CA2204145A1 - Acne treating-wound healing compositions containing a pyruvate, an antioxidant and a mixture of fatty acids - Google Patents

Abstract

This invention pertains to therapeutic acne treating-wound healing compositions useful for the topical treatment of acne vulgaris. The compositions comprise a therapeutically effective amount of tretinoin and a wound healing composition. In one embodiment the wound healing composition comprises (a) pyruvate; (b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids. The therapeutic acne treating-wound healing compositions may be utilized in a wide variety of pharmaceutical products. This invention also relates to methods for preparing and using the therapeutic acne treating-wound healing compositions and the pharmaceutical products in which the therapeutic compositions may be used. This invention also relates to methods for employing the therapeutic acne treating-wound healing compositions to treat wrinkles.

Description

ACNE TREATI~:C h~UND HEALING COMPOSITIONS CONTAINING A PYRUVATE, AN ANTIOXIDANT
AND A MIXTURE OF FATTY ACIDS

BACKGROUND OF THE INVENTION

1. Field of the Invention This invention pertains to therapeutic acne treating-wound healing compositions useful for the topical ll~ of acne vulgaris. More particularly, theacne treating-wound healing compositions comprise tretinoin and a wound healing composition andlor its metabolites. This invention also pertains to methods for p.~,..g and using the acne treating-wound healing compositions and the ph~rm~cel-tical products in which the therapeutic compositions may be used. Thisinvention also relates to meths)tl~ for employing the the.~c.l~ic acne treating-wound healing compositions to treat wrinkles.
A pl~r~-led embodiment of the therapeutic wound healing composition of this invention comprises (a) pyruvate selected from the group con~i~t ng of pyruvic acid, ph~ celltically acceptable salts of pyruvic acid, and llli~Lul~S thereof, (b) anantioxidant, and (c) a Il~ of saluralcid and ~m.~ d fatty acids whcl~ill the fatty acids are those fatty acids required for the repa* of cellular membranes and resuscitation of "~ ""~ n cells.

W O 96/14868 ~CTrUS95/12853 2. Description of the Background Wound Healing s Wounds are intemal or extemal bodily injuries or lesions caused by physical means, such as meçh~nic~l chemical viral, bact~ri~l or themlal means, which disrupt the nommal co..l;.ul;~y of structures. Such bodily injuries include collhlei~ n~, wounds in which the skin is unbroken, incisions, wounds in which the skin is broken by a cutting in~l~ul~ t~
10 and lacerations, wounds in which the skin is broken by a dull or blunt insl~u~ . Wounds may be caused by ~ccid~nt.~ or by surgical procedures. Patients who suffer major wounds could benefit from an ~nh~nc~-m~nt in the wound healing process.

Wound healing consists of a series of processes whereby injured tissue is repaired, speci~li7ecl tissue is regen~r~te~l, and new tissue is reor~ e(l Wound healing con~i~t~ of three major phases: a) an infl~mm~tion phase (0-3 days), b) a cellular proliferation phase (3-12 days), and (c) a r~m-.d~ling phase (3 days-6 months).

During the ;.. ll~,.. ;.l;on phase, platelet ag~l~g~lion and clotting form a matnx which traps plasma proteins and blood cells to induce the influx of various types of cells.
During the cellular proliferation phase, new connective or gr~nlll~t on tissue and blood vessels are formed. During the remodeling phase, gr~mll~tion tissue is replaced by a network of collagen and elastin fibers leading to the formation of scar tissue.

When cells are injured or killed as a result of a wound, a wound healing step is desirable to resll~cit~te the injured cells and produce new cells to replace the dead cells.
The healing process IC4UileS the reversal of cytotoxicity, the ~u~ ssion of infl~mm~tion, and the stimlll~tion of cellular viability and proliferation. Wounds require low levels of oxygen in the initial stages of healing to suppress oxidative damage and higher levels of oxygen in the later stages of healing to plolll~le collagen form~tion by fibroblasts.

W O96/14868 PCTrUS9S/128S3 M~mm~ n cells are continuously exposed to activated oxygen species such as superoxide (~2-)- hydrogen peroxide (H2O2), hydroxyl radical (OH-), and singlet oxygen (102). In vivo, these reactive oxygen ;--le~ *s are generated by cells inresponse to aerobic metabolism, catabolism of drugs and other xenobiotics, ultraviolet S and x-ray r~ tion, and the le~ilal~ y burst of phagocytic cells (such as white blood cells) to kill invading bacteria such as those introduced through wounds. Hydrogen peroxide, for example, is produced during re~ilalion of most living org~nicme especially by stressed and injured cells.

These active oxygen species can injure cells. An important example of such damage is lipid peroxidation which involves the oxidative degradation of unsa~ula~ed lipids. Lipid peroxi~l~tio~ is highly t~ to llwllll"~le structure and function and can cause IlUlllC;lUUS cytopathological effects. Cells defend against lipid peroxidation by producing radical scavengers such as superoxide ~licmnt~ce c~t~l~cç7 and peroxidase. Injured cells have a decreased ability to produce radical scavengers.
Excess hydrogen peroxide can react with DNA to cause backbone breakage, produce mutations, and alter and liberate bases. Hydrogen peroxide can also react with pyrimidines to open the 5, 6-double bond, which reaction inhibits the ability ofpyrimidines to hydrogen bond to compl~mt~nt~ry bases, ~ .nl~çr et al. (1971). Such oxidative biochernical injury can result in the loss of cellular lllel~ e integrity, reduced enzyme activity, changes in transport kinetics, changes in membrane lipid cont~ nt, and leakage of potassium ions, amino acids, and other cellular m~teri~l Antioxidants have been shown to inhibit damage associated with active oxygen species. For example, pyruvate and other Alpha-kçto~ci~ls have been reported to react rapidly and stoi~hiom~tically with hydrogen peroxide to protect cells from cytolytic effects, O'Donnell-Tormey et al., J. Exp. Med., 165, pp. 500-514 (1987).

United States Patents Nos. 3,920,835, 3,984,556, and 3,988,470, all issued to Van Scott et al., disclose methods for treating acne, dandruff, and palmar keratosis, respectively, which consist of applying to the affected area a topical composition comprising from about 1% to about 20% of a lower aliphatic compound co..~ lil.g from two to six carbon atoms selected from the group con.~i~ting of Alpha-WO 96tl4868 PCT/US95/12853 .hy~uAyacids, Alpha-ketc~ci(ls and esters thereûf, and 3-hyJluA~ullyic acid in aph~rm~r,ellhc~lly acceptable carrier. The ~lirh~hc comrolln~l~ include pyruvic acid and lactic acid.

United States Patents Nos. 4,105,783 and4,197,316, both issued to Yu et al., ~Close a method and composition, respectively, for treating dry skin which c~n~
of applying to the affected area a topical composition comprising from about 1% to about 20% of a co~ oul~d selecte~l from the group conci~ing of amides and allllll~lliUlll salts of Alpha-hydroxyacids"~-hydroxyacids, and Alpha-kçto~citle in a ph~rm~ce~lhcally acceptable carrier. The compounds include the amides and a~llolliul~l salts of pyruvic acid and lactic acid.

United States Patent No. 4,234,599, issued to Van Scott et al., fii~closes a method for treating actinic and non~r,tinic skin keratoses which consists of applying to the affected area a topical composition comprising an t;rrcc~ive amount of a col~ ,ul~d selected from the group con~i~hng of Alpha-hydroxyacids"~-hydroxyacids, and Alpha-ketoacids in a ph~rm~cellhcally acceptable carrier. The acidic compounds include pyruvic acid and lactic acid.

United States Patent No. 4,294,852, issued to wilrln~ r et al., tli~closes a composition for treating skin which comrri~es the Alpha-hydroxyacids"~-hydroxyacids, and Alpha-k~t~cids disclosed above by Van Scott et al. in combination with C3-C8~lirhs~tic alcohols.

United States Patent No. 4,663,166, issued to Veech, discloses an electrolyte sohlhl n which comrri~es a llliA~UlC of L-lactate and pyruvate in a ratio from 20:1 to 1:1, respectively, or a llliAlulc of D-~-hydl.)~yl,ulyl~tc and ~ceto~cePte in a ratio from 6:1 to 0.5:1, respectively.

Sodium pyruvate has been reported to reduce the number of erosions, ulcers, and h~morrh~ges on the gastric mucosa in guinea pigs and rats caused by acetylsalicylic acid. The analgesic and antipyretic properties of acetylsalicylic acid were not impaired by sodium ~yluv~le, Puscl~ -, Arzneimittçlforschung, 33, pp. 410415 and 415416 (1983).

Pyruvate has been reported to exert a positive inotropic effect in s~mned S myocdl~ ", which is a prolonged ~,~ .I. ;c~ r dysfunction following brief periods of coronary artery occlllcionc which does not produce i,.~ v~ ible ~l~m~ge Mçnt7~r et al., Ann. Surg., 209, pp. 629-633 (1989).

Pyruvate has been reported to produce a relative stabili~tiQn of left ventricular ~ s;,ule and work ~a~ . and to reduce the size of infarctions. Py~u~/~te ill,~lvves i~un.l,lion of spontaneous beating of the heart and re~ ;c)n of normal rates and pressure development, Bunger et al., J. Mol. Cell. Cardiol., 18, pp. 423438 (1986), Mochizuki et al., J. Physiol. (Paris), 76, pp. 805-812 (1980), Regitz et al., Cardiovasc. Res., 15, pp. 652-658 (1981), ~i~nnelli et al., Ann. Thorac. Surg., 21, pp. 386-396 (1976).

Sodium pyruvate has been reported to act as an ~nt~gonict to cyanide intoxication (p~ lably through the form~tion of a cyanohydrin) and to protect against the lethal effects of sodium sulfide and to retard the onset and development of fimction~l morphological, and biochemical measures of acrylamide neuropathy of axons, Schw~-Lc et al., Toxicol. Appl. Ph~ col.~ 50,pp. 437~42 (1979), Sabri et al., Brain Res., 483, pp. 1-11 (1989).

A chemotherapeutic cure of advanced L1210 lenk~mi~ has been reported using sodium pyruvate to restore abn-)rm~lly dçfo~m~d red blood cells to n~rm~l The d~r~lllled red blood cells plciv~ d adequate drug del*ery to tumor cel!s, Cohen,Cancer Chemother. Ph~rm~cQl, 5, pp. 175-179 (1981).

Primary cultures of h~L~.oL~pic tr~clle~l transplant exposed in vivo to 7, 12-dimethyl-benz(a)anthracene were reported to be snccessfully .. ~ ;.. e 1 in enrichment metlillm supple ~-~ ed with sodium ~yluvale along with cultures of interleukin-2stimnl~ted peripheral blood lymphocytes, and pl~em~cytomas and hybridom~, pig embryos, and human blastocysts, Sh~Ctpr~ J. Tmmllnol. Methods, 99, pp. 259-270 W O96/14868 PCTrUS95/12853 (1987), Marchok et al., Cancer Res., 37, pp. 1811-1821 (1977), Davis, J. Reprod. Fertil. Suppl., 33, pp. 115-124 (1985), OL~~ lo et al., No To Shinkei, 38, pp. 593-598 (1986), Cohen et al., J. In Vitro Fert. ~nLlyo Transfer, 2, pp. 59-64 (1985).
s United States Patents Nos. 4,158,057, 4,351,835, 4,415,576, and 4,645,764, all issued to Stanko, ~li.e~lose methode for pr~ lg the aGcnm~ hQn of fat in theliver of a I" .."".Al due to the ingestion of alcohol, for conhrolling weight in a ",~..""~l, for inhibiting body fat while increasing protein concPntr~hon in a ".i1",.,,~1 and for conhrolling the deposition of body fat in a living being, respectively. The methods comrriee ~lminiet~ring to the ",i1"~",~l a therapeutic l~ c; of pyruvate and dihydroxyacetone, and optionally riboflavin. United States Patent No. 4,548,937,issued to Stanko, discloses a method for controlling the weight gain of a ".i1."",~l which comprises a-lminietqring to the ."~"",~l a therapeutically effective amount of pyn~vate, and optionally riboflavin. United States Patent No. 4,812,479, issued to Stanko, fliecloses a method for controlling the weight gain of a .,.~.",,.~l which comprises a~lminietering to the ...,.~ 1 a therapeutically effective amount of dihydroxy~cetone and optionally riboflavin and pyruvate.

Rats fed a c~lcillm-oxalate lithogenic diet including sodium pyruvate were reported to develop fewer urinary calculi (stones) than control rats not given sodium yluval~, Ogawa et al., Hinyokika Kiyo, 32, pp. 1341-1347 (1986).

United States Patent No. 4,521,375, issued to Houlsby, ~lieclQses a methocl for sterili7ing sllrf~ces which come into contact wi~ living tissue. The method cQmrrie~,4 st.~rili7ing the surface with aqueous hydrogen peroxide and then neuhrali_ing the surface with pyruvic acid.

United States Patent No. 4,416,982, issued to Tauda et al., tliecloses a method for decomposing hydrogen peroxide by reacting the hydrogen peroxide with a phenol or aniline d~l;v~Live in the presence of peroxi~l~ee WO 96/14868 PCT/US9~/12853 United States Patent No. 4,696,917, issued to Lindstrom et al., discloses an eye irrig~tion solution which comrri~s Eagle's M;..;..., .., F.~nt-~l Me~lium with Earle's salts, chondroitin sulfate, a buffer solution, 2~ caylu~ nol~ and a pyruvate.
4 The irrigation solution may optionally contain ascorbic acid and Alpha-tocûpherol.
United States Patent No. 4,725,586, issued to Lindstrom et al., .li~CIQses an irrig~ n solution which ct mpn~es a balanced salt solution, chondroitin sulfate, a buffer solution, 2-~1cr~a~ ethanol, sodium bicarbonate or dextrose, a ~y~lvalt;, a sodium phosph~te buffer system, and cystine. The irri~tion solution may optionally contain ascorbic acid and gamma-tocopherol.
United States Patent No. 3,887,702 issued to Baldwin, discloses a composition for treating fingernails and toenails which consists ess~-nti~11y of soybean oil or sunflower oil in combination with Vitamin E.

United States Patent No. 4,847,069, issued to Bissett et al., discloses a phot~l~lective composition comprising (a) a sorbohyd,v~ -c acid, (b) an anti-infl~..,l,,~lolyagentselectedfromsteroidalanti-infl~ ..yagentsandanaturalanti-infl~.n".i.lnly agent, and (c) a topical carrier. Fatty acids may be present as an emollient. United States Patent No. 4,847,071, issued to Bissett et al., fli~closes a phot~ olective composition co.. ,~ ;.,g (a) a tocopherol or tocopherol ester radical scavenger, (b) an anti-infli3."",Atc,.y agent selected from steroidal anti-infl~ c.-y agents and a natural anti-infli3"...,~.ly agent, and (c) a topical carrier. United States Patent No. 4,847,072, issued to Bissett et al., discloses a topical composition comprising not more than 25% tocopherol sorbate in a topical carrier.
United States Patent No. 4,533,637, issued to Yamane et al., fli~closes a culture meflillm which comprises a carbon source, a nucleic acid source precursor, amino acids, vit~min~, miner~l~, a lipophilic nlltrient, and serum albumin, and cyclo~lPxtnn~ The lipophilic substances include unsaturated fatty acids and lipophilic vitamins such as Vitamin A, D, and E. Ascorbic acid may also be present.

United Kingdom patent applic~tion no. 2,1 96,348A, to Kovar et al., discloses a synthetic culture m~ m which comprises inorganic salts, monosaccharides, amino W O96/14868 . PCTAUS95/12853 acids, vit~min~, bu~.;ng agents, and optionally sodium ~y~lvale adding m~gn~ m hydroxide or magnesium oxide to the ~mlllc;on The oil phase may include chi~t-enfat.

United States Patent No. 4,284,630, issued to Yu et al., .li~closf~s a method for stabilizing a water-in-oil eml~ o7l which comprises adding m~gnf ~il7m hydroxide or magnesium oxide to the emulsion. The oil phase may include chi~7~5n fat.

Pl~.~,alation HTM has been reported to increase the rate of wound healing in artificially created rectal ulcers. The active ingredients in ~ ;on HlM are skinlGi~ l,y factor and shark liver oil, Subl~lyam et al., Digestive Diseases and Sciences, 29, pp. 829-832 (1984).

The addition of sodium ~y~llv~Le to b~cteri~l and yeast systems has been reported to inhibit hydrogen peroxide production, enhance growth, and protect the systems against the toxicity of reactive oxygen intfnnf~ tf,s The unsaturated fat~
acids and s~LulaL~d fatty acids contained within chicken fat enhanced me~ e repair and reduced eyL(~to~icity. The antioxidants ghlt~thione and thioglycollate reduced the injury inrluced by oxygen radical species, Martin, Ph.D. thesis, (1987-89).
United States Patent No. 4,615,697, issued to Robinson, discloses a controlled release Lle~l,..f .1 composition comprising a treating agent and a bioa~he~ive agent comrri~ing a water-swellable but water-insoluble, fibrous cross-linked carboxy-functional polymer.
European patent application no. 0410696Al, to Kellaway et al., (li~:closf.~s a mllcoa~lhesive delivery system COlll~lisillg a treating agent and a polyacrylic acid cross-linked with from about 1% to about 20% by weight of a polyhydroxy compound such as a sugar, cyclitol, or lower polyhydric alcohol.~0 Tretinoir.

Tretinoin (Retin-ATM) is in-lic~ted for the topical ll~ ~1... .1 of acne vulgaris.
Although the exact mode of action of tretinoin is unknown, it is believed that tretinoin S decreases the cohesiveness of follicular epith~ cells with decreased microcc)me<lo formation. Tretinoin also stimnl~tes mitotic activity and increases the ~ wv~L of folli~ r epithelial cells causing extrusion of the c-m~d~-nes Because tretinoin decreases the cohesiveness of follicular epithelial cells, tretinoin is also used to decrease wrinkles.
Topical use of tretinoin is known to cause the skin of some individuals to bccollle excessively red, edtom~tous blistered, or crusted. Topical use may also induce severe local erythema and peeling at the site of application. When these effects occur, the medication is either disco~ ed or the dosage lowered to a level the patient can tolerate. Topical use of tretinoin may also cause a height~.ned susceptibility to snnli,~ht as well as to weather extremes such as wind or cold. In ~d~ition, drug inter~ctioni with other topical medications such as abrasive soaps and cle~n~ers and soaps and cosmehcs having a strong dlying effect are also a concern.

Sl)MMARY OF THE INVENIION
This invention pertains to therapeutic acne treating-wound healing compositir~n~useful for the topical ~ of acne vulgaris. The compositions comprise a therapeutically effective amount of tretinoin and a wound healing composition and/or its metabolites. In one embodiment the wound healing composition comprises (a) yluv~le selected from the group con.~i~ting of pyruvic acid, ph~rm~ce~ltic~lly acceptable salts of pyruvic acid, and ~lul~s thereof; (b) an antioxidant; and (c) a fi~ e of saturated and nn~ d fat~y acids ~Lc.~ the fatty acids are those fatty acids required for the repair of cellular ~ lllbl~les and resll~cit~tion of l~ n~ 30 cells. The therapeutic acne treating-wound healing compositions may be utilized in a wide variety of ph~rm~ce~ltic~l products. This invention also relates to meth~ls for .al;llg and using the acne treating-wound healing therapeutic compositions and the ph~rm~centical products in which the therapeutic compositions may be used. This invention also relates to methods for employing the therapeu~.c acne treating-wound healing co~ o~i~ions to hreat wrinkles.

This invention further cornpriees allg,..~ d therapeutic acne hreating-wound S healing compositions c~.. l,. ;~;.. g acne treating agents and a the ~pc.ll;c wound healing composition in combination with one or more ~d~iihQn~l m~iic~...,~,.,l~i useful for treating wounds. This invention also relates to mP,th~le for preparing and using the ~ rnP.nted therapeutic acne treating-wound healing compositions and the ph~rm~ce~lhcal products in which the ~llgmPnted compositions may be used.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 depicts in bar graph format the viability of U937 monocytic cells following exposure of the cells to various ~nboxi~l~nte (Examples 1-5).
Figure 2 depicts in bar graph format the viability of U937 monocytic cells following exposure of the cells to various combin~h~n.~ of antioxidants (Examples 6-13).

Figure 3 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells following exposure of the cells to various ~ntir)xitl~nt~ (Examples 14-18).

Figure 4 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells following exposure of the cells to various combin~h~na of antioxidants (Examples 19-26).

Figure 5 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells following exposure of the cells to various combin~hon~ of ~nboxi~nti with and without a ~ of satulaled and unsalulated fatty acids (Examples 27-32).

WO 96/14868 . PCT/US95/12853 Figure 6 depicts in bar graph forrnat the levels of hydrogen peroxide produced by epidermal k~.~tinocytes following exposure of the cells to various ~nti~xid~ntc with and without a ~uld of s~hlr~ted and nnc~ ed fatty acids (Examples 33-42).
Figure 7 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal ker~tinocytes following exposure of the cells to various combinations of antioxidants with and without a ~ lule of saturated and uns~ d fatty acids (Examples 43-52).
Figure 8 depicts in bar graph format a s~ .y analysis of the levels of hydrogen peroxide produced by epidermal keratinocytes following exposure of the cells to the individual components of the wound healing composition, to various combinations of the wound healing composition, and to the wound healing lS composition.

Figure 9 is a photograph of wounded mice after 4 days of lle~ -lt with:
lion H (Example A); a petrolatum base fonnl~ n CQ~ live yeast cell d~;liv~Live, shark oil, and a llliX~ of sodium ~y,uv~l~, vitamin E, and çhick~n fat (Example B); a petrolatum base formlll~hon CQ~ ;ll;llg live yeast cell derivative and shark oil (Example C), and no composition (Example E, control).

Figure 10 is a photograph of a wounded mouse after 4 days of lle~l...~."
with a petrolatum base fomnll~hon only (Example D).

DETAILED DESCR~PTION OF THE INVENTION

This invention pertains to therapeutic acne treating-wound healing compositions which comprise tretinoin and a wound healing composition. ~ one embodiment the wound healing composition compri~es (a) py,uv~e sçlected from the group con~i~ting of pyruvic acid, ph~rm~ce~ltcally acceptable salts of pyruvic acid, and llli~ ,Sthereof; (b) an ~ntioxifl~nt; and (c) a ""x~ of saturated and unsaLul~aled fatty acids wherein the fat~ acids are those fatty acids required for the repair of cellular...~...l,.; .~es and r~cllcritAh~)n of ll~ AliAn cells.

Tretinoin is useful for the topical treAtm.~nt of acne vulgaris but is known to S induce excessive redness, e(l~mAtous blicterin~ or crusting, and several local erythema and peeling at the site of application. Wound healing compositions can increase the re,sllcc.itAtio~ rateof...i~ AliAn cellsandtheproliferationrateofnew...;~ .AliAn cells to replace dead cells but do not treat acne vulgaris. Applicant has found that the combination of tretinoin and a wound healing composition results in a therAre~1tic acne treating-wound healing composition which reduces the duration and sevelily of acne vulgaris and the irritation associated with tretinoin. This i Iv~,nlio.~ also relates to mt~.tho/l.c for employ-ing the therapeutic acne treating-wound healing composihonc to treat wrinkles.

