EP3914279A1

EP3914279A1 - Skin renewing and healing mixture of peptide components and its use

Info

Publication number: EP3914279A1
Application number: EP20709701.5A
Authority: EP
Inventors: Vladimir Kral; Pavel Martasek
Original assignee: Globetech Innovation SRO
Current assignee: Globetech Innovation SRO
Priority date: 2019-01-21
Filing date: 2020-01-20
Publication date: 2021-12-01
Also published as: JP7193662B2; IL284670A; CZ201934A3; WO2020152568A1; JP2022513418A; SG11202107397PA; CN113301910B; KR20210120026A; CZ308845B6; CN113301910A

Abstract

A skin renewing and healing mixture of peptide components significantly accelerates the healing of wounds. It can be used in particular for the treatment of chronic, flamed and complicated wounds, but also common injuries, post-operative wounds or burns. In healthy patients, the mixture reduces the time required for healing up to one third; in the case of difficult-to-heal wounds, the skin renewing and healing mixture of peptide components reduces the time required for healing up to 7 times. With chronic wounds, in particular decubitus and venous ulcers, the skin renewing and healing mixture of peptide components ensures the improvement of wound healing or even the complete healing of the wound as a result of the effect of the combination of the peptide components. Preferably, the skin renewing and healing mixture of peptide components is applied to wounds in a sterile protein and saccharide gel matrix preventing infection development thus allowing more efficacious wound healing. The skin renewing and healing mixture of peptide components can be applied in aesthetic medicine and cosmetics.

Description

Skin Renewing and Healing Mixture of Peptide Components and Its Use

Field of the Invention

Pharmaceutical and cosmetic preparations, formulations, and methods for topical or transdermal administration and treatment of wounds and/or for wound healing improvement. Skin therapy and treatment where the active substance are peptides and other biological components. Healing of wounds, including wounds difficult to heal, aesthetic medicine, cosmetics.

State of the Art

Under standard circumstances, the process of acute wound healing is divided into three (3) phases. The initial inflammatory phase followed by powerful remodelling and proliferation of tissues (proliferation phase) is further followed by the“mature phase” with re-epithelization, skin angiogenesis and the wound closure. Re-epithelization includes migration and proliferation of epithelial tissues, above all keratinocytes. Angiogenesis is the formation of new blood vessels from the already existing ducts that is regulated by a panoply of soluble cytokines, including growth factor polypeptides, as well as interactions of cellular cells and cellular matrices. Chronic wounds have a different treatment profile compared to standard acute wounds as they usually stay in the inflamed condition for a longer period. Non-healing wounds are most often found in patients with diabetes, venous stasis and in immobilised patients. Considering the aforementioned facts, it would be desirable to provide new biomolecules that safely and effectively facilitate the healing mechanism of epithelial and vascular wounds accompanying acute as well as chronic situations related to wound healing.

Chronic wounds have a different treatment profile compared to standard acute wounds as they usually stay in the inflamed condition for a longer period. Difficult-to-heal wounds are most often found in patients with diabetes, venous thrombosis and in immobilised patients. Considering the aforementioned facts, it would be desirable to provide new biomolecules that would specifically, safely and effectively facilitate the healing mechanism of epithelial and vascular wounds accompanying acute as well as chronic situations related to wound healing. Persons with diabetes have a very low skin regeneration capacity. Skin-related complications of diabetes result in the skin that is dry, prone to cracking and slow healing. Persons with diabetes often suffer from slow and insufficient skin healing. Pain in legs and ulcers are the main cause of lower limb amputation in diabetics and currently, approximately 10% of patients with diabetes require amputation during the course of their lives. The primary reason for amputation is infections related to the formation of ulcers and damage to skin that is incurable at the present time. The average risk of extremity amputation in persons with diabetes is 15 times higher compared to that in persons who do not have diabetes.

The currently employed treatment of chronic wounds is not able to reach efficacious therapeutic results. Most of these pharmaceuticals focus on angiogenesis but diabetes often encompasses endothelial dysfunction. Regenerative healing features quick and efficacious re-epithelization. There is a need for new, more efficacious treatment of chronic wounds in diabetic patients. Insufficient migration of epithelial cells is the characteristic feature of non-healing wounds and diabetes often involves endothelial dysfunction. Therefore, focus on re-epithelization that involves in particular keratocytes may improve the therapeutic results of the currently employed treatment.

The currently employed treatment of non-healing wounds and varicose ulcers is unsatisfactory. No optimal method of treatment of skin defects has been discovered so far.

A new method of treatment of venous ulcers utilizing the patient’s autologous cells has been developed. However, such a treatment is demanding and protracted. The currently employed treatment of chronic wounds is not able to reach efficacious therapeutic results. Most of these pharmaceuticals focus on angiogenesis but diabetes often encompasses endothelial dysfunction.

Recently, various pharmaceutical products supporting healing of wounds have been developed. Examples of these include Beclapermin, a genetically modified recombinant PDGF by Johnson & Johnson, or a pharmaceutical composition for regeneration and recovery of mammalian tissues comprising PDGF and Dexamethasone (EP 0575484). The document US 5 981 606 discloses a preparation for healing of wounds comprising TGF -beta and the documents US 6 800 286 and US 5 155 214 disclose preparations for healing of wounds comprising FGF. All the aforementioned pharmaceutical products are growth factors, cytokines or chemokines, collagen or hyaluronic acid. The disadvantage of these pharmaceutical products are adverse effects as they are not single cell type specific.

Peptides have control effects, which means that they are able to affect living cell behaviour. For example, the GHK (glycyl-L-histidyl-L-lysine) peptide, as a natural modulator of several cellular pathways for skin regeneration, is present in human plasma, saliva and urine, but its concentration decreases with age. GHK facilitates the intake of trace elements necessary for the regeneration of skin, such as copper. The peptide acts in complex with Cu²⁺, facilitates the healing of wounds and skin and participates in enzymatic processes ( Gorouhi F, Mai bach HI Role of topical peptides in preventing or treating aged skin. Int J Cosmet Sci. 2009; 31: 327-345). GHK stimulates both synthesis and decomposition of collagen and glycosaminoglycans and modulates the activity of metalloproteases and their inhibitors. It stimulates collagen, dermatan sulphate, chondroitin sulphate, low-molecular proteoglycan or decorin. It also facilitates the regeneration of the replication vitality of fibroblasts after radiotherapy. The molecule attracts immune and endothelial cells to the site of the wound.

