosphodichloride, is taken up in 500 ml of THF and triethylamine (0.3 mol) is added. For example, bromoalkanols (0.2 mol) are added dropwise to this solution at 30 ° C. and, after the reaction has ended, methanol (1 mol) is added. The methanolysis is quickly completed at 30 ° C (approx. 2 hours). 500 ml of diisopropyl ether are added and the organic phase is extracted with water. The diisopropyl ether phase is spun in.

b) Detritylation

The oily residue (0.1 mol) from a) is concentrated in 500 ml CH ₃ OH, the 5 ml. Contain H ₂ SO ₄ , taken up and heated to 50 ° C. The reaction is complete after 10 minutes, the mixture is cooled to 0 ° C. and the crystal is filtered linen trityl methyl ether. After the addition of chloroform, 500 ml, the mixture is washed with 500 ml of 1M NaHCO ₃ (pH of the aqueous phase should be at least 5). The chloroform phase is dried over Na ₂ SO ₄ and evaporated.

c) First acylation

The residue from a) is taken up in 500 ml of methylene chloride and mixed with palmitic acid (0.25 mol). After addition of dicyclohexylcarbodiimide (0.25 mol) and 4-dimethylaminopyridine (0.05 mol), the mixture is stirred at 20 ° C. for 3 hours. The precipitated urea is filtered off, the filtrate is washed with water and the methylene chloride is removed in vacuo. The residue is chromatographed on silica gel. The yield is 75% based on 1-trityl-2-benzyl-sn-glycerol.

d) Catalytic debenzylation

1-Palmitoyl-2-benzyl-sn-glycero-3-phospho-methyl-bromo-alkyl ester (0.1 mol) is dissolved in diisopropyl ether, 500 ml, and with 10 g Pd / C catalyst (10% Pd) and

1 g of palladium black added. The mixture is stirred under an H ₂ atmosphere until the hydrogen uptake is complete. 1-Palmitoyl-sn-glycero-3-phosphomethyl-bromoalkyl ester is obtained quantitatively. The catalyst is filtered off, THF is removed from the filtrate and the residue is taken up in 500 ml of carbon tetrachloride.

e) Second acylation

The carbon tetrachloride solution of the phosphoric acid triester (0.1 mol) is mixed with dicyclohexylcarbodiimide (0.25 mol) and oleic acid (0.25 mol). After adding 4-dimethyl-aminopyridine, the mixture is stirred at 20 ° C. until the reaction is complete (TLC control).

The precipitate is separated off by filtration, ex tracts with water and concentrates the carbon tetrachloride phase in vacuo. The residue is chromatographed on silica gel. 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphomethyl-bromoalkyl ester is obtained in yields of approx. 95%.

f) Demethylation The phosphoric acid triester (0.1 mol) from e) is mixed with 0.5 mol of LiBr in 500 ml of ethyl methyl ketone and boiled under reflux for 60 minutes. The yield of Li salt of 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-bromoalkyl ester is quantitative.

g) Amination The amination of the bromoalkyl esters can be carried out in a manner known per se (cf., for example, BH Eibl and A. Nicksch, Chem. Phys. Lipids 22 (1978) 1; W. Diembeck and H. Eibl, Chem. Phys. Lipids 24 (1979) 237). The reaction with the amine, e.g. B. trimethylamine, z. B. in chloroform / propanol (2) (1/1) as a solvent.

h) Direct amination of the phosphoric acid triesters

The phosphoric acid triesters (0.1 mol) from e) are dissolved in 500 ml of chloroform and mixed with 500 ml of 2-propanol which contains trimethylamine (1 mol). After 24 hours at 50 ° C the reaction is complete. The solvent is removed and 500 ml of chloroform, methanol and water are added. The chloroform phase contains 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-N, N, N-trimethylalkanolamine ester. The solvent is removed and the mixture is purified by chromatography. The yields are more than 90%. Monoester phospholipids a) Demethylation

The products of the first acylation (mixed chain, reaction c)) are demethylated with LiBr; for example, 1-palmitoyl-2-benzyl-sn-glycero-3-phosphomethyl-bromoalkyl ester (0.1 mol) is boiled under reflux with LiBr (0.5 mol) in 500 ml of ethyl methyl ketone. After 60 minutes, the implementation is quantitative. The amination can be carried out as described above under g).

