A composition for treating senile dementia and a method for preparing the same

Abstract

The present application provides a kind of composition for treating senile dementia and its preparation method.The composition is composed of Cistanche and other multi-flavor Chinese medicinal materials, and is used for treating mild and moderate senile dementia, and is suitable for popularization and application.

Description

Technical Field

[0001] This invention relates to a clinically experienced medication for treating Alzheimer's disease, exploring the pathogenesis and treatment of Alzheimer's disease from the perspective of "kidney essence deficiency," and achieving significant therapeutic effects in clinical application. It belongs to the field of traditional Chinese medicine application. Background Technology

[0002] Alzheimer's disease falls under the categories of "dementia," "forgetfulness," and "dullness" in Traditional Chinese Medicine (TCM). Clinically, it is characterized by cognitive decline and forgetfulness. Previous studies have attributed the disease to factors such as old age, physical weakness, overwork, loneliness, and emotional imbalance. From the perspective of organ dysfunction, the kidneys, spleen, and heart are often considered to be involved, with pathogenesis involving essence, qi, blood, phlegm, stagnation, and blood stasis. It is believed that old age and physical weakness lead to qi deficiency and blood deficiency, resulting in insufficient qi to transport blood, causing blood stasis, which in turn obstructs qi flow. Qi stagnation leads to phlegm obstruction, and the accumulation of phlegm and blood stasis obstructs the brain's pathways, ultimately resulting in dementia. However, Alzheimer's disease primarily occurs in the elderly, and its progression is closely related to age. The key pathogenesis is the depletion of essence in old age, particularly kidney essence deficiency, gradual depletion of marrow, malnourishment of the brain, and impaired mental function. Therefore, exploring the pathogenesis and pathogenesis of Alzheimer's disease from the perspective of kidney essence deficiency is of significant importance and value.

[0003] Traditional Chinese medicine's theory of collaterals posits that collaterals are divided into qi collaterals (which circulate qi) and blood collaterals (which circulate blood), each fulfilling the normal physiological functions of "qi warming and blood nourishing." The *Qianjin Yaofang* states, "The head is the origin of the body, the abode of the spirit, the source of qi, blood, and essence; all 365 collaterals ascend to the head," highlighting the close connection between collaterals and the brain. The qi collaterals of the brain play a role in warming and nourishing, transmitting information, and regulating and controlling, encompassing the higher central nervous system's functions of thinking, movement, and language. The blood collaterals of the brain, on the other hand, function in supplying blood and qi, exchanging body fluids and blood, and facilitating nutrient metabolism, encompassing various levels of small and medium-sized blood vessels and microvessels, especially the microcirculation, that branch off from the vertebral artery and internal carotid artery throughout the brain. Normal cognitive and memory functions of the brain depend on unobstructed blood vessels. If one is old and has depleted essence, the functions of the five internal organs gradually decline, the vaporization and transformation of qi become abnormal, and the body fluids cannot be transported and distributed, resulting in phlegm and turbidity. Alternatively, if the kidney essence is insufficient to transform qi and is unable to transport blood, blood vessels will be blocked. Phlegm and blood stasis will accumulate and cause obstruction of blood vessels, which will damage the brain and spirit. The brain marrow will be deprived of nourishment and wither, which will aggravate mental disorders. The *Ling Shu* (Spiritual Pivot), in its chapter "On the Nine Needles," states, "The reason humans are born is through blood vessels." This indicates that the ends of the blood vessels play a crucial role in supplying blood and qi, exchanging body fluids and blood, and facilitating nutrient metabolism. When blood vessels are unobstructed, the qi and blood vessels function normally, and the brain's cognitive and memory functions are normal. The *Ling Shu*, in its chapter "On the Absence of Grains in Normal People," states, "When blood vessels are smooth and unobstructed, the spirit resides." The kidneys store primordial essence, which transforms into primordial qi. If one is old and weak, primordial qi is insufficient, and the qi in the blood vessels is depleted, resulting in weak blood circulation and obstructed blood flow. As Wang Qingren said, "When primordial qi is deficient, it cannot reach the blood vessels. When the blood vessels lack qi, they will stagnate and become stagnant." The *Reading Notes on Medicine*, in its chapter "On the Inheritance of Life and Transformation," also clearly points out, "When qi is deficient, it is insufficient to propel blood, and blood will inevitably stagnate." It is pointed out that when Qi is deficient, the body's ability to propel Qi or warm it is weakened, leading to dysfunction in the blood and Qi supply, fluid exchange, and nutrient metabolism at the extremities. Phlegm is the stagnation of fluids, and blood stasis is the stagnation of blood. Since fluids and blood share the same origin, phlegm and blood stasis are related. Therefore, they can further combine to obstruct the meridians, causing structural and functional abnormalities in the Qi channels and resulting in malnourishment of the brain meridians. Obstruction in the meridians damages the meridians and accelerates the formation of phlegm and blood stasis. Phlegm and blood stasis, as secondary pathogenic factors, obstruct the meridians, causing dysfunction in the blood and Qi supply and fluid exchange at the extremities, which will eventually cause or aggravate damage to the Qi channels. It can be seen that deficiency of kidney essence and Qi and malnourishment of the brain meridians are the key pathogenesis of dementia, which then develops into phlegm and blood stasis obstructing the meridians, further aggravating the condition.

[0004] Based on the above etiology and pathogenesis, the treatment of this disease should focus on tonifying the kidneys and replenishing essence. In addition, promoting blood circulation and improving intelligence are of great significance for accurately grasping the pathogenesis.

[0005] Another patent of the inventor, CN111939237A, discloses a composition for assisting in improving memory, consisting of Acanthopanax senticosus, ginseng, Alpinia oxyphylla, and Schisandra chinensis. However, the treatment principle of this invention is different from that of the present invention. This patent uses ginseng as the principal ingredient, focusing on replenishing qi and promoting body fluid production, with an emphasis on improving learning and memory impairments. Summary of the Invention

[0006] Based on the above understanding, kidney essence deficiency and insufficient marrow are the main pathogenesis of senile dementia. The treatment principle of "tonifying the kidney and replenishing essence, unblocking the meridians and improving intelligence" is established. By replenishing kidney essence to strengthen the foundation, the kidney essence is sufficient, the marrow is nourished, the brain meridians are unblocked, and the mind can be used effectively. This can significantly improve symptoms such as memory loss, slow thinking, soreness of the waist and knees, dizziness, tinnitus, fatigue and soreness, and frequent urination at night caused by kidney essence deficiency.

[0007] Based on the TCM pathogenesis characteristics of Alzheimer's disease, namely "deficiency of kidney essence and insufficiency of marrow sea", the treatment principle is "tonifying the kidney and replenishing essence, unblocking the meridians and improving intelligence". Cistanche deserticola is the principal drug, Acanthopanax senticosus is the assistant drug, Alpinia oxyphylla is the adjuvant drug, and Angelica sinensis is the guiding drug. The formulation of the drug composition of this invention is established.

[0008] The principal herb is Cistanche deserticola, which is sweet, sour, and salty, warm in nature, and enters the kidney meridian. It was first mentioned in the Shennong's Classic of Materia Medica, which states that it "treats the five types of fatigue and seven types of injury, replenishes the middle jiao...nourishes the five internal organs, regulates yin, and benefits essence and qi." It has the functions of tonifying the kidney, replenishing essence, and benefiting marrow. Cistanche deserticola replenishes kidney essence, is warm but not hot, tonifying but not drastic, and warming but not drying. It is chosen as the principal herb because of its nature of nourishing and replenishing kidney essence and its gentle and warming properties, so that the marrow sea can be nourished and the spirit can be utilized.

[0009] The assistant herb, Acanthopanax senticosus, is pungent, slightly bitter, and warm in nature, entering the spleen, kidney, and heart meridians. It tonifies the kidneys, strengthens the spleen, replenishes qi, and nourishes the spirit. As described in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica), it is listed as a superior herb, stating that "long-term use can lighten the body, prolong life, and is harmless." Simultaneously, it replenishes qi and strengthens the spleen. Combined with the principal herb, Cistanche deserticola, it enhances the effect of tonifying the acquired constitution and protecting the innate essence. It also invigorates blood circulation and unblocks the meridians, addressing pathological products such as phlegm and blood stasis that obstruct the meridians, thus promoting the flow of qi, blood, and essence to the brain, resulting in abundant energy and restored memory.

[0010] Assistant herb: Alpinia oxyphylla, pungent, warm, enters the spleen and kidney meridians. It tonifies the kidney and consolidates essence, promotes yang and dispels yin to replenish insufficient kidney essence, warms the kidney and consolidates essence, and stores and returns to its source. It is effective for those with intellectual decline, forgetfulness, lethargy and reluctance to sleep caused by insufficient kidney essence. When used with the principal herb, it enhances the effects of replenishing essence and marrow, nourishing essence and consolidating the kidney.

[0011] Guiding herb: Angelica sinensis, which is warm in nature and sweet and pungent in taste, enters the heart, liver, and spleen meridians. It nourishes and invigorates blood, moistens dryness and lubricates the intestines. It is good at entering the blood and serves as a guiding herb to lead other herbs into the collaterals. Angelica sinensis can both nourish and invigorate blood, and can both unblock the meridians and activate the collaterals. It nourishes while moving and moves while nourishing, making it an essential herb for the blood. When combined with Acanthopanax senticosus, it enhances the effect of activating blood and unblocking collaterals, leading other herbs into the blood and activating blood to unblock the collaterals. It has a better effect on improving the symptoms of forgetfulness and insomnia in elderly and weak patients.

