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(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2013/020527 Al 14 February 2013 (14.02.2013) P O P C T

(51) International Patent Classification: (74) Common Representative: UNIVERSITY OF VETER¬ A61K 9/06 (2006.01) A61K 47/32 (2006.01) INARY AND PHARMACEUTICAL SCIENCES A61K 9/14 (2006.01) A61K 47/38 (2006.01) BRNO FACULTY OF PHARMACY; University of A61K 47/10 (2006.01) A61K 9/00 (2006.01) Veterinary and Pharmaceutical Sciences Brno Faculty Of A61K 47/18 (2006.01) Pharmacy, Palackeho 1/3, CZ-61242 Brno (CZ). (21) International Application Number: (81) Designated States (unless otherwise indicated, for every PCT/CZ20 12/000073 kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, (22) Date: International Filing BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, 2 August 2012 (02.08.2012) DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (25) Filing Language: English HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, (26) Publication Language: English ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, (30) Priority Data: NO, NZ, OM, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, 201 1-495 11 August 201 1 ( 11.08.201 1) SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, 2012- 72 1 February 2012 (01.02.2012) TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, 2012-5 11 26 July 2012 (26.07.2012) ZW. (71) Applicant (for all designated States except US): UNIVER¬ (84) Designated States (unless otherwise indicated, for every SITY OF VETERINARY AND PHARMACEUTICAL kind of regional protection available): ARIPO (BW, GH, SCIENCES BRNO FACULTY OF PHARMACY GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, SZ, TZ, [CZ/CZ]; Palackeho 1/3, CZ-61242 Brno (CZ). UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, (72) Inventors; and EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, FT, LT, LU, LV, (71) Applicants : JAMPILEK, Josef [CZ/CZ]; Husova MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, 202/17, CZ-29301 Mlada Boleslav (CZ). OPATRILOVA, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, Radka [CZ/CZ]; Ukrajinska 25, CZ-62500 Brno - Bohu- ML, MR, NE, SN, TD, TG). nice (CZ). COUFALOVA, Lenka [CZ/CZ]; Sladkova Declarations under Rule 4.17: 439/11, CZ-37701 JindficMv Hradec (CZ). CERNIKOVA, Aneta [CZ/CZ]; CZ-5695 1 Morasice 152 (CZ). — of inventorship (Rule 4.17(iv)) DOHNAL, Jiff [CZ/CZ]; Detska 2506/42, CZ-10000 Published: Praha 10 - Strasnice (CZ). — with international search report (Art. 21(3))

o (54) Title: UTILIZATION OF ALAPTIDE AS TRANSDERMAL PENETRATION MODIFIER FN PHARMACEUTICAL COMPOSITIONS FOR HUMAN AND VETERINARY APPLICATIONS CONTAFNING ANTI-FNFLAMMATORY DRUGS AND/OR ANTIMICROBIAL CHEMO- © THERAPEUTICS (57) Abstract: The invention deals with the way of utilization of micronized, nanonized and/or surface-modified alaptide, which af o fects penetration of other pharmaceutically active compounds through the skin as a pharmaceutical adjuvant (excipient). These phar maceutical compositions composed of alaptide as the excipient, pharmaceutical active ingredients (non-steroidal anti-inflammatory drugs and/or /non- and/or and/or antimicrobial chemotherapeutics, i.e. antibacterials, o antimycotics, antivirotics) and other pharmaceutical excipients can be used for preparation of drug formulations, which can influence the drug level in the body in time and can be used for both local and systemic administration. TITLE:

Utilization of Alaptide as Transdermal Penetration Modifier in Pharmaceutical Compositions for Human and Veterinary Applications Containing Anti-inflammatory Drugs and/or Antimicrobial Chemotherapeutics.

TECHNICAL FIELD:

The invention deals with utilization of (¾-8-methyl-6,9-diazaspiro[4.5]decan-7,10-dione, known by the international non-proprietary name of "alaptide", as a pharmaceutical adjuvant (excipient) for modification of transdermal penetration of drugs in pharmaceutical formulations convenient for transdermal application, i.e. for preparation of human and/or veterinary drug formulations, which can influence the drug level in the body in time and can be used for both local and systemic administration of non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/ antimycobacterials, antimycotics, antivirotics).

STATE OF THE ART:

Background

The recent development in the field of pharmaceutical dosage forms results in the discovery of additional highly sophisticated drug delivery systems that allow maintaining a stationary level of the active substance in an organism. Transdermal therapeutic systems represent an excellent alternative to conventional pharmaceutical dosage forms. However, the application of transdermal drug delivery faces the problem of insufficient or no penetration of active pharmaceutical substances through the skin. To solve this critical issue some physical or chemical possibilities/approaches for overcoming the skin barrier were developed. The idea of percutaneous drug absorption appeared long ago; the Ebers papyrus mentioned it as early as the 16th century B.C. In 1975 Idson stated that the epidermic barrier was a limiting factor for percutaneous absorption and that once the drug passed through stratum rne m (SC) of the epidermis, its absorption was guaranteed. A significant search for chemical substances improving skin permeability has been the subject of extensive studies during the last decades. More than 350 different compounds are known as chemical penetration enhancers perturbing the SC barrier to facilitate drug delivery. The largest increase in the number of enhancers was noted in the 80s of the last century. At present it can be said that the active pool of such substances is rather stable. At the time when new chemical entities are being discovered with exponential speed (which is evident from the Chemical Abstract Service lists), the retention of the number of chemical penetration enhancers is quite surprising. The source of this anomaly is an apparent insufficient understanding of mechanistic principles determining the power of enhancers, low efficiency of experimental effect determination and the fact that the enhancers have not reached their full potential in transdermal or topical systems so far (Prausniiz, M.R. et al. Nature Rev. Drug Discov. 2004, 3, 115; Rabiskovd M. et al. Technology of Pharmaceutics, 3rd ed., Galen Prague, 2006; Idson, B. J. Pharm. Sci. 1975, 64, 901; Pfister, W.R. et al. Pharm. Tech. 1990, 14, 132; Finnin, B.C.; et al. J. Pharm. Sci. 1999, 88, 955; Karande, P. et al. Proc. Natl. Acad. Sci. USA 2005, 102, 4688; Williams, A.C. & Barry, B.W. Chemical permeation enhancement, In: Enhancement in Drug Delivery E., Touitou B.W., Barry (Eds.), CRC Press, Boca Raton, 2007, pp.233-254; Jampilek, J. et al. Med. Res. Rev., in press, DOI 10.1002/med.20227).

Transdermal therapeutic systems (TTSs) or transdermal drug delivery systems are topical dosage forms intended to deliver a drug substance at a controlled rate through the intact skin into the systemic circulation and to maintain efficacious plasma levels during prolonged time. A characteristic feature of TTSs in comparison with other topical dosage forms (ointments, creams) is the transport of defined and precise drug dosages through the healthy skin per defined time. The area-dosage-time relationships are determined as a crucial factor. Predecessors of TTSs were semi-solid topical dosage forms, mostly ointments, with the expected systemic effect. For some indications TTSs have already been used in clinical practice for the treatment of a variety of systemic diseases, for certain drugs they are tested or clinical trials are performed nowadays. At present transdermally applicable drugs include glycerol trinitrate, scopolamine, , oxybutynin, contraceptives, anodynes (e.g., , ), antihypertensive or antiarythmic drugs (e.g., , ), drugs against /inflammatory or antiparkinsonics (rotigotine). A number of new pharmaceuticals are developed or even under clinical evaluation, for example antipsychotics, non-steroidal hormones or antineoplastics (e.g., physostigmine, selegiline, insulin or 5-fluorouracil). However, the number of drugs that can be delivered transdermally is more limited than it was originally expected (Jampilek, J. et al. Med. Res. Rev., in press, DOI 10.1002/med.20227; Bos, J.D. et al. Exp. Dermatol. 2000, 9, 165; Benson, H.A.E. Curr. Drug Deliv. 2005, 2, 23; Delgado-Charro, M.B. & Guy, R.H. Transdermal drug delivery, In: Drug Delivery and Targeting, Hillery A.M., Lloyd A.W., Swarbrick J. (Eds.), Taylor & Francis, London, pp.207-236; Swart, P.J. et al. Int. J. Pharm. 1992, 88, 165; Muller, W. et al. U.S. Patent 7,413,747, 2008; Moller, H.J. et al. Pharmacopsychiatry 1999, 32, 99; Lee, K.C. et al. Neuropsychiatr. Dis. Treat. 2007, 3, 527; Wong, T.W. Recent Pat. Drug Deliv. Formul. 2009, 3, 8; Chandrashekar, N.S. et al. Asian Pac. J. Cancer Prev. 2008, 9, 437). The advantages of transdermal administration include, above all, good pharmacokinetic properties of application systems, the ability to maintain long-lasted stationary plasma levels of active substances, including drugs with short biological half-lives, which reduces undesirable side effects occurring as a result of considerable fluctuations of drug plasma levels. In contrast, the plasma levels achieved with the use of conventional dosage forms exhibit peaks and may even reach a toxic level leading to complications. Presystemic elimination of the applied dosage (hepatic first-pass effect) and such effects as pH change in the GIT or interactions with simultaneously applied preparations or food are also prevented effectively. TTSs also provide a possibility to apply drugs with a narrow therapeutic window and to interrupt drug delivery to the system immediately in the case of undesirable effect occurrence (in contrast to other conventional dosage forms, which do not provide such a possibility). An important advantage is very simple and painless application. TTSs are a non invasive alternative to parenteral, subcutaneous and intramuscular injections {Jampilek, J. et al. Med. Res. Rev., in press, DOI 10.1002/med.20227; Delgado-Charro, M.B. & Guy, R.H. Transdermal drug delivery, In: Drug Delivery and Targeting, Hillery A.M., Lloyd A.W., Swarbrick J. (Eds.), Taylor & Francis, London, pp.207-236; Rabiskova M. et al. Technology of Pharmaceutics, 3rd ed., Galen Prague, 2006).

Skin structure The skin is a human organ with the largest area and consists of three basic functional layers: the upper layer (epidermis), corium (dermis), and subcutaneous tissue (hypodermis). These layers localize capillaries, nerve endings and skin appendages (hair, nails and sebaceous, sweat and apocrine glands). The skin performs a number of different functions; the most important is protection from excessive water loss and mechanical, chemical, microbial and physical impacts. The most important is the outermost layer of epidermis, the horny layer (stratum corneum, SC), which is in fact the skin barrier. The structure of the SC can be described as a "bricks and mortar" model: corneocytes rich in keratin represent hydrophilic "bricks", and the lipid matrix represents hydrophobic "mortar" (Forslind, B. & Lindberg, M. Skin, Hair, Nails: Structure and Function. Marcel & Dekker New York, 2004; McGrath, J.A. et al. Rook's Textbook of Dermatology, 7th ed., Blackwell Publishing, 2004; Jampilek, J. et al. Med. Res. Rev., in press, DOI 10.1002/med.20227).

Routes of drug penetration through skin The penetration through the least permeable layer, the SC, is a limiting process. The live epidermis acts as a barrier only for penetration of extremely lipophilic compounds. There are three main possible routes for penetration of drug molecules through the intact skin or the SC: 0 appendageal route (transport via sebaceous and sweat glands and the transfollicular route - through hair follicles), ii) transcellular route (through corneocytes), Hi) intercellular route (through the intercellular space). In fact, the last two routes of drug penetration to the organism are considered as the most probable; together they are sometimes denominated as transepidermal (Karande, P. et al. Proc. Natl. Acad. Sci. USA 2005, 102, 4688; Williams, A.C. & Barry, B.W. Chemical permeation enhancement, In: Enhancement in Drug Delivery E., Touitou B.W., Barry (Eds.), CRC Press, Boca Raton, 2007, pp.233-254; Jampilek, J. et al. Med. Res. Rev., in press, DOI 10.1002/med.20227).

Affection of the skin barrier The skin barrier for facilitation of drug penetration can be affected by physical skin penetration enhancement techniques, including iontophoresis, electroporation; acoustical methods and microneedles. The disadvantage of these methods is their comparatively complicated use requiring special apparatus for their application and consequent financial cost; therefore mostly chemical transdermal penetration enhancers are used. At present the compounds interacting mainly with lipid components of SC or corneocytes are considered as transdermal chemical penetration enhancers (CPEs). These compounds are able to specifically affect intercellular space between corneocytes or modify corneocytes by hydration or denaturation of keratin, see details below (Karande, P. et al. Proc. Natl. Acad. Sci. USA 2005, 102, 4688; Williams, A.C. & Barry, B.W. Chemical permeation enhancement, In: Enhancement in Drug Delivery E., Touitou B.W., Barry (Eds.), CRC Press, Boca Raton, 2007, pp.233-254; Jampilek, J. et al. Med. Res. Rev., in press, DOI 10.10Q2/med.20227).

Transdermal chemical penetration enhancers; their mechanisms of action As with other pharmaceutical excipients, properties of CPEs are subject to strict requirements. An ideal enhancer should: i) be nontoxic, non-irritating and cause no allergic reactions; ii) have reversible influence on the skin barrier; after removal the skin should immediately and fully recover its normal barrier functions; i) possess rapid onset of action with a predictable and repeatable effect; iv) be pharmacologically and chemically inert; v) act selectively in one direction, i.e. ensuring API entrance to the body but preventing a loss of endogenic materials from the body; vi) be physically and chemically compatible with medicinal substances and other excipients in the preparation; vii) be cosmetically acceptable with convenient organoleptic properties; viii) have uncomplicated and rather inexpensive synthesis; ix) meet recently imposed additional biodegradability requirements. From the chemical point of view the group of CPEs is very heterogeneous. All CPEs were divided into 10 categories that covered the broad spectrum of evaluated compounds: i) anionic surfactants, ii) cationic surfactants, i) zwitterionic surfactants, iv) non-ionic surfactants, v) fatty acids, vi) sodium salts of fatty acids, vii) fatty , viii) fatty , ix) Azone-like compounds, and x) others (e.g., thioglycolate, , , cyclodextrins). Several possible mechanisms of action of enhancers were hypothesized but the exact mechanisms have not been elucidated. It is almost certain that they exhibit multiple effects. Generally, based on information from the section about the SC, it can be stated that: i) CPEs can interact with the intercellular lipid matrix (especially ceramides), or ii) they can interact with protein structures (influencing the conformation of keratin in the corneocytes or proteins in desmosomes), or i) CPEs can promote partitioning (influencing the SC nature leads to raising the penetrant concentration gradient and thus increasing the flux, i.e. increasing the concentration of the drug in the skin). A hypothesis was proposed that small polar molecules may break the intermolecular H-bonds that hold the ceramide molecules together in the SC. The hypothesis about the local molecular effect of CPEs on auto-associated ceramide films (breaking the H-bonds between ceramides and simultaneous incorporation of CPEs among them and creation of new H-bonds linking weakly the polar head groups of CPEs and ceramides) was confirmed by several vibrational spectroscopic studies. The attenuated total reflection (ATR) technique of Fourier-transform infrared spectrometry (FT-IR) and non-resonance Raman spectroscopy became popular techniques in characterization of the penetration activity of potential CPEs (Karande, P. et al. Proc. Natl. Acad. Sci. USA 2005, 102, 4688; Williams, A.C. & Barry, B.W. Chemical permeation enhancement, In: Enhancement in Drug Delivery E., Touitou B.W., Barry (Eds.), CRC Press, Boca Raton, 2007, pp.233-254; Jampilek, J. et al. Med. Res. Rev., in press, DOI 10.1002/med.20227, Zhao, J.et al. Real-time Raman spectroscopy for noninvasive in vivo skin analysis and diagnosis, In: New Developments in Biomedical Engineering. Campolo D. (Ed.), InTech, 2010). Alaptide

Alaptide, (S)-8-methyl-6,9-diazaspiro[4.5]decan-7,10-dione (see Fig. 1), is a compound discovered in the 1980s at Prague by Kasafirek et al. The substance preparation, production procedures and therapeutic application were protected by a number of patents in Czechoslovakia/the Czech Republic and abroad. Alaptide showed significant curative effect in different therapeutic areas on experimental animal models and also demonstrated very low acute toxicity in rats and mice. Furthermore, teratogenic and embryotoxic effects of alaptide were not observed. Evaluation of subchronic and chronic toxicity was carried out in rats in the dosage of 20 mg/ml and dogs in the dosage of 10 mg/ml, and no toxic effects were registered. Metabolic studies in rats showed that alaptide is excreted unchanged, mostly via urine (90%) {Kasafirek E. et al. Belg. Pat. 897843, 1984; CS Pat. 231227, 1986; Toxicol. Lett. 1986, 31, 189; CS Pat. 277132, 1992; CS Pat. 276270, 1992; Drugs Fut. 1990, 15, 445; Collect. Czech. Chem. Cormmun. 1992, 57, 179; Collect. Czech. Chem. Commun. 1993, 58,. 2987; US Pat. 5,318,973, 1994; Collect. Czech. Chem. Commun. 1994, 59, 195).

Figure 1. Structure of alaptide.

The knowledge of the structure and properties of CPEs, the hypotheses of CPE mechanism of action and our previous experience with several other groups of CPEs (Brychtova, K. et al. Bioorg. Med. Chem. 2010, 18, 73; Bioorg. Med. Chem. 2010, 18, 8556; Bioorg. Med. Chem. 2012, 20, 86; Mrozek, L. et al. 2011, 76, 1082; Jampilek, J. et al. Med. Res. Rev., in press, DOl 10.1002/med.20227) led us to the thought to evaluate alaptide as a potential transdermal penetration enhancer. In a number of in vitro tests it was demonstrated that alaptide is very effective in this respect. Based on the excellent enhancement activity, both alaptide were tested in metabolic and induction studies on primary human hepatocyte cultures. It was found that the alaptide enantiomers do not induce biotransformation CYPIAI, CYP1A2 and CYP1B1 in hepatocytes. These biotransformation enzymes are critical in bioactivation of procarcinogens (such as polychlorinated aromatic hydrocarbons or planar hydrocarbons) and are upregulated also by ultraviolet-B radiation (UVB) in the skin. CYPIAI is involved in the metabolic activation of aromatic hydrocarbons from pollution or industry contamination (polycyclic aromatic hydrocarbons such as benzopyrene, BP) by transforming it to BP-7,8-dihydrodiol-9,10- epoxide, which is the ultimate carcinogen. UVB-mediated induction of cytochromes CYP1A1 and CYP1B1 in human skin will probably result in enhanced bioactivation of polycyclic aromatic hydrocarbons and other environmental pollutants to which humans are exposed, which in turn could make the human skin more susceptible to UVB -induced skin cancers or allergic and irritant contact dermatitis {Jampilek, J. et al. Czech Patent Application PV 2011- 232, 2011; Jampilek, J. et al. Czech Patent Application PV 2011-495, 2011; Jampilek, J. et al. Czech Patent Application PV 2012-72, 2012; Jampilek, J. et al. Czech Patent Application PV 2012-511, 2012).

SUMMARY OF INVENTION:

The subject matter of the invention is the usage of alaptide with structure formula / as a transdermal penetration modifier of non-steroidal anti-inflammatory drugs (otherwise non steroidal antiphlogistics or NSA s) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/antimycobacterials, antimycotics, antivirotics) in pharmaceutical compositions (formulations) convenient for transdermal application.

/

The used non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) are sampled from the following list: , , , tribuzone, , , , clofezon, , , , , , , aclantate, , flunixine, ibufenac, , , , , tropesin, , , , , , fenbumetone, indobufene, , , , , , , , aceklofenac, , , , fentiazac, , , , , , , , flobufen, , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , prochazon, orgotein, , , , , , , , polysulfate, , pentosan polysulfate and , thus NSAIDs from the classes of salicylates, pyrrazolidindiones, fenamates, derivatives of arylalkanoic acids (fenacs, profens), oxicames, coxibes and other non-steroidal compounds, which mechanism of action is connected with inhibition of the metabolic pathway of and generation of pro-inflammatory substances such as , , and leukotrienes.

The used antipyretics/non-opiate analgesics are sampled from the following list: , , , , acetylsalicylic acid, choline salicylate, lysine salicylate, salicylate sodium or potassium, salicylate, morpholine salicylate, , , , , , methylsalicylate, , lysine acetylsalicylate, , gentisie acid, , , , , , ramiphenazone, , rimazoline, , , , , , , menthol, , other antipyretics/ non-opiate analgesics from the classes of anilines, salicylic acid derivatives and and other compounds affecting body thermoregulation or belonging to non-opiate analgesics.

The used glucocorticoids are sampled from the following list: , , , , , , , , , , , , methylprednisolone aceponate, , , , , betamethasone dipropionate, , flumethasone, , , , , prednylidene, , , triamcinolon acetonide, , , , , acetonide, , , furoate, propionate, other glucocorticoids from the classes of hydrocortisone, prednisolone, dexamethasone and triamcinolone and other compounds (steroidal anti-inflammatory drugs), which mechanism of action is connected with interference with the metabolism of arachidonic acid and with following inhibition of generation of pro-inflammatory icosanoids (prostacyclins, prostaglandins, thromboxanes, leukotrienes) and/or a decrease in production of and/or a decrease in histamine liberation.

The used antimicrobial chemotherapeutics are sampled from the following list: antibacterials: (antibiotics, antibacterial/antimycobacterial chemotherapeutics): classes of beta-lactam antibiotics (penicillins, carbapenems, monobactams and/or cephalosporins, carbacephems, oxacephems), macrolides, tetracyclines, aminoglycosides, polypeptides, glycopeptides, lincosamides, lipopeptides, ansamycins, fusidic acid, linezolid, classes of sulfonamides, quinolones, amphenicols, nitrofurans, nitroimidazoles, p-aminosalicylic acid, cycloserine, , pyrazinamide, ethionamide, protionamide, ethambutol, clofazimine, dapsone as well as other natural, semisynthetic or synthetic antibacterially effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of bacterial pathogens and causes their death. antimycotics : classes of polyenes, griseofulvin, , triazoles, allylamines and other non-azole ergosterol biosynthesis inhibitors, thiocarbamates, glucan synthesis inhibitors (echinocandines, pneumocandines, papulacandines), antimetabolites (flucytosine), ciclopirox, amorolfine and other antifungal effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of fungal pathogens and causes their death. antivirotics : classes of pyrimidine and purine , reverse transcriptase inhibitors, HIV-protease inhibitors, neuramidase inhibitors, amantadine, interferons, foscarnet as well as other antiviral effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of viral pathogens and causes their death.

Also the subject matter of the invention is the pharmaceutical composition for transdermal application containing anti-inflammatory drugs (non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids) and/or antimicrobial chemotherapeutics (antibacterials/ antimycobacterials, antimycotics, antivirotics) and simultaneously alaptide as a chemical transdermal penetration modifier.

DETAILED DESCRIPTION OF INVENTION:

In transdermal application alaptide causes an increase or a decrease, in dependence on the used supporting medium (pharmaceutical formulation), in absorption/penetration of non steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/antimycobacterials, antimycotics, antivirotics) to the skin and/or through the skin so that the concentration of the used drug increased at the place of administration, and/or the systemic concentration increased, or it was ensured that drugs act only on the skin surface/in the skin surface layer and do not penetrate to the skin deeper layers or do not have any systemic effects. The utilization of alaptide as a chemical modifier of transdermal penetration of non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/antimycobacterials, antimycotics, antivirotics), i.e. as an excipient, is completely unique, and only in this application this possibility of alaptide application is specified for the first time.

Also the subject matter of the invention are original pharmaceutical compositions for transdermal application containing non-steroidal anti-inflammatory drugs (otherwise non steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/antimycobacterials, antimycotics, antivirotics) and simultaneously containing alaptide as the chemical transdermal penetration modifier, which modifies the permeability of non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/antimycobacterials, antimycotics, antivirotics) through the skin and acts as a transdermal penetration enhancer or, by contrast, in dependence on the used pharmaceutical composition, acts as a penetration inhibitor and inhibits systemic effects of drugs.

Pharmaceuticals containing non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics are convenient both for local/topical and systemic treatment. Their mechanism of action is related to inhibition of the metabolic pathway of arachidonic acid and thus generation of pro-inflammatory substances such as prostacyclins, prostaglandins, thromboxanes and leukotrienes. In addition to anti inflammatory effect they decrease an elevated temperature, decrease pain perception, act antirheumatically and can have other effects (e.g., increase of uric acid or decrease of platelet agglutination). They can be used for treatment of mild or moderate pain (somatic pain, e.g., headache, toothache, , dysmenorrhoea, backache) and of different types (inflammations of the musculoskeletal system, e.g., rheumatic fever, rheumatoid , osteoarthrosis). The antiaggregant effect is used for prevention of acute cerebrovascular accident and myocardial infarction. Due to their effect they are used for treatment of various fevers.

Pharmaceuticals containing glucocorticoids are suitable for both local and systemic treatment. Corticoids are indicated for treatment of adults, adolescents as well as children. They express significant anti-inflammatory, anti-edematous and anti-pruritic effect and decrease hyperemia. Systemic treatment can be either substitution therapy - at adrenal insufficiency (prednisone, cortisole) or they can be indicated in specific cases, for example: allergic reactions and illnesses (e.g., bronchial asthma, including status asthmaticus, anaphylactic reactions, including after drug administration, posttransfusion reactions, contact dermatitis, serum sickness, larynx oedema, allergic rhinitis, etc.), autoimmune illnesses (e.g., rheumatoid polyarthritis), haematological illnesses (e.g., leukaemia), infections (e.g., sepsis by G- microbes), prevention of oedema of soft tissues (e.g., brain oedema), organ transplantation (e.g., prevention of rejection reaction), kindness illnesses (e.g., nephrotic syndrome) and skin illnesses (e.g., atopic dermatitis). Corticoids can be used for treatment of inflammatory manifestations and pruritus: eczemas (including atopic and discoid eczemas); dyshidrotic eczema; dermatitis Solaris, prurigo nodularis, psoriasis, neurodermatitis (including lichen simplex), lichen planus, seborrheic dermatitis, contact hypersensitivity manifestations and other dermatitises, lupus erythematodes chronicus discoides, generalized erythroderma, reactions on insect biting and heat rash. They can be indicated at brain oedema, both traumatogenic and connected with expansive process. Corticoids can also be indicated for therapy of aspiration pneumonia together with antibiotics, , collagenoses, nephrotic syndrome, lymphatic leukaemia, at infections (with appropriate ): disseminated or fulminant pulmonary tuberculosis and TB meningitis. Besides, they can be used as a part of treatment of shock states such as hemorrhagic, traumatic or septic shocks. Corticoids can be also used in the therapy of osteoarthrosis, carpal tunnel syndrome, synovitis, irritate arthritis, bursitis, uratic arthritis, epicondylitis, fibrositis and tendovaginitis.

Antimicrobial chemotherapeutics are compounds used at present in human and veterinary medicine for growth suppression or elimination of microorganisms (bacteria, yeasts and other fungi and viruses). Antimicrobial chemotherapeutics include antibiotics (compounds of natural origin or semisynthetically modified compounds), antibacterial chemotherapeutics (synthetic compounds), including antituberculotics, antimycotics and antivirotics. Antimicrobial chemotherapeutics are used primarily for treatment of various infectious states but sometimes also for prevention (antibiotic prophylaxis). Often a combination of some antibiotics and antimycotics is used to cover a broad spectrum of microorganisms (e.g., combination of penicillin, streptomycin and amphotericin B). These drugs are used for both local and systemic therapy. The subject matter of the invention is the usage of alaptide with structure formula / as a chemical modifier of transdermal penetration of non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/ antimycobacterials, antimycotics, antivirotics) in the pharmaceutical composition (formulation) suitable for transdermal application, which causes an increase or a decrease, in dependence on the used supporting medium (pharmaceutical formulation), in absorption/penetration of non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/antimycobacterials, antimycotics, antivirotics) to the skin and/or through the skin so that the concentration of the used drug increased at the place of administration, and/or the systemic concentration increased, or it was ensured that drugs act only on the skin surface / in the skin surface layer and do not penetrate to, the skin deeper layers or do not have any systemic effects. The utilization of alaptide as a chemical modifier of transdermal penetration of non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/ antimycobacterials, antimycotics, antivirotics), i.e. as an excipient, is completely unique, and only in this application this possibility of alaptide application is specified for the first time.

Alaptide was initially tested as an excipient that influences penetration of different compounds through the skin on the model drug theophylline, which average permeation in combination with micronized alaptide was increased by 65%. After that penetration of ibuprofen through the skin was tested from propylene glycol/water (1:1) medium. The addition of 1% of alaptide (in relation to ibuprofen amount) increased the penetration by 113% within 1 h and by 147% within 2 h. From hydroxypropyl cellulose gel without alaptide addition, max. 0.78% of ibuprofen penetrated through the skin within 2 h. The addition of 1% of alaptide (in relation to ibuprofen amount) increased the penetration by 177% within 1 h and by 246% within 2 h. The addition of 0.1% of alaptide (in relation to ibuprofen amount) increased the penetration of ibuprofen through the skin from hydroxypropyl cellulose gel by 177% after 30 min and from oleo-cream by 30% more than without alaptide. The penetration of nimesuHde from cream with the addition of 0.1% of alaptide was by 150% higher after 30 min and with the application of 0.1% of nanonized alaptide, by 80% more than from compositions without alaptide. The penetration of meloxicam with the addition of 0.1% of nanonized alaptide was by 383% higher after 30 min than without alaptide. The penetration of acetylsalicylic acid from propylene glycol/water medium with the addition of nanonized alaptide increased by 585% after 30 min, and the penetration of paracetamol with the addition of 0.1% of nanonized alaptide from propylene glycol/water medium was by 1657% higher in 120 min than without alaptide. The penetration of diclofenac with application of 0.1% of alaptide increased by 124% after 30 min and at application of nano-alaptide by 1445% in comparison with the penetration of diclofenac without alaptide. Diclofenac from carbomer gel with the application of 0.1% of nanonized alaptide penetrated by 77% more after 30 min than without alaptide, and from hydro-ointment with the addition of 0.1% of alaptide the penetration of diclofenac increased by 42% after 30 min. The penetration of diclofenac from hydroxypropyl cellulose gel with the addition of micronized alaptide increased by 55% after 60 min.

The penetration through the skin of the budesonide was tested from the propylene glycol/water (1:1) medium, from phosphate buffer (pH 7.4) and from isopropyl myristate without and with the presence of 0.1% of micronized alaptide (in relation to budesonide amount). The permeation of budesonide from the propylene glycol/water medium increased after the addition of alaptide by 480% within 8 h. After the addition of alaptide the penetration of budesonide from the buffer was on the average by 170% more within 20-24 h. After the addition of alaptide, budesonide permeation from isopropyl myristate increased on the average by 30% within 6-8 h and by 50% within 20-24 h. Dexamethasone penetration through the skin both from the buffer and the propylene glycol/water medium was possible within 30' min only after the addition of nanonized alaptide. The permeation of dexamethasone acetate with 0.1% amount of micronized alaptide (in relation to dexamethasone acetate amount) from the propylene glycol/water medium increased on the average by 110% within 6-8 h after the addition of alaptide. The amount of dexamethasone acetate penetrated from the buffer after the addition of alaptide was on the average by 190% more within 20-24 h. The permeation of dexamethasone acetate from isopropyl myristate after the addition of alaptide increased on the average by 20% within 20-24 h. Dexamethasone acetate penetration through the skin from carboxymethylcellulose gel with added 0.1% amount of micronized alaptide (in relation to dexamethasone acetate amount) was higher on the average by 10%. Without alaptide addition fluocinolone was not detectable within 3 h; by contrast, after the addition of nanonized alaptide fluocinolone permeation both from the buffer and the propylene glycol/water medium significantly increased after 30 min; in the case of the propylene glycol/water medium with nanonized alaptide the permeation even increased by 1100% within 1 h. After the addition of alaptide, the permeation of through the skin from the propylene glycol/water medium increased by 180% within 4-8 h and from the buffer by 250% within 8 h. Fluocinolone acetonide penetration from isopropyl myristate increased by 590% within 4 h after the addition of alaptide. At the concentration of 0.1% of alaptide in relation to the amount of fluocinolone acetonide in the formulation, the penetration of fluocinolone acetonide from hydro-cream was on the average by 260% more within 4-8 h, from oleo-cream by 230% more within 4—8 h and from the ointment on the average by 20% more within 20-24 h than without utilization of alaptide as a transdermal penetration modifier. After the addition of 0.1% of micronized alaptide (in relation to hydrocortisone acetate amount) hydrocortisone acetate penetrated through the skin from the propylene glycol/water medium within 4 h (without alaptide it did not penetrate at all). From the buffer hydrocortisone acetate penetrated on the average by 40% more within 20-24 h after the addition of alaptide. Hydrocortisone acetate permeation from isopropyl myristate after the addition of alaptide increased by 40% within 20-24 h than without alaptide. The penetration of prednisolone acetate through the skin from the propylene glycol/water medium increased by 300% within 20-24 h after the addition of alaptide. The amount of prednisolone acetate that permeated from the buffer after the addition of alaptide increased on the average by 270% within 4-8 h, and the permeation of prednisolone acetate from isopropyl myristate after the addition of alaptide increased on the average by 30% within 20-24 h. Prednisolone from the buffer with micronized alaptide did not permeate, but from the propylene glycol/water medium with micronized alaptide permeated after 90 min. After the addition of nanonized alaptide the permeation of prednisolone from both the buffer and the propylene glycol/water medium significantly increased after 30 min, in the case of the propylene glycol/water medium with nanonized alaptide even by 275% within 3 h. After the addition of 0.1% of micronized alaptide (in relation to prednisone amount), the permeation of prednisone from the propylene glycol/water medium increased by 680% within 4 h, the permeation of prednisone from the buffer increased by 650% within 4 h, and the permeation from isopropyl myristate increased by 320% within 4 h. Triamcinolone did not penetrate through the skin, but the addition of micronized alaptide to the buffer increased the penetration of triamcinolone by 174% after 30 min, and from the propylene glycol/water medium containing the micronized alaptide triamcinolone penetrated by 267% more after 30 min. Without the addition of alaptide penetrated through the skin from both media in 90 or 120 min. After the addition of micronized alaptide to the buffer, the penetration of triamcinolone acetonide increased by 5% in 90 min. The addition of nanonized alaptide to the propylene glycol/water medium enabled the penetration of triamcinolone acetonide already in 30 min, and the addition of micronized alaptide increased the penetration by 790% in 30 min compared to nanonized alaptide. Without the presence of alaptide triamcinolone acetonide from oleo-ointment was not detectable within 3 h, but after the addition of micronized alaptide triamcinolone acetonide was detected already after 30 min. The addition of nanonized alaptide increased the penetration of triamcinolone acetonide through the skin from the oleo-ointment by 273%. Without the presence of alaptide triamcinolone acetonide from hydro-cream was not detectable within 3 h. After the addition of micronized alaptide, triamcinolone acetonide from the cream was detected already after 90 min and with the addition of nanonized alaptide already after 1 h. The penetration of triamcinolone acetonide from the cream with nanonized alaptide increased by 124% after 90 min in comparison with micronized alaptide. Triamcinolone acetonide from methylcellulose gel without alaptide was not detectable within 3 h, but after the addition of micronized alaptide it was found already after 120 min. The addition of nanonized alaptide increased the penetration of triamcinolone acetonide from the gel by 228% i comparison with micronized alaptide. Triamcinolone acetonide from carbomer gel without alaptide was not .also detectable within 3 h. After the addition of micronized alaptide, triamcinolone acetonide from the gel was detected after 120 min. The addition of nanonized alaptide increased the penetration of triamcinolone acetonide from the gel by 5% compared to micronized alaptide.

The penetration of amoxicillin through the skin was evaluated from the propylene glycol/water (1:1) medium, from phosphate buffer (pH 7.4) and from isopropyl myristate without and with the presence of 0.1% amount of micronized alaptide (in relation to amoxicillin amount). The permeation of amoxicillin from the propylene glycol/water medium increased after the addition of alaptide by 55% within 8 h and on the average by 92% within 20-24 h. The penetration of amoxicillin from the buffer after the addition of alaptide was on the -average by 106% more within 8 h. The permeation of amoxicillin from isopropyl myristate after the addition of alaptide increased on the average by 5% within 8 h and by 30% within 20-24 h. The addition of alaptide increased the permeation of ampicillin from the propylene glycol/water medium by 145 % within 8 h. On the average the amount of ampicillin that penetrated from the buffer after the addition of alaptide was by 35% more within 8 h. The permeation of ampicillin from isopropyl myristate after the addition of alaptide increased on the average by 54% within 8 h. After the addition of alaptide, the permeation of oxacillin from the propylene glycol/water medium increased by 150% within 8 h and on the average by 80% within 20-24 h. The penetration of oxacillin from the buffer was on the average by 27% more within 8 h after the addition of alaptide. The permeation of oxacillin from isopropyl myristate after the addition of alaptide increased on the average by 177% within 8 h. The permeation of benzylpenicillin (penicillin G) from the propylene glycol/water medium increased by 136% within 8 h after the addition of alaptide. The permeation of penicillin G from the buffer after the addition of alaptide was on the average by 16% more within 24 h. Its permeation from isopropyl myristate increased on the average by 5% within 20-24 h after the addition of alaptide. The addition of alaptide increased the permeation of phenoxymethylpenicillin (penicillin V) from the propylene glycol/water medium by 56% within 8 h and on the average by 45% within 20-24 h. The permeation of penicillin V from the buffer after the addition of alaptide was on the average by 43% more within 8 h. The permeation of penicillin V from isopropyl myristate after the addition of alaptide increased on the average by 34% within 8 h. The permeation of ofloxacin from the propylene glycol/water medium after the addition of alaptide increased by 54% within 8 h. The addition of alaptide increased the penetration of ofloxacin from the buffer on the average by 137% within 8 h more and by 85% within 20-24 h. The permeation of ofloxacin from isopropyl myristate after the addition of alaptide increased on the average by 36% within 8 h. The penetration of ofloxacin through the skin from methylcellulose and carbomer gels was also tested without and with the presence of 0.1% of micronized alaptide in time. The penetration of ofloxacin from methylcellulose gel increased after the addition of alaptide on the average by 1040% within 4-8 h and by 136% within 20-24 h. The addition of alaptide increased the penetration of ofloxacin from carbomer gel on the average by 200% within 4-8 h and by 80% within 20-24 h. Also the penetration of sulfathiazole through the skin from oleo-ointment was tested without and with 0.1% (in relation to sulfathiazole amount) of micronized alaptide or nanonized alaptide in time. The penetration of sulfathiazole from oleo-ointment increased on the average by 10% within 8 h and by 37% within 12-24 h after the addition of micronized alaptide. The penetration of sulfathiazole from oleo-ointment increased on the average by 180% within 8 h and by 330% within 12-24 h after the addition of nanonized alaptide. The penetration of chloramphenicol through the skin from the propylene glycol/water medium and from the buffer was tested without and with 0.1% of micronized alaptide in time. The addition of micronized alaptide to the propylene glycol/water medium increased the penetration of chloramphenicol by 7% within 24 h and the addition to the buffer, by 1% within 24 h than without alaptide. The penetration of chloramphenicol through the skin from oleo-ointment was also tested without and with 0.1% (in relation to chloramphenicol amount) of micronized alaptide or nanonized alaptide in time. The penetration of chloramphenicol from the oleo-ointment increased after the addition of micronized alaptide on the average by 90% within 8 h and after the addition of nanonized alaptide on the average by 360% within 8 h and by 1030% within 24 h. The permeation of neomycin sulfate through the skin from the propylene glycol/water medium without and with 0.1% of micronized alaptide was evaluated in time. After the addition of alaptide the permeation of neomycin sulfate increased by 165% after 30 min. The permeation of mupirocin through the skin from hydro-ointment was increased by added 0.1% amount of micronized alaptide by 144% already after 30 min and approx. by 400% after 60 min; thus alaptide significantly accelerated the penetration of mupirocin through the skin. On the contrary, the addition of nanonized alaptide significantly inhibited penetration through the skin (decreased the penetration by 60% during the 1st hour); thus mupirocin will act only on the surface of the skin. The addition of micronized alaptide to the buffer increased the permeation of pyrazinamide from by 63% after 8 h and by 303% after 24 h, and the addition of nanonized alaptide increased the permeation of pyrazinamide by 125% within 24 h. Also the penetration of pyrazinamide through the skin from carbomer gel was tested without and with the presence of 0.1% (in relation to pyrazinamide amount) of micronized alaptide or nanonized alaptide in time. The addition of micronized alaptide to the gel increased the penetration of pyrazinamide by 16% within 24 h, and the addition of nanonized alaptide to the gel increased the penetration of pyrazinamide on the average by 16% within 24 h. The permeation of fluconazole through the skin from the propylene glycol/water medium after the addition of 0.1% of nanonized alaptide increased by 59% within 24 h. The penetration of fluconazole from oleo-ointment after the addition of nanonized alaptide increased on the average by 150% within 8 h and by 350% within 12-24 h. The addition of nanonized alaptide increased the penetration of fluconazole from cream by 28% within 12 h in comparison with the formulation without alaptide. The addition of micronized alaptide to the propylene glycol/water medium containing aciclovir caused an increase in the penetration of aciclovir by 114% already after 30 min, and the addition of nanonized alaptide to the propylene glycol/water medium containing aciclovir increased the penetration of aciclovir on the average by 158% after 30 min and by 280% after 2 h. The addition of micronized alaptide to the buffer increased the permeation of aciclovir by 126% already after 30 min and by 440% after 2 h. The addition of 0.1% of nanonized alaptide increased the penetration of aciclovir through the skin from hydro-cream by 25% already after 30 min, and the addition of 0.1% of micronized alaptide increased the penetration of aciclovir from carbomer gel on the average by 37% after 30 min.

