U.S. patent application number 11/259882 was filed with the patent office on 2006-02-16 for method for measuring skin absorbability.
This patent application is currently assigned to SHISEIDO COMPANY., LTD.. Invention is credited to Ryuichi Takamoto, Eriko Takeoka, Toshio Yanaki.
Application Number | 20060034762 11/259882 |
Document ID | / |
Family ID | 27339025 |
Filed Date | 2006-02-16 |
United States Patent
Application |
20060034762 |
Kind Code |
A1 |
Takeoka; Eriko ; et
al. |
February 16, 2006 |
Method for measuring skin absorbability
Abstract
A method for measuring skin absorbability of a substance, which
includes measuring the amount of the substance which is penetrated
into hair follicles; and using the resultant data as an index
representing the amount of the substance which is absorbed through
skin pores, to thereby determine the amount of the substance
absorbed through skin pores and a method for measuring skin
absorbability of a substance, which includes bringing the substance
into contact with a first surface of a thin film that mimics the
skin surface layer, a second surface of the film being brought into
contact with artificial sebum; and evaluating the sebum
transferability of the substance by using the degree of transfer of
the substance to the artificial sebum as an index, are disclosed. A
kit for performing the measurement method of the present invention
is also disclosed.
Inventors: |
Takeoka; Eriko; (Kanagawa,
JP) ; Takamoto; Ryuichi; (Tokyo, JP) ; Yanaki;
Toshio; (Kanagawa, JP) |
Correspondence
Address: |
Law Office of Townsend & Banta;c/o PortfolioIP
P.O. Box 52050
Minneapolis
MN
55402
US
|
Assignee: |
SHISEIDO COMPANY., LTD.
|
Family ID: |
27339025 |
Appl. No.: |
11/259882 |
Filed: |
October 27, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
10238520 |
Sep 9, 2002 |
|
|
|
11259882 |
Oct 27, 2005 |
|
|
|
Current U.S.
Class: |
424/9.1 ;
435/4 |
Current CPC
Class: |
A61Q 5/02 20130101; A61Q
5/00 20130101; A61Q 7/00 20130101; A61K 8/342 20130101; A61Q 9/00
20130101; G01N 33/5088 20130101 |
Class at
Publication: |
424/009.1 ;
435/004 |
International
Class: |
A61K 49/00 20060101
A61K049/00; C12Q 1/00 20060101 C12Q001/00 |
Foreign Application Data
Date |
Code |
Application Number |
Oct 29, 1999 |
JP |
11-309225 |
Dec 17, 1999 |
JP |
11-358840 |
Jun 15, 2000 |
JP |
2000-180457 |
Claims
1-27. (canceled)
28. A method for measuring skin absorbability of a substance which
is delivered to hair follicles comprising the steps of: (a)
applying the substance to a skin model specimen comprising dermis,
said skin model assuming a pseudo-biological state; (b) removing
hair follicles from the dermis of the skin model; (c) measuring the
amount of substance absorbed in the hair follicles; and (d) using
the amount of the substance in the hair follicles as an index
representing the amount of the substance which is absorbed through
skin pores, to thereby determine the amount of the substance
absorbed through skin pores.
29. The method for measuring skin absorbability of a substance
which is delivered to hair follicles according to claim 28,
comprising: removing the hair follicles and other skin tissue in
the skin model; measuring the amount of the substance in the hair
follicles and the amount of the substance in the other skin tissue,
and using the amount of the substance in the hair follicles as an
index representing the amount of the substance which is absorbed
through skin pores, and the amount of the substance in the other
skin tissue as an index representing the amount of the substance
which is absorbed percutaneously, to thereby determine the amount
of the substance absorbed through skin pores.
30. The method for measuring skin absorbability of a substance
which is delivered to hair follicles according to claim 28, wherein
the absorption through skin pores is absorption through hair
follicles.
31. The method for measuring skin absorbability of a substance
which is delivered to hair follicles according to claim 29, wherein
the other skin tissue is stratum corneum, epidermis or dermis.
32. The method for measuring skin absorbability of a substance
which is delivered to hair follicles according to claim 28, wherein
the substance is determined to have skin-pore absorbability.
33. The method for measuring skin absorbability of a substance
which is delivered to hair follicles according to claim 28, further
comprising: applying a second substance, which accelerates
skin-pore absorption of the substance, to the skin model
specimen.
34. The method for measuring skin absorbability of a substance
which is delivered to hair follicles according to claim 28, wherein
the substance is a hair-growing ingredient.
35. A method for measuring skin absorbability of a substance which
is delivered to hair follicles, comprising: (a) applying the
substance to human scalp having hair follicles; (b) removing the
hair follicles from the human scalp; (c) measuring the amount of
substance in the hair follicles; and (d) using the amount of the
substance in the hair follicles as an index representing the amount
of the substance which is absorbed through skin pores, to thereby
determine the amount, of the substance absorbed through skin
pores.
36. The method for measuring skin absorbability of a substance
which is delivered to hair follicles according to claim 35, wherein
the absorption through skin pores is absorption through hair
follicles.
37. The method for measuring skin absorbability of a substance
which is delivered to hair follicles according to claim 35, wherein
the substance is determined to have skin-pore absorbability.
38. The method for measuring skin absorbability of a substance
which is delivered to hair follicles according to claim 35, further
comprising: applying a second substance, which accelerates
skin-pore absorption of the substance, to the skin model
specimen.
39. The method for measuring skin absorbability of a substance
which is delivered to hair follicles according to claim 35, wherein
the substance is a hair-growing ingredient.
40. A method for measuring skin absorbability of a substance which
transfers to sebum, comprising the steps of: (a) bringing the
substance into contact with a first surface of a thin film which
mimics the skin surface layer, a second surface of the film being
brought into contact with artificial sebum; (b) evaluating the
degree of transfer of the substance to the artificial sebum through
the thin film, and (c) evaluating the sebum transferability of the
substance by using the degree of transfer of the substance to the
artificial sebum as an index.
41. The method for measuring skin absorbability of a substance
which transfers to sebum according to claim 40, wherein the first
surface of the film comprises an open system.
42. The method for measuring skin absorbability of a substance
which transfers to sebum according to claim 40, wherein the
artificial sebum assumes a fluid state.
43. The method for measuring skin absorbability of a substance
which transfers to sebum according to claim 40, wherein the thin
film is a single-layer silicone film, a surface of the film having
undergone keratinization, and the substance is brought into contact
with the surface which has undergone keratinization.
44. The method for measuring skin absorbability of a substance
which transfers to sebum according to claim 40, wherein the
substance is determined to have sebum transferability.
45. The method for measuring skin absorbability of a substance
which transfers to sebum according to claim 40, further comprising:
bringing into contact a second substance, which accelerates sebum
transfer of the substance, with the first surface.
46. The method for measuring skin absorbability of a substance
which transfers to sebum according to claim 40, wherein the
substance is a hair-growing ingredient.
47. A kit for carrying out a method for measuring skin
absorbability of a substance as recited in claim 40, comprising: a
sealable receptacle in which artificial sebum may be placed, the
receptacle comprising an opening which includes a fixing means
enabling fixation of a thin film so as to cover the opening in a
state in which the thin film is removable; a thin film which mimics
the skin surface layer, which film is to be applied and fixed onto
the opening of the sealable receptacle so as to cover the opening;
and artificial sebum which is to be placed in the sealable
receptacle.
