GB2044265A

GB2044265A - Adenosine Triphosphate Metal Complexes, Processes for Their Preparation and Pharmaceutical Compositions Containing Them

Info

Publication number: GB2044265A
Application number: GB7908428A
Authority: GB
Original assignee: Chinoin Gyogyszer es Vegyeszeti Termekek Gyara Zrt
Current assignee: Chinoin Private Co Ltd
Priority date: 1979-03-09
Filing date: 1979-03-09
Publication date: 1980-10-15
Also published as: GB2044265B

Abstract

Adenosine triphosphate metal complexes have been found to influence the exchange of energy and metabolism of cells. The physiologically compatible complexes are of interest in the treatment of cerebrovascular and cerebrosclerotic diseases and are of assistance in the recovery of bone tissue in the case of fractures as well as in osteoporesis and cysts in bones. The adenosine triphosphate metal complexes may be prepared by reacting adenosine triphosphate or a salt thereof with the metal or a salt thereof preferably in aqueous solution. The preparation of the complexes of the invention and pharmaceutical compositions containing the complexes as active ingredient are exemplified.

Description

SPECIFICATION Adenosine Triphosphate Metal Complexes, Processes for Their Preparation and Pharmaceutical Compositions Containing Them The present invention relates to adenosine triphosphate metal complexes, processes for their preparation and pharmaceutical compositions containing them. The complexes of the present invention possess interesting physiological activity.

Thus according to one feature of the present invention there are provided adenosine triphosphate metal complexes.

The term complex as used herein means a complex formed with a positive metal ion. All metals of the periodic system may be used as the metal ion.

Particularly preferred metals include the alkaline earth metals, for example, calcium, strontium and barium; transition metals, such as metals of Group Vlla of the Mendeleev periodic table, metals of Group VIII of the Mendeleev periodic table e.g. the iron group of elements such as iron, cobalt and nickel; rare earth elements; metals of Group Ib of the Mendeleev periodic table of elements; metals of the platinum group e.g. platinum, ruthenium, palladium or iridium, and metals belonging to the actinide group (uranium group) of elements as well as metals in Group llb of the Mendeleev periodic table of elements. Metals of the iron group e.g. iron, cobalt or nickel are regarded as most important in view of their biological activity.

Metals such as iron, cobalt and nickel belong to the iron group and metals such as platinum, ruthenium, palladium and iridium belong to the platinum group. Preferred actinide elements are uranium and thorium. Among the elements belonging to Group Ib of the periodic system copper is especially preferred, among elements belonging to group llb zinc is most preferred and among elements belonging to group Vlla manganese is most preferred.

Particularly preferred complexes according to the present invention include: Adenosine triphosphate-manganese complex, Adenosine triphosphate-cobalt complex, Adenosine triphosphate-nickel complex, Adenosine triphosphate-copper complex, Adenosine triphosphate-zinc complex, Adenosine triphosphate-uranyl (UO2) complex, Adenosine triphosphate-chromium complex, Adenosine triphosphate-iron complex, Adenosine triphosphate-thorium complex, Adenosine triphosphate-calcium complex, Adenosine triphosphate-strontium complex.

The complexes of the present invention are preferably substantially free of co-occurring substances found in nature, if any and may especially be in substantially pure form.

It will be appreciated that the complexes for use in pharmaceutical compositions are the physiologically compatible complexes. Other complexes may, however, be used in the preparation of the physiologically compatible complexes.

The preparation of adenosine diphosphate chrome (Ill) complexes is disclosed in Probes Struct.

Funct. Macromol. Membranes, Proc. Colloq. Johnson Res. Found. 5th, 1969 (publ. 1971), 1, p. 109- 123. The complex is prepared by oxidation of the corresponding chrome (II) complex. The preparation of the complexes of adenine and adenosine is disclosed in Biochemistry 1971, 10(20), 2669-3679.

According to the above references the complexes were only prepared for physico-chemical testing, there being no disclosure of any pharmacological activity.

In the cells of the higher orders of living organisms energy is supplied by respiration. The energy in the cells is generally transmitted by adenosine triphosphate. The adenosine triphosphate (ATP) transmits the energy released during respiration or digestion to different parts of the cell. Adenosine triphosphate plays a key-role in many processes, for example in metabolic processes such as conversion of nutrients, in mechanical processes such as muscle movement and in the resorption of substances in the intestines and in the kidney.

