I|||||III US005567716A United States Patent 19 11 Patent Number: 5,567,716 Della Valle et al. (45) Date of Patent: Oct. 22, 1996

54 TRANS AND CISTRAUMATIC ACID SALTS Chemical Abstracts, vol. 83, No. 3, Abstract No. 29, 1165, p. HAVING CICATRIZANT ACTIVITY 40, Jul 21, 1975. ASSOCIATED TO BACTERIOSTATIC, Nakayama et al, Bull Chem. Soc. Jpn. vol. 64, pp. 358-365 ANTIVIRAL, ANTIBIOTIC OR ANTIFUNGAL (1991). ACTIVITY Breslow et al., J. Am. Chem. Soc., vol. 103 (10) pp. 2905-2907 (1981). 75) Inventors: Francesco Della Valle; Silvana Zimmerman etal, Plant Physiol. (1979) 63,536-541, "Iden Lorenzi, both of Padova, Gabriele tification of Traumatin, A Wound Hormone, as Marcolongo, Carrara San Giorgio, all 12-oxo-Trans-10 Dodecenoic Acid'. of Italy English et al., J. Biol. Chem. vol. 121, No. 2, (1937), 791-799, "The Wound Hormones of Plants: I. Traumatin, 73 Assignee: Lifegroup S.p.A., Rome, Italy the Active Principle of the Bean Test'. Goldemberg, J. Soc. Cosmet. Chem., 28, (Nov. 1977), 21 Appl. No.: 155,153 667-679, “Reduction of Topical Irritation”. Miyamoto et al, J. Soc. Cosmet, Chem. Japan, vol. 22, No. 22 Filed: Nov. 19, 1993 4, (1989), 254-262, "Effects of Cosmetics Containing Bio 30 Foreign Application Priority Data active Substances on Skin'. Rodriguez-Bigas et al, Plastic and Reconstructive Surgery, Nov. 23, 1992 IT Italy ...... MI92A2674 vol. 81, No. 3 (Mar. 1988), 386-389, “Comparative Evalu (51) Int. Cl." ...... A61K 31/195; A61K 31/20 ation of Aloe Vera in the Management of Burn Wounds in 52 U.S. Cl...... 514/332; 534/676; 534/773; Guinea Pigs'. 544/242; 548/161; 548/164; 548/310.1; Phytomorphology, vol. 25, No. 3, 1975, pp. 265-261 (sic), 548/341.5; 54.6/105; 546/267; 546/347; B. G. L. Swamy et al., “Wound Healing Responses in 564/205; 514/150; 514/256; 514/297; 514/358; Monocotyledons-II. Responses to Chemical Treatments'. 514/368; 514/394; 514/397; 514/460; 514,494; Primary Examiner-Zinna Northington Davis 514/495; 514/547 Attorney, Agent, or Firm-Watson Cole Stevens Davis, 58 Field of Search ...... 546/347, 267, PL.L.C. 546/105; 514/358, 494, 495, 150, 297, 256, 332,367, 368, 394, 397, 460, 547; 57) ABSTRACT 544/242, 534/676, 773; 548/161, 164, 310.1, Traumatic acid salts, wherein B is a cation selected from: 341.5; 564/205 a) a quaternary ammonium, (56) References Cited b) a cation of a linear or branched C-C mono-, di- or trialkanolamine, U.S. PATENT DOCUMENTS c) a cation of a biologically active primary, secondary or 3,542,826 11/1970 Pacini...... 514/501 tertiary amine, 3,720,773 3/1973 Pacini...... 514/501 d) silver or zinc cation, 3,808,314 4/1974 Pacini...... 514/501 and relative pharmaceutical compositions administrable by OTHER PUBLICATIONS topical or parenteral route For the therapeutic treatment of cutaneous pathologies in which it is important to associate Chemical Abstracts, vol. 108, No. 25, Abstract No. 230587z, an a bacteriostatic, antibiotic, antifungal or an antiviral p. 600, Jun. 20, 1988. activity, to the cicatrizant effect, typical of traumatic acid. Chemical Abstracts, vol. 105, No. 17, Abstract No. 163, 456d, p. 712, Oct. 27, 1986. 10 Claims, No Drawings 5,567,716 1. 2 TRANS AND CISTRAUMATIC ACID SALTS acid seems to be due to the carboxylic group distribution HAVING CLCATRZANT ACTIVITY along the carbon chain (R. L. Goldemberg et al., Reduction ASSOCIATED TO BACTERIOSTATIC, of topical irritation, J. Chem. Soc. , 28, 667-679. 1977). ANTIVIRAL, ANTIBIOTIC OR ANTIFUNGAL Traumatic acid identification was described for the first time ACTIVITY by English and Bonnernel (The Wound Hormones of Plants, The Journal of Biological Chemistry, vol. 121, No.2, 1937), FIELD OF THE INVENTION is a vegetable hormone specifically produced by several plants as the reaction to the damage of their tissues and it is The present invention relates to traumatic acid salts for able to induce the cellular proliferation and differentiation the treatment of cutaneous pathologies where it is important O necessary to heal wound and to create the physiological to associate a bacteriostatic, antibiotic, antifungal or an stimulus for the healing of continuous solutions. antiviral activity, to the cicatrizant effect, typical of trau Traumatic acid is formed in plants by oxidation of linoleic matic acid. acid to 3-peroxylinoleic acid, which is subsequently trans formed into the Final product by enzymatic route thanks to PRIOR ART 15 the action of hydroperoxy-lyase (Zimmerman D. C., C. A. In spite of its thinness, skin plays a key role in the Condron, Identification of Traumatin, a Wound Hormone, as protection of the human organism from the external envi 12-Oxo-trans-10-dodecenoic Acid, Plant Physiol. ronment, in fact life is not possible when a large area of the 63,536-541, 1979). skin mantle is seriously damaged, as in the case of various 20 The same ability to heal wounds has been recognized in burns. the experimental animal wherein the trans-traumatic acid demonstrated (when administered by topical route) a reepi This global role of protection acts in different ways which, thelializing and antirritant action and furthermore showing individually considered, represent as many skin functions. effectiveness on an in vivo herpetic keratitis model both after Skin offers a remarkable resistance to traction, not only topic and systemic administration. thanks to horny layer cells interdigitation and to the presence of an abundant collagen filling in dermis, but also thanks to (I. Miyamoto et al., Effects of cosmetics containing bio the peculiar organization of the fibrous structure containing active substances on skin, J. Soc. Cosmet. Chem. Japan, Vol. the elastic tissue. Thanks to these characteristics, skin per 22, No.4 1989). forms a mechanical protection against traumatic insults and These pharmacological effects were subsequently recon a function of a two way barrier. In fact it allows the 30 firmed in man, where traumatic acid is proposed for burns dispersion of water (perspiratio insensibilis) and of catabo therapy since it reduces the reepithelialization times, prob lism products that are secreted by glands and conversely ably increasing the hydration layer of the cutaneous tissue, hampers the penetration of chemical substances, microor promoting cohesion between keratinocytes, reducing the ganisms of radiations present in the environment. Moreover edematous condition since it promotes the water transepi skin plays a key role in maintaining both the thermal and 35 dermic passage, and finally increasing the cells metabolism pressor homeostasis and has functions of deposit and syn in the lesion area. thesis. Finally skin has two very important characteristics Positive clinical results were also observed in psoriatic namely, the sensorial function, as it is the center of pselaph patients or in patients affected by acne, dermatitis Sebor esia and of the immunological sensitivity, since it is the rheic, neurodermatitis and itches of various origin but also immunocompetent cells center. by diseases of vital origin as for example trachoma, herpetic Cutaneous integrity maintenance allows therefore to con keratitis, verruca (Pacini 1973). With reference to the above serve unchanged the above mentioned physiological func mentioned pathologies, it is important to rely on a drug that tions. Traditional medicine has recognized over the centuries together with the cicatrizant and lenitive activities has an numerous plants used in the form of extracts or decoctions antiseptic and/or antibacterial activity and therefore broad able to aid the healing of wounds. 45 ening the traumatic acid action pattern. Still now aloe infusions and decoctions are used because of their antiphlogistic, astringent and cicatrizant effect in the treatment of burns while Asian centella and tricticum vul THE PRESENT INVENTION garis extracts are still used for the preparation of pharma 50 The applicant has unexpectedly found that trans and cis ceutical products used for the cicatrization (M. Rodriguez traumatic acid salts of formulae (I) and (II) Bigas et al., Comparative Evaluation of Aloe Vera in the Management of Burn Wounds in Guinea Pigs, Plastic and COOB Reconstructive Surgery, 1988). Booc1N-1\-1N1 S-1S In the last years a growing interest has been developed in 55 BooC1 Nu-1N1N1,N1 N cooB the attempt to isolate and to identify the substances respon sible For the pharmacological effect with the aim to under standing the difficult pharmacology of the vegetal extracts. wherein B is a biologically active cation as defined in one of Traumatic acid owing its name to its property to repair the following classes: damages in the vegetal tissue has been indicated as the active a) a quaternary ammonium cations of respectively general principle in the tricticum vulgaris extract and in the aloevera formula (III) and (IV) infusion and as such it was the object of numerous patents. (Pacini, U.S. Pat. No. 