Applicant has discov~;led therapeutic wound healing compositions for p,e~ ti~lg and reducing injury to ~ " "~ n cells and increasing the resn~cit~tion rate of injured ll~i1l""-~ n cells. Cells treated with the therapeutic wound healing compositions of this invention show decreased levels of hydrogen peroxide pro-lnction, increased resistance to cytotoxic agents, increased rates of proliferation, and increased viability. Cellular cultures c.)l.l;l;.. ;.. g the therapeutic wound healing compositions showed enhanced dirr~ iation and proliferation over control cultures and rapidlyformed ~tt~ lrlll~ or tight junctions between the cells to form an epidermal sheet.
Wounded ...~ treated with the therapeutic wound healing compositions show si~ific~rltly i~ r~v~d wound closing and healing over llllLI~.ated ".;.."".~lc and .- i -.. ~l~ treated with collvr~.l;on~l healing compo~ition~ The wound healingcomposition~ may be used alone or in combination with other m~ic~

The therapeutic wound healing compositions of this invention are described as Embodiment One. There are several aspects of Embodiment One. In a first aspect, (I.A), the therapeutic sound healing composition comrri~es (a) ~yluvale selected from the group con~i~tinE of pyruvic acid, phArm~çelltiç~lly acceptable salts of pyruvic acid, and l~ ulc;s thereof, (b) an ~ntioxitl~nt, and (c) a ll~Lul~; of s~tllr~tecl and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of W O 96114868 PCTrUSg5/12853 cellular membranes and res~l~cit~tiQn of ~ n.~ n cells. In a second aspect of EmboAim~ont One a.B), the the.~c.llic wound healing composition comrri~es (a) uv~le selecteA from the group conci~ting of pyruvic acid, ph~rm~r,elltic~lly acceptable salts of pyruvic acid, and ~lfLxlulcs thereof, (b) lactate s~lected from the S group concict1ng of lactic acid, ph~rm~cel)t1r.~lly acceptable salts of lactic acid, and l~lwcs thereof, and (c) a l~luuc of s~ d and nn~ d fatty acids whOEein the fatty acids are those fatty acids required for the repair of cellular l~ l,l~,es and re~ cit~ti-m of ~ .l"~ n cells. In a third aspect of Embodiment One a.C), the therapeutic wound healing composition compri~s (a) an ~ntioxiA~nt and (b) a llfi~Lul'e of s~ ed and unsalul~l~d fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular lll~,llI)l~les and rçs..~cit~tion of ~"..""~ n cells.
In a fourth aspect of Embodiment One a.D), the therapeutic wound healing composition comprises (a) lactate selected from the group c~ ting of lactic acid, ph~rm~celltically acceptable salts of lactic acid, and ~lfi~ules thereof, (b) anantioxidant, and (c) a llli~UiC of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids l~lui~cd for the repair of cellular llwllll~ es and resuscitation of ll.~.",ll~ n cells.

The therapeutic wound healing compositions of this invention are further combined with tretinoin, used for the topical L~r,~ r~lt of acne wlgaris. The r~,snlt~nt therapeutic acne treating-wound healing composition reduces the Anr~hon and severity of acne vulgaris and the irrit~hon associated with tretinoin. This invention also relates to methods for employing the therapeutic acne hreating-wound healing compositions to treat wrinkles.

The therapeutic acne treating-wound healing compositions of this invention may be further combined with one or more ~dtlihon~l m~lic~m~nti for hreating wounds to form ~llgm~nte~l acne treating-wound healing composihon.c This invention also relates to methods for pl~ g and using the ~ ...r.l~;d therapeutic acne treating-wound healing compositions and the ph~rm~ceutical products in which the ~ngm-.nted compositions may be used.

W O96114868 PCTnUS95/12853 The term "iniured cell" as used herein means a cell that has any activity di~lu~LGd for any reason. For example, an injured cell may be a cell that has injured w~ es or damaged DNA, RNA, and ribosom~s, for example, a cell which has (a) injured ~ bl~les so that transport through the l,.G~ es is (liminiched reslllting in S an increase in toxins and normal cellular wastes inside the cell and a decrease in mltnentc and other components necçc.c~ry for cellular repair inside the cell, (b) an increase in concentr~tion of oxygen radicals inside the cell because of the decreased ability of the cell to produce ~ntimri~ntc and &~yllleS, or (c) d~m~ged DNA, RNA, and ribosomes which must be repaired or replaced before normal cellular functions can be resllmeA The term "res.lcçit~hon" of injured ~ n cells as used herein means the reversal of cytotoxicity, the stabili~ti~n of the cellular ll.e.lll,ld-le, an increase in the proliferation rate of the cell, and/or the norm~li7~ )n of cellular functions such as the secretion of growth factors, hormon~c, and the like. The term "cytotoxicity" as used herein means a condition caused by a cytotoxic agent that injures the cell.Injured cells do not proliferate because injured cells expend all energy on cellular repair. Aiding cellular repair pl~ O~s cellular proliferation.

The term "prodrug", as used herein, refers to compounds which undergo biotran~ l~tion prior to exhibiting their ph~ cological effects. The chemical modification of drugs to overcome ph~rm~ceutical problems has also been termed "drug l~tenti~tion-ll Drug l~tenh~hQn is the chemiç~l modification of a biologically active compound to form a new compound which upon in vivo enzymatic attack will liberate the parent compound. The ehemic~l alterations of the parent compound are such that the change in physicochemical pr~.,Lies will affect the absorption, di~llibuLion and enzymatic metabolism. The ~lefinihr)n of drug l~tt-.nh~hon has also been elrt~.n.l~l to include nonenzymatic reg~.nl-.r~ti-)n of the parent compound.
~egeneration takes place as a consequence of hydrolytic, dissociative, and otherre~cPon.c not necessarily enzyme ...~ The terms prodrugs, l~tenh~ted drugs, and biorc;versible derivatives are used interchangeably. By inference, l~tenh~hon implies a time lag element or time component involved in regen~r~hng the bioactive parent molecnle in vivo. The term prodrug is general in that it inclllcles l~tenti~tecl drug de~ivaLives as well as those substances which are conv~,L~d after ~dminichation to the actual substance which combines with lec~ . The term prodrug is a generic term for agents which undergo biotran~rullll~lion prior to exhibiting their pharmacological ~ctione. In the case where the aAmini~t~red drug is not the active agent, but rather is biotransformed to the active agent, the term "prodrug" also in~.hlAes co.ll~oullds which may not nece~rily undergo biotr~n~r~,."i";on to the a-lmini.~ttored drug but mayundergo biotran~r,lll.alion to the active agent which exhibits the desired ph~rm~cological effect.

The term "metabolite", as used herein, refers to any snbst~nce produced by metabolism or by a metabolic process. "Metabolism", as used herein, refers to the various chemic~l reactioni involved in the tran~""~l;on of molecules or chemicalcompounds occllrring in tissue and the cells therein.

I. Wound Healin~ Compositions A. Embodiment One (I.A-D) The cells which may be treated with the therapeutic wound healing compositions in the present invention are l"i."""~ n cells. ~lthollgh applicant will describe the present therapeutic wound healing compositions as useful for treating ~ """~ n epidermal ker~tinocytes and ~"i~""~ n monocytes, applicant co~ )lates that the therapeutic wound healing compositions may be used to protect or resuscitate all l"; """~ n cells. Keratinocytes are representative of normal 111; 1"",~ n cells and are the fastest proliferating cells in the body. The correlation between the reaction of ker~tinocytes to injury and therapy and that of " " " " "~ n cells in general is very high.
Monocytes are lc~l~s~in~~ e of speci~li7~?d ~";1~""~ n cells such as the white blood cells in the immune system and the organ cells in liver, kidney, heart, and brain. The ."i.""~ n cells may be treated in vivo and in vitro.

Epidermal keratinocytes are the speci~li7ed epithelial cells of the epidermi~
which synthe~i7~ keratin, a scl~ rvl~ill which is the principal con~titn~ont of epidermii, hair, nails, horny tissue, and the organic matrix of the enamel of teeth.
M~mm~ n epidermal keratinocytes con~itllte about 95% of the epidermal cells and together with melanocytes form the binary system of the epidPrmi~ In its various W O96/14868 PCTrUS95/12853 successive stages, epiderrnal keratinocytes are also known as basal cells, prickle cells, and gr~n~ r cells.

Monocytes are monc)nllcle~rphagocytic leukocytes which undergo le~ila~ly S bursting and are involved in reactive oxygen ~ ed damage within the ep~ rmic Leukocytes are white blood cells or corpuscles which may be classified into two main groups: ~r~nnl~r leukocytes (granulocytes) which are leukocytes with abundant Er~mllPs in the cytoplasm and nongranular leukocytes (non~mllocytes) which are leukocyteswithout specific granules in the cytoplasm and which include the lymphocytes andmonocytes. Phagocyte cells are cells which ingest microor~nicmc or other cells and foreign particles. Monocytes are also known as large monon~1cle~r leukocytes, and hyaline or hr~nCition~l leukocytes.

Epiderrnal keratinocytic cells and monocytic cells have multiple oxygen generating mech~ni.cm.c and the degree to which each type of mPch~ni.cm functions differs in each type of cell. In monocytes, for example, the rc~pi.~tcly bursting process is more pronounced than in epiderrnal keratinocytes. Hence, the components in the therapeutic wound healing compositions of the present invention may vaTy clepPn-ling upon the types of cells involved in the con~ibon being treated.
As set out above, in a first aspect of Embodiment One a.A), the ther~re~hc wound healing composition for hreating ", ~ n cells, preferably epidermal kt~r~hnocytes, comprises (a) pyruvate sçlecte~l from the group con~ichng of pyruvic acid, ph~rm~ce~lhc~lly acceptable salts of pyruvic acid, and llfi~lui~s thereof, (b) an ~ntioxi(1~nt, and (c) a ~ c; of s~tm~ted and uns~ ed fatty acids wherein the fatty acids are those fat~ acids required for the repair of cellular ~ es and resuscitation of ",i1.""~ n cells. In a second aspect of Embo.limPnt One a.B), the therapeutic wound healing comrosihon for treating ",;."""~ n cells, preferably epiderrnal keratinocytes, comrrices (a) py~uvd~e selected from the group concichng of pyruvic acid, ph~rm~ceuhc~lly acceptable salts of pyruvic acid, and ~ lules thereof, (b) lactate s-?lectecl from the group con~i~ting of lactic acid, ph~rm~cellhc~lly acceptable salts of lactic acid, and ~ s thereof, and (c) a n~ e of s~ ecl and nnc~ to(l fatty acids wherein the fatty acids are those fatty acids required for the WO 96/14868 . PCT/US95/12853 repair of cellul3~ membranes and reSllecit~tio~ of ~ """~ n cells. In a third aspect of EmboAimtont One (I.C), the thC.~ ;c wound healing comrositi~n for treating ~";..,.."~ n cells, preferably epidçrm~l ker~tinocytes, comrriees (a) an ~nti~xiA~nt and (b) a ~ lule of s~ ecl and lme~t~ t~ fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular ~e~ es and reSlleçit~tion of ."".Ali~n cells. In a fourth aspect of Embodiment One (I.D), the th.o~pe~ltlc wound healing composition for treating ".~."."~ n cells, preferably monocytes, cnmpriees (a) lactate selectecl from the group con.c;etinE of lactic acid, ph~rm~celltir.~lly acceptable salts of lactic acid, and ~ uleS thereof, (b) an antioxidant, and (c) a llli~Ule of ~kl.. lçd and unsalula~ed fatty acids wherein the fatty acids are those fatty acids r~quiled for the repair of cellular lllellll,l~les and reslleçit~tion of "".."",~ n cells.

Pyruvic acid (2-ox~o~anoic acid,~lpha-k~l~ionic acid, CH3COCOOH) or pyruvate is a fim(l~mPnt~l intt-rmçAi~te in protein and carbohydrate metabolism and in the citric acid cycle. The citric acid cycle (tricarboxylic acid cycle, Kreb's cycle) is the m--ajor reaction sequence which Pxçcntçs the reduction of oxygen to generate ad~noeine triphosphate (ATP) by oxilli7ing organic compounds in respiring tissues to provide electrons to the transport system. Acetyl coenzyme A ("active acetyl") is Q~riAi7çd in this process and is thel~eanel utilized in a variety of biological processes and is a precursor in the biosynthesis of many fatty acids and sterols. The two major sources of acetyl coenzyme A are derived from the metabolism of glucose and fatty acids. Glycolysis ccn.~ict~ of a series of tran~r ....~I;on~ wherein each glucose molecule is transformed in the cellular cytoplasm into two ml-lec~lles of pyruvic acid.
Pyruvic acid may then enter the mitochondria where it is oxiAi7çd by co~,~yl,le A in the presence of enzymes and cofactors to acetyl coenzyme A. Acetyl coenzyme A can then enter the citric acid cycle.

In mll~cle, pyruvic acid (derived from glycogen) can be reduced to lactic acid during anerobic metabolism which can occur during exercise. Lactic acid is reoxicli7ed and partially retr~n~iro~ A to glycogen during rest. Pyluvale can also act as anantioxidant to neutralize oxygen r~lic~l~ in the cell and can be used in the mllltifunction oxidase system to reverse ~;y~icity.

The p~Tuvate in the present invention may be selected from the group cr~nQiQ~ing of pyruvic acid, ph~m~cellh~ lly acceptable salts of pyruvic acid, prodrugs of pyruvic acid, and l~ lulcs thereo~ In gener~l, the ph~Tm~ce~lh~lly acceptable salts of pyruvic acid may be alkali salts and ~lk~line earth salts. Preferably, the pyruvate S is selecte(1 from the group conQ;Q~ing of pyruvic acid, lithium pyluv~le, sodium ~,uvale, potassium pyruvate, m~gn~qillm pyruvate, G~lcium pyruvate, zinc py~uv~L~, m~ng~nese pyruvate, methyl pyruvate, Alpha-ketoglutaric acid, and ~Lules thereof.
More preferably, the py.uv~le is selecte(l from the group of salts conqiq-hn~ of sodium pyruvate, pot~isillm pyruvate, m--agnesium pyruvate, calcium pyruvate, zinc pyruvate, m~ng~nese pyruvate, and the like, and ~luu'~S thereof. Most preferably, the pyruvate is sodium pyruvate.

The amount of pyruvate present in the therapeutic wound healing compositions of the present invention is a therapeutically effective amount. A
therapeutically effective amount of l~yluv~le is that a-m-ount of pyruvate necessary for the inventive composition to pncvellt and reduce injury to ~ n cells or increasethe resuscitation rate of injured l~ n cells. The exact amount of l~yluvale is amatter of pl~r~.~..ce subject to such factors as the type of con~ihon being treated as well as the other ingredients in the composition. In a plc~ ,d embo-limPnt pyruvate is present in the therapeutic wound healing composition in an amount from about 10%
to about 50%, preferably from about 20% to about 45%, and more preferably from about 25% to about 40%, by weight of the therapeutic wound healing composition.

L-Lactic acid ((S)-2-hydlu~y~lo~ oic acid, (+) AIpha-hyd~ y~ ,ionic acid, CH3CHOHCOOH) or lactate occurs in small quantities in the blood and musclefluid of ,..~t~....~le Lactic acid concenh~hon increases in muscle and blood after vigorous activity. Lactate is a coll.~ollent in the cellular feedbaclr m~ch~ni.em and inhibits the natural re~i.~loly bursting process of cells thereby suppressing the production of oxygen radicals.
The lactate in the present invention may be selectecl from the group coneieting of lactic acid, ph~rm~celltic~lly acceptable salts of lactic acid, prodrugs of lactic acid, and ~Lules thereof. In general, the ph~rm~ce~ltic~lly acceptable salts of W O96/14868 PCTnUS9SI12853 lactic acid may be alkali salts and ~lk~line ear~ salts. Preferably, the lactate is s~le~,t~l from the group coneichn~ of lactic acid, lithium lactate, sodium lactate, pot~csillm lactate, m~gr ~cinm lactate, c~lc;llm lactate, zinc lactate, m~ng~nese lactate, and the like, and llli~Lu~Gs thereo~ More preferably, the lactate is s~lçct~ from the S group concict ng of lactic acid, sodium lactate, pot~icinm lactate, m~sillm lactate, c~lcinm lactate, zinc lactate, m~ng~n~se lactate, and ll~lUIGS thereof. Most preferably, the lactate is lactic acid.

The amount of lactate present in the therapeutic wound healing compositions of the present invention is a theraFel~tic~lly Grre~;~ive amount. A th~r~relltic~lly effective amount of lactate is that amount of lactate necos,s~ry for the i~lV~ iVG
composition to prevent and reduce injury to ~ n cells or increase the rçsllcçit~tion rate of injured l~ n cells. For an ingestible composition, a therapeutically effective amount of lactate is that amount necessary to suppress the ~~ aLuly bursting process of white blood cells to protect and resuscitate the ~ --l-"~ n cells. In general, a therapeutically effective amount of lactate in an ingestible composition is from about 5 to about 10 times the amount of lactate normally found in serum. The exact amount of lactate is a matter of plGrGlGllce subject to such factors as the type of con-lition being treated as well as the other ingredients in the composition. In a ~ler~;llGd embodiment, lactate is present in the therapeutic wound healing composition in an amount from about 10% to about 50%, preferably from about 20% to about 45%, and more preferably from about 25% to about 40%, byweight of the therapeutic wound healing composition.

Antioxidants are substances which inhibit oxi~l~hon or ~u~ /ss reactions pl~ ed byoxygenorperoxides. ~nhi-~xi-i~nti, especially lipid-soluble ~nhox~ nt~
can be absorbed into the cellular membrane to neutralize oxygen radicals and thereby protect the ll,~lllI,l~le. The antioxidants useful in the present invention may be selected from the group con~i~ting of all forms of Vitamin A (retinol), all forms of Vitamin2 (3, 4-didehydlul~ lol), all forms of carotene such as Alpha-carotene, ~-carotene (beta"~-carotene), gamma-carotene, delta-carotene, all forms of Vitamin C
(D-ascorbic acid, ~ascorbic acid), all forms of tocopherol such as Vitamin E (Alpha-tocopherol, 3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltri-decyl)-2H-l-W O96/14868 PCTrUS95/12853 be.~7c~yran-6-ol), ~-tocophercl, gamma-tocopherol, delta-tocopherol, tocoquinone, tocotrienol, and Vitamin E esters which readily undergo hydrolysis to Vitamin E such as Vitamin E acetate and Vitamin E succinate, and ph~rm~celltir.~lly acceptable Vitamin E salts such as Vitamin E phosph~te, prodrugs of Vitamin A, carotene, S Vitamin C, and Vitamin E, ph~ ce11tic~11y acceptable salts of Vitamin A, cal~te,~e, Vitamin C, and Vitamin E, and the like, and llfi~lults thereo~ Preferably, the ~ntioxi~l~nt iS selected from the group of lipid-soluble antioxidants conQiQ~ing of Vitamin A, ,~-carotene, Vitamin E, Vitamin E acetate, and llfi~ S thereof. More preferably, the antioxidant is Vitamin E or Vitamin E acetate. Most preferably, the ~ntioxitl~nt is Vitamin E acetate.

The amount of ~nti-)xitl~nt present in the therapeutic wound healing compositions of the present invention is a therapeutically effective ~m-lmt. A
therapeutically ~;live amount of ~ntioxi-l~nt is that amount of ~ntioxi~l~nt necessary for the inventive composition to plC;v~ and reduce injury to ~ n cells or increase the resuscitation rate of injured "-; "~ n cells. The exact amount of antioxidant is a matter of plercle.lce subject to such factors as the type of con-libon being treated as well as the other ingredients in the composition. In a ~ler~ d embo-lim~nt, the ~ntioxi~nt is present in the therapeutic wound healing composition in an amount from about 0.1% to about 40%, preferab1y from about 0.2% to about 30%, and more preferably from about 0.5% to about 20%, by weight of the therapeutic wound healing composition.

The ll~ ule of salulal~d and uns~ ed fatty acids in the present i~ nli.,n are those fatty acids required for the repair of "",."",~ n cellular ~ es and the production of new cells. Fatty acids are carboxylic acid compounds found in animal and vegetable fat and oil. Fatty acids are classified as lipids and are composed of chains of alkyl groups c~ ;..;..g from 4 to 22 carbon atoms and 0-3 double bonds and ch~r~ct~rized by a t~rmin~l carboxyl group, -COOH. Fatty acids may be s."...i.led or uns~ led and may be solid, semisolid, or liquid. The most CO11111l1J11 s~r~ted fatty acids are butyric acid (C4), lauric acid (C12), p~lmi~ic acid (C16), and stearic acid (Cl8). Unsaturated fatty acids are usually derived from vegetables and consist of alkyl chains co..l;t;..it~g from 16 to 22 carbon atoms and 0-3 double bonds with the W O96/14868 PCTrUS95/12853 characteristic tt-rmin~l carboxyl group. The most common unsaturated fatty acids are oleic acid, linoleic acid, and linolenic acid (all C18 acids).

In general, the llfLxlule of ~ and un~t~lr~ted fatty acids required for S the repair of ~ n cellular lllc~l,~es in the present illv~llLiun may be derived from animal and vegetable fats and waxes, prodrugs of sa~ d and nn~;al~ fatty acids useful in the present invention, and ~UI~S thereof. For example, the fattyacids in the therapeutic wound healing composition may be in the form of mono-, di-, or trigylcerides, or free fatty acids, or ~Lul~s thereof, which are readily available for the repair of injured cells. Cells produce the chemical components and the energy required for cellular viability and store excess energy in the form of fat. Fat is adipose tissue stored between organs of the body to furnish a reserve supply of energy. The plefe.led animal fats and waxes have a fatty acid composition similar to that of human fat and the fat contained in human breast milk. The pl~r~lled animal fats and waxes may be selecte.l from the group con.ciQ~ing of human fat, chirl~Pn fat, cow fat (defined herein as a bovine domt?~hc animal regardless of sex or age), sheep fat, horse fat, pig fat, and whale fat. The more plc;re.led animal fats and waxes may be stolecte(1 from the group co~ hng of human fat and chicken fat. The most plcf~ ;d animal fat is human fat. Mixtures of other fats and waxes, such as vegetable waxes (especiallysunflower oil), marine oils (especially shark liver oil), and synthetic waxes and oils, which have a fatty acid composition similar to that of animal fats and waxes, and preferably to that of human fats and waxes, may also be employed.

In a ~ref~llt;d embo-limPnt, the l~Lul~ of .~ t~d and lm~ ed fatty acids has a composition similar to that of human fat and compri~es the following fatty acids: butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, stearic, oleic acid, linoleic acid, linol~nic acid, ~r~l~hidic acid, and gadoleic acid. Preferably, butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, myristoleic acid, palmitic acid, p~lmit~leic acid, stearic, oleic acid, linoleic acid, linolenic acid, ~r~Chi~lic acid, and gadoleic acid are present in the ~lu-~ in about the following l,c~ellL~ges by weight7 respectively (carbon chain llulllbel and number of nn~ ;on~ are shown parenthetically, respectively): 0.2%-0.4% (C4), 0.1% (C6), 0.3%-0.8% (C8), 2.2%-3.5%

W O96/14868 PCTrUS95/12853 (Clo),0.9%-5.5% (Cl2)~2.8%-~.5% (c14),0.1%-0.6% (Cl4 l~,23.2%-24.6% (Cl6)~
1 8%-3 0% (C16 l),6.9%-9.9% (Cl8),36.0%-36.5% (Cl8 l)~20%-20.6% (Cl8 2)~
7-8% (C18:3)~ 1-1%_4-9% (C20), and 3.3%-6.4% (C20 1).

In another pLG~Ilcd embolim~nt, the ~~ e of Q,~ te<l and nne~
fatty acids is typically Ghir.1ren fat cQnlrricing the following fatty acids: lauric acid, myristic acid, myristoleic acid, p~nt~ec~n-1ic acid, p~lmitic acid, p~lmit(~leic acid, ic acid, l~gal~leic acid, stearic, oleic acid, linoleic acid, linol~nic acid, arachidic acid, and gadoleic acid. Preferably, lauric acid, myristic acid, myristoleic acid, p~.nt~-1ec.~noic acid, pa~itic acid, palmitoleic acid, m-argaric acid, ~ vleic acid, stearic, oleic acid, lino1eic acid, linolP.nic acid, arachidic acid, and gadoleic acid are present in the ~lfi~ e in about the following ~,..;cll~ges by weight, respectively:
( 12)' ~8% (Cl4)~0-2% (C14 1)~0-1% (Cl5),25.3% (Cl6),7.2% (C16 1),0.1%
(Cl7),0.1% (C17:1)'65% (C18)'37 7% (Cl8 l)~20-6% (Cl8 2),0.8% (Cl8 3),0.2%
lS (C20), and 0.3% (C20 l), all percentages +/- 10%.

In another plGrellGd embo-limPnt, the mixture of saLu~atcd and lme~hlr~te~l fatty acids comprieçs lecithin T.ecithin (phosphatidylcholine) is a phosphatide found in all living or~ni.em.e (plants and ~nim~l.e) and is a signifiç~nt coneh~ nt of nervous tissue and brain substance. Lecithin is a llfi~ c of the diglycerides of stearic, palmitic, and oleic acids, linked to the choline ester of phosphoric acid. The product of c~.. ~;.. ;e is pre(lomin~ntly soybean lecithin obtained as a by-product in the m~mlfz~ctllrin~ of soybean oil. Soybean lecithin cr~nt~ins palmitic acid 11.7%, stearic 4.0%, palmitoleic 8.6%, oleic 9.8%, linoleic 55.0%, linolenic 4.0%, C20 to C22 acids (includes ar~chitl/-nic) 5.5%. Lecithin _ay be represented by the formula:

CHOCOR
CH2O-P(O)2-OCH2CH2N (CH3)3 wherein R is se1ected from the group coneietin~ of stearic, p~1mibc, and oleic acid.

