`
`Request for Certification
`
`August 31, 2017
`
`
`To: Commissioner, JPO
`
`
`5
`
`1. Indication of Case:
`
`Japanese Patent Application H7-346518
`
`
`
`
`2. Requester:
`
`10
`
`15
`
`
`
`
`
`
`
`
`
`
`
`
`
`Postal Code:
`Address:
`
`100-6620
`Shiga International Patent Office
`
`
`
`
`
`
`
`
`
`
`
`
`
`Name:
`
`
`
`
`
`
`
`
`
`GranTokyo South Tower
`
`1-9-2 Marunouchi, Chiyoda-ku
`
`Tokyo
`
`Yasuhiko MURAYAMA
`
`
`3. Name of Document relating to Certification
`
`20
`
`
`
`
`
`
`Japanese Unexamined Patent Application Publication
`(JP H9-157153 A)
`
`Please certify that the matters disclosed in the “Name
`of Document relating to Certification” are true.
`
`
`
`
`25
`
`
`It is hereby certified that the “Name of Document
`
`relating to Certification” is true.
`
`
`
`
`
`
`
`
`
`
`
`
`
`30
`
`35
`
`September 8, 2017
`
`Naoko MUNAKATA, Commissioner, JPO [SEAL]
`
`
`
`
`
`
`
`
`
` 2017, Certificate No. 600340
`
`L'OREAL USA, INC. EX. 1006
`
`- 1 -
`
`

`

`H9-157153
`
` (12) Laid Open Patent Gazette (A)
`(43) Date Published 17 June 1997
`Code PO Ref. No.
`FI
`
`Technical
`designation
`
`
`A61K 7/48
`
`
`
` 7/00
`
`
`
`
`
`
`
`
`AED
`
`
`
`A61K 7/48
`
`
`
`7/00 F
`
`K
`
`W
`
`31/70 AED
`
`(19) Japanese Patent Office (JP)
`
`- 2 -
`
`(11) Unexamined Patent No.
`
`(51) Int. Cl.6
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`31/70
`
`
`
`
`
`
`
`
`Examination: not yet No. of claims 6 FD (Total 8 pages)
`Continued on the last page
`
`5
`
`(21) Application no.
`
`Application JP H07-346518
`
`(22) Filing date
`
`11 December 1995
`
`(71) Applicant 000135324
`
`
`
`
`
`
`
`
`
`
`10
`
`Noevir Co. Ltd.
`6-13-1
`Minatojimanaka-cho,
`Kobe-shi, Hyogo-ken
`
`Chuo-ku,
`
`(72) Inventor Masumi Takei
`
`
`
`
`
`
`
`Noevir Co. Ltd. Shiga Central Research
`
`Institute, 112-1 Unokami Okada-cho,
`
`Youkaichi-shi, Shiga-ken
`
`15
`
`(74) Agent
`
`Masumi Takei
`
`
`
`
`
`
`
`
`
`
`
`(54) [Title of the invention]
`
`
`
`
`
`20
`
`Preparation for external use on the skin
`
`(57) [Abstract]
`
`
`
`[Problem] To offer preparations for external use
`
`on the skin which have a synergically intensified
`
`activated oxygen species eliminating action, and can
`
`25
`
`prevent skin ageing and injury caused by intra- and
`
`extracorporeal oxidation stress.
`
`
`
`[Means for solving the problem] One or two or more
`
`selected from adenosine and derivatives thereof having
`
`a hydroxy radical eliminating action, and an activated
`
`30
`
`oxygen species eliminating agent selected from
`
`hamamelitannin, extracts of witch hazel, oriental white
`
`

`

`- 3 -
`
`oak, Mongolian oak, horse chestnut, great burnet and
`
`tree peony, thioredoxin, thioredoxin reductase and a
`
`complex thereof and/or placental extract are used
`
`concomitantly, included in a base for an external
`
`5
`
`preparation for use on the skin. Adenosine derivatives
`
`include adenosine monophosphate, adenosine diphosphate,
`
`adenosine
`
`triphosphate
`
`and
`
`cyclic
`
`adenosine
`
`monophosphate.
`
`

