CN116585447A - Cerebral apoplexy combined medicine and application thereof - Google Patents

Abstract

The invention discloses an application of emlicarbazen or pharmaceutically acceptable salts thereof in combination with an embedded alcohol compound or pharmaceutically acceptable salts thereof in preparing medicaments for preventing and/or treating cerebral apoplexy. The invention also discloses a combined medicament for preventing and/or treating cerebral apoplexy, wherein the active ingredients of the combined medicament comprise the emtriclosan or pharmaceutically acceptable salt thereof and the embedded alcohol compound or pharmaceutically acceptable salt thereof, the emtriclosan or pharmaceutically acceptable salt thereof and the embedded alcohol compound or pharmaceutically acceptable salt respectively become independent administration units, or the emtriclosan or pharmaceutically acceptable salt thereof and the embedded alcohol compound or pharmaceutically acceptable salt form a combined administration unit together. The pharmaceutical composition disclosed by the invention can be used for synergistically treating cerebral apoplexy, and the cerebral infarction range is obviously reduced.

Description

Cerebral apoplexy combined medicine and application thereof

Technical Field

The invention belongs to the field of biological medicine, and in particular relates to an enlicarbazepine combined medicine for treating cerebral apoplexy and application thereof.

Background

Cerebral stroke (stroke), also known as "stroke", "cerebrovascular accident" (cerebralvascular accident, CVA), is an acute cerebrovascular disease, a group of diseases that cause damage to brain tissue due to sudden rupture of cerebral vessels or failure of blood to flow into the brain due to vessel occlusion, including ischemic and hemorrhagic strokes. The cerebral apoplexy has the characteristics of high morbidity, high mortality and high disability rate. Cerebral stroke is the second most fatal disease worldwide, and its high recurrence rate and high disability rate leave about 75% of survivors with functional disability, severely jeopardizing patient health and quality of life. The cerebral apoplexy has the characteristics of high mortality rate and high disability rate.

At present, the treatment for acute ischemic cerebral apoplexy is mainly general treatment and specific treatment. Specific treatment is a key for treating ischemic cerebral apoplexy, and mainly comprises improving cerebral blood circulation, neuroprotection and the like. Intravenous thrombolysis treatment, intravascular treatment (thrombus taking) and the like are still the most effective treatment measures for acute ischemic cerebral apoplexy within a time window (the morbidity is less than 6 hours), but are difficult to meet the current national conditions. Typically ischemic stroke patients will take an average of 20.1 hours from onset to hospital, so the total proportion of patients who actually receive thrombolytic therapy is less than 3%.

Treatment of stroke is highly time dependent, with 1.9 billion neurons and 138 billion synapses lost every 1 minute of delay. The method is beneficial to timely diagnosis and treatment and rehabilitation nursing of cerebral apoplexy patients, so that the patients can be effectively treated, the death rate is reduced, and meanwhile, the problem that the related large-scale research of foreign neuroprotectants is less is considered, so that the recommendation of the neuroprotectants is relatively low. If the cerebral apoplexy patient is a cerebral apoplexy patient in vast rural areas, and if the medical level is insufficient, resident health consciousness is weak, road traffic is unsmooth, and the optimal treatment time is missed frequently, so that the death rate is high. Therefore, in the prevention and treatment of cerebral apoplexy, more choices must be considered to protect neurons of cerebral apoplexy patients, so as to reduce cerebral apoplexy death and disability loss and reduce social and economic burden. When the stroke occurs, the earlier the neuroprotection is performed, the progress of the death of the brain nerve is lightened, the damage of harmful substances to the brain can be effectively prevented, and more time is won for the brain.

Currently, edaravone and butylphthalide are main neuroprotective drugs clinically in China, and other drugs include citicoline, monosialotetrahexosyl ganglioside sodium, cinepazide and the like, but the overall treatment effect is not ideal, the cap of auxiliary drugs is difficult to extract, and development of novel effective and safe anti-cerebral apoplexy drugs or pharmaceutical compositions is needed to meet clinical urgent demands.

Since cerebral ischemic injury is associated with various mechanisms such as excessive formation of free radicals, toxic effects of excitatory amino acids, intracellular calcium overload, inflammatory response, etc., no matter what mechanism is, the final outcome leads to neuronal cell death, functional destruction, and formation of cerebral infarction foci. The cell death mode mainly comprises apoptosis and necrosis. Nerve cell apoptosis and necrosis coexist in cerebral ischemia injury. The ischemic core is dominated by cellular necrosis, while the surrounding bright and dark bands are dominated by apoptosis.