~ The above fatty acids and percentages thereof present in the fatty acid ul~ are given as an example. The exact type of fatty acid present in the fat~y acid G and the exact ~mollnt of fatty acid employed in the fatty acid ll~ . may be varied in ordOE to obtain the result desired in the final product and such v~ri~tione are S now within the capabilities of those skilled in the art without the need for undue CA~P~ lAl inn .

The amount of fatty acids present in the therapeutic wound healing compositions of the present invention is a therapeutically effective amount. A
therapeutically effective amount of fatty acids is that amount of fat~y acids neceec~Ty for the inventive composition to prGvGl.l and reduce injury to ",~.""~Ali~n cells or increase the resuscitation rate of injured In~ ,Ali~n cells. The exact amount of fatty acids employed is subject to such factors as the type and distribution of fatty acids employed in the ~ lule, the type of condition being treated, and the other ingredients in the composition. In a plGr~lled embodiment, the fatty acids are present in the therapeutic wound healing composition in an amount from about 10% to about S0%, preferably from about 20% to about 45%, and more preferably from about 25% to about 40%, by weight of the therapeutic wound healing composition.

In accord with the present invention, the therapeutic wound healing compoeitione of Embo-limt-nt One (I.A-D) for treating l~nlll",Ali~n cells may besçlected from the group conei~eting of:

a.A)(a) pyluvale s~lçcte(l from the group coneieting of pyruvic acid, ph~rrn~celltir~lly acceptable salts of pyruvic acid, and l~ LulGs thereof;
(b) an ~nhoxifl~nt and (c) a llli~Ule of s~tnr~ted and nnc~hlr~tçd fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular ~ "ll~ es and reS~lccit~tion of "~A."",~ n cells;
a.B)(a) pyruvate selected from the group concicting of pyruvic acid, ph~rm~rentir,~lly acceptable salts of pyruvic acid, and llfi~lules thereof;

(b) lactate selected from the group consisting of lactic acid, pharm~ce~lticallyaccept~hle salts of lactic acid, and ~ lul~.S thereof; and (c) a llPi~ of ~ d and nn~ l fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resllccit~t1on of S " .~" "" ,~ n cells;

aC) (a) an ~nt1oxi-l~nt and (b) a mi~lul~; of Si~ l~ and unsaLul~aled fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular ~ es and re,sn.ccit~ti. n of "~;."""~ n cells;

a.D) (a) lactate selected from the group c~ncictin~ of lactic acid, ph~rm~ce ltically acceptable salts of lactic acid, and l~ lUl~s thereof;
(b) an antioxidant; and lS (c) a llfi~lule of salul~led and unsalu~ted fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular mt-.rnkr~nes and resuscitation of ".;~ "l~ n cells.

Preferably, the wound healing compositions of Embo-lim~.nt One a) for treating ~ ""~ n cells, preferably epidermal keratinocytes, may be select~l from the group con~i~ting of:

(I.A) (a) ~yl uvale selecte~i from the group con.cicting of pyruvic acid, ph~rm~ce~lhcally acceptable salts of pyruvic acid, and llfi~lul~s thereof;
(b) an ~ntil~xitl~nt and (c) a ~ of s~hlr~ted and nn~c~tllr~te~ fatty acids wherein the fatty acids are those fatty acids requ*ed for the repa* of cellular mel~l~les and resuscitation of ",~."",~ n cells;

a.B) (a) pyruvate selected from the group coll.cichng of pyruvic acid, ph~rm~cenhc~lly acceptable salts of pyruvic acid, and llfi~lules thereof;
(b) lactate se1ect~d from the group concichng of lactic acid, ph~Tm~ceutically acceptable salts of lactic acid, and ll~lul~s thereof; and (c) a ~ xLu~e of s~ dted and unsaturated fatty acids wherein the fatty acids are those fatty acids lG.~ d for the repair of cellular .~ es and r~cn.crit~h-~n of " ,i.l "" ~ n cells; and ~.C) (a) an ~nhoxi-l~nt; and (b) a llfi~LWG of s~ ed and nnc~tllr~tsfl fatty acids where*l the fatty acids are those fatty acids required for the repa* of cellular ~ es and resl.cc.it~tion of ~ n cells.

10More preferably, the wound heal*ng compositions of Embo~liment One (I) for treating ~ n~ n cells, preferably epidermal keratinocytes, rnay be selecte<l from the group con.cichng of:

(I.A) (a) pyruvate selected from the group con~icting of pyruvic acid, 15ph~ cellhcally acceptable salts of pyruvic acid, and llf.~Lwes thereof;
(b) an ~nhoxill~nt; and (c) a n~i~LwG of s~ d and unsdLwaLGd fatty acids wherein the fatty acids are those fatty acids requ*ed for the repair of cellular IllG~ es and r~s-ccit~tion of,llillllll,~li~n cells; and (I.C) (a) an ~nhoxi~nt and (b) a mi~LulG of s~w~Gd and lm.c~tllr~ts-l fatty acids wherein the fatty acids are those fatty acids requ*ed for the repa* of cellular ~Gllll.l~les and resuscitation of l",,."",~ n cells.
More preferably, the wound healing compositions of Embo~lim~.nt One (I) for treating ~ n cells, preferably epicl~rn~l ker~tinocytes, may be st,lectscl from the group con~ichn~ of:

30~A) (a) pyruvate sçlecte~l from the group cnn~iC~ing of pyruvic acid, ph~rm~cel~hc~lly acceptable salts of pyruvic acid, and llf~Lw'eS thereQf;
(b) an ~nh-~xi-l~nt; and W O96/14868 . PCTrUS95/12853 (c) a llfi~LLule of saturated and unsaturated fatty acids wherein ~e fatty- acids are those fatty acids requ*ed for the repair of cellular m~ t-es and ~e;~..cc ;~ ;on of m~ l ll l lAliAn cells; and S (I.B) (a) pyruvate s~lecl~l from the group coneichng of pyruvic acid, ph~rmAce~lhc~lly acceptable salts of pyruvic acid, and llfi~lulGs thereof;
(b) lactate selectccl from the group concicting of lactic acid, ph~rmAcellt1cAlly acceptable salts of lactic acid, and llfi~lufGs thereof; and (c) a mi~Lule of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repa* of cellular membranes and resn.~citAtion of ~IlAllll~AliAn cells.

Most preferably, the wound healing compositions of Embodiment One (I) for treating IllAllllllAliAn cells, preferably epidermal keratinocytes, comprise:
(I.A) (a) ~yluvate selectecl from the group coll.cictin~ of pyruvic acid, ph~rrnAceutically acceptable salts of pyruvic acid, and llfi~lules thereof;
(b) an AntioxidAnt; and (c) a llli~lUl~; of s~hlr~ted and nncAtllr~ted fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular ll~ es and re,sll~citAtion of l"A~"",AliAn cells.

Most preferably, the wound healing compositions of Embodiment One (I) for treating IllA~ iAn cells, prefOEably monocytes, comprise:
(I.D) (a) lactate st-lecte~l from the group con~ictin~ of lactic acid, phArrn~cellticAlly acceptable salts of lactic acid, and llfi~ul~s thOEeof;
(b) an Ant oxi-lAnt and (c) a llf~x.Lule of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular llltlll~ es and r~,sllcçit~tion of mA-~ A1;An cells.

W O96/14868 PCTrUS95/12853 Throughout this disclosure, applicant will suggest various theories or ",P.r~ m~ by which applicant believes the cu~ ol,cnts in the therapeutic wound healing compositions and the antiviral agent fimchon together in an unexpected synergistic manner to plt;vellt and reduce injury to ~ n~ n cells, increase the reSll~çit~tion rate of injured ".~i""~Ali~n cells, and reduce viral titers. While applicant rnay offer various m~rh~nicm~ to explain the present invention, applicant does not wish to be bound by theory. These theories are suggested to better understand the present invention but are not inten~ed to limit the effective scope of the clairns.

In the first aspect of F.mbo limtont One a.A), applicant believes that pyluv~l~ can be transported inside a cell where it can act as an ~nti-xid~nt to nelltr~ e oxygen radicals in the cell. Pyruvate can also be used inside the cell in the citric acid cycle to provide energy to increase cellular viability, and as a precursor in the synthesis of important biomolecules to pr~,llote cellular proliferation. In ~ lition, ~y~uvale can be used in the multifunction oxidase system to reverse ~;y~,~icity. ~nhoxid~nt~,especially lipid-soluble antioxidants, can be absorbed into the cell ~ ane to neuhralize oxygen radicals and thereby protect the ,ll~."l,~ e. The sa~ulaled and unsaturated fatty acids in the present invention are those fatty acids required for the rçsll~cit~hon of ll,i1~"",~ n cells and are readily available for the repa* of injured cells and the proliferation of new cells. Cells injured by oxygen radicals need to produce unsalulalt;d fatty acids to repair cellular membranes However, the production ofm.c"~ tecl fatty acids by cells requires oxygen. Thus, the injured cell needs high levels of oxygen to produce lm~ cl fatty acids and at the same time needs to reduce the level of oxygen within the cell to reduce oxidative injury. By providing the cell with the unsaturated fatty acids needed for repair, the need of the cell for unsa~ul~led fatty acids is reduced and the need for high oxygen levels is also rednce~l The combination of ~y~u~/ale inside the cell and an ~nho~ nt in the cellular ~l~c;~ e functions in an unexpected synergistic manner to reduce hydrogen peroxide pr~d~1chon in the cell to levels lower than can be achieved by use of either type of component alone. The presence of ~lu,~s of saturated and unsahurated fatty acids in the therapeutic wound healing composition ~ignific~ntly enhances the ability of py~uva~ and the antioxidant to inhibit reactive oxygen production. By stabilizing the cellular membrane, unsaturated fatty acids also i~ ve Illclllblane function and enh~nce pyruvate transport into the cell. Hence, the three components in the IL~ crlLic wound healing composition of the first aspect of Embodim~nt One a~) ~unction together in an unexpected synergistic manner to ~lcvcnL and reduce injury to S l.. ,.. ~li~n cells and increase the resuscit~tion rate of injured .. ~.. n~ n cells.

In the second aspect of Embo~lim~nt One a.B), lactate is employed instead of an ~ntioxirl~nt Antioxidants react with, and neutralize, oxygen radicals after the radicals are already formed. T ~cPte, on the other hand, is a component in the cellular feedb~ m~ch~niem and inhibits the IC~i~ ly bursting process to ~u~)l~cSS
the production of active oxygen species. The collllJil.aLion of pyruvate to ne~ltr~li7e active oxygen species and lactate to su~less the l~,~il~loly bursting process functions in a synergistic manner to reduce hydrogen peroxide pro~luction in the cell to levels lower than can be achieved by use of either type of component alone. The presence of ll~Lulcs of saturated and unsaLu atcd fatty acids in the therapeutic wound healing composition significantly enhances the ability of pylllvale and lactate to inhibit reactive oxygen production. Hence, the three components in the therapeutic wound healing composition in the second aspect of Embo-lim~nt One a.B) function together in a synergistic manner to protect and reeuecit~te ll. .rl~ n cells.
In the third aspect of Embodiment One a.C), the presence of ll~ixLuues of saturated and unsaturated fatty acids in the therapeutic wound healing composition in this embodiment eignific~ntly ~ nh~n~es the ability of the ~ntioxil1~nt to inhibit reactive oxygen production. The combination of an ~ntioxirl~nt to neutralize active oxygen species and fatty acids to rebuild cellular lllc~ es and reduce the need of the cell for oxygen functions in a synergistic manner to reduce hydrogen peroxide pro~lnct on in the cell to levels lower than can be achieved by either type of culll~wlent alone. Hence, the components in the therapeutic wound healing composition in thethird aspect of Embodiment One a.C) function together in a synergistic manner toprotect and resnsc.it~te ~ n cells.

In the fourth aspect of Embo-lim~nt One a.D), lactate is employed because the rc~il~Lc~-y bursting process is more pronounced in monocytes than in W O96/14868 PCTrUS95/12853 epidermal keratinocytes. The combination of lactate to suppress ~e l~;laloly bursting process and an ~ntioxi-l~nt to n~ntr~ e active oxygen species functions in a synergistic manner to reduce hydrogen peroxide pro~ln(cti~n in the cell to levels lower than can be achieved by either cnmron~nt alone. The presence of l~ lul~,s of S s~lul~led and lm~ ecl fatty acids in the th~rentic wound healing comrositiQn in this emho-1im~nt ~ignific~ntly enhances the ability of lactate and the ~nti(~xi-l~nt to inhibit reactive oxygen pro~luction. Hence, the three col~ollents in the therapeutic wound healing composition in the fourth aspect of Embo-limt~nt One (I.D) fimct1-)n together in an unexpected synergistic manner to protect and res~lsçit~te ~ .."",~ n cells.

Accordingly, the combination of ingredients set out in the above embodime.ll~ fiunctions together in an enhanced manner to ~.~v~n~ and reduce injury to ll.~ n cells and increase the resll.~cit~tion rate of injured ",i~."",~lian cells.
The therapeutic effect of the combination of the components in each of the aboveemborli~ is m~rk~lly greater than that expected by the mere addition of the individual therapeutic components. Hence, applicant's therapeutic wound healing compositions for treating ",~"",.~ n cells have the ability to decrease intr~c.ell~ r levels of hydrogen peroxide proclllction, increase cellular r~si~t~nce to cytotoxic agents, increase rates of cellular proliferation, and increase cellular viability.

B. Methods For Making The Ther~pe~ Wound Healing Compositions Of Embodiment One (I.A-D) The present invention extends to methods for making the therapeutic wound healing composihons of F.mbo~im~-.nt One (I.A-D). In general, a therapeutic wound healing composition is made by forming an ad~ ure of the col~ol-ents of the composition. In a first aspect of Embodiment One (T.A), a therapeutic wound healing composition is made by forming an a.1..,;xl~.c of (a) ~yluvale selecte(l from the group con~;~tin~ of pyruvic acid, ph~rmace~ltiç~lly acceptable salts of pyruvic acid, and WO 96/14868 PCTtUS95/12853 m~Lules thereof, (b) an antioxidant, and (c) a llfi~lu~e of saturated and unsaturated fatty acids wherein the fatty acids are those fat~y acids required for the repair of cellular membranes and r~ c.it~tion of ~.".."".~ n cells. In a second aspect of Embodiment One ~.B), a therapeutic wound healing composition is made by forming S an ad,lf~lule of (a) ~yluvale s~lecte-l from the group crn~i~ting of pyruvic acid, ph~m~ce~1tic~11y acceptable salts of pyruvic acid, and llfi~lules thereof, (b) lactate sP1ected from the group con~icting of lactic acid, ph~ cel1tic~11y acceptable salts of lactic acid, and l~ Lul'~S thereof, and (c) a llli~luue of saturated and nn~ t~d fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular ~ alles and r~sllccit~t1on of ll~ n cells. In a third aspect of Embodiment One ~.C), a therapeutic wound healing composition is made by forming an ~ .;x~
of (a) an ~nh(!xid~nt and (b) a ~lul~. of s~ ed and nn~ ted fatty acids wherein the fatty acids are those fatty acids requ*ed for the repa* of cellular membranes and resuscitation of "li1-""~ n cells. In a fourth aspect of Embodiment One (I.D), alS therapeutic wound healing composition is made by forming an ~d-,,;xl---~ of (a) lactate s~1ected from the group con.~i~hng of lactic acid, ph~ ceutically acceptable salts of lactic acid, and l~ s thereof, (b) an antioxidant, and (c) a l~lule of salulaled and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular lllel~ es and rçsnscit~hon of "~..""~ n cells.
For some applications, the adl~lule may be formed in a solvent such as water, and a snrf~ct~nt may be added if required. If necessary, the pH of thesolvent is adjusted to a range from about 3.5 to about 8.0, and preferably from about 4.5 to about 7.5, and more preferably about 6.0 to about 7.4. The ~ ;x~ e is then sterile filtered. Other ingredients may also be inco~ ed into the therapeutic wound healing composition as dictated by the nature of the desired composition as well known by those having ordinary skill in the art. The n1tim~te therapeutic wound healing compo~ition~ are readily ~ ed using m~th~l~ generally known in the ph~ ce~1tic~1 arts.
In a ~ tll~d embodimlont, the invention is directed to a method for pl~alillg a therapeutic wound healing composition (I.A) for pl~v~ -lillg and redncing injury to m~mm~ n cells, and increasing the resuecit~tion rate of injured ~ n cells, which comrri.ees the steps of ~11mixing the following ingredients:
(a) pylu~dle s~lect~d from the group con.ei.eting of pyruvic acid, ph~rm~celltically acceptable salts of pyruvic acid, and llf~ ;s thereof;
S (b) an ~nhQxitl~nt and (c) a l~ of s~tnr~ted and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the reene~it~t1on of injured ~ n cells.

C. Methods For Employing The Ther~ql)e~ r Wound Healing Compositions Of Embodiment One ~ D) The present invention extends to methods for employing the therapeutic wound healing compositions of Embodiment One ~[) in vivo and in vitro. In general, a therapeutic wound healing composition is employed by cont~cting the therapeutic composition with .. -~.. ~li~n cells.

In a first aspect of Embodiment One (I.A), the invention is directed to a m~tho~ for plcvelltillg and redllcing injury to .. ;.. ~ n cells, and increasing the resuscitation rate of injured ~ n cells, which comprises the steps of (A) providing a therapeutic wound healing composition which comrrieçs (a) ~y~uval~
selectecl from the group con~ieting of pyruvic acid, ph~rm~ceuhc~lly acceptable salts of pyruvic acid,~and L~ ulcs thereof, (b) an ~nti~ nt, and (c) a l"~ e of e~ lçdand nne~tllr~tecl fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured " ~ "~ n cells, and (B) cont~chng the therapeutic woundhealing composition with the ~ n cells.

.
In a second aspect of Embodiment One (I.B), the invention is directed to a method for pleve~ g and reclllcing injury to ~- .;~ n cells, and increasing the resll.ecit~h~ n rate of injured ~ n cells, which comrriees the steps of (A) providing a therapeutic wound healing composition which comprises (a) pyruvate ~ ect~(1 from the group con.eiehng of py-ruvic acid,- ph~ cellhcally acceptable salts W O96/14868 PCT~US95/128S3 of pyruvic acid, and l~ ules thereof, (b) lactate selected from the group consisting of lactic acid, ph~rm~cellhc.~lly acceptable salts oftlactic acid, and ll~ s thereof, and (c) a l~ lul~ of sa~ula~d and nn~ fatty acids wherein the fatty acids are those fatty acids required for the rP,sll~cit~hon of injured ~ ."")~ n cells, and (B) cont~cting the therapeutic wound healing composition with the ,~ Ali~n cells.

In a third aspect of Embo-1imPnt One (I.C), the invention is dil~i~ led to a m~thod for pl~V~llLillg and re(lllcing injury to "~;..""~ n cells, and increasing the resuscitation rate of injured l"~l~""~ n cells, which comprises the steps of (A)providing atherapeutic woundhealing co",l,osi~ion which comrri~es (a) an antioxidant, and (b) a llli~Ule of s~ ed and lm~tllr~te(l fatty acids wherein the fatty acids are those fatty acids required for the resll~cit~tion of injured ,~ "",~ n cells, and (B) cont~chn~ the therapeutic wound healing composition with the ,~ n cells.

In a fourth aspect of Embodiment One ~[.D), the invention is directed to a method for preventing and reAllcing injury to ~n~ Ali~n cells, and increasing the rçsll~çit~tion rate of injured ~"i..""l~ n cells, which cc~mpri~Ps the steps of (A) providing a therapeutic wound healing composition which compri~P,s (a) lactate s~.lçcted from the group con~ ting of lactic acid, ph~rm~ce~ltic~lly acceptable salts of lactic acid, and l~ ules thereof, (b) an antioxidant, and (c) a llli~ule of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured ~ -"",~ n cells, and (B) cont~ctin~ the therapeutic wound healing composition with the "-;-""-~ n cells.

In a plcr~,led embo~imP.nt, the invention is directed to a method for healing a wound in a ",i.."",~l which c. mpri~es the steps of:
(A) providing a therapeutic wound healing composition (I.A) which comrri~i (a) pyruvate selected from the group con~i~ting of pyruvic acid, ph~rm~ce~hcally acceptable salts of pyruvic acid, and ~ules thereof;
(b) an ~ntioxitl~nt and (c) a ll~i~tule of s~tllr~ted and nn~tllr~ted fatty acids wherein the fatty acids are those fatty acids required for the resn~rit~tion of injured ~ n cells;and (B) contacting the therapeutic wound healing composition with the wound.

The types of wounds which may be healed using the wound healing compositions of Embodiment One (I.A-D) of the present invention are those which S result from an injury which causes epidermal damage such as incicion~ wounds in which the skin is broken by a cutting in~ , and l~c~ ti~ n.c, wounds in which the skin is broken by a dull or blunt in~ . The thcla~e~lLic composi~nc may also be used to treat various derm~tological disorders such as hypPrk~r~t~ic, photo-aging, burns, donor site wounds from skin transplants, ulcers (cutaneous, decubitus, venous stasis, and diabetic), psori~cic, skin rashes, and sunburn photoreactive procecces The topical therapeutic compositions may also be used orally in the form of a mouth wash or spray to protect and accelerate the healing of injured oral tissue such as mouth sores and burns. The topical therapeutic compositions may further be used in ophth~lmtlogical y~ dlions to treat wounds such as those which result from corneal ulcers, ra~ lk~loto~ly, corneal transplants, epikeratophakia and other surgically in~ ced wounds in the eye. The topical therapeutic compositions may in addition be used in anorectal creams and suppositories to treat such c~n~litionc as pruritus ani, proctitis, anal fissures, and hemorrhoids. In a ple~e.led embodiment, the therapeutic compositione are used to treat wounds such as inci~ion~ and l~ct-,r~ti->n~
The wound healing co~lyo~ilions of Embodiment One (I.A-D) of the present invention may be utilized in topical products, ingestible products, and tissue culture m~lillm to protect "I~""~ n cells and increase the resnccit~tion rate ofinjured ~ n cells. For example, the therapeutic wound healing composition~
may be used in topical skin care products to protect and increase the resn~çit~tion rate of skin tissue such as in the ll~ of various clerm~t~logical disorders such as hyp~lk.,.a~usis, photo-aging, and sunbuTn photoreactive processes. Injury to skin can occur for a variety of reasons. Injury often occurs to individuals who wash their hands often, to individuals who are exposed to stressful ~.lvilu~ 1 con~lihon~
(u~ .osnre to sun or chem-e~l~), or to the elderly or individuals with an lmdçrlining e The ~ddihQn of the wound healing compositions of the present invention to a lotion provides a source of ~nhoxi~1~nt~ to the skin which would protect the skin from the harrnful effects of W light, ~hemic~, and severe drying. The wound CA 0220414~ 1997-04-30 W O96/14868 PCT~US95/12853 healing compositions can be used for the following indications: a) Moisturizing and ;..g; b) T~çaling dry cracked skin; c) Treating ;..;~ ecl skin such as diaper rash, d) ~ealing severe dry skin due to other tlicç~çs (venous d~ ".~ ;C); e) Treatingpsori~i.c and other hyperproliferative ~1ice~cçs; f) Protecting skin from UV light S t1amage (antioxidant skin repl~cPm~.nt), g) Treating seborrheic con~iti-~nc, and h) Treating shaving wounds in an after shave lotion.

The topical therapeutic wound healing compositions may also be used orally in the form of a mouth wash or spray to protect and accelerate the healing of injured oral tissue such as mouth sores and burns. The topical therapeutic woundhealing compositions may further be used in ophth~lTn-~logical prep~ ionc such as eye care products to nentrali7e hydrogen peroxide used in the cleaning of contact lenses.
The topical therapeutic wound healing compositions may in acldition be used in anorectal creams and suppositnries to treat such conditions as pruritus ani, proctitis, anal fissures, and h~-molThoi~1c Initially as white blood cells enter a wound site, the cells release oxygen radicals, depleting the antioxidants at the wound site, thus ai~ g the healing process. Incc~l~uld~illg the wound healing compositions of thepresent invention into a wound healing form~ tiQn would f~-~.ilit~t~ healing by providing the site with usable ~ntioxitlantc, and a source of fatty acids needed for I~ lbl~le repair. The wound healing compositions can be used for the following indications: a) ~e~ling of cuts and scrapes; b) Burns (heals burns with less scaring and scabbing); c) Decubitus ulcers; d) Bed sores, E~res~uL~ ulcers; e) Fissures, Hemorrhoids;
f) Use in combination with immllnoshmnl~t~rs (.cimnlat~cl healing in healing deficient people); g) Post surgical wounds; h) R~nll~ges; i) Diabetic ulcers; j) Venous ulceration;
and k) Use in combination with wound cl~ncing agents.

