DE3507721A1 - Process for the preparation of water-soluble C7-hydroxylated cholesterols and desmosterols - Google Patents

Abstract

The present invention relates to a process for the preparation of water-soluble derivatives of 7 beta -hydroxycholesterol ( DELTA <5>-cholestene-3 beta ,7 beta -diol) and of 7 beta -hydroxydesmosterol ( DELTA <5,24>-cholestene-3 beta ,7 beta -diol) and related steroids, which is characterised in that the corresponding steroid diols are reacted with succinic anhydride and pyridine to give the corresponding bis-hemisuccinates and the latter are converted into salts of alkali metals and/or biogenic amines.

Description

The invention relates to a method for the preparation of water-soluble CL-hydroxylated cholesterols and desmcsterols.

There are many uses to make steroids water-soluble generally the reaction of the same with succinic anhydride and either forms the resulting hemisuccinate Sodium or potassium salt.

In the case of Tß-hydroxy-cholesteryl-bishemi-

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succinates and its Δ-cholesteric-Sβ, Tβ-di-Na-bishemisuccinate or the 7β-hydroxy-desmosteryl-bishemisuccinate and its Δ'-cholesteryl-Bβ, Tβ-di-Na-bishemisuccinate turn out to be both the di-sodium and the di-potassium salt are relatively poorly soluble in water; the solutions are cloudy and only in ■ high dilution clear at room temperature.

Another disadvantage of the salts of these two alkali metals is that they can be dissolved without significant loss of substance do not allow recrystallization from water. Manufactured in ethanol, they cannot be recrystallized from this solvent either, but can only be washed on the suction filter with ethanol, which creates a possible slight excess the sodium ethylate used for their production can only be removed completely with difficulty.

It has now been found that the corresponding salts of the steroid carboxylic acids of the above type with colamine (fithanolamine) have a significantly increased solubility in water compared to the corresponding sodium or potassium salts. For example, give 1000 mg in 500 ml of water is still clear at 2O ⁰ C solution, which

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is of importance for the dosage as an anti-cancer agent.

From a biochemical point of view, the biogenic amine colajnine is im Organism metabolized by decarboxylation of the amino acid serine. It is part of the phosphatides and at the same time the parent substance of choline, its acetylation product as Action substance of the cholinergic nerves at the synapses or motor end plates of the nerve endings as a neurotransmitter is effective.

The succinate plays in the citric acid cycle as succinyl-CoA (Coenzyme A) plays an important role (see textbooks on biochemistry).

Thus, the constituents of the water-soluble molecules are 7ß-Hydroxy-cholesterol or -desmosterols, as well as these themselves, to be regarded as endogenous biochemical substances, their Substitution exerts an additional beneficial influence on the cancer patient's organism. By now in water-soluble Form the body's own anti-setting agent is present their effect increased.

The invention becomes insane. the following based on exemplary embodiments further explained:

Example I.

Representation of Tß-hydroxy-cholesteryl-bishemisuccinate.

(Δ-cholesterol-Sß, Tß-di-bis-hemisuccinate)

a n

SmD. 12O ⁰ C 602, SI

Srrp. 178.5 ° C Snp. 159.0 ⁰ C

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In a 1 liter three-necked flask equipped with a stirrer, reflux condenser, thermometer and gas line pipe, MO g (= 0.1 mol) of 7β-hydroxycholesterol, 40 g (= 0.4 mol) of Bernsteir acid anhydride and 60 ml (= 0.75 mole) of pyridine under a N -Atnr sphere unt --- r stirring slowly melted u'.d five (5) Stunder, at 70 to 75 ⁰ C internal temperature kept.

After cooling to approximately 3O ⁰ C, 500 ml of distilled H _£ 0 were added with stirring. The reaction product initially settles as a highly viscous, viscous mass that gradually solidifies. The stirrer was switched off, the supernatant aqueous pruning was poured off and the solid white cake was washed thoroughly three times with nestled water. This was then poured over 800 ml of CH OH (methanol) in the flask and brought into solution by heating with stirring on a reflux condenser.

The yellow methanolic solution was then irr ^{at -1 0 C.} Leave the refrigerator to crystallize. After the suction device, on a G4 suction filter, residual pyridine or its salts were removed by thorough washing with water. In the process, some substance is lost, which can be observed in the initial turbidity of the washing water in the feeding bottle. It is washed se with distilled H ₂ O until the "aufende wash water at is clear. After three hours of drying in an oven at 8O ⁰ C 31.1 g (= 51.56% of theory) of crude product. M.p. . 159, O ⁰ C.

The mother liquor was concentrated to half (approx. 400 ml) and for 12 hours at -18 ⁰ C in the freezer of the crystallization

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left.

Yield: 4.55 g (7.56% of theory); Srnp. 159 ⁰ C.

The filtrate of this fraction (about 400 ml) was then diluted with 40 ml H ₂ O and again 12 hours auroewahrt at -18 ⁰ C. After suctioning off and drying, a further 2.5 g (= 4.15% of theory) of white crystals with a melting point of 159 ^° C. were obtained. This total crude yield was thus 63.27% of theory.