[0012] In summary, the drug composition of the present application is aimed at the basic pathogenesis of kidney essence deficiency in senile dementia, and the treatment focuses on filling kidney essence, dredging collaterals and benefiting intelligence. The prescription composition selects Cistanche and Acanthopanax for filling kidney essence and tonifying the body, and simultaneously uses Fructus Alpiniae Oxophyllae to tonify kidney essence, selects Angelica and Acanthopanax to activate blood and dredge collaterals, and the prescription is simple and efficient. The drug composition not only helps to improve the dementia performance caused by kidney essence deficiency, such as reduced intelligence and frequent forgetfulness, but also significantly improves the clinical symptoms caused by kidney essence deficiency in patients with senile dementia, such as dizziness, tinnitus, tooth shaking and falling, lumbago, leg weakness, walking difficulty, laziness and lying, and improves the quality of life.

[0013] The composition is composed of the following weight parts of traditional Chinese medicinal materials:

[0014] Cistanche 200-600 parts, Acanthopanax 400-1200 parts, Fructus Alpiniae Oxophyllae 200-600 parts, and Angelica 200-600 parts.

[0015] The composition is preferably prepared from the following weight parts of raw medicinal materials:

[0016] Cistanche 300-500 parts, Acanthopanax 600-1000 parts, Fructus Alpiniae Oxophyllae 300-500 parts, and Angelica 300-500 parts.

[0017] The composition is preferably prepared from the following weight parts of raw medicinal materials:

[0018] Cistanche 400 parts, Acanthopanax 800 parts, Fructus Alpiniae Oxophyllae 400 parts, and Angelica 400 parts.

[0019] The composition is preferably prepared from the following weight parts of raw medicinal materials:

[0020] Cistanche 380 parts, Acanthopanax 700 parts, Fructus Alpiniae Oxophyllae 400 parts, and Angelica 500 parts.

[0021] The composition is preferably prepared from the following weight parts of raw medicinal materials:

[0022] Cistanche 600 parts, Acanthopanax 400 parts, Fructus Alpiniae Oxophyllae 500 parts, and Angelica 500 parts.

[0023] The active component of the drug composition of the present application is prepared by the following steps:

[0024] (1) Raw material treatment: crushing Fructus Alpiniae Oxophyllae;

[0025] (2) Weighing: weighing Cistanche, Acanthopanax, Fructus Alpiniae Oxophyllae and Angelica according to the prescription ratio for standby;

[0026] (3) Take Cistanche, Acanthopanax, 50% -80% ethanol reflux extraction 2-3 times, 1-3 hours each time, add 8-10 times each time, filter the extract, filter the filtrate, and reduce the pressure to concentrate to a clear paste with a relative density of 1.05~1.10 (60 ℃), and reserve;

[0027] (4) Take Fructus Schisandrae Chinensis, Angelica, add 8-12 times the amount of water, heat distillation for 6-10 hours, collect the volatile oil; water decoction is ready for use; the residue is added with 8-12 times the amount of water, decocted twice, each time for 1.5 hours, the two decoctions are combined with the water decoction after distillation, and reduced pressure concentration to a clear paste with a relative density of 1.05~1.10, combined with the clear paste obtained in step (3), and continued to reduce pressure concentration to a thick extract with a relative density of 1.20-1.30;

[0028] (5) Take the thick extract obtained in step (4) and dry it, crush it, and the dry extract powder and the volatile oil together constitute the active ingredient of the pharmaceutical composition of the invention.

[0029] The Cistanche in the composition is Cistanche tubulosa.

[0030] The pharmaceutical composition can be prepared into capsule, tablet, pill, powder or paste.

[0031] The preparation method of the tablet is as follows:

[0032] (1) Raw material processing: break Fructus Schisandrae Chinensis;

[0033] (2) Weighing: take Cistanche, Acanthopanax, Fructus Schisandrae Chinensis, Angelica according to the prescription proportion;

[0034] (3) Take Cistanche, Acanthopanax, 50% -80% ethanol reflux extraction 2-3 times, 1-3 hours each time, add 8-10 times each time, filter the extract, filter the filtrate, and reduce the pressure to concentrate to a clear paste with a relative density of 1.05~1.10 (60 ℃), and reserve;

[0035] (4) Take Fructus Schisandrae Chinensis, Angelica, add 8-12 times the amount of water, heat distillation for 6-10 hours, collect the volatile oil; water decoction is ready for use; the residue is added with 8-12 times the amount of water, decocted twice, each time for 1.5 hours, the two decoctions are combined with the water decoction after distillation, and reduced pressure concentration to a clear paste with a relative density of 1.05~1.10, combined with the clear paste obtained in step (3), and continued to reduce pressure concentration to a thick extract with a relative density of 1.20-1.30; dry and crush to obtain dry extract powder;

[0036] (5) After the volatile oil is encapsulated, it is dried and crushed into fine powder;

[0037] (6) Mix the dry extract powder of step (4) with the fine powder of step (5), add excipients, and prepare tablets.

[0038] The present application also provides the use of the composition in improving memory disorders.

[0039] The present application also provides the use of the composition in improving cholinergic system function, reducing oxidative stress damage, reducing neural cell apoptosis or inhibiting inflammatory response.

[0040] The Latin name of the raw material drug as the active component in the composition of the present application and its processing method are from "Chinese Medicine Dictionary" (July 1977, first edition, Shanghai Scientific and Technical Press) and "Chinese Pharmacopoeia" (2005 edition, Chemical Industry Press).

[0041] The pharmaceutical composition of the present application can be prepared into any conventional dosage form acceptable in pharmacy by conventional preparation process, for example, the preparation process described in "Pharmacy of Chinese Medicine" (Shanghai Scientific Press, December 1997, first edition) to form any conventional dosage form acceptable in pharmacy, such as capsules, tablets, pills, powders, soft capsules or ointments, etc.

[0042] In the application of the present application, the pharmaceutical composition is one of capsule, tablet, pill, powder, soft capsule or ointment preparation. In order to realize the above dosage forms, pharmaceutically acceptable excipients, such as fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives, bases, etc. are added during the preparation of these dosage forms. The fillers include starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, etc.; the disintegrants include starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, cross-linked sodium carboxymethyl cellulose, etc.; the lubricants include magnesium stearate, sodium dodecyl sulfate, talc, silicon dioxide, etc.; the suspending agents include polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methyl cellulose, etc.; the binders include starch paste, polyvinylpyrrolidone, hydroxypropyl methyl cellulose, etc.; the sweeteners include sodium saccharin, aspartame, sucrose, cyclamate, glycyrrhetinic acid, etc.; the flavoring agents include sweeteners and various flavors; the preservatives include nipagin, benzoic acid, sodium benzoate, sorbic acid and its salts, benzalkonium bromide, chlorhexidine acetate, eucalyptus oil, etc.; the bases include PEG6000, PEG4000, insect wax, etc. In order to realize the above dosage forms, other pharmaceutically acceptable excipients (the excipients described in each dosage form in "Pharmacy of Chinese Medicine", Shanghai Scientific Press, December 1997, first edition) are added during the preparation of these dosage forms.

[0043] Aiming at the basic pathogenesis change of kidney essence deficiency of senile dementia, the treatment focuses on filling kidney essence, dredging collaterals and benefiting intelligence. The prescription composition selects Cistanche deserticola and Acanthopanax senticosus to supplement kidney and essence, and simultaneously uses Yizhi Ren to fill kidney essence, and uses Angelica sinensis and Acanthopanax senticosus to activate blood and dredge collaterals. The prescription has simple and efficient composition, which not only helps to improve the dementia performance caused by kidney essence deficiency, such as intelligence decline, forgetfulness and the like, but also significantly improves the clinical performance caused by kidney essence deficiency, such as dizziness, tinnitus, tooth shaking, hair loss, waist soreness, leg weakness, walking difficulty, laziness and sleepiness, and improves the life quality.

[0044] To confirm the effect of the composition of the present application on senile dementia, the following pharmacological tests were carried out using the preparation prepared according to the method of Example 1 and Example 2 (hereinafter referred to as RJYZ1 and RJYZ2).

[0045] Test 1:

[0046] The effect of the pharmaceutical composition of the present application on Aβ 42 oligomer-induced Alzheimer's disease rats

[0047] ABSTRACT

[0048]

Objective

Methods

Results

Conclusion

[0049] Abbreviation list

[0050] Alzheimer's disease (AD)

[0051] Aβ β-amyloid protein β-amyloid protein

[0052] ACHE Acetylcholinesterase

[0053] MDA malondialdehyde

[0054] SOD superoxide dismutase

[0055] p-Tau phosphorylation-Tau

[0056] Bcl-2 B cell lymphoma / lewkmia-2

[0057] Bax Bcl-2 Assaciated X protein

[0058] Caspase-3 cysteinyl aspartate-specific proteinase

[0059] IL-6 interleukin-6

[0060] IL-1β interleukin-1β

[0061] IL-18 interleukin-18

[0062] TNF-α tumor necrosis factor-α

[0063] APP β-amyloid precursor protein

[0064] Ach acetylcholine

[0065] Experimental purposes

[0066] AD model was induced by injecting Aβ oligomer into the CA1 region of the hippocampus of rats 42 The learning and memory improving effects of the pharmaceutical composition of the present application on AD rats were evaluated.

[0067] 1 Experimental materials

[0068] 1.1 Test sample

[0069] 1.1.1 Name: Drug composition 1 of the invention, abbreviation: RJYZ1, drug composition 2 of the invention, abbreviation: RJYZ2, control drug, capsule content prepared according to the patent embodiment 1 of CN111939237A.

[0070] 1.1.2 Property: Brown powder.

[0071] 1.1.3 Clinical indication: Tonifying kidney and filling essence, dredging collaterals and benefiting intelligence. Senile dementia.

[0072] 1.1.4 Clinical dosage: Oral, the clinical dosage for human is 24.0g crude drug per day.

[0073] 1.1.5 Content and specification: 5.0g crude drug / g dry paste powder.

[0074] 1.1.6 Source and batch number: Source: Hebei Yiling Hospital, batch number: 20150406.

[0075] 1.1.7 Storage condition: Sealed.

