(12) United States Patent
`N’Guyen et al.
`
`US006316012B1
`US 6,316,012 B1
`Nov. 13, 2001
`
`(10) Patent N0.:
`(45) Date of Patent:
`
`(54) COSMETIC OR PHARMACEUTICAL
`COMPOSITION COMPRISING, IN
`COMBINATION, A PEROXIDASE AND AN
`ANTI-SINGLET OXYGEN AGENT
`
`(75) Inventors: Quang lan N’Guyen, Antony;
`Christian Colin, Paris, both of (FR)
`
`(73) Assignee: L’Oreal, Paris (FR)
`
`* N'
`ot1ce:
`
`Sbj
`yd'l'
`h
`fh'
`u ect to an 1sc a1mer, t e term 0 t is
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 0 days.
`
`(21) Appl. No.: 09/413,496
`(22) Filed:
`Oct. 6, 1999
`
`Related US. Application Data
`
`(63) Continuation of application No. 08/923,398, ?led on Sep. 4,
`1997, now abandoned, which is a continuation of application
`No. 08/476,060, ?led on Jun. 7, 1995, now abandoned,
`which is a continuation of application No. 08/109,266, ?led
`on Aug. 20, 1993, now abandoned.
`Foreign Application Priority Data
`
`(30)
`
`Sep. 1, 1992
`
`(FR) ................................................ .. 92 10439
`
`(51) Int. Cl.7 ................................................... .. A61K 38/43
`(52) us. Cl. ....................... .. 424/401; 424/78.02; 424/59;
`424/94.1; 514/937; 514/844
`(58) Field of Search ........................ .. 424/401, 59, 78.02,
`424/94.1; 514/844, 937
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`12/1978 Kalopissis.
`4,129,644
`9/1984 Rosenbaum et al. .
`4,473,550
`9/1986 Herlihy.
`4,609,544
`9/1990 Kross.
`4,956,184
`6/1991 N’Guyen.
`5,023,235
`9/1993 Junino.
`5,244,497
`11/1994 Zysman.
`5,362,494
`12/1995 Marion et al. .
`5,474,777
`5,686,082 * 11/1997 N’Guyen ............................ .. 424/401
`
`FOREIGN PATENT DOCUMENTS
`
`0307626
`0397227
`2112550
`2315991
`2378796
`
`3/1989
`11/1990
`6/1972
`1/1977
`8/1978
`
`2400358
`2400359
`2416008
`02620024
`0111411
`03-236320
`05-124980
`87/07838
`88/02600
`91/06639
`92/01466
`
`3/1979 (FR).
`3/1979 (FR).
`8/1979 (FR).
`3/1989 (FR).
`8/1980 (JP).
`10/1991 (JP).
`10/1991 (JP).
`12/1987 (WO).
`4/1988 (WO).
`5/1991 (WO).
`2/1992 (WO).
`
`OTHER PUBLICATIONS
`
`Igarashi, Osamu, “Vitamin E Focusing on Antioxidative
`Activity, Biological Activity of Vitamin E Analogues, Trans
`portation in Blood, and Distribution in Tissues and Organs,”
`Institute of Environmental Science for Human Life, 1991.
`Database WPIL Week 8943, Derwent Publications Ltd.,
`London, GB; AN 89—310020 & DD—A—268 157.
`D.L. Bissett et al., “Protective Effect of Topically Applied
`Conjugated Hexadienes Against Ultraviolet Radiation—In
`duced Chronic Skin Damage in the Hairless Mouse,” Pho
`toderm. Photoimmunol. Ph0t0mea'., 1990: 7: pp. 63—67.
`D.L. Bissett et al., “Photoprotective Effect of Superoxide—S
`cavenging Antioxidants Against Ultraviolet Radiation—In
`duced Chronic Skin Dasmage in the Hairless Mouse,”
`Photoderm. Photoimmunol. Ph0t0mea'., 1990: 7: pp. 56—62.
`American Health, Jul/Aug. 1994, p. 72.
`
`* cited by examiner
`
`Primary Examiner—Raj Bawa
`(74) Attorney, Agent, or Firm—Oliff & Berridge, PLC
`(57)
`ABSTRACT
`
`A method of preventing damage caused to healthy skin, hair
`or mucous membranes by free radicals, includes applying to
`the healthy skin, hair or mucous membrane a composition
`containing at least one product having a peroxidase activity
`capable of reducing organic peroxides and at least one
`antioxidant capable of neutralizing singlet oxygen, the com
`position being free of peroxide. The product having a
`peroxidase activity is present in an amount of from 0.005 to
`5.0% by weight, and the antioxidant is present in an amount
`of from 0.005 to 3.0% by weight, with a weight ratio of the
`product having a peroxidase activity to the antioxidant of
`from 0.001 to 0.3. The product having a peroxidase activity
`capable of reducing organic peroxides is lactoperoxidase,
`and the antioxidant is selected from quinolines, polyphenols,
`and carotenoids.
