DD288826A5 - METHOD FOR PRODUCING NEW, EFFICIENT ANTAGONISTS OF PLAETTE-ACTIVATING FACTOR (PAF) WITH PHOSPHOLIPID STRUCTURE - Google Patents

Abstract

The invention relates to a method for producing novel, effective antagonists of the platelet-activating factor with phospholipid structure. The aim of the invention is to produce by a simple to implement method new PAF antagonists with high activity, good water solubility and plasma stability. The goal is achieved by reacting haloesters of phospholipids with various alicyclic tertiary bases. The thus altered head group essentially determines the PAF-antagonistic action of the compounds. The substances produced according to the invention can be used for the pharmacological control of PAF-mediated reactions and as active ingredients of antiphlogistics, antiallergic agents, antiasthmatics and antithrombotics. {Synthetic phospholipids; PAF antagonists; anti-inflammatory drugs; antiallergics; antiasthmatics; antithrombotics}

Description

where R ^{3 is} an unsubstituted alkyl or alkylene radical having 1 to 3 carbon atoms and 7 is a methylene or methine group and also oxygen or nitrogen which may also be linked to compounds of general formula III by R ³ ,

CH ₀ -OR ¹ CH - R ²

O O ^ CH

wherein R ¹ , R ² and R ³ and η and Z have the meaning given above, are reacted.

2. The method according to claim 1, characterized in that the reactions with a 1.5 to 2 molar excess of II and in a temperature range of 50-6O ⁰ C are performed.

3. The method according to claim 1, characterized in that the reactions using anhydrous halogenated hydrocarbons, such as methylene chloride, chloroform, carbon tetrachloride or anhydrous solvent mixtures of chloroform / dimethylformamide or chloroform / acetonitrile (1: 1) are carried out with exclusion of moisture.

Application of the invention

The invention relates to a process for the preparation of novel phospholipids with PAF-antagonistic properties. PAF is a naturally occurring phospholipid which is a mediator of allergic and inflammatory reactions of pathophysiological importance. Accordingly, the compounds according to the invention can be used for the pharmacological control of PAF-mediated reactions and as active ingredients of anti-inflammatory drugs, antiallergic drugs, anti-asthmatics and antithrombotics.

Characteristic of the known technical solutions

Since the structural elucidation of PAF in 1979 and its characterization as a physiological mediator of allergic and inflammatory reactions, PAF antagonists are increasingly gaining interest as potential agents and drugs. Among the currently known antagonists are: structural analogs of PAF, some naturally occurring lignans, terpenoids and gliotoxins, and a number of synthetic heterocycles (see also: D. HOSFORD, JM MENCIA-HUERTA, C. PAGE, P. BRAQUET: Phytotherapy Research: Vol 1,1,1988.

TVSHEN. S.-B.HWANG, TWDOEBBERJ.C.ROBINS (p.153-1901 and P.BRAQUF.T, JJ ₁ GODFROID [S. 191-2351 in: Platelet-Activating Factor and Related Lipid Mediators; F.SNYDER (Ed .], 1987.

HKMANGOLD (page 1) and J.-J.GODFROID.F.HEYMANSIS ^ B] in: Progress In Biochemical Pharmacology, Vol.22, Biologically Active Ether Lipids (P.BRAQUEI, HK MANGOLD, BB VARGARIG [Eds.] , 1988.

M.N. CHANG: Drugs Future 11,869,1987).

Many of the natural products described and synthetic heterocycles with PAF-antagonistic action are difficult to dissolve in water and therefore can not be used for pharmacological purposes. On the other hand, the isolation and synthesis usually requires a high experimental effort, which meets the requirements of a technical process only inadequately. The described Phospholipidabkömmlinge with PAF-antagonistic action are generally poorly soluble in water. In addition, they are partly abgobaut in the plasma. Their preparation is usually characterized by very low yields.

Object of the invention

The aim of the invention is to develop a technically simple to implement process for the preparation of new PAF antagonists with phospholipid structure, which are characterized by high activity, good water solubility and plasma stability.

Explanation of the essence of the invention

The object of the invention is to find an easy-to-implement synthetic route for the preparation of new, effective PAF antagonists. Ale Auegangsverbindungen are used in eich known catfish halophosphorus of phospholipids of general formula I.