3,720,773, "Cobaltous trans-trau (III) matate for topical treatment of herpetic keratitis"). Chemi cally speaking traumatic acid or 10-dodecendioic is a linear 65 long chain dicarboxylic acid having 12 carbon atoms and an unsaturated bond: the anti-irritant activity of the traumatic 5,567,716 3 4 -continued monium, dodecyldimethyl (2-phenoxyethyl)-ammonium, R5 (IV) |(-) hexadecyl(2-hydroxychlorohexyl) dimethyl-ammonium. R6-N-R All these quaternary ammonium cations have antiseptic wherein R. R. R. and R are equal or different from and disinfectant property, the same property can also be each other and are selected from: found for the silver and zinc cations. i) a linear or branched alkyl radical having from 1 to 20 When B is the cation coming from an amine as defined in carbon atoms optionally containing in the aliphatic class (b) it is preferably selected from the group consisting chain at least one of the following groups: arylenoxy, of ethanolamine and 2-propanolamine, diethanolanime and alkylenoxy, and being optionally substituted in the di-2-propanolamine. aliphatic chain with at least one of the following 10 The biologically active amines belonging to class (c) residues: aryl, aryloxy and alkoxy groups; subclass i') are namely those having disinfectant, antiseptic ii) a cycloalkyl radical of from 3 to 10 carbon atoms; and bacteriostatic activity, and selected from the group RandR in formula (IV) form with the nitrogen atoms consisting of chlorhexidine, mafenide, hexamethylpararosa of the amine a pyridine ring, R is a C-Co linear or 15 niline, aminacrine, ethoxazene and phenazopyridine. Some branched alkyl radical, of these amines such as ethoxazene and phenazopyridine may also have analgesic and/or antiinflammatory activity. b) is a cation of a C-C linear or branched mono-, di The biologically active amines of class c) subclass ii), or trialkanolamine, having antibiotic activity are selected from the group con c) a cation of a biologically active primary, secondary or sisting of amikacin, gentamicin, kanamycin, bekanamycin, tertiary amine having a biological activity as defined in 20 neomycin, streptomycintobramycin, lincomycin, clindamy one of the following activities: cin, erythromycin, colistin, polymyxin B, tetracycline, chlo i") disinfectant, antiseptic and bacteriostatic activity, rotetracycline, rollitetracycline, Oxytetracycline, spectino ii) antibiotic activity, mycin, viomycin, bacampicyline, or stallimycin (A iii) antiviral activity, Distamycine). iv) antifungal activity, 25 d) is a cation of a metal selected From silver and zinc, not A biologically active amine of class c) subclass iii), only retain the peculiar capacity of the traumatic acid or having antiviral activity are is Tromantadine. of the considered cation, but these characteristics are The biologically active amines of class c) subclassiv) are associated and enhanced. selected from the group consisting of miconazole, econa These specific properties are to be considered of special 30 zole. chlormiconazole, chlormidazole, isoconazole, bifona interest because many antiseptic substances when topically zole, diamthazole, halethazole, hexetidine. used do not facilitate but often inhibit wound healing. The The following examples are reported for illustrative but present invention refers to their use in the preparation of not limitative purposes. pharmaceutical compositions for the therapeutic treatment EXAMPLE 1 of cutaneous pathologies wherein to the cicatrizant and 35 lenitive effect it is important to associate a bacteriostatic, Preparation of hexadecyltrimethylammonium trans-trau antibiotic, antiviral or antifungal activity. natate 2.28 g (10 mmol) of trans-traumatic acid are suspended in 50 ml of water cooled to 4 C. 7.3 g (20 mmol) of a DETALED DESCRIPTION OF THE hexadecyltrimethylamonium bromide solution in 100 ml of INVENTION water is eluted in a column cooled to 4 C. and containing The characteristics and the advantages of the traumatic 35 ml of an anionic exchange resin Dowex 1x8 generated in acid salts of the present invention are better explained in the the OH form. The eluate free from bromide anions is following detailed description. recovered in the form of traumatic acid suspension main 45 tained under stirring at 4C. The resulting solution is frozen The obtained therapeutic activities render these com and lyophilized. pounds considerably interesting in the treatment of the 7.7 g of pure product are obtained. traumatic cutaneous injuries where it is important to pro The physical chemical characteristics of hexadecyltrim mote the reepithelialization: wounds and infected wounds ethyl-ammonium trans-traumatate are the following: but also acne, dermatitis seborrheic, neurodermatitis, itches 50 of various origin, psoriasis and diseases of vital origin as for physical state: white powder example trachoma, herpetic keratitis, verrucae. In addition raw formula: Csoho2N2O4. to the therapy for these clear pathologies we have also to molecular weight: 795.38 hypothesize on the base of clinical observations relating to elemental analysis: C=75.50%; H=12.93%; N=3.52%; the "hydrating” effect of the traumatic acid, the utilization of 55 0=8.05% (calculated) these derivatives in those situations of curls reduced autosterilizing capacity by hydrolipidic film alteration and C-75.20%; H-13.11%; N=347%; O=8.22% (found) reduced resistance to the mechanical insults caused by the solubility in org. solv.:>20 mg/ml in ethanol reduced tissues elasticity. When B in the traumatic acid salts solubility in water:>10 mg/ml of general formula (I) is a quaternary ammonium cation it is 60 TLC: eluent chloroform/methanol/HO/28% NH, preferably selected from the following ones: hexadecyltri 50:40:7:3 methylamonium, dodecyltrimethylamonium and octyl-trim Rf=0.67 (traumatic acid) ethylamonium or a mixture thereof, i.e. cetrimide, ben Rf=0.18 (cetrimide). Zyldimethylhexadecylammonium, EXAMPLE 2 benzyldimethyldodecylammonium and benzyldimethyloc 65 tylammonium or a mixture thereof, namely benzethonium, Preparation of benzylmethylhexadecylammonium trans methylbenzethonium, cetylpyridinium, cetyldimethylam traumatate. 5,567,716 S 6 2.28 g (10 mmol) of trans-traumatic acid are suspended in The reaction yield is 2.65 g of dry product. 50 ml of water cooled to 4 C. The physical chemical properties of Zinc trans-trau 8.28 g (20 mmol) of a benzylmethylhexadecylammonium mateate are the following: chloride solution in 90 ml of water and 10 ml ethanol are physical state: white amorphous powder eluted in a column cooled to 4°C. and containing 35 ml of an anionic exchange resin DoweX 1x8 generated in the raw formula: CHOZn OH form. The eluate free from chloride anions is recovered molecular weight: 291,67 in the form of a traumatic acid suspension maintained under elemental analysis: C-49.42%; H=6.22%; O=21.94%; stirring at 4 C. The resulting solution is frozen and Zn=22.42% (calculated): C=49.11%; H=6.23%; lyophilized. 10 O=21.82%; Zn=22.84%, (found) 9.2 g of pure product are obtained. traumatic acid: 78.27% (as free acid) The physical chemical characteristics of the benzylmeth ylhexadecylammonium trans-traumatate thus obtained are water solubility: poorly soluble (>10 mg/ml in 5% NH) the following: organic solvent solubility: poorly soluble in DMSO and ethanol physical state: amorphous white deliquescent powder 15 TLC: eluent chloroform/methanol/water/28% NH, raw formula: C2HoN2O4. 50:40:7:3; Rf=0.67 (traumatic acid) molecular weight: 947.58 elemental analysis: C=78.59%; H=11.70%; N=2.96%; EXAMPLE 5 O=6.75% (calculated): C=78.30%; H=11.81%; 20 N=2.89%; O=6.81% (found) Preparation of benzethonium trans-traumatate 2.28 g of trans-traumatic acid (10 mmol) are suspended in 50 ml of solubility in org. solv.: >20 mg/ml in ethanol water cooled to 4°C. solubility in water: >10 mg/ml A solution of 9,0g of Benzethonium chloride (20 mmol) TLC: eluent:chloroform/methanol/HO/28% NH, in 90 ml of water is eluted through a column cooled to 4°C. 50:40:7:3 Rf-0.67 (traumatic acid) RF=0.6.3 (ben 25 and containing 35 ml of OH- DoweX 1.x8 resin. Zyldimethyl-hexadecyl-ammonium). The eluate free from chlorides is then frozen and lyo EXAMPLE 3 philized. The reaction yield is 10.5g of dry product. The physical chemical properties of benzethonium trans Preparation of silver trans-traumatate traumatate are the following: 2.28 g of trans-traumatic acid (10 mmol) are suspended in 30 physical state: deliquescent solid 20 ml water at 4°C., neutralized using NaOH 1N, sheltered from light. The resulting suspension, kept at 4 C., is raw formula: C6Ho2N2Os exposed to a nitrogen stream and a solution of 3.4 g AgNO molecular weight: 1051.56 in 20 ml water is slowly added drop by drop under continu elemental analysis: C=75.39%; H=9.78%; N=2.66%; ous stirring. 35 O=12.17%; (calculated): The resulting precipitate is separated by filtration, washed C-75.31%; H-9.84%; N=2.58%; O=12.27%; (found) three times using 10 ml of cool water and finally dried under traumatic acid: 21.71% high vacuum. The reaction yield is 4.1 g of dry product. The physical chemical properties of silver trans-trau benzethonium: 85.23% (as benzethonium chloride) matate are the following: 40 water solubility: >10 mg/ml physical state: white amorphous powder organic solvent solubility: >10 mg/ml in ethanol raw formula: CHOAg2. TLC: eluent chloroform/methanol/water/28% NH 50:40:7:3; Rf-0.67 (traumatic acid): eluent ethanol/ molecular weight: 442,02 waterfacetic acid 70:20:10; Rf=0.53 (benzethonium) elemental analysis: C=32.61%; H-4.10%; O=14.48%; 45 Ag=48.81%. (calculated): =32.29; H=4.21%; O=14.82%; Ag=48.68%. (found) EXAMPLE 6 traumatic acid 51.65% (as free acid) Preparation of tobramycin trans-traumatate water solubility: poorly soluble (>10 mg/ml in 5% NH3) 11.4 g of trans-traumatic traumatic acid (50 mmol) are 50 suspended in 200 ml of cool water at 4 C., A solution of organic solvent solubility: poorly soluble in DMSO and 14.3 g of tobramycin sulfate (20mmol) in 200 ml of water ethanol is eluted through a column cooled to 4°C. containing 150 ml TLC: eluent chloroform/methanol/water/28% NH of OH- Dowex 1x8 resin. The eluate free from sulfate is 50:40:7:3; Rf-0.67 (traumatic acid) collected into the suspension of traumatic acid kept under EXAMPLE 4 55 continuous stirring at 4°C. The resulting solution is frozen and lyophilized. Preparation of zinc trans-traumatate The reaction yield is 20.2 g of dry product, 2.28 g of trans-traumatic acid (10 mmol) are suspended in The physical chemical properties of tobramycin trans 20 ml cool water at 4°C. and neutralized using NaOH lN. traumatate are the following: A solution of 2.88 g ZnSO, eptahydrate (10 mmol) in 20 physical state: white amorphous powder ml water is slowly added drop by drop to the resulting solution, kept at 4 C. and under continuous stirring. raw formula: Coghla NoOas The resulting mixture is heated to 40° C. for 3 hours and molecular weight: 2076.5 then cooled to 4° C. for further 15 hours. elemental analysis: C-55.53%; H=8.45%; N=6.75%; The precipitate is recovered by filtration, washed three 65 O=29.28%; (calculated): C=55.26%; H=8.68%; times using 10 ml of cool water and finally dried under high N=6.96%; O=29.10%; (found) WaCl. traumatic acid: 54.97% (as free acid) 5,567,716 7 8 tobramycin: 68.64% (as tobramycin sulfate) EXAMPLE 9 water solubility: >10 mg/ml Preparation of Erythromycin trans-traumatate organic solvent solubility: >10 mg/ml in DMSO 2.28g of trans-traumatic acid (10 mmol) are suspended in TLC: eluent chloroform/methanol/water/28% NH 100 ml of waterlethanol 2:1 mixture at 4° C. 14.7 g of 50:40:7:3; Rf=0.67 (traumatic acid): eluent chloro 5 erythromycin free base (20 mmol) are added and the result form/methanol/28% NH 50:40:10; Rf=0.05 (tobramy ing mixture is kept under stirring at 4 C. for 3 hours and cin) then heated to 25° C. overnight. 200 ml water ape then EXAMPLE 7 added. The mixture is concentrated under vacuum to a volume of about 100 ml and cooled to 4°C. Preparation of gentamycin trans-traumatate 10 The resulting precipitate is recovered by filtration, washed 11.4 g of trans-traumatic acid (50 mmol) are suspended in three times using 10 ml of cool water and finally dried under 200 ml of water cooled to 4 C. high vacuum. The reaction yield is 16.3 g of dry product. A solution of 14.4 g gentamycin sulfate (20 mmol) in 200 The physical chemical properties of erythromycin trans ml of water are eluted through a column cooled at 4°C. and traumatate are the following: containing 150 ml (OH- Dowex 1x8 resin. The eluate free 15 from sulfate is collected in the form of a traumatic acid physical state: white amorphous powder suspension kept under continuous stirring at 4 C. The raw formula: C8H19N2Oao resulting solution is frozen and lyophilized. The reaction molecular weight: 1696.19 yield is 20.6 g of dry product. elemental analysis: C=60.90%; H=9.15%; N=1.65%; The physical chemical properties of gentamycin trans 20 O=28.3%; (calculated): C=60.60%; H-9.32%; traumatate are the following: N=1.58%; O-28.5%; (found) physical state: white amorphous powder traumatic acid: 13.46% (as free acid) raw formula: Co2H8NoC)34 erythromycin; 86.54% (as free base) molecular weight: 2096.7 25 water solubility: poorly soluble elemental analysis: C=58.43%; H=8.94%; N=6.68%; organic solvent solubility: >10 mg/ml in DMSO and O-25.95%; (calculated): C=58.21%; H-9.05%; ethanol N=6.59%; O-26.17%; (found) TLC: eluent chloroform/methanol/water/28% NH traumatic acid: 54.44% (as free acid) 50:40:7:3; RF=0.67 (traumatic acid): eluent chloro gentamycin: 68.975 (as gentamycin C1 sulfate) 30 form/methanol/water/28% NH, 80:25:2:1; Rf=0.86 water solubility: >10 mg/ml (erythromycin) organic solvent solubility: >10 mg/ml in DMSO TLC: eluent chloroform/methanol/water/28% NH EXAMPLE 10 50:40:7:3; RF=0.67 (traumatic acid): eluent chloro 35 Preparation of rollitetracycline trans-traumatate form/methanol/28% NH 1:1:1 ninhydrin as indicator: 2.28g of trans-traumatic acid (10 mmol) are suspended in three stains are observed like in gentamycin sulfate 200 ml of water at 4°C. under nitrogen stream and sheltered used as standard sample. From light. A solution of 10.6 g rollitetracycline (20 mmol) EXAMPLE 8 in 100 ml H2O is slowly added drop by drop in 30 minutes 40 to the above suspension Preparation of lincomycin trans-traumatate The resulting mixture is frozen and lyophilized. 2.28 g of trans-traumatic acid (10 mmol) are suspended in The reaction yield is 12.6 g of dry product. 100 ml of water cooled to 4° C. A solution of 8.86 g of The physical chemical properties of rollitetracycline tran lincomycin hydrochloride (20 mmol) in 100 ml water is straumarate are the following: eluted through a column cooled to 4°C. containing 35 ml of 45 physical state: amorphous yellow-brown powder OH- Dowex 1x8 resin. The eluate free from chlorides is collected in the form of a suspension of traumatic acid kept raw formula: C5H8N6O20 under continuous stirring at 4 C. The resulting solution is molecular weight: 1283.5 frozen and lyophilized. The reaction yield is 10.1 g of dry elemental analysis: C=61.77%; H=6.75%; =6.55%; product. 50 O=24.93%; (calculated): C=61.62%; H=6.90%; The physical chemical properties of lincomycin trans N=6.49%; O=24.99%; (found) traumatate are the following: traumatic acid 17.79% (as free acid) physical state: white amorphous powder rolitetracycline 82.21% (as free base) raw formula: CashssNOS 55 water solubility >10 mg/ml molecular weight: 1041,38 organic solvent solubility: >10 mg/ml in DMSO elemental analysis: C=55.36%: H=8.52%; N=5.38%: TLC: eluent chloroform/methanol/water/28% NH O=24.58%: S=6.16% (calculated): C=54.98%; 50:40:7:3; Rf=0.67 (traumatic acid): eluent butanol/ H-8.69%; N=5.31%; O=25.015 S=6.01 (found) acetic acid/water 4:2:2; Rf-0.53 (rolitetracycline) traumatic acid: 21.92% (as free acid) 60 lincomycin: 85.08% (as lincomycin hydrochloride) EXAMPLE 11 water solubility: >10 mg/ml Preparation of chlorhexidine trans-traumatate organic solvent solubility: >10 mg/ml in DMSO 2,28g of trans-traumatic acid (10 mmol) are suspended in TLC: eluent chloroform/methanol/water/28% NH 65 100 ml of a mixture waterlethanol 2: 1 at 4° C., 5,05 g of 50:40:7:3; Rf=0.67 (traumatic acid): eluent ethanol/ chlorhexidine base (10 mmol) are added and the mixture is waterfacetic acid 70:20:10 Rf=0.61 (lincomycin) kept under stirring at 4° C. for hours and then at 25° C. 5,567,716 9 10 overnight. 200 ml of water are added and the mixture is The physical chemical properties of miconazole trans concentrated under vacuum at the volume of about 100 ml traumatate and Finally again cooled to 4°C. The resulting precipitate is physical state: white amorphous powder recovered by filtration, washed three times using 10 ml of cool water and finally dried under high vacuum. 5 raw formula: Cashin,00ls The reaction yield is 6.8g of dry product. molecular weight: 1060.6 The physical chemical properties of chlorhexidine trans elemental analysis: C-54.36%; H-4.56%; N=5.28%; traumatate are the following: O=9.05%; Cl=26.74 % (calculated): C=54 52%; physical state: white amorphous powder H-4.61%, 61%; N=5.17%; C-26.50% (found) raw formula: CHSoNoOCl, 10 traumatic acid: 78.47% (as free acid) molecular weight: 733.75 ethanolamine: 21.53% (as free base) elemental analysis: C=55.66%; H=6.87; N=19.09%; water solubility: >10 mg/ml O=8.72%; Cl=9.66% (calculated): C=55.38%; organic solvent solubility: >10 mg/ml in ethanol H=6.99%; N=18.92%; O=8.96% C-9.75% (found) 15 TLC: eluent chloroform/methanol/water/28% NH, 50: traumatic acid: 31.11% (as free acid) 40: 7:3; RF=0. 67 (traumatic acid): eluent chloroform/ chlorhexidine: 68.98% (as free base) methanol/water/28% NH 80:25:2:1; Rf=0.93 (micona water solubility: poorly soluble zole)

organic solvent solubility: >10 mg/ml in DMSO 20 TLC: eluent chloroform/methanol/water/28% NH BIOLOGICAL ACTIVITY 50:40:7:3; Rf-0.67 (traumatic acid): eluent chloro For purposes of clarity the product tested in the following form/methanol/28% NH-50:40:10Rf=0.77 (chlorhexi pharmacological trials are those prepared as described in the dine) above reported chemical preparations examples. Therefore 25 they are all salts of trans-traumatic acid. These trials have the aim to evaluate both the cicatrizant EXAMPLE 12 activity of these compounds with respect to that of traumatic Preparation of ethanolamine trans-traumatate acid and the antibacterial activity with respect to a compari 2.28g of trans-traumatic acid (10 mmol) are suspended in son molecule having the same activity in suitable trials 100 ml of water at 4 C. A solution of 1.23g ethanolamine 30 carried out in vitro. as free base in 20 ml water, under continuous stirring is 1. Effect of cellular proliferation evaluated on fibroblasts of slowly added drop by drop in 30 minutes to the resulting the traumatic acid salts. Suspension. Experiments on 3T3 fibroblasts The resulting mixture is frozen and lyophilized. 1.la Materials and methods. The reaction yield is 3.5g of dry product. 35 Cells preparation Mice 3T3 fibroblasts cultures sown at a 40.000/ml con The physical chemical properties of ethanolamine trans centration on plates having 6 wells in a DMEM culture traumatate are the following: medium enriched with 10% calf serum are used. After 24 physical state: white amorphous powder hours culture, the 90 prepared wells are divided in order to raw formula: CH3N2C allow the analyses of the compounds of the invention, every molecular weight: 350.46 compound to be tested being used respectively at a concen tration of 0.1, 1 and 10 ug/ml by subsequent evaluations elemental analysis: C=54.84%; H=9.78%; N-7.99%; after 48 and 72 hours of incubation. O=27.39%; (calculated): C-54.58%; H=9.84%; Compounds solubilization: N=8.02%; O=27.62%; (found) Hexadecyltrimethylammoniun (ETA) traumatare: solubi traumatic acid: 65.14% (as free acid) 45 lized in water and brought to the desired final concen ethanolamine: 34.86% (as free base) trations of 0.1. 1 and 10 g/ml by progressive dilutions; water solubility: >10mg/ml Ethanolamine traumatate: solubilized in water and organic solvent solubility: >10 mg/ml in ethanol brought to the final concentrations of 0.1, 1 and 10 TLC: eluent chloroform/methanol/water 28% NH 50 ug/ml by progressive dilutions; 50:40:7:3; Rf=0.67 (traumatic acid) Traumatic acid: neutralized with sodium hydroxyde and solubilized in water, then brought to the final concen trations of 0.1, 1 and 10 ug/ml by progressive dilutions EXAMPLE 13 with the culture medium. Preparation of miconazole trans-traumatate 55 The compounds are added to the cells after 24 hours 9.58g of miconazole nitrate (20 mmol) are suspended in culture. The cells number is measured by colorimetry with the neutral red intravital stain after 24, 48, 72 hours of 300 ml of water and 5 ml of 30% NHOH are added. The incubation. resulting mixture is extracted 3 times using each time 100 ml Results: of chloroform and the organic phases ape washed 3 times These compounds act according to a dose-effect rela with 50 ml of water, submitted to anhydrous condition using tion. Particularly hexadecyltrimethylammonium (ETA) trau Na2SO and then collected together and evaporated under matare, ethanolamine traumatare, and traumatic acid at 0.1 WaClil. ug/ml concentration are able to affect cells proliferation 2.28 g of trans-traumatic acid (10 mmol) are added to the according to a not statistically significant trend. At the raw residue solubilized in 200 ml of ethanol. The resulting concentration of 1 ug/ml hexadecyltrimethylammonium mixture is evaporated under vacuum and the residue dried 65 (ETA) traumarate>ethanolamine traumatare=traumatic acid under high vacuum, are able to increase cellular proliferation. At the highest The reaction yield is 20.5g of dry product. concentration (10 ug/ml) the compounds tend to reduce or at 5,567,716 11 12 least not to increase cellular proliferation. From a biological inoculated, preparing a final solution of 10 cfu/g (colony standpoint the compounds give more significant results after forming unities) of the following strains: 48 hour (table 1.1-a), although already after 24 hours a higher effect is observed if compared to that obtained with control cultures. A side control experiment is also carried out Pseudomonas Aeruginosa ATCC 35422 in which the cultures confluence is evaluated: the cultures Cepacia ATCC 25416 Maltophilia ATCC 13637 treated with hexadecyltrimethylammonium traumatare and Staphylococcus Aureus ATCC 65380 ethanolamine traumatarestraumatic acid reduce the time Staphylococcus Epidermidis ATCC 14990 normally necessary for the cells to reach the confluence. Streptococcus Fecalis ATCC 29212 These data demonstrate that these compounds are able to Escherichia Coli ATCC 35248 stimulate cicatrization processes. 10 Candida Albicans ATCC 10231 1.1b Materials and methods Aspergillus Niger ATCC 16404 Cell growth Mouse Balb/C 3T3 fibroblasts were grown in DMEM Plates preparation medium supplemented with penicillin (100 units/ml) and streptomycin (0.1 mg/ml). For experimental purposes, cells Every plate contains 20 ml of a medium where 2 equi are routinely inoculated in 6.4 um diameter culture dishes 15 distant wells having a standard diameter are Formed and in (5000cells/dish) and cultured overnight. After 24 hours the each of these wells 200 ug of the above described bacterial culture medium was decantated and the compounds to be solution are inoculated. 2 plates are prepared for every tested are added in the presence of 10% foetal calf serum couple of compounds to be tested at different concentrations, (FCS). and For every strain, and each of these plates is compared The cells are incubated at 37° C. in 5% CO, air for 4 (to), 20 with a white (sterile demineralized water inoculum). After 24 (t1) and 48 (t2) hours before cell density measurement by the inoculum, the plates are placed in the refrigerator to colorimetry using crystal violet (exposure for 15 minutes, favour diffusion and temporaneously blocking bacterial room temperature, absorbance 570mm). growth. Compounds solubilization The compounds hexadecyltrimethylammonium trau The compounds to be tested are solubilized in DMEM+ 10%FCS and DMSO to obtain the final concentration of 25 matate and benzyldimethylhexadecylammonium traumatare 10M, 10 Me 107M in 0.05% DMSO. respectively versus hexadecyl trimethylammonium bromide Results and benzyldimethylhexadecylammonium chloride are solu The cells number is measured 4, 24 and 48 hours after bilized in demineralized water to Form solutions at the seeding. The cell growth is evaluated as t2/t1 ratio (%). The concentrations: 0.1; 0.5; and 1%. results described in table 1.1-b show that lincomycin 30 Parameters traumatate>rolitetracycline traumatate?erythromycin trau The antibacterial effect is evaluated by measuring bacte matate are capable to promote cell growth at all the tested rial diffusion halo around the well vs white (halo absence). doses and this effect appears more consistentif compared to Results the corresponding one of the parent compound traumatic acid. The analysis of the results evidences that the two com 35 pounds of the invention hexadecyltrimethylammonium trau 1.2 Mice fibroblast L. 929 cell culture matare and benzyldimethylhexadecylammonium traumatare Materials and methods have an antibacterial and bacteriostatic activity profile com Cell growth parable to that of the comparison products hexadecyltrim Mice fibroblast cells L929, clone derived from connective tissue, are suspended in Eagle's MEM with penicillin ethylammonium bromide and benzyldimethylhexadecylam (100U/ml)-streptomycin (0.1 mg/ml), seeded in culture 40 monium chloride, indicating that the salification has not dishes (10000cells/dish) and cultured overnight. After 24 altered the original biological properties (Table 2. a). hours the culture medium was decantated and the tested 2.b EVALUATION OF BACTERIOSTATIC MINIMAL compounds are added. The cellular density is measured after CONCENTRATION (BMC) 4 (tO) and 24 (t1) hours of incubation at 37° C. in 5% CO, Materials air by colorimetry using crystal violet (exposure for 15 45 1. Culture Medium: Tryprone Soy Broth (TBS) - Bioge minutes, room temperature, absorbance 570 nm). netics Compounds solubilization 2. Saline solution: 0.9 g NaCl in 100 ml demineralized The compounds Benzethonium traumarate, gentamycin and sterile water traumatare acid are solubilized in water at the concentration of 10M, 107M and 10M, while traumatic acid was 50 3. McFarland solution: 1 ml BaCl. 1.175% in 99.5 ml 1% solubilized in DMSO at the above mentioned concentra HSO tions. Methods Results Bacterial Suspension preparation: The cells number is measured at 4 (tO) and 24 (t1) hours Microbic standard strains from American Type Culture from the seeding. The cell growth is evaluated at t1/t0 rate Collection (ATCC) were used, chosen among aerobic Gram (%). The results described in table 1.2 show that benzetho 55 positive and negative as follows: nium traumatare) gentamycin traumarate ape capable to promote cell growth (proliferation) according to dose-effect Pseudomonas Aeruginosa ATCC 5422 relationship and this effect is higher than traumatic acid Safilococcus Aureus ATCC 5380 effect. The above experimental data indicate that the salifi Stafilococcus Epidermis ATCC 14990 cation process can also increase the proliferative effect in 60 Bacilius Subtilis ATCC 6633 comparison with parent compound. Escherichia Coli ATCC 35218 2. Antibacterial activity of the traumatatic acid salts 2.a Candida Albicans ATCC 1023 diffusion method in agar-germs Materials and methods The broths culture of the different species are prepared Bacterial suspention preparation: 65 growing the standard strain (kept in Tryptic Soy Agar (TSA) In a Tryprone Soy Agar diffusion medium ATCC (Ameri slent at 5° C) in Tryptone Soy Broth (TSB) for 12 hours at can Type Culture Collection) standard bacterial strains are 37° C. 10 ml are then taken up from each culture, centrifu 5,567,716 13 14 gated for 3 times at 3500 PM washing each time the Evaluation of Bacteriostatic Minimal Concentration (BMC): Sediment using 10 ml of sterile saline solution. The sediment 50 ul of bacterial suspension are added to the tube is resuspended in the saline solution until obtainment of a containing the compounds to be tested and incubated at 37 turbidity corresponding to the McFarland standard solution (the suspension title must be 107-10 UFC/ml). C.: the densitometric determination is made in comparison Preparation of the solutions of the compounds to be tested: with McFarland standard solution 24 and 48 hours later. The stock solutions (5% in sterile water, added with 1N After the incubation time the suspensions ape gently HCl in case the salts are poorly soluble in water) of the agitated; 50 ul of each suspension are inoculated in tubes traumatic acid salts are prepared and kept at 5° C. containing 5 ul of TSB and incubated at 37° C.; the Hexadecyltrimethyl ammonium traumatare O densitometric determination is made 48 hours later. Benzyldimethylhexadecyl ammonium traumatare BMC is defined by comparison between the two densi Zinc traumarate tometric evaluation. Silver traumarate Results Chlorhexidine traumarate All the examined compounds show bacteriostatic effect as Benzethonium traumatate 15 Gentamicin traumatare described in Tables 2.b)1-6. This effect is specific for the Erythromycin traumatare different bacterial strains tested. The bacteriostatic effect of Tobramycin traumatare each salt is similar to stechiometric concentration of the Lincomycin traumatare respective negative ion. Rolitetracycline traumarate 20 The above experiment suggest that the new salts of traumatic acid here described, capable to promote cell The compounds, dissolved as stock solution, were added proliferation as shown before, possess also bacteriostatic to test tubes containing 5 ml TBS in order to obtain the effect. Furthermore the above experiment show that the desired Final concentration 10000; 5000; 1000; 200; 40; 2; salification process does not alter the specific effect that the 0.4; 0.05 g/ml (culture medium maximal dilution=1/10). anion contribute.

