CN108938626B - Carbazochrome sodium sulfonate pharmaceutical composition with good stability and high safety as well as preparation method and application thereof - Google Patents
Carbazochrome sodium sulfonate pharmaceutical composition with good stability and high safety as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a pharmaceutical composition with good stability and high safety, a preparation method and application thereof, wherein the pharmaceutical composition contains carbazochrome sodium sulfonate and a structural compound shown in a formula III with the content of not more than 0.5%.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to a carbazochrome sodium sulfonate pharmaceutical composition with good stability and high safety, and a preparation method and application thereof.
Background
As known in the art, physiological hemostatic mechanisms include: coagulation factor, platelet factor, vascular factor and fibrinolytic factor, 4 factors are closely related, and any deficiency thereof causes bleeding. According to the clinical classification of commonly used hemostatics, they can be classified into procoagulant drugs, antifibrolytic drugs and vasoactive drugs.
In the first category, the procoagulant drugs mainly comprise etamsylate, thrombin, vitamin K1 and the like. The second group, antifibrinolytics, are mainly tranexamic acid, ethylenediamine diacetoacetate, aminomethylbenzoic acid, aminocaproic acid, and the like. Third, the hemostatic acting on blood vessels mainly includes adrenal color hydrazone (carba-clo) and carbazochrome sodium sulfonate. The adrenal color hydrazone tablets and the carbazochrome sodium sulfonate injection are mainly used for reducing the permeability of capillary vessels, promoting the retraction of damaged capillary vessel ends to stop bleeding, and are mainly used for clinically treating bleeding caused by the increase of the permeability of capillary vessels such as small blood vessel rupture bleeding, hemoptysis, nosebleed, retinal blood and the like.
Adrenaline (Epinephrine) is a classic drug with strong anti-shock vasoactivity, acts by contracting blood vessels, and can be clinically used for local hemostasis in addition to rescue of anaphylactic shock, cardiac arrest, and the like. However, epinephrine is unstable and is easily oxidized to adrenaline (Adrenochrome), and it is found by Derouaux and Roskam, etc., which are university of Bili, though it still has a problem of instability. The adrenal color hydrazone (Adrenochlorobenzene, Adrenoxyl, Carbazochrome, Adrenol) modified from the chemical structure of epinephrine is commercialized due to the improvement of stability, is suitable for bleeding caused by capillary injury and permeability increase, such as epistaxis, retinal hemorrhage, hemoptysis, gastrointestinal hemorrhage, hematuria, hemorrhoids, uterine bleeding and the like, is also used for thrombocytopenic purpura, can be orally taken and injected, but can only be injected intramuscularly when being used for injection.
Until the date, Honda pharmaceutical corporation further modified the chemical structure of adrenal color hydrazone into sodium sulfonate salt
Carbazochrome sodium sulfonate injection (ADONA, AC-17) with various specifications is developed and marketed in the year, and can be used for intravenous injection only because the water solubility is further improved. The marketed specification of the carbazochrome sodium sulfonate injection is 2 ml: 10mg (subcutaneous or intramuscular), 5 ml: 25mg, 10 ml: 50mg and 20 ml: 100mg (the latter three are all used intravenously), and to date, oral carbazochrome sodium sulfonate tablets (10mg, 30mg), powders (100 mg: 1g) and fine granules (100 mg: 1g) are marketed in Japan, and the daily dose is
Is administered in three times. Different from the domestic standard calculation, the carbazochrome sodium sulfonate in the original research is calculated by hydrate, and the carbazochrome sodium sulfonate hydrate of 100mg is about equal to 85.6 on the basis of anhydrous substance in Chinamg of anhydrate.
Chemical name of carbazochrome sodium sulfonate: 1-methyl-6-oxo-2, 3, 5, 6-tetrahydroindol-5-semicarbazone-2-sulfonic acid sodium salt trihydrate having the formula C10H11N4NaO5S·3H2O, the structural formula of which is shown in formula I:
carbazochrome sodium sulfonate is mainly used for the following diseases: for bleeding tendency caused by decrease of capillary resistance and increase of permeability (e.g., purpura, etc.); fundus, renal, and uterine bleeding for exudation from the skin or mucosa and endometrium due to attenuation of capillary resistance; for intraoperative and postoperative abnormal bleeding caused by attenuation of capillary resistance. The Chinese medicine is mainly used for treating bleeding diseases of urinary system, upper digestive tract, respiratory tract and obstetrics and gynecology department in China. Has remarkable curative effect on urinary system, and can also be used for preventing and treating surgical hemorrhage, etc.
In order to improve the quality of carbazochrome sodium sulfonate and reduce the side effects thereof, researchers at home and abroad have made many studies, for example, japanese pharmacopoeia stipulates that the content of carbazochrome sodium sulfonate is not less than 98.0% and not more than 102.0%, and the content of related substances other than adrenal color hydrazone (carbazochrome, hereinafter, the same, the structural formula is shown as formula ii) is not more than 1.0% (high performance liquid chromatography, 360 nm). The content of carbazochrome sodium sulfonate is not less than 98.5%, the content of adrenal color hydrazone is not more than 2.0%, and the content of other related substances is not more than 1.0% (high performance liquid chromatography, 363 nm).
In order to better control the quality of carbazochrome sodium sulfonate and improve the safety, effectiveness and stability of carbazochrome sodium sulfonate, intensive research is necessary.
Disclosure of Invention
The inventor unexpectedly finds that a new chromatographic peak is detected at a wavelength lower than 360nm or 363nm when the high performance liquid chromatography is adopted to carry out full-wavelength scanning on carbazochrome sodium sulfonate, and further discovers a new compound with a structure shown in a formula III through further separation and purification.
Experiments show that the carbazochrome sodium sulfonate is unstable and is easy to be oxidized, degraded and the like, and the compound of the formula III is gradually increased along with the prolonging of the standing time of the carbazochrome sodium sulfonate and the like. Therefore, the purpose of improving the stability and safety of the carbazochrome sodium sulfonate medicament is realized by controlling the content of the compound with the structure shown in the formula III.
The invention aims to provide a carbazochrome sodium sulfonate pharmaceutical composition, which contains carbazochrome sodium sulfonate and a compound (also called a compound of formula III) with a structure shown in formula III, wherein the content of the compound is not higher than 1%.
Further, the carbazochrome sodium sulfonate in the pharmaceutical composition is not less than 90%, preferably not less than 95%, and more preferably not less than 98%.
Further, the content of the structural compound represented by the formula III is preferably not more than 1%, preferably not more than 0.75%, more preferably not more than 0.5%.
Further, the weight ratio of carbazochrome sodium sulfonate to the structural compound shown in formula III is not less than 1000:10, preferably not less than 1000:7.5, and more preferably not less than 1000: 5.
Further, the pharmaceutical composition contains carbazochrome sodium sulfonate with the content of not less than 98%, a structural compound shown in a formula III with the content of not more than 0.5%, adrenal color hydrazone with the content of not more than 1.0%, and the balance of other related substances.
Further, the content of adrenal color hydrazone in the medicine composition is not higher than 0.5%.
Further, the compound of the formula III is prepared by heating carbazochrome sodium sulfonate and alkali for destructive reaction, and can be detected by a high performance liquid chromatography method at a specific wavelength.
Further, the reaction occurs by placing the carbazochrome sodium sulfonate and the base in a solvent.
Further, the solvent for preparing the compound of formula III is selected from any one of methanol, ethanol, isopropanol, n-propanol, n-butanol and water or the combination thereof.
Further, the base for preparing the compound of the formula III is any one or combination of triethylamine, ammonia water, pyridine, sodium hydroxide, potassium hydroxide and disodium hydrogen phosphate.
Further, the preparation method of the compound of the formula III comprises heating for destroying, separating and purifying, and preferably, the purification method is selected from one or a combination of column chromatography and preparative liquid phase separation refining.
Further, the liquid phase separation refining condition for preparing the compound of the formula III is that the mobile phase is any one or the combination of methanol-water, acetonitrile-water and methanol-acetonitrile-water, isocratic or gradient elution is carried out, the product fractions are collected and evaporated to dryness or freeze drying is carried out, thus obtaining the compound of the formula III.
Further, the mobile phase of the liquid chromatography method for detecting the compound of the formula III is any one of or a combination of a methanol-salt buffer solution, an acetonitrile-salt buffer solution and a methanol-acetonitrile-salt buffer solution, and is subjected to isocratic or gradient separation, wherein the salt buffer solution is prepared from any one of or a combination of phosphoric acid, triethylamine, sodium acetate, glacial acetic acid and ammonium dihydrogen phosphate and water.
Further, the detection wavelength of the liquid chromatography method for detecting the compound of the formula III is 296nm, 280nm, 220nm, preferably 220 nm.
By adopting the separation and purification method of the invention, the content of the compound shown in the formula III is not less than 90%, preferably not less than 95%, and more preferably not less than 98%.
The invention aims to provide a preparation method of the carbazochrome sodium sulfonate (composition), which mainly comprises the following steps: adding sodium bisulfite solution into adrenal color hydrazone under nitrogen protection, heating and stirring until reaction is complete, filtering, concentrating the filtrate, refrigerating, precipitating crystals, filtering, washing with acetone, and drying to obtain the final product.
For the purposes of clearly presenting the context of the invention, the invention is defined by the following terms:
the "related substance" in the present invention refers to an impurity other than carbazochrome sodium sulfonate, such as adrenal color hydrazone, the compound of formula iii, and other substances contained in carbazochrome sodium sulfonate, which are not detected, and the "other related substance" refers to other substances contained in carbazochrome sodium sulfonate, which are not detected.
All percentages stated in the present invention are by mass unless otherwise indicated.
The compound of the formula III can be used as a reference substance for controlling the quality of carbazochrome sodium sulfonate and a preparation thereof and detecting the validity period of the carbazochrome sodium sulfonate and the preparation thereof. It is therefore another object of the present invention to provide the use of a compound of formula iii for the preparation of a control of carbazochrome sodium sulfonate drug substance or a formulation thereof.
One of the purposes of the invention is to provide a method for detecting the compound shown in the formula III, which uses high performance liquid chromatography and uses octadecylsilane chemically bonded silica as a filling agent; taking 0.12% ammonium dihydrogen phosphate solution-acetonitrile as a mobile phase; detection is carried out at wavelengths of 296nm, 280nm and 220nm, preferably at 220 nm.
Another object of the present invention is to provide a pharmaceutical preparation, which is composed of the pharmaceutical composition of the present invention and a pharmaceutically acceptable carrier.
The pharmaceutical formulations of the present invention may be in a variety of dosage forms well known in the art. The dosage form suitable for the present invention is selected from oral preparations, external preparations or injections, preferably oral preparations or injections, more preferably injections. The oral preparation is selected from oral solution, tablet, capsule, granule, pill, powder and syrup, preferably powder, tablet and granule; the tablet is a round or special-shaped tablet solid preparation which is prepared by uniformly mixing and pressing the medicament and proper auxiliary materials, and can be selected from common oral tablets, buccal tablets, sublingual tablets, buccal patches, chewable tablets, dispersible tablets, effervescent tablets, sustained release tablets, controlled release tablets or enteric-coated tablets and the like; the granules are selected from soluble granules (generally called granules), suspension granules, effervescent granules, enteric granules, sustained-release granules, controlled-release granules and the like; the external preparation is selected from gels, paste, liniments, lotions and smearing preparations, and preferably the paste is selected from emplastrum, cream or ointment; the injection is selected from injection, freeze-dried powder injection or sterile subpackaged preparation, and is preferably injection. The pharmaceutical formulations of the present invention may be prepared using formulation techniques well known in the art.
The pharmaceutically acceptable carrier according to the present invention is a commonly used excipient or adjuvant for preparing the formulation, which is well known in the art. The excipients or adjuvants commonly used in oral or external preparations include, but are not limited to, fillers (diluents), lubricants (glidants or antiadherents), dispersants, wetting agents, binders, solubilizers, antioxidants, and the like. Binders, such as syrup, cellulose and its derivatives, gelatin slurry, starch slurry, polyvinylpyrrolidone, PVP-K30, hydroxypropyl cellulose, methyl cellulose, pregelatinized starch, preferably povidone and cellulose derivatives, microcrystalline cellulose, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, ethyl cellulose or hydroxypropylmethyl cellulose; fillers, such as lactose, powdered sugar, dextrin, starch and derivatives thereof, cellulose and derivatives thereof, inorganic calcium salts, sorbitol, glycine, calcium sulphate dihydrate, microcrystalline cellulose, pregelatinized starch, mannitol, preferably lactose, dextrin; lubricants, such as aerosil, magnesium stearate, talc, aluminium hydroxide, boric acid, hydrogenated vegetable oils, polyethylene glycol, sodium lauryl sulphate, preferably magnesium stearate, silicon dioxide; disintegrants, for example starch or its derivatives, polyvinylpyrrolidone, microcrystalline cellulose, crospovidone, croscarmellose sodium, sodium carboxymethyl starch, low substituted hydroxypropyl cellulose, preferably starch derivatives such as sodium carboxymethyl starch, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch; wetting agents such as sodium lauryl sulfate, water or alcohols, and the like.
The excipients or adjuvants commonly used in the injection include, but are not limited to, antioxidants, for example, sulfur-containing compounds such as sulfite, bisulfite, sodium sulfite, sodium bisulfite, sodium metabisulfite, sodium thiosulfate; organic acids/alcohols/esters such as ascorbic acid, citric acid, malic acid, sorbitol, glycerol, propylene glycol, ascorbyl palmitate, esters such as hydroquinone, hydroxycoumarin, vitamin E; osmotic pressure regulators, such as sodium chloride, glucose, potassium chloride, magnesium chloride, calcium chloride, sorbitol, mannitol, and the like, preferably sodium chloride or sorbitol; a metal complexing agent, such as any one of or the combination of disodium edetate, tetrasodium edetate and calcium disodium edetate; a pH adjuster such as any one of hydrochloric acid, tartaric acid, sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, disodium hydrogen phosphate, acetic acid, sodium acetate, lactic acid, citric acid, sodium citrate, sodium bicarbonate, sodium carbonate, or a combination thereof; solubilizers such as tween-80, glycerol, propylene glycol, and the like; fillers or excipients, for example, lactose, mannitol, sorbitol, dextran, glycine and the like.
The injection is selected from injection, sterile powder for injection and concentrated solution for injection, and can be used for intramuscular injection, intravenous drip, etc. The specification of the injection of the invention is selected from lmL, 2mL, 5mL, 10mL, 20mL, 50mL, 100mL, 200mL, 250mL or 500 mL.
The pharmaceutical preparation of the present invention may be a unit preparation, and contains 10mg to 100mg of the pharmaceutical composition of the present invention as an essential active ingredient, preferably 10mg, 25mg, 30mg, 50mg, 100mg, in terms of carbazochrome sodium sulfonate trihydrate. The pharmaceutical composition of the present invention contains 5mg to 90mg, preferably 20mg, 40mg, 60mg, and 80mg, as an essential active ingredient, based on carbazochrome sodium sulfonate anhydride.
Another objective of the present invention is to provide a method for preparing injection, which is characterized in that the pharmaceutical composition of the present invention is mixed with a pharmaceutically acceptable injection carrier under anaerobic protection at a certain temperature and pH value range.
Further, the pharmaceutically acceptable injection carrier is selected from any one of an antioxidant, an osmotic pressure regulator, a solubilizer, a metal complexing agent and a pH regulator or the combination thereof.
Further, the antioxidant is, for example, a sulfur-containing compound such as sulfite, bisulfite, sodium sulfite, sodium bisulfite, sodium metabisulfite, sodium thiosulfate; organic acids/alcohols/esters such as ascorbic acid, citric acid, malic acid, sorbitol, glycerol, propylene glycol, ascorbyl palmitate, esters such as hydroxycoumarin, vitamin E; osmotic pressure regulators such as sodium chloride, glucose, potassium chloride, magnesium chloride, calcium chloride, sorbitol, mannitol, and the like; a metal complexing agent, such as any one of or the combination of disodium edetate, tetrasodium edetate and calcium disodium edetate; a pH adjuster such as any one of hydrochloric acid, tartaric acid, sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, disodium hydrogen phosphate, acetic acid, sodium acetate, lactic acid, citric acid, sodium citrate, sodium bicarbonate, sodium carbonate, or a combination thereof; solubilizers such as tween-80, glycerol, propylene glycol, and the like; fillers or excipients, for example, lactose, mannitol, sorbitol, dextran, glycine and the like.
The invention also aims to provide the application of the pharmaceutical composition or the pharmaceutical preparation thereof in preparing the hemostatic medicament.
Further, the hemostasis is selected from any one or a combination of bleeding tendency (e.g., purpura, etc.) caused by decrease of capillary resistance and increase of permeability, fundus hemorrhage exuded from skin or mucosa and intima due to decrease of capillary resistance, renal hemorrhage, and uterine hemorrhage.
Further, the hemostasis is selected from any one or combination of abnormal bleeding during and after surgery due to a decrease in capillary resistance.
Further, the hemostasis is selected from any one or combination of prevention and treatment of bleeding diseases of urinary system, upper gastrointestinal tract, respiratory tract and obstetrics and gynecology department and operation bleeding.
Compared with the prior art, the invention has the following advantages:
1) the invention separates and purifies the new compound shown in the formula III and carries out structure confirmation on the compound.
2) The invention separates and purifies the related substances possibly generated in the storage process of the carbazochrome sodium sulfonate to obtain the compound shown in the formula III, the adrenal color hydrazone and other related substances.
3) According to the invention, through intensive research, the carbazochrome sodium sulfonate pharmaceutical composition with high stability is obtained, namely, each link for preparing carbazochrome sodium sulfonate and a preparation thereof is controlled within a certain temperature and pH value range under the condition of oxygen-free protection, and the safety, effectiveness and stability of the carbazochrome sodium sulfonate raw material and the preparation are improved by effectively controlling the content of a compound in a formula III in the carbazochrome sodium sulfonate raw material and the preparation.
4) The invention also provides a preparation method of the compound of the formula III, and the prepared compound of the formula III is used as a reference substance for controlling the content of the compound of the formula III in the carbazochrome sodium sulfonate and the preparation thereof.
Drawings
Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:
FIG. 1 is an HPLC chart of a compound of formula III.
FIG. 2H of a compound of the formula III1NMR chart.
FIG. 3 is a mass spectrum of a compound having a structure represented by formula III.
Detailed Description
The following examples are further illustrative of the present invention and are in no way intended to limit the scope of the invention. The present invention is further illustrated in detail below with reference to examples, but it should be understood by those skilled in the art that the present invention is not limited to these examples and the preparation method used. Also, equivalent substitutions, combinations, improvements or modifications of the invention may be made by those skilled in the art based on the description of the invention, but these are included in the scope of the invention.
EXAMPLE 1 preparation of the Compound of formula III
Taking 10g of carbazochrome sodium sulfonate, adding 500ml of 50% ethanol water solution, heating, stirring and dissolving, adding 10ml of triethylamine, refluxing in water bath for 60 minutes, separating the reflux liquid by using a preparative high performance liquid phase, wherein the mobile phase is acetonitrile-ammonium dihydrogen phosphate buffer solution, and receiving the product flow to obtain 0.1g of the compound shown in the formula III. The content is not lower than 98 percent by HPLC peak area normalization method. MS (high resolution Mass Spectrometry) is 271.9961([ M + Na)]+)。
TABLE 1 HPLC MEASUREMENT RESULTS TABLE
| Retention time (minutes) | Peak area | Peak area (%) |
| 8.280 | 1109907760 | 98.61 |
| 12.533 | 15674886 | 1.39 |
| Total of | 1125582646 | 100.00 |
TABLE 21H NMR and1H-1h COSY determination analysis table
EXAMPLE 2 preparation of the Compound of formula III
Taking 10g of carbazochrome sodium sulfonate, adding 500ml of 50% ethanol water solution, heating, stirring and dissolving, adding 10ml of triethylamine, refluxing in a water bath for 60 minutes, adding 50g of chromatographic silica gel, drying under reduced pressure, then loading on a chromatographic column, eluting with methanol-ethyl acetate, receiving product flow, evaporating the solvent under reduced pressure to obtain a crude product, adding 100ml of 75% ethanol water solution, heating until the solid is completely dissolved, filtering while hot, cooling the filtrate, and precipitating crystals to obtain 0.1g of the compound shown in the formula III. The content is not lower than 98 percent by HPLC peak area normalization method. MS (LC-MS) is 226.2([ M-H)]-)。
TABLE 313CNMR, HSQC and HMBC assay data sheet
Example 3Toxicity Studies of Compounds of formula III
Using an L929 cell line (ATCC CCLl [ NCTC clone 929 (mouse fibroblast)) and carbazochrome sodium sulfonate (provided by jiangsu hanston pharmaceutical industry ltd., refined to 99.9% purity, not containing a compound of formula iii) and a compound of formula iii (provided by the present invention, purity 99.9%), incubating the cells with L929 for 24h, 48h, and 72h (final concentrations are 10, 5, 2.5, 1.25, 0.625, 0.3125, 0.15625, and 0.078125mg/ml), respectively, measuring the cell viability of each sample at different time points by a tetramethylazoazolium colorimetric method (MTT Assay), and calculating the half inhibitory concentration IC50 value at each time point by a Bliss method. As a result: the compound of formula III has an IC50 value of 3.87mg/ml and an IC50 value of 0.02 mg/ml. The result shows that the IC50 value of the carbazochrome sodium sulfonate is about 194 times that of the compound shown in the formula III, and the cytotoxicity of the compound shown in the formula III is far higher than that of the carbazochrome sodium sulfonate, so that the control of the content of the compound shown in the formula III in the carbazochrome sodium sulfonate bulk drug has important significance for improving the safety of the product.
Example 4Toxicity study of Carbazochrome sodium sulfonate and compositions thereof
By adopting an L929 cell strain, the weight ratio of the tested drug, namely carbazochrome sodium sulfonate (provided by Jiangsu Hanstongtong pharmaceutical industry Co., Ltd., refined into 99.9% purity and not containing a compound shown in a formula III) to the compound shown in the formula III (provided by the invention, purity is 99.9%) is respectively 1: 0.1%, 1: 0.25%, 1: 0.5%, 1: 0.75%, 1: 1%, 1: 1.5% and 1: 2% of the composition group (hereinafter referred to as "composition"), before administration, the test drug was dissolved in water for injection, and diluted to the desired concentration (wherein the final concentration of carbazochrome sodium sulfonate is 1mg/ml) according to the dosage requirement, and the cell viability of each test sample was measured at different time points by the tetramethylazoazolate colorimetry (MTT Assay).
TABLE 4 cytotoxicity response grading judgment Table
| Rank of | Relative survival Rate (%) |
| 0 | ≥100 |
| 1 | 80～99 |
| 2 | 50～79 |
| 3 | 30～49 |
| 4 | 0～29 |
TABLE 5 results of grading the cytotoxicity response of compositions containing carbazochrome sodium sulfonate and a compound of formula III
| Composition comprising a metal oxide and a metal oxide | Relative survival Rate (%) | Determining a level |
| 1：0.1％ | 83.53 | 1 |
| 1：0.25％ | 78.36 | 2 |
| 1：0.5％ | 69.14 | 2 |
| 1：0.75％ | 62.78 | 2 |
| 1：1％ | 56.22 | 2 |
| 1：1.5％ | 45.35 | 3 |
| 1:2％ | 33.41 | 3 |
As can be seen from Table 5, when the content of the compound of formula III is less than or equal to 1%, the cytotoxic reaction of the composition is not greater than 2 grade, which falls into the acceptable range; when the content of the compound III is 1.5%, the cytotoxicity reaction is grade 3, and the reaction belongs to positive reaction; when the content of the compound III is 2%, the relative survival rate of cells is only 33.41%, and although the cytotoxicity response is 3 grades, the cytotoxicity response is close to the value of 4 grades.
As a result, it is found that the content of the compound III is not more than 1%, and it is theoretically relatively safe. However, in order to further ensure the safety of administration, the content of the compound III in the composition should preferably not be higher than 0.5%.