`

`- 4 -
`
`
`
`[Claims]
`
`
`
`[Claim 1] Preparation for external use on the
`
`skin, constituted by containing one or two or more
`
`5
`
`selected from adenosine and derivatives thereof, and
`
`hamamelitannin.
`
`
`
`[Claim 2] Preparation for external use on the skin
`
`constituted by containing one or two or more selected
`
`from adenosine and derivatives thereof, and one or two
`
`10
`
`or more extracts of a plant selected from witch hazel,
`
`(Hamamelis japonica Sieb. et Zucc., Hamamelis obtusata
`
`Makino, Hamamelis virginiana L.), oriental white oak
`
`(Quercus serata Thunb.), Mongolian oak (Quercus
`
`crispula
`
`Blume),
`
`horse
`
`chestnut,
`
`(Aesculus
`
`15
`
`hippocastanum
`
`L.),
`
`great
`
`burnet
`
`(Sanguisorba
`
`officinalis L.) and tree peony (Paeonia suffruticosa
`
`Andr.).
`
`
`
`[Claim 3] Preparation for external use on the skin
`
`constituted by containing one or two or more selected
`
`20
`
`from adenosine and derivatives thereof, and one or two
`
`or more of thioredoxin, thioredoxin reductase and a
`
`complex thereof,
`
`
`
`[Claim 4] Preparation for external use on the skin
`
`constituted by containing one or two or more selected
`
`25
`
`from adenosine and derivatives thereof, and one or two
`
`or more of thioredoxin, thioredoxin reductase and a
`
`complex thereof, and reduced nicotinamide adenosine
`
`nucleotide phosphate.
`
`
`
`[Claim 5] Preparation for external use on the skin
`
`30
`
`constituted by containing one or two or more selected
`
`from adenosine and derivatives thereof, and a placental
`
`extract.
`
`
`
`[Claim 6] Preparation for external use on the skin
`
`according to claim 1 to claim 5, characterized in that
`
`

`

`the adenosine derivatives are adenosine monophosphate,
`
`adenosine diphosphate, adenosine triphosphate and
`
`- 5 -
`
`cyclic adenosine monophosphate.
`
`
`
`5
`
`[Detailed Description of the Invention]
`
`[0001]
`
`[Technical field of the invention] The present
`
`invention relates to preparations for external use on
`
`the skin, which have a synergically intensified
`
`10
`
`activated oxygen species eliminating action and can
`
`prevent skin ageing and injury caused by intra- and
`
`extracorporeal oxidation stress. More specifically, it
`
`relates to preparations for external use on the skin
`
`constituted by concomitant use of adenosine and
`
`15
`
`derivatives thereof, and (a) constituent(s) having an
`
`activated oxygen species eliminating action.
`
`[0002]
`
`[Prior art] UV radiation and the internal
`
`metabolism of the body produce as activated oxygen
`
`20
`
`species predominantly hydrogen peroxide, and hydroxy
`
`radicals, singlet oxygen and superoxide, within the
`
`body; it is well known that these have various adverse
`
`effects on the body. In relation in particular to skin
`
`tissue, it has been suggested that such activated
`
`25
`
`oxygen species are profoundly associated with skin
`
`ageing phenomena such as wrinkle formation and
`
`modification of dermal constituents, as well as
`
`peroxidation of skin lipids.
`
`[0003] In order to prevent or improve skin ageing
`
`30
`
`and injury caused by such activated oxygen species,
`
`substances which eliminate these have been screened for
`
`a long time, and vitamin E group compounds, or plant
`
`tannins such as tea tannin, carotenoids, or other
`
`animal or plant extract constituents, etc., have been
`
`35
`
`employed.
`
`

`

`- 6 -
`
`[0004] However, conventionally used activated
`
`oxygen species eliminating agents have poor stability,
`
`and the eliminating action is weak and inadequate,
`
`especially when included in preparations for external
`
`5
`
`use on the skin with a complex formulation, in most
`
`cases a satisfactory effect is not obtained. In
`
`addition, activated oxygen species produced in the body
`
`contribute to complex chain reactions, and if only one
`
`type of activated oxygen species is eliminated it does
`
`10
`
`not necessarily mean that an adequate preventing or
`
`improving action on skin ageing and injury can be
`
`obtained.
`
`[0005]
`
`[Problem which the invention is intended to solve]
`
`15
`
`Therefore, the object of the present invention is to
`
`give preparations for external use on the skin which
`
`synergistically heighten the activated oxygen species
`
`eliminating
`
`action,
`
`effectively
`
`prevent
`
`complex
`
`peroxidation, reactions in the body, and which can
`
`20
`
`effectively prevent and improve skin ageing and injury.
`
`[0006]
`
`[Means for solving the problem] After carrying out
`
`various studies designed to solve the problem above,
`
`the present inventors discovered that by concomitant
`
`25
`
`use of one or two or more selected from adenosine and
`
`derivatives thereof, and a substance or constituent
`
`having a certain activated oxygen species eliminating
`
`action, activated oxygen species eliminating activity
`
`is
`
`stably
`
`not
`
`only
`
`not
`
`lowered
`
`but
`
`rather
`
`30
`
`synergistically heightened, on inclusion in various
`
`bases for preparations for external use on the skin,
`
`and they have perfected the present invention.
`
`[0007] Thus, in the present invention, one or two
`
`or more of adenosine and derivatives thereof, and
`
`35
`
`hamamelitannin, an extract of one or two or more plants
`
`

`

`- 7 -
`
`selected from witch hazel (Hamamelis japonica Sieb. et
`
`Zucc., Hamamelis obtusata Makino, Hamamelis virginiana
`
`L.), oriental white oak (Quercus serata Thunb.),
`
`Mongolian oak (Quercus crispula Blume), horse chestnut,
`
`5
`
`(Aesculus hippocastanum L.), great burnet (Sanguisorba
`
`officinalis L.) and tree peony (Paeonia suffruticosa
`
`Andr.), one or two or more of thioredoxin, thioredoxin
`
`reductase and a complex thereof, or moreover reduced
`
`nicotinamide adenosine nucleotide phosphate and/or
`
`10
`
`placental extract are used concomitantly.
`
`[0008] Adenosine has a hydroxy radical eliminating
`
`action, and has been reported to be efficacious in
`
`suppressing posttraumatic epilepsy (Free Rad. Biol. &
`
`Med.19 (4) 473-479 (1995)). It has now been discovered
`
`15
`
`that the activated oxygen species eliminating action in
`
`biological tissue, and especially skin tissue, is
`
`synergistically heightened by concomitant use of (a)
`
`substance(s) or constituent(s) mentioned above, having
`
`an activated oxygen species eliminating action. The
`
`20
`
`mechanism which gives this synergistic action is
`
`unclear; however, stabilization of the eliminating
`
`action on diverse activated oxygen species, and an
`
`intensification of the eliminating action in the
`
`oxidation chain reactions in the body, etc., can be
`
`25
`
`considered.
`
`[0009] In the case of concomitant use of one or
`
`two or more of adenosine and derivatives thereof, and
`
`(an)
`
`aforementioned
`
`activated
`
`oxygen
`
`species
`
`eliminating agent(s), a protective effect against cell
`
`30
`
`damage due to ultraviolet radiation is shown, as
`
`follows.
`
`[0010] Murine keratinocytes were cultured at 37°C
`
`for 24 hours in Kitano et al. modified MCDB 153 culture
`
`medium, followed by washing twice with phosphate-
`
`35
`
`buffered saline, and then resuspended in Hanks buffer,
`
`

`

`- 8 -
`
`and the different samples shown in Table 1 were
`
`separately added, and then the cells were exposed to
`
`medium wavelength UV radiation (UVB) at 300 mJ/cm2,
`
`with an FL-20S/E lamp as a light source. After
`
`5
`
`exposure, the keratinocytes were incubated at 37ºC for
`
`24 hours in MCDB 153 culture medium; the cell survival
`
`rate was found by neutral red uptake assay, and the
`
`survival rate with addition of each sample was
`
`expressed against the survival rate in the control
`
`10
`
`culture system not exposed to UVB as 100%, and shown in
`
`Table 2
`
`[Table 1]
`
`Activated oxygen
`species
`eliminating
`agents
`Adenosine
`Hamamelitannin
`
`Adenosine
`
`hazel
`
`
`
`
`Sample
`
`1
`
`
`
`2
`
`
`
`3
`
`
`
`4
`
`Final
`concentration
`(µg/ml〕
`
`Sample
`
`250.0
`
`1.0
`
`250.0
`
`5.0
`
`250.0
`
`5.0
`
`250.0
`
`5.0
`
`10
`
`11
`
`12
`
`13
`
`14
`
`
`
`15
`
`16
`
`Activated oxygen
`species
`eliminating
`agents
`Adenosine
`Adenosine
`monophosphate
`Adenosine
`diphosphate
`Adenosine
`triphosphate
`
`Cyclic adenosine
`monophosphate
`
`Hamamelitannin
`
`hazel
`
`Final
`concentration
`(µg/ml〕
`
`250.0
`
`250.0
`
`250.0
`
`250.0
`
`250.0
`
`1.0
`
`5.0
`
`oak
`
`Witch
`extract
`Adenosine
`monophosphate
`Horse
`chestnut
`extract
`Adenosine
`monophosphate
`Oriental
`white
`oak extract
`Adenosine
`diphosphate
`Mongolian
`extract
`Adenosine
`diphosphate
`Great
`burnet
`extract
`Adenosine
`triphosphate
`Tree
`peony
`extract
`Adenosine
`triphosphate
`Thioredoxin-
`thioredoxin
`reductase
`complex
`Cyclic adenosine
`monophosphate
`Placental
`extract
`
`
`
`5
`
`
`
`6
`
`
`
`7
`
`
`
`
`
`8
`
`
`
`
`
`9
`
`
`
`250.0
`
`5.0
`
`250.0
`
`5.0
`
`250.0
`
`2.0
`
`250.0
`
`10.0
`
`250.0
`
`50.0
`
`17
`
`18
`
`19
`
`20
`
`21
`
`22
`
`23
`
`24
`
`
`
`
`
`oak
`
`Witch
`extract
`Horse
`chestnut
`extract
`white
`Oriental
`oak extract
`Mongolian
`extract
`Great
`extract
`Tree
`extract
`Thioredoxin-
`thioredoxin
`reductase
`complex
`
`burnet
`
`peony
`
`Placental
`extract
`Hanks buffer
`
`
`
`
`
`5.0
`
`5.0
`
`5.0
`
`5.0
`
`2.0
`
`10.0
`
`50.0
`
`-
`
`
`
`
`
`15
`
`
`* In the table, the plant extracts were extracted with 50 wt% ethanol, and
`placental extract was extracted with water.
`
`
`
`[Table 2]
`
`
`
`