CH ₀ -OR ¹ I

ο ate

wherein R ^{1 represents} a long-chain, unsubstituted alkyl or alkylcarbamoyl radical having from 16 to 20 carbon atoms, R ^{2 represents} a methoxy, ethoxy, propoxy or benzoxy group or a methyl, ethyl or n-propyl to n-pentyl radical or a hydroxy group η are the numbers 4 to 10 and X is chlorine or bromine, with acyclic tertiary bases of the general formula II,

where R ^{3 is} an unsubstituted alkyl or alkylene radical having 1 to 3 carbon atoms and Z is a methylene or methine group and also oxygen or nitrogen which may also be linked to R ³ to give compounds of the general formula III,

CH ₀ -OR ¹

CH-R ^

(III)

// O

wherein R ¹ , R 'and R ³ and η and Z have the meaning given above, are reacted. The reactions are carried out in dried halohydrogen stages such as methylene chloride, chloroform or carbon tetrachloride or in dried solvent mixtures of chloroform / dimethylformamide and chloroform / acetonitrile (1: 1) with exclusion of moisture. To increase the yield, the amount of the base II is preferably used in a 1.5 to 2molaren excess. The reactions are performed in a closed vessel in the temperature range of 50 is 6O ⁰ C. Surprisingly, the compounds of the invention show extremely favorable pharmacological properties. They are characterized by good water solubility and high plasma stability and inhibit even in low concentrations PAF-mediated reactions. These results were not expected in the current state of the art and represent a valuable addition to the drugs known in the art.

The invention will be explained on Ausführungsboispiolon. example 1

1 -O-hexadecyl-n-propylpropane-i, 3-diol-3-phosphoric acid 4 '- (1-quinuclidinium-butyl) ester (III with R ¹ = C ₁₆ H ₃₃ , R ¹ = nC ₃ H "R ₃ "-CH ₂ -CH ₂ -, Z = -CH = and η = 4) 557 mg = 1 mmol of dried I (R ₁ = 0-C ₁₄ H ₃₁₁ R ₂ = nC ₃ H ₇ , X = Br and η = 4) and 168mg = 1, SmOlII (R ¹ = -CH ₂ -CH ₂ -and

Z = -CH =) are dissolved in 8 ml of dried chloroform and in a sealed Gofäß on a water bath at 50 ° C.

tempered. After 3 hours, the TLC of the course of the reaction indicates complete reaction. The solution is cooled to room temperature, treated with 9 ml of methanol and 6 ml of water, shaken and then the organic phase evaporated to dryness in vacuo. The oily residue is dissolved in a mixture of 9 ml of methanol and 7.5 ml of chloroform, treated with 7.5 ml of 5% formic acid and shaken vigorously. The organic phase is separated, treated with 7.5 ml of sodium acetate solution (0.1 mmol / l) and 9 ml of methanol, shaken and then the organic phase evaporated to dryness in vacuo. The crude product is purified by column chromatography on 40 g of silica gel (MERCK) with chloroform / methanol / water (100: 30: 1) as Elutlonsmittel and dried in a desiccator over phosphorus pentoxide. 85% of the pure end product is obtained as a white, hygroscopic substance.

Elemental analysis: C ₃₃ H ₆₆ NO ₆ P χ H ₂ O (605.9)

Calcd .: N 2.31 P 5.11 Fe: N 2.25 P 4.92

Example 2

1-0- (Pentadecylcarbamoyl) -2-n-propylpropane-1,3-diol-3-phosphoric acid 4 '- (1-quinuclidinium) -butyl ester (III with R ¹ = CO-NH-C ₁₆ H ₃₁ , R ² = nC ₃ H ₇ , R ₃ = -CH ₂ -CH ₂ -, Z = -CH = and η = 4) 585 mg = 1 mmol of dried I (R ¹ = CO-NH-C ₁₆ H ₃₁₁ R ² = nC ₃ H ₇ , X = Br and η = 4) and 168 mg = 1.5 mmol Il (R ³ = -CH ₂ -CH ₂ -and Z = -CH =) are reacted analogously to Example 1. Obtained by column chromatography on 40g

Silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 1) as eluant 80% of the pure end product as

pale yellow, hygroscopic substance.