`
`11 Claims, No Drawings
`
`L'OREAL USA, INC. EX. 1013
`
`1 of 9
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`

`

`US 6,316,012 B1
`
`1
`COSMETIC OR PHARMACEUTICAL
`COMPOSITION COMPRISING, IN
`COMBINATION, A PEROXIDASE AND AN
`ANTI-SINGLET OXYGEN AGENT
`
`This is a Continuation of application Ser. No. 08/923,398
`?led Sep. 4, 1997 noW abandoned, Which in turn is a
`Continuation of application Ser. No. 08/476,060 ?led Jun. 7,
`1995 noW abandoned, Which in turn is a Continuation of
`application Ser. No. 08/109,266 ?led Aug. 20, 1993 noW
`abandoned. The entire disclosures of the prior applications
`are hereby incorporated by reference herein in their entirety.
`The subject of the invention is a synergistically acting
`cosmetic or pharmaceutical composition containing, in
`combination, a peroxidase and an antioxidant
`Specialists currently consider that one of the causes of
`cellular aging is the reduction in the defence capacities
`against free radicals and against the oxidation phenomena
`(especially the formation of peroxides) Which they initiate.
`It is knoWn moreover that the toxicity of atmospheric
`pollutants, especially gaseous pollutants such as sulphur
`dioxide, oZone and nitrogen oxides, is linked especially to
`their free radical-initiating activity, source of oxidation
`phenomena Which cause, in living beings, cellular damage.
`Living cells, Which are in direct and permanent contact
`With the external medium (especially the skin, the scalp and
`certain mucous membranes), are particularly sensitive to
`these effects of gaseous pollutants, Which result especially in
`an accelerated aging of the skin, With a complexion lacking
`brightness and a premature formation of Wrinkles or small
`Wrinkles, and also in a decrease in the vitality and a dull
`appearance of the hair.
`It is also knoWn that the irritation phenomena caused by
`exposure to ultraviolet rays also lead to the phenomenon of
`accelerated cellular aging, and are currently considered as a
`factor for inducing skin tumours. The irritation caused by
`UV radiation gives rise, in this case as Well, to the formation
`of radical species Which lead especially to the oxidation of
`skin lipids, and it is thought that lipid peroxides are one of
`the factors Which trigger photocarcinogenesis. It is knoWn in
`particular that the induction of ornithine decarboxylase
`(abbreviated ODC) constitutes an early marker for skin
`tumour, and that organic peroxides are capable of inducing
`the formation of ODC in the epidermis; see R. L. Binder et
`al., Carcinogenesia, Vol. 10, No.12, 2351—2357 (1989).
`Living cells possess various natural means of defence
`against lipid peroxides, in particular epidermal glutathione
`peroxidase, but the effectiveness of the detoxifying activity
`of the latter is substantially decreased under the in?uence of
`an exposure to ultraviolet radiation.
`It in therefore important to develop active systems Which
`make it possible to combat the harmful effects of peroxides,
`especially the organic peroxides formed under the action of
`atmospheric pollutants and ultraviolet radiation.
`It is knoWn that some antioxidant are capable of confer
`ring a protection against the skin damage caused by radia
`tions or peroxides, including When theme antioxidants are
`applied topically; see for example Bissett at al., Photoderm.
`Photoimmunol. Photomed. 7,56—62 and 63—67 (1990).
`By studying certain antioxidant systems and by using the
`induction of ODC as marker, the Applicant has discovered
`that, surprisingly, certain combinations had the property of
`inhibiting the formation of ODC While the constituents of
`the combination, When used alone, had no effect or even
`caused an increase in the induction of ODC.
`It has been discovered more precisely that the peroxi
`dases capable of reducing organic peroxides cause an
`
`2
`increase in the induction of ODC by ultraviolet radiation,
`and that certain antioxidant are Without signi?cant effect on
`the induction of ODC. Such in the came especially for the
`antioxidant. capable of neutraliZing singlet oxygen, Which
`are therefore anti-singlet oxygen agents. It has hoWever been
`discovered that the combination of peroxidases capable of
`reducing organic peroxides With antioxidants capable of
`neutraliZing singlet oxygen, makes it possible to substan
`tially neutraliZe the induction of ODC. Such a combination
`therefore has synergistic properties.
`These useful properties can be exploited by incorporating
`such synergistic combinations into cosmetic or pharmaceu
`tical compositions in a form Which alloWs application to the
`skin, super?cial body groWths and mucous membranes.
