WO2017143877A1 - 供注射用双氯芬酸钠药物组合物及其制备方法 - Google Patents

供注射用双氯芬酸钠药物组合物及其制备方法 Download PDF

Info

Publication number
WO2017143877A1
WO2017143877A1 PCT/CN2017/070685 CN2017070685W WO2017143877A1 WO 2017143877 A1 WO2017143877 A1 WO 2017143877A1 CN 2017070685 W CN2017070685 W CN 2017070685W WO 2017143877 A1 WO2017143877 A1 WO 2017143877A1
Authority
WO
WIPO (PCT)
Prior art keywords
injection
diclofenac sodium
solution
pharmaceutical composition
preparation
Prior art date
Application number
PCT/CN2017/070685
Other languages
English (en)
French (fr)
Inventor
吴夙慧
张涛
陈锡平
Original Assignee
江苏思邈医药科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 江苏思邈医药科技有限公司 filed Critical 江苏思邈医药科技有限公司
Publication of WO2017143877A1 publication Critical patent/WO2017143877A1/zh

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/196—Carboxylic acids, e.g. valproic acid having an amino group the amino group being directly attached to a ring, e.g. anthranilic acid, mefenamic acid, diclofenac, chlorambucil
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions

Definitions

  • the invention belongs to the technical field of medicine, and in particular relates to a pharmaceutical composition for diclofenac sodium for injection and a preparation method thereof.
  • Diclofenac Sodium is a phenylacetic acid derivative. It is a third-generation potent non-steroidal anti-inflammatory analgesic. It is also known as diclofenac, tamarind, and Amirel. It is a white crystalline powder and odorless. Soluble in acetone, soluble in methanol and ethanol, slightly soluble in water, hygroscopic, its molecular formula is C 14 H 10 Cl 2 NNaO 2 , molecular weight is 318.
  • the structural formula of diclofenac sodium is as follows:
  • diclofenac sodium The main mechanism of action of diclofenac sodium is to inhibit cyclooxygenase activity, thereby blocking the conversion of arachidonic acid to prostaglandins. At the same time, it can also promote the combination of arachidonic acid and triglyceride, reduce the concentration of intracellular free arachidonic acid and indirectly inhibit the synthesis of leukotrienes, thereby achieving the purpose of analgesia and anti-inflammatory. Its clinical antipyretic and analgesic effect is exact, widely used in rheumatoid rheumatoid, lumbar disc herniation, cervical vertebra pain, toothache and other fields.
  • the dosage forms of commercially available diclofenac sodium are mainly oral dosage forms such as tablets and capsules, topical drugs such as rubbing agents, suppositories, gels, and injections, and lyophilized preparations for injection of diclofenac sodium have not been listed.
  • the above-mentioned commercially available diclofenac sodium preparations have obvious deficiencies: oral and topical dosage forms are not ideal in terms of bioavailability and onset rate, and administration often causes more severe gastrointestinal reactions and first-pass effects.
  • Diclofenac sodium injection can overcome the above-mentioned shortcomings, but due to the low solubility of diclofenac sodium in water (generally at 9 mg/mL, 25 ° C), the formulation concentration requirement is not reached, and the structure of diclofenac sodium contains a group susceptible to oxidation. The stability of the drug is reduced after it is prepared into an injection.
  • Diclofenac sodium injection (Voltarol, 75mg/3mL) developed by Novartis Pharmaceuticals is administered by intramuscular injection or intravenous drip with propylene glycol and benzyl alcohol as a solubilizer. It is highly irritating to blood vessels. It has strong pain when administered. It may cause venous inflammation in long-term application.
  • Diclofenac sodium cyclodextrin inclusion injection (Dyloject, 75mg/2mL) was developed by Shimoda Biotech and approved by Javelin Pharmaceuticals using cyclodextrin technology. It was marketed in the UK in December 2007 for the treatment of acute pain. It can be administered by intramuscular or intravenous injection.
  • the dosage form encapsulates diclofenac sodium with hydroxypropyl- ⁇ -cyclodextrin, which effectively solves the problem of poor solubility of the main component, and uses thioglycerol as an antioxidant to prevent oxidative degradation of the drug.
  • diclofenac sodium with hydroxypropyl- ⁇ -cyclodextrin, which effectively solves the problem of poor solubility of the main component, and uses thioglycerol as an antioxidant to prevent oxidative degradation of the drug.
  • the preparation was temporarily recalled due to the discovery of unknown white particles in some batches of preparation bottles, which temporarily withdrew from the UK market. This was incompatible with the long-term stability of the inclusion of diclofenac sodium in the cyclodextrin aqueous solution. Drug leakage is related to the precipitation of crystals.
  • the administration method was intravenous administration.
  • the formulation was the same as the 75mg/2mL specification, using hydroxypropyl- ⁇ .
  • -Cyclodextrin improves drug solubility (333mg/mL)
  • injection time needs to be greater than 15s also has long-term stability problems, in its FDA instructions, dosage and usage It is clearly stated that it should be observed whether there are visible particles and discoloration before use. If there are particles or discoloration, it cannot be used, indicating that there is a risk of drug crystallization and oxidative degradation.
  • Patent CN 94118042 discloses an injectable pharmaceutical composition or veterinary composition comprising diclofenac or a salt thereof and 2-hydroxypropyl- ⁇ -cyclodextrin, which is prepared by acidifying an aqueous solution without pH addition.
  • the buffer the lower limit of the pH of the composition is acidic, and the long-term stability of the composition is not studied; the highest concentration of diclofenac sodium in the composition is only 25 mg/ml, and the solution also contains sodium metabisulfite and chloroprene.
  • a preservative such as alcohol.
  • Chinese patent CN 1711996A discloses an injectable pharmaceutical composition containing diclofenac sodium and ⁇ -cyclodextrin in the form of an aqueous solution containing diclofenac sodium, ⁇ -cyclodextrin and polysorbate, which is suitable only for subcutaneous And muscle administration.
  • Chinese patent CN 200410024064 discloses a lyophilized preparation containing diclofenac salt and lidocaine, and a preparation method thereof, which works by two pharmacologically active components of diclofenac and lidocaine, which employs a surfactant to spit As a solubilizer, Wen 80 is added with a preservative in the prescription, which has great irritating and toxic side effects on the patient, and does not consider the oxidative degradation of the drug.
  • Patent CN 101244278A discloses a preparation for intravenous injection of diclofenac sodium and a preparation method thereof, which comprises using hydroxypropyl- ⁇ -cyclodextrin to encapsulate diclofenac sodium, and adding an antioxidant to the preparation, Considering the stability and toxic side effects of the formulation, the formulation showed no significant improvement compared to the commercially available Dyloject.
  • Patent CN 101123957A discloses an injection of diclofenac and a pharmaceutically acceptable salt thereof, which is in the form of a solution containing a large amount of additives such as an alcohol cosolvent, an antioxidant, a preservative, etc., which has a large side effect and can only be used for Intramuscular injection and slow intravenous bolus.
  • the present invention adopts a specific pharmaceutical composition and preparation method of diclofenac sodium, thereby obtaining stable, safe and reliable quality.
  • a pharmaceutical composition for diclofenac sodium for injection having a small toxic side effect.
  • the medicine composition for diclofenac sodium for injection can be directly injected intravenously, and can be used for treating mild, moderate and severe pain, wherein the amount of cyclodextrin derivative is low, and a small amount of solvent is re-dissolved and isotonic with human blood. Can significantly reduce the pain of the injection site, speed up the drug injection time (5 ⁇ 10s), improve the analgesic effect;
  • the prescription does not need to add antioxidants, but also avoid the risk of precipitation of crystals during the placement process, significant It increases the stability of the preparation, improves the safety of clinical application, increases the compliance of the patient's administration, and creates a higher clinical therapeutic benefit.
  • a pharmaceutical composition for diclofenac sodium for injection which is stable, effective and safe, and which comprises diclofenac sodium, a water-soluble cyclodextrin derivative, and a stabilizer.
  • the pharmaceutical composition for diclofenac sodium for injection of the present invention is a lyophilized powder injection prepared by a freeze-drying process of a solution prepared from a formulation containing diclofenac sodium, a water-soluble cyclodextrin derivative, a stabilizer, and water for injection.
  • the solution contains or preferably consists of the following components:
  • the residual oxygen amount is controlled to be less than 2%, and the dissolved oxygen amount is controlled to be less than 1 mg/L.
  • the formulation contains or preferably consists of the following components:
  • the water-soluble cyclodextrin derivative may be injectable from at least one selected from the group consisting of hydroxypropyl- ⁇ -cyclodextrin, sulfobutyl- ⁇ -cyclodextrin, and glucosyl- ⁇ -cyclodextrin.
  • the water-soluble cyclodextrin derivative used is preferably hydroxypropyl- ⁇ -cyclodextrin.
  • the present invention relates to the inclusion of diclofenac sodium by the inclusion of a water-soluble cyclodextrin derivative to increase the solubility of diclofenac sodium, wherein the inventors of the present invention have found that diclofenac sodium and a water-soluble cyclodextrin derivative are dissolved in water. After that, in the solution of diclofenac sodium concentration of 37.5 mg/mL, maintaining the pH range of the system is 7-10, which can significantly increase the inclusion degree of diclofenac sodium, which may be because the alkaline environment can affect the three-dimensional structure of the inclusion complex. Increase the number of voids provided by the cyclodextrin to promote the inclusion of diclofenac sodium.
  • the pH of the solution is selected to be in the range of 7.0-10.0, preferably pH 8.0-9.0.
  • the stabilizer is a pharmaceutically acceptable injectable salt, for example, various buffer salt systems (such as tromethamine, phosphate (such as disodium hydrogen phosphate), citric acid-sodium citrate, etc.), or
  • the buffer salt system is combined with sodium hydroxide, hydrochloric acid or the like.
  • the stabilizer is a tromethamine or phosphate buffer system, or a combination thereof with sodium hydroxide.
  • the inclusion of the water-soluble cyclodextrin derivative and diclofenac sodium is in a dynamic equilibrium, which is affected by a variety of factors.
  • the inventors of the present invention have found that the use of the stabilizer having a buffering ability can reduce or avoid the crystallization phenomenon after re-dissolution of the obtained lyophilized preparation, and remarkably improve the stability of the clathrate.
  • the occurrence of crystallization of the inclusion complex may be related to the decrease of the pH value during the solution placement of the lyophilized preparation after re-dissolution, thereby destroying the dynamic balance of the inclusion complex.
  • the water-soluble cyclodextrin derivative cannot be completely contained after the dynamic equilibrium is destroyed.
  • Diclofenac sodium causes poor stability of the package and, in turn, causes crystallization of the drug.
  • the stabilizer according to the present invention to control the pH range of the prescription, the pH value of the system can be prevented from being significantly changed, the dynamic equilibrium state of the clathrate can be maintained, the crystallization phenomenon can be reduced or avoided, and the stability of the clathrate can be improved, and the salt is also buffered. It can be combined with water molecules remaining in the lyophilized preparation to avoid competitive binding of water molecules to diclofenac sodium to cyclodextrin derivative van der Waals (Vander Waals) force action site.
  • Vander Waals van der Waals
  • Another object of the present invention is to provide a method for preparing a pharmaceutical composition for diclofenac sodium for injection which is convenient for industrial mass production and simple in process, and comprises the following steps:
  • the bottle is filled with an inert gas, the residual oxygen amount is controlled to be less than 2%, the dissolved oxygen amount is controlled to be less than 1 mg/L, and the filling is performed according to the preparation unit. (packing) into each of the bottles, half-plugged;
  • the drug solution obtained in the step 4) is freeze-dried, tamped, glanded under aseptic conditions, and sealed to obtain a diclofenac sodium drug composition.
  • the stirring time is preferably 1-3 h, thereby completing the inclusion of the water-soluble cyclodextrin derivative on diclofenac sodium, and improving the solubility of diclofenac sodium.
  • the stabilizer is such that when the pharmaceutical composition for diclofenac sodium for injection is reconstituted with water to a solution having a diclofenac sodium concentration of 37.5 mg/mL, the stabilizer can control the pH range of the solution to 7-10, preferably 8.0-9.0.
  • the stabilizer is a pharmaceutically acceptable injectable salt, for example, various buffer salt systems (such as tromethamine, phosphate (such as disodium hydrogen phosphate), citric acid - Sodium citrate or the like, or a combination of the buffer salt system and sodium hydroxide, hydrochloric acid or the like.
  • the stabilizing agent is a tromethamine or phosphate buffer system, or a combination thereof with sodium hydroxide.
  • the alkaline storage stabilizer having a buffering ability is advantageous for ensuring stable incorporation of the drug and ensuring product stability.
  • the amount of the activated carbon is preferably 0.02%-0.1% of the total weight of the prescription solution in the step 1), and the amount of the activated carbon is too small, the solution may not be completely decolored, depyrogenated and decontaminated, and the amount is too much.
  • the active ingredient in the solution is adsorbed, so the amount of activated carbon should be controlled within a reasonable range.
  • the inert gas is used to control the residual oxygen amount and the dissolved oxygen amount, to protect the diclofenac from oxidative degradation and to improve the stability of the product, and the inert gas is preferably nitrogen.
  • the freeze-drying process is a conventional freeze-drying process in the art.
  • the freeze-drying process may preferably: put the bottle that has been dispensed into the freeze-drying box, and cool down to -40. °C, pre-freezing at -40 °C for 1-3h, then vacuum; temperature to -29 ° C, for 3-5h; warm to -10 ° C, for 4-6h; warm to 0 ° C, for 1h; °C, keep 5-8h.
  • the invention is not limited to this.
  • the vacuum plug or the inert gas filled with a certain pressure may be used, and the embossing of the above two conditions makes the bottle have no oxygen, which can prevent the obtained product from being Oxidative degradation reactions occur during the placement process.
  • the introduction of oxygen will affect the stability of this product.
  • the long-term and accelerated June stability studies showed that the product stability was good, the oxidative degradation impurities did not increase, and the appearance of the composition did not undergo a color change caused by the oxidation reaction.
  • the content of the active drug diclofenac sodium in the pharmaceutical composition for diclofenac sodium for injection is 75 mg per bottle or 37.5 mg per bottle.
  • the pH ranges from 7.0 to 10.0, preferably from 8.0 to 9.0.
  • Another object of the present invention is to provide a pharmaceutical composition for diclofenac sodium for injection which is stable, effective and safe, and which is a lyophilized powder injection prepared by the above method.
  • Another object of the present invention is to provide a solution for preparing the pharmaceutical composition for diclofenac sodium for injection, wherein the solution comprises the following components: diclofenac sodium; water-soluble cyclodextrin derivative; stabilizer ; and water for injection.
  • Another object of the present invention is to provide the use of the pharmaceutical composition of diclofenac sodium for injection for the treatment of mild, moderate and severe pain.
  • the dosage of cyclodextrin is significantly lower than the existing commercial Dyloject, thereby reducing the side effects such as nephrotoxicity and hemolysis; the antioxidant is not needed in the prescription solution, which reduces the safety hazard.
  • the types and amounts of excipients in the preparations are small, which reduces the damage to the human body, and its safety meets the requirements of relevant national laws and regulations;
  • good stability, freeze-dried powder needle preparation form can avoid the risk of precipitation of crystal precipitation during long-term placement of the injection; use a special buffer system to control the pH value of the preparation, enhance the water-soluble cyclodextrin derivative on diclofenac sodium In combination with the control; the amount of residual oxygen is controlled, and the possibility of oxidation reaction is low; the impurity in the lyophilized preparation is less than 0.2%, which significantly increases the stability of the preparation and makes the quality reliable;
  • efficacy and patient compliance, reconstituted injection has the same medical properties as Dyloject, and compared to Dyloject injection time greater than 15s, can further accelerate the injection time (5 ⁇ 10s), significantly faster
  • the onset time of the drug can be used to treat mild, moderate, and severe pain, to quickly reduce the pain of the patient, and to reduce the pain discomfort at the injection site and increase the compliance of the patient;
  • the invention has low cost, and the pharmaceutical composition prepared by the invention has few kinds of auxiliary materials and low dosage, and the obtained lyophilized preparation can reach or exceed other drugs of the same type, which fully proves the outstanding advantages of the medicine of the invention;
  • the preparation method of the diclofenac sodium sodium pharmaceutical composition for injection according to the invention is simple, efficient, and low in cost, and is suitable for industrial application.
  • Fig. 1 is an X-ray diffraction chart showing a physical mixture of a simple mixing of a prescription (other than water for injection) of Preparation Example 1 according to the present invention.
  • FIG. 2 is an X-ray diffraction chart showing the pharmaceutical composition for diclofenac sodium for injection prepared according to Preparation Example 1 of the present invention for 0 months.
  • Figure 3 is a graph showing the X-ray diffraction pattern of the pharmaceutical composition for diclofenac sodium for injection prepared according to Preparation Example 1 of the present invention accelerated for 6 months.
  • Figure 4 is a graph showing the X-ray diffraction pattern of the pharmaceutical composition for diclofenac sodium for injection prepared according to Preparation Example 1 of the present invention for a long period of 24 months.
  • Figure 5 is a graph showing the drug time after intravenous administration of a diclofenac sodium pharmaceutical injection preparation prepared by the preparation example 4 of the present invention and the diclofenac sodium drug preparation prepared in Comparative Example 1.
  • Fig. 6 is a partial enlarged view of the first 2 hours in the medicine time chart shown in Fig. 5.
  • diclofenac sodium Anyang Kyushu Pharmaceutical Co., Ltd.
  • hydroxypropyl- ⁇ -cyclodextrin Roquette (China) Fine Chemical Co., Ltd.
  • sulfobutyl- ⁇ -cyclodextrin USA
  • Captisol sodium hydroxide
  • tromethamine Wang Chemical Co., Ltd.
  • disodium hydrogen phosphate Hunan Jiudian Pharmaceutical Co., Ltd.
  • water for injection homemade
  • Conventional unlabeled reagents were purchased from Sinopharm Chemical Reagent Co., Ltd.
  • the preparation method is:
  • the preparation method is:
  • the preparation method is:
  • the preparation method is:
  • the preparation method comprises the following steps: dissolving hydroxypropyl- ⁇ -cyclodextrin in a part (about 90%) of water for injection, adding diclofenac sodium, stirring until completely dissolved, adding sodium hydroxide to stir and dissolve, and adding the remaining amount (about 10%) of water for injection; then sterilized by filtration on a 0.22 ⁇ m filter, filled in a 1 mL/bottle, stoppered, capped, and sterilized at 116 ° C for 15 min to obtain diclofenac sodium injection.
  • the present inventors have found that the diclofenac sodium injection prepared by the commercially available Dyloject formulation produces a large amount of degradation material (from 0.05% to 1.27%) in the high temperature sterilization step, indicating that the product is sensitive to high temperatures.
  • the preparation method of the invention does not require the high temperature sterilization step of the injection solution, and can significantly reduce the impurity quality of the product and improve the stability.
  • the preparation method is:
  • the preparation method is:
  • the obtained chemical liquid is divided into the respective glass bottles according to the preparation unit, and the plug is half-filled;
  • a plurality of bottles of the pharmaceutical composition for diclofenac sodium for injection obtained in Preparation Example 1 were reconstituted by adding 1 ml of water per bottle to examine the reconstitution stability of the preparation.
  • the investigation time was 0h, 2h, 4h, 8h, 12h, 24h.
  • the indicators were clarity, alkalinity, content, related substances, visible foreign matter and insoluble particles. The measurement results are shown in Table 1.
  • test results show that the stability of the pharmaceutical composition for diclofenac sodium for injection of the present invention is good within 24 hours after reconstitution.
  • the diclofenac sodium injection prepared in Example 1 (same as the commercially available Dyloject prescription), the diclofenac sodium lyophilized preparation prepared in Comparative Example 2 and Comparative Example 3 was accelerated (40 ° C ⁇ 2 ° C / 75% RH ⁇ 5% RH) 6 months and long-term (25 ° C ⁇ 2 ° C / 60% RH ⁇ 5% RH) 24 months test study, key indicators for the sample: traits, clarity, alkalinity (reconstituted with water for injection) The solution containing diclofenac sodium (37.5 mg/ml), visible foreign matter, insoluble particles, related substances and contents were examined. The test results are shown in Table 2.
  • the pharmaceutical composition for diclofenac sodium for injection prepared by the present invention has quality assurance and good stability.
  • the physical mixture powder was subjected to X-ray diffraction measurement to investigate the inclusion of hydroxypropyl- ⁇ -cyclodextrin on the drug.
  • the detection conditions were 3.0 to 40.0°, 0.01°/second, acceleration voltage: 40KV, and current 40 mA. The test results are shown in Figures 1 to 4.
  • the pharmaceutical composition for diclofenac sodium for injection obtained by the preparation method of the present invention is subjected to a special safety test: a hemolytic test, an irritant test, and an allergic test.
  • a hemolytic test a hemolytic test
  • the diclofenac sodium injection prepared in Comparative Example 1 was used as a control preparation.
  • Preparation of 2% rabbit red blood cell suspension Take a test rabbit (animal: rabbit, source: Shanghai Animal Research Institute Experimental Animal Center), take about 20ml of blood in the ventricle under awake state, stir with cotton swab for about 10 minutes to remove fibrin Originally, pour into the test tube and add about 10 times the amount of 0.9% sodium chloride injection. After centrifugation at 1,500 rpm for 15 minutes, the supernatant was removed, and the precipitated red blood cells were washed by centrifugation with 0.9% sodium chloride injection three times or more until the supernatant was not red. The resulting red blood cells were mixed with a 0.9% sodium chloride injection into a 2% red blood cell suspension for testing.
  • red blood cell agglomeration occurs, it can be further determined whether it is true or false. If the agglomerates are evenly dispersed after shaking, or the agglomerates are placed on the glass slide, add 2 drops of 0.9% sodium chloride solution on the edge of the coverslip, and observe under the microscope, the condensed red blood cells can be disintegrated as false Condensation, on the contrary, the agglomerates are not shaken or are not smashed on the slides.
  • test drug can be used for injection; if the solution in the 1-5 tube is hemolyzed and agglomerated within 3 hours, the test drug is not available for injection.
  • the results of the hemolysis test were as follows: the No. 7 test tube was red-clear immediately after adding distilled water, and there was no red blood cell residue at the bottom of the tube, indicating that all hemolysis was observed. Red blood cells begin to sink at 15 min in test tube solution No. 1-6, with time delay When the long-term sinking red blood cells gradually increased, the supernatant liquid gradually increased, the supernatant was light yellow and clear, indicating that the test solution and the reference sodium chloride injection had no hemolysis, shaking the test tubes after 3 hours, 1-6 The red blood cells of the test tube can be evenly dispersed without red blood cell agglomeration. The specific test results are shown in Table 5.
  • Dose design The clinical recommended dose of diclofenac sodium is 37.5 mg/time, which is converted to rabbit dose of 1.9 mg/kg, so the rabbit test dose is determined to be 2 mg/kg.
  • mice Eight rabbits (animal: rabbit, source: Shanghai Animal Research Laboratory Experimental Animal Center) were divided into two groups: experimental group and control group, 4 groups/group, male and female, fixed in rabbit fixed box, left and right ears.
  • the hair was removed from the hair, the alcohol was disinfected, and the needle was injected from the farthest heart end of the ear vein.
  • the left and right ear veins of the test group were intravenously injected with 0.9% sodium chloride injection (control side) and the diclofenac sodium drug for injection according to the present invention.
  • the test solution (administration side) of the composition was intravenously injected at a dose of 2 mg/kg body weight at an injection rate of about 5 to 10 s; the left and right ear veins of the control group were intravenously injected with 0.9% sodium chloride injection ( The control side) and the diclofenac sodium control preparation (administration side) prepared in Comparative Example 1 were intravenously injected at a dose of 2 mg/kg body weight, and the injection speed was about 15 s to complete the injection.
  • Each group was given a needle once a day for 5 consecutive days.
  • the daily administration site was gradually advanced from the telecentric end to the proximal end of the ear vein.
  • the rabbit ear vein reaction was visually observed, including whether the blood vessel was clear, whether there was vasodilation or Contraction, whether there is congestion in the blood vessels, no edema around the blood vessels.
  • the experimental group of rabbits had clear veins of the ear veins, no vasodilation or contraction, no obvious congestion in the blood vessels, no edema around the blood vessels, and there was no venous vein in the control side. Significant improvement.
  • tissue biopsy showed that the vascular structures of the a and b segments of the rabbit ear were normal at 48 hours, 96 hours and 14 days after the last administration of the experimental group. There was no obvious vasodilatation and congestion, and no obvious thrombosis.
  • Vascular endothelial cells were basically normal, no swelling, necrosis and other phenomena, no inflammatory tissue around the blood vessels, no inflammatory cell infiltration, the average score was 0, no significant difference compared with the control side.
  • the vascular structures of the a and b segments of the rabbit ear were normal, some showed vasodilatation and congestion, no obvious thrombosis, and the vascular endothelial cells were basically normal. No swelling, necrosis and other phenomena, edema around the perivascular tissue, no inflammatory cell infiltration, the average score was 0.75, showing mild irritation.
  • the above test results showed that the diclofenac sodium prepared in the preparation example 4 was intravenously injected into the ear of the test solution at a rate of 37.5 mg/ml, once daily for 5 days, and no obvious local irritation reaction was observed in the rabbit ear vein.
  • the control preparation of Comparative Example 1 prepared according to the commercially available prescription was intravenously injected with 37.5 mg/ml of the ear vein once a day for 5 days, showing a mild irritation response to the rabbit ear vein. It can be seen that the diclofenac sodium pharmaceutical composition of the present invention has a significant improvement in irritation and is safe.
  • Dose design The clinical recommended dose of diclofenac sodium is 37.5 mg/time, which is converted to 2.9 mg/kg for guinea pigs. Therefore, the low dose of systemic active allergy test in guinea pigs is determined to be 3.0 mg/kg, and the high dose is determined to be 6.0 mg/kg.
  • guinea pigs (animals: guinea pigs, source: Shanghai Animal Research Institute Experimental Animal Center) were randomly divided into 4 groups according to their gender, 6 in each group, half male and half female.
  • the first group was used as a negative control group, and 0.9%/100 g of 0.9% sodium chloride injection was intraperitoneally injected once every other day for 3 times.
  • the second group was used as a positive control group, and 5% fresh egg white solution 0.2ml/100g was intraperitoneally injected once every other day for 3 times.
  • the third group was used as a low-dose group, and the low-dose solution of diclofenac sodium was intraperitoneally injected (1 ml of test solution of 37.5 mg/ml diclofenac sodium was placed in a 25 ml volumetric flask, and 0.9% sodium chloride injection was added to dissolve the volume) 0.2ml/100g, once every other day, for a total of 3 times.
  • the fourth group was used as a high-dose group, and the high-dose solution was prepared by intraperitoneal injection of diclofenac sodium.
  • 2 healthy sensitized guinea pigs can be taken from the intravenous dose of the drug to observe whether there is any similar allergic reaction caused by the test drug.
  • Antiserum preparation 8 guinea pigs were randomly divided into 4 groups according to their body weight, 2 in each group, half male and half female.
  • the first group served as a negative control group, intraperitoneal injection of 0.9% sodium chloride injection 0.2ml / 100g, separated Inject once a day for 5 times.
  • the second group was used as a positive control group, and intraperitoneal injection of 2 mg/kg egg protein solution 0.2 ml/100 g was given once every other day for 5 times.
  • the third group was used as a low-dose administration group, and the above-mentioned diclofenac sodium test preparation low-dose solution 0.2 ml/100 g was intraperitoneally injected once every other day for 5 times.
  • the fourth group was used as a high-dose administration group, and the above-mentioned diclofenac sodium test preparation high-dose solution 0.2 ml/100 g was intraperitoneally injected once every other day for 5 times.
  • the guinea pigs in each group were bled on the 14th day after the last sensitization, and naturally coagulated at room temperature.
  • the serum was separated by centrifugation at 2000 rpm for 20 min, and then stored in a refrigerator at -18 °C for use.
  • Passive skin allergy test 24 guinea pigs were randomly divided into 4 groups according to their body weight, 6 rats in each group, male and female, respectively, which were negative control group, positive control group, low dose group and high dose group.
  • the guinea pig's back skin was depilated with an animal epilator, and the above antiserum dilutions (1:2, 1:8, 1:32, v/v) were prepared with 0.9% sodium chloride injection, respectively.
  • the corresponding antiserum dilutions were injected intradermally into the back of the hair removal site, 0.1 ml per point, and the interval between the two points was about 2.5 cm.
  • Stimulation injection After 24 hours, the corresponding solutions of each group were prepared in the same manner as in the sensitization, and mixed with 1% Evans blue solution in a ratio of 1:1, and intravenously administered 0.4 ml/100 g. After 30 minutes of stimulating injection, the guinea pigs were sacrificed by bloodletting, the back skin was cut out, and the spot diameter of the inner layer of the skin was measured with a vernier caliper. The diameter of the irregular spots was half of the sum of the long diameter and the short diameter, and the diameter was greater than 5 mm, and the positive reaction was calculated. percentage.
  • the pharmaceutical composition for diclofenac sodium for injection prepared in Preparation Example 4 was reconstituted with 1 ml of water per bottle to prepare a test solution having a concentration of 37.5 mg/ml, and the analgesic effect of the drug was examined in a pain animal model. .
  • the diclofenac sodium injection prepared in Comparative Example 1 was used as a control solution. Each 1 ml of the test solution and the control solution were separately placed in a 50 ml volumetric flask, and the volume was diluted with physiological saline to a concentration of 0.75 mg/ml.
  • mice Take healthy Kunming mice (animal: mouse, source: Shanghai Institute of Drug Research Laboratory Animal Center) 30, weighing about 20g, male and female, randomly divided into 3 groups, according to 7.5mg / kg tail vein injection of mice Each group of mice was intraperitoneally injected with 0.2 mL of 0.6% glacial acetic acid solution 10 min after administration, and immediately counted. The number of writhing reactions of the mice within 15 min was observed, and the inhibition rate of the writhing reaction was calculated.
  • the specific grouping and administration methods are shown in Table 16, and the test results are shown in Table 17.
  • the pharmaceutical composition for diclofenac sodium for injection prepared in Preparation Example 4 was reconstituted with 1 ml of water per bottle to prepare a test solution having a concentration of 37.5 mg/ml, and the drug-generating power of the drug was examined by using a canine animal model. Learning behavior.
  • the diclofenac sodium injection prepared in Comparative Example 1 was used as a control solution.
  • the test animals should be adapted for breeding at the test site 3-7 days before the test day.
  • the administration solution was prepared by using the self-control cross-test design and the test solution prepared by reconstitution of water of Preparation Example 4 and the injection of Comparative Example 1, respectively.
  • the dose administered was 37.5 mg/head.
  • the interval between the two weeks is not less than 5 days.
  • the administration solution of Preparation Example 4 was completed in 5 to 10 s, and the administration solution of Comparative Example 1 was injected for more than 15 s.
  • the concentration of the drug in the plasma was determined by LC-MS. Chromatographic conditions: Agilent XDB C18 (150 mm ⁇ 4.6 mm, 5 ⁇ m) was used for the column, and the mobile phase was acetonitrile-water (containing 4 mM ammonium acetate, 0.08% formic acid) (75:25, V/V) at a flow rate of 0.8 mL ⁇ min. -1 , column temperature 25 ° C, injection volume is 5 ⁇ L.
  • Mass spectrometry conditions electrospray ionization source (ESI), gas curtain gas (CUR) pressure of 20 psi, spray gas (GS1) pressure of 55 psi, dry gas (GS2) pressure of 55 psi, source injection voltage (IS) of -4000 V, ion source
  • ESI electrospray ionization source
  • CUR gas curtain gas
  • GS1 spray gas
  • GS2 dry gas
  • IS source injection voltage
  • TEM was 400 °C.
  • the scanning ion reaction (MRM) of diclofenac and internal standard was 293.6 ⁇ 249.9, 284.9 ⁇ 185.9, and the collision energy (CE) was -14.8 and -23.8V, respectively.
  • the pharmacokinetic parameters of diclofenac sodium in plasma are shown in Table 18, and the drug-time curve is shown in Figure 5 after the two different preparations of 37.5 mg of diclofenac sodium were administered intravenously.
  • Example 1 is 1.29 times. It can be seen that the diclofenac sodium preparation prepared according to the invention can rapidly reach a peak of the drug, increase the drug exposure, and improve the analgesic effect of the drug.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Dermatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Inorganic Chemistry (AREA)
  • Medicinal Preparation (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

本发明提供了一种供注射用双氯芬酸钠药物组合物，用于治疗轻、中、重度的疼痛，由包含以下组分或优选由下列组分组成的配方配制的溶液经冷冻干燥工艺制备：双氯芬酸钠、水溶性环糊精衍生物、稳定剂和注射用水，其中所述稳定剂可增加组合物的放置稳定性。

Description

供注射用双氯芬酸钠药物组合物及其制备方法 技术领域
本发明属于医药技术领域，具体涉及一种供注射用双氯芬酸钠药物组合物及该制剂的制备方法。
背景技术
双氯芬酸钠(Diclofenac Sodium)属于苯乙酸类衍生物，为第三代强效非甾体抗炎镇痛药，又名双氯灭痛、服他林、阿米雷尔，为白色结晶性粉末，无臭，易溶于丙酮，溶于甲醇和乙醇，微溶于水，有引湿性，其分子式为C14H10Cl2NNaO2，分子量为318。双氯芬酸钠结构式如下：
Figure PCTCN2017070685-appb-000001
双氯芬酸钠主要作用机制为抑制环氧化酶活性，从而阻断花生四烯酸转化为前列腺素。同时，也能促进花生四烯酸与甘油三酯结合，降低胞内游离的花生四烯酸浓度而间接抑制白三烯的合成，从而达到镇痛和消炎的目的。其临床解热镇痛疗效确切，广泛应用于风湿类风湿、腰椎间盘突出、颈椎痛、牙痛等领域。
目前市售双氯芬酸钠的剂型主要为片剂、胶囊剂等常用口服剂型，擦剂、栓剂、凝胶剂等局部用药以及注射液，尚未见双氯芬酸钠的注射用冻干制剂上市。上述市售双氯芬酸钠制剂均存在着明显不足：口服和局部用药剂型在生物利用度和起效速率等方面还不是很理想，而且给药常会引起较严重的胃肠道反应及首过效应。双氯芬酸钠注射液可克服上述不足，但由于双氯芬酸钠在水中溶解度较低(一般在9mg/mL，25℃)，达不到制剂浓度要求，且双氯芬酸钠的结构中含有易被氧化的基团，将其制备成注射液后药物的稳定性下降。目前，诺华制药公司开发的双氯芬酸钠注射液(Voltarol，75mg/3mL)，其给药方式为肌肉注射或静脉滴注，制剂中以丙二醇和苯甲醇作为增溶剂， 对血管有较强刺激性，给药时病人疼痛感强，长期应用易引起静脉炎症，临用前，必须稀释、用碳酸氢钠缓冲，静脉滴注前必须针对每位病人重新配液，并且需要长时间(120-240min)缓慢滴注以降低静脉炎副作用的发生率。双氯芬酸钠环糊精包合注射液(Dyloject，75mg/2mL)经Shimoda Biotech公司许可，由Javelin制药公司采用环糊精技术开发而成，于2007年12月在英国上市，用于治疗急性疼痛，可以肌肉注射或静脉注射两种方式给药。该剂型用羟丙基-β-环糊精包合双氯芬酸钠，有效地解决了主成分溶解度差的问题，并使用了硫代甘油作为抗氧剂防止药物的氧化降解。但2010年5月，该制剂由于某些批次的制剂瓶内发现不明白色颗粒而被全部召回从而暂时撤出英国市场，这与环糊精水溶液对双氯芬酸钠包合长期放置稳定性不佳，药物泄漏析出结晶有关。2014年12月FDA批准Hospira公司37.5mg/mL规格Dyloject上市，用于治疗轻、中、重度疼痛，给药方式为静脉给药，该制剂处方与75mg/2mL规格相同，使用羟丙基-β-环糊精提高药物溶解度(333mg/mL)，使用硫代甘油为抗氧剂(5mg/mL)，注射时间需大于15s，同样存在长期放置稳定性的问题，在其FDA说明书中剂量和用法项下明确规定在使用前应观察是否有肉眼可见微粒和变色情况，如有微粒或变色情况出现则不能使用，说明制剂存在药物析晶和氧化降解的风险。
现有公知技术多选择将供注射用双氯芬酸钠制备成注射液的制剂形式，并在其中加入一定量的表面活性剂、抗氧剂等添加剂，不能解决制剂的刺激性或析晶和氧化降解问题，同时制剂中羟丙基-β-环糊精等増溶物质用量较大，长期应用毒副作用大。专利CN 94118042公开了一种包含双氯芬酸或其盐和2-羟丙基-β-环糊精的可注射药物组合物或兽用组合物，该组合物制备时需对水溶液进行酸化，未添加pH缓冲剂，组合物pH值下限偏酸性，且未对组合物长期放置稳定性进行研究；该组合物中双氯芬酸钠最高浓度仅为25mg/ml，此外溶液中还含有偏亚硫酸氢钠、氯丁醇等防腐剂。中国专利CN 1711996A公开了一种含有双氯芬酸钠和β-环糊精的可注射药物组合物，该组合物为水溶液形式，含有双氯芬酸钠、β-环糊精和聚山梨醇酯，仅适合于皮下和肌肉给药。中国专利CN 200410024064公开了一种含有双氯芬酸盐和利多卡因的冻干制剂及其制备方法，该发明通过双氯芬酸盐和利多卡因两种具有药理活性的成分发挥作用，该技术采用表面活性剂吐温80作为增溶剂，在处方中加入防腐剂，对病人的刺激性和毒副作用较大，未考虑药物的氧化降解问题。 专利CN 101244278A公开了一种可供静脉注射的双氯芬酸钠注射用制剂及其制备方法，该技术采用羟丙基-β-环糊精对双氯芬酸钠进行包合，在制剂中加入抗氧剂，未考虑制剂稳定性和毒副作用问题，该制剂与市售Dyloject相比无明显改进。专利CN 101123957A公开了一种双氯芬酸及其药学上可接受盐的注射剂，该制剂为溶液形式，含醇类共溶剂、抗氧剂、防腐剂等大量添加剂，具有较大副作用，且只能用于肌注和缓慢静脉推注。
发明内容
[技术问题]
为了克服现有供注射用双氯芬酸钠制剂的长期稳定性差、易发生析晶和降解、毒副作用大的问题，本发明采用特定的双氯芬酸钠药物组合物和制备方法，从而获得了稳定安全、质量可靠、毒副作用小的供注射用双氯芬酸钠的药物组合物。
本发明提供的供注射用双氯芬酸钠药物组合物复溶后可直接静脉注射，可用于治疗轻、中、重度的疼痛，其中环糊精衍生物用量低，少量溶剂复溶后与人体血液等渗，可明显减轻注射位点的疼痛，加快药物注射时间(5～10s)，提高镇痛效果；此外，处方中无需添加抗氧剂，还可避免注射液在放置过程中析出结晶的风险，显著增加了制剂稳定性，提高了临床应用的安全性，增加了病人给药的顺应性，创造了更高的临床治疗效益。
[技术方案]
因此，本发明的一个目的在于提供一种稳定性好、有效安全的供注射用双氯芬酸钠药物组合物，该药物组合物含有双氯芬酸钠、水溶性环糊精衍生物、稳定剂。本发明的供注射用双氯芬酸钠药物组合物，是一种冻干粉针剂，由包含双氯芬酸钠、水溶性环糊精衍生物、稳定剂、注射用水的配方配制的溶液经冷冻干燥工艺制成，其中所述溶液含有下列组分或优选由下列组分组成：
Figure PCTCN2017070685-appb-000002
其中，由所述配方配置的溶液经过滤、冷冻干燥制备冻干粉针剂时，将 残氧量控制为低于2％，溶氧量控制为低于1mg/L。
优选地，所述配方含有下列组分或优选由下列组分组成：
Figure PCTCN2017070685-appb-000003
所述水溶性环糊精衍生物可以是选自羟丙基-β-环糊精、磺丁基-β-环糊精和葡糖基-β-环糊精中的至少一种的可注射用的水溶性环糊精衍生物，优选为羟丙基-β-环糊精。
本发明是通过水溶性环糊精衍生物对双氯芬酸钠的包合形成包合物以提高双氯芬酸钠的溶解度，其中，本发明的发明人发现，将双氯芬酸钠与水溶性环糊精衍生物加水溶解后，在双氯芬酸钠浓度为37.5mg/mL的溶液中，维持体系pH范围为7-10，可显著提高双氯芬酸钠的包合程度，这可能是因为碱性环境可影响包合物的立体结构，增加环糊精提供的空洞数，从而促进对双氯芬酸钠的包合。其中，溶液的碱性越强，双氯芬酸钠的包合作用越强，综合考虑最终制剂的刺激性，故而选择维持溶液pH值至7.0-10.0范围内，优选pH值为8.0-9.0。
所述稳定剂为药学上可接受的可注射用盐类，例如，各类缓冲盐体系(如氨丁三醇、磷酸盐(如磷酸氢二钠)、柠檬酸-柠檬酸钠等)，或所述缓冲盐体系与氢氧化钠、盐酸等的组合。优选地，所述稳定剂为氨丁三醇或磷酸盐缓冲体系，或它们与氢氧化钠的组合。
本发明的包含双氯芬酸钠的处方中，水溶性环糊精衍生物与双氯芬酸钠的包合作用呈动态平衡，该平衡受多种因素影响。其中，本发明的发明人发现采用具有缓冲能力的所述稳定剂，可减少或避免制得的冻干制剂再溶解后的析晶现象，并显著提高包合物稳定性。包合物析晶现象的发生可能与冻干制剂再溶解后的溶液放置过程中pH值下降，从而破坏包合物动态平衡有关，动态平衡被破坏后水溶性环糊精衍生物无法完全包合双氯芬酸钠，使得包和物稳定性差，进而导致药物析晶。采用根据本发明的稳定剂控制处方的pH范围，可避免体系pH值发生显著变化，维持包合物的动态平衡状态，减少或避免上述析晶现象，提高包合物稳定性，同时缓冲盐还可与冻干制剂中残留的水分子结合，避免水分子与双氯芬酸钠竞争性结合环糊精衍生物范德华 (Vander Waals)力作用位点。
本发明的另一个目的在于提供一种便于工业化大批量生产、工艺简单的供注射用双氯芬酸钠药物组合物的制备方法，其包括以下步骤：
1)根据上述配方制备溶液：将水溶性环糊精衍生物溶于部分的注射用水中，再加入双氯芬酸钠，搅拌至完全溶解，加入稳定剂搅拌溶解，补加剩余量的注射用水；
2)加入活性炭，继续搅拌15-30min，过滤，制得中间溶液；
3)测定所述中间溶液中双氯芬酸钠的含量；
4)根据步骤3)所述双氯芬酸钠含量的测定结果，向瓶中充入惰性气体，将残氧量控制为低于2％，溶氧量控制为低于1mg/L，按制剂单位灌装(分装)入各个所述瓶中，半加塞；
5)将步骤4)得到的药液冷冻干燥、压塞，在无菌条件下压盖，密封，即得双氯芬酸钠药物组合物。
所述步骤1)中，搅拌时间优选为1-3h，从而完成水溶性环糊精衍生物对双氯芬酸钠的包合，提高双氯芬酸钠的溶解度。
研究发现，溶液的碱性越强，水溶性环糊精衍生物对药物的包合増溶能力越强，综合考虑给药刺激性等因素，所述步骤1)中，优选地，加入所述稳定剂以使得在将所述供注射用双氯芬酸钠药物组合物加水复溶为双氯芬酸钠浓度为37.5mg/mL的溶液时，所述稳定剂可将溶液的pH值范围控制为7-10，优选8.0-9.0。
所述步骤1)中，所述稳定剂为药学上可接受的可注射用盐类，例如，各类缓冲盐体系(如氨丁三醇、磷酸盐(如磷酸氢二钠)、柠檬酸-柠檬酸钠等)，或所述缓冲盐体系与氢氧化钠、盐酸等的组合。优选地，所稳定剂为氨丁三醇或磷酸盐缓冲体系，或它们与氢氧化钠的组合。在这里，具有缓冲能力的偏碱性稳定剂有利于确保药物的稳定包合和保证产品稳定性。
所述步骤2)中，活性炭用量优选为步骤1)所述处方溶液总重的0.02％-0.1％，活性炭用量太少，将不能完全将溶液脱色、去热原和除杂质，用量过多，则会吸附溶液中的活性成分，所以活性炭的用量应控制在合理范围内。
所述步骤4)中，充入惰性气体可用于控制残氧量和溶氧量，保护双氯芬酸不发生氧化降解，提高产品稳定性，所述惰性气体优选为氮气。
所述步骤5)中，所述冷冻干燥工艺为本领域的常规冻干工艺，例如，所述冻干工艺可优选为：将已分装好的瓶放至冻干箱中，降温至-40℃，于-40℃恒温预冻1-3h，然后抽真空；升温至-29℃，保持3-5h；升温至-10℃，保持4-6h；升温至0℃，保持1h；升温至25℃，保持5-8h。但本发明并不仅限于此。
所述步骤5)中，压塞前可选择真空压塞或充入一定压力的惰性气体后压塞，上述两种条件的压塞使得瓶中均无氧存在，这可避免制得的产品在放置过程中发生氧化降解反应。在这里，不能选择充入无菌空气后压塞，氧气的引入会影响本品稳定性。通过长期和加速6月稳定性研究表明产品稳定性良好，氧化降解杂质未增加，组合物外观未发生由氧化反应引起的颜色变化。
所述供注射用双氯芬酸钠药物组合物中活性药物双氯芬酸钠的含量为75mg每瓶或37.5mg每瓶。
所述供注射用双氯芬酸钠药物组合物加水复溶制成含活性药物双氯芬酸钠37.5mg/mL的溶液时，pH范围为7.0-10.0，优选8.0-9.0。
本发明的另一个目的在于提供一种稳定性好、有效安全的供注射用双氯芬酸钠药物组合物，所述双氯芬酸钠药物组合物是一种冻干粉针剂，其由以上方法制备。
本发明的另一个目的在于提供一种用于制备所述的供注射用双氯芬酸钠药物组合物的溶液，其中，所述溶液包含如下组分：双氯芬酸钠；水溶性环糊精衍生物；稳定剂；和注射用水。
本发明的另一目的在于提供所述供注射用的双氯芬酸钠药物组合物用于治疗轻、中、重度的疼痛的用途。
[有益效果]
本发明的供注射用双氯芬酸钠药物组合物及其制备方法，具有如下优点：
1、安全性高，环糊精用量较现有市售品Dyloject显著降低，从而降低肾毒性和溶血等毒副作用；处方溶液中无需使用抗氧剂，降低了安全隐患。制剂中辅料的种类和用量较少，降低了对人体的损害，其安全性符合国家相关法规的要求；
2、所述药物组合物加水复溶后所得的37.5mg/mL双氯芬酸钠浓度的注射液与人体血液等渗，与生理盐水的渗透压摩尔浓度比为0.9～1.1，可直接静脉注射，并可减少注射液对血管的刺激性和病人的疼痛感，而现有市售品 Dyloject(75mg/2mL)与人体不等渗；
3、稳定性好，冻干粉针的制剂形式可避免注射液在长期放置过程中析出晶体沉淀的风险；采用特殊的缓冲体系控制制剂pH值，增强水溶性环糊精衍生物对双氯芬酸钠的包合作用；控制残氧量，氧化反应发生可能性低；冻干制剂中的杂质低于0.2％，显著增加制剂稳定性，使其质量可靠；
4、疗效及病人顺应性良好，复溶后的注射液具有与Dyloject相同的医药性质，且相比于Dyloject需大于15s的注射时间，可进一步加快药物的注射时间(5～10s)，显著加快药物起效时间，可用于治疗轻、中、重度疼痛，快速减轻病人的疼痛，此外还减轻注射位点的疼痛不适感，增加病人给药的顺应性；
5、运输携带和临床应用方便，制备成冻干制剂后，运输更加便捷和安全，减少了损失，降低了运输成本，携带方便，使用便捷；
6、成本低廉，本发明制得的药物组合物中辅料种类少、用量低，制得的冻干制剂却能达到甚至超过其它同类型的药物，充分证明了本发明所述药物的突出优势；
7、本发明的供注射用双氯芬酸钠药物组合物的制备方法工艺简单、高效，成本低廉，适宜工业化应用。
附图说明
图1是显示了根据本发明的制备实施例1的处方(除注射用水外)简单混合的物理混合物的X-射线衍射图。
图2是显示了根据本发明的制备实施例1制备的供注射用双氯芬酸钠药物组合物0个月的X-射线衍射图。
图3是显示了根据本发明的制备实施例1制备的供注射用双氯芬酸钠药物组合物加速6个月的X-射线衍射图。
图4是显示了根据本发明的制备实施例1制备的供注射用双氯芬酸钠药物组合物长期24个月的X-射线衍射图。
图5是采用根据本发明的制备实施例4制备的供注射用双氯芬酸钠药物组合物和对比实施例1制备的双氯芬酸钠药物注射液对犬静脉注射给药后的药时曲线图。
图6是图5所示的药时曲线图中前2h的局部放大图。
具体实施方式
下面结合具体的实施例，进一步阐述本发明。应当理解的是，这些实施例仅用于说明本发明而非以任何方式限制本发明的范围。在以下实施例中，未详细描述的各种过程和方法是本领域中公知的常规方法。
以下实施例中：双氯芬酸钠(安阳九州药业有限责任公司)；羟丙基-β-环糊精(罗盖特(中国)精细化工有限公司)；磺丁基-β-环糊精(美国Captisol公司)；氢氧化钠(湖南尔康制药股份有限公司)；氨丁三醇(武汉顶辉化工有限公司)；磷酸氢二钠(湖南九典制药有限公司)；注射用水(自制)；以及常规未标注试剂购自国药集团化学试剂有限公司。
制备实施例
制备实施例1
Figure PCTCN2017070685-appb-000004
制备方法为：
1)将羟丙基-β-环糊精溶于部分的注射用水中，再加入双氯芬酸钠，搅拌至完全溶解，加入氨丁三醇和氢氧化钠搅拌溶解，补加剩余量的注射用水；
2)加入0.1％的活性炭，搅拌15min，以钛棒过滤器脱炭，再以0.22μm滤膜过滤除菌，制得中间溶液；
3)以HPLC法检测所述中间溶液中双氯芬酸钠含量；
4)向玻璃瓶中充入氮气，控制残氧量1.7％，溶氧量0.7mg/L，根据所述双氯芬酸钠含量的测定结果，将所得药液按制剂单位分装入各个玻璃瓶中，半加塞；
5)将已灌装好的玻璃瓶放至冻干箱中，降温至-40℃，于-40℃恒温预冻2h，然后抽真空；升温至-29℃，保持3h；升温至-10℃，保持4h；升温至0℃，保持1h；升温至25℃，保持5h。充氮气至-0.08MPa后压塞。在无菌条件下压盖，铝封，即得供注射用双氯芬酸钠药物组合物。
制备实施例2
Figure PCTCN2017070685-appb-000005
制备方法为：
1)将磺丁基-β-环糊精溶于部分的注射用水中，再加入双氯芬酸钠，搅拌至完全溶解，加入氨丁三醇搅拌溶解，补加剩余量的注射用水；
2)加入0.05％的活性炭，搅拌20min，以钛棒过滤器脱炭，再以0.22μm滤膜过滤除菌，制得中间溶液；
3)以HPLC法检测所述中间溶液中双氯芬酸钠含量；
4)向玻璃瓶中充入氮气，控制残氧量1.6％，溶氧量0.6mg/L，根据所述双氯芬酸钠含量的测定结果，将所得药液按制剂单位分装入各个玻璃瓶中，半加塞；
5)将已灌装好的玻璃瓶放至冻干箱中，降温至-40℃，于-40℃恒温预冻3h然后抽真空；升温至-29℃，保持5h；升温至-10℃，保持6h；升温至0℃，保持1h；升温至25℃，保持8h。充氮气至-0.06MPa后压塞。在无菌条件下压盖，铝封，即得供注射用双氯芬酸钠药物组合物。
制备实施例3
Figure PCTCN2017070685-appb-000006
制备方法为：
1)将葡糖基-β-环糊精溶于部分的注射用水中，再加入双氯芬酸钠，搅拌至完全溶解，加入磷酸氢二钠搅拌溶解，补加剩余量的注射用水；
2)加入0.05％的活性炭，搅拌30min，以钛棒过滤器脱炭，再以0.22μm滤膜过滤除菌，制得中间溶液；
3)以HPLC法检测所述中间溶液中双氯芬酸钠含量；
4)向玻璃瓶中充入氮气，控制残氧量1.5％，溶氧量0.5mg/L，根据所 述双氯芬酸钠含量的测定结果，将所得药液按制剂单位分装入各个玻璃瓶中，半加塞；
5)将已灌装好的玻璃瓶放至冻干箱中，降温至-40℃，于-40℃恒温预冻2h，然后抽真空；升温至-29℃，保持4h；升温至-10℃，保持5h；升温至0℃，保持1h；升温至25℃，保持5h。在真空条件下压塞。在无菌条件下压盖，铝封，即得供注射用双氯芬酸钠药物组合物。
制备实施例4
Figure PCTCN2017070685-appb-000007
制备方法为：
1)将羟丙基-β-环糊精溶于部分的注射用水中，再加入双氯芬酸钠，搅拌至完全溶解，加入磷酸氢二钠和氢氧化钠搅拌溶解，补加剩余量的注射用水；
2)加入0.1％的活性炭，搅拌15min，以钛棒过滤器脱炭，再以0.22μm滤膜过滤除菌，制得中间溶液；
3)以HPLC法检测所述中间溶液中双氯芬酸钠含量；
4)向玻璃瓶中充入氮气，控制残氧量1.6％，溶氧量0.5mg/L，根据所述双氯芬酸钠含量的测定结果，将所得药液按制剂单位分装入各个玻璃瓶中，半加塞；
5)将已灌装好的玻璃瓶放至冻干箱中，降温至-40℃，于-40℃恒温预冻2h，然后抽真空；升温至-29℃，保持3h；升温至-10℃，保持4h；升温至0℃，保持1h；升温至25℃，保持5h。充氮气至-0.08MPa后压塞。在无菌条件下压盖，铝封，即得供注射用双氯芬酸钠药物组合物。
对比实施例
对比实施例1(Dyloject制剂)
Figure PCTCN2017070685-appb-000008
Figure PCTCN2017070685-appb-000009
制备方法为：将羟丙基-β-环糊精溶于部分(约90％)的注射用水中，再加入双氯芬酸钠，搅拌至完全溶解，加入氢氧化钠搅拌溶解，补加剩余量(约10％)的注射用水；再以0.22μm滤膜过滤除菌，按1mL/瓶灌装、加塞、压盖，于116℃灭菌15min，即得双氯芬酸钠注射液。
本发明人发现，采用市售Dyloject处方制备的双氯芬酸钠注射液，在高温灭菌步骤时制剂产生大量降解物质(由0.05％增至1.27％)，表明本品对高温敏感。本发明的制备方法无需注射液的高温灭菌步骤，可显著减少本品的杂质量，提高稳定性。
对比实施例2
Figure PCTCN2017070685-appb-000010
制备方法为：
1)将羟丙基-β-环糊精溶于部分(约90％)的注射用水中，再加入双氯芬酸钠，搅拌至完全溶解，加入氢氧化钠搅拌溶解，补加剩余量(约10％)的注射用水；
2)加入0.05％的活性炭，搅拌15min，以钛棒过滤器脱炭，再以0.22μm滤膜过滤除菌，制得中间溶液；
3)以HPLC法检测所述中间溶液中双氯芬酸钠含量；
4)向玻璃瓶中充入氮气，控制残氧量1.7％，溶氧量0.7mg/L，根据所述双氯芬酸钠含量的测定结果，将所得药液按制剂单位分装入各个玻璃瓶中，半加塞；
5)将已灌装好的玻璃瓶放至冻干箱中，降温至-40℃，于-40℃恒温预冻1h，然后抽真空；升温至-29℃，保持3h；升温至-10℃，保持4h；升温至0℃，保持1h；升温至25℃，保持5h。充氮气至-0.08MPa后压塞。在无菌 条件下压盖，铝封，即得。
对比实施例3
Figure PCTCN2017070685-appb-000011
制备方法为：
1)将羟丙基-β-环糊精溶于部分的注射用水中，再加入双氯芬酸钠，搅拌至完全溶解，加入磷酸氢二钠搅拌溶解，补加剩余量的注射用水；
2)加入0.05％的活性炭，搅拌15min，以钛棒过滤器脱炭，再以0.22μm滤膜过滤除菌，制得中间溶液；
3)以HPLC法检测所述中间溶液中双氯芬酸钠含量；
4)根据所述双氯芬酸钠含量的测定结果，将所得药液按制剂单位分装入各个玻璃瓶中，半加塞；
5)将已灌装好的玻璃瓶放至冻干箱中，降温至-40℃，于-40℃恒温预冻1h，然后抽真空；升温至-29℃，保持3h；升温至-10℃，保持4h；升温至0℃，保持1h；升温至25℃，保持5h。充无菌空气至-0.08MPa后压塞。在无菌条件下压盖，铝封，即得。
实验实施例
实验实施例1(复溶稳定性)
取制备实施例1所得的供注射用双氯芬酸钠药物组合物若干瓶，每瓶加1ml水复溶，考察制剂的复溶稳定性。考察时间为0h、2h、4h、8h、12h、24h，考察指标为澄清度、碱度、含量、有关物质、可见异物和不溶性微粒，测定结果见表1。
表1供注射用双氯芬酸钠药物组合物复溶稳定性考察结果
Figure PCTCN2017070685-appb-000012
试验结果表明：本发明的供注射用双氯芬酸钠药物组合物复溶后24h内稳定性良好。
实验实施例2(制剂稳定性)
按照国家食品药品监督管理局发布的化学药物(原料药和制剂)稳定性研究技术指导原则进行稳定性试验，取制备实施例1、制备实施例2制备的供注射用双氯芬酸钠药物组合物和对比实施例1制得的双氯芬酸钠注射液(与市售Dyloject处方相同)、对比实施例2、对比实施例3制得的双氯芬酸钠冻干制剂，进行加速(40℃±2℃/75％RH±5％RH)6个月和长期(25℃±2℃/60％RH±5％RH)24个月试验研究，对样品的关键指标：性状、澄清度、碱度(加注射用水复溶制成含双氯芬酸钠37.5mg/ml的溶液)、可见异物、不溶性微粒、有关物质及含量进行检验，检测结果见表2。
表2供注射用双氯芬酸钠药物组合物稳定性考察结果
Figure PCTCN2017070685-appb-000013
由结果可见，制备实施例1和2制备的产品在加速、长期试验中各项关键指标无明显变化，样品外观未发生任何颜色变化，复溶后可见异物和不溶性微粒均符合规定，复溶后样品24h内稳定性良好，未出现任何析晶现象，杂质未出现显著增加；而对比实施例1制备的双氯芬酸钠注射液在放置过程中出现pH下降、有关物质显著增加的现象，加速条件样品出现析出物、颜色发生显著变化；对比实施例2的未加稳定剂的双氯芬酸钠冻干制剂在放置过程中出现pH下降、有关物质显著增加的现象，加速条件下样品复溶后可见异物，从而不符合规定；以及对比实施例3的未进行溶氧、残氧控制且未在惰性气体或真空条件下压塞的双氯芬酸钠冻干制剂在放置过程中出现pH下降、有关物质显著增加的现象，加速条件下样品复溶后可见异物，从而不符合规定。可见本发明制得的供注射用双氯芬酸钠药物组合物具有质量保证，且稳定性良好。
实验实施例3
取制备实施例1所制得的供注射用双氯芬酸钠药物组合物0个月、加速6个月和长期24个月的稳定性样品以及按制备实施例1的处方(除注射用水外)简单混合的物理混合物粉末，进行X-射线衍射检测，考察羟丙基-β-环糊精对药物的包合情况。检测条件为3.0～40.0°，0.01°/秒，加速电压：40KV，电流40mA。检测结果见图1～图4。结果表明，制备实施例1中双氯芬酸钠均以无定型形式存在，羟丙基-β-环糊精对双氯芬酸钠包合完全，药物放置过程中稳定性良好。
实验实施例4
取本发明的制备方法获得的供注射用双氯芬酸钠药物组合物进行特殊安全性试验：溶血性试验、刺激性试验和过敏性试验。具体操作为：取本发明制备实施例4制得的供注射用双氯芬酸钠药物组合物每瓶加1ml注射用水复溶，制得浓度为37.5mg/mL的受试液，进行动物溶血性、刺激性和过敏性试验。以对比实施例1制得的双氯芬酸钠注射液为对照制剂。
1、溶血性试验
2％兔红细胞混悬液制备：取试验兔(动物：家兔，来源：上海药物研究所实验动物中心)一只，清醒状态下心室内取血约20ml，用棉签搅拌约10分钟以除去纤维蛋白原，再倒入试管中加约10倍量的0.9％氯化钠注射液摇 匀，1,500转/分钟离心15分钟，除去上清液，沉淀的红细胞再用0.9％氯化钠注射液洗涤离心3次以上，至上清液不呈红色为止。将所得红细胞用0.9％氯化钠注射液配成2％红细胞混悬液，供试验用。
取二批清洁试管各7支，按表3配比量依次加入2％红细胞混悬液、0.9％氯化钠注射液或蒸馏水和37.5mg/ml受试液，混匀后置于37℃±0.5℃的恒温水浴箱中进行温育，开始时每隔15分钟观察1次，1小时后每隔1小时观察1次，观察3个小时内各试管溶液有无溶血及红细胞凝聚作用，判断标准见表4。
表3供注射用双氯芬酸钠药物组合物的溶血性试验
Figure PCTCN2017070685-appb-000014
表4溶血判断标准
Figure PCTCN2017070685-appb-000015
如出现红细胞凝聚现象，可按下法进一步判定是真凝聚还是假凝聚。若凝聚物在振荡后以能均匀分散，或将凝聚物放在载玻片上，在盖玻片边缘加2滴0.9％氯化钠溶液，置显微镜下观察，凝聚红细胞能被冲散者为假凝聚，相反，凝聚物不被摇散或在玻片上不被冲散者为真凝聚。
结果判断：当6号试管(阴性对照管)无溶血和凝聚现象，7号试管(阳性对照管)有溶血现象时，若1-5号试管中溶液在3小时内不发生溶血和凝聚现象，则试验药物可供注射使用；若1-5号试管中溶液在3小时内发生溶血和凝聚现象，则试验药物不可供注射使用。
溶血试验结果为：7号试管加入蒸馏水后即刻呈红色澄明，管底无红细胞残留，表明全部溶血。1-6号试管溶液15min时红细胞开始下沉，随时间延 长则下沉红细胞逐渐增多，上清液逐渐增多，上清液呈淡黄色澄明，表明受试液和对照品氯化钠注射液均无溶血作用，3小时后振摇各试管，1-6号试管红细胞均可均匀散开，无红细胞凝聚现象。具体试验结果见表5。
表5供注射用双氯芬酸钠药物组合物的溶血试验结果
Figure PCTCN2017070685-appb-000016
结果表明本发明的供注射用氯芬酸钠药物组合物在试管内的最高终浓度为37.5mg/ml时对兔红细胞无溶血或凝聚现象。
2、刺激性试验
剂量设计：双氯芬酸钠临床推荐剂量为37.5mg/次，折算为兔用剂量为1.9mg/kg，因此兔试验剂量确定为2mg/kg。
取试验兔(动物：家兔，来源：上海药物研究所实验动物中心)8只，分为实验组和对照组2组，4只/组，雌雄各半，分别用兔固定箱固定，左右耳缘去毛，酒精消毒，从耳缘静脉最远心端开始进针，试验组左右两侧耳缘静脉分别静脉注射0.9％氯化钠注射液(对照侧)和本发明的供注射用双氯芬酸钠药物组合物的受试液(给药侧)，静脉注射量为2mg/kg体重，注射速度约为5～10s内完成注射；对照组左右两侧耳缘静脉分别静脉注射0.9％氯化钠注射液(对照侧)和对比实施例1制得的双氯芬酸钠对照制剂(给药侧)，静脉注射量为2mg/kg体重，注射速度约为大于15s完成注射。各组每日进针一次，连续5天，每日给药部位由耳缘静脉远心端向近心端逐次前移，肉眼观察兔耳缘静脉反应，包括血管是否清晰，有无血管扩张或收缩，血管内是否有淤血，血管周围组织有无水肿等。
每组试验兔于末次给药后48小时将二只试验兔放血处死，取下两耳，在 末次注射部位的近心端附近(距末次注射部位约1cm，a段)和近心端远侧(距末次注射部位约3cm，b段)各取一段标本，首先进行肉眼观察，判断药物对血管壁有无刺激性，观察标准见表6，判断标准见表7。然后用4％甲醛固定，进行组织病理学检查，检查标准见表8，判断标准见表9。末次给药后96小时将每组一只试验兔放血处死，按上述方法进行检查。末次给药后14天将每组最后一只试验兔放血处死，按上述方法进行检查。
表6血管刺激性试验肉眼目检观察评分标准
Figure PCTCN2017070685-appb-000017
表7血管刺激性试验肉眼目检刺激强度判断标准
Figure PCTCN2017070685-appb-000018
表8血管刺激性试验显微镜检查评分标准
Figure PCTCN2017070685-appb-000019
表9血管刺激性试验显微镜检查刺激强度判断标准
Figure PCTCN2017070685-appb-000020
给药期间，每天给药后1小时观察，实验组的试验兔给药侧耳缘静脉血管纹路清晰，无血管扩张或收缩，血管内无明显淤血，血管周围组织无水肿，较对照侧耳缘静脉有明显改善。
选取兔耳缘静脉标本前观察，实验组的试验兔给药侧耳缘静脉均未见明显充血、水肿症状，评分值均为0，较对照侧耳缘静脉有明显改进。结果见表10。
表10血管刺激性试验肉眼目检观察结果
Figure PCTCN2017070685-appb-000021
组织切片检查结果表明，实验组末次给药后48小时、96小时及14天，试验兔给药侧耳缘静脉a、b段血管结构均基本正常，无明显血管扩张充血现象，无明显血栓形成，血管内皮细胞基本正常，无肿胀、坏死脱落等现象，血管周围组织无水肿，无炎细胞浸润，平均分值均为0，与对照侧比较无明显差异。对照组末次给药后48小时、96小时及14天，试验兔给药侧耳缘静脉a、b段血管结构均基本正常，部分出现血管扩张充血现象，无明显血栓形成，血管内皮细胞基本正常，无肿胀、坏死脱落等现象，血管周围组织部分出现水肿，无炎细胞浸润，平均分值均为0.75，表现出轻度刺激性。
上述试验结果表明，制备实施例4制备的双氯芬酸钠受试液37.5mg/ml耳缘静脉注射，每日一次，连续5天，对兔耳缘静脉未见明显局部刺激反应。 按市售处方制备的对比实施例1的对照制剂37.5mg/ml耳缘静脉注射，每日一次，连续5天，对兔耳缘静脉表现出轻度刺激反应。可见，本发明的双氯芬酸钠药物组合物对刺激性有显著改善，安全性良好。
3、主动过敏性试验
剂量设计：双氯芬酸钠临床推荐剂量为37.5mg/次，折算为豚鼠用剂量为2.9mg/kg，因此豚鼠全身主动过敏试验剂量低剂量确定为3.0mg/kg，高剂量确定为6.0mg/kg。
豚鼠(动物：豚鼠，来源：上海药物研究所实验动物中心)24只，按体重性别随机分为4组，每组6只，雌雄各半。第一组作为阴性对照组，腹腔注射0.9％氯化钠注射液0.2ml/100g，隔日注射一次，共3次。第二组作为阳性对照组，腹腔注射5％新鲜鸡蛋清溶液0.2ml/100g，隔日注射一次，共3次。第三组作为低剂量给药组，腹腔注射双氯芬酸钠试验制剂低剂量溶液(取37.5mg/ml双氯芬酸钠受试液1ml置于25ml容量瓶中，加0.9％氯化钠注射液溶解定容)0.2ml/100g，隔日注射一次，共3次。第四组作为高剂量给药组，腹腔注射双氯芬酸钠试验制剂高剂量溶液(取37.5mg/ml双氯芬酸钠受试液2ml置于25ml容量瓶中，加0.9％氯化钠注射液溶解定容)0.2ml/100g，隔日注射一次，共3次。各组豚鼠在末次致敏后第14天及第21天各取3只豚鼠分别自足跖静脉或耳静脉注射2倍量的原药液进行激发，观察3小时内豚鼠是否有抓鼻、喷嚏、竖毛、抽搐等过敏症状，按表11、表12标准判断过敏反应发生程度，计算过敏反应发生率。
激发注射后，若发现过敏反应症状时，可取健康未致敏豚鼠2只，自静脉注射激发剂量药物，观察有无由于受试药物作用引起的类似过敏反应症状。
表11豚鼠过敏反应症状
0正常 7呼吸急促 14步态不稳
1躁动 8排尿 15跳跃
2竖毛 9排粪 16喘息
3颤抖 10流泪 17痉挛
4搔鼻 11呼吸困难 18旋转
5喷嚏 12哮鸣音 19潮式呼吸
6咳嗽 13紫癜 20死亡
表12主动全身过敏性评价标准
0 - 过敏反应阴性
1-4症状 + 过敏反应弱阳性
5-10症状 ++ 过敏反应阳性
11-19症状 +++ 过敏反应强阳性
20 ++++ 过敏反应极强阳性
首次致敏、末次致敏及激发当日各组豚鼠体重比较均无明显差异，结果见表13。激发给药后3小时内，阴性对照组、低剂量给药组、高剂量给药组均未见明显过敏症状，反应均为阴性；阳性对照组豚鼠于激发注射后迅速出现躁动、竖毛、颤抖、搔鼻、喷嚏、咳嗽、呼吸急促、步态不稳、喘息、痉挛、排尿、排粪、潮式呼吸等明显过敏反应甚至死亡，死亡时间均在激发注射后3分钟内，致敏率为100％，反应为极强阳性(结果见表14)。
表13豚鼠平均体重(n＝6)
Figure PCTCN2017070685-appb-000022
表14主动全身过敏反应试验结果
Figure PCTCN2017070685-appb-000023
上述试验结果表明，在本试验条件下，本发明的供注射用双氯芬酸钠药物组合物对豚鼠无明显主动全身致敏作用。
4、被动过敏试验
抗血清制备：取豚鼠8只，按体重性别随机分为4组，每组2只，雌雄各半。第一组作为阴性对照组，腹腔注射0.9％氯化钠注射液0.2ml/100g，隔 日注射一次，共5次。第二组作为阳性对照组，腹腔注射2mg/kg卵蛋白溶液0.2ml/100g，隔日注射一次，共5次。第三组作为低剂量给药组，腹腔注射上述双氯芬酸钠试验制剂低剂量溶液0.2ml/100g，隔日注射一次，共5次。第四组作为高剂量给药组，腹腔注射上述双氯芬酸钠试验制剂高剂量溶液0.2ml/100g，隔日注射一次，共5次。各组豚鼠在末次致敏后第14天股动脉取血，室温自然凝固，2000rpm离心20min分离血清后按分组一同贮存于-18℃冰箱内备用。
被动皮肤过敏试验：取豚鼠24只，按体重性别随机分为4组，每组6只，雌雄各半，分别为阴性对照组、阳性对照组、低剂量给药组、高剂量给药组。用动物脱毛器将豚鼠背部皮肤脱毛，同时用0.9％氯化钠注射液分别配制上述各组抗血清稀释液(1:2、1:8、1:32，v/v)，各组豚鼠自脱毛处背部分别皮内注射相应的抗血清稀释液，每点注射0.1ml，二点之间间隔约2.5cm。激发注射：24小时后按致敏时同样方法配制各组相应溶液，并与1％伊文思兰溶液按1:1比例混合后，静脉注射给药0.4ml/100g。激发注射30min后放血处死豚鼠，剪取背部皮肤，用游标卡尺测定皮肤内层的斑点直径，不规则斑点的直径为长径与短径之和的一半，直径大于5mm者为阳性，并计算阳性反应百分率。
激发给药30min后，阴性对照组、低剂量给药组、高剂量给药组的豚鼠背部皮肤内层的蓝斑直径均小于5mm，反应均为阴性；阳性对照组豚鼠激发给药30min后，背部皮肤内层的蓝斑直径均大于5mm，反应均为阳性，致敏率为100％，反应为极强阳性，结果见表15。
表15被动皮肤过敏反应试验结果(n＝6)
Figure PCTCN2017070685-appb-000024
上述试验结果表明，在本试验条件下，本发明的供注射用双氯芬酸钠药物组合物对豚鼠无明显被动皮肤致敏作用。
实验实施例5
取制备实施例4所制得的供注射用双氯芬酸钠药物组合物，每瓶加1ml水复溶，制得浓度为37.5mg/ml的供试液，在疼痛动物模型中考察药物的镇痛效果。以对比实施例1所制得的双氯芬酸钠注射液为对照液。分别量取供试液和对照液各1ml分别置于50ml容量瓶中，加生理盐水定容，其浓度均被稀释为0.75mg/ml。取健康的昆明系小鼠(动物：小鼠，来源：上海药物研究所实验动物中心)30只，体重20g左右，雌雄各半，随机分为3组，按7.5mg/kg尾静脉注射小鼠，各组小鼠于给药后10min腹腔注射0.6％冰醋酸溶液0.2mL，立即计时，观察15min内小鼠的扭体反应次数，计算扭体反应的抑制率。具体分组及给药方式见表16，试验结果见表17。
表16试验分组及处理(n＝10)
Figure PCTCN2017070685-appb-000025
表17各组小鼠扭体反应结果(n＝10)
Figure PCTCN2017070685-appb-000026
注：“*”表示与空白对照组比较P＜0.01。
由试验结果可见，制备实施例4和对比实施例1的试验组的小鼠扭体反应与空白对照组相比均显著减少，其中制备实施例4为根据本发明制备的溶液，对比实施例1为根据市售Dyloject配制的溶液，因此，两种双氯芬酸钠制剂均对小鼠疼痛反应有显著缓解作用。
实验实施例6
取制备实施例4所制得的供注射用双氯芬酸钠药物组合物，每瓶加1ml水复溶，制得浓度为37.5mg/ml的供试液，以犬为动物模型考察药物的药代动力学行为。以对比实施例1所制得的双氯芬酸钠注射液为对照液。
比格犬3只(动物：犬，来源：上海药物研究所实验动物中心)，雄性，体重10～12kg。受试动物在试验日前3-7天应在试验场所进行适应性饲养。采用自身对照交叉试验设计，分别用制备实施例4加水复溶后的供试液和对比实施例1的注射液配制给药溶液。给药剂量为37.5mg/只。两周期间间隔不少于5天。制备实施例4的给药溶液在5～10s内完成注射，对比实施例1的给药溶液注射时间大于15s。给药前(0h)及给药后2min、5min、15min、30min、1h、2h、4h、6h、8h和12h的时间点经四肢静脉取静脉血1mL，置于EDTA抗凝试管中，3500rpm离心10min(4℃)，分离血浆，-70℃冰箱中冷冻保存待测。
以LC-MS测定血浆中药物浓度。色谱条件：色谱柱选用Agilent XDB C18(150mm×4.6mm，5μm)，流动相为乙腈-水(含4mM乙酸铵，0.08％甲酸)(75:25，V/V)，流速为0.8mL·min-1，柱温25℃，进样体积为5μL。质谱条件：电喷雾离子源(ESI)，气帘气(CUR)压力为20psi，喷雾气(GS1)压力为55psi，干燥气(GS2)压力为55psi，源喷射电压(IS)为-4000V，离子源温度(TEM)为400℃。双氯芬酸及内标的扫描离子反应(MRM)分别为293.6→249.9、284.9→185.9，碰撞能量(CE)分别为-14.8、-23.8V。
比格犬分别静脉注射给予37.5mg双氯芬酸钠的两种不同制剂后，血浆中双氯芬酸钠的药动学参数见表18，药时曲线见图5。
表18比格犬经静脉注射不同制剂后的药动学参数
Figure PCTCN2017070685-appb-000027
由试验结果可知，与对比实施例1相比，本发明的制备实施例4制备的制剂药物达峰时间由4min减至2min，Cmax值为对比实施例1的1.28倍，AUC值为对比实施例1的1.29倍。可见，根据本发明制备的双氯芬酸钠制剂可使药物快速达峰，增加药物暴露量，提高药物镇痛效果。

Claims (9)

  1. 一种供注射用双氯芬酸钠药物组合物，其特征在于，所述药物组合物为由包含下列组分的配方配置的溶液经冷冻干燥制备的冻干粉针剂，
    Figure PCTCN2017070685-appb-100001
    其中，由所述配方配置的溶液经冷冻干燥制备冻干粉针剂时，将残氧量控制为低于2％，溶氧量控制为低于1mg/L。
  2. 根据权利要求1所述的药物组合物，其中，所述配方包含下列组分：
    Figure PCTCN2017070685-appb-100002
  3. 根据权利要求1所述的药物组合物，其中，所述水溶性环糊精衍生物为选自羟丙基-β-环糊精、磺丁基-β-环糊精和葡糖基-β-环糊精中的至少一种，
    优选地，所述水溶性环糊精衍生物为羟丙基-β-环糊精。
  4. 根据权利要求1所述的药物组合物，其中，所述稳定剂为氨丁三醇、磷酸盐或柠檬酸-柠檬酸钠缓冲体系，或它们与氢氧化钠或盐酸的组合，
    优选地，所述稳定剂为氨丁三醇或磷酸盐缓冲体系，或它们与氢氧化钠的组合。
  5. 根据权利要求1至4中任一项所述的药物组合物，其中，在将所述药物组合物加水复溶为双氯芬酸钠浓度为37.5mg/mL的复溶溶液时，所述稳定剂将所述复溶溶液的pH值范围控制为7-10，优选8.0-9.0。
  6. 一种供注射用双氯芬酸钠药物组合物的制备方法，其包括以下步骤：
    1)根据权利要求1至5中任一项所述的配方制备溶液：将水溶性环糊精衍生物溶于部分的注射用水中，再加入双氯芬酸钠，搅拌至完全溶解，加入稳定剂搅拌溶解，补加剩余量的注射用水；
    2)加入活性炭，继续搅拌15-30min，过滤，制得中间溶液；
    3)测定所述中间溶液中双氯芬酸钠的含量；
    4)根据步骤3)所述双氯芬酸钠含量的测定结果，向瓶中充入惰性气体， 将残氧量控制为低于2％，溶氧量控制为低于1mg/L，按制剂单位分装入各个所述瓶中，半加塞；
    5)将步骤4)得到的药液冷冻干燥、压塞，在无菌条件下压盖，密封，即得所述双氯芬酸钠药物组合物。
  7. 根据权利要求6所述的制备方法，其中，所述步骤1)中，搅拌时间为1-3h。
  8. 根据权利要求6所述的制备方法，其中，所述步骤2)中，活性炭用量为所述处方溶液总重的0.02％-0.1％。
  9. 一种用于制备权利要求1至5中任一项所述的供注射用双氯芬酸钠药物组合物的溶液，其中，所述溶液包含如下组分：
    双氯芬酸钠；
    水溶性环糊精衍生物；
    稳定剂；和
    注射用水。
PCT/CN2017/070685 2016-02-25 2017-01-10 供注射用双氯芬酸钠药物组合物及其制备方法 WO2017143877A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201610104900.XA CN107126418B (zh) 2016-02-25 2016-02-25 供注射用双氯芬酸钠药物组合物及其制备方法
CN201610104900.X 2016-02-25

Publications (1)

Publication Number Publication Date
WO2017143877A1 true WO2017143877A1 (zh) 2017-08-31

Family

ID=59685808

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/070685 WO2017143877A1 (zh) 2016-02-25 2017-01-10 供注射用双氯芬酸钠药物组合物及其制备方法

Country Status (2)

Country Link
CN (1) CN107126418B (zh)
WO (1) WO2017143877A1 (zh)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113456654A (zh) * 2020-07-20 2021-10-01 上海和黄药业有限公司 一种稳定的药物组合物
CN114636769A (zh) * 2022-03-31 2022-06-17 珠海润都制药股份有限公司 一种双氯芬酸钠原料药中杂质的检测方法

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114042048B (zh) * 2021-10-29 2023-02-28 海南海灵化学制药有限公司 一种注射用盐酸头孢吡肟的制备工艺

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1711996A (zh) * 2004-06-22 2005-12-28 Ibsa生物化学研究股份有限公司 含有双氯芬酸钠和β－环糊精的可注射药物组合物
CN101244278A (zh) * 2008-03-13 2008-08-20 北京阜康仁生物制药科技有限公司 一种可供静脉注射的双氯芬酸钠注射用制剂及其制备方法

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0616547A (ja) * 1992-07-01 1994-01-25 Wakamoto Pharmaceut Co Ltd 消炎点眼剤
CN1154070A (zh) * 1994-05-27 1997-07-09 荷兰发马克有限公司 药物组合物
KR20150050595A (ko) * 2006-03-28 2015-05-08 자블린 파머슈티칼스 인코포레이티드 저 복용량의 디클로페낙 및 베타-사이클로덱스트린 제형
CN102871995A (zh) * 2012-10-31 2013-01-16 江苏汉晨药业有限公司 一种含双氯芬酸钠的药物组合物及其制备方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1711996A (zh) * 2004-06-22 2005-12-28 Ibsa生物化学研究股份有限公司 含有双氯芬酸钠和β－环糊精的可注射药物组合物
CN101244278A (zh) * 2008-03-13 2008-08-20 北京阜康仁生物制药科技有限公司 一种可供静脉注射的双氯芬酸钠注射用制剂及其制备方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ZHANG, MEIJUN ET AL.: "The preparation of Inclusion Compound of Diclofenac Sodium- beta -cyclodextrin", STRAIT PHARMACEUTICAL JOURNAL, vol. 20, no. 10, 15 October 2008 (2008-10-15) *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113456654A (zh) * 2020-07-20 2021-10-01 上海和黄药业有限公司 一种稳定的药物组合物
CN113456654B (zh) * 2020-07-20 2023-01-10 上海和黄药业有限公司 一种稳定的药物组合物
CN114636769A (zh) * 2022-03-31 2022-06-17 珠海润都制药股份有限公司 一种双氯芬酸钠原料药中杂质的检测方法

Also Published As

Publication number Publication date
CN107126418B (zh) 2020-05-29
CN107126418A (zh) 2017-09-05

Similar Documents

Publication Publication Date Title
EP4252844A2 (en) Topical compositions and methods for treating inflammatory skin diseases
KR101593579B1 (ko) 치료용 조성물
CA2595617C (en) Formulations for injection of catecholic butanes, including ndga compounds, into animals
KR102465046B1 (ko) 5-콜레스텐-3, 25-디올, 3-술페이트 (25hc3s) 또는 이의 약학적으로 허용 가능한 염, 및 적어도 하나의 시클릭 올리고당을 포함하는 조성물
CN102145003B (zh) 含难溶性药物的药用组合物
BR112015012162B1 (pt) formulação de suspensão tópica e formulação de suspensão de minociclina tópica para usar no tratamento de acne vulgaris
JP2014533251A (ja) メラトニンベースの溶液およびそれらの製造のための粉末
JP5897164B2 (ja) 外用組成物
CN110870914A (zh) 氨基酸类营养素的应用以及包含它的药物组合物
BR112020008077A2 (pt) composição farmacêutica, composição farmacêutica para aplicação tópica e uso de uma composição tópica
WO2017143877A1 (zh) 供注射用双氯芬酸钠药物组合物及其制备方法
JP2021152080A (ja) 医薬組成物
CN110870869A (zh) 包含糖类营养素和常规无效化合物的药物组合物及其应用
CN111825548B (zh) 一种含有芳基丙酸类化合物的药物组合物
JP2019006737A (ja) 医薬組成物
JP2018108955A (ja) 水性製剤
WO2017177160A1 (en) Formulation for use in a method of treatment of pain
US20160128965A1 (en) Gel compositions
CN110870858A (zh) 包含有机酸酸化剂和常规无效化合物的药物组合物及其应用
US20160271092A1 (en) Topical pharmaceutical composition of acitretin
JP7396598B2 (ja) 皮膚創傷用組成物
JP6908449B2 (ja) 医薬組成物
JP2019006697A (ja) 活性酸素消去剤
CN112439066A (zh) 包含化学消融剂和pH调节剂的药物组合物及其应用
CN106937944A (zh) 一种注射用甲硝唑冻干粉针制剂及其制备方法

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17755706

Country of ref document: EP

Kind code of ref document: A1

122 Ep: pct application non-entry in european phase

Ref document number: 17755706

Country of ref document: EP

Kind code of ref document: A1