TABLE 1 a Effects of hexadecyltrimethylammonium (ETA) traumatate and ethanolamine traumatate with respect to traumatic acid on the cellular proliferation of fibroblasts. The cells numeration is accomplished by colorimetric method after 24, 48 and 72 hours of incubation. Concentrations (pg/ml) Substance 0. 10 0.0

24 hours of incubation (Control 0.579 + 0.021) ETA 0.575 0.041 0.619-0.033 0.491 - 0.02 tallatate ethanolamine 0.562 - 0.013 0.609 - 0.033 0.553 - 0.027 taatate traumatic 0.486 - 0.029 0.5100.008 0.543 - 0.019 acid 48 hours of incubation (Control 1.334 + 0.050) ETA 1553 - 0.061 1.50 0.081 0.842 + 0.078(**) raumatate ethanolamine 145 - 0.06 1435 0.082 415 - 0.132 tallatate traumatic acid 1497 - 0.087 1431 0.047 14420,060. 72 hours of incubation (Control 2.360+ 0.125) ETA 2.501 + 0.131 2.503 - 0.100 1.385 + 1.63(**) tallatate ethanolamine 2.382 - 0.039 2.463 0.00 2.408: 0.085 traumatate traumatic 2.492 - 0.070. 2.349 it 0.25 2.446 - 0.128 acid Significant increase of neutral red incorporation

Significant reduction of neutral red incorporation 5,567,716 15 16 TABLE 1.1b TABLE 1.1b-continued Effect of lincomycin traumatate, erythromycin traumatate and Effect of lincomycin traumatate, erythromycin traumatate and rolitetracycline traumatate compared to traumatic acid on cell rolitetracycline traumatate compared to traumatic acid on cell growth in mice 3T3 ribroblast colture 5 growth in mice 3T3 ribroblast colture Cell density at different Cell density at different time (hours) cell growth time (hours) cell growth 4(tO) 24(t1) 48(t2) t2ft 4(t0) 24(t1) 48(2) t2ft1 10 Control 0.300 0.481 0.849 76.776 --052 +-102 --.108 --44.70 --.045 --075 --.139 +-15.348 Traumatic acid 10-5M. 0.478 0.317 0.657 112.273 TABLE 1.2 --045 --,074 --.113 --17.727 10-6M 0.495 0.320 0.657 95.638 15 Cell growth evaluation in mice ribroblast L929 colture: effect of --.096 --.055 ---.113 --24.834 Benzethonium Traumatate and gentamycin traumatate in 10-7M 0.459 0.323 0.740 135.554 comparison with traumatic acid, added at different doses. --.057 +-07 --,097 +-33,092 Lincomycin traumatate cell density cell growth 10-5M 0.361 0.228 0.693 195.78 20 4h(0) 24h(t)) t110 --049 --.045 --065 +-20.35 10-6M 0.352 0.210 0.734 246.29 Control 0.335-h-024 0.407 -- 040 22.4 -- 16.29 --.062 --,041 --.127 --4.87 Traumatic acid 10-7M 0.384 0.209 0.705 247.68 --,023 --.069 --.22 -58.22 10-6M 0.272 -- 028 0.404 -- 008 50.5 - 15.18% is Erythromycin traumatate 25 10-7M 0.261 -- 017 0.401 -i-.039 53.63 - 12.67* * ama-am- 10-8M 0.327 -.022 0.345 – 043 6.95 -- 19.21 10-5M 0.370 0.365 0.919 159.16 Benzethonium traumatate --.036 --,025 +-107 --10.42 10-6M 0.352 0.369 0.921 148.64 10-6M 0.209-017 0.402 - 024 91.06 - 23.74** --047 --,027 --07 --4.66 10-7M 0.266 -- 009 0.407 -- 026 52.16 -- 1.638 10-7M 0.344 0.368 1.074 196.81 30 0-8M 0.268 -i-.014 0.363 - 027 35.6+ - 7.57 --,055 --013 --085 --10.28 Gentamycin traumatate Rolitetracycline traumatate -----m- 10-6M 0.251 +-027 0.405 -- 016 63.68 - 17* * 10-5M 0.46 0.255 0.678 166.39 10-7M 0.264 - 003 0.357 - 019 35.3 - 6.24 --.074 --,068 --.243 --22.91 0-8M 0.299-009 0.385-06 29 - 5.03 10-6M 0.475 0.356 0.660 186.69 35 --.031 --059 --170 --5.13 *p < 0.05; 10-7M 0.482 0.287 0.828 185.44 **p < 0.01

TABLE 2.a. Antibacterial activities of the salts hexadecyltrimethylammonium (ETA) traumatate and benzyldimethylhexadecylammonium (BMA) traumatate respectively versus ETA bromide and BMA chloride. A Concentrations sol. 1% sol. 0.5% sol. 0.1% Tested ETA ETA ETA ETA ETA ETA solutions Traumatate Bromide Traumatate Bromide Traumatate Bromide Standard strains

Pseudomonas n.h. n.h. n.h. n.h. n.h. n.h. Aeruginosa ATCC35422 PS 22 22 19 19 18 18 Cepacia ATCC.25416 PS 21 21 5 15 n.h. n.h. Maltophila ATCC.13637 Stafilococcus 24 24 24 24 21 21 Aureus ATCC.65380 Stafilococcus 22 22 22 22 20 20 Epidermidis ATCC.14990 Streptococcus 22 22 22 22 20 20 Fecalis ATCC.29212 E. Coli 15 15 13 13 n.h. n.h. ATCC,35218 5,567,716 17 18 TABLE 2a-continued Antibacterial activities of the salts hexadecyltrimethylammonium (ETA) traumatate and benzyldimethylhexadecylammonium (BMA) traumatate respectively versus ETA bromide and BMA chloride. Candida 25 25 24 24 19 9 Albicans ATCC.O231 Aspergillus 22 22 22 22 19 19 Niger ATCC 16404 B

Concentrations sol. 1% Sol. 0.5% sol. 0.1%

Tested BMA BMA BMA BMA BMA BMA solutions Traumatate Chloride Traumatate Chloride Traumatate Chloride Standard strains

Pseudomonas 14 14 2 12 n.h. n.h. Aeruginosa ATCC35422 PS 15 15 15 15 2 12 Cepacia ATCC 25416 PS 15 15 n.h. n.h. n.h. n.h. Maltophila ATCC-13637 Stafilococcus 22 22 22 22 17 17 Aureus ATCC.65380 Stafilococcus 19 19 17 18 15 15 Epidermidis ATCC.14990 Streptococcus 15 15 14 14 2 2 Fecalis ATCC.2922 E. Coli 13 13 n.h. n.h. n.h. n.h. ATCC.3528 Candida 16 6 15 5 3 13 Albicans ATCC.1023 Aspergillus 19 19 16 16 12 13 Niger ATCC.6404 n.h.. it no halo

TABLE 2.b)1 Psaeruginosa ATCC 35422 MNIMAL BACTERIAL COMPOUND CONCENTRATION Hexadecyltrimethyl Ammonium Traumatate >0,000 Benzyldimethylhexadecyl Ammonium Traumatate 10,000.00 Zinc Traumatate >10,000 Silver Traumatate 40.00 Chlorhexidine Traumatate 40.00 Benzethonium Traumatate 200.00 Gentamycin Traumatate 1,000.00 Erythromycin Traumatate 10,000.00 Tobramycin Traumatate 5,000 Lincomycin Traumatate 10,000.00 Rolitecracycline Traumatate 10,000.00 Control: positive

60 5,567,716 19 20 TABLE 2.b)2 Staureus ATCC 65380 MINIMAL BACTERIAL COMPOUND CONCENTRATION Hexadecyltrimethyl Ammonium Traumatate 1,000.00 Benzyldimethylhexadecyl Ammonium Traumatate 1,000.00 Zinc Traumatate >10,000 Silver Traumatate 40.00 Chlorhexidine Traumatate 8.00 Benzethonium Traumatate 2.00 Gentamycin Traumatate 1,000.00 Erythromycin Traumatate 10,000.00 Tobramycin Traumatate 1,000.00 Lincomycin Traumatate 10,000.00 Rolitetracycline Traumatate 5,000.00 Control: positive

TABLE 2.b)3 Ecoli ATCC 35218 MINIMAL BACTERIAL COMPOUND CONCENTRATION Hexadecyltrimethyl Ammonium Traumatate 200.00 Benzyidimethylhexadecyl Ammonium Traumatate 200.00 Zinc Traumatate >10,000 Silver Traumatate 40.00 Chlorhexidine Traumatate 8.00 Benzethonium Traumatate 40.00 Gentamycin Traumatate 200.00 Erythromycin Traumatate 10,000.00 Tobramycin Traumatate 200.00 Lincomycin Traumatate 10,000.00 Rolitetracycline Traumatate 1,000.00 Control: positive

TABLE 2.b)4 Stepidermidis ATCC 14990 MNIMAL BACTERIAL COMPOUND CONCENTRATION Hexadecyltrimethyl Ammonium Traumatate 8.00 Benzyldimethylhexadecyl Ammonium Traumatate 2.00 Zinc Traumatate >10,000 Silver Traumatate 200.00 Chlorhexidine Traumatate 2.00 Benzethonium Traumatate 2.00 Gentamycin Traumatate 200.00 Erythromycin Traumatate 10,000.00 Tobramycin Traumatate n.t. Lincomycin Traumatate 200.00 Rolitetracycline Traumatate 200.00 Control: positive 5,567,716 21 22 TABLE 2.b)5 B. subtilis ATCC 6633

MINIMAL BACTERIAL COMPOUND CONCENTRATION Hexadecyltrimethyl Ammonium Traumatate 2.00 Benzydimethylhexadecyl Ammonium Traumatate 1,000.00 Zinc Traumatate >10,000 Silver Traumatate 200.00 Chlorhexidine Traumatate 2.00 Benzethonium Traumatate 8.00 Gentamycin Traumatate 40.00 Erythromycin Traumatate 1,000.00 Tobramycin Traumatate El..t. Lincomycin Traumatate 1,000.00 Rolitetracycline Traumatate 40.00 Control: positive

TABLE 2.b)6 Calbicans ATCC 1023 MINMAL BACTERIAL COMPOUND CONCENTRATION Hexadecyltrimethyl Ammonium Traumatate 2.00 Benzyidimethylhexadecyl Ammonium Traumatate 2.00 Zinc Traumatate >10,000 Silver Traumatate >10,000 Chlorhexidine Traumatate 2.00 Benzethonium Traumatate 2.00 Gentamycin Traumatate >10,000 Erythromycin Traumatate >10,000 Tobramycin Traumatate ...t. Lincomycin Traumatate 10,000.00 Rolitetracycline Traumatate 1,000.00 Control: positive

Conclusions For these pathologies both the parenteral (vials for intra The reported results clearly evidence that the compounds 40 muscular use) and the topical administration (creams, oint of the present invention by salification of traumatic acid are ments, gels and solutions) are foreseen. able to determine a specific and considerable cicatrizant The necessary dose to perform the therapeutic effect activity associated with a marked antibacterial activity. varies depending on the type and seriousness of the damage These effects may be advantageously used in the patholo and on the considered patient (age, concomitant patholo 45 gies). A therapeutic dosage is preferable being comprised gies wherein it is desirable to associate a stimulation of between 100 and 300 mg/die with periods variable in tissue reparative processes with an antibacterial effect, also relation to the type of pathology and anyway not shorter than considering that many topical antiseptic agents used in two weeks. surgical practice do not facilitate, but often inhibit injuries Reported hereinbelow for illustrative but not limitative healing. (Meyers, Jawetz and Goldfien Farmacologia Medica purpose are the following examples of therapeutic compo chap. 58 Ed. Piccin, 1975). sitions containing as active principle some traumatic acid The salts of the present inventions are therefore useful in salts of formula (I) human therapy fop the treatment of injuries and infected injuries as a consequence of a surgical operation, fistulae, 55 necrotic processes, ulcerodystrophic alterations ( torpid EXAMPLE 1. sores, bedsores, burns, fistulous stabs and rhagades) or, Cream anyway, situations requiring the reaction of the process of Every tube contains 100 g cream epithelial neoformation such as acne, dermatitis seborrheic, neurodermatitis, itches also of allergic nature or dermic 60 ETA traumatate 1.75 g phenomena of intoxication without excluding affections of Cetostearyl alcohol 7.00 g vital origin such as trachoma, herpetic keratitis and verrucae. Isopropylmyristate 7.00 g Moreover pathologies are to be added being more simply Liquid paraffin 7.00 g related to a delayed or altered dermic layer renewal or White beeswax 3.00 g Glycerol 3.00 g involving alterations of the cutaneous hydrolipidic layer 65 Cetomacrogol 100 2.20 g (e.g. cutaneous ageing, damage by make-up or cosmetics Perfume test. 34152 (ICSA) 0.10 g excess). 5,567,716 23 24 -continued aliphatic chain with at least one of the following Butylhydroxytoluene 0.01 g residues: aryl, arlyoxy and alkoxy groups; and Purified water q.s. to 100 g ii) a cycloalkyl radical of from 3 to 10 carbon atoms; Rs and R in formula (IV) form with the nitrogen atom 5 a pyridine ring, R is a C-Co linear or branched alkyl radical; EXAMPLE 2 b) a cation of linear or branched C-C mono-, di-, or Nebulizer solution tri-alkanolamine; c) a cation of a primary, secondary or tertiary amine Every bottle contains 10 ml solution: 10 belonging to one of the following subclasses: i") amines with disinfectant, antiseptic and bacterio ETA traumatate 175 mg static activity, selected from the group consisting of NaCl 90 mg NaHPO, 300 mg chlorhexidine, mafenide, hexamethylpararosaniline, NaH2PO 25 mg aminacrine, phenazopyridine and ethoxazene: Purified water 9410 mg 15 ii) amines with antibiotic activity, selected from the group consisting of amikacin, gentamicin, kanamy cin, bekanamycin, neomycin, streptomycin, tobra mycin, lincomycin, clindamycin, erythromycin, EXAMPLE 3 colistin, polymyxin B, tetracycline, chlorotetracy Vaginal- cream 20 cline, rollitetracycline, oxytetracycline, spectinomy cin, viomycin, bacampicyline and stallimycin (A Composition for 100 g product Distamycine); iii) tromantadine with antiviral activity, BMA traumatate 1.75 g Glycolethylenaminophenol 12.2 g 25 iv) amines with antifungal activity, selected from the Propylenglycol 11.25 g group consisting of miconazole, econazole, chlormi White mineral jelly 6.5 g conazole, chlormidazole, isoconazole, bifonazole, Sodium cetyl stearyl sulphate 2.73 g Cetyl stearyl alcohol 19.57 g diamthazole, halethazole and hexetidine; and Decyl oleate 8.5 g d) a cation of a metal selected from silver and zinc. Methyl p-hydroxybenzoate 69 Ing 30 2. The traumatic acid salt as claimed in claim 1, wherein Propyl p-hydroxybenzoate 29 mg B is a quaternary ammonium cation selected from the group Purified water q.s. to 100 g consisting of: hexadecyltrimethylammonium, dodecyltrim ethylammonium and octyltrimethylammonium or a mixture thereof, benzyldimethylhexadecylammonium, benzyldim EXAMPLE 4 35 ethyldodecylammonium and benzyldimethyloctylammo nium or a mixture thereof; benzethonium, methylbenzetho Douche nium, cetylpyridinium, cetyldimethylammonium, Composition for 100 ml product dodecyldimethyl (2-phenoxyethyl)-ammonium and hexade cyl(2-hydroxychlorohexyl) dimethyl-ammonium. ETA traumatate 17.5 g 40 3. The traumatic acid salt as claimed in claim 1, wherein, Perfume 5 mg when B is a cation as defined in class (b), it is selected from Purified water q.s. to 100 ml the group consisting of ethanolamine and 2-propanolamine, diethanolamine and di-2-propanolamine. We claim: 4. A therapeutic composition for the therapeutic treatment 1. A trans and cis traumatic acid salt of formula (I) and (II) as of cutaneous pathologies when the bacteriostatic, antibiotic, antifungal or antiviral activity is associated to a cicatrizant Booc1N1\N1N1\-1S-COOB (D effect containing as the active ingredient at least one trans and/or cis traumatic acid salt of formula (I) and (II) Booc1N1N1N1 N1 N CooB (ID 50 Booc1N1,N1\-1N1 S-COOB (D wherein B is a cation as defined in one of the following classes: Booc1N1 N1\-1N1 N CooB (II) a) quaternary ammonium cations of formulae (III) or (IV) wherein B is a cation as defined in one of the following Re (III) 55 classes: R -N -R a) quaternary ammonium cations of formulae (III) or (IV) R4 R(*) (III) R(t) (IV) R --R, wherein R, R2, R and R are equal or different from each R(+) (IV) other and are selected from the group consisting of: i) a linear or branched alkyl radical having from 1 to 20 R- -R7 carbon atoms optionally containing in the aliphatic 65 wherein R, R, R and R are equal or different from chain at least one of the following groups: arylenoxy, each other and are selected from the group consisting alkylenoxy, and being optionally substituted in the of: 5,567,716 25 26 i) a linear or branched alkyl radical having from 1 to 20 d) a cation of a metal selected from silver and zinc in carbon atoms optionally containing in the aliphatic combination with suitable excipients and/or diluents. chain at least one of the following groups: arylenoxy, 5. The therapeutic composition as claimed in claim 4 for alkylenoxy, and being optionally substituted in the aliphatic chain with at least one of the following the treatment of injuries and infected injuries, fistulae, residues: aryl, arlyoxy and alkoxy groups; and necrotic processes, ulcerodystrophic alterations, acne, der ii) a cycloalkyl radical of from 3 to 10 carbon atoms; Rs matitis seborrheic, neurodermatitis, itches, dermic phenom and R in formula (IV) form with the nitrogen atom ena of intoxication, trachoma, herpetic keratitis and verru apyridine ring, R is a C-Clinear or branched alkyl C2C, radical; O 6. The therapeutic composition as claimed in claim 4, wherein B is a quaternary ammonium cation selected from b) a cation of linear or branched C-C mono-, di-, or the group consisting of hexadecyltrimethylammonium, tri-alkanolamine; dodecyltrimethylammonium and octyltrimethylammonium c) a cation of a primary, secondary or tertiary amine belonging to one of the following subclasses: or a mixture thereof, benzyldimethylhexadecylammonium, i) amines with disinfectant, antiseptic and bacterio 15 benzyldimethyldodecylammonium and benzyldimethyloc static activity, selected from the group consisting of tylammonium or a mixture thereof; benzethonium, methyl chlorhexidine, mafenide, hexamethylpararosaniline, benzethonium, cetylpyridinium, cetylidimethylammonium, aminacrine, phenazopyridine and ethoxazene, dodecyldimethyl (2-phenoxyethyl)-ammonium, hexade ii) amines with antibiotic activity, selected from the cyl(2-hydroxychlorohexyl) dimethyl-ammonium. group consisting of amikacin, gentamicin, kanamy 20 7. The therapeutic composition as claimed in claim 4 cin, bekanamycin, neomycin, streptomycin, tobra wherein, when B is a cation as defined in class (b) it is mycin, lincomycin, clindamycin, erythromycin, Selected from the group consisting of ethanolamine and colistin, polymyxin B, tetracycline, chlorotetracy 2-propanolamine, diethanolamine and di-2-propanolamine. cline, rollitetracycline, oxytetracycline, spectinomy 8. The therapeutic composition as claimed in claim 4, cin, viomycin, bacampicyline and stallimycin (A 25 administrable by parenteral route. Distamycine); 9. The therapeutic composition as claimed in claim 8, iii) tromantadine with antiviral activity, wherein the parenteral route is the intramuscular one. iv) amines with antifungal activity, selected from the 10. The therapeutic composition as claimed in claim 4, group consisting of miconazole, econazole, chlormi administrable by topical route. conazole, chlormidazole, isoconazole, bifonazole, 30 diamthazole, halethazole and hexetidine; and k k k k