Xanthoceras sorbifolia bunge oil dha composition and its use in improving memory and tic disorders

Abstract

The application provides a Xunguan fruit oil DHA composition and application thereof in improving memory and tic disorders, and belongs to the fields of medicine and health care products; the Xunguan fruit oil DHA composition is composed of 10-90 parts by weight of Xunguan fruit oil and 10-90 parts by weight of DHA. The Xunguan fruit oil and DHA are compounded in the application, and on the basis of independent use of the Xunguan fruit oil and the DHA, the synergistic effect of the Xunguan fruit oil and the DHA significantly improves the effect of the Xunguan fruit oil and the DHA in improving memory and tic disorders; compared with the prior art, the Xunguan fruit oil DHA composition formula provided by the application is more concise, and can effectively improve memory and tic disorders.

Description

Technical Field

[0001] This invention belongs to the field of medicine and health products, and relates to a *Sapindus mukorossi* oil DHA composition and its application in improving memory and tic disorders. Background Technology

[0002] The fruit of the Chinese tallow tree has anti-inflammatory, memory-improving, cardiovascular-improving, antiviral, anticancer, and anti-HIV activities. It has certain therapeutic effects on rheumatoid arthritis, swelling and pain relief, skin fever, enuresis, Alzheimer's disease, hepatitis, and toxic liver damage. It can lower cholesterol and blood lipids. It also has a high inhibitory activity against six types of cancer cells.

[0003] The seeds of *Xanthoceras sorbifolium* contain approximately 35% oil, while the kernels contain over 60%. Nutritionally, *Xanthoceras sorbifolium* oil is an edible vegetable oil rich in unsaturated fatty acids. Besides oleic acid and linoleic acid, it also contains a rare functional component, nervonic acid, at a content of about 3%. However, the history of artificial cultivation of *Xanthoceras sorbifolium* is relatively short, and historical records of its use in folk medicine are scattered and incomplete. Furthermore, the lack of large-scale artificial cultivation has resulted in limited availability, leading to high prices and hindering its widespread use.

[0004] CN1972702B discloses the composition, function, and application of extracts of *Xanthoceras sorbifolia* and compounds isolated from the extracts, as well as methods for their preparation. It provides methods and processes for isolating, purifying, and identifying biologically active compounds from *Xanthoceras sorbifolia*. The patent also provides compositions containing compounds purified from *Xanthoceras sorbifolia*. The compounds purified from *Xanthoceras sorbifolia* contain saponin compounds. The compositions can be used for anti-cancer purposes, preventing brain aging, enhancing memory, improving brain function, treating nocturia, frequent urination, urgency, intellectual disability, dementia and Alzheimer's disease, autism, brain trauma, Parkinson's disease, and other conditions caused by brain dysfunction or impairment. They can also be used to treat arthritis, rheumatism, poor circulation, arteriosclerosis, Raynaud's phenomenon, angina pectoris, cardiac dysfunction, coronary heart disease, headache, dizziness, renal dysfunction, impotence, and premature ejaculation. However, it mainly focuses on the effects of the *Xanthoceras sorbifolia* extract, lacking research on the combination of the extract with other components, thus failing to fully utilize the efficacy of *Xanthoceras sorbifolia* oil.

[0005] Huang Yuguang et al. mentioned in their research (nutrition and comprehensive processing of Xanthoceras sorbifolium [J]. Food Research and Development, 2004, 25(3):73-76.) that Xanthoceras sorbifolium oil can be fortified with vitamin E at 0.01% by weight and used as a flavoring oil; however, they did not carry out compounding of effective components to give full play to the role of Xanthoceras sorbifolium oil in improving memory.

[0006] CN110038018A discloses a method for preparing a neurotrophic composition, relating to the field of lipid synthesis. This invention uses fatty acid triglycerides, conjugated linoleic acid glycerides, and phytosterols as raw materials, and involves transesterification reactions with 435 lipase, 1M lipase, and DF15 lipase. After cooling, decolorization, deodorization, distillation, and fine filtration, the mixture is emulsified, encapsulated, and spray-dried to obtain a dry powder. The dry powder is then mixed with phosphatidylserine, N-acetylneuraminic acid, calcium β-hydroxy-β-methylbutyrate, konjac glucomannan, mangosteen extract, elderberry, and yeast β-glucan, and homogenized to obtain a medium- to long-chain lipid composition. This process is simple, convenient, and inexpensive, and can prepare precise nutrition suitable for clinical rehabilitation of diseases such as stroke, Alzheimer's disease, depression, Parkinson's disease, and epilepsy. Although it effectively alleviates nerve damage caused by brain injury and stroke, the use of multiple raw materials and the relatively complex process pose challenges to the production and application of the composition.

