DE2626792A1 - Adenosine-5'-phosphoric acid derivs. and their salts - prepd. by acylating adenosine-5'-phosphate and opt. desalting or salt exchanging - Google Patents

Abstract

Adenosine-5'-phosphate which is produced by reacting adenosine-5'-phosporic acid with organic base e.g. diethylamine is acylated with e.g. acid anhydride and opt. desalted or salt-exchanged to give adenosine-5'-phosphoric acid deriv. (I) or its salts. (where R' is H or acyl; R' is acyl; M is H, metal, or organic base residue). (I) is used as drug for circulatory organs or inter. of the above drug.

Description

KOWA COMPANY, LTD., Nagoya / Japan

Acyl derivatives of adenosine-5'-monophosphonic acid, methods for their Manufacture and containing them a pharmaceutical composition.

The invention relates to new acyl derivatives of adenosine-5'-mono phosphoric acid, dSe are effective as medicaments, as well as their intermediates, a process to their manufacture and pharmaceutical compositions containing these compounds with pharmacological effectiveness included.

In particular, the invention relates to acyl derivatives of adenosine-5'-monophosphoric acid or salts thereof, of the general formula wherein M represents a hydrogen atom or a member of the group consisting of metal atoms of Groups Ia, IIa and IIIb of the Periodic Table of the Elements and organic bases; R represents a hydrogen atom or an acyl group having 1 to 9 carbon atoms; and R 'represents an acyl group having 1 to 9 carbon atoms; as well as a process for the preparation of these compounds and a pharmaceutical composition containing a pharmaceutically effective amount of the above novel compound (s) and a pharmaceutically acceptable diluent or carrier. The salts include salts of metals such as lithium, sodium, potassium, calcium, magnesium or aluminum, tri-lower alkylamine salts, guanidine salts and 4-morpholino-N, '- dicyclohexylcarboxyamidine salts.

Adenosine-5 is present in the animal kingdom as well as in humans '-phosphates, such as adenosine-5'-triphosphate (ATP), adenosine-5'-diphosphate (ADP) or Adenosine 5'-monophosphate (Mir), which take part in the metabolism in the living body, known.

In addition, are cyclic 3 ', 5'-adenosine monophosphate (CAMP) and its acyl derivatives known as derivatives of Mir. It is also known to be pharmacological Has effectiveness as the antihypertensive activity when administered intravenously and mortality delaying efficacy in hypoxic conditions intravenous administration, although its activities are less than those of ATP, and it is also known that CAMP or its acyl derivatives when administered intravenously have anti-hypertensive activity.

However, the pharmacological properties of these tend to be known Adenosine-5'-phosphates and their derivatives in general when administered orally to reduce.

It has now been found that the new acyl derivatives of the formula 1-in can easily be prepared from Mir and that these acyl derivatives even superior pharmacological activities such as anti-hypertensive when administered orally Efficacy or an efficacy that prolongs mortality or sticking exhibit under hypoxic conditions. It has also shown that these new acyl derivatives (AI4PA) have a pharmalcological effect on the central nervous system, an effect of prolonging sticking or mortality under low pressure conditions, one pharmacological activity on cardiac failure; one on metabolism activating effect and anti-ulcerogenic activity in Reserpinulcus (mouse), Stress ulcer (rat) and Shay ulcer (rat) show what they show promise Makes use as an antialcus agent.

The present invention is therefore to create new Acyl derivatives of adenosine 5'-monophosphate.

Another object of the invention is to provide a method to make these new compounds.

Another object of the invention is to provide a pharmaceutical A composition comprising a pharmaceutically effective amount of the novel acyl derivatives of Contains adenosine 5'-monophosphate with the good pharmacological properties.

Further objects or objects and advantages of the invention are outlined apparent from the following description.

The AMPA (I) according to the invention can be prepared by reacting organic base salts of adenosine 5'-monophosphate represented by the following formula wherein M 'is a hydrogen atom or an organic base, with the proviso that two radicals M' cannot simultaneously represent a hydrogen atom, can be prepared with an acylating agent. Optionally, the product can readily be converted to a compound of Formula I wherein M is a metal from Groups Ia, IIa or IIIb of the Periodic Table of the Elements, using a salt exchange reaction. A salt elimination reaction of the above compounds leads to the conversion to the free form of their acids.