The therapeutic wound healing compositions may also be used in ingestible products to protect and increase the resn~cit~tiQn rate of erosions, stom~c~
ulcers, and hemorrhages in the gastric mncos~ Other ingestible therapeutic products in~ lç stroke m~ ationc; ~u~ e disease mç~lic~tion~; ar~ritis mçdic~tlon~;
ulcerm~lic~ )n~; cancermedications (~iyL~ ic agents); heartm~lic~tion to h~ ve regional ventricular function and restore normal heart rate and plt;;s~ule functions; lung medication to repair injured tissue; liver m~lic~tiQn to sU~pl~Ss lipogenesis of alcoholic origin and prevent hepatic steatosis; kidney medication to suppress urina~ calculi (kidney stones); ~letoxifiç~tiQn m~lic~tlon to ~nt~goni7~g heavy metal poiconing, cyanide poi-C~ning, sodium sulfide p;)jRQll;~ , other types of pOicoll;llg ; and reduce and neutralize the pro~hlchon of oxygen r~-lic~lc which produces injury to tissue, to protect and further ~.nh~nce the resllc~it~tic)n rate of the injured "".~""~ n cells. The therapeutic wound healing compositions may be used in ingestible products to treat infl~ ce~ces such as hepat1tic, g~.etr~ti.c, colitis, esophagitis, arthritis, and pancreatitis.

The therapeutic wound healing compositions of the present invention may also be used in tissue culture media and organ transplant media to P1GVt;111 and reduce injury to ~ ."ll,~ n cells and increase the resuscitation rate of injuredn cells. Tissue cultures and transplant organs encounter reactive oxygen species generated in the culture media by the injured cells. Organs particularlysusceptible to oxidative damage during transport and transpl~nt~hc)n due to reperfusion injury following i.cc.hemi~ are corneas, livers, hearts, and kidneys. The therapeutic wound healing compositions may be useful to abrogate reperfusion injury to such transplant organs.

In a specific embo~lim~nt, the invention is directed to a method for preserving ~ n cells in a culture m~ ]m which comprises the steps of:
(A) providing a therapeutic wound healing composition selecte(l from the group of concicting Of (I.A) (a) ~y-uv~l~ selected from the group concicting of pyruvic acid, ph~rm~ce~lhc~lly acceptable salts of pyruvic acid, and ll~lul~;s thereof;
(b) an ~nhox~ nt and (c) a llfi~Lule of s~hlr~te~l and nn.c~ r~tçd fahy acids wherein the fatty acids are those fatty acids required for the repair of cellular l..r~..b.,1r~es and resuscitation of m~.""~ n cells;

(I.B) (a) pyruvate sçlectecl from the group conciching of pyruvic acid, ph~rm~celltically acceptable salts of pyruvic acid, and ~fi~lulc;s thereof;

W O 96/14868 . PCTrUS95112853 (b) lactate selected from the group consisting of lactic acid, ph~rm~sellt1c~11y acceptable salts of lactic acid, and ~lu,~s ~ereof; and (c) a lllU Lulc of 6.~ ted and nn~ d fatty acids vvll~cill the fatty acids are those fatty acids required for the repair of cellular ~ es and S rsSuccit~~ion of ~ n cells;

ac) (a) an ~ntioX~ nt and (b) a ~Lwe of c~t~ t~d and nn~llll i1t~l fatty acids wl.ert;in the fatty acids are those fatty acids required for the repair of cellular l,lcu.I"a,les and r~suscit~hon of ~ n cells;

a.D) ~(a) lactate selected from the group cona~ ng of lactic acid, ph~rm~centically acceptable salts of lactic acid, and nli~LIules thereof;
(b) an antio7ci-1~nt and (c) a ll iA~we of salulaled and unsalulal~d fatty acids wh~ the fatty acids are those fatty acids required for the repair of cellular lllc~ es and resn.ccit~tic-n of ,~ "",~ n cells; and (b) an ~ntioxi(l~nt; and (c) a ll~tU e of s~ 1 and unsatNrated fatty acids wherein the fatty acids are those fatty acids re~luilcd for the resll.ccit~tion of injured ~ "~ n cells;
(B) providing ll-~-llll,~li~n cells in a culture medilln~; and (C) c. nt~ting the ther~pel)tic wound healing composition from step (A) with the ~n cells in the culture meflillm from step (B).

D. Formulations Of The Therapeutic Wound Healing Compositions Of Embodiment One (I.A-D) Once prepared, the illvclltive therapeutic wound healing compositions of Embodiment One (I.A-D) may be stored for future use or may be formlll~ted in c~clivc ~l~wlls with ph~rm~ce~ltic~lly acceptable carriers to prepare a wide variety of pharmaceutical compositions. Examples of ph~rm~ceutically acceptable carriers are ph~rm~cellti~l appliances, topical vehicles (non-oral and oral), and inEP~tible vehicles.

Examples of ph~rm~ce~ltic~l appliances are sutures, staples, gauze, b~n~1~ges, burn dreeeinge, artificial skins, liposome or micell fot mnl~tir~ne microcapsules, aqueous vehicles for so~king gauze dreeeings~ and the like, and llf~LuleS
thereof. Non-oral topical compositions employ non-oral topical vehicles, such ascreams, gels form~ ti~n.e foams, o;.~ and sprays, salves, and films, which are intPnded to be applied to the skin or body cavity and are not in~Pn-le~l to be taken by mouth. Oral topical compositions employ oral vehicles, such as l~luuLhwashes, rinses, oral sprays, suspen~ions, and dental gels, which are intP.n~ed to be taken by mouth but are not inten-1P~d to be ingested. Tngestikle compoeitiorle employ ingestible or partly ingestible vehicles such as conrecl;Qn~ry bulking agents which include hard and soft collfe~;l;rnçry such as lozenges, tablets, toffees, nougats, suspen~i~ne, chewy c~n~ies, and chewing gums.

In one form of the invention, the therapeutic wound healing composition is incorporated into a ph~rm~ce~ltir~l appliance which may be in the form of sutures, staples, gauze, bandages, burn dreseinge, artificial skins, liposome or micell fs~rmlll~tio~e, microc~psllles, aqueous vehicles for so~king gauze dre~eeing~e~ and the like, and Illi~lules thereof. A variety of tr~liti~n~l ingredients may optionally be inr,]nded in the ph~rm~cellttcal composition in effective ~l.ull~ such as buffers, ~[~ lV~tiVtS, tonicity adjusting agents, antioxidants, polymers for adjusting viscosity or for use as PYtpn~lprs~ and excipients, and the like. Specific illustrative examples of such tr~tlition~l ingredients include acetate and borate buffOEs; ~ ,.osal, sorbic acid, methyl and propyl paraben and chlorobutanol preservatives; sodium chloride and sugars to adjust the tonicity; and excipients such as ...~ l, lactose and sucrose. Other cc,llv~ n~l ph~rm~ceutical additives known to those having ordinary skill in theph~rm~cellhc~l arts may also be used in the ph~rm~cellhc~l composition.

In accor~ ce with this invention, therapeutically effective ~IllUUllki of the therapeutic wound healing compositions of the present invention may be employed in the pharmaceutical appliance. These amounts are readily determined by those skilled in the art without the need for undue ~ e ;...~ ;OI- The exact ~OUnl of the therapeutic wound healing composition employed is subject to such factors as the type and conc~nh~h.~n of the therapeutic wound healing composition and the type of S ph~rm~celltic~l appliance employed. Thus, the amount of therapeutic wound healing composition may be varied in order to obtain the result desired in the final product and such variations are within the capabilities of those skilled in the art without the need for undue cA~ t;Qn In a p-~r~lled embotlimt-nt, the ph~rm~ce~lhc~l composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 5%, by weight of the ph~rm~rellhc~l composition. In a more ~l~relled embo-lim~nt, the ph~rm~cellhc~l composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 3%, by weight of the ph~rm~cel-tical composition. In a most ~iefell~,d embc!-1im~-nt, the ph~rm~cellhc~l composition will comprise the therapeutic wound healing composition in an amountlS from about 0.1% to about 1%, by weight of the ph~rm~ce~lhcal composition.

The present invention extends to methods for making the ph~rm~cellhc~l compositions. In general, a ph~rm~cellhc~l composition is made by contacting a therapeutically erre~i~ive amount of a therapeutic wound healing composition with a ph~rm~cel-tical appliance and the other ingredients of the final desired ph~rm~ce -hc~l composition. The therapeutic wound healing composition may be in a solvent and may be absorbed onto a ph~rm~cellhc~l appliance.

Other ingredients will usually be incorporated into the composition as tlict~ted by the nature of the desired cum~o~i~ion as well known by those havingordinary skill in the art. The nltim~te ph~rm~ce~lhc~l cull~o~ilions are readilyprepared using methods generally known in the ph~rm~cellhc~l arts.

In another form of the invention, the therapeutic wound healing composition is incorporated into a non-oral topical vehicle which may be in the form of a cream, gel, foam, ointm.-nt, spray, and the like. Typical non-toxic non-oral topical vehicles known in the ph~rm~ceutical arts may be used in the present invention. The pl~rt;lled non-oral topical vehicles are water and ph~ cellhc~lly acceptable water-miscible organic solvents such as ethyl alcohol, isopropyl alcohol, propylene glycol, glycerin, and the like, and ~ Ul~s of these solvents. Water-alcohol ~ ul'es are particularly pl~r~ d and are generally employed in a weight ratio from about 1:1 to about 20:1, preferably from about 3:1 to about 20:1, and most preferably from about 3:1 to about 10:1, ~eclively.

The non-oral topical therapeutic wound healing compositions may also contain cullv~,l.lional additives employed in those products. Convention~l additives include h~lm~ct~nt~, emollients, lubricants, stabilizers, dyes, and perfumes, providing the additives do not illlt;,r~le with the therapeutic properties of the therapeutic wound healing composition.

Suitable hnm~ct~nt.c useful in the non-oral topical therapeutic wound healing compositions include glycerin, propylene glycol, polyethylene glycol, sorbitan, fructose, and the like, and ~ es thereof. ~llmt?ct~nt~, when employed, may be present in al~ from about 10% to about 20%, by weight of the topica1 therapeuticwound healing composition.

The coloring agents (colors, colorants) useful in the non-oral topical therapeutic wound healing composition are used in amounts ~rre~;live to produce the desired color. These coloring agents include pi m~-nt~ which may be incorporated in alllUUlll:j Up to about 6% by weight of the non-oral topical therapeutic wound healing composition. A ~.~;rt;..~;d pigment, ~ 1 dioxide, may be incorporated in up to about 2%, and preferably less than about 1%, by weight of the non-oral topical therapeutic wound healing composition. The coloring agents may also include natural food colors and dyes suitable for food, drug and cosmetic applications. These coloring agents are known as F.D.& C. dyes and lakes. The m~teri~l~ acceptable for the foregoing uses are preferably water-soluble. Tlln~tr~tive nonlimiting examples include the indigoid dye known as F.D.& C. Blue No.2, which is the cli~o-linm salt of 5,5-indigotin(li~llfonic acid. Similarly, the dye known as F.D.& C. Green No.l c.~mpri~çs a triphenylmPth~ne dye and is the mnnrso-linm salt of 4-[4-(N-ethyl-p-sulro~ l)enzylamino) diphenylmethylene]-[l-(N-ethyl-N-~-sulruniulllbenzyl)-delta-2,5-cyclohP~ 1ito.neimine]. A full reçit~tion of all F.D.& C. coloring agents and their W O96/14868 PCTrUS95/12853 corresponding chemical structures may be found in the Kirk-Othmer Encyclopedia of ~h~ l Technology, 3rd FAiticm, in volume S at pages 857-884, which text is incul~u,aled herein by reference.

In accordance with this invention, therapelltiç~lly c;r~c~ivc ~uu~ of the therapeutic wound healing compositions of the present invention may be ~1mixç~1 with a non-oral topical vehicle to form a topical th~.,. ~c wound healing composition. These a~ull~ are readily ~letPTminçcl by those skilled in the art without the need for undue expc. ;.~ ;on In a plcr~ d embo-limPnt the non-oral topical therapeutic wound healing compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 10% and a non-oral topical vehicle in a quantity snffici~nt to bring the total amount of composition to 100%, by weight of the non-oral topical therapeutic wound healing composition. In a more pl'~r~ d embo-limPnt, the non-oral topical therapeutic wound healing compositions will comprise the therapeutic wound healing cc~ o~i~ion in an amount from about 0.1% to about 5%, and in a most pl~crcllcd embotlimPnt, the non-oral topical therapeutic wound healing compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and a non-oral topical vehicle in a quantity s~lfficient to bring the total amount of composition to 100%, by weight of the non-oral topical therapeutic wound healing composition.

The present invention extends to methods for prepalillg the non-oral topical therapeutic wound healing co~ o~ilions. In such a method, the non-oral topical therapeutic wound healing composition is prepared by ~tlmixing a therape~h~lly errcclive amount of the therapeutic wound healing composition of the present invention and a non-oral topical vehicle. The final compositions are readily pr~alcd usingstandard mPtho~l~ and apparatus g~ne~lly known by those skilled in the ph~Tm~ce~lhs~l arts. The app~lus useful in accordance with the present invention comrri~P,s mixing a~alus well known in the ph~rm~sel~lic~l arts, and therefore the selection of the specific ~pp~lus will be apparent to the aTtisan.

In another form of the invention, the therapeutic wound healing composition is incorporated into an oral topical vehicle which may be in the forrn of a ~ ulllw~sh~ rinse, oral spray, suspension, dental gel, and the like. Typical non-toxic oral vehicles known in the ph~rm~celltic~l arts may be used in the present invention.
The ~l~f~,l.ed oral vehicles are water, eth~nt l, and water-ethanol ~lul~s. The water-v ethanol l~fi~Lu es are generally employed in a weight ratio from about 1:1 to about 20:1, preferably from about 3:1 to about 20:1, and most preferably from about 3:1 to r about 10:1, respectively. The pH value of the oral vehicle is generally from about 4 to about 7, and preferably from about 5 to about 6.5. An oral topical vehicle having a pH value below about 4 is generally irrit~ting to the oral cavity and an oral vehicle having a pH value greater than about 7 generally results in an unpleasant mouth feel.

The oral topical thc.~ lic wound healing compositi~n~ may also contain convt-ntiorl~l additives norm~lly employed in those products. Convention~l additives include a fluorine providing compound, a ~wciete ~ g agent, a flavoring agent, a coloring agent, a hnm~ct~nt, a buffer, and an em~ er, providing the additives do not i,l~e,rt;,e with the therapeuhc properties of the therapeuhc wound healing composihon.

The coloring agents and l" - - - -rc~ , and the ~ I I II -U ~ of these addihves to be employed, set out above as useful in the non-oral topical therapeuhc woundhealing composihon may be used in the oral topical therapeutic wound healing composihon.

Fluorine providing compounds may be fillly or slightly water soluble and are characteri~ed by their ability to release fluoricle ions or fl~ori-le c~ g ions in water and by their lack of reaction with other components in the composition. Typical fluorine providing co...~vu lds are inorganic fluoride salts such as water-soluble alkali metal, alkaline earth metal, and heavy metal salts, for example, sodium fl~ori~le, pot~sillm fl~lori~le, ~ I l ln~ n~ flllori~le~ cuprous fll10ri(1e, zinc fluoride, stannic fllloricle, stannous flllori~le, barium flnori~le, sodium fluorosilic~te, ~,.. -. I-iU.. , fluorosilic~te sodiumfluo-o~ ate, sodiumm-noflllorophosph~te, ~l~l,n;~ .mono-and di-fluorophosphates and flllorin~tecl sodium c~lcillm pyrophosph~te Alkali metal fluorides, tin fluoride and monofluorophosphates, such as sodium and stannous fluoride, sodium monoflll~rophosph~tf, and ~lulGs thereof, are pl'GrGll~d.

The amount of fluorine providing compound present in the present oral S topical therapeutic wound healing compositinn is dependent upon the type of flnnrine providing compound employed, the solubility of the flllorine compound, and the nature of the final oral therapeutic wound healing composition. The amount of fluorine providing compound used must be a nnntoxic amount. In general, the flll~rine providing compound when used will be present in an amount up to about 1%, preferably from about 0.001% to about 0.1%, and most preferably from about 0.001%
to about 0.05%, by weight of the oral topical therapeutic wound healing composition.

When sweelf-.i.~g agents (~w~G~..r,rs) are used, those sweeteners well known in the art, including both natural and artificial ~weGL~ ers, may be employed.
The sweetP.ning agent used may be self,ct~-l from a wide range of m~tfri~l~ inclll-ling water-soluble sweetening agents, water-soluble artificial sweetening agents, water-soluble sweetf.ning agents derived from naturally occurring water-soluble sweete.ning agents, dipeptide based ~we~le~ g agents, and protein based sweet~nin~ agents, inCl~ nE ~ ules thereof. Without being limited to particular ~we~ .;..g agents, representative categories and examples include:
(a) water-soluble sweetf ning agents such as monos~cch~ri(les, cch~ritles and polysaccharides such as xylose, ribose, glucose (dextrose), m~nnose, g~l~ctose fructose (levulose), sucrose (sugar), m~lt~se invert sugar (a ~lule of~uctose and glucose derived from sucrose), partially hydrolyzed starch, corn syrup solids, dihydro~h~lcones, mnnellin, steviosides, and gly~;y~lhi;Gill, and ~ es thereof;
(b) water-soluble artificial sweeteners such as soluble s~cch~rin salts, i e., sodium or calcium .s~cch~rin salts, cyclamate salts, the sodium, ~lllllOlliUlll or c~lcium saltof3,4-dihydro-6-methyl-1,2,3-o~r~thi~ine-4-one-2,2~1ioxitle, thepot~iinm salt of 3,4-dihydro-6-methyl-1,2,3-ox~ .;..e-4-one-2,2-dioxide (Aces~1lf~m~-K), the free acid form of s~cch~rin, and the like;
(c) dipeptide based sweeteners, such as L-aspartic acid derived sweeteners, such as L-aspartyl-L-phenyla1anine methyl ester (Aspartame) and m~teri~
described in United States Patent No. 3,492,131, L-Alpha-aspartyl-N-(2,2,4,4-CA 0220414~ 1997-04-30 tetramethyl-3-thietanyl)-D-alanin-amide hydrate (Alitame), methyl esters of L-aspartyl-L-pheny1glycerineandL-aspartyl-L-2,5-dihydrophenyl-glycine, L-aspartyl-2,5 dihydL-phenyl~l~nine; L-aspartyl-L-(1-cyclohçYen)-alanine, and the like;
(d) water-soluble sweeteners derived from naturally occnrring water-soluble sweeteners, such as clllorin~ted d~liv~Lives of or~ uy sugar (sucrose), e.g., chlorodeoxysugar d~;valives such as d~;l;v~Lives of chlorodeoxysucrose or chlorodeoxyg~l~ct~s~lerose, known, for example, under the product ~lexign~ti~n of Sucralose; examples of chlorodeoxysucrose and chlorodeoxygalacto-sucrose del;v~Lives include but are not limited to: l-chloro-1'-deoxysucrose; 4-chloro4-deoxy-Alpha-D-galacto-pyranosyl-Alpha-D-fru~iL~ru.,.l-osi~le, or 4-chloro4-deoxyg~l~ct~s~ .rose; 4-chloro4-deoxy-Alpha-D-galacto-pyranosyl-l-chloro-1-deoxy-B-D-fructo-furanoside, or 4,1'-dichloro-4,1'-dideoxy~l~r,t~s lcrose; 1',6'~ichloro-1',6'-dideoxysucrose; 4-ch~oro-4-deoxy-Alpha-D-galacto-pyranosyl- 1 ,6-dichloro- 1 ,6-dideoxy-B-D-fructo-furanoside, or 4,1 ',6'-trichloro-4, 1 ',6'-trideoxygalacto-sucrose; 4,6-dichloro4,6-dideoxy-Alpha-D-galacto-pyranosyl-6-chloro-6-deoxy-B-D-frucLo~.~oside or 4,6,6'-trichloro4,6,6'-trideoxyg~l~ctosucrose; 6,1',6'-trichloro-6,1',6'-trideoxysucrose; 4,6-dichloro4,6-dideoxy-Alpha-D-galacto-pyranosyl-1,6-dichloro-1,6-di-deoxy-B-D-frucLorulalloside, or 4,6,1 l,6'-tetrachloro4,6, 1 ',6'-tetradeoxygalacto-sucrose; and 4,6,1 ',6'-tetrachloro-4,6,1',6'-tetradeoxy-sucrose; and (e) protein based sweetener~ such as th~llm~r~ccous danielli I and II).

In general, an effective amount of sweetening agent is utilized to provide the level of sweetness desired in the particular oral topical therapeutic wound healing composition, and this amount will vary with the sweetener st-lecte-l and the final oral therapeutic product desired. The amount of ~we~ er norm~lly present is in the range *om about 0.0025% to about 90%, by weight of the oral topical therapeutic wound healing composition, depending upon the sweetener used. The exact range of ~ tX
for each type of sweetener is well known in the art and is not the subject of the present invention.

The flavoring agents (flavors, flavol~lt~) which may be used include those flavors known to the skilled artisan, such as natural and artificial flavors.

Suitable flavoring agents include mints, such as peppermint, citrus flavors such as orange and lemon, artificial vanilla, cinn~mon, various fruit flavors, both individual and mixed, and the like.

S The amount of n~vO~ g agent employed in the oral topical th~ ,. lic wound healing composition is normally a matter of ElrGrGlGllce subject to such factors as the type of final oral therapeutic wound healing comrosition~ the individual flavor employed, and the strength of flavor desired. Thus, the amount of flavoring may be varied in order to obtain the result desired in the final product and such v~ri~t1on~ are within the capabilities of those skilled in the art without the need for undue ~1 c ;.n~ ;Qn The flavoring agents, when used, are generally utilized in alllUUllk~
that may, for example, range in ~lluullL~ from about 0.05% to about 6%, by weight of the oral topical therapeutic wound healing composition.

Suitable buffer solutions useful in ~e non-oral topical therapeutic wound healing compositions include citric acid-sodium citrate solution, phosphoric acid-sodium phosphate solution, and acetic acid-sodium acetate solution in alllUUllki Up to about 1%, and preferably from about 0.05% to about 0.5% by weight of the oral topical therapeutic wound healing composition.
In accordance with this invention, therapeutically effective a~ of the therapeutic wound healing compositions of the present invention may be ~mi~ced with an oral topical vehicle to form a topical therapeutic wound healing composition.
These ~llou~lLs are readily ~letprminp~d by those skilled in the art without the need for undue e,~c.;.. Pnt~1ion In a plerGllGd emb~limPnt, the oral topical therapeutic wound healing compositions will comrri~e the the.~ ic wound healing composition in an amount from about 0.1% to about 10% and a oral topical vehicle in a quantity snffi~iPnt to bring the total amount of composition to 100%, by weight of the oral topical therapeutic wound healing composition. In a more ~IGrGllGd embo-1imPnt, the oral topical therapeutic wound healing compositions will comprise the th~r~rel tic wound healing composition in an amount from about 0.1% to about 5%, and in a most plerGllGd embo.limPnt, the oral topical thGl~e.llic wound healing compositions will comprise the th~relltic wound healing comrosition in an amount from about 0.1%

W O 96/14868 PCTnUS9S/12853 to about 2%, and a oral topical vehicle in a quantity sufficient to bring the total amount of co~ osilion to 100%, by weight of the oral topical tk~ .e..l;c wound healing Collll.o~ilion.
r S The present invention extends to m~ th~s for pl~alillg the oral topical therapeutic wound healing compositions. In such a m. tho~, the oral topical therapeutic wound healing composition is prepared by ~lmi~ing a therapelltiç~lly ~rreetive ~ t of the ~erapeutic wound healing composition of the present invention and an oraltopical vehicle. The final compositions are readily ~r~a~ed using standard methods and apl~alus generally known by those skilled in the ph~rm~ce~ltic~l arts. The a~p~lus useful in accordance with the present invention comrriees mixing apparatus well known in the ph~rm~ce~lti~l arts, and therefore the se1ection of the specific a~p~tus will be ~ppaLc;llt to the artisan.