Among other peptides used for therapy or cosmetic treatment of complexion and/or skin are signal peptides Syn®-Coll (Palmitoyl Tripeptide-5) with the Pal-Lys-Val-Lys-OH structure that bind directly to the receptor ( Brzoska T, Bohm M, Liigering A, Loser K, Luger TA. Terminal signal: anti-inflammatory effects of melanocyte-stimulating hormone related peptides beyond the pharmacophore. Adv Exp Med Biol. 2010; 681: 107-116). In addition, Decorinyl™ (tetrapeptide; Lupo MB, Cole AL. Cosmeceutical peptides. Dermatol Ther. 2007; 20: 343-349) stimulating fibrioblasts to produce collagen and the proliferation of elastin, glycosaminoglycan, proteoglycan and fibronectin. Peptides inhibit the following neurotransmitters: Argireline® with the following structure N-Acetyl-L-a-glutamyl-L-a-glutamyl-L-methionyl-L-glutaminyl- L-arginyl-L-argininamide, Vialox® with the following structure H-Gly-Pro-Arg-Pro-Ala- NH2, and Syn®-ake ( Lupo MP, Cole AL Cosmeceutical peptides. Dermatol Ther. 2007; 20: 343- 349) reduce the contraction of face muscles and consequently the formation wrinkles by increasing the minimal threshold for muscular activity.

Peptides inhibiting the following enzymes: Glycine soya protein (Preregen®; Siidel KM, Venzke K, Mielke H, Breitenbach U, Mundt C, Jaspers S, et al. Novel aspects of intrinsic and extrinsic aging of human skin: beneficial effects of soy extract. Photochem Photobiol. 2005; 81: 581-587. Andre-Frei V, Perrier E, Augustin C, Damour O, Bordat P, Schumann K, et al. A comparison of biological activities of a new soya biopeptide studied in an in vitro skin equivalent model and human volunteers. Int J Cosmet Sci. 1999; 21: 299-311) and sericin (Lupo ALP, Cole AL. Cosmeceutical peptides. Dermatol Ther. 2007; 20: 343-349) directly or indirectly inhibit the enzyme involved in the process ageing to the receptor (Brzoska T, Bohm M, Liigering A, Loser K, Luger TA. Terminal signal: anti inflammatory effects of a±-melanocyte-stimulating hormone related peptides beyond the pharmacophore. Adv Exp Med Biol. 2010; 681: 107-116). They stimulate fibrioblasts to produce collagen, the proliferation of elastin, glycosaminoglycans, proteoglycan and fibronectin. Alaptid, 8-methyl-6,9-diazaspiro[4.5]decane-7,10-dione, or Cyclo(L-alanyl-l -amino- 1- cyclopentancarbonyl), C9H14N2O2, Mr 182.2 g/mol, CAS: 90058-29-0, belongs to the group of substances that inhibit the release of the hormone L-prolyl-L-leucylglycinamide that stimulates melanocytes (MIF). The use of the MIF hormone itself as a therapeutic agent is limited by its easy enzymatic hydrolysis. A series of the MIF spiroderivatives has been prepared in particular with a view to eliminating this disadvantage {Kasafirek E. et al. Cs. pat. 231 227, 1986; US pat. 5,318,973, 1994; Cs. pat. 260 899, 1989). Alaptid has been selected as the best analog both from the point of view of enzymatic stability and considering its pharmacodynamic profile. In addition to other effects, Alaptid has been proven as a substance with a significant curative effect in experimental animal models {Kasafirek E. et al. Cs. pat. 276270, 1992).

Alaptid is likely to have a negative impact on the inhibition of the release of the melanocyte stimulating hormone thus increasing the concentration of melanocytes in epidermis. Melanocytes influence the formation and function of keratocytes via organelles known as melanosomes to a great extent. {McGrath J.A., EadyRA., Pope F.M. Rook's textbook of dermatology, 7th ed. Blackwell Publishing, 2004, pp.3-7; James W., Berger T. Elston D. Andrews' diseases of the skin: Clinical dermatology, 10th ed. Saunders, 2005, pp. 5-6). Keratinocytes migrate from stratum basale via stratum spinosum and stratum granulosum into stratum corneum where they facilitate the recovery of eidermis. {Watt F.M. The epidermal keratinocyte. BioEssays 1988, 8, 163- 167).

Alaptid is a compound (log P equals -0.67) accompanied by a low value of solubility in water and other protic solvents. This fact results in certain disadvantages such as a partial separation of Alaptid during processing in the form of a white cover on the treated wounds. A lower concentration of Alaptid dissolved in formulation can result in a lower quantity of the active substance absorbed via stratum corneum. Results of the use of Alaptid nanoparticles have been published recently, e.g. PV 2011-232. The limited solubility of Alaptid in the physiological environment does not allow it curative potential to be utilized to the full extent. With the preparation of liquid forms containing the active substance, the low solubility of Alaptid in a hydrophilic medium represents a significant disadvantage that may lead up to partial separation of Alaptid in the site subject to healing. The low solubility of Alaptid may also be the cause of its reduced absorption in the lower layer of epidermis.

The effects of Alaptid are known from many publications, such as, for example, from the document Sklendf, Zbynek, et al. “Formulation and release of alaptide from cellulose-based hydrogels.”, Acta Veterinaria Brno 81.3 (2013): 301-306 describing the release of Alaptid from cellulosic hydrogels.

The use of amino acids and peptides for the regeneration of tissues is well known. For the purposes of sports-related regeneration, amino acids are combined with peptides to regenerate muscles, for example, for oral administration. Amino acids and peptides are also comprised in a number of cosmetic formulations preventing skin ageing. Such cosmetic products reduce complexion/skin dryness, protect and hydrate skin, improve its flexibility and youthful appearance and fill in superficial as well as deeper wrinkles. The properties of amino acids and peptides facilitating the regeneration of tissues are known, but in relation to complexion/skin they are accentuated predominantly by cosmetic and aesthetic medicine.

The document EP 1640041 discloses a cosmetic composition for topical treatment of especially wrinkled skin or skin exposed to intense photodamage. The cosmetic composition includes an agent stimulating collagen synthesis being tri- up to hexapeptides and an agent increasing interaction between the extracellular matrix and fibroblasts. The cosmetic composition may also comprise other cosmetically active substances, such as extracts, UV filters, moisturizing substances or amino acids.

The document US20050209131 discloses a cosmetic composition comprising amino acids and peptide complex with copper. This composition is employed in cosmetic preparations such as body lotion or cosmetic pre-moistened wipes preventing skin from drying and ensuring its protection.

The document W003030926 discloses an aqueous solution of a peptide complex with copper comprising at least one amino acid with allegedly curative effects. However, the aqueous solutions are not tested in the aforementioned document, which fact raises suspicions as to the efficacy of such solutions.

