Compound chewable tablet with anti-hypoxia activity as well as preparation method and application of compound chewable tablet

Abstract

The invention discloses a compound chewable tablet with anti-hypoxia activity and application of the compound chewable tablet, and belongs to the technical field of functional food and health care products. The compound chewable tablet is prepared by compounding an olive extract, a Chinese wolfberry fruit extract, an American ginseng extract, skim milk powder, microcrystalline cellulose, mannitol, citric acid, a sweetening agent, mint powder and a small amount of tabletting auxiliary materials according to a specific weight percentage. The chewable tablet preparation is good in taste, rapid in disintegration and high in bioavailability, the SOD and NO levels of an organism can be remarkably improved, oxidative stress indexes such as MDA and LDH are reduced, and the hypoxia tolerance is enhanced. And the nutritional powder is suitable for functional nutrition intervention of plateau people, mental workers, chronic fatigue state or sub-health people. The product takes plant extracts as main active ingredients, is high in safety and mild in process, and has good industrialization and market application prospects.

Description

Technical Field

[0001] The present invention belongs to the field of functional foods and health products, and specifically relates to a compound chewable tablet with anti-hypoxia activity, which is based on olive extract and compounded with wolfberry, American ginseng and other plant ingredients, as well as a preparation method and application thereof. Background Art

[0002] Hypoxia is a common pathological and environmental stress state, widely present in scenarios such as high-altitude hypoxia, respiratory diseases, cardiovascular and cerebrovascular dysfunction, malignant tumor microenvironments, and chronic fatigue syndrome. Both acute and chronic hypoxia can lead to impaired mitochondrial energy metabolism, accumulation of reactive oxygen species, increased cell apoptosis, and multi-organ dysfunction. The oxidative stress response induced by hypoxia is considered a key pathological link in the development and progression of various diseases. Therefore, the development of safe and effective interventions with dual anti-hypoxia and antioxidant properties is of great significance for maintaining health and preventing and controlling chronic diseases.

[0003] Currently, commonly used anti-hypoxia interventions rely primarily on synthetic drugs, such as erythropoietin, glucocorticoids, and vitamin-based antioxidants. Although these drugs have demonstrated efficacy in certain clinical settings, they are generally associated with significant side effects, high risks of long-term use, and high dependency. These drugs are particularly unsuitable for long-term use in subclinical hypoxia states or in healthy individuals. Furthermore, these drugs typically have a single mechanism of action, making it difficult to achieve multi-target and multi-pathway physiological regulation, and therefore cannot meet the intervention needs of complex hypoxic states.

[0004] In recent years, natural plant extracts, especially those rich in functional active substances such as polyphenols, flavonoids, and iridoids, have shown broad application prospects in anti-hypoxia, anti-oxidation, and energy metabolism regulation due to their broad-spectrum biological activity, high safety, and good compliance. Among them, olive extract, an important medicinal and edible plant in the Mediterranean region, is rich in various active ingredients such as hydroxytyrosol, oleuropein, and squalene. It has been confirmed in multiple animal models and in vitro experiments to have multi-target biological effects such as antioxidant, anti-inflammatory, free radical scavenging, delayed cell apoptosis, regulation of the nitric oxide pathway, and improvement of mitochondrial function.

[0005] However, there is currently a lack of research on functional formulations combining olive extracts and other synergistic plant extracts (such as wolfberry and American ginseng). In particular, there is no systematic, publicly available technical solution for chewable tablets with high compliance, good taste, and portability. Furthermore, there is a lack of systematic research on the pharmacodynamics, optimal dosage range, time window of action, and anti-hypoxic mechanisms of such compositions in hypoxia models.

[0006] Therefore, there is an urgent need to develop a new type of compound functional chewable tablet based on olive and synergistic plant extracts, which can not only regulate oxidative stress and energy metabolism through multiple pathways, but also has good taste, safety and palatability. It is suitable for daily nutritional intervention for people with sub-health or chronic diseases who need to improve their tolerance to hypoxia, such as people in plateau areas, the elderly, and patients with chronic fatigue. Summary of the Invention

[0007] The present invention aims to address the shortcomings of existing anti-hypoxia intervention methods, such as a single formulation type, low bioavailability, and significant side effects. The invention provides a hypoxia-resistant functional chewable tablet made with olive extract as the core and compounded with multiple plant ingredients, including wolfberry and American ginseng. The chewable tablet is made from natural raw materials, undergoes a gentle process, effectively retains the plant's active ingredients, and possesses excellent taste, portability, and physiological functions. It is particularly suitable for daily intervention and conditioning in high-altitude environments, chronic hypoxia states, and sub-healthy populations. This fills a gap in the research and development of natural plant-derived anti-hypoxia functional food formulations and is suitable for daily health conditioning and auxiliary intervention in chronic hypoxia states.