~ a plc;rel~ed embo-limçnt, an oral topical therapeutic wound healing composition is made by first dissolving coloring agents, sweetening agents, and similar additives in water. The therapeutic wound healing composition is then ~imixçd with the aqueous solution. Then sllfficient water or ethanol, or l~ luL~,S of water and eth~nol, are added to the solution with mixing until the final solution volume is reached. In a more ~-er~,.ed embo~im~nt the therapeutic wound healing composition is added to the solution as the final ingredient. The final oral topical therapeutic wound healing compositions are readily l,f~paLed using methods generally known in the ph~rm~centical arts.

The oral therapeutic wound healing composition may also be in the form of dental gel. As used herein, the term "gel" means a solid or semisolid colloid which cont~ine con~i~lPrable qn~nhtiÇs of water. The colloid particles in a gel are linked together in a coherent llRsllwolk which immobilizes the water co~ cd inside the . meshwork.
The dental gel compositions of the present invention may contain the cullv~ ;on~l additives set out above for oral topical therapeutic wound healing compositions such as ll~ulllwashes, rinses, oral sprays, and suspensions and, in W O96/14868 PCTrUS95/12853 addition, may contain additional additives such as a polishing agent, a ~lesçn~iti7ing agent, and the like, providing the ~ 1it1~n~l additives do not i~ r~.c with the d~c.llic properties of the t~ Lic wound healing cc,lll~,osilion.

In a dental gel composition, the ora1 vehicle genera11y COlll~l;SCS water, typically in an amount from about 10% to about 90%, by weight of the dental gel composition. Polyethylene glycol, propylene glycol, glycerin, and .. i~ ;S thereof may also be present in the vehicle as hnmect~nt~ or binders in ~ from about 18% to about 30%, by weight of the dental gel composition. Particularly prcrc-lcd oral vehicles comprise llfi~lu-cs of water with polyethylene glycol or water with glycerin and polypropylene glycol.

The dental gels of the present invention include a gelling agent (thickening agent) such as a natural or synthetic gum or gelatin. Gelling agents such as hydroxyethyl cellulose, methyl c~ 10se7 glycerin, carboxypolymethylene, and gelatin and dle like, and llfi~ cs thereof may be used. The pl~;rt;~cd gelling agent is hydroxyethyl ce11-11Ose Gelling agents may be used in ~~ lL~ from about 0.5%
to about 5%, and preferably from about 0.5% to about 2%, by weight of the dental gel composition.
The dental gel compositions of the present invention may also include a polishing agent. In clear gels, a po1i~hing agent of colloidal silica and/or aLkali metal ah~minosi1ic~te complexes is plcrt;"ed since these m~t~i~l~ have refractive indices close to the refractive indices of the gelling systems c~ ly used in dental gels.
In non-clear gels, a polishing agent of c~lsi11m carbonate or c~lci11m dihydrate may be used. These polishing agents may be used in amounts up to about 75%, and preferably in ~lluull~ up to about 50%, by weight of the dental gel composition.

The dental gel may also contain a ~lesen~i~7ing agent such as a collll",.~lion of citric acid and sodium citrate. Citric acid may be used in an amount from about 0.1% to about 3%, and preferably from about 0.2% to about 1%, by weight, and sodium citrate may be used in an amount from about 0.3% to about 9%, and preferably from about 0.6% to about 3%, by weight of the dental gel composition.

W O96/14868 pcTrus95ll2853 In accordance with this invention, therapeutically effective amounts of the therapeutic wound healing compositions of the present invention may be ~(lmixed into the dental gel compositicn~ These ~lluullL~ are readily ~let~ninçd by thoseskilled in the art without the need for undue experimt~nt~tion In a prc;rc;lled emboflim~nt the dental gel compositions will compri~e the th~l~e.llic wound healing composition in an amount from about 0.1% to about 10% and an oral topical vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight of the dental gel composition. In a more pl~ d embo-lim~nt, the dental gel compositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about S%, and in a most p.~rell~d embo-limtnt, the dental gelcompositions will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and an oral topical vehicle in a quantity sllffici~nt to bring the total amount of composition to 100%, by weight of the dental gel composition.
lS
The present invention extends to methods for preparing the therapeutic dental gel compositions. In such a meth~ the dental gel composition is prepared by ~tlmixing a therapeutically effective amount of the therapeutic wound healing colll~o~ilion of the present invention and an oral topical vehicle. The final compositions are readily prepared using m~thorls generally known by those skilled in the dental and ph~rm~cel~tical arts. The apparatus useful in accordance with thepresent invention comprises mixing a~pa~Lus well known in the ph~rm~celltic~l arts, and therefore the selection of the specific app~uaLus will be apparellt to the artisan.

2S In a pl~;rtll~d embo~lim~nt a therapeutic dental gel composition is made by first dispersing a gelling agent in a hllmect~nt or water, or a ~ lule of both, then admixing to the dispersion an aqueous solution ûf the water-soluble additives such as the fluorine providing compound, sweet~n~s and the like, then adding the polishing agent, and lastly admixing the flavoring agent and the therapeutic wound healingcomposition. The final gel ~ Luueis then tubed or otherwise packaged. The liquids and solids in a gel product are proportioned to form a creamy or gelled mass which is extrudable from a plc~ e~ container or from a collapsible tube. The final therapeutic wound healing compositions are readily prepared using methods generallv known in the ph~rm~ee~ti~l arts.

In yet another form of the invention, the therapeutic wound healing composition is incorporated into an ing~et1hle vehicle. The ingestible vehicle may be a collr~-;lionery bulking agent in the form of lozenges, tablets, toffees, nougats, suspenqion.e, chewy c~n~ies, chewing gums, and the like. The ph~rm~centi~lly acceptable carriers may be plblJalbd from a wide range of materials inclll-lin~, but not limited to, tlilll~ntq, binders and adhesives, lubricants"liqintt?grants, coloring agents, bulking agents, flavoring agents, sweetening agents and miqct~ neous m~teri~lq such as buffers and adso~ that may be needed in order to prGpar~ a particular therapeutic confection.

The preparation of cullfec! ;on~y formnl~tions is historically well known and has changed liffle through the years. Confectiont Ty items have been cl~qqifi~d as either "hard" confectionery or "soft" confectionçry. The th~ ulic wound healing compositions of the present invention can be incorporated into confection~y compositions by a~lmixing the il~velllive composition into col,v~ n~l hard and soft c.,llr~ .,lls.
As used herein, the term c~ fb.;l ;onr~ m~t~i~l means a product con~ ;..g a bulking agent selected from a wide variety of materials such as sugar, corn syrup, and in the case of s~lg~rl~qq bulking agents, sugar alcohols such as sorbitol and ~ ol and l~lul~s thereof. Confecti~ne~y m~teri~l may include such exemplary substances as lozenges, tablets, toffee, nougat, suspen.qionq, chewy candy, chewing gum and the like. The bulking agent is present in a ~lu~llily sllffici~nt to bring the total amount of composition to 100%. In general, the bulking agent will be present in alllOUllki Up to about 99.98%, preferably in ~~ up to about 99.9%, and more preferably in ~ up to about 99%, by weight of the ingestible therapeutic wound healing composition.

T~ npes are navored m~lic~ted dosage forms inten-le(l to be sucked and held in the mouth. Lozenges may be in the form of various shapes such as flat, circular, octagonal and biconvex forms. The lozenge bases are generally in two forms:
hard boiled candy lo~P.nges and cul~ es3cd tablet lozenges.

Hard boiled candy lozenges may be processed and form~ ted by col.v~.L;Qn~l means. In general, a hard boiled candy lozenge has a base composed of a ~ lulc of sugar and other carbohydrate bulking agents kept in an amorphous or glassy c~ nl1iti~n This amorphous or glassy form is crnQid~red a solid syrup of sugars ~en~r~lly having from about 0.5% to about 1.5% moisture. Such m~teri~lQ. nl~rm~lly contain up to about 92% com syrup, up to about 55% sugar and from about 0.1% to about 5% water, by weight of the final composition. The syrup component is gP.ner~lly p~ared from corn syrups high in fructose, but may include other m~teri~lQ Further ingredients such as flavoring agents, sweetening agents, ~ciclnl~nt~, coloring agents and the like may also be added.

Boiled candy lozenges may also be prepared from non-fermt-nt~kle sugars such as sorbitol, ~ ol, and hydrogenated corn syrup. Typical hydrogenatedcorn syrups are Lycasin, a commercially available product m~nnf~ctllred by Roquette Col~o~lion, and Hystar, a coll"l~clclally available product m~nllf~ctllred by Lonza, Inc.
The candy lozenges may contain up to about 95% sorbitol, a ll~ Ul~ of sorbitol and ... ~ ol in a ratio from about 9.5:0.5 up to about 7.5:2.5, and hydrogenated corn syrup up to about 55%, by weight of the solid syrup component.

Boiled candy lozenges may be routinely prepared by convention~l methotls such as those involving fire cook~r~, vacuum cookers, and scraped-surface cookers also referred to as high speed ~tmosphPric cookers.

Fire cookers involve the tr~dition~l method of making a boiled candy lozenge base. In this m~tho-l, the desired quantity of carbohydrate bulking agent is dissolved in water by heating the agent in a kettle until the bulking agent dissolves.
Additional bulking agent may then be added and cooking conhml~d until a final elalwe of 145~C. to 156~C. is achieved. The batch is then cooled and worked as a plastic-like mass to incorporate additives such as flavors, colorants and the like.

~ A high-speed atmospheric cooker uses a heat-exchanger surface which involves spreading a film of candy on a heat eyrh~nge sllrface~ the candy is heated to 165~C. to 170~C. in a few ..~;~u~l~s The candy is then rapidly cooled to 100~C. to 120~C. and worked as a plastic-like rnass enabling incorporation of the additives, such S as flavors, colorants and the like.

In vacuum cookers, the carbohydrate bulking agent is boiled to 125~C.
to 132~C., vacuum is applied and ~ it on~l water is boiled off without extra hr~hng When cooking is complete, the mass is a semi-solid and has a plastic-like con~i~t~ncy.
At this point, flavors, colorants, and other additives are ~lmixçd in the mass by routine m~chanic~l mixing opera~ion~

The opLilllulll mixing required to ullirollnly mix the flavoring agents, coloring agents and other additives during conven~on~l m~mlf~ctllring of boiled candy lS lozenges is det~-rmined by the time needed to obtain a ~---;rvl-ll distribution of the m~trri~l~ Normally, mixing times of from 4 to 10 ~ çs have been found to be acceptable.

Once the boiled candy lozenge has been pLop~ly tempered, it may be cut into workable por~ions or formed into desired shapes. A variety of forming techniques may be utilized depending upon the shape and size of the final product desired. A general discussion of the composition and ~ Lion of hard conre~Lions may be found in H.A. Lieberman, Ph~rm~ce~l~ical Dosa~e Forms: Tablets, Volume I
(1980), Marcel Dekker, Inc., New York, N.Y. at pages 339 to 469, which ~ closllre is incorporated herein by reference.

The ~Lus useful in accoldal~ce with the present invention c~ lises cooking and mixing apparatus well known in the confectionery m~nnf~r,tllring aTts, and therefore the selection of the specific ~ua~us will be apparent to the artisan.
In contrast, co.~ essed tablet confections contain particulate m~teri~l~
and are formed into st~uctures under ~JleSi~ulC. These confections gen~lly contain sugars in ~lloull~ up to about 95%, by weight of the composition, and typical tablet excipients such as binders and lubricants as well as n~vOl;llg agents, coloring agents and the like.

- - In ~ hnn to hard confectinn~ry m~t~ri~l~, the lozenges of the present invention may be made of soft confection~y m~t~n~le such as those conlai,led in nougat. The ~ aLioll of soft conre-iions, such as nougat, involves collve~lL;nn~l m~ho le, such as the combination of two primary components, namely (1) a high boiling syrup such as a corn syrup, hydrogen~ted starch hydrolysate or the like, and (2) a relatively light textured frappe, generally prepared from egg albumin, gelatin, vegetable proteins, such as soy derived cu"l~oullds, sugarless milk derived compounds such as milk proteins, and Illix~ures thereof. The frappe is generally relatively light, and may, for example, range in density from about 0.5 to about 0.7 grams/cc.

The high boiling syrup, or "bob syrup" of the soft confectionery is relatively viscous and has a higher density than the frappe co~ ent, and frequently conf~in~ a substantial amount of carbohydrate bulking agent such as a hydrogenated starch hydrolysate Conventionally, the final nougat composition is pre~ d by the~d~lihon of the "bob syrup" to the frappe under ~git~iQn, to form the basic nougat ll~Lure~ Further ingredients such as flavoring agents, ~ldition~l carbohydrate bulking agent, coloring agents, preservatives, m~liç~ ixLulc;s thereof and the like may be added thereafter also under ~git~tion A general discussion of the composition and a~ion of nougat confections may be found in B.W. Minifie, Chocolate, Cocoa and ConfectioneTy: Science and Technolo~, 2nd edition, AVI Publishing Co., Inc., Westport, Conn. (1980), at pages 424-425, which disclosure is incorporated herein by reference.

The procedure for preparing the soft collr~cLionery involves known procedures. In general, the frappe component is pl~ed first and thereafter the syrup co,l,~ ent is slowly added under agitation at a l~ Luie of at least about 65~C., and preferably at least about 100~C. The l~ixLul~e of components is contin~led to be mixed to form a ullirolll~ ixLule, after which the l~ixLul~i is cooled to a Lc;l~ Lule below 80~C., at which point, the flavoring agent may be added. The llfi~lule is further mi-xed W O96/14868 PCTnUS95/12853 for an additional period until it is ready to be ~ uved and formed into suitablec~nre~;l;onçry shapes.

The ingestible therapeutic wound healing compositions may also be in the form of a ph~rm~cellhc~l suspension. Ph~rm~centi~.~l suspensions of this invention may be y~ Jaled by CO~ 1 ;on~l mrthorlc long established in the art of ph~rm~celltir~l c~ o~ ling SuspenQ~ne may contain adjunct m~ttori~le employed in formlll~ling the p~n~ione of the art. The suspensions of the present invention can comrriee (a) preservatives such as butylated hydroxyanisole (BHA), butylated llydlo~ytoluene (BHT), benzoic acid, ascorbic acid, methyl paraben, propyl paraben, tocopherols, and the like, and llli~l,ll'tS thereof. Pres~.valives are generally present in ~IllUUlltS Up to about 1%, and preferably from about 0.05% to about 0.5%, by weight of the suspension;
(b) buffers such as citric acid-sodium citrate, phosphoric acid-sodium phosphate, and acetic acid-sodium acetate in ~ oullLs Up to about 1%, and preferably from about 0.05% to about 0.5%, by weight of the suspension;
(c) suspending agents or thickeners such as cellulosics like methylcellulose, carrageenans like alginic acid and its derivatives"r~n~h~n gums, gelatin, ~c~c;~e, and microcrystalline ce~ lose in amounts up to about 20%, and preferably from about 1% to about 15%, by weight of the susprneion;
(d) antifoaming agents such as dimethyl polysiloxane in ~IlllUllntS Up to about 0 2%, and preferably from about 0.01% to about 0.1%, by weight of the suspenslon;
(e) ~we~lr~.;..g agents such as those ~w~ rs well known in the art, incll~(ling both natural and artificial sweetton~ne Swe~ lg agents such as monos~crh~ri~les, rlie~r,rh~ les and polys~cch~ri~les such as xylose, ribose, glucose (dextrose), m~nnose~ galactose, fructose (levulose), sucrose (sugar), m~1tose, invert sugar (a l~Lule of fructose and glucose derived from sucrose), partially hydrolyzed starch, corn syrup solids, dihydrochalcones, monellin, steviosides, glycyl.lli;chl, and sugar alcohols such as sorbitol, ~ n~ maltitol, hydrogenated starch hydrolysatesand ~ Lules thereof may be utilized in allluullLs up to about 60%, and preferably from about 20% to about 50%, by weight of the suspension. Water-soluble artificial swe~;Lellers such as soluble saccharin salts, i.e., sodium or calcium s~cch~rin salts, W O96/14868 PCTrUS95/12853 cycla~ salts, the so~ lm, h.~ nlilll~l or c~lcillm salt of 3,4-dihydro-6-methyl-1,2,3-nY~ ,;.le-4-one-2,2~i~xi~e, the pot~ccillm salt of 3,4-dihydro-6-methyl-1,2,3-oY ~Ih;; ~;.-e-4-one-2,2-dioxide (~ceslllf~me-K), the free acid form of .c~crh~rinJ and the like may be utilized in amounts from about 0.001% to about 5%, by weight of the suspension;
(f) flavoring agents such as those flavors well known to the skilled artisan, such as natural and artificial flavors and mints, such as peppc~ , menthol, citrus flavors such as orange and lemon, artificial vanilla, cinn~mnn, various fruit flavors, both individual and mixed and the like may be utilized in amou~lLs from about 0.5% to about 5%, by weight of the sllcpension;
(g) coloring agents such as pigmentc which may be incorporated in ~ulll~ up to about 6%, by weight of the suspension. A p~ertilled pigment, 1;l;.ll;lllll xi~k?, may be incorporated in ~Ulluul-L~ Up to about 2%, and preferably less than about 1%, by weight of the suspension. The coloring agents may also include natural food colors and dyes suitable for food, drug and cosmP~tic applications. These colorants are known as F.D.& C. dyes and lakes. The m~t~ri~l.e acceptable for the foregoing uses are preferably water-soluble. Such dyes are generally present in ~ ullLS up to about 0.25%, and preferably from about 0.05% to about 0.2%, by weight of the suspension;
(h) decolorizing agents such as sodium metabisulfite, ascorbic acid and the like may be incorporated into the suspension to p~evelll color rh~nge~e due to aging.
In general, decolorizing agents may be used in ~ll~LulLS up to about 0.25%, and preferably from about 0.05% to about 0.2%, by weight of the suspension; and (i) solubilizers such as alcohol, propylene glycol, polyethylene glycol, ~-5 and the like may be used to solubilize the n~v~ling agents. In general, solubilizing agents may be used in all~oullL~ Up to about 10%, and preferably from about 2% to about 5%, by weight of the suspension.

The ph~rm~ceutical suspensions of the present invention may be ~0 plv~d as follows:
(A) admix the thickener with water heated from about 40~C. to about 95~C., preferably from about 40~C. to about 70~C., to form a dispersion if the thickener is not water soluble or a solution if the thiçkenPr is water soluble;

W O96/14868 . PCTnUS95112853 (B) admix the ~wc;e~-.;..g agent with water to form a svl~ltiQn;
(C) admix the thel~a~ lic wound healing composition with the thirl~nPr-water ~ ";~ -e to form a ~ irvllll thi~ ner-therapeutic wound healing comrosittQn;
S ~D) combine the ~wc;~ er sol~lti~n with the thirlr~n~.r-therapeutic wound healing composition and mix until ~ ;rv...~, and (E) admix the optional adjunct m~t~ri~l~ such as coloring agents, navv~ g agents, decolorants, solubilizers, ~nbfio~ming agents, buffers and ~ ittt)T
water with the ~ ule of step (D) to form the suspension.
The ingestible therapeutic wound healing compositions of this invention may also be in chewable form. To achieve acceptable stability and quality as well as good taste and mouth feel in a chewable formlll~tic)n several considerations areimportant. These conci(lerations include the amount of active substance per tablet, the flavoring agent employed, the degree of ccl~ s~ibility of the tablet and the organoleptic properties of the composition.

Chewable therapeutic candy is ~l~ared by procedures similar to those used to make soft confectionery. In a typical procedure, a boiled sugar-corn syrup blend is formed to which is added a frappe mixture. The boiled sugar-corn syrup blend may be prepared from sugar and corn syrup blended in parts by weight ratio of about 90:10 to about 10:90. The sugar-corn syrup blend is heated to temp~ L"cs above about 120~C. to remove water and to form a molten mass. The frappe is generally prepared from gelatin, egg albumin, milk proteins such as casein, and vegetable proteins such as soy protein, and the like, which is added to a gelatin solution and rapidly mixed at ambient temperature to form an aerated sponge like mass. The frappe is then added to the molten candy mass and mixed until homogeneous at tempel~ ,sb~ l about 6S~C. and about 120~C.

The ingestible therapeutic wound healing composition of the instant invention can then be added to the homogeneous llli~ as the temperature is lowered to about 65~ C.-95~ C. whereupon ~ ition~l ingredients can then be added such as W O 96/14868 PCTrUS95/12853 navu~ g agents and coloring agents. The form~ t on is further cooled and formed into pieces ûf desired tlim~n~ionc A general ~liccllcsion of the lozenge and chewable tablet forms of S col,fe-il;on--ry may be found in H.A. Li~ and L. T ~r.hm~n) Ph~ centic~l Dosa~e Forms: Tablets Volume 1, Marcel Dekker, Inc., New York, N.Y. at pages 289to 466, which disclosure is incoll,v,aled herein by reference.

In accordance with this invention, therapeutically effective alll~Ulll~ of the therapeutic wound healing compositions of the present invention may be ~11mi~
into the hard and soft confect onp~ry products. These allloull~ are readily ~let~rmin~l by those skilled in the art without the need for undue eA~le' ;111~ ;on. In a ~lcrtil~cd embo-lim~nt, the ingestible therapeutic wound healing composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 10%
and an ingestible vehicle, that is a ph~rm~ce~tic~lly acceptable carrier, in a quantity sufficient to bring the total amount of composition to 100%, by weight the in~est1hle thcl~c.llic wound healing composition. In a more plerell~ emb~limPnt, the ingestible composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 5%, and in a most plcrel~,d embo-lim~-nt, the ingestible composition will comprise the therapeutic wound healing composition in an amount from about 0.1% to about 2%, and an ingestible vehicle in a quantity sufficient to bring the total amount of composition to 100%, by weight the ingestible therapeutic wound healing composition.

The present invention extends to methods of making the ingestible therapeutic wound healing compositions. In such mt-thorls, an ingestible therapeutic wound healing composition is pl~alcd by ~lmi~ing a therà~c.~lically ~ Live amount of the therapeutic wound healing composition with a ph~rrn~seuhcally-acceptable carrier. The a~pal~lus useful in accordance with the present invention comprisesmiAing and heating apparatus well known in the confectionery arts, and therefore the selection of the specific a~pal~alus will be a~a~ to the artisan. The final ingestible therapeutic wound healing composihQnc are readily prepared using metho-lc generally known in the confectionery arts.

W 096/14868 P~llu~55ll2853 The therapeutic wound healing compositions may also be incorporated into chewing gums. In this form of the invention, the chewing gum composition cQnt~in~ a gum base, a bulking agent, the il~vcnlive therapeutic wound healing composition, and various additives.

The gum base employed will vary gready depending upon various factors such as the type of base desired, the conaiatency of gum desired and the other components used in the composition to make the final chewing gum product. The gum base may be any water-insoluble gum base known in the art, and incl~lclç,e those gum bases utilized for chewing gums and bubble gums. Tlluatr~t1ve examples of suitable polymers in gum bases include both natural and synthetic el~a-tc-m~rs and rubbers. For example, those polymers which are suitable as gum bases include, without limit~tion, substances of vegetable origin such as chicle, crown gum, nispero, r~!s~.linh~ jeh~tong~
perillo, niger gutta, tunu, balata, gutta-percha, lechi-capsi, sorva, gutta kay, l~ cs thereof and the like. Synthetic el~atnm~rs such as butadiene-styrene copolymers,polyisobutylene, isobutylene-isoprene copolymers, polyethylene, u~ixlules dhereof and the like are particularly useful.

The gum base may include a non-toxic vinyl polymer, such as polyvinyl acetate and its partial hydrolysate, polyvinyl alcohol, and llli~UlCS thereof. When t li~e~1, the molecular weight of the vinyl polymer may range from about 2,000 up to and including about 94,000.

The amount of gum base employed will vary greatly depending upon various factors such as the type of base used, the con~i~t~-ncy of the gum desired and the other co~,~olle.,~ used in the composition to make the final chewing gum product.
In general, the gum base will be present in ~~ from about 5% to about 94%, by weight of the final chewing gum composition, and preferably in a."ou~ from about15% to about 45%, and more preferably in al~loull~ from about 15% to about 35%, and most preferably in amounts from about 20% to about 30%, by weight of the final chewing gum composition.