Description
BACKGROUND OF THE INVENTION
[0001] 1. Field of the Invention
[0002] The present invention relates to a method for screening an
ingredient which is to be applied to the hair; a kit for carrying
out the screening; and a composition comprising an ingredient
exhibiting effects to accelerate absorption of an active ingredient
through skin pores (hereinafter the absorption will be referred to
as "skin-pore absorption") and transfer of the active ingredient to
sebum, the effects being confirmed by means of the screening
method.
[0003] 2. Background Art
[0004] An important step in the development of a hair-growing agent
is to confirm that an active ingredient exhibits excellent
hair-growing effect and has penetrability so as to reliably reach a
target site. Whether or not the active ingredient reliably reaches
the target site greatly depends on the properties of the active
ingredient or the base ingredient of the hair-growing agent.
[0005] The mechanism of absorption of a drug or an active agent
through hair follicles has become of interest, since hair follicles
are considered to be an effective route through which a
hair-growing agent as well as a vaccine or a gene can be
administered.
[0006] The degree of transfer of a drug to sebum (hereinafter the
degree will be referred to as "sebum transferability") has been
reported to influence whether the drug easily reaches a target site
in the skin (e.g., hair follicles or sebaceous gland) (Critical
Reviews, 14 (3): 207 219 (1997)).
[0007] The penetrability of a drug through the hair follicles has
been studied, for example, by the following methods: [0008] (1) a
method for observing tissue sections by means of fluorescence
labeling or radioisotopes (Suzuki, M., et al., J. Soc. Cosmet.
Chem. (1978), Nicholau, G., et al., Xenobiolica (1987), Lieb, L.
M., et al., J. Invest. Dermatol. (1995)) and [0009] (2) a method
for comparing hairy skin and hairless skin with respect to the
transdermal permeability of a drug (e.g. comparison of hairless rat
and hairy rat: Illel, B., et al., J. Pherm. Sci. (1991); comparison
of normal human skin and skin which has received burns and has
subsequently healed: Hueber, F., et al., J. skin Pharmacol.
(1994)).
[0010] However, method (1) is qualitative rather than quantitative,
and the method involves the time-consuming preparation of tissue
sections. In method (2), absorption of a drug is studied only from
the viewpoint of transdermal absorption, and "skin-pore absorption"
and "sebum transferability," which are important properties of a
hair-growing agent, cannot be confirmed directly.
[0011] Therefore, the aforementioned conventional methods encounter
difficulty in directly confirming that a drug reaches a target site
in the skin and exerts the intended effects.
SUMMARY OF THE INVENTION
[0012] In view of the foregoing, an object of the present invention
is to provide means for conveniently and reliably determining the
degree of skin-pore absorption (hereinafter the degree will be
referred to as "skin-pore absorbability") and sebum transferability
of a substance such as a drug, the skin-pore absorbability and
sebum transferability being indexes for directly confirming that
the substance reaches a target site in the skin and exerts the
desired effects. Another object of the present invention is to
provide a composition for the scalp and hair which exhibits
excellent skin-pore absorbability and sebum transferability and
which enables a drug contained in the composition to reliably reach
a target site in the skin, the skin-pore absorbability and the
sebum transferability being determined through the means of the
present invention.
[0013] The sebum transferability of a drug is considered a useful
index to evaluate whether or not the drug easily reaches a specific
target site in the skin, but can not be easily evaluated in vivo.
Therefore, a specific object of the present invention is to provide
means for accurately and conveniently evaluating the sebum
transferability of a drug in vitro.
[0014] The present inventors have performed extensive studies, and
have found that, when the amount of a substance which is penetrated
into hair follicles, the amount being determined in vitro or in
vivo, is employed as an index of the amount of the substance which
is absorbed through skin pores, the skin-pore absorbability of the
substance can be conveniently and reliably determined, to thereby
measure the skin absorbability of the substance.
[0015] As used herein, the term "skin-pore absorption" refers to
absorption through skin pores, although the term "transdermal
absorption" refers to absorption through any portion of the skin
(i.e., over the entire skin area). Therefore, as described below, a
drug which exhibits transdermal absorbability does not necessarily
exhibit skin-pore absorbability. In addition, because a
hair-growing agent exerts its actions in the vicinity of the hair
follicle, evaluating the "skin-pore absorbability" of the agent is
very important.
[0016] The present invention provides a method for the measurement
of skin absorbability of a substance (hereinafter the method will
be referred to as "measurement method 1"), which comprises
measuring the amount of a substance which is penetrated into the
hair follicles; and using the resultant data as an index
representing the amount of the substance which is absorbed through
the skin pores, to thereby determine the skin-pore absorbability of
the substance.
[0017] If the phenomenon of "skin-pore absorption" is considered
from the function of absorption, penetration of a substance into
hair follicles substantially occurs through skin pores. Therefore,
in the present invention, "absorption through skin pores" or
"skin-pore absorption" and "absorption through hair follicles" or
"transfollicular absorption" are used as terms having the
substantially same meaning.
[0018] Accordingly, measurement method 1 is also provided as a
method for the measurement of skin absorbability of a substance,
which comprises measuring the amount of a substance which is
penetrated into the hair follicles; and using the resultant data as
an index representing the amount of the substance which is absorbed
through the hair follicles, to thereby determine the
transfollicular absorbability of the substance.
[0019] Measurement method 1 is classified into the following two
methods: [0020] (1) an in vitro quantification method (hereinafter
referred to as "measurement method 1-1") in which the amount of a
substance (e.g., a drug) which is penetrated into the hair follicle
or skin tissue is determined by applying the substance to a model
skin having a pseudo-biological state, removing the hair follicle
from the dermis or skin tissue of the skin model, and measuring the
concentration of substance which is delivered to the hair follicle
or skin tissue; and [0021] (2) an in vivo quantification method
(hereinafter referred to as "measurement method 1-2") in which the
amount of a substance (e.g., a drug) which is penetrated into the
hair follicle is determined by applying the substance to the human
scalp, removing the hair follicle from the scalp, and measuring the
concentration of the substance which is delivered to the hair
follicle.
[0022] The present inventors have also found that the sebum
transferability of a substance can be easily evaluated in vitro by
use of a skin model comprising artificial sebum.
[0023] The present invention also provides a method for the
measurement of skin absorbability of a substance (hereinafter the
method will be referred to as "measurement method 2"), which
comprises bringing a substance into contact with a first surface of
a film that mimics the skin surface layer, a second surface of the
film being brought into contact with artificial sebum; and using
the degree of transfer of the substance to the artificial sebum as
an index, to thereby evaluate the sebum transferability of the
substance.
[0024] The present inventors have also found that, by means of
measurement methods 1 and 2, an oily ingredient having an I.O.B.
value of 0.06-4.0 exhibits excellent effects of accelerating
skin-pore absorption and sebum transfer of extensive polar
substances from water-soluble substances to fat-soluble
substances.
[0025] The present invention also provides a composition for the
scalp and hair comprising an oily ingredient having an I.O.B. value
of 0.06-4.0 (hereinafter the composition may be referred to as "the
composition of the present invention").
[0026] As used herein, I.O.B. (Inorganic/Organic Balance) value
refers to an index representing the polarity of an oily ingredient
(hereinafter the term "oily ingredient" includes fats and oils,
waxes, hydrocarbon oils, higher fatty acids, higher alcohols,
esters, general alcohols, and fatty acids). The I.O.B. value
represents the ratio of an inorganic value of the oily ingredient
to an organic value of the oily ingredient (note: the ratio is
calculated as follows: an organic value of 20 is assigned for each
carbon atom in the molecule of the oily ingredient, and an
inorganic value of 100 is assigned per hydroxyl group in the
molecule of the oily ingredient, and these values are used as
yardsticks for calculating an inorganic value for another
substituent (inorganic group); see (1) "Organic Analysis" authored
by Fujita (1930), published by Kaniya Shoten, (2). "Prediction of
Organic Compounds and Organic Conceptual Diagram ("Kagaku-no-Ryoiki
11-10" (1957), pp. 719-725, authored by Fujita, (3) "Systematic
Organic Qualitative Analysis (Book of Purified Substances)" (1970),
p 487, authored by Fujita and Akatsuka, published by Kazama Shoten,
(4) "Organic Conceptual Diagram, Its Fundamentals and Applications"
(1984), p 227, authored by Koda, published by Sankyo Shuppan, (5)
"Design of Emulsion Formulations by use of Organic Conceptual
Diagram" (1985), p 98, authored by Yaguchi, published by Nippon
Emulsion K.K., and (6) R. H. Ewell, J. M. Harrison, L. Berg.: Ind
Eng. Chem. 36, 871 (1944)), and is expressed by: I.O.B.=inorganic
value of the oily ingredient/organic value of the oily
ingredient.
[0027] A typical product form of the composition of the present
invention; i.e., the composition which is to be applied to the
scalp and hair, is a hair-growing agent, but the composition is not
particularly limited to the product form. For example, the
composition may assume a variety of product forms, including
compositions for treating hair, such as a hair-removing agent, a
hair-dyeing agent, and a hair-bleaching agent; compositions for
washing hair, such as shampoo and rinse-in-shampoo; compositions
for protecting hair, such as a rinse and a treatment agent;
gray-hair prevention agents; cosmetics which are absorbed through
the skin excluding the scalp; and drugs such as vaccines and
genetic drugs.
[0028] According to the present invention, there is provided means
for conveniently and accurately measuring the skin absorbability of
a test substance through quantification of the skin-pore
absorbability or sebum transferability of the test substance. By
means of the method, there is provided means for accelerating the
skin-pore absorption or sebum transfer of a drug or an active
ingredient, for example in a hair-growing agent, by which the drug
or active ingredient effectively reach the target site; i.e., the
hair follicle.
BRIEF DESCRIPTION OF THE DRAWINGS
[0029] The foregoing and other objects, features, and many of the
attendant advantages of the present invention will be readily
appreciated as the same becomes better understood with reference to
the following detailed description of the preferred embodiments
when considered in connection with accompanying drawings, in
which:
[0030] FIG. 1A and FIG. 1B are schematic representations showing an
embodiment of the kit of the present invention;
[0031] FIG. 2 is a schematic representation to elucidate
measurement method 1-1;
[0032] FIG. 3 shows the results of the evaluation test of the
accuracy of measurement method 1-1;
[0033] FIG. 4 shows the results of the ingredient evaluation by
means of measurement method 1-1;
[0034] FIGS. 5A to 5C show the results of a test in which the
concentrations of pantothenyl ethylether delivered to the stratum
corneum (FIG. 5A), the epidermis (FIG. 5B), and the dermis (FIG.
5C) of a skin sample are measured by means of measurement method
1-1;
[0035] FIG. 6 shows the results of the evaluation of the effects of
oily ingredients having I.O.B. values of 0.06-4.0 for accelerating
the skin-pore absorption of pantothenyl ethylether;
[0036] FIGS. 7A to 7D show the results of evaluation of the effects
of other oily ingredients having I.O.B. values of 0.06-4.0 for
accelerating the skin-pore absorption of pantothenyl
ethylether;
[0037] FIG. 8 shows the results of the evaluation test conducted by
means of measurement method 1-2;
[0038] FIGS. 9A and 9B show the results of a comparison between
isostearyl alcohol and a transdermal absorption-accelerating
agent;
[0039] FIG. 10 shows the results of the evaluation of refreshing
agents conducted by means of measurement method 1-2;
[0040] FIG. 11A and FIG. 11B show the results of the evaluation of
refreshing agents by means of measurement method 1-1; and
[0041] FIG. 12 shows the results of the test in which measurement
method 2 is carried out by use of the kit of the present
invention.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0042] Embodiments of the present invention will next be
described.
[0043] A Measurement Method 1
[0044] Measurement Method 1-1
[0045] Measurement method 1-1 is an embodiment of measurement
method 1 carried out in vitro.
[0046] In measurement method 1-1, the skin of a model animal or the
skin reconstructed from skin cells may be employed as a skin model.
The model animal from which the skin is taken is not particularly
limited, and pig, human, cattle, mouse, or rat are all potential
donors. In measurement method 1-1, the skin of a hairy animal is
usually employed in order to evaluate the penetrability of a drug
through skin pores into hair follicles. Therefore, a mutated
hairless animal is usually unsuitable for the model animal. In
addition, in view that measurement method 1-1 is employed for the
screening of a drug which is to be ultimately applied to humans,
the skin of a model animal which is employed in the method
preferably mimics that of a human. Therefore, other than human, pig
is preferably employed as a model animal.
[0047] In measurement method 1-1, the phrase "the skin has a
pseudo-biological state" refers to the case in which the skin
specimen is highly analogous to the skin of a living organism. The
skin is not particularly limited, so long as the skin satisfies the
above condition. For example, as described below in the Example, a
skin model may be fixed onto a collagen gel.
[0048] Thus, a substance such as a drug is applied to a skin model
specimen having a pseudo-biological state, the hair follicles are
removed from the dermis of the skin model specimen, and the amount
of the substance which is delivered to the hair follicle is
quantified. The delivery amount may be employed as an index of the
skin-pore absorbability of the substance.
[0049] The conditions for the application of the substance may be
arbitrarily determined in accordance with the type of the substance
or the object. Usually, the skin to which the substance is applied
is incubated at about 37.degree. C. (near the temperature of the
human body) over a predetermined time period. The incubation time
varies with the intended product form of the substance.
[0050] The portion at which the delivery amount of the substance is
measured is limited to the region corresponding to the dermis of
the skin, since in the region corresponding to the stratum corneum
or epidermis of the skin, the hair follicle may be contaminated
with epidermal tissue debris or excretions from the sebaceous
glands, and thus the exact delivery amount of the substance is
difficult to measure.
[0051] The method for measuring the delivery amount of the
substance is not particularly limited. For example, liquid
chromatography (including high-performance liquid chromatography),
a fluorescent antibody technique using a fluorescence-labeled
antibody, mass spectrometry, or a method using a radioisotope may
be employed. Of these, liquid chromatography is preferable, in
consideration of convenience.
[0052] When the amount of the substance which is delivered to the
hair follicle is large, the amount being measured through the
aforementioned method, the substance is considered to have
excellent skin-pore absorbability. Incidentally, conventional
qualitative measurement methods provide limited information about
the substance; i.e., whether or not the substance has skin-pore
absorbability.
[0053] The skin absorbability of a substance such as a drug can be
also quantified as follows: the amount of the substance penetrated
into hair follicles and the amount of the substance penetrated into
different portions of the skin tissue, such as the stratum corneum,
epidermis or dermis are determined and the resultant data is used
as an index of the amount of the substance which is absorbed
through hair follicles and percutaneously.
[0054] Thus, the skin absorbability of a substance is measured by
means of measurement method 1-1.
[0055] Measurement Method 1-2
[0056] In measurement method 1-2, a method for the application of a
substance such as a drug to the human scalp may be arbitrarily
determined in accordance with the characteristics or the intended
product form of the substance.
[0057] After the substance is applied to a person participating in
the test, several hair strands from the person are removed, and the
thus-obtained hair follicles may be stored as hair shafts and/or
outer root sheaths. The amount of the substance which is delivered
to the subcutaneous portion of the hair shaft and/or the outer root
sheath is measured, and the thus-obtained delivery amount of the
substance may be employed as an index of the skin-pore
absorbability of the substance.
[0058] The delivery amount of the substance may be measured in a
manner similar to that of measurement method 1-1.
[0059] Thus, the skin absorbability of the substance is measured
through measurement method 1-2.
[0060] In measurement method 1-2, evaluation of a substance
requires removal of human hair. Therefore, when a variety of
substances are to be evaluated for skin-pore absorbability,
measurement method 1-2 is not necessarily appropriate. In order to
evaluate the skin-pore absorbability of the substances, it is
preferable that firstly, they are screened through measurement
method 1-1, and of the resultant screened substances, those which
exhibit relatively high skin-pore absorbability are further
evaluated through measurement method 1-2.
[0061] B. Measurement Method 2
[0062] Artificial sebum which is employed in measurement method 2
is a mixture of oils and fats, and the mixture can be prepared on
the basis of the composition of natural sebum.
[0063] Examples of oily ingredients which are incorporated in the
artificial sebum include, but are not limited to, triglyceride,
C14-C18 saturated fatty acids, C14-C18 unsaturated fatty acids,
squalene, squalane, cholesterol, and C18-C72 wax esters.
[0064] A specific example formulation of the artificial sebum will
be described in the Example.
[0065] In order to accurately evaluate the transferability of a
substance, the artificial sebum preferably assumes a fluid state.
The fluid state is easily formed by stirring of the artificial
sebum by use of, for example, a magnetic stirrer.
[0066] When a surface of a thin film which mimics the real skin not
containing the dermis (hereinafter the film may be simply referred
to "thin film") is brought into contact with the artificial sebum,
a structure similar to that of the skin surface may be
approximated. In this case, a structure similar to that of the
surface layer of the skin is reproduced, since the aperture of the
sebaceous gland (a sebum supply source) is present in the surface
layer. Therefore, a surface of the thin film to which a substance
such as a drug is applied is preferably analogous to the surface
layer of the skin, in order to reproduce the state in which the
substance is applied to the skin. Specifically, a substrate film
may be formed of a thin film which exhibits no surface reactivity,
such as a single-layer silicone film, a polyethylene carbonate
film, a cellulose film, or another polymer film. A surface of the
substrate film is subjected to keratinization or powder treatment,
in order to form a structure similar to that of the stratum
corneum. The resultant treated side of the thin film is regarded as
the stratum corneum (hereinafter the side may be referred to as
"stratum corneum side), to which a substance is applied in the
present invention.
[0067] The sebum transferability of a substance may be evaluated
through the following procedure: the substance is applied to the
stratum corneum side of the thin film, the opposite side of the
stratum corneum side being brought into contact with the artificial
sebum; the degree of transfer of the substance to the artificial
sebum, for example, the amount of the substance transferred to the
sebum per unit time, i.e., the amount of the substance contained in
the sebum is measured; and the amount of the substance in the sebum
is employed as an index, to thereby evaluate the sebum
transferability of the substance.
[0068] The method for measuring the amount of the substance
contained in the artificial sebum is not particularly limited, and
a customary method may be arbitrarily employed in accordance with
the characteristics of the substance. For example, high-performance
liquid chromatography, thin-layer chromatography, or mass
spectrometry may be employed.
[0069] In measurement method 2, in order to reproduce the case in
which a drug is actually applied to the skin, the stratum corneum
side of the thin film is preferably made open to the air. However,
the stratum corneum side may be closed in accordance with the
intended use of a drug (for example, when the penetration
acceleration effect of the drug is intended to be effected in a
closed state).
[0070] Measurement method 2 is specifically carried out through the
following preferred embodiment.
[0071] The present invention provides a kit for carrying out
measurement method 2 (hereinafter the kit may be referred to as
"the kit of the present invention") comprising: [0072] (1) a
sealable receptacle in which artificial sebum is placed, the
receptacle comprising an opening which includes a fixing means
enabling fixation of a thin film so as to cover the opening in the
state in which the thin film is removable; [0073] (2) a thin film
which mimics the skin surface layer, which film is to be applied
and fixed onto the opening of the sealable receptacle so as to
cover the opening; and [0074] (3) artificial sebum which is to be
placed in the sealable receptacle.
[0075] The kit of the present invention will be described with
reference to FIGS. 1A and 1B. FIGS. 1A and 1B are schematic
representations showing an embodiment of the kit of the present
invention.
[0076] With reference to FIG. 1A, a sealable receptacle 11 is a
cylindrical container, one side of the container being an opening
111 and the other serving as the bottom. A protuberant portion of
the opening 111 protrudes towards the outside and around the
circumference of the receptacle 11.
[0077] The sealable receptacle 11 is preferably formed from a
transparent material in order to permit easy observation of the
inside of the receptacle. To enable easy removal of the artificial
sebum from the sealable receptacle, the receptacle is preferably
formed from a material which does not cause leakage of the contents
even when a needle such as an injection needle is inserted through
the receptacle wall.
[0078] The thin film 12 exhibits no surface reactivity, and a first
surface 121 of the film is subjected to keratinization so as to
form a state analogous to the skin surface layer.
[0079] When the kit is used, the thin film 12 is fixed onto the
opening 111 of the receptacle 11 so as to cover the opening
(usually, the first surface 121 of the film is directed towards the
outside of the receptacle 11, and a second surface 122 of the film
is directed towards the inside of the receptacle 11). The sealable
receptacle 11 must be provided with the entirety or a part of a
fixing means which enables fixing of the thin film on the opening,
so that the thin film can be fixed on or removed from the opening.
In the present embodiment, a thin-film fixing member 13 is employed
as the primary fixing means.
[0080] The thin-film fixing member 13 has two openings (openings
131 and 132) formed at opposite ends. The protuberant portion of
the opening 131 protrudes towards the outside and around the
circumference of the member, and the protuberant portion can be
engaged with the corresponding protuberant portion of the opening
111 of the receptacle 11. The structure of these protuberant
portions is not particularly limited, so long as they can engage
with each other. For example, these portions may have a
concave-convex structure so as to enable coupling of the portions.
In addition, arbitrary means may be employed in order to enhance
the degree of engagement or sealing between the sealable receptacle
11 and the member 13.
[0081] As shown in FIG. 1A, a packing 14 is provided between the
first surface 121 of the thin film 12 and the thin-film fixing
member 13, to thereby form the conjunction structure "thin-film
fixing member 13--packing 14--thin film 12--sealable receptacle
11." As a result, the degree of engagement or sealing between the
sealable receptacle 11 and the member 13 can be enhanced. In
general, the packing 14 may be formed from rubber such as silicone
rubber, synthetic rubber, or natural rubber. Of these, silicone
rubber is preferable, in consideration of corrosion resistance.
[0082] In the kit of the present invention, in order to fix the
conjunction structure "thin-film fixing member 13--packing 14--thin
film 12--sealable receptacle 11," a fixing member 15 as shown in
FIG. 1B may be employed. The fixing member 15 comprises two plate
members (152 and 153) and four supporting bars (151a, 151b, 151c,
151d), the length (.PHI.) of the bars being slightly longer than
the length (.phi.) of the conjunction portion of the conjunction
structure "thin-film fixing member 13--packing 14--thin film
12--sealable receptacle 11." The plate members are maintained to be
parallel to each other through support of the bars. The fixing
member 15 may comprise a structure which enables adjustment of the
length (.PHI.) of the bars. For example, a screw structure which
enables adjustment of the length (.PHI.) may be provided with the
supporting bars (151a, 151b, 151c, and 151d), to thereby adjust the
length (.PHI.) in accordance with the length (.phi.). The plate
members 152 and 153 have a concave portion so that the fixing
member 15 can be fitted to the openings 111 and 113. The size of
the concave portion is larger than that of the circumferential
portion of the openings 111 and 131.
[0083] When the conjunction portion of the conjunction structure
"thin-film fixing member 13--packing 14--thin film 12--sealable
receptacle 11" is fitted with the concave portion of the fixing
member 15 and then the conjunction portion is placed within the
space of the fixing member 15, the space being defined by the
supporting bars, the conjunction portion can be supported by the
supporting force of the bars (151a, 151b, 151c, and 151d) between
the plate members 152 and 153. The conjunction portion may be fixed
by use of a clip instead of the fixing member 15 having the
aforementioned structure.
[0084] FIG. 1B shows an embodiment of use of the kit 10. The thin
film 12 is sandwiched, via the packing 14, by the sealable
receptacle 11 and the thin-film fixing member 13, to thereby form
the conjunction structure "thin-film fixing member 13--packing
14--thin film 12--sealable receptacle 11." The thin film 12 is
fixed onto the opening 111 of the sealable receptacle 11 in which
the artificial sebum is placed, so as to cover the opening. The
surface 121 which has undergone keratinization is directed to the
upper side, and the surface 122 is brought into contact with the
artificial sebum in the sealable receptacle 11. A magnetic stirrer
16 is placed on the bottom of the sealable receptacle 11, and thus
the artificial sebum can assume a fluid state. The conjunction
portion may be fixed by means of the fixing member 15.
[0085] A substance is brought into contact with the surface 121 of
the thin film 12 through the opening 132 of the thin-film fixing
member 13. The sebum transferability of the substance is evaluated
by determining the degree of transfer of the substance, via the
thin film 12, to the artificial sebum in the sealable receptacle
11. When evaluating the sebum transferability of the substance is
to be carried out in a closed system, the opening 132 may be closed
by use of a cap.
[0086] The degree of transfer of the substance to the artificial
sebum can be determined by analysis of the sebum after sampling by
use of a syringe 17.
[0087] Thus, the skin absorbability of the substance can be
measured by means of measurement method 2.
[0088] B. Composition of the Present Invention
[0089] As described below in the Example, an oily ingredient having
an I.O.B. value of 0.06-4.0 exhibits effects which accelerate the
skin-pore absorption or sebum transfer of a drug, the effects being
confirmed by means of measurement method 1 or 2.
[0090] Briefly, when one or more oily ingredients having an I.O.B.
value of 0.06-4.0 are incorporated into a composition for the scalp
and hair which may assume the product form of a hair-growing agent,
penetration of a hair-growing ingredient through skin pores into
hair follicles can be accelerated, the hair follicles being the
target of the agent. When the oily ingredient has an I.O.B. of less
than 0.06 or in excess of 4.0, the oily ingredient tends not to
exhibit the desired effects of accelerating the skin-pore
absorption or sebum transfer of the agent.
[0091] Specifically, an oily ingredient having an I.O.B. value of
0.06-4.0 preferably has an organic value of 100-750 and an
inorganic value of 50-400. More preferably, the oily ingredient has
an inorganic value of 100-400.
[0092] Preferably, an oily ingredient having an I.O.B. value of
0.06-4.0 is a hydrocarbon derivative which is liquid at ambient
temperature and has a backbone structure of a C6-C18 hydrocarbon.
The hydrocarbon derivative is preferably an amphipathic substance
which is liquid at ambient temperature. When the hydrocarbon
derivative is liquid at ambient temperature, the number of carbon
atoms of the derivative is usually 24 or less. Examples of the
hydrocarbon derivative include, but are not limited to, fatty
acids, alcohols, amides, and esters. Of these, alcohols or fatty
acids are preferable.
[0093] Specific examples of the aforementioned oily ingredient
include lauryl alcohol (I.O.B. value 0.41: organic value 240,
inorganic value 100), lauric acid (I.O.B. value 0.68: organic value
220, inorganic value 150), lauric acid ethanolamide (I.O.B. value
1.25: organic value 240, inorganic value 300), lauric acid
diethanolamide (I.O.B. value 1.53: organic value 260, inorganic
value 400), monoglyceryl laurate (I.O.B. value 0.92: organic value
280, inorganic value 260), diglyceryl laurate (I.O.B. value 0.32:
organic value 500, inorganic value 160), oleyl alcohol (I.O.B.
value 0.28: organic value 360, inorganic value 102), oleic acid
(I.O.B. value 0.45: organic value 340, inorganic value 152), oleic
acid ethanolamide (I.O.B. value 0.83: organic value 360, inorganic
value 302), oleic acid diethanolamide (I.O.B. value 1.05: organic
value 380, inorganic value 402), monoglyceryl oleate (I.O.B. value
0.66: organic value 400, inorganic value 262), diglyceryl oleate
(I.O.B. value 0.30: organic value 740, inorganic value 222),
isostearyl alcohol (I.O.B. value 0.29: organic value 350, inorganic
value 100), isostearic acid (I.O.B. value 0.45: organic value 330,
inorganic value 150), isostearic acid ethanolamide (I.O.B. value
0.86: organic value 350, inorganic value 300), isostearic acid
diethanolamide (I.O.B. value 1.08: organic value 370, inorganic
value 400), monoglyceryl isostearate (I.O.B. value 0.66: organic
value 390, inorganic value 260), diglyceryl isostearate (I.O.B.
value 0.30: organic value 730, inorganic value 220), stearyl
alcohol (I.O.B. value 0.27: organic value 360, inorganic value
100), stearic acid (I.O.B. value 0.44: organic value 340, inorganic
value 150), stearic acid ethanolamide (I.O.B. value 0.83: organic
value 360, inorganic value 300), stearic acid diethanolamide
(I.O.B. value 1.05: organic value 380, inorganic value 400),
monoglyceryl stearate (I.O.B. value 0.67: organic value 390,
inorganic value 260), diglyceryl stearate (I.O.B. value 0.30:
organic value 740, inorganic value 220), and benzyl alcohol (I.O.B.
value 0.96: organic value 120, inorganic value 115). Of these,
lauryl alcohol, lauric acid, isostearic acid, isostearyl alcohol,
monoglyceryl isostearate, or isostearic acid ethanolamide are
preferable, and isostearyl alcohol is more preferable.
[0094] Terpenes do not necessarily satisfy the above condition;
i.e., I.O.B. value of 0.06-4.0, but they tend to exhibit excellent
effects to accelerate the skin-pore absorption of a drug or an
active ingredient. Particularly, camphor, among terpenes, exhibits
the excellent effect of accelerating skin-pore absorption.
[0095] The composition of the present invention comprises an oily
ingredient having an I.O.B. value of 0.06-4.0, such as isostearyl
alcohol, as an ingredient which accelerates skin-pore absorption of
an active ingredient of a drug or transfer of the active ingredient
to sebum (the composition may comprise a terpene such as camphor
serving as an ingredient which accelerates skin-pore absorption of
the active ingredient, singly or in combination with the oily
ingredient having an I.O.B. value of 0.06-4.0). The oily ingredient
or a terpene may be incorporated into the composition so as to
attain another object. For example, isostearyl alcohol may serve as
an oily agent, a solvent for a drug, or a sebum-dissolving agent. A
terpene such as camphor may serve as a refreshing agent or a
blood-circulation-stimulating agent.
[0096] The amount of the oily ingredient having an I.O.B. value of
0.06-4.0 in the composition of the present invention is not
particularly limited, since the amount of the ingredient varies in
accordance with the product form or use of the composition.
[0097] When the composition assumes the product form of a
hair-growing agent and when the oily ingredient is intended to
exhibit effects for accelerating the skin-pore absorption or sebum
transfer of an active ingredient of the agent, the oily ingredient
is incorporated into the composition in an amount of 1 ppm-20% by
weight of the composition, preferably 0.01-5% by weight of the
composition.
[0098] The composition of the present invention may contain, in
addition to the oily ingredient having an I.O.B. value of 0.06-4.0,
pharmaceutically active ingredients or base ingredients in
accordance with a specific product form of the composition.
[0099] For example, when the composition assumes the product form
of a hair-growing agent, the composition may contain the following
pharmaceutically active ingredients: blood-circulation-stimulating
agents such as Swertia herb extract, cepharanthine, vitamin E and
it derivatives, y-oryzanol, and raspberry extract; topical
stimulation agents such as capsicum tincture, ginger tincture,
cantharides tincture, and benzyl nicotinate; nutrition agents such
as vitamins and amino acids; female hormones such as estradiol and
ethinylestradiol; hair-root activators such as pantothenic acid and
its derivatives, placental extract, allantoin, and photosensitive
pigment 301; and anti-inflammatory agents such as glycyrrhetic acid
and glycyrrhizinic acid.
[0100] The oily ingredient having an I.O.B. value of 0.06-4.0 which
is incorporated into the composition as an essential ingredient
accelerates the skin-pore absorption or sebum transfer of the
pharmaceutically active ingredient, and enhances the penetrability
of the active ingredient into the hair follicle.
[0101] The composition of the present invention may further contain
any known ingredient. Examples of known ingredients include
humectants, corneocyte descuamating agents, refreshing agents, oily
ingredients, surfactants, silicones, ultraviolet absorbents,
perfumes, and water.
EXAMPLES
[0102] The present invention will next be described in more detail
by way of Examples, which should not be construed as limiting the
invention thereto. Unless otherwise specified, the incorporation
amount of an ingredient is represented by wt. % of the
composition.
[0103] Measurement Method 1-1
[0104] 1. Evaluation of Accuracy of Measurement Method 1-1
[0105] In the evaluation test, 0.5% pantothenyl ethylether (90%
ethanol product) was employed as a substance to be tested, and the
skin of a miniature pig (Yucatan Micropig Skin Set: product of
Charles River) was employed as a skin model.
[0106] As shown in FIG. 2, a skin sample 10 from the miniature pig
(size: 2 cm.times.2 cm) (the sample including a stratum corneum 11,
epidermis 12, and dermis 13) was prepared, and the sample was fixed
onto a collagen gel, to thereby assume a pseudo-biological state.
The test sample (20 .mu.l) was applied onto the skin sample
comprising skin pores 14. The substance was applied onto a region
(diameter: 1.5 cm) of the sample; i.e., within an assay ring to
which silicon grease was applied. The resultant skin sample 10 was
allowed to stand at 37.degree. C. for 16 hours. After having being
washed, the sample 10 was separated into the stratum corneum 11,
the epidermis 12, and the dermis 13. Subsequently, hair follicles
15 were removed from the dermis 13, and the amount of pantothenyl
ethylether contained in the follicles was measured, as a delivery
amount per unit weight of the hair follicle, by means of liquid
chromatography (NANOSPACE (product of Shiseido Co., Ltd.), eluent:
(CH.sub.3CN:H.sub.2O=3:7), detection wavelength: 210 nm).
[0107] The skin sample 10 of the miniature pig was derived from the
upper back and lower back (n=3).
[0108] The results of the test are shown in FIG. 3. In FIG. 3, the
vertical axis corresponds to the concentration of pantothenyl
ethylether delivered to the follicles per unit weight of the hair
follicle. As is apparent from FIG. 3, the portion from which the
samples were prepared exhibits little influence on data. The
results show that data of high reproducibility can be obtained by
means of measurement method 1-1.
[0109] 2 Evaluation of Ingredient by Means of Measurement Method
1-1
[0110] (1) Ingredient Evaluation Test 1
[0111] Test samples; i.e., a control composition (ethanol product)
and a 5% isostearyl alcohol-containing composition, were prepared.
Formulations of the respective compositions are shown in Table 1.
TABLE-US-00001 TABLE 1 5% Isostearyl Control alcohol Ingredient
(wt. %) composition composition Ethanol 89.5 84.2 Polyoxyethylene
-- 0.1 hydrogenated castor oil (40 E.O.) Isostearyl alcohol -- 5.0
Isostearic acid -- 0.2 Ion-exchange water 10.0 10.0 Pantothenyl
ethylether 0.5 0.5
[0112] The amount of each of the test samples (the control
composition and the 5% isostearyl alcohol-containing composition)
which was absorbed through skin pores was measured in a manner
similar to that of the above accuracy test. Skin samples were
derived from the upper back and lower back of the miniature pig
(n=5). The results of the evaluation test are shown in FIG. 4. As
is apparent from FIG. 4, the concentration of pantothenyl
ethylether delivered to hair follicles increases significantly in
the presence of isostearyl alcohol.
[0113] In the test, the concentration of pantothenyl ethylether
delivered to the stratum corneum, the epidermis, and the dermis of
each skin sample was measured by means of liquid chromatography as
described above. The results are shown in FIG. 5A, FIG. 5B, and
FIG. 5C.
[0114] As is apparent from FIGS. 5A-5C, the concentration of
pantothenyl ethylether delivered to the dermis is slightly
increased in the presence of isostearyl alcohol. In contrast, the
concentration of pantothenyl ethylether delivered to the stratum
corneum or the epidermis is decreased in the presence of isostearyl
alcohol. The results show that isostearyl alcohol specifically
accelerates the skin-pore absorption of pantothenyl ethylether, but
does not accelerate the transdermal absorption thereof.
