SK15212000A3 - A preparation consisting of calcium hydroxide, a dihydric or polyvalent alcohol, a fatty oil of vegetable or animal origin, and its use for regenerating collagen - Google Patents

Abstract

The invention relates to a preparation containing calcium hydroxide, a dihydric or polyvalent alcohol, a fatty oil of vegetable or animal origin and possibly pharmaceutically compatible adjuvants. The invention also relates to the production of such a preparation, the use of such a preparation for the regeneration of collagen and the use of such a preparation for the production of a medicine which promotes the in vivo regeneration of collagen.

Description

A preparation consisting of calcium hydroxide, a dihydric or polyhydric alcohol and a fixed oil of vegetable or animal origin and its use for the recovery of collagen

Technical field

The invention relates to a composition composed of a fixed oil of vegetable or animal origin, calcium hydroxide and a dihydric or polyhydric alcohol, and suitable pharmaceutically acceptable excipients for the preparation of such a composition, use of such a composition for collagen regeneration and use of such composition for producing a drug for inducing collagen regeneration in vivo .

BACKGROUND OF THE INVENTION

Bone consists of about 60% mineral substance (hydroxyapatite, calcium phosphate) and about 40% organic matter, mainly collagen. Bone metabolism is predominantly determined by the interplay of bone forming cells (osteoblasts) and bone degrading cells (osteoclasts and osteocytes), whose action is in a balanced bone in healthy bone.

Bone formation can be divided into two main phases (a) organic tissue synthesis (collagen synthesis) and (b) subsequent incorporation of the mineral substance into a preformed organic matrix mediated by so-called matrix vesicles.

The connective protein tissue of collagen represents the majority of organic substances in the bone. The protein consists of three helically twisted polypeptide chains in which the amino acid composition can vary, resulting in a variety of different types of collagen. It is common to all types of collagen that collagen fibers have exceptionally high mechanical strength. This strength is based on a multiplicity of intramolecular and intermolecular bonds of collagen fibers, which in this way from a dense network of collagen fibers VY2> ^μ. '

- t face connective tissue. As already mentioned, bone tissue is formed by incorporating mineral substances (hydroxyapatite and calcium phosphate) into this network. Bone building as a result of growth and regeneration processes is always preceded by collagen biosynthesis.

Until now, in any case of bone damage, the bone regeneration process has been left to itself, at most supported by antibiotics and corticoids to avoid any risk of infection endangering the healing process.

Some factors capable of influencing bone formation and regeneration have also been described. These are mainly physical factors (mechanical and electrical forces), hormones (eg parathyroid hormone, calcitonin, insulin, glucocorticoids, 1,25 (OH) ₂ D3) and not a fixed group of growth factors with protein characteristics (osteocalcin, osteonectin, insulin-like) growth factors), see S. Wallach, LV Avioli, JH Carstens Jr., Factors in Bone Formation, Calcified Tissue International 45: 4-6 (1989). The effect of the concentration of hydrogen ions (pH) on metabolic processes in bone regeneration has not been adequately investigated.

Dietz in DE-A-42 40 713 describes the use of a mixture of calcium hydroxide and hoof oil for regenerating collagen after bone injuries. However, this calcium hydroxide and hoof oil formulation suffers from the fact that its stability is very limited as a result of saponification. This may impair the effect of the mixture.

SUMMARY OF THE INVENTION

The invention was therefore based on the intention to provide an improved composition with long-term stability for specific external influencing of bone regeneration processes by stimulating or initiating collagen regeneration.

Surprisingly, it has now been found that by using a composition composed of calcium hydroxide, a dihydric or polyhydric alcohol and a fixed oil of vegetable or animal origin and suitable pharmaceutically acceptable excipients, it is possible to significantly improve the stability of the composition and thus using this composition in bone damage or injury improved collagen regeneration in vivo.

