DE2949217C2 - Medicines with callus formation and wound healing properties - Google Patents

Description

L-isoleucine

L-leucine

L-lysine (calculated as base)

L-methionine

L-phenylalanine

L-threonine

L-tryptophan

L-valine

L-arginine

L-histidine (calculated as base)

Glycine

L-aanine

L-proline

L-aspartic acid

L-asparagine

L-Cyslin

L-glutamic acid

L-ornithine (calculated as base)

L-serine

L-thyrosine

7.65 mg 1335 mg 8.4 mg 5.7 mg 7.65 mg 6.15 mg

2.7 mg 7.20 mg

13.8 mg

7.8 mg 11.85 mg 2235 mg 1335 mg

1.95 mg 435 mg 0.75 mg 6.90 mg 3.75 mg

1 f \ (\ ma

1.95 mg

The importance of ribonucleic acids, referred to below as RNA, in protein biosynthesis is general known. The genetic information is stored in the DNA and transcribed onto the m-RNA transfer. In the translation phase, the information is transmitted with the participation of the ribosomes and through specific interaction of m-RNA and the

t-RNA translated into the corresponding protein structure In the current experiments, heterologous Organ-specific RNA worked with the preparation of Bone Regenerated from VC 5 of the laboratory Prof. Dyckerhoff from Cologne.

Kallikrein is a kinin-releasing protease whose inactive precursor in the plasma due to contact with the External surfaces - also wound surfaces - activated It has already been established that kallikrein dip This stimulates the DNA synthesis of resting cells initiates and intensifies the mitotic activity DNA synthesis and mitosis is said to depend on the increase in cell membrane permeability by bradykinin, of the extracellular calcium concentration and the altitude the intracellular cAMP level. During the investigation of the fracture healing it was found that amino acids, ribonucleic acids and kallikrein individually or in combination are a cause a certain acceleration of the callus formation. However, the exogenous supply of cAMP is from It is of crucial importance, as this significantly accelerates and improves healing could be achieved. About the combination of cAMP with amino acid mixtures, then about the Combination of cAMP with amino acids and RNA up to the application of cAMP, amino acids, RNA and Kallikrein showed a gradual, significant improvement in fracture healing processes under all conditions, the optimal toilet in the latter mixture. t reached.

The agent according to the invention is preferably in liquid form as an injection or infusion. . used because oral administration is not possible / ¹ S starting substances the commercially available ones can be used. The dosage for cAMP is generally 4-6 mg / kg body weight and preferably about 4.2 mg / kg; the amino acid mixtures are used in a dosage of about 100-200 mg / kg body weight and preferably 150 mg / kg body weight. Ribonucleic acids are used in amounts of about 5-10 mg / kg body weight and preferably about 8 mg / kg body weight, the dosage for kallikrein is about 30-80 and preferably 50 KE / kg body weight

The invention is explained in more detail below with reference to the examples:

example 1

In all pharmacological investigations, 4-month-old males were used as test animals New Zealand rabbits with an average weight of 3000 g used. Each test series included 12 animals. 12 other rabbits served as control animals, for a total of 24 rabbits.

Animals were excluded from the tests with infection of the breaking point, with intercurrent illnesses or different fracture types, so that the number of animals in the individual series low t_ / fllWI iibtllbUb. BUI ΤΙΙ, ΐΟΙ *

Test procedure

Under anesthesia, the right tibia was fractured in the middle of the diaphyses over a wedge and the fragments through a Kirschner wire inserted intramedullary from the head of the tibia fixes the fracture shape and position of the Kirschner wires were checked radiographically.

During the 2 weeks of the experiment, the test substances were given to the rabbits in various dosages and application SL. ien injected The amount of Dose was determined either by the optimal sizes given in the literature or when this information was missing, obtained in extensive preliminary tests with series of different concentrations

In previous attempts, the 14th day after the fracture was found to be a good time to examine the Proven healing results. Physiological fracture healing was in the 4-month-old rabbits and in this experimental set-up, like the controls show on 20.-21. Day post fract completed. In order to be able to demonstrate the effects of biochemical agents on fracture healing, the Try the time of the physiological fracture healings shortened by '/ i and the 14th day post fracture was chosen for the examination

The test animals were killed painlessly on this day by an overdose of barbiturate. The TiDia was dissected free and the Kirschner wire An X-ray was taken of each tibia, the osseous structure assessed and the Localization of the fracture point determined

To objectify the results, the bones were bent in a bending machine of the State Factory Testing Office of the Hamburg Engineering School until they were refractured cm diameter with plaster necessary. The Gipsveranke- ^J "rung of the bone ends in the metal sleeves was stable. During the bending test, there was in any case to a solution or deformation of the anchorage. The prepared bone was clamped in the holders of the bending machine. The measured clamping points were each 6 cm from Frakturspait removed Thus the lever arm of the acting bending force was uniformly defined

When testing the Callus stability, the

Load in pond (ρ) and the bend in mm is phonetic

^{4 (>} recorded The flexural rigidity of the fracture points was derived from these values using the following formula

Flexural stiffness = force (ρ) * force arm (cm)
Bend (mm)

The measured quantities force and bending still remaining in the equation, which are recorded for each bone enabled the graphic representation of the

»Breaking process. A characteristic curve was created by plotting the bending value on the abscissa and the values for force on the ordinate. It consisted of a more or less steep ascent phase and a terminal flattening. The ascent phases of the curves differ markedly from one another in the angle of incline or ascent. The terminal flattening indicated the beginning of the callus break and was similar for all curves.
There was a direct proportion between the angle of rise of the curves and the breakthrough of the fractures. When examining non-fractured bones, the ascent angle reached values between 80 ° and 88 °.

While in the physiological fracture healing on the 14th day a mean value of the angle of increase at a degree diameter of 2 mm of about 45 * was found among the the above-mentioned test conditions for treatment

with heterologous bone RNA in a dose of 2.4 mg daily an angle of increase on the average of 56 °. when using amino acid mixtures in the manner described in a dosage of 3 ml per day a mean value of the angle of about 69 ° and when using kallikre; n in a dosage of 3 times a day 501. E. one of about 63 °

After injecting the test animals according to the test conditions described above with 12.5 mg daily cAMP, the mean value of the angle of increase after 14 days was about 71 °.

Example 2

Under the same test conditions, the test animals received 12.5 mg of cAPM, 3 ml of the specified amino acid mixture, 2.4 mg of bone RNA and 150 KE FCallikrein daily. The ^': -lwert the rise angle was to i4 """.ige · wa 79 ° A comparison of the values of the fräkturierten and treated tibias with dene.?" there was no longer any significant difference in the unbroken tibiae of the other extremities of the same animals with this treatment; the broken and treated bones had reached the resilience of the unbroken

The treatment with the agents according to the invention can therefore reduce the fracture healing time by about • Shorten / 3.

Claims (1)

Claim: Medicines to promote callus formation, characterized by a content of cyclic adenosia phosphate, essential amino acids, Bone ribonudic acids and kallikrein. The invention relates to a medicament with callus formation and wound healing promoting effect. The first investigations into the Cailus formation have been carried out since as early as the 17th century Attempts have been made from many sides to include bone bone healing and wound healing affect physical, biological and biochemical agents. The best success in fracture treatment is based on taking physical laws into account. From a biological point of view, autoplastic bone and cancellous bone transplantation gained particular advantages Importance for bridging fractures. However, attempts have long been made to influence the healing of fractures and wounds with drugs For a long time, various calcium salts have preferably been used for this purpose, often with the addition of Ascorbic acid, vitamin A or vitamins of the D series are used. Disadvantages of these known products is that an effect, depending on the general condition and the level of desire of the psiiemen, is often not or only m to a small extent can be determined and that in combinations with vitamins of the D series in the often necessary administration over a longer period of time the known side effects of the D vitamins in Appearance. In addition to these most commonly used substances, numerous others have also been found on theirs Effectiveness in callus formation investigated, for example lead salts as direct calcifying agents or Ribonucleic acids. Even with these substances, the effectiveness is different and there are side effects to be expected. The invention was therefore based on the object of providing new methods for influencing the drug Develop fracture and wound healing. To solve the problem, a drug that promotes callus formation and wound healing is proposed, that by a content of cyclic adenosine monophosphate, essential amino acids. Is labeled bone ribonucleic acid and kallikrein. Surprisingly, it has now been found that the agents according to the invention can achieve a significant proliferative effect and that the fracture healing time can be shortened by about a third. Since the Caüus is stimulated by means of the invention, the organic Auibäu, a proloferativer effect is likely not only in fracture healing, but also in wound healing. Cyclic adenosine monophosphate, hereinafter referred to as cAMP for short, is carried out enzymatically in the body the adenyl cyclase bound to the cell membrane is formed from adenosine triphosphate and a phosphodiesterase degraded to adenosine 5'-monophosphate. Under the action of adenyl cyclase it comes to an increase in the intracellular concentration of cAMP. that as an allosteric activator on enzymes acts, the secondary enzymes through ^ h Inhibit or increase the activity of phosphorylation. The organ specificity of those acting via cAMP 'Hormones come about because only the successor organ has the hormone-specific receptor Because the hormone specificity in this mechanism ends at the cell receptor, and the one triggered by it Secondary reactions for all hormones on the principle "· The cAMP effect expire, the cAMP is also called second messenger means an increase in the cAMP content of the cell increases the DNA synthesis and mitotic activity, and cAMP also favors the Emptying of cellular calcium deposits. The increase in '"' Intracellular calcium concentration causes on the one hand a regulation of calcium-dependent; - reactions of the metabolism, on the other hand also an inhibition of the adenyl cyclase,
From DE-OS 27 32 179 it is already known to use cAMP for the treatment of irritation and inflammation of the skin and mucous membrane, but here cAMP acts as a histamine blocker The indispensability of essential amino acids was shown in animal experiments in the case of a lack of the corresponding Amino acid mostly through observation of a growth inhibition, i.e. a disturbance of the enchondral Ossification noted There is a high percentage of proline and collagen in the connective tissue Hydroxyproline; Hydroxylysine comes almost exclusively in this tissue. These amino acids are a characteristic part of the organic bone matrix. Invention geniuses are therefore amino acid solutions which are high in essential amino acids, as found in numerous Preparations used in human medicine for protein substitution are available. During the examinations the best results were obtained with an amino acid mixture containing the following in 3 ml Quantities contained: