Note: Descriptions are shown in the official language in which they were submitted.
Method of manufacturing and the use of Cordyceps cicadae mycelia
active substance for preventing and/or improving acute lung injury
BACKGROUND
Technical Field
[0001] The present invention relates to a Cordyceps cicadae
mycelia
active substance, a method for manufacturing said Cordyceps cicadae
mycelia active substance, and the use thereof. In particular, the present
invention relates to a Cordyceps cicadae mycelia active substance for
preventing and/or improving acute lung injury, a method for manufacturing
said Cordyceps cicadae mycelia active substance, and the use thereof in food
or pharmaceuticals.
Description of Related Art
[0002] Among the ten leading causes of death in 2017 according to
Taiwan's Ministry of Health and Welfare, four of them are closely associated
with lung inflammation, namely, malignant neoplasms (ranked no. 1)
including cancers of trachea, bronchus and lung, pneumonia (ranked no. 3),
and chronic lower respiratory diseases (ranked no. 7).
[0003] Among the ten leading causes of death in the United States,
three of them are associated with lung diseases, namely, lung cancer and
bronchogenic carcinoma, chronic respiratory diseases, influenza, and
pneumonia. Statistical analyses further showed that in the U.S., 79 out of
100,000 were inflicted with acute lung injury (ALI), 59 out of 100,000 were
with acute respiratory distress syndrome (ARDS), and that 43% of all deaths
were caused by ALI/ARDS, that is, 75,000 deaths per year. As air pollution
levels continue to increase and life expectancy continues to rise every year,
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it is estimated that the incidence of ALI/ARDS in the U.S. will at least
double
in 25 years.
[0004] ALT and ARDS (more severe than ALI) are acute inflammatory
lung diseases that are frequently seen clinically, both of which may cause
respiratory failure and may be fatal. They are also associated with many
respiratory diseases.
[0005] ALI-causing risk factors fall into two major groups: direct
and
indirect risk factors. Direct risk factors are those which originate in the
lungs,
such as infective pneumonia caused by bacteria or viruses, aspiration of
gastric acid or foreign objects, or lung contusion. On the other hand,
indirect
risk factors are those which do not originate in the lungs, such as sepsis,
chronic use of alcohol and drugs, or transfusion of artificial blood plasma.
Bacterial infection is one of the above-mentioned major risk factors, an
example being infection by Gram-negative bacteria whose envelope is
mainly composed of endotoxins, also known as lipopolysaccharide (LPS).
[0006] Due to the complex mechanism and high case fatality rate of
ALT, there is still a lack of clinically effective drugs that can be used to
manage ALI's case fatality rate, while common treatments of ALT include
mechanical ventilation, beta-2 adrenergic receptor agonist, anticoagulation,
thrombolysis, surfactant therapy, and surgery. Therefore, research on
effective treatments of ALT remains essential for development.
[0007] Cordyceps cicadae, also known as chong hua, tit chan hua,
hil
chan, and chan yong cao, is a flower bud-shaped stroma formed with strains
at the front end of a cicada larva, which is parasitized by a genus of
Cordyceps, a family of Clavicipitaceae fungal spores, and turned into a
bacteria-carrying carcass. It is written in Compendium of Materia Medica,
Herbology of Classified Syndromes and Zhong hua yao heti that Cordyceps
cicadae is mainly used for treating morbid night crying of babies,
palpitation,
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and malaria, as well as dispelling wind and heat from the body and relieving
convulsion. It has been found through component analysis that natural
Cordyceps cicadae fruit bodies and Ophiocordyceps sinensis are composed
in a similar way.
[0008] Recent pharmaceutical research has found that Cordyceps
cicadae is bioactive with effects of immune response regulation, anti-
oxidation, anti-inflammation, neuroprotection, and anti-cancer. However,
there has been no research done on improving ALI using Cordyceps cicadae
mycelia.
SUMMARY
[0009] Provided herein is a Cordyceps cicadae mycelia active
substance and a method of manufacturing said Cordyceps cicadae mycelia
active substance, which can be used for manufacturing a composition
effective in preventing and/or improving acute lung injury. Compared to
conventional drugs and treatments, the method for manufacturing the
fermentation liquid of the Cordyceps cicadae mycelia active substance
disclosed herein is safer and simpler, and the Cordyceps cicadae mycelia
active substance manufactured is more natural and safer, as well as effective
in improving ALT.
[0010] According to one embodiment of the present invention, a
method is provided for manufacturing a Cordyceps cicadae mycelia active
substance for improving AL!. The method comprises the following steps:
[0011] (a) culturing Cordyceps cicadae mycelia in a plate medium
between 15 and 30 C for 1 to 2 weeks;
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[0012] (b) inoculating the Cordyceps cicadae mycelia of step (a)
to a
flask and culturing the mycelia between 15 and 30 C and at a pH of 2 to 6
for 3 to 14 days; and
[0013] (c) inoculating the Cordyceps cicadae mycelia of step (b)
to a
fermenter tank and culturing the mycelia by stirring between 15 and 30 C
and at a pH of 2 to 6 for 3 to 21 days, so as to obtain a Cordyceps cicadae
mycelium fermentation liquid containing Cordyceps cicadae mycelia active
substance.
[0014] In one embodiment, the method for manufacturing a Cordyceps
cicadae mycelia active substance further includes steps of (d): freeze-drying
the Cordyceps cicadae mycelium fermentation liquid and grinding the
freeze-dried product, so as to obtain a Cordyceps cicadae mycelium powder
containing Cordyceps cicadae mycelia active substance.
[0015] In one embodiment, a gas is further fed into the fermenter
tank
of step (c), and the gas comprises air, oxygen, carbon dioxide, helium or a
combination thereof; the pressure of the fermenter tank is 0.5 to 1.0 kg/cm2,
and a gas flow rate is 0.01 to 1.5 VVM.
[0016] In another embodiment of the present invention, a Cordyceps
cicadae mycelia active substance is provided, which is manufactured using
the said method.
[0017] In yet another embodiment of the present invention, a
composition for preventing and/or improving acute lung injury is provided,
which comprises said Cordyceps cicadae mycelia active substance, and a
pharmaceutically acceptable carrier, excipient, diluent or adjuvant.
[0018] In yet another embodiment of the present invention, A use
of
said Cordycep cicadae mycelia active substance is provided, which is used
for manufacturing a composition for preventing and/or improving acute lung
injury.
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[0019] In one embodiment, improving acute lung injury as mentioned
above includes alleviating pathological symptoms of lung inflammation.
[0020] In one embodiment, alleviating pathological symptoms of
lung
inflammation as mentioned above includes restoring the integrity of
damaged or fused alveoli.
[0021] In one embodiment, improving acute lung injury as mentioned
above includes reducing protein leakage response.
[0022] In one embodiment, improving acute lung injury as mentioned
above includes reducing the level of infiltration in inflamed cells.
[0023] In one embodiment, said inflamed cells includes leukocytes,
phagocytes and/or neutrophils.
[0024] For the purpose of further illustrating the above and other
aspects of the present invention, several exemplary embodiments will be
described with reference to the accompanying drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0025] Fig. 1 shows that pathological conditions of ALI can be
improved by administering with freeze-dried powder of Cordyceps cicadae
mycelia by intraperitoneal injection and then administering endotoxins by
nasal inhalation to induce acute lung inflammation.
[0026] Fig. 2 shows that ALI-caused damage of alveolar-capillary
barrier can be improved by administering with freeze-dried powder of
Cordyceps cicadae mycelia by intraperitoneal injection and then
administering endotoxins by nasal inhalation to induce acute lung
inflammation.
[0027] Fig. 3 shows that ALI-caused infiltration in (A)
leukocytes, (B)
phagocytes and (C) neutrophils can be improved by administering with
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freeze-dried powder of Cordyceps cicadae mycelia by intraperitoneal
injection and then administering endotoxins by nasal inhalation to induce
acute lung inflammation.
[0028] Fig.
4 shows that pathological conditions of ALT can be
improved by administering with freeze-dried powder of Cordyceps cicadae
mycelia orally and then administering endotoxins by nasal inhalation to
induce acute lung inflammation.