It is believed that ATP takes part in more than 210 primary biochemical functions, such as the transfer of groups, oxidative phosphorylation. DNA and RNA synthesis, protein synthesis, muscle contraction, stimulus formation and stimulus transfer, transport through cell membranes, control of the pH of cells and of osmotic pressure, calcium ion transfer (i.e. information transmission between cells), chromosome division etc.

Macroelements such as Ca2+ and Mg2+, and Na+ and K+ are found in the living ceil. All the other metal ions which may be found in the cells are called trace elements and the presence of many of these elements is essential to the living organism. These elements are important mainly in coenzymes. The ATP molecule contains its usuable energy accumulated in the bonds between phosphorus and oxygen atoms (7-1 5 Kcal/mole), these bonds being kinetically stable. We have found in our experiments that the energy state of the ATP molecule may be induced into two extreme energy states (activated or deactivated) by the use of certain types of radiation, for example x-radiation and ultrasonic radiation.

Due to the activity of its specific enzyme the activated ATP molecule thus obtained releases the energy accumulated in the PO bond very rapidly, whereas the energy emission of the deactivated molecule is very slow, or ceases in extreme cases.

The activated and deactivated ATP molecules could not be prepared effectively by radiation, however, because of the low yield.

We have found that the polarity of the phosphorus and oxygen atoms may be changed as a result of certain photosensitization effects and due to hydrogen bridge bonds formed between the different parts of the ATP molecules. It is thus obvious that the electron distribution between the phosphorus and oxygen atoms and other electron effects have changed. The present invention is based on the discovery that these effects (i.e. changing the bond relationships of phosphate esters of high energy level) can be achieved by the formation of complexes with paramagnetic metal ions. Thus it is possible to prepare an ATP molecule population in which the members differ in energy distribution and thus in fine structure.The appropriate ATP-homologues are capable of influencing the exchange of energy and metabolism of the cell by acting upon the corresponding receptors of the variously differentiated cells.

By forming complexes the trace elements having a specific biological activity may be transmitted to the appropriate receptors of the cells in the form of an ATP complex. The metal ion will be localized on the receptors, and its biological activity optionally in radioactive form may be tested. Another advantage of complex formation is that the trace element may be stabilized at a lower (biologically more valuable) valency state, and thus it may be protected from different physico-chemical attacks during transport from the place of entry into the organism to the receptors at the destination cell.

According to a further feature of the present invention there is provided a process for the preparation of adenosine triphosphate metal complexes as hereinbefore defined which comprises reacting adenosine triphosphate or a salt thereof with a metal or salt thereof whereby an adenosine triphosphate metal complex as hereinbefore defined is obtained.

The reaction is preferably effected using a metal salt, advantageously a water-soluble metal salt.

Where the adenosine triphosphate is used in the form of a salt, the salt is preferably a water-soluble salt. The reaction is preferably effected in aqueous solution, advantageously at a temperature of from 0 to 1 OOC. The adenosine triphosphate metal complex may, for example, be precipitated from the reaction mixture e.g. with acetone. The reaction yield may be about 65 to 85% and the product is in a well defined solid crystalline form.

The compounds of the invention exhibit a wide spectrum of activity. In particular the complexes of the present invention influence the energy level of the living cells i.e. the metabolism of the organism. Thus the complexes are of interest in the treatment of cerebrovascular and cerebrosclerotic diseases, the recovery of bone tissue in cases of bone fractures, the treatment of osteoporesis, and of cysts in bones, and the stimulation, inhibition or complete elimination of the function of certain differentiated cells.

Thus, for example, the ATP-Co complex of the present invention may, for example, be administered intravenously or intramuscularly, conveniently at a daily dosage of 5-10 mg (the cerebrovascular diseases) to treat patients suffering from ischaemic and anoxic damage. Clinical examination showed that when administered intravenously or intramuscularly 67.6% of the treated patients became suitable for rehabilitation, administration of Complamin (xanthinol niacinate) resulted in rehabilitation in 72.4% of the cases, and Xavin resulted in rehabilitation in 47.5% of the cases. In this connection it should be noted that the complex of the invention was only used for the treatment of patients, in which administration of Xavin or Complamin proved ineffective.Moreover it was possible to revive three out of four persons who hanged themselves by intrathecal treatment with the ATC-Co complex of the present invention after the onset of clinical death.

