CN111096971A - Rubusoside F1Application in preparing medicine for preventing and treating senile dementia - Google Patents

Rubusoside F1Application in preparing medicine for preventing and treating senile dementia Download PDF

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CN111096971A
CN111096971A CN202010067574.6A CN202010067574A CN111096971A CN 111096971 A CN111096971 A CN 111096971A CN 202010067574 A CN202010067574 A CN 202010067574A CN 111096971 A CN111096971 A CN 111096971A
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rubusoside
senile dementia
medicament
preventing
treating senile
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陈剑鸿
王显风
傅若秋
幸海燕
李斌
蔡永青
明月
倪睿
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Chinese Peoples Liberation Army Army Specialized Medical Center
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia

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Abstract

The invention relates to rubusoside F1The application in preparing medicine for preventing and treating senile dementia belongs to the field of medicine technology. In scopolamine-induced mouse learning and memory disorder model test, rubusoside F1Can obviously improve the learning and memory ability of mice with dysmnesia induced by scopolamine and reduce the acetylcholinesterase activity of mice with dysmnesia induced by scopolamine, and in the rat model experiment of senile dementia induced by A β, rubusoside F1Can obviously improve the learning and memory ability of the rat with the senile dementia induced by A β 25-35, and simultaneously improve the quantity, the arrangement compactness and the nuclear morphology of neurons in the brain tissue of the rat with the senile dementia induced by A β 25-351Used for preparing medicine for preventing and treating senile dementia, and broadens the range of rubusosideF1The application range of the method improves the market value of the method.

Description

Rubusoside F1Application in preparing medicine for preventing and treating senile dementia
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to rubusoside F1Application in preparing medicine for preventing and treating senile dementia.
Background
The senile dementia is a progressive neurodegenerative disease with learning and memory disorders, cognitive dysfunction and language disorders as main clinical manifestations and senile plaques, neurofibrillary tangles and neuronal loss as main pathological features, the main pathological features of the senile dementia are that a large amount of senile plaques, neurofibrillary tangles, β -amyloid protein deposition, neuronal vacuolar degeneration and massive loss occur in the brain, China is gradually introduced into aging society, the proportion of 60-year-old people and above is 13.26 percent by the sixth national population general survey in 2010, the 65-year-old people and above account for 8.87 percent, the senile dementia is the leading cause and the leading cause of mental disability of the elderly, the incidence of 65-year-old people is 1-2 percent and the 85-year-old people is 35 percent, 1 AD patient in every 85 in 2050 is predicted, the senile dementia is a serious social problem, the pathological change process of the senile dementia is quite complex, and although various countries invest a great deal in the research of effective treatment medicines for the senile dementia, a great amount of manpower and material resources are required for searching and developing a great benefit, and side effect for preventing and preventing the senile dementia.
Rubusoside F1(Nigaichigoside F1) Belongs to the triterpenoid saponin compounds. Is widely existed in various plants, such as fruits and whole grass of rubus parvifolius and aleppo avens, and is an important natural product. Modern medical research shows that the rubusoside F1 has pharmacological actions of resisting inflammation, relieving pain, resisting fatigue, resisting oxidation and other physiological activities. However, the anti-senile dementia bioactivity of the rubusoside F1 is not reported.
Disclosure of Invention
In view of the above, one of the objectives of the present invention is to provide rubusoside F1The application in preparing the medicine for preventing and treating the senile dementia; the second purpose is to provide a medicine for preventing and treating senile dementia.
In order to achieve the purpose, the invention provides the following technical scheme:
1. rubusoside F1Application in preparing medicine for preventing and treating senile dementia.
2. A medicine for preventing and treating senile dementia is prepared from kurarinone F1As the only active ingredient, or with rubusoside F1And other components for preventing and treating senile dementia.
Preferably, the other component for preventing and treating senile dementia is one of tacrine, donepezil, galantamine, rivastigmine, huperzine A or huperzine A derivative.
Preferably, the medicament further comprises pharmaceutically acceptable adjuvants.
Preferably, the dosage form of the medicament is one of, but not limited to, capsules, oral liquid, granules, tablets or injections.
Preferably, when the medicament is in a capsule dosage form, the preparation method comprises the following steps: herba Duchesneae Indicae glycoside F1Adding PEG 6000, and allowing the rubusoside F to react at 60-70 deg.C1Uniformly dispersing in PEG 6000, freeze drying, grinding, and sieving with 60-120 mesh sieve to obtain rubusoside F1Solid dispersion of said rubusoside F1Mixing the solid dispersion with microcrystalline cellulose, granulating, drying, and making into capsule.
Preferably, when the medicament is an oral liquid, the medicament is prepared by the following method: herba Duchesneae Indicae glycoside F1Dissolving in water, filtering, adding water to desired volume, stirring, and packaging.
Preferably, when the medicament is in a granule dosage form, the preparation method comprises the following steps: rubusoside F1Adding sucrose powder and dextrin, adding ethanol, granulating, drying, and packaging.
Preferably, the first and second liquid crystal materials are,when the medicament is in a tablet dosage form, the preparation method comprises the following steps: rubusoside F1Adding lactose, adding ethanol, granulating, drying at 50 deg.C or below, adding magnesium stearate, mixing, and tabletting.
Preferably, when the medicament is in an injection dosage form, the preparation method comprises the following steps: herba Duchesneae Indicae glycoside F1Adding sodium chloride into water for injection, adjusting pH to 4.0-5.0, adding water for injection to full volume, stirring, fine filtering, packaging, melt sealing, and sterilizing.
The invention has the beneficial effects that: the invention provides rubusoside F1Application of rubusoside F in preparing medicine for preventing and treating senile dementia in scopolamine-induced mouse learning and memory disorder model test1Can obviously improve the learning and memory ability of mice with dysmnesia induced by scopolamine and reduce the acetylcholinesterase activity of mice with dysmnesia induced by scopolamine, and in the rat model experiment of senile dementia induced by A β, rubusoside F1Can obviously improve the learning and memory ability of the rat with the senile dementia induced by A β 25-35, and simultaneously improve the quantity, the arrangement compactness and the nuclear morphology of neurons in the brain tissue of the rat with the senile dementia induced by A β 25-351Used for preparing medicine for preventing and treating senile dementia, and broadens the scope of rubusoside F1The application range of the method improves the market value of the method.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention. The objectives and other advantages of the invention may be realized and attained by the means of the instrumentalities and combinations particularly pointed out hereinafter.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Example 1
Preparation of rubusoside F1Capsule
The rubusoside F is added according to the mass ratio of 1:61Adding into PEG 6000, mixing, and adding into 60 deg.C water bath to obtain rubusoside F1Uniformly dispersing in PEG 6000, freeze drying at-20 deg.C, grinding into fine powder, and sieving with 80 mesh sieve to obtain rubusoside F1Solid dispersion, namely adding the rubusoside F according to the mass ratio of 7:81Mixing the solid dispersion and microcrystalline cellulose, granulating, drying, and encapsulating to obtain capsule with specification of 0.25 g/capsule.
Rubusoside F1Rubusoside F in capsule1Determination of the content
(1) Chromatographic conditions
Octadecylsilane chemically bonded silica is used as filler, acetonitrile-0.1% phosphoric acid aqueous solution (volume ratio of 20:80) is used as mobile phase, detection wavelength is 205nm, theoretical plate number is determined by picrasin F1The peak calculation should not be below 2000.
(2) Preparation of control solutions
Precisely called picrasma picrorhiza glycoside F1And (3) placing 10.1mg of a reference substance into a 10mL volumetric flask, adding methanol to dissolve the reference substance, adding the reference substance to a scale mark to achieve constant volume, and shaking up to obtain the product.
(3) Preparation of test solution
Precisely weighing the rubusoside F prepared in example 110.5g of capsule content is ground, 25mL of methanol is precisely measured, weighed, ultrasonically treated for 15min, placed to room temperature, weighed, the weight is complemented, shaken evenly and filtered, and the subsequent filtrate is taken, thus obtaining the capsule.
The determination method comprises the following steps: precisely sucking 10uL of each of the reference solution and the sample solution, and measuring with liquid chromatograph to obtain picrasma glycoside F prepared in example 11Rubusoside F in capsule1The content is 16 mg/granule.
Example 2
Rubusoside F prepared in example 11Scopolamine induced mice study by capsuleEffect of the model of learning dysmnesia
Grouping and administration:
collecting 60 clean male BALB/c mice with weight of 20 + -2 g, and randomly classifying into normal group, model group, and rubusoside F1Low dose group (80mg/kg), rubusoside F1Medium dose group (160mg/kg) and rubusoside F1High dose group (320mg/kg), 12/group. Rubusoside F1The administration was performed by gavage, 1 time a day for 7 consecutive days, and the mice in the normal group and the model group were given the same volume of distilled water.
Water maze experiment:
the Morris water maze is a circular water pool with the diameter of 100cm and the height of 50cm, the water pool is divided into four quadrants, a platform with the diameter of 8cm and the height of 35cm is arranged in the water pool, and the position is unchanged in the experimental process. After water is injected, the platform is positioned at a position 2cm under water, and white pigment is added into the water to form a hidden platform. In the experiment, the water temperature is kept at about 20 ℃, so that the indoor environment is dark and quiet, a reference object is established in a visible range of a rat standing on a platform, and the position of the reference object is kept unchanged in the research process. The model group and each rubusoside F were administered by intraperitoneal injection 1h before the experiment1Scopolamine (2mg/kg) caused a learning and memory impairment model in the group, while an equal volume of physiological saline was given to the normal group. Performing a positioning navigation test 30min after administration on the 4 th, 5 th and 6 th days, training four quadrants every time, observing and recording the time for the mouse to find the platform in 120s, namely the escape latency, calculating the average escape latency of each group every day, performing a space search test on the 7 th day, specifically, after removing the platform, searching the position of the platform by the mouse through space memory, and recording the times of the mouse crossing the platform in 120 s.
Determination of cholinesterase activity in brain tissue:
after the completion of the water maze experiment, the mice were dislocated and sacrificed, the left half of the brain was taken, blood on the surface of the brain was washed away with ice-cold physiological saline, weighed, made into 10% brain homogenate with physiological saline at a ratio of 1:9 (weight: volume), centrifuged at 4 ℃ at 3000r/min for 10min, and the supernatant was taken. 100mL of the supernatant of the brain homogenate was added with 1mL of 10mM DTNB [5, 5-dithibis (2-dinitrobenzoic) acid ], mixed well, subjected to water bath at 25 ℃ for 8min, added with 200mL of 8mM acetylcholine, mixed well, and left for 15 min. The absorbance values were measured at 412nm and zeroed with distilled water. To eliminate the effect of the sample, a control tube was required for each assay tube, and the standard tube was 1mmol/L acetylcholinesterase. The activity of acetylcholinesterase is expressed in the amount of hydrolyzed acetylcholine per minute per milligram of tissue.
The experimental results are as follows:
1. rubusoside F1Influence on learning and memory ability of scopolamine-induced dysmnesia mice
Mean escape latency was significantly increased on day 6 in the model group compared to the normal group (P)<0.05), the number of times of crossing the platform has no significant difference; rubusoside F compared to model group1Mean escape latency was significantly reduced (P) on day 6 in the medium and high dose groups<0.05), the number of times of crossing the platform has no significant difference, and the results are shown in table 1.
TABLE 1 rubusoside F1Influence on learning and memory ability of scopolamine-induced dysmnesia mice (n-12)
Figure BDA0002376419000000041
Comparison with normal group # P < 0.05; p <0.05 compared to model group.
2. Rubusoside F1Effect on Acetylcholinesterase Activity in scopolamine-induced dysmnesia mice
Compared with the normal control group, the activity of acetylcholinesterase in the brain tissue of the mouse in the model group is obviously improved (P)<0.01); rubusoside F compared to model group1The acetylcholinesterase activity of the medium and high dose groups is obviously reduced (P)<0.05), the results are shown in table 2.
TABLE 2 rubusoside F1Effect on Acetylcholinesterase Activity in scopolamine-induced dysmnesia mice (n ═ 12)
Figure BDA0002376419000000051
# P <0.01 compared to normal group; p <0.05 compared to model group.
Example 3
Rubusoside F prepared in example 11Effect of capsules on A β -induced Alzheimer's disease rat model
Establishment of a β -induced senile dementia model in rats:
dissolving 1mg of A β 25-35 dry powder in 500 μ L sterile physiological saline, diluting to 2 μ g/μ L solution, and placing in CO2Incubating the culture box at 37 deg.C for 72 hr to make it fibrous and aggregate to form A β 25-35 in aggregate state, storing at 4 deg.C for use, injecting Pentobarbital sodium (40mg/kg) into abdominal cavity, removing hair from the top of rat head, fixing head, removing scalp after conventional sterilization, exposing parietal bone, drilling two small holes on parietal bone for injection, and Hippocampus CA1The specific locations of the zones are: with bregma as the zero point, the distance from the midline to bregma is 3.0mm from the back to the front, the distance from the midline to the midline is + -2.0 mm, and the distance below dura mater is 2.6 mm. Namely 3.0mm behind bregma and 2mm beside the left and right sides of midline as puncture points, drilling skull with dental drill, puncturing dura mater with microsyringe, and vertically inserting needle to 2.6mm below dura mater to obtain Hippocampus CA1And (4) a zone. Bilateral hippocampal CA Using a microinjector1Slowly injecting 5 mu L A β 25-35(2 mu g/mu L) into each area, wherein the injection time of each side hippocampus is 5min, then keeping the needle for 5min to prevent the liquid from overflowing, slowly pulling out the needle after the injection is finished, dropwise adding a proper amount of dental adhesive to seal an injection hole, finally suturing an incision, injecting equal amount of physiological saline into a sham operation group, and continuously injecting penicillin into muscles for three days after the operation to prevent infection.
Grouping and administration:
60 cleaning grade male Wistar rats were randomly divided into six groups: normal group, sham operation group, model group, rubusoside F1Low dose group (40mg/kg), rubusoside F1Medium dose group (80mg/kg) and rubusoside F1High dose group (160mg/kg), 10/group. The administration was started on day 4 after the molding operation, once a day for 35 consecutive days. Rubusoside F1The low, medium and high dosage components are respectively administrated in an intragastric administration mode according to the dosage; normal group, falseThe operative and model groups were given equal amounts of saline.
Water maze test:
the Morris water maze is a circular water pool with the diameter of 100cm and the height of 50cm, the water pool is divided into four quadrants, a platform with the diameter of 8cm and the height of 35cm is arranged in the water pool, and the position is unchanged in the experimental process. After water is injected, the platform is positioned at a position 2cm under water, and white pigment is added into the water to form a hidden platform. In the experiment, the water temperature is kept at about 20 ℃, so that the indoor environment is dark and quiet, a reference object is established in a visible range of a rat standing on a platform, and the position of the reference object is kept unchanged in the research process. Starting from 31 days after administration, performing Morris water maze test 30min after administration every day for 5 days in total, performing positioning navigation test for the first 4 days, and performing space search test for the last day.
Positioning navigation test: the positioning voyage test lasted 4 days. The platform was placed in the center of the second quadrant, the rats entered the water sequentially from the four quadrant faces towards the pool wall, and the time it took for the rats to find the platform within 120s, i.e. the Escape Latency (EL), was recorded. If the rat does not find the platform within 120s, the escape latency is 120s, and the rat should be guided to mount the platform. All rats climbing on the platform should stay on the platform for 15s, so that the rats can know that the underwater platform is an escape point and memorize the spatial position of the platform. The mean escape latency, i.e. the mean escape latency, was calculated for each group of rats in the four quadrants per day. The length of the Average Escape Latency (AEL) reflects the learning and memory ability of the rat, and the shorter the AEL, the faster the learning and cognition of the spatial position of the underwater platform of the rat.
Space search test: and performing a space search test 24h after the positioning navigation test is finished. The specific method is to remove the platform, to make the mouse search the platform by memory under the condition of no platform, and to record the platform crossing times of the rat in 120s, the swimming distance in each quadrant and the total distance. The position of the reference, including the experimenter, around the basin should be fixed throughout the test.
Brain tissue pathology experiments:
after the space search test was completed, the rats were anesthetized with sodium pentobarbital, 5mL of blood was taken from the abdominal aorta of the rats, 4 ℃,centrifuging at 3000r/min for 10min, and collecting serum; taking brain tissue from broken head, immediately fixing with 4% paraformaldehyde, and taking tissue block containing hippocampus by the following steps: a coronal tissue block of 5mm thickness was cut between the superior nucleus of the visual cross and the olfactory bulb by inserting a knife from the superior nucleus of the visual cross. Pathological experiment adopts HE staining, and section of rat brain is observed under 100 times visual field with Hippocampus CA1The pathological morphology of regional brain tissue is mainly observed.
The experimental results are as follows:
1. rubusoside F1Effect on A β 25-35-induced Alzheimer's disease rat escape latency
Mean escape latency was significantly increased from day 2 to day 4 in the model group compared to the normal group (P)<0.05). Rubusoside F compared to model group1Mean escape latencies on days 3 and 4 were significantly reduced (P) for the medium and high dose groups<0.05，P<0.01); rubusoside F1The mean escape latency of the low dose group tended to decrease but there were no significant differences, and the results are shown in table 3.
TABLE 3 rubusoside F1Effect on A β 25-35-induced Alzheimer's disease on escape latency in rats (n ═ 10)
Figure BDA0002376419000000061
Figure BDA0002376419000000071
Comparison with normal group # P < 0.05; p <0.05, P <0.01 compared to model groups.
2. Rubusoside F1Effect on A β 25-35-induced Alzheimer's disease rat spatial search test
Both the number of cross-platform times and the distance ratio of the model group were significantly reduced compared to the normal group (P)<0.05，P<0.01). Rubusoside F compared to model group1The number of cross-platform times to distance ratio was significantly increased (P) on day 5 in the high dose group<0.01，P<0.05); rubusoside F1The number of cross-platform times and distance ratio of the medium dose group tend to increase, but the significance is not poorThe results are shown in Table 4.
TABLE 4 rubusoside F1Effect on A β 25-35-induced Alzheimer's disease rat spatial search test (n ═ 10)
Figure BDA0002376419000000072
Comparison with normal group # P < 0.05; p <0.05, P <0.01 compared to model groups.
3. Rubusoside F1Influence on pathological results of brain tissue of rat with senile dementia
Results show Hippocampus CA1Pyramidal cells of the region, model group hippocampal CA compared to normal group1The cells in the region are arranged sparsely and disorderly, and the cell lines are not clear; the pyramidal cells are obviously reduced; the morphological characteristics of cell apoptosis such as neuron soma shrinkage, nuclear fixation shrinkage and small nuclear plasma ratio appear, and the whole cell dyeing is deepened; the typical pathological features of senile dementia, such as ganglion entanglement, vascular amyloidosis, and granular vacuolization, are found. Rubusoside F compared to model group1The medium and high dose groups were improved in terms of neuron number, degree of packing and neuronal nuclear morphology. Rubusoside F1High dose group CA1The cone cell lines are clear, the cells are arranged more closely, the number is increased obviously, and only a small amount of cells are degenerated and necrotized. The low dose group showed no significant improvement compared to the model group.
Example 3
Rubusoside F1Acute toxicity test of
Taking 40 mice with weight of 20 + -2 g, half each, and administering picrasin F to each mouse at a dose of 16g/kg (body weight)1The administration is carried out 3 times per day by oral gavage for 7 days continuously, and the mice are observed whether acute toxic reaction exists.
The results show that the dosage of the rubusoside F is 100 times of the dosage of the rubusoside F in the oral administration of mice (160mg/kg of the dosage in the mice) to human bodies1In the mean time, no death of the mice was observed, and no LD was detected50. The above results show that rubusoside F1Has good safety, and is equivalent to that when the dosage of the medicine for human body is 100 timesThe mice died.
In the invention, besides the rubusoside F1Adding pharmaceutically acceptable adjuvants to make into capsule, or adding rubusoside F1Adding pharmaceutically acceptable adjuvants, and making into oral liquid, granule, tablet, injection, etc.
Finally, the above embodiments are only intended to illustrate the technical solutions of the present invention and not to limit the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that modifications or equivalent substitutions may be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions, and all of them should be covered by the claims of the present invention.

Claims (10)

1. Rubusoside F1Application in preparing medicine for preventing and treating senile dementia.
2. A medicine for preventing and treating senile dementia is characterized in that the medicine uses rubusoside F1As the only active ingredient, or with rubusoside F1And other components for preventing and treating senile dementia.
3. The medicament for treating senile dementia as claimed in claim 2, wherein the other component for treating senile dementia is one of tacrine, donepezil, galantamine, rivastigmine, huperzine A or huperzine A derivative.
4. The medicament for the prevention and treatment of senile dementia as claimed in claim 2, further comprising pharmaceutically acceptable adjuvants.
5. The medicine for preventing and treating senile dementia as claimed in any one of claims 2-4, wherein the dosage form of the medicine is one of but not limited to capsule, oral liquid, granule, tablet or injection.
6. The medicament for preventing and treating senile dementia as claimed in claim 5, wherein the medicament is prepared by the following method when in the form of capsule: herba Duchesneae Indicae glycoside F1Adding PEG 6000, and allowing the rubusoside F to react at 60-70 deg.C1Uniformly dispersing in PEG 6000, freeze drying, grinding, and sieving with 60-120 mesh sieve to obtain rubusoside F1Solid dispersion of said rubusoside F1Mixing the solid dispersion with microcrystalline cellulose, granulating, drying, and making into capsule.
7. The medicament for preventing and treating senile dementia as claimed in claim 5, wherein the said medicament is prepared by the following method when it is in the form of oral liquid: herba Duchesneae Indicae glycoside F1Dissolving in water, filtering, adding water to desired volume, stirring, and packaging.
8. The medicament for preventing and treating senile dementia as claimed in claim 5, wherein the medicament is prepared by the following method when it is in the form of granules: rubusoside F1Adding sucrose powder and dextrin, adding ethanol, granulating, drying, and packaging.
9. The medicament for preventing and treating senile dementia as claimed in claim 5, wherein the said medicament is prepared by the following method when in tablet form: rubusoside F1Adding lactose, adding ethanol, granulating, drying at 50 deg.C or below, adding magnesium stearate, mixing, and tabletting.
10. The medicament for preventing and treating senile dementia as claimed in claim 5, wherein the said medicament is prepared by the following method when it is in the form of injection: herba Duchesneae Indicae glycoside F1Adding sodium chloride into water for injection, adjusting pH to 4.0-5.0, adding water for injection to full volume, stirring, fine filtering, packaging, melt sealing, and sterilizing.
CN202010067574.6A 2020-01-20 2020-01-20 Rubusoside F1Application in preparing medicine for preventing and treating senile dementia Pending CN111096971A (en)

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