The gum base composition may contain cO.lv .~ n~l Çl~etC-m~r solvents to aid in sort~ g the el~et~m~r base co~onent. Such Pl~et m~r solvents _ay ccmpri.ee terpinene resins such as polymers of Alpha-pinene or ~-pinene, methyl,glycerol or pentaerythritol esters of rosins or motlified rosins and gums, such as hydrogenated, ~lim~ri~d or poly...k.;~ed rosins or l~ules thereof. Examples of etC~m~r solvents suitable for use herein include the pentaery~ritol ester of partially hydrogenated wood or gum rosin, the pentaelyl~ ol ester of wood or gum rosin, the glycerol ester of wood rosin, the glycerol ester of partially ~1;".. . ;~ed wood or gum rosin, the glycerol ester of polym~ri7ed wood or gum rosin, the glycerol ester of tall oil rosin, the glycerol ester of wood or gum rosin and the partially hydrogenated wood or gum rosin and the partially hydrogen~ted methyl ester of wood or rosin, llliALulGs ~ereof, and the like. The e1~etomer solvent may be employed in amounts from about 5% to about 75%, by weight of the gum base, and preferably from about 45% to about 70%, by weight of the gum base.
A variety of traditlQn~l ingredients may be included in the gum base in G~GCLiVe alllOUlltS such as pl~etici7~rs or softeners such as lanolin, p~lmilic acid, oleic acid, stearic acid, sodium ste~r~te potassium ste~r~te7 glyceryl tri~cet~te7 glyceryl ecithin, glyceryl monoste~rate propylene glycol m-)noste~rate acetylated monoglyceride, glycerine, miAlU~GS thereof, and the like may also be incorporated into the gum base to obtain a variety of desirable l~Alul~s and c~n~ictency properties.
Waxes, for example, natural and synthetic waxes, hydrogenated vegetable oils, petroleum waxes such as polyurethane waAes, polyethylene waxes, p~r~ffin waxes, microcrystalline waxes, fatty waxes, so,l,i~ monoste~rate, tallow, propylene glycol, ~AlUIeS thereof, and the like may also be incGl~olal~d into the gum base to obtain a variety of desirable l~;~lu-es and c~n~ietency properties. These tra~lit1c)n~l a(ltlit10n~1 m~teri~1.e are generally employed in amounts up to about 30%, by weight of the gum base, and preferably in allWUlll.~7 from about 3% to about 20%, by weight of the gum base.
The gum base may include effective ~llOUlll:i of mineral adjuv~l~ such as calcium carbonate, m~gnç~eillm carbonate, all min~ .";.l~l,ll hydroxide, s311llll;lllllll silicate, talc, tricalcium phosphate, ~lic~lcillm phosph~te and the like as well as WO 96/14868 PCTtUS95/12853 ~LIUi~S thereof. These mineral adjuvallls may serve as fillers and textural agents.
These fillers or alju~ ls may be used in the gum base in various ~..o~ Preferably the amount of filler when used will be present in an amount up to about 60%, by weight of the chewing gum base.

The chewing gum base may ac~ tion~lly include the cc,~ .,l;rn~l additives of coloring agents, ~ntioxi-l~ntc, plest.vàLives and the like. For example, -,;,,.,, dioxide and other dyes suitable for food, dIug and cQsmetiC applir~1ionc, known as F.D. & C. dyes, may be l1tili7eA An ~ntioxi-l~nt such as butylated 10hydru~ytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, and ~luies thereof, may also be inclllde~l Other c~ vG~l;cn~l chewing gum additives known to one having ordinary skill in the chewing gum art may also be used in the chewing gum base.

15The gum composition may include effective ~ ; of collvt~l-L;Qn~l additives selected from the group con~icting of sweetening agents (sweeteners), pl~chci7t nc solL~I~e. j, emlllcifiP,rs, waxes, fillers, bulking agents, mineral adjuvallts, navol;llg agents (flavors, n~v~ gs), coloring agents (colorants, coloringc)~
~nti~xitl~ntc, ~cicl--l~ntc, thir~ nP~rS~ lU1GS thereof and the like. Some of these additives may serve more than one purpose. For example, in s.. g~rless gum col,l~osilions, the sweetrner~ e.g., sorbitol or other sugar alcohol or llfL~lulGs thereof, may also function as a bulking agent. Simil~rly, in sugar conl;.;.,;.,g gum compositions, the sugar ~ww~c~ can also function âS a bulking agent.

25The plastici_ers, softeners, mineral adjuv~lL~, colorants, waxes and ~n~io~ ntc ~liccllc~secl above as being suitable for use in the gum base may also be used in the gum composition. Fx~mples of other COIIV~ ,I;rm~l additives which may be used include emlllcifiers~ such as lecithin and glyceryl monoste~r~te~ thirl~nerc, used alone or in combination with other softrners~ such as me~yl cellulose, ~l in~tes, 30carrageenan, ~nth~n gum, gelatin, carob, tr~g~c~nth, locust bean, and carboxy methyl cellulose, ~ci~lnl~ntc such as malic acid, adipic acid, citric acid, tartaTic acid, fumaIic acid, and llli~lUlGs thereof, and fillers, such as those ~liccucsecl above under the WO 96/14868 PCT/US95tl2853 calcgu.y of mineral adjuv~lls. The fillers when used may be utilized in an ~lluullt up to about 60%, by weight of the gum colllposilion.

Bulking agents (carriers, e~rten-l~rs) suitable for use in chewing gums include ~w~e~ g agents srlP!ct~l from the group con~ ting of m-)nos~crh~ri<les, cr.h~rides, poly-s~crh~n~les, sugar alcohols, and IlliAlul~s thereof; polydcAllose;
_altode"ctnn.c; minerals, such as c~lcinm carbonate, talc, ~ 111 clioxi-le, dic~lcillm phosph~te and the like. Bulking agents may be used in a~ unls up to about 90%, by weight of the final gum composition, with ~IllUWlL:~ from about 40% to about 70%, by weight of the gum composition being ~lc~cllcd, with from about 50% to about 65%,by weight, being more ple~ ,d and from about 55% to about 60%, by weight of the chewing gum composition, being most plcîcllcd.

The sweetening agent used may be selected from a wide range of m~teri~ inchl~ling water-soluble sweeteners, water-soluble artificial sweeteners, water-soluble sweeteners derived from naturally oCcllrring water-soluble sweeteners, dipeptide based ~wcel~ners, and protein based sweetenrrs, including llfiAlules thereof. Without being limited to particular sweeteners, reprcselltillive c~tegoriee and examples incl~l(le:
(a) water-soluble ~wc~ g agents such as monos~cr. h~rides, ~ cch~rides and polysaccharides such as xylose, ribulose, glucose (dextrose), m~nnose galactose, fructose (levulose), sucrose (sugar), m~ltose, invert sugar (a ~ AtUi'~ of fructose and glucose derived from sucrose), partially hydrolyzed starch, corn syrup solids, dihydrochalcones, m-)nellin, steviosides, glycyrrhi_in, and sugar alcohols such as sorbitol, Ill~ 1, maltitol, hydrogenated starch hydrolysates and ll.,~lwcs thereof;
(b) water-soluble artificial ~Wc~ rr~s such as soluble s~crh~rin salts, i.e., sodium or calcium saccharin salts, ~iycla~ c salts, the so~ lm, ~ or calciumsaltof3,4-dihydro-6-methyl-1,2,3-ox~thi~7.ine-4-one-2,2-dioxide, thepot~.~sillm salt of 3,4-dihydro-6-methyl-1,2,3-ox~thi~7ine-4-one-2,2-dioxide (~cesnlf~me-K), the free acid form of saccharin, and the like;
(c) dipeptide based ~we~t~.çrs~ such as L-aspartic acid derived sweeteners, such as L-aspartyl-L-phenyl~l~nine methyl ester (Aspartame) and m~teri~l~
described in United States Patent No. 3,492,131, ~Alpha-aspartyl-N-(2,2,4,4-W O96/14868 PCTrUSgS/12853 yl-3-thietanyl)-D-alanin-amide hydrate(Alitame),methylestersofL-aspartyl-~phenylglycerineandL-aspartyl-L-2,S-dil-yLophenyl-glycine, L-aspartyl-2,5-dihydro-L,phenylalanine; L-aspartyl-L-(1-cycl- heYen)-al~nine and the like;
(d) water-soluble sweeteners derived from naturally occurring water-Ssoluble ~w~ n~r~ such as ~.hlorin~t~l dc.;v~ives of ordinary sugar (sucrose), known, for example, under the product deeigr ~tion of Sucr~lose; and (e) protein based ~we~ .er~ such as th~nm~nccous danielli ~h~.. l;.
I and r[).

In general, an effective amount of sw~ .. er is utilized to provide the level of bulk and/or sweetness desired, and this amount will vary with the sweetener selected This amount of sweetener will non~lly be present in ~uun~ from about 0.002S% to about 90%, by weight of the gum cu~ osilion, depending upon the sweetener used. The exact range of a~nuun~ for each type of sweetener is well known lS in the art and is not the subject of the present invention. The amount of sweetener ordinarily necessary to achieve the desired level of sweetness is independent from the flavor level achieved from flavor oils.

Pl~r~lled sugar based-sweeteners are sugar (sucrose), corn syrup and ~ ulcS thereof. Preferred sugarless sweeteners are the sugar alcohols, artificial sweeteners, dipeptide based ~w~lG..ers and "i~ s thereof. Preferably, sugar alcohols are used in the sugarless compositions because these sweet~ner~ can be used in ~l(~LIn~ which are sufficient to provide bulk as well as the desired level ofsweet~e~s ~,rti~cd sugar alcohols are selçcted from the group con~ hng of sorbitol, xylitol, maltitol, .. ~ ol, and~ Lules thereof. Morepreferably, sorbitol oral~
of sorbitol and ".~ .1 is llhli7ed The gamma form of sorbitol is ~ ;Çt;"Gd. An artificial sweetener or dipeptide based sweetener is preferably added to the gumcompositions which contain sugar alcohol~

The coloring agents useful in the gum compositions are used in a~,uu~
effective to produce the desired color. These coloring agents include pi~n~nti which may be incorporated in ~llOUI~ up to about 6% by weight of the gum composition.
A ~Ic;r~,l-ed pigment, ~ i()xi(le, may be incorporated in ~lluun~ up to about W O96/14868 PCTrUS95/12853 2%, and preferably less than about 1% by weight of the composition. The colorants m ay also include na~hural food colors and dyes suitable for food, drug and cosmtoh!c applir,?~h~n~ These colorants are known as F.D.& C. dyes and lakes. The m~teri~l~
~ acceptable for the foregoing uses are preferably water-soluble. Illustrative nonlimitin~
examples include the indigoid dye known as F.D.& C. Blue No.2, which is the o~1ium salt of 5,5-indigohn~ .llfonic acid. gimil~rly, the dye known as F.D.~ C.Green No.1 comprises a hiphenylm~th~ne dye and is the m~noso~ m salt of 4-[4-~N-ethyl-p-sul~niullibenzylamino~liph~ ylllwlllylene]-[1-(N-ethyl-N-p-s~.l r~...;.,."benzyl)-delta-2,5-cyclohexadieneimine]. A full recitation of all F.D.& C. colorants and their coll~,~onding ch~mic~l structures may be found in the Kirk-Othmer Enc.,rclopedia of Chemical Technolo~, 3rd Fclihon, in volume 5 at pages 857-884, which text is incorporated herein by reference.

Suitable oils and fats usable in gum compositions include partially hydrogenated vegetable or animal fats, such as coconut oil, palm kernel oil, beef tallow, lard, and the like. These ingredients when used are generally present ina-.loull~ up to about 7%, by weight, and preferably up to about 3.5%, by weight of the gum composition.

In accordance with this invention, therapeutically effective amounts of the therapeutic wound healing compositions of the present invention may be ~-imixed into a chewing gum. These a~ are readily ~letermined by those skilled in the artwithout the need for undue expprim~nt~hon In a prerelled embo-lim~nt, the final chewing gum composition will compri~e the therapeutic wound healing composition in an amount from about 0.1% to about 10% and a chewing gum composition in a quantity sllffirient to bring the total ~ml-nnt of composition to 100%, by weight of the chewing gum composition. In a more pl~rwl~id embo~iimrnt~ the final chewing gum composition will comprise the therapeutic wound healing composition in an amountfrom about 0.1% to about 5%, and in a most prc~ d emboflimt-nt, the final chewing gum composition will compri.~e the thwrapeutic wound healing composition in an ~mmlnt from about 0.1% to about 2%, and a chewing gum composition in a quantity sllffiriPnt to bring the total amount of composition to 100%, by weight of the chewing gum composition.

The present invention extends to methods of making the therapeutic chewing gum compositions. The the ~ ;c wound healing compoeitic-ni may be incorporated into an otherwise collv~ >n~l chewing gum composition using standard techniques and eq~lirm~nt known to those skilled in the art. The apparatus useful in accor~ce with the present invention collllll;eçs mixing and heating a~a-~us wellknown in the chewing gum m~nllf~ctllring arts, and therefore the selecti~n of the specific a~ lus will be apparent to the artisan.

For example, a gum base is heated to a L~ eLdluic; snfficiently high enough to soften the base without adversely effecting the physical and chemical make up of the base. The Oplilllulll temperatures utilized may vary depending upon the composition of the gum base used, but such tellll,cr~ules are readily ~1ett-rminerl by those skilled in the art without undue c~l~c ;~ ;on The gum base is conventionally melted at ~tlll~)C~UIeS that range from about 60~ ~. to about 120~ C. for a period of time sufficient to render the base molten.
For example, the gum base may be heated under these con~1ihone for a period of about thirty lllillUl~S just prior to being a-lmixed incl~ 11y with the le~ ;ll;llg ingredients of the base such as the pl~etici7t-r, fillers, the bulking agent andlor sweet~ner~, the softener and coloring agents to pl~etici7e the blend as well as to modlll~te the hardness, viscoelasticity and formability of the base. The chewing gum base is then blended with the therapeutic wound healing composition of the present invention which may have been previously blended with other tr~lition~l ingredients. Mixing is continued until a ullir~ ul~ of gum composition is obtained. The~rl~l the gum composition I~ u,~; may be formed into desirable chewing gum shapes.

In a specific embolimt-nt, the invention is di~cL~d to a therapeutic ph~rms~celltic~l composition for ~r~vel~ g and red~lcing injury to ",i.."",~ n cells, and increasing the resuscitation rate of injured ~ n cells, which c~mrrieçs (A) a therapeutically effective amount of a therapeutic wound healing composition of Embodiment One (I) selecte(l from the group c~neiehng of:

a.A) (a) pyruvate selecte(l from the group concicting of p-yruvic acid, ph~rm~celltic.~lly acceptable salt~c of pyruvic acid, and mi~lwes thereof;
(h) an ~nti~xi-l~nt; and (c) a ~ we of s~hlr~te(l and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular ,.. ~ C and rÇsllcçit~tion of l,.. "~ n cells;

a.B) (a) ~yluv~l~ selecte~l from the group cQn~ ng of p-yruvic acid, ph~rm~cellhcally acceptable salts of pyruvic acid, and llli~Lwes thereof;
(b) lactate s~lecte~l from the group conci~ting of lactic acid, ph~rm~celltically acceptable salts of lactic acid, and l~Lw~s thereof; and (c) a ~ Lw~; of .c~tllr~tetl and nn.c~tllr~ted fatty acids wherein the fatty acids are those fatty acids required for the repa* of cellular m~.rnhr~nes and resuscitation of ",~ "n~ n cells;
(I.C) (a) an antioxidant; and (h) a l~ Lw~i of saturated and unsaLw~ted fatty acids wherein the fatty acids are those fatty acids requ*ed for the repair of cellular lllcl~~ es and res~lccit~tion of n-~ n çells;
(I.D) (a) lactate select~d from the group concicting of lactic acid, ph~rm~celltic.~lly acceptable salts of lactic acid, and ll~Lw~es thereof;
(b) an antioxidant; and (c) a ~i~lwe of s~tllr~tecl and nns~tllr~tecl fatty acids wherein the fatty acids are those fatty acids requ*ed for the repa* of cellular mr.mhr~nes and resllcr.it~hQn of ".~ n cells; and (B) a ph~rm~r,ellhc~lly ~ccept~ble carrier.

The ph~rm~cel_tically acceptable carrier may be selecte(l from the group cQnci.chng of ph~rm~cel1hc.~1 appliances, topical vehicles, and ingestible vehicle.

In another specific embo-lim~.nt, the invention is directed to a method for p~ lg a therapeutic ph~rm~celltical composition for preventing and redncing WO 96/14868 . PCT/US95/12853 injury to ~ "~ n cells, and increasing the resllccitAhon rate of injured ~ ..llll,Ali~n cells, which comprises the steps of:
(A) providing a therape~lhc~lly ~iliv~i amount of a therapeutic wound healing composition of Embodiment One (I.A-D) selecte(l from the group concictin~ of:
s (I.A) (a) ~yl~lva~t; selected from the group c-~n~i~hng of pyruvic acid, ph~rmAcellhcAlly acceptable salts of pyruvic acid, and ~lf~ es thereof;
(b) an antioxidant; and (c) a llfi~ule of saturated and uncAhlr~ted fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and reclleçit~hsn of, ~ AliAn cells;

(I.B) (a) pyruvate selected from the group con~icting of pyruvic acid, ph~rm~cel~tieAlly acceptable salts of pyruvic acid, and llfi~Lulc;s thereof;
(b) lactate selected from the group co~cichng of lactic acid, ph~rmAcel_tically acceptable salts of lactic acid, and ~ S thereof; and (c) a ll~i~ e of s~lrAted and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repa* of cellular membranes and resuscitation of l,~ -.. Ali~n cells;
(I.C) (a) an ~ntis~ nt; and (b) a ll~Lule of sa~ ted and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of ~ llllllAliAn cells;
(LD) (a) lactate s~-,lected from the group concicting of lactic acid, ph~rmAceutically acceptable salts of lactic acid, and ~UI~S thereof;
(b) an antioxidant; and (c) a l~i~Lule of saturated and unsaturated fatty acids where*n the fatty acids are those fatty acids required for the repair of cellular lllelll~l~les and r~sncc.itAtisn of ",~""~Ali~n cells; and (B) providing a ph~ cellhc~lly acceptable carrier; and (C) ~1mixing the therapeutic wound healing composition from step (A) and the ph~rm~cel-ti~lly acceptable carrier from step ~B) to form a therapeutic ph~rm~cenhc~
composition.

S Throughout this applic~tic-n, various publication~ have been referenced.
The fli~closllres in these publications are inco,~vlaled herein by reference in order to more fully describe the state of the art.

The present invention is further illustrated by the following examples which are not inten~l to limit the t;rreclive scope of the claims. All parts andpercentages in the examples and throughout the specification and clairns are by weight of the final composition unless otherwise specified.

E. Examples Of The Therapeutic Wound Healing Compositions Of Embodiment One (I.A-D) Study 1 This study dt-m-)n~trates a comr~ri~on of the viability of U937 monocytic cells after exposure of the cells to various ~ntioxid~nt~ and combin~t on~
of antioxidants. This study also demonstrate a cu...~ oll of the levels of hydrogen peroxide produced by U937 monocytic cells and ",i1."".~ n epi-l~rm~l k~r~tinocytes after exposure of the cells to various ~ntioxi(3~nt~ and combin~t on~ of ~ntioxirl~nt~
The results of this study are ilh~tr~ted in Figures 1~ and examples 1-26 below.

M~mm~ n epidermal k~tinocytes and monocytes were employed to eY~mine the ability of various antioxidants to reduce levels of hydrogen peroxide in these cells. Hydrogen peroxide was measured after the cells were exposed to ultraviolet light in the wavelength range from 290 to 320 nm (UV-B) or to the infl~..",.~ y compound 12-0-tetr~clec~noyl-phorbol-13-acetate (TPA). Sodium pyruvate was tested at various concpntr~ion~ to determine the effect of concentrations of this antioxidant on the hydrogen peroxide production by epidermal cells and CA 02204l4S l997-04-30 W O96/14868 PCTrUS95/12853 monocytes. M~gnç~illm pyruvate, c~lcillm pyluvalt;, zinc ~yluv~Le, and cc~ ;on~
of sodium ~yluvale with ascorbic acid, lactic acid, and Vitarnin E were then tested to e the effect of these salts and cc,~l.;.~;on.~ of ~nt1oxitl~nt.~ on the hydrogenperoxide production by epidermal cells and monocytes.
M~mm~ n epidermal k~tinocytes were isolated by tryp~ini~tion of epithelial sheets and grown in mol1ifit~1 basal MCDB 153 m~lillm suppl~ (1 with epidermal growth factor, bovine ~iluiL~y extract, and hydrocortisone. Cells were;-led in a hl]mi(lified incubator with 5% carbon dioxide at 37~C. Keratinocytes were seeded in 60 rnm culture dishes at a cell density of 3 x 105 cells per dish and the cultures were exposed to 1 M.E.D. dose of ultraviolet-B light (100 mJ/cm2) or treated with 100 ng/ml of TPA.

U937 monocy~ic cells are a cultured cell line grown in RPMI media with 10% fetal calf serum. Cells were .. ~ ;necl in a 60 mm culture dish at 5% carbondioxide at 37~C. at a see~1ing density not ey~eeflin~ 1 x 106 cells per dish.

Sodium pyluva~e, lactic acid, ascorbic acid, and Vitamin E were dissolved in ~ tille(l water, with s~lffici~nt surf~ct~nt The concentrations of the sodium pyruvate solutions prepared were 1 rnM, 10 mM, 50 mM, 100 rnM, and 200 mM. The concentrations of the lactic acid solutions prepared were 1.0%, 0.1%, and 0.05%. The concentrations of the ascorbic acid solutions p~ aLcd were 1.0%, 0.1%, 0.05%, and 0.025%. The concPntr~t ltn~ of the Vitamin E solutions ~l~aL~
were 1 U, 10 U, 50 U, and 100 U. The test solutions were adjusted to a pH value of 7.4 with 1.0N sodium hy~ide solution and then sterile filtered. The a~l~iale conct~ntration of test solution or combination of test solutions was added to the cells imm~i~tely prior to exposure of the cells to ultraviolet light-B or TPA [lOOng/ml].
Stock solutions were ~rcL)~cd so that the vehicle did not con~tihlte more than 1% of the total volume of the culture media.
Intr~ctollnl~r hydrogen peroxide prol~1ction by ~ .."..~ n eFi(l~m~l keratinocytes and U937 monocytes was measured using dichlorofluorescein ~ cet~t~(DCFH-DA, Molecular Probes, Eugene, Ore.). DCFH-DA is a non-polar non-W O96/14868 PCTrUS95/12853 fluolesc~ com~ ulld that readily ~ es into cells where it is hydrolyzed to the polar non-nu~ sc~ll d~;valive DCFH which then beco.l,es trapped within the cells. In the presence of intr~ce~ r hydrogen peroxide, DCFH is c-xi~ ed to the highly fluolescel.l - compound DCF. Hence, cellular fluorescence intensity is directly ~r~lional to the S level of intr~celllll~r hydrogen peroxide pro hl(cP~d Cellular fluorescPnce intensity can be l~o~ oled by fll~imt?try and by flow c~

~r~mm~ n epidermal keratinocytes and U937 cultured monocytes (I
x 106 per dish) were incnb~tP~l at 37~C. with S uM of DCFH-DA. Production of hydrogen peroxide was measured using a Coulter Profile analytical flow c~ lc;tel.
Linear and log intensity of green fluoresc~nce data was collected. For each analysis, a quantity of 10,000 to 20,000 events was accum.ll~te~l Optical ~lignm~nt for the in~ ,e..l was p~-ro-lllcd daily. Coefficie~t~ of variation for rulwal.l angle light scatter and it~ ,r~le-l green fluorescence were generally less than two. Each analysis was repeated three times and the 4~ ;Qn of fluorescence was cA~Iessed in terms of r~ O~ les (fmol, 10-15 moles) of DCF o~ i7~l per cell, which is a direct measure of the intracell~ r hydrogen peroxide produced. Altematively, in the saturated and unsaturated fatty acid examples in examples 27-52, fluoli~llelly was used to assess the DCF oxidation per cell.
The viability of the U937 monocytic cells after eYrosllre of the cells to various antioxidants for 24 hours was measured. The viability of the cells was determine(l by exposing the cells to the dye propidium iodide. Permeable cell me~ es which absorbed the dye were not considered viable. The viability of the cells was ~~,~-Gs~ t;d as the pe.~;t;ll~ge of cells that eYchl~3ed propidium iodide.
Figure 1 depicts in bar graph format the viability of U937 monocytic cells afterexposure of the cells to no ~ntioxi-l~nt (Example 1, control), to sodium pyruvate (Example 2), to ascorbic acid (Example 3), to lactic acid (Example 4), and to Vitamin E (Example 5). Figure 2 depicts in bar graph format the viability of U937 monocytic cells a~er exposure of the cells to various combin~hQn~ of ~nh~xirl~nt~
Specifically, the viability of U937 monocytic cells was measured after exposure to no ~nh~xid?nt (Example 6, control), to ascorbic acid and lactic acid (Exarnple 7), to ascorbic acid and Vitamin E (Example 8), to sodium pyruvate and ascorbic acid -(Example 9), to sodium pyluva~e and lactic acid (Example 10), to sodium l~yluv~le and Vitamin E (Example 11), to lactic acid and Vitamin E (Example 12), and to sodium~luv~le, ascorbic acid, and lactic acid (~Y~mrle 13).