Elemental analysis: C ₃₃ H ₆₆ N ₂ O ₆ P x H ₂ O (634.9)

Calcd .: N, 4.41, P4.88, Ge: N, 4.46, P, 4.92

Example 3

iO-hexadecyl-n-propyl-propane-IS-diol-S-phosphoric acid-α'-d-quinuclidinium-pentyl ester (III where R ¹ = C ₁₁ H ₃₃ , R ² = nC ₃ H ₇ , R ³ = CH ₂ - CH ₇ -, Z = -CH = and η = 5) 571 mg = 1 mmol of dried I (R ¹ = C ₁₆ H ₃₃ , R ² = nC ₃ H ₇ , X = Br and η = 5) and 168 mg = 1 , 5 mmol Il (R ³ = -CH ₂ -CH ₂ -, Z = -CH =) are reacted analogously to Example 1. After purification by column chromatography on 40 g of silica gel 60 (MERCK) with chloroform /

Methanol / water (100: 30: 1) as eluent 85% of the pure end product as a white, hygroscopic substance

receive.

Elemental analysis: C ₃₄ H ₆₈ NO ₆ P x H ₂ O (619.9)

Calcd .: N 2.26 P4.50 Fe: N 2.18 P 4.61

Bet game 4

iO-hexadecyl-ethyl-propane-IS-diol-S-phosphoric acid-α'-d-quinuclidinium- ¹ -pentyl ester (III with R ¹ = C ₁₆ H ₃₃ , R ² = C ₂ H ₆ , R ³ = -CH ₂ -CH ₂ -, Z = -CH = and η = 5) 557 mg = 1 mmol dried I (R ¹ = C ₁₆ H ₃₃₁ R '= C ₂ H ₆₁ X = Br and n = 5) and 168 mg = 1.5 mmol Il (R ³ = -CH ₂ -CH ₂ -, Z = -CH =) are reacted analogously to Example 1. Ma obtained after column chromatography on 40g silica gel 60 (MERCK) with

Chloroform / methanol / water (100: 30: 1) as eluent 80% of the pure end product as a white, hygroscopic substance. Elemental analysis: C ₃₃ H ₆₆ NO ₆ P x H ₂ O (605.9)

8: N 2.31 P 5.11 Gf .: N 2.25 P 5.02

Example 5

1-0-hexadecyl-2-0-ethyl-glycero-3-phosphoric acid 6 '- (1-quinuclidinium) hexyl ester (III with R ¹ = C ₁₆ H ₃₃ , R ² = 0-C ₂ H ₆ , R ³ = -CH ₂ -CH ₂ -, Z = -CH = and n = 6) 587 mg = 1 mmol dried I (R ¹ = C ₁₆ H ₃₃₁ R ² = O-C ₂ H ₆₁ X = Br and η = 6) and 168 mg = 1.5 mmol Il (R ³ = -CH ₂ -CH ₂ -, Z = -CH =) are reacted analogously to Example '. After purification by column chromatography on 40 g of silica gel 60 (MERCK) with

Chloroform / methanol / water (100: 30: 1) as eluents become 90% of the pure end product as white, strong

obtained hygroscopic substance.

Elemental analysis: C _M H _M NO ₆ P x H ₂ O (631.9)

Calcd .: N, 2.21, P4.90, Fe: N, 2.30, P, 4.82

Examples

1-0-Hexadecyl-2-n-propylpropane-1,3-diol-3-phosphoric acid 8 '- (1-quinuclidinium) octyl ester (III with R ¹ = C ₁₆ H ₃₃ , R ² = nC ₃ H ₇ , R ³ = -CH ₂ -CH ₂ -, Z = -CH = and η = 8) 613 mg = 1 mmol of dried I (R ¹ = C ₁₆ H ₃₃ , R ² = nC ₃ H ₇ , X = Br and η = 8) and 168 mg = 1.5 mmol Il (R ³ = -CH ₂ -CH ₂ -, Z = -CH =) are reacted analogously to Example 1. After purification by column chromatography, 40 g of silica gel are obtained

(MERCK) with chloroform / methanol / water (100: 30: 1) as eluant 80% of the pure end product as a white, hygroscopic substance. Elemental analysis: C ₃₇ H ₇₄ NO ₆ P = H ₂ O (662.0) Boron: N 2.12 P 4.68 Fe: N 2.04 P 4.56

Example 7

1-O-hexadecyl-2-O-ethyl glycero-3-phosphoric acid 6'-4-aza-1-azoniabicyclo (2,2,2) octane) hexyl ester (III where R ¹ = C ₁₆ H ₃₃ , R ² = O-C ₂ H ₆ , R ₃ ^ CH ₂ -CH ₂ -, Z = N = and η = 6) 5P7mg = 1 mmol of dried I (R, = C ₁₆ H ₃₃ , R ₂ = 0- C ₂ H ₆ , X = Br and η = 6) and 168 mg = 1.5 mmol Il (R ³ = -CH ₂ -CH ₂ -Z = N =) are reacted analogously to Example 1. After purification by column chromatography on 40 g of Kieselgol 60 (MERCK) with chloroform /

Methanol / water (100: 70: 5) as eluent 90% of the pure end product as a colorless, hygroscopic substance

receive.

