CN112516093A - Aminomethylbenzoic acid freeze-dried powder injection for injection - Google Patents
Aminomethylbenzoic acid freeze-dried powder injection for injection Download PDFInfo
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Abstract
The invention relates to an aminomethylbenzoic acid freeze-dried powder injection for injection, which comprises aminomethylbenzoic acid, a freeze-dried excipient and an optional acid-base regulator, wherein the freeze-dried excipient is selected from sucrose, glucose, mannitol, lactose, sorbitol and glycine; the pH adjusting agent is selected from sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, hydrochloric acid, phosphoric acid, nitric acid, sulfuric acid, or a combination thereof. The quality detection method comprises the operation of determining the content of impurity p-toluic acid in the freeze-dried powder injection by using high performance liquid chromatography. The method can accurately determine related substances in the freeze-dried powder injection, and the powder injection has excellent pharmaceutical properties such as excellent stability and preparation safety.
Description
Technical Field
The invention belongs to the technical field of medicines, and relates to a freeze-dried powder injection medicinal composition which can be used for treating hemorrhage caused by primary fibrinolysis excessive, including acute and chronic, localized or systemic high-fiber hemorrhage, and the latter is commonly seen in cancer, leukemia, gynecological accidents, severe liver disease hemorrhage and the like, in particular to a freeze-dried powder injection medicinal composition containing aminomethylbenzoic acid, more particularly to a quality detection method of freeze-dried aminomethylbenzoic acid powder injection for injection. The freeze-dried powder injection pharmaceutical composition has expected good properties.
Background
Aminomethylbenzoic Acid (aminotoluic Acid), which is p-Aminomethylbenzoic Acid monohydrate, also known as antifibrinolytic aromatic Acid, p-carboxybenzylamine, 4- (aminomethyl) benzoic Acid, 4-Aminomethylbenzoic Acid, p-Aminomethylbenzoic Acid; 4- (Amino methyl) benzoic acid, 4- (Amino) benzoic acid, CAS number 56-91-7. It is suitable for treating abnormal hemorrhage during operation of lung, liver, pancreas, prostate, thyroid, adrenal gland, gynecological hemorrhage, puerperal hemorrhage, hemoptysis due to pulmonary tuberculosis, bloody sputum, hematuria, prostatic hypertrophy hemorrhage, upper gastrointestinal hemorrhage, etc. The molecular formula of aminomethylbenzoic acid is C8H9NO 2. H2O, the molecular weight is 169.18, and the chemical structural formula is as follows:
aminomethylbenzoic acid is white or off-white flaky crystal or crystalline powder; no smell, slightly bitter taste. Aminomethylbenzoic acid dissolves in boiling water, slightly dissolves in water, and hardly dissolves in ethanol, chloroform, ether or benzene.
Aminomethylbenzoic acid is an aminomethylbenzoic acid type antifibrinolytic agent. Has the same action mechanism as 6-amino acetic acid and tranexamic acid. The three-dimensional configuration of the fibrinolysin is similar to that of lysine, and the fibrinolysin can competitively inhibit the adsorption of the fibrinolysin and protect the fibrin from being degraded by the fibrinolysin, thereby achieving the hemostasis effect. The anti-fibrinolytic activity of the compound is 4 to 5 times stronger than that of 6-aminocaproic acid. Aminomethylbenzoic acid is a procoagulant agent. Various natural antagonists of plasmin (ogen) exist in the blood circulation, such as anti-plasmin and the like. Normally, the activity of fibrinolytic substances in blood is many times higher than that of fibrinolytic substances, so fibrinolytic bleeding does not occur. However, these antagonists do not block the activation of activators (e.g., urokinase, etc.) that have been adsorbed onto the fibrin network to form plasmin. Plasmin is an endopeptidase that cleaves arginine and lysine peptide chains of fibrin (ogen) in a neutral environment to form fibrin degradation products and cause clot lysis bleeding. Plasminogen is specifically adsorbed on fibrin through lysine binding sites in its molecular structure, and lysine competitively inhibits this adsorption, reducing the adsorption rate of plasminogen, and thus reducing the activation of plasminogen to reduce bleeding. The product has similar spatial configuration to lysine (1, 5-diaminocaproic acid), and can competitively inhibit plasminogen from adsorbing on fibrin network, thereby preventing activation thereof, and protecting fibrin from degradation by plasmin to achieve hemostasis effect.
The effective blood concentration of the aminomethylbenzoic acid can be maintained for 3 to 5 hours after intravenous injection. After the medicine is taken for 24 hours, 36% +/-5% is excreted with urine in an original form, 63% +/-17% is excreted by intravenous injection, and the rest is acetylated derivatives. The gastrointestinal absorption rate after oral administration of aminomethylbenzoic acid was 69% and 2%. The distribution concentration in the body is kidney, liver, heart, spleen, lung, blood and the like in sequence. The peak value of the blood concentration can be reached 3 hours after the medicine is taken, the weight of the oral medicine is 7.5mg/kg, and the peak value is generally 4-5 ug/ml. The blood concentration is reduced to a very low level after 8 hours of oral administration; the effective blood concentration after intravenous injection may be in the range of 3 to 5 hours. After the medicine is taken for 24 hours, 36% +/-5% is excreted with urine in an original form, 63% +/-17% is excreted by intravenous injection, and the rest is acetylated derivatives.
The aminomethylbenzoic acid is prepared by a chemical synthesis method, for example, Chinese patent application No. 201810634724.X discloses a synthesis method of aminomethylbenzoic acid, which comprises the following specific steps: adding 4-halomethyl alkyl benzoate and triethylamine, dropwise adding an ethanol-water solution dissolved with 2-amino-5-methyl-1, 3, 4-thiadiazole while stirring, after complete reaction, evaporating the solution until a large amount of solids are separated out, cooling, filtering and drying to obtain 4-aminomethyl alkyl benzoate; adding 4-aminomethyl alkyl benzoate into an acid solution, stirring for reaction, cooling, adding water, dropwise adding alkali until the solution becomes alkaline and a large amount of solids are separated out, filtering, washing and drying to obtain aminomethylbenzoic acid. The method is mild in reaction condition, does not use-CN with strong toxicity and organic solvent with strong pollution, has the advantages of cheap and easily-obtained reaction raw materials, less side reactions, high yield, low cost, short reaction and post-treatment time, low energy consumption, high production efficiency, small environmental pollution and simple three-waste treatment, and is suitable for industrial production.
Chinese patent application No. 201510338012.X discloses a preparation method of aminomethylbenzoic acid. Specifically, the preparation method comprises the steps of taking p-cyanobenzyl halide (especially p-cyanobenzyl chloride) as a raw material, carrying out ammonolysis reaction under the catalysis of urotropine to obtain an intermediate p-cyanobenzylamine, and further carrying out acid hydrolysis to obtain a final product, namely the aminomethylbenzoic acid. The method is believed to have short synthesis steps, cheap and easily obtained raw materials, high yield, low cost, good quality, small pollution and simple and convenient operation, and is suitable for large-scale industrial production.
Chinese patent application No. 202010923736.1 relates to a method for preparing aminomethylbenzoic acid. The method comprises the following steps: p-methyl benzoic acid and a chlorinating agent are subjected to chlorination reaction under the condition of a catalyst I to obtain an intermediate p-chloromethyl benzoic acid, and then the intermediate p-chloromethyl benzoic acid and ammonia water are subjected to ammonolysis under the condition of a catalyst II to prepare the aminomethylbenzoic acid. The purity of the obtained aminomethylbenzoic acid product is more than 99.9%, the single impurity content is less than 0.1%, and the overall yield of the two-step reaction is more than 63%. The method is believed to have the advantages of short synthetic route, no use of highly toxic reagents or noble metals, low production cost, less environmental pollution and high overall yield, and is suitable for large-scale industrial production.
Chinese patent application No. 201110421970.5 relates to a new method for synthesizing aminomethylbenzoic acid bulk drug. The preparation method comprises the steps of using p-cyanobenzyl chloride as a raw material, performing acid hydrolysis to obtain an intermediate p-chloromethylbenzoic acid, and performing ammoniation synthesis under catalysis of urotropine to obtain aminomethylbenzoic acid; the method is believed to have short synthesis steps, cheap and easily obtained raw materials, high yield, low cost, good quality, little pollution and simple and convenient operation, and is very suitable for large-scale industrial production.
Chinese patent application No. 201810463637.2 discloses a synthesis method of aminomethylbenzoic acid, which is obtained by taking p-aldehyde benzoic acid as an initial raw material and performing catalytic hydrogenation amination; the temperature of the catalytic hydrogenation amination reaction is 80-150 ℃; the pressure of the catalytic hydrogenation amination reaction is 1.5-5.5 MPa; the amination reagent adopted by the catalytic hydrogenation amination is ammonia water or ammonia gas; the catalyst adopted by the catalytic hydrogenation amination is a palladium-carbon catalyst, and the dosage of the palladium-carbon catalyst is 1-10% of the weight of the p-aldehyde benzoic acid; after the catalytic hydrogenation amination, carrying out post-treatment to obtain an aminomethylbenzoic acid crude product and carrying out refining treatment to obtain an aminomethylbenzoic acid fine product; the refining treatment comprises alkali treatment, activated carbon decolorization and acid neutralization. The method is believed to have the advantages of short synthetic route, less environmental pollution, low production cost, no need of highly toxic articles such as sodium cyanide and liquid chlorine, high safety, high yield and suitability for industrial mass production.
Chinese patent application No. 201810677509.8 discloses a method for preparing aminomethylbenzoic acid, which belongs to the technical field of drug synthesis, and comprises the following steps: dissolving 4-chloromethyl benzoic acid in methanol to obtain a mixed solution; adding concentrated ammonia water into the mixed solution, controlling the temperature to be 40-80 ℃, reacting in a reaction container to convert 4-chloromethylbenzoic acid into 4-aminomethyl benzoic acid, and performing solid-liquid separation to obtain aminomethylbenzoic acid; the consumption of the ammonia water is greatly reduced, the recovery cost of the ammonia water is reduced, and the environmental pollution caused by the ammonia is reduced; the invention has low requirement on equipment, lower cost and simple production process, and can be popularized and applied in a large scale.
Chinese patent application No. 201910225125.7 discloses a method for preparing aminomethylbenzoic acid by recycling bromine, which comprises: carrying out bromination reaction on methyl benzoic acid and a bromination agent in the presence of a catalyst I and an oxidant to prepare p-bromomethyl benzoic acid, then carrying out ammonolysis on the p-bromomethyl benzoic acid in the presence of a catalyst II to obtain a product aminomethylbenzoic acid, and recovering bromine-containing mother liquor generated by the bromination reaction and the ammonolysis reaction by a method of precipitation, distillation and filtration. The yield of the product aminomethylbenzoic acid after recrystallization is not less than 96 percent, and the product purity is 99 percent. The brominating agent contains bromine element, and is selected from hydrogen bromide and acidic bromine-containing solution. The aminomethylbenzoic acid obtained by the method has high yield and purity, and can recycle bromine and ammonia water in a reaction system, thereby reducing the production cost.
Many of the methods for synthesizing aminomethylbenzoic acid described in the literature have a possibility that p-methylbenzoic acid as an impurity is introduced into the final product, and for example, the methods for synthesizing p-methylbenzoic acid directly from CN201910225125.7 and CN202010923736.1 are expected to be the above-mentioned methods, and the p-methylbenzoic acid as an impurity is occasionally detected in the conventional commercially available aminomethylbenzoic acid drug substances.
Chinese patent ZL201610502957.5 discloses a method for controlling the quality of a p-toluic acid bulk drug or an aminotoluic acid injection, which comprises the step of measuring the content of p-toluic acid and p-carboxychlorobenzyl chloride in a test sample by using a high performance liquid chromatography. In particular to a quality control method for measuring the content of p-toluic acid and p-carboxychlorobenzyl in a sample by using high performance liquid chromatography aiming at aminomethylbenzoic acid as a medicinal raw material drug and an injection containing the aminomethylbenzoic acid and water for injection.
The patent ZL201510150322.9 of the applicant discloses an aminomethylbenzoic acid freeze-dried powder injection pharmaceutical composition for injection, which comprises aminomethylbenzoic acid, a freeze-dried excipient such as mannitol and an optional acid-base regulator. In particular, the powder injection preparation contains a proper amount of dextran. The freeze-drying excipient is selected from sucrose, glucose, mannitol, lactose, sorbitol and glycine, and the weight ratio of the aminomethylbenzoic acid to the freeze-drying excipient is 100: 50 to 500. The aminomethylbenzoic acid freeze-dried powder injection for injection can be used for treating hemorrhage caused by primary fibrinolysis excess, including acute and chronic, localized or systemic high-fiber hemorrhage, which is commonly seen in cancer, leukemia, gynecological accidents, severe liver disease hemorrhage and the like, and has expected good properties. However, the invention does not give attention to the impurity p-toluic acid in the preparation, and it has been found that the quality detection method described in the prior art is not suitable for powder injections involved in ZL201510150322.9, especially for powder injections containing dextran therein.
Therefore, the skilled person still expects to provide clinically good pharmaceutical performance of aminomethylbenzoic acid preparations such as aminomethylbenzoic acid freeze-dried powder injection containing dextran, and especially to provide effective quality control method for such aminomethylbenzoic acid freeze-dried powder injection containing dextran, especially to detect impurity p-toluic acid in the aminomethylbenzoic acid freeze-dried powder injection containing dextran.
Disclosure of Invention
The invention aims to provide a freeze-dried powder injection containing aminomethylbenzoic acid with certain/certain good properties, which is expected to have one or more good pharmaceutical properties. Another aspect of the present invention is to provide a method for effectively controlling the quality of an aminomethylbenzoic acid freeze-dried powder injection containing dextran, and particularly a method for detecting p-toluic acid as an impurity in an aminomethylbenzoic acid freeze-dried powder injection containing dextran. The inventors have surprisingly found that freeze-dried powder injections comprising aminomethylbenzoic acid having a specific formulation have desirable good characteristics; the method can effectively detect the p-toluic acid which is an impurity in the powder injection. The present invention has been completed based on these findings.
In the present invention, the amount of aminomethylbenzoic acid as an active ingredient in the preparation of the composition is C8H9NO 2. H2O as p-aminomethylbenzoic acid monohydrate.
Accordingly, in a first aspect, the present invention provides a lyophilized powder for injection comprising aminomethylbenzoic acid and a lyophilized excipient.
The lyophilized powder injection according to any embodiment of the first aspect of the present invention, wherein the lyophilized excipient is selected from sucrose, glucose, mannitol, lactose, sorbitol, glycine.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, wherein the weight ratio of the aminomethylbenzoic acid to the freeze-dried excipient is 100: 50 to 500.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, wherein the weight ratio of the aminomethylbenzoic acid to the freeze-dried excipient is 100: 50 to 250.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, wherein the weight ratio of the aminomethylbenzoic acid to the freeze-dried excipient is 100: 50 to 200.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, wherein the weight ratio of the aminomethylbenzoic acid to the freeze-dried excipient is 100: 50 to 150.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, further comprises an acid-base regulator.
A lyophilized injectable powder according to any of the embodiments of the first aspect of the present invention, wherein the ph modifier is selected from sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, hydrochloric acid, phosphoric acid, nitric acid, sulfuric acid, or a combination thereof. In one embodiment, the pH adjusting agent is a hydrochloric acid solution or a sodium hydroxide solution, such as a 1M hydrochloric acid solution or a 1M sodium hydroxide solution.
The freeze-dried powder injection according to any one of the embodiments of the first aspect of the invention, wherein the amount of the pH regulator is such an amount that when the freeze-dried powder injection is dissolved into a solution containing aminomethylbenzoic acid at a concentration of 10mg/ml in terms of aminomethylbenzoic acid with water for injection, the pH of the solution is within a range of 3.0 to 5.0, for example, the pH of the solution is within a range of 3.5 to 4.5.
A lyophilized injectable powder according to any of the embodiments of the first aspect of the present invention further comprises dextran.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, wherein the dextran is selected from dextran-20, dextran-40 and dextran-70.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, wherein the weight ratio of the aminomethylbenzoic acid to the dextran is 100: 1 to 25.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, wherein the weight ratio of the aminomethylbenzoic acid to the dextran is 200: 2 to 20.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention comprises: 100 parts of aminomethylbenzoic acid, 50-200 parts of mannitol, 2-10 parts of dextran and an acid-base regulator; the dextran is selected from dextran-20, dextran-40 and dextran-70; the dosage of the acid-base regulator is such amount that when the freeze-dried powder injection is dissolved into a solution with the concentration of the aminomethylbenzoic acid of 10mg/ml by using water for injection, the pH value of the solution is within the range of 3.0-5.0; the acid-base regulator is hydrochloric acid solution or sodium hydroxide solution.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, wherein the weight ratio of aminomethylbenzoic acid to mannitol is 100: 50 to 150.
According to the freeze-dried powder injection of any embodiment of the first aspect of the invention, the pH regulator is 1M hydrochloric acid solution or 1M sodium hydroxide solution.
According to the freeze-dried powder injection of any embodiment of the first aspect of the invention, the dosage of the pH regulator is such an amount that when the freeze-dried powder injection is dissolved into a solution with the concentration of aminomethylbenzoic acid of 10mg/ml by using water for injection, the pH value of the solution is within the range of 3.5-4.5.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention is prepared by the following steps:
(a) weighing aminomethylbenzoic acid, mannitol and dextran according to the prescription amount, adding appropriate amount of water for injection for dissolving, adding active carbon, stirring, filtering and decarbonizing;
(b) adding water for injection to the prescription amount, stirring uniformly, measuring the pH value of the solution and optionally measuring the content of active ingredients, and adjusting the pH value to 3.0-5.0 by using an acid-base regulator according to the deviation condition of the pH value of the liquid medicine;
(c) sterilizing and filtering the liquid medicine, and filling the liquid medicine into a penicillin bottle;
(d) freeze drying to remove water, and pressing to obtain the final product,
wherein the proper amount of the water for injection in the step (a) is 70-90% of the prescription amount of the water for injection, and the prescription amount of the water for injection in the step (b) is 20-75 times of the weight of the aminomethylbenzoic acid.
The freeze-dried powder injection in any embodiment of the first aspect of the invention, wherein the dosage of the active carbon in the step (a) is 0.02-0.2% of the weight of the solution.
In the lyophilized powder for injection according to any one of the embodiments of the first aspect of the present invention, the "prescribed amount" of the "water for injection supplemented to the prescribed amount" in step (b) is 30 to 60 times the weight of aminomethylbenzoic acid.
According to the freeze-dried powder injection in any embodiment of the first aspect of the invention, the filtered filtrate obtained in the step (c) has a solid content of 2-15% (w/v).
According to the freeze-dried powder injection in any embodiment of the first aspect of the invention, the filtered filtrate obtained in the step (c) has a solid content of 2-10% (w/v).
According to the freeze-dried powder injection in any embodiment of the first aspect of the invention, the moisture content in the freeze-dried material obtained after removing the moisture in the step (d) is less than 5%.
As is well known, freeze-dried powder injection (usually, referred to as freeze-dried powder injection or freeze-dried powder injection) obtained by low-temperature freeze-vacuum drying is a powder or a block which is substantially anhydrous (usually, the water content is less than 5%, particularly, usually, less than 3%) and is obtained by dissolving each material in a solvent (usually, in water) to prepare a solution, freezing the solution at a low temperature, and then performing vacuum-pumping, sublimation and drying. Therefore, the pH of the solid lyophilizate is usually controlled by adjusting the pH of the solution during the formulation process; or can be controlled by adjusting the prescription so that the pH value of the obtained solid lyophilizate is controlled under the specified dissolving/diluting degree (the pH value of the solid lyophilizate is controlled); the latter method is generally more commonly used, for example, many lyophilized powder injections carried in pharmacopoeia control the pH value of the preparation in this way, and the pH value of the product can be controlled in this way, and the pH value of the final product can be only regulated without specifically specifying the prescription amount of the pH regulator. Also suitable for use in the present invention is the lyophilized powder for injection according to any embodiment of the first aspect of the present invention, wherein the optional amount of the pH adjusting agent is an amount such that when the prepared lyophilized powder for injection is dissolved with water for injection to form a solution containing aminomethylbenzoic acid at a concentration of 10mg/ml in terms of aminomethylbenzoic acid, the pH of the solution is within a range of 3.0 to 5.0, for example, the pH of the solution is within a range of 3.5 to 4.5.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention is prepared by the following steps:
(a) weighing aminomethylbenzoic acid, freeze-drying excipient and optional dextran, adding appropriate amount of water for injection, dissolving, adding activated carbon, stirring, filtering, and removing carbon;
(b) supplementing the water for injection to the prescription amount, uniformly stirring, measuring the pH value of the solution and the content of optional active ingredients, and adjusting the pH value to 3.0-5.0, preferably 3.5-4.5 by using an acid-base regulator according to the deviation condition of the pH value of the liquid medicine;
(c) sterilizing and filtering the liquid medicine, and filling the liquid medicine into a penicillin bottle;
(d) freeze drying to remove water, and pressing.
The freeze-dried powder injection according to any one of the embodiments of the first aspect of the invention, wherein the appropriate amount of water for injection in step (a) is about 70-90% of the amount of water for injection prescribed.
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, wherein the amount of the activated carbon used in the step (a) is 0.02-0.5% (w/v), preferably 0.02-0.2% of the weight of the solution.
The lyophilized powder for injection according to any embodiment of the first aspect of the present invention, wherein the "prescribed amount" of water for injection in the "supplementing water for injection to its prescribed amount" in step (b) is 10 to 100 times, for example 20 to 75 times, for example about 30 to 60 times the weight of aminomethylbenzoic acid. The amount of this water for injection can be easily controlled by the solids content in step (c).
The freeze-dried powder injection according to any embodiment of the first aspect of the invention, wherein the pH modifying agent in step (b) is an aqueous solution of a pH modifying agent selected from the group consisting of: sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, hydrochloric acid, phosphoric acid, nitric acid, sulfuric acid, or a combination thereof. The concentration of these aqueous solutions is well known to those skilled in the art, for example 1 to 10%, for example 2 to 5%. In one embodiment, the pH adjusting agent is a hydrochloric acid solution or a sodium hydroxide solution, such as a 1M hydrochloric acid solution or a 1M sodium hydroxide solution.
The freeze-dried powder injection according to any one of the embodiments of the first aspect of the present invention, wherein the filtered filtrate obtained in step (c) has a solid content of 2-20% (w/v), preferably 2-15% (w/v), and more preferably 2-10%.
The lyophilized powder injection according to any embodiment of the first aspect of the present invention, wherein the moisture content of the freeze-dried material obtained after removing the moisture in step (d) is less than 10%, preferably less than 8%, preferably less than 5%, more preferably less than 3%.
Further, the second aspect of the present invention provides a method for preparing a lyophilized powder for injection according to any embodiment of the first aspect of the present invention, which substantially comprises the following steps:
(a) weighing aminomethylbenzoic acid, freeze-drying excipient and optional dextran, adding appropriate amount of water for injection, dissolving, adding activated carbon, stirring, filtering, and removing carbon;
(b) supplementing the water for injection to the prescription amount, uniformly stirring, measuring the pH value of the solution and the content of optional active ingredients, and adjusting the pH value to 3.0-5.0, preferably 3.5-4.5 by using an acid-base regulator according to the deviation condition of the pH value of the liquid medicine;
(c) sterilizing and filtering the liquid medicine, and filling the liquid medicine into a penicillin bottle;
(d) freeze drying to remove water, and pressing.
The method according to any embodiment of the second aspect of the present invention, wherein the appropriate amount of water for injection in step (a) is about 70-90% of the amount of water for injection prescribed.
The process according to any one of the embodiments of the second aspect of the present invention, wherein the amount of activated carbon used in step (a) is 0.02% to 0.5% (w/v), preferably 0.02% to 0.2% by weight of the solution.
The method according to any one of the embodiments of the second aspect of the present invention, wherein the "prescribed amount" of water for injection in the "supplementing water for injection to its prescribed amount" in step (b) is 10 to 100 times, for example 20 to 75 times, for example about 30 to 60 times the weight of aminomethylbenzoic acid. The amount of this water for injection can be easily controlled by the solids content in step (c).
The method according to any of the embodiments of the second aspect of the present invention, wherein the pH modifying agent in step (b) is an aqueous solution of a pH modifying agent selected from the group consisting of: sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, hydrochloric acid, phosphoric acid, nitric acid, sulfuric acid, or a combination thereof. The concentration of these aqueous solutions is well known to those skilled in the art, for example 1 to 10%, for example 2 to 5%. In one embodiment, the pH adjusting agent is a hydrochloric acid solution or a sodium hydroxide solution, such as a 1M hydrochloric acid solution or a 1M sodium hydroxide solution.
The process according to any embodiment of the second aspect of the present invention, wherein the filtered filtrate obtained in step (c) has a solids content of 2-20% (w/v), preferably 2-15% (w/v), more preferably 2-10%.
The process according to any embodiment of the second aspect of the present invention, wherein the freeze-dried material obtained after removal of water in step (d) has a water content of less than 10%, preferably less than 8%, preferably less than 5%, more preferably less than 3%.
Further, the third aspect of the present invention provides a method for quality detection of the freeze-dried powder injection of any one of the first aspect of the present invention, in particular a method for quality detection of a freeze-dried powder injection containing dextran, which includes an operation of determining the content of p-toluic acid (which is also referred to as impurity X in the present invention) as an impurity in the freeze-dried powder injection by using high performance liquid chromatography.
The method according to the third aspect of the invention, comprising the operations of:
performing measurement according to the specification of high performance liquid chromatography recorded in section 0512 of the general rules of four departments in 2020 edition of Chinese pharmacopoeia;
preparation of phosphate buffer solution: dissolving 10.0-15.0 g (such as 12.0 g) of disodium hydrogen phosphate and 0.5-1 g (such as 0.8 g) of sodium heptanesulfonate in 900ml of water, adjusting the pH value to 2.0-3.0 (such as 2.5) by using phosphoric acid, adding water to 1000ml, and shaking up to obtain a phosphate buffer solution;
preparing a test solution: precisely weighing a sample containing 200mg of aminomethylbenzoic acid, adding 10ml of acetonitrile, 10-20 mg of ammonium chloride such as 15mg and 1-5 mg of ethylenediamine such as 3mg, mixing uniformly, performing ultrasonic treatment for 3min, filtering to obtain 2ml of subsequent filtrate, adding 8ml of phosphate buffer solution, mixing uniformly, precisely weighing a proper amount of mobile phase, and diluting to obtain a solution containing 400 mu g of aminomethylbenzoic acid in each 1ml, wherein the solution is used as a sample solution;
preparation of a control solution: precisely measuring a proper amount of a test solution, and quantitatively diluting with a mobile phase to prepare a solution containing 0.2-1.0 μ g, such as 0.4 μ g, of aminomethylbenzoic acid in 1ml of the solution as a control solution;
preparation of system applicability solution: taking a proper amount of aminomethylbenzoic acid and an impurity X reference substance, adding a mobile phase for dissolving and diluting to prepare solutions containing 400 mu g and 10 mu g of aminomethylbenzoic acid and impurity X in each 1ml as system applicability solutions respectively;
sensitivity solution: precisely measuring a proper amount of the control solution, and quantitatively diluting with a mobile phase to prepare a solution containing 0.1 mu g of aminomethylbenzoic acid in 1ml as a sensitive solution;
chromatographic conditions are as follows: a chromatographic column using octadecylsilane chemically bonded silica as a filler (the length of the chromatographic column is 200-300 mm, such as 250mm, the inner diameter of the chromatographic column is 4.6mm, and the particle size of the filler is 5 μm); phosphate buffer-acetonitrile (80: 20) is used as a mobile phase, and the flow rate is 1 ml/min; the column temperature is 40 ℃; the detection wavelength is 230 nm; the injection volume is 20 mul;
system applicability requirements: injecting the system applicability solution into a chromatographic column, recording a chromatogram, wherein the number of theoretical plates is not less than 5000 calculated according to an aminomethylbenzoic acid peak, and the separation degree between the aminomethylbenzoic acid peak and an impurity X peak is not less than 5.0; in the sensitivity solution chromatogram, the signal-to-noise ratio of the peak height of the main component is not less than 10;
the determination method comprises the following steps: precisely measuring a test solution, a reference solution and a sensitivity solution, respectively injecting into a liquid chromatograph, and recording the chromatogram until the retention time of a main peak is 4 times;
calculating the content of the impurity X: and calculating the content of the impurity X relative to the aminomethylbenzoic acid in the chromatogram of the test solution by taking the peak area of the main peak of the control solution as a control.
In the above-described steps of the preparation method of the present invention, although the specific steps described therein are distinguished in some detail or in language description from the steps described in the preparation examples of the detailed embodiments below, those skilled in the art can fully summarize the above-described method steps in light of the detailed disclosure throughout the present disclosure.
Any embodiment of any aspect of the invention may be combined with other embodiments, as long as they do not contradict. Furthermore, in any embodiment of any aspect of the invention, any feature may be applicable to that feature in other embodiments, so long as they do not contradict.
The invention is further described below.
All documents cited herein are incorporated by reference in their entirety and to the extent such documents do not conform to the meaning of the present invention, the present invention shall control. Further, the various terms and phrases used herein have the ordinary meaning as is known to those skilled in the art, and even though such terms and phrases are intended to be described or explained in greater detail herein, reference is made to the term and phrase as being inconsistent with the known meaning and meaning as is accorded to such meaning throughout this disclosure.
The freeze-dried powder injection comprises the following active ingredients:
according to the present invention, the term "excipient" may also be referred to as adjuvants, fillers, etc.
As used herein, "pharmaceutically acceptable excipient" refers to an excipient that can be used to formulate a drug that has substantially no adverse effect on an organism and is generally tolerated by an organism.
In the present invention, the preferable lyophilized powder injection of the present invention is prepared into a solution containing 100mg of the active ingredient in aminomethylbenzoic acid per 1ml with water, and then measured according to the method, i.e., pH measurement method, in the section VI H of the second part of the 2010 edition of the chinese pharmacopoeia.
Although it is understood by those skilled in the art that the excipient of the present invention may be any excipient that can be used for lyophilization, particularly mannitol, lactose, sucrose, glucose, sorbitol, glycine, and combinations thereof, in the present invention, especially preferred excipients are mannitol, lactose, and the like.
The preparation process of freeze-dried powder injection is a well-known pharmaceutical technology, such as two schematic freeze-drying curves, namely a freeze-drying curve A and a freeze-drying curve B, which are described in ZL 201510150322.9. In the following specific examples in the preparation of freeze-dried powder injections, the lyophilization profile used is lyophilization profile a, unless otherwise specified.
The water content in the freeze-dried powder injection is generally below 8%, preferably below 5%, more preferably below 3%. Moisture control can be controlled by appropriate adjustment of the freeze-drying program. The moisture content of the freeze-dried powder injection can be determined according to a number of known methods, such as loss-on-drying.
In the present invention, in order to adjust the pH of the drug solution as necessary, an appropriate pH adjusting agent (also referred to as an acid-base adjusting agent in the present invention) may be added to the composition. Although the present inventors have conducted adjustment only with a strong acid or strong alkali solution having no buffering power, such as an aqueous sodium hydroxide solution and an aqueous hydrochloric acid solution, it is understood by those skilled in the art that if treatment with such a pH adjusting agent having no buffering power is sufficient for the pH requirement of the system, the pH adjusting agent having buffering power will more achieve the object of the present invention, and therefore these buffering agents can not only adjust the pH but also stabilize the pH. Thus, any of the pH adjusting agents listed herein or combinations thereof are included within the spirit and scope of the present invention.
When the freeze-dried powder injection is prepared, the content of solid matters in the prepared liquid medicine is 2-20% (w/v), preferably 2-15% (w/v), and more preferably 2-10%. Since lyophilized injectable powder is usually obtained by lyophilization in a vial, it is understood by those skilled in the art that the product usually takes the shape of a cake before the final product is obtained even before it is used by a doctor, although the volume of the cake is theoretically smaller (slightly smaller) than the volume of the original aqueous solution, usually the volume of the cake is not reduced to 50% of the volume of the original aqueous solution, usually the volume of the cake is between 80-120% of the volume of the original aqueous solution, more usually the volume of the cake is between 90-100% of the volume of the original aqueous solution, the volume of the aqueous solution of the freeze-dried composition before freeze-drying can be estimated from the liquid level trace of the raw aqueous solution (the liquid level trace remained on the wall of the vial after the main cake is reduced by freeze-drying, and even if the freeze-dried product in the vial is powdery due to various reasons such as collision, the original liquid level trace can be usually remained). Therefore, although the invention provides a substantially anhydrous freeze-dried powder injection, the volume of the liquid medicine at least before the start of freeze-drying can be roughly estimated according to the powder injection, and the content of the solid in the prepared liquid medicine can be calculated according to the estimated volume and the weight of the dried final product in a penicillin bottle when the freeze-dried powder injection is prepared. Therefore, the solid content of the liquid medicine in the freeze-dried powder injection according to the first aspect of the present invention is 2-20% (w/v), preferably 2-15% (w/v), and even more preferably 2-10%.
The term "solid content" means that the solid substance (e.g. the active compound of the invention and all excipients used, weight/g) is added to a solvent (e.g. water for injection) and dissolved to give a solution, the weight of the solid substance being divided by the percentage of the final solution volume (weight/volume percentage, e.g. g/100 ml). For example, in the present invention, if a solution with a final volume of 100ml is prepared by adding an appropriate amount of an aqueous solution for injection to 5g of the total amount of the raw material drug and all the solid excipients, the solid content of the liquid drug is 5%.
In the present invention, the symbol%, depending on the context in which it is used, may have a meaning that is readily understood by a person skilled in the art. For example, where reference is made to solids content, the symbol indicates weight/volume percent (w/v, e.g., g/100 ml); also for example, where reference is made to "water content" in a freeze-dried powder injection, for example, the water content is below 8%, then the symbol% indicates weight/weight percent (w/w, g/100 g). Generally,% means weight/volume percent when a solid is dispersed in a liquid; where a solid is dispersed in a solid or a liquid is dispersed in a solid (e.g., the moisture content of a powder injection),% means weight/weight percent. In other cases, the symbol% represents weight/weight percent, unless otherwise indicated.
In preparing the drug solution of the present invention, it is known to those skilled in the art that rough filtration may be performed using, for example, a microfiltration membrane of about 0.45um, and fine filtration may be performed using, for example, a microfiltration membrane of about 0.22um to sterilize before filling the drug solution into vial, and that filtration may be performed several times if necessary.
The freeze-dried powder injection is freeze-dried powder injection. In one embodiment, the lyophilized powder for injection is a single dose formulation (e.g., vial-loaded powder for injection) and the amount of active compound per unit dose (which, as not otherwise specified in this disclosure, is converted to aminomethylbenzoic acid) may be, for example, but not limited to, about 20mg, about 50mg, about 100mg, about 150mg, about 200 mg.
The freeze-dried powder injection is dissolved again by water for injection, and the dissolving time is usually within 60 seconds, preferably within 50 seconds, and more preferably within 40 seconds.
According to the freeze-dried powder injection, after the solution containing 100mg of aminomethylbenzoic acid as an active ingredient in every 1ml of water is prepared, the pH value of the solution is 3.0-5.0. In one embodiment, the pH is 3.5 to 4.5.
The freeze-dried powder injection provided by the invention can be stored in a dry place below 25 ℃ for at least 24 months, and can meet the storage requirements of common freeze-dried powder injection.
The freeze dried powder for injection, especially freeze dried powder for injection, is white or white-like freeze dried block or its crushed block or powder, and has no bad smell, bitter taste and easy water solubility.
Aminomethylbenzoic acid powder injection is mainly used for treating hemorrhage caused by primary fibrinolysis excess, including acute and chronic, localized or systemic high-fiber hemorrhage, which is commonly seen in cancer, leukemia, gynecological accidents, severe liver disease hemorrhage and the like. Generally speaking, the content of the active ingredient in each bottle of the aminomethylbenzoic acid freeze-dried powder injection is 0.1 g. The aminomethylbenzoic acid freeze-dried powder injection is generally injected or instilled by vein, 0.1-0.3 g (1 to 3) per time, and no more than 0.6g (6) per day; or following the medical advice. Patients were monitored for the possibility of thrombotic complications when using lyophilized powder for injection of aminomethylbenzoic acid. It should be used with cautions for patients with thrombophilia (such as acute myocardial infarction). The aminomethylbenzoic acid freeze-dried powder injection is not used for secondary fibrinolytic hemorrhage caused by disseminated intravascular coagulation in general to prevent further thrombosis and influence organ functions, especially acute renal failure. If necessary, the product should be applied on the basis of heparinization. If combined with other blood coagulation factors (such as factor IX), the patient should be alert to thrombosis. It is considered that it is better to use the product 8 hours after the blood coagulation factor is used. The aminomethylbenzoic acid freeze-dried powder injection can cause the secondary renal pelvis and ureter blood clot blockage, and is used with caution when the hemophilia or the renal pelvis parenchyma lesion generates a large amount of hematuria. The heparin treatment is safer than the product for hypofibrinogenemia hemorrhage caused by intrauterine fetal death. The dosage is reduced when chronic renal function is incomplete, and the urine concentration is often higher after administration. The dosage should be reduced when treating the bleeding caused by prostate operation. During the period of using the aminomethylbenzoic acid lyophilized powder injection, a doctor should be consulted if any adverse event and/or adverse reaction occurs.
Aminomethylbenzoic acid is an aminomethylbenzoic acid type antifibrinolytic agent. Has the same action mechanism as 6-amino acetic acid and tranexamic acid. The three-dimensional configuration of the fibrinolysin is similar to that of lysine, and the fibrinolysin can competitively inhibit the adsorption of the fibrinolysin and protect the fibrin from being degraded by the fibrinolysin, thereby achieving the hemostasis effect. The anti-fibrinolytic activity of the compound is 4 to 5 times stronger than that of 6-aminocaproic acid. Aminomethylbenzoic acid is generally used for hemorrhage caused by primary fibrinolysis in clinic, including acute or chronic, localized or systemic hyperfibrinolysis, which is commonly seen in cancer, leukemia, gynecological accidents, severe liver disease hemorrhage, etc. In the aspect of pharmacokinetics, the effective blood concentration of the aminomethylbenzoic acid can be maintained for 3 to 5 hours after intravenous injection. After the medicine is taken for 24 hours, 36% +/-5% is excreted with urine in an original form, 63% +/-17% is excreted by intravenous injection, and the rest is acetylated derivatives.
As mentioned above, the invention patent ZL201510150322.9 of the present applicant relates to a pharmaceutical composition of freeze-dried powder injection containing appropriate amount of dextran for injection, especially containing: 100 parts of aminomethylbenzoic acid, 50-200 parts of mannitol, 2-10 parts of dextran and an acid-base regulator; the dextran is selected from dextran-20, dextran-40 and dextran-70; the dosage of the acid-base regulator is such amount that when the freeze-dried powder injection is dissolved into a solution with the concentration of the aminomethylbenzoic acid of 10mg/ml by using water for injection, the pH value of the solution is within the range of 3.0-5.0; the acid-base regulator is hydrochloric acid solution or sodium hydroxide solution. The ZL201510150322.9 enables unknown RRT1.7 impurities carried in raw materials to be remarkably removed in a solution preparation process by adding dextran; the RRT1.7 impurity is detected by a periduckweed method (the periduckweed method is used for detecting the ammonia toluic acid injection and related substances thereof by an HPLC method, the journal of Chinese medicine industry, 2014, 45 (10): 956; in the invention, the HPLC method can be simply called the periduckweed method), and the periduckweed method is mainly used for detecting the p-chloromethylbenzoic acid which is a synthetic process intermediate of RRT1.4 in the ammonia toluic acid injection. Thus, for the dextran-containing aminomethylbenzoic acid powder injection preferred by the applicant's prior patent ZL201510150322.9, the RRT1.4 impurity and RRT1.7 impurity are both effective in monitoring or inhibiting the amount of the dextran-containing aminomethylbenzoic acid powder injection. However, it has been found that the duckweed method cannot be effectively used for detecting the impurity p-methylbenzoic acid in the powder injection, especially cannot detect the impurity p-methylbenzoic acid in the powder injection containing dextran, and the separation degree of the p-methylbenzoic acid from a main peak is about 1.62 when the duckweed method detects the p-methylbenzoic acid, so that the separation degree required by the common preparation impurity content determination is difficult to reach the requirement that the separation degree is more than 3.0.
It has been surprisingly found that the use of the HPLC method of the invention, which may be referred to as the HPLC-X method in the present invention, enables the efficient detection of the impurity p-toluic acid in the lyophilized powder injection comprising dextran to which the invention relates.
Detailed Description
The present invention will be further described by the following examples, however, the scope of the present invention is not limited to the following examples. It will be understood by those skilled in the art that various changes and modifications may be made to the invention without departing from the spirit and scope of the invention. The present invention has been described generally and/or specifically with respect to materials used in testing and testing methods. Although many materials and methods of operation are known in the art for the purpose of carrying out the invention, the invention is nevertheless described herein in as detail as possible. The following examples further illustrate the invention without limiting it. In the following examples, the pH adjuster (i.e., the acid-base adjuster in the present invention) is, as not otherwise described, a 1M sodium hydroxide solution or a 1M hydrochloric acid solution in such an amount that the pH of the solution prepared before freeze-drying is adjusted to a prescribed value or range determined by dissolving the water for injection for freeze-dried powder injection prepared to a solution containing 10mg/ml of aminomethylbenzoic acid, when the powder for injection is prepared. The following preparation steps are given for the purpose of illustration and are described in some detail based on the comparability of the examples, and the person skilled in the art can fully generalize the method for preparing a lyophilized powder injection according to the invention from the prior knowledge. In the following formulation various compositions were prepared, the total amount of formulation for each batch was 1000 bottles, as not otherwise specified, but when the formulation is specified, it was stated that 100mg of aminomethylbenzoic acid was contained per bottle.
In the following examples of the preparation of powder injection, the same batch of commercially available raw material drugs is used to prepare the preparation, unless otherwise specified. The detection of the bulk drugs and the powder injection according to the lemna giraldii method can effectively monitor the product quality of the preparation, such as the RRT1.7 impurity in the bulk drugs and the powder injection, as described in the prior invention patent ZL201510150322.9 of the applicant.
Powder injection preparation part
Preparation example 1 preparation of powder injection comprising aminomethylbenzoic acid
The formula is as follows:
| aminomethylbenzoic acid | 100mg， |
| Mannitol | 95mg， |
| Dextran-20 | 5mg， |
| pH regulator | To a pH of 4.0, and, |
| water for injection | Add appropriate amount to 4 ml. |
The preparation method comprises the following steps:
(1) weighing the main drug and the auxiliary material according to the prescription amount, placing the main drug and the auxiliary material into a stainless steel barrel, adding injection water with the prescription amount of about 80 percent to dissolve all the components, adding 0.1 percent (w/v) of activated carbon according to the volume of the solution, stirring for 30 minutes, filtering to remove the carbon, and adding the injection water to be close to the full amount of the prescription.
(2) The filtrate was sampled, the pH was measured, and if necessary, adjusted to a predetermined value with a pH adjuster (the predetermined value is the pH value measured by diluting the lyophilized powder with water for injection to a solution containing 10mg/ml of the active ingredient, the same applies hereinafter), and water for injection was further added to the full amount prescribed.
(3) The liquid medicine is firstly filtered by a 0.45um microporous filter membrane and then filtered for 2 times by a 0.22um microporous filter membrane.
(4) Filling each bottle with a liquid medicine containing 100mg of active ingredients into a penicillin bottle with a proper size (the liquid medicine accounts for about 1/3-1/4 of the full mouth volume of the penicillin bottle), and adding a rubber plug in half.
(5) Freeze-drying according to lyophilization curve a described herein to a moisture content of less than 3%; after the freeze drying is finished, hydraulic plugging is carried out; and (5) pricking an aluminum cover to obtain the aluminum alloy. The sample of preparation example 1 in the present invention may be abbreviated as Ex 1; samples from other preparations can be similarly represented.
Preparation example 2 preparation of powder injection containing aminomethylbenzoic acid
The formula is as follows:
the preparation method comprises the following steps:
(1) weighing the main drug and the auxiliary material according to the prescription amount, placing the main drug and the auxiliary material into a stainless steel barrel, adding injection water with the prescription amount of about 80 percent to dissolve all the components, adding 0.1 percent (w/v) of activated carbon according to the volume of the solution, stirring for 30 minutes, filtering to remove the carbon, and adding the injection water to be close to the full amount of the prescription.
(2) The filtrate was sampled, the pH was measured, and if necessary, adjusted to a predetermined value with a pH adjuster (the predetermined value is the pH value measured by diluting the lyophilized powder with water for injection to a solution containing 10mg/ml of the active ingredient, the same applies hereinafter), and water for injection was further added to the full amount prescribed.
(3) The liquid medicine is firstly filtered by a 0.45um microporous filter membrane and then filtered for 2 times by a 0.22um microporous filter membrane.
(4) Filling each bottle with a liquid medicine containing 100mg of active ingredients into a penicillin bottle with a proper size (the liquid medicine accounts for about 1/3-1/4 of the full mouth volume of the penicillin bottle), and adding a rubber plug in half.
(5) Freeze-drying according to lyophilization curve a described herein to a moisture content of less than 3%; after the freeze drying is finished, hydraulic plugging is carried out; and (5) pricking an aluminum cover to obtain the aluminum alloy. The sample of preparation example 1 in the present invention may be abbreviated as Ex 2; samples from other preparations can be similarly represented.
Preparation example 3 preparation of powder injection comprising aminomethylbenzoic acid
The formula is as follows:
| aminomethylbenzoic acid | 100mg， |
| Mannitol | 150mg， |
| Dextran-40 | 2mg， |
| pH regulator | To a pH of 3.5, and, |
| water for injection | Add appropriate amount to 5 ml. |
The preparation method comprises the following steps:
(1) weighing the main drug and the auxiliary material according to the prescription amount, placing the main drug and the auxiliary material into a stainless steel barrel, adding injection water with the prescription amount of about 80 percent to dissolve all the components, adding 0.1 percent (w/v) of activated carbon according to the volume of the solution, stirring for 30 minutes, filtering to remove the carbon, and adding the injection water to be close to the full amount of the prescription.
(2) The filtrate was sampled, the pH was measured, and if necessary, adjusted to a predetermined value with a pH adjuster (the predetermined value is the pH value measured by diluting the lyophilized powder with water for injection to a solution containing 10mg/ml of the active ingredient, the same applies hereinafter), and water for injection was further added to the full amount prescribed.
(3) The liquid medicine is firstly filtered by a 0.45um microporous filter membrane and then filtered for 2 times by a 0.22um microporous filter membrane.
(4) Filling each bottle with a liquid medicine containing 100mg of active ingredients into a penicillin bottle with a proper size (the liquid medicine accounts for about 1/3-1/4 of the full mouth volume of the penicillin bottle), and adding a rubber plug in half.
(5) Freeze-drying according to lyophilization curve a described herein to a moisture content of less than 3%; after the freeze drying is finished, hydraulic plugging is carried out; and (5) pricking an aluminum cover to obtain the aluminum alloy. The sample of preparation example 1 in the present invention may be abbreviated as Ex 3; samples from other preparations can be similarly represented.
Preparation example 4 preparation of powder injection comprising aminomethylbenzoic acid
The formula is as follows:
| aminomethylbenzoic acid | 100mg， |
| Mannitol | 200mg， |
| Dextran-70 | 5mg， |
| pH regulator | To a pH of 3.0, and, |
| water for injection | Add appropriate amount to 3 ml. |
The preparation method comprises the following steps:
(1) weighing main drugs and auxiliary materials according to the prescription amount, placing the main drugs and the auxiliary materials into a stainless steel barrel, adding injection water with the prescription amount of about 70% to dissolve all the components, adding 0.02% (w/v) of activated carbon according to the volume of the solution, stirring for 30 minutes, filtering to remove carbon, and adding the injection water to be close to the full amount of the prescription.
(2) The filtrate was sampled, the pH was measured, and if necessary, adjusted to a predetermined value with a pH adjuster (the predetermined value is the pH value measured by diluting the lyophilized powder with water for injection to a solution containing 10mg/ml of the active ingredient, the same applies hereinafter), and water for injection was further added to the full amount prescribed.
(3) The liquid medicine is firstly filtered by a 0.45um microporous filter membrane and then filtered for 2 times by a 0.22um microporous filter membrane.
(4) Filling each bottle with a liquid medicine containing 100mg of active ingredients into a penicillin bottle with a proper size (the liquid medicine accounts for about 1/3-1/4 of the full mouth volume of the penicillin bottle), and adding a rubber plug in half.
(5) Freeze-drying according to lyophilization curve a described herein to a moisture content of less than 3%; after the freeze drying is finished, hydraulic plugging is carried out; and (5) pricking an aluminum cover to obtain the aluminum alloy. The sample of preparation example 1 in the present invention may be abbreviated as Ex 4; samples from other preparations can be similarly represented.
Preparation example 5 preparation of powder injection comprising aminomethylbenzoic acid
The formula is as follows:
| aminomethylbenzoic acid | 100mg， |
| Mannitol | 75mg， |
| Dextran-20 | 5mg， |
| pH regulator | To a pH of 5.0, and, |
| water for injection | Add appropriate amount to 9 ml. |
The preparation method comprises the following steps:
(1) weighing main drugs and auxiliary materials according to the prescription amount, placing the main drugs and the auxiliary materials into a stainless steel barrel, adding injection water with the prescription amount of about 90 percent to dissolve all the components, adding 0.2 percent (w/v) of activated carbon according to the volume of the solution, stirring for 30 minutes, filtering to remove the carbon, and adding the injection water to be close to the full amount of the prescription.
(2) The filtrate was sampled, the pH was measured, and if necessary, adjusted to a predetermined value with a pH adjuster (the predetermined value is the pH value measured by diluting the lyophilized powder with water for injection to a solution containing 10mg/ml of the active ingredient, the same applies hereinafter), and water for injection was further added to the full amount prescribed.
(3) The liquid medicine is firstly filtered by a 0.45um microporous filter membrane and then filtered for 2 times by a 0.22um microporous filter membrane.
(4) Filling each bottle with a liquid medicine containing 100mg of active ingredients into a penicillin bottle with a proper size (the liquid medicine accounts for about 1/3-1/4 of the full mouth volume of the penicillin bottle), and adding a rubber plug in half.
(5) Freeze-drying according to lyophilization curve B described herein to a moisture content of less than 3%; after the freeze drying is finished, hydraulic plugging is carried out; and (5) pricking an aluminum cover to obtain the aluminum alloy. The sample of preparation example 1 in the present invention may be abbreviated as Ex 5; samples from other preparations can be similarly represented.
As recorded in ZL201510150322.9, the content of active ingredients of the powder injection prepared in the preparation examples 1 to 5, RRT1.7 impurity control, detection and the like all obtain good results.
Supplementary test A: the formulations and the preparation methods of the preparation examples 1 to 5 are referred to respectively, except that no dextran is added, and five batches of powder injection are prepared and are respectively referred to as preparation examples 1a to 5a or respectively abbreviated as Ex1a to Ex5 a.
Second, examine test example section
1. Performance investigation test example:
the test example examines the appearance and solubility of the powder injection. It has been found that all the powder injections obtained in the above preparation examples 1 to 5 have the abnormal appearance of firm, intact round cake, no split, no spray bottle, etc.
All batches of the powder injection obtained in preparation examples 1 to 5 were taken, the plastic top of the bottle cap was opened, water for injection (the amount of the solution was about 2 times of the volume of the solution before freeze-drying of the corresponding sample) was injected from the bottle stopper by a syringe, the redissolution time was recorded by a stopwatch, and each batch of the sample was tested 5 times and averaged. As a result, the reconstitution time of all seven batches of powder injection samples is within the range of 15-43 seconds, for example, the reconstitution time of Ex1 is 24 seconds, and the samples have relatively quick clinically acceptable reconstitution performance.
2. Investigation of impurity p-methylbenzoic acid in powder injection
P-methylbenzoic acid, CAS No.: 99-94-5, available from Sigma-Aldrich, cat # T36803, also referred to herein as impurity X.
The impurity p-methylbenzoic acid in the raw material medicines, powder injection and other materials is measured by using the following [ HPLC-X method ], wherein five powder injections from Ex1 to Ex5 and five powder injections from Ex1a to Ex5a are all prepared from the raw material medicines.
[ HPLC-X method ]:
performing measurement according to the specification of high performance liquid chromatography recorded in section 0512 of the general rules of four departments in 2020 edition of Chinese pharmacopoeia;
preparation of phosphate buffer solution: taking 12.0g of disodium hydrogen phosphate and 0.8g of sodium heptanesulfonate, adding 900ml of water for dissolving, adjusting the pH value to 2.5 by using phosphoric acid, adding water to 1000ml, and shaking up to be used as a phosphate buffer solution;
preparing a test solution: accurately weighing a sample (raw material or powder injection) containing 200mg of aminomethylbenzoic acid, adding 10ml of acetonitrile, 15mg of ammonium chloride and 3mg of ethylenediamine, uniformly mixing, carrying out ultrasonic treatment for 3min, filtering, taking 2ml of subsequent filtrate, adding 8ml of phosphate buffer solution, uniformly mixing, accurately weighing a proper amount of mobile phase, diluting to prepare a solution containing 400 mu g of aminomethylbenzoic acid in each 1ml, and taking the solution as a sample solution; [ acetonitrile 10ml, ammonium chloride 15mg, and ethylenediamine 3mg can be mixed in advance ]
Preparation of a control solution: precisely measuring a proper amount of a test solution, and quantitatively diluting with a mobile phase to prepare a solution containing 0.4 mu g of aminomethylbenzoic acid in 1ml of the solution as a control solution;
preparation of system applicability solution: taking a proper amount of aminomethylbenzoic acid and an impurity X reference substance, adding a mobile phase for dissolving and diluting to prepare solutions containing 400 mu g and 10 mu g of aminomethylbenzoic acid and impurity X in each 1ml as system applicability solutions respectively;
sensitivity solution: precisely measuring a proper amount of the control solution, and quantitatively diluting with a mobile phase to prepare a solution containing 0.1 mu g of aminomethylbenzoic acid in 1ml as a sensitive solution;
chromatographic conditions are as follows: a column using octadecylsilane bonded silica as a packing (the column used in this example had a column length of 250mm, a column inner diameter of 4.6mm, and a packing particle size of 5 μm); phosphate buffer-acetonitrile (80: 20) is used as a mobile phase, and the flow rate is 1 ml/min; the column temperature is 40 ℃; the detection wavelength is 230 nm; the injection volume is 20 mul;
system applicability requirements: injecting the system applicability solution into a chromatographic column, and recording a chromatogram (the retention time of an aminomethylbenzoic acid peak is about 9 minutes and the relative retention time of the impurity X is 1.27 when the chromatographic conditions are used), wherein the number of theoretical plates is not less than 5000 according to the aminomethylbenzoic acid peak, and the separation degree between the aminomethylbenzoic acid peak and the impurity X peak is not less than 5.0; in the sensitivity solution chromatogram, the signal-to-noise ratio of the peak height of the main component is not less than 10 (the parameters meet the requirements of general drug inspection specifications);
the determination method comprises the following steps: precisely measuring a test solution, a reference solution and a sensitivity solution, respectively injecting into a liquid chromatograph, and recording the chromatogram until the retention time of a main peak is 4 times;
calculating the content of the impurity X: and calculating the content of the impurity X relative to the aminomethylbenzoic acid in the chromatogram of the test solution by taking the peak area of the main peak of the control solution as a reference (namely, the content of the impurity X is 0.1% when the peak area of the impurity X in the chromatogram of the test solution is equal to the peak area of the main peak of the control solution, the content of the impurity X is 0.5% when the peak area of the impurity X in the chromatogram of the test solution is 5 times of the peak area of the main peak of the control solution, and the like).
Generally, it is acceptable to have a level of impurity X in the formulation of less than 0.5%, preferably less than 0.25%, more preferably less than 0.2%.
The HPLC-X method is used for determining the content of impurity X in aminomethylbenzoic acid bulk drugs, five powder injections from Ex1 to Ex5 and five powder injections from Ex1a to Ex5 a; as a result: the impurity X content of the aminomethylbenzoic acid bulk drug is 0.087 percent, the impurity X content of five powder injections from Ex1 to Ex5 is in the range of 0.083 to 0.095 percent, such as 0.086 percent of the impurity X content of Ex1 powder injections, and the impurity X content of five powder injections from Ex1a to Ex5a is in the range of 0.082 to 0.093 percent, such as 0.089 percent of the impurity X content of Ex1a powder injections; as the content of the impurity X determined by an HPLC-X method is relative to the content of the main component aminomethylbenzoic acid, the results show that the content of the impurity X in the raw material and the content of the impurity X in the preparation are consistent and can not be changed due to factors such as preparation process, auxiliary material addition and the like of different powder injections prepared from the same batch of raw materials.
Placing aminomethylbenzoic acid crude drug, five powder injections of Ex 1-Ex 5 and five powder injections of Ex1 a-Ex 5a at 40 + -2 deg.C for 6 months, and measuring impurity X content of the samples for 6 months by HPLC-X method; as a result: the impurity X content of the aminomethylbenzoic acid bulk drug is 0.088 percent, the impurity X content of five powder injections from Ex1 to Ex5 is in the range of 0.084 to 0.097 percent, such as 0.087 percent of the impurity X content of Ex1 powder injections, and the impurity X content of five powder injections from Ex1a to Ex5a is in the range of 0.085 to 0.094 percent, such as 0.088 percent of the impurity X content of Ex1a powder injections; these results show that the content of impurity X relative to aminomethylbenzoic acid is essentially unchanged after the various materials have been subjected to the stability test.
Referring to the HPLC-X method, but not adding ethylenediamine in the preparation process of a test solution, determining the content of impurity X in aminomethylbenzoic acid bulk drugs, five powder injections from Ex1 to Ex5 and five powder injections from Ex1a to Ex5 a; as a result: the impurity X content of the aminomethylbenzoic acid raw material drug is 0.089 percent, the impurity X content of five powder injections from Ex1 to Ex5 is in the range of 0.033 to 0.046 percent, such as 0.037 percent of the impurity X content of Ex1 powder injections, and the impurity X content of five powder injections from Ex1a to Ex5a is in the range of 0.084 to 0.094 percent, such as 0.090 percent of the impurity X content of Ex1a powder injections; due to the good stability of impurity X, no major changes should occur with respect to aminomethylbenzoic acid during the formulation preparation process, and it was surprisingly found that the content of impurity X in Ex 1-Ex 5 powder injections could not be accurately determined using the HPLC-X method variant (without addition of ethylenediamine), while impurity X in Ex1 a-Ex 5a powder injections could still be accurately determined.
Referring to the HPLC-X method, but no ammonium chloride is added in the preparation process of a test solution, determining the content of impurity X in aminomethylbenzoic acid bulk drugs, five powder injections from Ex1 to Ex5 and five powder injections from Ex1a to Ex5 a; as a result: the content of impurity X in the aminomethylbenzoic acid bulk drug is 0.087%, the content of impurity X in five powder injections from Ex1 to Ex5 is in the range of 0.026-0.035%, such as 0.031% of impurity X in Ex1 powder injections, and the content of impurity X in five powder injections from Ex1a to Ex5a is in the range of 0.082-0.093%, such as 0.088% of impurity X in Ex1a powder injections; it has been found that the content of impurity X in Ex1 to Ex5 powder injections cannot be accurately determined using the HPLC-X method variant (without adding ammonium chloride), while impurity X in Ex1a to Ex5a powder injections can still be accurately determined.
Referring to the HPLC-X method, but adding no ammonium chloride and no ethylenediamine in the preparation process of the test solution, determining the content of impurity X in aminomethylbenzoic acid bulk drug, five powder injections from Ex1 to Ex5 and five powder injections from Ex1a to Ex5 a; as a result: the impurity X content of the aminomethylbenzoic acid raw material drug is 0.089 percent, the impurity X content of five powder injections from Ex1 to Ex5 is in the range of 0.036-0.050 percent, such as 0.042 percent of the impurity X content of Ex1 powder injections, and the impurity X content of five powder injections from Ex1a to Ex5a is in the range of 0.084-0.095 percent, such as 0.090 percent of the impurity X content of Ex1a powder injections; it has been found that the impurity X content in Ex1 to Ex5 powder injections cannot be accurately determined using the HPLC-X method variant (without adding ammonium chloride and without adding ethylenediamine), while the impurity X in Ex1a to Ex5a powder injections can still be accurately determined.
From the above results, it was demonstrated that the simultaneous addition of both ammonium chloride and ethylenediamine in the HPLC-X method can improve the accuracy of the determination of impurity X, which was further verified by the following four recovery tests.
3. Recovery test by HPLC-X method
Recovery test a:
preparing an impurity X stock solution: precisely weighing a proper amount of impurity X reference substance, adding acetonitrile to dissolve the impurity X reference substance to prepare a solution with the concentration of about 4mg/ml, and taking the solution as an impurity X stock solution (prepared 4.016 mg/ml);
preparing a test solution: precisely weighing a sample (raw material or powder injection) which is equivalent to 200mg of aminomethylbenzoic acid, adding 0.5ml (or 1ml or 1.5ml) of impurity X stock solution, 9.5ml (or 9ml or 8.5ml, the total adding amount of the impurity X stock solution and acetonitrile is 10ml), 15mg of ammonium chloride and 3mg of ethylenediamine, uniformly mixing, carrying out ultrasonic treatment for 3min, filtering, taking 2ml of subsequent filtrate, adding 8ml of phosphate buffer solution, uniformly mixing, precisely weighing a proper amount of mobile phase to dilute into a solution containing 400 mu g of aminomethylbenzoic acid in each 1ml, and taking the solution as a sample solution additionally added with the impurity X; (obtaining a test sample solution in which 2.008mg, 4.016mg, or 6.024mg of impurity X was additionally added per 200mg of aminomethylbenzoic acid)
Preparation of a control solution: precisely measuring a proper amount of the test solution with the additional impurity X, and quantitatively diluting the solution by 1000 times by using a mobile phase to obtain a reference solution;
the rest operations are carried out according to the HPLC-X method, the recovery rate (in terms of percentage of components) of the impurity X after three amounts of the impurity X are added into each material is measured and calculated, and the average value is obtained by measuring each concentration of each sample for 3 times;
results for recovery of impurity X: the recovery rates of the low, medium and high three additive amounts of the aminomethylbenzoic acid raw material drug are respectively 99.6%, 100.4% and 99.8%, the recovery rates of the low, medium and high three additive amounts of five powder injections from Ex1 to Ex5 are respectively 99.2-100.7%, 99.5-100.6% and 99.3-100.2%, for example, the recovery rates of the low, medium and high three additive amounts of Ex1 powder injection are respectively 100.3%, 99.7% and 100.1%, the recovery rates of the low, medium and high three additive amounts of five powder injections from Ex1a to Ex5a are respectively 99.5-100.3%, 99.8-100.4% and 99.2-100.4%, for example, the recovery rates of the high, medium and high three additive amounts of Ex1a powder injection are respectively 99.8%, 100.2% and 99.;
these results indicate that the HPLC-X method has excellent recovery rate for determining impurity X in aminomethylbenzoic acid or two kinds of powder injection.
Recovery test B:
referring to the recovery rate test A, except that ethylenediamine is not added in the preparation process of the test solution, the recovery rate of the impurity X after three amounts of the impurity X are added to each material is determined and calculated;
results for recovery of impurity X: the recovery rates of the low, medium and high three additive amounts of the aminomethylbenzoic acid raw material drug are respectively 99.4%, 99.7% and 100.4%, the recovery rates of the low, medium and high three additive amounts of five powder injections from Ex1 to Ex5 are respectively in the ranges of 78.3-82.5%, 76.3-83.4% and 79.6-85.3%, for example, the recovery rates of the low, medium and high three additive amounts of Ex1 powder injection are respectively 80.8%, 79.1% and 82.3%, the recovery rates of the low, medium and high three additive amounts of five powder injections from Ex1a to Ex5a are respectively in the ranges of 99.2-100.4%, 99.5-100.7% and 99.4-100.2%, for example, the recovery rates of the high three additive amounts of Ex1a powder injection are respectively 100.2%, 99.8; these results indicate that the content of impurity X in Ex 1-Ex 5 powder injections could not be accurately determined using the HPLC-X method variant (without addition of ethylenediamine), the methodological recovery was low, and the recovery was substantially unaffected for Ex1 a-Ex 5a powder injections without dextran added to the formulation.
Recovery test C:
referring to the recovery rate test A, the recovery rate of the impurity X after adding three amounts of the impurity X to each material is determined and calculated except that no ammonium chloride is added in the preparation process of the test solution;
results for recovery of impurity X: the recovery rates of the low, medium and high three additive amounts of the aminomethylbenzoic acid raw material drug are respectively 99.7%, 100.1% and 99.5%, the recovery rates of the low, medium and high three additive amounts of five powder injections from Ex1 to Ex5 are respectively 65.4-72.7%, 63.8-70.3% and 66.2-71.5%, for example, the recovery rates of the low, medium and high three additive amounts of Ex1 powder injection are respectively 69.3%, 67.4% and 69.6%, and the recovery rates of the low, medium and high three additive amounts of five powder injections from Ex1a to Ex5a are respectively 99.6-100.7%, 99.4-100.5% and 99.7-100.5%, for example, the recovery rates of the high three additive amounts of Ex1a powder injection are respectively 99.8%, 99.5% and 99.9%; these results indicate that the content of impurity X in Ex 1-Ex 5 powder injections cannot be accurately determined using the HPLC-X method variant (without adding ammonium chloride), the methodological recovery is low, and the recovery for Ex1 a-Ex 5a powder injections without dextran added to the prescription is not substantially affected.
Recovery test D:
referring to the recovery test A, except that no ammonium chloride and no ethylenediamine are added in the preparation process of the test solution, the recovery of the impurity X after the addition of three amounts of the impurity X to each material is determined and calculated;
results for recovery of impurity X: the recovery rates of three low, medium and high addition amounts of aminomethylbenzoic acid raw material drugs are respectively 100.5%, 100.3% and 99.7%, the recovery rates of three low, medium and high addition amounts of five powder injections from Ex1 to Ex5 are respectively 74.1-78.3%, 72.6-80.4% and 75.7-79.2%, for example, the recovery rates of three low, medium and high addition amounts of Ex1 powder injections are respectively 75.2%, 77.7% and 76.4%, and the recovery rates of three low, medium and high addition amounts of five powder injections from Ex1a to Ex5a are respectively 99.3-100.5%, 99.6-100.2% and 99.6-100.4%, for example, the recovery rates of three high addition amounts of Ex1a powder injections are respectively 99.6%, 99.9% and 100.2%;
these results indicate that the impurity X content in Ex 1-Ex 5 powder injections could not be accurately determined using the HPLC-X method variant (without adding ammonium chloride and without adding ethylenediamine), the methodological recovery was low, and there was essentially no effect on the recovery of Ex1 a-Ex 5a powder injections without dextran added to the prescription.
According to the results, the ammonium chloride and the ethylenediamine added in the HPLC-X method are shown to be capable of obviously improving the recovery rate of the impurity X determination in the powder injection containing dextran, and furthermore, the invention surprisingly discovers that for the powder injection containing dextran with excellent stability, the ammonium chloride and the ethylenediamine are simultaneously added in the HPLC-X method of the invention to accurately determine the impurity X in the powder injection containing dextran, and the unexpected effect of the ammonium chloride and the ethylenediamine in the HPLC-X method of the invention is not expected at all in the prior art.
In the HPLC-X method and the recovery rate investigation thereof, additional experiments have found that, when preparing the phosphate buffer solution, the use amount of disodium hydrogen phosphate in the range of 10.0-15.0 g has no significant influence on the method, the use amount of sodium heptanesulfonate in the range of 0.5-1 g has no significant influence on the method, and it is preferable that the pH value of the phosphate buffer solution is adjusted to 2.5; when preparing a sample solution, it is preferable to add 15mg of ammonium chloride and 3mg of ethylenediamine; the concentration of the prepared control solution is within the range of 0.2-1.0 mug, so that the method is not influenced, and only simple adjustment is needed during calculation; the length of the chromatographic column is 200-300 mm, which has no obvious influence on the method, but the retention time can be pushed forward or moved backward, but has no influence on the method.
4. Quality detection example of powder injection:
for each of the powder injections obtained in the above preparation examples 1 to 5 and commercially available aminomethylbenzoic acid for injection (H20050339, produced by north gaku, containing 0.1g of aminomethylbenzoic acid per bottle), their acidity, clarity and color of the solution, related substances, loss on drying, sterility, pyrogen, visible foreign substances, insoluble particles, content and other quality/property indexes were examined by the following methods:
acidity: taking a bottle of a test article 1, adding 10ml of water to dissolve the test article, and determining according to a law (appendix VI H of the second part of the 2010 edition of Chinese pharmacopoeia), wherein the pH value is generally required to be 3.5-4.5 in the field.
Clarity and color of the solution: 5 bottles of the test article are taken, 5ml of water is respectively added for dissolving, and the solution is checked according to law (I IX B and IX A in the second division of the 2010 edition of the Chinese pharmacopoeia), and the solution is generally required to be clear and colorless in the field.
Related substances are as follows: precisely weighing appropriate amount of the contents under the different filling amount items, adding mobile phase for dissolving and diluting to prepare solution containing 0.16mg per 1ml as test solution; precisely measuring a proper amount of a test solution, adding a mobile phase for dilution, and preparing a solution containing 1.6 mu g of the test solution in each 1ml as a control solution; according to the chromatographic conditions under the content determination item, taking 20 mul of the reference solution, injecting the reference solution into a liquid chromatograph, adjusting the detection sensitivity to ensure that the peak height of the main component is 10-15% of the full range of the recorder, then respectively injecting 20 mul of the two solutions into the liquid chromatograph, and recording the chromatogram until the retention time of the peak of the main component is 2.5 times; measuring the sum of the areas of impurity peaks (except for auxiliary material peaks) in a chromatogram of a test solution, wherein the sum of the areas of the impurity peaks is usually required to be not more than 0.8 times (0.8%) of the area of a main peak of a control solution in the field; in addition, the substance measuring method of the present invention has been found to be unable to detect the impurity X, which is not separated from the main component and has a degree of separation of less than 1.2.
Loss on drying: a proper amount of a test sample is taken and dried to constant weight at 105 ℃ (the appendix VIII L of the second part of the 2010 version of Chinese pharmacopoeia), and the weight loss reduction amount is usually required to be not more than 3.5% in the field.
Pyrogen: taking a proper amount of a test sample, adding 0.9% sodium chloride solution for dissolving, preparing into a solution containing 10mg per 1ml, checking according to law (appendix XI D of the second part of the 2010 edition of Chinese pharmacopoeia) and injecting 5ml per 1kg of rabbit body weight according to the general requirement in the field, wherein the general requirement of the field is to meet the regulation.
And (3) sterilization: the test sample is dissolved in 0.9% sterile sodium chloride solution, and the solution is examined according to law (appendix XI H of the second part of the Chinese pharmacopoeia 2010), which generally requires the field to meet the requirements.
Visible foreign matter: the test article is taken and examined according to law (appendix IX H of the second part of the Chinese pharmacopoeia 2010 edition), and the general requirements in the field are in accordance with the regulations.
Insoluble microparticles: the test article is taken and examined according to law (appendix IX C of the second part of the Chinese pharmacopoeia 2010 edition), and the general requirements in the field are in accordance with the regulations.
Content determination: measuring by high performance liquid chromatography (appendix V D of second part of Chinese pharmacopoeia 2010 edition);
chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; taking 0.1mol/L ammonium acetate-glacial acetic acid (100: 1) as a mobile phase; the flow rate is 1.0 ml/min; the detection wavelength is 230 nm; the number of theoretical plates is not less than 3000 calculated according to aminomethylbenzoic acid peak;
the determination method comprises the following steps: taking contents under the items of different sample loading quantities, precisely weighing a proper amount (about equivalent to 32mg of aminomethylbenzoic acid) and placing the proper amount into a 100ml measuring flask, adding a mobile phase to dilute to a scale, shaking up, precisely weighing 1ml, placing the sample into a 10ml measuring flask, and adding the mobile phase to dilute to the scale to obtain a sample solution; accurately weighing appropriate amount of aminomethylbenzoic acid reference substance, adding mobile phase for dissolving and diluting to obtain solution containing 32 μ g of aminomethylbenzoic acid per 1ml, respectively taking 20 μ l of each solution, injecting into liquid chromatograph, recording chromatogram, and calculating according to external standard method by peak area to obtain the final product; it is generally required in the art that the aminomethylbenzoic acid (C8H9NO 2. H2O) should be 95.0% to 105.0% of the indicated amount (theoretical charge), calculated on the average charge.
The powder injection to be tested is detected according to the method, and all indexes are in the range generally specified by all the indexes.
5. Long-term stability test example:
regarding the five batches of the powder injections obtained in preparation examples 1 to 5 above and the commercially available aminomethylbenzoic acid for injection (H20050339, manufactured by North Kogaku, containing 0.1g of aminomethylbenzoic acid per bottle), these samples were allowed to stand at room temperature of 40. + -. 2 ℃ for 6 months, and the respective indices of these powder injections at 0 month and at 6 months were examined with reference to the method of "2, powder injection quality test example" above. The results show that after all six batches of powder injection are placed at 40 ℃ for 6 months, all detection indexes are still within the range generally specified by all the indexes. The powder injection of the invention is proved to have excellent stability.
6. Safety test example:
the test example is used for carrying out safety test investigation on the composition powder injection
The obtained samples Ex1 powder injection, Ex2 powder injection, Ex3 powder injection and commercially available powder injection (aminomethylbenzoic acid for injection, H20050339, produced by North Kogaku Huatai) are subjected to a blood vessel irritation experiment, a hemolysis experiment and an allergy experiment according to the requirements of the current drug registration regulations (research group of research on irritation, allergy and hemolysis research technology subject matter of chemical drug, the research technology guide principle of irritation, allergy and hemolysis of chemical drug, the research technology guide principle of drug, Beijing, China pharmaceutical science and technology Press, 2006: 124, and the results show that the samples all conform to the provisions of the blood vessel irritation experiment, the hemolysis experiment and the allergy experiment. The composition of the invention is shown to have good safety. For example, in the aspect of irritation, the skin and veins of the injection part of all the tested powder injection samples show similar changes, and abnormal phenomena such as congestion, edema, induration, necrosis and the like do not occur.
Claims (10)
1. The method for detecting the quality of the freeze-dried powder injection containing the aminomethylbenzoic acid and the freeze-dried excipient comprises the operation of measuring the content of the impurity p-toluic acid in the freeze-dried powder injection by using a high performance liquid chromatography.
2. The method according to claim 1, wherein in the lyophilized powder injection:
the freeze-drying excipient is selected from sucrose, glucose, mannitol, lactose, sorbitol and glycine;
the weight ratio of aminomethylbenzoic acid to lyophilized excipient is 100: 50-500;
the weight ratio of aminomethylbenzoic acid to lyophilized excipient is 100: 50-250 parts of;
the weight ratio of aminomethylbenzoic acid to lyophilized excipient is 100: 50-200 parts of; and/or
The weight ratio of aminomethylbenzoic acid to lyophilized excipient is 100: 50 to 150.
3. The method according to claim 1, wherein in the lyophilized powder injection:
also comprises an acid-base regulator;
the acid-base regulator is selected from sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, hydrochloric acid, phosphoric acid, nitric acid, sulfuric acid or the combination thereof;
the acid-base regulator is hydrochloric acid solution or sodium hydroxide solution, such as 1M hydrochloric acid solution or 1M sodium hydroxide solution; and/or
The dosage of the acid-base regulator is such amount that when the water for injection is dissolved into a solution containing the aminomethylbenzoic acid with the concentration of 10mg/ml calculated by the aminomethylbenzoic acid, the pH value of the solution is within the range of 3.0-5.0, for example, the pH value of the solution is within the range of 3.5-4.5.
4. The method according to claim 1, wherein in the lyophilized powder injection:
further comprising dextran;
the dextran is selected from dextran-20, dextran-40 and dextran-70;
the weight ratio of aminomethylbenzoic acid to dextran is 100: 1-25; and/or
The weight ratio of aminomethylbenzoic acid to dextran is 200: 2 to 20.
5. The method according to claim 1, wherein the lyophilized powder for injection comprises: 100 parts of aminomethylbenzoic acid, 50-200 parts of mannitol, 2-10 parts of dextran and an acid-base regulator; the dextran is selected from dextran-20, dextran-40 and dextran-70; the dosage of the acid-base regulator is such amount that when the freeze-dried powder injection is dissolved into a solution with the concentration of the aminomethylbenzoic acid of 10mg/ml by using water for injection, the pH value of the solution is within the range of 3.0-5.0; the acid-base regulator is hydrochloric acid solution or sodium hydroxide solution.
6. The method according to claim 1, wherein in the lyophilized powder injection:
the weight ratio of aminomethylbenzoic acid to mannitol is 100: 50-150 parts of;
the acid-base regulator is 1M hydrochloric acid solution or 1M sodium hydroxide solution; and/or
The dosage of the acid-base regulator is such amount that when the freeze-dried powder injection is dissolved into a solution with the concentration of the aminomethylbenzoic acid of 10mg/ml by using the water for injection, the pH value of the solution is within the range of 3.5-4.5.
7. The method according to claim 1, wherein the lyophilized powder for injection is prepared by the steps comprising:
(a) weighing aminomethylbenzoic acid, mannitol and dextran according to the prescription amount, adding appropriate amount of water for injection for dissolving, adding active carbon, stirring, filtering and decarbonizing;
(b) adding water for injection to the prescription amount, stirring uniformly, measuring the pH value of the solution and optionally measuring the content of active ingredients, and adjusting the pH value to 3.0-5.0 by using an acid-base regulator according to the deviation condition of the pH value of the liquid medicine;
(c) sterilizing and filtering the liquid medicine, and filling the liquid medicine into a penicillin bottle;
(d) freeze drying to remove water, and pressing to obtain the final product,
wherein the proper amount of the water for injection in the step (a) is 70-90% of the prescription amount of the water for injection, and the prescription amount of the water for injection in the step (b) is 20-75 times of the weight of the aminomethylbenzoic acid.
8. The method of claim 7, wherein:
the proper amount of the water for injection in the step (a) is about 70-90% of the prescription amount of the water for injection;
the dosage of the active carbon in the step (a) is 0.02 to 0.2 percent of the weight of the solution; and/or
The "prescribed amount" of water for injection in the "supplementing water for injection to its prescribed amount" in the step (b) is 30 to 60 times the weight of aminomethylbenzoic acid.
9. The method of claim 7, wherein:
the filtered filtrate obtained in the step (c) contains 2-15% (w/v) of solid;
the filtered filtrate obtained in the step (c) contains 2-10% (w/v) of solid; and/or
Removing water in step (d) to obtain a freeze-dried material having a water content of less than 5%.
10. A method according to claim 1, comprising the operations of:
performing measurement according to the specification of high performance liquid chromatography recorded in section 0512 of the general rules of four departments in 2020 edition of Chinese pharmacopoeia;
preparation of phosphate buffer solution: dissolving 10.0-15.0 g (such as 12.0 g) of disodium hydrogen phosphate and 0.5-1 g (such as 0.8 g) of sodium heptanesulfonate in 900ml of water, adjusting the pH value to 2.0-3.0 (such as 2.5) by using phosphoric acid, adding water to 1000ml, and shaking up to obtain a phosphate buffer solution;
preparing a test solution: precisely weighing a sample containing 200mg of aminomethylbenzoic acid, adding 10ml of acetonitrile, 10-20 mg of ammonium chloride such as 15mg and 1-5 mg of ethylenediamine such as 3mg, mixing uniformly, performing ultrasonic treatment for 3min, filtering to obtain 2ml of subsequent filtrate, adding 8ml of phosphate buffer solution, mixing uniformly, precisely weighing a proper amount of mobile phase, and diluting to obtain a solution containing 400 mu g of aminomethylbenzoic acid in each 1ml, wherein the solution is used as a sample solution;
preparation of a control solution: precisely measuring a proper amount of a test solution, and quantitatively diluting with a mobile phase to prepare a solution containing 0.2-1.0 μ g, such as 0.4 μ g, of aminomethylbenzoic acid in 1ml of the solution as a control solution;
preparation of system applicability solution: taking a proper amount of aminomethylbenzoic acid and an impurity X reference substance, adding a mobile phase for dissolving and diluting to prepare solutions containing 400 mu g and 10 mu g of aminomethylbenzoic acid and impurity X in each 1ml as system applicability solutions respectively;
sensitivity solution: precisely measuring a proper amount of the control solution, and quantitatively diluting with a mobile phase to prepare a solution containing 0.1 mu g of aminomethylbenzoic acid in 1ml as a sensitive solution;
chromatographic conditions are as follows: a chromatographic column using octadecylsilane chemically bonded silica as a filler (the length of the chromatographic column is 200-300 mm, such as 250mm, the inner diameter of the chromatographic column is 4.6mm, and the particle size of the filler is 5 μm); phosphate buffer-acetonitrile (80: 20) is used as a mobile phase, and the flow rate is 1 ml/min; the column temperature is 40 ℃; the detection wavelength is 230 nm; the injection volume is 20 mul;
system applicability requirements: injecting the system applicability solution into a chromatographic column, recording a chromatogram, wherein the number of theoretical plates is not less than 5000 calculated according to an aminomethylbenzoic acid peak, and the separation degree between the aminomethylbenzoic acid peak and an impurity X peak is not less than 5.0; in the sensitivity solution chromatogram, the signal-to-noise ratio of the peak height of the main component is not less than 10;
the determination method comprises the following steps: precisely measuring a test solution, a reference solution and a sensitivity solution, respectively injecting into a liquid chromatograph, and recording the chromatogram until the retention time of a main peak is 4 times;
calculating the content of the impurity X: and calculating the content of the impurity X relative to the aminomethylbenzoic acid in the chromatogram of the test solution by taking the peak area of the main peak of the control solution as a control.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117517534A (en) * | 2024-01-08 | 2024-02-06 | 吉林省长源药业有限公司 | Method for detecting maleic anhydride in succinic anhydride |
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| CN104198610A (en) * | 2014-09-16 | 2014-12-10 | 德州博诚制药有限公司 | Aminomethylbenzoic acid sample impurity detection device |
| CN104721153A (en) * | 2015-03-31 | 2015-06-24 | 山东北大高科华泰制药有限公司 | Aminomethylbenzoic acid freeze-dried powder injection medicine composition for injection |
| CN105929098A (en) * | 2016-06-30 | 2016-09-07 | 湖南洞庭药业股份有限公司 | Aminomethylbenzoic acid and injection composition and quality control method thereof |
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| CN104198610A (en) * | 2014-09-16 | 2014-12-10 | 德州博诚制药有限公司 | Aminomethylbenzoic acid sample impurity detection device |
| CN104721153A (en) * | 2015-03-31 | 2015-06-24 | 山东北大高科华泰制药有限公司 | Aminomethylbenzoic acid freeze-dried powder injection medicine composition for injection |
| CN105929098A (en) * | 2016-06-30 | 2016-09-07 | 湖南洞庭药业股份有限公司 | Aminomethylbenzoic acid and injection composition and quality control method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117517534A (en) * | 2024-01-08 | 2024-02-06 | 吉林省长源药业有限公司 | Method for detecting maleic anhydride in succinic anhydride |
| CN117517534B (en) * | 2024-01-08 | 2024-03-26 | 吉林省长源药业有限公司 | Method for detecting maleic anhydride in succinic anhydride |
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