Use of tyrosol sulfate sodium and derivative tyrosol compounds in the preparation of brain health products

Abstract

The application belongs to the technical field of biological medicine, and provides application of tyrosol sodium sulfate and derivative tyrosol compounds in preparation of brain health products. The structure of the tyrosol sodium sulfate and the derivative tyrosol compounds is shown as formula I, which can effectively reduce the infarction area after cerebral ischemia, and reduce cerebral edema caused by cerebral ischemia, and has wide application in resisting cerebral ischemia and improving cerebral edema and cerebral infarction caused by cerebral ischemia. Meanwhile, the tyrosol sodium sulfate and the derivative tyrosol compounds can effectively improve learning and memory ability, promote recovery of brain function, and have wide application in treating behavior / cognitive dysfunction diseases such as brain trauma, cerebrovascular accident, Alzheimer's disease and Parkinson's disease. Formula I.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to application of tyrosol sodium sulfate and a derivative tyrosol compound in preparation of brain health products. BACKGROUND

[0002] Cognitive dysfunction is one of the common complications of various chronic diseases, such as stroke, cerebral palsy, diabetes, etc. The incidence is high, and early identification and intervention are crucial for rehabilitation. Clinical treatment includes: increasing cerebral blood flow, improving cerebral ischemia; improving brain oxygen supply; inhibiting lipid peroxidation, removing free radicals, etc. Commonly used drugs include dihydroergotoxine preparations, neurotrophic drugs, calcium ion antagonists, etc., but there is still a lack of evidence-based medical evidence.

[0003] Tyrosol (p-hydroxyphenyl ethanol) is first isolated from olive oil, and is one of the important antioxidant components in olive oil with relatively high content. In addition, it is also widely present in plants such as rhodiola, ligustrum, grape seeds, etc., and is the aglycone component of the main active ingredient of rhodiola in traditional Chinese medicine, rhodiola glycoside. Its chemical structure has substitution reaction on the benzene ring and some reaction performance of hydroxyl group, and a series of derivative tyrosol compounds can be prepared by biosynthesis and chemical synthesis. Among them, hydroxytyrosol is the most widely studied.

[0004] For example, Chinese patent application CN101674817A discloses that hydroxytyrosol can enhance chondrocyte proliferation, improve extracellular matrix synthesis, and is used as an effective micronutrient for cartilage repair and regeneration. Chinese patent CN103961338B discloses the application of hydroxytyrosol in anti-doxorubicin cardiotoxicity, and hydroxytyrosol has significant resistance to cardiotoxicity induced by doxorubicin and has strong protective effect on myocardial cells.

[0005] For example, Chinese patent application CN105213356A discloses the application of hydroxytyrosol in the preparation of anti-vascular dementia drugs, which verifies that hydroxytyrosol can reduce neurological deficits, reduce the range of cerebral infarction and reduce the degree of cerebral edema, effectively inhibit the morphological changes of nerve tissue in vascular dementia rats, improve the cognitive, learning and memory abilities of vascular dementia rats, and reduce the symptoms of dementia.

[0006] However, tyrosol or hydroxytyrosol has limited effect on the improvement of cognitive dysfunction caused by cerebral ischemia. The research on tyrosol derivatives without hydroxyl substitution at position 3 of the benzene ring is relatively less. Studies have shown that tyrosol and its derivatives such as tyrosol sodium sulfate, tyrosol glucuronide, and tyrosol fatty acid ester also have anti-inflammatory, antioxidant stress, etc. However, its use still needs to be further developed. SUMMARY

[0007] In order to solve the above problems, the application provides application of tyrosol sodium sulfate and derivative tyrosol compounds in preparation of brain health products. The application of tyrosol sodium sulfate and derivative tyrosol compounds in anti-cerebral ischemia, anti-cerebral anoxia, improvement of learning and memory ability, improvement of cerebral edema caused by cerebral ischemia and other improvement of brain function health food or pharmaceutical preparations is first proposed.

[0008] In order to achieve the above-mentioned purpose, the technical scheme of the application is as follows:

[0009] The application provides application of tyrosol sodium sulfate and derivative tyrosol compounds in preparation of brain health products.

