阅读说明：本技术 Zl006和（+）2-崁醇的组合物及其药物用途 (Composition of ZL006 and (+) 2-anethol and pharmaceutical application thereof ) 是由 李飞 李伦家 徐楠 于 2021-01-28 设计创作，主要内容包括：ZL006和(+)2-崁醇的组合物及其药物用途,其中：ZL006和(+)2-崁醇的质量比为(6～1)：1。ZL006和(+)2-崁醇的组合物,能够显著减少脑梗死面积。与单用ZL006(+)2-崁醇相比,组合物在获得更好药效的同时,其有效剂量显著降低。由于降低了ZL006和(+)2-崁醇的剂量,减少了高剂量可能引起的风险,具有更好的安全性。(A composition of ZL006 and (+) 2-linalool and pharmaceutical uses thereof, wherein: the mass ratio of ZL006 to (+) 2-anethol is (6-1): 1.ZL006 and (+) 2-linalool, significantly reduce cerebral infarct size. The composition has significantly reduced effective dose while achieving better efficacy compared to ZL006(+) 2-chamigol alone. The dosage of ZL006 and (+) 2-linalool is reduced, the risk caused by high dosage is reduced, and the safety is better.)

A composition of ZL006 and (+) 2-linalool, characterized by: the mass ratio of ZL006 to (+) 2-anethol is (6-1): 1.

2. The composition of ZL006 and (+) 2-chamiel as claimed in claim 1, wherein: the mass ratio of ZL006 to (+) 2-anethol is (6-3): 1.

3. The composition of ZL006 and (+) 2-chamiel as claimed in claim 1, wherein: the mass ratio of ZL006 to (+) 2-anethol is (3-1): 1.

4. The composition of ZL006 and (+) 2-chamiel as claimed in claim 1, wherein: the mass ratio of ZL006 to (+) 2-anethol is 3: 1.

5. The composition of ZL006 and (+) 2-chamiel as claimed in claim 1, wherein: ZL0060.84mg/kg and (+) 2-champignon 0.84 mg/kg.

6. Use of a composition of ZL006 and (+) 2-chamielol according to any one of claims 1 to 5 in the manufacture of a medicament for the treatment of stroke.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a composition of ZL006 and (+) 2-anethol and a pharmaceutical application thereof.

Background

Stroke is classified into hemorrhagic stroke and ischemic stroke. Physiologically, glutamate in the central nervous system is mostly localized in presynaptic vesicles, and the content in synaptic cleft is maintained at micromolar level. During cerebral ischemia, the concentration of extracellular glutamic acid is remarkably increased, and Ca is caused by activating N-methyl-D-aspartate receptor (NMDA receptor)²⁺While downstream nNO is activated by the NMDAR/postsynaptic compact protein 95(PSD 95)/neuronal nitric oxide synthase (nNOS) complexS, release of Nitric Oxide (NO), triggering neuronal excitatory injury. The nNOS and NMDA receptors are related to learning and memory ability, and the direct inhibition of the nNOS or NMDA receptors causes side effects such as hypomnesis, mental disorder and the like, so that the interaction between the NMDA receptors and PSD-95 or between the nNOS and PSD-95 is selectively blocked without directly inhibiting the nNOS or NMDA receptors, the release of NO can be inhibited on the premise of not influencing the normal action of the nNOS and NMDA receptors on a central nervous system, and the nerve injury is reduced. 4-N- (2-hydroxy-3, 5-dichlorobenzyl) aminosalicylic acid (ZL006) can specifically block the interaction of PSD-95/nNOS, improve the symptoms of the neural defect of Middle Cerebral Artery Occlusion (MCAO) reperfusion animals and reduce the volume of infarction (Nature Medicine 2010,16, 1439-. Since NO itself has important physiological functions, excessive reduction of NO may cause potential side effects.

(+) 2-anethol has strong inflammation inhibiting effect, and may inhibit the activation of transcription factor NF-kappa B, the expression of inflammatory protein (iNOS and COX-2) and the release of inflammatory cell factor (TNF-alpha, IL-1 beta, etc.) to produce cell protecting effect. High doses of (+) 2-champignon can excite central nerves, cause convulsion, loss of consciousness, spasm and, in severe cases, cause death.

The applicant found that: in the MCAO model: ZL006 and (+) 2-linalool, significantly reduce cerebral infarct size. The composition has significantly reduced effective dose while achieving better efficacy compared to ZL006(+) 2-chamigol alone. The dosage of ZL006 and (+) 2-linalool is reduced, the risk caused by high dosage is reduced, and the safety is better.

