DD232819A5 - PHARMACEUTICAL TOPICAL PREPARATIONS, METHOD FOR THE PRODUCTION THEREOF AND THEIR USE TO PROMOTE WOUND GRANULATION AND EPITHELISATION - Google Patents

Abstract

Pharmaceutical topical preparations are described which contain a pharmacologically acceptable carrier and an active ingredient and which are characterized in that the active ingredient increases the calcium permeability of the plasma membrane and either ausa) based on the carrier 0.2 to 10 weight percent of an ionizable mixture of pharmacologically acceptable Ca2 and K salts in Ca 2: K molar ratio of 1: 3 to 4: 1 or b) of a Ca 2+ ionophore in a concentration of 310 8 to 310 6 M in combination with calcium salts in a concentration of 0.1-50 mM. orc) consists of a calcium agonist in a concentration of 10 5 to 10 9 in the presence of K ions in a concentration of 510 3 to 210 2 M and Ca in a concentration of 10 3 M to 310 2 M. The new preparations surprisingly promote wound granulation and epithelization. Furthermore, processes for the preparation of the preparations according to the invention are described.

Description

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Field of application of the invention

From in vitro studies on cell cultures, it is known that Ca ²⁺ ions exert an influence on the cell division. (Proc. Nat. Acad., 70, 675-679). Furthermore, it is known that the depolarization of the membrane potential can be achieved by increasing the extracellular K ⁺ concentration. However, nothing is known about the effects of these effects on wound granulation and epithelization. Extensive investigations have shown, however, that neither the cell concentrations optimally found for growth of Ca ²⁺ concentrations nor higher Ca ²⁺ concentrations have a promoting or inhibiting effect on wound healing.

The addition of physiological amounts of potassium in the extracellular area also does not promote wound healing, even while providing optimal extracellular Ca ²⁺ concentration.

Object of the invention

It is an object of the invention to improve wound healing.

Essence of the invention

It has now surprisingly been found that Ca ²⁺ and K ⁺ ions together in very specific concentration ratios exert an unexpectedly good and even at ideal molar ratios a very strong effect on wound granulation and epithelization and therefore in combination a valuable means of promoting the Constitute wound healing.

The concentration ratios of the invention can be achieved if it is ensured that the wound surface is directly associated with a water-containing electrolyte, based on the water contained in this electrolyte 20 to 10OmM Ca ²⁺ ions and 25 to 6OmM K ⁺ ions contains.

The electrolyte itself may consist of wound exudate and / or an aqueous preparation additionally applied to the wound.

In the former case, physiologically acceptable ionizable Ca and K salts together with a solid carrier in a molar ratio of 1: 3 to 4: 1 are applied to the wound in an amount such that the Ca and K salts in wound secretion solutions are as indicated above Make concentration range. However, since the amount of wound secretion varies and can only be estimated inaccurately, dry gels, powders, ion exchangers, nonwovens, impregnated dressings, polysaccharides or similar dry preparations do not always ensure that the concentration range of the invention can be achieved and maintained for a longer time ,

More favorable are therefore those preparations which, although produced in dry form and partly also come in this form on the market, and which are intended to be added prior to use with a relatively precise amount of water. These are only neih recording this Wasrtrmenge and took place. L isoriation of Ca ²⁺ - and K ⁺ salts brought to the wound. These include z. B. dry gels according to DE-OS 2849570.

Mixed forms in which dry carriers are moistened from the outset with water containing Ca ²⁺ and K ⁺ dissolved in the desired ratio are conceivable. These forms include, for example, ion exchangers, gel suitable for gel chromatography (molecular sieves) or simple solid, z. As textile wound dressings that are moistened and then sealed sterile.

The latter preparations are in themselves already typical aqueous preparations. This is understood to mean preparations in which, above all, water acts as a carrier and optionally contains Ca ⁺ ions and K ⁺ ions in addition to other auxiliaries and / or active substances in a concentration of 20 to 10 μm or 25 to 60 mM. The preferred range is 25 to 35 mM Ca ²⁺ ions and 35 to 45 mM K ⁺ ions. Most preferably, the ratio of 3OmM Ca ²⁺ ions and 4OmM K ⁺ ions.

Suitable aqueous preparations are all pharmaceutical preparations which have a water content which allows the adjustment of the molar ion concentrations according to the invention. In addition to simple aqueous solutions, lotions or oil-in-water emulsions, this also includes viscous solutions, dispersed systems or foams.

The above-mentioned gelatinous preparations, such as. B. polyacrylamide / agar gels are particularly wound compatible and therefore preferred.