b) Catalytic debenzylation

1-Palmitoyl-2-benzyl-sn-glycero-3-phospho-N, N, N-trimethylalkanolamine ester (0.1 mol) are dissolved in alcohol, 1 l, and with 10 g Pd / C catalyst (10% palladium on activated carbon) and 1 g of palladium black. The mixture is stirred until the hydrogen uptake has ended. Debenzylation is quantitative. The catalyst is filtered off and the solvent is removed in vacuo. The residue is taken up in 100 ml of CHCl ₃ and precipitated with 400 ml of acetone. The precipitate is filtered off and dried.

Mono-ether phospholipids The mono-ether phospholipids can be prepared in exactly the same way as the mono-ester phospholipids. In this case, 1-octadecyl-2-benzyl-sn-glycerol, for example, is used as the starting product.

Ether / ester phospholipids

Ether / ester phospholipids are obtained if, for example, 1-octadecyl-2-benzyl-sn-glycerol is used as the starting product and the procedure for diester phospholipids, mixed-chain, is used. After phosphorylation a), however, the catalytic debenzylation d) and the second acylation e) take place immediately. Then proceed as described there.

Diether phospholipids

1, 2-diether-sn-glycerols, like the 1,2-diester-sn-glycerols, can be converted into the corresponding phospholipids if the same chain procedure is used as for diester phospholipids and process steps b), phosphorylation, e), Demethylation and f), splitting off the protective groups, applies.

Claims

1. Glycerol derivatives of the formulas I, II and III

in which Z is methyl, optionally an alkyl containing a phenyl radical with a CC multiple bond in the β-position, benzyl or an equivalent of a physiologically compatible cation, R ¹ and R ^{2 are} identical or different and are a hydrogen atom, the radical R or the acyl radical - COR means, in the 1- or 3-position also trityl, R is alkyl with 1 to 24 carbon atoms or halogen, hydroxy or carboxyalkyl with 2 to 24 carbon atoms, where alkyl can be straight-chain or branched and contain double or triple bonds, and through Cycloalkyl with 3 to 6 carbon atoms or can be substituted by aryl radicals, and in which a hydroxyl and / or carboxy group also can contain a protective group, X has the meaning 0,

Has NH or N (R ³ ) and R ³ when X = O,

Alkyl having 1 to 20 carbon atoms or halogen, amino, methylamino, dimethylamino, trimethylammonio, hydroxy or carboxyalkyl having 2 to 20 carbon atoms, where alkyl can be straight-chain or branched and can contain double or triple bonds, and by cycloalkyl having 3 to 6 Carbon atoms or can be substituted by aryl radicals means in which an amino, hydroxyl and / or carboxy group can also contain a protective group, or D- or L-2-tert-butyloxycarbonylamino-2-tert-butyloxycarbony1-ethy1-, 1,2- or 2,3-isopropylidenedioxy-propyl-, 1,2-, 1,3- or 2,3-dibenzyloxy-propyl-, 1,2,3,4,5-pentabenzyloxy-cyclohexyl- or N- Alkylaminoalkyl with 2 to 14 carbon atoms in the alkyl radicals, in which the amino groups can also contain protective groups, means, when X = NH, alkyl with 1 to 20 carbon atoms or halogen, amino, methylamino, dimethylamino, trimethylammonio, Hydroxy- or carboxyalkyl with 2 to 12

Carbon atoms, where alkyl can be straight-chain or branched and can contain double or triple bonds and can be substituted by cycloalkyl having 3 to 6 carbon atoms or by aryl radicals, or 1,2-dihydroxypropyl, in which an amino, hydroxyl and / or carboxy group also may contain a protecting group and, when X = N (R ³ ), is the same or different and denotes hydrogen, alkyl having 1 to 20 carbon atoms or haloalkyl having 1 to 12 carbon atoms, in which an alkyl group is straight-chain or branched and double or triple bonds may contain, and may be substituted by cycloalkyl having 3 to 6 carbon atoms or by aryl radicals.

2. Glycerol derivatives of formula I or II according to An Proof 1, characterized in that R ^{1 is} the radical - (CH ₂ ) _m -CH ₃ , where m is 13-19, and R ^{2 is} an acyl radical COR.