Thus, the present invention relates to a composition comprising calcium hydroxide, a dihydric or polyhydric alcohol and a fixed oil of vegetable or animal origin and suitable pharmaceutically acceptable excipients.

The present invention further relates to a process for the preparation of such a composition by mixing calcium hydroxide, a dihydric or polyhydric alcohol and suitable pharmaceutically acceptable excipients into a fixed oil of vegetable or animal origin.

The present invention further relates to the use of such a composition for the recovery of collagen.

Further, the present invention relates to the use of such a composition for the manufacture of a medicament for inducing collagen regeneration in vivo.

Barium sulfate containing a mixture of calcium hydroxide and hoof oil was used in dentistry as a paste for filling the dental roots (DE-C 29 32 738). Mixtures of carboxylate cement, calcium hydroxide and hoof oil have similarly been used in dentistry as temporary fixation agents for makeshift crowns (DE-C 34 13 864). The role of calcium hydroxide in the previous case is to turn the acidic environment in the dental root canal to alkaline, which causes the elimination of inflammation and the gradual formation of a hard tissue barrier. In the latter case, the prophylactic effect of calcium hydroxide on pulp inflammation is utilized. In both cases, the nail oil serves as a lubricant to ensure that the dental canals are initially simply and completely filled with the active ingredient, calcium hydroxide (and contrast agent, barium sulphate) and, secondly, the curing of temporary fixation agents for temporary dental crowns is slowed by calcium hydroxide. it is also able to penetrate the fine channels of dentine to the pulp and exert its effect there. Neither of the two references gives the slightest indication that the composition of the invention is capable of inducing extensive collagen regeneration as a prerequisite for bone regeneration.

The term preparation, which is novel according to the invention and is used in the following, refers to a pharmaceutical preparation (sometimes also referred to as a mixture) which contains at least one of the aforementioned ingredients. It is particularly suitable for administration to humans or animals for research into collagen regeneration as a prerequisite for bone regeneration.

The ingredients of the composition of the invention are described in more detail below;

Non-solidifying oils of vegetable origin which may be used may contain one or more of the following vegetable oils:

soya, sunflower, rape seed, cotton seed, linseed, castor, palm, palm kernel, coconut and olive oils.

23032004, 04:02:44 Non - solidifying vegetable oils which may be used are preferably non - solidifying oils with high heating stability such as soybean, sunflower and olive oil, and in particular olive oil.

Non-solidifying animal oils which may be used may contain one or more of the following animal oils:

fish oils, animal leg oils and tallow oil.

The animal oils that can be used are preferably animal leg oils, especially hoof oil.

Dibasic or polyhydric alcohols which may be used may be dibasic alcohols such as ethylene glycol, propylene glycol, butylene glycol, pentylene glycol, hexylene glycol, and polyethylene glycols such as diethylene glycol, triethylene glycol, polypropylene alcohol alcohols such as tripropylene glycol, such as dipropylene glycol. arabitol, a5 donitol, xylitol, hexahalic alcohols such as sorbitol, mannitol, dulcitol, or higher polyhydric alcohols.

Preference is given to the use of dibasic and tribasic alcohols such as ethylene glycol, propylene glycol, butylene glycol, pentylene glycol, hexylene glycol and polyethylene glycols such as diethylene glycol, triethylene glycol, polypropylene glycols such as dipropylene glycol, more preferably polyhydric alcohols, tri-ethylene glycol.

Without wishing to be bound by theory, it is believed that dihydric or polyhydric alcohol prevents saponification of vegetable or animal non-solidifying oil. This makes it possible to maintain the mixture for a long time in a kneading or creamy consistency, so that collagen biosynthesis can increase and improve.

The creamy kneadable composition is prepared according to the invention from individual ingredients. The calcium hydroxide is added to the composition in amounts of 1 90% by weight, more preferably 10-70 wt. %, and most preferably 20 60 wt. % based on the total weight of the preparation.

The non-solidifying oil of vegetable or animal origin is added to the composition to produce a creamy, kneadable consistency, preferably in amounts of 9-90 wt. %, preferably 10-60 wt. %, and most preferably 20-40 wt. % based on the total weight of the preparation.

The dihydric or polyhydric alcohol is added to the composition to produce a creamy, kneadable consistency, typically in amounts of 1-40 wt. %, preferably 10-40 wt. %, and most preferably 20 30 wt. % based on the total weight of the preparation.

A preferred embodiment of the present invention relates to the aforementioned composition which additionally contains MgO. For this purpose, MgO may be added in amounts of 1-90 wt. %, preferably 10-70 wt. %, and most preferably

- 60 wt. % based on the total weight of the preparation. It is currently believed that MgO is best added in smaller amounts of 10-20 wt. %.

MgO appears in the bone material as an antacid to counteract the acidic environment in the bone.

The ratio of calcium hydroxide to vegetable or animal non-solidifying oil in the composition according to the invention may be 5/1 to 1/5, preferably 5/1 or 1/1. However, due to the specific circumstances of the trauma of the injury, a deviation from the preferred mixing ratio may be necessary.

If the composition according to the invention is to have a particularly soft and fine consistency, it is also possible to mix petroleum petrolatum therein. Usually it is possible to add white petrolatum in amounts of 1-60 wt. %, preferably in amounts of 10 - 60 wt. %, and most preferably 20-40 wt. % based on the total weight of the preparation.

Although it is not usually necessary to monitor the therapeutic or regenerative processes of the bone radiologically, this may nevertheless be indicated in certain cases. In this case, it is also possible to incorporate barium sulfate as a contrast agent for X-rays into the composition of the invention. However, since the recovery of collagen due to barium sulfate is slightly less good, barium sulfate is added to the composition of the invention, if necessary, in an amount sufficient only to make the composition just radiographically visible.

Depending on its consistency, the application of the composition according to the invention to or in the case of bone damage can be carried out using syringes, spatula or brushes.

There are a variety of uses of the composition of the invention in general surgery, oral surgery, orthopedics, implantology, traumatology and the like, as the composition of the invention can be applied to or damaged bone tissue such as fracture surfaces, drilling, cavity and the like and at a particular application site. immediately induces collagen regeneration

As it is well known that metal fixation agents are used in some relevant medical disciplines, it is advisable in this case to fill in the drilled holes ready to introduce the fixation

Ί of the compositions according to the invention prior to the insertion of the fixation means and only afterwards of the fixation means. In this way, it is possible to counteract the initial osteolysis necessary in such procedures and thus to accelerate the deposition or adaptation of the fixative in or to the surrounding bone tissue and the fixation of the fixative itself in the bone tissue.

In addition, the excess mixture does not interfere in this case, since after the fixation means has been introduced into the bores filled with the mixture, it is expelled or diffuses into spongiosis.

It should be understood that the composition of the invention and its ingredients must be both packaged and applied under sterile conditions.

In a very surprising manner, it has also been shown that the composition according to the invention, even without the support of antibiotics and / or corticoids, acts against the inflammatory reaction caused by bone injury and rapidly causes it to decline. Its simple composition and significant efficacy for collagen regeneration in vivo together with simultaneous inhibition of inflammation make the composition of the invention a composition that will be necessary in the future for bone traumatology.

The following examples are intended to describe the invention in detail.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

First, it is intended to elucidate the interaction between the composition of the invention and the tissue. Thus, data on the distribution of the composition of the invention in bone tissue is a prerequisite to be able to propose a theory about the possible mechanism of action of a drug. Tissues on tissue cultures are therefore more sensitive than those on cell cultures, because only in tissue culture can interactions between cells be studied.

1. Material and methods

1.1. Tissue material

Human osteotomy-derived bone was obtained from hospitals.

Embryonic bone tissue was obtained from chicken embryos (Gallus domesticus) 10-17 days old.

1.2. Tissue culture

The tissue was transferred to the transport medium immediately after removal. Bone fragments of about 2 mm ³ were prepared under sterile conditions and used directly in experiments after weight determination.

Early modified Eagle's minimum essential medium (MEM) with 20 mM Hepes buffer was used for tissue cultures.

Prior to the start of the experiment, 4% fetal calf serum and 1% antibiotic solution (penicillin / streptomycin / amphotericin B) were added to the medium and additionally 1 mM beta-aminopropionitrile, 2 mM sodium ascorbate and 2-10 µg isotopes ( ¹⁴ C- proline). Cultivation was performed in 25 ml Erlenmeyer flasks at 37 ° C in the lowest frequency shaking water bath.

1.3 Determination of respiratory activity

Respiratory activity is a sensitive indicator of tissue metabolic activity. Even minor changes in tissue physiological conditions are reflected in a measurable change in respiratory activity.

A Clark sensor (platinum / silver electrodes in saturated potassium chloride solution) was used to determine respiratory activity. When applying a voltage of 0.8 V to the electrode, the oxygen reduction current is proportional to the partial pressure of oxygen in the measured solution (culture medium). Supply of oxygen-saturated medium where the partial pressure of oxygen drops below a certain value and analytical data is checked by computer.

Bone tissue typically has a respiratory activity of 2-3 µΐ O ₂ x min ^-1 x g ^-1 . Thus, the respiratory activity is in the region of the order of magnitude of the respiratory activity of the remaining muscle tissue. A typical respiratory activity of bone tissue is shown in FIG. 1. The sawtooth of the respiratory curve shown in FIG. 1 results from the fact that when the oxygen partial pressure in the measured solution falls below a certain value, fresh oxygen-saturated medium is supplied.

Fig. 2 shows the average oxygen consumption values from three measurements. Oxygen consumption by embryonic bone tissue (Gallus domesticus) was determined in tissue culture using a Clark scanner. Oxygen consumption is between 3 and 5 μΙ O ₂ x min ¹ x g ^-1 .

Respiratory activity decreases by about 50% over a period that is completely normal for tissue cultures.

1.4. Enzyme tests

The enzyme considered to be closely associated with mineralization of bone tissue is alkaline phosphatase. This enzyme has been characterized some time ago, but discussions are still ongoing on the function of this enzyme in mineralization. Since there is a close association between osteoblast activity and alkaline phosphatase activity, alkaline phosphatase can be viewed as an indicator of osteoblast activity. Elevated levels of alkaline phosphatase activity in blood serum have been observed during skeletal growth in childhood, during bone regeneration and in disorders of bone metabolism.

The alkaline phosphatase activity was determined in the crude extract. For this purpose, 500 mg of tissue was mixed with 1 ml of digestion buffer and cut with a knife. 500 mg of crushing balls were then added and crushed for 20 minutes. After centrifugation, the crude extract was used for measurement.

Alkaline phosphatase was detected by the conversion of p-nitrophenyl phosphate to nitrophenol and phosphate. The nitrophenol that results from hydrolysis is yellow and can therefore be detected by a photometer at 410 nm.

Fig. 3 shows pH dependence on alkaline phosphatase activity. Alkaline phosphatase activity from bone was determined by conversion of p-nitrophenyl phosphate. The maximum activity is at pH 10.5. At physiological pH 7, alkaline phosphatase has only 1% of maximal activity.

1.5 Determination of pH

A composition according to the invention comprising calcium hydroxide, glycerol and hoof oil in a weight ratio of 30 wt. %, 30 wt. % and 40 wt. % based on the total weight of the formulation, or the aqueous calcium hydroxide suspension was overlaid with 30 ml of imidazole / HCl buffer (1 mM, pH 7), after which the pH of the solution was continuously monitored using a pH electrode.

