WO1995019173A1

WO1995019173A1 - Pharmaceutical compositions

Info

Publication number: WO1995019173A1
Application number: PCT/HU1995/000001
Authority: WO
Inventors: András Egyed; László GAÁL; Gábor Gigler; Dóra Groó; István Gyertyán; Annamária SIMÓ; Katalin Szemerédi; Attila MÁNDI
Original assignee: EGIS Gyógyszergyár Rt.
Priority date: 1994-01-14
Filing date: 1995-01-13
Publication date: 1995-07-20
Also published as: AU1544195A; HU9400096D0; HUT71408A

Abstract

The invention relates to pharmaceutical compositions containing papaverine or pharmaceutically acceptable acid addition salts thereof, which are useful for the treatment or prevention of diseases associated with the glutamatergic neurotransmission. Such diseases are e.g. the transitional ischaemic attack, cerebral or spinal embolism, thrombosis, brain infarction, anoplexy, arteriosclerosis; various degenerative disorders of the central nervous system, such as sclerosis multiplex, Parkinson's disease, Huntington's disease or dementiae of Alzheimer or other origin. Papaverine is effective also against the sequels of metabolic origin, particularly diabetes mellitus or Wilson's syndrome. The invention furthermore relates to the preparation of the above compositions as well as the use of papaverine or pharmaceutically acceptable acid addition salts thereof for the treatment and prevention of the above diseases.

Description

PHARMACEUTICAL COMPOSITIONS

The invention relates to pharmaceutical compositions containing papaver- ine or pharmaceutically acceptable acid addition salts thereof, which are useful for the treatment or prevention of diseases associated with the glutamatergic neurotransmission.

Furthermore, the invention relates to the preparation of these composi¬ tions as well as the use of papaverine or pharmaceutically acceptable acid addition salts thereof for the treatment or prevention of diseases associated with the glutamatergic neurotransmission.

Papaverine, isolated by Merck in 1848, is a known alkaloid of opium. Nowadays papaverine is synthetically prepared. The water-soluble hydrochlo- ride of papaverine is a spasmolytic being used also at present. Indications of papaverine include e.g. spasms of the smooth muscles, arteriosclerosis obliter- ans and pulmonal embolism. Therapeutical doses of papaverine do not induce central nervous system effects. Higher doses elicit a mild sedative and weak analgesic action, however, these are practically not significant. After parenteral administration, it is well absorbed and it is effective in a dose of 50 to 100 mg [Jόzsef Knoll: Gyόgyszertan ("Pharmacology") in Hungarian, Edn. of Medicina, page 372, Budapest (1983)]. Papaverine hydrochloride is commercially avail¬ able in Hungary in the form of tablets or injectable compositions for the treat¬ ment of smooth muscle spasms (colics of bile, kidney and ureter stone, gastro¬ intestinal spasms, bronchial spasm, arterioclerosis obliterans, arterial and pul- monal embolism, cerebral ischaemia [see: "utmutato a gyόgyszerkeszitmenyek rendelesere ("Guide for the Prescription of Pharmaceutical Compositions" in Hungarian, Edn. of Medicina, page 521, Budapest (1988)]. Surprisingly, it has been found that papaverine very effectively inhibits in vitro the transmission mediated by the glutamatergic (i.e. non-NMDA) recep¬ tors. Furthermore, it has been found that papaverine binds as a noncompetitive antagonist to the receptor. By supplementing the data of the above in vitro measurements with pharmacological investigations it has been recognized that papaverine proved to be effective in the treatment of anoxia and histotoxic hy- poxia caused by cardiac arrest. These results prove the neuroprotective effect of papaverine.

The novel indications of papaverine recognized according to the present invention were demonstrated by the tests described hereinafter. In these tests the following compounds were used.

Compound to be tested: papaverine hydrochloride

Reference drugs:

GYKI-52466, -chemically: -1 -(4-aminophenyl)-4-methyl-7, 8-methylene- dioxy-5H-2,3-benzodiazepine;

CNQX:-6-cyano-7-nitroquinoxaline-2,3-dione;

NBQX:-2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f]quinoxaline.