The complexes of the present invention show a double activity in that they possess both a vasodilating activity and an ability to increase the metabolic state i.e. the energy state of the cerebral cells, whereas the known compounds Xavin and Complamin only exhibit a vasodilatating activity.

According to a further feature of the present invention there are provided pharmaceutical compositions comprising as active ingredient an effective amount of at least one ATP-metal complex as hereinbefore defined in association with a pharmaceutical carrier or excipient.

The complexes of the present invention may be formulated into pharmaceutical compositions by conventional methods. The pharmaceutical compositions may be presented in forms suitable for oral, subcutaneous or intravenous administration and may, for example, contain pharmaceutically compatible lubricants, flavouring agents or other excipients. The complexes of the present invention are preferably presented in the form of pharmaceutical compositions comprising at least one ATPmetal complex as hereinbefore defined in association with a sterile pharmaceutical carrier or excipient e.g. in the form of sterile solutions. In this connection the complexes of the present invention are preferably administered intravenously, e.g. dissolved in a physiological saline solution. Since most of the complexes of the invention are water soluble, injectable solutions may readily be prepared, but the complexes may also be administered in solid form, for example tablets, pills and capsules etc.

The compositions of the present invention are preferably presented in dosage unit form (as hereinafter defined) e.g. in the form of tablets, pills, capsules, vials or ampoules. Each dosage unit preferably contains from 1 to 1 50 mg, especially from 1 to 1 00 mg of active ingredient.

The term "dosage unit" as used herein means a pharmaceutical composition of the present invention in a form adapted to provide a single or unitary dosage of the said active ingredient. Thus, for example, tablets, pills, capsules, vials and ampoules may be in a form adapted to provide a single or unitary dose of the active ingredient and thus such tablets, pills, capsules, vials and ampoules may be considered to be in dosage unit form.

A preferred pharmaceutical composition according to the present invention comprises the adenosine triphosphate cobalt complex as active ingredient.

The following Examples illustrate the present invention: Example 1 10 g of ATPNa2 and an equivalent amount of a metal salt (MnSO4.4 H20:3.67 g., CoCI2. 6 H2O:3.93 g., NiCI2. 6 H2O:3.93 g., CuCI2. 2 H2O:2.80 g., ZnCl2:2.25 9.) are separateiy dissolved in 50 mi of water at about 4 to 5 C, filtered if necessary, and then mixed together. The mixture is poured into 100 ml anhydrous acetone cooled by a mixture of salt and ice, whereupon a compact precipitate is obtained. The precipitate is rapidly filtered through a glass filter which can be cooled by applying strong suction. The substance thus obtained is dried in a vacuum dessiccator over P205.It can be filtered by suction several times a day and the substance slowly crystallizes. The dry product is pulverized.

Yields of the metal complexes: Mn: 72% Co: 82% Ni: 80% Cu: 78% Zn: 65% Complexes of ATP with UO2+, Cr, Fe, Th, Sr and Ca ions may similarly be prepared by using an equivalent amount of the corresponding metal salt which is preferably a water soluble metal salt.

C% HO/o NO/o calc. found calc. found calc. found Mn2±ATP 21.44 20.84 2.52 2.62 12.5 11.98 Co2±ATP 21.29 20.98 2.50 2.58 12.42 11.90 Ni2±ATP 21.29 20.74 2.50 2.60 12.42 12.00 Cu2±ATP 21.12 20.82 2.48 2.57 12.33 11.96 Zn2±ATP 21.05 20.80 2.47 2.58 12.27 11.85 UV spectrum: absorption maxima of 0.1 moie of aqueous solution Mn2±ATP 258 nm log =4.15 Co2±ATP 258 nm log =4.25 Ni2±ATP 258 nm log E =4.25 Cu2±ATP 258 nm log E =4.22 Zn2±ATP 260 nm log E =4.16 iR spectra: in the range of 375-875 cm-l (KBr technique, SP-100-Unicam spectrophotometer): ATP Mn2±A TP C02±ATP Ni"±A TP Cu2±ATP Zn2±ATP - 555 550 535 550 545 - 630 640 625 630 640 708 - - - - 727 729 725 728 730 728 826 831 831 834 834 833 P-O-P 919 932 920 924 926 929 PO32- 980 - - P-O+C-O 1006 1009 1014 1012 1016 1020 1050 1112* 1132* 1110* 1097* 1115* 1110* P.O 1265 1240 1245 1247 1234 1250 1421 1432 1432 1440 1439 1434 1504 1520 1518 1528 1524 1520 C-N 1618 1632 1626 1628 1631 1629 NH 1660 1662 1662 1664 1664 1665 1718 1709 1708 1711 1710 1709 NH 3370 3380 3370 3380 3390 3380 UO2±A TP Cr2±ATP Fe2±A TP Th2±A TP Sr2±ATP Ca2±ATP 540 515 532 530 524 525 645 625 623 642 640 640 725 724 725 723 722 730 830 830 830 830 825 825 P-O-P 920 - 930 920 940 940 P-O+C-O 1000 1060 1000 1000 1085 1085 1100 1130 1114 1120 1120 P-O 1260 - 1235 1235 1230 1438 1430 - 1430 1438 1435 1518 - 1516 1520 1520 1552 C-N 1617 - 1620 1620 1620 1620 NH 1652 1660 - 1650 - - 1704 1712 1697 1700 1700 1700 NH 3200- 3200- 3200-- 3100-- 3200- 3200- 3600 3500 3500 3500 3500 3500 Magnetic moment: Electromagnet of type Weiss, Gouy method, 250C. measured in solid state: (8N magnetic Metal complex spin moment moment) Mn2±ATP 5.92 5.89 Co2±ATP 3.88 4.85 Ni2±ATP 2.83 3.12 Cu2±ATP 1.73 2.08 Zn2±ATP O 0 (diamagnetic) The measured values correspond to the calculated values.

Dielectric constant (): Measurements in solid state: Mn2±ATP 2.517 Co2±ATP 2.701 Ni2±ATP 2.702 Cu2±ATP 2.678 Zn2±ATP 2.602 Example 2 Composition of pharmaceutical compositions containing the active ingredient of the invention: Injectable solution: ATP-Co complex 90 mg.

distilled water to 3 ml.

Tablets: ATP-U02 complex 50 mg.

lactose 190 mg.

corn starch 25 mg.

saccha rose 5 mg.

talcum 10 mg.

sodium stearate 4 mg.

Pharmacological Test Results I. Circulation-pharmacological Screening of Compounds of the Present Invention and Analysis of the Activity Thereof 1. Antianginal Activity Acute Corononary Insufficiency Induced by Glandnitrine The compounds were screened in narcotized rats on the basis of the protective activity observed against the rise of the T-wave induced by glandnitrine. A 0.1 mg/kg. dose of the different salts of ATP was administered intravenously and the antianginal activity thereof was compared with the same activity using Na-ATP.

When comparing the different salts of ATP with Na-ATP, we found that the salts showed approximately the same relative activity and a significant difference could be seen only in the case of Ni-ATP, the activity being about one tenth of the activity of Na-ATP.

2. Haemodynamical Effects Arterial blood pressure was decreased in dogs by the use of all the compounds of the present invention which were tested, and this was the case in respect of each of the three applied doses. The activity was found to be approximately the same as the activity of Papaverine.

Respiration rate was significantly increased by the tested compounds of the present invention.

The tested compounds of the present invention had no significant heart rate increasing activity.

Perfusion of a. femoralis was increased by Na-ATP, and by Ni-ATP to an extent similar to Papaverine, whereas Zn-ATP, Cu-ATP and Mn-ATP decreased perfusion approximately to the same extent.

Femoralis resistance was increased by Zn-ATP and Mn-ATP, whereas the use of Cu-ATP resulted in no change.

The carotid perfusion was increased by the tested compounds of the invention to only a negligible extent, but Ni-ATP caused an increase in perfusion similar to Papaverine.

Carotid resistance was decreased most by Ni-ATP. Ni-ATP causes the greatest increase in carotid perfusion, its activity corresponding to that of Papaverine but its time of action being almost twice as long as that of Papaverine.

Vertebral perfusion could be increased to an extent similar to that of Papaverine only by the use Mn-ATP, and duration of the action being several times longer than that of Na-ATP of Papaverine.

Vertebral resistance related to the activity of Papaverine was considerably decreased only by Co ATP.

The contractility of the heart could be increased only by the use of, Co-ATP, and Ni-ATP related to the activity of Na-ATP, this increase being greater than the increase obtained using Papaverine, but the duration of action of Papaverine was found to be three times longer.

The compounds of the present invention did not change cardiac output significantly.

Total peripheral resistance was decreased by all the tested ATP complexes of the present invention, thus, for example the activity of Ni-ATP was equal to that of Papaverine considering both intensity and duration of activity, the intensity being almost twice as great as that obtained with Na ATP.

The activity of the left heart ventricle was considerably decreased by the tested ATP complexes of the present invention.

Coronary perfusion was significantly increased by the tested ATP metal complexes of the present invention when compared with Na-ATP, the action time however was not changed.

The only exception is Zn-ATP, where the duration of activity is four times longer.

Coronary resistance was decreased by all the tested ATP complexes of the invention.

Oxygen consumption of the left heart ventricle was significantly decreased by the tested ATP complexes of the present invention.

Myocardial oxygenation has been improved by the tested ATP complexes of the present invention compared with the control Na-ATP, which improvement was accompanied by a significant increase in the available oxygen-oxygen consumption ratio.

The efficiency of the /eft heart ventricle has improved compared with Na-ATP, when Co-ATP, Zn ATP, and Ni-ATP (Table 1) were administered.

Table 1 Activity Upon Efficiency of Left Heart Ventricle of Narcotized Dog Efficiency (mkg/ml/ 100 g) Duration of Deviation from activity the basic (mien) Test-compound Dose n Basic value Changed value value ( / ) max.x totalxx Na-ATP 0.2 mg/kg 5 0.20+0.004 0.23+0.05 +15 1 6 Papaverine 0.5 mg/kg 8 0.32+0.004 0.25+0.03 -22 1 12 1.0 mg/kg 7 0.27+0.04 0.19+0.03 -30 1 11 Co-ATP 2.0 mg/kg 8 0.27+0.03 0.15+0.02 -44 1 10 Co-ATP 0.2 mg/kg 5 0.23+0.05 0.29+0.04 +26 1 4 Zn-ATP 0.2 mg/kg 5 0.19it.03 0.25+0.05 +32 1 3 Cu-ATP 0.2 mg/kg 4 0.1 9i0.03 0.20+0.03 + 5 1 8 Mn-ATP 0.2 mg/kg 4 0.19+0.03 0.21+0.04 +11 1 6 Ni-ATP 0.2 mg/kg 4 0.26+0.05 0.32+0.05 +23 0.5 3 3. Action on Isolated Heart Mitochondria The tested ATP complexes do not influence the maximum oxygen consumption rate of mitochondria at a concentration of 125 mg/ml which can be measured in the presence of CCCP.

None of the tested ATP complexes show a mitochondrial electron transport inhibiting activity at the test concentration.

The tested ATP-complexes did not significantly increase the rate of oxygen consumption, which indicates that the complexes do not possess a disconnecting activity at the concentration used.

The ATP did not significantly decrease the rate of oxidative phosphorylation, i.e. they do not show oxidative phosphorylation inhibiting activity.

At the test concentration the ATP compounds did not decrease the efficiency of oxidative phosphorylation.

The extent of potassium acetate accumulation 1, 2 and 4 minutes after the administration of 125 mg/ml of the ATP compound was measured. The activity of the tested ATP complexes was compared with the activity of Na-ATP (Reanal). The activity of the control Na-ATP was considered as 1 00%.

According to the measured values the activity of Cu-ATP is the same as that of Na-ATP, whereas when Co-ATP, Zn-ATP, Mn-ATP and Ni-ATP are used the rate of potassium accumulation in the mitochondria was only 50%. It must be noted that as in the case of potassium acetate accumulation all the tested ATP complexes also ensured potassium phosphate accumulation in the mitochondria depending upon energy.