[0007] Therefore, how to provide a composition that fully utilizes the role of Xanthoceras sorbifolium oil in improving memory through compounding, while ensuring that the composition is relatively simple, is one of the important research issues in this field. Summary of the Invention

[0008] This invention addresses the problem that existing technologies have limited variety and complex formulations of Xanthoceras sorbifolium oil-DHA compositions, which prevent Xanthoceras sorbifolium oil from fully exerting its effect on improving memory. It provides a Xanthoceras sorbifolium composition that, through the combination with DHA, exhibits excellent memory-improving effects. Furthermore, it has been found that the composition provided by this invention can significantly improve tic disorders, demonstrating significantly better results than using Xanthoceras sorbifolium oil and DHA independently.

[0009] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0010] A *Sapindus mukorossi* oil DHA composition, comprising the following components in parts by weight:

[0011] 10-90 parts of Sinica fruit oil and 10-90 parts of DHA.

[0012] Preferably, the *Sapindus mukorossi* oil DHA composition comprises the following components in parts by weight:

[0013] 40-60 parts of Sinica fruit oil and 40-60 parts of DHA.

[0014] More preferably, the *Sapindus mukorossi* oil DHA composition comprises the following components in parts by weight:

[0015] 50-55 parts of Xanthoceras sorbifolium oil and 45-55 parts of DHA.

[0016] Most preferably, the *Sapindus mukorossi* oil DHA composition comprises the following components in parts by weight:

[0017] 50 parts of Sinica fruit oil and 50 parts of DHA.

[0018] Preferably, the *Xanthoceras sorbifolium* oil comprises an oil product extracted from *Xanthoceras sorbifolium* kernels by mechanical extraction, leaching, aqueous extraction, ultrasonic-assisted extraction, supercritical CO2 extraction, or hydroenzymatic extraction.

[0019] Preferably, the DHA dosage is calculated based on pure DHA, and the DHA is in the form of an edible product with a DHA mass content greater than 50%.

[0020] The present invention also provides the application of the above-mentioned Xanthoceras sorbifolium oil DHA composition in the preparation of health products with the function of assisting in improving memory.

[0021] The present invention also provides the application of the above-mentioned Xanthoceras sorbifolium oil DHA composition in the preparation of a drug with the effect of improving memory and tic disorders.

[0022] The present invention also provides a health product with the function of assisting in improving memory, comprising the above-mentioned Sapindus mukorossi oil DHA composition.

[0023] The present invention also provides a medicine that improves memory and tic disorders, comprising the above-mentioned Xanthoceras sorbifolium oil DHA composition.

[0024] Preferably, the drug having the effect of improving memory and tic disorders further comprises pharmaceutically acceptable excipients.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] This invention combines *Xanthoceras sorbifolium* oil with DHA, significantly improving the effects of *Xanthoceras sorbifolium* oil and DHA on improving memory and tic disorders, compared to existing technologies. The *Xanthoceras sorbifolium* oil and DHA composition provided by this invention is simpler and can effectively improve memory and tic disorders. Attached Figure Description

[0027] Figure 1 The results show the dwell time in the central region of the rats in Experiment Example 1;

[0028] Figure 1 middle: **** P < 0.0001 vs CON; # P<0.05 vsMOD.

[0029] Figure 2 This is a rating chart of stereotyped behaviors in rats in Experiment Example 2;

[0030] Figure 3 This is a rating chart of the motor behavior of the rats in Experiment Example 2;

[0031] Figure 2 and Figure 3 middle: ** P<0.01 vs. control group; # P<0.05 vs. model group; ## P<0.01 vs. model group; @ P<0.05 vs DHA group; & P<0.05 vs. Sapindus mukorossi oil group. Detailed Implementation

[0032] It is worth noting that the raw materials used in this invention are all commercially available products, and their sources are not specifically limited.

[0033] The following sources of raw materials are provided as examples:

[0034] DHA (Batch No.: 0523003, Shandong Yuwang Pharmaceutical Co., Ltd.)

[0035] Sapindus mukorossi oil (batch number: 20220701, Shenzhen Jizhi Bencao Biomedical Co., Ltd.)

[0036] Scopolamine hydrobromide (batch number: J2129103, Aladdin Company)

[0037] Donepezil hydrochloride (batch number: 02230150, Zhien Biotechnology Co., Ltd.)

[0038] 0.9% Sodium Chloride Injection (Batch No.: 220818501, Guangdong Daxing Pharmaceutical Co., Ltd.)

[0039] Iminodiacetonitrile (IDPN, Shanghai Maclean Biochemical Technology Co., Ltd.)

[0040] Example 1: A DHA composition from Sapindus mukorossi oil

[0041] A composition was prepared by directly mixing 50 parts of *Sapindus mukorossi* oil and 50 parts of DHA.

[0042] Example 2: A Sinica oil DHA composition

[0043] A composition was prepared by directly mixing 90 parts of *Sapindus mukorossi* oil and 10 parts of DHA.

[0044] Example 3: A Sinica fruit oil DHA composition

[0045] A composition was prepared by directly mixing 10 parts of *Sapindus mukorossi* oil and 90 parts of DHA.

[0046] Experiment Example 1: Experiment on the Effect of Improving Learning and Memory in Rats

[0047] 1.1 Laboratory Animals

[0048] Sixty-four healthy male SPF-grade SD rats aged 19-21 days (40-60g) were purchased from the Guangdong Provincial Medical Laboratory Animal Center. The animals were housed at 18-23℃ with a relative humidity of 45%-55%, with free access to food and water, and were allowed to acclimatize for three days.

[0049] 1.2 Experimental Apparatus

[0050] Morris Water Maze System (Guangzhou Bit Biotechnology Co., Ltd.)

[0051] Open Field Experimental System (Anhui Zhenghua Bio-equipment Co., Ltd.)

[0052] Data was analyzed using video tracking software (SMART 3.0).

[0053] 2 Experimental Methods

[0054] 2.1 Animal grouping and administration

[0055] Sixty-four SPF-grade male rats (40-60g) were acclimatized for three days and then randomly divided into six groups according to body weight: a blank control group (CON), a model group (MOD), a positive control group (DNPQ), a group containing *Xanthoceras sorbifolium* oil (WGG), a group containing DHA, and groups 1 (DW1), 2 (DW2), and 3 (DW3), with eight rats in each group. The CON and MOD groups were administered an equivalent volume of distilled water by gavage. The DNPQ group received donepezil at 0.52 mg / kg / day. The WGG and DHA groups received *Xanthoceras sorbifolium* oil and DHA at 31.5 mg / kg / day, respectively. The DW1, DW2, and DW3 groups received the corresponding *Xanthoceras sorbifolium* oil and DHA combination at 31.5 mg / kg / day. Administered the drugs at fixed times daily for 30 consecutive days. After 18 days of administration, the model for improved learning and memory was established. Thirty minutes after daily administration, rats in all groups except the CON group were intraperitoneally injected with scopolamine 3 mg / kg / day (prepared with physiological saline). Rats in the CON group were injected with an equal volume of physiological saline. The model was established for 12 consecutive days. Behavioral experiments were started 30 minutes after model establishment, and the results of the behavioral experiments were used to evaluate whether the model was successfully replicated.

[0056] 2.2 Behavioral testing

[0057] Behavioral tests included the water maze and open field test to assess the rats' learning and memory abilities.

[0058] 2.2.1 Morris Water Maze Experiment

[0059] The Morris water maze was used to assess the learning and memory functions of rats. The water maze provides numerous indicators, with the most commonly used being escape latency in the navigation test and the number of platform crossings in the spatial exploration test. The Morris experimental setup consisted of a cylindrical pool with a diameter of 150 cm and a height of 50 cm, divided into four equal quadrants. A platform was placed in the center of one quadrant, and its position remained unchanged throughout the behavioral tests. The pool was divided into four quadrants by two perpendicular lines intersecting the center: Northwest, Northeast, Southeast, and Southwest, designated as Quadrants I, II, III, and IV, respectively. Different colored and shaped markers (circles, triangles, stars, and squares) were affixed to the inner walls of each quadrant to aid the rats' memory. The water temperature was maintained at 22±2℃ during the experiment. In the behavioral tests, the rats' swimming activities were recorded by a camera above the water maze, and the data was processed by an image acquisition device and behavioral software and stored in a computer.

[0060] Morris water maze experiment content:

[0061] A. Adaptation training: The day before the experiment, water was placed in the water maze, but no platform was placed. The rats were given water maze adaptation training for 1-2 minutes each, and then they were taken out and dried.

[0062] B. Spatial Exploration Experiment: On the 12th day of modeling, the platform was removed, and the rats were placed in the water facing the pool wall from a fixed entry point (diagonally opposite the platform). The rats explored freely for 90 seconds. The proportion of swimming time in the quadrant where the original platform was located and the number of times the rats crossed the original platform location were measured, i.e., the number of times they traversed the original platform location, reflected the rats' spatial memory ability. The more times the rats crossed the original platform location and the longer they stayed in the target quadrant, the stronger their spatial memory ability.

[0063] 2.2.2 Open Field Experiment

[0064] On the second day after modeling, an open field test was conducted. Before the test, the camera was adjusted to ensure the open field area was fully displayed on the computer program. At the start of the test, the camera program was turned on, and the observation time was set to 5 minutes. The rats were gently placed in the central area of ​​the open field, and the free movement trajectory, the distance moved within the central area, the total distance moved within the open field, the movement trajectory within the open field box, and the time spent in the peripheral and central areas were recorded. The data were analyzed using video tracking software (SMART 3.0). After each rat was tested, the plastic box was sprayed with 75% medical alcohol to thoroughly clean the box and minimize fecal odor, so as not to affect the activity of the next rat.

[0065] 2.3 Statistical Analysis

[0066] Data were analyzed using SPSS 25.0 statistical software, and all results are expressed as mean ± standard deviation. The graphs were plotted using GraphPad Prism 9.0. Independent samples t-tests were used for comparisons between groups, one-way ANOVA was used for comparisons of means across multiple groups, and LSD was used for pairwise comparisons. P < 0.05 was considered statistically significant, and P < 0.01 was considered highly statistically significant.

[0067] 3. Results of the behavioral experiment

[0068] 3.1 Results of the open field experiment

[0069] Depend on Figure 1 It can be seen that in the open field experiment, the central region time of the rats in the model group was significantly or extremely significantly different from that of the normal rats, indicating that the latency of the scopolamine-treated model group was higher than that of other groups, which shows that the learning and memory impairment model was successfully established.

[0070] Compared with the CON group, the MOD group showed a significant difference in central region time (P<0.0001); compared with the MOD group, the DW1, DW2, and DW3 groups showed a significant increase in central region time (P<0.05), and the DHA and WGG groups had lower central region times than the DW, DW2, and DW3 groups. This indicates that donepezil, *Xanthoceras sorbifolium* oil, DHA, and the DW combination all have certain effects on improving learning and memory, and the *Xanthoceras sorbifolium* oil-DHA combination is more effective in improving learning and memory in young mice than either *Xanthoceras sorbifolium* oil or DHA alone.

[0071] 3.2 Results of Space Exploration Experiments

[0072] (1) Results of space exploration experiments

[0073] Table 1 shows the frequency of platform crossings in the spatial exploration experiment for each group of rats. As can be seen from the table, the frequency of platform crossings in the CON group was concentrated at 3-4 times, while the frequency of platform crossings in the MOD group was concentrated at 0-2 times. In comparison, the number of platform crossings in the MOD group was significantly reduced, which indicates that the rats treated with scopolamine had poor learning and memory abilities, further confirming the successful establishment of the model in this experiment. Compared with the MOD group, the DNPQ group, DW1 group, DW2 group, DW3 group, WGG group, and DHA group all crossed the platform more times than the MOD group, indicating that the positive control drug, compound, *Xanthoceras sorbifolium* oil, and DHA all have the effect of improving learning and memory. The frequency of rats crossing the platform in the DW1, DW2, and DW3 groups was concentrated in 1-3 times, and in the DW2 group it even reached 4 times. The frequency of rats crossing the platform in the DNPQ group was concentrated in 2-4 times. The number of rats crossing the platform more than 2 times in the DW1, DW2, and DW3 groups was greater than that in the groups that were given DHA or Sapindus mukorossi oil alone. This indicates that the compound and donepezil both have the effect of improving learning and memory, and the compound is more effective than DHA or Sapindus mukorossi oil alone.

[0074] Table 1. Frequency Statistics of Platform Crossing Times for Each Group

[0075]

[0076] 4. Summary

[0077] This experimental case used young SD rats as subjects to observe the effect of a combination of Xanthoceras sorbifolium oil and DHA on the learning and memory of the young rats. The results showed that Xanthoceras sorbifolium oil, DHA, and the combination of Xanthoceras sorbifolium oil and DHA could improve the learning and memory of the young rats, and the combination of Xanthoceras sorbifolium oil and DHA was more effective than DHA or Xanthoceras sorbifolium oil alone in improving learning and memory.

[0078] Experiment Example 2: Efficacy Experiment in Improving Tic Disorders in Rats

[0079] 1. Laboratory animals

[0080] Fifty-six 3-week-old SPF-grade male SD rats, weighing 40-60g, were provided by the Guangdong Provincial Laboratory Animal Center. All animals were housed in an SPF-grade barrier environment animal room at a temperature of 20-25℃ and a humidity of 50%-70%.

[0081] 2. Animal grouping and model construction

[0082] After one week of acclimatization, the animals were randomly divided into 7 groups (n=8 per group): blank control group, model group, DHA group (250 mg / kg / day), *Xanthoceras sorbifolium* oil group (250 mg / kg / day), Example 1 group, Example 2 group, and Example 3 group (Examples 1-3 were administered at a 1:1 ratio, totaling 250 mg / kg / day). The gavage dose was adjusted according to the rats' body weight, administered once daily for 14 consecutive days. The corresponding drugs were administered for 7 consecutive days. Modeling began on day 8. Thirty minutes after drug administration, the blank control group received an intraperitoneal injection of 0.9% sodium chloride solution, while the other groups received an intraperitoneal injection of iminodiacetonitrile (IDPN) at a dose of 150 mg / kg, once daily for 7 days.

[0083] 3. Assessment of motor behavior and stereotyped behavior

[0084] Thirty minutes after gavage administration on day 14 (seven days after modeling), behavioral observation of the mice began. In a quiet, dark environment, the mice were removed from their rearing cages and placed in observation cages. After acclimatization, digital video recording began, with recordings every 5 minutes for 30 minutes. The mice's head and neck twitches, choreiform movements, and the number of involuntary rotations were observed. The Kadasah SA / Diamond scoring system was used to assess the mice's behavior; a score ≥1 indicated successful modeling. A double-blind method was employed to avoid subjective bias affecting the results. After testing, the excrement in the observation cages was cleaned to eliminate any influence on the next group of mice.

[0085] The scoring criteria are as follows:

[0086]

[0087] Assessment results of motor behavior and stereotyped behavior, such as Figure 2 and 3 As shown, by Figure 2 and Figure 3 It was found that, compared with the control group, the scores of motor behavior and stereotyped behavior in the model group were significantly increased (P < 0.01), indicating that the head tics model induced by IDPN in rats was successful. After 14 days of drug intervention, compared with the model group, the scores of motor behavior and stereotyped behavior in the *Xanthoceras sorbifolium* oil group, DHA group, Example 1 group, Example 2 group, and Example 3 group were significantly decreased (P < 0.05), indicating that each drug administration group could significantly improve the tics of rats. Compared with the DHA group and *Xanthoceras sorbifolium* oil group, the scores of motor behavior and stereotyped behavior in Example 1 group, Example 2 group, and Example 3 group were significantly lower in both groups (P < 0.05), indicating that there is an interaction between DHA and *Xanthoceras sorbifolium* oil, which can significantly improve tics.

[0088] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. The application of a *Sapindus mukorossi* oil DHA composition in the preparation of a drug with the effect of improving tic disorders, characterized in that: It consists of the following components in parts by weight: 40-60 parts of Sapindus mukorossi oil and 40-60 parts of DHA.

2. The application according to claim 1, characterized in that: The *Xanthoceras sorbifolium* oil includes oil products extracted from *Xanthoceras sorbifolium* kernels by mechanical methods, leaching methods, aqueous methods, ultrasonic-assisted extraction methods, supercritical CO2 extraction methods, or hydroenzymatic methods.

3. The application according to claim 1, characterized in that: The dosage of DHA is calculated based on pure DHA, and the form of DHA is an edible product with a DHA mass content greater than 50%.

4. The application according to claim 1, characterized in that: The drug also contains pharmaceutically acceptable excipients.

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