DE3427618C2 - - Google Patents

Description

The invention relates to the use of homocarnosine or its pharmacologically acceptable salts to promote healing wounds, ulcerations and inflammatory Processes.

The age of drugs with anti-inflammatory, anti-edematous, antigranular and analgesic effects began with the discovery of the clinical effects of cortisone 1949. Today, these drugs are used only to a limited extent Scope applied.

There are numerous inflammatory processes with the usual ones Antiphlogistica cannot be treated, for example Decubitus, corneal ulcer, cervical erosion and ulcer cruris on the lower legs. The natural or would be ideal spontaneous healing of inflammatory diseases. That stands however experience that spontaneous healings very much are rare.

From the British patent GB-PS 12 52 495 is the use of homocarnosine and its pharmacologically acceptable Salts known to treat bacterial infections.

The invention relates to the use of homocarnosine or its pharmacologically acceptable salts for the treatment of various Types of inflammatory processes. It is based on the surprising finding that homocarnosine or its pharmacologically compatible salts to promote healing of wounds, Ulcerations and inflammatory processes can be used.

Experimental testing for anti-inflammatory activity  with homocarnosine there was an increase in the formation of granulation tissue in the granuloma test.

Homocarnosine is a dipeptide, namely L-histidinyl- γ- amino butyric acid. The connection was established in 1961 by Pisano and Mitarb. isolated from cows' brain extract. The compound is present in the brain in an amount of about 0.007%. Since its discovery, the physiological and pharmacological importance of the compound has remained unknown.

Homocarnosine has the following formula:

It is a white crystalline powder with a melting point of 242-243 ° C and a rotation value = + 23.2 °. Homocarnosine is cleaved in vivo to L-histidine and γ- aminobutyric acid. γ- Amino butyric acid is a commercial product.

Various processes for producing homocarnosine are known. According to the process described in The Journal of Biological Chemistry, Vol. 236, No. 2 (1961), pp. 499-502, a suspension of carbobenzyloxy- γ- aminobutyric acid in methylene chloride is mixed with triethylamine. After the solution obtained has been cooled to -5 ° C., ethyl chloroformate is added and the mixture is kept at this temperature for 10 minutes. Then the solution is quickly mixed with a solution of L-histidine methyl ester. The ester was prepared by adding triethylamine to a suspension of L-histidine methyl ester dihydrochloride in methylene chloride at 0 ° C. The reaction mixture obtained is left to stand at 25 ° C. overnight. Then it is washed with water and 1N sodium bicarbonate solution, dried over sodium sulfate and evaporated to a syrup. The residue is dissolved in methanol and 1N sodium hydroxide solution is added. After standing at 25 ° C. for 3 hours, the solution is adjusted to a pH of 5 with dilute sulfuric acid and evaporated to dryness under reduced pressure. The residue is extracted twice with hot ethanol. Water is added to the ethanol extract. After adding 10% palladium-on-carbon, the mixture is hydrogenated. After the hydrogenation, the catalyst is filtered off and the filtrate is evaporated under reduced pressure. The syrup obtained is dissolved in water and adjusted to a pH of 3 with dilute sulfuric acid. When ethanol is slowly added, the dipeptide sulfate separates out in granular crystals. The product is filtered off and recrystallized from aqueous ethanol. The homocarnosine sulfate melts at 240 ° C with decomposition.

A column filled with the Dowex 50 exchange resin is also used 1N hydrochloric acid and then washed with water until the eluate reacted neutral against thymol blue. Then a 10 percent Solution of homocarnosine sulfate placed on the column and the column was washed with water to separate the sulfate ions, until the eluate reacts neutrally to thymol blue. After that the homocarnosine is eluted with 1N ammonia solution. The eluate is evaporated and the residue is mixed with ethanol and left in the cold. This divides the homocarnosine crystalline.

