DD279677A1 - METHOD FOR PRODUCING NEW, HIGHLY ACUTE ANTAGONISTS OF THE PLAETTE ACTIVATING FACTOR WITH PHOSPHOLIPID STRUCTURE - Google Patents

Abstract

The invention relates to a process for the preparation of previously not described phospholipids and their characterization as highly effective antagonists of the platelet-activating factor. The aim of the invention is to produce by a suitable method new PAF antagonists with high efficiency, good water solubility and plasma stability. This object is achieved by reacting haloesters of phospholipids with various pyridines by a simple process. 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.

Description

Characterization of the known technical solutions

Since PAF's structure elucidation in 1979 and its characterization as a physiological mediator of allergic and inflammatory reactions, PAF antagoniston has become increasingly interesting as potential agents and drugs. The currently known antagonists can be divided into three groups:

1. Natural anti-PAF antagonists such as Ginkgo biloba BN 52020-52023 (BRAQUET, GB Pat. 8418424 [19841; Ginkgolides Chemistry, Biology, Pharmacology, and Clinical Perspectives, P. BRAQUET [Ed.], JR Prous Science Publishers, Barcelona 1987), benzoiuranoide neolignans such as Kadsurenon from Piper futokadsurae (SHEN et al., Proc Nat, Acad., Sci., USA, 82, 672 [1985]), 2,3,4,5-tetrasubstituted furanoid lignans such as Veraguenesin from Magnolia acuminata (SHEN et al., 2nd World Conference on Inflammation, Monte Carlo 1986), Galgravin and Galbelgin from Himantandra belgravena (BRAQUET et al., Trends Biochem., Be.7, 397 [1986]), 3,4-disubstituted furanoid lignans, such as Buseran from Busera microphylla (BRAQUET et al., Ibid), butanolide lignans, such as Prestegan from Steganotaenia aralisces (BRAQUET et al., New Platens in Platelet Activating Factor Research, ML LEE and CM WINSLOW [Ed.], John Wiley & Sons, Chichester 1987), and US Pat microorganisms isolated gliotoxin derivatives FR-4 91/5 and FR-900452 (OKAMOTO et al., Chem. Pharm. Bull. 34, 340 [1986], OKAMOTO et al. J.

Antibiotics 39, 188 [1986]).

2. Synthetic heterocycles such as a series of triazolobenzodiazepines (KORNECKI et al., Science 226, 1454 [1984], CASALS-STENZEL et al. J. Pharmacol. Expt. Ther.241,974 [1987]), 48740 RP, a (3-pyridyl) -1 H.SH-pyrrolo-d1-ci-thiazolium derivative (FARBE et al., Eur. Pat., 115979 [1983]), indene derivatives (SHEN et Eur. Pat. 142801 (19831) and 2,5-diaryltetrahydrofurans and analogous compounds such as L-652731 and L-653150 (BIFTU et al., Eur. Pat., 144804 [1983])

3. PAF structural analogues such as CV-3988, which contains an octadecylcarbamoyl radical in the C-1 position, a methoxy radical in the C-2 position, and a thiazoliumethyl-head group in the C-3 position of the glycerol (TERASHITA et al., Life Sci., 32, 1975 [1983 ]), ONO-6240, an iO-hexadecyl ^ -O-ethyl-glycero-SO-heptamethylenethiazolium salt (MIYAMOTO et al., Eur. Pat. 146258 [1985]), RO-193704, RO-188736 and RO-187953, inv A series of glycerolipids having an octadecylcarbamoyl radical in the C-1 position, a methoxy or a methoxycarbonyip, at the C-2 position and a phosphate-free, ether-bonded thiazolium butyl head group (WINSLOW et al., Prostaglandins 30,697 [1985]), RU-45703 also represents a 1-0-octadecyl-2-0-ethylglycerolipid and is esterified in the C-3 position with a butylcholine residue (WICHROWSKI et al., 6th International Conference on Prostaglandins and Related Compounds, Florence 1986).

Furthermore, several glycerol-free backbone antagonists derived from the PAF structure have been described, including SRI-63073, which is characterized by a piperidine backbone and a thiamine phosphate head group (LEE et al., Prostaglandins 30,690 [1985]) and derivatives having a dioxanone backbone and a Alkylpyridinium head group (BURRI et al., Prostaglandins 30,691 [1985]).

Most of the previously described natural products and heterocycles with PAF-antagonistic activity are sparingly soluble in water and for this reason are of limited use for pharmacological purposes. On the other hand, their isolation and synthesis are usually associated with extremely high experimental effort and provide the target compounds usually only in low yields, so that they meet the requirements of a technically applicable method insufficient. Also, the described antagonistic PAF structural analogs are generally poorly water-soluble (CV-3988, SRI-63119, and the RO series), have been rapidly degraded in plasma (RO-19400), and are currently of low toxicity. only a low potency (SRI-63119 and SRI-63073). In addition, many of these compounds have a significant procedural disadvantage in the low chemical reactivity of the thiazole to be introduced as head group (CV-3988, ONO-6240, SRI-63119 and the RO series), whereby only low yields are achieved despite the use of drastic reaction conditions.