[0076] 1.2 Positive drug, tool drug and main reagent

[0077] Aβ 42 Oligomer: Synthesized by the Institute of Biological Science and Technology, School of Science, Beijing Jiaotong University.

[0078] Huperzine A tablet: Henan Tailong Pharmaceutical Co., Ltd., batch number: 140902.

[0079] Anti-brain decline capsule: Shijiazhuang Four Pharmaceutical Co., Ltd., batch number: KN15031001.

[0080] 10% formaldehyde buffer solution: Beijing Yili Fine Chemicals Co., Ltd., batch number: 20150504.

[0081] SOD detection kit: Nanjing Jiancheng Biological Engineering Institute, batch number: 20150821.

[0082] MDA detection kit: Nanjing Jiancheng Biological Engineering Institute, batch number: 20150826.

[0083] ACHE detection kit: Nanjing Jiancheng Biological Engineering Institute, batch number: 20150820.

[0084] BCA protein quantification kit: Beijing Solabio Technology Co., Ltd., batch number: 20151015.

[0085] Aβ primary antibody: abcam, batch number: ab39377.

[0086] Tau primary antibody: abcam, batch number: ab32057.

[0087] p-Tau primary antibody: abcam, batch number: ab131354.

[0088] Bcl-2 primary antibody: abcam, batch number: ab7973.

[0089] Bax primary antibody: abcam, batch number: ab7977.

[0090] Caspase-3 primary antibody: abcam, batch number: ab44976.

[0091] IL-6 primary antibody: abcam, batch number: ab83339.

[0092] IL-1β primary antibody: abcam, batch number: ab9722.

[0093] IL-18 primary antibody: abcam, batch number: ab191860.

[0094] TNF-α primary antibody: abcam, batch number: ab6671.

[0095] 1.3 Experimental system

[0096] 1.3.1 Animal strain: SD rats.

[0097] 1.3.2 Animal level: SPF level.

[0098] 1.3.3 Animal gender and quantity: half male and half female, a total of 120.

[0099] 1.3.4 Animal age: 7 weeks old.

[0100] 1.3.5 Animal weight: 180-210g.

[0101] 1.3.6 Animal source: purchased from Beijing Vantoll Life Science and Technology Co., Ltd., qualified certificate number: 11400700105927, license number: SCXK (Jing) 2012-0001, received date July 22, 2015.

[0102] 1.3.7 Raising conditions: The animals were raised in the New Drug Evaluation Center of Hebei Province of Integrative Medicine Research Institute. The rats were raised in cages, with 12 hours of light per day, temperature 20-26℃, relative humidity 40-70%.

[0103] 1.3.8 Quarantine process: New animal quarantine period for 5 days, during the quarantine period, the animals had normal drinking and eating, good health, no disease and death signs.

[0104] 1.3.9 Feed: experimental grade granular rat feed, provided by Jiangsu Province Coordinated Medicine and Biological Engineering Co., Ltd., production license: Su Shifa Shen (2014) 01008.

[0105] 1.3.10 Drinking water: fill with ordinary water for animals to drink freely, flush the drinking water bottle and change the water once a day.

[0106] 1.3.11 Bedding: ordinary grade bedding for experimental animals, provided by Hebei Province Experimental Animal Center, production license: SCXK (Hebei) 2013-2-001.

[0107] 1.3.12 Identification: 5% picric acid marking.

[0108] 2 Experimental method

[0109] 2.1 Experimental design basis

[0110] 2.1.1 Standard adopted: Chinese Ministry of Health Drug Administration Bureau issued "Guidelines for the Study of New Drugs of Traditional Chinese Medicine (Pharmacy, Pharmacology, Toxicology)", "Compilation of Preclinical Research Guidelines for New Drugs (Western Medicine) (Pharmacy, Pharmacology, Toxicology)", "Methodology of Traditional Chinese Medicine Pharmacological Research", "Methodology of Pharmacological Experiments" published by People's Medical Publishing House and related literature.

[0111] 2.1.2 Information provided by the entrusting unit: RJYZ1, RJYZ2, the proposed clinical human dose of the control drug group is 24.0g of crude drug per day, and the human body is calculated according to 60kg body weight, so the proposed clinical dose is 0.4g of crude drug per kg, and the content of the test product dry powder is 5.0g of crude drug per g of dry powder, batch number 20150406.

[0112] 2.2 Dose and grouping

[0113] Rats were randomly divided into 8 groups according to body weight, namely blank group, sham operation group, model group, huperzine A group, anti-brain failure group, RJYZ1 group, RJYZ2 group, and control drug group, male and female, 3.2g of crude drug / kg per day -1 , equivalent to 8 times the clinical human dose, the maximum daily dose of huperzine A (9 tablets) is 450μg per person, and the maximum daily dose of anti-brain failure capsules (18 capsules) is 32.02g of crude drug per person. The dose of rats is 8 times the maximum daily dose, which is 60μg / kg per day -1 and 4.3g of crude drug / kg per day -1 . See Table 1.

[0114]

[0115] 2.3 Method of administration

[0116] Gavage administration, consistent with the clinical recommended oral route.

[0117] 2.4 Preparation and storage of test articles

[0118] The test drugs were prepared into experimental concentrations with distilled water (see Table 1), and stored at 2-8°C after preparation for standby use. The positive drugs were prepared on the spot.

[0119] 2.5 Administration of test articles

[0120] The animals were administered by gavage at 10 ml / kg, and the blank, sham operation and model groups were given distilled water, once a day for 5 weeks.

[0121] 2.6 Steps, methods and detection indexes

[0122] 2.6.1 Step: The newly received animals were labeled, and quarantined for 5 days. After quarantine, 12 animals in the blank group and 14 animals in the sham operation group were randomly selected according to body weight, with half male and half female. The remaining animals were injected with 1 μg / μl Aβ 42 oligomers 5 μl, and the sham operation group was injected with the same amount of normal saline. The blank group was not treated. One week after the operation, the remaining animals were divided into groups according to body weight, i.e., the model group, huperzine A group, anti-brain failure group, and RJYZ low, medium and high dose groups, each with 14 animals. After the grouping was completed, gavage administration was started. The blank, sham operation and model groups were given distilled water, and the body weight was recorded once a week for 5 weeks. After 3 weeks of administration, the learning and memory ability test, i.e., the water maze behavioral test, was performed. After the behavioral test, 3 animals were randomly taken from each group, perfusion fixed and taken out of the brain for standby use. After the administration was completed, the remaining animals were anesthetized with 10% chloral hydrate, and blood was taken from the abdominal aorta. After centrifugation at 3000 rpm for 10 min, the serum was taken for blood biochemical detection. The bilateral hippocampus was taken on ice, stored in liquid nitrogen, and 3 animals were prepared for immunoblotting detection, and the rest were homogenized for tissue biochemical detection.

[0123] 2.6.2 Methods and detection indexes

[0124] 2.6.2.1 Operation method: The rats were anesthetized by intraperitoneal injection of 10% chloral hydrate, and the head was fixed on a stereotaxic instrument. The scalp was shaved and disinfected, a longitudinal incision was made on the back of the head, the skull was exposed, and the bilateral hippocampal CA1 region was selected as the injection site according to the Rat Brain Stereotaxic Atlas written by Professor Paxinos of the University of New South Wales in Australia and the Rat Brain Stereotaxic Atlas edited by Bao Xinmin et al. The coordinates were as follows: 3.5 mm behind the bregma, 2.1 mm on both sides of the midline, and 3.6 mm under the dura. The skull was drilled with a dental drill ring at the coordinate point, the needle was slowly inserted vertically to the target point, and then 5 μl (1 μg / μl) Aβ 42Oligomer constant speed slow injection, injection time 5 min, needle 3 min, slow needle 3 min, to ensure that the solution is fully dispersed, the sham group was given the same amount of saline, the rest of the operation was the same as before, a little penicillin powder was given at the incision, and the incision was sutured.

[0125] 2.6.2.2 Water maze behavioral method: Water maze experiment is mainly used to test the spatial location learning and memory ability of experimental animals, which is composed of a circular pool, a platform and a video tracking system. The circular pool is divided into 4 quadrants. This time, two tests are involved, including positioning navigation test (4d) and spatial exploration test (1d).

[0126] Positioning navigation test: the platform is fixed at the center of the first quadrant 2 cm underwater, the water temperature is controlled at 22-26℃, at the opposite side of the platform away from the platform, two water entry points equidistant from the platform are randomly selected, the animal is gently placed in the water facing the pool wall, the maximum swimming time is set to 90s, if the rat finds the platform within 90s, it is allowed to stay on the platform for 10s, if not, it is artificially placed on the platform for 10s, twice a day for 3 consecutive days, and the average time (escape latency) and total distance of each group of rats to find the platform from the two water entry points on the 4th day are recorded.

[0127] Spatial exploration test: after the positioning navigation test, the platform is removed, and then the rat is placed in the water at any same water entry point, and the rat's stay time in the first quadrant and the number of times crossing the original platform position within 60s are recorded.

[0128] 2.6.2.3 Biochemistry, immunoblotting

[0129] Rats were anesthetized, blood was collected from the abdominal aorta and centrifuged to obtain serum, blood biochemistry detection, bilateral hippocampus was taken on ice, liquid nitrogen preservation, 3 animals were prepared for immunoblotting detection, and the rest were homogenized for tissue biochemistry detection.

[0130] Hippocampal tissue biochemistry detection: ACHE, SOD, MDA.

[0131] Hippocampal tissue immunoblotting detection: Aβ, Tau, p-Tau, Bcl-2, Bax, Caspase-3, IL-6, IL-1β, IL-18, TNF-α.

[0132] Blood biochemistry detection: SOD, MDA.

[0133] 2.7 Related staff notification

[0134] When purchasing animals, notify the animal room, and when the animal appears abnormal, notify the pathology room for treatment.