[0115] In the test, it is shown that "skin-pore absorption" or
"transfollicular absortption" is an index of skin absorption
different from "transdermal absorption", and in addition, that it
is possible to measure skin absorption of a substance by regarding
the result shown in FIG. 4 as a concentration of the substance
delivered to hair follicles and regarding the results shown in
FIGS. 5A-5C as concentrations of the substance delivered to other
skin tissue of the skin of the miniature pig (stratum corneum,
epidermis and dermis), and then using the resultant data as an
index representing the amount of the substance which is absorbed
through hair follicles and percutaneously.
[0116] (2) Ingredient Evaluation Test 2
[0117] Test samples comprising an oily ingredient having an I.O.B.
value of 0.06-4.0 were prepared. Formulations of the respective
samples are shown in Table 2. TABLE-US-00002 TABLE 2 1% Isostearyl
Control alcohol Ingredient (wt. %) composition composition
Pantothenyl ethylether 2.0 2.0 99% Ethanol 88.0 87.0 Ion-exchange
water 10.0 10.0 Isostearyl alcohol -- 1.0 Benzyl alcohol -- --
Isostearic acid -- -- Lauric acid -- -- diethanolamide Monoglyceryl
isostearate -- -- 1% Benzyl alcohol 1% Isostearic Ingredient (wt.
%) composition acid composition Pantothenyl ethylether 2.0 2.0 99%
Ethanol 87.0 87.0 Ion-exchange water 10.0 10.0 Isostearyl alcohol
-- -- Benzyl alcohol 1.0 -- Isostearic acid -- 1.0 Lauric acid --
-- diethanolamide Monoglyceryl isostearate -- -- 1% Lauric acid 1%
monoglyceryl diethanolamide isostearate Ingredient (wt. %)
composition composition Pantothenyl ethylether 2.0 2.0 99% Ethanol
87.0 87.0 Ion-exchange water 10.0 10.0 Isostearyl alcohol -- --
Benzyl alcohol -- -- Isostearic acid -- -- Lauric acid 1.0 --
diethanolamide Monoglyceryl isostearate -- 1.0
[0118] In each of six test samples, the concentration of
pantothenyl ethylether which was delivered to the hair follicles
was measured in a manner similar to that of the above accuracy
test. Skin samples were derived from the upper back and lower back
of the miniature pig (n=5). The results of the evaluation test are
shown in FIG. 6. As is apparent from FIG. 6, in comparison with the
control composition, in the compositions containing the oily
ingredient having an I.O.B value of 0.06-4.0, the concentration of
pantothenyl ethylether delivered to hair follicles increases.
Particularly, in the isostearyl alcohol-containing composition and
isostearic acid-containing composition, the concentration of
pantothenyl ethylether delivered to hair follicles increases.
[0119] (3) Ingredient Evaluation Test 3
[0120] Test samples comprising an oily ingredient having an I.O.B.
value of 0.06-4.0 were prepared. The oily ingredient was chosen
from among lauryl alcohol, lauric acid, lauric acid ethanolamide,
lauric acid diethanolamide, monoglyceryl laurate, diglyceryl
laurate, oleyl alcohol, oleic acid, oleic acid ethanolamide, oleic
acid diethanolamide, monoglyceryl oleate, diglyceryl oleate,
isostearyl alcohol, isostearic acid, isostearic acid ethanolamide,
isostearic acid diethanolamide, monoglyceryl isostearate,
diglyceryl isostearate, stearyl alcohol, stearic acid, stearic acid
ethanolamide, stearic acid diethanolamide, monoglyceryl stearate,
and diglyceryl stearate. Each of these was incorporated into each
test sample. Each of the test samples was prepared in a manner
similar to that of ingredient evaluation test 2. The control
composition employed in ingredient evaluation test 3 was the same
as that employed in Test 2. Each of the test samples comprised 99%
ethanol and the oily ingredient in amounts of 87% and 1%,
respectively.
[0121] In each of the 24 test samples, the amount of pantothenyl
ethylether which was absorbed through skin pores was measured in a
manner similar to that of the above accuracy test. Skin samples
were prepared from the miniature pig's upper back and lower back
(n=5). The results of the evaluation test are shown in FIGS. 7A-7D.
As is apparent from FIGS. 7A-7D, in comparison with the control
composition, in the compositions containing the oily ingredient
having an I.O.B value of 0.06-4.0, the concentration of pantothenyl
ethyl ether delivered to hair follicles increases.
[0122] Measurement Method 1-2
[0123] In order to confirm the validity of the results of the tests
by means of measurement method 1-1, the effect of isostearyl
alcohol on accelerating skin-pore absorption was evaluated by means
of measurement method 1-2 using human hair.
[0124] Thirty male panelists were enrolled in the test. Test
samples (a control composition and a 5% isostearyl
alcohol-containing composition) were prepared as shown in Table 1,
except that the amount of pantothenyl ethylether was changed to 1%.
Each of the test samples (0.5 ml for each) was applied to the scalp
(area: 1 cm.sup.2) of each male panelist, and then the scalp was
left for 16 hours. After the scalp had been washed, 20 hair fibers
were plucked from the scalp of each panelist, to which the test
sample had been applied. The hair roots were cut off the hair
fibers, and the concentration of pantothenyl ethylether delivered
to the hair shafts and outer root sheaths of the hair roots of the
20 hair fibers was measured by means of liquid chromatography under
the same conditions as described above.
[0125] The results of the test are shown in FIG. 8.
[0126] The results obtained through measurement method 1-2 also
show that the concentration of pantothenyl ethylether delivered to
the hair follicles increases in the presence of isostearyl alcohol;
i.e., isostearyl alcohol accelerates the skin-pore absorption of
pantothenyl ethylether.
[0127] 3. Test for Comparison of Isostearyl Alcohol and Transdermal
Absorption-Accelerating Agent
[0128] The test was carried out by means of measurement method 1-1
using a specific amine oxide known as a transdermal
absorption-accelerating agent for comparison.
[0129] In the comparison test, the procedure of the above-described
test by means of measurement method 1-1 was repeated, except that
different test samples were employed and that the concentrations of
.beta.-glycyrrhetinic acid and minoxidil delivered to the hair
follicles were measured by means of liquid chromatography (under
the same conditions as described above). Formulations of the
respective test samples are shown in Table 3. TABLE-US-00003 TABLE
3 Isostearyl Amine Control alcohol oxide composition composition
composition Minoxidil 0.5 0.5 0.5 .beta.-Glycyrrhetinic acid 0.5
0.5 0.5 Ethanol 75 70 70 Ion-exchange water 22.91 22.91 22.91
Isostearyl alcohol -- 5 -- Lauryl dimethylamine oxide -- -- 5
Sodium oleate 0.1 0.1 0.1 Sodium 0.49 0.49 0.49
dodecylbenzenesulfonate Polyoxyethylene 0.5 0.5 0.5 hydrogenated
castor oil (40 E.O.) Total 100 100 100
[0130] The results of the test are shown in FIGS. 9A and 9B. FIG.
9A shows the concentration of .beta.-glycyrrhetinic acid delivered
to the hair follicles, and FIG. 9B shows the concentration of
minoxidil delivered to the hair follicles. The results clearly show
that isostearyl alcohol accelerates the skin-pore absorption of
active ingredients (.beta.-glycyrrhetinic acid and minoxidil) as
compared with amine oxide, and that skin-pore absorption has
different mechanism from transdermal absorption.
[0131] 4. Test for Evaluation of Refreshing Agent
[0132] The test was carried out using test samples containing
various terpene compounds known to be refreshing agents. The
procedure of the above-described test by means of measurement
method 1-2 was repeated, except that the test samples containing
terpene compounds were employed. Formulations of the respective
test samples are shown in Table 4. TABLE-US-00004 TABLE 4
Eucalyptus Control Camphor Menthol oil composition composition
composition composition Pantothenyl 0.5 0.5 0.5 0.5 ethylether
Ethanol 71 70 70 70 Ion-exchange 28.5 28.5 28.5 28.5 water Camphor
-- 1 -- -- Menthol -- -- 1 -- Eucalyptus oil -- -- -- 1 Total 100
100 100 100
[0133] The results of the test are shown in FIG. 10. The results
show that only camphor accelerates the skin-pore absorption of
pantothenyl ethylether.
[0134] The procedure of the above-described test by means of
measurement method 1-1 was repeated, except that the test samples
containing terpene compounds were employed. The results are shown
in FIGS. 11A and 11B. FIG. 11A shows the concentration of
pantothenyl ethylether delivered to the hair follicles, and FIG.
11B shows the concentration of pantothenyl ethylether delivered to
the epidermis and the dermis. As is apparent from FIGS. 11A and
11B, only camphor accelerates the skin-pore absorption of
pantothenyl ethylether.
[0135] Measurement Method 2
[0136] The sebum transferability of aminoxidil-containing
composition was evaluated by use of the kit 10 of the present
invention.
[0137] 1. Minoxidil-containing compositions (minoxidil content: 2%)
were prepared, and each of the compositions (20 .mu.l) was employed
in the test. Formulations of the respective compositions are shown
in Table 5. TABLE-US-00005 TABLE 5 Dipropylene Ethyl Cetostearyl
Isostearyl Ethanol glycol C8 monoglyceride lactate alcohol alcohol
99% Alcohol 88 38 82.7 82.7 82.7 82.7 Polyoxyethylene -- -- 0.1 0.1
0.1 0.1 hydrogenated castor oil (60 E.O.) Isostearic acid -- -- 0.2
0.2 0.2 0.2 Ion-exchange 10 10 10 10 10 10 water Dipropylene -- 50
-- -- -- -- glycol Propylene glycol -- -- -- -- -- -- Isostearyl --
-- -- -- -- 5 alcohol Cetostearyl -- -- -- -- 5 -- alcohol Ethyl
lactate -- -- -- 5 -- -- C8 monoglyceride -- -- 5 -- -- --
Minoxidil 2 2 2 2 2 2 Total 100 100 100 100 100 100
[0138] 2. Artificial sebum was prepared. The formulation of the
artificial sebum is as follows. TABLE-US-00006 Ingredient Amount
(wt. %) Triolein 40.0 Palmitoleic acid 26.0 Wax ester 20.0 Squalane
10.0 Palmitic acid 2.0 Cholesterol 2.0 100.0
[0139] The thus-prepared artificial sebum (11.5 ml) was placed in
the sealable receptacle 11.
[0140] 3. A Franz cell (volume: 11.5 ml) was employed as the
receptacle 11, and a syringe was employed for collecting the
artificial sebum from the cell.
[0141] 4. A single-layer silicone film, one surface of which was
subjected to keratinization, was employed as the thin film 12.
[0142] The test was carried out under the aforementioned
conditions. Specifically, each of the compositions (20 .mu.l) was
applied on the thin film, and the resultant film was allowed to
stand at 37.degree. C. for 24 hours. The degree of transfer of
minoxidil to the artificial sebum was evaluated on the basis of the
concentration of minoxidil contained in the sebum after the elapse
of a specified time period.
[0143] The concentration of minoxidil in the artificial sebum was
measured at predetermined points in time by the following
procedure. The artificial sebum (0.5 ml) was collected by use of a
syringe equipped with a needle, and the collected sebum was
stirred. After completion of stirring, methanol (1.0 ml) was added
to the sebum, and the resultant mixture was allowed to stand, and
then the mixture was subjected to centrifugation at 4,000 rpm for
five minutes. Subsequently, the methanol solution (0.5 ml) which
was separated from the mixture through the centrifugation was
collected, and then the solution was diluted with methanol (1 ml).
The thus-diluted solution was employed as a sample. The sample was
subjected to high-performance liquid chromatography (Nanospace
(product of Shiseido Co., Ltd.), column: Capsule Pack C18, eluent:
(CH.sub.3CN:H.sub.2O=20:80)), to thereby quantify the concentration
of minoxidil. The results are shown in FIG. 12.
[0144] The results show that isostearyl alcohol enhances the sebum
transferability of minoxidil.
[0145] 4. Formulation examples of the composition of the present
invention
[0146] Typical formulation examples of the composition of the
present invention are described below. The below-described
formulations are prepared through customary methods. TABLE-US-00007
Ingredient Amount (wt. %) (Formulation 1) Hair-growing agent
Pantothenyl ethylether 0.5 .beta.-glycyrrhetinic acid 0.5 Swertia
herb extract paste 0.5 Nicotinamide 0.5 Vitamin E acetate 0.5
Ethanol 70.0 Ion-exchange water 21.41 Isostearyl alcohol 5.0 Sodium
lauryldimethylamine oxide oleate 0.49 Polyoxyethylene hydrogenated
castor oil 0.5 (40 E.O.) (Formulation 2) Shampoo Sodium cocoyl
methyl taurate 10.0 Cocamide propyl betaine 5.0 Glycerin 1.0 Citric
acid 1.0 Isostearyl alcohol 5.0 Taurine 1.5 Methyltaurine 0.5
Cationized cellulose 0.5 Perfume suitable amount Ion-exchange water
balance (Formulation 3) Rinse-treatment Stearyltrimethylammonium
chloride 1.0 Isostearyl alcohol 3.0 Palmitic acid 1.0 L-Glutamine
0.1 Ion-exchange water balance (Formulation 4) conditioning shampoo
Sodium N-cocoyl-N-methyl taurate 7.0 Cocamide propyl betaine 7.0
N-[3-(Dimethylamino)propyl]octadecanamide 0.1 Isostearyl alcohol
0.25 Ion-exchange water balance (Formulation 5) Gray-hair
prevention agent 95% Ethanol 55.0 Japanese pepper extract 5.0
Nicotinamide 0.5 Hinokitiol 0.2 1,3-Butylene glycol 1.5
Polyoxyethylene hydrogenated castor oil 1.0 (60 E.O.) Isostearyl
alcohol 5.0 Succinic acid suitable amount Perfume suitable amount
Coloring agent suitable amount Ion-exchange water balance
(Formulation 6) Hair-removing agent 95% Ethanol 55.0 Calcium
thioglycolate 5.0 1,3-Buthylene glycol 1.5 Polyoxyethylene
hydrogenated castor oil 1.0 (60 E.O.) Isostearyl alcohol 5.0
Succinic acid suitable amount Perfume suitable amount Coloring
agent suitable amount
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