Claims

Claims 1. Adenosine triphosphate metal complexes. 2. Complexes as claimed in claim 1 wherein the metal is in alkaline earth metal. 3. Complexes as claimed in claim 2 wherein the metal is calcium, strontium or barium. 4. Complexes as claimed in claim 1 wherein the metal is a transition metal. 5. Complexes as claimed in claim 4 wherein the metal is in Group Vlla of the Mendeleev periodic table of elements. 6. Complexes as claimed in claim 4 wherein the metal is in Group Vlil of the Mendeleev periodic table of elements. 7. Complexes as claimed in claim 6 wherein the metal is of the iron group of elements (as herein defined). 8. Complexes as claimed in claim 6 wherein the metal is of the platinum group of elements (as herein defined). 9. Complexes as claimed in claim 8 wherein the metal is platinum, ruthenium, palladium or iridium. 10. Complexes as claimed in claim 4 wherein the metal is in Group Ib of the Mendeleev periodic table of elements. 11. Complexes as claimed in claim 4 wherein the metal is of the actinide group of elements. 12. Complexes as claimed in claim 1 wherein the metal is in Group llb of the Mendeleev periodic table of elements. 13. Adenosine triphosphate-magnanese complex. 14. Adenosine triphosphate-cobalt complex. 1 5. Adenosine triphosphate-nickel complex. 1 6. Adenosine triphosphate-copper complex. 1 7. Adenosine triphosphate-zinc complex. 1 8. Adenosine triphosphate-uranyl (U 2) complex.

1. Adenosine triphosphate metal complexes other than such complexes wherein the metal is sodium or magnesium.

1 9. Adenosine triphosphate-chromium complex.

20. Adenosine triphosphate-iron complex.

21. Adenosine triphosphate-thorium complex.

22. Adenosine triphosphate-calcium complex.

23. Adenosine triphosphate-strontium complex.

24. Complexes as claimed in claim 1 wherein the metal is paramagnetic.

25. Complexes as claimed in any one of the preceding claims substantially free of co-occurring substances found in nature, if any.

26. Complexes as claimed in any one of the preceding claims in substantially pure form.

27. A process for the preparation of adenosine triphosphate metal complexes which comprises reacting adenosine triphosphate or a salt thereof with a metal or salt thereof whereby an adenosine triphosphate metal complex is obtained.

28. A process as claimed in claim 27 wherein the reaction is effected using a metal salt.

29. A process as claimed in Claim 28 wherein the metal salt is a water soluble metal salt.

30. A process as claimed in any one of claims 27 to 29 wherein the reaction is effected using a water soluble salt of adenosine triphosphate.

31. A process as claimed in any one of claims 29 to 30 wherein the salt is a chloride or sulphate.

32. A process as claimed in any one of claims 27 to 31 wherein the reaction is effected in aqueous solution.

33. A process as claimed in any one of claims 27 to 32 wherein the reaction is effected at a temperature of from 0 to 1 OOC.

34. A process as claimed in claim 27 substantially as herein described.

35. A process for the preparation of adenosine triphosphate metal complexes substantially as herein described in Example 1.

36. Adenosine triphosphate metal complexes when prepared by a process as claimed in any one of claims 27 to 35.

37. Pharmaceutical compositions comprising as active ingredient an effective amount of at least one adenosine triphosphate metal complex in association with a pharmaceutical carrier or excipient.

38. Compositions as claimed in claim 37 in sterile form.

39. Compositions as claimed in claim 37 or claim 38 presented in the form of tablets, pills, capsules, vials or ampoules.

40. Compositions as claimed in any one of claims 37 to 39 in dosage unit form.

41. Compositions as claimed in claim 40 wherein each dosage unit contains from 1 to 150 mg of active ingredient.

42. Compositions as claimed in claim 41 wherein each dosage unit contains from 1 to 100 mg of active ingredient.

43. Pharmaceutical compositions as claimed in any one of claims 37 to 42 wherein the active ingredient comprises adenosine triphosphate cobalt complex.

44. Pharmaceutical compositions substantially as herein described.

45. Pharmaceutical compositions substantially as herein described in Example

2.

46. Adenosine triphosphate metal complexes for use in the treatment of cerebrovascular and cerebrosclerotic diseases, and/or of osteoporesis and cysts in bones.

47. Each and every novel complex, process and composition herein disclosed.

New Claims or Amendments to Claims filed on 9 Nov 1979 Superseded claim 1 New or Amended Claims: