CN1163104A - Pharmaceutical preparations containing shikimic acid and its derivatives and their application in the preparation of antithrombotic and analgesic drugs - Google Patents

Abstract

The invention relates to a pharmaceutical preparation with shikimic acid and its derivatives as active ingredients. The invention also relates to the new drug application of shikimic acid and its derivatives, specifically the application of shikimic acid and shikimic acid triacetyl derivatives (SA-2) in the preparation of drugs with antithrombotic and analgesic effects.

Description

Pharmaceutical preparations containing shikimic acid and its derivatives and their application in the preparation of antithrombotic and analgesic drugs

The invention relates to a pharmaceutical preparation with shikimic acid and its derivatives as active ingredients. The present invention also relates to new pharmaceutical uses of shikimic acid and its derivatives, specifically the application of shikimic acid and shikimic acid triacetyl derivatives (SA-2) in the preparation of drugs with antithrombotic and analgesic effects, which belongs to the field of pharmaceuticals .

Shikmic acid (Shikmic acid, SA-1) is a medicinal ingredient contained in the fruits of various plants of the genus Illicium in the Chinese herbal medicine Magnoliaceae, and its source is the fruit of Illicium henrvivar. The fruit is extracted by solvents such as methanol and refined. Its molecular formula is C ₇ H ₁₀ O ₅ , and its molecular weight is 174.15. In recent years, the research on the mechanism of thrombosis and the development of antithrombotic drugs have attracted the attention of the medical community in various countries, and some effective compounds are in the clinical and preclinical research stage. Antithrombotic drugs are divided into three categories according to their mechanism of action: anticoagulant drugs, antiplatelet drugs and thrombolytic drugs. Among anticoagulant drugs, in addition to natural and recombinant human antithrombin-III, there are direct thrombin inhibitors hirudin, argatroban, and crude extracts of traditional Chinese medicine Zanthoxylum bungeanum, Salvia miltiorrhiza, and Radix Paeoniae Rubra; antiplatelet drugs are currently A lot of new progress has been made, including drugs rhynchophylline, hollandine A, and paeoniflorol that inhibit the metabolism of arachidonic acid, and tetrandrine, ginkgolide B, and kelptenone that inhibit platelet activating factor, and some Traditional Chinese medicine or ingredients inhibit platelet aggregation by increasing the cAMP level in platelets and reducing the Ca ²⁺ concentration in platelets. With the further elucidation of the pathological mechanism of thrombotic diseases and the mechanism of fibrinolysis in vivo, more and more studies have been conducted on antithrombotic and thrombolytic drugs, which play an important role in the thrombolytic therapy of acute myocardial infarction. It has been found that the aqueous solution of the traditional Chinese medicine Puhuang has a direct fibrinolytic effect independent of the plasminase system. Garlic can significantly reduce plasma fibrinogen, and can directly activate plasmin, so that the fragments produced by decomposing fibrin can be significantly increased. The water extracts of red peony root and earthworm can also produce thrombolysis in different links. In short, the research and development of antithrombotic drugs is very active, and the research on the mechanism of drug action has gone deep into the molecular and receptor levels. Although the current work has achieved a lot of achievements, there are still many problems. The main problem is that the research preparations of traditional Chinese medicine are not pure enough, and there are many impurities in the crude preparations, which have brought great limitations to the efficacy, mechanism of action and clinical administration routes. . Therefore, pure monomer drug is the main direction of future development. At present, the research on shikimic acid at home and abroad is limited to the study of pharmacological effects, and there is no report on shikimic acid pharmaceutical preparations. Pharmacological studies on shikimic acid and its derivatives are mostly focused on cytotoxicity and antibiotics, and there are no studies on the effects on the blood system. In addition, there is no report on other pharmacological effects of shikimic acid such as analgesia.

The purpose of the present invention is to provide a pharmaceutical preparation with good antithrombotic and thrombolytic effects and analgesic effect. The active ingredient is a novel drug monomer with abundant resources, easy extraction, simple structure, and easy artificial synthesis, and has little adverse reaction.

Another object of the present invention is to provide the application of shikimic acid (SA-1) and its derivatives in the preparation of antithrombotic and analgesic drugs.

One of the sources of the medicinal ingredient shikimic acid and its derivatives of the medicine of the present invention is extracted by the following method:

Take the mature pericarp of the star anise plant, crush it, extract it with methanol as a solvent, concentrate, place it, filter the precipitate, treat it with petroleum ether, and repeatedly crystallize methanol to obtain shikimic acid.

Take shikimic acid, add acetic anhydride, then add concentrated sulfuric acid dropwise, let it stand, add the reactant to ice water and shake, let it stand, extract the upper water layer with chloroform, then redissolve the lower reactant in chloroform, wash the chloroform with water, and depressurize Chloroform is recovered to obtain shikimic acid triacetyl.

The medicine of the present invention contains effective amount of shikimic acid and its derivatives, and the shikimic acid derivatives are preferably triacetylshikimic acid.

The medicine of the present invention contains 1%-99% by weight of shikimic acid and its derivatives, and the shikimic acid derivatives are preferably triacetylshikimic acid.

The medicine of the present invention preferably contains 10%-80% by weight of shikimic acid and its derivatives, and the shikimic acid derivatives are preferably triacetylshikimic acid.

The medicine of the present invention more preferably contains 20%-60% by weight of shikimic acid and its derivatives, and the shikimic acid derivatives are preferably triacetylshikimic acid.

The medicine of the present invention optimally contains 25% by weight of shikimic acid and its derivatives, and the said shikimic acid derivatives are preferably triacetylshikimic acid.

The medicament of the present invention also contains pharmaceutical excipients commonly used in the field of pharmacy, such as solvents, fillers, flavoring agents, disintegrants, binders, lubricants, in addition to effective doses of shikimic acid and derivatives thereof. , wetting agent, pigment, solubilizer, thinner, thickener, etc.

According to conventional preparation technology, the medicine of the present invention can be prepared into any pharmaceutical preparation suitable for clinical use, for example, pill, tablet, powder, injection, oral liquid, capsule, suspension, ointment, granule, powder Injections, chewing agents, etc.

The preparation method of pharmaceutical tablet of the present invention is as follows:

Components:

Shikimic acid 25g

Powdered sugar 22 grams

Starch 43 grams

Appropriate amount of 10% starch slurry

Magnesium stearate 0.5 g

Made into 1000 pieces

Preparation method:

Grind shikimic acid and powdered sugar separately, sieve, mix evenly, add starch slurry and stir evenly, the amount of starch slurry added is suitable for granulation, granulate with 16-mesh nylon sieve, dry at 60°C, add Magnesium stearate is mixed and pressed into tablets, each tablet contains 25 mg of shikimic acid.

