WO2015180486A1 - Anti-inflammatory and analgesic drug and preparation thereof - Google Patents

Abstract

The present invention belongs to the technical field of medicine. The present invention relates to a synthesis process of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxyl)ethyl ester and the preparation of a fat emulsion injection thereof. Diclofenac sodium is reacted with 1-bromoethylacetic ester to obtain 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxyl)ethyl ester (referred to as diclofenac ester for short), in the presence of a basic catalyst, the chemical structure of which is as follows. The compound of the present invention is a non-steroidal anti-inflammatory drug, which is prepared into a fat emulsion injection which can be used for intravenous injection. Compared with diclofenac sodium intravenous injectable preparations sold abroad, the diclofenac sodium fat emulsion injection is not only highly safe when used, but also has better anti-inflammatory and analgesic effects in vivo.

Description

Anti-inflammatory and analgesic medicine and preparation thereof Technical field

The invention belongs to the technical field of medicine, and relates to a 1-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester and a synthetic method thereof, and a fat emulsion injection thereof Preparation.

Background technique

Diclofenac sodium (DS), chemical name sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate, belongs to the phenylacetic acid non-steroidal anti-inflammatory drug. Its mechanism of action is to inhibit the conversion of arachidonic acid to prostaglandins (PGs) by inhibiting the activity of cyclooxygenase, thereby producing analgesic and anti-inflammatory effects. At the same time, compared to other non-selective non-steroidal anti-inflammatory drugs, the plasma concentration (IC ₅₀ ) required for inhibition of 50% of cyclooxygenase-2 activity by DS is low, and it shows epoxy to the body in vitro. The similar inhibitory activity of the enzyme-1 is therefore more effective and has fewer side effects. At present, there are many common dosage forms on the market, such as tablets, capsules, pills, and externally used creams, tinctures, gels, suppositories, and injections for intramuscular injection. The preparation is currently widely used in the treatment of various rheumatic, rheumatoid arthritis, neuritis, lupus erythematosus and ankylosing spondylitis. In addition, DS has a wide range of applications in the treatment of acute and chronic arthritis and soft tissue rheumatism, cancer pain, and post-traumatic and post-operative acute pain.

Patients with advanced cancer and post-surgery often experience breakthrough pain, which not only makes the patient intolerable, but also affects the two-way regulation between the nervous, immune, and endocrine systems, inhibits the body's immune function, and affects the disease. Return. In the treatment of breakthrough pain, DS as an analgesic non-steroidal anti-inflammatory drug, often combined with opioids, can significantly improve analgesia without increasing the amount of opioids and side effects. effect. However, the time of breakthrough pain is often uncertain and the duration of pain is also short (30min-60min). Non-intravenous analgesic drugs tend to be slow and bioavailable, which may be needed for the drug to achieve analgesia. At the plasma concentration, the patient's pain has disappeared and the analgesic effect is not obvious (Anderson S K, Al Shaikh B A. Diclofenac in combination with opiate infusion after joint replacement surgery [J]. Anaesthesia and intensive care, 1991, 19 (4 ): 535-538.) (DeNatale C E, Rosenberg A, Gharibo C. Rapid acting analgesics [J]. Techniques in Regional Anesthesia and Pain Management, 2010, 14(2): 65-74.), and intravenous preparations It is often able to directly reach the concentration required for drug action, so the development of DS as a safe, stable and rapid onset of intravenous preparations may become a better choice for patients who have experienced breakthrough pain when taking medications. DS is prone to some adverse reactions at normal doses, mainly gastrointestinal irritation, while Basotti et al found that DS at therapeutic doses Parenteral administration does not affect the motor function of the upper gastrointestinal tract as oral administration, so gastrointestinal adverse reactions caused by intravenous DS are milder than oral administration (Bassotti G, Bucaneve G, Betti C, et al. Effects of parenteral diclofenac sodium on upper gastrointestinal motility after food in man. European journal of clinical pharmacology, 1991, 41 (5): 497-500), plus for some gastrointestinal surgery can not Oral, or oral administration of non-steroidal anti-inflammatory drugs may also be administered intravenously, so the development of DS intravenous preparations is of great significance.

ampoule 75mg/3ml)，由于双氯芬酸钠溶解度不高，

在处方中采用大量丙二醇和苯甲醇对DS进行增溶，可用于静脉注射(Mazurek S，Szostak R.Quantitative determination of diclofenac sodium and aminophylline in injection solutions by FT-Raman spectroscopy[J].Journal of pharmaceutical and biomedical analysis，2006，40(5)：1235-1242.)。但是该制剂在临床使用时存在以下问题：

由于采用有机溶媒溶解DS，在静脉注射前需要用大量的碱性生理盐水进行稀释，同时其需要缓慢的进行静脉输注来减少静脉炎的发生率，一般需要30-120分钟以上(详见

ampoule说明书)，所以患者药物使用的顺应性与安全性均较差。Taking into account the above requirements, Novartis developed diclofenac sodium injection (

Ampoule 75mg/3ml), due to the low solubility of diclofenac sodium,

DS is solubilized by a large amount of propylene glycol and benzyl alcohol in the prescription, which can be used for intravenous injection (Mazurek S, Szostak R. Quantitative determination of diclofenac sodium and aminophylline in injection solutions by FT-Raman spectroscopy [J]. Journal of pharmaceutical and biomedical Analysis, 2006, 40(5): 1235-1242.). However, the preparation has the following problems in clinical use:

Due to the use of organic solvents to dissolve DS, it is necessary to dilute with a large amount of alkaline saline before intravenous injection, and it needs slow intravenous infusion to reduce the incidence of phlebitis, generally takes 30-120 minutes or more (see

Ampoule instructions), so the patient's drug use is less compliant and safe.

Chinese patent CN200810101867.0, (Publication No. CN101244278A) discloses an intravenous preparation containing hydroxypropyl-β-cyclodextrin for inclusion of DS, which improves the solubility of DS in water and reduces the direct injection of DS. Vascular irritancy, but we have to point out that in recent years there have been a large number of reports in the literature that intravenous hydroxypropyl-β-cyclodextrin has nephrotoxicity and bone loss (Liu Wei, Cheng Xiaoxiang, Guo Chuanmin, et al. Experimental study on toxicity of hydroxypropyl-β-cyclodextrin administered to rats by abdominal cavity and vein[J].Drugs & Clinic,2009,24(6):361-364.)(Rosseels M L A,Delaunois A G, Hanon E, et al. Hydroxypropyl-β-cyclodextrin impacts renal and systemic hemodynamics in the anesthetized dog [J]. Regulatory Toxicology and Pharmacology, 2013, 67(3): 351-359.) (Kantner I, Erben R G. Long -Term Parenteral Administration of 2-Hydroxypropyl-β-Cyclodextrin Causes Bone Loss [J]. Toxicologic pathology, 2012, 40 (5): 742-750.), so DS injections containing hydroxypropyl-β-cyclodextrin are when using it However, a certain safety problems, and how to develop a more safe and effective DS intravenous injection, is still an urgent problem to solve.

At present, the technique of encapsulating poorly soluble drugs in fat milk for intravenous administration has received a lot of attention from researchers. Clinically, there are also a large number of intravenous drug-loaded fat emulsion preparations, such as propofol medium long-chain fat emulsion, dexamethasone palmitate injection, etc., which not only improve the solubility of the drug, but also reduce it. The irritancy of the drug during intravenous injection also improves the therapeutic effect of the drug (Zhao Mingming, Su Min, Tang Xing. Research progress of drug-loaded fat emulsion injection [J]. Journal of Shenyang Pharmaceutical University, 2010, 27 (12): 1014-1022.) (Yokoyama K, Watanabe M. Limethason as a lipid microsphere preparation: An overview[J]. Advanced drug delivery reviews, 1996, 20(2): 195-201.) (Wang Yun, Yu Wenjun, Zhang Wen , et al. Clinical observation of propofol medium long-chain fat emulsion injection intravenous anesthesia injection pain [J]. Shaanxi Medical Journal, 2010 (2): 238-239.). So the preparation of DS as a fat emulsion preparation may be a good DS intravenous preparation, but DS and diclofenac have lower solubility in soybean oil or other approved intravenous oils, if they are directly made into fat emulsions. The drug loading will be very low. For example, Cui Yue et al studied the diclofenac fat emulsion preparation, the DS fat emulsion prepared by using more oil phase (20%, mass fraction) has a drug content of only 2.5 mg/ml, and the formulation also contains a co-emulsifier. Tween-80 and poloxamer 188 (Cui Yue. Study on diclofenac sodium lipid microsphere injection [D]. Shenyang Pharmaceutical University, 2009.), and there are reports that Tween-80 has serious consequences Sensing effect (Luo Xia, Wang Qing, Zhou Lian, et al. Study on the relationship between Tween-80 and allergic reaction in traditional Chinese medicine injections [J]. Journal of Adverse Drug Reactions, 2010, 12 (3): 160-165.), Therefore, the preparation needs to be improved in terms of safety in use. The clinically required dose of diclofenac sodium is large (75 mg). If the DS fat emulsion of the above study is used for administration, at least 30 ml or more is needed, so that a large volume of the preparation requires a long infusion time, delaying the town. Pain effect Fluctuation also affects patient compliance. In addition, it also increases the amount of excipients in terms of drug cost, increases production costs, increases the price of drugs, and increases the economic burden on patients. Currently, DS fat emulsion preparations have not been listed.

Summary of the invention

The purpose of the present invention is to overcome the defects of the prior art, and to modify the structure of DS to develop a non-steroidal anti-inflammatory drug. The compound has a fat solubility which is much higher than that of DS, and is prepared into a fat emulsion, and the drug loading amount can be large. It is safer to use and safer. Its anti-inflammatory and analgesic effects are better than those of DS intravenous injections.

The main technical scheme of the present invention is to modify the structure of diclofenac sodium (DS) to obtain a new compound of 1-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester ( Diclofenacate, a derivative of the non-steroidal anti-inflammatory drug diclofenac sodium. Further studies have shown that the synthesis of diclofenac salt with diclofenac sodium is simple and the drug yield is high. The synthesized diclofenac ester has good oil solubility and can be used for preparing fat emulsion preparation for intravenous injection. The drug loading is high, safe and efficient.

According to a first aspect of the present invention, there is provided a 1-(acetyloxy)ethyl 2-((2,6-dichlorophenyl)amino]phenylacetate (referred to as diclofenac), which has the following structural formula :

According to a second aspect of the present invention, there is provided a method for synthesizing the above diclofenac, the synthetic route of the method being:

That is, diclofenac sodium is reacted with 1-bromoethyl acetate in the presence of a catalyst which is substituted for the reaction conditions and a basic catalyst to synthesize the compound of the present invention.

The alkaline catalyst may be selected from one or both of potassium hydrogencarbonate, sodium hydrogencarbonate, sodium carbonate, potassium carbonate and the like.

Preferably, the compound diclofenac ester of the present invention is prepared by using 1 mol of the raw material sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate as a feeding unit, and the steps are as follows:

1. 1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate was added to the reaction vessel, dissolved in an organic solvent, and a catalyst was added thereto, followed by stirring at room temperature.

The organic solvent used in the reaction is preferably one or more of N,N-dimethylacetamide, N,N-dimethylformamide, and acetone. The reaction is carried out by adding 0.5 to 1 L of an organic solvent, preferably 0.8 L.

The reaction is carried out by adding a basic catalyst in an amount of from 0.1 to 1.2 mol, preferably from 1 to 1.1 mol. The mixture is stirred at room temperature for 0.5 to 4 hours, preferably for 1 to 1.5 hours.

2. 1.1 to 2.0 mol of 1-bromoethyl acetate is added dropwise to the reaction solution, and the reaction is carried out at room temperature for a while, and then the temperature is raised to a certain temperature to carry out a reaction.

The amount of 1-bromoethyl acetate added in the reaction is 1.1 to 2.0 mol, preferably 1.2 mol. The reaction time at room temperature is 5 to 30 minutes, preferably 20 minutes. The temperature in the temperature rising reaction stage is from 30 ° C to 80 ° C, preferably 55 ° C. The reaction time after the temperature rise is 0.5 to 8 hours, preferably 4 to 5 hours.

3. After the reaction was stopped, the organic solvent and water were added to the reaction mixture, and the organic phase was separated. After washing with water, the mixture was washed with a sodium thiosulfate aqueous solution and finally washed with saturated brine.

In the extraction step, one or more of ethyl acetate, diethyl ether and methyl tert-butyl ether are selected as the extractant. The total amount of solvent added is from 0.5 to 5 L, preferably from 2 to 3 L. The volume ratio of the extractant to water is from 1:1 to 1.5, preferably 1:1.3. The pH of the aqueous sodium thiosulfate solution is from 9 to 11, preferably from 10.2 to 10.5.

4. The organic phase is rotated to evaporate the organic solvent, and the crude product is dried under vacuum to diclofenac.

In a third aspect of the invention, there is provided a fat emulsion injection containing the above compound, which overcomes the technical drawbacks of the presence of the marketed diclofenac sodium injection.

The diclofenac ester provided by the invention can be used as a stable fat emulsion for intravenous injection with high drug loading. The intravenous fat emulsion has a high degree of targeting effect, and can accumulate drugs in the inflammatory site during metabolism in the body, and has a better anti-inflammatory effect. The diclofenac ester emulsion in the diclofenac fat emulsion is dispersed in the form of lipid microspheres. After the high temperature sterilization, the average particle size is in the range of 170-300 nm, and can be directly intravenously injected without dilution with physiological saline, and is convenient to use. No hemolysis and irritation.

The invention provides a diclofenac fatty acid injection, which is composed of diclofenac ester, injectable oil, refined lecithin, glycerin, pH adjuster and water for injection, and the ratio of each component is (the following percentage refers to relative to In terms of the total volume of the injection).

A diclofenac fatty acid injection according to the present invention, wherein the diclofenac is the main drug, and the amount thereof is 0.5 to 2% g/ml, preferably 0.8 to 1.5% g/ml, more preferably 1% g/ ML.

A diclofenac fatty acid injection of the present invention, wherein the injection oil is selected from the group consisting of a mixture of soybean oil for injection or soybean oil for injection and caprylic acid triglyceride for injection, in an amount of 10% or 20%. In grams per milliliter, the preferred amount is 10% gram per milliliter.

A diclofenac ester fat emulsion injection according to the present invention, wherein the injection oil is selected from the group consisting of a mixture of soybean oil for injection, soybean oil for injection and caprylic acid triglyceride for injection, wherein the soybean oil for injection is The content of the mixture shall not be less than 50% by weight (g/g).

A diclofenac ester fat emulsion injection according to the present invention, wherein the pH adjuster is selected from one or two of citric acid, hydrochloric acid, phosphoric acid, acetic acid, disodium hydrogen phosphate, and sodium hydroxide.

A diclofenac ester fat emulsion injection of the present invention, wherein the pH adjuster has a pH of from 6.0 to 8.0, preferably from 6.5 to 7.5.

A diclofenac ester fat emulsion injection of the present invention, the preparation method comprising the following steps:

1. Weigh an appropriate amount of injectable oil, refined egg yolk lecithin, mix well, heat in a water bath to 55 ° C ~ 80 ° C, stir or shear to dissolve, then add diclofenac ester at 55 ° C ~ 80 ° C stir or cut The solution was dissolved to obtain an oil phase.

In this procedure, in which lecithin is dissolved in the oil phase, lecithin can also be dispersed in the aqueous phase.

2. Disperse glycerin in an appropriate amount of water for injection and heat to 55 ° C to 80 ° C to obtain an aqueous phase.

3. The oil phase is slowly injected into the water phase at 55 ° C to 80 ° C while maintaining high shear for 10 to 15 minutes to obtain colostrum.

4. The mixed emulsion is further emulsified by a high pressure homogenizer through a homogenizing pressure of 10,000 to 15,000 psi, and the volume is adjusted to a pH of 6.0 to 8.0 with a pH adjusting agent.

5, filtration, sub-package, nitrogen filling, sealing, sterilization 121 ° C, sterilization for 15 minutes, that is, a diclofenac fat emulsion injection.

In this procedure, a capsule filter having a pore size of 0.45 μm or 0.8 μm was used for filtration.

A diclofenac ester fat emulsion injection of the invention has an average particle diameter of 170 to 300 nm.

The diclofenac ester fat emulsion injection of the invention is a terminal fat emulsion injection, which is directly intravenously injected during clinical use, and does not require a liquid dispensing process, thereby avoiding the risk of cross-contamination and facilitating clinical medication.

The invention not only provides a novel diclofenac ester and a synthetic method thereof, but also successfully prepares a safe, high-efficiency and high drug-loading diclofenac ester fat emulsion injection. The diclofenac ester of the invention has higher fat solubility than diclofenac sodium, and has better anti-inflammatory and analgesic effects in vivo than diclofenac sodium; the synthetic route has the following advantages:

1. The synthetic raw material is an industrialized product, and the cost is low and the performance is stable.

2. The synthetic route is simple, and the requirements for production personnel and production equipment are low.

3. The synthesized product has high purity and high yield, and is suitable for mass production to meet industrial production demand.

The intravenous fat emulsion prepared by the compound of the invention shows that the preparation has rapid onset and remarkable effect, and is safer to use than the commercially available intravenous diclofenac sodium solution, and is anti-inflammatory and town. The pain effect is better and has a long-lasting effect.

DRAWINGS

Figure 1 is a ¹ H NMR spectrum of diclofenac ester.

Figure 2 is a MS (ESI) spectrum of diclofenac ester.

Figure 3 is a graph showing the hemolytic test of diclofenac fat emulsion injection.

Figure 4 is an experimental tissue section of diclofenac sodium injection and diclofenac fat emulsion injection, in which 1-diclofenac sodium injection group; 2-normal saline control group; 3-diclofenac ester fat emulsion injection group; Blank emulsion control group.

detailed description

The present invention will be further described in detail below with reference to the accompanying drawings and embodiments, but the invention is not limited thereto.

Example 1: Preparation Example of Compound of the Present Invention

1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate was added to the reaction vessel, 1 L of N,N-dimethylacetamide was added to dissolve, and 1.1 mol of sodium carbonate was added as a catalyst at room temperature. After stirring for 2 hours, 1.2 mol of 1-bromoethyl acetate was added dropwise, and the mixture was reacted at room temperature for 10 minutes, then heated to 40 ° C, and reacted for 4 hours, and then cooled to room temperature; 3 L of a mixed solvent of ethyl acetate and water (acetic acid) was added. Ethyl acetate: water 1:1), the organic phase was separated, washed with 3 L of water, washed with 3 L of sodium thiosulfate aqueous solution (pH=10), and finally washed with 3 L of saturated brine; The crude product and the crude product were dried under vacuum for 10 hours, and then 278.46 g of a viscous oily substance was obtained, which was obtained as a target product, diclofenac-1-acetoxyethyl ester, and the yield was 92.0% with respect to the raw material diclofenac sodium.

The ¹ H NMR and MS (ESI) spectra of the viscous oily substance are shown in Figures 1-2, respectively, and the corresponding data are as follows:

¹ H NMR (CDCl ₃ , 600 MHz, ppm) δ: 1.50 (d, 3H, J = 5.4 Hz, OCH CH ₃ ), 2.06 (s, 3H, CH ₃ CO), 3.81 (d, 1H, Ph CH _A H _B CO, J = 15.0 Hz), 3.86 (d, 1H, PhCH _A H _B CO, J = 15.0 Hz), 6.57 (m, 1H, Ar-H), 6.64 (br s, 1H, NH), 6.91 (q) , 1H, J=5.4Hz, O CH CH ₃ ), 6.98 (m, 2H, Ar-H), 7.14 (m, 1H, Ar-H), 7.23 (m, 1H, Ar-H), 7.34 (m , 2H, Ar-H).

MS (ESI ⁺ ) m/z: 404 (M+Na ⁺ , 100%).

Example 2: Preparation Example of Compound of the Present Invention

1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate was added to the reaction vessel, 0.5 L of N,N-dimethylformamide was added and dissolved in 0.3 L of acetone, and 1.2 mol of carbonic acid was added. Potassium was used as a catalyst. After stirring at room temperature for 4 hours, 1.5 mol of 1-bromoethyl acetate was added dropwise. After reacting for 30 minutes at room temperature, the temperature was raised to 80 ° C. After 8 hours of reaction, it was cooled to room temperature; 5 L of ethyl acetate was added. A mixed solvent of water (ethyl acetate:water 1.5:1), the organic phase was separated, washed with 5 L of water, washed with 5 L of sodium thiosulfate aqueous solution (pH=11), and finally washed with 5 L of saturated brine; Evaporation of the organic solvent gave a crude oily product. The crude product was dried in vacuo for 24 hours. After the organic solvent was removed, 266.54 g of a viscous oily substance was obtained, and the viscous oily substance was identified as the target product diclofenac by ¹ H NMR and MS (ESI). The yield of 1-acetoxyethyl ester relative to the raw material diclofenac sodium was 90.0%.

Example 3: Preparation Example of Compound of the Present Invention

Add 1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate to the reaction vessel, add 0.5 L of N,N-dimethylacetamide and dissolve in 0.3 L of acetone, and add 1 mol of sodium carbonate. As a catalyst, after stirring at room temperature for 1 hour, 1.1 mol of 1-bromoethyl acetate was added dropwise, and the mixture was reacted at room temperature for 20 minutes, and then heated to 50 ° C. After 5 hours of reaction, it was cooled to room temperature; 4 L of ethyl acetate and water were added. The mixed solvent (ethyl acetate: water 1.2:1), the organic phase was separated, washed with 4L of water, washed with 4L aqueous sodium thiosulfate (pH = 10.2), and finally washed with 4 L of brine; After the organic solvent was removed, the crude product was obtained as a viscous oil. The crude product was dried under vacuum for 8 hours, and then the residual organic solvent was obtained to obtain 268.02 g of a viscous oily substance. The viscous oily substance was identified as the target product diclofenac by ¹ H NMR and MS (ESI) spectra. The yield of 1-acetoxyethyl ester relative to the raw material diclofenac sodium was 90.5%.

Example 4: Preparation Example of Compound of the Present Invention

1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate was added to the reaction vessel, dissolved in 0.8 L of acetone, and 0.5 mol of sodium hydrogencarbonate was added as a catalyst, and the mixture was stirred at room temperature for 0.5 hour. 1.1 mol of 1-bromoethyl acetate was added dropwise, and the mixture was reacted at room temperature for 5 minutes, and then the temperature was raised to 30 ° C. After 0.5 hour, the reaction was cooled to room temperature; 3 L of a mixed solvent of diethyl ether and water (ether: water: 1.5:1) was added and separated. The organic phase was washed with 3 L of water, washed with 3 L of sodium thiosulfate aqueous solution (pH=9), and finally washed with 3 L of saturated brine; and the organic phase was evaporated to remove the organic solvent to give a crude oily product. After the residual organic solvent, 210.27 g of a viscous oily substance was obtained. The white solid was identified as the target product diclofenac-1-acetoxyethyl ester by ¹ H NMR and MS (ESI) spectrum, and the yield of diclofenac sodium was 71.0. %.

Example 5: Preparation Example of Compound of the Present Invention

1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate was added to the reaction vessel, 0.8 L of N,N-dimethylacetamide was added thereto, and 0.1 mol of potassium hydrogencarbonate was added as a catalyst. After stirring at room temperature for 1.5 hours, 1.1 mol of 1-bromoethyl acetate was added dropwise, and the mixture was reacted at room temperature for 20 minutes, and then heated to 55 ° C. After 4.5 hours of reaction, it was cooled to room temperature; 0.5 L of ethyl acetate, diethyl ether, and A was added. A mixed solvent of tert-butyl ether and water (mixed organic solvent: water: 1.3:1), the organic phase was separated, washed with 0.5 L of water, washed with 0.5 L of sodium thiosulfate aqueous solution (pH = 10.5), and finally 0.5 washed with L saturated brine; to give a viscous oily crude organic phase is rotary evaporated off the organic solvent, the crude product was dried in vacuo nine hours in addition to an organic solvent to give a viscous oily substance 181.54g, by ¹ HNMR and MS (ESI) spectra confirmed the The viscous oily substance was the target product diclofenac-1-acetoxyethyl ester, and the yield based on the raw material diclofenac sodium was 61.3%.

Example 6: Preparation Example of Compound of the Present Invention

Add 1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate to the reaction vessel, add 0.4 L of N,N-dimethylacetamide, 0.3 L of N,N-dimethyl Acetylamine and 0.3 L of acetone were dissolved, 0.5 mol of sodium carbonate and 0.5 mol of potassium carbonate were added as a catalyst, and after stirring at room temperature for 1 hour, 1.1 mol of 1-bromoethyl acetate was added dropwise, and the mixture was heated at room temperature for 20 minutes and then heated. After reacting for 4 hours at 55 ° C, it was cooled to room temperature; a mixed solvent of 0.5 L of methyl t-butyl ether and water (methyl tert-butyl ether: water 1.3:1) was added, and the organic phase was separated and washed with 2 L of water. Washed with 2 L of sodium thiosulfate aqueous solution (pH = 10.2), and finally washed with 2 L of saturated brine; the organic phase was evaporated to remove the organic solvent to obtain a crude oily product. The crude product was dried under vacuum for 8 hours, and then the organic solvent remained. The oily substance was 275.72 g, and it was confirmed by ¹ H NMR and MS (ESI) spectrum that the viscous oily substance was the target product diclofenac-1-acetoxyethyl ester, and the yield of the raw material diclofenac sodium was 93.1%.

Example 7: Preparation Example of Compound of the Present Invention

Add 1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate to the reaction vessel, add 1 L of N,N-dimethylacetamide, and add 0.5 mol of sodium carbonate and 0.5 mol of potassium carbonate. And 0.2 mol of potassium hydrogencarbonate as a catalyst, stirred at room temperature for 1.5 hours, 1.2 mol of 1-bromoethyl acetate was added dropwise, the reaction was carried out at room temperature for 20 minutes, and then the temperature was raised to 55 ° C, and the reaction was allowed to cool for 5 hours, and then cooled to room temperature; 2L of a mixed solvent of ethyl acetate and water (ethyl acetate: water: 1.3:1), the organic phase was separated, washed with 2L of water, washed with 2L of sodium thiosulfate aqueous solution (pH = 10.2), and finally with 2 L of saturated brine The organic phase was evaporated to dryness to give a crude oily residue. The crude product was dried under vacuum for 10 hrs and then evaporated to dryness to afford 274.53 g of viscous oily material, which was confirmed by ¹ H NMR and MS (ESI). The target product diclofenac-1-acetoxyethyl ester had a yield of 92.7% relative to the raw material diclofenac sodium.

Example 8: Preparation Example of Compound of the Present Invention

1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate was added to the reaction vessel, 1 L of N,N-dimethylacetamide was added, and 0.4 mol of potassium hydrogencarbonate and 0.4 mol of carbonic acid were added. After stirring at room temperature for 1.5 hours, sodium hydrogenate was added dropwise 1.3 mol of 1-bromoethyl acetate. After reacting for 15 minutes at room temperature, the temperature was raised to 35 ° C. After 2 hours of reaction, it was cooled to room temperature; 2 L of ethyl acetate was added. a mixed solvent with water (ethyl acetate: water: 1.3:1), the organic phase was separated, washed with 2L of water, washed with 2L aqueous sodium thiosulfate (pH = 10.2), and finally washed with 2 L of brine; The crude oil was obtained by rotary evaporation of the organic solvent. The crude product was dried in vacuo for 10 hours, and then the residue was evaporated to give 247.29 g of viscous oily material. The viscous oily material was confirmed to be the target product by ¹ H NMR and MS (ESI). The yield of diclofenac-1-acetoxyethyl ester was 83.5% relative to the raw material diclofenac sodium.

Example 9: Preparation Example of Compound of the Present Invention

1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate was added to the reaction vessel, 1 L of N,N-dimethylacetamide was added to dissolve, and 1.1 mol of sodium carbonate was added as a catalyst at room temperature. After stirring for 3 hours, 1.1 mol of 1-bromoethyl acetate was added dropwise, and the mixture was reacted at room temperature for 15 minutes, then heated to 50 ° C, and reacted for 4.5 hours, and then cooled to room temperature; 2 L of a mixed solvent of ethyl acetate, diethyl ether and water was added. (mixed organic solvent: water 1.3:1), the organic phase was separated, washed with 2 L of water, washed with 2 L of sodium thiosulfate aqueous solution (pH = 10.4), and finally washed with 2 L of saturated brine; to give the crude product as a viscous oil, the crude product was dried in vacuo nine hours after the addition of residual organic solvent to give a viscous oily substance 270.12g, by ¹ HNMR and MS (ESI) spectra confirmed the desired product as a white solid 1-acetoxy diclofenac The yield of ethyl ester was 91.2% relative to the raw material diclofenac sodium.

Example 10: Preparation Example of Compound of the Present Invention

1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate was added to the reaction vessel, 1 L of N,N-dimethylacetamide was added to dissolve, and 1 mol of potassium hydrogencarbonate was added as a catalyst at room temperature. After stirring for 1 hour, 1.2 mol of 1-bromoethyl acetate was added dropwise, and the mixture was reacted at room temperature for 20 minutes, and then heated to 55 ° C. After 4 hours of reaction, it was cooled to room temperature; 2.5 L of ethyl acetate and methyl t-butyl group were added. a mixed solvent of ether and water (mixed organic solvent: water 1.5:1), the organic phase was separated, washed with 2.5 L of water, washed with 2.5 L of sodium thiosulfate aqueous solution (pH = 11), and finally with 2.5 L of saturated brine. After washing, the organic phase was evaporated to dryness of the organic solvent to give a crude oily product. The crude product was dried under vacuum for 7 hrs, and then the organic solvent was left to give 259.72 g of viscous oily substance. The viscous oil was confirmed by ¹ H NMR and MS (ESI). The substance was the target product diclofenac-1-acetoxyethyl ester, and the yield based on the raw material diclofenac sodium was 87.7%.

Example 11: Determination of oil-water partition coefficient of diclofenac and diclofenac sodium

Diclofenac ester can be used in any of the samples prepared in any of Examples 1-10;

Diclofenac sodium can be purchased from Jiangsu Suzhou Fifth Pharmaceutical Factory Co., Ltd.;

pH 7.4 phosphate buffer or pH 3.0 citrate buffer mixed with n-octanol in a volume ratio of 1:1, 1:2, 2:1, and placed in a constant temperature water bath shaker At 25 ° C, 100 rpm, shake for 3 days to equilibrate. An appropriate amount of diclofenac sodium raw material or diclofenac ester was dissolved in the above saturated n-octanol-buffer system, placed in a constant temperature water bath shaker, kept at 25 ° C, 100 rpm, shaken for 2 days to dissolve the distribution equilibrium. The solution was allowed to stand at 25 ° C for 24 hours, and the n-octanol phase and the buffer liquid phase were separated, filtered through a 0.22 μm filter, and the drug content in the two phases was tested by a high performance liquid phase method. The logP value of the oil-water partition coefficient is calculated as follows:

Logp=log (drug solubility in n-octanol/drug solubility in buffer)

The experimental results are shown in Table 1:

Table 1 Determination of oil-water partition coefficient of diclofenac and diclofenac sodium

Compound Logp in phosphate buffer Logp in citrate buffer Diclofenac sodium 0.91 4.03 Diclofenac 4.63 4.43

The experimental results show that the diclofenac ester of the new compound is much more soluble than diclofenac sodium.

Example 12: Example of pharmaceutical preparation of the invention--Selection of type and amount of oil for injection

The drug-loaded fat emulsion is prepared, provided that the main drug has a certain amount of solubility in the oil for injection. Otherwise, a stable drug-loaded fat emulsion cannot be prepared. The diclofenac ester fat emulsion of the present invention is intended to be a long-term storage drug-loaded fat emulsion, so it is necessary to ensure that the selected injectable oil has sufficient solubility for diclofenac.

Weigh the appropriate amount of diclofenac, and place it in 70 ° C soybean oil for injection, caprylic acid triglyceride and a mixture of the two (weight ratio of 1:1) to make 50 mg / g, 80 mg / g, The concentration of 100 mg/g, 120 mg/g, 150 mg/g, 180 mg/g, and 200 mg/g was stirred with heating. The solution was observed after the dissolved drug solution was stored under refrigeration at 4 to 6 ° C for 72 hours.

In soybean oil, when the drug concentration reached 150 mg/g, the solution was slightly turbid, and the solubility of the drug in soybean oil was about 120 mg/g. In the mixture of the two, when the drug concentration reached 180 mg/kg, it was turbid, indicating that the solubility of the drug in the mixture was about 150 mg/g. In the caprylic acid triglyceride, when the drug concentration reached 200 mg/kg, it was slightly turbid, indicating that the solubility of the drug in the mixture was 180 mg/g.

Diclofenac has a solubility of 120 mg/g in soybean oil for injection and a greater solubility in caprylic acid triglyceride, indicating good fat solubility. The commercial intravenous diclofenac sodium is 75 mg/3 ml, and its clinical single dose is 75 mg. Usually, the drug-loaded fat emulsion has an oil content of 10% (g/ml). In summary, both the injectable soybean oil and the caprylic acid triglyceride can be used as an injectable oil to prepare a diclofenac ester fat emulsion.

At present, the fat emulsion for clinical use is mostly 10% or 20% g/ml. Considering the clinical dose, the type and safety of the oil for injection, we use 10% or 20% of the oil for injection. /ml.

Example 13: Example of pharmaceutical preparation of the invention--determination of drug loading

Based on the solubility of the above diclofenac ester, the oil content of the injection and the dose of the clinical drug, we have determined that the drug loading of the diclofenac ester emulsion is diclofenac 5 mg/ml to 20 mg/ml.

Example 14: Example of a pharmaceutical preparation of the present invention

0.5 g of diclofenac ester, 10 g of soybean oil for injection, and 1.2 g of refined egg yolk lecithin were weighed, mixed, placed in a water bath and heated to 55 ° C, and stirred or sheared to dissolve to obtain an oil phase. The glycerin was dispersed in an appropriate amount of water for injection and heated to 55 ° C to obtain an aqueous phase. The oil phase was slowly injected into the aqueous phase at 55 ° C while maintaining high shear for 10 min to obtain colostrum. This mixed emulsion was passed through a high pressure homogenizer, emulsified 3 times under a pressure of 15,000 psi, made up to 100 ml with water for injection, and adjusted to a pH of 6.01 with hydrochloric acid. It was filtered with a 0.8 μm pore size capsule filter, subpacked, nitrogen-filled, sealed, and sterilized at 121 ° C for 15 min to obtain a diclofenac fat emulsion injection.

The drug loading was determined to be 5 mg/ml; the average particle diameter was 170.3 nm; and the pH was 5.55. In accordance with the quality requirements of the present invention, it can be directly administered intravenously when used clinically.

Example 15: Example of a pharmaceutical preparation of the present invention

0.8 g of diclofenac ester, 10 g of soybean oil for injection, and 1.2 g of refined egg yolk lecithin were weighed, mixed, placed in a water bath and heated to 64 ° C, and stirred or sheared to dissolve to obtain an oil phase. The glycerin was dispersed in an appropriate amount of water for injection and heated to 64 ° C to obtain an aqueous phase. The oil phase was slowly injected into the aqueous phase at 64 ° C while maintaining high shear for 10 min to obtain colostrum. The mixed emulsion was passed through a high pressure homogenizer, emulsified 3 times under a pressure of 14,000 psi, made up to 100 ml with water for injection, and adjusted to a pH of 6.32 with hydrochloric acid. It was filtered with a 0.8 μm pore size capsule filter, subpacked, nitrogen-filled, sealed, and sterilized at 121 ° C for 15 min to obtain a diclofenac fat emulsion injection.

The drug loading was determined to be 8 mg/ml; the average particle diameter was 171.7 nm; and the pH was 5.89. In accordance with the quality requirements of the present invention, it can be directly administered intravenously when used clinically.

Example 16: Example of a pharmaceutical preparation of the present invention

1 g of diclofenac ester, 10 g of soybean oil for injection, and 1.2 g of refined egg yolk lecithin were weighed, mixed, and heated to 74 ° C in a water bath, and stirred or sheared to dissolve to obtain an oil phase. The glycerin was dispersed in an appropriate amount of water for injection and heated to 74 ° C to obtain an aqueous phase. The oil phase was slowly injected into the aqueous phase at 74 ° C while maintaining high shear for 10 min to obtain colostrum. The mixed emulsion was passed through a high pressure homogenizer, emulsified 3 times under a pressure of 11,000 psi, made up to 100 ml with water for injection, and adjusted to a pH of 6.11 with hydrochloric acid. It was filtered with a 0.8 μm pore size capsule filter, subpacked, nitrogen-filled, sealed, and sterilized at 121 ° C for 15 min to obtain a diclofenac fat emulsion injection.

The drug loading was determined to be 10 mg/ml; the average particle diameter was 185.7 nm; and the pH was 5.58. In accordance with the quality requirements of the present invention, it can be directly administered intravenously when used clinically.

Example 17: Example of a pharmaceutical preparation of the present invention

1 g of diclofenac ester and 10 g of soybean oil for injection were weighed, mixed, placed in a water bath and heated to 74 ° C, and stirred or sheared to dissolve to obtain an oil phase. The glycerin was dispersed in an appropriate amount of water for injection, 1.2 g of refined egg yolk lecithin was added, mixed, and heated to 74 ° C to obtain an aqueous phase. The oil phase was slowly injected into the aqueous phase at 74 ° C while maintaining high shear for 10 min to obtain colostrum. The mixed emulsion was passed through a high pressure homogenizer, emulsified 3 times under a pressure of 10,000 psi, made up to 100 ml with water for injection, and adjusted to a pH of 6.10 with hydrochloric acid. It was filtered with a 0.8 μm pore size capsule filter, subpacked, nitrogen-filled, sealed, and sterilized at 121 ° C for 15 min to obtain a diclofenac fat emulsion injection.

The drug loading was determined to be 10 mg/ml; the average particle diameter was 193.4 nm; and the pH was 5.65. In accordance with the quality requirements of the present invention, it can be directly administered intravenously when used clinically.

Example 18: Example of a pharmaceutical preparation of the present invention

1 g of diclofenac ester, 5 g of soybean oil for injection, 5 g of medium chain oil for injection, and 1.2 g of refined egg yolk lecithin were weighed, mixed, placed in a water bath and heated to 74 ° C, and stirred or sheared to dissolve to obtain an oil phase. The glycerin was dispersed in an appropriate amount of water for injection and heated to 74 ° C to obtain an aqueous phase. The oil phase was slowly injected into the aqueous phase at 74 ° C while maintaining high shear for 10 min to obtain colostrum. The mixed emulsion was passed through a high pressure homogenizer, emulsified 3 times under a pressure of 10,000 psi, made up to 100 ml with water for injection, and adjusted to a pH of 6.51 with hydrochloric acid and sodium hydroxide. It was filtered with a 0.8 μm pore size capsule filter, subpacked, nitrogen-filled, sealed, and sterilized at 121 ° C for 15 min to obtain a diclofenac fat emulsion injection.

The drug loading was determined to be 10 mg/ml; the average particle diameter was 200.3 nm; and the pH was 6.05. In accordance with the quality requirements of the present invention, it can be directly administered intravenously when used clinically.

Example 19: Example of a pharmaceutical preparation of the present invention

2 g of diclofenac ester, 10 g of soybean oil for injection, and 10 g of caprylic acid triglyceride for injection were weighed, and the mixture was heated to 80 ° C in a water bath, stirred or sheared to dissolve, and an oil phase was obtained. The glycerin was dispersed in an appropriate amount of water for injection, 1.2 g of refined egg yolk lecithin was added, mixed, and heated to 80 ° C to obtain an aqueous phase. The oil phase was slowly injected into the aqueous phase at 80 ° C while maintaining high shear for 15 min to obtain colostrum. The mixed emulsion was passed through a high pressure homogenizer, emulsified 3 times under a pressure of 10,000 psi, made up to 100 ml with water for injection, and adjusted to pH 8.00 with hydrochloric acid and sodium hydroxide. It was filtered with a 0.45 μm pore size capsule filter, subpacked, nitrogen-filled, sealed, and sterilized at 121 ° C for 15 min to obtain a diclofenac fat emulsion injection.

The drug loading was determined to be 19.9 mg/ml; the average particle diameter was 195.4 nm; and the pH was 7.62. Quality in accordance with the invention For clinical use, it can be administered directly intravenously.

Example 20: Example of a pharmaceutical preparation of the present invention

2 g of diclofenac ester, 1.2 g of refined egg yolk lecithin, and 20 g of soybean oil for injection were weighed, and the mixture was heated to 75 ° C in a water bath, and stirred or sheared to dissolve to obtain an oil phase. The glycerin was dispersed in an appropriate amount of water for injection, mixed, and heated to 75 ° C to obtain an aqueous phase. The oil phase was slowly injected into the aqueous phase at 75 ° C while maintaining high shear for 15 min to obtain colostrum. The mixed emulsion was passed through a high pressure homogenizer, emulsified 3 times under a pressure of 10,000 psi, made up to 100 ml with water for injection, and adjusted to a pH of 7.03 with disodium hydrogen phosphate. It was filtered with a 0.22 μm pore size capsule filter, subpacked, nitrogen-filled, sealed, and sterilized at 121 ° C for 15 min to obtain a diclofenac fat emulsion injection.

The drug loading was determined to be 19.5 mg/ml; the average particle diameter was 270.1 nm; and the pH was 6.61. In accordance with the quality requirements of the present invention, it can be directly administered intravenously when used clinically.

Example 21: Example of a pharmaceutical preparation of the present invention

Weigh 1.5 g of diclofenac, 1.2 g of refined egg yolk lecithin, 5 g of soybean oil for injection, and 5 g of caprylic acid triglyceride for injection, and heat it to 74 ° C in a water bath, stir or shear to dissolve, and obtain oil. phase. The glycerin was dispersed in an appropriate amount of water for injection, mixed, and heated to 74 ° C to obtain an aqueous phase. The oil phase was slowly injected into the aqueous phase at 74 ° C while maintaining high shear for 15 min to obtain colostrum. This mixed emulsion was passed through a high pressure homogenizer, emulsified 3 times under a pressure of 13,000 psi, made up to 100 ml with water for injection, and adjusted to a pH of 6.03 with citric acid. It was filtered with a 0.22 μm pore size capsule filter, subpacked, nitrogen-filled, sealed, and sterilized at 121 ° C for 15 min to obtain a diclofenac fat emulsion injection.

The drug loading was determined to be 15 mg/ml; the average particle diameter was 181.7 nm; and the pH was 5.60. In accordance with the quality requirements of the present invention, it can be directly administered intravenously when used clinically.

Example 22: Example of a pharmaceutical preparation of the present invention

1 g of diclofenac ester, 10 g of soybean oil for injection, and 1.2 g of refined egg yolk lecithin were weighed, mixed, and heated to 74 ° C in a water bath, and stirred or sheared to dissolve to obtain an oil phase. The glycerin was dispersed in an appropriate amount of water for injection and heated to 74 ° C to obtain an aqueous phase. The oil phase was slowly injected into the aqueous phase at 74 ° C while maintaining high shear for 15 min to obtain colostrum. The mixed emulsion was passed through a high pressure homogenizer, emulsified 3 times under a pressure of 15,000 psi, made up to 100 ml with water for injection, and adjusted to a pH of 6.05 with phosphoric acid and disodium hydrogen phosphate. It was filtered with a 0.45 μm pore size capsule filter, subpacked, nitrogen-filled, sealed, and sterilized at 121 ° C for 15 min to obtain a diclofenac fat emulsion injection.

The drug loading was determined to be 10 mg/ml; the average particle diameter was 170.3 nm; and the pH was 5.45. In accordance with the quality requirements of the present invention, it can be directly administered intravenously when used clinically.

Example 23: Example of a pharmaceutical preparation of the present invention

1 g of diclofenac ester, 10 g of soybean oil for injection, and 1.2 g of refined egg yolk lecithin were weighed, mixed, and heated to 78 ° C in a water bath, and stirred or sheared to dissolve to obtain an oil phase. The glycerin was dispersed in an appropriate amount of water for injection and heated to 78 ° C to obtain an aqueous phase. The oil phase was slowly injected into the aqueous phase at 78 ° C while maintaining high shear for 10 min to obtain colostrum. This mixed emulsion was passed through a high pressure homogenizer, emulsified 3 times under a pressure of 12,000 psi, made up to 100 ml with water for injection, and adjusted to a pH of 6.02 with acetic acid. It was filtered with a 0.22 μm pore size capsule filter, subpacked, nitrogen-filled, sealed, and sterilized at 121 ° C for 15 min to obtain a diclofenac fat emulsion injection.

The drug loading was determined to be 10 mg/ml; the average particle diameter was 203.1 nm; and the pH was 5.57. In accordance with the quality requirements of the present invention, it can be directly administered intravenously when used clinically.

Example 24: Observing the analgesic effect of drugs by acetic acid writhing method (drug comparison)

According to the literature (Cui Yue. Diclofenac sodium lipid microsphere injection [D]. Shenyang Pharmaceutical University, 2009.) Preparation of 2mg / ml diclofenac sodium fat emulsion injection. Diclofenac fatty acid injection was prepared as in Example 16.

A total of 30 ICR mice, 6 in each group, were divided into blank group (normal saline), diclofenac sodium fat emulsion injection group (15mg/kg), diclofenac fat emulsion injection low dose group (9mg/kg), medium The dose group (18 mg/kg, the dose is equivalent to 15 mg/kg diclofenac sodium) and the high dose group (36 mg/kg). Each group of males and females was given the corresponding drugs. After 30 minutes, 0.2 mL of freshly placed 0.7% acetic acid saline solution was intraperitoneally injected, and the number of writhing in 30 min was recorded. The writhing phenomenon was: the abdomen contracted into an "S" shape. The torso and hind legs are stretched, the buttocks are raised and creeping. The results are shown in Table 2.

Table 2 Effect of diclofenac sodium fat emulsion injection and diclofenacate fat emulsion injection on the number of writhing in mice

Group Number of writhing Pain inhibition rate (%) Blank group 41.33±7.39 0 Diclofenac sodium fat emulsion (15mg/kg) 19.83±3.31* 62.63 Diclofenac fatty milk (9mg/kg) 24.67±8.82* 57.23 Diclofenac fatty milk (18mg/kg) 11.67±3.56*# 73.07 Diclofenac fatty milk (36mg/kg) 7.17±1.47*## 83.49

Compared with the blank control group: *P<0.001; compared with the diclofenac sodium group: #P<0.05; compared with the diclofenac sodium group##P<0.001.

Example 25: Observing the analgesic effect of a drug by acetic acid writhing method (Comparison of preparations)

A total of 30 ICR mice, 6 in each group, were divided into blank group (normal saline), diclofenac sodium injection group (15 mg / kg), diclofenac fat emulsion injection low dose group (9 mg / kg), medium dose group (18 mg/kg, equivalent to 15 mg/kg diclofenac sodium), high dose group (36 mg/kg). Each group of males and females was given the corresponding drugs. After 30 minutes, 0.2 mL of freshly placed 0.7% acetic acid saline solution was intraperitoneally injected, and the number of writhing in 30 min was recorded. The writhing phenomenon was: the abdomen contracted into an "S" shape. The torso and hind legs are stretched, the buttocks are raised and creeping. The results are shown in Table 3.

Table 3 Effect of diclofenac sodium solution and diclofenacate fat emulsion injection on the number of writhing in mice

Group Number of writhing Pain inhibition rate (%) Blank group 49.50±9.65 0 Diclofenac sodium solution (15mg/kg) 18.50±4.18* 62.63 Diclofenac ester emulsion (9mg/kg) 21.17±5.85* 57.23 Diclofenacate emulsion (18mg/kg) 13.33±2.73*# 73.07 Diclofenacate emulsion (36mg/kg) 8.17±1.47*## 83.49

Compared with the blank control group: *P<0.001; compared with the solution group: #P<0.05; compared with the solution group##P<0.001.

Conclusion: Diclofenac fat emulsion injection has obvious analgesic effect. At the same dose, the analgesic ability of diclofenac fat emulsion injection is stronger than that of diclofenac sodium solution, and there is significant difference (P<0.05). This experiment shows that diclofenac fat emulsion injection has a strong analgesic effect.

Conclusion: Diclofenac fat emulsion injection has obvious analgesic effect. At the same dose, the analgesic ability of diclofenac fat emulsion injection is stronger than that of diclofenac sodium fat emulsion injection, and there is significant difference (P<0.05). this experiment It shows that the analgesic effect of diclofenac is better than diclofenac sodium.

Example 26: Carrageenan-induced inflammation to observe the anti-inflammatory effects of drugs (drug comparison)

According to the literature (Cui Yue. Diclofenac sodium lipid microsphere injection [D]. Shenyang Pharmaceutical University, 2009.) Preparation of 2mg / ml diclofenac sodium fat emulsion injection and diclofenac ester fat emulsion injection.

Rats weighing 140-160 g, male and female, were randomly divided into groups of 6 each. Divided into blank group (normal saline), diclofenac sodium fat emulsion injection group (10mg/kg), diclofenac fat emulsion injection low dose group (6mg/kg), medium dose group (12mg/kg, dose equivalent to 10mg/ Kg diclofenac sodium), high dose group (24mg/kg). 0.1 ml of 1% carrageenan physiological saline solution was injected subcutaneously into the right hind toe of each rat to cause inflammation. After half an hour of inflammation, the tail veins were administered according to the above groups, 0.5, 1, after administration. The volume of the left and right hind legs was measured by volumetric method at 2, 3, 4, and 5 hours. The volume difference between the left and right hind paws before and after inflammation was the degree of swelling of the foot, and the swelling rate was calculated. The results are shown in Table 4.

Table 4 Diclofenac sodium fat emulsion injection, diclofenac ester fat emulsion injection on rat paw swelling rate after carrageenan inflammation

Compared with the blank group: *P<0.001, compared with the diclofenac sodium fat emulsion group #P<0.01;##P<0.001

Conclusion: Diclofenac sodium fat emulsion injection and diclofenac ester fat emulsion injection have a fast onset and have a rapid and strong anti-inflammatory effect at the beginning, while the same dose of diclofenac ester fat emulsion injection liquid is compared with diclofenac sodium fat emulsion. For injections, it shows a stronger, longer anti-inflammatory time. The experimental results show that the anti-inflammatory effect of diclofenac is stronger than that of diclofenac sodium.

Example 27: Carrageenan-induced inflammation to observe the anti-inflammatory effects of drugs (Comparison of preparations)

Rats weighing 140-160 g, male and female, were randomly divided into groups of 6 each. Divided into blank group (normal saline), diclofenac sodium injection group (10mg/kg), diclofenac fat emulsion injection low dose group (6mg/kg), medium dose group (12mg/kg, equivalent to 10mg/kg diclofenac sodium Solution), high dose group (24 mg/kg). 0.1 ml of 1% carrageenan physiological saline solution was injected subcutaneously into the right hind toe of each rat to cause inflammation. After half an hour of inflammation, the tail veins were administered according to the above groups, 0.5, 1, after administration. The volume of the left and right hind legs was measured by volumetric method at 2, 3, 4, and 5 hours. The volume difference between the left and right hind paws before and after inflammation was the degree of swelling of the foot, and the swelling rate was calculated. The results are shown in Table 5.

Table 5 Diclofenac sodium injection, diclofenac fat emulsion injection on rat paw swelling rate after carrageenan inflammation

Compared with blank group: *P<0.001, compared with solution group #P<0.01;##P<0.001

Conclusion: The results show that diclofenac sodium injection and diclofenac fat emulsion injection have a faster onset and have a rapid and strong anti-inflammatory effect at the beginning, while the same dose of diclofenac fat emulsion injection shows stronger anti-inflammatory. Effect, longer anti-inflammatory time.

Example 28: Hemolytic study of diclofenac fat emulsion injection

Take 10 ml of fresh rabbit blood, shake it in a glass flask-containing Erlenmeyer flask for 10 minutes, add 100 ml of 0.9% sodium chloride solution, shake well, centrifuge at 1000 r/min for 15 minutes, remove the supernatant, and precipitate the red blood cells again with 0.9. The % sodium chloride solution was washed 3 times as described above until the supernatant did not appear red. The resulting red blood cells were mixed with a 0.9% sodium chloride solution to form a 2% suspension for testing. Take 7 clean tubes and number them. Tubes 1 to 5 are for the test tube, tube No. 6 is the negative control tube, and tube No. 7 is the positive control tube. Add 2% red blood cell suspension, 0.9% sodium chloride solution or distilled water, diclofenac ester fat emulsion injection prepared in Example 15 as shown in Table 6. After mixing, immediately incubate in an incubator at 37 ° C. Observe whether there is red blood cell flocculation or hemolysis in each test tube, and observe once every 15 minutes. After 1 hour, observe once every 1 hour for a total of 3 hours. The experimental effect diagram after 3 hours is shown in Figure 3.

Table 6 diclofenac fat fat emulsion injection hemolysis test sample volume

Test tube number 1 2 3 4 5 6 7 2% red blood cell suspension (ml) 2.5 2.5 2.5 2.5 2.5 2.5 2.5 Saline (ml) 2.0 2.1 2.2 2.3 2.4 2.5 0 Distilled water (ml) 0 0 0 0 0 0 2.5 Diclofenac fat emulsion injection (ml) 0.5 0.4 0.3 0.2 0.1 0 0

According to Fig. 3, the solution in the No. 7 test tube was clear red, indicating that the positive control tube had hemolysis, the other test tube red blood cells were all sunk, and the supernatant liquid was colorless and clear, indicating that no hemolysis occurred. Experiments have shown that diclofenac fatty acid injection has no hemolysis.

Example 29: Diclofenac sodium injection and diclofenac fat emulsion injection vascular irritation test

Six rabbits weighing 2.2-2.5 kg, male and female, were randomly divided into two groups, A and B. Diclofenac sodium injection was diluted to a certain concentration with 0.9% physiological saline, and then dosed at a dose of 5.2 mg/kg in Group A. The left ear vein of the rabbit was administered by injection, and the administration volume was 2.1 ml, and 2.1 ml of 0.9% physiological saline was intravenously injected into the right ear margin. Each rabbit in group B was diluted with a dose of 6.2 mg/kg (equivalent to 5.2 mg/kg diclofenac sodium) in 0.9% saline to a certain concentration in the left ear vein. The diclofenac ester fat emulsion injection prepared according to Example 15 was 2.1 ml. At the same time, 2.1 ml of blank fat emulsion was injected intravenously into the right ear margin. Each rabbit in each group was given a time of 90 seconds. Twenty-four hours after the administration, the rabbits were placed at a distance of about 0.5 cm from the bilateral injection site of the rabbits. The specimens were placed in a 10% paraformaldehyde solution, and then the samples were taken, embedded in paraffin, and stained with HE. Histopathological examination was performed under light microscope.

Figure 4 shows partial tissue section images. The results show that the right ear veins of the rabbits in group A are intact, the vascular endothelium is smooth, no thrombus is seen in the uterine cavity, no bleeding, edema and inflammatory cell infiltration in the perivascular tissue, and group A. The left pericardial vein of the rabbit had edema and inflammatory cell infiltration, indicating that diclofenac sodium injection was severely irritating under the experimental conditions. In both groups, the rabbit ear veins were structurally intact, the vascular endothelium was smooth, no thrombus was seen in the uterine cavity, no bleeding, edema and inflammatory cell infiltration were observed in the perivascular tissue, indicating that diclofenacate fat emulsion injection was not seen in the blood vessels under the experimental conditions. Significant damage and irritation.

The preferred embodiments of the present invention have been specifically described above, but the present invention is not limited to the embodiments, and those skilled in the art can make various equivalents without departing from the inventive spirit of the present invention. Variations or substitutions are intended to be included within the scope of the invention as defined by the appended claims.

Claims (18)

A 1-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester having the following structural formula:

The method for synthesizing 1-(acetoxy)ethyl 2-[(2,6-dichlorophenyl)amino]phenylacetate according to claim 1, wherein the synthetic route of the method is:

The diclofenac sodium is reacted with 1-bromoethyl acetate in the presence of a basic catalyst to synthesize the compound of the invention. The method for synthesizing 1-(acetoxy)ethyl 2-[(2,6-dichlorophenyl)amino]phenylacetate according to claim 2, wherein the alkaline catalyst is One or more selected from the group consisting of potassium hydrogencarbonate, sodium hydrogencarbonate, sodium carbonate, and potassium carbonate. A method for synthesizing 1-[acetic acid]ethyl 2-[(2,6-dichlorophenyl)amino]phenylacetate according to claim 2 or 3, characterized in that the specific steps of the synthesis method as follows: A, adding 1 mol of sodium 2-[(2,6-dichlorophenyl)amino]phenylacetate to the reaction vessel, dissolving in an organic solvent, adding a basic catalyst, and stirring at room temperature; B, 1.1 to 2.0 mol of 1-bromoethyl acetate is added dropwise to the reaction solution, and reacted at room temperature for 5 to 30 minutes, and then heated to 30 ° C to 80 ° C to carry out a reaction; C. After the reaction was stopped, the organic solvent and water were added to the reaction mixture, and the organic phase was separated, washed with water, washed with a sodium thiosulfate aqueous solution, and finally washed with saturated brine. D. The organic phase is rotated to evaporate the organic solvent, and the crude product is dried under vacuum to obtain. The method for synthesizing 1-(acetoxy)ethyl 2-[(2,6-dichlorophenyl)amino]phenylacetate according to claim 4, wherein the organic solvent is N, One or more of N-dimethylacetamide, N,N-dimethylformamide, and acetone; and the amount added is 0.5 to 1 L. The method for synthesizing 1-(acetoxy)ethyl 2-[(2,6-dichlorophenyl)amino]phenylacetate according to claim 4, wherein the basic catalyst is added The amount is 0.1 to 1.2 mol, and the mixture is stirred at room temperature for 0.5 to 4 hours. The method for synthesizing 1-(acetoxy)ethyl 2-[(2,6-dichlorophenyl)amino]phenylacetate according to claim 4, wherein the organic phase is separated by extraction, and acetic acid is selected. One or more of ethyl ester, diethyl ether and methyl tert-butyl ether are used as extracting agents, and the total amount of solvent added is 0.5-5 L, wherein the volume ratio of extractant to water is 1:1 to 1.5. The pH of the aqueous sodium thiosulfate solution is 9-11. A fat emulsion injection containing 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to claim 1, characterized in that the fat emulsion The injection is composed of diclofenac ester, injectable oil, refined lecithin, glycerin, pH adjuster and water for injection. The percentage of each component relative to the total volume of the injection is as follows:

The injection oil is a mixture of soybean oil for injection or soybean oil for injection and caprylic acid triglyceride; and the purified lecithin is egg yolk lecithin. A fat emulsion injection of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to claim 8, wherein the diclofenac salt is 0.8 to 1.5% g/ml. A fat emulsion injection of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to claim 8, wherein the diclofenac salt is 1% g/ml. The fat emulsion injection of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to claim 8, wherein the injection oil The dosage is 10% g/ml. The fat emulsion injection of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to claim 8, wherein the injection oil A mixture selected from the group consisting of soybean oil for injection, or soybean oil for injection and caprylic acid triglyceride for injection, wherein the soybean oil for injection is contained in the mixture in an amount of not less than 50% by weight. The fat emulsion injection of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to claim 8, wherein the pH adjuster It is selected from one or two of citric acid, hydrochloric acid, phosphoric acid, acetic acid, disodium hydrogen phosphate, and sodium hydroxide. The fat emulsion injection of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to claim 8, wherein the pH adjuster The pH was adjusted to 6.5 to 7.5. A fat emulsion injection of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to any one of claims 8 to 14, wherein said The average thickness of the fat emulsion is 170 to 300 nm. A method for preparing a fat emulsion injection of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to any one of claims 8 to 14, which is characterized in that The preparation method is: Weigh an appropriate amount of oil for injection, refined egg yolk lecithin, mix, and heat to 55 ° C ~ 80 ° C in a water bath, stir or shear to dissolve; then add diclofenac ester at 55 ° C ~ 80 ° C stir or shear to dissolve To produce an oil phase; The oil is dispersed in the prescribed amount of water for injection, heated to 55 ° C to 80 ° C to obtain an aqueous phase; the oil phase is slowly injected into the aqueous phase at 55 ° C to 80 ° C while maintaining high shear for 10 to 15 minutes. Milk; colostrum is further emulsified by a high-pressure homogenizer at a homogenization pressure of 10,000 to 15,000 psi, pH is adjusted with a pH adjuster, filtered, dispensed, nitrogen-filled, sealed, and sterilized. The method for preparing a fat emulsion injection of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to any one of claims 16, wherein The lecithin is dissolved in the oil phase or dispersed in the aqueous phase. The method for preparing a fat emulsion injection of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid 1-(acetoxy)ethyl ester according to any one of claims 16, wherein The filtration and sterilization described are respectively capsule filtration and autoclaving, wherein the capsule filter has a filtration pore size of 0.45 μm or 0.8 μm, and the autoclave sterilization temperature is 121 ° C for 15 minutes.

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