The document US20130108700 discloses a composition of a gelatine matrix comprising a peptide and an amino acid. Such a formulation is applied by injection in the proximity of fresh wounds to prevent scar formation. Testing of the composition was performed after gynaecological surgeries where the patients were applied the composition into mucous membranes surrounding the site of incision. This procedure really reduced the scarring of the mucous membranes in the tested patients and increased the patients’ chances of successful pregnancy.

The document W02005042048 discloses a similar composition as the document US20130108700. However, in this case the composition is employed to heal bones, cartilages, ligaments, and tendons. The composition is also applied by injection.

Description of the Invention

The invention discloses a skin renewing and healing mixture of peptide components that shows surprising efficacy in quick healing of wounds, including difficult-to-heal diabetic wounds and that is based on a combination of peptide with at least one of the following components: a peptide (2-40 amino acids); an amino acid (arginine and/or cysteine in L-, D- or racemic form). It has been proven that a mixture of the aforementioned components has up to 100% higher in vivo efficacy in healing wounds in animal models and humans than the components applied individually.

The amino acids in a mixture of topically curative peptide components act as donors of nitrous oxide (NO) or sulfane (H2S).

The skin renewing and healing mixture of peptide components comprises at least one peptide in concentration ranging from 0.001 to 50 mg/ml or g of the matrix or formulation. It has been proven based on in vivo tests that any increase in the concentration of peptide(s) above 0.001 mg/ml or g of the matrix results in therapeutic effect while an increase in the concentration of peptide(s) above 50 mg/ml or g of the matrix results in no desired therapeutic effect.

The pH value of the formulation ranges between 3.0 and 9.0. Preferably, the formulation has ionic strength at least 20 mM, more preferably 50 mM up to 100 mM, and the most preferably at least 120 mM prior to the formulation application.

The skin renewing and healing mixture of peptide components can be dissolved in one cosmetically or pharmaceutically acceptable solvent or in a mixture of cosmetically or pharmaceutically acceptable solvents such as water, ethanol, propanol, isopropanol, propylene glycol, butylene glycol, dipropylene glycol, ethoxylate or propoxylated diglycols, cyclic polyols. The skin renewing and healing mixture of peptide components can be preferably dissolved with a pharmaceutical or cosmetic vector, such as liposomes, or the mixture of topically curative peptide components can be adsorbed onto powder organic polymers or powder minerals - such as talc or bentonites. Generally, the mixture of topically curative peptide components can be fixed in any cosmetically or pharmaceutically acceptable vector. The skin renewing and healing mixture of peptide components can be used in combination with other active substances, cosmetically or pharmaceutically active ones. The final formulation is then used either for cosmetic purposes focused on care for complexion/skin, or as a medicament. The formulation has rejuvenating and revitalizing properties. The skin renewing and healing mixture of peptide components may comprise peptides with other groups, such as signal or protective ones (tBoc, tags). The peptides may also be associated with nanoparticles and/or liposomes, such as those carrying other active substance. Peptides may also be modified by glycosylation, pegylation, acetylation, methylation, ubiquitination, hydroxylation, palmitoylation, phosphorylation or otherwise, provided that such modification does not affect the curative properties of the peptide.

Surprisingly, it has been discovered that small peptides (fewer than 12 amino acids in sequence) are more efficacious in terms of wound healing improvement than full-length natural proteins. In addition, it has been discovered that such enhanced efficacy is not shared by all fragments of the full-length peptides. Surprisingly, the inventors have proven that the peptides according to the present invention are more efficacious in healing wounds than the full-length protein and that such increased efficacy is not shared by all fragments of the full-length healing peptides and proteins used for natural healing of wounds. Moreover, a combination of peptides with amino acids or other peptides has shown higher efficacy in healing than an individual peptide employed for the treatment.

In addition, it has been discovered that the presence of Cu or Zn in the peptide metalocomplex can increase the efficacy by 5 to 10 more percent compared to treatment by peptides without the presence of metal; the mixture of topically curative peptide components preferably includes both a basic peptide and its metalocomplex. The curative efficacy of such a preparation is increased by 8 to 12% compared to that of preparations comprising either peptide or peptide metalocomplex. The mixture of topically curative peptides preferably includes the Cu-GHK peptide metalocomplex, where Cu-GHK is a natural, autologous substance, the quantity of which, however, decreases with age. For this reason, the treatment of injuries and chronic wounds is more difficult at a higher age. The skin renewing and healing mixtures of peptide components that comprise peptide metalocomplex(es) show enhanced efficacy in particular in the case of non-healing wounds.

The metalocomplexes are prepared in the laboratory environment by a simple synthesis where the peptide is dissolved or suspended in anhydrous methanol (MeOH) or acetonitrile, a concentrated solution of the metal, preferably Cu²⁺ or Zn²⁺, is added to the mixture along with two base equivalents, preferably sodium methoxide (MeONa) or another strong base. The mixture is stirred at ambient temperature for a period of 40 to 120 minutes. Then the respective peptide metalocomplex is extracted and dried. The dried peptide metalocomplex is used in a number of embodiments of the skin renewing and healing mixture of peptide components.

The skin renewing and healing mixture of peptide components stimulates the proliferative growth of fibroblasts and epithelial cells, such as keratinocytes, has positive effects on the entire wound healing process in laboratory animals as well as human volunteers. During the testing phase, it was also discovered that the skin renewing and healing mixture of peptide components efficaciously fills in wrinkles and rejuvenates complexion, lightens dark spots and birth marks, and prevents a formation of scars after cosmetic surgeries, such as a removal of freckles and birth marks. The skin renewing and healing mixture of peptide components has also been used for the treatment of burns, after surgeries and microsurgeries, with common injuries or for larger regions of injured skin, such as grazed sites.

During the research, it was also proven that the diketopiperazine spirocyclic peptide, such as Alaptid in the aforementioned combinations, has properties that considerably improve the growth of dermal cells in concentrations that also have antimicrobial effects. This enormous advantage can be utilized in particular in difficult-to-heal, flamed or diabetic wounds where sepsis and infection are major obstacles preventing the wound from healing.

In addition, it has been proven that the skin renewing and healing mixture of peptide components also influences the function of metalloproteases that play a role in healing of both acute and chronic wounds due to the control of the degradation and deposition of intercellular substance in the wound. The skin renewing and healing mixture of peptide components increases the level of metalloproteases in the site of the wound, by which the degradation of intercellular substance in the wound is accelerated and healing of the wound enabled.

