DE1518793C

DE1518793C - Process for the production of alkali salts of phosphocreatine excretion from 1260472

Info

Publication number: DE1518793C
Application number: DE19591518793
Authority: DE
Inventors: Jesus Paris Anatol
Original assignee: Fa Ugine Kuhlmann, Paris
Priority date: 1958-09-26
Filing date: 1959-09-24
Publication date: 1973-04-26

Description

mit 50%igem Alkohol, worauf man im Ofen bei 50° C trocknet. Man erhält auf diese Weise 132 Teile Dibenzyloxyphosphorylkreatinin mit einem Schmelzpunkt von 91 bis 92⁰C, also eine Ausbeute von 71%.with 50% alcohol, after which it is dried in the oven at 50 ° C. In this way, 132 parts of dibenzyloxyphosphoryl creatinine with a melting point of 91 to 92 ^° C., that is to say a yield of 71%, are obtained.

Dieses Dibenzyloxyphosphorylkreatinin eignet sich ohne weitere Reinigung sehr gut zur Herstellung von Phosphokreatinin und schließlich Phosphokreatin.This dibenzyloxyphosphoryl creatinine is very suitable for the production of Phosphocreatinine and finally phosphocreatine.

Zur Analyse kristallisiert man das Produkt aus 70%igem Methanol um, wobei sich die Ausbeute auf 90% und der Schmelzpunkt auf 93 ⁰C beläuft.For analysis to the product crystallized from 70% methanol in order, wherein the yield to 90% and the melting point to 93 ⁰ C amounts.

Analyse für C₁₈H₂₀N₃C₄P:Analysis for C ₁₈ H ₂₀ N ₃ C ₄ P:

Berechnet ... C 57,90, H 5,40, N 11,25, P 8,30%; gefunden ... C 57,68, H 5,96, N 10,89, P 8,54%.Calculated ... C 57.90, H 5.40, N 11.25, P 8.30%; Found ... C 57.68, H 5.96, N 10.89, P 8.54%.

186,5 Teile Dibenzyloxyphosporylkreatinin werden aufgelöst in einem Gemisch aus 500 Raumteilen einer 2n-Natronlauge und 150 Raumteilen Alkohol. Dieser Lösung setzt man Palladiumschwarz hinzu (hergestellt durch Hydrieren eines Gemisches aus 95 Teilen Ruß, 1000 Teilen Wasser und 9,3 Teilen Palladiumchlorid), worauf man bei gewöhnlicher Temperatur und normalen Druck hydriert. 20 500 Raumteile Wasserstoff werden in 2 Stunden absorbiert.186.5 parts of dibenzyloxyphosporylcreatinine are dissolved in a mixture of 500 parts by volume of a 2N sodium hydroxide solution and 150 parts by volume of alcohol. Palladium black is added to this solution (prepared by hydrogenating a mixture of 95 parts of carbon black, 1000 parts of water and 9.3 parts of palladium chloride), whereupon hydrogenation is carried out at ordinary temperature and pressure. 20 500 parts by volume of hydrogen are absorbed in 2 hours.

b) Verfahren der Erfindungb) method of the invention

Für die Spaltung der Alkalisalze des Phosphokreatinins filtriert man die Reaktionsmasse zur Entfernung des Katalysators und wäscht dann den Filterrückstand mit 250 + 100 + 100 Teilen Wasser. Dem Filtrat setzt man 47,4 Teile Natriumhydroxyd, aufgelöst in 85 Teilen Wasser, hinzu. Man erhitzt in 10 Minuten auf 80° C. Die Lösung wird anschließend auf etwa 10°C gekühlt; man stellt deren pH-Wert in Gegenwart von Phenolphthalein mit 1182 Raumteilen η-Salzsäure auf 7,8 ein, filtriert und setzt der Lösung 9480 Teile Alkohol hinzu. Das Dinatriumsalz des Phosphokreatins scheidet sich in Form eines Öls ab. Man läßt es zum Auskristallisieren 2 oder 3 Tage bei Raumtemperatur stehen, filtriert, wäscht mit 80%igem Alkohol, dann mit absolutem Alkohol und schließlich mit Äther. Man trocknet im Vakuum bei gewöhnlicher Temperatur über P₂O₅ und unter einem Druck von etwa 20 mm Hg. Man erhält auf diese Weise 163 Teile Dinatrium-phosphokreatin-Hexahydrat, also in einer Ausbeute von 90%.For the cleavage of the alkali salts of phosphocreatinine, the reaction mass is filtered to remove the catalyst and the filter residue is then washed with 250 + 100 + 100 parts of water. 47.4 parts of sodium hydroxide, dissolved in 85 parts of water, are added to the filtrate. It is heated to 80 ° C. in 10 minutes. The solution is then cooled to about 10 ° C .; its pH is adjusted to 7.8 in the presence of phenolphthalein with 1182 parts by volume of η-hydrochloric acid, filtered and 9480 parts of alcohol are added to the solution. The disodium salt of phosphocreatine is deposited in the form of an oil. It is left to crystallize for 2 or 3 days at room temperature, filtered, washed with 80% alcohol, then with absolute alcohol and finally with ether. It is dried in vacuo at normal temperature over P ₂ O ₅ and under a pressure of about 20 mm Hg. In this way, 163 parts of disodium phosphocreatine hexahydrate are obtained, that is to say in a yield of 90%.

Dieses Phosphokreatin hat eine Titrationskurve, die mit der in der Literatur beschriebenen identisch ist.This phosphocreatine has a titration curve which is identical to that described in the literature.

Analyse für C₄H₈N₃O₅PNa₂ · 6 H₂O Molekulargewicht: 363Analysis for C ₄ H ₈ N ₃ O ₅ PNa ₂ · 6 H ₂ O Molecular weight: 363

Berechnet ... P 8,54, Na 12,67%;Calculated ... P 8.54, Na 12.67%;

gefunden ... P 8,54, Na 12,66%;found ... P 8.54, Na 12.66%;

P 8,77, Na 12,92%.P 8.77, Na 12.92%.

ίο Zur Umkristallisation wird das Natriumsalz des Phosphokreatins in 7 Teilen kalten Wassers aufgelöst; nach dem Filtrieren setzt man 21 Teile Alkohol hinzu und geht wie zuvor vor. Die Ausbeute bei der Umkristallisation beläuft sich auf 85%.ίο For recrystallization, the sodium salt of Phosphocreatine dissolved in 7 parts of cold water; after filtration, 21 parts of alcohol are added and proceeds as before. The recrystallization yield is 85%.

Analyse für C₄H₈N₃O₅PNa₂ · 6 N₂O Molekulargewicht: 363Analysis for C ₄ H ₈ N ₃ O ₅ PNa ₂ · 6 N ₂ O Molecular weight: 363

Berechnet: C 13,22, H 5,51, N 11,57, P 8,54,Calculated: C 13.22, H 5.51, N 11.57, P 8.54,

Na 12,67, H₂O 29,75%;Na 12.67, H ₂ O 29.75%;

gefunden: C 13,94, H 5,40, N 11,95, P 8,58,
Na 12,74, H₂O 28,91%.found: C 13.94, H 5.40, N 11.95, P 8.58,
Na 12.74, H ₂ O 28.91%.

Dieses Hexahydrat des Dinatriumsalzes des Phosphokreatins wurde unter einem Vakuum von 3 Mikron und bei 60° C so lange getrocknet, bis das Gewicht konstant blieb; dann ließ~man es an der freien Luft wieder bis zu einem konstanten Gewicht Wasser aufnehmen. Das Produkt enthielt dann 4 Moleküle Kristallwasser.
30 This hexahydrate of the disodium salt of phosphocreatine was dried under a vacuum of 3 microns and at 60 ° C. until the weight remained constant; then it was allowed to take up again in the open air to a constant weight of water. The product then contained 4 molecules of water of crystallization.
30th

Analyse für C₄H₈N₃O₅PNa₂ · 4 H₂O, Molekulargewicht: 327
Berechnet: C 14,68, H 4,89, N 12,84, P 9,48,Analysis for C ₄ H ₈ N ₃ O ₅ PNa ₂ · 4 H ₂ O, molecular weight: 327
Calculated: C 14.68, H 4.89, N 12.84, P 9.48,

Na 14,07%;
gefunden: C 14,66, H 5,78, ^12,73, P 9,17,Na 14.07%;
found: C 14.66, H 5.78, ^ 12.73, P 9.17,

Na 13,61, C 14,45, H 5,68, N 12,40%.Na 13.61, C 14.45, H 5.68, N 12.40%.

Bisweilen erhält man das Tetrahydrat schon von vornherein.Sometimes the tetrahydrate is obtained from the start.

Die Titrationskurve ist identisch mit den vorhergehenden. Diese Angaben stimmen vollkommen überein mit denjenigen von A. H. E η η ο r und L. H. Stocken (Biochem. J. 43, 1948, S. 190) sowie von Peanasky und Mitarbeitern (Biochemical Preparations V, 1957, S. 9), welche geringe Mengen des Dinatriumsalzes von Phosphokreatin nach einem anderen Verfahren hergestellt haben.The titration curve is identical to the previous one. These details agree completely with those of A. H. E η η ο r and L. H. Stocken (Biochem. J. 43, 1948, p. 190) and by Peanasky and co-workers (Biochemical Preparations V, 1957, p. 9), which small amounts of the disodium salt of phosphocreatine after a other processes.

Claims

1 21 2

Patent claim "obtained alkali salts of phosphocreatinine without

Separation of the same after filtering off the pallet

Process for the preparation of alkali salts of the medium catalyst by adding alkali and

Phosphocreatine splits by saponifying the ethyl ester by heating it to 80 ° C. .

of dibenzyloxyphosphoryl creatine in aqueous 5 phosphocreatine is a known substance. C. H.

alcoholic medium with diluted cold Fiske and Subahron (Journal of Biological

Sodium hydroxide and cyclization of the obtained Chemistry, 81, 1929, p. 620) have this compound

Sodium salt of the diester with equimolar men- isolated from the muscle and also the constitution

gen acid, based on the sodium to be released of this substance. Substance is essential

hydroxyd according to patent 1 260472 and hydrogenolysis ίο borrowed importance in phosphoric acid metabolism.

of dibenzyloxyphosphoryl creatinine of the formula Later, K. Z e i 1 e and G. Fawaz,

• '·' ■ "·. Hoppe-Seyler's Journal for Physiological Chemistry,

C ₆ H ₅ CH ₂ O NH CO 256, 1938, p. 194, small amounts of calcium

\ _r / '' 'j phokreatinat and finally AH Enno r and

■ / ■ ^ ■ ~ '' ⁼ \ ■ ■ '* 5 LAStocken, Biochemical Journal, 43, 1948,

QH ₅ - CH ₂ - O i 'N ^ —CH ₂ S. 190, also small amounts of sodium

^! 'O- - I produce phosphocreatinate tetra- or hexahydrate.

. ,,,;. CH ₃ All of these procedures are very tedious and of

poor yield. The method according to the invention

in an aqueous-alcoholic medium by means of water allows for the first time the production of the alkali metal salts

substance 'at ordinary, temperature in the presence of phosphocreatine with good yields in technical

a palladium catalyst and alkali by volume. It is of particular advantage that

Patent 1 260 473, characterized by this - dibenzyloxyphosphorylcreatinine due to its

net that the good crystallizability obtained by hydrogenolysis is achieved by the

ten alkali salts of phosphocreatinine introduced without the preparation of the starting compound

. Separate the same after filtering off the Palla or free any impurities that have arisen in the process

medium catalyst by the addition of alkali and leaves and surprisingly the cleavage of the

Heating at 8O ⁰ C splits. the hydrogenolysis obtained alkali salt of phosphor

. phocreatinine without their separation in the aqueous

■■ .-. · ■■ 30 alcoholic hydrogenolysis medium, despite the insolubility of the alkali salts of phosphocreatinine in

The subject of the German patent 1 260 472 is alcoholic media can carry out,
a process for the preparation of dibenzyloxy-. According to the process according to the invention, phosphoryl creatinine is obtained by saponifying the ethyl ester, a very pure phosphocreatine which is free from mineral dibenzyloxyphosphoryl creatine in aqueous alcohol phosphates and free creatine. The product medium with diluted cold sodium crystallizes with 4 molecules of crystal water; it hydroxide and cyclization of the sodium obtained is stable, does not dissolve and keeps very well salt of the diester with equimolar amounts of acid, in closed bottles without special reference to the sodium hydroxide to be released. Visual measures. .

The subject of the German patent 1 260 473 is a 40

Process for the preparation of alkali salts of the example

Phosphocreatinine by hydrogenolysis of the di- a) Production of the starting materials
benzyloxyphosphoryl creatinine using hydrogen

in an aqueous-alcoholic medium with an ordinary In one with a stirrer and a cooler

Temperature in the presence of a palladium catalyst 45 provided device leads to 175 parts of dibenzyl

and alkali. , oxyphosphoryl-S-methyl-isothiourea, 3000 parts

The present invention relates to a process of 80% alcohol, 26.5 parts of sodium carbonate and

for the production of alkali salts of phosphocreatine 54 parts of red mercury (II) oxide. One heats up

by saponification of the ethyl ester of dibenzyloxy for 2 hours with stirring at boiling temperature,

phosphoryl creatine in aqueous-alcoholic medium 50 After cooling, the mercury is filtered

with dilute cold sodium hydroxide and cyclimercaptide and removes the solvent under

Sization of the sodium salt of the diester obtained under reduced pressure. The residue is divided into 100

added with equimolar amounts of acid, based on the len absolute alcohol. This solution

Sodium hydroxide to be released according to Patent 1,260,472 is used to set 76.75 parts of sarcosine ethyl ester hydrochloride

and hydrogenolysis of Dibenzyloxyphosphorylkrea- 55 with a melting point of 121 to 122 ° C, dissolved

tinins of the formula _. , add 200 parts of alcohol. Leave overnight

. stand and then heated the reaction mixture on

C ₈ H ₅ - CH ₂ - O _v .NH CO reflux for 2 hours. After

\ ρ __ _N _ _c / cooling and filtering off the sodium chloride / μ. ~ \ .60 one removes the alcohol contained in the filtrate.

C ₆ H ₅ - CH ₂ - OI! N CH ₂ T _he residue is dissolved in 25 parts of alcohol,

O I whereupon 60 parts by volume of a 10N sodium hydroxide solution

CH ₃ added. The whole thing is left in for 1 hour

Touch, then add 250 parts of water and

filtered in an aqueous-alcoholic medium using hydrogen 65. The solution is made with 36 parts of acetic acid at ordinary temperature. acidified in the presence of; they are left to crystallize overnight

Palladium catalyst and alkali according to patent 1260473, stand, filtered, wash the precipitate with

wherein according to the invention is obtained by hydrogenolysis of 50 ° / _o ethyl alcohol, then with water and finally