DE1668809B2 - Prostaglandin compounds, processes for their preparation and therapeutic agents containing them - Google Patents

Description

in which R is a hydrogen, alkali or alkaline earth atom or the ammonium radical or an alkyl group Means 1 to 5 carbon atoms and which is the stereochemical configuration of naturally occurring Have prostaglandin.

2. Process for the preparation of the prostaglandin compounds according to claim 1, characterized in that that you have an all-cis-triene compound of the general formula

CH ₃ - (CH ₂ ) ₅ - (CH = CH - CH ₂ ) ₃ - (CH ₂ ), - COOM in which M denotes a hydrogen, alkali or alkaline earth atom or the ammonium residue, with an enzyme system suitable for prostaglandin synthesis incubated under aerobic conditions in a manner known per se and optionally then converted the reaction product obtained into an alkali, alkaline earth or ammonium salt or an alkyl ester having 1 to 5 carbon atoms in the alkyl group in a conventional manner.

3. Therapeutic agent, characterized by a content of a compound according to claim 1 in addition to the usual inert and pharmacologically acceptable carriers.

The invention relates to prostaglandin compounds, processes for their preparation and them containing therapeutic agents.

A number of related substances known as prostaglandins and found in humans and other mammals have been found to display a wide range of pharmacological activities. These substances all have structures based on those of prostanoic acid (PA), the 1 <x- (6-carboxyhexyl) -5 /? - octylcyclopentane and has the following structural formula:

CH ₂ CH- (CH ₂ J ₆ -COOH

CIl ₂ CH- (CH ₂ J ₇ -CH.,
CH,

A typical prostaglandin is Prostaglandin E, commonly referred to as PGEi, which is the following Structural formula has:

O = C CH- (CH ₂ ) "- COOH

I I

CH ₁ CH-CH = CH-CHOH- (CH,) ₄ -CH,

\ /
CH

OH

The mentioned prostaglandin egg contains two hydroxyl groups, an oxo group and a carboxyl group. It is assumed that PGEi the following has stereochemical configuration (Ha)

'• ■ \ / \ / \ / COÖH

(Ha)

OH

The structural relationship between the prostaglandins and fatty acids suggests that the latter Were precursors to prostaglandins, and leads e.g. B. on the biochemical synthesis of PGEi from bis-homo -}> linolenic acid (Eicosa-8c, llc, 14c-trienoic acid), described YQrj Von Qnrη Bserthuis, N lower, and Vonkeman, Biochem. Biophys. Acta, 1964, 90, 204; Nature 1964, 203,839, and also by Bergström and Employees, using enzymes present in sheep vesicular glands. The acid precursors are characterized in that one has a double bond system in which three double bonds through individual methylene groups are separated.

Since the pharmacological activities of natural prostaglandins are various, including an effect on smooth muscles, on the cardiovascular system, on the central nervous system

bo and in terms of the mobilization of lipids Attempts have been made to produce prostaglandins from acids with more and less than 20 carbon atoms to study their properties. In many cases, the use of acids failed with the above double bond systems the enzymatic synthesis. Among the variants tried are synthetic acids with an odd number of carbon atoms in the molecule (natural fatty acid

ren have an even number of carbon atoms). These can be referred to as unnatural prostaglandins will. Two such acids that have been successfully converted to members of the prostaglandin series, are generally heneicose-9,12,15-trienoic acid having the cis configuration and Nonadeca-T.lO.lS-trienoic acid, each of the above double bond system, the one at the sixth carbon atom, from the terminal In terms of the methyl group, it starts like this at the natural precursor acids is the case, have and which differs from the bis-homo-y-ionoic acid thereby distinguish that they have one more methylene group and one less on the one carrying a carboxyl group Have side chains, and are therefore referred to as α-homo- and Λ-noric acids. They are called Prostaglandins "-Homo-PGEi and" -Nor-PGEi and have the following structural formulas IV and V:

O = C CH- (CH ₂ J ₇ -COOH

CH, CH-CH = CH-CHOH- (CH ₇ U-CHj

\ /
CH

OH
O = C CH- (CH ₂ I ₅ -COOH

CH, CH-CH = CH-CHOH- (CH ₂ U-CH ₃

\ /
CH (IV) («-Homo- PGEi)