[0010] Further, the tyrosol sodium sulfate and derivative tyrosol compounds have the following structural general formula:

[0011]

[0012] In the formula, R1 (phenolic hydroxyl substituent) is C1-C18 alkyl, -COR', -SO3Na, sugar radical or glucuronic acid; R2 (alcoholic hydroxyl substituent) is H or -COR''; R' and R'' are each independently selected from any one of C1-C18 alkyl (i.e. -COR' is specifically: formic acid, acetic acid, fumaric acid, succinic acid, tartaric acid, octanoic acid, decanoic acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid and other tyrosol ester compounds formed with phenolic hydroxyl; -COR'' is specifically: formic acid, acetic acid, fumaric acid, succinic acid, tartaric acid, octanoic acid, decanoic acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid and other tyrosol ester compounds formed with alcoholic hydroxyl).

[0013] Further, the sugar radical is a radical after removal of hydroxyl groups of glucose, galactose, arabinose or rhamnose (i.e. -OR1 is specifically a glycoside formed by glucose, galactose, arabinose or rhamnose and phenolic hydroxyl of tyrosol).

[0014] In some embodiments of the application, the tyrosol sodium sulfate and derivative tyrosol compounds are compounds in which only the alcoholic hydroxyl position is substituted, i.e. in the general formula, R1 is any one of C1-C18 alkyl, -COR', -SO3Na, sugar radical or glucuronic acid; R2 is H, wherein R' is C1-C18 alkyl.

[0015] Preferably, it is: , , , or .

[0016] In some embodiments of the present application, the tyrosol sodium sulfate and the derivative tyrosol compound is a compound in which both the alcohol hydroxyl and the phenolic hydroxyl are substituted, i.e., in the general formula, R1 is any one of C1-C18 alkyl, -COR', -SO3Na, a sugar group, or glucuronic acid; and R2 is -COR''.

[0017] Preferably, R1 is -SO3Na; R2 is -COR''; and R'' is any one of C1-C18 alkyl (i.e., -COR' is specifically formic acid, acetic acid, fumaric acid, succinic acid, tartaric acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, etc.).

[0018] Further preferably, .

[0019] Further, the brain health product is any one of an anti-cerebral ischemia product, an anti-cerebral edema product caused by cerebral ischemia, an anti-cerebral infarction product caused by cerebral ischemia, an anti-cerebral anoxia product, and an anti-behavior / cognitive dysfunction product.

[0020] Still further, the behavior / cognitive dysfunction includes any one of brain trauma, cerebral vascular accident, Alzheimer's disease, and Parkinson's disease.

[0021] Further, the brain health product is a pharmaceutical preparation or a health food.

[0022] The present application also provides a pharmaceutical preparation, wherein the active ingredient of the pharmaceutical preparation comprises a tyrosol sodium sulfate and a derivative tyrosol compound; and the tyrosol sodium sulfate and the derivative tyrosol compound have the following general formula:

[0023]

[0024] wherein R1 is C1-C18 alkyl, -COR', -SO3Na, a sugar group, or glucuronic acid; R2 is H or -COR''; R' and R'' are each independently any one of C1-C18 alkyl; and the sugar group is a group obtained by removing a hydroxyl group from glucose, galactose, arabinose, or rhamnose.

[0025] Further, the pharmaceutical preparation further comprises one or more of a pharmaceutically acceptable salt, a solvate, a hydrate, and a polymorph of the derivative tyrosol compound.

[0026] Further, the pharmaceutical preparation further comprises a pharmaceutically acceptable excipient.

[0027] Further, the pharmaceutical preparation is any one of a tablet, a capsule, a pill, a granule, a powder, a paste, a mixture, a suspension, an injection, and a transdermal absorption agent.

[0028] The present application also provides a brain health care food, which comprises tyrosol sodium sulfate and derivative tyrosol compounds; the tyrosol sodium sulfate and derivative tyrosol compounds have the following general structure:

[0029]

[0030] wherein R1 is C1-C18 alkyl, -COR', -SO3Na, a sugar group or a glucuronic acid; R2 is H or -COR''; R' and R'' are each independently selected from any one of C1-C18 alkyl; the sugar group is a group of glucose, galactose, arabinose or rhamnose after losing a hydroxyl group.

[0031] Further, the brain health care food further comprises a food acceptable auxiliary material.

[0032] Compared with the prior art, the present application has the following beneficial effects:

[0033] The tyrosol sodium sulfate and derivative tyrosol compounds (especially the derivative tyrosol compounds C and E containing sulfate groups) provided by the present application have more remarkable effects in anti-cerebral ischemia, relieving cerebral infarction, reducing cerebral edema, anti-cerebral anoxia and improving learning and memory compared with tyrosol, and have wide application in products such as anti-cerebral ischemia, anti-cerebral anoxia, improving cerebral edema and cerebral infarction caused by cerebral ischemia, and improving behavioral / cognitive dysfunction. BRIEF DESCRIPTION OF DRAWINGS

[0034] Figure 1 Results of the influence of compound C on the neurological behavior score of MCAO rats;

[0035] Figure 2 Results of the influence of compound C on cerebral infarction of MCAO rats;

[0036] Figure 3 Results of the influence of compound C on cerebral edema of MCAO rats;

[0037] Figure 4 Results of the influence of compound C on the escape latency of MCAO rats in the Morris water maze experiment;

[0038] Figure 5 Results of the influence of compound C on the platform crossing times of MCAO rats in the Morris water maze experiment;

[0039] Figure 6 Results of the influence of compound C on the time ratio of MCAO rats in the target quadrant in the spatial exploration experiment in the Morris water maze experiment;

[0040] Figure 7Figure 5 is a swimming route diagram of the escape latency of the rats in the Morris water maze experiment on the 5th day of the sham group, the MCAO group and the administration group (compound C);

[0041] Figure 8 Results of the protective effect of tyrosol sodium sulfate and the derived tyrosol compounds on neurons; wherein, compared with the normal group, * p <0.05, ** p <0.01; compared with the model group, # p <0.05, ## p <0.01;

[0042] Figure 9 Results of the protective effect of tyrosol sodium sulfate and the derived tyrosol compounds on astrocytes; wherein, compared with the normal group, * p <0.05, ** p <0.01; compared with the model group, # p <0.05, ## p <0.01. DETAILED DESCRIPTION

[0043] In order to make the purposes, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor fall within the protection scope of the present application.

[0044] The present application verifies the effect of improving brain health of tyrosol sodium sulfate and the derived tyrosol compounds by taking compounds A-H in Table 1 as experimental drugs.

[0045] Table 1 Structure information of tyrosol sodium sulfate and the derived tyrosol compounds

[0046]

[0047] Compound E was prepared according to the method reported in the literature Exploring the Biological Potential of Hydroxytyrosol and Derivatives: Synthetic Strategies and Evaluation of Antiproliferative, Antioxidant, and Antimicrobial Activities (J. Agric. Food Chem. 2024, 72, 48, 26699-26710).

[0048] Experimental Example 1: Study on the protective effect of tyrosol sodium sulfate and its derivative tyrosol compounds on the brain nerves of rats in middle cerebral artery occlusion (MCAO) model rats

[0049] (1) Experimental animals: male SD rats, SPF level, body weight 220±10 g.

[0050] (2) Experimental method: 4% chloral hydrate was injected intraperitoneally for anesthesia, and a plug line was inserted into the internal carotid artery. After 90 min of plug insertion, reperfusion was performed, and blood could re-enter the middle cerebral artery through the willis circle to achieve cerebral vascular reperfusion.

[0051] The experimental rats were scored twice according to the principle of random blind method. The rats were scored for the first time 2 h after surgery, and scored for the second time 24 h later. Referring to the 5-point scoring standard of Longa et al., the specific scoring criteria are as follows:

[0052] 0 points: normal, no neurological signs;

[0053] 1 point: animal cannot fully extend left forelimb;

[0054] 2 points: animal left limb paralysis, walking to the left side, and appearing tail chasing phenomenon;

[0055] 3 points: animal walking to the left side falls, or animal cannot stand or roll;

[0056] 4 points: no spontaneous activity, and conscious disturbance.

[0057] The neurological deficit score of 1-3 points can be evaluated as successful modeling. Except for the sham operation group, 0 points (model unsuccessful) and 4 points (damage too severe, died within 24 h) in each group will be excluded, and animals in the excluded group will be supplemented in the subsequent test to ensure the number of animals in each group.

[0058] (3) Experimental grouping and administration: the successfully modeled rats were randomly divided into 10 groups, and administration was performed immediately after the insertion of the online plug. Low-dose (10 mg / kg) and high-dose (20 mg / kg) of compounds A-H (prepared in normal saline with a concentration of 10 mg / ml) were administered by intraperitoneal injection, and the sham operation group (Sham) and the model group (MCAO) were administered with the same amount of normal saline. The rats were sacrificed 24 hours later.

[0059] (4) Experimental results:

[0060] 4.1 The results of the neurological behavior score of the MCAO rats are shown in Table 2. The neurological behavior scores of the sham operation group, the model group, and the low-dose and high-dose compound C groups after 2 hours and 24 hours are compared as shown in Figure 1 .

[0061] Table 2. Results of the neurological behavior score of rats in each group (n=10)

[0062]

[0063] Note: Compared with the sham operation group, ** p <0.01; compared with the model group, # p <0.05, ## p <0.01.

[0064] As can be seen from the above table: compared with the MCAO group, compounds B-H can reduce the neurological behavior score of the cerebral ischemic rats, and among them, the effect of compound C and compound E in reducing the neurological behavior score of the cerebral ischemic rats after 2 hours of administration at a dose of 10 mg / kg is significantly better than that of compound A p (<0.01).

[0065] 4.2 Cerebral infarction area: 24 hours after the MCAO operation of the rats, the rats were anesthetized by intraperitoneal injection of chloral hydrate, quickly decapitated and sacrificed, the skull was opened to take the brain, the surface blood and cerebrospinal fluid was gently absorbed with filter paper, and the brain slices were immediately dyed with 1% TTC. Photographs of the distribution of the infarction area of the brain slices were taken, wherein the TTC staining results of the sham operation group, the model group, and the low-dose and high-dose compound C groups are shown in Figure 2 A of FIG. 1. The ischemic area and the total brain slice area of each brain slice were drawn using image proplus software, and the cerebral infarction area was calculated. The cerebral infarction area (%) = cerebral infarction area / total brain slice area x 100%, and the results are shown in Table 3. The TCC staining diagrams of the sham operation group, the MCAO model group, and the low-dose and high-dose compound C groups are shown in Figure 2 B of FIG. 1.

[0066] Table 3 Results of cerebral infarction area in each group of rats ( (n=10)

[0067]

[0068] Note: Compared with the sham surgery group, ** p <0.01; compared with the model group, # p <0.05, ## p <0.01.

[0069] Depend on Figure 2 As shown in the table above, significant cerebral infarction foci are observed after cerebral ischemia. Compared with the MCAO group, compound BH can reduce the infarct area. Among them, compound CH can significantly reduce the cerebral infarction area at a dosage of 10 mg / kg. p <0.05 or p <0.01), indicating that compounds CH have a better effect on improving cerebral infarction, especially compound E.

[0070] Experimental Example 2: Study on the improvement of cerebral edema in rats with middle cerebral artery occlusion (MCAO) model by sodium tyrosol sulfate and its derivative tyrosol compounds.

[0071] The experimental animals, experimental methods, and grouping of drugs were the same as in Experiment 1.

[0072] Rats were sacrificed 24 hours after model establishment and drug administration. The whole brain was harvested and weighed wet. After baking at 60℃ for 24 hours, the dry weight was measured. The water content of the brain tissue was calculated using the wet-dry method. The rat body weight and whole brain wet weight were obtained using an electronic balance (accurate to 0.001g). Brain index (%) = whole brain wet weight / rat body weight × 100%; brain water content (%) = (wet weight - dry weight) / wet weight × 100%. The results are shown in Table 4. The comparison of brain edema between the sham-operated group, the model group, and the low-dose and high-dose groups of compound C is as follows: Figure 3 As shown.

[0073] Table 4. Results of brain index and brain water content in each group of rats ( (n=10)

[0074]

[0075] Note: Compared with the sham surgery group, ** p <0.001; compared with the model group, # p <0.05, ## p <0.01.

[0076] From the above table, it can be seen that compared with the MCAO group, compounds C-H showed different degrees of effect in reducing brain edema caused by local cerebral ischemia, while reducing the brain index; among them, compounds C, D, E and H are better (10 mg / kg administration dose, p <0.01), compounds F and G are less.

[0077] Experimental Example 3: Effect of tyrosol sodium sulfate and its derivative tyrosol compounds on learning and memory of rats with multiple cerebral infarction

[0078] (1) Experimental animals: male SD rats, SPF level, body weight 220±10g.

[0079] (2) Experimental method: Morris water maze experiment, 4% chloral hydrate was injected intraperitoneally for anesthesia, 0.2 mL of fluorescent microspheres was injected from the external carotid artery to the internal carotid artery with a syringe, and then the artery clamp was loosened. At this time, the fluorescent microspheres will disperse from the internal carotid artery with the blood flow to each artery of the brain. After the operation, low-dose (10 mg / kg) and high-dose (20 mg / kg) compounds A-H (prepared with normal saline to a concentration of 10 mg / ml) were given by intraperitoneal injection for 28 consecutive days. After 28 days, the Morris water maze experiment was performed for the acquired training. In the training test, each rat needed to receive training in the first, second, third and fourth quadrants every day, and the order was random every day.

[0080] At the beginning of each quadrant test, the rat was placed in the water with its face to one side of the pool wall. Once the rat is in the water, it is allowed to locate the escape platform for 90 s; if it is not found within 120 s, the rat needs to be guided to the platform. Once the rat climbs onto the platform, it is allowed to rest for 10 s (if less than 10 s, it needs to be guided again) to observe the spatial cues on the platform. The time and distance traveled by the rat before finding the hidden platform are recorded using a video tracking system. After that, the rat is removed from the water maze, and after the training in the first quadrant is completed, the rat is released in a different quadrant, and each rat is guaranteed to have more than 5 min of rest time to recover body temperature and strength. For five consecutive days, the sixth day is the water maze exploration experiment. Take away the platform and put the rat in the third quadrant, and record the distance and time of the rat swimming through the first quadrant for 90 s. During this period, due to the rat's instinct to avoid water and seek survival, it will find the escape platform according to its memory in the acquisition experiment. The escape platform in the acquisition training is placed in the middle of the first quadrant.

[0081] (3) Experimental results:

[0082] The results of the Morris water maze experiment are shown in Tables 5-7. The escape latency, platform crossing times, time ratio in the target quadrant in the spatial probe test, and the swimming path of the escape latency of the rats in the Morris water maze experiment in the sham operation group, the model group, and the low-dose and high-dose compound groups are shown in Figures 4-7 .

[0083] Table 5. Escape latency of rats in the Morris water maze experiment (n = 10)

[0084]

[0085] Note: Compared with the sham operation group, ** p <0.01; compared with the model group, # p <0.05, ## p <0.01.

[0086] Table 6. Effect of zearalenol sodium sulfate and its derivative zearalenol compounds on the platform crossing times of cerebral ischemic rats (n = 10)

[0087]

[0088] Note: Compared with the sham operation group, ** p <0.01; compared with the model group, # p <0.05, ## p <0.01.

[0089] Table 7. Effect of zearalenol sodium sulfate and its derivative zearalenol compounds on the time ratio in the target quadrant of cerebral ischemic rats (n = 10)

[0090]

[0091] Note: Compared with the sham operation group, ** p <0.01; compared with the model group, # p <0.05, ## p <0.01.

[0092] From the above results, it can be seen that, as the test time increases, the escape latency of the MCAO group is significantly longer than that of the Sham group (starting from the first day, p<0.01). Compared with the MCAO group, in all treatment groups (except compound A), the escape latency of rats was significantly shortened with the increase of training sessions. At a dosage of 20 mg / kg, there were significant differences between the rats in each group and the model group from the second day onwards. p <0.05), among which compounds C and E were the best, followed by compounds B and D, and compounds F, G and H also showed varying degrees of shortening of escape latency in rats.

[0093] In addition, compared with the MCAO group, except for the low-dose groups of compound A and compound B, the number of rats in the other treatment groups was significantly reduced and the percentage of time spent in the platform quadrant was significantly increased. Moreover, the effects of each compound on reducing escape latency showed similar trends. Overall, compounds C, D, and E were the best, followed by F, G, and H.

[0094] Experimental Example 4: Protective Effects of Sodium Tyrosol Sulfate and its Derivative Tyrosol Compounds on Neurons

[0095] (1) Experimental method: Human neuroblastoma SH-SY5Y cells were used to conduct an oxygen-glucose deprivation (OGD) induction experiment.

[0096] OGD Induction Model: Sugar-free DMEM medium, pre-saturated with a 95% N2-5% CO2 mixture for 15 min, was added to each well of a 96-well plate. The plate was placed in a hypoxia chamber, and the 95% N2-5% CO2 mixture was circulated. The oxygen concentration at the outlet was monitored in real-time using an oxygen analyzer. When the oxygen concentration reached 1%, ventilation was stopped, the inlet and outlet pipes were closed, and the hypoxia chamber was placed in a cell culture incubator for 4 hours of hypoxic culture. Reoxygenation: The hypoxia chamber was opened, the 96-well plate was removed, the hypoxic solution was aspirated, and sugar-free AMEM medium was added to each well. The plate was then placed in an incubator for another 2 hours of culture. Sugar-free DMEM medium without hypoxia-reoxygenation treatment served as a blank control (normal group). Except for the model group, the treatment groups were given 25 μM, 50 μM, and 100 μM of compound AH, respectively, during hypoxia.

[0097] (2) Experimental results:

[0098] like Figure 8 As shown, after OGD induction, compared with the model group, compound E at a concentration of 25 μM significantly improved cell viability after injury. p <0.01%, compounds C, F, G, and H at 50 μM significantly improved cell viability after injury ( p <0.05); Compounds A, B, and D significantly ameliorated OGD-induced SH-SY5Y cell damage at a high concentration of 100 μM. p<0.05). The results showed that E had the strongest neuroprotective effect, followed by C, F, G and H, but all of them had lower effective concentrations than tyrosol and rhodioloside.

[0099] Example 5: Protective effect of sodium tyrosol sulfate and its derivative tyrosol compounds on astrocytes

[0100] (1) Experimental method: Human astrocytes SVGp12 were used for glucose-oxygen deprivation (OGD) induction experiment. The method was the same as in Experiment 4.

[0101] (2) Experimental results:

[0102] like Figure 9 As shown, after OGD induction, both compounds C and D at 50 μM significantly improved cell viability after injury. p <0.01%, compound E at 50 μM significantly improved cell viability after injury ( p <0.05); A, B, G, and F significantly protected astrocytes from hypoxic-hypoxic injury at 100 μM. p <0.05). The above results demonstrate that compounds C, D, and E of this invention have superior protective effects on astrocytes.

[0103] 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 sodium tyrosol sulfate and its derivative tyrosol compounds in the preparation of brain health products, characterized in that, The sodium tyrosol sulfate and the derived tyrosol compounds have the following general structural formula: Wherein, R1 is -SO3Na, R2 is H, and R2'' is -COR''; where R'' is a C1-C18 alkyl group; Alternatively, R1 may be methyl, -COR', glycosyl, or glucuronic acid, and R2 may be H; wherein R' is a C1-C18 alkyl group; and the glycosyl group may be glucose, galactose, arabinose, or rhamnose after the loss of a hydroxyl group. The brain health product is any one of the following: anti-cerebral ischemia, improvement of cerebral edema caused by cerebral ischemia, improvement of cerebral infarction caused by cerebral ischemia, and behavioral / cognitive dysfunction. The brain health product is a pharmaceutical preparation.

2. The application according to claim 1, characterized in that, The sodium tyrosol sulfate and the derived tyrosol compound are any of the following structures: , , , , or .

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