Disclosure of Invention

The technical problem to be solved is as follows: the invention provides a composition of ZL006 and (+) 2-linalool and a pharmaceutical application thereof, wherein the composition of ZL006 and (+) 2-linalool has a synergistic effect when being used in a matching way, and the use effect can be obviously improved.

The technical scheme is as follows: the composition of ZL006 and (+) 2-anethol has a mass ratio of ZL006 to (+) 2-anethol of (6-1): 1.

Preferably, the mass ratio of ZL006 to (+) 2-linalool is (6-3): 1.

Preferably, the mass ratio of ZL006 to (+) 2-linalool is (3-1): 1.

It is further preferred that the mass ratio of ZL006 to (+) 2-linalool is 3: 1.

ZL006 and (+) 2-anethol, ZL0060.84mg/kg and (+) 2-anethol 0.84 mg/kg.

The application of the composition of ZL006 and (+) 2-anethol in preparing a medicament for treating cerebral apoplexy diseases.

Has the advantages that: ZL006 and (+) 2-linalool, significantly reduce cerebral infarct size. The composition has significantly reduced effective dose while achieving better efficacy compared to ZL006(+) 2-chamigol alone. The dosage of ZL006 and (+) 2-linalool is reduced, the risk caused by high dosage is reduced, and the safety is better. Therefore, the composition can be used for preparing a medicament for treating cerebral apoplexy.

Drawings

FIG. 1 is a graph of the effect of a combination of ZL006 and (+) 2-linalool on cerebral infarct size in rats (Mean. + -. SEM);

FIG. 2 is a graph of the effect of composition ZL006 and (+) 2-linalool (3: 1) on cerebral infarct size in rats (Mean. + -. SEM).

Detailed Description

The following examples are given to enable a person skilled in the art to fully understand the invention, but do not limit it in any way.

Example 1: preparation of solutions of composition of ZL006 and (+) 2-anethol

1.1 formulation a 1: ZL 00684 mg, (+) 2-anethol 84mg, polyethylene glycol (400)2g, 95% ethanol 1g dissolve, add water for injection and dilute to 100 mL; thus, a solution of a composition (1: 1 by mass) comprising ZL0060.84mg/mL and (+) 2-champignon 0.84mg/mL was obtained.

1.2 formulation a 2: ZL 006126 mg, (+) 2-anethol 42mg, polyethylene glycol (400)2g, 95% ethanol 1g dissolve, add water for injection and dilute to 100 mL; thus, a solution of ZL0061.26mg/mL and (+) 2-champignon 0.42mg/mL (mass ratio: 3:1) was obtained.

1.3 formulation a 3: ZL 006144 mg, (+) 2-anethol 24mg, polyethylene glycol (400)2g, 95% ethanol 1g dissolve, add water for injection and dilute to 100 mL; thus, a solution of ZL0061.44mg/mL and (+) 2-chamanol 0.24mg/mL (mass ratio: 6:1) was obtained.

1.4 formulation a 4: ZL 00663 mg, (+) 2-anethol 21mg, polyethylene glycol (400)2g, 95% ethanol 1g dissolve, add water for injection and dilute to 100 mL; thus, a solution of ZL0060.63mg/mL and (+) 2-champignon 0.21mg/mL (mass ratio: 3:1) was obtained.

1.5 formulation a 5: ZL00631.5 mg, (+) 2-anethol 10.5mg, polyethylene glycol (400)2g, 95% ethanol 1g dissolved, adding water for injection and diluting to 100 mL; thus, a solution of a composition (mass ratio: 3:1) comprising ZL0060.315mg/mL and (+) 2-champignon 0.105mg/mL was obtained.

1.6 formulation B1: ZL 006168 mg, 2g of polyethylene glycol (400) and 1g of 95% ethanol are dissolved, and water for injection is added to dilute the solution to 100 mL; thus, a ZL0061.68mg/mL solution was obtained.

1.7 formulation B2: ZL 00684 mg, 2g of polyethylene glycol (400) and 1g of 95% ethanol are dissolved, and water for injection is added to dilute the solution to 100 mL; thus, a ZL0060.84mg/mL solution was obtained.

1.8 formulation B3: dissolving 168mg of (+) 2-anethol, 2g of polyethylene glycol (400) and 1g of 95% ethanol, and adding water for injection to dilute to 100 mL; to obtain a solution of (+) 2-daminol at 1.68 mg/mL.

1.9 formulation B4: dissolving 84mg of (+) 2-anethol, 2g of polyethylene glycol (400) and 1g of 95% ethanol, and adding water for injection to dilute to 100 mL; to obtain a solution of (+) 2-daminol 0.84 mg/mL.

1.10 formulation B5: ZL 00642 mg, 2g of polyethylene glycol (400) and 1g of 95% ethanol are dissolved, and water for injection is added to dilute the solution to 100 mL; thus obtaining a ZL0060.42mg/mL solution.

1.11 formulation B6: dissolving 42mg of (+) 2-anethol, 2g of polyethylene glycol (400) and 1g of 95% ethanol, and adding water for injection to dilute to 100 mL; to obtain a solution of (+) 2-daminol 0.42 mg/mL.

1.12 formulation B7: 21mg of (+) 2-anethol, 2g of polyethylene glycol (400) and 1g of 95% ethanol, and adding water for injection to dilute the mixture to 100 mL; to obtain a solution of (+) 2-daminol 0.21 mg/mL.

Example 2: protective effect of composition of ZL006 and (+) 2-anethol on focal cerebral ischemia reperfusion

2.1 preparation of focal cerebral ischemia reperfusion model

A Middle Cerebral Artery Occlusion (MCAO) cerebral ischemia reperfusion model is prepared by adopting an internal carotid artery embolization method. After animals were anesthetized with 7% chloral hydrate (6Ml/kg), fixed on an operating table in the prone position, the skin was sterilized, the neck was opened in the middle, the right common carotid artery, external carotid artery, internal carotid artery were separated, the vagus nerve was gently peeled off, the external carotid artery was ligated and cut off, the internal carotid artery was followed forward, and the pterygopalatine artery was ligated. Clamping the proximal end of the common carotid artery, making an incision from the distal end of a ligature of an external carotid artery, inserting a nylon wire with the outer diameter of 0.285mm, entering the internal carotid artery after the common carotid artery bifurcates, then slowly inserting until slight resistance exists (about 20mm from the bifurcation), blocking all blood supply of the middle cerebral artery, slightly pulling out the nylon wire after 2.0h of right cerebral ischemia, recovering the blood supply for reperfusion, simultaneously administering the drug through the tail vein, suturing the skin, and sterilizing.

2.2 cerebral protective Effect of a combination of ZL006 and (+) 2-anethol on acute focal cerebral ischemia in mice

After the animal is killed, cutting off the head and taking out the brain, removing the olfactory bulb, cerebellum and lower brainstem, flushing bloodstain on the surface of the brain with normal saline, sucking residual waterstain on the surface, placing at-20 ℃ for 20min, taking out, immediately making a coronal section vertically downwards on a sight line cross plane, cutting one section backwards at intervals of 2mm, placing the brain section in 1% TTC staining solution for incubation (37 ℃ for 90min), staining normal brain tissue into deep red, and ischemic brain tissue into pale, flushing with normal saline, quickly arranging the brain section in a row from front to back in sequence, sucking residual waterstain on the surface, and taking a picture.

And processing the picture by using Image J software, and calculating the corresponding area of the left brain and the area of the infarct focus according to a formula to calculate the percentage of the infarct focus.

Infarct volume calculation method:

V＝t(A1+A2+A3+………+An)

t is the slice thickness and A is the infarct size.

％I＝100％×(VC-VL)/VC

% I is the infarct volume percentage, VC is the control side (left hemisphere) brain volume, and VL is the infarct side (right hemisphere) non-infarct zone volume.

The effects of each group on the cerebral infarct size are shown in tables 1 and 2, and fig. 1 and 2.

TABLE 1 Effect of composition of ZL006 and (+) 2-Geraniol on cerebral infarct size in rats (Mean. + -. SEM)

Note: p <0.05, compared to model group

The above results show that: the composition of ZL006 and (+) 2-linalool can achieve significantly better anti-stroke efficacy at smaller doses.

TABLE 2 Effect of composition of ZL006 and (+) 2-anethol (3: 1) on cerebral infarct size in rats (Mean + -SEM)

Note: p <0.05, compared to model group

The above results show that: the composition of ZL006 and (+) 2-anethol has a definite dose-effect relationship and can obtain a remarkably better anti-stroke drug effect at a smaller dose.