(V) (A-Nor-PGE,)

OH

The pharmacological activities of prostaglandins are specific and have many compounds not the properties that their closest homologues show. jo

Not only has a class of triene compounds having a cis configuration been found that can be converted to prostaglandins by enzyme synthesis, but are

CH., - (CH ₂ ) ₅ - (CH = CH-C

in which M represents a hydrogen atom or an alkali or alkaline earth atom or the ammonium radical. Under the compounds corresponding to this formula fall Heneicosa-8c, l 1c, 14c-trienoic acid (the designations "C" and "t" denote the cis and trans configurations of the double bond at a given point), which have the above double bond system and thereby differ from bis-homo-y-linolenic acid. thus products and their derivatives have also become available which have a strong pharmacological activity and in many cases even better effects compared to the natural prostaglandins and own other synthetic prostaglandins. These triene compounds, which generally have a cis configuration have the general formula

, - (CH ₂ ) ₅ -COOM

that they have one more methylene group on the side of the double bond system that of the carboxylic acid group away. They are therefore derived from the ω-homoacid.

These triene compounds are made by incubation with a prostaglandin synthesis enzyme system the following compounds of the general formula III under aerobic conditions

O = C CH (CH ₂₎ ,, - COOR

CH ₂ CH- CH = CH- CHOH- (CH,) ₅ -CH,

\ /
CH

OH (III)

educated. In this general formula III, R has the meaning of M or is 1 to 5 carbon atoms containing alkyl group.

It is believed that the compounds of this invention as derivatives of 4ot-hydroxycyclopentan-2-ones, those in the 1'-position a 6-carboxyhexyl or 6-alkoxycarbonylhexyl group and in the 5j3-position a 3 S-hydroxyalkyl-lt-enyl group with 9 Carbon atoms (where S is the designation of the stereochemical configuration in the Cahn-Ingold-Prelog system is). ω-HomoPGEi can also be used as l (X- (6-carboxyhexyl) -4a-hydroxy-5 /? - (3 S-hydroxynonlt-enyl) -cyclopentan-2-one (purple, R = H). The configuration of this connection is as follows:

bO OH

'/ \ / \ / \ / COOR

OH

(MIa)

(. »- Homo-PG E ₁ ) R = H

The compounds of the general formula III, in which R has the above meaning, can thereby

be produced that one in a known manner an all-cis-triene compound under aerobic conditions the general formula

CH ₃ - (CH ₂ ) ₅ - (CH = CH-CH ₂ J ₃ - (CH ₂ I ₅ COOM

in which M has the preceding meaning, with a to Prostaglandin synthesis suitable enzyme, e.g. B. shredded sheep vesicular glands, incubated and possibly then the reaction product obtained in the usual way in an alkali, alkaline earth or ammonium salt or an alkyl ester having 1-5 carbon atoms is preferably converted to the alkyl group Glutathione added as a cofactor. Furthermore, the addition of an antioxidant, e.g. B. hydroquinone, cheap.

After the enzymatic conversion, the reaction product is converted into a suitable derivative The free acid can be isolated from the reaction mixture and used as such, or it can in the usual way in a salt, ζ. Β. their sodium, potassium, ammonium or calcium salt, or into an ester of one lower alcohol with 1 to 5 carbon atoms ζ. Β. the methyl or ethyl ester.

Such esters can be made from the corresponding alkanol in the presence of a known catalyst or with Help an essential solution, ζ. Β. of diazomethane or diazoethane.

The starting acid can be obtained by selective partial hydrogenation of the corresponding triic acid, which itself obtainable by the use of classical synthetic methods, or by chain extension of the corresponding trienoic acid with one carbon atom less using the Arndt-Eistert reaction getting produced. Both triic acid and Heneicosa-8c, l lc, 14c-trienoic acid are new compounds.

The prostaglandins according to the invention exhibit smooth muscle contraction activity, hypotensive or hypertensive activity, cardiac stimulating activity, and activity in inhibiting platelet accumulation. The following table summarizes the potencies obtained for ω-Ήοπιο-PCEi compared to their close homologues, a-Homo-PGEi, a-Nor-PGEi, α-ΝθΓ-ω-Homo-PGEi, a-Bisnor-PGE, and Λ-Bishomo-PGEi were found, which are given in the structural formulas IV and V and the structural formulas Vl to VIII below.

O = C

CH -

I I

CH ₂ CH-CH = CH-CHOH- (CH ₂ U-CHj

\ /
CH

OH

(VIl
(■ »-Bishonio-PUE,)

O = C CH- (CH ₂ I ₄ COOH

CH ₂ CH-CH = CH-CHOH- (CH ₂ U-Ch,

\ /
CH

OH

O = C CH- (CH ₂ I ₅ -COOH

CH ₂ CH-CH = CH-CHOH- (CH ₂ ) ₅ -CH ₃

\ /
CH

OH

(Viii

(\ -Bisnor-PGE |)

(VIII)
-PGEi)

The substances were in the form of a solution of the sodium salt or the methyl ester in a physiological Saline solution compared to PGEi tested as a control sample.

Smooth muscle contraction activity (A) was demonstrated by acting on the isolated guinea pig ileum (Guinea pig ileum) using the method of Magnus (cf. E. W. Horton et al., Brit. J. Pharmacol. Chemotherapist 21, 1963, 182-189). The lowering of blood pressure (B) was measured using Wistar rats at a dose of 3 μg / kg body weight. (See D. W. du C h a r m e et al. Prostaglandins, Proc. 2nd Nobel Symposium Stockholm, June 1966, edited by S. Beig si rue in and B. Sammeisson.

Pp. 173-182, Interschience Publishers, New York, 1967. Cardiac stimulus activity was measured by perfusion of an isolated rat heart using the Langendorff method modified by suspending the heart and corona flow rate (Ci) bo simultaneously was measured with the contraction force (C ₂ ) at dosage levels in μΐη / ηιΙ (Cc) (cf. F. Bt: -ti et al., Med. Pharmacol. Exp. 13, 233-240 (1965). The inhibition of platelet accumulations (D ) was measured in vitro using rat blood according to b5 of the Born and O'Brien method at doses of 0.5 to 1 μg / ml (cf. GV Born, Nature 194, 927 [1962] and JR O'Brien η , J. CI i η. Pathoi. I 5.446 ii 9621).

link

Λ Β

Cc

Cl

C2

ω-homo-PGE, (III, R = H) 120-130

PGE, (II) 100

ff-Homo-PGE, (IV) 2

ff-Nor-PGE, (V) 1

a-Nor-w-homo-PGE, (VIII) 2

ff-Bisnor-PGE, (VII)
a-Bishomo-PGEi (VI)
PGEi methyl ester
w-Homo-PGEi-Methylestercrdll, R = CH.,)

60-70 0.3 200 100-200 400-500 70 1 200 50-200 100 1 100 100 1 1 100 100 0 1 100 100 1 1 120 100 2 1 100 100 1 125-175 200

It can be seen that ω-homo-PGEi is 4 to 5 times so effective as PGEi as a platelet aggregation inhibitor and at least 3 times as effective as a coronavasodilator is.

The ability to inhibit platelet accumulation or aggregation is very noteworthy since it is known that such aggregation can result in thrombosis and perhaps also atherosclerosis. ω-Homo-PGEi also inhibits platelet aggregation induced by adenosine diphosphate in vivo, z. B. in rats.

The invention also relates to therapeutic agents, which by containing a compound of the general Formula III are characterized in addition to the usual inert and pharmacologically acceptable carriers. The carrier material can consist of a solid or a liquid in which the compound is dissolved, is dispersed or suspended. The preparations are preferably in dosage units and in Form of tablets, powders, capsules, solutions, emulsions and liquid suspensions, especially in injectable form.

The invention is explained in more detail below with reference to examples in which all temperatures are given in ⁰ C and TLC, RPPC and GLC mean thin-layer chromatography, reversed-phase partition chromatography or gas-liquid chromatography.

example 1
A) Preparation of the starting material

a) Heneicosa-8c, Hc, 14c-trienoic acid through
Arndt-Eistert synthesis

127 mg (1 mmol) of oxalyl chloride were added to 127 mg (0.4 ImMoI) of Eicosa-7c, 10c, 13c-trienoic acid (Struyk et al., Rec.trav.chim., 1966, 85, 1233) in 0.3 ml of benzene; the mixture was allowed to stand at room temperature for 4 hours, after which the benzene and excess oxalyl chloride were removed by a stream of nitrogen. The remaining Eicosa-7c, 10c, 13c-trienoyl chloride was dissolved in 0.4 ml of benzene and the solution was added dropwise to 5 ml of a shaken 0.4 η solution of diazomethane in ether at 0 ^° C. After standing at 0 ° C. for 30 minutes, excess diazomethane, benzene and ether were removed by a stream of nitrogen. The remaining diazoketone was dissolved in 1 ml of anhydrous methanol and 44 mg of silver benzoate in 0.55 ml of triethylamine were added over the course of 90 minutes. The mixture was then heated to boiling for 2 minutes.

It was then filtered and the residue was washed with 1 ml of ether. The combined filtrates were used for Evaporated dry. The product was determined by TLC following the procedure of Morris, Chem. And Ind., 1962, 1238, using two plates from

2 ", silicon dioxide gel G with a thickness of 1 mm, containing 26% silver nitrate, based on the dry silicon dioxide, were impregnated and a mixture of benzene and ether (90:10 vol.) as the eluent purified, whereby 70 mg methylheneicosa-

jo 8c, l lc, 14c-trienoate in Fo ^r m yielded an oil of 99% purity, as can be found from GLC analysis. This ester was saponified by treatment with 2 ml of 0.5 M aqueous methanolic potassium hydroxide at 70 ° C within 1 hour under nitrogen, with potassium hydrogen

j-, neicosa-8c, llc, 14c-trienoate resulted, of which the free Acid by adding dilute sulfuric acid until a mixture with a pH of 3 is obtained was released, with the organic acid extracted with light petroleum, the solvent

4 (i evaporated and the free acid in the form of an oil was left behind.

b) Manufacture of

Heneicosa-8c, l lc, 14c-trienoic acid and their
5 methyl ester by hydrogenation of

Heneicosa-8,11,14-triic acid

1.) Production of Heneicosa-8,11,14-triic acid

To a solution of 30.8 g (0.2 mole) of non-8-ynoic acid (n = 1.4529%, mp 20-2 ⁰ C in 100 m! Of tetrahydrofuran were added 275 ml of 1.45 η solution Äthylmagnesiumbromid in tetrahydrofuran was added within 30 minutes at 0 ⁰ C. the obtained mixture was stirred for 2V2 hours, after which 720 mg (4 mmol) of copper (I) cyanide were added and stirring was continued for 10 minutes. A solution of 36.2 g (0.15 mol) l-bromododeca-2,5-diyne (Struyk et al., Loc cit, bp 78-80 ° C / 0.001 mm Hg, n | ⁵ = 1.5055) in 75 ml of tetrahydrofuran was then added within 10 Minutes

ω added, the stirred mixture heated to the boil and refluxed for 15 hours; at the end of the first 4 hours another subset of Copper (I) cyanide (360 mg) added. After the reflux treatment was finished, the main

b5 amount of solvent present under reduced Pressure distilled off 100 ml of ether and aqueous were added to the cooled residue 2 η-sulfuric acid added. The mix was

shaken thoroughly, the ether layer which formed on standing was separated and the aqueous layer extracted with ether. The combined ether layers were washed with saturated aqueous ammonium chloride solution and then with water, then dried with anhydrous sodium sulfate and evaporated. The residue was dissolved in a small amount of light petroleum and the solution was chromatographed on a column (30 cm in length and 2.2 cm in diameter) of Siliciumdioxydgei, using mixtures of light petroleum and ether, which started a volume ratio of 95: 5 and were used as the eluent continued with increasing proportions of ether up to a ratio of 40:60. The eluted fractions which were found to contain acid were pooled and evaporated; the residue was crystallized from a mixture of light petroleum and ether (85:15 by volume) at -5 ° C. and gave 22.7 g (48%) of heneicosa-8,11,14-triic acid, melting point 56 to 57.5 ° C, infrared absorption peaks at 1690 (-COOH) and 1309cm- '(-CH ₂ -C = C-). Nuclear magnetic resonance spectrum (NMR, the relative intensity and the designation are given in brackets):

Distorted triplet at <5 0.90 (3; CH ₃ -),

Complex at 1.1 to 1.85 (16; -CH ₂ -),

Complex at <51.85 to 2.5 and

a triplet at ό 2.32
(6; -CH ₂ -C = C- and -CH ₂ -COO-),

Complex at <5 3.03 (4; -C = C-CH ₂ -C = C-)
and

a broad singlet (l; -COOH).
The acid was converted into its methyl ester by reaction with an ether solution of diazomethane, which was subjected to GLC and showed a purity of 99.5%; Mass spectrum: peak from the molecular ion, derived at m / e = 328.

2.) Manufacture of

Heneicosa-8c, 11c, 14c-trienoic acid

and their methyl esters

100 mg Lindlar catalyst and 0.05 ml of quinoline were added to a solution of 674 mg (2.15 mmol) Heneicosa-8, ll, 14-triinsäure in 30 ml of methanol, and the mixture was shaken in hydrogen at 0 ⁰ C until the calculated volume of hydrogen to convert the triple to a double bond had been absorbed. The solution was filtered and poured into a mixture of 50 ml of water and 3 ml of sulfuric acid. The mixture obtained was extracted with 3 parts of optically pure light petroleum, the extract was washed with water and dried with anhydrous sodium sulfate, and the solvent was removed by evaporation, 677 mg (98%) of crude Heneicosa-8c, IIc, 14c-trienoic acid remaining as residue ; π J ⁵ = 1.4765; GLC of the methyl ester showed the presence of 1% monoic, 9% diene and 90% trienoic acid. The acid (246 mg, 0.77 mmol) was converted to its methyl ester by the addition of an ethereal solution of diazomethane and the ester was determined by TLC according to the method of Morris Lc. using plates of silica gel with a thickness of 1 mm, which were impregnated with 26% silver nitrate based on the weight of the dry silica, and a mixture of benzene and ether (volume ratio 5: 1) as eluent, with 174 mg ; (68%) gave methyl heneicosa-8c, lc, 14c-trienoate as an oil of 99.5% purity as indicated by GLC; n '" ⁵ = 1.4695; Infrared absorption peaks at 3000, 1400, 720 (cis-CH = CH-), 1743cm- '(-COOCH ₃ ); Mass spectrum: tip from the molecular ion, derived at m / e 334, last double bond between carbon atoms 14 and 15; NMR spectrum:
distorted triplet at δ 0.89 (3; CH ₃ -),
Complex at όΐ, ΐ - 1.8 (16; -CH ₂ -),
Complex at δ 1.8 - 2.4 and
ίο triplet at (5 2.20

(6; -CH ₂ -CH = CH- and -CH ₂ -COO-),
Complex at ö 2.75

(4; -CH = CH-CH ₂ -CH = CH-),
Singlet at 3.57 ö (3; -COOCH _3), and
i-> complex at 0 5.29 (6; -CH = CH-).

The methyl ester (150 mg, 0.45 mmol) was obtained by heating with 20 ml of a 0.35 η solution of sodium hydroxide hydrolyzed in 85% methanol at 60 ° C within one hour under nitrogen. The reaction mixture was poured into 100 ml of water, with

4 η-sulfuric acid acidified to a pH of 3 and extracted with ether. The essential extract was dried with anhydrous sodium sulfate and evaporated, leaving almost pure Heneicosaec.llcHc-trienoic acid in the form of an oil; n " = 1.4788, with a content according to its ultraviolet spectrum of 5% conjugated diene; infrared absorption peaks at 1715 and 3010 cm- '; mass spectrum: peak derived from the molecular ion at m / e 320,

jo double bond between carbon atoms 14 and 15; Double bonds between the carbon atoms 8 and 9, and 14 and 15 according to analysis by means of ozonolysis.

B) invention

J5 A mixture of 24 mg Heneicosa-ec ,! lc, 14c-trienoic acid, 120 mg glutathione, 12 mg hydroquinone and 600 mg protein from the particulate enzyme fraction of sheep vesicular glands (obtained as from

5 truyk et al. Loc. cit. described), suspended in aqueous 0.2 m-tris- (hydroxymethyl) -aminomcthan-hydrochloride buffer solution, pH 8.0, was aerobic to a total volume of 300 ml by stirring the mixture in a large container open to the air incubated at 30 ⁰ C within 20 minutes.

The enzyme reaction was then terminated by acidifying with citric acid to a pH of 4, and the mixture was extracted with 2 aliquots of 300 ml of ether. The residue from evaporation of the extract was dissolved in chloroform and the solution was chromatographed on a column of silica gel (25 g), using successively 150 ml of chloroform, 150 ml of chloroform with a content of 3% methanol and 150 ml of chloroform with a content of 5% as the eluent. Methanol were used. 10 ml fractions of eluate were collected. Small samples of the fractions (0.1 ml) were checked for the presence of a prostaglandin by treatment with alkali (0.5 ml of 3N KOH in 75% methanol) and examination for an ultraviolet absorption peak at 278 nm The last fractions of the eluent with 3% methanol and the first fractions of the eluent with 5% methanol were found to contain the desired substance; they were evaporated to give 12 mg of product. This product was then further purified by RPPC using a diatomaceous earth (23 g) made water repellent by treatment with methylchlorosilane (mixture) as a carrier, a

Mixture (1.8 ml) of equal volumes of isooctanol and chloroform as the stationary phase and aqueous methanol containing 9 volumes of methanol to 10 volumes of water as the eluent. All those fractions (2 ml each) which showed the alkali reaction described above, with the exception of the first and the last, were combined and evaporated to give 7.2 mg of 1 a- (6-carboxyhexyl) -4 <x-

hydroxy-5 jSJ- (3 S-hydroxynon-1t-enyl) -cyclopentan-2-one (ω-homo-PGEi) in the form of a crystalline substance, mp 115 ° C to 116 ° C (from light petroleum / ether). A comparison with PGE, by TLC on silica gel using a mixture of chloroform. Methanol. Acetic acid and water in the corresponding proportions by volume of 90: 8: 1: 0.75 gave an Rf value for ω-homo-PGEi of 0.34 compared with 0.32 for PGEi, the two substances being just separated. The rotation, measured in a 1 g / 1 solution in methanol at 20 ⁰ C was -69 ° C.

By treatment with 0.5 η-alkali at 20 ° C within of 10 minutes the ω-homo-PGEi (I mg) was obtained as described above to an acid dehydrated, which showed an ultraviolet absorption maximum at 278 nm. The product was in his Converted methyl ester by reaction with an ethereal solution of diazometnane.

The mass spectrum of the methyl ester had a molecular ion peak at m / e = 364 and a reference peak with maximum intensity at m / e = 219. After conversion of the methyl ester into its trimethylsilyl ether, it appeared to be homogeneous (by GLC at 200 ^° C. using a diatomaceous earth, which was loaded with 1% polyethylene glycol adipate was found to be the carrier

ίο a retention time of 1.25 compared to that of PGEi = 1.0).

Example 2

3 mgü) -Homo-PGE | were dissolved in 1 ml of ether, and an excess of an ethereal solution of diazomethane at 0 ^° C. was added. After 5 minutes, the mixture was evaporated to dryness by a stream of nitrogen, whereby the methyl ester of ω-homo-PGEi (formula III, R = CFh) was obtained in the form of an oil. Its Rf value is 1.05 compared to that of PGEi methyl ester (1.00) in TCL over silica gel; the eluent consisted of a mixture of chloroform and methanol in a volume ratio of 96: 4.

Claims (1)

Claims: 1. Prostaglandin compounds of the general formula O = C CH- (CH ₂ J ₁₁ -COOR ₂ CH-CH = CH-CHOH - (CH ₂ I ₅ -CH ₃ CH OH