Application of betula platyphylla sap for improving memory

Abstract

The subject of the invention is an application of white birch liquid for improving memory. The invention discloses an application of a betula platyphylla sap extract in preparation of a medicine or a health food, wherein the medicine or the health food is beneficial to improving memory disorder. The invention further discloses a preparation method of the betula platyphylla sap extract, and an application of the betula platyphylla sap extract in preparation of the medicine or the health food. The white birch extract related product disclosed by the invention has the characteristics of high efficiency, safety and convenience, and can be used for effectively treating, preventing or slowing down the dysmnesia in manners of medicines, diet intervention or nutritional health-care products and the like. The invention also provides methods of treating, preventing or slowing memory disorders using the betula platyphylla extract.

Description

Technical Field

[0001] This invention relates to the use of birch sap for improving or enhancing memory. In particular, it relates to the use of a birch sap extract in the preparation of a medicine for treating, preventing, or alleviating memory impairment, or in a health food for maintaining the memory of a subject or preventing memory decline. Background Technology

[0002] As the global society ages, the incidence of degenerative brain diseases, including Parkinson's disease, Alzheimer's disease (AD), Huntington's disease, amyotrophic lateral sclerosis (ALS), and multiple sclerosis, is increasing. Memory impairment is one of the main symptoms of age-related degenerative brain diseases. Since there are currently no effective treatments for memory impairment, preventative methods such as dietary interventions or nutritional supplements are receiving increasing attention.

[0003] White birch (Betula platyphylla) belongs to the genus Betulaceae in the family Betulaceae. In the Compendium of Materia Medica, birch bark is recorded as being used to treat arthritis, skin diseases, external injuries, and for clearing heat and detoxifying. Currently, white birch is widely used in folk medicine to treat various inflammatory diseases, including hepatitis, acute tonsillitis, pneumonia, chronic bronchitis, cholecystitis, urinary tract infections, and burns. Studies have shown that white birch possesses many biological activities, such as anti-inflammatory, antioxidant, antiviral, anticancer, anti-fatigue, and neuroprotective effects. These activities are related to betulin, a triterpenoid and the main active ingredient in white birch bark.

[0004] Another product of birch is birch sap, which is typically collected between winter and spring. Fresh birch sap is a colorless, transparent liquid with a slightly sweet taste. Untreated birch sap will begin to ferment and turn sour after two or three days. Birch sap is a traditional beverage in many parts of the Northern Hemisphere. Furthermore, records indicate that birch sap also has medicinal value, used to improve liver disease, urethritis, colds, headaches, dandruff, and eczema; it can also help lower blood pressure and cholesterol, and treat scurvy, kidney disease, and gout. However, research on the effects of birch sap on improving memory impairment is not found in the background art. Therefore, there is a need for the use of birch sap extract to treat, prevent, or alleviate memory impairment.

[0005] The inventors of this application used the Morris water maze (MWM) test to evaluate the effect of oral administration of birch sap for 2 weeks on memory ability in healthy rats. The MWM test results surprisingly showed that the time and distance to find the platform were significantly shorter in the birch sap group than in the control group, indicating that birch sap helps preserve memory. This further confirms the promising potential of using birch sap extract to treat, prevent, or alleviate memory impairment, or to maintain or prevent memory decline. Summary of the Invention

[0006] This disclosure provides the use of a birch (Betula platyphylla) sap extract in the preparation of a medicament for treating, preventing, or alleviating memory impairment in a subject.

[0007] In one embodiment, the birch sap extract disclosed herein is taken from birch trees grown in Hokkaido, Japan.

[0008] In another embodiment, the birch sap is harvested in winter or spring. In yet another embodiment, the birch sap is harvested in early spring.

[0009] This disclosure also provides a method for preparing birch sap extract, which includes...

[0010] Step 1: Select a mature birch tree, drill a hole and insert a tube to guide the sap out;

[0011] Step 2: Collect the sap using a collector and store it at a low temperature;

[0012] Step 3: Filter the sap at low temperature to remove bark fragments and other impurities; and

[0013] Step 4: Using low-temperature concentration technology, most of the water in the sap is removed to obtain a high-concentration birch sap extract.

[0014] The above steps can be added or subtracted depending on the extraction of specific components or other actual needs.

[0015] This disclosure also provides the use of birch sap extract in the preparation of health foods for maintaining or preventing memory decline in subjects.