`

`- 9 -
`
`Cell survival rate (%)
`95.1
`9l.7
`88.0
`81.5
`82.2
`83.4
`92.2
`74.8
`70.3
`58.5
`54.1
`53.9
`
`Sample
`13
`14
`15
`16
`17
`18
`19
`20
`21
`22
`23
`24
`
`Cell survival rate (%)
`52.5
`58.7
`60.4
`65.8
`64.0
`57.3
`55.6
`62.1
`67.5
`54.9
`51.2
`47.2
`
`Sample
`1
`2
`3
`4
`5
`6
`7
`8
`9
`10
`11
`12
`
`
`[0011] From Table 2, a fall in cell survival rate
`
`to 47.2% of that in the control culture system is noted
`
`when exposed to UVB after transfer to Hanks buffer
`
`5
`
`without the addition of an activated oxygen species
`
`eliminating agent (sample 24). In systems with
`
`adenosine or a derivative thereof, or an activated
`
`oxygen species eliminating agent such as hamamelitannin
`
`added thereto to give the final concentrations shown in
`
`10
`
`Table 1 (sample 10 to sample 23), although an elevation
`
`was also noted in cell survival rate, adequate recovery
`
`was not obtained. On the other hand, in systems with
`
`both adenosine or a derivative thereof, and an
`
`activated oxygen species eliminating agent such as
`
`15
`
`hamamelitannin added (sample 1 to sample 9), a
`
`considerable recovery of cell survival rate was noted.
`
`[0012]
`
`[Modes
`
`for
`
`carrying
`
`out
`
`the
`
`invention]
`
`Preparations for external use on the skin according to
`
`20
`
`the present invention can be presented in forms such as
`
`lotions, emulsions, creams and ointments. They can also
`
`

`

`- 10 -
`
`be presented, in the form of skin cosmetics such as
`
`toilet water, beauty serum or lotions. As the inclusion
`
`rates of adenosine or a derivative thereof, and
`
`hamamelitannin, etc., in the external preparation or
`
`5
`
`cosmetic base, about 0.01-10 wt% and 0.0001-10 wt%,
`
`respectively, are suitable, considering the effect of
`
`the base and the effective dose for an action on the
`
`skin, etc. Moreover, additives such as moisturizers,
`
`antioxidants, preservatives, fragrances and UV blocking
`
`10
`
`agents can also be included.
`
`[0013]
`
`[Examples] The features of the present invention
`
`are described in detail by means of practical examples.
`
`[0014] Table 3 shows formulations for skin cream
`
`15
`
`according to the present invention. The "activated
`
`oxygen species eliminating agents" in Table 3 are shown
`
`in Table 4. This skin cream is prepared by mixing (1)-
`
`(5) in Table 3, and then dissolving by heating,
`
`bringing to 75°C, adding thereto (6)-(9) and (12) mixed
`
`20
`
`and dissolved by heating and brought to 75°C,
`
`emulsification, cooling with stirring; and then mixing
`
`in (10) and (11) at 40°C.
`
`[Table 3]
`
`
`
`Constituent
`
`
`1 Stearic acid
`2 Cetanol
`3 Glycerol monostearate
`4 Squalane
`5 Sorbitan monooleate
`
`Inclusion rate (wt%)
`1.00
`0.50
`0.50
`20.00
`2.00
`
`