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, highly effective PAF antagonists. The starting compounds are haloesters of phospholipids which can be prepared in a manner known per se.

According to the invention the object is achieved in that starting compounds of general formula I,

₀ -R ¹

CH-R _(|)

CH ₀ OPO- (CH ₀ ) -X 2 // \ 2 τι

OH

wherein R ^{1 represents} an unsubstituted, long-chain alkoxy radical having 12 to 20 carbon atoms, R ^{2 represents} a methoxy, ethoxy, propoxy or benzoxy group or a methyl, ethyl or n-propyl to n-penty! rest or a hydroxy group , R ¹ and R ^{2 may} also be interchanged, for η are the numbers 2 to 10 and X is chlorine, bromine and iodine, with ortho, meta, or para-substituted pyridines of the general formula II,

(II)

wherein R ^{3 represents} an amino group which may be substituted by identical or different n-alkyl radicals having 1 to 4 carbon atoms, which together may also form a 3- to 7-membered ring, to compounds of general formula III,

CH ₀ OPO

² / A

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

The reactions are carried out in anhydrous halogenated hydrocarbons, such as methylene chloride, chloroform, trichlorethylene or in anhydrous solvent mixtures of chloroform / acetonitrile (1: 1) with exclusion of moisture. To increase the yield, the amount of the substituted pyridine is preferably carried out in a sealed vessel in the temperature range of 4O ⁰ C to 60 ° C. Compounds of the general forms! Ill, where R ¹ , R ³ and η have the abovementioned meaning and R ² is OH and R ¹ and R ^{2 can} also be interchanged, are prepared with suitable derivatives of short-chain carboxylic acids, such as acetic acid or propynoic acid, to give compounds of the general formula IV ₁

(IV)

O-P-O-CCH

wherein R ¹ , R ³ and η are as defined above and R ^{4 is} an acetyloxy or Propionyloxyr reacted. Suitable derivatives are acid anhydrides or acid chlorides of said carboxylic acids. The reactions are carried out in anhydrous organic solvents such as pyridine, 1 riethyl amine or quinoline and in Losungsmi'tnlgemischen of chloroform / pyridine or chloroform / triathylamine (1: 1). Preference is given to using pyridine as solvent and an 8 molar excess of acid anhydride as acylating reagent. The reactions are carried out in the temperature range from 2O ⁰ C to 4O 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. The compounds according to the invention thus represent a valuable enrichment of the active compounds known in the field. The invention will be explained below using Alisführungsbeispielen.

example 1

i-O-hexadecyl-O-ethylglycero-S-phosphoric acid WDimethylamino-pyridinium ethyl ester [III with R '= O-C ₁₆ H ₃₃ , R ² = O-C ₂ H ₅ , R ₃ = pN (CH ₃ ) ₂ and η = 2

531 mg = 1 mmol dried I (Ri = O-C ₁₆ H ₃₃ , R ₂ = O-C ₂ H ₅ , X = Br and η - 2) uni '45mg = 2 mmol Il (R ³ = pN (CH ₃ ) ₂ | are dissolved in 7.5 ml chloroform and dried in a closed vessel i- _u i of the water bath at 50 ° C tempered After 12 hours the thin layer chromatographic control indicates complete reaction the solution is cooled to room temperature, diluted with 9 ml of methanol and.. 5 ml of water, shaken and then the organic phase evaporated to dryness in vacuo.The remaining 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 Phase is separated off, mixed with 7.5 ml of sodium acetate solution (0.1 mmol / l) and 9 ml of methanol, and then evaporated to dryness in vacuo The crude product is purified by column chromatography on 20 g of silica gel 60 (MEflCK) with chloroform / methanol / water (100: 1). 30: 2) as eluent el cleaned and dried in a desiccator over phosphorus pentoxide. 70% of the pure end product is obtained as a white, strongly hygroscopic substance

Elemental analysis: C ₃₀ H ₆₇ N ₂ O ₆ P · H ₂ O (590.5)

Calc .: N 4.75 P 5.24

Gef.:N4,83P5,10

Example 2

iO-hexadecyl ^ -n-propylglycero-S-phosphoric acid ^ -idymethylaminol-pyridinium ethyl ester (III with R '= O-C ₁₆ H ₃₃ , R ² = OnC ₃ H ₇ , R ₃ = pN (CH ₃ ) ₂ and η = 2]

545 mg = 1 mmol of dried I [(R ¹ = O-C ₁₆ H ₃₃ , R ² = OnC ₃ H ₇ , C = Br and η = 2) and 245 mg = 2 mmol of Il [R ³ = pN (CH ₃ ) ₂ ] are reacted analogously to Example 1. After purification by column chromatography on 20 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluent, 65% of the pure end product is obtained as white, strongly hygroscopic Substaru.

Elemental analysis: C 1 H ₅₉ N ₂ O ₆ P · H ₂ O (604.5)

Ber .: ι \ Μ.β4Ρ5,12

Gef.:N4,56P5,01

Example 3

iO-Dodecyl-O-ethylglycero-S-phosphoric acid ^ -dimethylamino-1-pyridinium ethyl ester [III with R '= O-C ₁₂ H ₂₅ , R ² = O-C ₂ H ₅ , R ³ = DN (CHj) ₂ and n = 2)

475 mg = 1 mmol of dried I [R '= O-C ₁₂ H ₂₅ , R ² = O-C ₂ H ₅₁ X = Br and η = 2) and 245 mg = 2 mmol [R ³ = pN (CH ₃ ) ₂ ] implemented analogously to Example 1. After purification by column chromatography on 20 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluent, 70% of the pure end product is obtained in the form of a white, strongly hygroscopic substance.

Elemental Analysis: C ₂₆ H ₄₉ N ₂ O ₆ PH ₂ O (534.4)

Calc .: N5,24 P5,80

Found: N 5.32 P 5.68

Example 4

iO-hexadecyl-O-ethyl-glycero-S-phosphoric acid, -IV-aminol-pyridinium ethyl ester (III with R '= O-C ₁₆ H ₃₃ , R ² = O-C ₂ H ₅ , R ³ = p-NH ₂ and η = 21

531mg = 1mmol. Rocknets I [R '= O-C ₁₆ H ₃₃ , R ² = O-C ₂ H ₅ , X = Br and η = 2 I and 188 mg = 2mmoiriR ³ = D-NH ₂ ) will be under moderate L. warming dissolved in 7.5 ml of dried chloroform and then heated in a closed vessel on the water at 4O ⁰ C. After 8 hours, the TLC shows complete conversion. The cloudy solution is cooled to room temperature, filtered and then treated with 9 ml of methanol and 5 ml of water. The mixture is shaken vigorously, the organic phase separated and evaporated to dryness in vacuo. The remaining 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, shaken vigorously and the organic phase separated. This is mixed with 7.5 ml of sodium acetate solution (0.1 mmol / l) and 9 ml of methanol, shaken, separated and evaporated in vacuo. For purification, the residue is separated on 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 35: 3) as eluent and dried in a desiccator over phosphorus pentoxide. 75% of the pure end product is obtained as white analytically pure substance. Elemental analysis: C ₂₈ H ₆₃ N ₂ O ₆ P · H ₂ O (562.5)

Calc .: N4.98 P5.51

Gef .: N4,87 P5,58

Example 5

iO-hexadecyl ^ -n-propylglycero-S-phosphoric acid ^ -fAminol-pyridinium ethyl ester [III with R '= O-C ₁₆ H ₃₃ , R ² = OnC ₃ H ₇ , R ³ = p-NH ₂ and η = 2]

545mg = 1mmo! dried I [R '= O-C ₁₆ H ₃₃₁ R ² = OnC ₃ H ₇ , X - Brundn = 2 I and 188 mg = 2 mmolll [R ³ = p-NH ₂ ] are reacted analogously to Example 4. After purification by column chromatography on 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 35: o) as eluent, 70% of the pure end product is obtained as white crystalline substance.

Elemental analysis: C ₂₉ H ₆₅ N ₂ O ₆ PH ₂ O (576.5)

Calc .: N4.86 P5.37

Gef.:N4,73P5,31

Example 6

iO-hexadecyl-O-ethylglycero-S-phosphoric acid IDimethylaminol-pyridinium propyl ester [III with R ¹ = O-C ₁₆ H ₃₃ , R ² = O-C ₂ H ₆ , R ³ = pN (CH ₃ ) ₂ and η = 3] 545 mg = 1 mmol of dried I | R '= O-C ₁₆ H ₃₃ , R ² = O-C ₂ H ₅ , C = Br and η = 3 | and 245mg Il = (R ³ = 2 mmol pN (CH _{3) 2]} dissolved in 7.5 ml of dried temperature-trichlorethylene and in a closed vessel in the water bath at 6O ⁰ C. After 10 hours, the thin-layer chromatographic control indicates complete reaction. The The solution is cooled to room temperature, admixed with a mixture of 9 ml of methanol and 5 ml of water, shaken, and then the organic phase is evaporated to dryness in vacuo The white residue is dissolved in a mixture of 9 ml of methanol and 7.5 ml of chloroform, with 7, 5 ml of 5% formic acid are added, shaken vigorously and the organic phase is separated off, treated with 7.5 ml of sodium acetate solution (0.1 mmol / l) and 9 ml of methanol, shaken, separated and evaporated in vacuo Silica gel 60 (MERCK) was purified by means of chloroform / methanol / water (100: 30: 2) as eluent and dried in a desiccator over phosphorus pentoxici n receives 75% of the pure end product as a white, hygroscopic substance. Elemental analysis: C ₃₁ H ₅₉ N ₂ O ₆ P · H ₂ O (604.5) Ber.:N4,64P5,12 Gef.:N4,72P5,17

Example 7

iO-hexadecyl-O-ethyl-glycero-S-phosphoric acid-dimethylaminopyridinium-butyl ester [III with R ¹ = O-C ₁₆ H ₃₃ , R ² = O-C ₂ H ₅ , R ³ = pN (CH ₃ ) ₂ and n = 4 ] 559 mg = 1 mmol of dried I [R ¹ = O-C ₁₆ H ₃₃₁ R ² = OC ₂ H ₅ , X = Br and η = 4] and 245 mg = 2 mmol [R ³ = DN (CH ₃ I ₂ ] are analogous Example 6 After purification by column chromatography on 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluent, 70% of the pure end product is obtained as a white hygroscopic substance.

Elemental analysis: C ₃₂ H ₆₁ N ₂ O ₆ P · H ₂ O (618.5) Calc. N 4, 53 P 5.01 Found: N 4.44 P 4.94

Example 8

iO-hexadecyl ^ -n-propylglycero-S-phosphoric acid ^ -imidylamino- ¹ -pyridinium-butyl ester [III with R ¹ = O-C ₁₆ H ₃₃ , R ² = OnC ₃ H ₇ , R ³ = pN (CH ₃ ) ₂ and η = 4) 573 mg = 1 mmol of dried I [R '= O-C ₁₆ H ₃₃₁ R ² = OnC ₃ H ₇ , X = Br and η = 4] and 245 mg = 2 nm molll [R ³ = pN (CH ₃ ) ₂ ] implemented analogously to Example 6. After purification by column chromatography on 25 g of silica gel CO (MERCK) with chloroform / methanol / water (100: 30: 2) as eluent, 70% of the pure end product is obtained as a white, slightly hygroscopic substance.

Elemental analysis: C ₃₃ H ₆₃ N ₂ O ₆ P · H ₂ O (632.5) Ber.:N4,43P4,90 Gef.:N4,63P4,81

Example 9

iO-hexadecyl-O-n-propylglycero-S-phosphoric acid ^ dimethyldimethylpyridinium pentyl ester [III with R '= O-C ₁₆ H ₃₃ , R ² = OnC ₃ H ₇ , R ³ = pN (CH ₃ ) ₂ and η = 5 ) 587 mg = 1 mmol of dried I [R ¹ = O-C ₁₆ H ₃₃₁ R ² = OnC ₃ H ₇ , X = Br and η = 5) and 245 mg = 2 mmol [R ³ = pN (CH ₃ ) ₂ ] are analogous Example 6 implemented. After purification by column chromatography on 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluant, 75% of the pure end product is obtained as a white, slightly hygroscopic substance.

Elemental analysis: C ₃₄ H ₆₅ N ₂ O ₆ P · H ₂ O (646.5) Ber.:N4,33P4,79 Gef.:N4,43P4,71

Example 10

iO-hexadecyl-O-ethyl-glycero-S-phosphoric acid ^ -dimethylamino- ¹ -pyridinium pentyl ester [III with R ¹ = O-C ₁₆ H ₃₃ , R ² = O-C ₂ H ₅ , R ³ = pN (CH ₃ ) ₂ and η = 5] 573 mg = 1 mmol of dried I [R '= O-C ₁₆ H ₃₃ , R ² = O-C ₂ H ₅ , X-Br and η = 5! and 245 mg = 2 mmol of Il [R ³ = pN (CH ₃ ) ₂ ] are reacted analogously to Example 6. After purification by column chromatography on 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluant, 75% of the pure end product is obtained as a white, slightly hygroscopic substance.

Elemental analysis: C ₃₃ H ₆₃ N ₂ O ₆ P H ₂ O (33I ', 5) Ber.:N4,43P4,90 Gef.:N4,48P4,95

Example 11

i-O-hexadecyl-O-ethylglycero-S-phosphoric acid ^ -idymethylamino-1-pyridinium hexyl ester Uli where R '= O-C ₁₆ H ₃₃ , R' = O-C ₂ H ₆ , R ³ = pN (CH ₃ ) ₂ and η = 6] 587 mg = 1 mmol dried I [R '= O-C ₁₆ H ₃₃₁ R ² = O-C ₂ H ₅₁ X = Br and η = 61 and 245 mg = 2 mmolll [R ³ = pN (CH ₃ ) ₂ ] are reacted analogously to Example 6. After purification by column chromatography on 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluant, 70% of the pure end product is obtained as a white, slightly hygroscopic powder.

Elemental analysis: C ₃₄ H ₆₅ N ₂ O ₆ P · H ₂ O (646.5) Ber.:N, 4.33 P 4.79 Found: N, 4.29, P 4.64

Example 12

i-O-hexadecyl-O-propyl-propanediol-S-phosphoric acid, -ID-dimethylaminol-pyridinium ethyl ester [III with R ¹ = O-C ₁₆ H ₃₃ , R ² = nC ₃ H ₇ , R ³ = pN (CH ₃ ) ₂ and η = 529mg = 1mmo! dried I [R '= O-C ₁₆ H ₁₁ , R ² = nC ₃ H ₇ , X = Br and η = 2] and 24 Bmg = 2 mmolll [R ³ = pN (CH ₃ ) ₂ ] are reacted analogously to Example 1. Purification by column chromatography on 30 g of silica gel 60 (Merck) using chloroform / methanol / water (100: 30: 2) as eluent 70% of the pure ^r ndproduktes are obtained as white, highly hygroscopic substance.

Elemental analysis: C ₃₁ H ₅₉ N ₂ O ₆ P · H ₂ O (588.5) Ben: N, 4.76, P, 5.26 Found: N, 4.86, P, 5.13

Example 13

iO-hexadecyl-n-propyl-propanediol-S-phosphoric acid, -ID-dimethylaminol-pyridinium-propylster [III with R '= O-C ₁₆ H ₃₃ , R ² = nC ₃ H ₇ , R ³ = pN (CH ₃ ) ₂ and η = 3] 543mg = 1mmol dried I [R ¹ = 0-C ₁₆ Hj ₃ , R ² = nC ₃ H ₇ , X = Br and η = 3] and 245mg = 2mmolll [R ³ = DN (CH ₃ I ₂ ] The mixture is purified by column chromatography on 30 g silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluent 70% of the pure end product as white, strongly hygroscopic substance.

Elemental analysis: C ₃₂ H ₆₁ N ₂ O ₅ P · H ₂ O (602.3) Calc. N 4 .65 P 5.14 Found: N 4.58 P 5.36

Example 14

i-O-hexadecyl-n-propyl-propanediol-S-phosphoric acid-dimethylaminopyridinium-butyl ester [III with R ¹ = O-C ₁₆ H ₃₃ , R ² = nC ₃ H ₇ , R ³ = pN (CH ₃ ) ₂ and η = 4 ] 557 mg = 1 mmol dried I [R ¹ = O-Ci ₆ H ₃₃ , R ² = nC ₃ H ₇ , X = Br and η = 41 and 245 mg = 2m, oil Il [R ³ = pN (CH ₃ ) ₂ ) are reacted analogously to Example 6. After purification by column chromatography on 30 g of Kieselgci 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluent, 75% of the pure end product is obtained as a white, hygroscopic substance.

Elemental analysis: C ₃₃ H ₆₃ N ₂ O ₅ P · H ₂ O (616.3) Calc .: N 4.54 P 5.03 Gef.:N4,47P5,14

Example 15

iO-hexadecyl ^ -n-propylpropandiol-S-phosphoric ^ -fDimethylaminol-pyridiniumoctylester [Ml with R '= 0-Ci ₆ H _33, R ² = nC ₃ H _7, R ³ = pN (CH _{3) 2} and η = 8) 613 mg = 1 mmol of dried I [R '= O-C ₁₆ H ₃₃ , R ² = nC ₃ H ₇ , X = Br and η = 81 and 245 mg = 2 mmol of Il [R ³ = pN (CH ₃ ) ₂ ) are reacted analogously to Example 6. After purification by column chromatography on 30 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluent, 75% of the pure end product is obtained as a white, hygroscopic substance.

Elemental analysis: C ₃₇ H ₇₁ N ₂ O ₅ P · H ₂ O (672.2) Found .: Ber.:N4,17P4,61 N4,12P4,56

Example 16

iO-hexadecyl-n-pentylpropanediol-S-phosphoric acid ^ -d-dimethylaminol-pyridinium ethyl ester [III with R '= O-C ₁₆ H ₃₃ , R ² = n-CsH ,,, R ³ = pN (CH ₃ ) ₂ and η = 2] 557mg = 1 mmol dried I [R '0-Ci ₆ H ₃₃₁ R ² = nC ₅ Hn, X = Br and η = 2 | and 245 mg = 2 mmol of Il [R ³ = DN (CH ₃ I ₂ I are reacted analogously to Example 1. After purification by column chromatography on 30 g of silica gel 60 (Merck) with chloroform / methanol / water (100: 30: 2) as eluent 70% of the pure end product as white, strongly hygroscopic substance.

Elemental analysis: C ₃₃ H ₆₃ N ₂ O ₅ P · H ₂ O (616.6) Calcd .: N4,54 P5,02 Gef.:N4,64P5,19

Example 17

iO-hexadecyl ^ -O-benzylglycero-S-phosphoric ^ -IDimethylaminol-pyridiniumethylester [III where R '= 0-Ci ₆ H _33, R ² = O-CH _{3) 2} 1 and n = 2) = 1 mmol 593mg dried I [R ¹ = O-C ₆ H ₃₃ , R ² = O-CH ₂ -C ₆ H ₅ , X = Br and η = 2] and 245 mg = 2 mmolll [R ³ = DN (CH ₃ I ₂ I become analog Example 1 Reacted by column chromatography on 30 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 2 ', as eluent 50% of the pure end product as a colorless, highly hygroscopic substance.

Elemental analysis: C ₃₅ H ₅₉ N ₂ O ₆ P · H ₂ O (652.5) Calc. N 4, 29 P 4.74 Found: N, 4.22 P 4.62

Example 18

i-O-hexadecyl ^ -O-ethylglycero-S-phosphoric ^ -fPiperidinoi-pyridiniumpentylester

[III with R '= O-C ₁₆ H ₃₃ , R' = O-C ₂ H ₅ , R ³ = P "H (CH ₂ ) ε _{and n =} 5]

573 mg = 1 mmol of dried I [R ¹ = O-C ₁₆ H ₃₃ , R ² = O-C ₂ H ₅ , X = Br and η = 51 and 325 mg = 2 mmolll [R ³ = pI (CH ₂ ) si]

are dissolved in 7.5 ml of chloroform and heated in the closed vessel on the water at 55 ⁰ C. After 10 hours the TLC shows complete reaction. The solution is cooled to room temperature, treated with a mixture of 9 ml of methanol and 5 ml of water and shaken. The organic phase is taken up 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 separated organic phase is treated with 7.5 ml of sodium acetate solution (0.1 mmol / l) and 9 ml of methanol, shaken and then evaporated in vacuo. The resulting crude product is purified by column chromatography ar 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100/25/1) as the eluent and dried in a desiccator over phosphorus pentoxide. 75% of the pure end product is obtained as a white hygroscopic substance. Elemental analysis: C ₃₆ H ₆₇ N ₂ O ₆ P · H ₂ O (672.5) Ber.:N4,16P4,61 Gef.:N4,10P4,73

Example 19 i-O-Hexadecyl ^ -O-ethylglycero-S-phosphoric acid ^ -fPyrrolidinol pyridinium pentyl ester

[III with R ¹ = 0-C ₁₆ H ₃₃ , R ² = 0-C ₂ H ₆ , R ³ = p-ff (CHs) ₄ and η = 5)

673 mg = 1 mmol dried I [R ¹ = O-C ₁₆ H ₃₃ , R ² = O-C ₂ H ₅ , X = Br and η - 5 | and 325mg = 2mm ο I Il

are implemented analogously to Example 18. After purification by column chromatography on 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 25: 1) as eluent, 75% of the pure end product is obtained as a white, hygroscopic substance. Elemental analysis: C ₃₅ H ₆₅ N ₂ O ₆ PH ₂ O (658.5) Ber.:N4,25P4,70 Gef.:N4,14P4,82

Example 20

iO-hexadecylglycero-S-phosphoric acid ^ -imidylamino-1-pyridinium ethyl ester [III with R '= O-C ₁₆ H ₃₃ , R ² = pN (CH ₃ ) ₂ and n = 2] 503 mg - 1 mmol of dried I [R ¹ = 0-C ₁₆ H ₃₃₁ R ² = OH, X = Br and η = 2] and 245 mg = 2 mmol of Il [fl ³ = pN (CH ₃ ) ₂ ] are reacted analogously to Example 1. After purification by column chromatography on 25 g of Kieselge ¹ 60 (MERCK) with chloroform / methanol / water (100: 35: 3) as eluent, 65% of the pure end product is obtained as a colorless, strongly hygroscopic substance.

Elemental analysis: C ₂₈ H ₅₃ N ₂ O ₆ PH ₂ O (562.5) Calc .: N 4.98 P 5.51 Found .: N 4.84 P 5.55

Example 21

2-O-hexadecylglycero-3-phosphoric acid ^/ 1- (dimethy ^: mino) -pyridinium ethyl ester [III with R ¹ = OH. R ² = O-C ₁₆ H ₃₃ , R ³ = pN (CH ₃ ) ₂ and = 2] 503 mg = 1 mmol of dried I [R '= OH, R ² = O-, ₁₆ H ₃₃ , X = Br and η = 2] and 245 mg = 2 mmolll [R ³ = pN (CH ₃ ) ₂ l are analogous to Example 1 implemented. After purification by column chromatography on 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 35: 3) as eluent, 70% of the pure end product is obtained as a colorless, starV hygroscopic substance.

Elemental analysis: C ₂₈ H ₅₃ N ₂ O ₆ P · H ₂ O (562.5) Calcd .: N4,98 P5,51 Gef.:N4,93P5,64

Example 22

i-O-benzyl-O-hexadecylglycero-S-phosphoric acid, -ID-dimethylaminol-pyridinium ethyl ester [III with R '= O-CH ₂ -C ₆ H ₆ , R ² = O-C ₁₆ H ₃₃ , R ³ = pN (CH ₃ ) ₂ and η = 2) 593 mg = 1 mmol of dried I [R ¹ = O-CH ₂ -C ₆ H ₅ , R ² = O-C ₁₆ H ₃₃ , X = Br and η = 2] and 245 mg = 2 mmol of II [R ³ = pN (CH ₃ ) ₂ ] are reacted analogously to Example 1. After purification by column chromatography on 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 30/2) as eluant, 55% of the pure end product is obtained as a colorless, strongly hygroscopic substance.

Elemental analysis: C ₃₅ H ₅₉ N ₂ O ₆ PH ₂ O (652.5) Ber.:N4,29P4,74 Gef.:N4,34P4,82

Example 23

i-O-ethyl-S ^ -O-hexadexylglycero-phosphoric ^ -fDimethylaminol-pyridiniumpropylester

(III with R '= O-C ₂ H ₅ , R ² = O-C ₁₆ H ₃₃ , R ³ = pN (CH ₃ ) ₂ and n = 3]

545 mg = 1 mmol of dried I (R ¹ = OC ₂ H _E , R ² - 0-C ₁₆ H ₃₃ , X = Br and n = 3) and 245 mg = 2 mmol of Il [R ³ = pN (CH ₃ ) ₂ ] are reacted analogously to Example 6. After column chromatography purification on 25 g of silica gel 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluent 75% of pure

End product as a white, hygroscopic substance.

Elemental analysis: C ₃₁ H ₅₉ N ₂ O ₆ P · H ₂ O (604.5)

Ber .: N4,64P5,12

Gef.:N4,58P5,22

Example 24

i-O-hexadecyl ^ -O-acetylglycero-S-phosphoric ^ -fDimethylamionl-pyridiniumpropylester

[IV with R ¹ = O-C ₁₆ H ₃₃ , R ³ = pN (CH ₃ ) ₂ , R ⁴ = O-CO-CH ₃ and n = 3]

87mg = 0.15mmol dried III [R '= O-C ₁₆ H ₃₃₁ R ³ = pN (CH ₃ ) ₂ , R ⁴ = OH and η = 3] are suspended in 190mg = 2.4mmol of dried pyridine. 122 mg = 1.2 mmol acetic anhydride are added and the temperature is maintained in the sealed flask at 40 ° C. for 8 hours. After this time, the thin-layer chromatography indicates almost complete conversion. Are added 2 ml of water is added and heated for another hour at 4O ⁰ C. The Reaktioni jemisch is left to stand overnight and then iyophibilisiert. The residue is purified by column chromatography on 15 g of Kieselgol 60 (MERCK) with chloroform / methanol / water (100: 30: 2) as eluent. 60% of the pure end product is obtained as white, strong

hygroscopic substance.

Elemental analysis: C ₃₁ H ₅₇ N ₂ O ₇ PH ₂ O (618.8)

Calc .: N4,52 P5,00

Found .: N 4,48 P 4,92

Example 25

i-O-acetyl ^ -O-hexadecylglycero-S-phosphoric ^ -IDimethylaminol-pyridiniumethylester

[IV with R ¹ = O-CO-CH ₃ , R ³ = pN (CH ₃ ) ₂ , R ⁴ = 0-C ₁₆ H ₃₃ and n = 2]

84 mg = 0.15 mmol of dried III [R ¹ = OH, R ³ = pN (CH ₃ ) ₂ , R ⁴ = O-C ₁₆ H ₃₃ and η = 2] are suspended in 190 mg = 2.4 mmol of dried pyridine. 122 mg = 1.2 mmol of acetic anhydride are added and the mixture is heated at 40 ° C. in a sealed flask for 6 hours. After this time, the thin-layer chromatographic control indicates complete conversion. Add 2 ml of water and heat for a further hour at 40 ° C. The reaction mixture is allowed to stand overnight and then lyophilized. The lyophilisate is reprecipitated from chlorofoim / acetone (1: 6). 90% of the pure end product is obtained as

white, highly hygroscopic powder.

Elemental Analysis: C ₃₀ H ₆₅ N ₂ O ₇ P · H ₂ O (604.8)

Ber .: N4,63P5,12

Gef .: N4.58 P5.06

Characterization of the PAF antagonistic effect

The antagonistic action of the compounds according to the invention (general formulas III and IV) is described below using the example of the inhibition of PAF-induced thrombocytogen aggregation. The latter was carried out in human platelet-rich citrated plasma by the method described by BORN (J. Physiol. (London) 162, 67-68 [1962]) using an ELVI 840 two-channel aggregometer (Elvi Logos, Milan, Italy) Compounds according to the invention exhibit a concentration-dependent inhibition of PAF-induced platelet aggregation, which can be reversed by increasing the PAF concentration SHIELD analysis of the observed parallel shift in the concentration-effect curve shows that the compounds according to the invention act as competitive antagonists of PAF quantitative measure of effectiveness are given in Table 1 the corresponding K _B values.

In contrast, induced by other triggers such as ADP, collagen or arachidonic acid platelet aggregation is influenced only insignificantly by the compounds of the invention.

Table 1:

Experimentally determined on human platelets Ke values

R 'R ² R ¹ R ⁴ rT K _B value

(Mol / I)

IiI 0-C ₁₆ H ₃₃ 0-C ₂ H ₅ PN (CH ₃ I ₂ III 0-C ₁₆ H ₃₃ OnC ₃ H ₇ PN (CH ₃ ), III 0-C ₁₂ H ₂₅ 0-C ₂ H ₅ pN (CH ₃ ) ₂ III 0-C ₁₆ H ₃₃ 0-C ₂ H ₅ P-NH ₂ III 0-C ₁₆ H ₃₃ OnC ₃ H ₇ P-NH ₂ III 0-C ₁₆ H ₃₃ 0-C ₂ H ₅ PN (CH ₃ I ₂ ! Il 0-C ₁₆ H ₃₃ 0-C ₂ H ₅ PN (CH ₃ I ₂ III 0-C ₁₆ H ₃₃ OnC ₃ H ₇ PN (CH ₃ I ₂ III 0-C ₁₆ H ₃₃ OnC ₃ H ₇ PN (CH ₃ I ₂

2 3,2- 10 " ⁶ 2 2,6-10 " ⁶ 2 > 10 " ⁵ 2 5,0-10 ⁶ 2 5.4 x 10 " ⁶ 3 4.4 to 10 " ' 4 3.6 to 10 " ' 4 6.3 to 10 " ' 5 1,1 -10 " ⁶

R '

R ⁴

Kn value (mol / l)

Ill III III III III III III

III III

III IV IV

0-C ₁₆ H ₃₃ 0-C ₁₆ H ₃₃ 0-C ₁₆ H ₃₃ 0-C ₁₆ H ₃₃

0-C ₁₆ H ₃₃

0-C ₁₆ H ₃₃ 0-C ₁₆ H ₃₃ 0-C ₁₆ H ₃₃

0-C ₁₆ H ₃₃

0-C ₁₆ H ₃₃

0-C ₂ H ₅ PN (CH ₃ ), 0-C ₂ H ₅ PN (CH ₃ ), nC ₃ H ₇ PN (CH ₃ J ₂ nC ₃ H ₇ PN (CH ₃ J ₂ Fi-C ₃ H ₇ PN (CH ₃ ), IV-C ₃ H ₇ PN (CH ₃ ), FV-C ₅ H ₁₁ PN (CH ₃ ) ₂ 0-CHR-C ₆ H ₅ PN (CH ₃ I ₂

0-C ₂ H ₅

0-C ₂ H ₅

0-C ₁₆ H ₃₃ OH OH 0-C ₁₆ H ₃₃ 0 -CH ₇ -C ₆ H ₅ 0-C ₁₆ H ₃₃ 0-C ₂ H ₅ 0-C ₁₆ H ₃₃ 0-C ₁₆ H ₃₃ - 0-CO-CH ₃ -

PN (CH ₃ I ₂ PN (CH ₃ J ₂ PN (CH ₃ J ₂ PN (CH ₃ J ₂ PN (CH ₃ ), PN (CH ₃ ), O-COCH 3 0-C ₁₆ H ₃₃

5 9.3-10 " ⁷ 6 1.9-10 " ⁶ 2 2.1 x 10 " ⁶ 3 9,6-10 " ⁷ 4 3.0-10 " ⁷ 8th > 10 " ⁵ 2 > 10 " ⁵ 2 9.2-10 " ⁶

3.5-10

2,8-10 "

> 10 " ⁶ >10" ⁵ > 10 " ⁵ 6.3-10" 3.9-10 "1.3-10"

Claims (4)

1. A process for preparing novel, highly active antagonists of the platelet-activated factor from the phospholipid series, characterized in that starting compounds of general formula I, ClI .., - - "R CII-R" (D o - P - occIi _n > ^ x
on where R 'is an unsubstituted, long-chain alkoxy radical having 12 to 20 carbon atoms, R ^{2 is} a methoxy, ethoxy, propoxy or benzoxy group or a methyl, ethyl or n-propyl to n-pentyl radical or a hydroxyl group, R ¹ and R ^{2 can} also be interchanged, for η are the numbers 2 to 10 and X is chlorine, bromine or iodine, with ortho, meta or para-substituted pyridines of the general formula II, (II) where R ^{3 represents} an amino group which may be substituted, identically or differently, by n-alkyl radicals having 1 to 4 carbon atoms, which together may also form a 3- to 7-membered ring, to give compounds of the general formula III, CIl _n -K " ¹ ' (III) where R ¹ , R ² and R ³ and η have the abovementioned meaning, and subsequently the compounds of general formula II, in the event that R ² is OH and R ¹ , R ³ and η are those indicated above have the meaning and R ¹ and R ² may be interchanged also tj with derivatives of short-chain carboxylic acids such as acetic acid or Propiniäure compounds of general formula IV, CII ... O-P - O-CCII where R ¹ , R ³ and η are as defined above and R ^{4 is} an acetyloxy or propionyloxy radical. 2. The method according to claim 1, characterized in that the reactions using anhydrous halogenated hydrocarbons, such as methylene chloride, chloroform, trichlorethylene or anhydrous l.ösungsmittelgemischen of chloroform / dimethylformamide or chloroform / acetonitrile (1: 1) are carried out with exclusion of moisture. 3. The method according to claim 1, characterized in that the reactions are carried out with a 2molaren excess of substituted pyridine and in a temperature range from 40 to 60 ⁰ C. 4. The method according to claim 1, characterized in that the reactions using anhydrous basic solvent, such as pyridine or triethylamine and with acid anhydride of the above-mentioned acids at 40 ° C are performed. Application of the invention The invention relates to a method for producing novel, highly effective antagonists of the platelet activating factor (PAF) with phospholipid structure. PAF is a naturally occurring phospholipid, which is a mediator of allergic and inflammatory reactions of great 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-asthmatic agents and antithrombotics.