`The subject of the invention is therefore a cosmetic or
`pharmaceutical composition characteriZed by the fact that it
`comprises, in combination, at least one product having a
`peroxidase activity capable of reducing organic peroxides
`and at least one antioxidant capable of neutraliZing singlet
`oxygen.
`The composition of the invention in an antioxidant
`composition Which therefore does not contain peroxide. In
`particular, it does not contain hydrogen peroxide.
`There may be used as product having a peroxidase
`activity any substance capable of reducing organic peroxides
`in the presence of an electron donor.
`These peroxidases may be especially peroxidases of
`natural (plant or animal) origin, or alternatively peroxidases
`modi?ed chemically or by grafting, by adsorption onto
`supports or by encapsulation (see for example applications
`PCT WO 87/07838 and EP-A-0,397,227).
`There may be used especially lactoperoxidases, fungal
`microperoxidases, myoloperoxidase and the like.
`It in knoWn that lactoperoxidase (abbreviated LPO) is an
`enzyme Which occurs especially in numerous mammalian
`tissues and secretions, Which uses one of the numerous
`cellular electron donors to reduce organic peroxides of the
`ROOH typo (R being an organic group). Lactoperoxidase is
`a commercial product, sold especially by the companies
`Sigma and Sederma.
`There may be also used recombinant peroxidases, for
`example recombinant LPO (Patent Application Wo
`91-06639).
`The antioxidant capable of neutraliZing singlet oxygen is
`chosen especially from quinoline and its derivatives,
`polyphonols, carotenoid derivatives and nucleosides and
`their derivatives.
`Among the quinoline derivatives Which can be used,
`there may be mentioned in particular 6-athoxy-1,2-dihydro
`2,2,4-trimethylquinoline or ethoxyquine, in the form of a
`monomer, dimer or oligomer or mixtures of these various
`forms, and ethoxyquine derivatives.
`There may be used in particular the ethoxyquine deriva
`tives of formula (I)
`
`(1)
`
`10
`
`15
`
`25
`
`35
`
`45
`
`55
`
`65
`
`in Which A represents a group —CO—B,
`B representing especially a group (CHOR‘)nR in Which R‘
`represents a hydrogen atom, an acyl, alkyl or aralkyl
`group,
`
`2 of 9
`
`

`

`US 6,316,012 B1
`
`F.
`
`<11)
`
`(III)
`
`10
`
`or (III):
`
`15
`
`3
`R is a hydroXymethyl, carboXyl, carboXyalkyl,
`carboXyaryl, carboXyarylalkyl, carboXamide or
`—COR‘“ group, R‘“ representing ethoXyquine residue
`(formula (I) less the substituent A) or R‘“ representing
`—CRZOR“, R“ being an acyl, aralkyl or alkyl radical
`and n being an integer from 2 to 6;
`or B represents a group —(CHR““)mN+R1R2R3X_ in
`Which R““ represents a hydrogen atom, an optionally
`substituted heterocyclic, aralkyl, aryl or alkyl radical,
`or R““ represents —(CH2)qCOOH Where q is a number
`Which may vary from 1 to 3, R1, R2 and R3 indepen
`dently representing a hydrogen atom, an aryl radical, a
`heterocyclic aryl radical, a substituted or unsubstituted
`cycloalkyl or alkyl radical, X“ being an anion and m
`being an integer from 1 to 6;
`or B represents a group —(CHR““)m NR1R2, Where R““,
`R1, R2 and m have the meanings stated above;
`or B represents a group NR‘lR‘2 in Which R‘1 and R‘2
`independently represent a substituted or unsubstituted
`aryl, heterocyclic aryl, cycloalkyl or alkyl group or
`alternatively R‘1 and R‘2 represent —H or form a
`heterocyclic group With the nitrogen atom to Which
`they are attached;
`or B represents a group —OR4 in Which R4 represents an
`aryl, alkyl, polyhydroXyalkyl or cycloalkyl group or a
`group of formula
`
`25
`
`H
`
`Where E represents a group:
`
`R5
`
`R7
`
`in Which R5, R6 and R7 independently represent a hydro
`gen atom or a radical —CHZOH, —CH2O acyl, —OH,
`—O acyl, —NH acyl, —NH2, N+H3X_, X- being
`de?ned an above, or alternatively R5, R6 and R7
`represent a group —COOR8, R8 representing a hydro
`gen atom, a substituted or unsubstituted aralkyl, aryl,
`cycloalkyl or alkyl group, p is a number equal to 1 or
`2 and q is equal to 0 or 1;
`or B represents a halogen atom, or alternatively A repre
`sents a group:
`
`35
`
`45
`
`in Which E is de?ned as above.