[0135] 2.8 Main instrument system

[0136]

[0137] 2.9 Statistical Methods

[0138] Experimental data were analyzed using SPSS 11.5 statistical software. Statistical results are expressed as mean ± standard deviation. The method involves first performing a normality test. For data that conforms to a normal distribution, one-way ANOVA is used to compare the means. If the variances are homogeneous, the least significant difference (LSD) method is used for pairwise comparisons. If the variances are not homogeneous, Dunnett's T3 test is used for pairwise comparisons. If the data does not conform to a normal distribution, nonparametric tests are used for statistical analysis.

[0139] 3 Results

[0140] 3.1 Effects of RJYZ on learning and memory abilities in AD rats.

[0141] As shown in Table 2, compared with the sham surgery group, the latency and total distance of the water maze navigation experiment in the model group were significantly increased (P<0.01), while those in the RJYZ1 group, RJYZ2 group, and control drug group were significantly decreased compared with the model group (P<0.01). Compared with the sham surgery group, the number of platform crossings and the time spent in the platform quadrant in the space exploration experiment in the model group were significantly decreased (P<0.01), while those in the RJYZ1 group, RJYZ2 group, and control drug group were significantly increased compared with the model group (P<0.01 or P<0.05). This indicates that injection of Aβ into the hippocampus... 42 Oligomers cause learning and memory impairments in animals, while RJYZ can improve learning and memory abilities.

[0142] 3.2 Effects of RJYZ on the cholinergic nervous system function of AD rats.

[0143]

[0144] As shown in Table 3, compared with the sham-operated group, the hippocampal ACHE activity in the model group rats was significantly increased (P<0.01), while the hippocampal ACHE activity in the RJYZ1 group, RJYZ2 group, and control group was significantly decreased compared with the model group (P<0.01 or P<0.05). This indicates that RJYZ can indirectly increase ACh content by reducing hippocampal ACHE activity, thereby improving the cholinergic nervous system function and enhancing learning and memory abilities in AD rats.

[0145] 3.3 Effects of RJYZ on oxidative stress capacity in AD rats.

[0146] As shown in Table 3, compared with the sham operation group, the MDA content in the hippocampus of the model group was significantly increased (P<0.05), and the MDA content in the hippocampus of the RJYZ1 group, the RJYZ2 group and the control group showed a decreasing trend, although there was no significant difference (P>0.05) compared with the model group. There was no significant difference in the SOD activity in the hippocampus and the MDA content in the serum among the groups (P>0.05). It can be seen that RJYZ can reduce the oxidative stress damage by reducing the MDA content in the hippocampus, and improve the learning and memory ability.

[0147]

[0148] 3.4 Effect of RJYZ on the expression of Aβ in the hippocampus of AD rats.

[0149] As shown in Table 4, compared with the sham operation group, the expression of Aβ in the model group was significantly increased (P<0.01), and the expression of Aβ in the RJYZ1 group, the RJYZ2 group and the control group was significantly decreased (P<0.01) compared with the model group. It can be seen that RJYZ can improve the learning and memory ability by reducing the deposition of Aβ in the hippocampus of AD rats.

[0150] 3.5 Effect of RJYZ on the expression of p-Tau in the hippocampus of AD rats.

[0151] As shown in Table 4, compared with the sham operation group, the expression of p-Tau in the model group was significantly increased (P<0.01), and the expression of p-Tau in the RJYZ1 group, the RJYZ2 group and the control group was significantly decreased (P<0.01 or P<0.05) compared with the model group. It can be seen that RJYZ can improve the learning and memory ability by reducing the overexpression of p-Tau in the hippocampus of AD rats, reducing the neuronal degeneration and necrosis caused by neurofibrillary tangles.

[0152] 3.6 Effect of RJYZ on apoptosis in the hippocampus of AD rats.

[0153] As shown in Table 4, compared with the sham operation group, the expression of Caspase-3 in the model group was significantly increased (P<0.01), and the expression of Caspase-3 in the RJYZ1 group, the RJYZ2 group and the control group was significantly decreased (P<0.01) compared with the model group. The Bcl-2 / Bax ratio in the RJYZ1 group and the RJYZ2 group was significantly increased (P<0.01 or P<0.05) compared with the model group. It can be seen that RJYZ can protect the normal function of cells by reducing the expression of the apoptosis protein Caspase-3 in the hippocampus of AD rats and increasing the Bcl-2 / Bax ratio, and improve the learning and memory ability.

[0154] 3.7 Effect of RJYZ on inflammation in the hippocampus of AD rats.

[0155] As shown in Table 4, compared with the sham-operated group, the expression of IL-6, IL-1β, IL-18, and TNF-α in the model group was significantly increased (P<0.01 or P<0.05). The expression of IL-6 in the RJYZ1 group, RJYZ2 group, and control group was significantly decreased compared with the model group (P<0.01). The expression of IL-1β, IL-18, and TNF-α in the RJYZ1 group and RJYZ2 group was significantly decreased compared with the model group (P<0.05). This indicates that RJYZ can reduce the expression of inflammatory factors, alleviate the inflammatory response, protect normal cell function, and improve learning and memory abilities.

[0156] 4. Conclusion

[0157] RJYZ against Aβ 42 Oligomers significantly improve learning and memory abilities in AD rats. This effect may be achieved by reducing ACHE activity, improving cholinergic system function, alleviating oxidative stress damage caused by increased MDA content in the hippocampus, reducing Aβ deposition in the hippocampus, reducing neurofibrillary tangles caused by p-Tau overexpression in the hippocampus, reducing the expression of apoptotic protease Caspase-3, increasing the anti-apoptotic Bcl-2 / Bax ratio, reducing neuronal apoptosis, and inhibiting the expression of inflammatory factors IL-6, IL-1β, IL-18, and TNF-α, thereby reducing the inflammatory response.

[0158]

[0159] Experiment 2:

[0160] Effects of the pharmaceutical composition of the present invention on an ethanol-induced mouse model of memory recall deficit

[0161] summary

[0162] [Objective] To observe the effect of the pharmaceutical composition of the present invention on improving memory ability in mice with memory recall deficit induced by ethanol. [Methods] Seventy-two ICR mice, half male and half female, were randomly divided into six groups according to body weight: blank group, model group, positive drug group, RJYZ1 group, RJYZ2 group, and control drug group. The positive drug was huperzine A, with a maximum daily clinical dose of 450 μg / d in humans. The dosage for mice was 10 times the maximum daily clinical dose, i.e., 75 μg / kg × day. -1The animals are continuously administered by gavage for 10 days, the blank and model groups are given the corresponding volume of solvent, 1h after the last administration, the step platform experiment training is started, training for 5min, 24h later, the memory performance is tested, the model group and each administration group is given 40% ethanol 10ml / kg by gavage 30min before testing, the blank group mice are given the corresponding volume of distilled water by gavage, the latency period of the first jump down platform and the number of errors within 5min are observed and recorded as memory indicators.

Results

Conclusions

[0163] Experimental purpose

[0164] The ethanol-induced memory reproduction deficiency model of mice is used to observe the improvement effect of the pharmaceutical composition of the application on the learning and memory ability of the memory reproduction deficiency mice.

[0165] 1 Experimental materials

[0166] 1.1 Test sample (the same as the test sample in test 1)

[0167] 1.2 Positive drugs, tool drugs and main reagents

[0168] Huperzine A tablets: Henan Tailong Pharmaceutical Co., Ltd., batch number: 140902.

[0169] 95% ethanol: Tianjin Fugui Chemical Industry Co., Ltd., batch number: 131028.

[0170] Picric acid: Taishan Yiao Reagent Plastic Co., Ltd., batch number: 20131001.

[0171] 1.3 Experimental system

[0172] 1.3.1 Animal strain: ICR mice.

[0173] 1.3.2 Animal level: SPF level.

[0174] 1.3.3 Animal gender and quantity: half male and half female, a total of 72.

[0175] 1.3.4 Animal age: 4 weeks old.

[0176] 1.3.5 Animal weight: 16~20g.

[0177] 1.3.6 Animal source: purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., qualified certificate No. 1140700091998, license No. SCXK (Jing) 2012-0001, received on April 22, 2015.

[0178] 1.3.7 Raising conditions: The animals were raised in the New Drug Evaluation Center of Hebei Province of Integrative Medicine Research Institute. The light was 12 hours / day, the temperature was 20~26℃, and the relative humidity was 40~70%. The mice were caged, 6 / cage.

[0179] 1.3.8 Quarantine process: The quarantine period of new animals was 3 days. During the quarantine period, the animals had normal drinking and eating, good health, and no signs of disease and death.

[0180] 1.3.9 Feed: Experimental level granular mouse feed, provided by Jiangsu Province of Collaborative Medicine Biological Engineering Co., Ltd., production license: Su Shifa (2014) 01008.

[0181] 1.3.10 Drinking water: Filling ordinary water for animals to drink freely, washing the drinking water bottle and changing the water once a day.

[0182] 1.3.11 Bedding: Experimental animal ordinary level bedding, provided by Hebei Province Experimental Animal Center, production license: SCXK (Ji) 2013-2-001.

[0183] 1.3.12 Identification: 5% picric acid marking.

[0184] 2 Experimental method

[0185] 2.1 Experimental design basis

[0186] 2.1.1 Standard: The Research Guide for New Drugs of Traditional Chinese Medicine (Pharmacy, Pharmacology, Toxicology) issued by the Drug Administration Bureau of the Ministry of Public Health of the People's Republic of China, Preclinical Research Guidelines for New Drugs (Western Medicine) Compilation (Pharmacy, Pharmacology, Toxicology), Pharmacological Research Methodology of Traditional Chinese Medicine, Pharmacological Experimental Methodology published by People's Medical Publishing House and related literature materials.