Apoptosis mainly includes both endogenous apoptotic pathways (mitochondrial-mediated apoptotic pathways) and exogenous apoptotic pathways (death receptor-mediated apoptotic pathways). The Caspase family is a major class of regulatory factors in apoptosis, the initiator and the final executor of apoptosis. In cerebral ischemia, death receptor mediated apoptosis pathway activates caspase-8, further activates its downstream effector protease caspase-3 and activates mitochondrial apoptosis pathway leading to apoptosis. Research shows that inhibiting caspase activity can prevent the development of nerve apoptosis, reduce cerebral ischemia damage and reduce infarct range. The caspase inhibitor, emtricosan, can specifically inhibit caspase-8 and caspase-3/7, thus preventing the progress of apoptosis caused by cerebral ischemia and cerebral apoplexy, reducing the degree of cerebral ischemia injury and reducing the infarct size.

The blood brain barrier is a diverse and complex dynamic system, which allows the passage of nutrients required by brain tissues in the blood circulation, effectively blocks the invasion of harmful substances, discharges metabolites, and reacts according to various needs of the components in the brain, thereby maintaining the brain environment constant and ensuring the normal structure and function of the central nervous system. However, the blood brain barrier also prevents the medicine from entering the brain of a human body, so that the medicine is difficult to penetrate through the blood brain barrier to reach the effective treatment concentration in the brain, thereby causing difficulty in treating brain diseases such as cerebral apoplexy, brain tumor, central nervous system infection, medicine addiction and the like.

The embedded alcohol is also called borneol, borneol and the like, is a common medicament and has wide pharmacological actions, such as: analgesic, anti-inflammatory, antibacterial, antithrombotic, etc., most prominent is the effect of promoting the permeation of drugs through the blood brain barrier. In long-term clinical application, the weakness of the embedded alcohol is gradually revealed, such as the characteristics of weak drug effect, large irritation, strong volatility, existence of certain toxic and side effects and the like, which become main factors influencing the clinical application of the embedded alcohol. A large number of researches show that the absorption speed of the embedded alcohol in the body is high, the embedded alcohol can easily pass through the blood brain barrier, the physiological stability of the brain can be ensured, the blood brain barrier can be opened, the pathological edema can be greatly affected, the concentration of other medicines in the brain can be improved, and therefore, the embedded alcohol can protect the nervous system.

Cerebral edema can be caused during cerebral ischemia, and the blood brain barrier is further aggravated, so that the therapeutic medicine cannot enter the brain rapidly and sufficiently to play a role.

Enlicarson (Emricasan, 1) is a potent, oral caspase inhibitor with the chemical name N- [2- (tert-butyl) phenyl ] -2-oxoglycyl-N- [ (1S) -1- (carboxymethyl) -2-oxo-3- (2, 3,5, 6-tetrafluorophenoxy) propyl ] -L-alaninamide. Enlicarson aims to reduce the activity of enzymes that mediate inflammation and cell death or apoptosis, by reducing the activity of which Enlicarson may block the progression of the entire liver disease. Conatus pharmaceutical company announced that phase ii clinical trials of emtricosan (emricasan) for the treatment of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis achieved successful results.

At present, no report on the combined application of the emlicarbazen and the embedded alcohol exists in the prior art, and particularly no combined application of the emlicarbazen and the embedded alcohol in the treatment of ischemic cerebral apoplexy exists.

The invention provides a combined drug preparation of emlicarbazepine and embedded alcohol, wherein the emlicarbazepine can inhibit nerve cell apoptosis by inhibiting caspase enzyme activity, the embedded alcohol can open blood brain barrier and improve pathological edema, and meanwhile, the concentration of the emlicarbazepine in the brain is increased, and the synergic effect greatly reduces cerebral ischemic injury.

Disclosure of Invention

The invention aims to provide a combined drug of emlicarbazen and embedded alcohol, which is used for treating cerebral apoplexy.

In order to achieve the above object, the present invention includes the following technical solutions.

In a first aspect, the present invention provides an use of emlicarbazen or a pharmaceutically acceptable salt thereof in combination with an alcohol compound or a pharmaceutically acceptable salt thereof for preparing a medicament for preventing and/or treating cerebral apoplexy.

In a second aspect of the present invention, there is provided a pharmaceutical combination for preventing and/or treating cerebral apoplexy, wherein the active ingredients include emlicarbazepine or a pharmaceutically acceptable salt thereof, and an alcohol-embedded compound or a pharmaceutically acceptable salt thereof, wherein the emlicarbazepine or the pharmaceutically acceptable salt thereof and the alcohol-embedded compound or the pharmaceutically acceptable salt thereof are respectively formed into separate administration units, or the emlicarbazepine or the pharmaceutically acceptable salt thereof and the alcohol-embedded compound or the pharmaceutically acceptable salt thereof together form a combined administration unit.

In some embodiments, the active ingredient of the combination is emtriclosan or a pharmaceutically acceptable salt thereof, and the glycol compound or pharmaceutically acceptable salt is a pharmaceutically acceptable salt, such as hydrochloride, sulfate, phosphate, acetate, benzoate, fumarate, maleate, citrate, tartrate, mesylate, ethanesulfonate, benzenesulfonate, crescent sulfonate, hydroquinone sulfonate and p-toluene sulfonate.

The active ingredients of the pharmaceutical composition for treating cerebral apoplexy consist of the emlicarbazen and the embedded alcohol according to the weight ratio of 1 (0.05-0.25), and the two ingredients exist in a free state or in a pharmaceutically acceptable salt form or in optical isomers, derivatives and the like. The emlicarbazepine is suitable for the embedded alcohol in the form of pharmaceutically acceptable salt and optical isomer and derivative, wherein the optical isomer can be dextro-isomer, levo-isomer and the mixture of the dextro-isomer and the levo-isomer.

In some embodiments, the active ingredients of the pharmaceutical composition comprise (by weight ratio) 1 (0.05-0.15) of emtriclosan and embedded alcohol, and further comprise (by weight ratio) 1:0.08-0.1.

More preferably, the active ingredients of the pharmaceutical composition are composed of the emlicarbazen and the embedded alcohol according to the weight ratio of 1:0.1.

The dosage form of the combined drug is not limited, and any pharmaceutically acceptable dosage form can be prepared according to the known technology, so long as the dosage form can enable the active ingredients to effectively reach the in vivo, and the combined drug comprises the following components: injection, tablet, film coated tablet, enteric coated tablet, capsule, buccal preparation, granule, powder, unguent, pellet, suspension, pill, powder, spray, drop, suppository, cream, patch, etc.; injection forms are preferred, such as: liquid injection, powder injection, concentrated solution for injection, etc.; injection forms include, but are not limited to, crude drug forms, drug-loaded nanoparticles, and the like; injection sites include, but are not limited to, intradermal injection, subcutaneous injection, intramuscular injection, intravenous injection, spinal cavity injection, and the like. Intravenous administration is preferred.

In some embodiments, the stroke comprises ischemic stroke, hemorrhagic stroke, acute stroke, and stroke with mild stroke. Preferably, the cerebral apoplexy refers to acute ischemic cerebral apoplexy.

Compared with the prior art, the invention has the beneficial effects that:

(1) The synergistic treatment of cerebral apoplexy, in particular to the reduction of cerebral infarction scope, the improvement of long-term survival rate and the reduction of disability rate of patients, and can be suitable for acute ischemic cerebral apoplexy;

(2) Adverse reactions of the medicine are reduced;

(3) The concentration of the medicine in the brain is improved;

animal experiments prove that the emlicarbazen, the embedded alcohol and the composition of the emlicarbazen and the embedded alcohol have the effect of treating cerebral apoplexy, wherein the curative effect of the composition of the emlicarbazen and the embedded alcohol is optimal.

In conclusion, in the pharmaceutical composition, the emlicarbazepine can inhibit nerve cell apoptosis by inhibiting caspase enzyme activity, the embedded alcohol can open blood brain barrier, has an improvement effect on pathological edema, and simultaneously improves the concentration of the emlicarbazepine in the brain, so that the synergic effect greatly reduces the cerebral infarction range caused by cerebral ischemic injury.

Drawings

FIG. 1 shows a TCC staining pattern for each group of cerebral infarction in example 1.

Detailed Description

The present invention will be described more fully hereinafter in order to facilitate an understanding of the present invention. This invention may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

The experimental procedures, which do not address the specific conditions in the examples below, are generally carried out under conventional conditions or under conditions recommended by the manufacturer. The various chemicals commonly used in the examples are commercially available.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.

Definitions to facilitate understanding of the present technology, some terms and phrases are defined below.

Throughout the specification and claims, the following terms have the meanings explicitly associated herein, unless the context clearly dictates otherwise. The phrase "in one embodiment" as used in the present invention does not necessarily refer to the same embodiment, although it may. Furthermore, the phrase "in another embodiment" as used in the present invention does not necessarily refer to a different embodiment, although it may. Accordingly, as described below, various embodiments of the present invention may be readily combined without departing from the scope or spirit of the present invention.

In the present invention, the term "plurality" means two or more. "and/or", describes an association relationship of an association object, and indicates that there may be three relationships, for example, a and/or B, and may indicate: a exists alone, A and B exist together, and B exists alone. The character "/" generally indicates that the context-dependent object is an "or" relationship. The present invention will be described in further detail with reference to specific examples.

Hereinafter, the amounts of emlicarbazide and the right-embedding alcohol are expressed as equivalent amounts of emlicarbazide and the right-embedding alcohol in the free form without salification.

Animal experiment: protection of ischemic stroke by emtriclosan and dexanabinol compositions

And (3) implementing the medicine:

enlicarpa is purchased from Haoyuan biological medicine technologies Co., ltd.

Right-hand methanol was purchased from Wohan Green Jing Fenghua Biotechnology Co.

Edaravone right-hand-embedding injection (specification: 5ml (10 mg:2.5 mg)) was purchased from the Md.

The composition of emlicarbazen and dextrol was prepared into an injectable drug solution using 100mM phosphate buffer (pH 7.4) as a vehicle.

Example 1

Experimental animals:

50 male SD rats, the weight of which is 250-300g, are kept in a barrier system, the environmental conditions of an animal house are controlled to be between 20 and 26 ℃ and the humidity of which is 40-70 percent, the environment is recorded by a raiser every day, every afternoon, 10 to 20 times are replaced every hour in the room, the lighting in the animal house is alternated in 12/12 hours of light and shade circulation (except for the interruption necessary in the experimental activity), and any interruption is recorded. Animals need to be kept in the closed cages, and after the animals enter the group experiment, the animals are kept in the closed cages. The animal cage is made of polysulfone plastic and is provided with corncob padding, and toys for biting or shielding are arranged in the cage to serve as an environmental enrichment measure for each cage animal. Each cage is provided with a cage card for identifying the group.

The modeling method comprises the following steps:

animal anesthesia depth was maintained with 2.5% isoflurane respiratory anesthesia and a cerebral ischemia reperfusion model was established as follows:

(1) shaving the neck hair of the animal;

(2) sterilizing the operation area;

(3) cutting the epidermis of the neck to expose the left common carotid artery;

(4) ligating the common carotid artery, the pterygopalatine artery and the external carotid artery at the proximal end;

(5) cutting a small opening at the common carotid artery which is 2mm away from the bifurcation of the internal and external carotid arteries;

(6) nylon wire (3-0) is used for being inserted from the small opening of the common carotid artery (the depth is 18 mm-20 mm) and enters the internal carotid artery to reach the left middle cerebral artery;

(7) fixing the wire bolt;

(8) pulling out the wire plug after 120min, and opening the external carotid artery to realize ischemia reperfusion;

model success criteria:

1: scoring according to the Longa method within 30-80 min after operation, and grouping rats with scores of 2-3, wherein animals with scores of 4 or less than 1 are removed.

Grouping and administration:

after the molding, the test was divided into 5 groups according to animal weights, and 10 animals in each group were respectively:

group 1-model Group (n=10), vehicle was immediately administered when the plug was withdrawn and refilled, and the injection volume was 5mL/kg, and single administration was performed for 1 day;

group 2-edaravone right embedded alcohol injection positive medicine Group (N=10), edaravone 12 mg/kg+right embedded alcohol 3mg/kg, immediately administration when the plug is pulled out for recharging, the injection volume is 5mL/kg, single administration and observation is carried out for 1 day;

group 3-composition Group (n=10), emlicarbazen (5 mg/kg) +right-hand methanol (0.5 mg/kg), was administered immediately upon extacting the plug and re-filling, with injection volume of 5mL/kg, single administration, and observation for 1 day;

group 4-emlicarbazepine single Group (n=10), 5mg/kg, immediately administration upon extacting of the plug and recharge, single administration at 5mL/kg injection volume, observation for 1 day;

group 5-right monol (n=10), 0.5mg/kg, immediately administered upon extacting of the plug and recharge, injection volume at 5mL/kg, single administration, observation for 1 day;

cerebral infarction evaluation:

animals were euthanized 1 day after cerebral ischemia reperfusion (24 hours of reperfusion), brain tissue was harvested, coronally cut, 3mm thick per slice, 2% TTC stained, infarcted areas were white after staining, and non-infarcted areas were red. After double-sided scanning by using a Canon scanner, calculating cerebral infarction conditions according to the average area of the front side and the back side, wherein the calculation formula is as follows: infarct percentage= [1- (left brain area-left brain infarct area)/right brain area ]. 100%. Brain tissue was then preserved using 10% neutral formalin fixation. The data of animals successfully established by all models will be included in statistics, and the standard of success of the experimental model is that the cerebral infarction percentage is between 20% and 60%, animals with the infarction percentage less than 20% are considered to be unmolded, and animals with the infarction percentage greater than 60% are considered to be reperfusion failure, so that the infarct area is overlarge.

Experimental results:

see in particular table 1, fig. 1.

The model group has obvious white infarct focus, and the rat cerebral infarct focus of each administration group shows different amplitude reduction, which shows that each administration group has clinical improvement potential. The composition of the emlicarbazel and the right embedded alcohol has the best effect on the reduction range of cerebral infarction of rats, has extremely significant difference (P < 0.01) compared with a model group, has significant difference (P < 0.05) compared with the emlicarbazel group and the right embedded alcohol group, and shows that the pharmaceutical composition formed by combining the emlicarbazel and the right embedded alcohol has obvious treatment and improvement effects on ischemic cerebral apoplexy and is obviously superior to the single use of the emlicarbazel and the right embedded alcohol.

Table 1 percentage of cerebral infarct size for each group of animals

Group (N=8)	Percentage of cerebral infarct area (%)
		Model group, single intravenous administration vehicle	49.51±5.86
Edaravone right-embedded alcohol group, 12mg/kg, single intravenous administration	39.38±12.05*
		Enlicarson 5 mg/kg+Right-hand methanol 0.5mg/kg, single intravenous administration	21.18±9.32** #
Enlicarson, 5mg/kg, single intravenous administration	35.16±13.02*
		Right-hand alcohol, 0.5mg/kg, single intravenous administration	46.82±12.13

One-way ANOVA:**p<0.01vs. model group; * P is p<0.05vs. model group; ^# p<0.05vs. Enlicarson group, right embedded alcohol group.

In fig. 1, each group corresponds to the following:

1M001: a model group;

2M009: a positive group of edaravone right-embedding alcohol injection;

3M001: a group of 5mg/kg of Enlicarson plus 0.5mg/kg of Right-hand methanol;

4M002: enlicarson 5mg/kg group;

5M008: right embedded alcohol 0.5mg/kg group.

The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.

The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.

Claims (11)

1. The use of emtricin or pharmaceutically acceptable salts thereof in combination with an alcohol compound or pharmaceutically acceptable salt thereof for the preparation of a medicament for the prophylaxis and/or treatment of cerebral stroke.

2. The use according to claim 1, wherein the pharmaceutically acceptable salt is a hydrochloride, sulfate, phosphate, acetate, benzoate, fumarate, maleate, citrate, tartrate, mesylate, esylate, besylate, selenylate, hydroquinonesulfonate or p-toluenesulfonic acid.

3. The use according to claim 1, wherein the glycol compound is dextro-glycol.

4. The use according to claim 3, wherein the ratio of the amount of the emlicarbazepine to the amount of the dextro-embedded alcohol is 1:0.05-0.25, preferably 1:0.05-0.1.

5. The combined medicine for preventing and/or treating cerebral apoplexy is characterized in that the active ingredients comprise the emlicarbazide or pharmaceutically acceptable salt thereof and the embedded alcohol compound or pharmaceutically acceptable salt thereof, wherein the emlicarbazide or pharmaceutically acceptable salt thereof and the embedded alcohol compound or pharmaceutically acceptable salt respectively become independent administration units, or the emlicarbazide or pharmaceutically acceptable salt thereof and the embedded alcohol compound or pharmaceutically acceptable salt form a combined administration unit together.

6. The combination of claim 5, wherein the pharmaceutically acceptable salt is a hydrochloride, sulfate, phosphate, acetate, benzoate, fumarate, maleate, citrate, tartrate, mesylate, ethanesulfonate, benzenesulfonate, laurylsulfonate, hydroquinone sulfonate, or p-toluenesulfonic acid.

7. The combination according to claim 5, wherein the glycol compound is dextro-glycol.

8. The combination according to claim 7, wherein the ratio of the amount of the emtriclosan to the amount of the dextro-embedded alcohol is 1:0.05-0.25 by mass.

9. The combination according to claim 8, wherein the ratio of the amount of the emtriclosan to the amount of the dextro-embedded alcohol is 1:0.05-0.1.

10. The combination according to claim 5, wherein the cerebral apoplexy is ischemic cerebral apoplexy or hemorrhagic cerebral apoplexy.

11. The combination of claim 5, wherein the combination is an emlicarbazen and a dextro-embedded alcohol together forming a combined dosage unit, and wherein the dosage unit is in the form of an injection.