`

`- 11 -
`
`6
`
`Poly(ethylene oxide)(20EO) sorbitan
`monostearate
`7 Sodium hydroxide
`8 Carboxyvinyl polymer
`9 Methyl paraoxybenzoate
`Activated oxygen species eliminating
`agent
`11 Fragrances
`12 Pure water
`
`Total quantity
`
`10
`
`
`
`
`
`[Table 4]
`
`Example
`
`Activated oxygen species eliminating agent
`
`1
`
`
`2
`
`3
`
`4
`
`
`5
`
`6
`
`7
`
`
`
`
`Adenosine
`Hamamelitannin
`Adenosine monophosphate
`Witch hazel 50 wt% ethanol extract
`Adenosine diphosphate
`Horse chestnut 50 wt% ethanol extract
`Adenosine triphosphate
`Oriental white oak ethanol extract
`Great burnet ethanol extract
`Cyclic adenosine monophosphate
`Tree peony 1,3-butylene glycol extract
`Adenosine
`Thioredoxin-thioredoxin reductase complex
`Reduced nicotinamide adenosine nucleotide phosphate
`Adenosine monophosphate
`Placenta aqueous extract
`
`2.00
`
`0.05
`0.10
`0.10
`
`0.20
`
`0.20
`73.35
`100.00
`
`Inclusion rate
`(wt%)
`0.10
`0.10
`0.10
`0.10
`0.10
`0.10
`0.10
`0.05
`0.05
`0.10
`0.10
`0.05
`0.10
`0.05
`0.10
`0.10
`
`[0015] The aforementioned examples of the present
`
`5
`
`invention were studied for the effect in preventing
`
`skin ageing, by evaluating prevention of skin
`
`wrinkling. When hairless mice are exposed to long
`
`wavelength UV radiation (UVA), wrinkle development is
`
`promoted, and the effect in preventing this development
`
`10
`
`of wrinkling due to UVA was evaluated. Hairless mice
`
`were put into groups of 5; in each group, an example of
`
`the present invention and a comparative example were
`
`separately applied once daily to the back, which was
`
`exposed for 50 weeks to UVA at 1 J/cm2/week, and the
`
`15
`
`prevalence of wrinkling at this time was evaluated
`
`visually. It should be noted that as comparative
`
`examples, creams in which the activated oxygen species
`
`eliminating agents in Table 3 were replaced with the
`
`activated oxygen species eliminating agents shown in
`
`20
`
`Table 5 were used. The prevalence of wrinkling was
`
`scored with "no noticeable wrinkle development 0",
`
`

`

`- 12 -
`
`"slightly noticeable minute wrinkle development 1",
`
`"clearly noticeable slight wrinkle development 2",
`
`"noticeable development of moderate wrinkling 3",
`
`"noticeable development of deep wrinkling 4", the
`
`5
`
`average scores for each group were calculated and the
`
`relationship with number of days of exposure to UVA is
`
`shown in Table 6.
`
`[Table 5]
`
`
`
`Compara
`tive
`example
`1
`2
`3
`4
`5
`6
`7
`8
`
`9
`10
`11
`
`12
`
`Activated oxygen species eliminating agent
`
`Inclusion rate
`(wt%)
`
`Adenosine
`Hamamelitannin
`Adenosine monophosphate
`Witch hazel 50 wt% ethanol extract
`Adenosine diphosphate
`Horse chestnut 50 wt% ethanol extract
`Adenosine triphosphate
`Oriental white oak ethanol extract
`Great burnet ethanol extract
`Cyclic adenosine monophosphate
`Tree peony 1,3-butylene glycol extract
`Thioredoxin-thioredoxin reductase complex
`Reduced nicotinamide adenosine nucleotide phosphate
`Placenta aqueous extract
`
`0.20
`0.20
`0.20
`0.20
`0.20
`0.20
`0.20
`0.10
`0.10
`0.20
`0.20
`0.15
`0.05
`0.20
`
`10
`
`
`
`
`
`
`
`
`[Table 6]
`
`Days of exposure to UVA (weeks)>
`0
`20
`40
`50
`0.2
`0.6
`1.8
`3.8
`0.2
`0.2
`0.4
`0.6
`0
`0.2
`0.4
`0.6
`0.4
`0.4
`0.6
`0.8
`0.2
`0.2
`0.6
`1.0
`0.2
`0.2
`0.4
`0.6
`0.2
`0.2
`0.6
`1.2
`0.4
`0.4
`0.8
`1.2
`0.4
`0.6
`1.0
`1.8
`0.2
`0.2
`0.8
`1.4
`0.2
`0.4
`1.2
`2.0
`0.2
`0.4
`1.0
`1.6
`0.4
`0.6
`1.0
`2.0
`0.2
`0.4
`0.6
`1.6
`0.2
`0.4
`1.0
`2.0
`0.2
`0.4
`0.8
`1.8
`0
`0.4
`1.0
`2.2
`0.2
`0.6
`0.8
`1.8
`0.4
`0.6
`1.0
`2.4
`0.2
`0.6
`1.2
`2.6
`
`
`
`
`Sample
`Control
`1
`2
`3
`4
`5
`6
`7
`1
`2
`3
`4
`5
`6
`7
`8
`9
`10
`11
`12
`
`Examples
`
`Comparative examples
`
`
`
`