[0187] 2.1.2 Information provided by the entrusting unit: The proposed clinical dose of RJYZ for human use is 24.0g of crude drug per day. According to the calculation of 60kg body weight, the proposed clinical dose is 0.4g of crude drug per kg, and the content of the test product dry powder is 5.0g of crude drug per g of dry powder, batch No. 20150406.

[0188] 2.2 Dose and grouping

[0189] The mice were randomly divided into 6 groups according to body weight, namely, blank group, model group, positive drug (huperzine A) group, RJYZ1 group, RJYZ2 group, and control drug group, 12 mice in each group, and the RJYZ1 group, RJYZ2 group, and control drug group were set to 4.0 crude drug / kg x day -1 , which is equivalent to 10 times of the clinical human dose, and the maximum daily dose of huperzine A in clinic (9 tablets) is 450 μg / person / day, and the mouse dose is 10 times of the maximum daily dose in clinic, that is, 75 μg / kg x day -1 . See Table 5.

[0190]

[0191] 2.3 Administration method

[0192] Gavage, consistent with the recommended oral route in clinic.

[0193] 2.4 Preparation and storage of test products

[0194] Each test drug was prepared into the experimental concentration (see Table 5) with distilled water, and after preparation, it was stored at 2-8°C for standby, and the positive drug was prepared immediately before use.

[0195] 2.5 Administration of test products

[0196] The animals were administered by gavage at 10 ml / kg, and the blank group and model group were given distilled water, once a day for 10 consecutive days.

[0197] 2.6 Steps and observation indexes

[0198] The mouse jumping platform device is six rectangular reflection boxes, with a size of 32 cm * 22.5 cm * 33 cm, which is divided into 6 rooms by a black plastic plate, with a stainless steel grid at the bottom, and the grid spacing is 0.5 cm. A rubber platform with a height of 4.5 cm in diameter is placed at the left rear corner of each room. The mouse is placed on the platform facing the corner, and it is adapted to the environment for 3 min, and then a 36V current is passed through. If the animal jumps off the platform, it will be shocked. Its normal response should be to jump back to the platform to avoid the harmful stimulus. Most animals may jump onto the grid again or multiple times to escape the shock. When jumping down, the mouse's both feet touch the copper grid at the same time, which is considered as a mistake. The training lasts for 5 min, and the test is conducted after 24 h. This is a memory retention test. The latency period and the number of errors in the first jump off the platform within 5 min are recorded.

[0199] Newly received animal label, quarantine 3d, after quarantine, the animals were randomly divided into 6 groups according to body weight, as described above. Intragastric administration according to body weight, once a day, for 10 days, the blank group and the model group were given distilled water, 1h after the last administration, start the platform training, training for 5min, 24h later test memory performance, the model group and each administration group were given 40% ethanol 10ml / kg by gavage 30min before testing, the blank group mice were given distilled water of the corresponding volume by gavage, the latency and error times of the first jump off the platform within 5min were observed and recorded, the latency of the mice that did not jump off within 5min was calculated as 300s.

[0200] 2.7 Related staff notification

[0201] When purchasing animals, inform the animal room, and inform the pathology room for processing when the animals have abnormal conditions.

[0202] 2.8 Instrument system

[0203] DT2000 electronic balance, Changshu Shuangjie Testing Instrument Factory.

[0204] SL-2001N electronic balance, Shanghai Minqiao Precision Scientific Instrument Co., Ltd.

[0205] Stopwatch, Shenzhen Huibuo Trading Co., Ltd.

[0206] DT200 mouse platform instrument, Chengdu Tailian Technology Co., Ltd.

[0207] 2.9 Statistical method

[0208] Experimental data were analyzed and processed by SPSS11.5 statistical software, and the statistical results were represented by mean ± standard deviation (). Firstly, normality test was performed, for data conforming to normal distribution, mean comparison was performed by one-way ANOVA, if the variance was equal, pairwise comparison was performed by least significant difference method (LSD), if the variance was not equal, Dunnett's T3 test was used for pairwise comparison; if not conforming to normal distribution, non-parametric test was used for statistical analysis.

[0209] 3 Results

[0210] As shown in Table 6, compared with the blank group, the latency of the model group mice was significantly shortened (P<0.05), and the error times were significantly increased (P<0.01), and the two indicators showed that the ethanol modeling was successful; compared with the model group, the latency and error times of the control drug group mice had no significant difference (P>0.05), the latency of the RJYZ2 group mice was significantly prolonged (P<0.05), and the error times were significantly reduced (P<0.05), the RJYZ1 group also showed that the latency was significantly prolonged (P<0.05), and the error times were significantly reduced (P<0.01).

[0211]

[0212] 4 Conclusion

[0213] The experimental results show that the pharmaceutical composition has certain improvement effect on the memory reproduction deficiency of mice caused by ethanol.

[0214] 5 Discussion

[0215] Learning and memory disorder is an important manifestation of senile dementia, and is one of the important brain functions indispensable for human and animal survival. Learning is the process of obtaining new behavior and experience by the nervous system in response to changes in the external environment, and memory is the retention and reproduction of experience after learning. Generally, learning and memory are divided into three stages, namely, acquisition, consolidation and reproduction. Ethanol is a central nervous system inhibitor and is a commonly used tool drug for memory reproduction deficiency model. It can inhibit the conditioned reflex process of animals, hinder the synthesis of brain protein and RNA, and change the cholinergic and dopamine systems, thereby destroying the learning and memory function.

[0216] The platform test is a commonly used behavioral method for testing learning and memory, but its accuracy is easily affected by many factors, such as experimental environment, experimental animals, non-specific interference, reward and punishment effect, etc.

[0217] In this experiment, the experience accumulated in the previous stage of the laboratory was used for reference. The food intake of the animals was controlled in the experiment to prevent the body weight from being too large, reduce the influence of difficulty in getting on the platform due to excessive body weight, and maintain a quiet experimental environment with suitable and consistent indoor temperature, humidity and light. The results of the experiment showed that the latency and error times of the traditional Chinese medicine compound RJYZ had significant differences compared with the model group, and had certain dose dependence, indicating that the traditional Chinese medicine compound RJYZ had certain improvement effect on the memory reproduction deficiency model of mice caused by ethanol.

[0218] Test 3:

[0219] Effect of the pharmaceutical composition of the application on the memory consolidation disorder model of mice caused by sodium nitrite

[0220] ABSTRACT

[0221] [Objective] To observe the improvement effect of the pharmaceutical composition of the application on the learning and memory ability of memory consolidation disorder mice by using the memory consolidation disorder model of mice caused by sodium nitrite. [Methods] 96 ICR mice, half male and half female, were randomly divided into 6 groups according to body weight: blank group, model group, positive drug group, RJYZ1 group, RJYZ2 group and control drug group. The dosages of RJYZ1 group, RJYZ2 group and control drug group were 4.0g of crude drug / kg / day -1, which is equivalent to 10 times of the clinical human dose, and the positive drug is Huprine, the maximum daily dose of which in human is 450 μg / d, and the dose of the mouse is 10 times of the maximum daily dose, i.e. 75 μg / kg / day -1 . The animals were continuously administered intragastrically for 10 days according to the body weight, the blank and model groups were administered with the corresponding volume of solvent, 1 h after the last administration, the model group and each administration group were immediately administered with subcutaneous injection of sodium nitrite 120 mg / kg, the volume was 10 ml / kg, the blank group of mice was administered with the corresponding volume of normal saline, and the memory performance was tested after 24 h, and the latency period and the number of errors within 5 min of the first time of jumping down the platform were observed and recorded as the memory index.

Results

Conclusions

[0222] Experimental purposes

[0223] The memory consolidation disorder model of mice was used to observe the improvement effect of the pharmaceutical composition of the application on the learning and memory ability of the memory consolidation disorder mice.

[0224] 1 Experimental materials

[0225] 1.1 Test sample (the same as the test sample in test 1)

[0226] 1.2 Positive drug, tool drug and main reagent

[0227] Huperzine A tablet: Henan Tailong Pharmaceutical Co., Ltd., batch number: 140902.

[0228] Sodium nitrite: Tianjin Bodu Chemical Co., Ltd., batch number: 20131113.

[0229] Picric acid: Taishan Yiao Reagent Plastic Co., Ltd., batch number: 20131001.

[0230] 1.3 Experimental system

[0231] 1.3.1 Animal strain: ICR mice.

[0232] 1.3.2 Animal level: SPF level.

[0233] 1.3.3 Animal gender and number: half male and half female, 96 in total.

[0234] 1.3.4 Animal age: 4 weeks old.

[0235] 1.3.5 Animal weight: 16-20 g.

[0236] 1.3.6 Animal source: purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., certificate number: 11400700095243, license number: SCXK (Jing) 2012-0001, received on May 13, 2015.

[0237] 1.3.7 Raising conditions: the animals were raised in the New Drug Evaluation Center of Hebei Province of Integrative Medicine Research Institute. The mice were kept in cages with 12 hours of light per day, a temperature of 20-26°C, and a relative humidity of 40-70%.

[0238] 1.3.8 Quarantine process: new animal quarantine period of 5 days, during the quarantine period, the animals had normal drinking and eating, good health, no disease and death signs.

[0239] 1.3.9 Feed: experimental grade granular mouse feed, provided by Jiangsu Synergy Pharmaceutical Biological Engineering Co., Ltd., production license: Su Shifa (2014) 01008.

[0240] 1.3.10 Drinking water: ordinary water was filled for animals to drink freely, and the drinking water bottles were washed and replaced once a day.

[0241] 1.3.11 Bedding: ordinary grade bedding for experimental animals, provided by Hebei Province Experimental Animal Center, production license: SCXK (Ji) 2013-2-001.

[0242] 1.3.12 Identification: 5% picric acid marking.