Abbreviations used in the specification and in the biological tests are:

NMDA: N-methyl-D-aspartate; AMPA: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate.

1. Effect on the glutamatergic transmission on rat hippocampus slices

The effect of papaverine on the glutamatergic neurotransmission was in¬ vestigated on hippocampal brain slices by using the traditional method (Tarnawa et al, 1992). From the rat brains (a group consisted of 21 animals) transversal slices of 400 μm in thickness were prepared and kept living in an interface type chamber under artificial conditions mimicking the physiological conditions. After stimulating the Shaffer's collaterals the field potentials induced were led off from the pyramidal cells of the CA1 region. The neurotransmitter of this nervous path is glutamate and the transmission proceeds mainly through the AMPA receptors. The known AMPA antagonists (GYKI-52466, CNQX, NBQX) inhibit the field potentials in a concentration-dependent manner. Table I illustrates the results obtained with papaverine and GYKI-52466, respectively. The column of "inhibition%" shows the maximum effect achieved in the given concentration; the next column indicates the time of onset of the maximum ef¬ fect whereas values of the last column represent the time until the recovery of animals from effect of the compound.

Table I

Compound Concentra¬ Inhibition Time of Time of tested tion, μm % onset recovery min min

Papaverine 10 100 163 90

20 100 74 100

40 100 40 92

80 100 25 103

GYKI-52466 10 4.2 >60

20 27.5 >60

40 84.2 >60

»120 (70% inhibition)

The effectivity of papaverine is much stronger than that of GYKI-52466. Papaverine exerts an effect of 100% even in the lowest concentration used. Thus, its concentration inducing 50% inhibition is certainly significantly lower than 10 μM whereas this value is 26 μM for GYKI-52466. This effectivity is at least of the same order as that of NBQX found to be most effective up to now; however, according to the results of receptor binding papaverine acts at a site different from the AMPA receptor which is the binding site for NBQX.

The glutamatergic transmission-inhibiting action of papaverine is com¬ pletely reversible and does not involve any toxic property. Thus, it exceeds GYKI-52466 in this aspect, too.

2. Investigation of the receptor binding

The NMDA and non-NMDA receptor bindings were studied on mem¬ branes prepared from rat brain. The synaptic membrane fraction used in this measurement was obtained from the crude mitochondrial fraction of rat brain by centrifuging after a hyposmotic shock. In order to remove the endogenic ligand, this fraction was treated by several thawings and addition of Triton X-100 and again washed. The separation of the bound ligand from the free one was carried out by filtering in a Cell Harvester equipment.

The NMDA receptor binding was measured by using tritiated MK-801, a non-competitive antagonist ligand in the presence of glutamate and glycine in 5 mM Tris HCI buffer (pH = 7.4). The concentration of the ligand was 2 nM in a final volume of 0.5 ml. The incubation was carried out at room temperature for

30 minutes.

The AMPA receptor binding was measured by using tritiated AMPA in the presence of KSCN, a caotropic compound. The concentration of ligand was 10 nM in a final volume of 0.5 ml. The incubation was carried out at 4°C for 60 minutes. The kainate receptor binding was determined by using tritiated kainic acid in the presence of 2 mM CaCl2. The concentration of the ligand was 2 nM; the incubation was carried out at 4°C for 60 minutes.

At 10" 5 M concentration papaverine has no effect on the kainate and MK- 801 receptors and weakly blocks (about by 25%) the AMPA receptors. GYKI- 52466 has no influence on any of these receptors.

References:

- A. C. Foster and E. H. F. Wong: Br. J. Pharmac. 91., pages 403 to 409 (1987)

- D. E. Murphy, E. W. Snowhill and M. Williams: Neurochem. Res. 12, pages 775 to 782 (1987)

- T. Honore, J. Drejer and M. Nielsen: Neurosci. Let. 65, pages 47 to 52 (1986).

In the animal experimental models of neuroprotection, NBQX and GYKI- 52466 are effective (Smith and Meldrum, 1992); they exert an anticonvulsive effect in various models of epilepsy (Chapman et al., 1991). In several experi¬ mental models GYKI-52466 as a noncompetitive antagonist surpasses the ef¬ fect of NBQX (Yamaguchi et al., 1991). In opposition to the competitive block- ade, the noncompetitive blockade cannot be counterbalanced by the supple¬ mentary introduction of an agonist. Thus, a noncompetitive antagonist can be used at a lower level (concentration). The results of the receptor binding ex¬ periments described above indicate the noncompetitive character of papaverine which may be advantageous in comparison to that of NBQX type compounds. References:

- A. G. Chapman, S. E. Smith and B. S. Meldrum: Epilepsy Res. 9, pages 92 to 96 (1991)

- E. LePeillet, E. Arvin, c. Moncada and B. S. Meldrum: Eur. J. Neurosci, 4, Suppl. page 1068 (1991)

- S. E. Smith and B S. Meldrum: Stroke 23, pages 861 to 864 (1992)

- 1. Tarnawa, P. Molnar, L. Gaal and F. Andrasi: Acta Physiol. Hung. 79, pages 163 to 169 (1992)

- S. Yamaguchi, S. D. Donevan and M. A. Rogawski: Epilepsy Res. (in press).

3. Effect on the histotoxic hypoxia

Male Charles River Wistar rats (10 animals in each group) weighing 180 to 220 g each were used in these experiments. Histotoxic hypoxia was induced by the intravenous administration of 5.0 mg/kg of KCN. During the experiment, the time from the administration of KCN up to the last gasp of the animals was measured. Papaverine was intraperitoneally (i.p.) administered in distilled water solution 30 minutes before starting the experiment (Moore et al., 1986; Berga et al, 1986).

The last gasp of animals not treated with papaverine occurred by 37.6+0.9 seconds following the administration of KCN. The prolongation of the survival time (dST) by various doses of papaverine as well as the statistical significance level (P) are shown in Table II.

Table II Dose (mg/kg i.p.) dST (sec) P

10 2.4 0.110 20 4.1 0.035 40 6.8 0.026

80 8.1 0.008

120 6.3 0.001

According to the above results papaverine prolongs the time lasting up to the last gasp. This effect is even at a dose of 20 mg/kg statistically significant.

In the tissue hypoxia induced by KCN, the toxicity of cyanide ions is a consequence of the inhibition of cytochrom oxidase enzyme and subsequent injury of the cellular metabolism, which leads to abolishment of the cellular in- tegrity. The gasps are regulated by the neurons of the respiratory centre. Thus, the time of gasp is prolonged if the neuronal function is protected by a sub¬ stance for a certain period (Wauquier et al., 1985). It has been proven by this experiment that papaverine gets into the brain and exerts there a neuroprotec- tive effect. Since cyanide ions act directly on the cytochrom oxidase of brain cells, the condition of blood circulation has no influence on the result of this ex¬ periment.

References:

- R. Berga, P. R. Beckett, D. J. Roberts, J. Lienas and R. Massingham: Arzneim.-Forsch. [Drug Res. 36, pages 1314 to 1320 (1986) - S. J. Moore, J. C. Norris, I. K. Ho and A. S. Hume: Toxicol. Appl. Phar¬ macol. 82, pages 40 to 44 (1986)

- A. Wauquier, G. Clinke, D. Ashton, M. DeRyck, J. Fransen and G. Van Clemen: Drug Dev. Res. 8 (1986)

4. Investigation of neuronal injuries induced by transitional distur- bances of the cerebral blood supply

Male Mongolian gerbils weighing 60 to 70 g each were used in these ex¬ periments. Before the surgical intervention the animals were intraperitoneally (i.p.) treated with 10 mg/kg of papaverine for 4 days. In the 4th day a transi¬ tional insufficiency of cerebral blood supply (transitional cerebral ischaemia) was induced by the bilateral ligation of carotic arteries. Four days later the ani¬ mals were killed and microscopical sections were prepared from their brain by the silver impregnation method. In our experiments the necrosis of cortical and hippocampal neurons was diminished by papaverine.

In the cortical area papaverine exerts its effect in a lower dose than GYKI- 52466. In addition, GYKI-52466 has no protective effect in the hippocampus. References:

- M. J. Sheardown et al.: In: Pharmacology of Cerebral Ischaemia, pages 245 to 253 (1990)

- T. Freund et al.: Exp. Neurol. 108, pages 251 to 260 (1990).

Thus, the invention relates to a pharmaceutical composition for the treat- ment or prevention of diseases associated with the glutamatergic neurotrans¬ mission, which comprises papaverine or a pharmaceutically acceptable acid addition salt thereof together with pharmaceutically acceptable inert, solid or liquid carriers.

Furthermore, the invention relates to the preparation of pharmaceutical compositions useful for the treatment or prevention of diseases associated with the glutamatergic neurotransmission, which comprises mixing papaverine or a pharmaceutically acceptable acid addition salt thereof with pharmaceutically acceptable inert, solid or liquid carriers and transforming the mixture obtained to a galenical form. The invention relates also to the use of papaverine or its pharmaceutically acceptable acid addition salts for the treatment or prevention of diseases asso¬ ciated with the glutamatergic (neurotamatergic) neurotransmission.

The salts of papaverine formed with pharmaceutically acceptable acids may be addition salts of inorganic acids (such as hydrogen halides, e.g. hydro- chloric acid or hydrogen bromide; sulfuric acid or phosphoric acid); or addition salts with organic acids (e.g. formic, acetic, propionic, lactic, malic, fumaric, maleic, malonic, citric or nicotinic acid and the like). Papaverine hydrochloride is the preferred salt.

The pharmaceutical compositions of the invention which contain papaver- ine or a pharmaceutically acceptable acid addition salt thereof, are useful for the treatment or prevention of diseases associated with the injury of neurons of the central nervous system. Thus, according to the invention, these pharma- ceutical compositions can be utilized e.g. for the treatment or prevention of the following disorders: Neuronal injuries of various origin occurring as sequels of hypoxic or haemorrhagic states (e.g. transitional ischaemic attack, cerebral or spinal embolism, thrombosis, brain infarction, apoplexy (stroke), arteriosclerosis and the like); various degenerative neurological clinic pictures (e.g. sclerosis multiplex, amyotrophic lateral sclerosis, Parkinson's disease, Huntington's dis¬ ease, dementia of Alzheimer or other origin etc.); alleviation of the sequels of cerebral and spinal injuries of mechanical origin (such as traumas, parturitive traumas or accidental traumas) or after-effects of comatose conditions occur- ring therein; injuries of the central nervous system developing as sequels of metabolic disorders (such as diabetes mellitus or Wilsons's syndrome); various types of epileptic clinical pictures (e.g. clinical patterns of Jackson's type, grand mal); or chirurgical pre-medication.

According to the invention papaverine and pharmaceutically acceptable acid addition salts thereof are useful also for the protection against the sequels of anoxia or histotoxic hypoxia induced by cardiac arrest.

When used as neuroprotective agents, papaverine or the pharmaceuti¬ cally acceptable acid addition salts thereof may usually be administered in daily doses between about 1 mg and about 1000 mg, preferably about 10 to 150 mg. The pharmaceutical compositions according to the invention may contain the usual components and can be prepared by using known methods of the drug manufacture.

The pharmaceutical compositions of the invention can be formulated for oral, parenteral or rectal use, in the form of nasal drops or spray or for admini- stration by inhalation. For oral use, the compositions may preferably be in the form of tablets, coated tablets, dragees, capsules, syrups or powders. These compositions may contain commonly used pharmaceutical carriers and/or auxil¬ iaries, e.g. lactose, starch, magnesium stearate, sodium citrate, calcium carbon ate, dicalcium phosphate, starch derivatives (e.g. carboxymethyl-starch, sodium starch glycollate), silicic acid, binding substances (e.g. polyvinylpyrrolidone and the like) or sliding agents (e.g. magnesium stearate or the like) are useful for this purpose. It is suitable to prepare tablets containing 40 to 100 mg of active ingredient each.

The compositions intended for oral administration may be also aqueous suspensions, solutions or elixirs. These compositions may comprise e.g. water, ethanol, propylene glycol or glycerol as diluents. Furthermore, the usual addi¬ tives (e.g. colouring, emulsifying and stabilizing agents, such as e.g. methyl p- hydroxybenzoate) may also be ingredients of the compositions.

Tablets can be prepared by dry or wet granulating processes. Dragees may be prepared by forming the dragee core and then covering it with a suit- able coat in a manner known per se. In order to prepare capsules, an appro¬ priate mixture is firstly prepared, which is then filled into hard or soft gelatine capsules.

Suppositories intended for rectal administration usually contain 40 to 100 mg of active ingredient. Suppositories are prepared by uniformly distributing the active ingredient in the molten suppository base (e.g. cocoa butter, Witepsol H 15 or the like), filling the melt into proper moulds, cooling it and finally, packing the suppositories in aluminum or tin foil.

Injectable compositions intended for parenteral use may be administered in intravenous, intraperitoneal or subcutaneous route. The active agent content of the injectable solutions usually varies between 40 mg/ml and 100 mg/ml. In¬ jectable solutions are commonly filled into ampoules of 1 or 2 ml volume. Par- enterally injectable solutions may preferably contain water, sesame oil peanut oil, aqueous propylene glycol or other pharmaceutically acceptable solvents as diluents. The use of aqueous solutions is preferred. When necessary, aqueous solutions can be buffered or isotonized by using sodium chloride or glucose in a known manner. If desired, the solutions obtained may be sterilized in a known way. In addition, infusion solutions and plasters may be used for parenteral administration.

The invention will be illustrated in detail by the following non-limiting Ex- amples. Example 1

Tablets containing the following ingredients are prepared by using known methods of drug manufacture.

Ingredients Amount, mg/tablet Papaverine hydrochloride 40

Magnesium stearate 4

Sodium starch glycollate 20

Lactose 80

Talc 20 Example 2

Injectable solutions containing 40 mg of papaverine hydrochloride per 1 ml of distilled water useful for injection purpose are prepared by using known methods of drug manufacture.

Example 3 Suppositories containing the following ingredients are prepared by using known methods of drug manufacture.

Ingredients Amount, mg/suppositorv

Papaverine hydrochloride 40

Witepsol H 15 200 Example 4

Ingredients Amount, mg/tablet

Papaverine hydrochloride 50

Lactose 100

Potato starch 100

Sodium amylopectin glycollate 8

Gelatine 2

Magnesium stearate 3

Example 5

Tablets containing the following ingredients are prepared by using known methods of drug manufacture. Ingredients Amount, mg/tablet

Papaverine hydrochloride 40

Stearin 1

Potato starch 20

Lactose 39

Claims

Claims:

1. Process for the preparation of a pharmaceutical composition for the treatment or prevention of diseases associated with the glutamatergic neuro- transmission, which comprises mixing papaverine or a pharmaceutically ac¬ ceptable acid addition salt thereof prepared in a manner known per se with pharmaceutically acceptable inert, solid or liquid carriers and transforming the mixture obtained to a galenic form.

2. A process as claimed in claim 1, which comprises preparing pharma- ceutical compositions for the treatment or prevention of diseases associated with the neuronal injury of the central nervous system.

3. A process as claimed in claim 2, which comprises preparing pharma¬ ceutical compositions for the protection against or alleviation of neuronal inju¬ ries occurring as sequels of hypoxic or haemorrhagic states of various origin. 4. A process as claimed in claim 3, which comprises preparing pharma¬ ceutical compositions for the treatment or prevention of transitional ischaemic attack, cerebral or spinal embolism, thrombosis, brain infarction, apoplexy or arteriosclerosis.

5. A process as claimed in claim 2, which comprises preparing pharma- ceutical compositions for the treatment or prevention of various degenerative neurological clinic pictures.

6. A process as claimed in claim 5, which comprises preparing pharma¬ ceutical compositions for the treatment of sclerosis multiplex, amyotrophic lat¬ eral sclerosis, Parkinson's disease, Huntington's disease or dementiae of Alz- heimer or other origin.

7. A process as claimed in claim 2, which comprises preparing pharma¬ ceutical compositions for the treatment of cerebral or spinal injuries of me¬ chanical origin, particularly traumas, parturitive traumas or accidents and coma¬ tose states occurring therein. 8. A process as claimed in claim 2, which comprises preparing pharma¬ ceutical compositions for the treatment or prevention of central nervous system injuries occurring as the sequels of metabolic disorders, particularly diabetes mellitus or Wilson's syndrome.

9. A process as claimed in claim 2, which comprises preparing pharma¬ ceutical compositions for the treatment or prevention of epileptic clinical pic- tures, particularly of Jacksonian type or grand mal or for use in the chirurgical premedication.

10. A process as claimed in claim 2, which comprises preparing pharma¬ ceutical compositions for the protection against the sequels of anoxia or histo¬ toxic hypoxia induced by cardiac arrest. 11. Pharmaceutical composition for the treatment or prevention of dis¬ eases associated with the glutamatergic neurotransmission, which comprises papaverine or a pharmaceutically acceptable acid addition salt thereof together with pharmaceutically acceptable inert, solid or liquid carriers.

12. A pharmaceutical composition as claimed in claim 11 for the treatment or prevention of diseases associated with the neuronal injury of the central nervous system.

13. A pharmaceutical composition as claimed in claim 12 for the protection against or alleviation of neuronal injuries occurring as the sequels of hypoxic or haemorrhagic states of various origin. 14. A pharmaceutical composition as claimed in claim 13 for the treatment or prevention of transitional ischaemic attack, cerebral or spinal embolism, thrombosis, brain infarction, apoplexy or arteriosclerosis.

15. A pharmaceutical composition as claimed in claim 12 for the treatment or prevention of various degenerative neurologic clinical pictures. 16. A pharmaceutical composition as claimed in claim 15 for the treatment of sclerosis multiplex, amyotrophic lateral sclerosis, Parkinson's disease Hunt¬ ington's disease or dementiae of Alzheimer or other origin.

17. A pharmaceutical composition as claimed in claim 12 for the treatment of cerebral or spinal injuries of mechanical origin, particularly traumas, parturi- tive traumas, accidents or comatose conditions occurring therein. 18. A pharmaceutical composition as claimed in claim 12 for the treatment or prevention of central nervous system injuries occurring as the sequels of metabolic disorders, particulariy diabetes mellitus or Wilson's syndrome. t9. A pharmaceutical composition as claimed in claim 12 for the treatment or prevention of epilectic clinical pictures, particularly of Jacksonian type or grand mal or for use in chirurgical premedication.

20. A pharmaceutical composition as claimed in claim 12 for the protection against the sequels of anoxia or histotoxic hypoxia induced by cardiac arrest.

21. A process as claimed in any of the claims 1 to 10, which comprises using papaverine hydrochloride as active ingredient.

22. A pharmaceutical composition as claimed in any of the claims 11 to 20, which comprises papaverine hydrochloride as active ingredient.

23. Use of papaverine or pharmaceutically acceptable acid addition salts thereof for the treatment or prevention of diseases associated with the glutama- tergic neurotransmission.

24. Use of papaverine or pharmaceutically acceptable acid addition salts thereof for the treatment or prevention of diseases defined in any of the claims 2 to 10.