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For the abovementioned preparations, preference is given to a process for the preparation which comprises adding up to a concentration to an aqueous pharmaceutical carrier as active ingredient in a manner known per se, if appropriate in addition to other excipients and / or active substances, based on the water contained from 20 to 100 (mmol) of Ca ²⁺ ions and 25 to 60 (mmol) of IC ions, Ca ²⁺ - and K ⁺ -ions forming pharmacologically acceptable salts and these ions evenly distributed in the aqueous phase.

As pharmacologically acceptable salts are especially chlorides and phosphates in question, but it can also be other inorganic or organic salts, such as. As citrates, maleates, succinates or similar salts can be used, provided that they are tissue-compatible and dissociable.

In the case of aqueous preparations, it is favorable to adjust the aqueous phase isotonic. This can be done preferably by means of glucose to avoid foreign ions. The effects of cationic electrolytes added alongside Ca ²⁺ and K ⁺ have not yet been explored. In many cases, it is also beneficial to add to the aqueous phase surfactants that increase skin permeability to electrolytes.

Since the effectiveness of the above-mentioned preparations is based solely on the claimed ionic ratio of Ca ²⁺ and K ⁺ ions in the extracellular space, it goes without saying that those preparations which contain the stated ions as the sole active ingredient are also fully effective. In some cases, however, the addition of other active ingredients is desirable such. As antibiotic or fungistatic agents or surface anesthetics.

In the studies on the mechanism of the described preparations, it was further found that the positive effect on the promotion of wound granulation and epithelization not only in the presence of Ca ⁺ * - and K ⁺ ions, but always occurs when the calcium permeability of the plasma membrane increased and thus Ca ²⁺ ions can flow into the cell. However, this can not only be achieved, as described above, by optionally adding, apart from other active ingredients and / or excipients, from 0.2 to 10% by weight of an active substance mixture consisting of pharmacologically acceptable ionizable Ca ²⁺ in a pharmacologically acceptable carrier and K ⁺ salts in the Ca ²⁺ : K ⁺ molar ratio of 1: 3 to 4: 1 evenly distributed, and thus K ⁺ ions used to open the so-called calcium channels in the plasma membrane, but also by the fact that instead of the described active ingredient mixture either a Ca ²⁺ -lonophor in a concentration of 3 x 10 ^-8 to 3 χ 10 ~ ^{β Μ} in combination with calcium salts at a concentration of 0,1-5OmM, or a calcium-agonist at a concentration of 10 ~ ^s χ to 10 ^-9 M in the presence of K ⁺ ions in a concentration of 5 10 ^-3 to 2 χ 1O ^-2 M and Ca ⁺⁺ at a concentration of 1O ^-3 M to 3 x 10 ^"2 M evenly distributed in a pharmacologically acceptable carrier and applying the obtained preparation as a wound healing agent.

Ca ²⁺ -lonophores are compounds that selectively permeabilize the plasma membrane for calcium.

Such an ionophore is, for example, the antibiotic 6S [6α (2S *, 3S *), 8β (R ·), 9β, 11a] -5-methylamino-243,9,11, first known under the name A23187. trimethyl-841-methyl - 2 - oxo-2- <1H-pyrrolo ^ 2- ^ l) -ethylb1,7-dioxaspirot5,5] undec-2-yl] methyll-4-benzoxazoi-carboxylic acid. (Hoechst Doc. No. 8154-1082] In combination with calcium ionophore A23187 acts according to the invention in a concentration of 3 χ 10 ^-8 to 3 χ 10 ^{"β Μ} wherein the calcium salts be present in the extracellular space only in a concentration of 0.1 to 5OmM to ensure sufficient Ca ²⁺ influx into the cell.

Since, in principle, it is the calcium supply of the cell through the plasma membrane, the effect according to the invention of faster wound healing can likewise be achieved by all other calcium ionophores.

Similarly, Ca agonists act as they also increase the calcium permeability of the plasma membrane. They open the calcium channels in the presence of potassium. In the use of Ca agonists according to the invention, however, only very little potassium needs to be applied, so that, especially in large wounds, the potassium balance is not disturbed.

This is an improvement over the use of the pure Ca ⁺⁺ and IC ions as described above.

The effective concentration range of the calcium agonist is 10 ^-5 to 10 ^-9 M. The required potassium concentration is in the range of 5 x 10 ^-3 to 2 χ 10 ^"2 M. The calcium concentration should be between 10" ³ M and 3 x 10 " ² sts.

One such calcium agonist is, for example, Bay K8644, which is already known to stimulate calcium influx into the cell. e.g. Drug discovery / drug. Res. 33 (II) No. 9 (1983) and Biochem. and Biophys Research Communic. (1984) 118, no. 3, pp. 842-847].

It is extremely surprising to note that within the cell, calcium is an important factor in wound granulation.

The calcium concentration in the extracellular space has no effect on wound healing without the permeability enhancement agents according to the invention. Neither withdrawal of calcium by adding a calcium chelator such as ethylene glycol bis (β-aminoethyl) -N, N, N ', N'-tetraacetic acid (EGTA) nor an increase in calcium concentration by twenty times the physiological concentration in the extracellular space to a change in wound granulation. From this the expert could only conclude that calcium ions are not significantly involved in wound granulation.

The present invention is therefore a pharmaceutical topical preparation for the promotion of wound granulation and epithelization, containing a pharmacologically acceptable carrier and an active agent, which is characterized in that the active ingredient the ^{Calciumpermeah: i} ity of the plasma membrane is increased and either

a) based on the carrier 0.2 to lu wt .-% of an ionizable mixture of pharmacologically acceptable Ca ²⁺ - and K ⁺ -SaI ^ e in Ca ²⁺ : K ⁺ Mo ratio of 1: 3 to 4: 1 or

b) from a Ca ²⁺ -lonophor in a concentration of 3 x 10 ^-8 to 3 x 10 ^"6 M in combination with calcium salts at a concentration of 0,1-5OmM, or

c) a calcium-agonist at a concentration of 10 ~ ^B to 10 ^"9 M in the presence of K ⁺ ions in a concentration of 5 χ 10" ³ to 2 x 10 ^"2 M and Ca ⁺⁺ at a concentration of 10 " ³ sts to 3 x 10" ² sts

consists.

Another object of the invention is a process for the preparation of a topical pharmaceutical preparation for promoting wound granulation and epithelization, which is characterized in that in a pharmacologically acceptable carrier optionally in addition to other active ingredients and / or auxiliaries an active ingredient a), b) or c) evenly distributed, as well as the use of the designated pharmaceutical preparation for the promotion of wound granulation and epithelization.

The figures are intended to explain the subject matter of the invention in more detail:

From Figure 1 it can be seen that the formation of granulation tissue is essentially independent of the extracellular calcium concentration.

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From Figure 2 shows that the permeability-increasing agent A23187 in the presence of Ca ²⁺ ions in the extracellular space, the wound granulation at a concentration between 10 ~ ⁷ and 3 χ 10 ~ ^β Μ, especially at the concentration 3 χ 10 ~ ^β Μ compared to Comparative sample 5 significantly increased. Decreasing the Ca ²⁺ concentration in the cell by Ca ²⁺ withdrawal by means of EGTA in the extracellular space and application of A23187 as a permeability-increasing agent, the granulation decreases significantly. This experiment shows that the formation of granulation tissue, a prerequisite for wound healing, is strongly dependent on the Ca ²⁺ content of the cells and that the granulation can be increased considerably when the Ca ²⁺ content of the cell is increased with the permeability-increasing agents according to the invention elevated. The experiments underlying FIGS. 1 and 2 were carried out as follows:

The dorsal skin of guinea pigs was severed except for the fascia. A Teflon ring with a diameter of 21mm was sewn into the wound. This should prevent epithelial wound closure. On the fascia of the back muscles, which was free of granulation tissue at the time of surgery, was applied for 3 days, an aqueous polyacrylamide agar gel containing the active ingredients in the indicated concentration. After 3 days, when the optimum granulation was reached, the whole granulation tissue was removed with a sharp spoon, weighed and processed histologically. In the method a) carried out with a concentration of 3OmM Ca ²⁺ ions and 40 mM K ⁺ ions, wherein the gel was added for reasons of isotonia with 0.9 wt .-% NaCl, there was a significant increase in the amount of Granulation tissue up to about 180%. The histological controls gave a true cell proliferation, not just a volume increase of the cells

 The following examples serve to explain the invention in more detail:

example 1 Isotonic solution

80 ml of purified water are placed in a 100 ml volumetric flask. 20 mg of benzalkonium chloride are then added with stirring with a magnetic stirrer and dissolved. Subsequently, 0.3 g of potassium chloride, 0.44 g of calcium chloride (both salts according to Ph. Eur. I.) and 2.62 g of glucose monohydrate (Ph. Eur. II) are added successively. The flask is heated in a water bath to 20 ⁰ C and finally filled with purified also 20 ⁰ C warm water to the mark. The solution is then sterile filtered through a membrane filter of 0.2 цт pore size and filled sterile. [0.29822 g KCl ~ 4OmMoI K ⁺ 0.44106 g CaCl ₂ ~ 3OmMoI Ca ²⁺ ]

Example 2 Oil-in-water emulsion

In a first batch, 7 g of a mixture consisting of saturated fatty acids, fatty alcohols, wool wax, mineral oils and nonionic emulsifiers together with 2.5 g of polyethylene glycol-glycerol fatty acid ester, 3 g monoglycerides of stearic and palmitic acid, 0.3 g of cetyl alcohol and 3.0 g of isopropyl palmitate by heating homogeneously melted to 70 ⁰ C in a water bath.

In a second batch, 80 g of purified water are mixed with stirring with 3 g of propylene glycol and heated to 7O ⁰ C. The resulting mixture is then added with 0.3 g of potassium chloride, 0.44 g of calcium chloride and 0.2 g of a preservative. The resulting clear solution is emulsified with stirring at 70 ⁰ C in the first batch. The emulsion thus obtained is cooled to 40 ⁰ C and the loss of water suffered by evaporation supplemented. The cooled to 30 ⁰ C emulsion is then filled.

Example 3 Transparent fluid dressing material (gel plate)

Approach A:

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

Approach B:

0.3 g of potassium chloride and 0.44 g of calcium chloride are dissolved in 100 ml of purified water and mixed with 0.2 g of preservative. After adding 2 g of agar (ÖAB9), the solution is brought to the boil with stirring with a magnetic stirrer and then cooled to 60 ⁰ C.

Subsequently, the mixtures A and B are mixed at 60 ^° C. with stirring. Thereafter, 0.045 g of ammonium peroxydisulfite and 0.045 g (60 Kg) of tetramethylenediamine are added. After brief vigorous stirring, the pouring into Petri dishes, which are previously preheated in the oven to 60 ⁰ C. The filled Petri dishes are then stored for 30 minutes in a set at 56 ° C oven. The mixture is then allowed to cool to room temperature and the shells with the obtained transparent solidified plates for 24 hours in a tempered at 4 ° C cabinet for maturation. The plates thus obtained can be used directly to cover wounds.

Example 4 Spreadable gel

94 g of purified water are heated to 70 ⁰ C and treated with 0.3 g of potassium chloride and 0.44 g of calcium chloride. After addition of 0.2 g of preservative 5 g of methyl hydroxyethyl cellulose are dispersed in the resulting solution. Then it is cooled with stirring. After cooling, a highly viscous aerated gel with a viscosity of 90 Pas is obtained, which can be used directly for wound treatment.

Example 5 Textile wound dressing

4 x 4 cm and 5 mm thick sterile gauze lobules are immersed in the sterile isotonic solution according to Example 1 and then only squeezed so far that the lobules no longer drip. The lobules thus obtained are then sealed under sterile conditions in polyethylene film

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Example 6 Spreadable gel with A 23187

1000 g of purified water are heated to 70 ⁰ C and treated with 1.6 mg A23187 and 560 mg calcium chloride. After addition of 2 g of preservative 50 g of methyl hydroxyethyl cellulose are dispersed in the resulting solution. Then it is cooled with stirring. After cooling, a highly viscous aerated gel with a viscosity of 90 Pas is obtained, which can be used directly for wound treatment.

Example 7 Spreadable gel with Bay K 8644

1000 g of purified water are heated to 70 ⁰ C and treated with 1.49 g KCI, 3.3 g CaCl ₂ and 3.56 mg Bay K 8644 added. After addition of 2 g of preservative 50 g of methyl hydroxyethyl cellulose are dispersed in the resulting solution and worked up as in Example 1.

Claims (6)

-1- 760 46 claims: 1. A process for the preparation of a pharmaceutical topical preparation for promoting wound granulation and epithelization, comprising a pharmacologically acceptable carrier and an active ingredient, characterized in that the active ingredient increases the calcium permeability of the plasma membrane and either from a) based on the carrier 0.2 to 10 wt .-% of an ionizable mixture of pharmacologically acceptable Ca ²⁺ and K ⁺ salts in Ca ²⁺ : K ⁺ molar ratio of 1: 3 to 4: 1 or b) from a Ca ²⁺ -lonophor in a concentration of 3 x 10 ^-8 to 3 χ 10 ^"6 M in combination with calcium salts at a concentration of 0,1-5OmM, or c) from a calcium agonist in a concentration of 10 ~ ^s to 10 ~ ⁹ in the presence of K ⁺ ions in a concentration of 5 × 10 ^-3 to 2 × 10 ^-2 M and Ca ⁺⁺ in a concentration of 10 " ³ sts to 3 x 10" ² sts consists. 2. A process for the preparation of a pharmaceutical preparation according to item 1, characterized in that the carrier consists predominantly of water. 3. A process for the preparation of a pharmaceutical preparation according to item 1 and 2, characterized in that the carrier is in the form of a hydrous gel. 4. A process for the preparation of a pharmaceutical preparation according to item 1, characterized in that a dry gel is used as the carrier. 5. A process for the preparation of a pharmaceutical preparation according to item 1 a), characterized in that based on the water contained 20 to 10OmM Ca ²⁺ ions and 25 to 60 mM K ⁺ ions. 6. A process for the preparation of a pharmaceutical preparation according to item 5, characterized in that it contains about 3OmM Ca ²⁺ ions and about 4OmM K ⁺ ions.