SE443716B - SETTING TO PREPARE A PHARMACEUTICAL PREPARATION FOR INHIBITING BLOOD SPOT SEGREGATION CONTAINING PROSTACYCLINE OR DIHYDROPROSTACYCLINE AND A PHOSPHODIESTERATION INHIBITOR - Google Patents

Description

7805707-2 10 15 20 25 30 35 2 of postoperative thrombosis, for the promotion of patency of vascular grafts after surgery and for the treatment of arteriosclerosis complications and conditions such as arteriosclerosis, blood coagulation defects due to lipemia and other clinical conditions where the underlying etiology is associated with lipidobalance or hyperlipidemia.

The active compounds are particularly useful as additives to blood, blood products, blood substitutes and other fluids used in artificial extra-corporeal circulation and perfusion of an isolated body part, such as a limb or organ, whether or not attached to the original. the body, loosened and preserved or prepared for transplantation, or attached to a new body. During these circulations and perfusions, aggregated platelets tend to block blood vessels and parts of the circulatory system. This blockage can be avoided by the presence of the active compounds.

For this purpose, the compound may be administered gradually or in a single portion or multiple portions to the circulating blood, to the blood of the donor animal, to the perfused body part, which is detached or attached, to the recipient, or to two or all of these in a total stationary dose of from 0.001 to 10 mg / l of circulating fluid. It is advisable to use these compounds in laboratory animals, such as cats, dogs, rabbits, monkeys and rats, for these purposes to develop new methods and techniques for organ or limb transplantation.

In addition to the effects described above, the active compounds also show a vasodilatory effect on blood vessels. It often occurs when it would be desirable to be able to utilize the other effects described above (in particular the anti-aggregatory effects) while the vasodilatory effect of the active compounds is simultaneously suppressed or eliminated, for example in the treatment or prevention of myocardial infarctions and when used as an adjunct in extra-corporeal circulation. The present invention involves tube excitation or avoidance of this vasodilation complication.

It has now surprisingly been discovered that the anti-aggregatory effects of the active compounds, but not the vasodilatory effects, are potentiated by chemical compounds which show a special pharmacological activity. This finding provides an opportunity to distinguish the anti-aggregatory effects of the active compounds from their vasodilatory effects.

Accordingly, the present invention provides a method of preparing a pharmaceutical formulation, which formulation comprises: (a) Prostacyclin, dihydroprostacyclin or a pharmaceutically acceptable salt of any of these, (b) a phosphodiesterase inhibitor and optionally (c) a pharmaceutically acceptable carrier .

The preparation promotes an anti-aggregatory effect on platelets.

The combined use of an active compound and a phosphodiesterase inhibitor lowers the anti-aggregatory threshold level (the minimum amount of active compound required to produce anti-aggregatory effects) without significantly affecting the vasodilatory threshold level.

By administering an active compound at a concentration which is below its vasodilatory threshold level but which is above the new, lower anti-aggregatory threshold level, together with a phosphodiesterase inhibitor, anti-aggregatory effects can be achieved while vasodilatory effects are not achieved.

It is known that the active compounds can stimulate the production of cyclic AMP in platelets and it has been suggested that the anti-aggregatory effects of the active compounds may contribute to the formation of cyclic AMP. Phosphodiesterase inhibitors inhibit the degradation of cyclic AMF and therefore it may be possible for the potentiation of the anti-aggregatory effects to occur, for in the presence of a phosphodiesterase inhibitor a suitable, anti-aggregatory, concentration of cyclic AMF may occur. AMP is obtained with a lower concentration of active compound.

The mechanism by which the active compounds manifest their vasodilatory effects is not known, although a relationship between the concentration of cyclic AMP and vasodilatory activity has been suggested for other compounds. Our discovery that the vasodilatory effects of the active compounds used in the present invention are not potentiated by phosphodiesterase inhibitors suggests that cyclic AMP is not involved in the vasodilatory effect of the active compounds.

Suitable phosphodiesterase inhibitors for use in potentiating the anti-aggregatory effects of the active compounds include: Xanthine derivatives such as: Theophylline [3,7-dihydro-1,3-dimethyl-1H-purine-2,6-dione], and salts thereof. 3-isobutyl-1-methyl-xanthine, Caffeine [3,7-dihydro-1,3,7-trimethyl-1H-purine-2,6-dione] and salts thereof; and Aminophylline [adduct of Theophylline and 1,2-ethanediamine (2: 1)].

Phosphodiesterase inhibitors that can be used in the preparation of the present invention other than xanthine derivatives include, as such or as pharmaceutically acceptable salts: (a) Isoquinoline derivatives, for example: Papaverine [11 (3,4-dimethoxyphenyl) methyl] -6, 7-dimethoxyisoquinoline] and salts thereof; and 6,7-diethoxy-1- (4,5-diethoxybenzyl) isoquinoline or its salts, for example its hydrochloride; (B) Derivatives of pyrimido [5,4-dylpyrimidine, for example Dipyridamol [2,2 ', 2 ", 2"' - (4,8-dipiperidino-pyrimido- [5,4 -d] pyrimidine-2,6-diyldinitrilo) tetraethanol) and its salts; 2,2 L, 2-32 "', - [[4- (bipiperinyl) pyrimidote, 4-alpyrimidine-2,6-diyl dinitrilo] tetrakisethanol and its salts, and 2,4,6-tri-4-morpholinylpyrimido [5,4-d] pyrimidine and its salts (c) Derivatives of thieno [3,2-d] pyrimidine, for example N- [4 - (4-morpholinyl) thieno] 3,2-d] pyrimidin-z-yl] -1,2-ethanediamine (d) Derivatives of pyrazolo [3 ', 4': 2,3] pyrido [4,5-b ] [1,5] -benzodiazepin-6- (3H) -one, for example 3-ethyl-7,12-dihydro-7,2-dimethylpyrazolo [4 ', 3': 5,6] -pyridoI4,3- b] - [1,5] benzodiazepin-6- (3H) -one; 3-ethyl-7,12-dihydro-9-methoxy-7,2-dimethyl-pyrazolo [3 ', 4': 2,3 ] pyrido [4,5-b] [1,5] benzodiazepin-6- (3H) -one, and 10-chloro-3-ethyl-7,12-dimethyl-7,12-dihydro-pyrazolo [4 ', 3 ': 5,6] pyrido [4,3-b] [1,5] benzodiazepin-6- (3H) -one; (e) Derivatives of 1H- or ZH-pyrazolo [3,4-b] pyridine, for example 4- (butylamino) -1-ethyl-1H-pyrazo lo [3,4-b] pyridine-5-carboxylic acid ethyl ester; 4- (butylamino) -1H-pyrazolo [3,4-b] pyridine-6-carboxylic acid ethyl ester; 4-chloro-1-ethyl-3-methyl-1H-pyrazolo [3,4-b] pyridine-5-acetonitrile; 1-ethyl-4- (isopropylidenehydrazino) -3-methyl-1H-pyrazolo [3,4-b] pyridine-5-carboxylic acid ethyl ester or its salts such as its hydrochloride semihydrate; and 2-methyl-6-phenyl- 4- (1-piperidinyl) -2H-pyrazolo [3,4-b] pyridine or its salts, for example its hydrochloride. (f) Derivatives of sH-furois fi x-e] pyrazoiols, 4-bipyrimin-s-one, for example 4- (butylamino) -1-ethyl-1,7-dihydro-7-hydroxy-5H-furo- - [3, 4-e] pyrazolo [3,4-b] pyridin-5-one; and 6805707-2 6 (g) Derivatives of 1 (2H) -naphthalenone, for example 2 [(dimethylamino) methyl] -3,4-dihydro-7-methoxy-1 (2H) -naphthalenone or its salts such as its 1: 1 hydrochloride.

The amount of active compound required for therapeutic effect varies not only with the particular compound and mode of administration but also with the particular phosphodiesterase inhibitor used and its amount, i.e., with the degree of potentiation that occurs. However, the amount of active compound will be below the amount required to produce a given therapeutic effect in the absence of the potentiator and thus also below the level which would produce a significant vasodilatory effect.

In general, a suitable dose of active compound in the absence of a potentiator ranges from 0.01 to 200 pg / kg of the recipient's body weight. In the present invention where a phosphodiesterase inhibitor is used, a suitable dose of active compound is usually less than, or about, half the dose in the absence of potentiator, for example in a range from 0.002 to 100 pg per kg of body weight.

The present invention provides a method of preparing a formulation which comprises mixing an active compound and a phosphodiesterase inhibitor.

In general, the active compound and the phosphodiesterase inhibitor are administered in a weight ratio of one part of the active compound to from 1 to 200 parts of the phosphodiesterase inhibitor.

Although it is possible to administer an active compound and a phosphodiesterase inhibitor as crude chemicals, it is preferred to present them as one or more pharmaceutical preparations.

A unit dose of a preparation may contain from 0.1 mg to 20 mg, for example from 0.2 to 16 mg of the active compound and from 100 to 400 mg, for example from 250 to 350 mg of the phosphodiesterase inhibitor. When the phosphodiesterase inhibitor is incorporated into a formulation prepared according to the invention, the formulation comprises, for both veterinary and human medical use, an active compound and a phosphodiesterase inhibitor together with one or more acceptable carriers therefor and optionally a other therapeutic ingredient or other therapeutic ingredients. The carrier must be "acceptable" in the sense that it must be compatible with the other ingredients of the preparation and must not be harmful to the recipient.

The formulations include those suitable for oral, rectal, vaginal or parenteral (including subcutaneous, intramuscular or intravenous injection or infusion) or intrapulmonary administration, although the most appropriate mode of administration in any case depends on the active compound.

The formulations are conveniently presented in unit dosage form and may be prepared by methods known in the art of pharmacy. All such methods include the step of bringing into association the active compound and the phosphodiesterase inhibitor with the carrier which is constituted by one or more accessory ingredients. In general, the formulations are prepared by bringing the active compound and the phosphodiesterase inhibitor into a homogeneous and intimate compound, for example by mixing, with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, forming the formulation into the desired product. .

Formulations suitable for oral administration prepared according to the invention may be presented as stand-alone units such as a capsule, lozenges or tablets, each containing a predetermined amount of the active compound and the phosphodiesterase inhibitor in the form of a powder or granules; or as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in-water emulsion or a water-in-oil-liquid emulsion. A preparation suitable for parenteral administration, prepared according to the invention, may be presented in an ampoule for receiving a defined amount of liquid to make a solution for infusion.

An example of such a parenteral preparation is a lyophilized residue of a solution of the active compound, the phosphodiesterase inhibitor and a glycine buffer of pH 10.5.

A tablet may be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active compound and the phosphodiesterase inhibitor in a free-flowing form, such as a powder or granules, optionally mixed with a binder, lubricant, pharmaceutically inert diluent, surfactant or dispersant. Molded tablets may be prepared by molding in a suitable machine a mixture of the powdered active compound and a phosphodiesterase inhibitor, together with a suitable carrier moistened with a pharmaceutically inert liquid diluent.

Preparations for rectal administration can be presented as a suppository with a conventional carrier, such as cocoa butter.

Formulations suitable for vaginal administration may be presented as a pessary, a cream, a paste or a spray formulation containing, in addition to the active compound and the phosphodiesterase inhibitor, such carriers as are known in the art to be suitable.

Formulations suitable for parenteral administration suitably comprise a sterile, aqueous preparation of the active compound and the phosphodiesterase inhibitor, which is preferably isotonic with the recipient's blood.

Since the active compound can be absorbed through the skin into the blood of the recipient, it can be administered topically in a topical preparation together with the phosphodiesterase inhibitor if it can be absorbed through the skin into the blood. For the treatment of injuries in or near the skin, the topical administration of the preparation is particularly suitable.

It is to be understood that the formulations prepared according to the invention may comprise, in addition to the above-mentioned ingredients, one or more additional ingredients, such as a diluent, a buffer, a lubricant or a preservative (including an antioxidant).

Thus, in accordance with the present invention, there is provided: A method of preparing a pharmaceutical preparation for inhibiting platelet aggregation, characterized by mixing (a) prostacyclin, dihydroprostacyclin or a pharmaceutically acceptable salt thereof, (b) a phosphodiesterase inhibitor and optionally (c) a pharmaceutically acceptable carrier, the components (a) and (b) being used in a weight ratio of 1: 1 to 1: 200.

The following examples illustrate the invention.

EXAMPLE 1 Thrombi were induced in rabbit carotid artery by the method described by R.G. Herrman and W.B. Halefield, Effects of antithrombotic drugs in in vivo experimental thrombosis in S. Sherry and A. Scriabine (eds), Platelets and ïhnm fi xsis, University Park Press, London, l974.

Following thrombus infusion, intravenous (iv) infused either an active compound, phosphodiesterase inhibitor alone or a combination of an active compound and phosphodiesterase inhibitor for a period of time (30 minutes for administration of a single compound or 30 minutes from the start of the infusion of the active compound). if a compound was used) and the carotid artery was cut and opened and the size of the thrombus is valued at a beämnhmßä fifl a (+ to +++). The absence of a tornado was valued as a -. In the case where a composition of active compound and phosphodiesterase inhibitor was used, i.v. the infusion of the phosphodiesterase inhibitor 5 minutes before the infusion of the active compound. The active compound was infused at a wide sub-threshold concentration (ie at a concentration which, when used alone, was insufficient to exhibit anti-aggregatory effects).

The results obtained are shown in Table 1. The animals' blood pressure was monitored throughout the experiments. No effect on blood pressure was observed. 10 78057Û7 * 2 ll TABLE l Active compound and concentrates ~ Posfodiesterase inhibitors Thrombation and concentration sterleK_ Artery Prostacyclin None +++ L.C. at 3-isobutyl-1-methyl-xanthine + R, C_ 50 ng / min.kg at 10 pg / min.kg Prostacyclin None ++ L.C. at 3-isobutyl-1-methyl-xanthine - R.C. 25 ng / min.kg at 50 pg / min.kg Prostacyclin No +++ L, C, at Theophylline at 25 ng / min.kg 125 pg / min.kg - R.C.

Prostacyclin None + x R.C. at Theophylline at 10 ng / min.kg 125 pg / min.kg - L.C.

Prostacyclin None ++ L.C. at Theophylline at 25 ng / min.kg 125 pg / min.kg + R.C.

Prostacyclin None ++ L, C. at Theophylline at + R.C. 10 ng / min.kg 200 pg / min.kg None 'None +++ L.C.

Prostacyclin Theophylline + R.C. at at 10 ng / min.kg 200 pg / min.kg None None +++ R.C.

Theophylline at +++ L.C.

X 1 h 15 min elapsed before infusion of active compound was started. 200 pg / min.kg 7805707-2 Active compound 12 TABLE 1 (continued) and concentrates- Phosphodiesterase inhibitors Thrombosis and concentration size Arterial Dihydroprosta- None +++ cyclin at 50 ng / min.kg Dihydroprosta- None + (2 ex - cyclin at peri- 100 ng / min.kg ment) Dihydroprosta- None - cyclin at 250 ng / min.kg Dihydroprosta- None ++ LC cyclin at Theophylline at 50 ng / min.kg 200 pg / min.kg ++ R.C.

Dihydroprosta- None +++ L.C. cyclin at Theophylline at 50 ng / min.kg 200 Pg / min.kg + R.C.

Dihydroprosta- No +++ L.C cyclin at Theophylline at - R.C. 50 ng / min.kg 200 pg / min.kg 10 15 25 25 25 35 7805707-2 13 EXAMPLE 2 Rabbits were anesthetized with pentobartitone (30 mg / kg iv) and heparin (2500 IU / kg iv) was injected immediately before withdrawal. of blood. Arterial blood from a carotid artery of the rabbits was taken continuously at 3 ml / min by means of a roller pump and used to overheat collagen strips, which were excised from another rabbit's Achilles tendon.

The tendons were suspended freely from auxotonic transducers (see Paton W.D.M. J. Physiol., 1957, lål, 35P), the output signals of which were due to the weight of the tendon and were displayed on a multi-channel recorder. Platelets, and possibly other cells, became stuck during the overflow of the tendons, thereby increasing their weight. Changes in weight due to platelet aggregation and / or adhesion were recorded as deviations of the pencils, which had previously been calibrated in milligrams. After shedding of the tendons, the blood was returned intravenously to the rabbit by gravity.

When the blood pouring over the tendons had begun, the tendon resemblance cell aggregates and increased in accumulated weight over a period of time from 35 to 40 minutes. Thereafter, there was no further increase in weight; the average weight gain was 93l15 mg; n = 19 (n is the number of experiments).

In the following experiments, the poured late strip strips, as above, were allowed to achieve a stable increase in weight due to cell deposition. Each subsequent weight loss then indicated disaggregation of cells. Prostacyclin sodium salt, (See Johnson, R.A., Lincoln, F.H., Thompson, J.L. Nidy, E.G., Mizsak, S.A., Axen, U. J. Am. Chem.

Soc., 1977, 22, 4182, was dissolved in Tris buffer (50 mM pH 7.4) and infused (0.1-1 ng / ml of blood for 3 minutes) into the blood where the tendons bathed. This induced disaggregation of cells, which were dose-dependent and reversible; 0.1 ng / ml of prostacyclin in the arterial blood induced 26: 3 mg (n = 10) disaggregation in arterial blood, and 1 ng / ml induced 86É6 mg (n = 6) disaggregation. 7805707-2 V1 10 15 20 p. 14 Dipyridamole was infused into the blood so that the tendons were shed and also induced a dose-dependent and reversible disaggregation.

Dipyridamole and prostacyclin were co-infused into the blood as the overlying theses and produced a clear synergism of their individual disaggregation effects.

The concentrations of prostacyclin-sodium and of dipyridamole used in the comparison are shown in Table 2, together with the disaggregations obtained, expressed as a percentage of the maximum disaggregation that could theoretically be obtained. (The difference in weight between the tendon with maximum aggregation and the initial weight of the tendon).

EXAMPLE 3 A lyophilized preparation was prepared by dissolving dihydroprostacyclin sodium salt (1 mg) and dipyridamole (300 mg) in aqueous solution for injections (European Pharmacopoeia), which contained a glycine buffer of glycine (0.025M) and sodium chloride (0.025M). The resulting solution was adjusted to pH 10.5 with sodium hydroxide, which formed part of the buffer, and lyophilized.

Shortly before use, the lyophilized material was reconstituted in Water for injections and then administered by infusion or perhaps by injection.

EXAMPLE 4 This example was performed as Example 3 except that 1 mg of dihydroprostacyclin sodium was replaced with 8 mg of dihydroprostacyclin sodium.

EXAMPLES 5 and 6 These were carried out as Examples 3 and 4, respectively, except that dihydroprostacyclin sodium was replaced by an equivalent amount by weight of prostacyclin sodium. 10 15 20 7805707-2 15 TABLE 2 Prostacyclin-% Disaggre- -Sodium- Qation of infusion, Fosíodieste- platelet-concentration-inhibitor aggregate in Number of tion ng / ml of and concentric-arterial Exper- tration blood in sem ment According to nipyridamole 1oï1 14 l Pg / ml a single dose 0.05 None 2 Dipyridamol 28i2 l pg / ml a single dose o, 1 None 1sï1 io Dipyridamol 38: 5 6 l pg / ml a single dose o, s None 22252 1o Dipyridamole 45i3 6 1 pg / ml single dose Formulations, as the present invention, tive skin diseases, such as psoriasis, wherein by proliferating as creams the formulations are presented in suitable form, or lotions, suitable for topical administration to the affected area and to nearby areas. prepared in the manner according to can be used in treatment

Claims (14)

7805707-2 10 15 20 25 30 16 PÄT EN TK RAV 1. A method of preparing a pharmaceutical preparation for inhibiting platelet aggregation, characterized by mixing (a) prostate - cyclin, dihydroprostacyclin or a pharmaceutically acceptable salt of any of these, (b) a phosphodiesterase inhibitor and optionally (c) a pharmaceutically acceptable carrier, wherein components (a) and (b) are used in a weight ratio of 1: 1 to 1: 200. 2. A method according to claim 1, characterized in that the pharmaceutically acceptable salt is prostacyclin sodium or dihydroprostacyclin sodium. 3. A method according to claim 1 or 2, characterized in that the phosphodiesterase inhibitor is a xanthine derivative. A method according to claim 3, wherein the xanthine derivative is 3-isobutyl-1-methylxanthine known as caffeine or a caffeine, or a pharmaceutically acceptable salt thereof. 5. A method according to claim 3, characterized in that the xanthine derivative is theophylline or aminophylline or a pharmaceutically acceptable salt of either of these. 6. A method according to claim 1 or 2, characterized in that the phosphodiesterase inhibitor is known - a derivative of isoquinoline, pyrimido [5,4-d] pyrimidine, thieno [3,2-d] pyrimidine, pyrazolo [3 ', 4': 2, 3] pyrido [4,5-b] - [1,5] benzodiazepin-6- (3H) -one, 1H- or 2H-pyrazolo [3,4-b] pyridine, 5H-furo [3,4-e] pyrazolo [3,4-b] pyridin-5-one, or 1- (ZH) -naphthalenone. A process according to claim 6, wherein the derivative is Papaverine 11-1 (3,4-b-dimethoxyphenyl) methyl] -6,7-dimethoxyisoquinoline]; 6,7-di-ethoxy-1- (4,5-diethoxybenzyl) isoquinoline or its hydrochloride} Dipyridamole [2,2 ', 2 ", 2"' - (4,8-ipipipricoline) 7 pyrimido [5,4-dipyrimidine-2,6-diyldinitrilo) tetraethanol]; 2 ', 2', 2 ", 2" '- [[4- (1-piperidinyl) pyrimido [5,4-d] pyrimidine--2,6-diyl] dinitrilol tetrakisethanol; 2,4,6-tri-4-morpholinylpyrimido [S, 4-d] pyrimidine; N- [4- (4-morpholinyl) thieno- [3,2-d] pyrimidin-2-yl] -1,2-ethanediamine; 3-ethyl-7,2-dihydro-7,12-dimethylpyrazolo [4 ', 3': 5,6] pyrido [4,3-b] - [1,5] -benzodiazepine-6- (3H) -on; 3-ethyl-7,12-dihydro-9-methoxy-7,2-dimethyl-pyrazolo [3 ', 4': 2,3] pyrido [4,5-b] [1,5] benzodiazepine 6- (3H) -one; 10-chloro-3-ethyl-7,2-dimethyl-7,12-dihydropyrazolo [4 ', 3': 5,6] pyrido [4,3-b] [1,5] benzodiazepine-6- ( 3H) ~ on; 4- (butylamino) -1-ethyl-1H-pyrazolo-1,3,4-b] pyridine-5-carboxylic acid ethyl ester; 4-chloro-1-ethyl-3-methyl-1H-pyrazolo [3,4-b] pyridine-5-acetonitrile; 1-ethyl-4- (isopropylidenehydrazino) -3-methyl-1H-pyrazolo [3,4-b] pyridine-5-carboxylic acid ethyl ester; zolo [3,4-b] pyridine-6-carboxylic acid ethyl ester; 2-methyl-6-phenyl-4- (1-piperidinyl) -2H-pyrazolo [3,4-b] pyridine; 4-Butylamino-1H-pyra-4- (butylamino) -1-ethyl-1,7-dihydro-7-hydroxy-5H-furo- - [3,4-e] pyrazolo [3,4-b] pyridine-5- on; or 2 [(dimethylamino) methyl] -3,4-dihydro-7-methoxy-1- (2H) -naphthalenone or a pharmaceutically acceptable salt of such a derivative. The method of claim 2, wherein the phosphodiesterase inhibitor is dipyridamole. k ä n n e t e c k n a t k ä n n e t e c k n a t 9. A method according to claim 1, wherein the carrier is a solid. 10. A method according to claim 1, characterized in that the carrier is a liquid. 11. A method according to claim 1, characterized in that the preparation is made in a form suitable for oral, parenteral, rectal, vaginal or intrapulmonary administration. 12. A method according to claim 11, wherein the preparation is made in a lyophilized form for parenteral administration in an ampoule for receiving a certain amount of liquid for reconstitution of a solution known as a 78057Û7-2 18 for infusion. 13. A method according to claim 9, characterized in that the preparation is made in tablet form. 14. A method according to any one of claims 9-13, characterized in that the preparation is made in unit Lfl dosage form.