[0243] 2 Experimental methods

[0244] 2.1 Experimental design basis

[0245] 2.1.1 Standard adopted: Chinese Ministry of Health Drug Administration Bureau promulgated "Guidelines for the Study of New Drugs of Traditional Chinese Medicine (Pharmacy, Pharmacology, Toxicology)", "Compilation of Preclinical Research Guidelines for New Drugs (Western Medicine) (Pharmacy, Pharmacology, Toxicology)", "Methodology of Traditional Chinese Medicine Pharmacological Research", "Methodology of Pharmacological Experiments" published by People's Medical Publishing House and relevant literature materials.

[0246] 2.1.2 The information provided by the entrusting unit: the proposed clinical dose of RJYZ is 24.0 g crude drug per day, and the proposed clinical dose for a 60 kg human is 0.4 g crude drug per kg. The content of the dry powder of the test product is 5.0 g crude drug per g dry powder, and the batch number is 20150406.

[0247] 2.2 Dose and grouping

[0248] The mice were randomly divided into 6 groups according to body weight, i.e. a blank group, a model group, a positive drug (huperzine A) group, a low, a medium and a high dose group of RJYZ, with 16 mice in each group. The dose of RJYZ in groups 1 and 2 and the control drug group was determined to be 4.0 g crude drug per kg per day, which is 10 times the clinical human dose. -1 The maximum daily dose of huperzine A in clinical use is 450 μg per person per day (9 tablets), and the dose for mice is 10 times the maximum daily dose in clinical use, i.e. 75 μg per kg per day. -1 See Table 7.

[0249]

[0250] 2.3 Administration method

[0251] Gavage, consistent with the recommended oral route in clinical use.

[0252] 2.4 Preparation and storage of test product

[0253] The test drugs were prepared into the experimental concentration (see Table 7) with distilled water, and stored at 2-8°C after preparation for future use. The positive drug was prepared immediately before use.

[0254] 2.5 Administration of test product

[0255] The animals were administered by gavage at a dose of 10 ml / kg, and the blank and model groups were administered with distilled water. The administration was performed once a day for 10 consecutive days.

[0256] 2.6 Procedure and observation indexes

[0257] The mouse jumping platform device is six rectangular reflective boxes, with a size of 32 cm * 22.5 cm * 33 cm, separated into six rooms by black plastic plates. The bottom is a stainless steel grid with a grid spacing of 0.5 cm. A rubber platform with a height of and a diameter of 4.5 cm is placed in the left rear corner of each room. The mice are placed gently on the platform with their faces towards the corner, and they are allowed to adapt to the environment for 3 minutes. Then a current of 36 V is applied, and if the animals jump off the platform, they will be shocked. Their normal response should be to jump back onto the platform to avoid the harmful stimulus. Most animals may jump onto the grid again or multiple times to escape the shock. When jumping off, the mice are considered to have made a mistake if both their feet touch the copper grid at the same time. The training lasts for 5 minutes, and the test is performed 24 hours later. This is a memory retention test, and the latency period and the number of errors in the first jump off the platform within 5 minutes are recorded.

[0258] Newly received animal label, quarantine 5d, after quarantine, the animals were randomly divided into 6 groups according to body weight, as described above. Intragastric administration according to body weight, once a day, for 10 days, the blank group and the model group were given distilled water, 1 h after the last administration, the platform training was started, training for 5 min, after training, the model group and each administration group were immediately injected subcutaneously with sodium nitrite 120 mg / kg, the volume was 10 ml / kg, the blank group mice were given the corresponding volume of normal saline, 24 h later, the memory score was tested, and the latency period and the number of errors of the first jump down platform within 5 min were observed and recorded, the jump platform latency of mice within 5 min was calculated as 300 s.

[0259] 2.7 Related staff notification

[0260] When purchasing animals, inform the animal room, and when the animals have abnormal conditions, inform the pathology room for processing.

[0261] 2.8 Instrument system

[0262] DT2000 electronic balance, Changshu Shuangjie Testing Instrument Factory.

[0263] SL-2001N electronic balance, Shanghai Minqiao Precision Scientific Instrument Co., Ltd.

[0264] AL204 Mettler precision analytical balance, Mettler-Toledo (Shanghai) Co., Ltd.

[0265] Stopwatch, Shenzhen Huibuo Industry and Trade Co., Ltd.

[0266] DT200 mouse platform instrument, Chengdu Tailian Technology Co., Ltd.

[0267] 2.9 Statistical method

[0268] The experimental data was analyzed and processed by SPSS11.5 statistical software, and the statistical results were represented by mean ± standard deviation (). Firstly, normality test was performed, for data conforming to normal distribution, mean comparison was performed by one-way ANOVA, if variance was equal, pairwise comparison was performed by least significant difference method (LSD), if variance was not equal, Dunnett's T3 test was used for pairwise comparison; if not conforming to normal distribution, non-parametric test was used for statistical analysis.

[0269] 3 Results

[0270] As shown in Table 8, compared with the blank group, the latency of the model group was significantly shortened (P<0.05), and the error times were significantly increased (P<0.05), which showed that the model was successfully established; compared with the model group, the latency and error times of the control drug group had no significant difference (P>0.05), the latency of the RJYZ2 group was significantly prolonged (P<0.05), and the error times were significantly reduced (P<0.05), and the RJYZ1 group also showed that the latency was significantly prolonged (P<0.05), and the error times were significantly reduced (P<0.05).

[0271]

[0272] 4 Conclusion

[0273] The experimental results show that the pharmaceutical composition has a certain improvement effect on the memory consolidation disorder of the mice caused by sodium nitrite.

[0274] 5 Discussion

[0275] Learning and memory disorder is an important manifestation of senile dementia, and is one of the important brain functions indispensable for human and animal survival. Learning is the process of obtaining new behavior and experience by the nervous system in response to changes in the external environment, and memory is the retention and reproduction of experience after learning. Generally, learning and memory are divided into three stages, namely, acquisition, consolidation and reproduction. After a large amount of nitrite enters the body, normal hemoglobin is converted to methemoglobin, which loses the ability to carry oxygen, causing tissue hypoxia and nerve cell damage, which can cause memory consolidation disorder in animals.

[0276] The platform test is a commonly used behavioral method for testing learning and memory, but its accuracy is easily affected by many factors, such as experimental environment, experimental animals, non-specific interference, reward and punishment effect, etc.

[0277] In this experiment, the experience accumulated in the laboratory was used for reference, and the food intake of the animals was controlled to prevent the body weight from being too large, and the influence of the animals being difficult to get on the platform due to the large body weight was reduced, and the experimental environment was kept quiet, the indoor temperature and humidity, light were suitable and consistent, etc. The experimental results show that the latency and error times of the traditional Chinese medicine compound RJYZ have significant differences compared with the model group, and have a certain dose dependence, which shows that the traditional Chinese medicine compound RJYZ has a certain improvement effect on the memory consolidation disorder of the mice caused by sodium nitrite.

[0278] Test 4:

[0279] Effect of the pharmaceutical composition of the application on the memory acquisition disorder model of scopolamine-induced mice

[0280] ABSTRACT

[0281] [Objective] To observe the effect of the pharmaceutical composition of the present invention on the learning and memory abilities of mice with memory acquisition impairment induced by scopolamine. [Methods] 96 ICR mice, half male and half female, were randomly divided into 6 groups according to body weight: blank group, model group, positive drug group, RJYZ1 group, RJYZ2 group, and control drug group. The dosage of RJYZ1 group, RJYZ2 group, and control drug group was 4.0 g crude drug / kg × day. -1 This is equivalent to 10 times the clinical human dose. The positive control drug is huperzine A, with a maximum daily clinical dose of 450 μg / day in humans. The mouse dosage is 10 times the maximum clinical daily dose, i.e., 75 μg / kg × day. -1 Animals were administered the drug via gavage for 10 consecutive days, while the control and model groups received the corresponding volume of solvent. Jumping platform training began 1 hour after the last administration. Twenty minutes before training, all groups except the control group received an intraperitoneal injection of scopolamine hydrobromide 3 mg / kg. The control group received the corresponding volume of physiological saline. Training lasted 5 minutes, and memory performance was tested 24 hours later. The latency and number of errors during the first jump off the platform within 5 minutes were observed and recorded as memory indicators.

Results

Conclusion

[0282] Experimental Objective

[0283] Using a scopolamine-induced mouse model of memory acquisition impairment, the effect of the pharmaceutical composition of the present invention on improving the learning and memory abilities of mice with memory acquisition impairment was observed.

[0284] 1. Experimental Materials

[0285] 1.1 Test sample (same as the test sample in Test 1)

[0286] 1.2 Positive testing agents, tool reagents, and main reagents

[0287] Haboin (Huperzine A Tablets): Henan Tailong Pharmaceutical Co., Ltd., Batch No.: 140902.

[0288] Scopolamine hydrobromide injection: Shanghai Hefeng Pharmaceutical Co., Ltd., batch number: 02140801.

[0289] Picric acid: Taishan Yueqiao Reagent Plastics Co., Ltd., batch number: 20131001.

[0290] 1.3 Experimental system

[0291] 1.3.1 Animal strain: ICR mice.

[0292] 1.3.2 Animal level: SPF level.

[0293] 1.3.3 Animal gender and number: half male and half female, a total of 96.

[0294] 1.3.4 Animal age: 4 weeks old.

[0295] 1.3.5 Animal weight: 16-20g.

[0296] 1.3.6 Animal source: purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., qualified certificate number: 11400700098398, license number: SCXK (Jing) 2012-0001, received on June 3, 2015.

[0297] 1.3.7 Raising conditions: The animals were raised in the New Drug Evaluation Center of Hebei Province of Integrative Medicine Research Institute. The mice were raised in cages, with 12 hours of light per day, temperature 20-26℃, relative humidity 40-70%.

[0298] 1.3.8 Quarantine process: New animal quarantine period for 5 days, during the quarantine period, the animals had normal drinking and eating, good health, no disease and death signs.

[0299] 1.3.9 Feed: experimental level granular mouse feed, provided by Jiangsu Province of Collaborative Medicine Biological Engineering Co., Ltd., production license: Su Shifa (2014) 01008.