The organic base salts of Formula II can easily be prepared by reacting AMP with an organic base in an inert organic solvent. The reaction to form the salts of organic bases of the formula II from AMP can be shown schematically as follows: NHp N1i ("Y" N O organic bage 0 .. organic basve " HO-PO O inert organic 014 ' 01i cal solvent w ith solvent \ L OH HO OH (II) The reaction to form the salts of organic bases of adenosine 5'-monophosphate of the formula II from Mir is preferably carried out with heating. For example, it can be carried out at a temperature of about 30 to about 1000G. Examples of suitable organic bases are organic amines such as di-lower-alkylamines, tri-lower-alkylamines, di-lower-alkyl-lanilines, tri-lower-alkanolamines, quaternary ammonium hydroxides or substituted guanidines. Specific examples of these amines are diethylamine, triethylamine, tributylamine, trioctylamine, triethanolamine, ammonium hydroxide, tetraethylammonium hydroxide and 4-morpholino-N, N'-dicyclohexylcarboxyamidine.

Inert solvents that can be used are those that are suitable for the amines dissolve, however, with the reactants and the compound of formula II do not enter into a reaction, for example water, anhydrous pyridine, anhydrous picoline, anhydrous butidine, and dimethylformamide.

Of these, most preferred is anhydrous pyridine.

The salt of the organic base of adenosine 5'-monophosphate of the formula II can optionally be isolated in the form of crystals by evaporating the solvent and treating the residue with an organic solvent such as alcohols, acetone, ether or benzene. When carrying out the process according to the invention, however, it is not necessary to isolate the compound of the formula II, but rather the resulting reaction mixture can be subjected directly to an acylation reaction. The acylation reaction can easily be carried out by contacting the compound of Formula II with an acylating agent in an inert anhydrous organic solvent. The reaction to form the compound of formula I from the compound of formula II can be illustrated schematically as follows: NH NHR N I Ij \ N NNN OA ylation agent 0 'OPO 9 inert-organi- M'OPO 0 (cal solution ohm 4 middle HO OH RO ORe ((II),) ((I) al If it is desired to convert M 'in the compound of the formula Ia into a radical of the definition M of the formula I different from M1, a salt exchange reaction or a salt elimination reaction is carried out to convert the compound of the formula Ia into a compound of the formula Ib. NFlR NlIR NW b / NN t / Salt exchange reaction or 0 ~~~~~~~~~~~~~~~~~~~~~~~~ 1? M'o-PO o e.g. treatment with, ° \ o} Ion exchange resin OM 4 R'O OR 'R'O - ° RB ((Ia)) ((Ib)) (wherein M "is a hydrogen atom or a metal atom belonging to Groups Ia, IIa or IIIa of the Periodic Table of the Elements) Since the acylation reaction proceeds even at room temperature, it is not absolutely necessary to heat the reaction system. However, if necessary, the reaction can be carried out at an elevated temperature , such as up to ~ 100 ° C. The reaction time is not particularly limited and, for example, the reaction can be carried out from about 2 hours to about 8 days Allow the mixture to stand for about 1 to 2 days. The reaction mixture can be concentrated under reduced pressure to remove the inert organic solvent, thereby obtaining the compound of the formula Ib. Preferably, the compound is further extracted successively with ether and chloroform. The chloroform layer can be extracted under reduced pressure Pressure can be concentrated, whereby the connection derives the formula Ia. By suitable selection of the reaction conditions in the acylation reaction, compounds of the formula Ia in which R is a hydrogen atom, ie in which the amino group in the 6-position is not acylated, can be formed.

The acylating agent used in the above reaction is preferably an acid anhydride or an acid halide.

The use of acid anhydrides is preferred. It includes straight chain or branched chain aliphatic carboxylic acid anhydrides having 2 to 9 carbon atoms and straight-chain or branched-chain aliphatic carboxylic acid halides with 1 up to 9 carbon atoms. Examples of the acylating agents include formic acid-acetic acid mixture, Acetic anhydride, propionic anhydride, butyric anhydride, isobutyric anhydride, Pentanoic anhydride, pivalic anhydride, hexanoic anhydride, heptanoic anhydride, Octanoic anhydride, formyl chloride, acetyl chloride, propionyl chloride, butyryl chloride, Isobutyryl chloride, pentanoyl chloride, pivaloyl chloride, hexanoyl chloride, heptanoyl chloride and octanoyl chloride.

The compound of the formula Ia can be converted into a compound of the formula Ib converted using a salt exchange reaction or a salt elimination reaction will. This reaction can be easily carried out, for example, by using a neutralization method or an ion exchange resin method. In the neutralization method the compound of formula Ia with an aqueous solution of a metal halide, Metal hydroxide or metal carbonate, etc. are reacted to convert them into a metal salt. In addition, the compound of the formula Ia can be converted into a MirA derivative of the free acid type, in which two radicals M 'in the formula I are hydrogen atoms, taking into account the salt elimination reaction using an ion exchange resin served by the hydrogen type. Can also be used in the salt exchange reaction an ion exchange resin of the sodium type salts can be prepared, wherein M in of formula I is sodium.

The above salt exchange reaction or salt elimination reaction can by dissolving the compound of formula Ia in, for example, water or a 50% by volume aqueous solution of ethanol, passing the solution through a with a Ion exchange resin filled column and subsequent elution of the adsorbed Materials carried out with an eluting solvent using water will. Preferably the adsorption-elution operation is carried out at room temperature or below performed. The eluting operation is preferably carried out at a low one Temperature of about 5 + 500 carried out.

For the salt exchange or the salt elimination reaction, different Ion exchange resins of H-type and alkali metal or alkaline earth metal type are used will. For example, weakly acidic ion exchange resins such as Amberlite IRC-50, CG-50, IRP-64 and IRP-88 (Rohm + Haas Co.) and Duolite US-101, ES-62 and ES-63 (Diamond Alkali Co.).

Examples of the compounds of the formula I, by the inventive Methods that can be obtained are listed below.

1. O2 ', O3 "-Diformyl-adenosine-5'-phosphate 2. N6, O2', O3'-triformyl-adenosine-5'-phosphate 3. O2 ', O3'-diacetyl-adenosine-5'-phosphate 4. N6.02, 0 -triacetyl-adenosine-5'-phosphate 5. O2 ', O3'-dipropionyl-adenosine-5'-phosphate 6. N6, O2', O3'-tripropionyl-adenosine-5'-phosphate 7. O2 ', O3'-dibutyryl-adenosine-5'-phosphate 8. N6, O2', O3'-tributyryl-adenosine-5'-phosphate 9. O2 ', O3'-diisobutyryl-adenosine-5'-phosphate 10. N6, O2', O3'-triisobutyryl-adenosine-5'-phosphate 11. O2 ', O3'-dipivaloyl-adenosine-5'-phosphate 12. N6, O2', O3'-tripivaloyl-adenosine-5'-phosphate 13. O2 ', O3'-dipentanyol-adenosine-5'-phosphate 14. N6, O2', O3'-tripentanoyl-adenosine-5'-phosphate 15. O2 ', O3'-dihexanoyl-adenosine-5'-phosphate 16. N6, O2 ', O3'-Trihexanoyl-adenosine-5'-phosphate 17. O2 ', O3'-Diheptanoyl-adenosine-5'-phosphate 18. N6, O2', O3'-Triheptanoyl-adenosine-5'-phosphate 19. O2 ', O3'-dioctanoyl-adenosine-5'-phosphate 20. N6, O2', O3'-trioctanoyl-adenosine-5'-phosphate 21. O2 ', O3'-Dinonanoyl-adenosine-5'-phosphate 22. N6, O2', O3'-trinonanoyl-adenosine-5'-phosphate 23. Sodium, calcium, magnesium, aluminum, guanidine and triethylamine salts of compounds 1 to 22.

The pharmacological properties of some of the new invention Connections are shown below.

The numbers of the compounds refer to the tables below to the above numbers or connections.

The evaluation was carried out using a scale a, b and c, wherein a means that the pharmacological activity is superior, b means that the pharmacological activity is recognizable, but under a and c means that the pharmacological effectiveness is negligible.