[0016] This disclosure also provides a method for treating, preventing, or alleviating memory impairment in a subject, comprising administering the birch sap extract of this disclosure to the subject.

[0017] This disclosure also provides a method for maintaining or preventing memory decline in a subject, comprising administering the birch sap extract of this disclosure to the subject.

[0018] This disclosure also provides a pharmaceutical composition for treating, preventing or alleviating memory impairment, comprising a birch sap extract and a pharmaceutically acceptable carrier and / or stabilizer.

[0019] The various objects, features, aspects and advantages of the present invention will become more apparent from the following detailed description of preferred embodiments and the accompanying drawings. Attached Figure Description

[0020] Further features and advantages of this application will become apparent from the following and more specific description of preferred embodiments, as illustrated in the accompanying drawings, wherein similar reference numerals generally refer to the same portions or elements throughout the text, and wherein:

[0021] Figure 1A-1D The chemical properties of birch sap extract (HCFO2) are as follows: Figure 1A and 1B Showing the monosaccharide composition, Figure 1C Showing molecular weight distribution, Figure 1D show 1 H NMR spectrum.

[0022] Figure 2 This is a design diagram for a rat behavior testing experiment.

[0023] Figures 3A-3C The results of the Morris water maze (MWM) test are displayed, including... Figure 3A Display the MWM trajectory diagram for each group; Figure 3B Statistical analysis showing the time it takes for rats in each group to reach the experimental platform; Figure 3C The statistical analysis shows the distances from which rats in each group reached the platform. Each group contains n = 6–10 rats.

[0024] Figure 4A and 4B The results of assessing the effect of birch sap on cerebral blood flow in rats using a laser speckle imaging system are shown. Figure 4A Typical images of cerebral blood flow in control group and experimental group rats 14 days after administration of birch sap; Figure 4B The perfusion units (perfusion units) were measured in the control and experimental groups of rats. ***, p < 0.001 compared to the control group. n = 5 rats per group. Detailed Implementation

[0025] It should be understood that the present invention is not limited to the materials, structures, procedures, methods, or constructions of the specific examples listed herein. Therefore, although some options are listed herein, those similar or having the same effect may be applied to the implementation or embodiments of the present invention, and only preferred materials and methods are described herein.

[0026] The detailed description following, in relation to the accompanying drawings, is intended to illustrate illustrative embodiments of the invention and is not intended to represent the only illustrative embodiments in which the invention may be practiced. The term "illustrative" as used throughout this specification means "serving as an example, instance, or illustration" and should not necessarily be construed as a preferred or superior embodiment. The detailed description includes specific details provided for the purpose of giving a thorough understanding of exemplary embodiments of this application. It will be apparent to those skilled in the art that exemplary embodiments of this application may be practiced without these specific details.

[0027] definition

[0028] As used herein, the following terms have the meanings described in this paragraph. It should also be understood that the terminology used in this application is for the purpose of describing specific embodiments of the invention only and is not intended to be limiting.

[0029] Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as would be understood by one of ordinary skill in the art to which this invention pertains. Generally, the nomenclature used herein is well-known and commonly used in the laboratory procedures of animal pharmacology, pharmaceutical science, separation science, and organic chemistry. In the methods described herein, actions may be performed in any order except where the timing or order of operations is explicitly stated. Furthermore, unless the express claims specify that they are performed separately, the actions referred to may be performed simultaneously. For example, the claimed action of X and the claimed action of Y may be performed simultaneously in a single operation, and the resulting method will fall within the scope of the claimed method.

[0030] Unless the context clearly indicates otherwise, the singular forms “a,” “an,” and “the” used in this specification and the appended claims include a plurality of indicated objects. For example, “an ingredient” means one ingredient or more than one ingredient.

[0031] As used herein, the term "about" will be understood by those skilled in the art and will vary to some extent depending on the context in which it is used. As used herein, when referring to measurable values ​​such as quantity or duration of time, "about" means a variation including ±20%, ±10%, ±5%, ±1%, or ±0.1% of the specific value. This is because such variations are suitable for carrying out the disclosed methods. Therefore, in some embodiments, the numerical parameters presented in this specification and the appended claims are approximate values ​​that may vary depending on the desired characteristics sought to be obtained in a particular embodiment. In some embodiments, numerical parameters should be interpreted according to the number of significant digits stated and by applying general numerical rounding techniques. The numerical values ​​presented in some embodiments of the invention may contain certain errors, which are necessarily caused by the standard deviation present in their respective experimental measurements.

[0032] As used herein, “disease” refers to a health condition of a subject who cannot maintain homeostasis and whose health will continue to deteriorate if the disease does not improve.

[0033] As used in this article, the "symptom" in the subjects refers to a state of health in which the subjects are able to maintain homeostasis, but their health is not as good as when they are free of the symptom. Without treatment, the symptom does not necessarily lead to a further deterioration in the subjects' health.

[0034] As used herein, the term "medically acceptable" means a substance, such as a carrier or diluent, that does not eliminate the biological activity or properties of the useful compounds of the present invention and is relatively non-toxic, i.e., the substance can be administered to a subject without causing undesirable biological effects or interacting with any component contained in the composition in a harmful manner.

[0035] As used herein, the term "medically effective amount," "therapeutic effective amount," or "effective amount" of a compound refers to the amount of compound sufficient to provide a beneficial effect to a subject who has been given the compound.

[0036] As used herein, the terms “subject,” “individual,” and “patient” are used interchangeably and refer to human or non-human mammals. Non-human mammals include, for example, livestock and pets such as sheep, pigs, canines, felines, and rodents. In some embodiments, the subject is a human.

[0037] As used in this article, “improvement” means the prevention, mitigation, or treatment of symptoms related to a disease or condition in a subject.

[0038] As used in this article, “prevention” means avoiding or delaying the onset of symptoms associated with a disease or condition in subjects who have not yet developed such symptoms at the time of initiation of birch sap extract administration.

[0039] As used in this article, “slowing down” means delaying the progression of symptoms in subjects who already had initial symptoms associated with the disease or condition at the time of initiation of birch sap extract.

[0040] As used in this article, the term "treatment" means reducing the frequency or severity of symptoms of a disease or condition experienced by a subject through the administration of birch sap extract.

[0041] As used in this article, the term "health food" refers to food that has substantively scientifically proven efficacy, promotes the health of test subjects, reduces the risk of disease harm, is primarily used as a dietary supplement, and is not intended for the medical purpose of treating or correcting diseases.

[0042] As used in this article, terms such as health food refer to nutritional supplements used by the subjects to replenish deficiencies, regulate bodily functions, and supplement their diet.

[0043] Birch sap

[0044] The birch sap disclosed herein is a colorless or slightly yellowish transparent liquid extracted from the bark and / or branches of birch trees. Birch sap contains betulin, polysaccharides, saponins, proteins, minerals, and other active substances, possessing high medicinal value. In Traditional Chinese Medicine theory, birch sap is classified as a heat-clearing and detoxifying medicine, and its currently disclosed therapeutic effects and benefits are as follows:

[0045] Clearing heat and detoxifying: Birch sap has the effect of clearing heat and detoxifying, which can help remove heat and dampness toxins from the body and relieve symptoms such as fever, thirst, sore throat, and rashes.

[0046] Diuretic and anti-edema effects: Birch sap can promote kidney excretion, increase urine output, help eliminate excess water in the body, and alleviate symptoms such as edema.

[0047] Anti-inflammatory and analgesic: Birch sap contains some components with anti-inflammatory and analgesic effects, such as menthol and salicylic acid, which can help relieve pain and inflammation symptoms.

[0048] Improves digestion: Birch sap can promote gastrointestinal motility, increase the secretion of digestive juices, and improve gastrointestinal problems such as indigestion and diarrhea.

[0049] Nutritional Supplements: Birch sap contains a variety of nutrients, such as vitamin C, B vitamins, iron, and potassium, which can help supplement the body's required nutrients and enhance the body's immunity and disease resistance.

[0050] In one embodiment, the birch sap is harvested annually in winter or spring, preferably in early spring. In another embodiment, the birch sap is collected from birch trees aged 20 to 50 years. In yet another embodiment, the birch trees are those growing in Hokkaido, Japan.

[0051] Dosing / Administration

[0052] Treatment regimens can affect the composition of the effective dose. Therapeutic formulations can be administered to the patient before or after the onset of disease or symptoms. Furthermore, several separate doses can be administered daily or sequentially, as well as alternating doses, or the doses can be administered continuously. Additionally, the dosage of the therapeutic formulation can be increased or decreased proportionally depending on the urgency of the treatment or prevention situation.

[0053] The birch sap extract of this disclosure can be administered to patients using known procedures, dosages, and durations to effectively treat, prevent, or alleviate memory impairment. The effective amount of birch sap extract necessary to achieve the desired therapeutic, preventative, or alleviating effect on memory impairment can vary depending on many factors, such as the activity of the specific birch sap extract used; the time of administration; the excretion rate of the extract; the duration of treatment; other drugs, compounds, or substances used in combination with the extract; the state of the patient's disease or condition, age, sex, weight, symptoms, general health status, and prior medical history; and similar factors well known in the medical field. Dosing regimens can be adjusted to provide an optimized therapeutic response. For example, several separate doses can be administered daily, or the dose can be reduced proportionally depending on, for example, the urgency of the treatment situation.

[0054] While the description of the birch sap extract provided herein primarily pertains to its suitability for ethical human administration, those skilled in the art will understand that the composition is generally suitable for administration to a wide variety of animals. Modifications to the birch sap extract are well known to be suitable for administration to a wide range of animals, and such modifications can generally be designed and implemented by a skilled veterinary pharmacologist through routine testing (if necessary). Subjects who may be administered the birch sap extract of this disclosure include, but are not limited to, humans and other primates and mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, and dogs.

[0055] Humans and animals have different tolerances to the same drug. Generally speaking, animals have a higher tolerance than humans, meaning that animals require a larger dosage per unit body weight or body surface area. The dosage per unit body weight for animals and humans can be calculated using the following general conversion table.

[0056]

[0057] Table source: International Life Sciences Institute

[0058] For example, using a rat model according to an embodiment of this disclosure, the birch sap extract of this disclosure can be administered orally at doses ranging from 3 ml / kg / time to 20 ml / kg / time. Preferably, the birch sap extract can be administered orally at doses ranging from 3 ml / kg / time to 10 ml / kg / time, for example, at doses of 6 ml / kg / time or 10 ml / kg / time. After conversion using the above-mentioned unit body weight dose conversion table, the dose of the birch sap extract of this disclosure that can be administered to humans is 0.48 ml / kg / time to 3.2 ml / kg / time, preferably 0.48 ml / kg / time to 1.6 ml / kg / time, for example, at doses of 0.48 ml / kg / time, 0.96 ml / kg / time, or 1.6 ml / kg / time. The applicable dosage of the birch sap extract disclosed herein for other animals can also be obtained using the above-mentioned conversion table per unit body weight or other conversion tables per unit body surface area. It should be noted that the above dosages are merely illustrative, and the birch sap extract disclosed herein can be administered to the subjects at an acceptable dosage range that does not produce adverse reactions.

[0059] In some embodiments, the birch sap extract of this disclosure is administered to patients at doses ranging from 1 to 5 times daily or more. In other embodiments, the birch sap extract of this disclosure may be administered to patients at doses including, but not limited to, five times daily, four times daily, three times daily, twice daily, once daily, twice daily, three times daily, four times daily, once every two days, once every three to one week, or once every two weeks. Those skilled in the art will readily understand that the frequency of administration of the various combinations of the present invention will vary depending on the subject and many factors, including but not limited to age, the disease or condition to be treated, sex, overall health status, and other factors. Therefore, the present invention should not be construed as limited to any particular dosage regimen, and the precise dosage and composition to be administered to any patient should be determined by the attending physician considering all other factors relating to the patient.

[0060] route of administration

[0061] Oral administration

[0062] Liquid formulations for oral administration may be in the form of solutions, syrups, or suspensions. These liquid formulations may be prepared by conventional methods with pharmaceutically acceptable additives, such as syrups or pharmaceutically acceptable stabilizers, preservatives, etc.

[0063] Those skilled in the art will recognize, or be able to determine, many equivalents of the particular procedures, implementations, claims, and embodiments described herein using no more than conventional experiments. Such equivalents are considered to be within the scope of this invention and are covered by the appended claims. For example, it should be understood that modifications to reaction conditions, including but not limited to reaction time, reaction size / volume, and experimental reagents (e.g., solvents, catalysts), pressure, atmospheric pressure conditions, and reducing / oxidizing agents, as well as technically recognized alternatives and the use of experiments not exceeding conventional methods, are all within the scope of this application.

[0064] It should be understood that, wherever numerical values ​​and ranges are provided herein, the descriptions in range form are merely for convenience and brevity and should not be construed as limiting the scope of the invention. Therefore, all numerical values ​​and ranges covered by the stated values ​​and ranges are included within the scope of the invention. Furthermore, all numerical values ​​falling within these ranges, as well as the upper or lower limits of the numerical ranges, are also contemplated in this application. The description of ranges should be considered as specifically disclosing all possible subranges and individual numerical values ​​within said ranges, where appropriate, including partial integers of the numerical values. For example, a description of a range from 1 to 6 should be considered as specifically disclosing subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, from 3 to 6, etc., and individual numbers within said ranges such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6, regardless of the width of the range.

[0065] The following embodiments further illustrate various aspects of the invention. However, they are not intended to limit the teachings or disclosures of the invention as described herein.

[0066] Example

[0067] The present invention will now be described with reference to the following embodiments, which are for illustrative purposes only, and the invention is not limited to these embodiments but covers all variations that may appear as a result of the teachings provided herein.

[0068] Materials and Methods

[0069] The following procedures can be used to prepare and / or test exemplary birch sap extracts of the present invention.

[0070] Birch sap extract

[0071] The following steps are used to extract sap from birch (Betula platyphylla) and prepare birch sap extract:

[0072] Step 1: Select a mature birch tree and make a cut in the bark to encourage sap flow.

[0073] Step 2: Use a collector to collect the sap and avoid external contamination.

[0074] Step 3: Filter the collected sap at room temperature to remove bark fragments and other impurities.

[0075] Step 4: Using low-temperature concentration technology, most of the water in the sap is removed to obtain a high-concentration birch sap extract.

[0076] laboratory animals

[0077] Male Sprague-Dawley rats (SD, 150-200g) were purchased from BioLASCO and housed at the Laboratory Animal Center of Fu Jen Catholic University. All animal experiments were approved by the Laboratory Animal Management Committee of Fu Jen Catholic University (Approval No.: A11018). In accordance with the 3R principle, all experiments were conducted to minimize animal suffering and use the minimum number of animals required to produce reliable results.

[0078] Statistical analysis

[0079] Statistical analysis was performed using SPSS 16.0 software. Data are expressed as mean ± standard deviation of mean (SEM). One-way analysis of variance (ANOVA) was performed, followed by Tukey post-hoc comparison test. The criterion for statistical significance was p < 0.05.

[0080] Example 1 - Birch Sap Composition Analysis

[0081] Freeze-drying process for liquid polysaccharides

[0082] 50 ml of liquid birch sap extract was placed at -80°C overnight. After the birch sap extract froze into ice, it was freeze-dried to allow the water to sublimate, ultimately forming a solid birch sap extract (HCF-02; 23.5 g).

[0083] Molecular weight analysis

[0084] HCF-02 was dissolved in distilled water to a concentration of 1 mg / mL, and analysis was performed using two sets of high-performance size-exclusion chromatography (HPSEC) columns: G4000PWXL (7.8 x 300 mm) and G3000PWXL (7.8 x 300 mm). This analysis was performed using a Viscotek TDA-3-1 relative viscometer at a flow rate maintained at 0.5 mL / min, with deionized water as the dissolution solution. Calibration curves were established using the Sodex P-82 series from Showa Denko America, with different pullulan standards, raffinose, and glucose.

[0085] Monosaccharide analysis

[0086] The monosaccharide composition of HCF-02 was determined using high-performance anion exchange chromatography (HPAEC) with a pulsed amperometric detector (PAD) featuring a gold working electrode and a Carbopac PA-10 anion exchange column (4.6 x 250 mm) (from Dionex BioLC). For analysis, 1 mg of HCF-02 was hydrolyzed in 1.95 N trifluoroacetic acid (TFA) at 80 °C for 6 hours. After hydrolysis, residual TFA was removed under reduced pressure, and the resulting sample was dissolved in Milli-Q water. The aqueous solution was filtered through a 0.22 μm filter before HPAEC analysis. The mobile phase consisted of an isocratic solution of 18 mM NaOH, maintained at ambient temperature, with a flow rate of 1.0 mL / min. Monosaccharide identification and quantification were performed by comparing the analytical results with those of standard monosaccharides. Nine monosaccharide standards were selected: inositol, sorbitol, fucose, galactosamine, glucosamine, galactose, glucose, mannose, and fructose, all obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). Data collection and integration were performed using the PRIME DAK system (HPLC Technology Ltd., UK).

[0087] The monosaccharide analysis method described above was used to analyze the birch sap extract (HCF-02). The results of the monosaccharide analysis of the birch sap are as follows: Figure 1A and Figure 1B As shown in the figure. The analysis results show that the ratio of the three main monosaccharides, fructose, glucose, and fucose, in the birch sap extract used is 27:17:10.

[0088] Analysis using single-column high-performance molecular sieve chromatography (HPSEC) showed that the molecular weight of birch sap was approximately or less than 1.29 kDa. Figure 1C ). Birch sap 1 H NMR spectroscopy ( Figure 1D The results showed two anomeric protons at δH 5.13 and 4.45. The signals at δH 5.13 and δH 4.45 exhibited a bimodal distribution with a ratio of 100:124, corresponding to two monosaccharide residues and an α-anomeric conformation, respectively. 3 J 1,2 =3.4Hz) and β-anodic configuration ( 3 J 1,2=7.7Hz). Based on the above analysis results, the amount of birch sap extract used in subsequent biological experiments can be further quantified.

[0089] Example 2 - Morris Water Maze (MWM) Test

[0090] Thirty male Sprague-Dawley rats weighing 160-200g were used. The rats were housed in plastic cages, fed pelleted food, and had free access to drinking water. The room temperature was controlled at 22±2℃, with a 12-hour light:12-hour dark cycle. After 3 days of MWM training, the rats were divided into experimental and control groups. The experimental group received a birch sap extract solution orally daily, while the control group received an equal volume of 0.9% saline solution orally for two weeks. Two weeks later, behavioral tests were conducted using the MWM video analysis system (e.g., Figure 2 (As shown).

[0091] Control group:

[0092] Oral saline solution, 14 days

[0093] Experimental group:

[0094] Low-dose birch sap extract group: Oral administration of birch sap extract 3ml / kg for 14 days.

[0095] Medium-dose group of birch sap extract: Oral administration of 6 ml / kg of birch sap extract for 14 days.

[0096] High-dose birch sap extract group: Oral administration of birch sap extract 10ml / kg for 14 days.

[0097] The MWM test is designed to assess performance in spatial learning and memory. A circular water tank with a diameter of 55 cm and a height of 25 cm is used, filled to a depth of 20 cm with a solution at 25 ± 1 °C. The water tank is divided into four quadrants, with all test platforms placed in the center of a fixed quadrant.

[0098] Days 0-3 are the training phase, with four training sessions per day. The pool is filled with clean, clear water, and the experimental platform is slightly above the water surface. First, the rats are placed on the platform to familiarize them with its presence in the pool. Then, the rats are gently lowered into the pool, facing the pool wall. The time it takes for each rat to reach the platform is measured at 120 seconds. After reaching the platform, the rat stays for 15 seconds. If a rat fails to find the platform within the set time, the operator gently guides it to the platform and it stays there for 30 seconds.

[0099] After two weeks of oral administration of physiological saline to the control group and birch sap extract to the three experimental groups, rats entered the experimental phase. At this time, the pool was filled with water so that the experimental platform was slightly below the water surface, and the water was colored with a non-toxic pigment to make the platform invisible from the water's surface. Similar to the training phase, rats were gently lowered into the pool facing the pool wall. The time it took for each rat to reach the platform was measured at 120 seconds. If a rat reached the platform, it was allowed to stay there for 15 seconds. Rats that failed to find the platform were guided to it and allowed to stay for 30 seconds. The time for the failed rats was recorded within 120 seconds. A video tracking system (version 1.17, SINGA Technology Corporation) was used to record the swimming path from the point of descent into the pool to the hidden platform, the escape latency, and the distance traveled within the covered area.

[0100] To determine the effects of birch sap on spatial learning and memory, the performance of rats in the MWM test was analyzed, for example... Figure 3A The swimming path diagram is shown. In the MWM test, rats in the experimental groups fed different amounts of birch sap found the platform in a shorter time than rats in the control group (p<0.005, reference). Figure 3B Similarly, rats in the experimental groups fed different amounts of birch sap swam significantly shorter distances than the control group (p<0.001; reference). Figure 3C These results show that the experimental group of rats fed birch sap had better memory retention than the control group of rats not fed birch sap.

[0101] Example 3 - Effect of birch sap on cerebral blood flow in rats

[0102] Cerebral blood flow is related to memory function. To further verify the effect of birch sap on memory preservation, high-dose rats were given 10 ml / kg of birch sap orally for 14 days, and whole-brain cerebral blood flow was assessed using a laser speckle imaging system.

[0103] Monitoring cerebral blood flow using a laser speckle imaging system

[0104] Cerebral blood flow in rats was monitored using a laser speckle imaging system (RFLSI III, RWD) (Zhang, Y. et al., 1alpha,25-Dihydroxyvitamin D3 Promotes Angiogenesis After Cerebral Ischemia Injury in Rats by Upregulating the TGF-beta / Smad2 / 3 Signaling Pathway. Front Cardiovasc Med 2022,9,769717; Lu CW et al., Gypenoside XVII Reduces Synaptic Glutamate Release and Protects against Excitotoxic Injury in Rats. Biomolecules. 2024 May 16;14(5):589). In short, rats (n=5 per group) were anesthetized with 3% isoflurane and placed prone in a stereotactic head frame. A midline incision was then made in the skull to expose the top of the skull. During image acquisition, the exposed area was kept clean and dry using cotton balls. Subsequently, the skull was irradiated with a laser, and high-resolution speckle images of blood flow were recorded using a CMSO camera. The laser speckle imaging system displayed the actual image and the speckle image. Cerebral blood flow was continuously measured for 15 seconds under the following settings: observation height 25 cm; laser irradiation area 1.7 cm; image matrix 2064 × 1544. Cerebral blood flow data are expressed in perfusion units.

[0105] The results are as follows Figure 4A and 4B As shown, the cerebral blood flow in the experimental group of rats orally administered birch sap was significantly higher than that in the control group (p<0.001). Cerebral blood flow homeostasis is essential for the normal functioning of nerve cells, playing a crucial role in memory maintenance. The above animal experimental results confirm that the birch sap extract disclosed herein has a significant therapeutic, preventative, or alleviating effect on memory impairment, demonstrating its therapeutic, ameliorative, or preventative effects against degenerative brain diseases. Therefore, birch sap shows promise for the development of drugs, health foods, and / or related products for degenerative brain diseases.

[0106] List of implementation methods:

[0107] The following illustrative implementations are provided, and their numbers should not be interpreted as indicating a degree of importance.

[0108] Implementation method 1 provides the use of birch sap extract in the preparation of a medicament for treating, preventing or alleviating memory impairment in a subject.

[0109] Embodiment 2 provides the use described in Embodiment 1, wherein the birch sap extract is taken from birch trees grown in Hokkaido, Japan.

[0110] Embodiment 3 provides the use described in Embodiment 1, wherein the birch sap extract is extracted from the sap of birch trees that grow from winter to spring.

[0111] Implementation 4 provides the use described in Implementation 1, wherein the drug is used to treat, prevent or alleviate memory impairment.

[0112] Implementation 5 provides the use described in Implementation 1, wherein the memory impairment is caused by Parkinson's disease, Alzheimer's disease (AD), Huntington's disease, amyotrophic lateral sclerosis (ALS), and / or multiple sclerosis.

[0113] Implementation 6 provides the use described in Implementation 1, wherein the subject is a human.

[0114] Implementation 7 provides the use described in Implementation 1, wherein the subject is a non-human mammal.

[0115] Implementation 8 provides the use described in Implementation 1, wherein the drug is administered to the subject in a dosage range of five times daily, four times daily, three times daily, twice daily, once daily, twice daily, three times daily, four times daily, once every two days, once every three to one week, or once every two weeks.

[0116] Embodiment 9 provides the use described in Embodiment 6, wherein the drug is administered to the subject at a dose of 0.48 ml / kg / time to 3.2 ml / kg / time.

[0117] Embodiment 10 provides the use described in Embodiment 6, wherein the drug is administered to the subject at a dose of 0.48 ml / kg / time to 1.6 ml / kg / time.

[0118] Embodiment 11 provides the use described in Embodiment 6, wherein the drug is administered to the subject at a dose of 0.48 ml / kg / dose, 0.96 ml / kg / dose, or 3.2 ml / kg / dose.

[0119] Embodiment 12 provides a method for preparing birch sap extract, comprising:

[0120] Step 1: Select a mature birch tree, drill a hole and insert a tube to guide the sap out;

[0121] Step 2: Collect the sap using a collector and store it at a low temperature;

[0122] Step 3: Filter the sap at low temperature to remove bark fragments and other impurities; and

[0123] Step 4: Using low-temperature concentration technology, most of the water in the sap is removed to obtain a high-concentration birch sap extract.

[0124] Implementation 13 provides the method described in Implementation 12, wherein the birch tree is a birch tree growing in Hokkaido, Japan.

[0125] Implementation 14 provides the method of implementation 12, wherein the birch sap is harvested from winter to spring.

[0126] Implementation 15 provides the method of implementation 14, wherein the birch sap is harvested in early spring.

[0127] Implementation method 16 provides the use of birch sap extract in the preparation of health foods that maintain or prevent memory decline in subjects.

[0128] Embodiment 17 provides the use described in Embodiment 16, wherein the birch sap extract is taken from birch trees grown in Hokkaido, Japan.

[0129] Embodiment 18 provides the use described in Embodiment 16, wherein the birch sap extract is taken from birch trees that grow from winter to spring.

[0130] Implementation 19 provides the use described in Implementation 16, wherein the subject is a human.

[0131] Implementation 20 provides the use described in Implementation 16, wherein the subject is a non-human mammal.

[0132] Implementation 21 provides the use described in Implementation 18 or 19, wherein the health food is administered to the subject in a dosage range of five times a day, four times a day, three times a day, twice a day, once a day, twice a day, three times a day, four times a day, once every two days, once every three to one week, or once every two weeks.

[0133] Implementation 22 provides the use described in Implementation 18, wherein the health food is administered to the subject at a dose of 0.48 ml / kg / time to 3.2 ml / kg / time.

[0134] Embodiment 23 provides the use described in Embodiment 18, wherein the health food is administered to the subject at a dose of 0.48 ml / kg / time to 1.6 ml / kg / time.

[0135] Embodiment 24 provides the use described in Embodiment 18, wherein the health food is administered to the subject at a dose of 0.48 ml / kg / time, 0.96 ml / kg / time, or 3.2 ml / kg / time.

[0136] Embodiment 25 provides a method for treating, preventing, or alleviating memory impairment in a subject, comprising administering a birch sap extract prepared as described in any one of Embodiments 12 to 15 to the subject.

[0137] Implementation 26 provides a method for maintaining or preventing memory decline in a subject, comprising administering a birch sap extract prepared as described in any one of Implementations 12 to 15 to the subject.

[0138] Implementation 27 provides a pharmaceutical composition for treating, preventing or alleviating memory impairment, comprising a birch sap extract and a pharmaceutically acceptable carrier and / or stabilizer.

[0139] Embodiment 28 provides a solid birch sap extract, which is prepared by freeze-drying a birch sap extract prepared as in any one of Embodiments 12 to 15.

[0140] Embodiment 29 provides the solid birch sap extract of Embodiment 28, wherein the solid birch sap extract comprises inositol, sorbitol, fucose, galactosamine, glucosamine, galactose, glucose, mannose and fructose.

[0141] Embodiment 30 provides the solid birch sap extract described in Embodiment 29, wherein the ratio of fructose, glucose and fucose is 27:17:10.

[0142] While this invention has been disclosed with reference to specific embodiments, it will be apparent to those skilled in the art that other embodiments and variations can be devised without departing from the true spirit and scope of the invention. The appended claims are intended to be interpreted as encompassing all such embodiments and equivalent variations.

Claims

1. Use of birch (Betula platyphylla) sap extract in the preparation of medicines for the treatment, prevention or relief of memory impairment.

2. The use according to claim 1, wherein the birch sap extract is derived from birch trees grown in Hokkaido, Japan.

3. The use according to claim 1, wherein the birch sap extract is extracted from the sap of birch trees that grow from winter to spring.

4. The use according to claim 1, wherein the memory impairment is caused by Parkinson's disease, Alzheimer's disease (AD), Huntington's disease, amyotrophic lateral sclerosis (ALS), and / or multiple sclerosis.

5. The use according to claim 1, wherein the birch sap extract is prepared by filtration and low-temperature concentration, wherein the birch sap extract comprises inositol, sorbitol, fucose, galactosamine, glucosamine, galactose, glucose, mannose and fructose.

6. The use according to claim 1, wherein the subject is a human.

7. The use according to claim 1, wherein the subject is a non-human mammal.

8. The use according to claim 6, wherein the drug is administered to the subject at a dose of birch sap extract ranging from 0.48 ml / kg / time to 3.2 ml / kg / time.

9. A solid birch sap extract, which is prepared by freeze-drying a birch sap extract obtained by filtering and low-temperature concentration of birch sap, wherein the solid birch sap extract comprises inositol, sorbitol, fucose, galactosamine, glucosamine, galactose, glucose, mannose and fructose.

10. The solid birch sap extract according to claim 9, wherein the ratio of fructose, glucose and fucose is 27:17:10.

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