Note: Descriptions are shown in the official language in which they were submitted.
Anti-aging Chinese herbal medicine composition, preparation
method and use thereof
Field of the invention
The present invention relates to cosmetics field, particularly to an extract
of
Chinese herbal medicines, its preparation and use, as well as a Chinese herbal
medicine composition containing the extract of Chinese herbal medicines having
anti-aging, especially anti-photo-aging effect, its preparation and its use in
cosmetics.
Background of the invention
Studies have indicated that the skin photo-aging is mainly caused by both the
medium-wavelength ultraviolet (UVB, 290-320 nm) together with the
long-wavelength ultraviolet (UVA, 320-400 nm) in the sunlight. Ultraviolet
rays
significantly hasten the aging of human skin, with one-fold higher risk of
skin aging
and 10-year earlier aging occurrence in high-exposure population as compared
with
low-exposure population. The long-term exposure to the ultraviolet rays of
sunlight
results in the photoaging of the skin and can lead to serious cosmetic issues,
therefore
arising professional attentions in recent years. Anti-photo aging products
have been
released one by one; and more related researches are being conducted. However,
the
main functional supportive additives in current anti-photo-aging cosmetics in
domestic and foreign market are biochemical agent and synthesized compound,
anti-photo-aging additives of Chinese herbal medicine having certain
traditional
Chinese medicine theory guidance remained unseen.
Description of the invention
The technical problem to be solved by the present invention is to provide an
extract of Chinese herbal medicines.
The extract of Chinese herbal medicines according to the present invention is
an
extract by water or solvent of starting materials composed of 10-50 parts by
weight of
Saussurea involucrate, 10-50 parts by weight of Cistanche deserticola and 20-
60
parts by weight of Astragalus membranaceus, wherein the said solvent is
selected
from methanol, ethanol, isopropanol, n-butanol, ethylene glycol, 1,2-
propanediol,
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1,3-butanediol, acetone, ethyl acetate or combinations thereof.
In the above formulations, Saussurea involucrate, Cistanche deserticola and
Astragalus membranaceus all have specific anti-aging effects.
In the present invention, Saussurea involucrata refers to the flower, i.e.,
the
medicinal part, of the snow lotus plant. Snow lotus is a herbaceous plant,
belonging to
subgenus Amphilaena, genus Saussurea, family Asteraceae, and mainly comprises
the
whole plant with flowers of Saussurea involucrata Karet Kir, Saussurea
laniceps
Hand.-Mazz., S.gnaphaloides (Boyle) Sch.-Bip., and S.medusa Maxim. Saussurea
involucrata mainly produced in Sichuan, Yunnan, Tibet, Xinjiang, Gansu,
Qinghai,
etc. Upon blooming from June to July, the whole plants thereof are plucked,
cleaned
from soil, dried, cut into pieces and used in the form of crude materials.
Saussurea involucrata can expel the heat, nourish the blood, keep balance
between Yin and Yang and has the tonifying effect, which is called "the
celestial grass
of thousand years" in Daozang more than one thousand years ago. Yijiuyoushihua
recites "Saussurea involucrata, which are grown in the ancient non-thawing
snow
during one thousand years, look like a lotus flower and could be about 3.3
meters in
height. Being used for brewing liquor, it will boast a feeling of spring
scenery. It
means that Yin reaches extreme so that Yang germinates." In Gangmushiyi, it is
recited that "Saussurea involucrata is produced in the northwest of Yili as
well as
Jinchuan and other cold regions where snow persists from the spring through
summer,
and in snow there are grasses similar to lotus with a single stem, which
stands from
snow in a lovely posture."
Cistanche deserticola refers to the dried fleshy stem, i.e., the medical part
of
Cistanche deserticola or Cistanche tubulosa plant having scale leaf. Cistanche
deserticola is the dried fleshy stem having scale leaf of Cistanche
deserticola Y.C. Ma
or Cistanche tubulosa (Schrenk) Wight. It is plucked mostly in spring when the
young sprout not emerging or just emerging and removed the flower, cut into
pieces
and dried. Cistanche deserticola is mainly distributed in Inner Mongolia,
Gansu,
Xianjiang, Qinghai, etc.
