CN109939102B - Pharmaceutical composition containing butylphthalide and borneol and application thereof - Google Patents
Pharmaceutical composition containing butylphthalide and borneol and application thereof Download PDFInfo
- Publication number
- CN109939102B CN109939102B CN201910315555.8A CN201910315555A CN109939102B CN 109939102 B CN109939102 B CN 109939102B CN 201910315555 A CN201910315555 A CN 201910315555A CN 109939102 B CN109939102 B CN 109939102B
- Authority
- CN
- China
- Prior art keywords
- borneol
- butylphthalide
- pharmaceutical composition
- nbp
- oleum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a pharmaceutical composition, which comprises butylphthalide and borneol. The pharmaceutical composition provided by the invention has a synergistic effect in treating cerebrovascular diseases, especially ischemic cerebrovascular diseases, can remarkably increase the treatment effect of butylphthalide, reduce the effective dosage of butylphthalide, reduce the dosage of butylphthalide, and reduce the occurrence of adverse reactions after long-term application.
Description
The application is a divisional application of a patent application with the application date of 2013, 6 and 13, the application number of 201310231054.4 and the invention name of 'a pharmaceutical composition containing butylphthalide and borneol and application thereof'.
Technical Field
The invention relates to a pharmaceutical composition containing butylphthalide and borneol and application thereof in preparing a medicament for treating cerebrovascular diseases.
Background
Cerebrovascular disease (CVD) is a brain disorder caused by cerebrovascular abnormality, and stroke (stroke) generally refers to an acute cerebrovascular disease. According to survey data of the world health organization on 17 countries and regions, the incidence rate fluctuation of the cerebrovascular disease is 15-287/10 ten thousands, and the regional distribution is different. The average incidence rate of cerebrovascular diseases in China is about 130/110 ten thousands, which is higher than Europe and America and is close to Japan. The data research of all countries in the world shows that: cerebrovascular incidence increases with age. At present, China is facing the challenge of rapid aging, cerebrovascular diseases are the second leading cause seriously threatening the health and life of middle-aged and old people, and once cerebral apoplexy occurs, the disability rate and the death rate are high, over 75 percent of survivors have sequelae with different degrees, and the life quality of patients is seriously influenced. The future prospect of the Chinese cerebrovascular disease is extremely severe, and the Chinese cerebrovascular disease will form a huge burden of medical expenditure. Thus, cerebrovascular diseases have become a hot spot of medical research.
Cerebrovascular diseases can be simply classified into ischemic cerebrovascular diseases due to reduced blood flow or interruption of blood flow, and hemorrhagic cerebrovascular diseases due to rupture of blood vessels. Ischemic cerebrovascular disease (ICVD) refers to the thickening of the wall of a cerebral vessel, the stenosis of a vessel lumen and the formation of thrombus due to various reasons, so that the blood supply area of the affected cerebral vessel is subjected to ischemia and hypoxia, hypoxic necrosis and brain softening of lesion tissues are caused, and corresponding neurological dysfunction occurs. Ischemic cerebrovascular diseases are mainly cerebral infarction (including cerebral thrombosis and cerebral embolism); in addition to cerebral infarction, there is ischemic cerebrovascular disease which can be completely recovered within 24 hours without any sequelae, called transient ischemic attack or transient ischemic attack, and doctor's habit is abbreviated as TIA, also called minor stroke. Hemorrhagic cerebrovascular diseases are also divided into two categories, one is vascular rupture, and blood flows into the parenchyma of brain to become cerebral hemorrhage or cerebral hemorrhage; the other is vascular rupture and blood flows into the subarachnoid space surrounding the brain, which becomes subarachnoid hemorrhage, SAH for short.
Ischemic cerebrovascular disease causes impairment of brain function and impairment of nerve function in the blood supply region due to blood supply disturbance of the blood supply arteries of the brain. Brain tissue has a large demand for energy and oxygen, but has no reserve function. Therefore, once ischemic hypoxia occurs, metabolic disorders occur quickly, causing cell damage. Cerebral tissue blood supply failure in the blood supply area over a certain period of time can lead to cerebral infarction. It is considered that an ischemic penumbra (ischemic penumbra) is generally formed around the cerebral infarction focus, and a large number of intact brain cells in a dormant state or a semi-dormant state, which can only maintain the morphology of the brain cells, exist. Because cerebral blood flow is blocked, the device lacks energy supply and cannot run the original normal function. The presence of a penumbra is generally around 6-24 hours. In this period, if the blood supply disorder is timely relieved and the blood flow in the ischemic area is recovered, the necrosis of the ischemic tissue can be avoided and the diseased tissue can be saved. If the degree of ischemia is further increased, the integrity of the cell membrane is impaired, and nerve cells are necrosed, and this stage becomes irreversible damage. In recent years, with the clinical popularization and application of interventional techniques, thrombolysis and other methods, even though local stenosis is relieved and tissues and organs with ischemic lesions are supplied with blood again, it has been found in clinical and animal experiments that even after a certain period of time, measures are taken to restore blood supply to tissues and organs with blood supply disorders, tissue damage and organ dysfunction are not relieved, and the phenomenon that clinical symptoms are rather aggravated is called cerebral ischemia-reperfusion injury (CIRI), and even serious patients cause cerebral edema and hemorrhage. The pathogenesis of cerebral ischemia-reperfusion injury is very complex, and various factors are mutually interwoven, cause and effect and mutually influence. Previous studies have shown that: free radical damage, hypoxia and energy metabolism failure, inflammatory injury, calcium ion overload, excitatory amino acid toxic action and the like are related and jointly involved, and finally nerve cell necrosis and functional symptom loss are caused.
Therefore, the treatment of cerebrovascular diseases, especially ischemic cerebrovascular diseases, is a great problem in the current clinical treatment.