These measurements showed that the mixture of calcium hydroxide and glycerol in hoof oil had substantially different properties from the aqueous calcium hydroxide suspension. Fig. 4 shows that the aqueous calcium hydroxide slurry causes an immediate pH jump to pH 12 and that with the glycerol / calcium hydroxide / hoof oil composition of the invention there is a slow rise above pH 10.

1.6. Determination of collagen

As already mentioned, the major part of the organic substance in the bone consists of collagen, the tissue connecting protein. Bone growth and the regenerative process are associated with the synthesis of new collagen. The synthesis is followed by further intracellular and extracellular processes of collagen. The rate of collagen synthesis in tissue culture can be accurately quantified using a radioactive collagen precursor ( ¹⁴ C-proline). This allows qualitative and quantitative recording of the effect of drugs on collagen synthesis.

The total collagen content is determined by the so-called hydroxyproline test. Collagen is mainly the amino acid hydroxyproline and the hydroxyproline content of other proteins may be neglected. After release of amino acids from the proteins by hydrolysis (16 h at 116 ° C, 22% HCl) and chemical modification (oxidation of 4-hydroxyproline to pyrrole), the total hydroxyproline content of the assay mixture is quantified by specific color reaction with pdimethylaminobenzaldehyde.

1.7. Determination of collagen synthesis rate

Collagen in tissue and medium is obtained by Proff modification (1991) of the Miller and Rhodes method [see EJ Miller and RK Rhodes Methods Enzymol. 82: 33 (1982)]. Through several precipitation steps followed by centrifugation, collagen was quantified by SDS gel electrophoresis and subsequent scintillation measurement (determination of specific radioactivity). To calculate the rate of new synthesis, the collagen content, determined by incorporation of ¹⁴ C-proline, is related to the total collagen content determined by the hydroxyproline assay.

By quantifying collagen biosynthesis, it is possible to investigate the effect of calcium hydroxide products on bone formation.

As stated in 1.7, collagen biosynthesis is quantified by the radiation labeling method of collagen. One of the constituents of collagen fibers is the amino acid proline. A precisely defined amount of labeled ¹⁴ C-proline is added to the culture medium. This proline is compared to newly synthesized proteins during incubation of tissue inkor12. After the collagen has been separated from the other proteins, it is possible to give an accurate account of the degree of synthesis of the new collagen after determining the specific radioactivity.

Collagen is isolated by several precipitation steps followed by centrifugation and SDS gel electrophoresis. In specific precipitation of collagen, the collagen fibers are separated from the other proteins by adjusting the addition of sodium chloride to a suitable salt concentration at which the non-collagenous proteins remain predominantly in solution, but the collagen is excreted as a precipitate. The next centrifugation sedimentes the collagen. In SDS gel electrophoresis, proteins are separated in size-dependent fashion. Proteins migrate in an electric field in a matrix of highly crosslinked polymer (acrylamide). Small proteins migrate rapidly through this matrix because it provides less resistance to small molecules, while large proteins migrate more slowly because their mobility is greatly impeded by the matrix. After staining, the proteins in this gel are visible as so-called bands. In this way, using internal size standards, it is possible to identify proteins based on their size.

For further analysis, for example measurement of radioactivity, it is possible to make proteins available by cutting the bands of interest from the gel.

Collagen extraction:

Tissue culture (see 1.2) was stopped by addition of 3% acetic acid. The collagen that dissolved was precipitated with 2 M sodium chloride at 4 ° C overnight and then recovered by centrifugation (1 h, 24,000 x g, 4 ° C). The sediment was taken up in 10 ml of 3% acetic acid. The newly synthesized collagen present in the tissue blocks was included in the analysis after mechanical disintegration of the tissue blocks. Tissue residues were recovered by centrifugation (1 h, 45,000 x g, 4 ° C). After solubilization, the sediment was fractionated by gel electrophoresis. To control fractionation, the proteins in the gel were stained.

After running, the gel was cut into 5 mm wide bands perpendicular to the direction of migration and the gel fragments were transferred to scintillation vials and counted on a scintillation counter.