The total amount of all the three fractions was last suspended in distilled water and washed by means of a turbo mixer again several times, filtered with suction, and finally dried at 8O ⁰ C overnight. The yield of pure product was then a total of 37.85 g (= 62.87% of theory); M.p. 159, O ⁰ C.

The accompanying Figure 1 shows the infrared spectrum according to Example 1 prepared compound 7β-hydroxy-cholesterylbishemisuccinate.

Example II

Representation of the di-sodium salt of 7ß-hydroxy-cholesterylbishemisuccinate (Δ-Cholester-3β, 7β-disodium bis-hemisuccinate).

ZC _z H _s OhJg. 136.114

° 602.81 o ^ r ^ ^H 92.7 36 ° 646.788 © - ^^ © M «-

Srrp. 159.0 ⁰ C ° m.p.> 360 ^c C »

BATH ORIGINAL

6.0 g (0.01 mol) of Δ-cholesterol-Sß, 7ß-bishemisuccinate were dissolved in 60 ml of 95% CpH-OH (ethanol) and mixed with the solution of 0.4578 g of metallic sodium in 60 ml of 95% igtm C ₂ H OH (ethanol) added with stirring. The di-sodium salt is currently separating out. After the suction on a G4-suction filter and washing with a little cold 95% CpHf-OH (ethanol) was dried in an oven at 8O ⁰ C.

Yield: 6.45 g (^ quantitative); Mp.> 36O ⁰ C.

The substance is readily soluble in water, but with the restriction that it can be used over a long period of time down to very low concentrations Standing at room temperature in the solution becomes cloudy.

The accompanying Figure 2 shows the infrared spectrum according to Example 2 prepared compound of the di-sodium salt of 7ß-hydroxy-cholesteryl-bishemisuccinate.

Example III

Representation of the di-ethanolamine salt of 7ß-hydroxy-cholesterylbishemisuccinate (Δ-cholesterol-Bß, 7ß-di-ethanolamine-bishernisucc: nat)

Srrp. 159.0 ⁰ C

BATH

In a 250 ml three-necked flask with stirrer, dropping funnel and reflux condenser, 6.0 g (0.01 mol) of the steroid dicarboxylic acid in 100 ml of C _? Dissolved H OH (ethanol) while stirring from the dropping funnel with a solution of 1.23 g (0.02 mol) of colamine (aminoethyl alcohol) in 20 ml of C_H OH. The colorless at the boiling temperature of the ethanol clear solution was left after transferring into an Erlenmeyer flask for several hours at -1 ⁰ C in the refrigerator for crystallization. After filtration with suction on a G ¹ I-FiIternutsche, washing with ice-cold ethanol and drying at 8O ⁰ C in an oven were obtained 7.2 m g (= quantitative) of the water-soluble salt. M.p. 168 to 169 ^° C.

A solution of the same, prepared by dissolving 1000 mg (= 1.0 g) of the substance in 500 ml of distilled water, corresponding to a concentration of 1.0 mg in 0.5 ml of water, remained in the refrigerator even when it was stored for 24 hours completely clear at -1 ^{0 C.}

The accompanying FIG. 3 shows the infrared spectrum of the compound of the di-ethanolamine salt prepared according to Example 3 of Yβ-hydroxy-cholesteryl-bishemisuccinate.

Claims (1)

Expectations f 1. J Process for the preparation of water-soluble derivatives of Tß-kydroxycholesterol (Δ-cholesterol ~ 3ß, 7ß-diol) and 7ß-hydroxy-desmosterol (A ⁵ '-cholestadiene-Sß, 7ß-diol) and related steroids,
characterized, that using the appropriate steroid diols succinic anhydride and pyridine are converted to the corresponding bis-heraisuccinates and these are converted into salts transferred from alkali metals and / or biogenic amines. 2. The method according to claim 1, characterized, ' ., that the implementation of one mole of these steroid diols with ί at least 4 moles of succinic anhydride has to be made. «R 3 process according to claims 1 and 2,
characterized, that the salts of the steroid dicarboxylic acids which can be prepared in this way not only be produced in aqueous solution by means of the hydroxides of the alkali metals, but advantageously by dissolving the bishemisuccinates of these steroids in Ethanol and subsequent addition of the corresponding alcoholate of the alkali metals. H. The method according to claims 1 to 3>
characterized, that the salts of the steroid dicarboxylic acids of alkali metals thus obtained are water-soluble. BATH 5. The method according to claims 1 to 3,
characterized, that for the production of water-soluble derivatives of the steroid derivatives according to claim 1, bicgenic amines can also be used instead of the salts of alkali metals. 6. The method according to claims 1 to 5,
characterized, that the production of such water-soluble salts of
Steroid dicarboxylic acids according to claim 1 in ethanol solution using the biogenic amine colamin
(Aminoethyl alcohol) and other biogenic amines. 7. Use of the salts of bis-hemisuccinates of 7ß-hydroxy-cholesterol represented according to claims 1 to 6 (Δ-cholesterol-S [beta], 7 [beta] -diol) and / or of 7 [beta] -kydroxydesmosterol ([Delta] 1 cholesterol-S [beta], 7 [beta] -diol) and
related steroids as native, biochemical and therefore harmless signal substances that are universal in the warm-blooded animal kingdom for modulating the immune word in cancer of all phenotypes. BAD Cfl! G! NAL