Butylphthalide (3-n-butylphthalide), also known as apigenin A, is an effective component separated from celery seeds, and is clinically applied to racemic butylphthalide and levo-butylphthalide, and the structural formulas are respectively as follows:
butylphthalide is a multi-target ischemic cerebrovascular disease drug, and acts on a plurality of pathological links caused by cerebral ischemia through mechanisms of improving the levels of cerebrovascular endothelial NO (nitric oxide) and PGI2 (prostacyclin), reducing the intracellular calcium concentration, inhibiting the release of glutamic acid, reducing the content of arachidonic acid, inhibiting oxygen free radicals, improving the activity of antioxidant enzyme and the like. Animal pharmacodynamic studies show that butylphthalide has strong anti-cerebral ischemia effect; obviously improve microcirculation and blood flow volume of ischemic brain area, increase the number of capillary vessels of ischemic area; relieving cerebral edema and reducing the infarct area of local cerebral ischemia of rats; improving brain energy metabolism, and inhibiting nerve cell apoptosis; inhibiting thrombosis, etc. Clinical research results show that butylphthalide has obvious effect of improving ischemic nerve function damage and can promote the function recovery of patients. When the butylphthalide medicine is clinically applied, gastrointestinal adverse reactions such as slight increase of transaminase, nausea, abdominal pain and the like are occasionally caused.
At present, the butylphthalide drugs used in clinical application include butylphthalide soft capsules and butylphthalide sodium chloride injection. According to the specification, the specification of the butylphthalide soft capsule is as follows: 0.1g, the usage and dosage are as follows: orally administered on an empty stomach, two granules (0.2g) at a time, four times a day, and ten to twelve days as a treatment course; the specification of the butylphthalide sodium chloride injection is as follows: 100ml, 25mg of butylphthalide and 0.9g of sodium chloride, and the usage and dosage are as follows: intravenous drip is administered 2 times daily, each time 25mg (100ml), with the interval of two times not less than 6 hr, and the treatment course is 14 days. Most patients with ischemic cerebrovascular diseases have inconvenient movement and diet and long treatment period, so in order to improve the medication compliance of the patients, the treatment effect of butylphthalide needs to be further improved, such as reduction of the effective dose and reduction of the dosage.
Borneol (also known as borneol) is a commonly used traditional Chinese medicine. Natural borneol, borneol and synthetic borneol are included in the first part of pharmacopoeia 2010 edition of the people's republic of China. Natural borneol (also called plum slice) is processed product of Dryobalanops aromatica Gaertn.f resin (called plum slice) of Dryobalanops aromatica of Dipterocarpaceae, and contains D-borneol ((+) -borneo1) as main ingredient, with D-borneol content not less than 96%; the Blumea balsamifera (L.) DC tablet is crystal of Blumea balsamifera (L.) DC leaf extract of Compositae, mainly contains levo-borneol ((-) -borneo1), and the levo-borneol content is not less than 85% of borneol; the synthetic borneol is a refined product (also called machine-made borneol) synthesized by a chemical method by using turpentine or camphor as a raw material, is a racemate, and contains a certain isoborneo1, wherein the borneol content is not less than 55%.
In addition, d-borneol is also called (+) -2-borneol, and l-borneol is also called (-) -2-borneol.
Borneol is pungent and bitter in taste and slightly cold in nature, and has the functions of inducing resuscitation, refreshing mind, clearing heat and relieving pain, and can be used for treating fever unconsciousness, convulsion syncope, apoplexy, phlegm syncope, pharyngitis, toothache, aphtha, carbuncle and sore, conjunctival congestion, etc. Borneol has the effects of resuscitation, aromatic curtain and guiding the medicine upwards, and is often used as a guiding medicine to improve the treatment effect of other medicines; ben Cao Yan Yi (the Yan Yi of materia Medica) points out that borneol is weak in one way and active in adjuvant therapy.
The borneol has the best effect by using natural borneol, but because of resource limitation, synthetic borneol is mostly used in the existing Chinese patent medicines to replace the natural borneol. In addition, because the content of the d-borneol in the natural borneol is as low as 96 percent and still contains a certain amount of impurities, which may affect the safety of medication, people purify the natural borneol, and the (+) -2-borneol with the purity of more than or equal to 98 percent is commercially available at present.
The daily dose of borneol specified in the pharmacopoeia is 150-300mg per day, and the large dose of borneol can cause adverse gastrointestinal reactions such as heartburn, nausea, abdominal pain, vomiting and the like.
Disclosure of Invention
The invention aims to provide a pharmaceutical composition, which comprises butylphthalide and borneol, and has a synergistic effect when used cooperatively, so that the effect of treating cerebrovascular diseases, especially ischemic cerebrovascular diseases, can be improved.
Wherein:
the butylphthalide comprises levo-butylphthalide, a mixture of levo-butylphthalide and dextro-butylphthalide, especially racemic butylphthalide, preferably racemic butylphthalide and levo-butylphthalide, more preferably levo-butylphthalide.
The borneol is borneol containing borneol with any content, preferably natural borneol, synthetic borneol, (+) -2-borneol with the purity of more than or equal to 98 percent and (-) -2-borneol, and more preferably natural borneol and (+) -2-borneol with the purity of more than or equal to 98 percent.
In the pharmaceutical composition, butylphthalide and borneol may be present in any suitable ratio. The weight ratio of butylphthalide to borneol is preferably 100: 1-1: 100, more preferably 2.5-80: 1, further preferably 5-40: 1, and further preferably 10-40: 1, from the comprehensive consideration of the treatment effectiveness, safety, medication cost and the like.
The pharmaceutical composition can also contain pharmaceutically acceptable auxiliary materials. The auxiliary materials are related to the prepared dosage form.
Because the butylphthalide and the borneol are both fat-soluble substances, and the butylphthalide soft capsule and the injection are products on the market, the preparation of the butylphthalide soft capsule has a mature process, for example, CN1623542A discloses a preparation process of the butylphthalide soft capsule, and CN1394880A discloses a preparation process of the butylphthalide injection. Therefore, the pharmaceutical composition is preferably formulated into soft capsules or injections. If the soft capsule is prepared, the auxiliary material is vegetable oil, and an antioxidant is optionally added; the vegetable oil can be one of oleum Sesami, oleum Maydis, oleum Arachidis Hypogaeae, soybean oil, oleum Armeniacae amarum, oleum Persicae, oleum gossypii semen, oleum Helianthi, and oleum Olivarum, preferably soybean oil; the antioxidant is preferably dibutyl carboxytoluene. If the injection is prepared, a solvent and a cosolvent can be added, wherein the solvent is preferably water for injection and ethanol, and the cosolvent is preferably cyclodextrin or cyclodextrin derivative, and more preferably hydroxypropyl-beta-cyclodextrin.
The invention also aims to provide an application of the pharmaceutical composition in preparing a medicament for treating cerebrovascular diseases.
Preferably, the pharmaceutical composition is applied to the preparation of medicines for treating ischemic cerebrovascular diseases or cerebral infarction.
The pharmaceutical composition provided by the invention comprises butylphthalide and borneol, has a synergistic effect in treating cerebrovascular diseases, particularly ischemic cerebrovascular diseases, can remarkably increase the treatment effect of butylphthalide, reduce the effective dose of butylphthalide, reduce the dosage of the butylphthalide, and reduce the occurrence of adverse reactions of the butylphthalide after long-term application.
Detailed Description
The present invention is further illustrated by the following examples, which are not intended to limit the scope of the present invention in any way. In the embodiment, the natural borneol refers to natural borneol collected in pharmacopoeia of the people's republic of China in 2005 edition, namely d-borneol.
Example 1: preparation of pharmaceutical solutions for injection
(1) Preparation of 30% HP-beta-CD
Taking 600g of HP-beta-CD (hydroxypropyl-beta-cyclodextrin), adding 1200mL of injection water, stirring for dissolving, and finally fixing the volume to 2000mL by using the injection water, namely the 30% HP-beta-CD.
(2) Preparation of butylphthalide solution for injection
Respectively weighing 4g of racemic butylphthalide and 4g of levo-butylphthalide, respectively adding 600mL of 30% HP-beta-CD, and stirring until the solution is clear.
(3) Preparation of borneol solution for injection
Respectively adding 1g of natural borneol into 20mL of 95% ethanol, uniformly mixing, adding 280mL of 30% HP-beta-CD, and stirring until the solution is clear.
(4) Preparation of butylphthalide and borneol solution for injection
Mixing butylphthalide solution and Borneolum solution at given ratio, and mixing at the following ratio:
the above solution can also be prepared into injection by conventional method such as aseptic processing and subpackaging.
Example 2: preparation of oral medicinal liquid oil
(1) Preparation of butylphthalide medicinal liquid oil
Respectively weighing 25g of racemic butylphthalide and 25g of levo-butylphthalide, respectively adding into 225g of soybean oil, and sufficiently and uniformly stirring to obtain the composition.
(2) Preparation of borneol medicinal liquid oil
Adding 10g of natural borneol into 90g of soybean oil, and stirring and dissolving until the solution is clear.
(3) Preparation of butylphthalide + borneol tablet liquid medicine oil
Mixing butyl phthalide liquid medicine oil and borneol liquid medicine oil in given proportion uniformly to obtain the product, wherein the mixing proportion is as follows:
| butylphthalide medicinal liquid oil | Borneol medicinal liquid oil | The weight ratio of butylphthalide to borneol |
| 20g | 2g | 10:1 |
| 40g | 2g | 20:1 |
| 80g | 2g | 40:1 |
The above medicinal liquid oil can also be made into soft capsule by conventional method such as soft capsule preparation method disclosed in CN 1623542A.
Example 3: mouse ischemia test (borneol dosage investigation)
1 animal experiment Material
1.1 drug test drugs: example 1 prepared racemic butylphthalide (NBP) solution for injection, natural borneol solution for injection, racemic butylphthalide + natural borneol solution for injection, model control was 30% HP-beta-CD.
1.2 animals SPF-grade KM mice, male, 18-22 g, provided by the experimental animals center of north river medical university; the production license number of the experimental animal is SCXK (Ji) 2008-1-003. License number for experimental animals: SYXK (Ji) 2011-.
1.3 data processing data was analyzed for variance and inter-group testing by SPSS11.5 statistical software.
2 mouse Whole brain ischemia test
2.1 animal groups and administrations: 100 Kunming mice are randomly divided into 10 groups, namely a model control group (30% HP-beta-CD) and an NBP group (10mg kg)-1) Natural ice sheets (0.25, 0.5, 1 and 2 mg/kg)-1) NBP + Natural Ice sheet group (NBP 10 mg/kg)-10.25, 0.5, 1 and 2mg of natural borneolkg-1) 10 per group.
2.2 test operation and index observation: 30min after intravenous administration of the mice, the head of the animals is broken at the back of ears and ears, and the head breaking and breathing time and the breathing frequency of the mice are observed.
3 results of the test
The electrical activity of the brain depends on continuous energy metabolism, the ischemia and anoxia after the mouse breaks the head cause the brain death, and the anti-cerebral ischemia effect of the medicine is investigated through the improvement effect of the medicine on the tolerance degree of the ischemia and anoxia. As shown in Table 1, NBP was administered in a single intravenous dose of 10 mg/kg-1Can obviously prolong the breathing times and breathing time of the mouse after the head is broken, and has obvious difference compared with a model group. Borneol 0.25, 0.5, 1.0 and 2.0 mg/kg-1Intravenous administration alone had no effect on whole brain ischemia in mice. 0.25, 0.5, 1 and 2mg kg-1The borneol and NBP combined administration has the tendency of prolonging the breathing times and breathing time of mice after the mice are broken, and the combined administration of the borneol and the NBP is suggested to enhance the protective effect of the NBP on the mice with ischemia and anoxia, but the treatment effect is not in direct proportion to the borneol administration amount, and the borneol administration amount is 0.5mg kg from the comprehensive consideration of the treatment effect, the safety and the administration cost-1Most preferably, 1 mg/kg-1Next, the method is described. Therefore, the following examples select the borneol dosage of 0.5mg/kg-1Synergy was investigated in combination with NBP.
TABLE 1 Effect on Whole brain ischemia in mice: (
n＝10)
P <0.05, p <0.01, compared to model groups.