`

`- 13 -
`
`[0016] From Table 6, in the control group after 40
`
`weeks of UVA exposure wrinkle development was clearly
`
`noticeable, and after 50 weeks deep wrinkle development
`
`was noted in nearly all mice. By contrast, a marked
`
`5
`
`effect in preventing development of wrinkling was seen
`
`in the groups to which the examples of the present
`
`invention were applied, with only minute wrinkle
`
`development being noted after 50 weeks. On the other
`
`hand, in the groups coated with the comparative
`
`10
`
`examples which contained only adenosine or a derivative
`
`thereof, or hamamelitannin, etc., as an activated
`
`oxygen species eliminating agent, although an effect
`
`was noted in preventing the development of wrinkling,
`
`it was very small compared with the groups coated with
`
`15
`
`an example of the invention, wrinkle development was
`
`clearly noticeable in nearly all mice, and, especially
`
`in the group coated with comparative example 12,
`
`wrinkling reached almost a moderate depth.
`
`[0017] Moreover, other formulations of examples of
`
`20
`
`the present invention are presented below.
`
`[0018]
`
`[Example 8] Oil-in-water type ointment
`
`
`
`(1) liquid paraffin
`
`10.0 (wt%)
`
`(2) white petrolatum jelly
`
`(3) cetanol
`
`(4) polyoxyethylene (30EO) cetyl ether
`
`5.0
`
`6.0
`
`2.0
`
`(5) autoemulsifying glycerol monostearate
`
`10.0
`
`(6) propylene glycol
`
`(7) methyl paraoxybenzoate
`
`(8) adenosine
`
`(9) adenosine triphosphate
`
`(10) hamamelitannin
`
`(11) pure water
`
`10.0
`
`0.1
`
`0.1
`
`0.1
`
`0.1
`
`56.6
`
`Preparation Method: (1)-(5) are mixed, dissolved
`
`25
`
`by heating, and held at 75°C. On the other hand, (6),
`
`

`

`- 14 -
`
`(7) and (11) are mixed, dissolved, and heated to 75°C;
`
`the aforementioned oil phase constituents are added
`
`gradually thereto with stirring and emulsified; after
`
`cooling, (8)-(10) are added at 40°C.
`
`5
`
`[0019]
`[Example 9] Lotion
`
`(1) polyoxyethylene (20EO) sorbitan
`
` 1.00 (wt%)
`
`monolaurate
`
`(2) glycerol
`(3) propylene glycol
`(4) ethanol
`(5) methyl paraoxybenzoate
`(6) adenosine monophosphate
`(7) adenosine diphosphate
`(8) cyclic adenosine monophosphate
`(9) witch hazel ethanol extract
`(10) great burnet ethanol extract
`(11) pure water
`
`Preparation Method: All the constituents (1)-(10)
`are added in this order to (11), followed by mixing and
`homogenization.
`
`10.00
` 5.00
`20.00
` 0.10
` 0.02
` 0.02
` 0.02
` 0.05
` 0.05
`63.74
`
`10
`
`[0020]
`[Example 10] Toilet water
`
`(1) decaglyceryl monolaurate
`(2) 1, 3-butylene glycol
`(3) sorbitol
`(4) ethanol
`(5) methyl paraoxybenzoate
`(6) adenosine monophosphate
`(7) cyclic adenosine monophosphate
`(8) tree peony ethanol extract
`(9) fragrances
`(10) pure water
`
`
` 1.00 (wt%)
` 3.00
` 2.00
` 2.00
` 0.10
` 0.02
` 0.01
` 0.01
` 0.20
`91.66
`
`15
`
`Preparation Method: All the constituents (1)-(9)
`
`are added in this order to (10), followed by mixing and
`
`homogenization.
`
`[0021]
`
`[Example 11] Skin lotion
`
`(1) squalane
`
`(2) white petrolatum jelly
`
` 5.00 (wt%)
`
` 2.00
`
`

`

`- 15 -
`
`(3) beeswax
`
`(4) sorbitan sesquioleate
`
` 0.50
`
` 0.80
`
`(5) polyoxyethylene (20EO) oleyl ether
`
` 1.20
`
`(6) propylene glycol
`
` 5.00
`
`(7) carboxyvinyl polymer 1.0 wt% aqueous
`
`20.00
`
`solution
`
`(8) methyl paraoxybenzoate
`
`(9) potassium hydroxide
`
`(10) adenosine
`
`(11) adenosine monophosphate
`
`(12) horse chestnut propylene glycol
`
`extract
`
`(13) great burnet glycerol extract
`
`(14) fragrances
`
`(15) pure water
`
`
`
` 0.10
`
` 0.10
`
` 0.02
`
` 0.02
`
` 0.01
`
`0.01
`
`0.10
`
`65.14
`
`Preparation Method: The oil phase constituents
`
`(1)-(5) are mixed, dissolved and homogenized, and
`
`heated to 75°C. On the other hand, the aqueous phase
`
`5
`
`constituents (6), (8) and (15) are mixed, dissolved and
`
`heated to 75°C; the aforementioned oil phase
`
`constituents are added thereto, followed by preliminary
`
`emulsification, and then (7) is added, followed by
`
`uniform emulsification in a homomixer. After cooling,
`
`10
`
`(9) is added and the pH is adjusted, and (10)-(14) are
`
`mixed in at 40°C.
`
`[0022]
`[Example 12] Skin cream
`
`(1) beeswax
`
`(2) cetanol
`
`(3) reduced lanolin
`
`(4) squalane
`
`(5) glyceryl fatty acid ester
`
`(6) lipophilic glycerol monostearate
`
`(7) polyoxyethylene (20EO) sorbitan
`
`monolaurate
`
`(8) propylene glycol
`
` 6.00 (wt%)
`
` 5.00
`
` 8.00
`
`37.50
`
` 4.00
`
` 2.00
`
` 2.00
`
` 5.00
`
`