Elemental analysis: C ₃₃ He ₃ N ₂ O ₈ P x H ₂ O (632.9)

Re: N 4,43 P4,89Gef .: N 4,39 P4,77

Examples

iO-hexadecyl-O-ethylglycero-S-phosphoric acid e'-IN-methyl-morpholinium- ¹ -hexyl ester (III with R ¹ = C ₁₆ H ₃₃ , R ² = O-C ₂ H ₆ , R ₃ = CH ₃ , Z = -O- and n = 6) 587 mg = 1 mmol of dried I (R '= C ₁₈ H ₃₃₁ R ² = C ₂ H ₆ , X = Br and η = e) and 202 mg = 2 mmolll (R ³ = -CH ₃ , Z = -O-) are dissolved in 8 ml of dried chloroform and tempered in the closed vessel on a water bath for 30 h at 60 ° C. After this time, the thin-layer chromatographic control of the course of the reaction indicates extensive reaction. The solution is cooled to room temperature, treated with 9 ml of methanol and 6 ml of water, shaken, the organic phase separated and evaporated in vacuo to dryness. The residue is dissolved in a mixture of 9 ml of methanol and 7.5 ml of chloroform, treated with 7.5 ml of 5% formic acid and shaken vigorously. The organic phase is separated off, combined with 7.5 ml of sodium acetate solution (0.1 mmol / l) and 9 ml of methanol, shaken and then the organic phase is evaporated to dryness in vacuo. The crude product is purified by column chromatography on 40 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 50: 5) as eluent and dried in a desiccator over phosphorus pentoxide. This gives 50% of the pure end product as a white, hygroscopic Pulvor. Elemental analysis: C ₃₂ H ₆₈ NO ₇ P x H ₂ O (625.9) Calcd .: N 2.24 P 4.95 Ge: N 2.18 P 4.84

Example 9

iO-hexadecyl-O-ethylglycero-S-phosphoric acid-o'-IN-methylpiperidinium-pentyl ester (III with R ¹ = C ₁₆ H ₃₃ , R ² = O-C ₂ H ₆ , R ³ = CH ₃ , Z = -CH ₂ - and η = 5) 575 mg = 1 mmol dried I (R ¹ = C ₁₆ H ₃₃ , R ² = O-C ₂ H ₆ , X = Br and η = 5) and 198 mg = 2 mmol Il (F! ³ = -CH ₃ , Z = -CH ₂ -) are reacted analogously to Example 8. After purification by column chromatography on 40 g of silica gel 60 (MERCK) with chloroform /

Methanol / water (100: 30: 2) as eluent 60% of the pure end product as a white, hygroscopic substance

receive.

Elemental Analysis: C ₃₂ H ₆₈ NO ₈ Px H ₂ O (610.9)

Calcd .: N, 2.29, P, 5.07G, N, 2.33, P, 5.15

Example 10

iO-hexadacyl-O-ethyl-glycero-S-phosphoric acid-o'-IN-methyl-piperidinium- ¹ -hexyl ester (III with R ¹ = C ₁₀ H ₃₃ , R ² = O-C ₂ H ₅ , R ³ = CH ₃ , Z = -CH ₂ - and η = 6) 587 mg = 1 mmol dried I (R ¹ = C ₁₆ H ₃₃ , R ² = O-C ₂ H ₅ , X = Br and η = 6) and 198 mg = 2 mmol Il (R ³ = -CH ₃ , Z = -CH ₂ -) are reacted analogously to Example 8. After purification by column chromatography, 40 g of silica gel 60 (MERCK) are obtained

Chloroform / methanol / water (100: 30: 2) as eluent 60% of the pure end product as a white, hygroscopic substance. Elemental analysis: C ₃₁ H ₆₄ NO ₆ P x H ₂ O (595.8)