Therefore, the following examples select a borneol dose of 0.5mg/kg-1Synergy was investigated in combination with NBP.
Example 4: mouse global ischemia test
1 animal experiment Material
1.1 drug test drugs: example 1 prepared racemic butylphthalide (NBP) solution for injection, natural borneol solution for injection, racemic butylphthalide + natural borneol solution for injection, model control was 30% HP-beta-CD.
1.2 animals SPF-grade KM mice, male, 18-22 g, provided by the experimental animals center of north river medical university; the production license number of the experimental animal is SCXK (Ji) 2008-1-003. License number for experimental animals: SYXK (Ji) 2011-.
1.3 data processing data was analyzed for variance and inter-group testing by SPSS11.5 statistical software.
2 mouse Whole brain ischemia test
2.1 animal groups and administrations: 80 Kunming mice were randomly divided into 8 groups, namely model control group (30% HP-beta-CD) and NBP group (20, 10 and 5 mg.kg)-1) NBP + Natural Ice sheet group (NBP 20, 10 and 5 mg. kg)-10.5mg/kg of natural borneol-1) And natural ice tablet group (0.5 mg/kg)-1) 10 per group.
2.2 test operation and index observation: 30min after intravenous administration of the mice, the head of the animals is broken at the back of ears and ears, and the head breaking and breathing time and the breathing frequency of the mice are observed.
3 results of the test
The electrical activity of the brain depends on continuous energy metabolism, the ischemia and anoxia after the mouse breaks the head cause the brain death, and the anti-cerebral ischemia effect of the medicine is investigated through the improvement effect of the medicine on the tolerance degree of the ischemia and anoxia. NBP was administered in a single intravenous dose of 20 and 10 mg-kg as shown in Table 2-1All can prolong the gasping times of the broken mouse, 20 mg/kg-1Can prolong the gasp time of the mouse after the head is broken, and has significant difference compared with a model group. 0.5mg/kg of borneol-1Intravenous administration alone had no effect on whole brain ischemia in mice. NBP and Borneolum Syntheticum are administered together, and the dosage of NBP is 20, 10 and 5mg/kg-1All can prolong the gasping times of the broken mouse, 20 and 10 mg/kg-1Can prolong the gasp time of the mouse after the head is broken, has significant difference compared with a model group, and suggests that the combined administration of borneol and NBP can enhance the protection effect of NBP on the mouse with ischemia and anoxia, so that the NBP can improve the mouse with ischemia and anoxia at lower dosageThe medicine is used, so that the dosage of NBP is reduced, and adverse reactions are reduced.
TABLE 2 Effect on Whole brain ischemia in mice: (
n＝10)
P <0.05, p <0.01, compared to model groups.
Example 5 Effect on learning and memory behavior after cerebral ischemia-reperfusion in mice
1 animal experiment Material
1.1 drug test drugs: example 1 prepared racemic butylphthalide for injection (NBP) solution, natural borneol solution for injection, racemic butylphthalide for injection + natural borneol solution, model control and sham operation group were administered with 30% HP-beta-CD.
1.2 animals SPF-grade KM mice, male, 18-22 g, provided by the experimental animals center of north river medical university; the production license number of the experimental animal is SCXK (Ji) 2008-1-003. License number for experimental animals: SYXK (Ji) 2011-.
1.3 other Malondialdehyde (MDA) and superoxide dismutase (SOD) kits provided by Nanjing as bioengineering research institute.
1.4 data processing data was analyzed for variance and inter-group testing by SPSS11.5 statistical software.
2 effects on learning and memory behavior after cerebral ischemia-reperfusion in mice
2.1 preparation of mouse cerebral ischemia reperfusion injury model: mouse ip (intraperitoneal injection) 0.4 g-kg of 4% chloral hydrate-1Anaesthetizing, separating bilateral common carotid artery obtusely, except the sham operation group, clamping the common carotid artery on both sides with non-traumatic artery clamp, and taking off the artery clamp after ischemia for 15min (min) for re-perfusion suture. The sham-operated animals suture the skin after blunt dissection of only the bilateral common carotid arteries.
2.2 grouping and administration: 90 Kunming mice are randomly divided into 9 groups, namely model pairsControl (30% HP-beta-CD), sham (30% HP-beta-CD), NBP (20, 10 and 5 mg.kg)-1) NBP + Natural Ice sheet group (NBP 20, 10 and 5 mg. kg)-10.5mg/kg of natural borneol-1) And natural ice tablet group (0.5mg kg)-1) 10 per group. Mice were sutured to the skin and intravenous administration was started with 10 mice per group. The administration was continued for 3 days.
2.3 Effect on mouse diving platform response: and performing a jump table reaction training 30min after the second administration for 24 h. During training, the mouse to be tested is placed in the diving platform instrument and is adaptive to the environment for 5 min. Then electrifying the bottom grid plate, and recording the time (latency time) for the mouse to jump down the plateau and the number of times (error number in the retesting period) for the mouse to receive electric shock from the plateau within 5 min. The method takes the fact that mice can escape to a platform correctly when meeting electrical stimulation as an index. And performing formal test 30min after the third administration for 48h, and recording the latency time of the mice and the error times of the re-measurement period within 5 min.
2.4 effects on the homogenic SOD activity and MDA content of the ischemia-reperfusion mice brains: the mice are killed after completing the diving platform reaction test, brains are taken, 10 times of physiological saline is made into homogenate, the content of MDA is determined by a thiobarbituric acid (TBA) method, the SOD enzyme activity unit in brain tissues is determined by a xanthine oxidation method, and the content of protein is determined by Coomassie brilliant blue.
3 results of the experiment
3.1 Effect on mouse diving platform response: the results show that all indexes of the mice in the model group are very different from those in the sham operation group (P)<0.01), indicating that the model is successful. NBP alone was administered intravenously at 20 and 10mg kg-1The frequency of platform jump errors of the group is obviously reduced, the latency period is obviously prolonged, and the difference is obvious compared with that of a model group, so that the NBP has an obvious intelligence promoting effect. Comparing with model group, Borneolum 0.5mg/kg-1The single intravenous administration has no obvious influence on the error times and the latent period of the mouse diving platform. NBP and borneol are combined, and the dosage of NBP is 20, 10 and 5mg/kg-1The frequency of platform jump errors of the group is obviously reduced, the incubation period is obviously prolonged, and the difference is obvious compared with a model group, which suggests that the memory function of NBP can be enhanced by the combined administration of borneol and NBP, so that the NBP can improve the ischemia and hypoxia effects of mice at lower dose,thereby reducing the dosage of NBP and reducing adverse reaction.
TABLE 3 Effect on learning and memory behavior after cerebral ischemia-reperfusion in mice: (
n＝10)
P <0.05, p <0.01, compared to model groups.
3.2 effects on homogenating SOD activity and MDA content in ischemia-reperfusion mouse brain: the results show that NBP was administered intravenously alone at 20, 10 and 5mg/kg-1The group can obviously inhibit the increase of the MDA content in the brain and the reduction of the SOD activity caused by cerebral ischemia-reperfusion of mice, and has statistical significance compared with the model group. Comparing with model group, Borneolum 0.5mg/kg-1Has no obvious influence on the MDA content and the SOD activity of the brain tissue. NBP and Borneolum Syntheticum are administered together, and the dosage of NBP is 20, 10 and 5mg/kg-1The combination of NBP and borneol has better improvement on the MDA content and SOD activity of brain tissues under the same dosage compared with the single administration of NBP, thereby reducing the effective dosage of NBP, reducing the clinical dosage and reducing the adverse reaction.
TABLE 4 Effect on cerebral ischemia-reperfusion brain tissue SOD and MDA in mice: (
n＝10)
P <0.05, p <0.01, compared to model groups.
Example 6 rat focal cerebral ischemia reperfusion model test
1 animal experiment Material
1.1 drug test drugs: l-butylphthalide (L-NBP) solution for injection, natural borneol solution for injection, L-butylphthalide + natural borneol solution for injection, and 30% HP-beta-CD for model control group and sham operation group were prepared in example 1.
1.2 animals of SPF-level Wistar rat 90 male, 270-300g, provided by the Experimental animal center of North river medical university; the production license number of the experimental animal is SCXK (Ji) 2008-1-003. License number for experimental animals: SYXK (Ji) 2011-.
1.3 data processing data was analyzed for variance and inter-group testing by SPSS11.5 statistical software.
2 protective action on rat focal cerebral ischemia reperfusion
2.1 preparation of focal cerebral ischemia reperfusion model: 270-300g male Wistar rat, fasting and no water supply for 12h, intraperitoneal injection of 10% chloral hydrate (350mg kg)-1) Anaesthetizing, fixing the back, making a median incision in the neck, separating the common carotid artery, ligating the common carotid artery at the proximal end of the common carotid artery, clamping an artery clamp at the distal end, making a V-shaped incision with the diameter of about 2mm between the common carotid artery and the artery clamp, slightly inserting a nylon wire into the common carotid artery from the incision, loosening the artery clamp, entering the internal carotid artery through the bifurcation of the internal carotid artery and the external carotid artery, slowly pushing the nylon wire to the intracranial direction of the internal carotid artery, ensuring that the end of the nylon wire passes through the initial part of the middle cerebral artery to block the entrance of the middle cerebral artery to cause ischemia, and pulling out the tether after 2 hours of ischemia to perform 24 hours of reperfusion. The insertion depth of the tying wire of the sham operation group is about 12.0 +/-0.5 mm, and the rest is the same.
2.2 animals were dosed in groups: wistar rats were randomly divided into 9 groups, i.e., a model control group (30% HP-beta-CD), a sham operation group (30% HP-beta-CD), and an L-NBP group (20, 10, and 5 mg. kg.)-1) L-NBP + Natural Ice sheet set (L-NBP 20, 10 and 5 mg. kg)-10.5mg/kg of natural borneol-1) And natural ice tablet group (0.5mg kg)-1) Each group 10Only. Animals were anesthetized and operated with a single intravenous administration 15min post-surgery.
2.3 neurological deficit score: rats were behaviorally scored 24h post-reperfusion according to Longa5 scoring criteria.
2.4 measurement of cerebral infarction scope: after scoring, the rat is killed by cutting off the head, the brain tissue taken out is rapidly placed in a refrigerator at the temperature of 20 ℃ below zero, the rat is placed in a room temperature environment after 10min, and after the olfactory bulb, the cerebellum and the lower brainstem are cut off, four knives are cut according to the coronal section with the interval of 2mm, and 5 continuous coronal thick sections of the brain are cut. Then, the brain slices were quickly placed in 3ml of 0.2M K containing 1% TTC (2,3, 5-triphenyltetrazolium monochloride)2HPO4/K2HPO4(the solution ratio is 74: 26), incubating at 37 deg.C in dark for 30min, and turning the brain slice once every 5 min. After TTC staining, normal tissue rose red and infarcted tissue was not stained white. The brain slices were then fixed in 10% formaldehyde and the white tissue was carefully dug down and weighed, and the infarcted tissue weight was expressed as a percentage of the cerebral hemisphere to eliminate the effects of cerebral edema.
Cerebral infarction range (%). wt./. wt.. of the operative contralateral hemisphere infarcted portion of the operative lateral hemisphere x 100%
3 results of the experiment
3.1 neurological deficit scoring: the results are shown in Table 5, and the rat nerve function score in the model group is very different from that in the sham operation group (P)<0.01), indicating that the model is successful. L-NBP was administered intravenously at 20 mg/kg-1Compared with a model group, the nerve function score of the group has obvious difference, and the L-NBP is prompted to have obvious cerebral ischemia protection effect. Comparing with model group, Borneolum 0.5mg/kg-1The single intravenous administration has no obvious improvement effect on the nerve defects of rats after ischemia reperfusion. L-NBP and borneol are used together, and the dosage of L-NBP is 20 and 10 mg/kg-1Under the dosage, the nerve function score has obvious difference compared with a model group, and compared with the NBP single administration, under the same administration dosage, the L-NBP and borneol combined administration can enhance the improvement of the L-NBP on the nerve defect of a rat after ischemia reperfusion, so that the L-NBP can achieve the treatment effect at lower dosage, thereby reducing the clinical dosage and the adverse reaction.
3.2 measurement of cerebral infarction scope: the results are shown in Table 4, L-NBP administered alone intravenously at 20, 10 and 5mg/kg-1The dosage can improve the cerebral infarction range of rats after ischemia reperfusion, and the difference is obvious compared with a model group. Borneol 0.5mg/kg-1Compared with a model group, the single intravenous administration has no obvious improvement effect on the cerebral infarction range after the ischemia reperfusion of the rats. L-NBP and Borneolum Syntheticum are administered in combination at dosage of 20, 10 and 5mg/kg in L-NBP-1The dosage can improve the cerebral infarction range of rats subjected to ischemia reperfusion, has obvious difference compared with a model group, and can enhance the improvement of the cerebral infarction range of the rats subjected to ischemia reperfusion by L-NBP through the combined administration of the L-NBP and borneol under the same administration amount compared with the single administration of NBP, thereby reducing the effective dosage of the L-NBP and reducing the clinical dosage of the L-NBP.
TABLE 5 Effect on neurological deficits and extent of cerebral infarction after ischemia-reperfusion in rats: (
n＝10)
P <0.05, p <0.01, compared to model groups.
Example 7 therapeutic Effect on focal cerebral ischemia in rats
1 animal experiment Material
1.1 drug test drugs: l-butylphthalide (L-NBP) liquid medicine oil, natural borneol liquid medicine oil, L-butylphthalide + natural borneol liquid medicine oil prepared in example 2. The model control group and the sham operation control group were soybean oil.
1.2 animals of SPF-level Wistar rat 90 male, 270-300g, provided by the Experimental animal center of North river medical university; the production license number of the experimental animal is SCXK (Ji) 2008-1-003. License number for experimental animals: SYXK (Ji) 2011-.
1.3 data processing data was analyzed for variance and inter-group testing by SPSS11.5 statistical software.
2 therapeutic action on rats with focal cerebral ischemia
2.1 focal cerebral ischemia (MCAO) model preparation: 270-300g male Wistar rat, fasting and no water supply for 12h, intraperitoneal injection of 10% chloral hydrate (350mg kg)-1) Anaesthetizing, fixing the back, making a median incision in the neck, separating the common carotid artery, ligating the proximal end of the common carotid artery, clamping an artery clamp at the distal end, making a V-shaped incision with the diameter of about 2mm between the two, slightly inserting a nylon wire into the common carotid artery from the incision, loosening the artery clamp, entering the internal carotid artery through the bifurcation part of the internal carotid artery and the external carotid artery, slowly pushing the nylon wire to the intracranial direction of the internal carotid artery, ensuring that the end of the nylon wire passes through the initial part of the middle cerebral artery to reach the thinner anterior cerebral artery, ligating the common carotid artery and the nylon wire, disinfecting the operation part by using about 0.1g of penicillin, and suturing the incision layer by layer. The insertion depth of the tying wire of the sham operation group is about 12.0 +/-0.5 mm, and the rest is the same.
2.2 animals were dosed in groups: the postoperative rats were randomly divided into 9 groups, i.e., model control group (soybean oil), sham operation group (soybean oil), and L-NBP group (80, 40, and 20 mg. kg)-1) L-NBP + Natural Ice sheet set (L-NBP 80, 40 and 20 mg/kg)-1Natural borneol 2 mg/kg-1) And natural borneol group (2mg kg)-1) 10 per group. The animals are anesthetized and operated, and the administration is carried out by intragastric administration 15min after the operation and is continuously carried out for 14 days.
2.3 biochemical indexes of blood: 30min after the last administration, rats in each group were bled from the abdominal aorta and the level or amount of superoxide dismutase (SOD) and Malondialdehyde (MDA) in the serum was measured. MDA adopts a thiobarbituric acid method, and SOD activity adopts a xanthine oxidation method.
2.4 measurement of cerebral infarction scope: after scoring, the rat is killed by cutting off the head, the brain tissue taken out is rapidly placed in a refrigerator at the temperature of 20 ℃ below zero, the rat is placed in a room temperature environment after 10min, and after the olfactory bulb, the cerebellum and the lower brainstem are cut off, four knives are cut according to the coronal section with the interval of 2mm, and 5 continuous coronal thick sections of the brain are cut. The brain slices were then quickly placed in 3ml of 0.2M K with 1% TTC2HPO4/K2HPO4(the solution ratio is 74: 26), incubating at 37 deg.C under constant temperature and dark condition for 30min, and turning the brain sheet every 5minOnce. After TTC staining, normal tissue rose red and infarcted tissue was not stained white. Brain slices were then fixed in 10% formaldehyde, white tissue was carefully dug down and weighed, and infarcted tissue weight was expressed as a percentage of the cerebral hemisphere to eliminate the effects of cerebral edema.
Cerebral infarction range (%). wt./. wt.. of the operative contralateral hemisphere infarcted portion of the operative lateral hemisphere x 100%
3 results of the experiment
3.1 Effect on serum SOD Activity and MDA content: the results are shown in Table 6, and the SOD activity of the model rat is obviously reduced, and the MDA content is obviously increased. L-NBP continuous 12d gavage 80 and 40mg kg-1The dosage can obviously improve the SOD activity and reduce the MDA content, and the difference is obvious compared with a model group. Borneol 2 mg/kg compared with the model group-1The single gavage administration has no obvious influence on the SOD activity and the MDA content of MCAO rats, but the borneol and the L-NBP can enhance the improvement capability of the L-NBP on the SOD activity and the MDA content of the serum better by the synergistic administration.
3.2 measurement of cerebral infarction scope: the L-NBP is separately administered by intragastric administration of 80 and 40 mg/kg-1The dose can improve the cerebral infarction range of MCAO rats, and has obvious difference compared with a model group. Borneol 2 mg/kg-1Compared with a model group, the single intragastric administration has no obvious improvement effect on the cerebral infarction range of MCAO rats. The L-NBP and borneol are combined for administration, 80, 40 and 20 mg/kg are administered in L-NBP intragastric administration-1The dosage can improve the cerebral infarction range of MCAO rats, has obvious difference compared with a model group, reduces the effective dosage of L-NBP compared with the single dosage of NBP, and can enhance the improvement of the cerebral infarction range of the rats by the L-NBP by the combined dosage of the L-NBP and borneol under the same dosage, thereby reducing the clinical dosage and the adverse reaction.
TABLE 6 Effect on the serum enzyme index and cerebral infarction Range of MCAO rats: (
n＝10)
P <0.05, p <0.01, compared to model groups.
In conclusion, the butylphthalide can obviously improve blood supply to the brain and/or reduce oxygen consumption of the brain after intravenous administration or oral administration; can obviously improve the nerve motor dysfunction and the damaged memory dysfunction after cerebral ischemia; can antagonize oxygen free radicals, thereby relieving brain injury caused by free radical chain reaction; can obviously reduce the cerebral infarction range. The butylphthalide and the borneol are used together, and compared with the single administration, the effective dosage is reduced; under the same dosage, the treatment effect is obviously enhanced. Therefore, the combination of the butylphthalide and the borneol has a synergistic effect in treating cerebrovascular diseases, particularly ischemic cerebrovascular diseases, can obviously improve the treatment effect of the butylphthalide, reduce the effective dosage of the butylphthalide, reduce the dosage of the butylphthalide, and reduce the occurrence of adverse reactions of the butylphthalide after long-term application.
Claims (12)
1. A pharmaceutical composition comprises butylphthalide, Borneolum Syntheticum and pharmaceutically acceptable adjuvants, wherein the weight ratio of butylphthalide to Borneolum Syntheticum is 10: 1; the auxiliary material is vegetable oil, an antioxidant is optionally added, and the composition is prepared into soft capsules.
2. The pharmaceutical composition of claim 1, wherein: the butylphthalide is selected from one of levo-butylphthalide and racemic butylphthalide.
3. The pharmaceutical composition of claim 1, wherein: the borneol is selected from one of natural borneol, synthetic borneol, (+) -2-borneol with the purity of more than or equal to 98% and (-) -2-borneol.
4. The pharmaceutical composition of claim 3, wherein: the borneol is selected from one of natural borneol and (+) -2-borneol with the purity of more than or equal to 98 percent.
5. The pharmaceutical composition of claim 1, wherein: the vegetable oil is selected from one of oleum Sesami, oleum Maydis, oleum Arachidis Hypogaeae, soybean oil, oleum Armeniacae amarum, oleum Persicae, oleum gossypii semen, oleum Helianthi, and oleum Olivarum; the antioxidant is dibutyl carboxyl toluene.
6. The pharmaceutical composition of claim 5, wherein: the vegetable oil is soybean oil.
7. A pharmaceutical composition comprises butylphthalide, Borneolum Syntheticum and pharmaceutically acceptable adjuvants, wherein the weight ratio of butylphthalide to Borneolum Syntheticum is 20: 1; the auxiliary materials are solvent and cosolvent, and the composition is prepared into injection.
8. The pharmaceutical composition of claim 7, wherein: the solvent is water for injection and ethanol; the cosolvent is cyclodextrin or cyclodextrin derivatives.
9. The pharmaceutical composition of claim 8, wherein: the cosolvent is hydroxypropyl-beta-cyclodextrin.
10. Use of a pharmaceutical composition according to any one of claims 1 to 9 for the preparation of a medicament for the treatment of cerebrovascular disease.
11. Use of a pharmaceutical composition according to any one of claims 1 to 9 for the manufacture of a medicament for the treatment of ischemic cerebrovascular disease.
12. Use of the pharmaceutical composition according to any one of claims 1 to 9 for the preparation of a medicament for the treatment of cerebral infarction.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910315555.8A CN109939102B (en) | 2013-06-13 | 2013-06-13 | Pharmaceutical composition containing butylphthalide and borneol and application thereof |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310231054.4A CN104224772A (en) | 2013-06-13 | 2013-06-13 | A pharmaceutical composition comprising butylphthalide and borneol and applications thereof |
| CN201910315555.8A CN109939102B (en) | 2013-06-13 | 2013-06-13 | Pharmaceutical composition containing butylphthalide and borneol and application thereof |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310231054.4A Division CN104224772A (en) | 2013-06-13 | 2013-06-13 | A pharmaceutical composition comprising butylphthalide and borneol and applications thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109939102A CN109939102A (en) | 2019-06-28 |
| CN109939102B true CN109939102B (en) | 2022-05-17 |
Family
ID=52214257
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910315555.8A Active CN109939102B (en) | 2013-06-13 | 2013-06-13 | Pharmaceutical composition containing butylphthalide and borneol and application thereof |
| CN201310231054.4A Pending CN104224772A (en) | 2013-06-13 | 2013-06-13 | A pharmaceutical composition comprising butylphthalide and borneol and applications thereof |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310231054.4A Pending CN104224772A (en) | 2013-06-13 | 2013-06-13 | A pharmaceutical composition comprising butylphthalide and borneol and applications thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN109939102B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105030760B (en) * | 2014-04-24 | 2018-10-02 | 长弘生物科技股份有限公司 | Stable pharmaceutical composition |
| CN105999279A (en) * | 2016-05-30 | 2016-10-12 | 四川曼赛思医药科技有限公司 | Medicine composition comprising butylphthalide and cosolvent |
| CN107595874B (en) * | 2016-07-12 | 2020-10-30 | 石药集团恩必普药业有限公司 | Pharmaceutical composition containing butylphthalide and application thereof in preparation of drugs treating cerebrovascular diseases |
| CN111346090A (en) * | 2018-12-22 | 2020-06-30 | 江苏先声药业有限公司 | A pharmaceutical composition |
| CN111346089A (en) * | 2018-12-22 | 2020-06-30 | 江苏先声药业有限公司 | Medicinal preparation and preparation method thereof |
| CN114831985A (en) * | 2021-02-02 | 2022-08-02 | 南京宁丹新药技术有限公司 | Application of composition containing riluzole and borneol in preparing medicine for treating cerebrovascular diseases |
| CN114315585B (en) * | 2022-03-04 | 2022-05-13 | 中国医学科学院药用植物研究所 | Hydroxypentyl benzoic acid diester compound, and preparation method and pharmaceutical application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1394880A (en) * | 2001-06-18 | 2003-02-05 | 石家庄制药集团制药技术开发有限公司 | Butylbenzene phthalein cyclodextrin or cyclodextrin derivative clathrate, its preparation method and application |
| CN1605336A (en) * | 2003-10-10 | 2005-04-13 | 中国医学科学院药物研究所 | Application of L-butylphthalide in the process for preparing cerebral infarction preventing and treating medicine |
| CN1623542A (en) * | 2003-12-05 | 2005-06-08 | 中奇制药技术（石家庄）有限公司 | Butyl benzene phthalein soft capsule and its preparation method |
| CN101627992A (en) * | 2004-01-20 | 2010-01-20 | 石药集团中奇制药技术（石家庄）有限公司 | Application of levo butyl phthalide in preparing brain stroke medicament |
| CN102716121A (en) * | 2012-06-06 | 2012-10-10 | 石药集团中奇制药技术（石家庄）有限公司 | Butylphthalide medicine active composition and preparation method of butylphthalide medicine active composition |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2726256B1 (en) * | 1994-10-31 | 1996-12-06 | Vesuvius France Sa | ROLLER ASSEMBLY FOR THE TRANSPORT OF HIGH TEMPERATURE ARTICLES |
-
2013
- 2013-06-13 CN CN201910315555.8A patent/CN109939102B/en active Active
- 2013-06-13 CN CN201310231054.4A patent/CN104224772A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1394880A (en) * | 2001-06-18 | 2003-02-05 | 石家庄制药集团制药技术开发有限公司 | Butylbenzene phthalein cyclodextrin or cyclodextrin derivative clathrate, its preparation method and application |
| CN1605336A (en) * | 2003-10-10 | 2005-04-13 | 中国医学科学院药物研究所 | Application of L-butylphthalide in the process for preparing cerebral infarction preventing and treating medicine |
| CN1623542A (en) * | 2003-12-05 | 2005-06-08 | 中奇制药技术（石家庄）有限公司 | Butyl benzene phthalein soft capsule and its preparation method |
| CN101627992A (en) * | 2004-01-20 | 2010-01-20 | 石药集团中奇制药技术（石家庄）有限公司 | Application of levo butyl phthalide in preparing brain stroke medicament |
| CN102716121A (en) * | 2012-06-06 | 2012-10-10 | 石药集团中奇制药技术（石家庄）有限公司 | Butylphthalide medicine active composition and preparation method of butylphthalide medicine active composition |
Non-Patent Citations (1)
| Title |
|---|
| 冰片药效学研究进展;郭姗姗等;《中国现代中药》;20080630;第10卷(第6期);摘要、第6页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104224772A (en) | 2014-12-24 |
| CN109939102A (en) | 2019-06-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109939102B (en) | Pharmaceutical composition containing butylphthalide and borneol and application thereof | |
| US10265345B2 (en) | Use of extracts from rabbit skin inflamed by vaccinia virus for the manufacture of a medicament for the treatment of acute cerebrovascular disease | |
| BR112020026849A2 (en) | COMPOSITION OF ALGINIC OLIGOSACARIC DIACIDES | |
| CN102688501A (en) | Proanthocyanidin B2 phospholipid compound, and preparation method and application thereof | |
| JP2018503637A (en) | Biphenyl derivatives and uses thereof | |
| CN101530410B (en) | Application of oligomeric proantho cyanidins in preparing products resisting vascular dementia | |
| US20160008414A1 (en) | Pharmaceutical composition containing sceptridium japonicum extract for preventing or treating stroke or degenerative brain diseases | |
| Wan | Nuclear factor κB and inhibitor of κB: acupuncture protection against acute focal cerebral ischemia in rodents | |
| CN104546809B (en) | Application of 3,3 ', 5,5 '-tetra isopropyls-the 4,4 '-bigeminy phenol in preventing and treating cerebral arterial thrombosis | |
| CN107595874B (en) | Pharmaceutical composition containing butylphthalide and application thereof in preparation of drugs treating cerebrovascular diseases | |
| CN102688230A (en) | New applciation of procyanidine B2 | |
| CA2986371A1 (en) | Combination comprising palmitoylethanolamide (pea) and lycopene for use in the treatment of inflammatory diseases | |
| CN102697757B (en) | Application of p-hydroxy benzylidene acetone in preparation of drugs for preventing and/or treating encephalopathy | |
| CN101744806B (en) | Application of pinocembrin raceme in preparation of medicals for cerebral apoplexy | |
| KR101237215B1 (en) | Phamaceutical composition for prevention or treatment of enteritis | |
| Dong et al. | Pharmacological effect of Panax notoginseng saponins on cerebral ischemia in animal models | |
| CN107595875B (en) | Composition containing butylphthalide and application thereof in drugs for treating cerebrovascular diseases | |
| KR20150001109A (en) | A composition for preventing or treating dementia through multi-target control | |
| WO2019114676A1 (en) | New medical use of persimmon leaf extract and of preparation of persimmon leaf extract | |
| CN111346089A (en) | Medicinal preparation and preparation method thereof | |
| CN102028681B (en) | Application of peperphentonamine or salt thereof in preparing drug for preventing/treating encephalopathy | |
| WO2019096144A1 (en) | Use of pientzhuang and preparation thereof in treatment of cerebral stroke sequelae | |
| CN111346090A (en) | A pharmaceutical composition | |
| US20230404945A1 (en) | Application of alpha-asarone in preparation of medicine for preventing or treating hemorrhagic stroke | |
| CN106727605B (en) | Application of cyclovirobuxine D in preparing medicine for preventing or treating cerebral arterial thrombosis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |