CS257788B2 - Method of tonic pharmacopoeical preparation production - Google Patents

Abstract

for the Contracting States : BE, CH, DE, FR, GB, IT, LI, LU, NL, SE 1. A topical pharmaceutical preparation for the promotion of granulation and epithelization of wounds, containing a pharmacologically compatible carrier and an active substance, characterized by the fact that the carrier mainly consists of water or occurs in the form of a hydrated gel or as a dry gel and that the active substance increases the calcium-permeability of the plasma membrane and comprises either a) an ionizable mixture of pharmacologically compatible Ca**2+ and K**+ salts wherein, the Ca**2+ ions in the carrier occur at a concentration of 20 to 100 mM in relation to the water contained therein and the K**+-ions at a concentration of 25 to 60 mM ; or b) a Ca**2+-ionophore at a concentration of 3 x 10**-8 to 3 x 10**-6 M in combination with calcium salts at a concentration of 0.1 to 50 mM ; or c) a calcium-agonist at a concentration of 10**-5 to 10**-9 M in the presence of K**+-ions at a concentration of 5 x 10**-3 to 2 x 10**-2 M and Ca**2+ at a concentration of 10**-3 M to 3 x 10**-2 M. for the Contracting State AT 1. Process for the manufacture of a topical pharmacentical preparation for the promotion of the granulation and epithelization of wounds, containing a pharmacologically compatible carrier and an active substance, characterized by the fact that the carrier consists mainly of water or occurs in the form of a hydrated gel or as a dry gel and that the active substance increases the calcium-permeability of the plasma membrane and comprises either a) an ionizable mixture of pharmacologically compatible Ca**2+ and K**+ salts, wherein the Ca**2+ ions in the carrier occur at a concentration of 20 to 100 mM in relation to the water contained therein and the K**+ ions at a concentration of 25 to 60 mM ; or b) a Ca**2+-ionophore at a concentration of 3 x 10**-8 to 3 x 10**-6 M in combination with calcium salts at a concentration of 0.1 to 50 mM ; or c) a calcium-agonist at a concentration of 10**-5 to 10**-9 M in the presence of K**+-ions at a concentration of 5 x 10**-3 to 2 x 10**-2 M and Ca**2+ at a concentration of 10**-3 M to 3 x 10**-2 M ; and that in a pharmacologically compatible carrier, optionally in addition to other active substances and/or excipients, there is uniformly distributed an active substance a), b) or c).

Description

The present invention relates to a process for the production of a topical pharmaceutical preparation for ^: promoting wound granulation and epithelization comprising a pharmacologically compatible carrier and an active ingredient.

In vitro cell culture assays have shown that Ca ²⁺ ions have an effect on cell division. (Proc. Nat. Acad. Sel. 70, pp. 675-679). It is further known that depolarization of membrane potential can be achieved by increasing extracellular K ⁺ -ions. However, nothing is known about the effect of these effects on wound granulation and epithelization. However, extensive research has shown that even those Ca ²⁺ -ion concentrations that have been found to be optimal for growth in cell cultures, and higher Ca ²⁺ -ion concentrations do not have a supportive or inhibitory effect on wound healing.

Also, the addition of physiological amounts of potassium in the extracellular region does not promote wound healing, even though optimal extracellular concentrations of C and ²⁺ ions are also taken care of.

Surprisingly, it has now been found that Ca ²¹ · and К ^ь -iontу together at quite certain concentration ratios exhibit unexpectedly good and, at ideal molar ratios, even a very strong effect on wound granulation and epithelization, and thus in combination are a valuable means of promoting wound healing.

Concentration ratios according to the invention can be achieved when it is satisfied that the wound surface is immediately contacted with a water-containing electrolyte containing 20 to 100 mM Ca ²⁺ -ions and 25 to 60 mM K ² relative to the water contained in the electrolyte. -ions.

The electrolyte itself may consist of an early secretion and / or an aqueous preparation applied) additionally to the wound. ·

In the first case, physiologically compatible ionizable calcium and potassium salts are applied to the wound together with the solid carrier at a molar ratio of 1: 3 to 4: 1 in an amount such that the calcium and potassium salts form solutions in the above-mentioned concentration range in the early secretions. However, since the amount of secretion released varies and can only be inaccurately estimated, dry gels, dusts, ion exchangers, textiles, impregnated dressing materials, polysaccharides or similar dry compositions do not always ensure that the concentration range of the invention is achieved and maintained for a longer period of time.

Therefore, preparations which are produced in dry form and also partly in the form of such a form and which are intended to be added to them with a relatively precise amount of water before use are preferable. These are applied to the wound only after receiving this amount of water and the dissociation of Ca ²⁺ - and K ⁺ -salts. This includes, for example, dry gels according to DE-OS 28 49 570.

Mixed forms may also be contemplated where the dry carriers are first wetted with water containing Ca ²⁺ - and KMonts dissolved in the desired ratio. These include, for example, ion exchangers, gels suitable for gel chromatography (wet-meshed sieves), or simple, solid, for example wound dressings, which are moistened and finally sterile sealed.

The latter formulations are in themselves typical aqueous formulations. This is understood to mean preparations in which the carrier comprises, in particular, water containing Ca ²⁺ ions as well as K ^{+ ions} , optionally in addition to other excipients or active substances, in a concentration of 20 to 100 mM or 25 to 60 mM, respectively. A preferred range here is 25-35 mM Ca ²⁺ -ions and 35-45 mM K ⁺ -ions. A ratio of 30 mM is particularly preferred ^. Ga ²⁺ -ions and 40 mM K ⁺ -ions.

Suitable aqueous preparations are all pharmaceutical preparations which have a water content which permits the adjustment of the molar concentrations of the ions according to the invention. These include, in addition to simple aqueous solutions, lotions or oil / water emulsions, viscous solutions, dispersed systems or foams.

The above gel compositions, such as polyacrylamide / agar gels, are particularly well tolerated on wounds and are therefore preferred.

For the above preparations, the production method is advantageous, characterized in that, in addition to other auxiliary and / or active substances, based on the water contained, in addition to other auxiliary and / or active substances, up to a concentration of 20 to 100 ( mMol) Ca ²⁺ ions and up to a concentration of 25-60 (mMol) K ⁺ ions forming the pharmacologically acceptable salts and these ions are uniformly distributed in the aqueous phase.

Suitable pharmacologically acceptable salts are, in particular, chlorides and phosphates, but other inorganic or organic salts, such as citrates, malenates, succinates or the like, can also be used, if they are tolerated by tissues and are isolated. ciitelný.

In the case of aqueous preparations, it is advisable to adjust the scientific phase with isotonic agents. VéstοΊζβ'ρτο * lead, to: eliminate, foreign ions, using glur goats. Cationic effects. electrolytes, which are added in addition to Ca ²⁺ - arKMontů; no * are still explored. In many cases, it is also good to add surfactants to the aqueous phase, which increase skin permeability; for. elekt trolyty. Since the efficacy of the above formulations rests solely on the protected ion ratio of Ca ²⁺ - aK ⁺ -ions: outside the cell space, those formulations which contain said ions as the only active substance are, of course, fully effective: In many cases however, it is desirable to add other active ingredients such as antibiotics, fungistatics and superficial anesthetics.

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In the research of the mechanism described has been further found that the positive effect on the healing and granulation aid epithelisaci occurs only in the presence of Ca ²⁺ -ions K *, but whenever increases the permeability of plasma membranes and calcium ions Ca ² * - may thus flowing into the cell. This can be achieved not only as described in the foregoing by 0.2 to 10% by weight of the active compound mixture consisting of pharmacologically acceptable toniZiO2 evenly distributed in the pharmacologically acceptable carrier, optionally along with other active and / or excipients. of Ca ² * and K * salts in a Ca ² *: K ⁺ molar ratio of from 1: 3 to 4: 1, thus using K * ions to open the so-called calcium channels in the plasma membrane, but also by mixtures of the active ingredients in a pharmacologically acceptable carrier finely disperse either Ca ² * -iontophor at a concentration of 3X10 ^-8 to 3X10 ⁶ M in combination with calcium salts at a concentration of 0.1-50 mM, or a calcium agonist at a concentration of 10 ⁵ to 10 ^-9 M in the presence of K @ ⁺ -Of.sub.2 at a concentration of ^{5 *} 10 @ ³ to 2 * 10 @ ² M and Ca @ + at a concentration of 10 @ ^-3 M to ³ * 10 @ ^-2 M and the preparation obtained is used as a wound healing agent.

Ga ^2'- iontophores are compounds that selectively permeabilize the plasma membrane for calcium.

Such an ion is, for example, an antibiotic known first under the designation A 23 187, i.e. 6S [6alpha (2S *, 3S ^X ), 8beita (R * J, 9beta, Haifa) -5-methylamino-2- [3,9] , 11-trimethyl-8- [1-Imethyl-2-oxo-2- (1H-pyrrol-2-yl) -ethyl] -1,7-dioxa-spiro (5,5-undec-2-yl) methyl] - 4-Beoxazolecarboxylic acid (Hoechst Doc. No. 8154-1082).

In combination with calcium 23187 A operates according to the invention at a concentration of 3X10 ⁸ to 3X10 ~ ⁱ n, wherein the calcium salt in the extracellular space may к available only in a concentration of 0.1 to 50 mm to ensure a sufficient intake of Ca ²⁺ -ions * do bu'пёк.

Since this is principally a calcium deposition of cells through the plasma membrane, the effect of faster wound healing according to the invention can be achieved by all other Ca-ionophores.

Similarly, Ca-agonists also apply because they also increase the plasma membrane permeability to calcium. They open calcium channels in the presence of potassium. When using the Ca-agonists according to the invention, only very little potassium is required, so that, especially in wounds with a large area of potassium management, it is not disturbed. This represents an improvement over the use of pure Ca ¹ * - and K * -ions as described above.

The most effective Ca-agonist concentration range is 10 ^-5 to 10 ^-9 M. The required potassium concentration is in the range of 5X10 ^-3 to

2X10 ' ² M. The calcium concentration should be between ³ ” ³ M to 3X10 ~ ² M.

Such a calotte agonist is, for example, Вау К 8644, which is known to stimulate the calcium supply to the cell [cf., for example, Arzneimittelforschung Drg. Res. 33 (113), No. 9 (1933), and Biochem. and Biophys. Research Communic. (1984) 118, No. 3, pp. 842-847].

It is also extremely surprising that inside the cell calcium is an essential factor in wound granulation. The concentration of calcium in the extracellular space without permeability enhancers according to the invention has no effect on wound healing. Neither the removal of calcium by the addition of a calcium chelating agent such as ethyleneglyc-ol-bis (β-aminoethyl) -N ^ XX-tetraacetic acid (EGTA) nor an increase in calcium ions by 20 times the physiological concentration in the cellular space leads to a change in granulation of the wounds . From this, one skilled in the art could only conclude that calcium ions do not participate significantly in the granulation of wounds.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for promoting wound granulation and epithelization, comprising a pharmacologically compatible carrier and an active ingredient, comprising the use of an active substance that increases the plasma membrane permeability to calcium and consists of either

a) based on the carrier, 0.2 to 10% by weight of an ionizable mixture of pharmacologically compatible salts of Ca. ² * and K * in a Ca ² *: K * molar ratio of 1: 3 to 4: 1, or

b) ^Ca2 * concentration iontóforu 3X10 ^-8 to 3X10 ^-6 M in combination with calcium salts at a concentration of 0.1 to 50 mm, or

c) agonists, calcium at a concentration of 10 @ ⁵ to 10 ⁹ in the presence of ions K + in a concentration of 5X10 ³ to 2X10 ^'2 M and Ca ² * at a concentration of 10 ^-3 M to 3X10 ^-2 M, and the carrier consists essentially of water and optionally, in addition to other excipients and / or active ingredients, based on the water contained, up to a concentration of 20 to 100 mM Ca 2+ and 25 to 60 mM K ² ions, and these ions are uniformly dispersed in the aqueous pharmaceutical base.

In a preferred embodiment of the process of the invention, the carrier comprises a hydrated gel containing methylhydroxycellulose.

The following drawings illustrate the invention in more detail:

Giant. 1 depicts the dependence of granulation tissue formation on extracellular calcium concentration. In this figure, the ordinate shows the amount of granulation tissue formed in mg per wound. Ca ² * (molar concentration) concentrations are applied to the abscissa axis, quenching by a factor of 10.

2 $ 7788

Giant. 2 shows the dependence of granulation tissue formation on the concentration of calcium permeability enhancers. In this figure, the ordinate shows the amount of granulation tissue formed in mg per wound. On the abscissa axis, molar concentrations of substance A 23 187 are reported, alternatively with (3 mM) EGTA or ¹ (1 mM) Ca ²⁴ , optionally with or without this addition (number 5).

Figure 1 shows that the formation of granulation tissue is essentially independent of the extracellular calcium concentration.

FIG. 2 shows that the active ingredient increasing the permeability, i.e. A 23187 in the presence of calcium ions in the extracellular space clearly increases the granulation of wounds at a concentration between 10 ^-7 to 3X10 ^-6 M, especially in a concentration of 3X10 ^-6 M over comparative Sample 5. When the Ca ²⁺ concentration in the cell decreases by complexation with EGTA in the extracellular space and is used as an A 23 187 permeability enhancer, the granulation clearly decreases. This experiment demonstrates that the formation of granulation tissue, a presumption of wound healing, is significantly dependent on the Ca ²⁺ content of the cells and that granulation can be significantly increased when the Ca ²¹ content of the cells is increased by the permeability enhancers of the invention. The experiments underlying the figures for Figures 1 and 2 were performed as follows:

The skin on the backs of the guinea pigs was cut through to the fascia. Teflon ring diameter 21 mm was sewn into the wound. This prevented the epithelial wound closure. A polyacrylamide / agar gel containing the active ingredients at the indicated concentration was applied to the dorsal muscle fascia, which was free of granulation tissue at the time of surgery. After a period of three days, when the optimum granulation was reached, all the granulation tissue was collected with a sharp spoon, weighed and processed histologically. In method a) carried out at a concentration of 30 mM Ca ²⁺ -ions and 40 mM K + -ions, with 0.9 wt% NaCl added to the gel for isotonicity, the granulation tissue amount increased significantly by up to about 180% . Histological controls have shown true cell multiplication, not just an increase in cell volume.

The following examples serve to further illustrate the invention.

Example 1

Isotonic solution

Place 80 ml of purified water in a 100 ml collection flask. While stirring with a magnetic stirrer, 20 mg of benzalkonium chloride is added and dissolved. Then 0.3 g of potassium chloride, 0.44 g of calcium chloride (both Ph. Eur. I. compliant salts) and 2.62 g of glucose monohydrate (Ph. Eur. II) are added sequentially. The flask is allowed to warm to a temperature of 20 ° C in a water-bath and then made up to 20 ° C with warm purified water. The solution is then sterile filtered through a 0.2 μκη porosity membrane filter and sterile filled.

(0.29822 g KCl 40 mMol K ⁺ , 0.44106 g CaCl 2 30 mMol Ca ²⁺ ).

Example 2

Oil in water emulsion

In the first portion, 7 g of a mixture consisting of saturated fatty acids, fatty alcohols, lanolin, mineral oils and non-ionic emulsifiers together with 2.5 g of fatty acid ester, polyethylene glycol and glycerin are melted homogeneously by heating to 70 ° C in a water bath. g of stearic and palmitic acid monoglycerides, 0.3 g of cetyl alcohol and 3.0 g of isopropyl palmitate.

In a second crop, 80 g of purified water are mixed with 3 g of propylene glycol while stirring and heated to 70 ° C. 0.3 g of potassium chloride, 0.44 g of calcium chloride and 0.2 g of preservative are then added to the mixture. The clear solution obtained is emulsified into the first crop with stirring at 70 ° C. The emulsion thus obtained is cooled to 40 ° C and the water loss due to evaporation is compensated. The emulsion cooled to 30 ° C is then filled.

Example 3

Transparent liquid dressing material (gel bowl)

Handle A:

3.5 g of acrylamide and 0.091 g of bisacrylamide are dissolved in 100 purified water. The solution is then heated to 60 ° C.

Handle B:

Dissolve 0.3 g of potassium chloride and 0,44 g of calcium chloride in 100 ml of purified water and add 0,2 g of preservative. After addition of 2 g of agar (OAB9), the solution is heated to boiling under magnetic stirring and then cooled to 60 ° C.

The batches A and V are then mixed at 60 ° C with stirring. 0.045 grams of ammonium persulfate and 0.045 g (60 .mu.l) of tetramethylenediamine are then added. Brief vigorous stirring is followed by pouring onto petri dishes that have been preheated to 60 ° C in an oven. The filled Petri dishes are then stored for 30 minutes at 56 ° C in an oven. It is then allowed to cool to room temperature and the dishes obtained with transparent solidified plates are placed in a thermostat at 4 ° C for 24 hours to mature. The gel dishes thus obtained can be used immediately to cover the wounds.

Example 4

The dispersion gel g of purified water is heated to 70 ° C and 0.3 g of potassium chloride and 0.44 g of calcium chloride are added. After addition of 0.2 g of preservative, 5 g of methylhydroxyethylcellulose is dispersed in the prepared solution. It is then cooled with stirring. Upon cooling, a highly viscous, air-bubble gel having a viscosity of 90 Pa is obtained. s, which can be used immediately to treat wounds.

Example 5

Textile wound cover

Sterile 4X4 cm and 5 mm thick mule shreds are immersed in the sterile isotonic solution of Example 1 and then squeezed only to the extent that they are no longer cut from the shred. The pieces thus obtained are then sealed under sterile conditions into a polyethylene film.

Example 6

Rubbing gel with fabric A 23 187

000 g of purified water are heated to 70 ° C and 1.6 mg of substance A 23 187 and 560 mg of calcium chloride are added. After addition of 2 g of preservative, 50 g of methylhydroxyethylcellulose are dispersed in the solution obtained. It is then cooled with stirring. Upon cooling, a highly viscous, air-bubble gel having a viscosity of 90 Pa is obtained. s, which can be used immediately to treat wounds.

Example 7

Spreadable gel with Вау К 8644 fabric

000 g of purified water are heated to 70 ° C and 1.49 g of potassium chloride, 3.3 g of calcium chloride and 3.56 mg of Вау К 8644 are added. After adding 2 g of preservative, it is dispersed in the obtained solution of 50 mg. methylhydroxyethylcellulose and processed as in Example 1.

Claims (6)

SUBJECT OF THE INVENTION A process for the preparation of a topical pharmaceutical preparation for the promotion of wound granulation and epithelization, comprising a pharmacologically compatible carrier and an active substance, characterized in that an active substance is used which increases the plasma membrane permeability to calcium and consists either of (a) on a carrier, 0,2 to 10% by weight of the ionised mixture of pharmacologically compatible salts of Ca ² H and K ⁺ in a molar ratio of Ca ²⁺ : K ⁺ 1: 3 to 4: 1, or b) on the Ca ²⁺ concentration iontoforu 3X10 ^-8 to 3X10 ^-6 M in combination with calcium salts at a concentration of 0.1 to 50 mm, or c) agonists, calcium at a concentration of 10 ⁵ to 10 ^"9 in the presence of K in a concentration of 5X10 ⁴ to 2X10 ^{-3 '2} M, and Ca at a concentration of ^{24 -3} M ΙΟ to 3 x 10 ^-2 M, and the carrier essentially consists of % of water and optionally added in addition to other excipients and / or active substances, based on the water contained, up to a concentration of 20 to 100 mM Ca ²⁺ and 25 to 60 mM K ^+, and these ions are uniformly dispersed in the aqueous pharmaceutical base. 2. A process according to claim 1, wherein the calcium permeability enhancing agent is used and consists either of: a) based on the carrier, 0.2 to 10% by weight of an ionized mixture of pharmacologically compatible salts of Ca ^{2 h} and K ⁺ in a molar ratio of Ca ²⁺ : K ⁺ 1: 3 to 4: 1, or (b) Ca ²⁺ iontophor at a concentration of 3X10 ⁸ to 3X10 ⁶ M in combination with calcium salts at a concentration of 0,1 to 50 mM; or c) agonists, calcium at a concentration of 10 ^"5 to 10 ⁹ in the presence of K ⁺ ions in a concentration of 5X10-3 to 2X10-2 M and Ca ²⁺ kincentraci м 10 ³ to 3X10 ^'2 M, and the carrier comprises a hydrated gel containing methylhydroxyethylcellulose and, if appropriate, in addition to other excipients and / or active ingredients, based on the water contained, up to a concentration of 20 to 100 mM Ca ²⁴ and 25 to 60 mM K ¹ and these ions are uniformly dispersed in the aqueous pharmaceutical base. 3. The method according to claim 1, characterized Lim that uses active substances increasing the permeability of the plasma membrane calcium and comprising, based on the carrier of 0.2 to 10 ° / o by weight of a mixture of ionizable farmiakologicky acceptable salts of Ca + ² and K ⁴ * in a molar ratio of Ca ²⁺ : K ⁺ 1: 3 to 4: 1a, the carrier consists essentially of water and is added, if appropriate in addition to other excipients and / or active substances, based on the water contained, up to a concentration of 20 to 100 mM Ca ions ²⁺ and 25 to 60 mM K ⁺ ions, and these ions are uniformly dispersed in the aqueous pharmaceutical base. 4. A process according to claim 3, characterized in that an active substance enhancing agent is used II the plasma membrane permeability of calcium and consisting, based on the carrier, of 0.2 to 10% by weight of an ionizable mixture of pharmacologically compatible salts of Ca ²⁺ and K ⁺ in a molar ratio of Ca ²⁺ : K ⁺ 1: 3 to 4: 1a from a hydrated gel containing methylhydroxyethylcellulose and, if appropriate, added, in addition to other excipients and / or active ingredients, based on the water content, up to a concentration of 20 to 100 mM Ca ²⁺ and 25 to 60 mM K ⁺ ions pharmaceutical base. 5. The method of claim 1, wherein the calcium membrane permeability enhancer is either a Ca ²⁺ iontophor at a concentration of 3X10 ⁸ to 3X10 ⁶ M in combination with calcium salts at a concentration of 0.1 to 50. mM or an agonist at a concentration of 10 ⁵ to 10 ⁹ in the presence of K ⁺ ions at a concentration of 5X10 ³ to 2X10 ² M and Ca ²⁺ at a concentration of 10 ³ M to 3X10 ² M, and the carrier consists essentially of water and in addition to other excipients and / or active ingredients, based on the water contained, up to a concentration of 20 to 100 mM Ca ²⁺ ions and 25 to 60 mM K + ions, and these ions are uniformly dispersed in the aqueous pharmaceutical base. 6. The method of claim 5, wherein the calcium membrane permeability enhancing agent is used and consists of either a Ca2 ⁺ iontophor at a concentration of 3 * ¹⁰ < ⁸ > to 3 * ¹⁰ < ⁶ > 50 mM, or a calcium agonist at a concentration of 10 ⁵ to 10 ⁹ in the presence of K ⁺ ions at a concentration of 5X10 ³ to 2X10 ² M and Ca ²⁺ at a concentration of 10 ³ M to 3X10 ² M, and the carrier consists of a hydrated gel containing methylhydroxyethylcellulose. and optionally, in addition to other excipients and / or active ingredients, based on the water contained, up to a concentration of 20 to 100 mM Ca ²⁺ ions and 25 to 60 mM K + ions, and these ions are uniformly dispersed in the aqueous pharmaceutical base.