For the skin renewing and healing mixture of peptide components, 2 main groups of curative proteins, namely diketopiperazine spirocyclic peptides and linear peptides, were used. Both the groups can also be divided into natural and synthetic. The natural peptides, such as the GHK peptide (Gly-His-Lys) or DAHK peptide (Asp-Ala-His-Lys) and other, play a vital role in skin regeneration. They influence the proliferation of skin cells, their regeneration and longevity. It has been proven that a combination of natural peptides with synthetic ones is preferable. As an example of the synthetic proteins, the CAR protein or diketopiperazine peptides (e.g. Alaptid) can be taken. A combination of diketopiperazine peptides with the GHK peptide/Cu- GHK peptide in the ratio of 1 : 1 has been proven as beneficial to a great extent. The performance of the GHK peptide/Cu-GHK peptide mixture in combination with arginine has been proven and seems to preferable to the same extent and the treatment with such a mixture takes longer by approximately 1/10 compared to treatment with a mixture of diketopiperazine + GHK/Cu-GHK. A mixture of at least two types of diketopiperazine peptides is also preferable for the treatment. In the case of difficult-to-heal wounds a combination of two diketopiperazines is by up to 70% more efficacious than the application of one diketopiperazine applied individually. Also, a combination of the GHK protein / Cu-GHK protein with arginine in the presence of a protein, preferably collagen, fibrinogen or elastin, has proven to be beneficial. In the case of difficult-to-heal wounds, such as diabetic ones, its is preferable to add to the mixtures of peptides an amino acid, such as arginine, as a donor of NO, and/or the amino acid cysteine, as a donor of TkS. Addition of the amino acids results in efficacy improvement.

The diketopiperazine peptides used for the skin renewing and healing mixture of peptide components can be described by the following general formulas:

General structures:

Examples of specific chemical substances:

(S)-isomer:

General structures:

Examples of specific chemical substances:

For the skin renewing and healing mixture of peptide components also the derivatives of the diketopiperazine proteins were used, such as:

Alaptid with the following structural chemical name: 8-methyl-6,9-diazaspiro[4.5]decane- 7,10-dione

and structural formula:

Alaptid-CD complex; 6-methyl-MeAlaptid; 6,9-di-MeAlaptid, 6-EtAlaptid; 6-hexylAlaptid; 9-ethylAlaptid; 6-Pal alaptide 6-palmitoyl 8-methyl-6,9-diazaspiro[4.5]decane-7,10-dione; 8-methyl-6,9-diazaspiro[5.5]undecane-7,10-dione;

For the skin renewing and healing mixture of peptide components, also the following linear peptides with the sequence of amino acids or used name were employed:

a) Cu-GHK

b) DAHK

c) Carnosine AH d) PLG

e) TFA-VWV-OH

f) KDI-Citrulline

g) Pal-GHK

h) Pal-GQPR

i) KRFK

j) Pal-KVK-bisTFA

k) Pal-KTTKS-OH

l) Pal-GQPR

m) Pal-VGVAPG

n) GEKG

o) PKEK

P) FVAPFP

q) Mn-GHK

r) GPRPA

s) YAGFL

t) AP-Dab-NHBn-2 -Acetate

U) Exendin-4 HGEGTF T SDL SKQMEEE A VRLFIEWLKN GGP S S GAPPP S

V) YV SPGMKNVNWW SHWWHATD

w) X-NH-GPAG-CO-Y (X = H, Boc, Fmoc, Me, Et; Y = OH, OMe, OEt, ONa, OK, ONH4),

X) VYYVGRK

y) c [ W CKPKPKPRCH-NH2 ] )

z) QHREDGS

aa) YGRRRRRQRRRP

bb) H-LEPPPLKP AD ALGV -OH

cc) AG30 ML SLIFLHRLK SMRKRLDRKLRLWHRKN YP

dd) AG30/5C MLKLIFLHRLKRMRKRLKRKLRLWHRKRYK

ee) AH90 AT AWDF GPHGLLPIRPIRIRPLCG

ff) CW49 APFRMGICTTN

gg) Cys-KR12 KRIVKRIKKWLR

hh) Esculentin- 1 a( 1 -21 ) GIF SKL AGKKIKNLLI S GLKG

ϋ) hBD-1 DHYNCVSSGGQCLYSACPIFTKIQGTCYRGKAKCCK

jj) hBD-2 GIGDPVTCLKSGAICHPVFCPRRYKQIGTCGLPGTKCCKKP kk) Hi statin- 1 D SHEKRHHGYRRKFHEKHHSHREFPF Y GD Y GSNYLYDN

11) Hi statin-2 RKFHEKHHSHREFPF Y GD Y GSNYLYDN

mm) Hi stain-3 DSHAKRHHGYKRKFHEKHHSHRGYRSNYLYDN

nn) IDR-1018 VRLIVAVRIWRR

oo) LL-37 LLGDFFRK SKEKIGKEFKRI V QRIKDFLRNL VPRTE S

PP) Pep 19-2.5 GCKK YRRFRWKFKGKF WF W G

qq) PLL-37 PLLGDFFRK SKEKIGKEFKRI V QRIKDFLRNL VPRTE S

rr) SHAP1 APKAMKLLKKLLKLQKKGI

ss) SR-0007 MLKLIFLHRLKRMRKRLKRK

tt) SR-0379 MLKLIFLHRLKRMRKRLkRK

uu) Temporin A FLPLIGRVL S GIL

vv) Temporin B LLPI V GNLLK SLL

The skin renewing and healing mixture of peptide components may preferably be in the form of solution, suspension, emulsion, ointment, balm, tincture, elixir, patch, bandage, dressing material, alginate dressing or compress, topical solution, infusion or surgical wash. The matrix of the aforementioned forms preferably includes proteins.

The skin renewing and healing mixture of peptide components is preferably applied onto wounds in formulation with a gel matrix. The gel matrix comprises a protein and saccharide components in the ratio of 1 : 1 by weight and an aqueous buffer solution containing Ca²⁺, Mg²⁺, and Zn²⁺, preferably in the form of calcium acetate, zinc acetate or magnesium acetate. The pH value of the gel preferably ranges between 3 and 9. Thanks to the presence of cations, the gel is formed either by complexation with collagen and/or by cation-controlled gelation. The protein component is preferably collagen, split collagen, fibrinogen, split fibrinogen, elastin, split elastin, gelatine, hydrolysed gelatine, BSA, etc. The saccharide component is preferably hyaluronic acid, pectin, alginate, carrageenan, alginic acid, oxidized and non- oxidized forms of cellulose, cellulose derivatives - for example carboxymethyl cellulose and its salts, other carboxylated saccharides, chitosan, sorbitan caprylate, flaxseed gum, pectate and other oligo- and polysaccharides, etc. The matrix may also be formed by synthetic polymers, such as polyacryl amides, poly(lactic-co-glycolic acid) (PLGA) and polylactic acid (PLA).

The gel matrix provides skin hydration, nourishment and toning, is preferable for use in combination with the skin renewing and healing mixture of peptide components for the treatment of chronic wounds, decubitus ulcers, diabetic complications, and treatment following laser and cosmetic surgeries. In addition, in combinations with the skin renewing and healing mixture of peptide components it smooths wrinkles, decelerates the formation of new ones and efficaciously decelerates the process of complexion ageing, stimulates skin cells to produce collagen and elastin, brightens and equalizes the colour of the complexion. The gel applied to wounds in formulation with the skin renewing and healing mixture of peptide components is preferably sterile.

The matrix may also be oil in water or water in oil, and the peptide and/or amino acid and/or protein is preferably present in the fat phase. The fat phase preferably comprises also phosphatidylcholine or other lecithin. Lecithin is preferably present in the composition in the form of liposomes or micelles or other structures containing nitrous oxide, peptides or both. Preferably, the matrix for the skin renewing and healing mixture of peptide components is in the form of gel, cream, lotion, ointment, solution, solid“stick”, etc. that can be applied or sprayed onto the skin, e.g. injured.

The prepared skin renewing and healing mixture of peptide components attains high efficacy in accelerated wound healing. This result is based on shortened peptide chains used for healing and also by adding donors of NO and ELS. The skin renewing and healing mixture of peptide components reduces time required for healing in healthy patients nearly to one third compared to standard treatment (from 8 weeks to 3 weeks) and in ill patients with chronic dermal defects, the time required for healing is reduced up to 7 times (from 20 weeks to 3 weeks); in some patients with chronically inflamed wounds the skin renewing and healing mixture of peptide components allows treatment in otherwise unmaintainable cases intended for amputation. With the use of preparations comprising a mixture of peptide components - peptide + Arg + Cys as per Example 1, the wounds and/or dermal defects healed with such formulations were healed in murine models on average in 3 weeks, in healthy patients in 3 weeks, and in diabetics, healing was attained in 3 to 7 weeks. With preparations based on peptide + Arg/Cys as per Examples 2 and 3, or with preparations based on peptide + peptide + Arg + Cys as per Examples 5, 6, and 7, the wounds were healed in murine models in 4 weeks, in healthy patients in 4 weeks, and in the case of difficult-to-heal wounds, healing occurred between 4 and 9 weeks. With preparations peptide + peptide as per Example 4, the wounds in murine models were healed on average in 5 weeks, in healthy patients in 5 weeks and difficult-to-heal wounds healing took 5 to 12 weeks. With preparations as per Example 8A, i.e. control groups with the application of a peptide itself in a gel matrix (Alaptid was used as a model), the wounds in murine models were healed on average in 7 weeks, in healthy patients in 7 weeks and difficult-to-heal wounds healing took 7 to 20 weeks. With preparations as per Example 8B, i.e. control groups with the application of a gel matrix itself, the wounds in murine models were healed on average in 8 weeks, in healthy patients in 8 weeks and the healing of difficult-to-heal wounds took 8 to 20 weeks; however, only one patient was healed completely. With preparations as per Example 8C, i.e. control groups with no treatment, the wounds in murine models were healed on average in 8 weeks, in healthy patients in 8 weeks and the healing of difficult-to-heal wounds was completed only in one patient out of five, namely in 31 weeks with red spots remaining present in all tested subjects.

Summary of presented drawings

Fig. 1 : Comparison of the selected preparations used in murine models

Fig. 2: A histological image, day 21 of experimental wound healing in a mouse by the preparation mentioned in Example IB, enlarged 40 times

Fig. 3: A histological image, day 28 of experimental wound healing in a mouse by the preparation mentioned in Example 2, enlarged 40 times

Fig. 4: A histological image, day 28 of experimental wound healing in a mouse by the preparation mentioned in Example 7, enlarged 40 times

Fig. 5: A histological image, day 50 of experimental wound healing in a mouse by the preparation mentioned in Example 8B, enlarged 40 times

Fig. 6: Comparison of the selected preparations used in selected patients

Fig. 7: Table of wound healing with the use of the selected preparations

Fig. 8: Chart of wound healing with the use of the selected preparations

Examples of Invention Execution

Example 1

Preparation of the peptide metalocomplex

Cu-GHK

1.5 g of GHK was suspended at ambient temperature in 50 ml of 100% methanol, while stirred continuously 0.25 g of cupric acetate (CuOAc) was added in the form of 5 ml of aqueous solution and 5 ml of MeONa (1M). The mixture was stirred at ambient temperature at the speed of 130 rpm for 120 minutes. The peptide metalocomplex was then extracted and dried.

Production of the preparation - peptide + Arg + Cvs

1A 1% Cu-GHK vaseline

1 g of powder GHK peptide (Gly-His-Lys), 0.3 g of powder L-arginine, and 0.1 g of powder L- cysteine was mixed in 98.6 g of medicinal vaseline at 3,000 rpm. A preparation with 1% content of the Cu-GHK peptide was prepared.

IB 1% Cu-GHK matrix gel

1 g of powder Cu-GHK peptide (Gly-His-Lys), 0.3 g of powder L-arginine, and 0.1 g of powder L-cysteine was mixed in the gel matrix consisting of 7 g of collagen, 7 g of pectin, and 86 g of phosphate buffer at pH 7 with 5% calcium acetate. The ingredients of the gel matrix were mixed together and stirred for 5 minutes using a magnetic stirrer at 300 rpm and then for 25 more minutes at 130 rpm. Then the Cu-GHK peptide and L-Arg were added to the gel matrix and the mixture was stirred at 2,000 rpm for 5 more minutes.

1C 1.2% GHK vaseline

1.2 g of powder GHK peptide (Gly-His-Lys), 0.3 g of powder L-arginine, and 0.1 g of powder L-cysteine was mixed in 98.6 g of medicinal vaseline at 3,000 rpm. A preparation with 1% content of the GHK peptide was prepared.