Figure 1 shows that ascorbic acid is CylO~iC to monocytes at conç~ntrations as low as 0.25%. Figure 2 shows that the cytotoxicity of ascorbic acid was l~velsed by the addition of 10 mM of sodium pyruvate. Figures 1 and 2 show that the viability rate of 15% to 20% of the cells when treated with ascorbic acid was increased to 95% to 98% upon ~ litinn of sodium pyruvate. Lactic acid and Vitamin E did not reverse the cytotoxicity of ascorbic acid.

Sodium pyruvate was then tested at various concentr~hc n~ to clet~rmine the effect of concentrations of this ~nhoxi(l~nt on the hydrogen peroxide production by epidermal cells and monocytes. M~mm~ n epidermal keratinocytes and monocytes were exposed to (a) 1 M.E.D. dose of ultraviolet light-B and (b) 100 ng/ml of 12-O-tetr~-lec~ncylphorbol-13-acetate (TPA) in the presence of sodium pyruvate at thefollowing concentrations: 200 mM, 100 mM, 50 mM, 10 rnM, 1 mM.

The opLiLuulll concentr~hon of sodium pyluv~t~ to reduce the hydrogen peroxide production by epidermal cells and monocytes was found to be 10 mM.
Concentrations of sodium py.uv~e of 50 rnM and above were ~iy~O~iC to both epidermal keratinocytes and monocytes.

M~gnç~illm pyruvate, c~lcium pyruvate, zinc pyluv~, ascorbic acid, lactic acid, and Vitamin E, and combin~tion~ of sodium pyluvalti with ascorbic acid, lactic acid, and Vitamin E were then tested to ~letennine the effect of these salts and combinations of ~ntioxi~l~nt~ on the hydrogen per~xide production by epidermal cells and monocytes. The following test solutions were p.~alcd.

(a) sodium pyruvate C10 mMl;
(b) zinc salt [10 m~;
(c) m~gnesium salt [10 mM];

W O96/14868 PCTrUS95/12853 (d) calcium salt [10 mMl;
(e) sodiurn pyruvate [10 mMl and ascorbic acid [0.025%];
(f) sodium ~yluv~te [10 mMl and lactic acid [0.05%], (g) sodiurn pyruvate [10 rn~, lactic acid, [O.OS%], and ascorbic acid [0.025%];
(h) lactic acid [1.0%, 0.1%, and 0.05%];
(i) ascorbic acid [1.0%, 0.1%, 0.05%, and 0.025%];
~) Vitamin E [1 U, 10 U, 50 U, and 100 U]; and (k) vehicle solvent controls.

There was no significant difference arnong the zinc, magnesium, and c~lcillm salts of pyruvic acid on the hydrogen peroxide pro l~lct on by epidermal cells and monocytes. The zinc and calcium salts of pyruvic acid intll1ced di~.~ iation of keratinocytes. For co-~v~nience, the sodium salt was used in subsequent tests.

The ~~ lum concentration of lactic acid to reduce the hydrogen peroxide production by epidermal cells and monocytes was found to be 0.05%. The Opli~ concen~ on of ascorbic acid was found to be 0.025%. The higher c~nc~-n1rations of both of these compounds were found to be ~iy~ic to both typesof cells. The Op~lllu~ll concen~nn of Vitamin E was found to be 50 U.

Figure 3 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells after exposure of the cells to no ~n~i-~xi(1~nt(Exarnple 14, control), to sodium pyl.~valt; (Example 15), to ascorbic acid (Example 16), to lactic acid (Example 17), and to Vitamin E (Example 18). Sodiumpyruvate and Vitamin E significantly reduced the hydrogen peroxide production bymonocytes.

Figure 4 depicts in bar graph format the levels of hydrogen peroxide produced by U937 monocytic cells after exposure of the cells to various combin~tion~

W O96/14868 PCTrUS95/12853 of ~nhoxi-l~nt~ Specifically, the levels of hydl~ll peroxide produced by U937 monocy-tic cells were measured after ~ O~ul~ to no ~nhoxi~1~nt (F.Y~mrle 19, control), to ascorbic acid and lactic acid (FY~mrle 20), to ascorbic acid and Vitamin E
~Example 21), to sodium pyruvate and ascorbic acid (Example 22), to sodium py~uvale S and lactic acid (Example 23), to sodium pyluv~l~ and Vitamin E (PY~nlrle 24), to lactic acid and Vitamin E (Example 25), and to sodium pyluv~t~, ascorbic acid, and lactic acid (Example 26). The combination of lactic acid (0.05~/O) and Vitamin E(50 U) si~nific~ntly reduced the hydrogen peroxide production by monocytes.

The morphological alterations in epidermal keratinocytes were observed in control cultures and in cultures eYrosed to ultraviolet-B. Cells in the layer closest to the derrnis are basal ker~hnocytes. These cells proliferate and mi~te into the spinous and ~n~ r layers of the epi(l~rmi~ where the cells begin to dirr~,~.ltiate. The di~.~..t;~hon pattern results in cells en~lcle~hng and fomming cornified envelopes at the uppermost portion of the epidt~.mmi~ the statim comellm The di~.~ ;on of k~r~hnocytes is controlled by the levels of c~lcinm~ m~gne~illm, and other elernPnt~ in the ...~l;...., Cells in culture systems ~lv~ t~g dirr~ ;on appear as an epi-l~rm~l sheet fo~ning ~ hm~ t~ or tight junction~ with each other. K~r~hnocytes that become non~-lh~rent or float in the media were considered responding to a CylOlO~iC
event.

The following morphological ~lt~r~hrmi in the ",A."",~ n epidermal k~hnocytes were observed for the following control cultures:

10 mM Sodium Py~uv~le. Tight junctions of cells were formed and the proliferation rate of the cells was higher than the rate of the control cells.
.

0.025% Ascorbic Acid: Cells were floating in a cytotoxic response to ascorbic acid.

0.025% Ascorbic acid and 10 mM Sodium P~ V~I~; Few tight jnnction~ of cells were observed and cells appeared similar to the cells in the sodium pyluv~te culture.

~ 70 W O96/14868 PCTnUS95/12853 0.05% Lactic Acid: Cells appeared drAmAticAlly altered as an epidermal sheet and as flat ~nulAr cells.

0.05% Lactic Acid and 10 mM So lium p~rrUvate: Cells formed an epidermal sheet but appeared smaller than the cell in the lactic acid culture.

50 U Vitamin E: Cells appeared the same as the cells in the control culture.

50 U Vitamin E and 10 mM Sodium Pyruvate: Cells increased in number and changed in appearance resembling the cells in the sodium pyruvate culture.

The following morphological alterations in the ~A~ AliAn epidermal k~r~tinocytes were observed for the c~ lle~ol~ding cultures exposed to ultraviolet light-B, 100 mJoules, for 24 hours:
10 mM Sodium Pyruvate: Cells proliferated more rapidly than the cells in the control culture.

0.025% Ascorbic Acid: Cells were non~lh~rent and flo~ting in a Cylvt~iC responseto ascorbic acid greater than the cytotoxic response of the corresponding cells without ultraviolet-B light exposure.

0.05% Lactic Acid: Cells formed an epidermal sheet and were more granular than cells in the control culture without ultraviolet-B light exposure.
50 U Vitamin E: Cell growth was inhibited but cells appeared sirnilar to cells in the control culture without ultraviolet-B light exposure.

50 U Vitamin E and 10 mM Sodium Pyluvale: Cells appeared similar to cells in thecontrol culture and proliferated to a greater extent than cells in the control cultures without ultraviolet-B light exposure.

WO 96/14868 -= PCT/US95112853 Morphological ~lterat1l~nc in the U937 monocytic cell line were also obs_,ved for control cultures and cultures eYpose~l to ultraviolet light-B, 100 rnJoules, for 24 hours. The following co~~ ds and combination of compounds, at the concentrations set out below, significantly inhibited the levels of hydrogen peroxide S produced by U937 monocytic cells Sodium pyruvate at 10 mM and 50 mM;
Vitamin E at 50 U and 100 U; and Lactic acid at 0.05% and Vitamin E at 50 U.

E~amples Of The Therapeutic Wound Healing Co~ s 'i~ns of F.mho~liment One (I.A-D) Study 2 This study ~ m~nctratelc a comr~ricon ofthe levels of hydrogen peroxide produced by U937 monocytic cells and epidermal keratinocytes after exposure of the cells to various combin~t on~c of ~ntic)x~ ntc with and without a ll~i~LlllC of s~lul~lcd and nn.c~t~lr~t~d fatty acids. The results of this study are ilhl.ctr~te l in Figures 5-7 and examples 27-52 below.

~ramm~ n epidermal kerattnocytes and U937 monocytic cells and the test solutions of sodium pyruvate, lactic acid, ascorbic acid, and Vitamin E were prepared as describe above for Exarnples 1-26. Tntr~cce~ r hydrogen peroxide pro~hlct-Qn by the "~i"",~ n epidermal keratinocytes and U937 monocytes was alsomeasured as described above.

A llfi~Lu.c of fatty acids derived from chicken fat was pl~ed for ~lition to the cultured cells by mixing 0.1% of the t~.~ic~.n fat with the culture media.
At the temperature of the culture media, 37 ~C., the cllic~en fat was miscible. This chicken fat mixture was added to cultures of cells prior to exposure of the cells to ultraviolet-B light or TPA tre~tm~nt W O 96/14868 . PCTrUS95/12853 As set out in examples 1-26",.~."".~ n epidermal ker~hnocytes and ~nocytes were exposed to (a) 1 M.E.D. dose of ultraviolet light-B and (b) 100 ng/ml of 12-O-tetr~dec~noylphorbol-13-acetate in the presence of various ~nhr-Yi-l~nt~ and combin~h~n~ of ~nhoxill~nt~ with and without a ~lul~; of s~ l~l and lm~
fatty acids [0.1%, 0.5%, and 1.0% cl~ en fat].

Figure 5 depicts in bar graph format the levels of hydrogen pcr~Aide produced by U937 monocytic cells after exposure of the cells to various combin~h-~n.~
of ~nhQXi~l~nt~ with and without a llfi~Lule of saturated and lln~ teA fatty acids.
~pecific~lly~ the levels of hydrogen peroxide produced by U937 monocytic cells were measured after exposure to lactic acid and Vitamin E without fatty acids (Example 27) and with fatty acids (Example 28), to ascorbic acid and lactic acid wi~out fatty acids (Example 29) and with fatty acids (Example 30), and to ascorbic acid and Vitamin E
without fatty acids (Example 31) and with fatty acids (Example 32). The ability of the lS combin~hon~ of lactic acid and Vitamin E, ascorbic acid and lactic acid, and ascorbic acid and Vitamin E to reduce the hydrogen peroxide production by monocytes was increased in the presence of fatty acids. The most ~;~clive combination to reduce the hydrogen peroxide pro lllchon of monocytes was lactic acid (0.05%) and Vitamin E(50 E) in the presence of a ~ e of s~LuraLed and nn.~hlr~t~d fatty acids (0.5%).
Figure 6 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal ker~hnocytes after exposure of the cells to various ~nhoxi-l~nt~
with and without a ~Lule of s~LLulaled and lm~ 7t~1 fatty acids. Spe~ific~lly~ the levels of hydrogen peroxide produced by epidermal k~or~hnocytes were measured after ~o~ to no antioxidant without fatty acids (Example 33, control) and with fatty acids (Example 34), to sodium pyruvate without fatty acids (Example 35) and withfatty acids (Example 36), to ascorbic acid without fatty acids (Example 37) and with fatty acids (Example 38), to lactic acid without fatty acids (Example 39) and with fatty acids (Example 40), and to Vitamin E without fatty acids (Example 41) and with fatty acids (Example 42). The ability of sodium pyruvate and Vitamin E to reduce the hydrogen peroxide production by epidermal ker~hnocytes was increased in the presence of fatty acids. The most effective combin~hon~ to reduce the hydrogen peroxide production of epidermal keratinocytes were sodium pyl~lv~L~ in combination with a WO g6/14868 P~ 55/12853 ~lu~e s~ ed and unsalu-al~d fatty acids and Vitamin E in combination with a Ul~ of s~ andlm~q~ d fatty acids.

Figure 7 depicts in bar graph format the levels of hydrogen peroxide produced by epidermal ke~tinocytes after ~Yposllre of the cells to various col.ll.;.. ~ti- nq of ~nti~xi-l~nt.q with and wilLoul a ll~lu~ of s "u~led and nn.~ ed fatty acids.Specifically, the levels of hydrogen peroxide produced by epi(l~m~l k~r~tinocytes were measured after exposure to no ~ntioxitl~nt without fatty acids (FY~mrle 43, control) and with fatty acids (Example 44), to sodium l~y~uv~le and ascorbic acid without fatty acids (Example 45) and with fatty acids (Example 46), to sodium ~yluva~ and lactic acid without fatty acids (Example 47) and with fatty acids (Example 48), to sodium ~y~uv~le and Vitamin E without fatty acids (Example 49) and with fatty acids (Example 50), and to ascorbic acid and Vitamin E without fatty acids (Example 51) and with fatty acids (Example 52). The ability of all combin~tionQ. of ~nh~Yi-l~nt~ to reduce the hydrogen peroxide pr~lnr,tion by epid~m~l ke~tinocytes was increased in the presence of fatty acids. In order of potency, the most effective combin~ti~nQ to reduce the hydrogen peroxide production of epidermal keratinocytes were sodium pyruvate and Vitamin E, sodium pyruvate and lactic acid, and Vitamin E, each in combination with a ~lu c; of s~ ~d and nnQ~tnr~ted fatty acids (0.5%).
RecallQe of the ~;yk~tC~;Çity of cells towards ascorbic acid described above, the ascorbic acid combin~ti~n.Q without sodium ~yluvale were not c~-n.Q~ red Qignific~ntly ~li~.~,.ll from the control test solllti~n Summary Analysis Of The Data From Studies 1 and 2 Human epidermal ker~tinocytes were isolated by lly~s;~ Qn of epitheli~l sheets and grown in modified base MCDB 153 m~lillm supplen-Pntçd withepidermal growth factor and bovine piluil~y extract. Cells were seeded in culture dishes at a density of 3 x 105/dish. Prior to exposure to W B light (lOOmJ/cm2) or m~t with lOOng/ml TPA, the cultures were treated with the a~.;a~

W O96/14868 PCTrUS95/12853 concentration of wound healing coll~ ents. Tn~cell~ r prod~ ~ion of hyd~
peroxide was measured using DCFH-DA, a nonpolar com~ d that readily ~liffil~çs into cells, hydrolyzed to a nonrQl~r d~";v~live. In the ~ .sh~ce of in~Plllll~r ~ hydrogen peroxide, DCFH is oxidized to a highly fluolesce.lt compound DCF. Thus, S cellular fluorescence i~l~nsily is direc~y ~lu~,,tional to levels of hydrogen ~r~Aide produced and can be ...i.~.;(oled by flow cytometry. Hydrogen peroxide is ~;y~ic, the,~rol~, lower levels of hydrogen peroxide prod~ ion is desirable for cellularviability.

W Og6/14868 PCTnUS95/12853 In all cases, the three component wound healing composition ~Nfp?l~S~ed the p~ d o~l~...es, clearly dem~n~ n~ unpredicted ~yllc~,y.
Results 1 - Control 250 250 0 2 - Fat~,r Acids 250 230 -20 (o.5o/o) 3 ~otlinm Pyruvate 250 490 +240 (lOmM) 4 - Vitamin E 250 400 +lSO
(SO units) S - Pyruvate & 250 430 +180 Fatty- Acids lS 6 - Vitamin E & 250 200 -50 Fatty Acids 7 - Pyruvate & 250 290 +40 Vitamin E
8 - Pyruvate & 250 120 -130 Vitamin E & Fat~ Acids Column 1 shows the dirre~ t 1,~ groups.
Column 2 shows the production of H202 in control cells (fmoVcell).
Colnmn 3 shows the production of H2O2 after t~ with wound healing co,llyol~ents.
Column 4 shows the difference in pro~ll-ction of H2O2 from control after the ll&~l---~--l All comr~ Qn~ were ~ç~se~l against the controls, which produced 250 H2O2 fmol/cell. The positive numbers l~l.,se.~l H2O2 production in excess of thecontrol and the negative llul~ ers represent H2O2 pro~lnc~l.)n below the control. These results are set out in Figure 8.

-Wos6ll4868Pcr/usss/l28s3 Combination of Single In~,r~ ,..l Effects Fatty Acids (-20) & Vitamin E (+150) &~ PyluvaL~ (+240) +370 Is The Predicted Three l~omron~nt Effect -130 Is The Wound healing composition Actual Effect 500 Is The Dirr~.e.,ce Between Predicted Effect minus Actual effect (Synergy) Combination of Paired and Single Ingredients 10Pyluv~te & Fatty Acids (+ 180) & vitamin E (+ 150) +330 Is The Predicted Three Co~ ,,cnt Effect -130 Is The Wound healing composition Actual Effect 460 Is The Difference betwveen Predicted Effect minus Actual Effect (Synergy) Vitamin E & Fatty Acids (-50) & Pyruvate (+240) + 190 Is The Predicted Three Component Effect -130 Is The Wound healing composition Actual Effect 320 Is The Difference ~cLwe~ll Predicted Effect minus Actual Effect 20(Synergy) E~uv~Le & Vitamin E (+40) & Fatty Acids (-20) +20 Is The Predicted Three Component Effect -130 Is The Wound healing composition Actual Effect 25150 Is The Difference between Predicted Effect minus Actual Effect (Synergy) In all cases, the three cc,lll~ollent wound healing composition surpassed . 30the predicted ontcomes clearly ~l~ml~n~tr~ting ul~pl~dicted synergy.

-WO 96tl4868 PCT/US95/12853 E~amples Of The Therapeutic Wound Healing Compositions Of Embodiment One (LA-D) Study 3 s This study dPm- netr~tes a CO~ QI~ of the wound healing abilities of the therapeutic wound healing compositions of the present invention versus conv~ ;on~l wound healing comrosihl~n~ The results of this study are i~ tr~tecl in examples A-D.

The wound healing cc,~osiLions of Examples A-D were prepared having the compositions set out in Table A.

Fx~mpleS
Ingredient A B C D
Prep-HIM
sodium pyrubate -- 2% -- --vitarnin E -- 1%
~ic~ .n fat 2%

shark liver oil 3% 3% 3%
petrolatum in 64% 66.5% 68%
min~.r~l oil ~uull~ 22.53% 25.03% 26.8%
pA~ 11 totaling 5%
enml~ifi~r 100% 0.2% 0.2% 0.2%
$
These cullll~vl~nt~ are present in P~ ;on HTM

Wound healing composition A was co~.. ~cially available Preparation HlM. Wound healing composition B was a petrolatum base formulation co~ g live yeast cell deliv~live, shark oil, and a l~ ule of sodium ~yluv~te, vitamin E, and chicken fat. Wound healing composition C was a petrolatum base form~ tton c~ ;..;..g Iive yeast cell delivaliv~; and shark oil. Wound healing composition D was a petrolatum base formlllstti.~n only.

S Wound healing studies were carried out using h~irless mice (SKR-1, Charles River) 6-8 weeks in age. One group of mice were unll~aled as a control group and were rerelled to as Example E. In each group there were 6 mice for ev~ tion at either day 3 or day 7 for a total rlullll~e. of 60 ~nim~l~ in the study. The mice were ançstheti7çd with ether and a midline 3 cm full thir.l~n~ lonEitll~lin~l inr;~ion was made with a nu lll)cl 10 scalpel blade. Tnri~ion~ were closed using steel clips at 1 cm intervals. Formlll~ti~ A-D set out above were applied in a r~n-lomi7çd blinded study to the wounds on day 0 at 2 hours following wounding and reapplied at 24 hour intervals during the 7 days of the study. The wounds were examined daily and scored on a basis of 0-5 for closure on each day of the study, with a score of 5 r~lesrnttng the wound best healed.

The slnim~l~ were s~crifice(3 on day 3 and day 7 using cervical dislocation. The dorsal skin including the inci~ion was ~ secte~l without the subcutaneous tissue. The skin was placed in neutral buffered form~lin and subsequently sectioned and stained with k~ u~ylin and eosin. The wounds were eY~minecl rnicroscopically and represen~live tissue secti~n~ were photographed.

On each day of the eXp~rim~nt~ the score and rank order of the formlll~tions for closure of wounds and speed of healing were as follows:
B (S) >> D (4) >> C (2) >/= E, ~ontrol (2) ~ A (1) Photographs of the wounded rnice on day 4 are set out in Figures 9 and 10.

Figures 9 and 10 show that FormnlstttQn B, which was apetrolaturn base fiorrrtlll~tinn co.~ E live yeast cell d~,liv~livci, shark oil, and a l~ ule of sodium ~yluvale, vitamin E, and chicL-~n fat, was a siEnifi-c~ntly better wound healing agent than the other fonnlllsltion~ These results are su~ulled by the subjective grading of the wound closures and the speed of healing on each day (1-7) of the experirnent as well as on the objective histological e-~";~.~hi~n of tissue sectil~nC to measure the extent of infl~ m~ ,y cell infilh~te within the wound and the extent of epithe~ h~n at the wound edges. The final result was that less scar tissue was S present at day 7 on the mice treated with Fl rm~ ti~m B.
.
Formnl~h-~n D, which was a white petrolatum fiormlll~hl~n only, was judged to be cignifi~ntly more e~ilive to y,u.~ e healing than either Formnl~h~ n C, which was a petrolatum base formlll~hr~n co~ ;"g shark liver oil and live yeast cell derivative, or Formnl~tion A, which was Pl~i~ardliOn HIM. The superior ability of Formlll~h~n D over Fcrmlll~hQn C to irnprove healing may result from a delay in the healing process caused when the live yeast cell de~ivaLive is depleted and the cells shift to an ~ltern~tive m~hient source. The presence of the lY~lwe of sodiurn pyruvate, vitarnin E, and chicken fat in Form~ tir~n B aypd~ ly offsets the depletion of the live yeast cell dc,;valive.

Formlll~hon C, which was a petrolatum base formnl~hQn co~ ;..i..g live yeast cell derivative and shark oil, was judged comparable to the conhrol (w~tleated wound) in speed of wound closure and extent of h~ling Formnl~h-~n A, which was P~a~ion HTM, appeared to be the least effective healing fc~rmlll~h~n by both subjective grading of wound healing and by objective eY~min~hon of tissue sech~nc.
The superior ability of F~rmlll~h-~n D and Formlll~hon C over FQrmlll~hon A to improve healing may be due to their ability to act as an occlusive wound dressing that pl'eVWll5~ t-ransepiderrnal water loss and thus pl~lll;)teS healing and wound closure. The poor ability of Formlll~h~n A to illlyl~ve healing may be due to the potential Cy~ ;City of phenyh.lcr~;wic nihrate present in Prep~r~h~n HIM as a pres~.v~live.
.