`

`- 16 -
`
`(9) methyl paraoxybenzoate
`
`(10) adenosine
`
` 0.10
`
` 0.02
`
`(11) witch hazel 50 wt% ethanol extract
`
` 0.01
`
`(12) thioredoxin-thioredoxin reductase
`
` 0.10
`
`complex
`
`(13) fragrances
`
`(14) pure water
`
`
`
` 0.15
`
`30.12
`
`Preparation Method: The oil phase constituents
`
`(1)-(7) are mixed, dissolved and heated to 75°C. On the
`
`other hand, the aqueous phase constituents (8), (9) and
`
`5
`
`(14) are mixed, dissolved and heated to 75°C, the
`
`aforementioned oil phase is added thereto, followed by
`
`emulsification, and after cooling, (10)-(13) are added
`
`at 40°C.
`
`[0023]
`
`10
`
`[Example 13] Skin cream
`
`(1) stearic acid
`
`(2) stearyl alcohol
`
`(3) glycerol monostearate
`
`(4) hardened castor oil
`
`(5) liquid paraffin
`
`(6) jojoba oil
`
`(7) sorbitan sesquioleate
`
`(8) carboxyvinyl polymer
`
`(9) sodium hydroxide
`
`(10) methyl paraoxybenzoate
`
`(11) adenosine triphosphate
`
` 1.00 (wt%)
`
` 4.00
`
` 3.00
`
` 7.00
`
`10.00
`
` 2.00
`
` 1.00
`
` 0.10
`
` 0.05
`
` 0.10
`
` 0.05
`
`(12) tree peony 50 wt% ethanol extract 0.02
`
`(13) placenta aqueous extract
`
`(14) fragrances
`
`(15) pure water
`
`
`
` 0.20
`
` 0.10
`
`71.38
`
`

`

`- 17 -
`
`Preparation Method: (1)-(7) are mixed, dissolved
`
`by heating, and held at 75°C. On the other hand, (8)-
`
`(10) and (15) are mixed, dissolved and heated to 75°C,
`
`the aforementioned oil phase is added thereto followed
`
`5
`
`by emulsification, and cooling, and (11)-(14) are added
`
`at 40°C.
`
`[0024] Usage tests were carried out on the
`
`aforementioned example 8 to example 13 of the present
`
`invention. In the usage tests, there were groups of 20
`
`10
`
`panellists ordinarily employed for field tests, the
`
`different groups used the examples and comparative
`
`examples separately in a blinded fashion on the face
`
`and hands, and evaluation was carried out by observing
`
`changes in wrinkling and skin elasticity. The period of
`
`15
`
`use was 1 year, from April to March. Wrinkling was
`
`evaluated by 5 stages "decrease", "slight decrease",
`
`"no change", "minute increase in wrinkling", and "clear
`
`increase in wrinkling", skin elasticity was evaluated
`
`by 5 stages "raised", "slightly raised", "no change",
`
`20
`
`"slight decrease", and "decrease", each evaluation is
`
`presented in Table 7 as the numbers of panellists
`
`giving each score.
`
`[0025] It should be noted that as comparative
`
`examples, the formulation of example 8 without the
`
`25
`
`inclusion of adenosine, adenosine triphosphate and
`
`hamamelitannin
`
`was
`
`comparative
`
`example
`
`13,
`
`a
`
`formulation without the inclusion of hamamelitannin was
`
`comparative example 14, the formulation of example 9
`
`without the inclusion of adenosine monophosphate,
`
`30
`
`adenosine diphosphate and cyclic adenosine mono-
`
`phosphate was comparative example 15, the formulation
`
`of example 10 without the inclusion of tree peony
`
`extract was comparative example 16, the formulation of
`
`example 11 without the inclusion of adenosine and
`
`35
`
`adenosine monophosphate was comparative example 17, the
`
`

`

`- 18 -
`
`formulation of example 12 without the inclusion of
`
`witch
`
`hazel
`
`extract
`
`and
`
`Thioredoxin-thioredoxin
`
`reductase complex was comparative example 18, and the
`
`formulation of example 13 without the inclusion of
`
`5
`
`adenosine triphosphate was comparative example 19,
`
`these were all made up to 100 wt% with pure water.
`
`[0026]
`
`[Table 7]
`
`Sample
`Evaluation
`Decrease
`Slight decrease
`No change
`Minute increase
`Clear increase
`Raised
`Slightly raised
`No change
`Slight decrease
`Decrease
`
`9
`8
`13 10
`7
`8
`0
`2
`0
`0
`0
`0
`15 12
`5
`7
`0
`1
`0
`0
`0
`0
`
`Comparative examples
`Examples
`10 11 12 13 13 14 15 16 17 18 19
`4
`6
`2
`2
`0
`0
`0
`0
`0
`0
`0
`5
`6
`3
`2
`0
`3
`4
`3
`2
`1
`3
`11
`8
`15 16
`2
`12 11 11 10 10 13
`0
`0
`0
`0
`14
`5
`5
`6
`8
`8
`4
`0
`0
`0
`0
`4
`0
`0
`0
`0
`1
`0
`5
`8
`5
`4
`0
`0
`0
`0
`0
`0
`0
`9
`10
`5
`5
`0
`5
`5
`3
`4
`0
`2
`6
`2
`10 11
`0
`12 11
`9
`10
`8
`10
`0
`0
`0
`0
`7
`3
`4
`7
`5
`9
`7
`0
`0
`0
`0
`13 0
`0
`1
`1
`3
`1
`
`Item
`
`ng
`
`Wrinkli
`
`ity
`
`elastic
`Skin
`
`
`
`10
`
`In Table 7 a tendency towards an increase in
`
`wrinkling and a clear decrease in skin elasticity is
`
`noted in the group using comparative example 13 which
`
`did not include an activated oxygen species eliminating
`
`agent. By contrast, no panellists noted an increase in
`
`15
`
`wrinkling and a decrease in skin elasticity in the
`
`groups using the examples of the present invention, and
`
`in particular, in the group using example 8, all of the
`
`panellists noted a decrease in wrinkling and a tendency
`
`towards raised skin elasticity, and in the group using
`
`20
`
`example 9, 90% of panellists noted a decrease in
`
`wrinkling, and 95% of panellists noted a tendency
`
`towards raised skin elasticity.
`
`[0027] On the other hand, in the groups using
`
`Comparative example 14 to Comparative example 19,
`
`25
`
`containing only one or two or more selected from
`
`adenosine and derivatives thereof or only an activated
`
`oxygen species eliminating agent such as hamameli-
`
`tannin, although an effect in preventing an increase in
`
`wrinkling and a decrease in skin elasticity is noted
`
`

`

`- 19 -
`
`compared with the group using comparative example 13,
`
`they do not show a marked improving tendency relative
`
`to this group.
`
`[0028] It should be noted that no panellists noted
`
`5
`
`skin irritation or sensitization in the groups using
`
`the examples of the present invention during the period
`
`of the usage test.
`
`[0029]
`
`[Effects of the invention] As discussed in detail
`
`10
`
`above, with the present invention it is possible to
`
`offer preparations for external use on the skin which
`
`provide a synergistic increase in activated oxygen
`
`species eliminating action, and can prevent and
`
`moreover effectively improve skin ageing and injury
`
`15
`
`caused by intra- and extracorporeal oxidation stress.
`
`
`
`Continued from the front page
`
`Code PO Ref. No.
`
`FI
`
`
`
`
`(51) Int. Cl.6
`
` A61K 35/50
`
`
`
`35/78
`
`
`
`38/44
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`ADA
`ADS
`
`Technical
`designation
`
`
`A61K 35/50
`
`
`
`35/78 F
`
`W
`
`ADAC
`
`37/50 ADS
`
`
`
`
`

`

`-20-
`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`
`I, Roland Piers LEVAN BA (Joint Hons),
`
`translator to RWS Group Ltd, of Europa House, Chiltern Park, Chiltern Hill, Chalfont St
`
`Peter, Buckinghamshire, United Kingdom,declare;
`
`Li
`
`2.
`
`3
`
`That I am a citizen of the United Kingdom of Great Britain and Northern Ireland.
`
`That I am well acquainted with the Japanese and English languages.
`
`That the attached is, to the best of my knowledge andbelief, a true translation into the
`
`English language of JPH09157153.
`
`4.
`
`That | believe that all statements made herein of my own knowledge are true and that
`
`all statements made on information and belief are true; and further that these statements were
`
`made with the knowledgethat willful false statements and the like so made are punishable by
`
`fine or imprisonment, or both, under Section 1001 of Title 18 of the United States Code and
`
`that such willful false statements may jeopardize the validity of the patent application in the
`
`United States of America or any patent issuing thereon.
`
`Date: February 28, 2018
`
`R. P. LEVAN
`
`For and on behalf of RWS Group Ltd
`
`- 20 -
`
`