CN1309968A - Pharmaceutical application of froctose biphosphate magnesium - Google Patents

Abstract

The present invention relates to a new application of magnesium fructose diphosphate, and relates to the application of magnesium fructose diphosphate in the preparation of medicines for treating or preventing central nervous system diseases, and as a component in the preparation of medicines for treating or preventing Alzheimer's disease (senile dementia). Application; application in the preparation of medicine for treating or preventing cerebral ischemic diseases; application in preparation of medicine for treating or preventing epilepsy; application in preparation of medicine for treating or preventing brain injury diseases, etc.

Description

Application of Magnesium Fructose Diphosphate in Pharmaceuticals

The present invention relates to the application of magnesium fructose diphosphate, in particular to the application in the field of pharmacy.

日本专利JP8-81377公开了果糖二磷酸二镁物质，其结构式为： Magnesium fructose diphosphate includes monomagnesium fructose diphosphate and dimagnesium fructose diphosphate. Chinese patent CN95100068.3 discloses monomagnesium fructose diphosphate, and its structural formula is:

Japanese patent JP8-81377 discloses dimagnesium fructose diphosphate, and its structural formula is:

The above two substances can be obtained by fermenting sucrose and phosphate with yeast, adding barium salt to produce fructose-1,6-diphosphate barium, removing barium and adding magnesium salt (the solution is freeze-dried or crystallized); Sodium fructose-1,6-diphosphate can be used to obtain fructose-1,6-diphosphate (FDPH ₄ ) through a cation exchange column, add basic magnesium carbonate to react, filter, and obtain the filtrate after freeze-drying or crystallization.

Magnesium fructose diphosphate is known to be useful as an active ingredient in the treatment of myocardial ischemic diseases such as myocardial infarction, coronary heart disease and the like.

The purpose of the present invention is to provide a new application of magnesium fructose diphosphate, that is, a new application in pharmacy.

The invention mainly relates to the application of magnesium fructose diphosphate as a medicine for treating or preventing central nervous system diseases.

It relates to the application of magnesium fructose diphosphate as a medicine for treating or preventing Alzheimer's disease (senile dementia).

It relates to the application of magnesium fructose diphosphate as a medicine for treating or preventing cerebral ischemic diseases.

It relates to the application of magnesium fructose diphosphate as a medicine for treating or preventing epilepsy.

The invention relates to the application of magnesium fructose diphosphate as a medicine for treating or preventing brain injury diseases.

The inventor found through exploratory experiments that 10-150 mg/kg of magnesium fructose diphosphate can treat and alleviate central nervous system diseases such as Alzheimer's disease, cerebral ischemic disease, epilepsy, and brain injury.

The chemical structure of magnesium fructose diphosphate includes two parts, fructose 1,6-diphosphate and magnesium ion. 1,6-diphosphate fructose can provide energy and improve the ATP deficiency caused by ischemia and hypoxia in the brain; magnesium ions have calcium antagonism, prevent calcium influx, relieve intracellular calcium overload, and regulate intracellular calcium imbalance. Dilates blood vessels in the brain.

Fructose 1,6-diphosphate (FDP) is an important product of glucose metabolism, as well as an important substrate of glucose metabolism, and plays an important regulatory role in many metabolic pathways. One of these is the regulation of glycolysis. Exogenous FDP can avoid glucokinase and PFK inhibited by the acidic environment, directly stimulate pyruvate kinase, and restore the blocked bypass metabolism. Due to the non-energy-consuming two-step phosphorylation process, FDP can produce 1 times more ATP than glucolysis. In addition, FDP can also inhibit the production of oxygen free radicals by neutrophils and reduce their tissue damage.

Magnesium ion is the fourth cation in the human body after Na ⁺ , K ⁺ , and Ca ²⁺ . Its content in cells is second only to K ⁺ . 33% of the magnesium in the human body exists in the cells, most of the rest is deposited in the bone in the form of magnesium phosphate or magnesium carbonate, and only about 1% is located in the extracellular fluid. Magnesium can catalyze or activate 325 enzyme systems; it is a cation necessary for the generation and function of ATP in the body; it participates in the process of oxidative phosphorylation in the body, the production of DNA and RNA, the synthesis of proteins and the structure of all membranes; magnesium and Mg ²⁺ - ATP chelate has a regulatory effect on glycolysis; magnesium is a natural physiological Ca ²⁺ antagonist; it can also maintain the stability of intracellular K ⁺ .

Based on the above mechanism, magnesium fructose diphosphate can improve the ischemic and hypoxic state of central nervous system diseases [mainly including Alzheimer's disease (senile dementia), cerebral ischemic disease, epilepsy, brain injury, etc.], Increase energy supply, supplement magnesium ions, and antagonize neurotoxicity caused by calcium overload. Therefore, it can be used for the treatment or auxiliary treatment of the above diseases.

The medicine made of magnesium fructose diphosphate in the present invention can be injection and oral preparation, and also includes magnesium fructose diphosphate compound preparation containing pharmaceutical additives and/or excipients. In these preparations, the dosage of magnesium fructose diphosphate accounts for 0.1%-100% (weight).

The above-mentioned preparations may also contain other pharmaceutically active ingredients, which can be prepared according to existing methods in a general manner and by adding additives and/or excipients.

The following examples will further illustrate the pharmacological tests and results of magnesium fructose diphosphate, but do not limit the present invention.

Example 1

60 NIH mice were randomly divided into 5 groups: high (30mg/kg), medium (15mg/kg) and low (7.5mg/kg) dosage groups of magnesium fructose diphosphate, Shuangyiping group, and normal saline group. Administration method: ip, qd, for 1 month. Animal models of Alzheimer's disease employ multiple replication models. Test indicators: learning and memory ability, βAP content in the brain. The results in Table 1 and Table 2 show that magnesium fructose diphosphate can significantly improve the learning and memory ability of model mice, reduce the content of βAP in the brain of model mice, and presumably can improve intracellular calcium overload.

Table 1 Effect of magnesium fructose diphosphate on learning and memory of Alzheimer's disease model mice (X±SD) Dose (mg/kg) Number of cases (n) Latency (seconds) Normal control group Normal saline group Shuangyiping group FDP-Mg(l)FDP-Mg(m)FDP-Mg(h) --0.047.51530 151313141314 16.21±5.3252.36±12.5140.54±10.1933.41±11.62 ^* 29.47±9.35 ^* 25.19±12.12 ^*

In the table, ^* means P<0.05 or 0.01 compared with the model control group

As shown in Table 1, the latent period of the Morris water maze of rats was significantly decreased in each experimental group compared with the model control group (P<0.05, P<0.01). It is suggested that FDP-Mg can improve learning and memory.

Table 2 Changes in the number of β-AP positive cells and the average optical density (OD value) in the brains of mice in each group (X±SD) group no Number of positive cells Average optical density CA1 frontal cortex CA1 frontal cortex Normal control group Normal saline group Shuangyiping group FDP-Mg 10101010 8.78±2.05△△ 7.55±2.13△△ 0.093±0.007△△ 0.104±0.008△△37.85 ^± 8.52 ^** 48.07±7.58 ^** 0.136±0.013 ** 0.143±0.018 ^** 35.33±7.37 ^** 44.57±7 ^. 0.134±0.023 ^** 0.141 ±0.017 ^** 18.63±6.30 ^**△ 26.83±8.39 ^**△ 0.112±0.010 ^**△ 0.116±0.014 ^**△

In the table: ^* means P<0.05 or 0.01 compared with normal control group; ^** means P<0.01 compared with normal control group; △ means P<0.05 compared with normal saline group; △△ means P<0.01 compared with normal saline group .

As shown in Table 2, the number of β-AP positive cells in the hippocampal CA1 area and the frontal cortex, and the average optical density (OD) of β-AP immunohistochemical staining were significantly higher in the normal saline group than in the normal control group (P<0.001); The FDP-Mg group was lower than the normal saline group (P<0.05), and the difference between the Shuangyiping group and the normal saline group was not significant (P>0.05). It is suggested that FDP-Mg can reduce the number of β-AP positive cells and the content of β-AP in the interstitium, but Shuangyiping has no obvious effect.

Example 2

The ischemic model of rat four cerebral arteries was ligated. The animals were anesthetized with pentobarbital sodium (40 mg/kg, ip), the bilateral common carotid arteries were isolated, and sutures were placed under them without ligation. Make a median transverse incision on the back of the neck, separate the paraspinal muscles, expose the pterygium foramen of the first vertebra, and heat coagulate. Then the skin was sewn back into the cage, and after 24 hours of free drinking, the bilateral common carotid arteries were clamped with arterial clips in the awake state of the rats. The righting reflex disappeared and the existence of spontaneous breathing was the successful sign of the model. After 40 minutes of ischemia, the carotid clamp was released and reperfused for 1 hour. The results are shown in Table 3.