[0029] Fig.
5 shows that ALT-caused damage of the alveolar-capillary
barrier can be improved by administering freeze-dried powder of Cordyceps
cicadae mycelia orally and then administering endotoxins by nasal
inhalation to induce acute lung inflammation.
[0030] Fig.
6 shows that ALT-caused infiltration in (A) leukocytes, (B)
phagocytes and (C) neutrophils can be improved by administering freeze-
dried powder of Cordyceps cicadae mycelia by orally and then administering
endotoxins by nasal inhalation to induce acute lung inflammation.
DETAILED DESCRIPTION
Example 1: Culturing Cordyceps cicadae mycelia
[0031]
Cordyceps cicadae mycelia of the present invention are obtained
through the following steps: gathering a natural Taiwanese Cordyceps
cicadae strain, separating its mycelium from the fruit body and storing the
mycelia in a plate medium by subculture. The gene sequence of the strain is
confirmed as Cordyceps cicadae by Taiwan Food Industry Research and
Development Institute. This strain of Cordyceps cicadae strains are now
publicly deposited in the Bioresource Collection and Research Center
(BCRC) of Taiwan Food Industry Research and Development Institute with
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BCRC number: MU30106, but the Cordyceps cicadae mycelia active
substance of the present invention is not limited to the substance prepared
from such strains.
[0032] (1) Plate culture: A Cordyceps cicadae mycelium was
inoculated
onto a plate medium and cultured between 15 to 30 C for 1 to 2 weeks (at
25 C for 7 days in this example). The recipe for the medium may include
Potato Dextrose Agar (PDA), carbon sources and nitrogen sources, and is
not specifically limited.
[0033] (2) Culture in a flask: Cordyceps cicadae mycelia were
scraped
from the plate of step (1) and inoculated to a flask. The culture was grown at
110 to 130 rpm in a shaker incubator between 15 to 30 C and at a pH of 2 to
6 for 3 to 14 days (at 120 rpm, 25 C and pH 5 for 7 days in this example).
The recipe of the shaking culture was shown in Table 1:
[0034] Table 1: Recipe for the culture medium
Amount used in this Preferred range of
Ingredient
example (weight%) amount (weight%)
Mixed carbon and nitrogen 1 0.01 to 5
sources
Carbohydrates 2.5 0.01 to 10
Yeast or malt extract 0.8 0.001 to 2
Animal and plant proteins and 0.5 0.01 to 2
hydrolysates thereof
Inorganic salts 0.05 0.0001 to 0.05
[0035] Among the above ingredients, mixed carbon and nitrogen
sources can be cereals (such as wheat flour) or legumes (such as soya bean
powder, mung bean powder, Glycine max powder or cinnamon powder);
carbohydrates can be glucose, fructose, maltose, sucrose and the like; and
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inorganic salts can be magnesium sulfate, dipotassium phosphate, potassium
dihydrogen phosphate, ferric sulfate and the like. It should be noted that the
recipe as shown in Table 1 is only exemplary, and that the ingredients therein
are not specifically limited and can be adjusted according to actual needs or
used in combination with commercially available culture media.
[0036] (3) Culture in a fermenter tank: The culture in the flask
of step
(2) was further inoculated into a fermenter tank and was stirred between 50
to 150 rpm in temperature between 15 to 30 C, the tank pressure between
0.5 to 1.0 kg/cm2 and the pH between 2 to 6. Meanwhile, gas was fed into
the tank at a gas flow rate of 0.1 to 1.0 VVM. The resulting culture was
incubated for 3 to 21 days to obtain a Cordyceps cicadae mycelium
fermentation liquid (in this example, the culture conditions were 25 C, 0.5
kg/cm2, pH 5, 80 rpm and 1.0 VVM (air) for 14 days). The medium used in
the fermenter tank may contain either the same recipe as the culture in a
flask
of step (2) or other appropriate media (the same recipe as step (2) was used
in this example). This Cordyceps cicadae mycelia fermentation liquid
includes Cordyceps cicadae mycelia active substance of the present
invention. The Cordyceps cicadae mycelium fermentation liquid can be
further freeze-dried to obtain a freeze-dried powder of Cordyceps cicadae
mycelia. In this example, 100 L of Cordyceps cicadae mycelium
fermentation liquid was freeze-dried into 3 kg of freeze-dried powder.
[0037] Cordyceps cicadae mycelia active substance can take various
forms including Cordyceps cicadae mycelium fermentation liquid (strains
and clarified liquid), a freeze-dried powder made from fermentation liquid,
freeze-dried powder dissolved in a solvent, or in other formulations. In a
preferred embodiment, the solvent for dissolving the freeze-dried powder is
water, ethanol or a combination thereof In a preferred embodiment, the ratio
of water to ethanol used as the solvent for dissolving the freeze-dried powder
is 1:1. In the following Example 2, subsequent experiments and analyses
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were carried out with the Cordyceps cicadae mycelia active substance in the
form of freeze-dried powder made from the fermentation liquid.
Example 2: Analyzing the use of Cordyceps cicadae mycelium freeze-
dried powder to improve AL!
[0038] One of the major risk factors leading to ALT is Gram-
negative
bacterial infection, whose envelope is mainly composed of endotoxins, also
known as lipopolysaccharide (LPS). Among various pathogens that cause
ALT, LPS is widely considered as the most effective inducer of acute lung
inflammation and the model of this induction is the closest to the acute
inflammation that occurs clinically. Therefore, suppressing LPS-caused lung
inflammation as well as immune-regulatory functions and mechanisms shall
achieve the effect of improving ALT. There have been experiments in which
animal models of endotoxin-induced ALT are employed. See Yunhe Fu et al.
(2017), Protective effect of TM6 on LPS-induced acute lung injury in mice,
SCIENTIFIC REPORTS, 7:572. A mouse model of acute lung inflammation
was established by LPS following the description in the above publication.
LPS 50 lig/ 20 [iL was administered intranasally to induce acute lung injury.
Analyses of changes in pulmonary pathology, total bronchoalveolar lavage
fluid cell counts, protein concentration and various cytokines were then
carried out, so as to assess the improvements that synthesized peptides, such
as cell-permeable TIR domain-derived decoy peptide, had done to ALT. In
this experiment, a mouse model of acute lung inflammation was also
established using LPS. Analyses of changes in pulmonary pathology, total
bronchoalveolar lavage fluid cell counts, protein concentration and various
cytokines were then carried out to assess the improvements that the
Cordyceps cicadae mycelia active substance had done to ALT.
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[0039] BALB/c mice were used in this animal experiment and were
divided into two major groups according to the route of administration:
intraperitoneal administration and oral administration. The two major groups
were further divided into four groups: a control group to which no substance
was administered (Control group), a group to which only LPS was
administered (LPS group), a group to which both LPS and Cordyceps
cicadae mycelia active substance were administered (CC mycelia group),
and a positive control group to which both LPS and Dexamethasone were
administered (DEX group), making a total of eight groups as shown in Table
2 below. There were six mice in each group with a total of 48 mice.
[0040] Table 2: Group description
Route of Group Substance administered
administration
Intraperitoneal Control N/A
LPS LPS
CC mycelia LPS + CC mycelia 0.25 mg/g
weight of mice (6.25 mg of CC
mycelia for a mouse weighing 25g)
DEX LPS + DEX 5 [ig/g weight of mice
(125 pg of DEX for a mouse
weighing 25g)
Oral Control N/A
LPS LPS
CC mycelia LPS + CC mycelia 0.25 mg/g
weight of mice (6.25 mg of CC
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mycelia for a mouse weighing 25g)
(once per day, three times in total)
DEX LPS + DEX 5 p.g/g weight of mice
(125 1.1g of DEX for a mouse
weighing 25g) (once per day, three
times in total)
[0041] In the intraperitoneal administration group, 20 1_, of LPS
was
administered intranasally, and the concentration of LPS is 50 lig per mouse.
Mice were sacrificed 24 hours after administration, and samples were
collected to be analyzed as described below. In the CC mycelia group,
freeze-dried powder of Cordyceps cicadae mycelia was dissolved in a
water/ethanol solvent at a ratio of 1:1. The resulting mixture was
homogenized using sonication and used as the sample to be administered
when it was confirmed that no precipitation was formed. 50 [EL of Cordyceps
cicadae sample was administered intraperitoneally in mice and endotoxins
(LPS) were administered intranasally 30 minutes after the administration of
the Cordyceps cicadae sample using the same methods of administration and
sacrifice as described above. In the DEX group, DEX was dissolved in saline
and 50 lit of the resulting mixture was administered intraperitoneally in
mice. Endotoxins (LPS) were administered intranasally 30 minutes after the
administration of the DEX sample using the same methods of administration
and sacrifice as described above.
[0042] In the oral administration group, 20 tiL of endotoxins
(LPS)
were administered intranasally, and the concentration of LPS is 50 i.tg per
mouse. Mice were sacrificed 24 hours after administration, and samples were
collected to be analyzed as described below. In the CC mycelia group,
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freeze-dried powder of Cordyceps cicadae mycelia was dissolved in a
water/ethanol solvent at a ratio of 1:1. The resulting mixture was
homogenized using sonication and used as the sample to be administered
when it was confirmed that no precipitation was formed. 200 IlL of
Cordyceps cicadae sample was administered in mice via a feeding tube for
three days, once per day. Endotoxins (LPS) were administered intranasally
three days after using the same methods of administration and sacrifice as
described above. In the DEX group, DEX was dissolved in saline and the
resulting mixture was administered in mice via a feeding tube, once per day.
Endotoxins (LPS) were administered intranasally three days after using the
same methods of administration and sacrifice as described above.
[0043] Histopathological observation of lung tissues: the right
lung of
the mice that was not treated with alveolar lavage was taken out and fixated
immediately in formalin. The fixated product was made into paraffin
sections and dyed with conventional hematoxylin and eosin stain (H&E
stain). Histopathological changes were observed using light microscopy.
[0044] Bronchoalveolar lavage (BAL): lung lavage was performed on
the left lung of the mice with 1 mL of PBS three times using endotracheal
intubation to wash out the contents in the alveolar cavity and to obtain BAL
fluid (BALF). The BALF was centrifuged and the precipitated cells were
measured using differential blood cell count. The protein concentration in
the supernatant was determined using Bradford protein assay.
[0045] Measurement of leukocytes count and type using flow
cytometry: flow cytometry is a highly sensitive quantification method
causing only small personal errors for determining white blood cell
differentials in blood and BALF samples, based on antigens with specificity
that are present on the surface of leukocytes. Targets of measurement
include: (1) cell count in BALF; (2) leukocytes (CD45) count; (3)
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phagocytes (CD45+/CD1 lb) count; (4) neutrophils (CD45+/Ly6G+) count;
and (5) macrophage [(CD45+/CD1 lb+) - (CD45+/Ly6G+)] count.
[0046] Statistical method: data are expressed as Mean + standard
deviation (SD) and analyzed using one-way ANOVA. Group differences are
analyzed using an LSD post-hoc test. Results with a p value less than 0.05
are considered statistically significant.
[0047] As shown in Fig. 1, intraperitoneal injection of freeze-
dried
powder of Cordyceps cicadae mycelia proved to be effective in improving
inflammatory lung diseases. No significant changes were observed in the
Control group with most alveolar structure remaining intact. In the LPS
group, pathological changes in the lung were clearly observed with the
alveolar structure being damaged and fused. On the other hand, the lung
diseases of the mice in the CC mycelia and DEX groups clearly improved.
[0048] After intraperitoneal injection, protein leakage response
caused
by endotoxin-induced lung inflammation was analyzed. It was found that
freeze-dried powder of Cordyceps cicadae mycelia was effective in reducing
protein leakage response, as shown in Fig. 2 and Table 3 below. # p <0.05
compared with Control group; * p <0.05 compared with LPS group.
[0049] Table 3: Protein leakage response after intraperitoneal
injection
Group Protein leakage (mg/mL)
Control 2.87+1.02
LPS 9.93+1.52#
CC mycelia 5.39+1.27*
DEX 3.83+1.72*
(n=6)
# Statistical significance compared to the Control group (p<0.05)
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* Statistical significance compared to the LPS group (p<0.05)
[0050] After intraperitoneal injection, infiltration responses of
(A)
leukocytes (CD45) (B) phagocytes (CD45+/CD1 lb+) and (C) neutrophils
(CD45+/Ly6G+) caused by endotoxin-induced lung inflammation were
analyzed. It was found that freeze-dried powder of Cordyceps cicadae
mycelia was effective in reducing infiltration response of leukocytes,
phagocytes, and neutrophils, as shown in Fig. 3 (A to C) and Tables 4-6
below. # p < 0.05 compared with Control group; * p <0.05 compared with
LPS group.
[0051] Table 4: leukocytes infiltration response after
intraperitoneal
injection
Group leukocytes infiltration (105)
Control 0.14 0.06
LPS 3.29 1.95#
CC mycelia 0.46 0.34*
DEX 0.56 0.29*
(n=6)
# Statistical significance compared to the Control group (p<0.05)
* Statistical significance compared to the LPS group (p<0.05)
[0052] Table 5: phagocytes infiltration response after
intraperitoneal
injection
Group phagocytes infiltration (105)
Control 0.07 0.03
LPS 1.62 0.67#
CC mycelia 0.25 0.14*
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DEX 0.27 0.26*
(n=6)
# Statistical significance compared to the Control group (p<0.05)
* Statistical significance compared to the LPS group (p<0.05)
[0053] Table 6: Neutrophils infiltration response after
intraperitoneal
injection
Group Neutrophils infiltration (105)
Control 0.04 0.02
LPS 1.12 0.80#
CC mycelia 0.15 0.10*
DEX 0.16 0.16*
(n=6)
# Statistical significance compared to the Control group (p<0.05)
* Statistical significance compared to the LPS group (p<0.05)
[0054] As shown in Fig. 4, oral administration of freeze-dried
powder
of Cordyceps cicadae mycelia active substance proved to be effective in
improving inflammatory lung diseases. No significant changes were
observed in the Control group with most alveolar structure remaining intact.
In the LPS group, pathological changes in the lung were clearly observed
with the alveolar structure being damaged and fused. On the other hand, the
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lung diseases of the mice in the CC mycelia and DEX groups clearly
improved.
[0055] After oral administration, protein leakage response caused
by
endotoxin-induced lung inflammation was analyzed. It was found that
freeze-dried powder of Cordyceps cicadae mycelia active substance was
effective in reducing protein leakage response, as shown in Fig. 5 and Table
7 below. # p < 0.05 compared with Control group; * p <0.05 compared with
LPS group.
[0056] Table 7: Protein leakage response after oral administration
Group Protein leakage (mg/mL)
Control 2.60 0.24
LPS 6.65 1.18#
CC mycelia 2.48 0.35*
DEX 3.11 0.64*
(n=6)
# Statistical significance compared to the Control group (p<0.05)
* Statistical significance compared to the LPS group (p<0.05)
[0057] After oral administration, infiltration responses of (A)
leukocytes (CD45) (B) phagocytes (CD45+/CD1 1 b+) and (C) neutrophils
(CD45+/Ly6G+) caused by endotoxin-induced lung inflammation were
analyzed. It was found that freeze-dried powder of Cordyceps cicadae
mycelia active substance was effective in reducing infiltration responses of
leukocytes, phagocytes and neutrophils, as shown in Fig. 6 (A to C) and
Tables 8-10 below. # p < 0.05 compared with Control group; * p <0.05
compared with LPS group.
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[0058] Table 8: leukocytes infiltration response after oral
administration
Group leukocytes infiltration (105)
Control 0.38 0.22
LPS 3.17 0.82#
CC mycelia 0.70 0.46*
DEX 0.88 0.25*
(n=6)
# Statistical significance compared to the Control group (p<0.05)
* Statistical significance compared to the LPS group (p<0.05)
[0059] Table 9: phagocytes infiltration response after oral
administration
Group phagocytes infiltration (105)
Control 0.06 0.04
LPS 0.62 0.35#
CC mycelia 0.11 0.10*
DEX 0.08 0.05*
(n=6)
# Statistical significance compared to the Control group (p<0.05)
* Statistical significance compared to the LPS group (p<0.05)
[0060] Table 10: Neutrophils infiltration response after oral
administration
Group Neutrophils infiltration (105)
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Control 0.02 0.01
LP S 0.29 0.09#
CC mycelia 0.04 0.02*
DEX 0.04 0.02*
(n=6)
# Statistical significance compared to the Control group (p<0.05)
* Statistical significance compared to the LPS group (p<0.05)
[0061] The Cordyceps cicadae mycelia active substance of the
present
invention have been proved through the above experiment to be effective in
improving ALT.
Example 3: Preparing composition
[0062] The present invention provides a composition comprising a
Cordyceps cicadae mycelia active substance to be prepared into a
pharmaceutical composition as well as a health food.
[0063] The composition further includes an additive. In a
preferred
embodiment, the additive can be an excipient, preservative, diluent, filler,
absorption enhancer, sweetener, lubricant, thickener or a combination
thereof The excipient can be selected from sodium citrate, calcium
carbonate, calcium phosphate, sucrose or a combination thereof. The
preservative, such as benzyl alcohol, parabens, silicon or a combination
thereof, can prolong the shelf life of pharmaceutical compositions. The
diluent can be selected from water, ethanol, propylene glycol, glycerol or a
combination thereof The filler can be selected from lactose, high molecular
weight polyethylene glycol or a combination thereof. The absorption
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enhancer can be selected from dimethyl sulfoxide (DMSO), laurocapram,
propylene glycol, glycerol, polyethylene glycol or a combination thereof
The sweetener can be selected from Acesulfame K, aspartame, saccharin,
sucralose, neotame or a combination thereof The lubricant can be selected
from magnesium stearate or gum Arabic. The thickener can be cornstarch.
In addition to the additives listed above, other ones may be selected
according to actual needs provided that the pharmaceutical effects of the
composition are not affected.
[0064] The composition can be developed into various products in
the
pharmaceutical industry. In a preferred embodiment, the composition is a
drug, feed, drink, nutritional supplement, dairy product or health food.
[0065] The composition can take various forms to meet the
subject's
needs. In a preferred embodiment, the composition can be in powder, tablet,
granule, suppository, microcapsule, ampoule/ampule, liquid spray or
suppository form.
[0066] The composition of the present invention can be
administered
to an animal or a human. Provided that the effects of the composition are not
affected, it can be made into any dosage forms and administered via an
appropriate route to the animal or human depending on the dosage form.
[0067] When the Cordyceps cicadae mycelia active substance of the
present invention is of dietary use, the form of the composition 1 as
described
below shall be an illustrative example of the Cordyceps cicadae mycelia
active substance.
[0068] Composition 1: Freeze-dried powder of the Cordyceps cicadae
mycelia active substance (20 wt%) was well-mixed with magnesium stearate
used as a lubricant (8 wt%) and silicon used as a preservative (7 wt%). The
resulting mixture was dissolved in pure water (65 wt%) and stored at 4 C
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for future use. The notation "wt%" refers to the proportion of the weight of
each ingredient relative to the total weight of the composition.
[0069] When the Cordyceps cicadae mycelia active substance of the
present invention is of medical use in a liquid form, the form of the
composition 2 as described below shall be an illustrative example of the
Cordyceps cicadae mycelia active substance.
[0070] Composition 2: Freeze-dried powder of the Cordyceps cicadae
mycelia active substance (20 wt%) was well-mixed with sucralose used as a
sweetener (8 wt%), gum Arabic used as a lubricant (7 wt%) and sucrose used
as an excipient (10 wt%). The resulting mixture was dissolved in pure water
(55 wt%) and stored at 4 C for future use. The notation "wt%" refers to the
proportion of the weight of each ingredient relative to the total weight of
the
composition.
[0071] These examples, though disclosed in the above description,
do
not intend to limit the present invention. A person ordinarily skilled in the
art should be able to make appropriate changes to the features described
therein with reference to the above teachings and still achieve the effects as
claimed by the present application. Therefore, the scope of claims protecting
the present invention shall be determined by the appended claims.
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