ANTI-INFLAMMATORY, ANTIBACTERIAL, WOUND- UND BURNHEALING ACTION

The invention is referred to the field of biologically active compounds and may be used to produce vulnerary medicinal preparations for treatment of skin damages, wounds, burns and skin diseases, method of its production.

In spite of great variety of proposed methods and preparations the issue of treatment of wounds, burns and other skin damages remains topical. In most cases local medicinal treatment is preferred in wound healing (B.K. Gavrilyuk. Nanocomposite regeneration systems for wound healing/ B. . Gavrilyuk, V.B. Gavrilyuk - (Biophysics of complex systems)// Biophysics. - 201 1. - V. 56, No. 6. - P. 1 138-1 141 ; Lloyd EC, Rodgers BC, Michener M, Williams MS. Outpatient burns: prevention and care. Am Fam Physician. 2012 Vol.85(l), P.25-32.; Profyris C, Tziotzios C, Do Vale I. Cutaneous scarring: Pathophysiology, molecular mechanisms, and scar reduction therapeutics Part I. The molecular basis of scar formation. J Am Acad Dermatol. 2012, Vol.66(1), P.1-12.; Elliot D, Sierakowski A. The surgical management of painful nerves of the upper limb: a unit perspective. J Hand Surg Eur Vol. 201 1 Vol.36(9),P.760-770). However, it should be noted that in this case the efficiency of proposed therapy depends not only on correct choice of medicinal preparation, but also on the form in which it should be applied (Endorf FW, Ahrenholz D. Burn management. Curr Opin Crit Care. 201 1 Vol. 1 (6), P.601-605). Anti-inflammatory medicinal preparations subdivided into steroidal and nonsteroidal ones are widely used. Nonsteroidal anti-inflammatory preparations reduce exudation, suppress tissue proliferation, analogize and reduce temperature. Their disadvantage is high toxicity. Steroidal anti-inflammatory preparations are similar to glucocorticoids. By the efficiency of anti-exudate effect they predominate over nonsteroidal antiinflammatory preparations several times, but they do not have analgetic and antipyretic action, and in case of long-term application they cause disorder of endocrine profile of the organism. Also antimicrobial preparations are applied, but they facilitate allergization of the organism, development of stabile wound microflora and dysbacteriosis. Application of enzyme preparations (proteinase, trypsin, chymotrypsin, amino acids and other) is limited by their efficiency only during the first phase of wound process and necessity to combine them with other vulnerary medicinal preparations. Complex action on wound is provided by medicinal preparations of vegetable origin by virtue of content of the complex of flavonoid compounds and vitamins, but they do not have analgetic action (Mendonca Machado N, Gragnani A, Masako Ferreira L. Burns, metabolism and nutritional requirements. Nutr Hosp. 201 1 Aug;26(4):692-700; Lloyd EC, Rodgers BC, Michener M, Williams MS. Outpatient burns: prevention and care. Am Fam Physician. 2012 Vol.85( 1 ), P.25- 32). Vulnerary and anti-inflammatory preparations in a form of creams and ointments containing active agent, oily base (mixture of lanolin and vaseline) and additives are well-known. However, vaseline-lanolin base in these medicinal preparations prevents normal gas exchange reducing wound healing activity of the preparation, especially in case of smoldering processes (Profyris C, Tziotzios C, Do Vale I. Cutaneous scarring: Pathophysiology, molecular mechanisms, and scar reduction therapeutics Part I. The molecular basis of scar formation. J Am Acad Dermatol. 2012, Vol.66(l ), P. l -12.).

Efficient forms of vulnerary preparations are gels. Polysaccharides, such as galactomannan or glucomannan or alginic acids or alginic acid salts or agar-agar or hyaluronic acid or gelatin or pectin or aerosol or zeolites or collodion or 1,2 propylene glycol, may be used as gelatinization agents (Elliot D, Sierakowski A. The surgical management of painful nerves of the upper limb: a unit perspective. J Hand Surg Eur Vol. 201 1 Vol.36(9),P.760-770).

Vulnerary medicinal preparation "Methyluracil liniment" (RF patent No. 2007997, prior, dated 08/07/92, A 61 K 9/06) containing β -form of methyluracil as an active agent, and castor oil as a base, is well-known. The preparation has increased wound-healing activity due to high bioavailability of the pharmaceutical form. However, the active agent in this medicinal preparation does not have anticontagious and interferon inducing properties, which does not allow healing of infected wounds.

The object of this invention is to create the preparation devoid of abovementioned disadvantages; herewith the technical result achieved during implementation of this object consists in production of efficient anti-inflammatory, vulnerary and analgetic preparation for external use.

The indicated technical result is achieved in the following way: original peptide substance Tyr-Pro- X where X - OH, NH ₂ , OC¾, OC2H5 is used as active agent in the following types of pharmaceutical forms: ointments, gels, liniments, sprays, suppositories, plaster, orally disintegrating tablets and drops.

In particular, application of dipeptide with general formula Tyr-Pro-X where X - OH, NH ₂ , OCH ₃ , OC2H5 as an anti-inflammatory, antibacterial, vulnerary, regenerative, analgetic and burntreating preparation, as well as the pharmaceutical form containing dipeptide with general formula Tyr-Pro- X where X- OH, N¾, OCH3, OC2H5 as active agent, applied as anti-inflammatory, antibacterial, vulnerary, regenerative, analgetic and burntreating product, is claimed;

preferable but unessential embodiments of the pharmaceutical form provide its embodiment in a form of one of the following pharmaceutical forms: ointment, gel, liniment, spray, suppository, plaster, orally disintegrating tablets and drops.

According to the present application, peptide compounds, their properties and method of their production are not described in the literary sources.

Synthesis of compounds. Tyr-Pro-X was synthesized with application of tetrabutylammonium salts (TBA) and pivaloyl chloride (PivCL). Derivatives of amino acids D; L; DL were used for synthesis. Evaporation of solutions was performed in the vacuum evaporator at 40°C. Fusion temperatures determined on the warm stage Boetiys are given without correction. Control over the synthesis and individuality of obtained compounds were checked by means of thin-layer chromatography on plates with silica gel produced by Silufol (Czech and Slovak Federal Republic) sprayed with the ninhydrin solution in solvent systems:

1 - hexane:acetone (3:2);

2 - ethyl acetate: acetone: 50% acetic acid (2: 1 :1);

3 - benzene:ethanol (8:2);

4 - chloroform:methanol:ammonia (7:2.5:0.5);

5 - chloroform:methanol:ammonia (6:4: 1);

6 - chloroform:methanol:acetic acid (42:7: 1);

7 - acetone: benzene: acetic acid (2: 1 : 1);

8 - chloroform: methanol:ammonia (8: 1.75:0.25);

9 - chloroform:methanol (9:1);

10 - ethanol:ammonia (7:3).

11 - isopropanol: formic acid:water (20: 1 :5).

Hereinafter solvent system is indicated in brackets after Rf.

All solvents were dehydrated as appropriate. Fusing points were not corrected. Column chromatography was performed with application of carriers Silica gel L-40/100 m and Silasorb 600 [LC] produced by Chemapol (Czech and Slovak Federal Republic).

Authorized abbreviation: Ac - acetate, Boc - t-butyloxycarbonyl group; OBT - 1-oxybenztriazole;

D F - dymethylformamide; DCC - dicyclohexylcarbodiimide; PivCI - pivaloyl chloride; TBA - tetrabutylammonium; TEA - triethylamine; OSu - N-hydroxysuccinimide ester, DCC - N,N- icyclohexylcarbodiimide; OMe - methyl ester; DiBoc - tert-butyloxycarbonyl, TLC - thin-layer chromatography.

Production of dipeptide Tyr-Pro.

1. 2.83 m (1.326 mol) of 13% TBA were added to Pro 150 mg (1.83 mmol), then it was evaporated once with ethanol, once with isopropanol, once with benzene, and then it was cooled up to 0°C.

2. When cooling 634.5 mg (1.326 mol) of dry DiBoc-Tyr-OSu were added to cooled TBA Pro solution in 10 ml of abs. ethyl acetate, stirred within 1 hour at room temperature on magnetic stirrer; the process of reaction was observed by means of TLC in the system (7).

Upon completion of reaction the solvent was evaporated, water was added, then it was acidified with NaHSC (five-fold excess) up to pH 3, extracted 5 times in the amount of 50 ml with ethyl acetate. Combined ethyl acetate fractions were rinsed with water, 10% solution of KHSO4, water, then they were dried over MgSC>4, evaporated, precipitated from ester with hexane (twice). The precipitation in a form of powder was dried in vacuum.

The output was 1.32 g (2.76 mmol).

Rf = 0.382 (7).

The following examples illustrate the efficiency of application of claimed new peptide compound as an active agent in anti-inflammatory, vulnerary and analgetic preparations for external use, as well as variants for production of such preparations. It is understood that, although preferable embodiments are disclosed in detailed description and specific examples, they are represented only for illustration purposes, as various changes and improvements that do not go beyond the limits of concept and bounds of the invention will be evident for the specialist in this technical sphere from their description and claims.

Example 1. For 100 suppositories: 55.0 g of molten mixture of PEG 1500 and PEG 400 with 5.0 g of emulgent are added into solution of the substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH3, OC2H5 with concentration c=l -50 mg/ml, then it is stirred for 10 minutes. 275 g of previously melted hydrophobic base (witepsol) with 15.0 g of stabilizer (Nipagin) are gradually added to the mixture and stirred for 30 minutes. Obtained mixture is poured into previously cooled suppository forms and placed in the refrigerator at -4°C. The preparation is applied as suppositories. 1 suppository contains 1 -50 mg of the substance with peptide nature Tyr-Pro-X where X - OH, NH ₂ , OCH _3; OC2H5.

Example 2. For 100 suppositories: 275 g of previously melted diphilic base (Suppocire AP) with 15.0 g of the stabilizer (Nipagin) are gradually added to the solution of the substance with peptide structure Tyr-Pro-X where X - OH, N¾, OCH3, OC2H5 with concentration c=l-50 mg/ml and stirred for 30 minutes. Obtained suspension is poured into previously cooled suppository forms and placed in the refrigerator at -4°C. The preparation is used in a form of suppositories. One suppository contains 1-50 mg of the substance of peptide nature Tyr-Pro-X where X - OH, NH2,

Example 3. For 100 g of ointment: solution of the substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH3, OC2H5 with concentration c=l-50 mg/ml is introduced into 85.0 g of the melted mixture of PEG 1500 and PEG 400 with 5.0 g of stabilizer (Nipasol) and stirred for 30 minutes. The preparation is used in a form of ointment containing 1 -50 mg of the substance of peptide nature Tyr-Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ .

Example 4. For 100 g of ointment: solution of the substance with peptide structure Tyr-Pro-X where X - OH, N¾, OCH ₃ , OC2H5 with concentration c=l-50 mg/ml is introduced into 40.0 g of the melted mixture of PEG 1500 and PEG 400 with 5.0 g of emulgent, then stirred for 10 minutes. gradually added to the mixture and stirred for 30 minutes. The preparation is used in a form of ointment containing 1-50 mg of the substance of peptide nature Tyr-Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ .

Example 5. For 100 g of gel: solution of the substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH3, OC2H5 with concentration c=l -50 mg/ml is introduced into 85.0 g of the melted mixture of PVP 12600 and PVP 8000 with 5.0g of stabilizer (Nipasol) and stirred for 30 minutes. The preparation is used in a form of gel containing 1-50 mg of the substance of peptide nature Tyr- Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ .

Example 6. For 100 ml of spray: solution of the substance with peptide structure Tyr-Pro-X where X - OH, NH2, OCH3, OC ₂ ¾ with concentration c=l -50 mg/ml is introduced into the mixture containing 20 mg of propylene glycol, 20 mg of PEG 1000 cetostearyl ester, 5ml of purified water and 50 ml of rectified ethanol, then it is stirred for 10 minutes and placed into polypropylene bottle with dispensing pump and nozzle. The preparation is applied in a form of spray containing 1 -50 mg of the substance of peptide nature Tyr-Pro-X where X - OH, N¾, OCH ₃ , OC2H5.

Example 7. For 100 plasters: 5 g of the substance with peptide structure Tyr-Pro-X where X - OH, NH2, OCH ₃ , OC ₂ H ₅ are introduced into the mixture consisting of 9.72 g of poli(DL-lactide) and/or poly(DL-lactide-co-glycolide) with molal weight 50,000 Da and correlation 50:50 with 1.0 g of stabilizer (sodium benzoate) in 60 ml of chloroform. The mixture is thoroughly stirred and poured in a thin layer, after that it is dried with warm air up to complete elimination of organic solvent. Obtained film is combined with tissue base. The preparation is used as a plaster containing 1 -50 mg of the substance of peptide nature Tyr-Pro-X where X - OH, NH ₂ , OCH3, OC2H5.

Example 8. For 100 orally disintegrating tablets: tablets were produced by means of direct compression of the mixture containing 1 g of the substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH3, OC2H5, and 20 g of combined filler "Ludiflash" on the tableting press.

Example 9. For 100 orally disintegrating tablets: tablets were produced by means of wet granulation of the tableting mixture containing 1 g of the substance with peptide structure Tyr-Pro- X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ and 20 g of combined filler "Ludiflash".

Example 10. For 100 orally disintegrating tablets: tablets were produced by means of dry pelletizing of the tableting mixture containing 1 g of the substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ and 20 g of combined filler "Ludiflash".

Example 12. For 1 ml of solution for drops the solution of the substance with peptide structure Tyr- Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ with concentration c=0.0l -10 mg is introduced into the mixture containing sodium phosphate 0.1-5.0 mg, sodium dihydrogen phosphate 1.0-10.0 mg, sorbitol 1.0-25.0 mg, benzalkonium chloride 0.1 -3.0 mg, lavender oil 1 .0-30.0 mg, purified water up to 1 ,0 ml. The preparation is used in a form of nasal drops containing 0.01 - 10 mg of the substance of peptide nature Tyr-Pro-X where X - OH, NH ₂ , OCH ₃ , OC2H5.

Example 13. Study of the preparation for external use containing original substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH3, OC2H5 in the formalin test in order to identify analgetic and anti-inflammatory activity.

Male outbred rats were placed into individual plastic boxes. Acute inflammatory response (oedema) was caused subplantarly (under plantar aponeurises) by means of introduction of 50 mcl of 5% formalin. Number of pain reactions was registered (tapping on the floor with a paw, gnawing of a paw, etc.) within first 5- 10 minutes - 1 phase (action on primary pain afferents) and from 30 ^th to 50 ^th minute - II phase (pain occurred as a result of inflammatory response) (Bannon et al., 1998). Reliable reduction of the number of pain reactions in the test group in relation to the control group was considered to be the criterion for analgetic effect.

Test specimens (specimen 1 - ointment 5%, specimen 2 - gel 10%) were applied to the paw simultaneously with introduction of formalin in male outbred rats with weight of 200-300 g. Dermatic preparation without active agent was applied to control animals. Obtained results are represented in table 1.

Table 1

- reliability in comparison with the control at PO.05.

From table 1 it appears that the preparation for external use containing original substance with peptide structure Tyr-Pro-X where X - OH, N¾, OCH3, OC2H5 reduces algesthesia during the I phase of formalin pains characterizing the impact on primary afferents and has anti-inflammatory action during the II phase caused by the development of inflammation.

Therefore, the preparation for external use containing original substance with peptide structure Tyr- Pro-X where X - OH, N¾, OCH3, OC2H5 has analgetic and anti-inflammatory action in formalin test. Example 14. Study of the preparation for external use containing original substance with peptide structure Tyr-Pro-X where OH, NH ₂ , OCH ₃ , OC2H5 when molding thermal burn.

In order to mold thermal burn standard method was used (H.I. ochtygov, On thermal burn reproduction methods, L.: Meditsyna, 1964, page 86) on chinchilla rabbits with weight of 2.8-3.2 kg. Burns were made on the hip. Hair was removed from the area of animal hip, and ceramic sagger 1.5 cm in diameter, heated in the muffle furnace up to 600°C, was applied to skin for 5 seconds. Then studies were carried out in three animal groups. In the first group (control) no treatment was performed. In the second group (test) treatment was performed with the preparation for external use in a form of ointment without active agent produced according to the prototype. In the third group (test) treatment was performed with the preparation for external use in a form of ointment with concentration of active agent 5.0 % wt. In the fourth group (test) treatment was performed with the claimed preparation for external use with concentration of active agent in a form of gel 10.0 % wt. Animals were inspected on a daily basis; condition and dimensions of burn wound were determined. On the 7 ^th , 14 ^th and 28 ^th day burn wounds were totally dissected out together with healthy skin area and underwent microscopic examination.

In the control group dry coagulation necrosis area clearly separated from the surrounding undamaged tissue was formed on the place of thermal burn immediately after it was made. After 1 day the demarcation line of damaged epidermis 3-4 mm in dimensions formed around this area. Starting from the 2 ^nd day fibrinous film formed on the surface of burn wound; it became thicker and darker during the first week. During next 7 days perilesional hyperemia was observed, and inflammatory swab was formed. Herewith dimensions of lesion focus reduced considerably. By the 14 ^th day lesion focus reduced due to the ingrowth of epidermis in it from the periphery in a form of tongues. By the 28 ^th day burn wound healed completely.

In the 2 ^nd group of rabbits (usage of the preparation for external use without active agent) healing of burn wound took place in a shorter time than in the 1 ^st group; however, during the entire period of treatment local inflammation was significant. The form of burn wound changed due to uneven ingrowth of epidermis. By the 21 ^st day the wound completely epithelized from its edges, and by the 28 ^th day it was completely healed.

In the 3 ^rd group (external use of the medicinal preparation with the concentration of active agent 5.0 % wt) good results were registered. During first seven days heavy local inflammatory response was observed, but on the 14 ^th day inflammation completely disappeared, and burn wound epithelized due to formation of regenerating epithelium from the edges. Complete union of epithelium took place on the 21 ^st day of treatment, 7 days earlier than in the 2 ^nd group (prototype) and two weeks earlier than in the control group. No epulosis or cicatrization was observed. In the 4 group (external use of the medicinal preparation with the concentration of active agent 10.0 % wt) the best results were observed. Heavy local inflammatory response was observed during first five days only, but on the 10 ^th day the inflammation completely disappeared, and burn wound epithelized. Complete union of epithelium took place on the 18 ^th day of treatment, 10 days earlier than in the 2 ^nd group (prototype) and control groups. No epulosis or cicatrization was observed. Therefore, the preparation for external use containing original substance with peptide structure Tyr- Pro-X where X - N¾, OCH3, OC ₂ H5 shows high vulnerary, anti-inflammatory and anticontagious efficiency and reduces terms for treatment of infected wounds and burns.

Example 15. Studying of the preparation for external use containing original substance with peptide structure Tyr-Pro-X where X - OH, N¾, OCH3, OC2H5 in molding of dermal wound.

Dermal wound was molded on chinchilla rabbits with weight of 2.8-3.2 kg. In order to mold the wound the area of soft tissues on rabbit hips was shaved, and the incision 1 cm in depth and 5 cm in length was made with lancet. After that the study was performed in three animal groups. In the first (control) group the treatment was not performed. In the second (test) group treatment was performed with the preparation for external use in a form of ointment without active agent produced according to the prototype. In the third (test) group treatment was performed with the preparation for external use in a form of ointment with the concentration of active agent 5.0 % wt.

The efficiency of preparations in test and control groups was evaluated according to crust rejection time, wound cleansing of purulonecrotic masses, time of granulation tissue onset in the wound and initiation of acentric epithelialization according to dynamics of inflammatory process regress, as well as according to dynamics of wound bacterization.

In the control group abscess with infiltration into surrounding tissue was formed. On the 6 ^th day crust rejection and acentric granulation tissue occurred, and by the 12 ^th days of the course of treatment first signs of epithelialization occurred. Complete debridement of wound took place by the 21 ^st day.

In the second group of rabbits pyogenic serous drainage in the wound was absent. Primary crust on the wound did not prevent formation of granulation. Crust rejection and first signs of granulation were observed on the 6 ^th day of treatment. Occurrence of acentric wound epithelialization was registered on the 12 ^th day of treatment, and complete debridement of wound took place by the 21 ^st day of treatment.

The best results of treatment were observed in the third group (preparations for external use containing 5% of the substance). Neither purulent discharge nor serous drainage from the wound was observed; inflammation was not observed; wound was "dry", formation and growth of granulation tissue (active at the periphery and insular in the center of the wound) were observed. Priority development of granulation tissue facilitated earlier epithelialization and complete debridement of wound that took place in this group by the 9 ¹ day of treatment; complete healing of the wound took place by the 15 ^th day of the course of treatment. No epulosis or cicatrization was observed.

Therefore, the preparation for external use containing original substance with peptide structure Tyr- Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ has vulnerary effect.

Example 16. Study of the preparation for external application containing original substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH3, OC2H5 in the test of neurogenic pain syndrome caused by sciatic nerve transection as a mold of neuropathic pain in order to identify analgetic and anti-inflammatory activity.

The experiments were carried out on male white non-pedigree rats with weight of 200-250 g. Neurogenic pain syndrome in male outbred rats was created by means of sciatic nerve transection at the level of popliteal space above the place of its trifurcation on n. tibialis, n. perone s and n. suralis and its further ligation in order to prevent regeneration. Development of pain syndrome was determined by occurrence of signs of autotomies on the paw which underwent surgery. Intensity of autotomies was evaluated according to 11 -point scale: 1 point - damage of 1 claw; 2, 3, 4 and 5 points - damage of 2, 3, 4 and 5 claws; 6 points - damage of maniphalanx of one finger; 7, 8, 9, and 10 points - damage of maniphalanxes of 2, 3, 4 and 5 fingers correspondingly; 1 1 points - damage of instep bones. Pain syndrome development dynamics in rats of test and control groups was observed within 25 days (G.N. Kryzhanovsky et al., 1991).

In the day of sciatic nerve transection and during next 10 days after surgery the preparation for external use in a form of ointment without active agent was applied to animals of the control group, and animals of the test group received the preparation for external use in a form of ointment with active agent in a dose of 10 mg kg within the same period.

In the control group of rats development of autotomy after sciatic nerve transection occurred in 50% of animals on the 6 ^th day after surgery. The intensity of pain reactions made 3.2±0.9 points. On the 9 ^th day of introduction pain reaction was observed in 60% of animals, corresponding to the intensity of 4.5±2.4 points. On the 18 ^th day pain reactions developed in 70% and made 6.5±3.1 points.

Application of the preparation for external use without active agent did not prevent development of autotomy after sciatic nerve transection (40% of animals on the 6 ^th day after surgery). The intensity of pain reactions made 2.7±1.9 points. On the 9 ^th day of introduction pain reaction was observed in 50% of animals, corresponding to the intensity of 3.6±2.6 points. On the 18 ^th day pain reactions developed in 60% of animals and made 5.2±2.4 points.

Application of the preparation for external use with active agent prevented development of pain syndrome. On the 6 ^th and 9 ^th day no pain reaction was observed. On the 13 ^th day of the experiment and on the 5 ^th day after cancellation of introduction of the compound pain reaction occurred in 10% of animals with medium intensity of 1.0±1.0 points, and on the 18' day - in 10% of animals, medium intensity of reaction made 2.0±1.0 points.

Therefore, the preparation for external use containing original substance with peptide structure Tyr- Pro-X where X - OH, NH ₂ , OCH3, OC2H5 shows analgetic and anti-inflammatory action during the test of neurogenic pain syndrome caused by sciatic nerve transection.

Example 17. Study of the preparation for external use containing original substance with peptide structure Tyr-Pro-X where X - OH, N¾, OCH3, OC2H5 in the patient E, 68 years old. In the patient pyogenic acne on the face near the nose were diagnosed. The preparation for external use in a form of ointment was applied to all damaged skin areas. On the next day after single application light crust formed; it could be easily eliminated. No traces of wounds remained on the face.

Example 18. Study of the preparation for external use containing original substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ in the patient A, 58 years, with burn. The burn of I, II degree of two fingers (middle finger - 1.5 cm ² and ring finger - 1 cm ² ) of left hand in the area of nail bed with overheated sunflower oil was diagnosed in the patient. The trauma was civilian. Hyperemia, cutaneous edema and big continuous blisters with contents were observed. On the next day after application of the preparation for external use in a form of gel significant reduction of hyperemia and almost complete absence of edema and content of blisters were observed. The preparation was applied for 5 days. On the 5 ^th day disruption of thin layer of vulnerary surface epidermis was observed; under this surface new skin without cicatrix and inflammation formed. After 14 days complete regeneration of damaged surface was observed. Example 19. Study of the preparation for external use containing original substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ in the patient T., 56 years old, with burn. The burn of I, II degree of the backside of left forearm in the area of fist, 20 cm ² , with hot vapor was diagnosed in the patient. The trauma was civilian. Hyperemia, cutaneous edema and big and small blisters with contents were observed. The preparation was applied for 3 days. On the next day after application of the preparation for external use in a form of ointment significant reduction of hyperemia and almost complete absence of edema and content of blisters were observed. On the 5 ^th day disruption of thin layer of vulnerary surface epidermis was observed; under this surface new skin without cicatrix and inflammation formed. After 14 days complete regeneration of damaged surface was observed.

Example 20. Study of the preparation for external use containing original substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ in the child S., 6 years old. Hyperemic scabrous plaque 1 cm ² that remained for 7 days was diagnosed on the right hip of the child in the area of contact with the elastic webbing. On the next day after application of the preparation for external use in a form of ointment the plaque disappeared and never appeared again. Example 21. Study of the preparation for external use containing original substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ . The patient G., 33 years old, with hyperemic plaque 1.5cm ² that remained for 10 days on the left side in the area of contact with the elastic webbing. On the next day after application of the preparation for external use in a form of spray the plaque disappeared and never appeared again.

Example 22. Study of the preparation for external use containing original substance with peptide structure Tyr-Pro-X where X - OH, NH ₂ , OCH ₃ , OC ₂ H ₅ . The patient K., 45 years old. On the left elbow there was a hyperemic plaque 3 cm ² that remained for long period of time. The patient addressed to medical institution on several occasions, but no results were observed. Two days after single application of the preparation in a form of plaster the plaque disappeared and never appeared again.