The particle size distribution of the used micronized alaptide was 50-80% up to 10 maxium Feret diameters. It was measured by a microscope NIKON Optiphot 2 with a digital camera VDS CCD-1300F.

The used nanonized alaptide was prepared using a nanomill NETZSCH with glass beads. The particle size of nanonized alaptide was measured using Sympatec NANOPHOX equipment, and the particle size 5o- 90 was up to 900 nm.

Alaptide as the excipient affecting the penetration of drugs to/through the skin can be combined in pharmaceutical formulations with the following non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs): phenylbutazone, oxyphenbutazone, kebuzone, tribuzone, sulfinpyrazone, azapropazone, mofebutazone, clofezon, suxibuzone, flufenamic acid, niflumic acid, mefenamic acid, tolfenamic acid, meclofenamic acid, aclantate, etofenamate, flunixine, ibufenac, alclofenac, diclofenac, indometacin, acemetacin, tropesin, sulindac, lonazolac, tolmetin, ketorolac, nabumetone, fenbumetone, indobufene, zomepirac, bumadizone, etodolac, fentiazac, difenpiramide, oxametacin, proglumetacin, aceklofenac, bufexamac, felbinac, bendazac, fentiazac, nifenazone, ibuprofen, naproxen, ketoprofen, suprofen, flurbiprofen, pirprofen, flobufen, fenoprofen, fenbufen, benoxaprofen, indoprofen, oxaprozin, ibuproxam, dexibuprofen, flunoxaprofen, alminoprofen, dexketoprofen, vedaprofen, carprofen, tepoxalin, tiaprofenic acid, tenoxicam, droxicam, lornoxicam, piroxicam, meloxicam, isoxicam, nimesulide, celecoxib, rofecoxib, valdecoxib, parecoxib, etoricoxib, lumiracoxib, firocoxib, robenacoxib, mavacoxib, cimicoxib, prochazon, orgotein, feprazone, diacerein, morniflumate, tenidap, oxaceprol, glucosamine, benzydamine, glycosaminoglycan polysulfate, chondroitin sulfate, pentosan polysulfate, aminopropionitrile; thus NSAIDs from the classes of salicylates, pyrrazolidindiones, fenamates, derivatives of arylalkanoic acids (fenacs, profens), oxicames, coxibes and other non-steroidal compounds, which mechanism of action is connected with inhibition of the metabolic pathway of arachidonic acid and generation of pro-inflammatory substances such as prostacyclins, prostaglandins, thromboxanes and leukotrienes.

Alaptide can be also combined in pharmaceutical formulations with the following antipyretics/non-opiate analgesics: paracetamol, bucetin, propacetamol, salicylic acid, acetylsalicylic acid, choline salicylate, lysine salicylate, salicylate sodium or potassium, , morpholine salicylate, salsalate, ethenzamide, guacetisal, carbasalate calcium, salicylamide, methylsalicylate, aloxiprin, lysine acetylsalicylate, benorilate, gentisic acid, dipyrocetyl, diflunisal, phenazone, aminophenazone, propyphenazone, ramiphenazone, metamizole, rimazoline, glafenine, floctafenine, viminol, nefopam, flupirtine, ziconotide, menthol, nabiximols, other antipyretics/non-opiate analgesics from the classes of anilines, salicylic acid derivatives, pyrazolones and other compounds affecting body thermoregulation or belonging to non-opiate analgesics.

Alaptide can be also combined in pharmaceutical formulations with the following glucocorticoids: hydrocortisone, hydrocortisone acetate, fludrocortisone, tixocortol, medrysone, dexamethasone, dexamethasone acetate, prednisone, prednisolone, prednisolone acetate, fluprednisolone, methylprednisolone, methylprednisolone aceponate, fluorometholone, difluprednate, mazipredone, betamethasone, betamethasone dipropionate, paramethasone, flumethasone, desoximetasone, fluocortolone, diflucortolone, clocortolone, prednylidene,- fluprednidene, triamcinolone, triamcinolon acetonide, flunisolide, desonide, prednicarbate, budesonide, fluocinolone acetonide, fluocinonide, ciclesonide, mometasone furcate, , other glucocorticoids from the classes of hydrocortisone, prednisolone, dexamethasone, triamcinolone and other steroid compounds (steroidal anti inflammatory drugs), which mechanism of action is connected with interference to the metabolism of arachidonic acid and with following inhibition of generation of pro¬ inflammatory icosanoids (prostacyclins, prostaglandins, thromboxanes, leukotrienes) and/or a decrease in production of cytokines and/or a decrease in histamine liberation.

Alaptide can be also combined in pharmaceutical formulations with the following antimicrobial chemotherapeutics: ' ' ' antibacterials : - (antibiotics, antibacterial/antimycobaeterial chemotherapeutics): classes of beta-lactam antibiotics- (penicillins, carbapenems, monobactams- and/or cephalosporins, carbacephems, oxacephems), macrolides, tetracyclines, aminoglycosides, polypeptides, glycopeptides, lincosamides, lipopeptides, ansamycins, fusidic acid, linezolid, classes of sulfonamides, qiiinolones, amphenicols, nitrofurans, nitroimidazoles, ^-aminosalicylic acid, cycloseriiie, isoniazid, pyrazinamide, ethionamide, protionamide, ethambutol, clofazimine, dapsone as well as other natural, semisynthetic or synthetic antibacterially effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of bacterial pathogens and causes their death. aritimvcotics: class of polyenes, griseofulvin, imidazoles, triazoles, allylamines and other non- azole ergosterol biosynthesis inhibitors, thiocarbamates, glucan synthesis inhibitors (eehinocandines, pneumocandines, papulacandines), antimetabolites (flucytosine), ciclopirox, amorolfine and other antifungal effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of fungal pathogens and causes their death. antivirotics: class of pyrimidine and purine nucleotides, reverse transcriptase inhibitors, HIV- protease inhibitors, neuramidase inhibitors, amantadine, interferons and foscarnet as well as other antiviral effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of viral pathogens and causes their death.

Also the subject matter of the invention is the original pharmaceutical compositions for human and/or veterinary applications that are characterized by the combination of alaptide as an excipient with non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics in ointment, cream, gel or a transdermal therapeutic system, where alaptide modifies the permeability of non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) arid/or antipyretics/non-opiate analgesics through the skin and effects as a transdermal penetration modifier. The following non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) can be used: phenylbutazone, oxyphenbutazone, kebuzone, tribuzone, sulfinpyrazone, azapropazone, mofebutazone, clofezon, suxibuzone, flufenamic acid, niflumic acid, mefenamic acid, tolfenamic acid, meclofenamic acid, aclantate, etofenamate, flunixine, ibufenac, alclofenac, diclofenac, indometacin, acemetacin, tropesin, sulindac, lonazolac, tolmetin, ketorolac, nabumetone, fenbumetone, indobufene, zomepirac, bumadizone, etodolac, fentiazac, difenpiramide, oxametacin, proglumetacin, aceklofenac, bufexamac, felbinac, bendazac, fentiazac, nifenazone, ibuprofen, naproxen, ketoprofen, suprofen, flurbiprofen, pirprofen, flobufen, fenoprofen, fenbufen, benoxaprofen, indoprofen, oxaprozin, ibuproxam, dexibuprofen, flunoxaprofen, alminoprofen, dexketoprofen, vedaprofen, carprofen, tepoxalin, tiaprofenic acid, tenoxicam, droxicam, lornoxicam, piroxicam, meloxicam, isoxicam, nimesulide, celecoxib, rofecoxib, valdecoxib, parecoxib, etoricoxib, lumiracoxib, firocoxib, robenacoxib, mavacoxib, cimicoxib, prochazon, orgotein, feprazone, diacerein, morniflumate, tenidap, oxaceprol, glucosamine, benzydamine, glycosaminoglycan polysulfate, chondroitin sulfate, pentosan polysulfate, aminopropionitrile; thus NSAIDs from the classes of salicylates, pyrrazolidindiones, fenamates, derivatives of arylalkanoic acids (fenacs, profens), oxicames, coxibes and other non-steroidal compounds, which mechanism of action is connected with inhibition of the metabolic pathway of arachidonic acid and generation of pro-inflammatory substances such as prostacyclins, prostaglandins, thromboxanes and leukotnenes. The following antipyretics/non-opiate analgesics can be used: paracetamol, bucetin, propacetamol, salicylic acid, acetylsalicylic acid, choline salicylate, lysine salicylate, salicylate sodium or potassium, imidazole salicylate, morpholine salicylate, salsalate, ethenzamide, guacetisal, carbasalate calcium, salicylamide, methylsalicylate, aloxiprin, lysine acetylsalicylate, benorilate, gentisic acid, dipyrocetyl, diflunisal, phenazone, aminophenazone, propyphenazone, ramiphenazone, metamizole, rimazoline, glafenine, floctafenine, viminol, nefopam, flupirtine, ziconotide, menthol, nabiximols, other antipyretics/non-opiate analgesics from the classes of anilines, salicylic acid derivatives, pyrazolones and other compounds affecting body thermoregulation or belonging to non-opiate analgesics.

Also the subject matter of the invention is original pharmaceutical compositions for human and/or veterinary applications that are characterized by the combination of alaptide as an excipient and glucocorticoids in ointment, cream, gel or a transdermal therapeutic system, where alaptide modifies the permeability of glucocorticoids through the skin and acts as a transdermal penetration modifier. The following glucocorticoids can be used: hydrocortisone, hydrocortisone acetate, fludrocortisone, tixocortol, medrysone, dexamethasone, dexamethasone acetate, prednisone, prednisolone, prednisolone acetate, fluprednisolone, methylprednisolone, methylprednisolone aceponate, fluorometholone, difluprednate, mazipredone, betamethasone, betamethasone dipropionate, paramethasone, flumethasone, desoxirrietasone, fluocortolone, diflucortolone, clocortolone, prednylidene, fluprednidene, triamcinolone, triamcinolon acetonide, flunisolide, desonide, prednicarbate, budesonide, fluocinolone acetonide, fluocinonide, ciclesonide, mometasone furoate, clobetasol propionate, other, glucocorticoids from the classes of hydrocortisone, prednisolone, dexamethasone, triamcinolone and other steroid compounds (steroidal anti-inflammatory drugs), which mechanism of action is connected with interference to the metabolism of arachidonic acid and with following inhibition of generation of pro-inflammatory icosanoids (prostacyclins, prostaglandins, thromboxanes, leukotrienes) and/or a decrease in production of cytokines and/or,a decrease in histamine liberation. , , , Also the subject matter of the invention is original pharmaceutical compositions for. human and/or veterinary applications that are characterized by the combination of alaptide as an excipient and antimicrobial chemotherapeutics (antibacterials/antimycobacterials, antimycotics, antivirotics) in ointment, cream, gel or a transdermal therapeutic system, where alaptide modifies the permeability of antimicrobial chemotherapeutics (antibacterials/ antimycobacterials, antimycotics, antivirotics) through the skin and acts as a transdermal penetration modifier. The following glucocorticoids can be used: antibacterials : (antibiotics, antibacterial/antimycobacterial chemotherapeutics): classes of beta-lactam antibiotics (penicillins, carbapenems, monobactams and/or cephalosporins, carbacephems, oxacephems), macrolides, tetracyclines, aminoglycosides, polypeptides, glycopeptides, lincosamides, lipopeptides, ansamycins, fusidic acid, linezolid, classes of sulfonamides, quinolones, amphenicols, nitrofurans, nitroimidazoles, ^-aminosalicylic acid, cycloserine, isoniazid, pyrazinamide, ethionamide, protionamide, ethambutol, clofazimine, dapsone as well as other natural, semisynthetic or synthetic antibacterially effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of bacterial pathogens and causes their death. antimycotics : class of polyenes, griseofulvin, imidazoles, triazoles, allylamines and other non- azole ergosterol biosynthesis inhibitors, thiocarbamates, glucan synthesis inhibitors (echinocandines, pneumocandines, papulacandines), antimetabolites (flucytosine), ciclopirox, amorolfine and other antifungal effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of fungal pathogens and causes their death. antivirotics : class of pyrimidine and purine nucleotides, reverse transcriptase inhibitors, HIV- protease inhibitors, neuramidase inhibitors, amantadine, interferons and foscarnet as well as other antiviral effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of viral pathogens and causes their death.

Alaptide alone is poorly soluble; its solubility in water is 0.1104 g/100 mL, in ethanol

0.1011 g/100 mL and in the mixture watenethanol 1:1 0.3601 g/100 mL; its log P /o is 1.39. Alaptide can be effectively dissolved in aqueous solutions containing surfactants such as Tween 20, Tween 80, Macrogol 4000, Macrogol 6000, propylene glycol, sodium lauryl sulfate, poloxamer (Pluronic), castor oil polyethylene glycol ether (Cremophor EL) or various PEG-derivatives (PEG-stearates, PEG-esters of fatty acids, PEG-derivatives of fatty acid glycerides, PEG-D-a -tocoferole) or complexing compounds such as cyclodextrins and their derivatives (e.g., hydroxypropyl - -cyclodextrin), dextrans and their derivatives, pectins and their salts and derivatives, glucans and their derivatives, chitosan and its derivatives, methylcellulose and its salts and derivatives. The generation of such complexes/adducts results in the increase of alaptide solubility in water. The utilization of such complexes/adducts seems to be very advantageous for preparation of pharmaceutical compositions (formulations) for application. Complexes/adducts can be prepared by mixing aqueous solutions of surfactants or complexing agents with alaptide. After completion of mixing a complex/adduct can be used for preparation of pharmaceutical compositions (formulations), or the solvent can be evaporated, and the obtained evaporation solid residue (a product of complexation) can be consequently used for the preparation of pharmaceutical compositions (formulations). Thus alaptide can be applied alone or together with other excipients or a combination of excipients that increase its solubility in ointment, cream, gel or a transdermal therapeutic system. Alaptide can be used in the concentration from 0.001 to 5% as a chemical transdermal penetration enhancer that supports an increase in absorption/penetration of non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/antimycobacterials, antimycotics, antivirotics) to the skin and/or through the skin, increasing their concentration at the place of administration and/or their systemic concentration.

Similarly to the utilization of alaptide complexes/adducts with surfactants or complexing agents (see above), the preparation of nanoparticles of alaptide can be used. Alaptide nanoparticles can be obtained by milling alaptide with emulsifiers and other stabilizers. Alaptide nanoparticles were generated by dispergation using a technique of wet milling in the aqueous solution of a surface modifier. The suspension generated in this manner is milled using a ball mill in the presence of a milling medium. This method assumes pulverization of all major micrometer particles to nanoparticles. For example, deoxycholate sodium, sodium lauryl sulfate, poloxamer, povidone, Macrogol 6000 can be used as wetting agents. Grinding balls can be polystyrene, ceramic or glass.

Modified alaptide was obtained by the above-mentioned routes and showed considerably higher solubility and modified physico-chemical properties depending on modification, thus optimized for the particular composition of ointment (oleo-ointment, hydro-ointment), cream (oleo-cream, hydro-cream), gel or a transdermal therapeutic system. Alaptide alone (micronized alaptide) or nanonized alaptide or alaptide complexes/adducts can be used as pharmaceutical adjutants (excipients) for pharmaceutical compositions (formulations) designed for humane and/or veterinary applications. In this case alaptide serves as a chemical transdermal penetration modifier that influences the absorption/penetration of non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics (antibacterials/antimycobacterials, antimycotics, antivirotics) to the skin and/or through the skin, increasing their concentration at the place of administration and/or their systemic concentration.

According to bases, semisolid formulations can be generally divided into oleo-/hydro- ointments, oleo-/hydro-creams and gels. Untreated, surface-modified and nanonized alaptide was added to ointment, cream and gel bases in the amount ranged from 0.1% to 5% of the total composition of formulation.

In hvdrofobic oleo-ointments the following excipients can be used: • mixture of white or yellow vaseline and hydrated wool wax (lanolin, cera lanae hydrpsa), • mixture of white or yellow vaseline, liquid paraffin and wax (cera) or hydrated wool wax (lanolin, cera lanae hydrosa), • mixture of sunflower oil and stabilized lard and hydrated wool wax (lanolin, cera lanae hydrosa), • mixture of cetyl , white wax (cera), stabilized lard and hydrated wool wax (lanolin, cera lanae hydrosa), • mixture of dimethicones (polydimethylsiloxanes), hydrated wool wax (lanolin, cera lanae hydrosa) or cetyl alcohol or glycerol monostearate and isopropyl myristate in vaseline (white or yellow), liquid paraffin or vegetable oils.

In hydro-ointments mixtures of low- and high-molecular macrogols, e.g., 300 and 1500 (1:1), can be used as excipients.

In oleo-creams the following excipients can be used: • mixture of white or yellow vaseline, liquid paraffin, solid paraffin, wool wax (cera lanae), beeswax, stabilized lard (e.g., commercially available base Synderman®), • mixture of white or yellow vaseline, liquid paraffin, solid paraffin, wool wax (cera lanae), aluminium stearate (e.g., commercially available base Pontin®), • mixture of beeswax, liquid paraffin, solid paraffin, wool wax (cera lanae), zinc stearate, aluminium stearate, purified water, methylparaben, propylparaben, sodium tetraborate and Arlacel™ 481 (sorbitan oleate, hydrogeneted castor oil, beeswax, stearic acid), (e.g., commercially available base Cutilan®).

In hydro-creams the following excipients can be used: • mixture of white or yellow vaseline, liquid paraffin, cetylstearylalcohol (anionic emulsifying ointment according to the Czech Pharmacopoeia) and purified water, methylparaben and propylparaben (anionic cream according to the Czech Pharmacopoeia), • mixture of white or yellow vaseline, liquid paraffin, cetylstearylalcohol and Polysorbate 60 (non-ionic emulsifying ointment according to the Czech Pharmacopoeia) and propylene glycol and purified water, methylparaben and propylparaben (non-ionic cream according to the Czech Pharmacopoeia), • mixture of monoglycerides or of fatty acids and ethoxylolated fatty or ethoxylated fatty acid esters or ethoxylated fatty acid esters and sorbitan and mixture of antimicrobial compounds (e.g., commercially available base Neo-Aquasorb ®).

In gels the following excipients can be used: • mixture of methylcellulose, glycerol 85%, purified water (methylcellulose mucilage according to the Czech Pharmacopoeia) and methylparaben and propylparaben, • mixture of carboxymethylcellulose sodium, glycerol or sorbitol or propylene glycol, purified water, methylparaben and propylparaben, • mixture of liquid paraffin, solid paraffin, stearyl alcohol, propylene glycol, Slovasol™ 2430, polyacrylates (Carbomers), trolamine, purified water, methylparaben and propylparaben (e.g., commercially available base Ambiderman®).

Example of the composition of oleo-ointment with alaptide 0.01 to 100 % w/w (in relation to drug) and 1 to 10% of drug can be as follows: alaptide from 0.001 to 10 g, drug from 1 to

10 g, cera lanae hydrosa from 65 to 75 g, yellow vaseline from 10 to 20 g, liquid paraffin up to 100 g. (i.e. ointment base for antibiotics).

Example of the composition of hydro-ointment with alaptide 0.01 to 100 % w/w (in relation to drug) and 1 to 10% of drug can be as follows: alaptide from 0.001 to 10 g, drug from 1 to 10 g, macrogol up to 100 g. Example of the composition of oleo-cream with alaptide 0.01 to 100 % w/w (in relation to drug) and 1 to 10% of drug can be as follows: alaptide from 0.001 to 10 g, drug from 1 to 10 g, Synderman®from 80 to 95 g, propylene glycol up to 100 g.

Example of the composition of hydro-cream with alaptide 0.01 to 100 % w/w (in relation to drug) and 1 to 10% of drug can be as follows: alaptide from 0.001 to 10 g, drug from 1 to 10 g, Cremor Neo-Aquasorbi®from 80 to 95 g, propylene glycol up to 100 g.

Example of the composition of gel with alaptide 0.01 to 100 % w/w (in relation to drug) and 1 to 10% of drug can be as follows: alaptide from 0.001 to 10 g, drug from 1 to 10 g, Carboxymethylcellulose ointment (carboxymethylcellulose sodium 5 g, Macrogol 300 10 g, propylene glycol 2.5 g, methylparaben 0.2 g, propylparaben 0.2 g, purified water 87.3 g) up to 100 g.

This approach is described in detail in the following examples.

The particle size distribution of the used micronized alaptide was 50-80% up to 10 maxium Feret diameters. It was measured by a microscope NIKON Optiphot 2 with a digital camera VDS CCD-1300F.

The used nanonized alaptide was prepared using a nanomill NETZSCH with glass beads. The particle size of the used nanonized alaptide was measured by NANOPHOX (0138 P)

Sympatec equipment, and the particle size x5 0 -X9 0 was up to 900 nm.

In vitro experiments on penetration of drugs or formulations without or with the presence of alaptide as a chemical transdermal penetration modifier was performed using a vertical Franz diffusion cell ( volume of 5.2 mL and donor surface area of 0.635 cm2, SES-Analysesysteme, Germany). Full thickness dorsal skin from porcine ear (Sus scrofa f . domestica) was used as a model membrane. The primary screening in the presence of alaptide as a chemical transdermal penetration modifier was performed with theophylline. This drug is commonly used as a model compound at penetration experiments, because it has medium polarity and by itself penetrates through the skin very poorly.

BRIEF DESCRIPTION OF DRAWINGS:

Fig. 1; Influencing biotransformation enzymes CYP1A1, CYP1A2 and CYP1B1 in hepatocytes. Fig. 2: Penetration of theophylline (TEO) through the skin from water depending on the amount of micronized alaptide (ALA) in time. Of the original amount of TEO, max. 0.24% penetrated without the addition of ALA within 1 h. The addition of 1 mg ALA did not have any significant effect on TEO permeation, but the addition of 10 mg of ALA increased permeation through the skin approx. 1.5 times within 1 h and approx. 1.7 times within 2 h. After 24 h the effect of higher ALA amount was not apparent.

Fig. 3: Penetration of theophylline (TEO) through the skin from the phosphate buffer (pH 7.4) depending on the amount of micronized alaptide (ALA) in time. The addition of 1 mg of ALA did not cause an increase in TEO penetration within the short time interval of 2 h, but already from the 4th hour TEO penetration was higher by 15% and after 24 h it was higher by 63%. The addition of 10 mg of ALA to the system increased the average penetration of TEO by 65%.

Fig. 4: Penetration of theophylline (TEO) through the skin from the propylene glycol (PG)/water (1:1) medium depending on the amount of micronized alaptide (ALA) in time. The addition of 1 mg of ALA to the system increased the penetration of TEO by 35% within 2 h, but then penetration decreased and after 24 h it was higher only by 10%. The addition of 10 mg of ALA to the system increased the average penetration by approx. 180% within 1 h, and then the penetration decreased again.

Fig. 5: Penetration of ibuprofen (IBU) through the skin from the propylene glycol (PG)/water (1:1) medium depending on the amount of micronized alaptide (ALA) in concentrations 0.1%, 1%, 10%. Of the original amount of the used IBU, max. 0.36% penetrated without ALA within 2 h. The addition of 1 mg of ALA increased IBU penetration by 20%. The highest penetration was detected after the addition of 10 mg (1%) of ALA, the permeation was increased by 113% within 1 h and by 147% within 2 h. The further addition of ALA up to 100 mg (10%) of ALA decreased the penetration of IBU.

Fig. 6: Penetration of ibuprofen (IBU) through the skin from hydroxypropyl cellulose gel depending on the amount of micronized alaptide (ALA) in time. The hydroxypropyl cellulose gel with 1% of IBU was prepared, and micronized alaptide (ALA) in concentrations 0.1%, 1%, 10% was added. Of the original amount of IBU, max. 0.78% penetrated without ALA within 2 h. The highest penetration was detected after the addition of 10 mg (1%) of ALA; the permeation was increased by 177% within 1 h and by 246% within 2 h. The further addition of ALA up to 100 mg (10%) increased IBU penetration by 74% within 2 h.

Fig. 7 : Comparison of the penetration of ibuprofen (IBU) through the skin from various media without and with the presence of 0.1% (in relation to IBU amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from hydroxypropyl cellulose gel and from oleo-cream. The addition of ALA to gel increased the penetration of IBU through the skin by 177% within 30 min, and the addition of ALA to oleo-cream increased the penetration of IBU by 30%.

Fig. 8: Comparison of the penetration of nimesulide (NEVI) through the skin from various media without and with the presence of 0.1% (in relation to NIM amount) of micronized or nanonized alaptide (ALA, NALA) in time: from buffer (pH 7.4) and

from oleo-cream. The penetration of NEVI from cream with ALA increased by 150% ' ' within 30 min, and the penetration with NALA was by 80% more than from the formulation without NALA.

Fig. 9: Comparison of the penetration of meloxicam (MEL) through the skin from buffer (pH 7.4) without and with the presence of 0.1% (in relation to MEL amount) of micronized or nanonized alaptide (ALA, NALA) in time. The addition of NALA increased the permeation of MEL by 383% within 3 min.

Fig. 10: Comparison of the penetration of acetylsalicylic acid (ASA) through the skin from buffer (pH 7.4) or from the propylene glycol (PG)/water (1:1) medium without and with the presence of 0.1% (in relation to ASA amount) of micronized or nanonized alaptide (ALA, NALA) in time. The addition of NALA increased the permeation of ASA from PG/water medium by 585% within 30 min.

Fig. 11: Comparison of the penetration of paracetamol (PAR) through the skin from buffer (pH 7.4) or from the propylene glycol (PG)/water (1:1) medium without and with the presence of 0.1% (in relation to PAR amount) of micronized or nanonized alaptide (ALA, NALA) in time. The addition of NALA increased the permeation of PAR from PG/water medium by 1657% within 120 min.

Fig. 12: Comparison of the penetration of diclofenac (DIC) through the skin from buffer (pH 7.4) or from the propylene glycol (PG)/water (1:1) medium without and with the presence of 0.1% (in relation to DIC amount) of micronized or nanonized alaptide (ALA, NALA) in time. The addition of ALA increased the penetration of DIC by 124%, the addition of NALA by 1445%.

Fig. 13: Comparison of the penetration of diclofenac (DIC) through the skin from carbomer gel without and with the presence of 0.1% (in relation to DIC amount) of micronized or nanonized alaptide (ALA, NALA) in time. The addition of NALA increased the permeation of DIC from gel by 77% within 30 min.

Fig. 14: Comparison of the penetration of diclofenac (DIC) through the skin from hydroxypropyl cellulose gel without and with the presence of 0.1% (in relation to DIC amount) of micronized alaptide (ALA) in time. The addition of ALA increased the permeation of DIC from gel by 55% within 60 min.

Fig. 15: Comparison of the penetration of diclofenac (DIC) through the skin from hydro- ointment without and with the presence of 0.1% (in relation to DIC amount) of micronized alaptide (ALA) in time. The addition of ALA increased the permeation of DIC from hydro-ointment by 42% within 30 min.

Fig. 16: Comparison of the penetration of budesonide (BUD) through the skin from various media without and with the presence of 0.1% (in relation to BUD amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (IPM). Without the addition of ALA BUD (lO mg/mL, 100%) was detectable in 6 h or 8 h, but with added ALA already after 4 h or 6 h. The penetration of BUD from the PG/water medium after the addition of ALA increased by 480% within 8 h and on the average by 210% within 20-24 h. The penetration of BUD from the buffer after the addition of ALA was on the average by 170 % higher within 20-24 h. The permeation of BUD from isopropyl myristate after the addition of ALA increased on the average by 30% within 6-8 h and by 50% within 20-24 h.

Fig. 17: Comparison of the penetration of dexamethasone (DEX) through the skin from various media without and with the presence of 0.1% (in relation to DEX amount) of micronized or nanonized alaptide (ALA, NALA) in time: DEX from the propylene glycol (PG)/water (1:1) medium and from buffer (pH 7.4). Without the addition of ALA DEX (10 mg/mL, 100 %) penetrated from neither of the media. After the addition of ALA DEX was detected within 3 h from both media, and after the addition of NALA DEX significantly penetrated within 30 min. Fig. 18: Comparison of the penetration of dexamethasone acetate (DEA) through the skin from various media without and with the presence of 0.1% (in relation to DEA amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (IPM). Without the addition of ALA DEA (10 mg/mL, 100 %) was detected after 6 h or after 20 h, but ALA already after 4 h or 6 h. The addition of ALA increased the penetration of DEA from the PG/water medium on the average by 110% within 6-8 h and by 30% within 20-24 h. After the addition of ALA DEA penetrated from the buffer on the average by 190% more within 20-24 h. The addition of ALA increased the penetration of DEAfrom isopropyl myristate on the average by 20% within 20-24 h.

Fig. 19: Comparison of the penetration of fluocinolone (FLC) through the skin from various media without and with the presence of 0.1% (in relation to FLC amount) of micronized or nanonized alaptide (ALA, NALA) in time: from the propylene glycol (PG)/water (1:1) medium and from buffer (pH 7.4). Without the addition of ALA, FLC (10 mg/mL, 100%) penetrated from neither of the media. After the addition of ALA FLC was not found from the buffer, but from the PG/water medium it was found within 30 min, and after the addition of NALA FLC significantly penetrated from both media within 30 min, in the case of the PG/water medium on the average by 1100% within 1 h.

Fig. 20: Comparison of the penetration of fluocinolone-acetonide (FLA) through the skin from various media without and with the presence of 0.1% (in relation to FLA amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (IPM). Without the addition of ALA FLA (10 mg/mL, 100%) penetrated after 4 h or 8 h, from the media with ALA already after 4 h. The penetration of FLA from the PG/water medium after the addition of ALA increased on the average by 180% within 4-8 h and by 130% within 20-24 h. With the addition of ALA FLA penetrated from the buffer by 250% more within 8 h and on the average by 170 % more within 20-24 h. The addition of ALA increased the permeation of FLA from isopropyl myristate on the average by 590% within 4 h, on the average by 270% within 6-8 h and by 30% within 20-24 h. Fig. 21: Comparison of the penetration of hydrocortisone acetate (HKA) through the skin from various media without and with the presence of 0.1% (in relation to HKA amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (IPM). Without the addition of ALA HKA (10 mg/mL, 100%) penetrated after 20 h or did not penetrate at all, but from the media with ALA already after 4 h or after 20 h. The penetration of HKA from the buffer and from isopropyl myristate after the addition of ALA increased on the average by 40% within 20-24 h.

Fig. 22: Comparison of the penetration of prednisolone acetate (PLA) through the skin from various media without and with the presence of 0.1% (in relation to PLA amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (IPM). Without the addition of ALA PLA (10 mg/mL, 100%) penetrated after 4 h or 20 h, but from the media with ALA already after 4 h or after 6 h. The penetration of PLA from the PG/water medium after the addition of ALA increased on the average by 300% within 20-24 h. With ALA PLA penetrated from the buffer on the average by 270% more within 4-8 h and by 180% more within 20-24 h. The permeation of PLA from isopropyl myristate after the addition of ALA increased on the average by 30% within 20-24 h.

Fig. 23: Comparison of the penetration of prednisolone (PDL) through the skin from various media without and with the presence of 0.1% (in relation to PDL amount) of micronized or nanonized alaptide (ALA, NALA) in time: from the propylene glycol (PG)/water (1:1) medium and from buffer (pH 7.4). Without the addition of ALA PDL (10 mg/mL, 100%) penetrated from neither of the media, but after the addition of ALA PDL permeated from the PG/water medium within 90 min. After the addition of NALA the permeation of PDL from both media significantly increased within 30 min, in case of the PG/water medium on the average by 275% within 3 h.

Fig. 24: Comparison of the penetration of prednisone (PDN) through the skin from various media without and with the presence of 0.1% (in relation to PDN amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (IPM). Without the addition of ALA PDN (lO mg/ml, 100%) penetrated after 4 h from all the media, nevertheless the addition of ALA increased the penetration of PDN from the PG/water medium by 680% within 4 h, by 370% within 6-8 h and by 110% within 20-24 h. With ALA PDN penetrated from the buffer by 650% more within 4 h, on the average by 370 % more within 6-8 h and by 150% more within 20-24 h. The addition of ALA increased the permeation of PDN from isopropyl myristate on the average by 320% within 4 h, on the average by 200% within 6-8 h and by 90% within 20-24 h.

Fig. 25: Comparison of the penetration of triamcinolone (TRO) through the skin from various media without and with the presence of 0.1% (in relation to TRO amount) of micronized or nanonized alaptide (ALA, NALA) in time: from the propylene glycol (PG)/water (1:1) medium and from buffer (pH 7.4). The addition of ALA to the buffer increased the penetration of TRO (10 mg/ml, 100%) by 174% within 30 min, and the addition of ALA to the PG/water medium increased the penetration of TRO by 267% within 30 min.

Fig. 26: Comparison of the penetration of triamcinolone acetonide (TRA) through the skin from various media without and with the presence of 0.1% (in relation to TRA amount) of micronized or nanonized alaptide (ALA, NALA) in time: from the propylene glycol (PG)/water (1:1) medium and from buffer (pH 7.4). Without the addition of ALA TRA (10 mg/mL, 100 %) penetrated from both media after 90 min or 120 min. After the addition of ALA to the buffer the penetration of TRA increased by 5% within 90 min. The addition of NALA to the PG/water medium caused the penetration of TRA already in 30 min. The addition of ALA to the PG/water medium increased the penetration of TRA in comparison with NALA by 790% in 30 min.

Fig. 27: Comparison of the penetration of dexamethasone acetate (DEA) through the skin from gel without and with the presence of 0.1% (in relation to DEA amount) of micronized alaptide (ALA) in time: DEA penetrated from the formulation without and with the addition of ALA after 20 h, nevertheless the addition of ALA increased the penetration of DEA on the average by 10%.

Fig. 28: Comparison of the penetration of fluocinolone acetonide (FLA) through the skin from hydro-cream, oleo-cream and ointment without and with the presence of 0.1% (in relation to FLA amount) of micronized alaptide (ALA) in time: the highest penetration of FLA was observed from the oleo-cream, the penetration from the hydro-cream was on the average by 14% less and from the ointment by almost 96% less. The penetration of FLA from the hydro-cream with added ALA increased on the average by 260% within 4-8 h and by 110% within 20-24 h. The penetration of FLA from the oleo-cream with added ALA increased on the average by 230% within 4-8 h and by 160% within 20-24 h. The penetration of FLA from the ointment with added ALA increased on the average by 20% within 20-24 h.

Fig. 29: Comparison of the penetration of triamcinolone acetonide (TRA) through the skin from oleo-ointment without and with the presence of 0.1% (in relation to TRA amount) of micronized or nanonized alaptide (ALA, NALA) in time. TRA from the formulation without ALA/NALA was not found within 3 h. After the addition of ALA TRA was found already within 30 min, and the addition of NALA increased the penetration of TRA by 273%.

Fig. 30: Comparison of the penetration of triamcinolone acetonide (TRA) through the skin from hydro-cream without and with the presence of 0.1% (in relation to TRA amount) of micronized or nanonized alaptide (ALA, NALA) in time. TRA from the formulation without ALA/NALA was not found within 3 h. After the addition of ALA TRA was found already after 90 min. After the addition of NALA TRA was fou d within 1 h, and within 90 miri the penetration of -TRA with the addition of NALA increased by 124% in comparison with ALA.

Fig. 31: Comparison of the penetration of triamcinolone acetonide (TRA) through the skin from methylcellulose gel without and with the presence of 0.1% (in relation to TRA amount) of micronized or nanonized alaptide (ALA, NALA) in time. TRA from the formulation without ALA/NALA was not found within 3 h. After the addition of ALA TRA was found already after 120 min. The penetration of TRA with the addition of NALA was increased by 228% in comparison with ALA.

Fig. 32: Comparison of the penetration of triamcinolone acetonide (TRA) through the skin from carbomer gel without and with the presence of 0.1% (in relation to TRA amount) of micronized or nanonized alaptide (ALA, NALA) in time. TRA from formulation without ALA/NALA was not found within 3 h. After the addition of ALA TRA was found already within 120 min. The addition of NALA increased the penetration of TRA by 5% in comparison with ALA.

Fig. 33: Comparison of the penetration of amoxicillin (AMX) through the skin from various media without and with the presence of 0.1% (in relation to AMX amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (IPM). The penetration of AMX (10 mg/mL, 100%) from the PG/water medium after the addition of ALA increased by 55% within 8 h and on the average by 92% without 20-24 h. The addition of ALA increased the penetration of AMX from the buffer on the average by 106% within 8 h. The penetration of AMX from isopropyl myristate with ALA increased on the average by 5% within 8 h and by 30% within 20-24 h.

Fig. 34: Comparison of the penetration of ampicillin (AMP) through the skin from various media without and with the presence of 0.1% (in relation to AMP amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (ΓΡΜ). The penetration of AMP (10 mg/mL, 100%) from the PG/water medium after the addition of ALA was increased by 145% within 8 h. The addition of ALA increased the penetration of AMP from the buffer on the average by 35% within 8 h. The addition of ALA increased the penetration of AMP from isopropyl myristate on the average by 54% within 8 h.

Fig. 35: Comparison of the penetration of oxacillin (OXL) through the skin from various- media without and with the presence of 0.1% (in relation to OXL amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (ΓΡΜ). The penetration of OXL (10 mg/mL, 100%) from the PG/water medium after the addition of ALA increased by 150% within 8 h and on the average by 80% within 20-24 h. The addition of ALA increased the penetration of OXL from the buffer on the average by 27% within 24 h. The addition of ALA increased the penetration of OXL from isopropyl myristate on the average by 177% within 8 h.

Fig. 36: Comparison of the penetration of benzylpenicillin (penicillin G, PEG) through the skin from various media without and with the presence of 0.1% (in relation to PEG amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (ΓΡΜ). The penetration of PEG (10 mg/mL, 100%) from the PG/water medium after the addition of ALA was increased by 136% within 8 h. The addition of ALA increased the penetration of PEG from the buffer on the average by 16% within 24 h and from isopropyl myristate on the average by 5% within 20-24 h. Fig. 37: Comparison of the penetration of phenoxymethylpenicillin (penicillin V, PEV) through the skin from various media without and with the presence of 0.1% (in relation to PEV amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium, from buffer (pH 7.4) and from isopropyl myristate (IPM). The penetration of PEV (10 mg/mL, 100%) from the PG/water medium after the addition of ALA increased by 56% within 8 h and on the average by 45% within 20-24 h. The addition of ALA increased the penetration of PEV from the buffer on the average by 43% within 8 h and from isopropyl myristate on the average by 34% within 8 h.

Fig. 38: Comparison of the penetration of ofloxacin (OFX) through the skin from various media without and with the presence of 0.1% (in relation to OFX amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water'(l:l) medium, from buffer (pH 7.4) and from isopropyl myristate (IPM). The penetration of OFX (10 mg/mL, 100%) from the PG/water medium after the addition of ALA was increased by 54% within 8 h. The addition of ALA increased the penetration of OFX from the buffer on the average by 137% within 8 h and by 85% within 20-24 h and from isopropyl myristate on the average by 36% within 8 h.

Fig. 39: Comparison of the penetration of ofloxacin (OFX) through the skin from methylcellulose and carbomer gel without and with the presence of 0.1% (in relation to OFX amount) of micronized alaptide (ALA) in time. The penetration of OFX from the methylcellulose gel after the addition of ALA increased on the average by 1040% within 4-8 h and by 136% within 20-24 h. The addition of ALA increased the penetration of OFX from the carbomer gel on the average by 200% within 4-8 h and by 80 % within 20-24 h.

Fig. 40: Comparison of the penetration of sulfafhiazole (SFT) through the skin from oleo- ointment without and with the presence of 0.1% (in relation to SFT amount) of micronized or nanonized alaptide (ALA, NALA) in time. The penetration of SFT from the formulation after the addition of ALA increased on the average by 10% within 8 h and by 37% within 12-24 h. The addition of NALA increased the penetration of SFT on the average by 180% within 8 h and by 330% within 12-^24 h.

Fig. 41: Comparison of the penetration of chloramphenicol (CRF) through the skin from various media without and with the presence of 0.1% (in relation to CRF amount) of micronized alaptide (ALA) in time: from the propylene glycol (PG)/water (1:1) medium and from buffer (pH 7.4). After the addition of ALA the penetration of CRF (10 mg/mL, 100%) from the PG/water medium was increased by 7% within 24 h and from the buffer by 1% within 24 h.

Fig. 42: Comparison of the penetration of chloramphenicol (CRF) through the skin from oleo-ointment without and with the presence of 0.1% (in relation to CRF amount) of micronized or nanonized alaptide (ALA, NALA) in time. The penetration of CRF from the formulation after the addition of ALA increased on the average by 90% within 8 h. The penetration of CRF from the formulation after the addition of NALA increased on the average by 360% within 8 h and by 1030% within 24 h.

Fig. 43: Comparison of the penetration of neomycin sulphate (NMC) through the skin from the propylene glycol (PG)/water (1:1) medium without and with the presence of 0.1% (in relation to NMC amount) of micronized alaptide (ALA). The addition of ALA increased the penetration of NMC (10 mg/mL, 100%) from the PG/water medium by 165% within 30 min.

Fig. 44: Comparison of the penetration of mupirocin (MPC) through the skin from hydro- ointment without and with the presence of 0.1% (in relation to MPC amount) of micronized or nanonized alaptide (ALA, NALA) in time. After the addition of ALA the penetration of MPC (10 mg/mL, 100 %) from the formulation increased on the average by 144% within 30 min and almost by 400% within 60 min; thus the penetration of MPC through the skin was significantly increased by ALA. Conversely, the addition of NALA significantly inhibited (decreased by 60% within the 1st hour) the penetration of MPC through the skin, so MPC acted only on the skin surface.

Fig. 45: Comparison of the penetration of pyrazinamide (PZA) through the skin from buffer (pH 7.4) without and with the presence of 0.1% (in relation to PZA amount) of micronized or nanonized alaptide (ALA, NALA) in time. The addition of ALA increased the penetration of PZA (10 mg/mL, 100 %) by 63% within 8 h and by 303% within 24 h. The addition of NALA increased the penetration of PZA by 125% within 24 h.

Fig. 46: Comparison of the penetration of pyrazinamide (PZA) through the skin from carbomer gel without and with the presence of 0.1% (in relation to PZA amount) of micronized or nanonized alaptide (ALA, NALA) in time. The addition of ALA increased the penetration of PZA (10 mg/mL, 100 %) from the gel by 16% within 24 h, and the addition of NALA to the gel increased the penetration of PZA on the average by 16% within 24 h.

Fig. 47: Comparison of the penetration of fluconazole (FLU) through the skin from the propylene glycol (PG)/water (1:1) medium without and with the presence of 0.1% (in relation to FLU amount) of micronized or nanonized alaptide (ALA, NALA) in time. The penetration of FLU (10 mg/mL, 100 %) from PG/water medium after the addition of NALA was increased by 59% within 24 h.

Fig. 48: Comparison of the penetration of fluconazole (FLU) through the skin from oleo- ointment and hydro-cream without and with the presence of 0.1% (in relation to FLU amount) of nanonized alaptide (NALA) in time. The penetration of FLU from the ointment after the addition of NALA increased on the average by 150 % within 8 h and by 350% within 12-24 h. The addition of NALA increased FLU penetration from the cream by 28% within 12-24 h.

Fig. 49: Comparison of the penetration of aciclovir (ACL) through the skin from various media without and with the presence of 0.1% (in relation to ACL amount) of micronized or nanonized alaptide (ALA, NALA) in time: from the propylene glycol (PG)/watef (1:1) medium and from buffer (pH 7.4). After the addition of ALA the penetration of ACL (10 mg/mL, 100 %) from the PG/water medium increased by 114% already within 30 in. The addition of NALA to the PG/water medium increased the penetration of ACL by 158% within 30 min and by 280% within 2 h. The addition of ALA to the buffer increased the penetration of ACL by 126% within 30 min and by 440% within 2 h.

Fig. 50: Comparison of the penetration of aciclovir (ACL) through the skin from hydro- cream or carbomer gel without and with the presence of 0.1% (in relation to ACL amount) of micronized or nanonized alaptide (ALA, NALA) in time. The addition of NALA to the hydro-cream increased the penetration of ACL (10 mg/mL, 100 %) by 25% within 30 min, and the addition of ALA to the gel increased the penetration of ACL by 37% within 30 min. WORKING EXAMPLES:

Example 1 Preparation of nanonized alaptide. The suspension of micronized alaptide (30 g), polyvinylpyrrolidone (30 g) and purified water (240 mL, during milling was diluted by addition of additional 150 mL) was initially mixed for 12 h at laboratory temperature and then filtered through a mill sieve. The milling procedure was performed using a nanomill NETZSCH (Germany) with glass beads (0.3 mm); the rotor speed was 986 rpm; the pump speed was 30 rpm; the temperature in the grinding chamber was within 17-20 °C. The rotor speed was increased to 1500 rpm after 6 h of milling. The total time of milling was 57.5 h. The content of alaptide in the suspension was 38.76 g/L (determined by RP-HPLC). The particle size of the prepared nanonized alaptide was measured by Sympatec NANOPHOX 0138 P (Germany), and the particle size X5o-X9o was up to 900 nm.

Example 2 In vitro transdermal penetration experiments performed using Franz diffusion cell. Full thickness dorsal skin from porcine ear was cut into pieces and stored at -20 °C until needed. Skin samples were slowly thawed (at 4 °C overnight and then at ambient temperature) prior to each experiment. Prior to the experiment, the skin was kept in contact with the receptor phase for 0.5 h at 37+0.5 °C. To the donor part of a Franz diffusion cell with the volume of 1 mL and the surface area of 63.585 mm an investigated sample was applied in form of solution, suspension, emulsion, gel, cream or ointment, always with the drug concentration of 10 mg/mL. As a sample the drug itself (active pharmaceutical ingredient) was used or the drug in formulation with various concentrations of micronized or nanonized alaptide (nanonized alaptide in the amount corresponding to the concentration of micronized alaptide) from 0.001 to 10 g, otherwise from 0.01 to 100% (w/w in relation to the drug amount). The skin was mounted with the epidermal side up between the donor and the receptor compartments of the diffusion cell. The receptor compartment (volume 5.2 mL) was filled with phosphate buffered saline (pH 7.4), with a mixture of propylene glycol:water (1:1) or with isopropyl myristate and maintained at 37+0.5 °C using a circulating water bath. The receptor compartment was continuously stirred using a magnetic stirring bar (800 rpm). Than a sample was applied to the skin surface, and the donor compartment of the cell was covered by Parafilm®. Samples (0.5 mL) were taken from the receptor phase in time intervals, and the cell was refilled with an equivalent volume of the fresh buffer, the mixture of propylene glycol:water (1:1) or isopropyl myristate solution. For each compound, a minimum of three determinations were performed using skin fragments from a minimum of 2 animals. The penetration through the skin of the drug alone or the drug in formulation without alaptide (as control samples) was monitored, then the penetration of the drug with various concentrations of micronized or nanonized alaptide (nanonized alaptide in the amount corresponding to the concentration of micronized alaptide) was monitored. Also the penetration of the drug from the particular pharmaceutical composition (ointment, cream, gel) with various concentrations of micronized or nanonized alaptide (nanonized alaptide in the amount corresponding to the concentration of micronized alaptide) was monitored. The concentration of the penetrated drug was determined by RP-HPLC method with UV-VIS detection. All results of the all investigated samples (non-steroidal anti-inflammatory drugs and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics) are summarized in the presented figures with tables.

Example 3 100 g of gel containing from 0.001 to 10 g, otherwise from 0.01 to 100% (w/w irt relation to the drug amount) of micronized or nanonized alaptide (in the amount corresponding to the concentration of micronized alaptide) an a drug (ibuprofen, diclofenac, meloxicam, nimesulide, dexamethasone acetate, ofloxacin, pyrazinamide, aciclovir) was prepared, and permeation experiments were performed according to Example 2. -

Example 4 100 g of cream containing from 0.001 to 10 g, otherwise from 0.01 to 100% (w/w in relation to the drug amount) of micronized alaptide or nanonized alaptide (in the amount corresponding t the concentration of micronized alaptide) and a drug (ibuprofen, diclofenac, meloxicam, nimesulide, fluocinolone acetonide, fluconazole, aciclovir) was prepared, and permeation experiments were performed according to Example 2.

Example 5 100 g of ointment containing from 0.001 to 10 g, otherwise from 0.01 to 100% (w/w in relation to the drug amount) of micronized alaptide or nanonized alaptide (in the amount corresponding to the concentration of micronized alaptide) and a drug (ibuprofen, diclofenac, meloxicam, nimesulide, fluocinolone acetonide, sulfathiazole, chloramphenicol, mupirocin, fluconazole, aciclovir) was prepared, and permeation experiments were performed according to Example 2. CLAIMS:

1. The utilization of alaptide, structural formula /, as a chemical modifier of transdermal penetration of non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics sampled from the classes of antibacterials/antimycobacterials, antimycotics and antivirotics in the pharmaceutical composition suitable for transdermal application.

2. The utilization of alaptide according to Claim 1 characterized by the fact that the used non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) are sampled from the following list: phenylbutazone, oxyphenbutazone, kebuzoiie, tribuzone, sulfinpyrazone, azapropazone, mofebutazone, clofezon, suxibuzone, flufenamic acid, niflumic acid, mefenamic acid, tolfenamic acid, meclofenamic acid, aclantate, etofenamate, flunixine, ibufenac, alclofenac, diclofenac, indometacin, acemetacin, tropesin, sulindac, lonazolac, tolmetin, ketorolac, nabumetone, fenbumetone, indobufene, zomepirac, bumadizone, etodolac, fentiazac, difenpiramide, oxametacin, proglumetacin, aceklofenac, bufexamac, felbinac, bendazac, fentiazac, nifenazone, ibuprofen, naproxen, ketoprofen, suprofen, flurbiprofen, pirprofen, flobufen, fenoprofen, fenbufen, benoxaprofen, indoprofen, oxaprozin, ibuproxam, dexibuprofen, flunoxaprofen, alminoprofen, dexketoprofen, vedaprofen, carprofen, tepoxalin, tiaprofenic acid, tenoxicam, droxicam, lornoxicam, piroxicam, meloxicam, isoxicam, nimesulide, celecoxib, rofecoxib, valdecoxib, parecoxib, etoricoxib, lumiracoxib, firocoxib, robenacoxib, mavacoxib, cimicoxib, prochazon, orgotein, feprazone, diacerein, morniflumate, tenidap, oxaceprol, glucosamine, benzydamine, glycosaminoglycan polysulfate, chondroitin sulfate, pentosan polysulfate, aminopropionitrile; thus NSAIDs from the classes of salicylates, pyrrazolidindipnes, fenamates, derivatives of arylalkanoic acids (fenacs, profens), oxicames, coxibes and other non-steroidal compounds, which mechanism of action is connected with inhibition of the metabolic pathway of arachidonic acid and generation of pro-inflammatory substances such as prostacyclins, prostaglandins, thromboxanes and leukotrienes. 3. The utilization of alaptide according to Claim 1 characterized by the fact that the used antipyretics/non-opiate analgesics are sampled from the following list: paracetamol, bucetin, propacetamol, salicylic acid, acetylsalicylic acid, choline salicylate, lysine salicylate, salicylate sodium or potassium, imidazole salicylate, morpholine salicylate, salsalate, ethenzamide, guacetisal, carbasalate calcium, salicylamide, methylsalicylate, aloxiprin, lysine acetylsalicylate, benorilate, gentisic acid, dipyrocetyl, diflunisal, phenazone, aminophenazone, propyphenazone, ramiphenazone, metamizole, rimazoline, glafenine, floctafenine, viminol, nefopam, flupirtine, ziconotide, menthol, nabiximols, other antipyretics/non-opiate analgesics from the classes of anilines, salicylic acid derivatives, pyrazolones and other compounds affecting body thermoregulation or belonging to non-opiate analgesics.

4. The utilization of alaptide according to Claim 1 characterized by the fact that the used glucocorticoids are sampled from the following list: hydrocortisone, hydrocortisone acetate, fludrocortisone, tixocortol, medrysone, dexamethasone, dexamethasone acetate, prednisone, prednisolone, prednisolone acetate, fluprednisolone, mefhylprednisolone, methylprednisolone aceponate, fluorometholone, difluprednate, mazipredone, betamethasone, betamethasone dipropionate, paramethasone, flumethasone, desoximetasone, fluocortolone, diflucortolone, clocortolone, prednylidene, fluprednidene, triamcinolone, triamcinolon acetonide, flunisolide, desonide, prednicarbate, budesonide, fluocinolone acetonide, fluocinonide, ciclesonide, mometasone furoate, clobetasol propionate, other glucocorticoids from the classes of hydrocortisone, prednisolone, dexamethasone, triamcinolone and other steroid compounds (steroidal antirinflammatory drugs), which mechanism of action is connected with interference to the metabolism of arachidonic acid and with following inhibition of generation of pro-inflammatory icosanoids (prostacyclins, prostaglandins, thromboxanes, leukotrienes) and/or a decrease in production of cytokines and/or a decrease in histamine liberation.

5. The utilization of alaptide according to Claim 1 characterized by the fact that the used antibiotics and/or antibacterial/antimycobacterial chemotherapeutics are sampled from the following list: beta-lactam antibiotics (penicillins, carbapenems, monobactams and/or cephalosporins, carbacephems, oxacephems), macrolides, tetracyclines, aminoglycosides, polypeptides, glycopeptides, lincosamides, lipopeptides, ansamycins, fiisidic acid, linezolid, classes of sulfonamides, quinolones, amphenicols, nitrofurans, nitroimidazoles, ^-aminosalicylic acid, cycloserine, isoniazid, pyrazinamide, ethionamide, protionamide, ethambutol, clofazimine, dapsone as well as other natural, semisynthetic or synthetic antibacterially effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of bacterial pathogens and causes their death.

6. The utilization of alaptide according to Claim 1 characterized by the fact that the used antimycotics are sampled from the following list: polyenes, griseofulvin, imidazoles, triazoles, allylamines and other non-azole ergosterol biosynthesis inhibitors, thiocarbamates, glucan synthesis inhibitors (echinocandines, pneumocandines, papulacandines), antimetabolites (flucytosine), ciclopirox, amorolfine and other antifungal effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of fungal pathogens and causes their

death. . ·. · .

7. The utilization of alaptide according to Claim 1 characterized by the fact that the used antivirotics are sampled from the following list: pyrimidine and purine nucleotides, reverse transcriptase inhibitors, HIV-protease inhibitors, neuramidase inhibitors, amantadine, interferons and foscarnet as well as other antiviral effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of viral pathogens and causes their death.

8. Pharmaceutical compositions for transdermal application containing non-steroidal anti inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) and/or antipyretics/non-opiate analgesics and/or glucocorticoids and/or antimicrobial chemotherapeutics sampled from the classes of antibacterials/antimycobacterials, antimycotics, antivirotics and simultaneously containing alaptide as a chemical transdermal penetration modifier. '

9. Pharmaceutical compositions according to Claim 8 characterized by the content of alaptide from 0.01 to 100% by weight in relation to the amount of the used drug (active pharmaceutical ingredient).

10. Pharmaceutical compositions according to Claims 8 and 9 characterized by the content of micronized alaptide or alaptide in the form of nanoparticles. 11. Pharmaceutical compositions according to Claims 8 to 10 characterized by the fact that the pharmaceutical formulation is in the form of oleo-ointment, hydro-ointment, oleo- cream, hydro-cream or gel. .

12. Pharmaceutical compositions according to Claims 8 to 1 1 characterized by the fact that the used non-steroidal anti-inflammatory drugs (otherwise non-steroidal antiphlogistics or NSAIDs) are sampled from the following list: phenylbutazone, oxyphenbutazone, kebuzone, tribuzone, sulfinpyrazone, azapropazone, mofebutazone, clofezon, suxibuzone, flufenamic acid, niflumic acid, mefenamic acid, tolfenamic acid, meclofenamic acid, aclantate, etofenamate, flunixine, ibufenac, alclofenac, diclofenac, indometacin, acemetacin, tropesin, sulindac, lonazolac, tolmetin, ketorolac, nabumetone, fenbumetone, indobiifene, zomepirac, bumadizone, etodolac, fentiazac, difenpiramide, oxametacin, proglumetacin, aceklofenac, bufexamac, felbinac, bendazac, fentiazac, nifenazone, ibuprofen, naproxen, ketoprofen, suprofen, flurbiprofen, pirprofen, flobufen, fenoprofen, fenbufen, benoxaprofen, indoprofen, oxaprozin, ibuproxam, dexibuprofen, flunoxaprofen, alminoprofen, dexketoprofen, vedaprofen, carprofen, tepoxalin, tiaprofenic acid, tenoxicam, droxicam, lornoxicam, piroxicam, meloxicam, isoxicam, nimesulide, celecoxib, rofecoxib, valdecoxib, parecoxib, etoricoxib, lumiracoxib, firocoxib, robenacoxib, mavacoxib, cimicoxib, prochazon, orgotein, feprazone, diacerein, morniflumate, tenidap, oxaceprol, glucosamine, benzydamine, glycosaminoglycan polysulfate, chondroitin sulfate, pentosan polysulfate, aminopropionitrile; thus NSAIDs from the classes of salicylates, pyrrazolidindiones, fenamates, derivatives of arylalkanoic acids (fenacs, profens), oxicames, coxibes and other non-steroidal compounds, which mechanism of action is connected with inhibition of the metabolic pathway of arachidonic acid and generation of pro-inflammatory substances such as prostacyclins, prostaglandins, thromboxanes and leukotrienes.

13. Pharmaceutical compositions according to Claims 8 to 11 characterized by the fact that the used antipyretics/non-opiate analgesics are sampled from the following list: paracetamol, bucetin, propacetamol, salicylic acid, acetylsalicylic acid, choline salicylate, lysine salicylate, salicylate sodium or potassium, imidazole salicylate, morpholine salicylate, salsalate, ethenzamide, guacetisal, carbasalate calcium, salicylamide, methylsalicylate, aloxiprin, lysine acetylsalicylate, benorilate, gentisic acid, dipyrocetyl, diflunisal, phenazone, aminophenazone, propyphenazone, ramiphenazone, metamizole, rimazoline, glafenine, floctafenine, viminol, nefopam, flupirtine, ziconotide, menthol, nabiximols, other antipyretics/non-opiate analgesics from the classes of anilines, salicylic acid derivatives, pyrazolones and other compounds affecting body thermoregulation or belonging to non-opiate analgesics.

14. Pharmaceutical compositions according to Claims 8 to 11 characterized by the fact that the used glucocorticoids are sampled from the following list: hydrocortisone, hydrocortisone acetate, fludrocortisone, tixocortol, medrysone, dexamefhasone, dexamethasone acetate, prednisone, prednisolone, prednisolone acetate, fluprednisolone, methylprednisolone, methylprednisolone aceponate, fluorometholone, difluprednate, mazipredone, betamethasone, betamethasone dipropionate, paramethasone, flumethasone, desoximetasone, fluocortolone, diflucortolone, clocortolone, prednylidene, fluprednidene, triamcinolone, triamcinolon acetonide, flunisolide, desonide, prednicarbate, budesonide, fluocinolone acetonide, fluocinonide, cielesonide, mometasone furoate, clobetasol propionate, other glucocorticoids from the classes of hydrocortisone, prednisolone, dexamethasone, triamcinolone and other steroid compounds (steroidal anti-inflammatory drugs), which mechanism of action is connected with interference to the metabolism of arachidonic acid and with following inhibition of generation of pro-inflammatory icosanoids (prostacyclins, prostaglandins, thromboxanes, leukotrienes) and/or a decrease in production of cytokines and/or a decrease in histamine liberation.

15. Pharmaceutical compositions according to Claims 8 to 11 characterized by the fact that the used antibiotics and/or antibacterial/antimycobacterial chemotherapeutics are sampled from the following list: beta-lactam antibiotics (penicillins, carbapenems, monobactams and/or cephalosporins, carbacephems, oxacephems), macrolides, tetracyclines, aminoglycosides, polypeptides, glycopeptides, lincosamides, lipopeptides, ansamycins, fusidic acid, linezolid, classes of sulfonamides, quinolones, amphenicols, nitrofurans, nitroimidazoles, /^-aminosalicylic acid, cycloserine, isoniazid, pyrazinamide, ethionamide, protionamide, ethambutol, clofazimine, dapsone as well as other natural, semisynthetic or synthetic antibacterially effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of bacterial pathogens and causes their death.

16. Pharmaceutical compositions according to Claims 8 to 11 characterized by the fact that the used antimycotics are sampled from the following list: polyenes, griseofulvin, imidazoles, triazoles, allylamines and other non-azole ergosterol biosynthesis inhibitors, thiocarbamates, glucan synthesis inhibitors (echinocandines, pneumocandines, papulacandines), antimetabolites (flucytosine), ciclopirox, amorolfine and other antifungal effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of fungal pathogens and causes their death.

Pharmaceutical compositions according to Claims 8 to 11 characterized by the fact that the used antivirotics are sampled from the following list: pyrimidine and purine nucleotides, reverse transcriptase inhibitors, HIV-protease inhibitors, neuramidase inhibitors, amantadine, interferons and foscarnet as well as other antiviral effective compounds, which mechanism of action is connected with inhibition of the growth and/or multiplication of viral pathogens and causes their death.

A . CLASSIFICATION O F SUBJECT MATTER INV. A61K9/06 A61K9/14 A61K47/10 A61K47/18 A61K47/32 A61K47/38 A61K9/00 ADD. According to International Patent Classification (IPC) or to both national classification and IPC

B . FIELDS SEARCHED Minimum documentation searched (classification system followed by classification symbols) A61K

Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched

Electronic data base consulted during the international search (name of data base and, where practicable, search terms used)

EPO-Internal , BIOSIS, WPI Data

C . DOCUMENTS CONSIDERED TO B E RELEVANT

Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.

JULINEK 0 ET AL: " Product of al apti de 1-17 synthesi s: Determi nati on of the absol ute confi gurati on" , JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, NEW YORK, NY, US, vol . 53 , no. 4, 1 December 2010 (2010-12-01) , pages 958-961 , XP027208247 , ISSN : 0731-7085 [retri eved on 2010-08-01] the whol e document -/-

X| Further documents are listed in the continuation of Box C . □ See patent family annex. * Special categories of cited documents : "T" later document published after the international filing date or priority date and not in conflict with the application but cited to understand "A" document defining the general state of the art which is not considered the principle or theory underlying the invention to be of particular relevance "E" earlier application or patent but published o n or after the international "X" document of particular relevance; the claimed invention cannot be filing date considered novel or cannot be considered to involve an inventive "L" documentwhich may throw doubts on priority claim(s) orwhich is step when the document is taken alone cited to establish the publication date of another citation or other "Y" document of particular relevance; the claimed invention cannot be special reason (as specified) considered to involve an inventive step when the document is "O" document referring to an oral disclosure, use, exhibition or other combined with one o r more other such documents, such combination means being obvious to a person skilled in the art "P" document published prior to the international filing date but later than the priority date claimed "&" document member of the same patent family

Date of the actual completion of the international search Date of mailing of the international search report

16 October 2012 31/10/2012

Name and mailing address of the ISA/ Authorized officer European Patent Office, P.B. 5818 Patentlaan 2 NL - 2280 HV Rijswijk Tel. (+31-70) 340-2040, Fax: (+31-70) 340-3016 Gimenez Mi ral l es , J C(Continuation). DOCUMENTS CONSIDERED TO BE RELEVANT

Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.

DATABASE MEDLINE [Onl i ne] 1-17 US NATIONAL LIBRARY OF MEDICINE (NLM) , BETHESDA, MD, US; June 2011 (2011-06) , VITKOVA Z ET AL: " [Infl uence of membranes on al apti de permeati on from hydrogel s] . " , XP002685293 , Database accessi on no. NLM21838143 c b s c c & VITKOVA Z ET AL: " [Infl uence of membranes on al apti de permeati on from hydrogel s] . " , CESKA A SLOVENSKA FARMACI E : CASOPIS CESKE FARMACEUTICKE SPOLECNOSTI A SLOVENSKE FARMACEUTICKE SPOLECNOSTI JUN 2011 LNKD- PUBMED:21838143 , vol . 60, no. 3 , June 2011 (2011-06) , pages 132-136, ISSN : 1210-7816