`Such ethoXyquine derivatives are described especially in
`Patent Application FR-2,378,796.
`Among the agents capable of neutraliZing singlet oxygen,
`there may be also mentioned polyphenols, that is to say the
`compounds comprising at least one diphenolic aromatic
`ring, it being possible for the phenol groups to be optionally
`etheri?ed or esteri?ed. Among the polyphenols Which can be
`used, there may be mentioned especially the ?avonoids
`corresponding to the general formula (II):
`
`65
`
`in Which A“, B“, C“ and D“, independently of each other,
`represent H or OH; E“ represents H, OR or OX‘, Where
`X‘ represents:
`
`OH
`
`—CO
`
`OH
`
`OH
`
`F“, G“, J“ represent, independently of each other, H or
`OH; and X1 represents —CH2—, —CO— or
`—CHOH—,
`it being understood that at least tWo of the groups A“, B“,
`C“ and D“ or at least tWo of the radicals F“, G“ and J“
`designate a hydroXyl group,
`A‘, C‘ and D‘, independently of each other, represent H,
`OH or OCH3;
`E‘ represents H, or OR‘, Where R‘ represents the residue of
`a sugar of formula R‘OH;
`B‘, F‘, G‘ and J‘, independently of each other, represent H,
`OH, OCH3 or —OCH2—CH2—OH, it being under
`stood that at least tWo of the groups A‘, B‘, C‘ and D‘ do
`not designate —H or that at least one of the groups F‘,
`G‘ and J‘ does not designate —H.
`Among the sugars R‘OH, rutinose may be mentioned.
`The compounds of formula (II) and (III) are knoWn. They
`may be obtained especially according to the procedures
`described in “The Flavonoids” Harborne J. B., Mabry T. J.,
`Helga Mabry, 1975, pages 1 to 45.
`Among the ?avonoids Which can be used according to the
`invention, there may be mentioned especially taXifoline,
`catechin, epicatechin, eriodictyol, naringenin, rutin,
`troXerutin, chrysin, tangeretine, luteolin, opigallocatechin,
`epigallocatechin gallate, quercetin, ?setin, kaempferol,
`galangin, gallocatechin and epicatechin gallate.
`Such compounds occur especially in the green tea eXtracts
`sold under the name Sunphenon by the company Nikko.
`
`3 of 9
`
`

`

`US 6,316,012 B1
`
`5
`Among the polyphonols Which can be used, there may be
`also mentioned polyphenols such as carnosic acid and
`carnosol Which may be extracted for example from rosemary
`either by extraction followed by a distillation (Chang et al.
`JOSC, Vol. 61, No.6, June 1984), or by an extraction With a
`polar solvent such as ethanol preceded by an extraction by
`means of. a non-polar solvent such as hexane in order to
`remove odorous substances, as described in Patent Appli
`cation EP-307 626.
`The polyphenols Which may be used may also be chosen
`from the (2,5-dihydroxyphenyl)alkylcarboxylic acids of for
`mula (IV) and their derivatives (especially esters and
`amides):
`
`20
`
`25
`
`35
`
`45
`
`in Which:
`
`R“1 represents —O—Alc, OH or —N(r‘)(r“), Alc being a
`linear or branched C1—C2O alkyl optionally substituted
`by one or more hydroxyl or alkoxy groups, or Alc being
`a C2—C2O alkenyl,
`r‘ and r“ independently represent H, C1—C2O alkyl, C2—C6
`hydroxyalkyl or C3—C6 polyhydroxyalkyl, or alterna
`tively r‘ and r“ together form, With the nitrogen atom to
`Which they are attached, a heterocycle,
`
`r in a number, including Zero, such that the chain
`—(CH2)r —COR1 comprises at most 21 carbon atoms,
`R“2 and R“3 independently represent H or a C1—C4 alkyl,
`it being possible for R“2 to represent, in addition, a
`C1—C4 alkoxy.
`The compounds of formula (IV) are knoWn or can be
`prepared according to knoWn methods, for example analo
`gous to those described in Patents FR-2,400,358 and FR-2,
`400,359.
`Among the polyphenols Which can be used according to
`the invention, there may be also mentioned the esters or
`amides of caffeic acid. Among the asters of caffeic acid,
`there may be mentioned especially the compounds of for
`mula (V):
`
`(V)
`
`55
`
`HO
`
`HO
`
`in Which Z represents a C1—C8 alkyl, for example methyl,
`or the residue of a phytol.
`Among the amides of caffeic acid, there may be men
`tioned especially the compounds of formula (VI):
`
`65
`
`(VI)
`
`HO
`
`in Which Z‘ represents a C1—C8, in particular C6—C8, alkyl.
`The compounds of formula (V) or (VI) are knoWn or can
`be prepared according to knoWn methods.
`Among the antioxidant. capable of neutraliZing singlet
`oxygen, there may be also mentioned carotenoid derivatives,
`and in particular the folloWing compounds:
`All-trans-betacarotono,
`alpha-carotene,
`gamma-carotene,
`delta-carotene,
`docapreno-beta-carotene,
`dodecaprono-beta-carotene
`lycopen,
`Zaxanthin,
`astaxanthin,
`violaxanthin,
`lutein,
`bixin,
`canthaxnthin,
`cryptoxanthin.
`Among the antioxidants capable of neutraliZing singlet
`oxygen, there may also be mentioned nucleosides and their
`derivatives.
`The nucleosides (for example adenosine, guanosine,
`cytidine, thymidine and uridine and the corresponding deox
`yribose derivatives) are especially those derived from the
`combination of a purine or pyrimidine base chosen from
`adenine, guanine, cytosine, thymine and uracile (abbreviated
`A, G, C, T, U) and a pentose (especially ribose and
`deoxyribose) The nuclooside derivatives are for example
`mono- di- or triphosphates, and especially 3‘- and/or
`5‘-phosphates, as Well an the oligo nucleotides having for
`example up to 20 nucleotide units.
`In the composition of the invention, the proportion by
`Weight of product having a peroxidase activity capable of
`reducing organic peroxides may vary from 0.005% to 5%,
`and in particular from 0.01% to 3%.
`The proportion by Weight of antioxidant capable of neu
`traliZing singlet oxygen may vary for example from 0.005%
`to 3%, and in particular from 0.01% to 1%.
`The relative proportions of peroxidase and anti-singlet
`oxygen may be determined in each case by simple routine
`experiments in Which the relative proportions giving favour
`able results (synergy) are selected, for example in the ODC
`induction test described by R. L. Binder at al., publication
`mentioned above.
`Generally, the peroxidase/anti-singlet oxygen Weight ratio
`may vary for example from 0.001 to 0.3. This ratio is de?ned
`here arbitrarily for a product With a peroxidase activity
`having an activity corresponding to 80 enZymatic units per
`mg. It in therefore easy to adapt this ratio in the came of a
`product With peroxidase activity having a different titre in
`enZymatic units. The peroxidase unit is de?ned beloW in the
`experimental section.
`For the production of the pharmaceutical or cosmetic
`forms, according to knoWn techniques, the solubility char
`
`4 of 9
`
`

`

`US 6,316,012 B1
`
`7
`acteristics of the ingredients Will obviously be taken into
`account, in association With the type of composition desired.
`Products having an organic peroxide-reducing peroxidase
`activity, as Well as nucleosides and their derivatives, are
`generally soluble in hydrophilic, especially aqueous, phases.
`The quinoline derivatives, polyphenols and carotenoids
`are generally soluble in lipophilic phases.
`The compositions of the invention may be provided
`especially in the form of solutions (lotion type
`compositions), thickened solutions, gals, ointments, emul
`sions (creams, milks), vesicular dispersions, poWders, dense
`poWders, pastes or solid sticks. They may also be packaged,
`Where appropriate in pressure-packs containing a propelling
`agent permitting application in the form of foams or sprays.
`The cosmetic or pharmaceutical compositions of the
`invention may contain, in addition to the combination of
`active ingredients described above, the ingredients or adju
`vants customarily used in the production of such
`compositions, and in particular solvents such as Water,
`organic solvents (for example alcohols, oils), or silicones,
`thickening agents, surface-active agents, polymars, solid
`fatty substances (for example Waxes, lanolin), moisturiZing
`agents, preserving agents, pH-modifying agents, sequester
`ing agents, colouring agents, perfumes, solid ?llers
`(poWders and pigments), ultraviolet radiation-absorbing
`substances, self-tanning agents (such an dihydroxyacetone),
`and the like.
`The compositions in the form of vesicular dispersions
`contain for example at least one active ingredient incorpo
`rated into micelles or lipid double layers, Which may encap
`sulate an aqueous phase, and Which are dispersed in an
`aqueous solvent.
`The vesicular dispersions of lipids, especially of ionic or
`non-ionic amphiphilic lipids, are prepared according to
`knoWn processes, for example by sWelling the lipids in an
`aqueous solution in order to form spherules dispersed in the
`aqueous medium, as described in the publication by
`Banghan, Standish and Watkins, J. Mol. Biol. 13,238 (1965)
`or in Patents FR 2,315,991 and 2,416,008 by the Applicant.
`The description of various preparation procedures can also
`be found in “Les liposomes en biologic callulaire et
`pharmacologie”, Inserm/John Libbery Eurotext Edition,
`1987, pages 6 to 18.
`The composition of the invention may contain, in addition
`to the combination described above, other antioxidants such
`as ascorbic acid, magnesium ascorbylphosphate, 0t, [3, y
`and/or o-tocopherols, bilirubin, biliverdine, C1—C1O alkyl
`esters of glutathione, and the like.
`The compositions of the invention are especially cosmetic
`or pharmaceutical compositions Which protect the human
`epidermis, the hair and the mucous membranes, makeup
`compositions for the skin and super?cial body groWths,
`compositions for buccodental use such as dentifrices, or
`ophthalmic compositions such as collyria.
`When the cosmetic composition according to the inven
`tion is used for protecting the hair, it may be provided in the
`form of shampoos, lotions, gals or compositions to be rinsed,
`to be applied before or after shampooing, before or after
`dyeing or bleaching, or before, during or after permanent
`Waving or hair straightening treatment. It may also be
`provided in the form of hair-styling or treating lotions or
`gals, lotions or gals for bloWdrying or hair setting, hair
`lacquers, compositions for permanent Waving or hair
`straightening, or compositions for dyeing or bleaching the
`hair.
`When the composition of the invention is used as makeup
`product for the eyelashes, the eyebroWs or the skin, it is
`
`10
`
`15
`
`25
`
`35
`
`45
`
`55
`
`65
`
`8
`provided for example in the form of creams for treating the
`epidermis, foundations, lipsticks, eyeshadoWs, blushers,
`eyeliners or mascaras.
`The compositions of the invention, and more particularly
`the makeup compositions and the anti-sun compositions
`may contain pigments of metallic oxides such as titanium,
`Zinc, cerium or Zirconium oxides, generally at a concentra
`tion of betWeen 0.1 and 15%, and in particular betWeen 0.5
`and 10% by Weight relative to the total Weight of the
`composition. These pigments are preferably used in the form
`of nanopigments With a mean diameter of loan than 100 nm,
`generally of betWeen 5 and 50 nm. These nanopigments may
`be optionally coated. The coated pigments are pigments
`Which have undergone one or more surface treatments of
`chemical, electronic and/or mechanical nature, With com
`pounds such as amino acids, beesWax, fatty acids, fatty
`alcohols, anionic surface-active agents, lecithins, fatty acid
`salts (salts of sodium, potassium, Zinc, iron or aluminium),
`metallic alkoxides (especially of titanium or aluminium),
`polyethylene, silicones, proteins (for example collagen,
`elastin), alkanolamines, silicon oxides, metallic oxides or
`sodiumhexametaphosphate; see on this subject Cosmetics
`and Toiletries, February 1990, Vol.105, pp.53—64.
`When the composition of the invention is a pharmaceu
`tical composition, it may be provided especially in the form
`of an emulsion (milk or cream), gal, lotion, ointment,
`vesicular dispersion, and may contain, in addition to the
`combination described above, another pharmaceutical active
`ingredient.
`By virtue of the synergistic peroxidase+anti-singlet oxy
`gen combination, the compositions of the invention consti
`tute cosmetic or pharmaceutical compositions intended to be
`applied especially to the skin, super?cial body groWth,
`mucous membranes, Which make it possible especially to
`prevent and treat the damage caused by the free radicals
`induced especially by atmospheric pollutants and by ultra
`violet radiation. In particular, the cosmetic compositions of
`the invention make it possible to prevent or treat especially
`the phenomenon of accelerated aging of the skin. The
`compositions of the invention make it possible, in addition,
`to prevent or limit the risks of skin cancers induced by
`ultraviolet radiation.
`One of the additional advantages of the antioxidant com
`bination according to the invention in that it makes it
`possible to inhibit or decrease the photoinduced reaction
`Which appears When pigments of metallic oxides are
`exposed to light, and Which in detrimental to the stability of
`the compositions, in particular When the latter also contain
`lipids.
`The subject of the invention is also the use, in
`combination, of at least one product With peroxidase activity
`capable of reducing organic peroxides and of at least one
`antioxidant capable of neutraliZing singlet oxygen, as syn
`ergistic active combination in the preparation of a cosmetic
`or pharmaceutical composition intended to prevent or treat
`the cellular damage caused by the free radicals induced
`especially by atmospheric pollutants and/or by ultraviolet
`radiation, and/or intended to combat the phenomenon of
`accelerated aging of the skin, or to prevent or limit the risks
`of photoinduced skin tumours.
`The subject of the invention is also a cosmetic treatment
`process Which makes it possible to combat the aesthetic
`damage caused on the skin and the hair by the free radicals
`induced especially by atmospheric pollutants and by ultra
`violet radiation, characteriZed by the fact that a composition
`containing the synergistic combination Which has been
`described above is applied to the skin or the hair.
`
`5 of 9
`
`

`

`US 6,316,012 B1
`
`The following examples illustrate the invention.
`In these examples, the origin or nature of the ingredients
`is the folloWing:
`Lactoperoxidase (abbreviated LPO): in the form of a
`poWder; obtained from the company Sederma (France).
`Ethoxyquins: mixture of monomer, dimer and polymers
`of 6-ethoxy-1,2-dihydro-2,2,4-trimethylquinoline mar
`keted under the nano Santoquin by the company Mon
`santo
`Guanosines: obtained from Pharma Waldhol
`Compound A: non-ionic amphiphilic compound of for
`mula:
`
`10
`
`10
`0.1% of lactoperoxidase is added thereto, folloWed by the
`phase A beloW:
`
`Phase A
`
`Perfume
`Sunflower oil
`Para?in oil
`Vitamin F
`Soyabean lecithin
`Ascorbyl palmitate
`Hexadecylamine salicylate
`Ethoxyquine
`
`0.4%
`10%
`4%
`2%
`1%
`1%
`0.2%
`0.8%
`
`CHZOH
`
`The mixture is homogeniZed by means of an ultradispers
`ing device, the phase B, consisting of:
`
`20
`
`“Carbopol 940”
`Demineralised Water
`
`0.4%
`79.7%
`
`in then dispersed.
`The Whole is ?nally neutraliZed by means of 0.4% tri
`ethanolamine.
`
`EXAMPLE 3
`
`Beauty Milk for the Body
`
`An oil-in-Water (O/W) emulsion of the folloWing com
`position Was prepared:
`
`Purcellin oil (Dragoco)
`Vaseline oil
`Oleyl alcohol
`Isopropyl myristate
`Glycerin monostearate
`Stearin
`Cetyl alcohol
`Perfume
`Carbopol 941
`Pure triethanolamine
`Butyl para-hydroxybenzoate
`Preserving agent
`Propylene glycol
`Hydroviton
`Guanosine
`Colorant F.D.C. blue 1 (Kohnstamn)
`at 1% in Water
`Lactoperoxidase
`Demineralized Water
`
`2 g
`6 g
`1 g
`1.5 g
`2 g
`1.4 g
`0.1 g
`0.9 g
`0.35 g
`1.05 g
`0.04 g
`0.3 g
`5 g
`1.5 g
`0.5 g
`0.03 g
`
`0.1 g
`71.55 g
`
`EXAMPLE 4
`
`Body Cream
`
`An O/W emulsion of the folloWing composition is pre
`pared:
`
`Cetyl alcohol
`Sipol Wax
`Glycerol monostearate
`Vaseline oil
`Isopropyl myristate
`Glycerin
`Perfume
`
`0.5 g
`5 g
`1.5 g
`6 g
`3 g
`10 g
`0.2 g
`
`in Which R is a hexadecyl radical and n has a mean statistical
`value equal to 3,
`Hydroviton: mixture of amino acids, hydrating ingredi
`ents for the skin and sodium lactate, allantoin (buffer),
`marketed by Dragoco,
`Sipol Wax: partially oxyethylenated cetyl stearyl alcohol,
`marketed by Sinnova-France,
`Carbopols: cross-linked carboxyvinyl polymers marketed
`by Goodrich.
`
`EXAMPLE 1
`
`O/W Emulsion
`
`Lactoperoxidase (LPO)
`Ethoxyquine
`Polyethylene glycol monostearate 50 E0 (ICI)
`Mixture of diglycerol mono- and distearate
`(Dubois Stearin Industries, France)
`Vaseline oil
`Cetyl alcohol
`Water q.s
`
`0.05%
`0.26%
`1.5%
`1.5%
`
`24%
`2.5%
`100%
`
`30
`
`35
`
`40
`
`This emulsion constitutes a cream. To prepare it, the
`aqueous phase in poured into the fatty phase at 80° C., With
`stirring.
`This composition is applied to the face.
`
`45
`
`EXAMPLE 2
`
`Body Care Fluid
`
`Acarrier in the form of a dispersion of lipid spherules, of
`the folloWing composition (in % relative to the ?nal
`composition), is ?rst prepared in a knoWn manner:
`
`Compound A
`Cholesterol
`Dicetyl phosphate
`Methyl para-hydroxybenzoate
`Sterile demineralized Water
`
`4.5%
`4.5%
`1.0%
`0.3%
`30%
`
`For that, the ?rst three ingredients are mixed by melting
`at 100° C., under a nitrogen atmosphere, cooled to 80° C.
