CN105168201B - Application of the Vitexin in pain medication after preparing iatrotechnics - Google Patents

Abstract

The present invention relates to pharmaceutical technology field, and in particular to application of the Vitexin in pain medication after preparing iatrotechnics.The pharmaceutical preparation of the present invention is generally used in intraperitoneal injection mode, naturally it is also possible to use other administering modes：(single) acute after modeling administration consumption is 1~10mg/kg, and chronic (repeated multiple times) administration consumption is 3~10mg/kg.The pharmaceutical preparation acute administration and chronic administration of the present invention can effectively mitigate postoperative pain, without calm side effect and tolerance and additive generation be not observed.

Description

Application of vitexin in preparation of medicine for treating postoperative pain

technical field

The invention relates to the technical field of medicine, in particular to the application of vitexin in the preparation of medicine for treating postoperative pain.

Background technique

Pain is an unpleasant sensory and emotional experience associated with tissue damage or potential tissue damage. According to the priority of onset and the duration of the disease, it can be classified into acute pain and chronic pain. Postoperative pain belongs to acute pain (the course of disease is generally less than 6 weeks), and it is a complex physiological response of the body to the disease itself and tissue damage caused by surgery (Zhang Chuanhan. Guidelines for Clinical Pain Treatment [M]. Beijing: China Medical Science and Technology Press, 2008: 502). At present, postoperative pain is a difficult clinical problem. If acute postoperative pain is not handled properly, it will lead to a series of adverse consequences, such as chronic pain, increased chances of incision infection and respiratory and cardiovascular complications, and delayed discharge time. Reduce the quality of life of patients, even lead to physical disability, increase morbidity and mortality (Argoff CE.Recent management advances in acute postoperative pain[J].Pain Pract,2014,14(5):477-87.). Although the pathological mechanism and treatment of postoperative pain have been studied in depth, new analgesic drugs and technologies have been applied in clinic one after another, and various countries have formulated various guidelines for postoperative pain management, but postoperative pain is still under-treated state. A national survey in the United States showed that about 80% of patients experienced acute postoperative pain, and 39% had severe to extreme pain (Apfelbaum JL, Chen C, Mehta SSet al. Postoperative pain experience: results from a national survey suggest postoperative pain continues to be undermanaged [J]. Anesth Analg, 2003, 97(2): 534-40.). At present, the commonly used drugs for the treatment of postoperative pain mainly include opioids, non-steroidal anti-inflammatory drugs (NSAIDs), adjuvants such as anxiolytics and anticonvulsants, and local anesthetics. However, these drugs are often discounted in clinical application due to their limited effectiveness or safety. For example, opioids are currently the most effective analgesic drugs clinically, but their side effects such as respiratory depression, nausea and vomiting, and constipation, as well as their tolerance and addiction limit their clinical application. NSAIDs have only a moderate analgesic effect, and can produce side effects such as peptic ulcer, cardiac function and renal function damage (Argoff CE.Recent management advances in acute postoperative pain[J].Pain Pract,2014,14(5):477 -87.). Therefore, it is also necessary to further search for new drugs for the treatment of postoperative pain with good curative effect, small side effects, and no tolerance and addiction.

Vitexin (vitexin, CAS number: 3681-93-4, chemical name 8-β-D-glucopyranosyl-5,7-dihydroxy-2-(4-hydroxyphenyl)-4H-1-benzopyran-4-one) Vitexin, also known as vitexin, is a natural plant flavonoid carbon glycoside compound. It is one of the main active ingredients of total flavonoids extracted from the dried and mature fruit of the Rosaceae plant Hawthorn (Crateagus pinnatifida), and is also widely present in other plants. Such as vitex, pigeon pea, mung bean, bamboo leaves, nasturtium, passion flower and so on. Its chemical name is 8-β-D-glucopyranose-4', 7-dihydroxyisoflavone, its molecular formula is C ₂₁ H ₂₀ O ₁₀ , and its relative molecular mass is 432.4. The molecular structure is shown in the figure below. Modern pharmacological research has proved that vitexin has various pharmacological effects such as anti-myocardial infarction, lowering blood pressure, anti-inflammatory and analgesic, antibacterial, anti-virus, anti-oxidation, anti-tumor, anti-thyroid, antispasmodic, etc. (Gu Chengbo, Cai Man, Yuan Xiaohan, etc. Research progress on plant resources and pharmacological effects of vitexin[J]. Chinese Journal of Traditional Chinese Medicine, 2015(03):382-89.)

Vitexin chemical structure

At present, there are not many research reports on the analgesic effect of vitexin, and only two foreign literatures clearly show that vitexin, as the only monomeric compound, has certain analgesic effect. Demir et al. (Demir OU, Can OD. Anti-nociceptive effect of vitexin mediated by the opioid system in mice[J]. Pharmacol Biochem Behav, 2013, 109: 23-30.) studies have shown that vitexin can reduce the heat exposure of healthy mice Analgesic effects of physiological responses to injury, mechanical injury, and chemical injury stimuli. The author used three models to show that vitexin has the effect of relieving pain by using three models: the foot licking response induced by hot plate stimulation, the tail flick response induced by mouse tail clip stimulation, and the mouse writhing response induced by intraperitoneal injection of acetic acid . These three animal models all simulate the normal physiological pain state during acute noxious stimuli, and have a clear pain transmission mechanism, which is significantly different from postoperative pain. Postoperative pain mimics the state of persistent pain, which may involve mechanisms such as inflammatory response, peripheral nociceptor sensitization, and central sensitization, but the specific mechanism is still unclear. For example, the dopamine receptor antagonist haloperidol or the kappa opioid receptor agonist U69593 can have anti-nociceptive effects on the above three models, but they are proved to be ineffective analgesics in clinical practice. Therefore, this article cannot confirm that vitexin can reduce postoperative pathological hyperalgesia under surgical conditions.

In addition, Borghi et al. (Borghi SM, Carvalho TT, Staurengo-Ferrari L et al.Vitexin inhibits inflammatory pain in mice by targeting TRPV1, oxidative stress, and cytokines[J].J Nat Prod,2013,76(6):1141-49.) Studies have shown that vitexin has alleviating effects on inflammatory pain induced by capsaicin, formalin, carrageenan and complete Freund's adjuvant. However, inflammatory pain is chronic pain, and its etiology, pain characteristics, and pathogenesis are different from postoperative pain. Clinically, some drugs such as glucocorticoids such as prednisone and dexamethasone are effective for inflammatory pain, but they are not effective for acute sharp pain. , Pain after surgery is invalid. Therefore, it cannot be shown that vitexin can treat postoperative pain. In addition, there is no relevant patent about the application of vitexin in the preparation of analgesic drugs. Therefore, none of the existing data can clearly prove that vitexin as the only active ingredient can be used in the treatment of postoperative pain.

Contents of the invention

In order to solve the current problems of insufficient treatment of postoperative pain, low effectiveness of therapeutic drugs, large adverse reactions or tolerance and addiction, the present invention provides a new clinical application of vitexin, specifically in the preparation of Use of drugs in the treatment of postoperative pain.

The invention provides the application of vitexin as the only active ingredient in the preparation of medicine for treating postoperative pain. Specifically, in the postoperative pain model of mouse hind foot incision, vitexin showed excellent analgesic effect regardless of single administration or repeated administration, and there was no obvious sedative side effect and tolerance .

Technical scheme of the present invention is as follows:

The application of vitexin in the preparation of medicine for treating postoperative pain, wherein vitexin is the only active ingredient used to prepare the pharmaceutical preparation for postoperative pain.

The application of the vitexin described in the present invention in the preparation of medicines for treating postoperative pain, specifically vitexin can relieve or alleviate postoperative pain.

Vitexin described in the present invention can be obtained through commercial means.

The "postoperative pain" described in the present invention is a complex pathophysiological response of the body to the tissue damage caused by the disease itself and the operation, which is manifested as a series of reactions in psychology and behavior and an unpleasant experience in emotion (Zhang Chuanhan . Clinical Pain Treatment Guidelines [M]. Beijing: China Medical Science and Technology Press, 2008: 502).

The "postoperative pain" mentioned in the present invention includes postoperative incision pain, primary disease site pain, and the like.

Mouse rear foot incision postoperative pain animal model described in the present invention has been generally accepted internationally as a model of representative postoperative pain (Pogatzki EM, Raja SN.A mouse model of incisional pain[J]. Anesthesiology, 2003 , 99(4):1023-27.).

The present invention also uses compounds similar in structure to vitexin, such as isovitexin, to carry out the same experiment, and it is found that it has no postoperative analgesic effect.

The pharmaceutical preparations described in the present invention are prepared into common dosage forms in medical pharmacy by adopting conventional preparation methods in the art.

The pharmaceutical preparations are decoctions, granules, tablets, pills or liquid preparations, etc., and in a preferred embodiment of the present invention, they are liquid preparations such as injections, suspension preparations, etc.

A pharmaceutically acceptable carrier is also added to the pharmaceutical preparation.

The pharmaceutical preparation of the present invention is usually used by intraperitoneal injection, and of course other administration methods can also be adopted: after modeling, the acute (single) administration dosage is 1～10 mg/kg (preferably 1, 3, 5.5, 10 mg/kg ), chronic administration (repeated several times), the dosage is 3-10 mg/kg (preferably 3, 5.5, 10 mg/kg) per administration, once a day.

Both acute administration and chronic administration of the pharmaceutical preparation of the present invention can effectively relieve postoperative pain, have no sedative side effects, and do not observe tolerance and addiction.

Description of drawings

Fig. 1 is the establishment of mouse incision pain model, under the premise of no drug administration, the change of postoperative pain threshold of incision pain model (Model) mice and normal control (Control) mice;

Fig. 2 is vitexin sedative effect experiment, the sedative effect of vitexin (3～20mg/kg) to normal mice;

Fig. 3 is the impact of vitexin 1～10mg/kg single administration on the mechanical pain threshold of incision pain model mice, the incision pain model is small after acute (single) administration of vitexin (1～10mg/kg). Changes in the mechanical pain threshold of mice;

Fig. 4 is the impact of vitexin (3～10mg/kg) repeated multiple administrations on the mechanical pain threshold of incision pain model mice; Changes of mechanical pain threshold in incision pain model mice;

detailed description

The implementation of the present invention will be described in detail below in conjunction with the embodiments of the present invention and the accompanying drawings. The following embodiments are implemented on the premise of the technical solution of the present invention, and detailed implementation is provided, but the protection scope of the present invention is not limited Examples described below.

Vitexin used in the following examples was purchased from Aladdin Company with a purity >98% as detected by HPLC.

Example 1 Establishment of mouse incision pain model

test methods:

The establishment method of mouse incision pain model refers to (Pogatzki EM, Raja SN.A mouse model of incisional pain [J]. Anesthesiology, 2003, 99 (4): 1023-27. Sahbaie P, Sun Y, Liang DYet al. Curcumin treatment attenuates pain and enhances functional recovery after incision [J]. Anesth Analg, 2014, 118(6): 1336-44.), the specific establishment method: using an anesthesia machine, mice are anesthetized with 3% isoflurane, and put on anesthesia Face mask, maintain anesthesia with 1.5% isoflurane, and fix in supine position. After the right hind foot of the mouse is sterilized with 75% alcohol, cut the skin with a No. 11 scalpel blade from 0.2 cm close to the heel to the direction of the toe, and make a 0.5 A cm-long longitudinal incision was made to pick out the fascia and cut it longitudinally in the center of the fascia. After hemostasis by light pressure, a stitch was sutured at the midpoint of the incision with a single No. 6 nylon thread, and then the animal was put back into the cage until it woke up. The mice in the control group received anesthesia and aseptic treatment, but no incision operation was performed, and the mechanical pain threshold was measured together with the model group after waking up. The method of measuring mechanical pain threshold refers to (Pogatzki EM, Raja SN.A mouse model of incisional pain [J]. Anesthesiology, 2003, 99(4): 1023-27.) specifically: use Von-Frey fiber silk measurement kit (Stoelting Co., Ltd., U.S.), measured the paw withdrawal threshold (Paw withdrawal threshold, PWT) under a series of Von-Frey fiber filament stimulation of the operation side (right side) hind paw as its mechanical pain threshold, and observed the different groups of mice Time-point changes in mechanical allodynia. Mice were placed alone in a plexiglass grid placed on a metal sieve to adapt to the environment until it was quiet for 3 consecutive days, and the baseline PWT of mice in each group before surgery was measured 3 times one day before surgery, and the average value was taken as the base value . The PWT of the mice on the operated side was measured 2 hours after the operation and then every 24 hours until it returned to the baseline value before the operation, then the measurement was stopped. Von-Frey filaments of 7 strengths were used for determination, and the bending strength was increased approximately logarithmically (equivalent to 0.07, 0.16, 0.4, 0.6, 1.0, 1.4, 2.0g). In the plexiglass grid on the metal screen, adapt to the environment for at least 15 minutes, Von-Frey filament starting from 0.07g, vertically stimulate the paw bottom of the right hind limb of the mouse (the Von-Frey filament is slightly bent as the standard of complete force ), for a duration of ≤4s, observe the reaction of the mouse, if there are phenomena such as paw withdrawal, paw shaking, paw licking, etc., it is regarded as a positive reaction, otherwise it is negative. The fiber filaments of each strength were continuously measured 3 times, and if there were 2 or more responses in the 3 times, it was regarded as PWT. If there are no at least 2 positive reactions, use the adjacent higher level of intensity stimulation. The maximum strength is 2g, if it is greater than this value, it is still recorded as 2g. The minimum force is recorded as 0.07g, and the lower the PWT, the more severe the mechanical hyperalgesia.

Grouping of animals: 24 male ICR mice, 12 mice in the control group (Control), and 12 mice in the model group (Model).

The operation was performed according to the aforementioned "measurement method", and then the mechanical pain threshold was measured. The result is shown in Figure 1. Before the operation (shown as Pre in the figure), there was no significant difference in the mechanical pain threshold (PWT) of the hind paw between the mouse control group and the model group (1.42±0.06g vs 1.28±0.09g, P>0.05). The PWT of the mice in the model group decreased significantly 2 hours after the operation, then gradually recovered, and returned to the level before the operation on the 5th day. Using two-way repeated measures ANOVA, operation factor (F[1,22]=79.49, P<0.0001), time factor (F[6,132]=14.62, P<0.0001) and time × operation interaction (F[6,132] =26.09, P<0.0001) were statistically significant. Bonferroni post hoc analysis was used for comparison between groups. From 2 hours to 4 days after operation, the PWT of mice in the model group was significantly different from that in the control group (compared with the control group, *P<0.05). During the experiment, the blank control group was repeatedly measured with von Frey filaments, but the pain thresholds of the mouse paws remained at the basic level (Figure 1, indicated by gray solid circles). This shows that the mechanical pain threshold of the incision model mice decreased significantly in the period from 2h to 4 days after the operation, indicating that there was significant mechanical hyperalgesia, and then returned to the baseline level before the operation.

Example 2 Vitexin sedative effect experiment

Autonomic activity test method refers to (Zhu Q, Sun, YH; Yun XD et al. Antinociceptive effects of curcumin in a rat model of postoperative pain, Scientific Reports, 2014, 4:4932), specifically: use a tester for autonomic activity in rats and mice (YLS-1B, Jinan Yanyi Science and Technology Co., Ltd., China), the instrument contains a control unit and 4 cylindrical black plastic dark boxes (diameter 30cm, height 30cm), the mouse is put into the black plastic dark, and the light sensor is located in the box The center of the lid is used to record the number of voluntary activities of mice. The number of spontaneous activities of mice within 60 min was recorded.

Animal grouping: 48 male ICR mice; divided into 4 groups, namely the normal control group and vitexin 3mg/kg, 10mg/kg, 20mg/kg three dose groups, 12 in each group. The normal control group was given normal saline, and the remaining groups were injected intraperitoneally with vitexin 3mg/kg, 10mg/kg, and 20mg/kg (prepared with normal saline, and the volume of administration was 10ml/kg), and then observed it within 1h according to the aforementioned measurement method. autonomous activity.

Statistical method adopts one-way analysis of variance, and the comparison between groups adopts Student–Newman–Keuls post hoc analysis, which shows that compared with the normal control group, the voluntary activity count in the 20mg/kg group is significantly lower, and there is a statistical difference (compared with the control group Control, *P<0.05), with a certain sedative effect. However, the 3mg/kg and 10mg/kg dose groups had no significant difference compared with the normal control group, indicating that there was no sedative effect under these two doses (as shown in Figure 2).

Since 20mg/kg has obvious sedative effect, we choose four doses of 1, 3, 5.5, and 10mg/kg for the following acute analgesic experiment.

Example 3 Effect of Acute Administration of Vitexin 1-10 mg/kg on Mechanical Pain Threshold of Incision Pain Model Mice

Grouping of animals: 60 ICR mice, divided into 5 groups; 12 in the vehicle group (given 0.5% CMC-Na); 12 vitexin 1, 3, 5.5, 10 mg/kg dose groups, 12 in each group (Note: administered by intraperitoneal injection, vitexin is configured by 0.5% CMC-Na).

All mice in each group underwent incision surgery, and the operation method was the same as above. The baseline value of PWT in each group was tested 24 hours after the operation (no drug was administered at this time). After administration, the PWT was tested every 30 minutes until the pain threshold returned to the postoperative baseline value. The method of measuring the pain threshold is the same as above. The results are shown in Figure 3. 24 hours after the operation, the baseline value of the mechanical pain threshold (expressed as 0 min) was measured before the administration, and then measured at 30, 60, 90, 120, 150, 180, 210, and 240 min after the administration. Its mechanical pain threshold (PWT). It was found that vitexin at a dose of 3-10 mg/kg could dose-dependently increase the pain threshold of postoperative mice. In the vehicle group (injection of 0.5% CMC-Na intraperitoneally after the operation) within 4 hours of the test period, the repeatedly measured pain thresholds remained at the lowest level (the gray circle in Figure 3). However, the PWT of the mice in the vitexin-administered group gradually increased with time after administration, reached the maximum at 90 minutes, then gradually decreased, and the pain threshold returned to the level before the operation 4 hours after the administration. Using two-factor repeated measures analysis of variance, drug factor (F[4,55]=3.31, P<0.05), time factor (F[8,440]=28.54) and drug × time interaction (F[32,440]=2.66, P <0.0001) are statistically significant. The comparison between groups was analyzed by Bonferroni post hoc. It was found that vitexin at doses of 3mg/kg and 5.5mg/kg could significantly increase the pain threshold of postoperative mice when administered for 90min, while vitexin at 10mg/kg could significantly increase the pain threshold of mice after administration. There is a significant effect from 60 to 150 minutes after taking the medicine (P<0.05). 1mg/kg vitexin had no pain-relieving effect on model mice. In Fig. 3, all the data points filled in black are indicated, which are statistically significant compared with the vehicle group (P<0.05). This suggests that treatment with acutely administered vitexin can dose-dependently improve mechanical hyperalgesia in this model.

Example 4 Effect of chronic administration of 3-10 mg/kg vitexin on mechanical pain threshold of incision pain model mice

Because 1 mg/kg of vitexin has no obvious analgesic effect, we chose three doses of 3-10 mg/kg to conduct the experiment in the experiment of studying the effect of repeated administration of vitexin on incision pain in mice. Grouping of animals: 48 ICR mice, divided into 4 groups; 12 in the vehicle group (giving 0.5% CMC-Na); vitexin 3, 5.5, 10mg/kg three dosage groups, 12 in each group ( Note: Vitexin is administered by intraperitoneal injection, and vitexin is configured by 0.5% CMC-Na). All mice in each group underwent incision surgery, and the operation method was the same as above. The pain threshold was measured 2 hours after operation, and then administered 24 hours after operation, once a day for 6 consecutive days. The pain threshold was measured 90 minutes after administration every day, and the method for measuring the pain threshold was the same as before. The results are shown in Figure 4, daily administration of vitexin can dose-dependently improve mechanical hyperalgesia caused by incision surgery. Using two-factor repeated measures ANOVA, drug factor (F[3,36]=3.56, P<0.05), time factor (F[6,216]=39.94, P<0.0001) and drug × time interaction (F[18,216] =2.20, P<0.01) are statistically significant. The comparison between groups was analyzed by Bonferronipost hoc, 3mg/kg or 5.5mg/kg vitexin can significantly improve the pain threshold of the paw on the first day after surgery, while 10mg/kg dose of vitexin can significantly improve the paw pain threshold 1-3 days after surgery All can significantly reduce the mechanical hyperalgesia of mice (P<0.05). In Fig. 4, all the data points filled in black are indicated, which are statistically significant compared with the vehicle group (P<0.05). This suggests that daily vitexin treatment dose-dependently alleviated mechanical hyperalgesia in this model without developing tolerance.

After the long-term drug administration experiment was completed, the drug was suddenly withdrawn, and no abnormal drug withdrawal reaction was observed in the mice for 3 consecutive days, indicating that it was not addictive.

The above examples are preferred implementations of the present invention, but the implementation of the present invention is not limited by the above examples. Any other modifications made without departing from the spirit and principle of the present invention shall be considered within the protection scope of the present invention.

Claims (7)

1. The application of vitexin in the preparation of medicine for treating postoperative pain, and the postoperative pain is postoperative incision pain. 2. The application of vitexin according to claim 1 in the preparation of medicine for treating postoperative pain, characterized in that, the medicine uses vitexin as the only active ingredient. 3. The application of vitexin according to claim 1 or 2 in the preparation of medicines for treating postoperative pain, characterized in that said medicines are commonly used pharmaceutical preparations in medical pharmacy. 4. The application of vitexin according to claim 3 in the preparation of medicines for treating postoperative pain, characterized in that, said pharmaceutical preparations are decoctions, granules, tablets, pills, injections or suspensions. 5. The application of vitexin according to claim 3 in the preparation of medicine for treating postoperative pain, characterized in that a pharmaceutically acceptable carrier has also been added to the pharmaceutical preparation. 6. The application of vitexin according to claim 3 in the preparation of medicines for treating postoperative pain, characterized in that, the administration method of the pharmaceutical preparation is: after modeling, the acute single administration dosage is 1 ~10mg/kg; or chronically repeated multiple administrations after modeling, the dosage is 3-10mg/kg for each administration, once a day. 7. The application of vitexin according to claim 3 in the preparation of medicines for treating postoperative pain, characterized in that, the administration method of the pharmaceutical preparation is: after modeling, the acute single administration dosage is 1 , 3, 5.5 or 10 mg/kg; or chronic repeated administration after modeling, the medicinal dosage is 3, 5.5 or 10 mg/kg for each administration, once a day.