[0008] The above-mentioned purpose of the present invention is achieved through the following technical solutions.

[0009] The present invention provides a compound chewable tablet with anti-hypoxia activity, comprising the following components in percentage by weight: olive fruit powder: 15%-25%; wolfberry extract: 15%-25%; American ginseng extract: 2%-6%; skim milk powder: 18%-28%; microcrystalline cellulose: 15%-22%; mannitol: 8%-12%; citric acid: 0.5%-1.5%; sweetener: total amount 0.2%-0.8%; mint powder: 0.1%-0.5%; and tableting excipients ≤0.5%.

[0010] The olive extract contains hydroxytyrosol, oleuropein and squalene; the wolfberry extract contains wolfberry polysaccharide; and the American ginseng extract contains ginsenosides Rb1 and Rg1.

[0011] The sweetener is one or both of stevioside and aspartame; the tableting auxiliary material is one or both of silicon dioxide and magnesium stearate.

[0012] The mass of a single chewable tablet is 0.5 to 1.2 g, preferably 0.7 g per tablet, and each tablet contains no less than 100 mg of the active ingredient of olive fruit powder.

[0013] The chewable tablets are prepared by direct compression and have a disintegration time of ≤3 minutes; The chewable tablet further comprises one or more of natural flavors, plant pigments, oral lubricants or food-grade adhesives to improve taste and appearance.

[0014] The preparation method of the compound chewable tablet comprises the following steps: (1) Mix the components in proportion and stir at 45-65 rpm for 20-40 minutes; (2) The mixed powder is dry-pressed into tablets weighing 0.5 to 1.2 g; (3) Store the tablets sealed and away from light.

[0015] The present invention also provides the use of the compound chewable tablet in the preparation of anti-hypoxia functional food or health care product, which is used to increase the levels of superoxide dismutase (SOD), nitric oxide (NO) and adenosine triphosphate (ATP) in the body; Used to reduce lactate dehydrogenase (LDH) and malondialdehyde (MDA) oxidative stress indicators.

[0016] The anti-hypoxia functional food or health product is suitable for improving and intervening in plateau hypoxia, mental fatigue, chronic fatigue syndrome or senile hypoxia.

[0017] The compound chewable tablet achieves anti-hypoxia effect by regulating oxidative stress pathways, improving mitochondrial metabolism and inhibiting cell apoptosis.

[0018] In the compound olive chewable tablets provided by the present invention, there is a clear functional synergistic mechanism between the three selected plant extracts, which is mainly reflected in the following aspects: The main active ingredient in olive extract is hydroxytyrosol, which activates the Nrf2-ARE antioxidant pathway, upregulates the expression of antioxidant enzymes such as SOD, GSH-Px, and HO-1, reduces ROS accumulation, improves mitochondrial energy metabolism, and delays hypoxia-induced oxidative damage.

[0019] Lycium barbarum extract contains high levels of Lycium barbarum polysaccharides, which can activate the Nrf2 and PI3K / Akt signaling pathways, enhance SOD and CAT activity, promote NO synthesis, and improve blood oxygen carrying capacity, exhibiting an anti-hypoxic effect in the central nervous system. This has a synergistic effect with hydroxytyrosol.

[0020] The main components of American ginseng extract are ginsenosides. Monomeric saponins such as Rg1 and Rb1 have been shown to inhibit hypoxia-induced upregulation of mTOR and HIF-1α, reducing hypoxia-induced apoptosis. Furthermore, saponins regulate NO synthase (NOS), further collaborating with these two to maintain mitochondrial stability.

[0021] Therefore, the present invention achieves multi-target synergistic control of hypoxic injury response through the bidirectional activation of the antioxidant system (Nrf2) by hydroxytyrosol + wolfberry polysaccharides and the anti-apoptosis + mTOR / AMPK regulation of American ginsenosides, avoiding the problem of redundant or weak effects of single component mechanisms. It is a functional combination of "multi-pathway, multi-target intervention".

[0022] Compared with the prior art, the present invention has the following advantages: 1. Multi-target anti-hypoxia and anti-oxidation synergistic effects (1) Synergistic effect of compound active ingredients The three plant extracts in the compound olive chewable tablets provided by this invention exhibit a clear synergistic mechanism, significantly enhancing their anti-hypoxic activity. Hydroxytyrosol and oleuropein, the main components of the olive extract, activate the Nrf2 / ARE antioxidant signaling pathway, significantly increasing SOD activity while reducing MDA levels, scavenging excess free radicals, and alleviating oxidative stress damage. Lycium barbarum polysaccharides and American ginseng saponins (primarily Rb1 and Rg1) synergistically inhibit the upregulation of the mTOR pathway, effectively reducing cell apoptosis under hypoxic conditions. This combination also lowers serum LDH levels, reflecting their protective effect on cellular structural integrity. The combination of these three exhibits a synergistic effect far exceeding that of the individual components alone. Animal model experiments showed that in the compound group: SOD activity was significantly increased, MDA levels were significantly decreased; NO levels were elevated, indicating enhanced vasodilation and oxygenation capacity; ATP levels were elevated, indicating significant improvement in energy metabolism; and LDH levels were decreased, indicating a protective effect on cell membrane integrity.

[0023] In summary, the present invention achieves synergistic intervention of multiple targets through three dimensions: anti-oxidation + energy metabolism enhancement + hypoxic damage protection, and has substantial technological progress and innovative value.

[0024] (2) Comprehensive improvement of oxidative stress indicators In an animal model of hypoxia, the compound olive chewable tablets of the present invention can effectively improve multiple physiological and biochemical indicators related to oxidative stress, showing a significant systemic protective effect.

[0025] The specific manifestations are as follows: Decreased LDH levels indicate that damage to cell membrane integrity is alleviated and cell structure is protected; Decreased MDA levels indicate that lipid peroxidation is inhibited and the degree of oxidative damage is significantly reduced; Upregulation of SOD activity: This indicates that the body's endogenous antioxidant enzyme system is activated, which can eliminate various free radicals; Increased NO levels: reflects enhanced vasodilation, which helps improve tissue oxygenation capacity; Increased ATP content: indicates that mitochondrial function has recovered and energy metabolism efficiency has improved.

[0026] The present invention can systematically alleviate cellular stress and damage responses under hypoxic conditions by regulating multiple oxidative stress-related physiological parameters, demonstrating powerful physiological regulation capabilities and application potential.

[0027] 2. Dosage form design and bioavailability advantages (1) Rapid disintegration and high absorption efficiency By compounding microcrystalline cellulose (15%-22%) with mannitol (8%-12%), the disintegration time is achieved to be ≤3 minutes (Chinese Pharmacopoeia standard); The chewable tablet form is directly absorbed through the oral mucosa, and its bioavailability is 30% higher than that of traditional capsules.

[0028] (2) Optimization of taste and compliance The addition of skimmed milk powder (18%~28%) and mint powder (0.1%~0.5%) gives the product a natural milky flavor and a refreshing feeling; stevioside adjusts the sweetness and improves palatability.

[0029] 3. Security and Industrial Value (1) Natural raw materials and low toxicity All components are food-grade raw materials, and the acute toxicity test LD50 in rats is greater than 5000 mg / kg. No abnormal liver and kidney function was found after long-term use (90 days).

[0030] (2) Process simplification and cost control The direct tableting method eliminates the wet granulation step, reducing production energy consumption by 20%; the tablet weight of 0.5-1.2g is designed to be compatible with standard tableting equipment, with a production capacity of up to 100,000 tablets per batch.

[0031] 4. Application scenario expansion (1) Multi-scenario adaptability High-altitude hypoxia: Mouse models showed a 26.4% increase in survival time (simulated environment at an altitude of 5,000 meters); Mental fatigue: In clinical trials, the cognitive function scores of subjects increased by 18% (P < 0.05).

[0032] (2) Wide coverage of the population It is suitable for everyone from plateau workers to middle-aged and elderly people, and the flexible dosage of 1 to 4 tablets per day can meet differentiated needs.

[0033] 5. Ecological and social benefits (1) Innovation in resource utilization Olive extract is derived from agricultural by-products (olive leaves), achieving high-value utilization of waste and supporting the development of characteristic plant resources in the Northwest region.

[0034] (2) Market differentiation competitiveness: There are currently no similar compound chewable tablet products on the market, filling the gap in the dosage form of hypoxia-resistant functional food. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 Effect of compound olive chewable tablets on the survival time of mice in normobaric hypoxia; Figure 2 Effects of compound olive chewable tablets on LDH activity in the blood of normobaric hypoxic mice; Figure 3 Effects of compound olive chewable tablets on SOD activity in the blood of normobaric hypoxic mice; Figure 4 Effects of compound olive chewable tablets on MDA content in the blood of normobaric hypoxic mice; Figure 5 Effects of compound olive chewable tablets on ATP content in the blood of normobaric hypoxic mice; Figure 6 Effect of compound olive chewable tablets on the NO content in the blood of normobaric hypoxic mice. DETAILED DESCRIPTION

[0036] The following specific examples further illustrate the compound olive chewable tablets with hypoxia resistance activity, their preparation method, and applications described in the present invention. It should be understood that these examples are only used to illustrate the present invention and should not limit the scope of protection of the present invention. Any equivalent replacement or improvement made within the spirit and technical solution of the present invention should be included in the scope of protection of the claims of the present invention. Example 1: Preparation of compound olive chewable tablets (1) Preparation of olive extract: Take 1000 g of dried olive fruit powder, add 10 times the volume of purified water (i.e., 10 L), heat and boil for 1.5 hours, and collect the decoction; add 8 L of purified water to the residue and boil again for 1.5 hours. After the two decoctions are combined, filter and remove the residue through a Buchner funnel to obtain a clear filtrate. The obtained filtrate is passed through a pretreated macroporous adsorption resin column (such as AB-8 or DA-201) at a flow rate of 1-2 BV / h to fully adsorb the active ingredients. After adsorption is completed, the impurities are washed with deionized water, and then eluted with (1-100)% ethanol, and the eluate is collected. The elution process can be carried out in stages, with the eluate of each stage controlled at 2-3 times the column volume, and the total elution volume is 6-10 BV. After the eluates are combined, they are concentrated under reduced pressure and spray-dried to finally obtain a light brown powdered olive extract, which mainly contains components such as hydroxytyrosol and oleuropein.

[0037] (2) Preparation of wolfberry extract: 1000 g of dried Ningxia wolfberry fruit was crushed and refluxed with 10 times the volume of purified water (i.e., 10 L) at 95°C for 2 times, each time for 90 minutes. The filtrates were combined and concentrated under reduced pressure to 1 / 4 of the original volume.

[0038] Anhydrous ethanol was added to the concentrate to adjust the alcohol concentration to 70%, and the solution was allowed to settle for 12 hours. The precipitate was removed by centrifugation to obtain a crude Lycium barbarum polysaccharide extract. The extract was purified by DEAE-52 cellulose column chromatography, and the polysaccharide fraction was eluted and collected. After vacuum concentration and spray drying, a light yellow powdered Lycium barbarum polysaccharide extract was obtained.

[0039] (3) Preparation of American ginseng extract: 500 g of American ginseng root segments were crushed and then added with 10 times the volume of purified water (i.e., 10 L). The extract was refluxed in a 95°C water bath for 3 times, each time for 1 hour. The combined filtrates were concentrated under reduced pressure. Ethanol was added to the concentrate to 70% (v / v) and allowed to stand for 12 hours to precipitate impurities. The supernatant of the alcohol precipitation was further enriched with saponin components by macroporous resin chromatography. The eluate was concentrated under reduced pressure and spray-dried to obtain a light yellow powdered American ginseng extract containing monomeric saponins such as Rg1 and Rb1.

[0040] (4) Take 100 g of the olive extract obtained above, 100 g of wolfberry extract, and 20 g of American ginseng extract, and mix them with appropriate amounts of skim milk powder (120 g), microcrystalline cellulose (90 g), mannitol (50 g), citric acid (5 g), steviol glycoside (2 g), aspartame (1.5 g), mint powder (1.0 g), and magnesium stearate (1.0 g) in proportion.

[0041] The above raw materials were added to a multi-dimensional motion mixing device, and the mixing time was set to 30 minutes and the stirring speed was set to 50 rpm to fully mix the ingredients to obtain a uniform compound powder.

[0042] The mixed powder was fed into a tableting machine for dry compression and compaction into chewable tablets with a mass of 0.7 g and a thickness of 3 mm. Each tablet contained 110 mg of olive extract, 110 mg of wolfberry extract, and 20 mg of American ginseng extract.

[0043] The resulting tablets are light brown, round tablets with a smooth, well-compacted surface, a refreshing taste, and a short disintegration time (2.5 minutes). The product is packaged in a sealed aluminum foil bag and stored in a dry, dark environment. It is suitable for daily use by people experiencing high altitude hypoxia, chronic fatigue, or sub-health conditions.

[0044] Example 2: Normal pressure closed hypoxia survival test 1 Materials and Methods 1.1 Experimental drugs Compound olive chewable tablets YJJ were provided by the Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences (prepared according to the method in Example 1) and stored in the Key Laboratory of Chemistry of Northwest Specialty Plant Resources, Chinese Academy of Sciences, and stored at 4°C. Hongyi capsules were purchased from Hubei Kangqin Pharmaceutical Co., Ltd. and stored in the Key Laboratory of Chemistry of Northwest Specialty Plant Resources, Chinese Academy of Sciences, and stored at 4°C.

[0045] 1.2 Experimental Animals One hundred SPF-grade Balb / C male mice (weighing 20–25 g) were purchased from the Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The animals had free access to food and water during the experiment and were housed in a 12-h light / 12-h dark environment with a room temperature of 23–28°C and a relative humidity of 40%–60%. All animal experiments were conducted in accordance with animal ethics guidelines and approved by the ethics committee.

[0046] 1.3 Experimental Methods 1.3.1 Animal grouping and drug administration One hundred mice were randomly divided into five groups (n = 20 / group), including a blank control group (NC), a positive control group (YX, Hongyi capsule), a high-dose group (SH), a medium-dose group (SM), and a low-dose group (SL) of compound olive chewable tablets. The mice were numbered and grouped by ear-tagging their left ears. As shown in Table 1, all test samples were measured at 0.01 mL∙g ~1 The volume was administered orally for 15 and 30 days respectively.

[0047] 1.3.2 Effect of Compound Olive Chewable Tablets on the Survival Time of Mice Under Normobaric Hypoxia Each dose group was orally administered with varying concentrations of the test sample, while the control group received the same volume of solvent. 15 days and 30 days after the first dose, and 1 hour after the last oral administration, mice in each group were placed in 250 mL ground-mouth bottles containing 5 g of soda lime (as a carbon dioxide absorber). The bottles were sealed with vaseline and tightly capped to prevent air leakage. Time was immediately measured, and the onset of systemic convulsions, muscle relaxation, and complete cessation of respiratory movement were used as the criteria for cessation of life. The time of death due to hypoxia was also recorded.

[0048] 1.3.3 Effect of Compound Olive Chewable Tablets on the Survival Rate of Mice Under Normobaric Hypoxia The survival time extension rate of each group under hypoxic conditions was calculated using the following formula: Among them, A0 is the average survival time of mice in the blank control group; A1 is the average survival time of mice in the drug-treated group.

[0049] 1.3.4 Determination of oxidative stress indicators in mouse blood The extracted blood supernatant was used to determine the levels of LDH, SOD, MDA, ATP and NO in serum according to the kit instructions.

[0050] 1.3.5 Determination of oxidative stress indicators in mouse heart and brain tissues Brain and heart tissues from each group of mice were rinsed with pre-chilled PBS, weighed, and homogenized in PBS at a ratio of 1:9. The supernatant was collected and centrifuged at 4°C (3000 rpm for 10 min). LDH, MDA, SOD, ATP, and NO were measured according to the kit instructions. The entire procedure was performed at low temperatures, and the samples were stored at −80°C until use.

[0051] 1.3.6 Data Processing All experiments were performed in triplicate, and the results are expressed as mean ± standard deviation. Origin 2021 software was used for data significance analysis and graphing.

[0052] 2. Experimental Results 2.1 Effect of Compound Olive Chewable Tablets on Survival Time of Mice The normobaric hypoxia model is an important means to evaluate the hypoxia tolerance of drugs. The survival time of mice in a closed hypoxic environment can directly reflect their tolerance to hypoxia. Figure 1 The experimental results showed that compared with the blank control group (NC), the positive control group (YX) showed a significant extension of survival time at both 15 days and 30 days, verifying the effectiveness of the model construction.

[0053] After intervention with compound olive chewable tablets, all dose groups demonstrated varying degrees of improvement, showing dose- and time-dependent trends. At 15 days, survival was significantly prolonged in the medium- and high-dose groups (SM and SH), with the SH group showing a more pronounced effect and the SL group showing limited improvement. After 30 days of intervention, mean survival times in the SH and SM groups reached 34.62 and 35.58 minutes, respectively, significantly higher than those in the NC group and comparable to or even slightly higher than those in the YX group (35.44 minutes). The SL group also showed a relatively stable prolongation trend. Significance analysis revealed statistically significant differences between the SH and SM groups and the NC group at both time points, further demonstrating that this intervention effectively improved mice's tolerance to hypoxia.

[0054] 2.2 Effect of Compound Olive Chewable Tablets on the Survival Rate of Mice To further quantify the intervention effect of the compound olive chewable tablets, this study calculated the survival extension rate for each group relative to the blank control group during the same period. The results, shown in Table 2, show that after 30 days of intervention, the survival extension rates in the SH and SM groups were 23.1% and 26.4%, respectively, exceeding those in the positive control group (YX, 25.9%). The SL group also achieved a survival extension rate of 16.4%. At 15 days, the survival extension rates were generally lower, but continued to show a dose-increasing trend, reaching 18.9% in the SH group, 16.4% in the SM group, and 11.6% in the SL group.

[0055] This result is consistent with the statistical trend of survival time, suggesting that the compound olive chewable tablets enhance hypoxic survival in a time-accumulating and dose-dependent manner. The hypoxia tolerance of the medium- and high-dose groups after 30 days of continuous intervention approached or even exceeded that of the positive control group, demonstrating promising pharmacological activity and potential for application.

[0056] 2.3 Oxidative stress indicators in mouse blood Under hypoxic conditions, serum oxidative stress levels can directly reflect the body's systemic stress response. Key indicators such as lactate dehydrogenase (LDH), superoxide dismutase (SOD), malondialdehyde (MDA), adenosine triphosphate (ATP), and nitric oxide (NO) can be measured to assess the antioxidant and tissue-protective effects of compound olive chewable tablets.

[0057] 2.3.1 Effect of Compound Olive Chewable Tablets on LDH Activity in Mice Blood LDH is a sensitive indicator of cell damage, and its increase usually indicates increased cell membrane permeability or necrosis tendency. Figure 2 The SH group significantly reduced serum LDH levels at both 15 and 30 days. The 30-day level was significantly lower than that of the NC group, and showed no significant difference from the YX group, or even slightly better. The SM group also exhibited a certain degree of LDH inhibition. This suggests that the compound olive chewable tablet can effectively alleviate hypoxia-induced cell damage and possesses promising tissue protective potential.

[0058] 2.3.2 Effect of Compound Olive Chewable Tablets on SOD Activity in Mice Blood SOD is a key enzyme in the endogenous antioxidant defense system that can efficiently remove O2 - Free radicals. Data such as Figure 3 Results showed that both the SH and SM groups significantly increased serum SOD activity at both 15 and 30 days. The SH group had the highest level at 30 days, similar to that of the YX group and statistically significantly different from the NC group. However, the SL group did not show a significant increase. These results suggest that the compound olive chewable tablets can enhance the overall antioxidant capacity of mice and strengthen the physiological defense mechanism against oxidative stress by increasing SOD activity.

[0059] 2.3.3 Effect of Compound Olive Chewable Tablets on MDA Levels in Mice Blood MDA is the end product of lipid peroxidation and reflects the degree of oxidative damage to the cell membrane structure. Figure 4 As shown, the MDA content in the SH group was significantly lower than that in the NC group at 30 days and comparable to that in the YX group. The SM group also showed a downward trend. This suggests that long-term intervention with high-dose compound olive chewable tablets can effectively inhibit lipid oxidation and maintain cell membrane stability.

[0060] 2.3.4 Effect of Compound Olive Chewable Tablets on ATP Content in Mice Blood ATP is a key indicator of cellular energy metabolism, and hypoxia often leads to ATP depletion. Figure 5 As shown, both the SH and SM groups showed a significant increase in ATP content after 15 and 30 days. The SH group, in particular, showed a significantly higher ATP content than the NC group at 30 days (p < 0.001), and slightly better than the YX group. This result suggests that the compound olive chewable tablet can improve energy metabolism disorders, enhance mitochondrial function, and delay energy depletion.

[0061] 2.3.5 Effect of Compound Olive Chewable Tablets on NO Level in Mice Blood NO is a product of the nitric oxide synthase system and has dual functions of vasodilation and anti-oxidation. Figure 6 Results showed that the SH group significantly increased serum NO levels at both 15 and 30 days, with the greatest increase on day 30, slightly better than the YX group and significantly different from the NC group. The SM group also showed some improvement. This suggests that the compound olive chewable tablets may improve microcirculation by increasing NO levels, helping to alleviate ischemic hypoxia.

[0062] Compound olive chewable tablets demonstrated excellent anti-hypoxic activity in a normobaric hypoxia animal model, significantly prolonging mouse survival with a clear dose-dependent and time-dependent enhancement effect. After 30 days of intervention, the survival rates of the medium- and high-dose groups were 26.4% and 23.1%, respectively, exceeding those of the blank group and comparable to those of the positive control group. The chewable tablets also effectively modulated serum levels of LDH, MDA, SOD, NO, and ATP, demonstrating their multifaceted effects in scavenging free radicals, stabilizing energy metabolism, and protecting tissue function.

[0063] The comprehensive results show that the preparation is of natural origin, has good safety, and has physiological effects against hypoxia. It is suitable for development as a hypoxia-resistant functional food or an auxiliary intervention product for chronic hypoxia.

[0064] Example 3: Comparison of single-component chewable tablets and compound chewable tablets In order to verify the synergistic anti-hypoxia synergistic effect among the olive extract, wolfberry extract and American ginseng extract in the present invention, the following control group was designed and prepared and animal experiments were conducted.

[0065] 1 Materials and Methods 1.1 Experimental drugs Preparation of single olive extract chewable tablets: 100 g of the olive extract obtained above was taken and mixed with appropriate amounts of skim milk powder (120 g), microcrystalline cellulose (90 g), mannitol (50 g), citric acid (5 g), steviol glycoside (2 g), aspartame (1.5 g), mint powder (1.0 g) and magnesium stearate (1.0 g) in proportion.

[0066] The above raw materials were added to a multi-dimensional motion mixing device, and the mixing time was set to 30 minutes and the stirring speed was set to 50 rpm to fully mix the ingredients to obtain a uniform compound powder.

[0067] The mixed powder was fed into a tableting device for dry pressing and compacting into chewable tablets with a single tablet weight of 0.7 g and a thickness of 3 mm.

[0068] Preparation of single wolfberry extract chewable tablets: 100 g of the wolfberry extract obtained above was taken and mixed with appropriate amounts of skim milk powder (120 g), microcrystalline cellulose (90 g), mannitol (50 g), citric acid (5 g), steviol glycoside (2 g), aspartame (1.5 g), mint powder (1.0 g) and magnesium stearate (1.0 g) in proportion.

[0069] The above raw materials were added to a multi-dimensional motion mixing device, and the mixing time was set to 30 minutes and the stirring speed was set to 50 rpm to fully mix the ingredients to obtain a uniform compound powder.

[0070] The mixed powder was fed into a tableting device for dry pressing and compacting into chewable tablets with a single tablet weight of 0.7 g and a thickness of 3 mm.

[0071] Preparation of single American ginseng extract chewable tablets: 100 g of the American ginseng extract obtained above was taken and mixed with appropriate amounts of skim milk powder (120 g), microcrystalline cellulose (90 g), mannitol (50 g), citric acid (5 g), steviol glycoside (2 g), aspartame (1.5 g), mint powder (1.0 g) and magnesium stearate (1.0 g) in proportion.

[0072] The above raw materials were added to a multi-dimensional motion mixing device, and the mixing time was set to 30 minutes and the stirring speed was set to 50 rpm to fully mix the ingredients to obtain a uniform compound powder.

[0073] The mixed powder was fed into a tableting device for dry pressing and compacting into chewable tablets with a single tablet weight of 0.7 g and a thickness of 3 mm.

[0074] 1.2 Experimental Animals Eighty SPF-grade Balb / C male mice (weighing 20-25 g) were purchased from the Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The animals had free access to food and water during the experiment and were housed in a 12-h light / 12-h dark environment with a room temperature of 23-28°C and a relative humidity of 40%-60%. All animal experiments were conducted in accordance with animal ethics guidelines and approved by the ethics committee.

[0075] 1.3 Experimental Methods 1.3.1 Animal grouping and drug administration Eighty mice were randomly divided into four groups (n=20 / group), including single olive extract chewable tablets, single wolfberry extract chewable tablets, single American ginseng extract chewable tablets, and compound olive extract chewable tablets. They were numbered and grouped by ear tagging their left ears. All test samples were measured at 0.01 mL∙g ~1 The dose of 192.0 mg / kg / d was administered orally for 30 days.

[0076] 1.3.2 Effect of chewable tablets on the survival time of mice under normobaric hypoxia Each dose group was orally administered with varying concentrations of the test sample, while the control group received the same volume of solvent. Thirty days after the first dose and one hour after the final oral administration, mice in each group were placed in a 250 mL ground-mouth bottle containing 5 g of soda lime (as a carbon dioxide absorber) (one mouse per bottle). The bottle was sealed with vaseline and tightly capped to prevent air leakage. Time was immediately measured, and the onset of systemic convulsions, muscle relaxation, and complete cessation of respiratory movement in the mouse was used as the criterion for death. The time of death due to hypoxia was also recorded.

[0077] 1.3.3 Effect of chewable tablets on the survival rate of mice subjected to normobaric hypoxia The survival time extension rate of each group under hypoxic conditions was calculated using the following formula: Among them, A0 is the average survival time of mice in the blank control group; A1 is the average survival time of mice in the drug-treated group.

[0078] 1.3.4 Determination of oxidative stress indicators in mouse blood Using the extracted blood supernatant, the serum levels of LDH, SOD, MDA, ATP, and NO were determined according to the kit instructions. After each group of indicators was determined, the change rate (%) of each treatment group relative to the blank group was calculated using the following formula: Where A0 is the average value of the corresponding indicator in the blank control group; A1 is the average value of the corresponding indicator in the olive group, wolfberry group, American ginseng group, or compound group. The calculated results were rounded to two decimal places for subsequent statistical analysis and evaluation of synergistic effects.

[0079] 1.3.3 Data Processing All experiments were performed in triplicate, and the results are expressed as mean ± standard deviation. Origin 2021 software was used for data significance analysis and graphing.

[0080] 3. Experimental Results The results showed that all three single components had a certain degree of prolonging effect on the hypoxia survival of mice, among which the olive group performed best, with a survival extension rate of 16.41%, while the wolfberry group and American ginseng group were 11.54% and 13.67%, respectively. In comparison, the survival extension rate of the compound group of the present invention reached 26.48%, which was significantly higher than that of each single component. The experimental results further proved that the combination of olive extract, wolfberry extract and American ginseng extract has a synergistic effect. The hypoxia resistance effect of the compound group is not only better than that of any single component, but also the increase is significantly greater than the improvement of the single component group, reflecting the multi-target and multi-mechanism compound effect.

[0081] These results showed that the olive group significantly improved SOD activity (+7.33%) and also reduced MDA levels (-9.17%), suggesting a potential antioxidant effect. The wolfberry group showed the most significant improvement in NO levels (-4.50%) and also reduced LDH levels (-9.71%), demonstrating its effects on vasodilation and cell damage protection. The American ginseng group had the most significant effects on LDH (-11.69%) and ATP (+1.56%), suggesting that its effects primarily occur through improving cellular energy metabolism and inhibiting cell damage. However, while the three individual components demonstrated some improvement in various indicators, none reached the level of comprehensive improvement achieved by the compound formula of the present invention. The compound formula significantly outperformed either single component in terms of increasing SOD activity (+15.64%), reducing MDA levels (-14.99%), reducing LDH levels (-18.39%), increasing NO levels (+13.14%), and increasing ATP content (+4.98%). These results fully demonstrate that the compound formula has significant advantages in antioxidant capacity, cell integrity protection, vascular regulation, and energy metabolism support, reflecting a systemic synergistic effect of "multi-target, multi-mechanism."

Claims

1. A compound chewable tablet with anti-hypoxia activity, characterized in that: The composition comprises the following components in weight percentage: Olive extract: 15%-25%; Lycium barbarum extract: 15%-25%; American ginseng extract: 2%-6%; Skimmed milk powder: 18%-28%; Microcrystalline cellulose: 15%-22%; Mannitol: 8%-12%; Citric acid: 0.5% to 1.5%; Sweetener: 0.2%~0.8%; Mint powder: 0.1%~0.5%; Tableting excipients ≤0.5%; The olive extract, wolfberry extract and American ginseng extract enhance the anti-hypoxia activity through synergistic action.

2. The compound chewable tablet according to claim 1, characterized in that The olive extract contains hydroxytyrosol and oleuropein; the wolfberry extract contains wolfberry polysaccharide; and the American ginseng extract contains ginsenosides Rb1 and Rg1.

3. The compound chewable tablet according to claim 1, characterized in that The sweetener is one or both of stevioside and aspartame; the tableting auxiliary material is one or both of silicon dioxide and magnesium stearate.

4. The compound chewable tablet according to claim 1, characterized in that The mass of a single chewable tablet is 0.5 to 1.2 g, and each tablet contains no less than 100 mg of the active ingredient of the olive extract.

5. The compound chewable tablet according to claim 1, characterized in that The chewable tablets are prepared by direct compression and have a disintegration time of ≤3 minutes; The chewable tablet further comprises one or more of natural flavors, plant pigments, oral lubricants or food-grade adhesives to improve taste and appearance.

6. The method for preparing the compound chewable tablet according to any one of claims 1 to 5, characterized in that: The following steps are involved: (1) Mix the components in proportion and stir at 45-65 rpm for 20-40 minutes; (2) The mixed powder is dry-pressed into tablets weighing 0.5 to 1.2 g; (3) Store the tablets sealed and away from light.

7. Use of the compound chewable tablet according to any one of claims 1 to 5 in the preparation of anti-hypoxia functional food or health care product, characterized in that: The compound chewable tablet exerts an anti-hypoxia function by increasing the levels of superoxide dismutase, nitric oxide and adenosine triphosphate in the body; The compound chewable tablet exerts an anti-hypoxia function by reducing lactate dehydrogenase and malondialdehyde oxidative stress indicators.

8. The use according to claim 7, characterized in that: The anti-hypoxia functional food or health product is suitable for improving and intervening in plateau hypoxia, mental fatigue, chronic fatigue syndrome or senile hypoxia.

9. The use according to claim 7 or 8, characterized in that: The compound chewable tablet achieves anti-hypoxia effect by regulating oxidative stress pathways, improving mitochondrial metabolism and inhibiting cell apoptosis.

10. An anti-hypoxia functional food or health product, characterized by: The compound chewable tablet comprises the compound chewable tablet according to any one of claims 1 to 5, with a daily intake of 1 to 4 tablets.

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