Examples of salts of homocarnosine are salts of the carboxyl group and salts of the amino group. There are also salts both the carboxyl group as well as the amino group. Examples for salts of the carboxyl group, metal salts such as the sodium, Potassium, calcium, magnesium, zinc and aluminum salt, the ammonium salt and substituted ammonium salts, such as Trialkylamine salts, e.g. B. the triethylamine salt. The salts on  the amino group are derived from inorganic or organic Acids such as hydrochloric acid, sulfuric acid, phosphoric acid, Acetic acid, propionic acid, lactic acid, tartaric acid, citric acid, Succinic acid, maleic acid, benzenesulfonic acid and Toluenesulfonic acid. The salts can be known per se Way by reacting homocarnosine with the corresponding Produce acid or base in stoichiometric amounts len.

The examples illustrate the effectiveness of homocarnosine in the treatment of inflammatory diseases.

Example 1

The increased formation of granulation tissue by homocarnosine was using the usual formalin filter paper method (FFP method) and the quantitative determination of Hydroxyproline examined. Groups were used for the experiments 5 or 6 week old male Wistar rats with a Body weight of about 170 g each used. The experimental animals were shaved on the back and under ether anesthesia a cut of about 1 cm along the dorsal midline guided. Filter paper was in circles with a diameter cut by 6.7 mm. The filter paper was in 7% aqueous Formalin solution dipped. There were four in each rat Filter paper circles under the skin of the right and left scapular areas and under the skin about 4 cm below it Areas inserted, two filter paper discs each on the right and two on the left. The Trial animals were treated with penicillin to give the possibility suppress subcutaneous bacterial infection. The cut was closed with clips. On the second Day after insertion of the soaked in formalin solution The clips were removed from filter paper discs. On the seventh The animals were subjected to ether anesthesia for days Opening the carotid killed. The granulation tissue containing Filter paper disks were removed and wet  weighed. The filter paper discs from the left shoulder blade were used for the quantitative determination of hydroxyproline turns.

An isotonic solution of homocarnosine was made and the animals either subcutaneously or peritoneally once a day injected during 7 days after wound closure. For oral administration, a 15% solution was prepared using was given to a feeding tube. The granulation tissue was separated from the filter paper with a mixture degreased from ethanol and ether and then by autoclaving transferred into a gelatinous preparation. Then that was Preparation mixed with concentrated hydrochloric acid and over Saponified at 105 to 110 ° C overnight. After evaporation was done the residue dried at 105 to 110 ° C. The product was mixed with 0.02N hydrochloric acid and the hydroxyproline content with an automatic amino acid analyzer (Hitachi-835) quantified to the amount of collagen produced to estimate.

Table I

Effect of homocarnosine on the formation of granulation tissue (FFP method)

Example 2

Surgical wound healing by homocarnosine was as follows examined. 6 week old male Wistar rats with a body weight of about 170 g each were considered Laboratory animals used. The rats were over at the back shaved a larger area. Then was under ether anesthesia a cut of approximately along the dorsal median line 4 cm led. Penicillin was given to the possibility suppress a bacterial infection. The cut was made with surgical sutures evenly in three places sewn.

An isotonic solution of homocarnosine was made the daily at 8 locations on both sides of the cut evenly injected subcutaneously for 7 days. As a control served physiological saline. The threads were after 4 days away, and the experimental animals were on the seventh day killed under ether anesthesia. Three pieces of the back skin of 3 cm in length and 1 cm in width were prepared in such a way that the incision line was in the middle of the skin flap. A One side of each skin flap was fixed and the other pulled. The one required to separate the skin at the seam Force (tensile strength) was determined.  

Table II

Effect of homocarnosine on surgical wound healing

Discussion of the test results

Table I shows the results of the granuloma test after FFP method summarized. Homocarnosine in an amount of 10, 30, 50, 70 and 90 mg / kg (i.p.) is effective in education of granulation tissue. The quantitative test for hydroxyproline also shows that its amount increases with increasing Weight amount of granulation tissue increases. In Table II are the effects of homocarnosine on surgery Cutting injuries summarized. The effectiveness is shown by the increased tensile strength of the formed Scar tissue on the seventh day after subcutaneous administration of 0.6 or 6.0 mg homocarnosine / rat.

Inflammation of internal organs is called after the affected one Organ and its pathological condition. Essentially the disease is due to inflammation of the connective tissue in these organs. The beneficial effects of homocarnosine is unspecific for a particular organ. If homocarnosine the diseased area in sufficient concentration  inflammation begins to heal. That's why can homocarnosine for example in the following diseases be effective: corneal injuries, conjunctivitis, wounds various types (surgery), wounds after tooth extraction, Aphthae, angulug infectiosus, allergic rhinitis, Tonsillitis, inflammation of the respiratory tract, burns, Herpes zoster, decubital ulcers, eczema, allergic Dermatitis, ulcers of the stomach, duodenum, Small intestine, large intestine and rectum, archosyrinx, hemorrhoids and ulcer ani, cervical erosions, ulcerations of the Vagina and external genitalia as well as herpes and ulcerative Atheromas.

Homocarnosine works equally well with both oral and parenteral administration. Therefore, for example, it can be used as an injection, Powder, granules, tablet, capsule or tablets or capsules with gastric juice-resistant coating, ointment, Paste, suppository, klysma, inhalation preparation or lozenge are given. Homocarnosine can either be used alone or given together with other drugs. The Dose depends on the type of administration, the preparation and the type and severity of the disease.

Typical recipes for homocarnosine are shown below Given preparations.

Preparation dosage and administration 0.5% homocarnosine injection Solution comes in a dose of 0.1 to 1.0 ml each in the given sick areas. Suppositories contain 20 mg Homocarnosine. The suppositories are in the diseased area introduced. An ointment with a homocarnosine Content of 1% is on the sick Area applied.  

Homocarnosine is practically non-toxic. It is easily soluble in water. That is why easily aqueous isotonic solutions can be used with a content of 0.3, 0.5 or 1.0% below aseptic Create conditions. The solutions can be found at a protective gas can be filled into ampoules. Freeze-dried Homocarnosine can be used in ampoules under aseptic conditions filled and shortly before the injection to a 0.3, 0.5 or 1.0 percent isotonic solution. Homocarnosine is made either by usual galenical methods in the form of a powder, granulate, tablet or capsule processed for oral administration. Examples of auxiliary substances are Binders, such as syrup, gum arabic, gelatin, sorbitol, Tragacanth or polyvinylpyrrolidone, excipients such as lactose, Corn starch, calcium phosphate, sorbitol or glycine, lubricants, such as magnesium stearate, talc, polyethylene glycol, Hydroxypropylmethylcellulose or silica, disintegrant, such as potato starch, and wetting agents such as sodium lauryl sulfate. Tablets can be coated in the usual way. Ointments are made in the usual way by mixing finely powdered Homocarnosine in the desired concentration made with an ointment base. examples for Ointment bases are bleached beeswax, whale wax, anhydrous lanolin, white petrolatum, higher alcohols, Macrogole or plastibase (a hydrocarbon gel ointment base). Furthermore, hydrophilic ointments can absorb water Ointments or their mixtures are produced. If necessary Medicines can contain an oil such as sesame oil, peanut oil or olive oil, a resinous material, glycerin, propylene glycol, a wetting agent, a germicide, a fungicide or an antioxidant can be added. For the production Homocarnosine ointments make the mixture uniform kneaded. Homocarnosine suppositories are in almost manufactured in the same way as the ointments. For example become suppositories by adding antiseptics and homocarnosine to the melted suppository base, thorough Mix, pour into molds and remove the suppo  Sitoria obtained from the mold after solidification.

Below are several recipes for processing Given homocarnosine.

Preparation 1 (solution for injection)

Under aseptic conditions, homocarnosine is in a 0.3 percent, 0.5 percent and 1.0 percent aqueous dissolved isotonic solution. The solutions are packaged in ampoules filled.

Preparation 2 (granules)

Granules are made from the following components:

Homocarnosine 0.2 g lactose 0.34 g corn starch 0.45 g hydroxypropyl methyl cellulose 0.01 g granules 1.00 g

Preparation 3 (ointment)

A 1 percent ointment is made from the following ingredients produced:

Homocarnosine 1.0 g hydrocarbon gel ointment base 99.0 g ointment 100.0 g

Preparation 4 (suppositories)

Suppositories are made from the following ingredients:

Homocarnosine 0.02 g p-Hydroxybenzoic acid ethyl ester 0.00085 g Hosco S-55 suppository base 1.5 g

Homocarnosine is used to produce suppositories and the ethyl p-hydroxybenzoate finely sieved and in small portions in the melt of the suppository heated to 50 ° C Basis entered. It will be one  Get mixture. The mixture is poured into a mold at 38 ° C. After cooling to room temperature the mold frozen in a refrigerator. Then the suppository removed from the mold and placed in a paraffin paper wraps.

Below are the results of clinical studies with homocarnosine preparations in the treatment of tooth extraction wounds, Stomatitis aphthosa, Angulus infectiosus, Herpes zoster, cervical erosion, decubitus, hemorrhoids, Inflammation of the respiratory tract and various Types of dermatitis summarized.

Clinical case No. 1

A 0.5 percent homocarnosine solution for injection was made in given the tooth extraction wound.  

Table III

Clinical case # 2

Patients with aphthous stomatitis received 0.5 percent homocarnosine Solution injected.  

Table IV

Clinical case No. 3

A 0.5 percent homocarnosine paste was in a total dose of 0.8 g three times a day on Angulus infectiosus (Angular cheilitis) applied.  

Table V

Clinical case No. 4

5 percent homocarnosine paste was found in herpes zoster carried.

Patient: woman, 63 years old. Area: The herpes zoster extended from the lower part the right breast to the side of the body. Case history: The patient looked more subjective after 3 days Pain advice. To the sick area 1 g of 5 percent homocarnosine paste was applied. The pain disappeared within about an hour Completely. The same amount was taken five times a day (1 g each) applied. The blisters discolored themselves and dried out and after 7 days could the herpes disease is considered cured will

Clinical case No. 5

Suppositories weighing 1.7 g each with 20 mg homocarnosine were given to patients suffering from cervical erosion.

Table VI

Remarks:

1. The cytological findings were made according to the following classi fication:

Class I: absence of atypical or abnormal cells Class II: atypical cytology, but no signs of malignancy Class III: cytology suspicious, but no clear indication of malignancy (Class III is further divided into Class III _a , Class III _b and Class III _c Class III _a is more benign, whereas class III _c suggests a malignancy, while class III _b lies in between.

2. In all of the clinical cases mentioned above showed as a result of the biopsy that no malignancy Template.

3. The following criteria were used to evaluate the impact of Homocarnosine uses:

-no improvement + no discharge and no bleeding in the eroded area the cervix. The whole area has changed again normalized. ++ The macroscopic examination and the histology confirm that the patient is completely healed (Class I). No discharge and no bleeding. Normal Vagina without further complications.

Clinical case No. 6

1% homocarnosine ointment was applied to a decubital ulcer applied.  

Table VII

Clinical case No. 7

Suppositories weighing 1.7 g each containing 10 mg homocarnosine was used in patients with hemorrhoids give.

Table VIII

Clinical case No. 8

0.1% homocarnosine solution in physiological saline was given to patients suffering from inflammation of the Respiratory tract suffered. The solution was in the respiratory system tracts nebulized.

Table IX

Clinical case No. 9

0.5 percent homocarnosine ointment was used in patients given different types of dermatitis.  

Table X

Claims (1)

Use of homocarnosine or its pharmacological compatible salts to promote the healing of Wounds, ulcerations and inflammatory processes.