[0300] 1.3.10 Drinking water: ordinary water was filled for animals to drink freely, and the drinking water bottle was washed and changed once a day.

[0301] 1.3.11 Bedding: ordinary level bedding for experimental animals, provided by Hebei Province Experimental Animal Center, production license: SCXK (Ji) 2013-2-001.

[0302] 1.3.12 Identification: 5% picric acid marking.

[0303] 2 Experimental method

[0304] 2.1 Experimental design basis

[0305] 2.1.1 Standard adopted: The Research Guide for New Drugs of Traditional Chinese Medicine (Pharmacy, Pharmacology, Toxicology) issued by the Drug Administration Bureau of the Ministry of Public Health of the People's Republic of China, Preclinical Research Guidelines for New Drugs (Western Medicine) Compilation (Pharmacy, Pharmacology, Toxicology), "Methodology of Traditional Chinese Medicine Pharmacological Research" and "Pharmacological Experimental Methodology" published by People's Medical Publishing House, and relevant literature materials.

[0306] 2.1.2 The information provided by the entrusting unit: the proposed clinical dose of RJYZ is 24.0 g crude drug per day, and the proposed clinical dose for a 60 kg human is 0.4 g crude drug per kg. The content of the dry powder of the test sample is 5.0 g crude drug per g dry powder, and the batch number is 20150406.

[0307] 2.2 Dose and grouping

[0308] The mice were randomly divided into 6 groups according to body weight, namely, a blank group, a model group, a positive drug (huperzine A) group, an RJYZ1 group, an RJYZ2 group, and a control drug group, with 16 mice in each group. The RJYZ1 group, the RJYZ2 group, and the control drug group were designed to be 4.0 g crude drug per kg per day, which is 10 times the clinical human dose. The maximum daily dose of huperzine A in clinical use (9 tablets) is 450 μg per person per day, and the mouse dose is 10 times the maximum daily dose in clinical use, i.e. 75 μg per kg per day. See Table 9. -1 -1

[0309]

[0310] 2.3 Administration method

[0311] Gavage, consistent with the recommended oral route in clinical practice.

[0312] 2.4 Preparation and storage of test sample

[0313] Each test drug was prepared into the experimental concentration with distilled water (see Table 9), and stored at 2-8°C after preparation for standby use. The positive drug was prepared and used immediately.

[0314] 2.5 Administration of test sample

[0315] The animals were administered by gavage at 10 ml / kg, and the blank group and the model group were given distilled water, once a day for 10 consecutive days.

[0316] 2.6 Steps and observation indexes

[0317] ​​The mouse platform device is six rectangular reflection boxes, with a size of 32cm * 22.5cm * 33cm, separated into 6 rooms by black plastic plates, with a stainless steel grid at the bottom, with a grid spacing of 0.5cm, and a rubber platform with a height of 4.5cm and a diameter of 4.5cm placed at the left rear corner of each room. The mouse is placed on the platform facing the corner, and adapts to the environment for 3 minutes, then a 36V current is applied, and if the animal jumps off the platform, it will be shocked. Its normal response should be to jump back onto the platform to avoid the harmful stimulus, and most animals may jump onto the grid again or multiple times to escape the shock. When jumping off, the mouse's both feet touch the copper grid at the same time, which is considered as an error. Training for 5 minutes, 24 hours later, the memory retention test is performed, and the latency period and the number of errors of the first jump off the platform within 5 minutes are recorded.

[0318] The newly received animal is labeled, quarantined for 5 days, and after quarantine, the animals are randomly divided into 6 groups according to body weight, as described above. Intragastrically administered according to body weight, once a day, for 10 consecutive days, the blank group and the model group are given distilled water, 1 hour after the last administration, the platform training begins, 20 minutes before training, all groups except the blank group are injected with hyoscine hydrobromide 3mg / kg intraperitoneally, the blank group is given the corresponding volume of normal saline, training for 5 minutes, 24 hours later, the memory performance is tested, and the latency period and the number of errors of the first jump off the platform within 5 minutes are observed and recorded. The latency period of mice that do not jump off within 5 minutes is calculated as 300s.

[0319] 2.7 Related staff notification

[0320] When purchasing animals, notify the animal room, and when the animal appears abnormal, notify the pathology room for processing.

[0321] 2.8 Instrument system

[0322] DT2000 electronic balance, Changshu Shuangjie Test Instrument Factory.

[0323] SL-2001N electronic balance, Shanghai Minqiao Precision Scientific Instrument Co., Ltd.

[0324] Stopwatch, Shenzhen Huibuo Trade Co., Ltd.

[0325] DT200 mouse platform instrument, Chengdu Tailian Technology Co., Ltd.

[0326] 2.9 Statistical method

[0327] Experimental data was analyzed and processed using SPSS 11.5 statistical software, and the statistical results were expressed as mean ± standard deviation (X ± SD). )indicates that firstly normality test is carried out, data meeting normal distribution are compared by using One-Way ANOVA, if variance is equal, pairwise comparison is carried out by using LSD, if variance is not equal, pairwise comparison is carried out by using Dunnett's T3 test; if not meeting normal distribution, non-parametric test is used for statistical analysis.

[0328] 3 Results

[0329] As shown in Table 10, compared with the blank group, the latency of the model group was significantly shortened (P<0.05), and the error times were significantly increased (P<0.05), which showed that the scopolamine modeling was successful; compared with the model group, the latency and error times of the control drug group had no significant difference (P>0.05), the error times of the RJYZ2 group were significantly reduced (P<0.05), the latency of the RJYZ1 group was significantly prolonged (P<0.05), and the error times were significantly reduced (P<0.05).

[0330]

[0331] 4 Conclusion

[0332] The experimental results show that the pharmaceutical composition has a certain improvement effect on the memory acquisition disorder of mice caused by scopolamine.

[0333] 5 Discussion

[0334] Learning and memory disorder is an important manifestation of senile dementia, and is one of the important brain functions indispensable for human and animal survival. Learning is the process of acquiring new behavior and experience by the nervous system in response to changes in the external environment, and memory is the retention and reproduction of experience after learning. Generally, learning and memory are divided into three stages, namely, acquisition, consolidation and reproduction. Scopolamine is a M cholinergic receptor blocker, which can antagonize acetylcholine and can simulate the reversible memory disorder caused by insufficient secretion of acetylcholine, and is a commonly used initial screening model for the study of anti-senile dementia drugs.

[0335] The jumping platform experiment is a commonly used behavioral method for testing learning and memory, but its accuracy is easily affected by many factors, such as experimental environment, experimental animals, non-specific interference, reward and punishment effect, etc.

[0336] In this experiment, the experience accumulated in the previous stage of the laboratory was used for reference, and the food intake of the animals was controlled to prevent the body weight from being too large, to reduce the influence of difficulty in jumping due to excessive body weight, and to maintain a quiet experimental environment with suitable and consistent indoor temperature, humidity and light. RJYZ has a certain improvement effect on the memory acquisition disorder of mice caused by scopolamine.

[0337] Test conclusion

[0338] Aβ42 The oligomer-induced AD rat learning and memory disorder model experiment shows that the two groups of RJYZ reduce the water maze latency and total distance, increase the number of platform crossing and the time in the platform quadrant, reduce the activity of hippocampal choline system ACHE and the content of lipid peroxidation product MDA, and reduce the expression of Aβ, p-Tau and related apoptosis and inflammatory factors. In addition, the results of the platform test of three animal models of memory loss, namely, memory reproduction deficiency induced by ethanol, memory consolidation disorder induced by sodium nitrite and memory acquisition disorder induced by hyoscine, show that RJYZ can prolong the latency and reduce the number of errors.

[0339] Based on the above results, RJYZ has obvious improvement effect on Aβ 42 The oligomer-induced AD rat learning and memory ability has obvious improvement effect, and the effect may be related to improvement of cholinergic system function, reduction of oxidative stress damage, reduction of neural cell apoptosis and inhibition of inflammatory reaction. Meanwhile, the three animal models of memory loss, namely, memory reproduction deficiency, memory consolidation disorder and memory acquisition disorder, are used, and RJYZ has obvious effect of improving learning and memory.

[0340] Compared with the control drug group and the RJYZ group, there is no significant difference in the experimental results, and good effect is not shown. DETAILED DESCRIPTION

[0341] Example 1:

[0342] Cistanche tubulosa 400 g, Acanthopanax 800 g, Fructus Alpiniae oxyphyllae 400 g, Angelica sinensis 400 g.

[0343] 1. Take Fructus Alpiniae oxyphyllae, crush, and seal for standby;

[0344] 2. Take Cistanche tubulosa and Acanthopanax, and reflux extract with 50% ethanol for three times, 2 hours each time, and add 10 times of 50% ethanol each time. Filter the extract liquid, and reduce pressure to concentrate the filtrate to a clear extract with a relative density of 1.05-1.10 (60°C). Reserve the clear extract;

[0345] 3. Take Fructus Alpiniae oxyphyllae and Angelica sinensis, and add 10 times of water to heat and distill for 8 hours to collect volatile oil. Add water to the residue to decoct twice, 1.5 hours each time, and add 10 times of water each time. Combine the two decocting liquids with the distilled water decocting liquid, and reduce pressure to concentrate to a clear extract with a relative density of 1.05-1.10 (60°C). Continue to reduce pressure to concentrate to a thick extract with a relative density of 1.20-1.30 (60°C) together with the above clear extract;

[0346] 4. Take the thick extract above, and vacuum dry at a temperature of 110±5°C. Crush to obtain dry extract powder.

[0347] The dry extract powder and the volatile oil together constitute the active components of the pharmaceutical composition of the present application.

[0348] Example 2

[0349] Cistanche 380 parts, acanthopanax 700 parts, Fructus Schizandrae 400 parts, Angelica 500 parts.

[0350] (1) Raw material processing: break Fructus Schizandrae;

[0351] (2) Weighing: Cistanche, acanthopanax, Fructus Schizandrae, Angelica are weighed according to the formula proportion for standby;

[0352] (3) Take Cistanche and acanthopanax, reflux extraction with 60% ethanol for 3 times, 2 hours each time, add 10 times each time, filter the extract, reduce pressure and concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (60 ℃), for standby;

[0353] (4) Take Fructus Schizandrae and Angelica, add 10 times of water, heat and distill for 8 hours, collect the volatile oil; the water decoction is standby; add 10 times of water to the residue, decoct twice, 1.5 hours each time, combine the two decoctions with the water decoction after distillation, reduce pressure and concentrate to a clear paste with a relative density of 1.05-1.10, combine with the clear paste obtained in step (3), continue to reduce pressure and concentrate to a thick paste with a relative density of 1.20-1.30; dry and pulverize to obtain dry paste powder;

[0354] (5) After the volatile oil is encapsulated, dry and pulverize to fine powder;

[0355] (6) Mix the dry paste powder of step (4) with the fine powder of step (5), add excipients, and prepare into tablets.

[0356] Example 3

[0357] Cistanche 600g, acanthopanax 400g, Fructus Schizandrae 500g, Angelica 500g.

[0358] (1) Raw material processing: break Fructus Schizandrae;

[0359] (2) Weighing: Cistanche, acanthopanax, Fructus Schizandrae, Angelica are weighed according to the formula proportion for standby;

[0360] (3) Take Cistanche and acanthopanax, reflux extraction with 60% ethanol for 3 times, 2 hours each time, add 10 times each time, filter the extract, reduce pressure and concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (60 ℃), for standby;

[0361] (4) Take the Fructus Alpiniae Oxophyllae, Angelica, add 12 times the amount of water, heated distillation 10 hours, collect volatile oil; water decoction for later use; the residue plus 12 times the amount of water, decocting two times, 1.5 hours each time, two times of decoction and distillation after the water decoction combined, reduced pressure concentration to the relative density of 1.05~1.10 clear paste, with step (3) obtained clear paste combined, continue to reduce pressure concentration to the relative density of 1.20-1.30 thick paste; drying, crushing, dry paste powder;

[0362] (5) volatile oil after inclusion, drying, crushing into fine powder;

[0363] (6) the dry paste powder of step (4) and step (5) fine powder mixed, adding appropriate excipients, prepared into capsules.

[0364] Example 4:

[0365] Cistanche 500g, acanthopanax 600 parts, Fructus Alpiniae Oxophyllae 500 parts, Angelica 500 parts.

[0366] (1) raw material processing: take Fructus Alpiniae Oxophyllae broken;

[0367] (2) weighing: according to the formula proportion of Cistanche, acanthopanax, Fructus Alpiniae Oxophyllae, Angelica, ready for use;

[0368] (3) take Cistanche, acanthopanax, reflux extraction with 80% ethanol 2 times, 1 hour each time, add 10 times each time, filter the extract, filter the filtrate, reduce pressure concentration to the relative density of 1.05~1.10 (60 ℃) clear paste, ready for use;

[0369] (4) take Fructus Alpiniae Oxophyllae, Angelica, add 8 times the amount of water, heated distillation 9 hours, collect volatile oil; water decoction for later use; the residue plus 8 times the amount of water, decocting two times, 1.5 hours each time, two times of decoction and distillation after the water decoction combined, reduced pressure concentration to the relative density of 1.05~1.10 clear paste, with step (3) obtained clear paste combined, continue to reduce pressure concentration to the relative density of 1.20-1.30 thick paste; drying, crushing, dry paste powder;

[0370] (5) volatile oil after inclusion, drying, crushing into fine powder;

[0371] (6) the dry paste powder of step (4) and step (5) fine powder mixed, adding excipients, prepared into pills.

[0372] Control drug group:

[0373] Acanthopanax 800g; ginseng 267g; Fructus Alpiniae Oxophyllae 267g; Schisandra 178g

[0374] (1) raw material processing: take acanthopanax, Fructus Alpiniae Oxophyllae, Schisandra decoction piece, wash and reserve; ginseng is crushed, through 80 mesh sieve,60 Co irradiation sterilization, irradiation dose 3kGy, standby;

[0375] (2) Weighing: according to the formula proportion of the powder of acanthopanax senticosus, fructus schisandrae chinensis and ginseng, standby;

[0376] (3) Extraction: the weighed acanthopanax senticosus, fructus schisandrae chinensis and ginseng were extracted with water for 3 times, 1 hour each time, 9 times water was added in the first time, 8 times water was added in the second and third times, the extract was filtered and combined, standby;

[0377] (4) Concentration: the extract was concentrated under reduced pressure, concentrated to clear paste with relative density 1.05-1.10 at 60℃;

[0378] (5) Vacuum drying: vacuum oven drying was adopted, dry paste was collected, standby;

[0379] (6) Dry paste grinding: the dry paste was ground, fine powder was collected, standby;

[0380] (7) The dry paste powder and ginseng sterilization powder were mixed uniformly, dry granulation was adopted, the whole mixture was filled into capsules, and the capsules were obtained.

Claims

1. A pharmaceutical composition for treating Alzheimer's disease, characterized in that... This composition is made from the following parts by weight of active pharmaceutical ingredient: 200-600 parts of Cistanche deserticola, 400-1200 parts of Acanthopanax senticosus, 200-600 parts of Alpinia oxyphylla, and 200-600 parts of Angelica sinensis.

2. The composition according to claim 1, characterized in that This composition is made from the following parts by weight of active pharmaceutical ingredient: 300-500 parts of Cistanche deserticola, 600-1000 parts of Acanthopanax senticosus, 300-500 parts of Alpinia oxyphylla, and 300-500 parts of Angelica sinensis.

3. The composition according to claim 1, characterized in that This composition is made from the following parts by weight of active pharmaceutical ingredient: 400 parts of Cistanche deserticola, 800 parts of Acanthopanax senticosus, 400 parts of Alpinia oxyphylla, and 400 parts of Angelica sinensis.

4. The composition according to claim 1, characterized in that This composition is made from the following parts by weight of active pharmaceutical ingredient: 380 parts of Cistanche deserticola, 700 parts of Acanthopanax senticosus, 400 parts of Alpinia oxyphylla, and 500 parts of Angelica sinensis.

5. The composition according to claim 1, characterized in that... This composition is made from the following parts by weight of active pharmaceutical ingredient: 600 parts of Cistanche deserticola, 400 parts of Acanthopanax senticosus, 500 parts of Alpinia oxyphylla, and 500 parts of Angelica sinensis.

6. The composition according to any one of claims 1-5, characterized in that The preparation of its active components includes the following steps: (1) Raw material processing: Take Alpinia oxyphylla kernels and crush them; (2) Weighing: Weigh out the following ingredients according to the formula ratio: Cistanche deserticola, Acanthopanax senticosus, Alpinia oxyphylla, and Angelica sinensis. (3) Take Cistanche deserticola and Acanthopanax senticosus, and extract them by reflux with 50%-80% ethanol 2-3 times, each time for 1-3 hours, and add 8-10 times the amount each time. Filter the extract and concentrate the filtrate under reduced pressure at 60℃ to a relative density of 1.05-1.10 for later use. (4) Take Alpinia oxyphylla and Angelica sinensis, add 8-12 times the amount of water, heat and distill for 6-10 hours, and collect the volatile oil; keep the decoction for later use; add 8-12 times the amount of water to the dregs, decoct twice, each time for 1.5 hours, combine the two decoctions with the distilled decoction, and concentrate under reduced pressure to a clear extract with a relative density of 1.05-1.10, combine with the clear extract obtained in step (3), and continue to concentrate under reduced pressure to a thick extract with a relative density of 1.20-1.30; (5) Take the thick extract obtained in step (4), dry it, and pulverize it. The resulting dry extract powder and volatile oil together constitute the active component of the composition.

7. The composition according to any one of claims 1-5, characterized in that Cistanche deserticola is also known as tubular Cistanche deserticola.

8. The composition according to any one of claims 1-5, characterized in that The composition can be prepared into dosage forms such as capsules, tablets, pills, powders, or ointments.

9. The composition according to claim 8, characterized in that The tablet preparation method is as follows: (1) Raw material processing: Take Alpinia oxyphylla kernels and crush them; (2) Weighing: Weigh out the following ingredients according to the formula ratio: Cistanche deserticola, Acanthopanax senticosus, Alpinia oxyphylla, and Angelica sinensis. (3) Take Cistanche deserticola and Acanthopanax senticosus, and extract them by reflux with 50%-80% ethanol 2-3 times, each time for 1-3 hours, and add 8-10 times the amount each time. Filter the extract and concentrate the filtrate under reduced pressure at 60℃ to a relative density of 1.05-1.10 for later use. (4) Take Alpinia oxyphylla and Angelica sinensis, add 8-12 times the amount of water, heat and distill for 6-10 hours, and collect the volatile oil; set aside the decoction; add 8-12 times the amount of water to the dregs, decoct twice, each time for 1.5 hours, combine the two decoctions with the distilled decoction, concentrate under reduced pressure to a clear extract with a relative density of 1.05-1.10, combine with the clear extract obtained in step (3), and continue to concentrate under reduced pressure to a thick extract with a relative density of 1.20-1.30; dry, pulverize, and obtain dry extract powder; (5) After the volatile oil is encapsulated, it is dried and pulverized into fine powder; (6) Mix the dry powder from step (4) with the fine powder from step (5), add excipients, and prepare tablets.

10. The composition according to any one of claims 1-5, characterized in that... The application of this composition in medications for improving memory disorders.

11. The composition according to any one of claims 1-5, characterized in that The composition is used in drugs that improve cholinergic system function, reduce oxidative stress damage, reduce neuronal apoptosis, or inhibit inflammatory responses.

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