ID 1% DAHK (1 : 0.3 : 0.1) vaseline

1 g of powder DAHK peptide (Asp-Ala-His-Lys), 0.35 g of powder L-arginine, and 0.1 g of powder L-cysteine was mixed in 98.6 g of medicinal vaseline at 3,000 rpm. A preparation with 1% content of the DAHK peptide was prepared. IE 0.5% DAHK matrix ointment

0.5 g of powder DAHK peptide (Asp-Ala-His-Lys), 0.15 g of powder D-arginine, and 0.08 g of powder D-cysteine was mixed in the matrix consisting of:

liquid paraffin 8.0 g, hard paraffin 12.0 g, stearyl alcohol 2.0 g, propylene glycol 5.5 g, Slovasol 2430 (1 -(2 -methoxyisopropoxy)-2 -propanol) 3.0 g, Carbomera O.5 g, triethanol amine 0.6 g, methylparaben 0.2 g, propylparaben 0.05 g, and purified water 67.65 g.

The mixture was stirred at 5,000 revolutions for 2 hours.

All components were homogenized into emulsion with the content of 0.5% of DAHK peptide.

IF 1% Alaptid vaseline

1 g of powder Alaptid, 0.4 g of powder L-arginine, and 0.1 g of powder L-cysteine was mixed in 98.6 g of medicinal vaseline at 3,000 rpm. A preparation with 1% content of Alaptid was prepared.

1G 5% Alaptid matrix emulsion

5 g of powder Alaptid was mixed in 4 g of refined coconut oil, emulsifier was added, and the mixture was mixed with 82 g of water. The emulsifier consisted of 9 g of 9% tridecyl alcohol ethoxy late.

In the presence of the emulsifier, the two immiscible liquids were mixed. The acquired poly disperse emulsion was mixed with 2 g of powder racemic arginine and 0.5 g of powder racemic cysteine and was additionally homogenized in a homogenizer at ambient temperature at 3,000 rpm. Homogenization improved the stability of the prepared emulsion. Emulsion with 5% content of Alaptid was prepared.

1H 2% Exendin-4 vaseline

2 g of powder Exendin-4, 0.3 g of powder L-arginine, and 0.1 g of powder L-cysteine was mixed in 98.6 g of medicinal vaseline at 3,000 rpm. A preparation with 2% content of the Exendin-4 was prepared.

II 1% 3-methyl-l,4-diazospiro[5.6]dodecane-2,5-dione vaseline

1 g of powder 3-methyl-l,4-diazospiro[5.6]dodecane-2,5-dione, 0.3 g of powder L-arginine, and 0.1 g of powder L-cysteine was mixed in 98.6 g of medicinal vaseline at 3,000 rpm. A preparation with 1% content of 3-methyl-l,4-diazospiro[5.6]dodecane-2,5-dione. 1 J 1% GEKG vaseline

1 g of powder GEKG, 0.5 g of powder L-arginine, and 0.2 g of powder L-cysteine was mixed in 98.6 g of medicinal vaseline at 3,000 rpm. A preparation with 1% content of GEKG was prepared.

Example 2

Production of the preparation - peptide + Arg

1% Zn-GHK matrix gel

1 g of powder Zn-GHK peptide (Gly-His-Lys) and 0.2 g of powder D-arginine was mixed in the gel matrix consisting of 7 g of gelatine, 7 g of carrageenan, and 86 g of phosphate buffer at pH 7 with 5% zinc acetate. The ingredients of the gel matrix were mixed together and stirred for 5 minutes using a magnetic stirrer at 300 rpm and then for 25 more minutes at 130 rpm. Then the Zn-GHK peptide and L-Arg were added to the gel matrix and the mixture was stirred at 2,000 rpm for 5 more minutes.

Example 3

Production of the preparation - peptide + Cys

1% DAHK matrix gel

1 g of powder DAHK peptide and 0.2 g of powder L-arginine was mixed in the gel matrix consisting of 2 g of gelatine, 1 g of chitosan, 1 g of hyaluronic acid and 3 g of collagen, 7 g of carrageenan, and 86 g of phosphate buffer at pH 7 with 5% magnesium acetate. The ingredients of the gel matrix were mixed together and stirred for 5 minutes using a magnetic stirrer at 300 rpm and then for 25 more minutes at 130 rpm. Then the DAHK peptide and L-Arg were added to the gel matrix and the mixture was stirred at 2,000 rpm for 5 more minutes.

Example 4

Production of the preparation - peptide + peptide in solution

4A GHK + Alaptid solution

1.5 g of powder GHK protein and 1 g of powder Alaptid was stirred in 97.5 g of purified water. The mixture was stirred at 5,000 rpm for a period of 15 minutes. Before every use, the mixture was stirred at 3,000 rpm for 2 minutes. 4B GHK + DAHK solution

1 g of powder GHK and 1 g of powder DAHK was stirred in 98 g of purified water. The mixture was stirred at 4,500 rpm for 10 minutes. Before every use, the mixture was stirred at 3,000 rpm for 1 minute.

4C Alaptid + 4,6,7-triethyl-4,7-diazospiro [2.5] octane-5, 8-dione solution

1 g of powder Alaptid and 1.5 g of powder 4,6,7-triethyl-4,7-diazospiro[2.5]octane-5,8-dione was stirred in 97.5 g of purified water. The mixture was stirred at 5,000 rpm for 20 minutes. Before every use, the mixture was stirred at 3,000 rpm for 2 minutes.

Example 5

Production of the preparation - peptide + peptide + Arg matrix gel

1 g of powder DAHK peptide and 1 g of powder GHK peptide, and 0.2 g of powder L-arginine was mixed in the gel matrix consisting of 6 g of hydrolysed gelatine, 1 g of BSA, 5 g of carboxymethyl cellulose, 2 g of hyaluronic acid and 86 g of borate buffer at pH 6 with 5% calcium acetate. The ingredients of the gel matrix were mixed together and stirred for 5 minutes using a magnetic stirrer at 300 rpm and then for 25 more minutes at 130 rpm. Then the DAHK peptide, GHK peptide, and L-Arg were added to the gel matrix and the mixture was stirred at 3,000 rpm for 5 more minutes.

Example 6

Production of the preparation - peptide + peptide + Cys matrix gel

1 g of powder DAHK peptide and 1 g of powder Alaptid, and 0.2 g of powder L-cysteine was mixed in the gel matrix consisting of 3 g of split elastane, 1 g of hydrolysed gelatine, 3 g of BSA, 5 g of carboxymethyl cellulose, 1 g of carrageenan, 1 g of alginic acid, and 86 g of borate buffer at pH 7.5 with 1 % zinc acetate. The ingredients of the gel matrix were mixed together and stirred for 15 minutes using a magnetic stirrer at 400 rpm and then for 25 more minutes at 130 rpm. Then DAHK peptide, Alaptid, and L-Cys were added to the gel matrix and the mixture was stirred at 5,000 rpm for 2 more minutes.

Example 7

Production of the preparation - peptide + peptide + Arg + Cys matrix gel 1 g of powder GHK peptide and 1 g of powder Alaptid and 0.5 g of powder L-arginine, and 0.2 g of L-cysteine was mixed in the gel matrix consisting of 6 g of split collagen, 1 g of fibrinogen, 4 g of oxocellulose, 1 g of alginate, 1 g of pectate, 1 g of flaxseed gum, and 86 g of phosphate buffer at pH 7 with 1% zinc acetate. The ingredients of the gel matrix were mixed together and stirred for 15 minutes using a magnetic stirrer at 400 rpm and then for 25 more minutes at 130 rpm. Then DAHK peptide, Alaptid, and L-Cys were added to the gel matrix and the mixture was stirred at 5,000 rpm for 2 more minutes.

Example 8

8A Production of the preparation - peptide matrix gel

3 g of powder Alaptid were mixed in the gel matrix consisting of 4 g of collagen, 3 g of split collagen, 1 g of hyaluronic acid, 3 g of alginate, 3 g of carrageenan, and a 86 g of acetate buffer at pH 6.5 with 3% magnesium acetate. The ingredients of the gel matrix were mixed together and stirred for 15 minutes using a magnetic stirrer at 400 rpm and then for 25 more minutes at 130 rpm. Then Alaptid was added to the gel matrix and the mixture was stirred at 5,000 rpm for 5 more minutes.

8B Production of the preparation - matrix gel

The control gel matrix was prepared from 7 g of collagen, 7 g of pectin, and 86 g of phosphate buffer at pH 7 with 5% calcium acetate. The ingredients of the gel matrix were mixed together and stirred for 5 minutes using a magnetic stirrer at 300 rpm and then for 25 more minutes at 130 rpm. Then the gel matrix was stirred at 2,000 rpm for 1 more minute.

Example 9

Testing of individual preparation efficacy

To compare efficacy, preparations based on individual examples were produced in a standardized concentration by mixing with medicinal vaseline. This approach guaranteed the uniformity of the tested formulations. In addition, all produced formulations were tested. Figures 7 and 8 show the in vivo number of days required for complete healing of wounds in individual tested groups. The selected formulations representing a given group of preparations are shown. The results show that the effect of the formulation is independent of the used peptide but depends on the combination of peptide(s) and amino acids in a certain ratio. To compare the efficacy of the preparations, animal models of inbred laboratory mice were selected and were caused artificial wounds in abdominal cavity using a sterile scalpel. Each experimental subject was caused 2 wounds approximately 1 cm long and approximately 3 mm deep. Following tests in murine models, testing is healthy injured patients and in diabetic injured patients and/or patients with non-healing wounds was performed. In the case of the healthy patients, in particular wounds resulting from microsurgeries, in diabetics difficult-to-heal wounds, venous ulcers, decubitus ulcers and fistulas were concerned. The tested group of mice and patients always included 5 tested subjects. The wounds were applied 0.002 g of peptide on a daily basis, i.e. 0.2 g of 1% peptide formulation a day. Tests on mice were assessed every 24 hours, tests on patients were recorded on a daily basis by the patients themselves - a photograph with a short description of the wound was taken every day. The records were assessed by the attending physician later on.

It has been discovered that the best effect of accelerated wound healing was attained if the preparations as per Example 1, i.e peptide + Arg + Cys were used. In addition, it has been proven that the formulation in the gel matrix has synergistic effect compared to the formulation of a mixture of curative peptide component in a vaseline matrix. The formulation in the gel matrix provides for quicker healing of wounds. The wounds treated by the formulations as per Example 1 were healed in murine models on average in 3 weeks, in healthy patients in 3 weeks and difficult-to-heal wounds healing took 3 to 7 weeks. With preparations as per Examples 2, 3, and 7, the wounds in murine models were healed on average in 4 weeks, in healthy patients in 4 weeks and healing of difficult-to-heal wounds took 4 to 9 weeks. With preparations as per Example 4, the wounds in murine models were healed on average in 5 weeks, in healthy patients in 5 weeks and the healing of difficult-to-heal wounds took 5 to 12 weeks. With preparations as per Examples 5 and 6, the wounds in murine models were healed on average in 4 weeks, in healthy patients in 4 weeks and difficult-to-heal wounds healing took 4 to 8 weeks. With preparations as per Example 8A, i.e. control groups with the application of a peptide itself in a gel matrix (Alaptid was used as a model), the wounds in murine models were healed on average in 7 weeks, in healthy patients in 7 weeks and difficult-to-heal wounds healing took 7 to 20 weeks. With preparations as per Example 8B, i.e. control groups with the application of a gel matrix itself, the wounds in murine models were healed on average in 8 weeks, in healthy patients in 8 weeks and the healing of difficult-to-heal wounds took 8 to 20 weeks; however, only one patient was healed completely. With preparations as per Example 8C, i.e. control groups with no treatment, the wounds in murine models were healed on average in 8 weeks, in healthy patients in 8 weeks and the healing of difficult-to-heal wounds was completed only in one patient out of five, namely in 31 weeks with red spots remaining present in all tested subjects.

Example 10

10A Effects of the preparation as per Example IB, anti-ageing, wrinkle smoothing

A group of 35 women at the age of 30 to 60 was tested. Prior to testing, photos of the faces of all the women were taken by the camera Canon 5D mark III (full frame) with the lens Canon EF 100 2,8L USM. The photographs were taken with the shutter 11, ISO 100 with the face exposed to three studio luminaires with the output of 500 W. The group was divided into a control and tested subjects. The control part of the group was applied a placebo preparation, while the tested subjects were applied the preparation as per Example IB. The placebo was the gel matrix without added peptide and amino acids.

The women applied the preparation as per Example IB twice a day for a period of 6 weeks. After testing completion, the faces of the women were documented again, i.e. photos of their faces were taken by the macro lens camera. The photo of each woman before and after testing were compared by image analysis. On average, 70% smoothing of wrinkles was attained with twice-a-day application for 6 weeks, while in the control group the smoothing of wrinkles amounted to 8% probably due to hydration provided by the gel matrix.

10B Effects of the preparation as per Example 7, anti-ageing, wrinkle smoothing

A group of 35 women at the age of 28 to 59 was tested. Prior to testing, photos of the faces of all the women were taken by the camera Canon 5D mark III (full frame) with the lens Canon EF 100 2,8L USM. The photographs were taken with the shutter 11, ISO 100 with the face exposed to three studio luminaires with the output of 500 W. The women applied the preparation as per Example 7 twice a day for 6 weeks. After testing completion, the faces of the women were documented again, i.e. photos of their faces were taken by the macro lens camera. The photo of each woman before and after testing were compared. On average, 62% smoothing of wrinkles was attained with application twice a day for 6 weeks.

Applicability in Industry

Preparations for accelerated healing of wounds, burns, non-healing and diabetic wounds, aesthetic medicine and cosmetics.

Claims

1. A skin renewing and healing mixture of peptide components characterized by the fact, that it comprises at least two peptide components from the following group: a peptide consisting of 2 to 40 amino acids, arginine amino acid, cysteine amino acid, where at least one of the peptide components is a peptide consisting of 2 to 40 amino acids, and the peptide consisting of 2 to 40 amino acids is a linear peptide or a spirocyclic diketopiperazine peptide.

2. The skin renewing and healing mixture of peptide components according to claim 1 characterized by the fact, that it comprises the peptide consisting of 2 to 40 amino acids, arginine amino acid, and cysteine amino acid.

3. The skin renewing and healing mixture of peptide components according to claim 2 characterized by the fact, that it comprises a linear peptide, a spirocyclic diketopiperazine peptide, arginine amino acid, and cysteine amino acid.

4. The skin renewing and healing mixture of peptide components according to claim 1 characterized by the fact, that at least one peptide consisting of 2 to 40 amino acids is a metalocomplex peptide.

5. The skin renewing and healing mixture of peptide components according to claim 4 characterized by the fact, that the metalocomplex peptide is Cu-GHK.

6. The skin renewing and healing mixture of peptide components according to claim 1 characterized by the fact, that the linear peptide is selected from the group with the following sequence of amino acids: GHK, Cu-GHK, Zn-GHK, DAHK.

7. The skin renewing and healing mixture of peptide components according to claim 1 characterized by the fact, that it comprises a linear peptide and a spirocyclic diketopiperazine peptide.

8. The skin renewing and healing mixture of peptide components according to claim 1 characterized by the fact, that it comprises a mixture of linear peptides.

9. The skin renewing and healing mixture of peptide components according to claim 1 characterized by the fact, that it comprises amixture of spirocyclic diketopiperazine peptides.

10. The skin renewing and healing mixture of peptide components according to claim 1 characterized by the fact, that the spirocyclic diketopiperazine peptide has a general formula

R¹ = -CH₃_ -CH₂CH₃ -(CH₂)₂CH₃, -(CH₂)₇CH₃

R² = -H, -CH₃ -CH₂CH₃ -(CH₂)₇CH₃

R³ = -H, -CH₃ -CH₂CH₃ -(CH₂)₇CH₃ -(CH₂)₁₇CH₃

11. The skin renewing and healing mixture of peptide components according to claim 1 characterized by the fact, that the spirocyclic diketopiperazine peptide is Alaptid - 8-methyl-6,9-diazaspiro[4.5]decane-7,10-dione with the following structural formula

12. The skin renewing and healing mixture of peptide components according to claim 1 characterized by the fact, that the linear peptide is selected from the group with the following sequence of amino acids: a) Cu-GHK

b) DAHK

c) Ala-His

d) PLG e) TFA-VWV-OH

f) KDI-Citrulline

g) Pal-GHK

h) Pal-GQPR

i) KRFK

j) Pal-KVK-bisTFA

k) Pal-KTTKS-OH

l) Pal-GQPR

m) Pal-VGVAPG

n) GEKG

o) PKEK

p) FVAPFP

q) Mn-GHK

r) GPRPA

s) YAGFL

t) AP-Dab-NHBn-2-Acetate

u) HGEGTF T SDL SKQMEEE A VRLFIEWLKN GGP S S GAPPP S v) YV SPGMKNVNWW SHWWHATD

w) X-NH-GPAG-CO-Y (X = H, Boc, Fmoc, Me, Et; Y = OH, OMe, OEt, ONa, OK, ONH4),

x) VYYVGRK

y) c [ W CKPKPKPRCH-NH2 ] )

z) QHREDGS

aa) YGRRRRRQRRRP

bb) H-LEPPPLKP AD ALGV -OH

cc) ML SLIFLHRLK SMRKRLDRKLRLWHRKN YP

dd) MLKLIFLHRLKRMRKRLKRKLRLWHRKRYK

ee) AT AWDF GPHGLLPIRPIRIRPLCG

ff) APFRMGICTTN

gg) KRIVKRIKKWLR

hh) GIF SKL AGKKIKNLLI S GLKG

ii) DHYNCVSSGGQCLYSACPIFTKIQGTCYRGKAKCCK

jj ) GIGDP VTCLKSGAICHPVFCPRRYKQIGTCGLPGTKCCKKP kk) D SHEKRHHGYRRKFHEKHHSHREFPF Y GD Y GSNYLYDN 11) RKFHEKHHSHREFPFYGDYGSNYLYDN

mm) DSHAKRHHGYKRKFHEKHHSHRGYRSNYLYDN

nn) VRLI V A VRFWRR

oo) LLGDFFRK SKEKIGKEFKRI V QRIKDFLRNL VPRTE S

pp) GCKK YRRFRWKFKGKF WF W G

qq) PLLGDFFRK SKEKIGKEFKRI V QRIKDFLRNL VPRTE S

rr) APKAMKLLKKLLKLQKKGI

ss) MLKLIFLHRLKRMRKRLKRK

tt) MLKLIFLHRLKRMRKRLKRK

uu) FLPLIGRVL S GIL

vv) LLPI V GNLLK SLL

13. The skin renewing and healing mixture of peptide components according to claim 1 characterized by the fact, that it is included in a protein or fat or saccharide or synthetic polymeric matrix or a mixture thereof in the quantity ranging from 0.001 to 50 mg per gram of matrix.

14. The skin renewing and healing mixture of peptide components according to claim 13 characterized by the fact, that it is included in a protein or fat or saccharide matrix or a mixture thereof in the quantity ranging from 0.1 to 5 mg per gram of matrix.

15. The skin renewing and healing mixture of peptide components according to claim 13 characterized by the fact, that the protein matrix comprises collagen.

16. The skin renewing and healing mixture of peptide components according to claim 13 characterized by the fact, that the saccharide matrix comprises chitosan and/or hyaluronic acid.

17. The skin renewing and healing mixture of peptide components according to claim 13 characterized by the fact, that the fat matrix comprises lecithin.

18. The skin renewing and healing mixture of peptide components according to claim 13 characterized by the fact, that the synthetic polymeric matrix comprises polyacryl amide.

19. The skin renewing and healing mixture according to claim 1 for the use in the treatment of dermal defects and/or for wound healing and/or skin regeneration.