HU190513B - Process for producing of new preparates consisting of hydrogenised ergot-alcaloids and hepharine with therapeutical effect - Google Patents

Description

FIELD OF THE INVENTION The present invention relates to a novel therapeutic composition comprising a vasoconstrictive hydrogenated ergot alkaloid or a pharmaceutically acceptable acid addition salt thereof, and an Ids molecule, heparin or a pharmaceutically acceptable salt thereof 45 853-2703 / PT. The composition of the invention is thromboses

especially for the prevention of postoperative thromboses.

Co-administration of an average molecular weight of 14,000 to 19,000 molecules of natural heparin with a hydrogenated ergotalkaloid, particularly dihydroergotamine, with vasoconstrictive effects is known to have particular benefits in preventing thrombosis (see, for example, German Patent Publication No. 2,554,533). . Accordingly, co-therapy with heparin and dihydroergotamine is a valuable and generally accepted form of treatment for patients at risk of thrombosis.

especially as a result of surgeries. The administration is usually parenteral, that is, by intravenous injection. For example, compositions known in the literature are more stable than solutions commonly used in the above-mentioned indication (U.S. Patent 4,402,929). Dyen preparations are commercially available. Surprisingly, the present invention has been found to provide particularly good results in the prevention of postoperative thrombosis by using, for example, the heparin component of the combination described in German Patent Publication No. 2,554,533, which has an average molecular weight of natural. low relative to the molecular weight of heparin. In particular, it has been found that, when used with low molecular weight heparin in combination with hydrogenated ergot alkaloids, particularly dihydroergotamine, the duration of action against thrombosis, and thus the potential for therapeutic use, is dramatically lower than with known natural heparin formulations. is improved because the daily dose of heparin and the hydrogenated ergot alkaloid component needed to effectively prevent thrombosis can be reduced accordingly.

Accordingly, the present invention relates to a process for the preparation of a therapeutic composition which substantially reduces the dose of a combination of a vasoconstrictive hydrogenated ergot alkaloid and heparin for the treatment of thrombosis. Because the combination is conveniently administered by injection, the therapeutic composition of the present invention can alleviate the patient's anxiety by simply administering the composition once daily. This is especially important for patients recovering from major surgery.

SUMMARY OF THE INVENTION The present invention relates to a process

(I) a hydrogenated ergotalkaloid or a pharmaceutically acceptable acid addition salt thereof, and

II) for the preparation of a therapeutic composition comprising low molecular weight heparin or pharmaceutically acceptable salts thereof.

As the hydrogenated ergot alkaloids (component I) of the composition, the compounds of the formula I in which

R is hydrogen or (1-4C) alkyl,

R 1 is methyl, ethyl or isopropyl,

Ra is isopropyl, secondary butyl, isobutyl or benzyl, and

X is hydrogen or methoxy, or pharmaceutically acceptable acid addition salts thereof. ~

Particularly preferred as Component I is the use of dihydroergotamine, a composition of 6nor-6-isopropyl-9,10-dihydro-2'O-methyl-5-tert-benzyl-ergopeptin and dlhydro-valine, and pharmaceutically acceptable acid addition salts thereof.

The salts of the hydrogenated ergot alkaloids are mainly methanesulfonates, maleates and tartrates. Hydroergotamine is added in particular in the form of its methanesulfonate. The "low molecular weight heparin (component II of the composition) of the present invention uses heparin obtained, for example, by isolation of the natural components of natural heparin or by cleavage of the natural heparin chain, thereby substantially reducing the average molecular weight of heparin.

The low molecular weight heparin used in the present invention has an average molecular weight of about 10,000 or less, in particular about 8,000 or less. Preferably, the average molecular weight is not less than about 4000, in particular about 5000 to about 8000, for example about 7000 ± 1000 or about 6000 ± 1000 molecular weight heparin. It is further preferred that the molecular weight distribution of component II) is relatively uniform, i.e., for example, at least 60 percent, particularly 80 percent, of the polymer units have molecular weights within the above ranges, i.e., molecular weight 10,000 or less.

The pharmaceutically acceptable salts of low molecular weight heparin used as component II are calcium and potassium salts, in particular sodium salts.

Low molecular weight heparin or its pharmaceutically acceptable salts used as component II) can be prepared by known methods, for example by separation of low molecular weight fractions derived from natural heparin, for example by fractional precipitation and ultrafiltration, as disclosed in U.S. Patent No. 2,945,591. It is disclosed either by depolymerization of high molecular weight fractions of natural heparin, for example by chemical degradation, as described, for example, in Belgian Patent 888 864 or European Patent Application 27 089. However, the heparin used as component II) can also be prepared by combinations of these methods, for example, by separating the low molecular weight constituents from the natural heparin and then depolymerizing the remaining high molecular weight constituents to give the low molecular weight heparin optionally. obtained with low molecular weight heparin.

According to the invention, it is preferable to use a low molecular weight heparin that has not undergone any - or no significant - chemical change.

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In the therapeutic composition of the invention,

Components I) and II) are present in a ratio such that 300-7000 IU (for example 500-70,000), preferably 300-35,000 IU (International Units) of Component II per 1 ml of Component I). It is particularly preferred that the components I) and U) are present in a ratio such that 1000 to 20,000 (e.g. 2000 to 20,000), especially 1000 to 10,000 IU Π0 components per 1mg of component I). If the components I) and / or Π) are present in the form of their pharmaceutically acceptable salts, an equivalent amount of the respective salt corresponding to the particular ratio of the components should be used.

The composition of the invention is conveniently administered by injection. Accordingly, the composition should be liquid. Although it would be possible to pre-mix a simple aqueous or aqueous alcoholic solution, this method is not recommended as components (1) and (Π) would react to form soluble salts in the absence of a stabilizer. Therefore, such simple solutions have no practical significance due to their low storage capacity.

Preference is given to compositions such as those disclosed in United States Patent 4,402,949, which contain a pharmaceutically acceptable calcium or magnesium salt of ethylene diamine tetraacetic acid (EDTA) as an additive (component III). In these compositions, the solvent medium preferably comprises water (component IV) and a pharmaceutically acceptable mono or poly alcohol (component V).

Among the calcium and magnesium salts of EDTA, the use of monomagnesium and monocalcium salts as component III) is recommended. Additionally, pharmaceutically acceptable multi-metal salts containing monovalent metal ions, such as sodium or potassium, in addition to calcium and magnesium ions can be used. The present invention preferably utilizes the monocalcium disodium salt (CaNa ₂ -EDTA), known as calcium titriplex. Another example is the monomagnesium dipotassium salt (MgK1-EDTA).

Component III is preferably used in an amount of from 1 to 50 mg, in particular from 1 to 25 mg, most preferably from 1 to 10 mg, relative to 5000 IU of component II). The IV) and Vj components of the solvent mixture are present in an amount of 45-72% and 28-55% of the total volume of the composition. As component V, ethanol, propylene glycol, polyethylene glycol (average molecular weight about 400), diethylene glycol, triethylene glycol, glycerol and mixtures thereof are preferred. Preferably, component V consists of a) ethanol and b) triethyleneglycol, or c) glycerol and d) propylene glycol. In such mixtures, components a) and b) are preferably present in a weight ratio of from 1: 6 to 1:10, in particular of a weight ratio of 1: 8, and components c) and d) are preferably in a weight ratio of 1: 8 to 1:12, especially in a 1:10 weight ratio.

According to a preferred embodiment of the invention, the solutions of components 1) and II) contain, as an additional component VI), other physiologically acceptable anesthetics. As anesthetic component VI), it is preferable to use an acetanflide-type active ingredient. These include, for example, acetanflides, especially those (N-phenyl) -2-aminoacetanilides and pharmaceutically acceptable salts thereof, which are anesthetic. Preferred Acetanflides are N- (2,6-Dimethylphenyl) -2- (diethylaminoacetamide) known as lidocaine, N- (2-Chloro-6-methylphenyl) -2- (butylamino) known as Hostacaine. acetamide and 2- (2-diethylaminoacetamido) -m-toluic acid methyl ester known as bayeain.

Commonly used salts of these acetanilide derivatives are, for example, the hydrochlorides. When component VI) is used, the ratio is 1-2% by weight of the composition.

It is further noted that while the solutions described above are most suitable for use in the present invention, it is possible to choose other dosage forms. Other dosage forms are exemplified by the Hophilized Formulations, which are reconstituted prior to administration. The compositions of the present invention further include additives such as carriers, diluents, stabilizers, preservatives, dyes and surfactants, and the like. contain.

The therapeutic composition of the present invention is used to prevent postoperative thrombosis.

The beneficial anti-thrombotic activity of the composition of the invention is particularly well demonstrated by the ¹²⁵ J-fibrinogen assay.

The principle of this assay is the external recording of ^ Sj.fjjjj-jnogenic radiation. E is selectively enriched in the thrombotic material of the leg veins [K. H. Frey et al., Med. 70, pages 1553-1558, in particular page 1555 (1975)].

Comparison of the composition of the invention with the composition disclosed in German Patent Publication No. 2,554,533, for example, shows that the compositions of the invention containing components I) and II) are administered once daily at a dose of 0.5 mg of dihydroergotamine methane. sulfonate and 2,500 IU of low molecular weight (mean weight 7,000 ± 1,000) was even reduced to 1,500 IU, the same effect was achieved as in the prevention of thromboses, especially postoperative thromboses. The preparation disclosed in German Patent Publication No. 533 was administered three times daily. The recommended daily dose of component I) is from 0.2 mg to about 1.5 mg, in particular 0.5 mg, the recommended daily dose of component II is from about 1000 IU to about 10 000 IU, especially about 1500 IU up to about 5000 IU. If desired, the composition of the invention is administered daily. It can be given in 2-4 smaller doses. Dosage administration is preferably carried out once daily in the form of a dosage unit. However, it is also possible to administer components I and II independently.

Preferably, unit dosage forms contain about 1500, about 2500, or about 500 IU of Component II) per 0.5 mg of Component I).

The composition of the present invention is prepared by mixing component I) with component Π) internally, however, it is also possible to dissolve component I) together with component II) in a pharmaceutically acceptable solvent. If solutions are prepared which also contain components ΙΠ), IV) and V) and optionally VT), the following steps may be taken:

1) Dissolve components I) and optionally IV) in a solvent medium containing component V),

2) Components II) and III) are dissolved in the solvent medium containing component IV),

3) Combine the solutions prepared according to 1) and 2) and

4) optionally adding additional amounts of components IV) and / or V) to the solution thus obtained. The process is conveniently carried out in an inert gas washer such as carbon dioxide, and the pH of the resulting solution is adjusted to 4,6 with a pharmaceutically acceptable acid such as methanesulfonate (when using the methanesulfonate of component I). If other salts of component I) are used, the pH of the solution should be adjusted with the appropriate acid. The compositions thus obtained may be formulated into the desired dosage form, for example, the resulting injectable solutions may be filled, after filtration, into injection vials (preferably under a carbon dioxide atmosphere).

The use of a composition of the invention comprising components I and II for the prophylactic treatment of thromboses, in particular post-operative thromboses, is also within the scope of the invention. For this purpose, a therapeutic composition containing 2500 mg, preferably 1500 IU of low molecular weight (average weight 7000 ± 1000) heparin in addition to 0.5 mg dlhydroergotamine methanesulfonate is particularly preferred.

The following examples illustrate the process of the invention.

Example 1

Preparation of a Heparin Sodium Fraction having a Molecular Weight of 6000 to 9000 1000 g of heparin sodium having an average molecular weight of about 15000 are dissolved in 6.66 liters of distilled water and the pH of the resulting solution is determined by means of a pH meter. The pH was adjusted to 2.7 with approximately 145 ml of 25% hydrochloric acid and was passed through a 520 b 1/2 (320 mm diameter) SchleicherSchüll pleated filter. The filtrate is pumped over one hour, protected from light, through a moiety filter membrane with a cut-off membrane of nominal 10,000 molecular weight under the following conditions:

Temperature: Room temperature

Filter: Pellicon Filter Cartridge (catalog no. PTGC 00001 Fa. Millipore)

Inlet side pressure: 3,2x10 'Pa

Pressure on retentate side 1.2 x 10 Pa total flow: ~ 200 mL / min

Flow of retentate: ~ 196 ml / min

The filtrate flow rate was ~ 4 ml / min.

After one hour of pumping, the filtrate is transferred to a separate vessel and collected. The filtration was discontinued after about 24 hours. Using a filtrate with a volume of about 1000-1200 ml, the pH of the heparin sodium solution

Adjust to EH, i.e. about 7, by adding about 3 ml of 30% sodium hydroxide solution. This solution is fraction 1.

The pH of the retentate is about 10 ml

It is adjusted to 3.5 with 30% sodium hydroxide solution and filtered under the conditions described above for an additional 24 hours. The filtrate thus obtained, adjusted to about 800 ml, was again adjusted to a pH of about 7 using a pH meter. This solution is fraction 2.

The retentate of this filtration was adjusted to pH 4.2 by addition of about 30 ml of 30% sodium hydroxide solution and was born under the conditions described above for an additional 24 hours. The pH of the resulting filtrate, about 500 mL, is adjusted to a starting pH of about 7 using a pH meter. This solution is fraction 3.

The three fractions are processed separately as follows:

The neutralized filtrate was mixed with 1.1 volumes of acetone to precipitate the heparin sodium in the filtrate. The oily viscous liquid was settled overnight and the supernatant solvent decanted. The residue is superimposed with about three times the amount of methanol and mixed thoroughly with a stirrer at about 1000 rpm. The heparin then appears as an off-white precipitate which can be granulated in a mortar under methanol. The resulting slurry was filtered and dried at 60 ° C in a vacuum oven at about 200 Pa. If the sodium chloride content is greater than 0.5%, a second lithium impact is required.

The fractions thus obtained are as follows

characterized by: Elemental Analysis 1. Faction 2 Faction 3 Faction 3 % C 23.7 24.9 24.4 H% 3.3 3.1 3.2 N% 2.9 2.4 2.7 0% 47.1 45.9 ' 46.8 S% 11.7 11.9 11.6 So% 123 11.8 113 Heparin activity (PTT assay, based on arteriolar III) WHO Stan-

~ 75 ± 5 IU / mg ~ 80 ± 5 IU / mg ~ 90 ± 5 IU / mg Anti-X activity ~ 150 ± 10 U / mR ~ 155 ± 10 U / mg * ~ 60 ± 10 U / mg Most common molecular weight ~ 6000 ± 1000 ~ 8000 ± 1000 ~ 9000 ± 1000 Sodium chloride content <0.5% <0.5% <0.5%

Example 2

Preparation of a stable solution for injection containing 1500 IU low molecular weight (6000 ± 1000 average molecular weight) heparin and 0.5 mg dihldroergotaml

a) Preparation of Dihydroergotamyl Solution In a liter mixer, 18.4 kg of propylene glycol and 1.84 kg of anhydrous glycerol are placed and stirred for 10 minutes with carbon dioxide. With additional stirring and carbon dioxide diet, 0.0286 kg of dihydroergotamyl methenesulfonate and 0.426 kg of lidocaine hydrochloride are dissolved in the solvent mixture over about 30 minutes.

b) Preparation of the low molecular weight solution of heparin is placed in a liter mixer with 18.4 kg of water (for injection) and stirred for 10 minutes under carbon dioxide diet. Continued stirring and carbon dioxide transfer over a period of about 30 minutes resulted in 85.71 million IU of low molecular weight (6000 ± 1000 average molecular weight) heparin (corresponding to 1.7-2.5 kg of low molecular weight heparin, according to activity) and 0.114 kg of calcinium tririplex (i.e. monocalcium disodium salt of ethylenediaminetetraacetic acid) are dissolved in water.

(c) Mixing the solutions obtained in (a) and (b)

The solution of (b) is added to the solution of (a) while passing carbon dioxide. The vessel in which solution b) was prepared is then rinsed with 1 kg of water for injection and this wash liquid is also added to the solution prepared in a). The combined solution was stirred for 10 minutes under carbon dioxide bubbling (the pH of the combined solution should be about 5.5) and then made up to a final weight of 42.810 kg (corresponding to a volume of 40 liters) by adding water for injection.

d) Filtration di) Prefiltration: Prefiltration is performed using a 0.2 gm-Ultipor pm Pali membrane filter.

d ₂ ) Sterile Filtration: Filter the solution prepared in (c) and pre-filtered using carbon dioxide directly through a sterilized pressure filtration device fitted with a membrane filter (0.2 prn-Ultipor μτη Pali) directly to the filling machine under a pressure of 1.7 bar.

The sterile-filtered solution is aseptically filled into 1 ml ampoules (0.8 ml fill volume).

Example 3

For the preparation of a solution for injection containing 0.5 mg Dihydroergotamine and 2500 IU and 5000 IU low molecular weight heparin

Following the procedure of Example 2 was repeated by using the indicated ingredients, except that the used therein heparin sodium l ^2/3 times or 3l / 3x (1000 weight average molecular weight of 6000 ±) heparin sodium replaced. This gives the title preparation.

Example 4

Preparation of a solution for injection containing 0.5 mg Dlhldroergotamine methanesulfonate and Ids (mean molecular weight 7000 ± 1000 and 8000 ± 1000) heparin.

Using the procedure of Examples 2 and 3, however, using 1) 7000 ± 1000 of average molecular weight heparin sodium as used in Examples 2 and 3 (6000 ± 1000 average molecular weight) of heparin sodium gives the title preparation. In each case, the relative amounts of the components remain the same as in Example 2. The relative amount of heparin sodium used is about 3,000, about 5,000, and about 10,000 per 1 mg of dihyd10 roergotamine methanesulfonate.

Claims (6)

PATENT CLAIMS 1) the component I) and optionally an anesthetic acetanilide derivative were dissolved in a pharmaceutically acceptable solvent containing mono''θ or polyalcohol; A process for the preparation of a therapeutic composition for thrombosis using a hydrogenated ergot alkaloid or pharmaceutically acceptable acid addition salts thereof (component 0) and heparin or a pharmaceutically acceptable salt thereof (component II), characterized in that I) the component used is dihydroergotamine and component II) heparin having a molecular weight of 10,000 or less, preferably 4000 to 10,000, in a weight ratio of 1000 IU to 10,000 IU25, and components I) and II) are intimately mixed or the two components are pharmaceutically dissolved in a harmless solvent, or 2. The process of claim 1 wherein the dihydroergotamine is used in the form of its methanesulfonate, maleinate or tartrate. 2) dissolving component II) and the magnesium or calcium salt of ethylenediaminetetraacetic acid in a solvent medium containing water and then 35 3. The process of claim 1, wherein the low molecular weight heparin is used in the range of 5000 to 8000 molecular weight heparin. 3) combining the solutions prepared according to 1) and 2), and 4. The method of claim 1, wherein the low molecular weight heparin is 7000 ± 1000 molecular weight heparin. 4) optionally adding additional water and / or mono- and / or polyalcohols to the solution thus obtained. 5. The method of claim 1 wherein je position 1, one emezve encountered particularly to small heparlnként used ^ou 6,000 ± 1,000 molecular weight heparin. 6. The process of claim 1 wherein the low molecular weight heparin is used in the form of its sodium, potassium or calcium salt.