Pharmaceutical composition and application

Abstract

The invention discloses a pharmaceutical composition and application, and the pharmaceutical composition is prepared from the following components in parts by weight: 1 part by weight of 2-hydroxy-6-trifluoromethyl dexcamphyl nicotinate and 4-19 parts by weight of cilostazol or riluzole. According to the invention, the medicine synergistic effect of the 2-hydroxy-6-trifluoromethyl nicotinic acid dexcamphanol ester and cilostazol or riluzole is proposed and verified for the first time, and the combined medication effect strength of the most preferable formula can reach 1.30; besides, the pharmaceutical composition can effectively reduce the cerebral infarction range of a focal cerebral ischemia reperfusion model and improve the trend of cerebral ischemia injury, and has great clinical application prospects.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical chemistry, in particular to a pharmaceutical composition and application. BACKGROUND

[0002] Cerebral vascular disease (CVD) refers to brain dysfunction caused by cerebral vascular disease, which is mainly divided into acute and chronic types. Ischemic stroke, as the core type of acute CVD, is a critical disease caused by stenosis or occlusion of cerebral blood vessels, including carotid and vertebral arteries, leading to ischemic necrosis of brain tissue, accounting for a large proportion of all strokes, and timely intervention is crucial to improve the prognosis.

[0003] As a selective PDE3 inhibitor, cilostazol plays an important role in the prevention and treatment of cerebral ischemic diseases. Clinical studies have confirmed that cilostazol not only delays the cognitive decline of cerebral small vessel disease (CSVD) patients, such as significantly improving the MoCA score, but also improves the gait speed and stability of patients through combination with balance training, providing a comprehensive management strategy for CSVD, an important brain ischemia-related disease. Riluzole is another promising candidate compound for the prevention and treatment of cerebral ischemic diseases. Although it is famous for treating amyotrophic lateral sclerosis (ALS), many studies have shown that it has potential neuroprotective ability. The core role of riluzole is to inhibit the excessive release of glutamate in pathological conditions and partially block NMDA receptors, thereby reducing excitotoxicity damage. Middle cerebral artery occlusion model (MCAO) shows that early administration of riluzole after ischemia can effectively reduce the neurological deficit score and reduce the infarct area. Its mechanism also involves anticonvulsant and antioxidant effects, providing a new idea for stroke neuroprotection. Although clinical translation is still under exploration, the evidence from basic research is clear. In addition, the invention patent CN119504571A discloses a class of 2-hydroxy nicotinic acid d-camphol or fenchol ester derivatives, specifically discloses 2-hydroxy-6-trifluoromethyl nicotinic acid d-camphol ester, which has various biological activities, such as anti-inflammatory, analgesic, and reducing nerve damage after stroke.

[0004] However, there is currently no report on the combination of the above compounds. SUMMARY

[0005] The purpose of the present application is to provide a pharmaceutical composition based on 2-hydroxy-6-trifluoromethyl nicotinic acid d-camphol ester, containing cilostazol or riluzole, and the second purpose is to provide the application of the pharmaceutical composition.

[0006] The pharmaceutical composition of the present application contains 1 part by weight of 2-hydroxy-6-trifluoromethyl nicotinic acid d-camphol ester, and 4-19 parts by weight of cilostazol or riluzole.

[0007] Preferably, the pharmaceutical composition contains 1 part by weight of 2-hydroxy-6-trifluoromethyl nicotinic acid dexrazoxane ester, and 8-10 parts by weight of cilostazol or riluzole.

[0008] Preferably, the pharmaceutical composition further contains a pharmaceutically acceptable adjuvant.

[0009] Preferably, the dosage form of the pharmaceutical composition is granules, tablets, capsules, oral liquids, pills, emulsions, suspensions, injections or infusions.

[0010] The pharmaceutical composition of the present application is used in the preparation of a drug for treating cerebrovascular diseases.

[0011] Preferably, the cerebrovascular disease is cerebral ischemic disease.

[0012] Preferably, the cerebral ischemic disease is ischemic stroke.

[0013] Preferably, the drug is an intravenous drug.

[0014] Preferably, the use is the use in the preparation of a drug for improving cerebral ischemic injury.

[0015] Preferably, the drug is a drug for reducing the range of cerebral infarction.

[0016] Beneficial effects: Compared with the prior art, the present application has the following remarkable advantages: 1. The present application first proposes and verifies the drug synergistic effect of 2-hydroxy-6-trifluoromethyl nicotinic acid dexrazoxane ester and cilostazol or riluzole, and the combined use of the most preferred formula can reach 1.30 in intensity; 2. The combined use of 2-hydroxy-6-trifluoromethyl nicotinic acid dexrazoxane ester and cilostazol or riluzole can effectively reduce the range of cerebral infarction in the focal cerebral ischemia reperfusion model, improve the trend of cerebral ischemic injury, and has great clinical application prospect. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 Statistical diagram of the influence of 2-hydroxy-6-trifluoromethyl nicotinic acid dexrazoxane ester with different proportions of cilostazol on the range of cerebral infarction at a dose of 1 mg / kg;

[0018] Figure 2 Statistical diagram of the influence of 2-hydroxy-6-trifluoromethyl nicotinic acid dexrazoxane ester with cilostazol on the range of cerebral infarction at a total dose of 10 mg / kg;

[0019] Figure 3 Statistical diagram of the influence of 2-hydroxy-6-trifluoromethyl nicotinic acid dexrazoxane ester with riluzole on the range of cerebral infarction at a dose of 1 mg / kg;

[0020] Figure 4Statistical chart of the influence of the compound use of 2-hydroxy-6-trifluoromethyl nicotinic acid dexpanthenol ester and riluzole with a total dose of 10 mg / kg on the scope of cerebral infarction. DETAILED DESCRIPTION

[0021] The technical solutions of the present application are further described below.

[0022] Example 1: 2-hydroxy-6-trifluoromethyl nicotinic acid dexpanthenol ester (LL-06) and cilostazol combination therapy for ischemic stroke

[0023] 1. 2-hydroxy-6-trifluoromethyl nicotinic acid dexpanthenol ester at a dose of 1 mg / kg combined with cilostazol at different ratios

[0024] (1) SPF level male Sprague Dawley (SD) rats weighing 250-280 g were used to prepare a rat focal cerebral ischemia reperfusion model by the method of internal carotid suture: the rats were fixed in a supine position on the operating table after being anesthetized, the skin was alcohol disinfected after shaving from the head end to the chest; make an incision in the middle of the neck, bluntly dissect the subcutaneous tissue, separate the thin layer of fascia on the surface of the anterior triangle of the neck, push up the lower edge of the hyoid bone-laryngeal muscle, open the arterial shell, expose the right carotid bifurcation, separate the right common carotid artery, external carotid artery, and internal carotid artery, ligate and cut the external carotid artery; clamp the proximal end of the common carotid artery, make an incision at the distal end of the ligation line of the external carotid artery, insert the suture, pass through the common carotid artery bifurcation into the internal carotid artery, and block all blood supply to the middle cerebral artery; loosen the proximal end of the common carotid artery clamp, after 2 hours of right cerebral ischemia, pull out the suture, restore blood supply for reperfusion, ligate the external carotid artery, suture the skin, and complete the operation;

[0025] (2) After the preparation of the focal cerebral ischemia reperfusion model, the rats were single-blindly assigned to: 1 mg / kg LL-06 group, 4 mg / kg cilostazol group, 9 mg / kg cilostazol group, 19 mg / kg cilostazol group, composition 1 group-4 mg / kg cilostazol + 1 mg / kg LL-06 (cilostazol: LL-06 = 4:1), composition 2 group-9 mg / kg cilostazol + 1 mg / kg LL-06 (cilostazol: LL-06 = 9:1), composition 3 group-19 mg / kg cilostazol + 1 mg / kg LL-06 (cilostazol: LL-06 = 19:1), and model group, a total of 8 groups, 8 rats in each group, intravenous administration once immediately, and the model group animals were given an equal volume of normal saline;

[0026] (3) After 24 hours, the brain was euthanized with CO2. The olfactory bulb, cerebellum and lower brainstem were removed. The surface of the brain was rinsed with physiological saline to remove bloodstains and residual water. The brain was placed at -20℃ for 20 minutes. Immediately after removal, a coronal section was made vertically downward at the plane of the intersection of the lines of sight, and slices were cut every 2 mm backward. The brain slices were placed in 1% triphenyltetrazolium chloride (TTC) staining solution and incubated at 37℃ for 30 minutes. After rinsing with physiological saline, the brain slices were arranged in front to back and photographed.

[0027] The obtained images were processed using ImageJ software. Dark red represented normal brain tissue, and pale white represented ischemic brain tissue. The specific area of ​​different brain tissues was quantified. The volume of the left brain and the volume of the non-infarcted area of ​​the right brain were calculated according to formula (I), and the percentage of cerebral infarction area (%I) was further calculated according to formula (II).

[0028] V = t(A1 + A2 + A3 + ... + A n (I),

[0029] Where t is the slice thickness and An is the non-cerebral infarction area.

[0030] %I = 100% × (V) C -V L ) / V C (II),

[0031] Among them, V C V represents the brain volume of the control side (left hemisphere). L The volume of the non-infarcted region on the infarcted side (right hemisphere).

[0032] The percentage of cerebral infarction area is expressed as mean ± standard deviation (Mean ± SD). One-way ANOVA was used to analyze differences between groups, and the LSD test was used for inter-group comparisons. A p-value < 0.05 was defined as statistically significant.

[0033] (4) Evaluate the combined pharmacodynamic effect q of LL-06 and cilostazol in the composition according to Jin Zhengjun's formula (III):

[0034] q = E a+b / (E a +E b E a ×E b (III),

[0035] Among them, E a+b The efficacy rates of the combined medications, Ea and Eb, are the efficacy rates of LL-06 and cilostazol as monotherapy, respectively, calculated according to formula (IV):

[0036] E 给药组 =(X 模型组 -X给药组 ) / X 模型组 (IV),

[0037] wherein, X 模型组 is the percentage of cerebral infarction area of the model group, and X 给药组 is the percentage of cerebral infarction area of the administration group.

[0038] The effect of the administration on the cerebral infarction area is shown in Figure 1 and Table 1. The experimental results show that 1 mg / kg LL-06, 9 mg / kg cilostazol, 19 mg / kg cilostazol, or the combination of LL-06 and cilostazol can significantly reduce the cerebral infarction area of the rat focal cerebral ischemia-reperfusion model.

[0039] According to the Jin Zhengjun formula, if the effect intensity q value of the combination is in the range of 0.85-1.15, it is simple addition of the two drugs, q value > 1.15 is synergy, and q value < 0.85 indicates antagonism of the combination of the two drugs. The q value of the combination of LL-06 and cilostazol is > 1.15, indicating that the combination of the two drugs has a synergistic effect.

[0040] Table 1. Effect of the combination of cilostazol and LL-06 on the cerebral infarction area (fixed LL-06 dose)

[0041] Group Cerebral infarction range (%) Synergy (q) Model group 38.81±7.34 / 1 mg / kg LL-06 group 28.76±7.06** / 4 mg / kg cilostazol group 32.78±5.74 / 9 mg / kg cilostazol group 30.21±7.45* / 19 mg / kg cilostazol group 27.18±7.52** / Cilostazol: LL-06 = 4:1 group 21.38±9.47*** 1.20 Cilostazol: LL-06 = 9:1 group 17.66±5.86*** 1.29 Cilostazol: LL-06 = 19:1 group 16.37±6.39*** 1.20

[0042] wherein, ** indicates p < 0.01 compared with the model group, and *** indicates p < 0.001 compared with the model group.

[0043] 2. Combination of 2-hydroxy-6-trifluoromethylnicotinic acid dexpanthenol ester and cilostazol in a total dose of 10 mg / kg for the treatment of ischemic stroke

[0044] According to the foregoing method, SPF male Sprague Dawley (SD) rats weighing 250-280 g were used to prepare a rat focal cerebral ischemia-reperfusion model by the method of internal carotid artery thread embolism. Then the rats were single-blindly assigned to: composition 1 group-8 mg / kg cilostazol + 2 mg / kg LL-06 (4:1), composition 2 group-9 mg / kg cilostazol + 1 mg / kg LL-06 (9:1), composition 3 group-9.5 mg / kg cilostazol + 0.5 mg / kg LL-06 (19:1), and a model group, a total of 4 groups, 12 rats in each group, and intravenous administration was performed once immediately. The animals in the model group were given an equal volume of normal saline.

[0045] According to the foregoing method, the brain was taken out, frozen, sectioned, and stained after 24 h of neurological deficit symptom evaluation, and the percentage of cerebral infarction area and the effect intensity of the combination were calculated according to the foregoing formula.

[0046] The effect of the drug on the cerebral infarction area is shown in Table 2 Figure 2 and Table 2, the experimental results show that the different ratio of LL-06 and cilostazol in the compound drug can significantly reduce the cerebral infarction area of the rat focal cerebral ischemia reperfusion model, and the group of cilostazol: LL-06 = 9: 1 has the best effect.

[0047] Table 2 Effect of cilostazol and LL-06 compound drug on cerebral infarction area (fixed total dose)

[0048] Group Cerebral infarction range (%) Model group 39.04±8.17 Cilostazol: LL-06 = 4:1 group 20.54±7.33*** Cilostazol: LL-06 = 9:1 group 17.42±7.36*** Cilostazol: LL-06 = 19:1 group 22.88±9.51***

[0049] Among them, *** represents p < 0.001 compared with the model group.

[0050] Example 2: 2-hydroxy-6-trifluoromethyl nicotinic acid dexpanthenol ester (LL-06) and riluzole combination therapy for ischemic stroke

[0051] 1, 1mg / kg dose of 2-hydroxy-6-trifluoromethyl nicotinic acid dexpanthenol ester combined with different proportions of riluzole

[0052] According to the foregoing method, SPF male Sprague Dawley (SD) rats weighing 250-280g were used to prepare a rat focal cerebral ischemia reperfusion model by the method of internal carotid artery thread, and then the rats were single-blindly assigned to: 1mg / kg LL-06 group, 4mg / kg riluzole group, 9mg / kg riluzole group, 19mg / kg riluzole group, composition 1 group-4mg / kg riluzole+1mg / kg LL-06 (riluzole: LL-06 = 4:1), composition 2 group-9mg / kg riluzole+1mg / kg LL-06 (riluzole: LL-06 = 9:1), composition 3 group-19mg / kg riluzole+1mg / kg LL-06 (riluzole: LL-06 = 19:1) and model group, a total of 8 groups, 8 rats per group, intravenous administration once immediately, and the model group animals were given an equal volume of normal saline;

[0053] According to the foregoing method, the brain was taken out after 24h, frozen, sectioned, stained, and the percentage of cerebral infarction area and the effect intensity of combined drug were calculated according to the foregoing formula.

[0054] The effect of the drug on the cerebral infarction area is shown in Table 2 Figure 3 and Table 3, the experimental results show that 1mg / kg LL-06, 9mg / kg riluzole, 19mg / kg riluzole or the compound drug of LL-06 and riluzole can significantly reduce the cerebral infarction area of the rat focal cerebral ischemia reperfusion model; at the same time, the q value of the compound drug of LL-06 and riluzole is >1.15, indicating that the combination of the two drugs has a synergistic effect.

[0055] Table 3 Effect of riluzole and LL-06 combination on cerebral infarction size (fixed LL-06 dose)

[0056]

[0057]

[0058] Wherein, * indicates p < 0.05 compared with the model group, ** indicates p < 0.01 compared with the model group, and *** indicates p < 0.001 compared with the model group.

[0059] 2. Combination of 2-hydroxy-6-trifluoromethylnicotinic acid dexpanthenol ester and riluzole in a total dose of 10 mg / kg for treating ischemic stroke

[0060] According to the foregoing method, SPF level male Sprague Dawley (SD) rats weighing 250-280 g were used to prepare a rat focal cerebral ischemia reperfusion model by the internal carotid suture method. Then the rats were single-blindly assigned to: composition 1 group - 8 mg / kg riluzole + 2 mg / kg LL-06 (4:1), composition 2 group - 9 mg / kg riluzole + 1 mg / kg LL-06 (9:1), composition 3 group - 9.5 mg / kg riluzole + 0.5 mg / kg LL-06 (19:1), and a model group, a total of 4 groups, 12 rats in each group, intravenous administration once immediately, and the model group animals were given an equal volume of normal saline.

[0061] According to the foregoing method, the brain was taken out, frozen, sectioned, and stained after 24 h of neurological deficit symptom evaluation, and the percentage of cerebral infarction size and the effect intensity of combined use were calculated according to the foregoing formula.

[0062] The effect of the drug on the size of cerebral infarction is shown in Figure 4 and Table 4, the experimental results show that under the condition of fixed total dose of 10 mg / kg, different ratio of LL-06 and riluzole combination can significantly reduce the size of cerebral infarction in rat focal cerebral ischemia reperfusion model, wherein the riluzole: LL-06 = 9:1 group has the optimal effect.

[0063] Table 4 Effect of riluzole and LL-06 combination on cerebral infarction size (fixed total dose)

[0064] Group Cerebral infarction range (%) Model group 39.23±8.34 Riluzole: LL-06 = 4:1 group 23.88±9.50*** Riluzole: LL-06 = 9:1 group 17.38±8.39*** Riluzole: LL-06 = 19:1 group 22.59±7.12***

[0065] Wherein, *** indicates p < 0.001 compared with the model group.

Claims

1. A pharmaceutical composition, characterized by, The pharmaceutical composition contains 1 part by weight of 2-hydroxy-6-trifluoromethylnicotinic acid dexketoprofen ester and 4 to 19 parts by weight of cilostazol or riluzole.

2. The pharmaceutical composition of claim 1, wherein, The pharmaceutical composition contains 1 part by weight of 2-hydroxy-6-trifluoromethylnicotinic acid dexketoprofen ester and 8 to 10 parts by weight of cilostazol or riluzole.

3. The pharmaceutical composition of claim 1, wherein, The pharmaceutical composition further contains a pharmaceutically acceptable adjuvant.

4. The pharmaceutical composition of claim 1, wherein, The dosage form of the pharmaceutical composition is granules, tablets, capsules, oral solution, pills, emulsion, suspension, injection or infusion.

5. Use of the pharmaceutical composition according to any one of claims 1 to 4 in the manufacture of a medicament for treating cerebrovascular diseases.

6. Use according to claim 5, characterized in that, The cerebrovascular disease is cerebral ischemic disease.

7. Use according to claim 6, characterized in that, The cerebral ischemic disease is ischemic stroke.

8. Use according to claim 5, characterized in that, The medicament is intravenous administration medicament.

9. Use according to claim 5, characterized in that, The use is the use in the manufacture of a medicament for improving cerebral ischemic injury.

10. Use according to claim 9, characterized in that, The medicament is a medicament for reducing the range of cerebral infarction.

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