Table 3 Effects of magnesium fructose diphosphate on cerebral ischemia-reperfusion rats (x±s) group no Brain Water Brain Calcium (%) (μg/gdw ^* ) Lactate dehydrogenase Malondialdehyde (KU/g pro ^△ ) (μmol/g pro ^△ ) Sham operation ischemia reperfusion nimodipine (2mg/kg) magnesium fructose diphosphate (10mg/kg) magnesium fructose diphosphate (20mg/kg) 69677 78.0±1.7c 172±8c82.2±1.2 238±1876.1±1.7c 171±12c80.0±0.5b 146±18c78.1±1.1b 143±9c 20.7±1.5c 14.6±2.6c12.2±1.4 25.6±1.918.9±1.2c 17.2±2.0c18.3±0.5c 16.2±2.6c19.5±1.4c 18.3±1.6c Note: Comparing each group with the ischemia-reperfusion group: b P<0.05, c P<0.01; ^* dw=dry weight; △pro=protein.

The results showed that in the ischemia-reperfusion group, the content of brain water and MDH increased significantly, and the content of LDH in brain tissue decreased (P<0.01); 0.05) and MDA content, increased LDH content (P <0.01). In the ischemia-reperfusion group, the calcium ion content in brain tissue increased significantly (P<0.01), and nimodipine and fructose magnesium diphosphate could significantly decrease the calcium content (P<0.01).

Example 3

Adult Wistar rats, weighing (200-250) g, male or female, were anesthetized with 2.5% pentobarbital sodium (40 mg/kg, ip). Separate the sciatic nerve on one side, infiltrate it with 37℃～38℃ liquid paraffin cotton strips for stimulation, fix the head of the rat with a brain stereotaxic instrument, open 2.5mm behind the bregma and 2.5mm beside the midline as the center, and remove the skull 5mm× 5mm, the dura mater was reserved, and a filter paper (2mm×2mm) impregnated with 5×10 ^-4 mol/L mol/L was applied to this area to create an acute epilepsy model in rats. Insert the stainless steel injection tube into the lateral ventricle (1.5mm posterior to the bregma, 1.5mm lateral to the midline, and 3.5-4mm deep from the skull surface) with the help of a stereotaxic instrument. See the cerebrospinal fluid flow out, confirming that it has been inserted into the ventricle. Inject magnesium fructose diphosphate 20 μg/10 μL into the lateral ventricle, and the injection time is 30-40 seconds. Table 4 is the effect of intracerebroventricular injection of magnesium fructose diphosphate on the amplitude of cortical evoked potential (CEP) in CL-induced epileptic rats (x±s).

Table 4 Effect of intracerebroventricular injection of magnesium fructose diphosphate on the amplitude of cortical evoked potential (CEP) in CL-induced epileptic rats (x±s) Observation items no CEP amplitude before injection/mV CEP amplitude after injection/mV 5min 15min 30min Normal saline group FDP-Mg group 67 1.8±0.361.6±0.61 1.6±0.64 1.60±0.54 1.7±0.631.0±0.35 ^** 0.72±0.15 ^** 0.85±0.05 ^** In the table: ^** indicates P<0.01 compared with the control value.

The results showed that after applying CL for about 10 minutes and injecting magnesium fructose bisphosphate (n=7) into the lateral ventricle, the amplitude of CEP was significantly reduced at about 5 minutes, and the inhibitory effect was most obvious at 15 minutes, and the inhibitory effect could last for more than 30 minutes.

Example 4

Thirty healthy SD rats, half male and half male, weighing 180-220 g, were randomly divided into control group, trauma group and trauma treatment group, with 10 rats in each group. Production of animal models: According to the principle of injury caused by free fall, an impact device is made, which consists of three parts: injury pad, hammer and casing. 2.5% pentobarbital sodium (40mg/kg, ip) anesthetized, fixed the animal on the operating table, sagittal incision of the scalp, exposed the right parietal bone, 2 mm in front of the herringbone suture, 2 mm on the right side of the midline with a desktop A small hole was drilled with a dental drill, and a bone window with a diameter of 5 mm was enlarged with a vascular forceps to keep the dura mater intact. The injury pad was placed on the epidural of the right parietal bone window, and a hammer weighing 20 g was dropped from a height of 30 cm to impact the injury pad to cause local brain contusion of the right parietal lobe. In the trauma treatment group, magnesium fructose diphosphate (150 mg/kg body weight) was intraperitoneally injected immediately after injury. In the normal control group, only the scalp was incised, and a bone window of the same size was opened without causing brain contusion. Animals in each group were fed and watered as usual after awakening from anesthesia, and were sacrificed 24 hours later. Mitochondria were extracted according to conventional methods and confirmed by transmission electron microscopy.

Mitochondrial MDA was measured by pentobarbituric acid (TBA) method, and SOD was measured by xanthine oxidation method. All mitochondrial protein quantifications were determined by the Lowry method.

Determination of mitochondrial Ca ²⁺ and Mg ²⁺ content: Take 1ml of mitochondrial suspension, add 2ml of 1mol/L HNO ₃ , heat in a boiling water bath, then transfer to a 5ml volumetric flask after cooling, dilute to the mark with 8mmol/L Lacl ₃ solution , and then use flame atomic absorption spectrometer to measure mitochondrial Ca ²⁺ , Mg ²⁺ content.

The MDA content and SOD activity in the brain mitochondria of rats in each group are shown in Table 5. The mitochondrial MDA level increased and the SOD activity decreased 24 hours after craniocerebral injury. Compared with the trauma group, the SOD activity in the treatment group was significantly increased, while the MDA content was significantly decreased (P<0.05).

Table 5 The levels of MDA and SOD in brain mitochondria of rats in each group group no MDA (nmol/mg Pr ¹ ) SOD(Nu ² /mg Pr ¹ ) Control group Trauma group Treatment group 101010 15.27±4.1535.54±7.27 ^* 23.62±7.22 ^*# 36.75 ±9.0117.32±6.42 ^* 26.67±6.81 ^*# In the table, ^* means P<0.05 compared with the control group, #P<0.05 compared with the trauma group; 1 means protein; 2 means nitrite unit.

See Table 6 for mitochondrial Ca ²⁺ , Mg ²⁺ content and Ca ²⁺ /Mg ²⁺ ratio in each group.

Table 6 Ca ²⁺ , Mg ²⁺ content and Ca ²⁺ /Mg ²⁺ ratio in brain mitochondria of rats in each group group no Ca ²⁺ (μg/mg Pr) Mg ²⁺ (μg/mg Pr) Ca ²⁺ /Mg ²⁺ Control group Trauma group Treatment group 101010 1.13±0.102.12±0.11 ^* 1.46±0.08 ^*# 0.27±0.04 0.13±0.02 ^* 0.20±0.01 ^*# 4.42±1.02 17.45±3.44 ^* 7.97±0.43 ^*# In the table, ^* indicates P<0.05 compared with the control group, and #P<0.05 compared with the trauma group.

24 hours after craniocerebral trauma, the mitochondrial Ca ²⁺ content increased and the Mg ²⁺ content decreased in the trauma group (P<0.05). Although the Fructose Magnesium Diphosphate treatment group also had the same change trend, the degree of calcium overload and magnesium decreased significantly. The results of Ca ²⁺ /Mg ²⁺ ratio can reflect this change more clearly. Abstract: The present invention provides a new application of fructose diphosphate magnesium (FDP-Mg), that is, the application of FDP-Mg in the treatment of central nervous system diseases, mainly including the treatment of Alzheimer's disease (senile dementia), cerebral insufficiency, etc. Blood disorders, epilepsy, brain damage, etc. FDP-Mg can be used as a therapeutic drug or adjuvant therapeutic drug for the above central nervous system diseases.

Claims (5)

1. Magnesium fructose diphosphate as an application in the preparation of medicine for treating or preventing diseases of the central nervous system. 2. Magnesium fructose diphosphate is used as a medicine for treating or preventing Alzheimer's disease (senile dementia). 3. Application of magnesium fructose diphosphate as a medicine for treating or preventing cerebral ischemic diseases. 4. Application of magnesium fructose diphosphate as a medicine for treating or preventing epilepsy. 5. Application of magnesium fructose diphosphate as a medicine for treating or preventing brain injury diseases.