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Desert cistanche is first recorded in "Sheng Nong's herbal classic". It is
recorded that "Desert cistanche has sweet, salty taste, is warm in propertyand
can
tonify kidney, benefit essence and blood and moisten the intestines and free
the stool.
It is mainly used to treat the diseases such as impotence, infertility,
metrorrhagia,
soreness and weakness of waist and knees, cold pains and weak muscle and
joint."
Desert cistanche has always been used as an imperial delicacy worshiped by the
western countries thus it is also called Cistanche deserticola. Cistanche
deserticola
can tonify kidney, benefit the blood and moisten the face, adjust the
endocrine,
promote metabolism and strengthen the body. It also has anti-oxidation effect.
It is
said in "Medical Theory" that Cistanche deserticola can nourish marrow,
moisten the
face, prolong life and has great benefits" and "it can supplement the
deficiency of
essence, dark face and internal lesion caused by overexertion.
Astragalus membranaceus is also named as astragalus and is a general name of
both the plants and the Chinese medicinal materials. Astragalus membranaceus
refers to the dried root, i.e., the medical part of plant Astragalus
membranaceus
(Fisch.) Bge. var. mongholicus (Bge.) Hsiao or Astragalus membranaceus
(Fisch.)
Bge.. Astragalus membranaceus is the dried root of legume plant Astragalus
membranaceus (Fisch.) Bge. var mongholicus (Bge) Hsiao or Astragalus
membranaceus (Fisch.) Bge.. It is plucked in spring and autumn and removed the
fibrous root and root head and dried. Astragalus membranaceus is produced in
North, East north, Inner Mongolia and west north in China and mainly produced
in
Shanxi, Heilongjiang, Liaoning and Hebei. There are also some distributions in
Sichuan, former Soviet Union, North Korea and Mongolia. Astragalus
membranaceus (Fisch.) Bge. is mainly distributed in East North, North, Gansu,
Sichuan and Tibet in China. Astragalus membranaceus (Fisch.) Bge. var
mongholicus (Bge.) Hsiao is mainly produced in Inner Mongolia, Shanxi and
Heilongjiang and now it is planted broadly.
The Chinese medicinal materials Astragalus membranaceus has effects on
reinforcing qi and consolidating the exterior, promoting diuresis and
detumescence,
detoxification and apocenosis and promoting granulation. The medical
applications
of Astragalus membranaceus have a history of more than 2000 years. Current
studies
have indicated that Astragalus membranaceus contains saponins, sucrose,
3
polysaccharides, various amino acids, folic acid and various trace elements
such as
selenium, zinc, copper and the like, which have effects on enhancing immune
functions, protecting livers, promoting diuresis, resisting aging, resisting
stress,
reducing blood pressure and wider antibacterial effect. Astragalus
membranaceus has
the effects on tonifying, retaining youthful looks and moisturizing face, and
thus can
enhance the body resistance, anti-oxidize and postpone aging.
In the current prescription, Astragalus membranaceus acts as the monarch drug
by balancing and nourishing qi and blood and enhancing skin immune function,
Cistanche deserticola and Saussurea involucrate act as the minister drug,
wherein
Cistanche deserticola can strengthen the body, supplement the essence and
nourish
the marrow and Saussurea involucrate can nourish the blood, resist photo-aging
and
oxidation. The root, stem and flower of the medical plant are taken together
to resist
the skin aging by selecting the amounts and controlling the compatibility,
selecting
the rational combination ratio of Astragalus membranaceus, Cistanche
deserticola
and Saussurea involucrate in combination with the studies of ancient
cosmetics. It
has comprehensive effects, wherein the composition is more suitable to be used
in the
cosmetics and has better synergy effects of Chinese medicines, compared with a
single Chinese medicine.
Preferably, the extract of Chinese herbal medicines according to the present
invention is an extract by water or solvent of 30 parts by weight of Saussurea
involucrate, 20 parts by weight of Cistanche deserticola and 50 parts by
weight of
Astragalus membranaceus, wherein the said solvent is selected from methanol,
ethanol, isopropanol, n-butanol, ethylene glycol, 1,2-propanediol, 1,3-
propanediol,
1,3-butanediol, acetone, ethyl acetate or combinations thereof.
Preferably, the extract is carried out by a method selected from decoction,
refluxing, soaking, percolation or a combination thereof.
Preferably, the extract according to the present invention can be in a form
selected from solution, powder, ointment, film or cream.
The present invention also provides a Chinese herbal medicine composition
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having anti photo-aging effect, which comprises the above extract.
The extract of Chinese herbal medicines and the Chinese herbal medicine
composition according to the present invention can be used directly to prevent
the
skin from aging, especially from photo-aging, or can be added to the cosmetics
as
functional additives. The extract of Chinese herbal medicines and the Chinese
herbal
medicine composition according to the present invention have anti-aging
effect,
especially anti-photo-aging effect, and can help the skin against the
oxidation,
facilitate the proliferation of dermal fibroblast, especially help the skin
against the
ultraviolet radiation of UVA and UVB, protect the skin, reduce the damage,
prevent
the skin from photo-agingõ without any irritating effect on the skin. Said
cosmetics
can be cosmetic skin cream, eye cream, facial mask, cleansing milk, lotion or
essence.
The present invention also provides a method for preparing the above extractof
Chinese herbal medicines, comprising:
Mixing 10 to 50 parts by weight of Saussurea involucrate, 10 to 50 parts by
weight of Cistanche deserticola and 20 to 60 parts by weight of Astragalus
rnembranaceus, soaking with water and/or solvent for 0.5-1 1-1 and extracting,
wherein
said solvent is selected from methanol, ethanol, isopropanol, n-butanol,
ethylene
glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone, ethyl
acetate or
combinations thereof;
Filtering, extracting the filtering residue again and combining the filtrates;
Concentrating the filtrate to about 2-fold amount of medicinal materials (V/W)
(fold amount refers to the ratio between the volume of the solvent (ml) and
the total
weight of the medicinal materials (g));
Then carrying out alcohol precipitation;
Removing the solvent from the supernatant from the alcohol precipitation.
The product of alcohol precipitation can be formulated into various dosage
forms,
such as solution, powder, ointment, film or cream. Preferably, the alcohol
used for the
alcohol precipitation is selected from methanol, ethanol, isopropanol, n-
butanol,
ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butylene glycol,
acetone, ethyl
acetate or combinations thereof.
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The extracted solution according to the present invention can be formulated
into
a composition with a concentration of crude drug of 50-1000 mg/ml by adding
water.
Embodiments
The present invention will be further described in detail with reference to
the
following examples.
Example 1
Pure Chinese medicinal materials of Saussurea involucrate 10g, Astragalus
membranaceus 60g and Cistanche deserticola 30g for a total weight of 100g were
mixed thoroughly, to which 10-fold amount (fold amount refers to the ratio
between
the volume of the solvent (m1) and the total weight of the medicinal materials
(g),
hereinafter the same) of 75% (v/v) ethanol was added. The mixture was kept for
cold
soaking for 48 h and then filtered. Ethanol was recovered from filtrate; and
the
resultant extracts were dissolved with water to make a solution with a
concentration
of crude drug of 500 mg/ml, thus obtaining extract A in the form of solution.
Example 2
Pure Chinese medicinal materials of Saussurea involucrate 30g. Astragalus
membranaceus 30g and Cistanche deserticola 40g for a total weight of 100g were
mixed thoroughly, to which 10-fold amount of methanol was added. The mixture
was
subjected to refluxing extraction for 2 h, and filtered. Methanol was
recovered from
filtrate; and the resultant extracts were dissolved with water to make a
solution with a
concentration of crude drug of 50 mg /ml, thus obtaining extract B in the form
of
solution.
Example 3
Pure Chinese medicinal materials of Saussurea involucrate 40g, Astragalus
membranaceus 50g and Cistanche deserticola 10g for a total weight of 100g were
mixed thoroughly, and decocted with 10-fold amount of water for about 2 h,
concentrated and filtered. The supernatant was made into a solution with a
concentration of crude drug of 1000 mg /ml by adding water, thus obtaining
extract C
in the form of solution.
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Example 4
Pure Chinese medicinal materials of Saussurea involucrate 30g. Astragalus
membranaceus 40g and Cistanche deserticola 30g for a total weight of 100g were
mixed thoroughly, and decocted with 10-fold amount of water for about 2 h,
concentrated, precipitated after 2-fold amount of ethanol was added, and then
filtered.
Ethanol was recovered from filtrate; and the resultant extracts were dissolved
with
water and filtered to make a solution with a concentration of crude drug of
500 mg/ml,
thus obtaining extract D in the form of solution.
Experiment 1- DPPH experiment
Free radical is a class of chemicals with active reactivity and extremely high
oxidbillity. The radiation of ultraviolet UVA/UVB can generate numerous free
radicals, which exert damage to the body and accelerate aging. DPPH free
radical
scavenging assay was used herein to determine the in vitro anti-oxidation
activity of
the extracts of the examples.
The extracts of the examples were diluted with distilled water to two
concentrations, 1% and 0.5% (v/v), respectively. Two milliliters of each of
the diluted
extracts of the Examples, 0.5 ml of DPPH solution in ethanol at a
concentration of
500 jimol/L, and 1.5m1 water were then sequentially added into a same test
tube and
shaken thoroughly. After being left stand for 30 min; its optical density (OD)
was
measured at 517 nm by using a corresponding mixture liquid as the blank
control. The
scavenging rates for DPPH of the extracts of the examples were calculated
according
to the following formula:
(
The scavenging rate ( % ) = T ¨TO ) *100 %1 ..
C
wherein:
T --the optical density of DPPH + the extracts of the Examples,
TO -- the optical density of the extracts of the examples + solvents,
C -- the optical density of DPPH + solvents.
The results of evaluation on the anti-oxidation ability of the Chinese herbal
medicine composition according to the present invention were shown in Table I.
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Table 1
Concentration Free radical Concentration Radical
Scavenging Rate Scavenging
Rate
35.04% 0.5% 15.25%
Extract A of Example 1 1%
1% 19.92% 0.5% 8.9%
Extract 13 of Example 2
1% 42.89% 0.5% 19.64%
Extract C of Example 3
1% 54.54% 0.5% 20.31%
Extract D of Example 4
The results of the experiments showed that, the extract A of Example 1, the
extract B of Example 2, the extract C of Example 3 and the extract D of
Example 4
have superior antioxidant ability for free radical scavenging in vitro at the
concentrations of both 1% and 0.5%, wherein all the free radical scavenging
rates at
the concentration of 0.5% are greater than 15% and all the free radical
scavenging
rates at the concentration of 1% are greater than 35%, and the effect from the
extract
D of Example 4 is the best.
Experiment 2- proliferation experiment of fibroblast
The fibroblast in the skin is the main cells for collagen synthesis. The large
amounts of proliferation of fibroblast, or the increased ability of fibroblast
for
synthesizing and secreting collagen can all increase the total amounts of
collagen,
thus postponing the skin aging. Hence, studying the effect of promoting
proliferation
of fibroblast is one of the important methods to evaluate the anti-aging
activity of the
sample. The present experiment uses the method of culturing human fibroblast
in vitro
and preliminarily evaluates the anti-aging effect of the Chinese herbal
medicine
composition according to the present invention based on the above skin aging
mechanism.
The prepared cell suspension (formulated into a concentration of 5*103/m1) was
seeded into a 96-well plate. Except the blank control, 20 I extract of
Chinese herbal
medicines of Examples with corresponding concentrations was added to each well
after the cell attachment. 4 concentration groups were set with 6 wells per
group. The
plate was placed into an incubation chamber (37 C and 5% CO2) to incubate for
24H,
then 20 )1.1 MTT solution was added. The plate was continued to incubate in
the
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incubation chamber (37 C and 5% CO2) for 4H. The enzyme panel was taken out,
the
culture solution in each well was absorbed carefully, and the DMSO was added
and
the wells was mix evenly. The optical density (570 nm) is measured by enzyme-
linked
immunometric meter and the relative proliferation rate is obtained by
comparison
with the control.