Table I Antihypertensive Activity attachment rating Sodium salt of 7. b Sodium salt of 8. a Sodium salt of 10. a Guanidinium salt of 10 a ATP (sodium salt) # b AMP (sodium salt) # b Test animal: rat Test method: The compound was administered orally, and the blood pressure on the carotid was measured by an electric hemodynamometer.

Table 2 Antiulcerogenic Efficacy Compound Reserpinulcus of the mouse Shay.Ulcus d.Ratte (oral administration, (oral administration 5 mg / kg) Sodium salt of 8th Sodium salt of 10 ATP cc AMP cc Control attempt (physiological saline solution) cc Table 3 Mortality under hypoxic conditions Compounds Intravendse Injection Oral Administration Sodium salt v.6. away Sodium salt of 8. aa Guanidinium salt x8. aa Sodium salt v.9. away Sodium salt of 10. aa Guanidinium salt vD aa ATP ac aab ontroll attempt (Salcoseryl) - c The experiments were carried out on the mouse. A gaseous mixture of 94 N2 and 6% 02 was introduced and the time until the code entered was measured.

Table 4 Tranquilizers (Minor) Compound taming effect potentiating the (Mouse, po) thiopental sleep (Mouse, po) Sodium salt of 8th Sodium salt of 10. aa Guanidinium salt of 10 aa AMP cc ATP cc The acute toxicity of the new compounds according to the invention is very low and is around 4000 mg / kg when administered orally to the rat.

The new AWirA compounds according to the invention and their salts can be set in the form of a pharmaceutical composition, the one pharmaceutically effective amount of such a compound and a pharmaceutically usable resp.

contains a compatible diluent or a carrier.

Examples of such carriers and diluents are solid diluents or carriers such as starch, lactose, sucrose or sucrose, glucose, mannitol, dextrin, Gum arabic, sodium alginate, gelatin, methyl cellulose, ethyl cellulose, hydroxyethyl starch, Polyvinyl pyrrolidone and polyvinyl alcohol.

An example of a liquid diluent or liquid The carrier is purified water for injection.

The amount of the compound of the invention in the pharmaceutical Composition can be any and is generally from about 0.01 to about 99.1% by weight. The dosage of the compound according to the invention varies depending on the disease to be treated and is, for example, about 0.5 to about 100 mg / kg body weight / day

The following examples serve to illustrate the invention.

Example 1 Sodium O² ', O³'-Dibutylryl-adenosine-5'-phosphate 347 mg (1 mole) of adenosine-5'-phosphoric acid and 293 mg (1 mole) of 4-morpholine-N, N'-dichlorohexylcarboxyamidine were suspended in 10 ml of anhydrous pyridine and the suspension was over a boiling water bath to form a 4-morpholino -N, N'-dichlorohexylcarboxyamidine salt of adenosine-5'-phosphoric acid heated. The reaction mixture was cooled to room temperature and 5 ml of butyric anhydride was added and the mixture was stirred for about 7 hours, whereby the reaction mixture became clear. The reaction mixture was then cooled with ice and, after 10 ml of water had been added, left to stand overnight at room temperature. The mixture was then concentrated under reduced pressure and 80 ml of ether was added. The mixture was extracted five times with 20 ml of water. The resulting aqueous layer was extracted six times with 20 ml of chloroform. The chloroform extract was concentrated under reduced pressure and chloroform was added. The mixture was azeotroped to remove water. The residue was dissolved in a small amount of chloroform, and 100 ml of ether was added dropwise to the solution. By filtering the mixture, 350 ml (yield 45) of 4-morpholino-N, N'-dicyclohexylcarboxyamidine salt of O² ', O³ -dibutyryl-adenosine-5'-phosphate was obtained in the form of a white powder.

100 mg of this powder was dissolved in 50% aqueous ethanol and the mixture was passed through a column of Amberlite IRC-50 (ion exchange resin from Na type from Rohm + Haas Co.). The adsorbed material was washed with water eluted.

50 ml of the first fraction was collected and placed under reduced pressure concentrated. A small amount of ethanol was added to the concentrated solution and the mixture was added dropwise to 200 ml of ether, giving 62 mg (yield 85%) sodium 02,03,0³'-dibutyryl-adenosine-5'-phosphate in the form of a white powder received.

The following analysis results of the product were obtained: Elemental analysis for C18H24N5O9PNa2 # 2H2O C H N P Calculated (%): 38.10 4.97 12.34 5.46 Found (O / o) : 38.24 4.69 12.10 5.57 Paper Chromatography (PPC): Rf value 0.58 Toyo filter paper No. 51, descending method.

Developing solvent: ethanol / 0.5 m ammonium acetate = 5: 2 Determination method: 1. Benrei quartz lamp (short-wave UV) (+) 2. Phosphor color (+) 3. Benzidine color (-) Thin layer chromatography (TLC: Rf value 0.36, PEI-Celulose F (solvent and determination method were the same as for paper chromatography).

UV absorption spectrum # maxH2O: 260 mµ (# 1.24 x 104) + #maxn: 257.5 mµ # maxOH-: 260 mµ Example 2 Sodium N6, O² ', O³'-tributyryl-adenosine-5'-phosphate : 347 mg of adenosine-5'-phosphoric acid were dissolved in 2 ml of a 0.4 aqueous solution of triethylamine and the solution was evaporated to dryness under reduced pressure. Anhydrous pyridine was added to the residue and the mixture was evaporated to dryness three times under reduced pressure. The product was further dried under reduced pressure for 5 hours in a desiccator using phosphorus pentoxide.

10 ml of anhydrous pyridine were added to the dried product and 5 ml of butyric anhydride. The solution was left to stand at room temperature for 8 days and then extracted three times with 30 ml of ether and six times with 30 ml of chloroform. The resulting chloroform layer was concentrated under reduced pressure and after the addition, azeotropically distilled until no more odor of pyridine is tested was. The residue was dissolved in a 50% aqueous solution and the solution was passed through a column of Amberlnte IRC-50 (Na type ion exchange resin). The adsorbed material was eluted with water. 200 ml of the first discharge were collected and concentrated under reduced pressure. 200 ml of acetone were added added to the concentrated liquid and the mixture was left overnight at room temperature ditched. The precipitate was separated by centrifugation, whereby 448 mg (yield 70%) sodium N6, O² ', O³'-tributyryl-adenosine-5'-phosphate in Porm white crystals of m.p. = 171 to 1740C.

The product was elysed. The following results were obtained: Elemental analysis for C22H30N5010PNa2 2H20 C H N P Calculated (%): 41.45 5.38 10.99 4.86 Found (): 41.68 5.18 10.79 4.80 Paper chromatography: Rf value 0.78 The developing solvent and the determination method was the same as in Example 1.

Thin layer chromatography: Rf value 0.58 UV absorption spectrum: # H2Omax: 274 mμ (# 1.69 x 104) H + # max @ 213 mμ # maxOH-: 274 mμ Examples 3 to 11 Examples 1 and 2 were never used under the conditions given in Table 1. The properties of the compounds obtained are also shown in Table 1. In the table, G has the meaning of 4-morpholino-N, N'-dichlorohexylcarboxyamidine and TEA means triethylamine. Table 1 Uv Spec- <1> dream Rf XR, 110 2 GI Am2r Connection PPC TLC (all) 3 Sodium1T6,021,03t -triacetyl- o.48 os33 273-; 5 adenosine-5'-phoht Sodium-N, 0, Q 031-tripropionyl 4 adenosine 5'-phosphate o164 0.40 273.5 Sodium 02,031-diisobutyryl- 5 ademosin-5 phosphate O56 0.38 260 Sodium-lJ6sQ2, Q3 -tri isobutyr-. 6 yl-adenosine-5-phosphate 0.80 0.58 274 Sodium N6,02,03 -tripivaloyl- 7 adeno.it-5'-phosphate - 0.57 274. Sodium-O, 0 '-dnhexanoyl- -adonosine 5'-phosphate 0.56 0.38 259.5 atrium-N6,021,031-trihexanoyl-ear58 274 adenosn-5'-Fkosphet 01 Fatrit7m-02, 03'-dioctanoyl- 10 adenosine-5-phosphate 0.67 o.4o a59.5 Itodium-N6,021,03'-triootanoyl ll adenosnn-5t-phosphate 0.78 0.58 274 Table 1 continued H reaction conditions Q> PI 5'-AMP Use ReactiOflReaktionr R (Nol) - te base tempera - time be; zt e. ture (P) Room- 2 days 68 182-184 temp. Zrs,). 4 15 TEA 1I 3 n 42 179-181 ( 1 ) 5 2 G <1 5 hours 46 6 3 G .. 8 days 78 156-159 (Decomp.) 7 - 3 TEA 1000C 3 hours 15 206-209 (1l) 8 2 G 'Raun- 5 "41 temp. 9 3 TEA .. 8 days -6o 141-143 10 3 b It 2 hrs. 36 11 3 G fl. 7 days 52 Example 12 3.185 g (5 mmol) of disodium N6, O² ', O³'-tributyryl-adenosine-5' phosphate were dissolved in 150 ml of cold water and 110 ml of an aqueous solution of AlOl3.6H20 (1.61 g / 200 ml H20 ) was slowly added dropwise with vigorous stirring. After the addition, the precipitate was filtered off, washed with water and dried under reduced pressure over phosphorus pentoxide in a desiccator. The dried product was then dissolved in acetone and filtered. The filtrate was added dropwise to 200 ml of water. The resulting precipitate was collected by filtration and dried over phosphorus pentoxide in a desiccator to give 2.6807 g (94.3% yield) of aluminum-N6, melting point = 229-231 ° C (charred).

The product was analyzed and the following results were obtained: UV absorption spectrum: #maxEtOH: 273.5 (# 14.7 x 10³) thin layer chromatography (TLC): Rf value = /, 94 PEI cellulose F (ethanol: 1 m-ammonium acetate = 5: 2

Claims (6)

Claims (1.) Acyl derivatives of adenosine-5'-monophosphonic acid or their se of the following formula: wherein M is a hydrogen atom or a member of the group consisting of metal atoms of Groups Ia, IIa and IIIb of the Periodic Table of the Elements and of organic bases; R represents a hydrogen atom or an acyl group having 1 to 9 carbon atoms; and R 'is an acyl group having 1 to 9 carbon atoms. 2. Compounds according to claim 1, wherein M is selected in the formula is from the group of lithium, natriurl, potassium, calcium, magnesium, aluminum, tri-lower alkylamines, guanidines and 4-morpholino-N, N'-dicyclohexylcarboxyamidine. 3. Compounds according to claim 1, wherein the acyl group represented by R is selected from the group of formyl, acetyl, propionyl, butyryl, pentanoyl, Hexanoyl, heptanoyl and octanoyl. 4. Compounds according to claim 1, wherein that represented by R ' Acyl group is selected from the group of formyl, acetyl, propionyl, butyryl, Pentanoyl, hexanoyl, heptanoyl and octanoyl. 5. A pharmaceutical composition containing 1. an acyl derivative of adenosine-5'-monophosphoric acid or its salt according to one of the preceding claims with the formula wherein M is a hydrogen atom or is selected from the group of atoms of groups Ia, IIa and IIIb of the Periodic Table of the Elements and organic bases; R is a hydrogen atom or an acyl group having 1 to 9 carbon atoms; and R 'is an acyl group having 1 to 9 carbon atoms, and 2. a pharmaceutically usable or compatible diluent or a pharmaceutically usable or compatible carrier, or customary auxiliaries and carriers. 6. A process for the preparation of an acyl derivative of adenosine-5'-monophosphonic acid or its salt according to any one of claims 1 to 4, having the following formula: wherein M is a hydrogen atom or is selected from the group of metal atoms of groups Ia, IIa and IIIb of the Periodic Table of the Elements and organic bases; R is a hydrogen atom or an acyl group with 7 to 9 carbon atoms and Rt e & ne is acyl group with 1 to 9 carbon atoms, characterized in that an organic base salt of adenosine-5'-monophosphonic acid of the following formula: in which M 'is a hydrogen atom or an organic base, with the proviso that the two symbols M' are not hydrogen atoms at the same time, reacts with an acylating agent and optionally subjects the product obtained to a salt exchange reaction or a salt elimination reaction.