A composition, its preparation and use

Abstract

This invention discloses a pharmaceutical composition comprising a compound of formula I and dextroborneol, which has shown unexpected effects in treating cerebrovascular diseases and central nervous system diseases.

Description

Technical Field

[0001] This application relates to a pharmaceutical composition that can be developed for use as a medicine for treating diseases caused by oxidative stress and / or thrombosis induced by free radicals. Background Technology

[0002] Stroke (including acute ischemic stroke and cerebral hemorrhage) leads to a local inflammatory response caused by factors such as neuronal death and the release of damage-associated molecular patterns (DAMPS). This focal inflammation contributes to secondary damage by exacerbating blood-brain barrier impairment, microvascular failure, cerebral edema, oxidative stress, and directly inducing neuronal death. Furthermore, in addition to local responses in the injured area, increasing evidence suggests that post-stroke inflammatory responses occur and persist throughout the entire brain. Global brain inflammation may continue to influence post-stroke pathological changes and long-term neurological prognosis; understanding the mechanisms regulating global brain inflammation will help in identifying potential treatment strategies.

[0003] Research on pathological interventions for ischemic injury mainly focuses on improving cerebral blood circulation and neuroprotection. Among these, the primary measure for improving cerebral blood circulation is antithrombotic therapy. Antithrombotic drugs, based on their mechanisms of action, can be divided into thrombolytic agents, antiplatelet aggregation agents, and anticoagulants. However, clinical trials have shown that all existing antithrombotic drugs have certain limitations and side effects in clinical application. For example, while anticoagulants can limit the embolism and development of venous and arterial thrombosis, they are less effective in preventing platelet-dominated arterial thrombosis; thrombolytic drugs require sophisticated purification techniques, and the products may be antigenic, potentially inducing allergic reactions; some products can also interfere with coagulation function, posing a risk of bleeding; antiplatelet aggregation agents have high activity and relatively few side effects, but they also carry the risk of delayed bleeding.

[0004] Free radicals are products of normal cellular aerobic metabolism. Highly reactive, they can react with biomolecules (proteins, lipids, sugars, DNA), altering their structure and function. Under normal circumstances, the body has a complete antioxidant system to maintain free radical metabolic balance. However, in pathological conditions (such as stroke, diabetes, aging, etc.), excessive production or impaired elimination of free radicals can lead to lipid peroxidation damage to biological membranes and macromolecules, triggering pathological oxidative stress responses in the body.

[0005] The brain is one of the organs with the highest oxygen load and most active metabolism in the body. Compared with other organs, brain tissue is more prone to producing free radicals and lipid peroxides, and is more susceptible to oxidative stress. Free radical attacks on nerve cells can lead to degenerative changes and neurodegenerative diseases such as Parkinson's disease (PD), Alzheimer's disease (AD), multiple sclerosis (MS), and amyotrophic lateral sclerosis (ALS); it can also be used to treat diabetic peripheral neuropathy (DPN) and diabetic retinopathy (DR) caused by microvascular and nerve damage in diabetes. Oxidative stress is involved in the entire process of neurological diseases such as ischemic cerebrovascular disease, from pathological initiation to prognostic recovery. Inhibiting oxidative stress and scavenging free radicals are important treatment strategies throughout the course of ischemic cerebrovascular disease. The use of antioxidants at different stages of ischemic cerebrovascular disease can bring different benefits to patients and plays an important role in preventing and controlling the occurrence and development of cerebrovascular diseases.

[0006] Currently, there are limited therapeutic drugs available in this field, and no new products have been launched for many years. While this type of disease poses a significant threat, its pathogenesis remains unclear. Based on the limited understanding of this area, treatment strategies that address both the underlying causes and symptoms have gained unanimous approval from experts. Therefore, designing and developing stroke drugs with both neuroprotective and thrombolytic mechanisms of action is of great importance. Summary of the Invention

[0007] The present invention provides a pharmaceutical composition comprising a compound of formula I and dextroborneol. When used in combination, this pharmaceutical composition can significantly exert neuroprotective and thrombolytic therapeutic effects, achieving unexpected therapeutic results.

[0008] This invention provides a pharmaceutical composition that can be more stably stored after being prepared into a formulation.

[0009] This invention provides a pharmaceutical composition that is uniformly distributed in the body and can smoothly cross the blood-brain barrier to act on cerebral blood vessels. The dosage is reduced but the therapeutic effect is not affected, and the side effects on the body are reduced.

[0010] This invention relates to a compound of formula I and dextromethorphanol.

[0011]

[0012] Where R0 is H, Na, or K;

[0013] X is 0, 0.5, 1, 1.5, 2, 2.5, or 3.

[0014] The pharmaceutical composition, specifically, is a compound of formula I, where R0 is selected from Na or K.

[0015] The pharmaceutical composition is characterized in that the dextroborneol is a pharmaceutically available isomer of natural borneol, (+)-2-borneol and its enantiomers, diastereomers, (-)-2-borneol and its enantiomers, diastereomers, racemates, etc.; preferably (+)-2-borneol.

[0016] The pharmaceutical composition comprises a compound structure of Formula I and dextroborneol, characterized in that the weight ratio of the compound structure of Formula I to dextroborneol is 1:20 to 20:1.

[0017] The pharmaceutical composition comprises multiple dosage forms, characterized in that it can be multiple dosage forms, including tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, capsules, oral liquids, lozenges, granules, powders, pills, ointments, elixirs, suspensions, powders, injections, suppositories, creams, sprays, drops, or patches, preferably injections.

[0018] The pharmaceutical composition is characterized by further comprising a pharmaceutically acceptable solvent.

[0019] The pharmaceutical composition is characterized in that the solvent is water, or a water-soluble organic solvent, or a mixture of a water-soluble organic solvent and water.

[0020] The pharmaceutical composition is characterized in that the pharmaceutical carrier comprises one or more of an isotonic agent, an antioxidant, a pH adjuster, a solubilizer, or a cosolvent.

[0021] The pharmaceutical composition is characterized in that the solubilizer is selected from one or more of propylene glycol, ethanol, polyethylene glycol, Tween, and glycerin.

[0022] The pharmaceutical composition is characterized in that the isotonic agent is selected from one or more of sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium lactate, glucose, sorbitol, xylitol, or dextran.

[0023] The pharmaceutical composition is characterized in that the antioxidant is selected from one or more of sodium sulfite, sodium metabisulfite, sodium bisulfite, sodium thiosulfate, or vitamin C.

[0024] The use of the pharmaceutical composition in the preparation of a medicament for treating diseases caused by free radical-mediated oxidative stress and / or thrombosis.

[0025] The aforementioned use is characterized in that the diseases caused by the free radical-mediated oxidative stress and / or thrombosis include acute stroke, arteriosclerosis, Alzheimer's disease, Parkinson's disease, multiple sclerosis, myocardial ischemia and ischemia-reperfusion injury, myocardial infarction, coronary heart disease, or amyotrophic lateral sclerosis.

[0026] In this invention, for a more detailed understanding of the invention, the terms are defined as follows.

[0027] In this invention, the term "subject of treatment" refers to mammals, such as mice, rats, monkeys, dogs, pigs, rabbits, and humans.

[0028] The term "relief and / or treatment" in this invention refers to the effect of reducing or even reversing the condition of a subject or individual by administering an effective therapeutic amount of a compound or its composition to the subject or individual in need of treatment.

[0029] In this invention, the term "treatment" refers to actions taken to inhibit the progression of an applicable disorder or one or more conditions, or to reverse symptoms, and also includes adjunctive treatment for conditions. Detailed Implementation

[0030] The present invention is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the present invention.

[0031] AIBN: Azobisisobutyronitrile

[0032] NBS: N-bromosuccinimide

[0033] DMF: N,N-dimethylformamide

[0034] EA: Ethyl acetate

[0035] Example 1: Synthesis of Compound III

[0036] Reaction formula:

[0037]

[0038] Preparation method:

[0039] Step 1: Synthesis of Compound 2

[0040] Ligustrazine (compound 1, 10.0 g, 73.48 mmol) was dissolved in carbon tetrachloride (100 mL), and AIBN (0.1 g) was added. The mixture was heated to 60 °C, and NBS (15.7 g, 88.17 mmol) was slowly added. After the addition was complete, the temperature was maintained for 18 hours. After the reaction was complete, the system was cooled and poured into ice water. The mixture was extracted three times with methyl tert-butyl ether, and the liquid phases were separated. The organic phases were combined, dried, concentrated, and subjected to column chromatography to give compound 2: 11.7 g, yield: 74.52%.

[0041] Step 2: Synthesis of Compound 4

[0042] Compound 2 (10.0 g, 46.72 mmol) and compound 3 (10.9 g, 49.06 mmol) were mixed and dissolved in DMF (100 mL). Potassium carbonate (8.3 g, 60.46 mmol) was added, and the mixture was stirred at room temperature for 12 hours. After the reaction was complete, the mixture was poured into ice water, extracted three times with EA, and the organic phases were separated. The combined organic phases were washed with saturated brine, dried, concentrated, and subjected to column chromatography to obtain compound 4: 14.3 g, yield: 86.30%.

[0043] 1 HNMR(400 MHz):7.62(d,1H,J=15.9Hz),7.06(m,3H),6.31(d,1H,J=15.9Hz),5.24(s,2H),4.2 6(q,2H,J=7.11Hz),3.82(s,3H),2.65(s,3H),2.52(s,6H),1.34(t,3H,J=7.11Hz).

[0044] Step 3: Synthesis of Compound III

[0045] Compound 4 (10.0 g, 28.08 mmol) was dissolved in ethanol (100 mL), and 50% sodium hydroxide aqueous solution (50 mL) was added at room temperature. After the addition was complete, the system was heated to 60 °C and reacted for 6 hours. After the reaction was complete, the system was cooled to room temperature, concentrated to remove the solvent, and the pH was adjusted to approximately 3 with 2N HCl. A large amount of solid precipitated out. The solid was filtered, and the filter cake was washed with cold water and dried to obtain compound III: 8.3 g, yield: 90.12%.

[0046] 1 HNMR(400 MHz):7.62(d,1H,J=15.9Hz),7.06(m,3H),6.31(d,1H,J=15.9Hz),5.24(s,2H),3.82(s,3H),2.65(s,3H),2.52(s,6H).

[0047] MS (M+1): 329.16.

[0048] Example 2: Synthesis of Compound IV

[0049] Reaction formula:

[0050]

[0051] Preparation method:

[0052] Compound III (5.0 g, 15.24 mmol) was dissolved in anhydrous ethanol (50 mL), and sodium hydroxide (0.6 g, 15.24 mmol) was added. The mixture was stirred at room temperature for 18 hours. After the reaction was complete, a large amount of solid precipitated out. The solid was filtered, washed with cold anhydrous ethanol, and dried to give compound IV: 5.0 g, yield: 93.81%.

[0053] 1 HNMR(400 MHz):7.62(d,1H,J=15.9Hz),7.06(m,3H),6.31(d,1H,J=15.9Hz),5.24(s,2H),4.2 6(q,2H,J=7.11Hz),3.82(s,3H),2.65(s,3H),2.52(s,6H),1.34(t,3H,J=7.11Hz).

[0054] MS (M+1): 329.16.

[0055] Example 3: Synthesis of Compound V

[0056] Reaction formula:

[0057]

[0058] Preparation method:

[0059] Compound III (5.0 g, 15.24 mmol) was dissolved in anhydrous ethanol (50 mL), and potassium hydroxide (0.9 g, 15.24 mmol) was added. The mixture was stirred at room temperature for 18 hours. After the reaction was complete, a large amount of solid precipitated out. The solid was filtered, washed with cold anhydrous ethanol, and dried to give compound IV: 5.0 g, yield: 89.61%.

[0060] 1 HNMR(400 MHz):7.62(d,1H,J=15.9Hz),7.06(m,3H),6.31(d,1H,J=15.9Hz),5.24(s,2H),4.2 6(q,2H,J=7.11Hz),3.82(s,3H),2.65(s,3H),2.52(s,6H),1.34(t,3H,J=7.11Hz).

[0061] MS (M+1): 329.16.

[0062] Example 4: Preparation of Composition 1

[0063]

[0064] Take 1g of dextromethorphanol and 2g of compound III, respectively, and add them to 200ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonic adjuster. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0065] Example 5: Preparation of Composition 2

[0066]

[0067] Take 1g of dextromethorphanol and 3g of compound III, add them to 100ml of ethanol solution, stir to dissolve, then slowly add almost the full volume of water for injection to dissolve the isotonic adjuster, and adjust to the appropriate pH value using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0068] Example 6: Preparation of Composition 3

[0069]

[0070] Take 1g of dextromethorphanol and 4g of compound III, respectively, and add them to 200ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0071] Example 7: Preparation of Composition 4

[0072]

[0073] Take 1g of dextromethorphanol and 5g of compound III, add them to 200ml of propylene glycol solution, stir to dissolve, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant, and adjust the pH value to a suitable value using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0074] Example 8: Preparation of Composition 5

[0075]

[0076] Take 1g of dextromethorphanol and 6g of compound III, respectively, and add them to 150ml of glycerol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0077] Example 9: Preparation of Composition 6

[0078]

[0079] Take 1g of dextromethorphanol and 2g of compound IV, respectively, and add them to 150ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0080] Example 10: Preparation of Composition 7

[0081]

[0082] Take 1g of dextromethorphanol and 3g of compound IV, respectively, and add them to 150ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0083] Example 11: Preparation of Composition 8

[0084]

[0085] Take 1g of dextromethorphanol and 4g of compound IV, respectively, and add them to 150ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0086] Example 12: Preparation of Composition 9

[0087]

[0088] Take 0.5g of dextromethorphanol and 2.5g of compound IV, respectively, and add them to 80ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0089] Example 13: Preparation of Composition 10

[0090]

[0091] Take 1g of dextromethorphanol and 6g of compound IV, respectively, and add them to 150ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0092] Example 14: Preparation of Composition 11

[0093]

[0094] Take 1g of dextromethorphanol and 2g of compound V, respectively, and add them to 150ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0095] Example 15: Preparation of Composition 12

[0096]

[0097] Take 1g of dextromethorphanol and 3g of compound V, respectively, and add them to 150ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0098] Example 16: Preparation of Composition 13

[0099]

[0100] Take 1g of dextromethorphanol and 4g of compound V, respectively, and add them to 150ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0101] Example 17: Preparation of Composition 14

[0102]

[0103] Take 1g of dextromethorphanol and 5g of compound V, respectively, and add them to 150ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0104] Example 18: Preparation of Composition 15

[0105]

[0106] Take 1g of dextromethorphanol and 6g of compound V, respectively, and add them to 150ml of propylene glycol solution. Stir to dissolve them, then slowly add almost the full volume of water for injection to dissolve the isotonicity regulator and antioxidant. Adjust the pH value to a suitable level using a pH adjuster, bring the volume to 1000ml, filter twice, fill and sterilize to obtain the final product.

[0107] Example 19: Preparation of Composition 16

[0108]

[0109] (1) Take the prescribed amount of povidone and add it to an appropriate amount of ethanol solution, and stir to disperse and dissolve it.

[0110] (2) Dissolve the prescribed amount of dextromethorphan (0.5g) and compound V (2.5g).

[0111] (3) Weigh out the prescribed amounts of microcrystalline cellulose, lactose, and crospovidone, and mix them evenly according to the principle of increasing the amount in equal increments.

[0112] (4) Add the solution from (2) to (3) for granulation. Use 24 mesh granulation and dry at 45℃. After drying, granulate to 20 mesh.

[0113] (5) Add stearic acid to the granulated material in (4) and mix evenly. When the final mixture is obtained, compress it into tablets according to the theoretical amount and coat it with a gastric-soluble film coating premix. Finally, the aluminum-plastic sample is prepared.

[0114] Example 20: Preparation of Composition 17

[0115]

[0116] (1) Take all raw and auxiliary materials except magnesium stearate, weigh them according to the prescription amount, and mix them evenly according to the principle of equal increment.

[0117] (2) The dry granulation process is used for granulation, with 24-mesh granulation and 18-mesh granulation.

[0118] (3) Add the prescribed amount of magnesium stearate to the granulated material and mix well.

[0119] (4) Compress the tablets according to the theoretical amount, and then make the final product from aluminum and plastic.

[0120] Example 21: Formulation of Composition 18

[0121]

[0122] (1) Weigh out the prescribed amount of propylene glycol, add compound V and dextroborneol to dissolve, and then add the prescribed amount of sorbitol solution to dissolve.

[0123] (2) Measure 400ml of water, add the prescribed amount of hydroxypropyl methylcellulose, sorbic acid and flavoring and dissolve completely.

[0124] (3) Add the solution in 2 to (1) until completely dissolved and bring the volume up to 1000ml. Fill the solution into an oral liquid bottle to obtain the finished product.

[0125] Example 22: Rat Cerebral Infarction Experiment

[0126] Selection of experimental animals:

[0127] Eighty healthy male SPF-grade SD rats weighing 200-230g were randomly selected and housed in an environment of 23±2℃ with 12 hours of alternating light and dark conditions. During the housing process, the rats had free access to food and water and were allowed to adapt for one week.

[0128] After acclimatization, the rats were divided into 10 groups of 8 rats each, for a total of 80 rats, based on their body weight.

[0129] Preparation and administration of the test drug:

[0130] Preparation of piradogrel: Weigh 33.75 mg of piradogrel, dissolve it in an appropriate amount of DMSO, and then make up to 100 mL with physiological saline. Shake well to mix thoroughly to obtain the piradogrel solution.

[0131] Preparation of dexborneol: Weigh 11.25 mg of dexborneol, dissolve it in an appropriate amount of DMSO, and make up to 100 mL with physiological saline. Shake well to mix thoroughly to obtain the dexborneol solution.

[0132] Preparation of positive control drug: Accurately measure 5.40 mL of concentrated edaravone dexborneol injection solution (Syntox, ED, Sinopharm Co., Ltd.), add an appropriate amount of DMSO to dissolve, and then make up to 40 mL with physiological saline. Shake well to mix thoroughly to obtain the positive control drug solution.

[0133] Preparation of Composition 1: Accurately measure 4.500 mL of Composition 1, add an appropriate amount of DMSO to dissolve it, and then make up to 40 mL with physiological saline. Shake well to mix thoroughly to obtain the solution of Composition 1.

[0134] Preparation of Composition 3: Accurately measure 2.700 mL of Composition 3, add an appropriate amount of DMSO to dissolve it, and then make up to 40 mL with physiological saline. Shake well to mix thoroughly to obtain the solution of Composition 3.

[0135] Preparation of Composition 5: Accurately measure 1.929 mL of Composition 5, add an appropriate amount of DMSO to dissolve it, and then make up to 40 mL with physiological saline. Shake well to mix thoroughly to obtain the solution of Composition 5.

[0136] Preparation of Composition 8: Accurately measure 2.700 mL of Composition 8, add an appropriate amount of DMSO to dissolve it, and then make up to 40 mL with physiological saline. Shake well to mix thoroughly to obtain the solution of Composition 8.

[0137] Preparation of Composition 13: Accurately measure 2.700 mL of Composition 13, dissolve it in an appropriate amount of DMSO, and then dilute to 40 mL with physiological saline. Shake thoroughly to mix well to obtain the Composition 13 solution. All solutions should be prepared and used immediately.

[0138] Administration method of the test drug: see Table 19.

[0139]

[0140] Test method:

[0141] After the rats' adaptation period, behavioral training (balance beam, rotisserie) was initiated three days prior to surgery for all rats in each group. Rats were fasted for 12 hours preoperatively but allowed free access to water. Before surgery, all rats were anesthetized with isoflurane, maintaining spontaneous respiration. They were fixed in a supine position on a rat board, with hair removed from the midline of the neck and disinfected with 75% alcohol. A surgical incision was made on the ventral side of the rat's neck in the midline. The muscle and fascia were separated along the inner edge of the sternocleidomastoid muscle, and the right common carotid artery, external carotid artery, and internal carotid artery were isolated. A small oblique incision was made on the external carotid artery approximately 0.5 cm proximal to the ligation site using vascular scissors. The external carotid artery was pulled until it was aligned with the internal carotid artery. The suture occluded the rat 1.8 cm slowly through the incision in the right external carotid artery towards the intracranial entry point of the internal carotid artery, using the bifurcation of the common carotid artery as a marker. Slight resistance was felt during advancement, indicating that the middle cerebral artery had been blocked. The suture occluded 2 hours after infarction, completing the cerebral ischemia-reperfusion injury model. The sham-operated group only underwent vascular dissection. Twenty minutes after infarction, cerebral blood flow was assessed using Doppler flowmetry. A model was considered successful and included in the experiment if the difference in blood flow between the left and right hemispheres (ROI%) was greater than 48%. Rectal temperature was maintained intraoperatively using an incandescent lamp. Postoperatively, medication was administered according to the settings in the table above. Drugs for each group were administered intravenously 0.5 hours after cerebral infarction. The sham-operated group and the model group received the same volume of blank solvent.

[0142] Twenty-four hours after reperfusion, the Zea-longa neurological function score was assessed for each animal. At the end of the experiment (24 hours after reperfusion), changes in cerebral blood flow were measured using Doppler flowmeters to evaluate the ameliorative effect of the test substance on ischemic stroke. Behavioral function tests were also conducted. One hour later, all animals were euthanized, and whole brain tissue was harvested and weighed to determine the wet weight of the brain. Brain volume was measured using the water displacement method. Serial coronal sections of the whole brain were prepared and stained with TTC. The infarct area was measured using Image-J software, and the percentage of the infarct area to the total brain area was calculated. The infarct improvement rate was also calculated.

[0143] Zea-Longa scoring criteria: No neurological deficits: 0 points; Inability to fully extend the forepaw on the paralyzed side: 1 point; Turning in circles towards the paralyzed side while walking: 2 points; Leaning towards the paralyzed side while walking: 3 points; Inability to walk automatically, with signs of loss of consciousness: 4 points.

[0144] Infarct area ratio = Total infarct area ÷ Total area of ​​whole brain slices × 100%;

[0145] Infarction improvement rate = (infarction area ratio in the model group - infarction area ratio in the treatment group) ÷ infarction area ratio in the model group × 100%.

[0146] SPSS 21.0 software was used for the corresponding statistical analysis. A p < 0.05 was considered statistically significant, and a p < 0.01 was considered highly statistically significant.

[0147] The evaluation results of the neurological function of rats in each group are shown in Table 20.

[0148]

[0149]

[0150] The results of behavioral function tests of rats in each group are shown in Table 21.

[0151]

[0152]

[0153] The maximum time for the balance beam test is 60 seconds. If the test exceeds 60 seconds, it will be treated as 60 seconds.

[0154] The results of the wet weight test of the brains of rats in each group are shown in Table 22.

[0155]

[0156]

[0157] The brain volume data of rats in each group are shown in Table 23.

[0158]

[0159]

[0160] The data on cerebral infarction area and infarction improvement rate of each group of rats are shown in Table 24.

[0161]

[0162]

[0163] Experimental conclusions: The above experimental data show that, compared with the model group, the other groups have significant improvement effects on the core evaluation indicators of neurological function impairment (neurological function, behavioral function, brain wet weight, brain volume, and cerebral infarction area and infarction improvement rate) in rats with acute ischemic stroke. Compositions 1 and 5 are slightly better than the two single-drug groups (bilagrelor group and dexborneol group) and the positive control group, especially compositions 3, 8, and 13. In terms of cerebral infarction area and infarction improvement rate, the compositions disclosed in this invention are better than the two single-drug groups (bilagrelor and dexborneol) and the positive control group, demonstrating excellent neuroprotective ability.

[0164] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A pharmaceutical composition wherein the active ingredient comprises a compound of formula I and dextromethorphanol. in: R0 is H, Na, or K; X is 0, 0.5, 1, 1.5, 2, 2.5, or 3.

2. The pharmaceutical composition according to claim 1, particularly the compound of formula I, wherein RO is selected from Na or K.

3. The pharmaceutical composition according to claim 1, characterized in that, The dextroborneol is a pharmaceutically available isomer of natural borneol, (+)-2-camphenol and its enantiomers, diastereomers, (-)-2-camphenol and its enantiomers, diastereomers, racemates, etc.; preferably (+)-2-camphenol.

4. The pharmaceutical composition according to claims 1-3, characterized in that, The weight ratio of the compound of Formula I to dextromethorphan is 1:20 to 20:

1.

5. The pharmaceutical composition according to any one of claims 1 to 4, characterized in that, It can be a variety of drug dosage forms, including tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, capsules, oral liquids, lozenges, granules, powders, pills, powders, ointments, elixirs, suspensions, powders, injections, suppositories, creams, sprays, drops, or patches, preferably injections.

6. The pharmaceutical composition according to claim 5, characterized in that, It also includes pharmaceutically acceptable solvents.

7. The pharmaceutical composition according to claim 6, characterized in that, The solvent is water, or a water-soluble organic solvent, or a mixture of a water-soluble organic solvent and water.

8. The pharmaceutical composition according to claim 5 or 7, characterized in that, The pharmaceutical carrier includes one or more of the following: isotonic agents, antioxidants, pH adjusters, solubilizers, or cosolvents.

9. The pharmaceutical composition according to claim 8, characterized in that, The solubilizer is selected from one or more of propylene glycol, ethanol, polyethylene glycol, Tween, and glycerin.

10. The pharmaceutical composition according to claim 8 or 9, characterized in that, The isotonic agent is selected from one or more of sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium lactate, glucose, sorbitol, xylitol, or dextran.

11. The pharmaceutical composition according to any one of claims 8-10, characterized in that, The antioxidant is selected from one or more of sodium sulfite, sodium metabisulfite, sodium bisulfite, sodium thiosulfate, or vitamin C.

12. Use of the pharmaceutical composition according to any one of claims 1 to 11 in the preparation of a medicament for treating diseases caused by free radical-mediated oxidative stress and / or thrombosis.

13. The use according to claim 12, characterized in that, Diseases caused by free radical-mediated oxidative stress and / or thrombosis include acute stroke, arteriosclerosis, Alzheimer's disease, Parkinson's disease, multiple sclerosis, myocardial ischemia and ischemia-reperfusion injury, myocardial infarction, coronary heart disease, or amyotrophic lateral sclerosis.

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