Fig. 5 shows a comparison between vital tissue and heat denatured tissue.

This comparison is shown in FIG. 5 based on the distribution of radioactivity in the gel. Collagen, as a relatively large protein, is located in an area 2 cm from the start.

A specific radioactive band can be assigned to a collagen band that is detectable by Coomassie staining.

Fig. 5, tagging 1, femur head, spongiosis, male, 45 years;

difference between synthesis yield between vital and heat denatured tissue (CPM; counts per minute). This shows the distribution of radioactivity in the gel. The collagen band is located about 2 cm from the start. The collagen band is found only in vital tissue, indicating that detectable radioactivity in the gel corresponds to the synthesis of new collagen during incubation.

Vital tissue shows a detectable yield of collagen synthesis, while dead tissue no longer exhibits any metabolic activity. This shows that the detected radioactive collagen is indeed attributable to the synthesis of new collagen in tissue culture and not to the non-specific binding of radioactive proline to bone tissue proteins. Comparative amounts of tissue material (about 100 mg) were used in all experiments.

Other radiation labeled bands of smaller size that possibly are degradation products of collagen can be detected. In particular, degradation of collagen in tissue culture is observed in labeling experiments with an incubation period of more than 4 days. Radiation-labeled proline is also present in the gel to a small extent, which could not be completely removed by precipitation.

The tolerance of bone tissue to weakly alkaline pH is apparent from the parallel experiment shown in FIG. 5. In a parallel experiment, the physiological Hepes buffer in the culture medium (pH 7.4) was changed to a bicarbonate buffer (pH 8.0). As a result, alkalization of the culture medium to pH 8.0 resulted in an immeasurable difference in collagen synthesis versus pH 7.4. Above pH 8.5, any increase in collagen regeneration related to spontaneous collagen regeneration can no longer be observed.

Figures 6 to 9 show the amount of newly synthesized collagen in test batches with various mixtures of the invention with control batches without this mixture or in the absence of a dihydric or polyhydric alcohol. The difference in the amount of newly synthesized collagen in the control batch and in the test batch is expressed as a percentage. 0% means that there is no increase in the amount of newly synthesized collagen compared to control; 100% means that collagen regeneration increased by the composition of the invention twice compared to control.

FIG. 6 shows that in four experiments the collagen synthesis increased between 100% and 120% under the influence of glycerol / calcium hydroxide / hoof oil (composition: 30 wt% Ca (OH) ₂ , 30 wt% glycerol) , 40 wt% hoof oil). This increase is significant because the variation of the experiments in the yield of collagen synthesis ranged from 10% to 20%, while the increases observed under the influence of glycerol / calcium hydroxide / hoof oil are between 100% and 120%. In addition, obviously, there is also an increase in the yield of collagen synthesis compared to the glycerol-free mixture.

FIG. 7 shows that in four experiments the collagen synthesis increased between 100% and 120% under the influence of propylene glycol / calcium hydroxide / hoof oil (composition: 30 wt% Ca (OH) ₂ , 30 wt% propylene glycol) , 40 wt% hoof oil). This increase is significant because the variations of the experiments in the collagen synthesis yield range from 10% to 20%, while the increases observed under the influence of the propylene glycol / calcium hydroxide / hoof oil mixture are between 100% and 120%. In addition, e15 apparently also increases in the yield of collagen synthesis as compared to the propylene glycol-free mixture.

FIG. 8 shows that in four experiments the collagen synthesis increased between 100% and 120% under the influence of glycerol / calcium hydroxide / olive oil (composition; 30 wt% Ca (OH) ₂ , 30 wt% glycerol) , 40 wt.% Olive oil). This increase is significant because the variation of the experiments in the yield of collagen synthesis ranged from 10% to 20%, while the increases observed under the influence of glycerol / calcium hydroxide / olive oil are between 100% and 120%. In addition, obviously, there is also an increase in the yield of collagen synthesis compared to the glycerol-free mixture.

FIG. Figure 9 shows that in four experiments, collagen synthesis increased between 100% and 140% under the influence of glycerol / calcium hydroxide / magnesium oxide / hoof oil (composition; 20 wt.% Ca (OH) 2, 20 wt. % glycerol, 20 wt% magnesium oxide, 40 wt% hoof oil). This increase is significant since the variations of the experiments in the yield of collagen synthesis range from 10% to 20%, while the increases observed under the influence of glycerol / calcium hydroxide / magnesium oxide / hoof oil are between 100% and 140%. In addition, evidently, there is also an increase in the yield of collagen synthesis as compared to a mixture not containing glycerol and MgO.

Example 2

In a stability test in which the following compositions were stored at room temperature and ambient humidity after the kneadable formulation or cream mass were formulated, it was found that maintaining the desired consistency in the presence of a dihydric or polyhydric alcohol could be increased by at least 50% stability compared to with alcohol-free samples.

Mixtures used:

1) Glycerol / calcium hydroxide / hoof oil mixture (30 wt.% / 30 wt.% / 40 wt.

%) and

2) A propylene glycol / calcium hydroxide / hoof oil mixture (30 wt% / 30 wt% / 40 wt%), and

3) Glycerol / calcium hydroxide / olive oil mixture (30 wt% / 30 wt% / 40 wt%)

mixture 1) without glycerol with glycerol kneadable consistency 6 months 12 months creamy consistency 12 months 18 months

mixture 2) without propylene glycol with propylene glycol kneadable consistency creamy consistency 6 months 12 months 12 months 18 months

mixture 3) without olive oil- with olive oil kneadable consistency 6 months 12 months creamy consistency 12 months 18 months

Claims (13)

PATENT CLAIMS WFAN-zooo A pharmaceutical composition comprising calcium hydroxide, a fixed oil of vegetable or animal origin, and optionally pharmaceutically acceptable excipients, and additionally containing a dihydric or polyhydric alcohol. Pharmaceutical preparation according to claim 1, characterized in that the ratio of the volume of calcium hydroxide to the volume of vegetable or animal non-solidifying oil is 5/1 to 1/5. Pharmaceutical preparation according to claim 1, characterized in that the ratio of the volume of calcium hydroxide to the volume of vegetable or animal non-solidifying oil is 1/1. Pharmaceutical preparation according to claim 1, characterized in that the ratio of the volume of calcium hydroxide to the volume of vegetable or animal non-solidifying oil is 5/1. Pharmaceutical preparation according to any one of claims 1 to 4, characterized in that it also contains barium sulfate. Pharmaceutical preparation according to any one of claims 1 to 5, characterized in that it also contains white petrolatum. Pharmaceutical preparation according to any one of the preceding claims, characterized in that it also contains MgO. Pharmaceutical preparation according to any one of claims 1 to 7, characterized in that the dihydric or polyhydric alcohol is glycerol. Pharmaceutical preparation according to any one of claims 1 to 8, characterized in that the vegetable or animal non-solidifying oil is a hoof oil. Pharmaceutical preparation according to any one of claims 1 to 8, characterized in that the vegetable or animal non-solidifying oil is olive oil. A process for the manufacture of a pharmaceutical composition according to any one of the preceding claims, characterized in that calcium hydroxide and a dihydric or polyhydric alcohol are mixed into a fixed oil of vegetable or animal origin. Use of a pharmaceutical composition comprising calcium hydroxide, a dihydric or polyhydric alcohol and a fixed oil of vegetable or animal origin, and optionally pharmaceutically acceptable excipients, for the recovery of collagen. Use of a pharmaceutical composition comprising calcium hydroxide of a dihydric or polyhydric alcohol and a fixed oil of vegetable or animal origin, and optionally pharmaceutically acceptable excipients, for the manufacture of a medicament promoting collagen regeneration in vivo.