The results of proliferation activity of the anti-aging Chinese herbal
medicine
composition according to the present invention to the fibroblast were shown in
Table
2.
Table 2
The concentrations of the Chinese herbal 1% 0.1% 0.01%
0.001%
medicine composition according to the present
invention
The proliferation Extract A of Example 1 20.22% 12.44% 8.98% 5.9%
rate for promoting Extract B of Example 2 17.81% 11.50% 7.43% 4.79%
dermal fibroblast Extract C of Example 3 35.62% 27.68% 23% 10.4%
Extract D of Example 4 25.04% 20.44% 13.22% 6.54%
The results of the experiments indicated that extract A of Example 1, extract
B of
Example 2, extract C of Example 3 and extract D of Example 4 all can promote
the
proliferation of the fibroblast and the effect from the extract C of Example 3
is the
best, wherein the proliferation rate of fibroblast is promoted to be 10.4% at
a
concentration of 0.001%.
Experiment 3- Protective effect on the fibroblast against the damage of UV
radiation
Photoaging of the skin is principally caused by the medium-wavelength
ultraviolet rays (UVB, 290-320 nm) together with the long-wavelength
ultraviolet
rays (UVA, 320-400 nm) in the sunlight, and the ultraviolet rays significantly
hasten
the aging of human skin. UVA/UVB radiation will result in the damage and death
of
fibroblasts and hurt the skin, leading to photoaging. The anti-photoaging
effect of the
extract of Chinese herb medicines of the Examples in protecting the cells
against the
damage from UV radiation was investigated herein by UV radiation assay.
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PBS (4.5 ml) was added to 0.5 ml of each of the extracts of the Examples, to
formulate a dilution with a concentration of 1/10 (v/v), which was subjected
to suction
filtration. Then 0.1 ml filtrate was added into 9.9 ml sterile PBS, finally
obtaining a
dilution in a concentration of 1/1000 (v/v). Skin fibroblasts (Fbs) cultured
with
normal DMEM solution were used as the control group, Fbs irradiated with UVA
(5-20J/cm2) / UVB (20-80J/cm2) after being cultured in normal DMEM solution
were
used as the model group, and Fbs co-cultured using the extracts of the
Examples
(0.15mg/m1) along with DMEM solution and then irradiated with UVA/UVB were
used as the extracts-treated group. After ultraviolet treatment for 24 h, the
optical
density (OD value) of each well was measured using a plate reader with CCK-8.
The
cell proliferation rate was calculated as the percentage of the OD value of
each treated
group relative to the OD value of negative control group; each test was run in
triplicate.
The results of the experiments were shown in Table 3 and Table 4.
Table 3 UVA Radiation
Cell proliferation rate P value
Extract A of Example 1 (86.96+4.68)% <0.01
Extract B of Example 2 (79.80+3.80)% <0.01
Extract C of Example 3 (89.25+4.00)% <0.01
Extract D of Example 4 (95.8+9.9)% <0.01
Control group (100+7.6)%
Model group (73.4+2.3)% <0.01
Table 4 UVB Radiation
Cell proliferation rate P value
Extract A of Example 1 (88.27+3.51)% <0.01
Extract B of Example 2 (82.58+4.20)% <0.01
Extract C of Example 3 (90.86+2.85)% <0.01
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Extract D of Example 4 (94.1 6.8)% <0.01
Control group (100 5.4)%
Model group (76.9 3.5)% <0.01
The results of the experiments showed that, all of the extract A of Example 1,
extract B of Example 2, extract C of Example 3 and extract D of Example 4 can
reduce the damage of UV radiation to fibroblasts and allow the cell
proliferation rate
to be recovered above 90%, wherein the effect from the extract D of Example 4
is the
best.
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