Calc .: N 2.35 P 5.20 Fe: N 2.29 P 5.12

Example 11

iO-hexadecyl-n-propyl-propane-IS-diol-S-phosphoric acid-eMN-methylpiperidinium-octyl ester (III with R ¹ = C ₁₆ H ₃ -, R ² = nC ₃ H ₇ , R ³ = -CH ₃ , Z = -CH ₂ - and η = 8) 613 mg = 1 mmol of dried I (R '= C ₁₆ H ₃ -R ² = nC ₃ H ₇ , X = Br and η = 8) and 198 mg = 2 mmol of Il (R ³ = -CH ₃ , Z = -CH ₂ -) are reacted analogously to Example 8. After purification by column chromatography on 40 g of silica gel 60 (MERCK) with chloroform /

Methanol / water (100: 30: 2) as the eluent, 65% of the pure Endprodul; t6S as a white, hygroscopic substance

receive.

Elemental analysis: C ₃₆ H ₇₄ NO ₅ P χ H ₂ O (650.0)

Calcd .: N 2.15 P4.77 Ge: N 2.20 P4.86

Characterization of the PAF antagonistic effect

The antagonistic action of the compounds according to the invention (general formula III) is described below using the example of the inhibition of PAF-induced platelet aggregation. The latter was carried out in human platelet citrated plasma by the method described by BORN (J. Physolol. (London) 162, 67-68 (19621) using an ELVI840 two-channel aggregometer (Elvi Logos, Milan, Italy) in the presence of the compounds of the invention If one observes a concentration-dependent inhibition of PAF-mediated platelet aggregation, this can be reversed by increasing the PAF concentration.SHILD analysis of the observed parallel shift in the concentration-response curve shows that the compounds of the invention act as competitive antagonists of PAF For the effectiveness, the corresponding equilibrium dissociation constants (Ke values, see Table 1) were determined.

In contrast to PAF, the thrombocyte aggregation induced by other triggers, such as ADP, collagen or arachidonic acid, is only insignificantly influenced by the compounds according to the invention.

Table 1: Ke values experimentally determined on human platelets

Verb. R ¹ R ² R ³ Z n K ₈ -WeH (minor) Ill C, ₉ H ₃₃ nC ₃ H, -CH ₂ -CH ₂ - CH = 4 5.4 x ΙΟ "' III CONH-C ₁₆ H ₃ , nC ₃ H, -CH ₂ -CH ₂ - CH = 4 4.1 x ΙΟ "' III Ci ₈ H ₃₃ nC ₃ H, -CH ₂ -CH ₂ - CH = 5 3.4 x ΙΟ " ⁶ III Ct ₈ H ₃₃ C ₂ H ₆ -CH ₂ -CH ₂ - CH = 5 4,3x10- ' III Ci ₈ H ₃₃ 0-C ₂ H ₆ -CH ₂ -CH ₂ - CH = 6 1,1X10- " III CIh ₃₃ nC ₃ H, -CH ₂ -CH ₂ - CH = 8th > 10 " ⁶ III C ₈ H ₃₃ 0-C ₂ H ₆ -CH ₂ -CH ₂ - -N = 6 > 10 " ⁶ III C) ₈ H ₃₃ 0-C ₂ H ₆ -CH ₃ -0- 6 > 1 (T ⁶ III CieH ₃₃ 0-C ₂ H ₆ -CH ₃ -CH ₂ - 5 3,9X10 "" III CeH ₃₃ 0-C ₂ H ₆ -CH ₃ -CH ₂ - 6 2,3 x 10 " ⁶ III Ci ₈ H ₃₃ . nC ₃ H ₇ -CH ₃ -CH ₂ - 8th > 10 " ⁶

Claims (1)

1. A process for the preparation of novel, effective PAF antagonists with phospholipid structure, characterized in that starting compounds of the general formula I. CII-R > - X OO ^w wherein R ^{1 represents} a long-chain, unsubstituted alkyl or alkylcarbamoyl radical having 16 to 20 carbon atoms, for R ^{2 is} a methoxy, ethoxy, propoxy or benzoxy group or a methyl, ethyl or n-propyl to n-pentyl radical or a hydroxy group stands for η are the numbers 4 to 10 and X is chlorine or bromine, with acyclic tertiary bases of the general formula II,