`and then homogeniZed by means of a Virtis type ultradis
`persing device. The Water and the preserving agent are then
`added. The dispersion in adjusted to room temperature and
`
`50
`
`55
`
`60
`
`65
`
`6 of 9
`
`

`

`US 6,316,012 B1
`
`-continued
`
`Guanosine
`Lactoperoxidase
`Water q.s.
`
`1.5 g
`0.2 g
`100 g
`
`5
`
`EXAMPLE 5
`
`Body Cream
`An O/W emulsion of the folloWing composition 15 pre
`pared:
`
`.
`
`.
`
`.
`
`.
`
`.
`
`10
`
`Water q.s.
`
`Synperonic PE/L62"
`Propylene glycol
`Ethoxyquine
`Lactic acid
`Tetrasodium salt of
`ethylenediaminetetraacetic acid
`Lactoperoxidase
`Phenoxyethanol
`
`0.2 g
`4 g
`0.1 g
`1 g
`0.1 g
`
`0.4 g
`0.25 g
`
`100 g
`
`sipol WM
`Gwen” monostearate
`Sodium stearate
`Vaseline oil
`Isopropyl palrnitate
`Ethoxyquine
`Glycerin
`Perfume
`Lactoperoxidase
`Water q_s_
`
`6 g
`1'5 g
`0.8 g
`6 g
`2 g
`1 g
`15 g
`0.3 g
`2 g
`100 g
`
`EXAMPLE 6
`Dermopharmaceutical Cream
`
`PhLA
`
`Polyethylene glycol,stearate sold under
`the name Myr] 49 by the company ICI
`Glycerol stearate and polyethylene glycol
`stearate sold under the name “Arlacel 165”
`
`by the company ICI
`Cetyl alcohol
`Stearyl alcohol
`
`Vaseline oil
`
`Steam acid
`Phase B
`
`cross_linked polyacrylic acid S01 d
`under the name “Carbopol 941” by
`the Company Goodrich
`léflfltcilr’iegoxldase
`Methyl para_hydroxybenzoate
`Tirisogiumfalt Of
`_
`_d
`et y en iaminetetraacetic aci
`Triethanolamine (20% aqueous solution)
`W t
`Phase; C
`
`Ethoxyquine
`
`.
`Retinoic acid
`Isopropyl myristate
`
`1.75 g
`
`1.75 g
`
`0.6 g
`0.6 g
`
`17 g
`
`25 g
`
`04 g
`
`0;:
`01 g
`0-1 g
`0.5 g
`66.53
`g
`
`0.05 g
`
`002 g
`5 g
`
`The constituents of the fatty phase A are mixed at 70° C.
`The constituents of the aqueous phase B, heated to 70° C., 55
`are solubiliZed With stirring.
`
`.
`
`.
`
`.
`
`.
`
`.
`
`.
`
`The aqueous phase B is added to the fatty phase A With
`stirring. Ethoxyquine and retinoic acid, solubiliZed in iso-
`propyl myristate (Phase C) are added to the emulsion
`obtained
`An anti-aging treatment cream is obtained Which can be
`applied to the face,
`
`EXAMPLE 7
`Anti-Wrinkle Dermapharmaceutical Gel
`The folloWing compounds are mixed at room temperature
`and With stirring:
`
`*Marketed by the company ICI: block polymer polyoxyethylene/
`polyoxypropylene/polyoxyethylene: Poloxamer 182 (CPTA).
`
`1 g of cross-linked polyacrylic acid sold under the name
`15 Carbopol 940 by the company Goodrich in added to the
`dispersion, and then sodium hydroxide is added in order to
`d. tth Ht 5
`a Jus
`e p
`O '
`The gel obtained in applied to the face and the neck.
`
`20
`
`25
`
`3O
`
`EXAMPLE 8
`
`-
`Protective Care Cream
`
`“SinnoWax A0”
`Glycerol stearate
`Cetyl alcohol
`Jojoba oil
`Linoleic acid
`“MT 100 T” (TiO2)
`Ethoxyquine
`Lactoperoxidase
`Preservatives
`vvater q_s_
`
`5 g
`1 g
`1 g
`6 g
`6 g
`5 g
`1 g
`0.04 g
`q_s_
`100 g
`
`35
`
`~
`~
`~
`~
`'
`The fatty phase is heated to 80° C. Titanium oxide in
`added
`
`The aqueous phase 15 poured, With stirring at 80 C., into
`the fatty phase.
`
`.
`
`.
`
`.
`
`.
`
`o
`
`.
`
`40
`
`This composition, in the form of a cream, in applied to the
`face'
`SinnoWax