These results show that the wound healing compositions of the present invention which comprise a ~ we of sodium pyluv-dle, vitamin E, and chicken fat increase the proliferation and resll.cc.it~ti~n rate of ".i1.""~ n cells. The wound healing compositions me~ te low levels of oxygen in the initial stages of healing to :iUyylC;Ss oxidative damage and higher levels of oxygen in the later stages of healing to ylolll~l~ collagen form~h~n Il. Acne Treating-Wound Healing Compositions A. Embodiment Two ~A-D ~ X) Applicant has discovered t~l~apc.lLic acne treating-wound healing amrosihon.~ (IA-D + X) which cnmrri~e tretinoin (X) and the wound healing composihon~ of Embo~limto.nt One a.A-D). Preferably, the wound healing composition (IA) co...l..;aes (a) pyruvate, (b) an ~ntir)xi-l~nt, and (c) a mi~lul~ of s.,~ ~ and unsalu,~lt;d fatty acids. Tretinoin is useful for the topical ~ f..~ of acne vulgaris but is known to induce excessive redness, ed~ .us bli~tering or crusting, and severe local erythema and peeling at the site of applic~hc n Wound healing compositi~n~ can increase the resll~c;l~;ol- rate of injured l,.AI,~ AliAn cells and the proliferation rate of new ~... AliAn cells to replace dead cells but do not treat acne vulgaris. Applic~Ant~
have found that the colllbin~lion of tretinoin and a wound healing composition results in a therapeutic acne treating-wound healing composition which reduces the duration and severity of acne vulgaris, as an a lg",~ A wound healing composition, and the i1ThAtion associated with tretinoin, as a cyk~l~clive-wound healing composition.This invention also relates to mfth~l~ for employing the therapeutic acne treating-wound healing compositions to treat wr;nkles.

The combination of tretinoin and the wound healing composihon~ of the present invention provides a phArmAcellticAl composition useful for treating acne vulgaris and having an enhAnce~l ability to prevent and reduce injuTy to ".~ ...AliAn cells and flrther increase the rf~ ,sn~citAti~ln rate of injured IllAllllllAliAn cells. The tissue damage ~A~soci~ted with tretinoin in~ cçfl inflAmmAhon is believed to be caused by the pro~lllction of cellular produced active oxygen species. Combination of tretinoin and the wound healing compositions may ~u~less such reactive oxygen-linked tissue injury.

As set out above, tretinoin (Retin-AlM) is in-licAt~cl for the topical 1~ l of acne vulgaris. Ch~ micAlly~ tretinoin is all-trans-retinoic acid (vitamin A
acid). Tretinoin is available in a gel vehicle con~i~hng of butylated hyd~ y~h7~nç, h~ AylJl~)yl celllllose and alcohol; in a hydrophilic cream vehicle con~i~hng ofstearic acid, iso~r~y-l myristate, polyoxyl 40 stÇAr~te~ stearyl alcohol, ~AnthAn gum, sorbic acid, butylated h~ ~luene, and l,~iLed water, and in a liquid vehicle C~ Q~;-.g of polyethylene glycol 400, butylated l,~ohl~ne, and ~k,Qhol Tl~linoin rnay be used in many distinct physical forms well known in the pk-....~G~II;c~l art to provide an initial dosage of tretinoin and/or a fur~er time-release fonn of tretinoin. Without being limited thereto, such physical forms include free folms and encapsulated forms, and IlliA~ S thereof.

The amount of l,~,lino", used in the present ill~,cl,lioll may vary depending upon the t~l~e.llic dosage recQ.. - .~ed or p~ lr~l for the particularpatient. In general, the amount of tretinoin present is the ordinary dosage required to obtain the desired result. Such dosages are known to the skilled practitioner in the medical arts and are not a part of the present invention. In a ~,er~ ;d embo~lim~nt~
tretinoin in the acne treating-wound healing composition is present in an amount from about 0.01% to about 0.1%, and preferably from about 0.025% to about 0.05%, by weight.

B. Methods For Making The Acne Treating-Wound Healing Compositions Of Embodiment Two (I.A-D I X) The present invention extends to mçth~3c for making the thcl~c.llic acne treating-wound healing compositions (I.A-D + X). In general, a thc~a~ lic acne treating-wound healing composition is made by forming an ~d~ e of the wound healing components of Embodiment One (I.A-D) and l~c;l~lOill. In a first aspect of Fmbo~lim~nt Two (I.A + X), an acne treating-wound healing therapeutic ccmposi1~on is made by forming an a~ lu.e of tretinoin and a wound healing composition comprising (a) a pyruvate, (b) an antioxidant, and (c) a mixture of saturated and unsalulaled fatty acids. In a second aspect of Embodiment Two (I.B + X), an acnetreating-wound healing therapeutic composition is made by forming an ad~lule of tretinoin and a wound healing cum~o~ilion cQmrn~ing (a) a pyruvate, (b) a lactate, and (c) a mi~lule of s~.lulaled and lm~tll~ted fatty acids. In a third aspect of Embo lim~nt Two (I.C + X), an acne treating-wound healing therapeutic composition is made by W O96/14868 . PCTrUS95/12853 fqrminE an a~l...;x~ e of tretinoin and a wound healing composition co~ ;ci~.g (a) an ~nt~oYi~l~nt~ and (b) a ll~i~luÇ~ of s~ cl and l~c~ nl~1 fatty acids. In a fourth aspect of Embodiment Two a.D + X), an acne treating-wound healing th~ ;c composition is made by fqrmin~ an a~l...;xl~ of t-retinoin and a wound healing S cqmrocit l n cQmrricinE (a) a lactate, (b) an ~ntioYi-l~nt, and (c) a ll~i~ of s and unc~tllr~tecl fatty acids.

In a ~l~rt;ll~d embo~im~nt the invention is dilectt;d to a m~thod for preparing a therapeutic acne treating-wound healing composition a.A + X) which c~ mrrices the steps of a~lmixinE the following ingredients:
(A) a therapellt ç~lly t;rrteLive amount of tretinoin; and (B) a wound healing cqmposihQn which comrrices (a) pyruvate selected from the group con.cictin~ of pyruvic acid, ph~Tm~ceutic~lly acceptable salts of pyruvic acid, and llli~lulc;s thereof;
(b) an ~nhoxi~l~nt and (c) a ~ lule of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular lll~,~I,l~les and resusciL~Li-", of . . .i. . .~. . .~li~n cells.

C. Methods For Employing The Acne Treating-Wound Healing Compositions of F~n'o liment Two (I.A-D ~ X) I

The present invention extends to methods for employing the th~ c ~l;c acne treating-wound healing compositions ~l.A-D + X). In general, a therapeutic composition is employed by cont~cl;nE the therapeutic composition with acne vulgaris.
In a pl.,Çt;lled embo~lim~nt, the invention is directed to a method for treating acne vulgaris in a human with an acne treating-wound healing composition (I.A + X) which comrrice~C the steps o~
(A) providing a therapeutic acne treating-wound healing composition which comprises:
(1) a therapeutically effective amount of tretinoin; and .
CA 02204l4S l997-04-30 W 096/14868 PCT~US95/12853 (2) a wound healing coll,posilion which cc....~ e~
(a) ~yluv~lv s~kvl~d from the group c~n~;~t~ng of pyruvic acid, ph~rm~ce11tic~11y acceptable salts of pyluvic acid, and ~LulCS t_ereof;
(b) an ~nt1~xirl~nt and (c) a~ e of s~ l andnn~ fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular ..,r"~ ol~çs andresn~ç;l~1iol- of..,~ n cells; and (B) cont~ct ng the acne treating-wound healing composition with the infected wound.
In another ~lef~,,lvd embo~lim~.nt, the present invention extends to mçtho-l~ for employing the the ~e.lLic acne treating-wound healing co~ osilions(I.A-D + X) to treat wrinkles. Tretinoin is also used to decrease wrinkles because itdecreases the cohesiveness of follicular epithelial cells with decreased microcomedo fc-rm~tion In general, a therapeutic composition is employed by c~nt~cting the therapeutic composition with the wrinkles. In a specific embodim~-nt, the invention is directed to a method for treating wrinldes in a human with an acne treating-wound healing composition (l.A-D + X) which compri~es the steps of:
(A) providing a therapeutic acne treating-wound healing composition which comprises:
(l) a th~r~pellhc~lly effective amount of tretinoin; and (2) a wound healing composition cu~ g (a) pyruvate s~lecte~l from the group con~ ting of pyruvic acid, ph~rm~ce11tic.~11y acceptable salts of pyruvic acid, and ~ vs thereof;
(b) an ~ntioxi~l~nt and (c) a ~ e of s~ e~1 and nn~tl1r~te(1 fatty acids wherein the fatty acids are those fatty acids required for the repa* of cellular ~ es and res1~ ;on of ",~ "~ n cells; and (B) cont~cting the acne treating-wound healing composition with the wrinkles.
The types of wounds which may be healed using the acne treating-wound healing compositions and the ~ngJ..f.~ecl acne treating-wound healing co~ o~ iolls of the present invention are acne vulgaris.

WO 96/14868 PCrlUS95/12853 Mçthods for treating acne vulgaris or wrinkles c-....l..;~e topically ~dmini~t~ing the compositions of dle present ~lv~ Lion direcdy to the acne vulgaris or wrinkles. The cQmrositic-n is .~;n~ ecl in contact with the acne vulgaris or - wrinkles for a period of tirne sllffic;~nt to increase the proliferation and recll~cit~ti~n S rate of the cells.

D. Augmented Acne Treating-Wound Healing Compositions Of Embodiment Two ~.A-D I X I M) In another aspect of Embo.lim~nt Two, the therapeutic acne treating-wound healing CO111~0SjliOnS (I.A-D + X) of dle present invention may be furthercombined widl m~li~ useful for treating wounds (M) to form allg,--f nlecl acne treating-wound healing compositions (I.A-D + X + M). In this emb~limPnt, the combination of the acne treating-wound healing compositif)n of the present invention and the meclic~mt~nt useful for treating wounds provides an a~lg---' I.t~1 acne treating-wound healing composition having an enhanced ability to increase the proliferation and resll~cit~hon rate of ""~"",~ n cells. For example, the therapeutic compositiQn~ of the present invention may be used in cul~ ation with m~lic ~~..c .l~ useful for treating wounds such as immunoshm~ hng agents (BetafectinTM), antiviral agents, antikeratolytic agents, anti-inll~ agents, ~nhfilngal agents, tretinoin, sunscreen agents, d.orm~tological agents, topical antihiFt~mine agents, ~nhbact~ri~l agents, bio~-lh~ive agents, re~i~ y bursting inhibitors (lactic acid, ad~nosine), inhibitors of prost~gl~n.lin synthesis (ibu~r~rcll, aspirin, indometh~c.in, meclofenomic acid, retinoic acid, padim~te O, meclomen, oAyl~el~olle), steroidal anti-infl~ ol~ agents (corticosteroids including synthetic analogs), ~nhmir.robial agents (neosporin o;..l~
silvadine), antiseptic agents, ~nes~çhc agents (~l~luAille hydrochl-)ri~e, lidocaine, ben7~c~ine), cell nutrient media, burn relief medic~hQn~, sun burn mPAir~ti~n.~ acne l;o...~, insect bite and sting mPAic~hon~ wound cle~n~ers, wound dr s~ingsJ scarre(ll~c.ing agents (vitamin E), and the like, and ll~ ules thereof, to further enhance the proliferation and resnscit~hon rate of ~ .".~,Ali~n cells. Preferably, the m~Aic~m~ nt useful for treating wounds is selectecl from the group cQn~i~tinp of immllnostlm~ ting agents, antiviral agents, antikeratolytic agents, anti-;..ll~ agents, antifungal CA 02204l4S l997-04-30 W O96/14868 PCTrUS95/12853 agents, llcli~ Dwls~,lce.l agents, df~m~tnlogical agents, topical ~ntil~ .e agents, ~nt bactpri~l agents, bio~lhssive agents, ltD~ oly bursting inhibitors, inhibitors of prost~gl~n(1in synthesis, ~ntimi~robial agents, cell mltnpnt media, scar redu~in~ agents, and llfi~lw~cs thereof. More preferably, the me~ ment useful for treating wounds is S s~lGct~d from the group concicting of immlmoshmlllat1ng agents, antiviral agents, ~ntikPr~tolytic agents, anti-inflS~ y agents, antifungal agents, acne treating agents, sunscreen agents, df rm~tological agents, ~ntihicpmine agents, antibacteri~l agents, bioa-lhecive agents, and l~ lwcs thereof.

In a pl~;rcllcd embo-1imP,nt, the invention is d-lcclcd to an all~ ed acne treating-wound healing composition a.A + X + M) which comprises:
(A) a thel~l.c.~lic acne treating-wound healing composition which comrri.cç.c (1) a therapeutically effective amount of tretinoin; and (2) a wound healing composition which comprises:
(a) pyruvate selectf,d from the group co~cictln- of pyruvic acid, ph~rm~ceutic~lly acceptable salts of pyruvic acid, and mi~lulcs thereof;
(b) an antioxidant; and (c) a ll~i~lulc of salul~lcd and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular ~ f,c and rf,snccit~tion of ~ n cells; and a3) a m~lic~mP.nt useful for treating wounds.

The present invention extends to methods for making the all~ f ~led acne treating-wound healing compositionc In general, the anp.m~nted compositions are made by ~mixing the therapeutic acne treating-wound healing composition with themedic~mPnt useful for treating wounds to prepare the a~ .lt;d acne treating-wound healing composition.

The present invention extends to mP,tho lc for employing the therapeutic ~ngmP.ntfed acne treating-wound healing compo~citic-nc In general, a therapeuticallg~,. ,l~d composition is employed by cont~cting the therapeutic composition with the acne vulgaris. In a pl~crcllcd embo-limPnt, the invention is directed to a method for treating acne vulgaris in a human with an ~n~mpntsd acne treating-wound healing cQmrositi~ n acne treating-wound healing comrositi-~n (I A + X + M~ which co~ ;ces the steps of:
(A) providing a therapeutic allgmPntP~ acne treating-wound healing composition which ct~mrrices:
(1) a therapeutically effective amount of tretinoin;
(2) a wound healing composition which comrices (a) pyruvate selected from the group conQicting of pyruvic acid, ph~rm~eelltic~lly acceptable salts of pyruvic acid, and ~ ul'dS thereof;
(b) an ~ntiox~ nt and (c) a llfi~Lult; of sa~ aldd and nn.c~hlr~ted fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular llltl~ es and resllccit~iQn of ~ n cells; and (3) providing a mP11ic~mPnt useful for treating wounds; and (B) cont~cting the acne treating-wound healing composition with the acne vulgaris.

In another ~;rt;lled embo.1imPnt, the present invention extends to mPthoflc for employing the therapeutic ~llgmPnte(l acne treating-wound healing compo,citi~nC to ~¢eat wrinkles. In gPnPr~l a therapeutic ~lgmPntPd composition is employed by cont~chng the th~ ; c.lLiC composition with the wrinkles. In a specific embo-limPnt the invention is directed to a method for treating wrinkles in a human with an ~ngmpnted acne treating-wound healing co~l~osiLion (I.A + X + M) which compricP,s the steps of:
(A) providing a therapeutic acne treating-wound healing composition which c~.. l.. ;ce~
(1) a thP.r~re~lti~lly ~rre~;Live amount of tretinoin; and (2) a wound healing composition comrricing (a) pyruvate selected from the group wnQ;ctin,P~ of pyruvic acid, ph~rm~celltically acceptable salts of pyruvic acid, and llfi~ es thereof;
(b) an ~ntiQxid~nt and (c) a ll~Lult; of s~ 1 and lmc~hlr~tP,(l fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular ~tl~ es and rP,s~lsc.it~tion of ,~,~"""Ali~n cells; and W O96/14868 PCTrUS95/12853 ~ (B) cnnt~cting the acne treating-wound healing com~o~ilion with the wrinkles.

E. Formulations Of The Acne Treating-Wound Healing Compositions S Of Embodiment Two aA-D I X) and aA-D I X I M) Once prepared, the i~v~ live therapeutic acne treating-wound healing compositions and ~ng~n. -led acne treating-wound healing comrocihonc _ay be stored for future use or may be form~ ted in errc;~;live ~luull~ with ph~rm~celltieallyacceptable carriers such as ph~rm~t~.ellti~l appli~ncec and topical vehicles (oral and non-oral) to prepare a wide variety of rh~rm~ce~tic~l compocitionc The ph~rm~centiç~lly acceptable carriers which _ay be employed and the mtoth~c used to prepare the ph~rm~cellhcal compositions have been described above in connection with the fo~nlll~hons of the wound healing compositions of Embodiment One (I.A-D).
In a ~rercllcd embodiment, the invention is dilccled to an acne hreating-wound healing ph~rm~ce~lhc~l composition which comprices (A) a the,~c.llic acne hreating-wound healing composition a.A + X) which comprises:
(1) a therapeutically effective amount of tretinoin; and (2) a wound healing composition which comrrices (a) pyruvate s~lected from the group con.cichng of pyruvic acid, ph~rm~eellhcally acceptable salts of pyruvic acid, and ~ S thereof;
(b) an antioxidant; and (c) a IlliX.IlllC of s~ led and nnc~hlr~tçd fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular .. ~ çs and reCllc~it~hon of ,..~I "",~ n cells; and (B) a ph~rm~cell~ic~lly acceptable carrier selected from the group conQi~ting ofph~rm~cel1hc~1 appliances, bio~tlhecives~ and occlusive vehicles.

In another ~lcrt;llcd embotlim~-nt, the invention is directed to a method for pl~illg a ph~rm~cel~hic~l composition for increasing the prolifer~hon and resllccit~hc!n rate of "~i1."".~ n cells, which comrricels the steps of:

W O96/14868 PCTnUS95112853 (A) providing a ~c~ ically eLr~live am~7~lnt of an acne treating-wound healing co.,ll)o~iLion a A + X) which comrn~es (1) tretinoin; and (2) a wound healing composition co...l..;~.,l~
S (a) pyruvate selecte~,7 from the group con~ in~ of pyruvic acid, ph~rm~cellt c~lly acceptable salts of pymvic acid, and ~lules thereof;
(b) an ~mt7r)xi~7~nt, and (c) a ~lu.e of satm~t~7, and lm~7tnr~7ted fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of l~ "~ n cells;
(B) providing a ph~nn~ce~lt7c~11y acceptable carrier; and (C) a-7miXi71g ~e acne treating-wound healing composition from step (A) and the ph~ ceutically acceptable carrier from step (B) to form a ph~rm~ce~ltic~l composition.

Claims (20)

I claim:

ACNE TREATING-WOUND HEALING COMPOSITIONS

1. A therapeutic antiacne-wound healing composition for the topical treatment of acne vulgaris which comprises a therapeutically effective amount oftretinoin and a wound healing composition, wherein the wound healing compositioncomprises:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) antioxidant and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the resuscitation of injured mammalian cells.

2. The composition according to claim 1, wherein the pyruvate is selected from the group consisting of pyruvic acid, lithium pyruvate, sodium pyruvate, potassium pyruvate, magnesium pyruvate, calcium pyruvate, zinc pyruvate, manganese pyruvate, methyl pyruvate, .alpha.-ketoglutaric acid, pharmaceutically acceptable salts of pyruvic acid, prodrugs of pyruvic acid, and mixtures thereof.

3. The composition according to claim 2, wherein the pyruvate is sodium pyruvate.

4. The composition according to claim 1, wherein the antioxidant is selected from the group consisting of all forms of Vitamin A including retinol and 3,4-didehydoretinol, all forms of carotene including .alpha.-carotene, .beta.-carotene, gamma-carotene, and delta-carotene, all forms of Vitamin C including D-ascorbic acid and L-ascorbic acid, all forms of Vitamin E including .alpha.-tocopherol, .beta.-tocopherol, gamma-tocopherol, delta-tocopherol, tocoquinone, tocotrienol, Vitamin E esters which readily undergo hydrolysis to Vitamin E including Vitamin E acetate and Vitamin E succinate, and pharmaceutically acceptable Vitamin E salts such as Vitamin E phosphate, prodrugs of Vitamin A, carotene, Vitamin C, and Vitamin E, pharmaceutically acceptable salts of Vitamin A, carotene, Vitamin C, and Vitamin E, and mixtures thereof.

5. The composition according to claim 4, wherein the antioxidant is Vitamin E acetate.

6. The composition according to claim 1, wherein the mixture of saturated and unsaturated fatty acids is selected from the group consisting of animal and vegetable fats and waxes.

7. The composition according to claim 6, wherein the mixture of saturated and unsaturated fatty acids is selected from the group consisting of human fat, chicken fat, cow fat, sheep fat, horse fat, pig fat, and whale fat.

8. The composition according to claim 7, wherein the mixture of saturated and unsaturated fatty acids comprises lauric acid, myristic acid, myristoleic acid, pentadecanoic acid, palmitic acid, palmitoleic acid, margaric acid, margaroleic acid, stearic, oleic acid, linoleic acid, linolenic acid, arachidic acid, and gadoleic acid.

9. The composition according to claim 1, wherein tretinoin is present in the therapeutic wound healing composition in an amount from about 0.01% to about 0.1%, by weight of the therapeutic wound healing composition.

10. The composition according to claim 1, wherein pyruvate is present in the therapeutic wound healing composition in an amount from about 10% to about 50%, by weight of the therapeutic wound healing composition.

11. The composition according to claim 1, wherein the antioxidant is present in the therapeutic wound healing composition in an amount from about 0.1%
to about 40%, by weight of the therapeutic wound healing composition.

12. The composition according to claim 1, wherein the mixture of saturated and unsaturated fatty acids is present in the therapeutic wound healing composition in an amount from about 10% to about 50%, by weight of the therapeutic wound healing composition.

13. A method for treating acne vulgaris in a human with an acne treating-wound healing composition which comprises the steps of:
(A) providing a therapeutic acne treating-wound healing composition which comprises:
(1) a therapeutically effective amount of tretinoin, and (2) a wound healing composition comprising (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof, (b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and (B) contacting the acne treating-wound healing composition with the acne vulgaris.

14. A method for treating wrinkles in a human with an acne treating-wound healing composition which comprises the steps of:
(A) providing a therapeutic acne treating-wound healing composition which comprises:
(1) a therapeutically effective amount of tretinoin; and (2) a wound healing composition comprising:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and (B) contacting the acne treating-wound healing composition with the wrinkles.

\

15. A method for preparing a therapeutic acne treating-wound healing composition for the topical treatment of acne vulgaris which comprises the steps of admixing the following ingredients:
(1) a therapeutically effective amount of tretinoin; and (2) a wound healing composition comprising:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells.

16. An augmented antiacne-wound healing composition which comprises:
(A) a therapeutic antiacne-wound healing composition which comprises:
(1) a therapeutically effective amount of tretinoin; and (2) a wound healing composition which comprises:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and (B) a medicament useful for treating wounds.

17. The augmented antiacne-wound healing composition according to claim 16, wherein the medicament useful for treating wounds is selected from thegroup consisting of immunostimulating agents, antiviral agents, antikeratolytic agents, anti-inflammatory agents, antifungal agents, other acne treating agents, sunscreen agents, dermatological agents, antihistamine agents, antibacterial agents, bioadhesive agents, respiratory bursting inhibitors, inhibitors of prostaglandin synthesis, antimicrobial agents, antiseptic agents, anesthetic agents, cell nutrient media, burn relief medications sun burn medications, insect bite and sting medications, wound cleansers, wound dressings, scar reducing agents, and mixtures thereof.

18. A method for treating acne vulgaris in a human with an augmented antiacne-wound healing composition which comprises the steps of:
(A) providing a therapeutic augmented antiacne-wound healing composition which comprises:
(1) a therapeutically effective amount of tretinoin;
(2) a wound healing composition which comprises (a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant; and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and (3) a medicament useful for treating wounds; and (B) contacting the augmented antiacne-wound healing composition with the acne vulgaris.

19. A method for treating wrinkles in a human with an augmented antiacne-wound healing composition which comprises the steps of:
(A) providing a therapeutic augmented antiacne wound healing composition which comprises:
(1) a therapeutically effective amount of tretinoin;
(2) a wound healing composition which comprises;
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and (3) a medicament useful for treating wounds; and (B) contacting the augmented antiacne-wound healing composition with the wrinkles.

20. An antiacne-wound healing pharmaceutical composition which comprises (A) a therapeutic antiacne-wound healing composition which comprises:
(1) a therapeutically effective amount of tretinoin; and (2) a wound healing composition which comprises:
(a) pyruvate selected from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid, and mixtures thereof;
(b) an antioxidant, and (c) a mixture of saturated and unsaturated fatty acids wherein the fatty acids are those fatty acids required for the repair of cellular membranes and resuscitation of mammalian cells; and (B) a pharmaceutically acceptable carrier selected from the group consisting of pharmaceutical appliances, bioadhesives, and occlusive vehicles.