3. glycerol derivatives of the formula I or II according to claim 1, characterized in that R ^{1 is} the radical - (CH ₂ ) -CH ₃ , wherein m is 13-19, R ^{2 is} a

Hydrogen atom and, if X = 0, R ³ alkyl with 1 to 12 carbon atoms, alkyl with 2 to 11 carbon atoms substituted by hydroxy or trityloxy, alkyl with 1 to 11 carbon atoms substituted by carboxy or tert-butyloxycarbonyl, 2 - (Amino or tert-butyloxycarbonylamino) -2-tert-butyloxycarbonyl-ethyl, 1, 2-dihydroxy- or 1,2-isopropylidene-dioxy-propyl, pentahydroxy- or pentabenzyloxy-cyclohexyl, or by bromine, amino, methylamino, Dimethylamino, trimethylammonio, tert.-butyloxycarbonylamino, tert.-butyloxycarbonylmethylamino ധ- substituted alkyl with 3 - 11 carbon atoms means, when X = NH, R ^{3 is} alkyl with 1 to 12 carbon atoms, by bromine, hydroxy, trityloxy, amino, methylamino, Dimethylamino or trimethylammonio ധ- substituted alkyl with 2 - 11 carbon atoms, by carboxy or tert-butyloxycarbonyl ധ- subst. Means alkyl having 1 to 11 carbon atoms or 1,2-dihydroxy- or 1,2-isopropylidenedioxy-propyl, and, if X = N (R ³ ), R ^{3 is} different or identical and

Alkyl with 2 to 4 carbon atoms or ധ-chloroalkyl with 1 to 4 carbon atoms.

4. A process for further processing of the compounds of the formulas I, II or III according to one of claims 1 to 3 for the preparation of phospholipids, characterized in that a) for the preparation of diester phospholipids with the same acyl radicals starts from a compound in which

R ¹ and R ² = hydrogen, acylated and then demethylated with lithium bromide, or

b) for the preparation of diester phospholipids with unequal acyl radicals, starting from a starting compound in which R ¹ and R ^{2 are} different and R ¹

Alkanoyl, benzyl, allyl or trityl and R ^{2 is} alkanoyl, benzyl or allyl, where a radical R ¹ or R ^{2 is} an alkanoyl group, splits off a benzyl, allyl or trityl group R ¹ or R ² , acylates the released hydroxyl group and then demethylated with lithium bromide, or

c) for the preparation of mono-ester phospholipids starting from a starting compound in which R ¹ and R ^{2 are} different and R ^{1 is} alkanoyl, benzyl, allyl or trityl, R ² alkanoyl, benzyl or Al lyl, with a radical R ¹ or R ^{2 is} an alkanoyl group and demethylated with lithium bromide, or

d) for the preparation of mono-ether phospholipids starts from a compound in which R ¹ and R ^{2 are} different and R ^{1 is} alkyl, benzyl, allyl or trityl, R ^{2 is} alkyl, benzyl or allyl, with a radical R ¹ or R ^{2 is} an alkyl group and demethylated with lithium bromide, or

e) for the preparation of ether / ester phospholipids starts from a compound in which R ¹ and R ^{2 are} different and R ^{1 is} alkyl, benzyl, allyl or trityl, R ^{2 is} alkyl, benzyl or allyl, with a radical R ¹ or R ^{2 is} an alkyl group, splits off a benzyl, allyl or trityl group R ¹ or R ² which resulting hydroxyl group is acylated and then demethylated with lithium bromide, or

f) for the preparation of diether phospholipids, starts from a compound in which R ¹ and R ^{2 are} alkyl and demethylates with lithium bromide,

and, if necessary or desired, in a compound obtained according to a) to f) splits off protective groups present in a manner known per se and / or the

Amine containing phosphorus aminated or converted into another phosphorus-containing residue, wherein the cleavage of benzyl, allyl or trityl can also take place after the amination or conversion into another residue containing phosphorus.

5. Medicament containing one or more compounds of the formula I or II according to claim 2 or 3.

6. Medicament according to claim 5, characterized in that it contains a mixture of approximately 80-95% by weight of a 1-alkyl-2-acyl compound as claimed in claim 2 and approximately 20-5% by weight of a 1- Alkyl-2-OH compound according to claim 3 included.