Example 5
Preparation of Carbazochrome sodium sulfonate (hereinafter referred to as composition 1)
Under the protection of nitrogen, 500g of adrenal color hydrazone is put into a three-neck flask, then sodium bisulfite solution (sodium bisulfite 400g is dissolved in 1000ml of water) is added, the temperature is raised to 50 ℃, the mixture is stirred for 6 hours, filtrate is filtered and concentrated to 800ml, the mixture is frozen, crystals are separated out, the crystals are filtered, washed by 800ml of acetone, and 564g is obtained after drying at 50 ℃.
Under the protection of nitrogen, 560g of carbazochrome sodium sulfonate is added with 2000ml of 75% ethanol aqueous solution, the mixture is heated until the solid is completely dissolved, 12g of activated carbon is added, the mixture is stirred for 30 minutes, the mixture is filtered while the mixture is hot, the filtrate is cooled, crystals are separated out, the crystals are filtered, washed by 250ml of ethanol, and the crystals are dried at 50 ℃ to obtain 425.6g of carbazochrome sodium sulfonate. To obtain the carbazochrome sodium sulfonate composition 1.
EXAMPLE 6 preparation of pharmaceutical formulations
Prescription 2 of injection
Prescription 3 of injection
Prescription 4 of injection
Prescription 5 of injection
1. Preparation process
(1) Injecting 50% of injection water according to the prescription into a thick preparation tank, adding sodium chloride, sodium bisulfite and disodium edetate according to the prescription into the thick preparation tank successively under the protection of nitrogen gas at the temperature of not more than 40 ℃, stirring and dissolving, adding 0.05% of needle carbon according to the prescription, stirring for 30 minutes, filtering by using a titanium filter stick to remove carbon, and filtering into a thin preparation tank.
(2) Adding injection water with the amount of 80% of the prescription amount into a diluting preparation tank, continuously filling nitrogen for protection, adding the carbazochrome sodium sulfonate with the prescription amount at the temperature of not more than 40 ℃, stirring and dissolving, adjusting the pH value by using 2mol/L sodium hydroxide solution, adding the carbon for injection with the prescription amount of 0.02% (g/ml), adding the injection water to the full amount, stirring for 30 minutes, and filtering by using a titanium filter stick to remove the carbon.
(3) Detecting an intermediate product;
(4) fine filtering with 0.22 μm microporous membrane, bottling, plugging, and capping;
(5) sterilizing at 115 deg.C for 30 min, spraying hot water, rapidly cooling to 60 deg.C, taking out, and naturally cooling to room temperature.
(6) Light inspection;
(7) and packaging and warehousing, and fully inspecting finished products.
Tablet formulation 6
The preparation process comprises the following steps:
(1) adding carbazochrome sodium sulfonate, lactose, corn starch, microcrystalline cellulose and appropriate amount of water into a granulator, stirring, and granulating.
(2) Placing the prepared wet granules in an oven, drying at 60 deg.C, and controlling the water content below 5% at the final drying point.
(3) After the drying, the granules were sized with a 24 mesh screen.
(4) And (3) finishing granules, adding magnesium stearate and silicon dioxide according to the formula proportion, placing the mixture in a mixer, and measuring the content and the moisture of the granules after mixing.
(5) Tabletting, and packaging with aluminum-plastic bubble cap or glass bottle.
(6) And (5) putting the paper boxes into storage after being externally packaged, and fully inspecting finished products.
Example 7Detection of a compound of formula III in a pharmaceutical formulation
Measuring by high performance liquid chromatography, and using octadecylsilane chemically bonded silica as filler; 0.12 percent ammonium dihydrogen phosphate solution-acetonitrile (91:9) is taken as a mobile phase; the detection wavelength was 220 nm.
Precisely measuring a proper amount of sample (the injection is directly used, the tablet is a proper amount of sample, grinding, precisely weighing a proper amount of sample, and filtering the solution after ultrasonic dissolution by using a mobile phase), and quantitatively diluting the mobile phase to prepare a solution containing about 0.4mg of carbazochrome sodium sulfonate in each 1ml as a test solution; precisely measure 1ml, place in a 100ml measuring flask, dilute to the scale with mobile phase, shake well, as control solution. And dissolving an appropriate amount of adrenal color hydrazone reference substance and the compound of the formula III in a mobile phase, and quantitatively diluting to prepare solutions containing about 8 mu g of adrenal color hydrazone reference substance and 2 mu g of the compound of the formula III in each 1ml of solution respectively as reference substance solutions. Then, the test solution, the reference solution and the reference solution were measured precisely and 10. mu.l each, and the measured solution, the reference solution and the reference solution were injected into a liquid chromatograph, respectively, and the retention time of the chromatogram to the main component peak was recorded to 3 times. Wherein, the adrenal color hydrazone reference substance and the compound shown in the formula III are calculated by peak areas according to an external standard method.
Table 6 stability study of the formulations of the invention
As can be seen from Table 6, the pharmaceutical composition of the present invention can control the content of the related substances within a safe range, especially the content of the compound of formula III in the pharmaceutical composition within 0.5%, after 24 months of storage.
Claims (8)
1. An injection with good stability and high safety is characterized by being prepared from the following components: 80 parts of a pharmaceutical composition containing carbazochrome sodium sulfonate, 900 parts of sodium chloride, 1-100 parts of sodium bisulfite, 10 parts of edetate disodium, 100 parts by volume of water for injection, and adjusting the pH value to 5.5-6.0 by sodium hydroxide;
the medicinal composition containing carbazochrome sodium sulfonate comprises carbazochrome sodium sulfonate with the content of not less than 98%, a compound of a formula III with the content of not more than 0.5%, adrenal color hydrazone with the content of not more than 0.5%, and the balance of other related substances;
2. a tablet with good stability and high safety is characterized by being prepared from the following components: 30 parts of a pharmaceutical composition containing carbazochrome sodium sulfonate, 120 parts of lactose, 20 parts of corn starch, 28 parts of microcrystalline cellulose, 1 part of silicon dioxide and 1 part of magnesium stearate;
the medicinal composition containing carbazochrome sodium sulfonate comprises carbazochrome sodium sulfonate with the content of not less than 98%, a compound of a formula III with the content of not more than 0.5%, adrenal color hydrazone with the content of not more than 0.5%, and the balance of other related substances;
3. the injection according to claim 1 or the tablet according to claim 2, wherein the pharmaceutical composition containing carbazochrome sodium sulfonate is prepared by a method comprising: adding sodium bisulfite solution into adrenal color hydrazone under nitrogen protection, heating and stirring until reaction is complete, filtering, concentrating the filtrate, refrigerating, precipitating crystals, filtering, washing with acetone, and drying to obtain the final product.
4. A method for detecting a compound of formula III is characterized in that high performance liquid chromatography is adopted, and octadecylsilane chemically bonded silica is used as a filling agent; taking 0.12% ammonium dihydrogen phosphate solution-acetonitrile as a mobile phase; the detection is carried out by using the wavelengths of 296nm, 280nm and 220 nm.
5. The method of claim 4, wherein the detection is at a wavelength of 220 nm.
6. The application of the compound of the formula III in preparing a contrast product of a carbazochrome sodium sulfonate medicament or a preparation thereof.
7. Use of the injection of claim 1 or the tablet of claim 2 for the preparation of a hemostatic medicament.
8. A compound of formula III, having a chemical name: 1-methyl-6-hydroxyindole-2-sulfonic acid sodium salt, molecular formula: c9H8NNaO4S, the structural formula is as follows:
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| CN109884224B (en) * | 2019-03-06 | 2020-06-16 | 四川联成迅康医药股份有限公司 | Carbazochrome sodium sulfonate and method for measuring content of semicarbazide hydrochloride in preparation thereof |
| CN110478313B (en) * | 2019-05-17 | 2020-06-16 | 四川联成迅康医药股份有限公司 | Sodium carbazochrome injection |
| CN113476399B (en) * | 2021-07-30 | 2022-05-03 | 康普药业股份有限公司 | Sodium carbazochrome injection and preparation method thereof |
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