The preparation method of drug injection of the present invention is as follows:

Components:

Shikimic acid 25g

Sodium chloride 9 grams

Add water for injection to 1000ml

Preparation method:

Take 800 ml of water for injection, add sodium chloride and stir to dissolve it, then add shikimic acid to dissolve it, add water to a sufficient amount, stir, filter, pot, and sterilize with steam at 100°C for 30 minutes to obtain the product.

Compared with the existing antithrombotic drugs, the drug of the present invention has good antithrombotic and thrombolytic effects, and has analgesic, antipyretic, anti-inflammatory and other effects. Moreover, its active ingredient drug monomer is rich in resources, easy to extract, simple in structure, and easy to synthesize or synthesize. Shikimic acid (SA-1) and its derivative triacetylshikimic acid (SA-2) have been proved by pharmacological experiments to have a good antithrombotic effect, and its triacetyl derivatives can strengthen its effect and have a thrombolytic effect. It is a new type of drug for treating thrombotic diseases with good curative effect and small adverse reactions.

Experimental example 1 Experimental research on the antithrombotic effect of shikimic acid 1. The effect of shikimic acid on arterial and venous bypass thrombosis in rats Experimental materials (1) Animals: Wistar male rats weighing 250g to 350g, bred by the Institute of Experimental Animals, Chinese Academy of Medical Sciences Provided at the venue, the certificate number is (Jingdongzi) No. 9201R018. (2) Test drug: The pure product of shikimic acid was provided by Professor Guo Yajian from the Pharmaceutical Chemistry Laboratory of Beijing University of Traditional Chinese Medicine, and stored in a desiccator. When in use, it is freshly prepared with normal saline, adjusted to pH=7 with 1N NaOH solution, and used for intravenous injection. (3) Instrument: LIBRORAEG-220 electronic balance, product of SHMADEE, Japan. Methods and Results Rats were randomly divided into 9 groups, namely the control group, shikimic acid 400mg/kg group, 100mg/kg group, 50mg/kg group, 25mg/kg group, heparin 140μ/rat, ligustrazine 50mg/kg group, 25mg/kg group. Anesthetized by intraperitoneal injection of pentobarbital sodium 50mg/kg, fixed in supine position, neck surgery to separate the right common carotid artery and left external jugular vein. Take three sections of polyethylene pipes (the two sections are 10cm in length, 1mm in inner diameter, and the middle section is 8cm in length, and 2mm in inner diameter), put a 5cm long wire (pre-weighed) into the middle section, and then connect the two sections of thin tubes. Inject heparin saline solution (50 μ/ml) from one end to fill the lumen, insert one end of the tube into the left external jugular vein, and insert the other end into the right common carotid artery. Immediately after the operation was completed, drugs (medication group) and normal saline (control group) were injected into the femoral vein to open the blood flow. After 15 minutes, the blood flow was interrupted, and the silk thread was quickly taken out and weighed. The weight of the thrombus was obtained by subtracting the weight of the thread from the total weight. , baked in an oven at 60°C for 1 hour, weighed, and subtracted the weight of the silk thread to obtain the dry weight of the thrombus. Calculate the inhibition rate according to the following formula.

Thrombus dry (wet) weight in the control group- thrombus dry (wet) weight inhibition rate in the administration group

The dry (wet) weight of thrombus in the control group × 100g

Table 1 Effect of shikimic acid on arteriovenous bypass thrombus in rats Group Dose Number of animals Thrombus wet weight Inhibition rate Thrombus dry weight Inhibition rate

(mg/kg) (n) (mg) (mg) (mg) (%) Control group - 11 25.9±12.2 6.7±3.7 -Shikimic acid 400 10 9.1±4.0 ^** 65 2.8±1.5 58.2 Shikimic acid 200 10 5.9 ±27 ^*** 77 1.6±1.2 76.1 shikimic acid 100 10 6.2±3.8 ^*** 76 1.8±1.2 73.1 shikimic acid 50 10 12.5±8.1 ^** 52 2.4±1.8 64.2 shikimic acid 25 10 8.6±3.3 ^** 67 1.8 ±1.3 73.1 Ligustrazine 50 11 10.9±4.9 ^*** 58 2.4±1.8 64.2 Ligustrazine 25 12 8.6±2.3 ^*** 67 1.8±1.1 73.1 Heparin 140μ/piece 10 2.2±1.0 ^*** 92 0.3±0.3 95.5 ^{* *} and ^*** : Compared with the control group, P<0.01 and P<0.001 Table 1 results show that intravenous injection of shikimic acid 400mg/kg～25mg/kg has anti-arteriovenous circuit thrombosis effect, and the strength of the effect is similar to that of ligustrazine similar. But there is no obvious and regular dose-effect relationship. 2. The influence of shikimic acid on rat common carotid artery thrombosis Experimental materials: (1) Animals: Wistar species male rats with a body weight of 250g～350g, provided by the breeding ground of the Experimental Animal Laboratory of China Medical College, the certificate number is (Beijing Verb) No. 9209-R018. (2) Tested drug: The pure product of shikimic acid was provided by Professor Guo Yajian from the Department of Pharmaceutical Chemistry of our school, and it was stored in a desiccator. Freshly prepared physiological saline, adjusted to pH = 7 with 1N NaOH solution, for intravenous injection. (3) Instrument: BT87-3 type experimental thrombus formation measuring instrument in vivo, developed by Baotou Medical College. Methods and results: Rats were randomly divided into 5 groups and anesthetized by intraperitoneal injection of barbital sodium 50mg/kg. Fix it on the operating table in the supine position, incise the skin of the neck, separate the right common carotid artery 3 cm, place the stimulating electrode of the BT877-3 experimental thrombosis analyzer on the proximal end of the artery, and place the temperature sensing electrode on the distal end of the artery. At the end of the heart, the distance between the two electrodes is 2.5cm. The electrodes were fixed on the operating table. After the operation was completed, shikimic acid solution 400 mg/kg, 200 mg/kg, and 100 mg/kg were injected into the femoral vein in the administration group, and 0.1 ml/piece of 1% heparin solution was given to the heparin group. Give an equal volume of normal saline. After administration, turn on the switch of the thrombosis instrument and give 1.5mν direct current stimulation for 7 minutes to damage the endothelial cells. After 10 minutes, the temperature indicator needle is set to zero. With the formation of thrombus in the cavity, the temperature of the distal end of the artery drops suddenly, and the instrument alarms. Display arterial occlusion time OT (occlusion time), the results are shown in Table 2

Table 2 Effect of intravenous administration of shikimic acid on rat common carotid artery thrombosis Group Dose Zoology Thrombosis time

(mg/kg) (only) OT(X±SD) control group isometric 10 966.1±125.9 shikimic acid group 400 10 1432.2±147.2 ^*** shikimic acid group 200 10 1260.6±190.3 ^*** shikimic acid group 100 10 1362.8 ±127.9 ^*** Heparin group 140U/piece 10 1598.8±271.2 ^*** Note: ^*** Compared with the control group, P＜0.001

The results showed that: compared with the control group, the shikimic acid 400mg/kg group, 200mg/kg group, and 100mg/kg group all significantly prolonged the thrombosis time (P<0.001, P<0.001, P<0.001), and the effects of the heparin group For those who are similar, there is no significant difference between the two groups. 3. Effect of shikimic acid on rat inferior vena cava thrombosis (1) Animal: Wistar male rats, weighing 300-350g, provided by the breeding farm of the Institute of Experimental Animals, Chinese Academy of Medical Sciences, the certificate number is (Jingdongzi) No. 9209-R018. (2) The test drug and other drugs: pure shikimic acid was provided by Professor Guo Yajian from the Pharmaceutical Chemistry Research Office of our school. When used, it was freshly prepared with normal saline, adjusted to pH = 7 with 1N NaOH solution, and used for intravenous injection, with 1% CMC (Sodium carboxymethyl cellulose) is used for intragastric administration after being suspended; the potency of heparin per mg is 140U, batch number 930412, a product of Beijing Purchasing and Supply Station of China Pharmaceutical Company. (3) Instrument: DF206 electric blast drying oven, product of Beijing Jingtong Instrument Factory. Methods and results: ①Intravenous administration of the effect on thrombosis of the ligated inferior vena cava in rats[1].

Rats were randomly divided into 5 groups, anesthetized by intraperitoneal injection of pentobarbital sodium solution 50mg/kg. The supine position was fixed, and the inferior vena cava was separated by laparotomy. It was administered by femoral vein injection. The shikimic acid group was given shikimic acid solution 400 mg/kg, 200 mg/kg, and 100 mg/kg respectively, the heparin group was given 0.1% heparin 0.1 ml per mouse, and the control group was given normal saline with the same volume. After administration, the inferior vena cava was ligated below the branch of the right renal vein of the inferior vena cava, and the vena cava was tied tightly with a No. 5 silk thread. The abdominal wall was sutured, and the abdomen was reopened 6 hours later. The blood vessel was clamped 2 cm below the ligature, the lumen was opened, the thrombus was taken out, and the wet weight was weighed, and then the dry weight was weighed after baking in an oven at 60°C for 1 hour. The results are shown in the table 3.

Table 3 Effect of Shikimic Acid Intravenous Injection on Thrombus Formation of Ligated IVC in Rats Groups Dose Number of Animals Thrombus Wet Weight (X±SD) Thrombus Dry Weight (X±SD)

(mg/kg) (only) (mg) (mg) control group equal volume 8 3.02±8.0 10.0±3.9 shikimic acid group 400 8 33.9±22.0 11.1±7.0 shikimic acid group 200 8 10.6±6.6 ^*** 2.6±1.7 ^*** Shikimic acid group 100 8 19.4±9.7 ^* 5.6±2.8 ^* Heparin group 140U/cat 8 1.8±1.2 ^* 0±0.5 ^* Note: ^*** P＜0.001 ^* P＜0.05

The results showed that: compared with the control group, the shikimic acid 200mg/kg group significantly reduced the dry and wet weight of thrombus (P<0.001), and the shikimic acid 100mg/kg group had a significant effect (P<0.05). Shikimic acid 400mg/kg group had no obvious effect. ②Effect of continuous oral administration on thrombosis of inferior vena cava in rats The rats were randomly divided into 4 groups, and the shikimic acid group was intragastrically administered 1% shikimic acid 1% CMC solution 400 mg/kg and 200 mg/kg once a day; aspirin The group was given aspirin 1% cmc solution 30mg/kg. The control group was given equal volume of 1% CMC. Continuous administration for 3 days, the last administration 60 minutes operation. The influence and method of observing the ligation of inferior vena cava thrombosis are shown in the intravenous administration part of this model, and the results are shown in Table 4. Table 4 The effect of oral administration of shikimic acid on the thrombosis of the ligated inferior vena cava in rats (continuous administration) Groups Dose Number of animals Wet weight of thrombus Dry weight of thrombus

(mg/kg) (only) (mg) (mg) Control group 100 5 20.4±20.5 7.4±7.8 Aspirin group 30 6 45.7±40.0 15.1±12.7 Shikimic acid group 400 4 9.5±6.2 ^** 2.8±1.7 ^** Shikimic acid group 200 5 32.0±22.8 18.0±19.8 Note: ^** P＜0.05

The results showed that the continuous oral administration of 400 mg/kg shikimic acid could inhibit the thrombosis of ligated inferior vena cava (P<0.05), while the oral administration of 200 mg/kg shikimic acid and aspirin had no significant effect. 4. Effect of shikimic acid on mouse model of pulmonary embolism Experimental materials (1) Animals: Kunming male mice weighing 25-30 g, provided by the Experimental Animal Supply Department of Beijing Medical University, certificate number (Jingdongzi) No. 8806M038. (2) Drugs: The pure product of shikimic acid under test was provided by Professor Guo Yajian from the Pharmaceutical Chemistry Research Office of our school. When used, it was freshly prepared with normal saline and 1N NaOH solution pH=7 for the main injection of Jingyong. ADP thrombin is a product of Sigma Company; epinephrine hydrochloride is commercially available. Method and Results

Fifty male mice weighing 25-30 g were randomly divided into 5 groups. Freshly prepare blood coagulation mixture, dissolve 5 mg of thrombin in 5 ml of normal saline; dissolve 1.74 mg of ADP in 2.5 ml of normal saline; then take 0.125 ml of 0.1% epinephrine. Mix the three solutions. The mice were injected into the tail vein respectively, and the shikimic acid group was given shikimic acid solution at 200 mg/kg, 100 mg/kg, and 50 mg/kg. The control group was given equal volume of normal saline. After 1 minute of administration of the above drugs, 0.1 ml/10 g of thrombin mixture was injected into the tail vein of each mouse. The death time and number of mice were recorded, and the death rate was calculated. The results are shown in Table 5.

Table 5 Effect of intravenous administration of shikimic acid on thrombin-induced acute pulmonary embolism in mice

(mg/kg) (only) (only) (only) ( %) (s) The equivalent capacity of the control group is 10 100 100 299.8 ± 116.42 Mangyoxic acid group 10 10 90 313.0 ± 238.08 Mangyoxic acid group 50 10 5 50 623.1 ± 361.93 △ Mang Mango acid Group 100 10 8 80 377.1±319.31Shikimic acid group 200 10 4 40 663.9±313.32△△Note: Significant by chi-square test△,△△P<0.01 compared with control group

The results showed that the mice in the control group were induced with pulmonary embolism, and the mortality rate was 100%, while the groups of 200 mg/kg and 50 mg/kg intravenously injected shikimic acid significantly reduced the mortality rate. The mortality rates of the two were 40% and 50% respectively, and the death time was significantly prolonged (P<0.01, P<0.05). Shikimic acid 100mg/lg, 10mg/kg group had no obvious effect. 5. Effect of shikimic acid on coagulation time Experimental materials (1) Animals: Kunming male mice, weighing 25-30 g, provided by the Laboratory Animal Supply Department of Beijing Medical University, certificate number (Jingdongzi) No. 8806M038. (2) Drugs: Tested drug: pure shikimic acid was provided by Professor Guo Yajian from the Pharmaceutical Chemistry Research Office of our school, and it was freshly prepared with 0.2N NaOH to adjust the pH and then added appropriate amount of normal saline. Methods and results 1. Effect of intramuscular injection of shikimic acid on coagulation time of mice (slide method)

48 male mice were randomly divided into 4 groups. The administration group was injected intramuscularly with 200 mg/kg, 100 mg/kg, and 50 mg/kg of shikimic acid solution, and the control group was given equal volume of normal saline. After 30 minutes, the mouse was quickly cut off at 1cm from the tail tip of the mouse for surgery, and a drop of blood was squeezed on a clean glass slide. The diameter of the blood drop was about 5mm, and the stopwatch was used to time immediately. After 2 minutes, use a clean pin to gently move from the edge of the blood drop inward every 30 seconds to observe whether there is blood streaks formed. When there are blood streaks, it is the coagulation time. The results are shown in Table 6,

Table 6 Effect of intramuscular injection of shikimic acid on blood coagulation time of mice (slide method) group dose number of animals coagulation time

(mg/kg) (only) (X±SD) control group equal volume 12 255±60.7 shikimic acid group 200 12 257±71.0 shikimic acid group 100 12 385±62.4 ^*** shikimic acid group 50 12 360±15.3 ^{** *} Note: ^*** P＜0.001

The results showed that: compared with the control group, the shikimic acid intramuscular injection 100mg/kg group and 50mg/kg group significantly prolonged the coagulation time (P<0.001, P<0.001). The effect of 200mg/kg group was not obvious. 2. Effect of intramuscular injection of shikimic acid on coagulation time of mice (capillary method)

Take 80 male mice, weighing 25-30g, and divide them into 4 groups at random. The administration groups were injected with shikimic acid solution 100 mg/kg and 50 mg/kg intramuscularly, and the Ligustrazine group was injected with Ligusticum Ligusticum solution 100 mg/kg intramuscularly. The control group was given the same amount of normal saline. After 30 minutes, take a capillary glass tube (length 10cm, inner diameter 1mm), insert it into the venous plexus of the mouse medial canthus, and start timing after the blood fills the lumen at a depth of 4-5mm. Take out the capillary glass tube, place it horizontally, After 2 minutes, break off 0.5cm at both ends of the capillary every 30 seconds, and slowly pull it apart to observe whether there are coagulation filaments. When there are coagulation filaments, it is the coagulation time. The results are shown in Table 7.

Table 7 Effect of intramuscular injection of shikimic acid on coagulation time of mice (capillary tube method) group dose number of animals coagulation time

(mg/kg) (only) (X±SD) control group equal volume 20 199.5±38.04 shikimic acid group 1000 20 244.5±46.98 ^** shikimic acid group 50 20 240.0±38.93 ^** tetramethylmethylpyrazine group 100 20 247.5±51.39 ^{* *} Note: ^** P＜0.01

The results showed that: compared with the control group, the shikimic acid 100mg/kg group had a significant effect on prolonging the coagulation time (P<0.01, p<0.001), and the effect was similar to that of ligustrazine, and there was no significant difference between the two groups. 3. Observation of the dose-effect relationship of shikimic acid on blood coagulation time: Experimental materials (1) Animals: Kunming mice, weighing 18-22g, provided by the Experimental Animal Supply Department of Beijing Medical University, certificate number (Jingdongzi) No. 8806M038. (2) The drug under test: the pure product of shikimic acid was provided by Professor Guo Yajian from the Department of Pharmacy and Chemistry of the school. Methods and results 80 mice were randomly divided into 4 groups according to body weight: shikimic acid 8mg/kg, 40mg/kg, 200mg/kg (dose ratio 1:5:25) and normal control group. Injected once, the control group was given the same volume of normal saline for 18 consecutive days. On the 4th day, the 8th day, the 11th day, the 15th day and the 18th day of the administration, the mice were taken from the ocular vein plexus with a capillary tube. Blood, break the capillary every 15 seconds, check whether there is blood coagulation, record the time from blood collection to coagulation, and conduct statistics between groups. The results are shown in Table 8.

Table 8 Dose-effect relationship observation group of shikimic acid on blood coagulation time of mice Dose coagulation time (S, X±SD)

(mg/kg) 4 days, 8 days, 11 days, 15 days, 15 days and 18 days, the control group-98.8 ± 15.1 101.2 ± 40.5 92.6 ± 37.4 103.1 ± 85 103.1 ± 19.9 reckless oxalate 8 113.5 ± 26.4 90.2 ± 75.1 ± 27.2 ± 25.5.5 80.0 ± 27.0 Mango acid 40 158.3 ± 19.3 121.5 ± 34.1 144.9 ± 46.2 161.6 ± 21.1 279.5 ± 84.0 Mango acid 200 177.0 ± 76.8 ± 32.8 104.9 ± 27.2 133.4 216.3 ± 42.0.0

The results in Table 8 show that shikimic acid 8mg/kg has no significant effect on coagulation time. From the 4th day to the 18th day of administration, 40mg/kg prolongs the coagulation time of mice, and the effect is relatively stable. There is no obvious trend that the effect increases with the increase in the number of days of administration. The effect of the 200mg/kg dose group is not stronger than that of 40mg/kg. kg, so no obvious dose-effect relationship was found, and this result needs to be further demonstrated repeatedly. Summary Shikimic acid has anti-thrombotic effect, intravenous injection within the dose range of 25mg/kg-400mg/kg can reduce the wet weight and dry weight of rat arterial and venous circuit thrombus, and the inhibition rate is 52%-77% It also inhibits the formation of venous thrombosis and reduces the dry weight and wet weight of the thrombus formed by ligation of the inferior vena cava. Shikimic acid 50mg/kg～200mg/kg intravenous injection can resist acute mouse pulmonary embolism caused by thrombin, ADP and epinephrine mixture, reduce mortality and prolong death time. Intramuscular injection of 50mg/kg, 100mg/kg and 200mg/kg of shikimic acid can prolong the coagulation time of mice, but there is no obvious dose-effect relationship. Experimental example 2 Experimental research on the analgesic effect of shikimic acid 1. Observation of the analgesic effect of shikimic acid (1) Mouse hot plate method: Experimental materials I. Animals: Kunming female mice weighing 18-22g, which were studied by the Chinese Academy of Medical Sciences Experimental Animal Research Provided by the breeding farm, the certificate number (Jingdongzi) is M139290. II. Test drug: shikimic acid was provided by Professor Guo Yajian from the Department of Pharmacy and Chemistry of our school, prepared with normal saline, and adjusted the pH to 7.0 with 0.5N NaOH, for intramuscular injection; tetrahydropalmatine injection, produced by Guangxi Yinzhou District Pharmaceutical Factory , lot number 9401001. III. Apparatus: hot plate method pain threshold device (small three-purpose water tank, Beijing Xicheng District Medical Equipment Factory; lead box, stopwatch, thermometer). Methods and results A. The mice were placed on a 55 ° I0.5 ° constant temperature water bath hot plate, and the latency period of adding hind feet was recorded as the pain threshold reaction time, and the ones greater than 30 " were discarded, and 60 mice qualified for the pain threshold were taken , divided into 5 groups at random after weighing, normal saline control group, shikimic acid 50mg/kg, 100mg/kg, 200mg/kg group, positive control drug tetrahydropalmatine 15mg/kg group, the route of administration adopts intramuscular injection, the capacity is 0.1ml/20g, before administration, after administration 60', 120', 180', 240', 360' pain threshold time, compared with the control group for inter-group t test, compared with before administration whether there is significance The results are shown in Table 9.

Table 9 Effect of shikimic acid on pain induced by hot plate method in mice Groups Dose Number of animals Before administration Pain threshold time (X±SD)

(mg/kg) (only) after administration 60'120'180'240'360' normal saline 200 12 16.60±2.12 15.79±4.78 20.10±6.00 21.61±8.22 20.27±5.57 20.25±5.47 shikimic acid 200 12 17.27±5.1 33.98±18.26 ^** 39.47±20.04 ^** 31.72±13.98 ^* 43.8±32.52 ^* 29.82±12.70 ^*莽草酸 100 12 20.18±4.39 21.65±8.34 ^* 35.33±23.29 ^* 20.12±5.05 40.96±28.68 ^* 26.80±11.09莽草酸50 12 16.25±3.85 23.25±5.64 ^** 27.32±9.11△△△ 28.50±16.54△ 24.73±6.13△△△ 21.02±10.12 Corydalin 15 12 18.46±3.88 31.38±11.30 ^*** 36.937±14.1 19.14 30.12±16.12 ^*** , ^** and ^* : Compared with the control group, P<0.001, P<0.01 and P<0.05 △△△, △△ and △: Compared with before administration, P<0.001, P<0.01 and P<0.05B. Take 60 mice qualified for the grape threshold. The grouping and administration method are the same as in A. The pain threshold time of the mice is measured at 15', 30', 60', 120' and 180' after administration, and the administration is counted. Whether there is a significant difference between the group and the control group, after administration and before administration. The results are shown in Table 10.

Table 10 Effect of shikimic acid on pain induced by hot plate method in mice Groups Dose Number of animals Before administration Pain threshold time (X±SD)

(mg/kg) (only) after administration 15'30'60'120'180' normal saline 200 12 16.58±4.27 13.70±3.63 15.56±5.49 15.66±5.92 17.00±7.18 16.88±7.37 shikimic acid 200 11 14.47±2.83 17.07±6.33 13.87±3.53 20.13±740△ 19.82±4.21△△△ ^* 29.73±17.61 ^*莽草酸 100 12 16.577±3.99 17.31±752 17.25±5.90 21.81±5.20△ ^* 26.27±10.52 19.37±5.84莽草酸50 12 15.40 ±2.83 18.89±7.13 18.10±6.25 25.68±9.17△△ ^** 29.51±14.52△△ ^** 29.68±12.64△△△ ^** Drytohydropalmatine 60 12 19.14±3.80 40.55±16.56 ^*** 43.82±15.88 ^*** 46.3±16.99 ^*** 34.81±19.51 ^* 32.70±18.74 ^* Remarks: ^*** , ^** and ^* : Compared with buried brine, P<0.001, P<0.01 and P<0.05 △△△, △△ and △: Compared with before administration, P<0.001, P<0.01 and P<0.05

The hot plate experiment proved that shikimic acid 50mg/kg, 100mg/kg, and 200mg/kg all had analgesic effect, and the effect was obvious for 1 hour, and lasted for 4-6 hours. The onset time of action is slower than that of tetrahydropalmatine, but the duration is significantly longer than that of tetrahydropalmatine. C. The effect of oral administration of shikimic acid on the hot plate method in mice 70 mice were randomly divided into 7 groups (see Table 11) and given different doses of shikimic acid aspirin by intragastric administration 1h, 2h after administration , 3h, and 4h, the pain threshold was measured, and the results are shown in Table 11.

Table 11 Effect of shikimic acid on pain induced by hot plate method in mice (X±SD) Mouse pain (S) and dose number pain threshold (S) at different times after drug administration to mice before drug administration 1h 2h 3h 4h control Group 10 12.3±2.1 13.9±7.6 14.8±7.3 20.5±7.9 15.5±9.5 Shikimic acid 0.6g/kg Group 10 15.0±2.8 41.4±17.1△△ ^** 23.9±7.7△△ ^* 20.6±9.1△△ 17.1±5.0 Oxalic acid 0.4g/kg group 10 14.6±3.3 19.4±7.3 18.7±7.4 31.6±9.5 ^** 22.3±9.5△Shikimic acid 0.2g/kg group 10 12.6±3.9 15.2±4.2 20.2±8.4△△ 24.7±11.9△△ 27.8 ±9.3△△Shikimic acid 0.1g/kg group 10 16.1±2.9 16.3±4.4 14.7±4.1 18.6±5.2 21.8±8.3 Aspirin 0.4g/kg group 10 11.0±2.8 14.0±4.4 20.1±10.9△ 14.5±5.2 20.0±10.1 △Aspirin 0.2g/kg group 10 16.1±3.7 20.5±4.8△ ^* 18.9±3.0 ^** 26.0±11.1 28.7±7.7△△ ^** Note: Compared with the control group: ^* P＜0.05, ^** P＜0.01

Compared with before administration: △P<0.05△△P<0.01

The results show that oral administration of 0.6g/kg, 0.4g/kg, and 0.2g/kg of shikimic acid has a certain analgesic effect, and the dose required for the effect is larger than that of intramuscular injection. (2) Mouse acetic acid writhing method experimental materials I. Animals: Kunming mice, half male and half female, weighing 18-22g, provided by the breeding farm of the Institute of Experimental Animals, Chinese Academy of Medical Sciences, certificate number (Jingdongzi) N139290 . II. Drugs: Pure shikimic acid is provided by Professor Guo Yajian from the Department of Pharmaceutical Chemistry of our school. It is ready-to-use, dissolved in normal saline, adjusted to pH = 7 with 0.5N NaOH, and used for intramuscular injection; glacial acetic acid is analytically pure, Beijing Chemical Industry Co., Ltd. Factory products; Rhododendron Injection, produced by Pharmaceutical Factory in Liuzhou, Guangxi, batch number 9401001. Experimental results a. Take 100 mice, half male and half male, and divide them into 5 groups at random after weighing; normal saline group, shikimic acid 200g/kg, 100g/kg, 80g/kg group, tetrahydropalmatine 15mg/kg. Each group was intramuscularly injected according to the dose, and 0.6% acetic acid solution 0.1mg/10g was injected intraperitoneally 1h later, and the number of writhing times of the mice within 0'～10' and 10'-20' were recorded, and the significance comparison between groups was carried out. The results are shown in Table 12.

The results in Table 12 show that intramuscular injection of shikimic acid 200mg/kg, 100mg/kg, and 50mg/kg can all reduce the number of writhing in mice, and the dose effects of 200mg/kg and 50mg/kg are very significant, and 15mg/kg of tetrahydropalmatine has a similar effect. effect. b. Take 86 mice, half male and half female, group them according to Table 12, and administer them by intragastric administration, and the rest of the methods are the same as in the experiment a. The results are shown in Table 13

The results in Table 13 show that oral administration of 0.6mg/kg, 0.4mg/kg, 0.2mg/kg and 0.1mg/kg of shikimic acid to mice has a significant antagonism to the writhing reaction of mice caused by acetic acid, indicating that the analgesic effect is strong , and the effect of aspirin 0.2mg/kg, chlorzoxazone tablet 0.2g/kg is similar.

Table 12 Effect of shikimic acid on pain induced by acetic acid in mice Groups Dose Number of animals Phase times (X±SD)

mg/kg (only) 0'～10'10'～20' normal saline 200 19 28.57±13.86 32.78±10.80 shikimic acid 200 20 16.10±8.35 ^** 21.25±10.72 ^** shikimic acid 100 21 24.00±11.90 27.19±12.37 Shikimic acid 50 19 18.52±7.13 ^** 21.94±10.54 ^** Petatellin 15 19 15.21±12.00 15.21±11.67 ^*** Remarks: P<0.001, P<0.01 and P<0.605 compared with normal saline

Table 13 Effect of shikimic acid on acetic acid-induced writhing in mice (X±SD) Drug animal writhing reaction Times and doses of mice writhing at different times after administration of acetic acid Latency period 0-10min 10-20min Control group 18 298.3±143.7 19.9 ±13.5 28.7±9.6 Shikimic acid 0.6/kg group 10 586.0±310.2 7.2±8.1 9.0±7.1 Shikimic acid 0.4g/kg group 10 394.3±92.8 ^* 5.9±3.6 ^** 9.7±3.9 ^** Shikimic acid 0.2g/kg group 10 584.9±385.9 ^* 6.9±9.0 ^* 9.0±9.1 ^** Shikimic acid 0.1g/kg group 10 671.2±391.4 ^** 3.9±8.7 ^* 7.5±7.4 ^** Aspirin tablets 0.1g/kg group 10 338.6±98.1 5.6±2.8 ^** 10.3±.3 ^** Chlorzoxazone Tablets 0.2g/kg group 18 369.7±172.9 7.4±6.0 ^** 12.1±2.3 ^** Note: Compared with the control group: ^* P＜0.05, ^** P＜0.01 (3) Rat Light Radiation Heat Tail Flick Experiment Material I. Animal: Wistar species, male, body weight 150-220g, provided by the breeding ground of Institute of Experimental Animals, Chinese Academy of Medical Sciences, the certificate number is (Jingdongzi) No. 9209 - No. R018. II. Drug under test: shikimic acid was provided by Professor Guo Yajian from the Department of Pharmacy and Chemistry of our school. It was prepared with normal saline and adjusted to pH 70 with 0.5N NaOH for intramuscular injection. Tetrahydropalmatine injection was produced by a pharmaceutical factory in Yinzhou, Guangxi, batch number 9401001. III. Instrument: Thermal radiation measuring pain threshold device, made by the instrument maintenance room of the school, supervised by the teaching and research section. Experimental method a. Take 50 male rats and divide them into 5 groups at random. Wipe off the sebum with 75% alcohol on the outer 1/3 of each rat’s tail, smear it with charcoal ink, and use 18V regulated current to remove the sebum. The halogen lamp was focused and irradiated on the blackened part of the tail of the rat, and the tail-flick latency was recorded. It was qualified if it was less than 30". kg, 50mg/kg tetrahydropalmatine 15mg/kg. Respectively 30', 60', 120' and 180' after administration, measure the tail-flick latency period of rats, and compare between groups. The results are shown in Table 14. b. Get males 60 rats were randomly divided into 6 groups (see Table 15). In addition to oral administration of various doses, the pain threshold response was measured at 1h, 2h, 3h, and 5h after administration. The method was the same as in experiment a. The results are shown in Table 15.

Table 14 Effect of shikimic acid on light radiation-induced pain in rats Groups Dose Number of animals Before administration Pain threshold time (X±SD)

(mg/kg) (only) after administration 30'60'120'180' normal saline 200 10 6.81±2.44 3.40±0.98 3.88±1.5 23.63±1.97 2.57±0.58 shikimic acid 200 10 7.49±3.08 8.99±5.02 ^** 10.49±4.2 39.26±6.44 ^* 8.28±4.70 ^** shikimic acid 100 10 6.82±1.89 4.89±2.03 7.9±4.10 6.27±4.00 4.50±1.62 ^** shikimic acid 50 10 6.69±2.19 8.11±3.020 ^± 1.62 ^{*** *} 8.69±4.89 ^** 5.92±4.32 ^* Corydalin 15 10 6.67±3.21 6.69±8.60 ^*** 15.29±6.20 ^*** 5.51±2.25 3.70±2.65 Remarks: ^*** , ^** and ^* : with saline Compare P<0.001, P<0.01 and P<0.05

The results in Table 14 show that the rats in the normal saline control group were sensitive to pain after repeated radiation-induced pain, and the tail flick time was between 2.57 seconds and 3.88 seconds. , all had analgesic effect, and there were significant differences compared with the normal saline group at the same time (P<0.05, P<0.01 and P<0.001). The intensity of action is weaker than that of tetrahydropalmatine, but it lasts for a long time.

Table 15 Effect of shikimic acid on tail-flick response to radiation stimulation in rats (X±SD) before drug administration to mice mice pain (S) and dose number pain threshold (S) at different times after administration to mice 1h 2h 3h 4h control Group 10 7.6±24 62±3.7 5.6±26 7.8±4.4 9.1±6.8 Shikimic acid 0.4g/kg Group 10 8.5±3.0 15.4±5.0△△ ^** 15.6±5.1△△ ^** 10.0±4.6 7.7±5.1 shikimic acid 0.2g/kg group 10 7.5±2.9 13 5±5.0△△ ^** 8.2±5.8 10.7±6.2 8.8±6.8 Shikimic acid 0.1g/kg group 10 7.2±3.6 8.5±4.1 7.7±4.6 8.7±4.6 5.9± 4.9 Aspirin 0.4g/kg group 10 7.5±2.8 12.6±5.6△△ ^** 13.4±5.2△△ ^** 12.9±5.3△ 5.5±2.1 Aspirin 02g/kg group 18 6.4±3.4 19.0±3.3△ ^** 15.3±5.1△△ ^** 12.3±6.3△ ^* 8.3±6.6 Note: Compared with the control group: ^* P＜0.05, ^** P＜0.01

Compared with before administration: △P<0.05△△P<0.01 The results in Table 15 prove that oral administration of shikimic acid at 0.4mg/kg, 0.2mg/kg, and 0.1mg/kg can prolong the pain threshold reaction time, of which 0.4 Compared with the control group, the mg/kg and 0.2mg/kg dose groups have significant differences. The effect is similar to that of aspirin and chlorzoxazone, and there is no significant difference between the groups. Summary Mouse hot plate method, mouse acetic acid writhing method, and rat radiant heat-induced pain method proved that shikimic acid 50mg/kg, 100mg/kg, 200mg/kg intramuscular injection and oral administration have obvious analgesic effect, and the action time It can be maintained for more than 6 hours. The intensity of action is similar to that of aspirin and chlorzoxazone tablets. The effect of injection administration is weaker than that of methotrexate, but the action time is longer than that of tetrahydropalmatine. 2. Research on the analgesic characteristics of shikimic acid Experimental materials I. Animals: Wistar male rats, weighing 180-220g, provided by the breeding farm of the Institute of Experimental Animals, Chinese Academy of Medical Sciences, the certificate number is (Jingdongzi) No. 9209-R018 No. Kunming male mice, weighing 18-22 g, provided by the breeding farm of the Institute of Chinese Academy of Medical Sciences, certificate number (Jingdongzi) M139290. II. Drugs: Drugs under test: shikimic acid was provided by Professor Guo Yajian from the Department of Pharmaceutical Chemistry of our school, prepared with normal saline, adjusted to pH 7.0 with 0.5N NaOH for intramuscular injection; sodium pentobarbital, imported from Beijing Purchasing Station of China Pharmaceutical Company Prednisone acetate tablets: produced by Tianjin Lisheng Pharmaceutical Factory, batch number 940806; brewer’s yeast: produced by Beijing Brewery; Antongding: produced by Datong Huida Pharmaceutical Factory, batch number 921229. III. Instrument: MK-ANIMEX Mouse Autonomic Activity Instrument, made in Japan. Methods and results (1) Synergistic test of shikimic acid and pentobarbital sodium; 40 male mice were randomly divided into 4 groups (see Table 16), each group was injected intramuscularly according to the dose, and pentobarbital was injected intraperitoneally after 5' Sodium 30mg/kg, the time between the disappearance and reappearance of the righting reflex was taken as the sleep time, and whether there were differences between the groups was observed. See Table 16.

Table 16 Effect of shikimic acid on hypnosis with pentobarbital sodium Group Dose Number of animals Number of righting reflex disappearance Sleep time (X+S)

mg/kg (only) (only) (only) physiological salt water+pentophyte 200 7 33.43+18.04 reckless oxalic acid+mandarin 200 10 81.75 ± 28.27 reckless oxalic acid+Wu 10 8 26.63 ± 11.07 reckless oxalate+mandarin 50 7 45.43 ±3.68

The above results show that shikimic acid has no obvious synergistic or antagonistic effect on the ophthalmic effect of pentobarbital sodium. (2) The effect of shikimic acid on autonomous activities of mice 110 mice were randomly divided into 5 groups, divided into 5 groups, and injected with normal saline, shikimic acid 200mg/kg, 100mg/kg, 50mg/kg tetrahydropalmatine respectively. 15mg/kg, observe the autonomous activity of mice within 6' in the 60', 180', 360' mouse autonomic activity instrument after administration, the results are shown in Table 17

                                                                                                            ,  time of active activities (6') (X+S)

(mg/kg)  (只)       1h                  3h               6h生理盐水     200     22    741.50±144.60    559.36±167.09    592.00±119.07莽草酸       200     22    743.36±126 89    551.00±173.47    529.32±160.87莽草酸       100     22    786.41±58.75     594.23±130.47    586.91± 78.40 shikimic acid 50 22 733.73±54.66 581.86±145.26 524.95±169.85 tetrahydropalmatine 15 22 618.00±151.52 ± 528.23±117 .117

The above results show that compared with the normal saline group, intramuscular injection of shikimic acid 200mg/kg, 100mg/kg, 50mg/kg, there is no significant difference in the number of spontaneous activities of mice, while giving tetrahydropalmatine group can significantly reduce the number of mouse activities at 60'. activity, and the value was significantly different from that of the normal saline group, and returned to normal at 180', indicating that intramuscular injection of shikimic acid had no inhibitory or exciting effect on the central nervous system. (3) Observation of the antipyretic effect of shikimic acid - the effect on rats with fever induced by brewer's yeast method. Fifty male rats were randomly divided into 5 groups according to body weight, and the physiology of subcutaneous injection of 10% brewer's yeast on the back was measured Saline suspension, rectal temperature was measured after 5 hours, and the temperature increased by about 1°C. Intramuscular injection of normal saline, shikimic acid 200mg/kg, 100mg/kg, 50mg/kg and intraperitoneal injection of positive control drug Antongding 0.15mg/100g, at Rectal temperature was measured 1h, 2h, 3h, and 5h after administration, and statistically significant differences were found between the groups. The results are shown in Table 18.

Table 18 The effect of shikimic acid on the increase of body temperature in rats induced by brewer's yeast Group Dose Number of animals Basis Body temperature after administration (X+S)

(mg/kg) (only) Body temperature 1h 2h 3h 5h before administration Normal saline 200 10 37.10±0.61 38.32±0.61 39.28±0.24 38.31±0.52 38.07±0.48 37.34±0.62 Shikimic acid 200 10 37.23±0.59 38.40±0.2 1.02 ^** 37.56±0.67 ^* 37.47±0.41 37.1±0.71莽草酸 100 10 37.02±0.63 38.44±0.52 37.22±1.01 ^*** 37.83±0.62 37.40±0.43 36.88±0.69莽草酸50 10 37.16±0.73 38.17±0.63 37.95± 0.96 ^*** 37.58±0.78 ^* 37.92±0.54 37.16±0.59 Antidine 105 10 37.10±0.61 38.72±0.63 36.45±0.85 ^*** 36.71±0.83 ^*** 37.02±0.61 37.10±0.69 ^* Remarks: ^*** , ^* and ^* : P<0.001, P<0.001 and P<0.05 compared with normal saline group

The above results show that the three dose groups of intramuscular injection of shikimic acid can significantly reduce the increase in body temperature of rats caused by brewer's yeast at 1h, 2h, and 3h. Continue to rise, while the body temperature of the administration group continues to drop, indicating that intramuscular injection of shikimic acid has an antipyretic effect on rat fever caused by brewer's yeast. (4) Observation of the anti-inflammatory effect of shikimic acid - the effect on paw swelling of rats induced by carrageenan 50 male rats, weighing 180-2200g, were randomly divided into 5 groups, intramuscularly injected with normal saline, shikimic acid 200mg/ kg, 100mg/kg, 50mg/kg, orally give the CMC-Na suspension 40mg/kg of the positive control drug prednisone, and subcutaneously inject 1% carrageenan 0.01mg/kg into the left hind foot after 1h, and measure the 60', 180', 300', 360' rat toe circumference after carrageenan, calculate swelling degree, statistical results are shown in Table 19.

Table 19 Effects of Shikimic Acid on Paw Swelling of Carrageenan Rats Groups Dose Administration Number of Animals Paw Swelling Degree (X±S)

mg/Kg (only) 1h 3h 5h 6h Normal saline 200 Intramuscular injection 10 0.355±0.116 0.671±0.160 0.0945±0.239 0.855±0.202Shikimic acid 200 Intramuscular injection 10 0.180±0.063 ^** 0.565±0.108 0.73 Oxalic acid 100 intramuscular injection 10 0.280±0.149 0.778±0.239 0.785±0.207 0.85±0.211 shikimic acid 50 intramuscular injection 10 0.165±0.136 ^** 0.565±0.164 ^* 0.705±0.248 ^* 0.85±165 oral prednisone 20 40 ^.** 0.300±0.150 ^*** 0.595±0286 ^** 0.575±0.278 ^** Remarks: ^*** , ^** and ^* : Compared with normal saline group, P<0.001, P<0.01, P<0.05 Table 18 shows that intramuscular injection Shikimic acid 200mg/kg, 100mg/kg, and 50mg/kg have certain anti-inflammatory effects on rats with carrageenan-induced acute inflammatory paw swelling, and 200mg/kg and 50mg/kg have more significant effects when administered for 1 hour. Summary The research results on the analgesic effect of shikimic acid show that within the analgesic dose range, there is no central inhibitory effect, and it has certain antipyretic and anti-inflammatory effects.

Example 1

Shikimic acid 25g

Powdered sugar 22 grams

Starch 43 grams

Appropriate amount of 10% starch slurry

Magnesium stearate 0.5 g

Made into 1000 pieces

Grind shikimic acid and powdered sugar separately, sieve, mix evenly, add starch slurry and stir evenly, the amount of starch slurry added is suitable for granulation, granulate with 16-mesh nylon sieve, dry at 60°C, add Magnesium stearate is mixed and pressed into tablets, each tablet contains 25 mg of shikimic acid.

Example 2

Shikimic acid 25g

Sodium chloride 9 grams

Add water for injection to 1000ml

Take 800 ml of water for injection, add sodium chloride and stir to dissolve it, then add shikimic acid to dissolve it, add water to a sufficient amount, stir, filter, pot, and sterilize with steam at 100°C for 30 minutes to obtain the product.

Claims (10)

1, a kind of antithrombotic and analgesic is characterized in that this medicine contains the shikimic acid and the derivant thereof of 1%-99% weight portion.

2,, it is characterized in that this medicine contains the shikimic acid and the derivant thereof of 10%-80% weight portion according to said antithrombotic of claim 1 and analgesic.

3,, it is characterized in that this medicine contains the shikimic acid and the derivant thereof of 20%-60% weight portion according to said antithrombotic of claim 2 and analgesic.

4,, it is characterized in that this medicine contains the shikimic acid and the derivant thereof of 25% weight portion according to said antithrombotic of claim 3 and analgesic.

5,, it is characterized in that described shikimic acid derivative is a SA-2 according to said antithrombotic of arbitrary claim and analgesic among the claim 1-5.

6, shikimic acid and derivant thereof have application in the medicine of anti-thrombosis function in preparation.

7, shikimic acid and derivant thereof have application in the medicine of analgesic activity in preparation.

8,, it is characterized in that described shikimic acid derivative is a SA-2 according to application according to claim 6 or 7 said shikimic acids and derivant thereof.

9, SA-2 has application in the medicine of booster action at preparation antagonism thrombosis and analgesic activity.

10, SA-2 has application in the medicine of thrombus effect in preparation.