CN116350578A - Stable isoniazid injection and preparation method thereof - Google Patents

Abstract

The invention relates to the technical field of pharmaceutical preparations and discloses isoniazid injection and a preparation method thereof. The invention solves the problem that the content of free hydrazine in genotoxic impurities in isoniazid injection prepared by the prior art exceeds the ICH safety limit. The isoniazid injection provided by the invention is prepared by taking isoniazid as an active ingredient, taking anhydrous citric acid and citric acid hydrate composition, anhydrous citrate and citrate hydrate composition and water for injection as auxiliary materials, and performing the procedures of liquid preparation, filtration, filling and sealing, sterilization and packaging. The invention discovers that isoniazid injection with extremely low free hydrazine content can be obtained by adjusting the prescription composition and the dosage of auxiliary materials and further optimizing the preparation process, and the stability and the safety are better. After 2 years of storage at normal temperature, the free hydrazine content is still lower than the safety limit value of 200ppm.

Description

Stable isoniazid injection and preparation method thereof

Technical Field

The invention relates to the technical field of pharmaceutical preparations, in particular to a stable isoniazid injection and a preparation method thereof.

Background

Isoniazid is a compound which is synthesized for the first time in 1898 and has the chemical structure as follows:

。

scientists in 1952 have found that isoniazid has a significant effect on tubercle bacillus. Isoniazid injection is approved in japan on the market in the same year. Isoniazid injection is also approved in China, the United states, the United kingdom and other countries. Isoniazid is still the first line drug in current tuberculosis treatment. As an alternative to oral treatment, isoniazid injection is of clinical significance for the treatment of patients who cannot be orally administered.

The isoniazid injection prepared by the prior art has the condition that the content of free hydrazine of genotoxic impurities exceeds the safety limit value of ICH, and cannot guarantee safe medication. Genotoxic impurities are a class of impurities that react with DNA, cause DNA damage, induce gene mutations at very low levels, and may produce carcinogens. The effective control of genotoxic impurities is of great significance for drug safety. New researches are carried out on isoniazid injection by the manufacturers at home. Wherein, the patent CN111265475A provides a preparation method of isoniazid injection and isoniazid injection. Hydrochloric acid is added to the prescription of the product mentioned in the patent as a pH regulator, and sterilization is carried out by adopting a residual probability method of sterilizing at 121 ℃ for 8 minutes in the preparation process. The invention reduces free hydrazine which is a genotoxic impurity to a certain extent, but still fails to effectively solve the problems that the content of the free hydrazine is continuously increased and exceeds a safety limit value of 200ppm during the stability period, and the invention adopts a residual probability method of sterilizing at 121 ℃ for 8 minutes to ensure that the product has a certain potential risk in sterility. From the aspect of safety, the sterility assurance level is directly related to the safety of clinical use of the medicine, and the preparation process should be tried to be optimized as much as possible, so that the product is resistant to the excessive sterilization condition of high-pressure steam sterilization at 121 ℃ for 15-30 minutes, and the product has the highest sterility assurance level.

Disclosure of Invention

The invention aims at solving the problem that the content of free hydrazine in the genotoxic impurity in isoniazid injection is too high.

In order to solve the problem that the prior art means cannot control the content of free hydrazine which is a genotoxic impurity in isoniazid injection to exceed the ICH safety limit, the invention discloses a pharmaceutical composition with extremely low content of free hydrazine impurity and stable quality.

The invention has been made in order to solve the deficiency of the prior art, and as a result, it has been unexpectedly found that the above technical problems can be solved by the following technical scheme, thereby completing the invention.

The specific contents are as follows: an isoniazid injection comprises isoniazid, a stabilizer and water for injection, wherein the stabilizer is a composition of citric acid and citric acid salt, wherein the citric acid is any one or two of anhydrous citric acid and citric acid hydrate, and the citric acid salt is any one or two of anhydrous citric acid salt and citric acid salt hydrate.

Further, isoniazid in isoniazid injection is dissolved in water for injection and exists in a solution form, and the mass volume fraction of isoniazid is 5%.

Further, the molar concentration of the citric acid in the isoniazid injection is 0.007mol/L to 0.0275mol/L.

Further, the molar concentration of citrate in the isoniazid injection is 0.0725mol/L to 0.093mol/L.

Further, the anhydrous citrate may be selected from anhydrous sodium citrate and anhydrous potassium citrate, and the citrate hydrate may be selected from sodium citrate hydrate and potassium citrate hydrate.

Furthermore, the isoniazid injection must contain 0.007 mol/L-0.0275 mol/L anhydrous citric acid and citric acid hydrate and 0.0725 mol/L-0.093 mol/L citrate and citric acid hydrate.

Further, the isoniazid injection composition comprises water for injection added to 100% by volume, and the pH is 5.6-6.6.

In order to control the free hydrazine content in the isoniazid injection to be at an extremely low level, the inventor performs a great amount of analysis and exploration work, and finally discovers that after citric acid (anhydrous citric acid or citric acid hydrate) and citric acid salt (anhydrous sodium citrate or anhydrous potassium citrate, sodium citrate hydrate or potassium citrate hydrate) are added in a prescription to serve as pH buffering agents, the free hydrazine content of the isoniazid injection can be kept at 0 day and accelerated test stability for 6 months and long-term storage at 30 ℃ for no more than 200ppm, and the product quality stability and the medication safety of the isoniazid injection are greatly improved.

Isoniazid injection compositions include, but are not limited to, the following formulation process.

(1) Preparing liquid: dissolving 0.007-0.0275 mol of citric acid (anhydrous citric acid and citric acid hydrate) and 0.0725-0.093 mol of citric acid salt (anhydrous sodium citrate, anhydrous potassium citrate, sodium citrate hydrate and potassium citrate hydrate) in 600 milliliters of water for injection cooled to 30-40 ℃. Introducing high-purity nitrogen into the solution, maintaining the dissolved oxygen at 1 mg/L-2 mg/L, then adding 50g of isoniazid into the solution, stirring and dissolving completely, and adding the isoniazid into the total volume to 1 liter by injection cooled to 30-40 ℃.

(2) And (3) filtering: the liquid medicine is filtered by the dosing tank through the 0.45 micron filter element to the intermediate tank, then is filtered by the intermediate tank through the 0.22 micron filter element to the sterile tank, and the liquid medicine in the sterile tank is filtered by the 0.22 micron filter element and then is filled. The first filter element with the diameter of 0.45 micrometers can reduce the microorganism load in the liquid medicine, and provides sufficient guarantee for the subsequent filter element with the diameter of 0.22 micrometers to completely intercept microorganisms in the liquid medicine.

(3) And (3) filling and sealing: 2.3-2.5 ml of liquid medicine is filled in a 2ml borosilicate glass ampoule, nitrogen is filled before and after the ampoule is filled, and the nitrogen filling time is 1-5 seconds. Nitrogen is filled before and after filling, so that isoniazid can be well prevented from being oxidized, and the color of the product is near colorless.

(4) And (3) sterilization: and sterilizing the intermediate product after the melt sealing by a water bath type sterilizing cabinet. Sterilizing at 121 ℃ for 15-30 minutes. And controlling the cooling time to be 8 minutes after sterilization.

(5) After sterilization, 100% leak detection and lamp inspection are carried out, thus obtaining the injection.

The innovation and positive significance of the invention are as follows: 1) The invention provides a formula of isoniazid injection with high stability and safety and a preparation method thereof, wherein citric acid (anhydrous citric acid and citric acid hydrate) and citric acid salt (anhydrous citric acid and citric acid hydrate) used in the formula can effectively increase the stability of products, and the safety level of free hydrazine content of genotoxic impurities is controlled below 200ppm within the 2-year effective period from production to room temperature storage. 2) The invention provides a new preparation method of isoniazid injection. Through nitrogen charging protection in the liquid preparation and encapsulation processes and control of the cooling time in the sterilization process, the quality of the product is still stable under the condition of excessive sterilization under the condition of resisting sterilization at 121 ℃ for 15-30 minutes. Compared with a residual probability method of sterilizing at 121 ℃ for 8 minutes, the method greatly improves the sterility assurance level of the product and further improves the clinical use safety of the product.

Detailed Description

Example 1

Table 1: isoniazid injection prescription composition and dosage form

Composition of components	Dosage of
		Isoniazid	50g
Citric acid monohydrate	0.0275mol
		Sodium citrate dihydrate	0.0725mol
Water for injection	To 1L

Raw materials and auxiliary materials were weighed according to the prescription composition and the amounts shown in table 1. The weighed citric acid monohydrate and sodium citrate dihydrate were dissolved in 600ml of water for injection cooled to 40 ℃. High-purity nitrogen is introduced into the liquid medicine until the dissolved oxygen is 1.0mg/L. Adding the weighed isoniazid, and stirring until the isoniazid is completely dissolved. The volume was set to 1000ml with cooled water for injection. The liquid medicine is protected by nitrogen after the volume is fixed. The liquid medicine is filtered by a filter element made of polyethersulfone with the diameter of 0.45 micrometers and two filter elements made of polyethersulfone with the diameter of 0.22 micrometers after the volume is fixed. Filling 2.40ml of the mixture into 2ml of medium borosilicate glass ampoule, and filling nitrogen for 2 seconds before and after filling the glass ampoule. The product is subjected to wet heat sterilization at 121 ℃ for 15 minutes after the sealing. Cooling time is 8 minutes after sterilization.

Example 2

Table 2: isoniazid injection prescription composition and dosage form

Composition of components	Dosage of
		Isoniazid	50g
Citric acid monohydrate	0.019mol
		Sodium citrate dihydrate	0.081mol
Water for injection	To 1L

Raw materials and auxiliary materials were weighed according to the prescription composition and the amounts shown in table 2. The weighed citric acid monohydrate and sodium citrate dihydrate were dissolved in 600ml of water for injection cooled to 40 ℃. High-purity nitrogen is introduced into the liquid medicine until the dissolved oxygen is 1.0mg/L. Adding the weighed isoniazid, and stirring until the isoniazid is completely dissolved. The volume was set to 1000ml with cooled water for injection. The liquid medicine is protected by nitrogen after the volume is fixed. The liquid medicine is filtered by a filter element made of polyethersulfone with the diameter of 0.45 micrometers and two filter elements made of polyethersulfone with the diameter of 0.22 micrometers after the volume is fixed. Filling 2.40ml of the mixture into 2ml of medium borosilicate glass ampoule, and filling nitrogen for 2 seconds before and after filling the glass ampoule. The product is subjected to wet heat sterilization at 121 ℃ for 15 minutes after the sealing. Cooling time is 8 minutes after sterilization.

Example 3

Table 3: isoniazid injection prescription composition and dosage form

Composition of components	Dosage of
		Isoniazid	50g
Citric acid monohydrate	0.007mol
		Anhydrous sodium citrate	0.093mol
Water for injection	To 1L

Raw materials and auxiliary materials were weighed according to the prescription composition and the amounts shown in table 3. The weighed citric acid monohydrate and anhydrous sodium citrate were dissolved in 600ml of water for injection cooled to 40 ℃. High-purity nitrogen is introduced into the liquid medicine until the dissolved oxygen is 1.0mg/L. Adding the weighed isoniazid, and stirring until the isoniazid is completely dissolved. The volume was set to 1000ml with cooled water for injection. The liquid medicine is protected by nitrogen after the volume is fixed. The liquid medicine is filtered by a filter element made of polyethersulfone with the diameter of 0.45 micrometers and two filter elements made of polyethersulfone with the diameter of 0.22 micrometers after the volume is fixed. Filling 2.40ml of the mixture into 2ml of medium borosilicate glass ampoule, and filling nitrogen for 2 seconds before and after filling the glass ampoule. The product is subjected to wet heat sterilization at 121 ℃ for 15 minutes after the sealing. Cooling time is 8 minutes after sterilization.

Example 4

Table 4: isoniazid injection prescription composition and dosage form

Composition of components	Dosage of
		Isoniazid	50g
Citric acid anhydrous	0.019mol
		Sodium citrate dihydrate	0.081mol
Water for injection	To 1L

Raw materials and auxiliary materials were weighed according to the prescription composition and the amounts shown in table 4. The weighed anhydrous citric acid and sodium citrate dihydrate were dissolved in 600ml of water for injection cooled to 30 ℃. High-purity nitrogen is introduced into the liquid medicine until the dissolved oxygen is 1.0mg/L. Adding the weighed isoniazid, and stirring until the isoniazid is completely dissolved. The volume was set to 1000ml with cooled water for injection. The liquid medicine is protected by nitrogen after the volume is fixed. The liquid medicine is filtered by a filter element made of polyethersulfone with the diameter of 0.45 micrometers and two filter elements made of polyethersulfone with the diameter of 0.22 micrometers after the volume is fixed. Filling 2.40ml of the mixture into 2ml of medium borosilicate glass ampoule, and filling nitrogen for 2 seconds before and after filling the glass ampoule. The product is subjected to wet heat sterilization at 121 ℃ for 15 minutes after the sealing. Cooling time is 8 minutes after sterilization.

Example 5

Table 5: isoniazid injection prescription composition and dosage form

Composition of components	Dosage of
		Isoniazid	50g
Citric acid monohydrate	0.019mol
		Sodium citrate dihydrate	0.081mol
Water for injection	To 1L

Raw materials and auxiliary materials were weighed according to the prescription composition and the amounts shown in table 5. The weighed citric acid monohydrate and sodium citrate dihydrate were dissolved in 600ml of water for injection cooled to 30 ℃. High-purity nitrogen is introduced into the liquid medicine until the dissolved oxygen is 2.0mg/L. Adding the weighed isoniazid, and stirring until the isoniazid is completely dissolved. The volume was set to 1000ml with cooled water for injection. The liquid medicine is protected by nitrogen after the volume is fixed. The liquid medicine is filtered by a filter element made of polyethersulfone with the diameter of 0.45 micrometers and two filter elements made of polyethersulfone with the diameter of 0.22 micrometers after the volume is fixed. Filling 2.40ml of the mixture into 2ml of medium borosilicate glass ampoule, and filling nitrogen for 2 seconds before and after filling the glass ampoule. The product is subjected to wet heat sterilization at 121 ℃ for 15 minutes after the sealing. Cooling time is 8 minutes after sterilization.

Example 6

This example differs from example 2 in that sodium citrate dihydrate is changed to potassium citrate monohydrate, the remainder being identical.

Comparative example 1

Table 6: isoniazid injection prescription composition and dosage form

Composition of components	Dosage of
		Isoniazid	50g
Sodium chloride	9g
		Water for injection	To 1L

Raw materials and auxiliary materials were weighed according to the prescription composition and the amounts shown in table 4. The weighed sodium chloride was dissolved in 600ml of water for injection cooled to 40 ℃. High-purity nitrogen is introduced into the liquid medicine until the dissolved oxygen is 1.0mg/L. Adding the weighed isoniazid, and stirring until the isoniazid is completely dissolved. The volume was set to 1000ml with cooled water for injection. The liquid medicine is protected by nitrogen after the volume is fixed. The liquid medicine is filtered by a filter element made of polyethersulfone with the diameter of 0.45 micrometers and two filter elements made of polyethersulfone with the diameter of 0.22 micrometers after the volume is fixed. Filling 2.40ml of the mixture into 2ml of medium borosilicate glass ampoule, and filling nitrogen for 2 seconds before and after filling the glass ampoule. The product is subjected to wet heat sterilization at 121 ℃ for 15 minutes after the sealing. Cooling time is 60 minutes after sterilization.

Comparative example 2

This comparative example differs from comparative example 1 in that the post-sterilization cool down time was shortened from 60 minutes to 8 minutes.

Comparative example 3

Table 7: isoniazid injection prescription composition and dosage form

Composition of components	Dosage of
		Isoniazid	50g
Sodium chloride	9g
		Hydrochloric acid	Proper amount of
Water for injection	To 1L

Raw materials and auxiliary materials were weighed according to the prescription composition and the amounts shown in table 5. The weighed sodium chloride was dissolved in 600ml of water for injection cooled to 40 ℃. High-purity nitrogen is introduced into the liquid medicine until the dissolved oxygen is 1.0mg/L. Adding the weighed isoniazid, and stirring until the isoniazid is completely dissolved. The pH was adjusted to 6.0 with hydrochloric acid and the volume was set to 1000ml with chilled water for injection. The liquid medicine is protected by nitrogen after the volume is fixed. The liquid medicine is filtered by a filter element made of polyethersulfone with the diameter of 0.45 micrometers and two filter elements made of polyethersulfone with the diameter of 0.22 micrometers after the volume is fixed. Filling 2.40ml of the mixture into 2ml of medium borosilicate glass ampoule, and filling nitrogen for 2 seconds before and after filling the glass ampoule. The product is subjected to wet heat sterilization at 121 ℃ for 15 minutes after the sealing. Cooling time is 8 minutes after sterilization.

Comparative example 4

Table 8: isoniazid injection prescription composition and dosage form

Composition of components	Dosage of
		Isoniazid	50g
Citric acid monohydrate	0.041mol
		Sodium citrate dihydrate	0.059mol
Water for injection	To 1L

Raw materials and auxiliary materials were weighed according to the prescription composition and the amounts shown in table 6. The weighed citric acid monohydrate and sodium citrate dihydrate were dissolved in 600ml of water for injection cooled to 40 ℃. High-purity nitrogen is introduced into the liquid medicine until the dissolved oxygen is 1.0mg/L. Adding the weighed isoniazid, and stirring until the isoniazid is completely dissolved. The volume was set to 1000ml with cooled water for injection. The liquid medicine is protected by nitrogen after the volume is fixed. The liquid medicine is filtered by a filter element made of polyethersulfone with the diameter of 0.45 micrometers and two filter elements made of polyethersulfone with the diameter of 0.22 micrometers after the volume is fixed. Filling 2.40ml of the mixture into 2ml of medium borosilicate glass ampoule, and filling nitrogen for 2 seconds before and after filling the glass ampoule. The product is subjected to wet heat sterilization at 121 ℃ for 15 minutes after the sealing. Cooling time is 8 minutes after sterilization.

The data of examples and comparative examples were subjected to comparative analysis using the properties, pH, related substances, and free hydrazine as main evaluation indexes:

table 9: isoniazid injection product quality contrast (0 days)

	Traits (3)	pH value of	Total impurity of related substances (%)	Free hydrazine (ppm)
					Example 1	Colorless clear liquid	5.6	0.21	100
Example 2	Colorless clear liquid	6.0	0.19	91
					Example 3	Colorless and colorlessClear liquid	6.6	0.21	94
Example 4	Colorless clear liquid	6.0	0.18	97
					Example 5	Colorless clear liquid	6.0	0.19	92
Example 6	Colorless clear liquid	6.0	0.19	95
					Comparative example 1	Yellowish clear liquid	7.0	0.35	350
Comparative example 2	Colorless clear liquid	7.0	0.25	160
					Comparative example 3	Colorless clear liquid	6.0	0.20	88
Comparative example 4	Colorless clear liquid	5.1	0.23	97

Table 10: isoniazid injection product quality contrast (6 months acceleration)

	Traits (3)	pH value of	Total impurity of related substances (%)	Free hydrazine (ppm)
					Example 1	Colorless clear liquid	5.6	0.37	187
Example 2	Colorless clear liquid	6.0	0.33	156
					Example 3	Colorless clear liquid	6.6	0.36	179
Example 4	Colorless clear liquid	6.0	0.33	149
					Example 5	Colorless clear liquid	6.0	0.32	160
Example 6	Colorless clear liquid	6.0	0.32	158
					Comparative example 1	Yellowish clear liquid	6.8	1.8	5732
Comparative example 2	Yellowish clear liquid	6.9	1.7	5404
					Comparative example 3	Yellowish clear liquid	6.1	0.85	2031
Comparative example 4	Colorless clear liquid	5.1	0.51	289

Remarks: the acceleration conditions were: 40 ℃/75% RH

Table 11: isoniazid injection product quality contrast (24 months long term)

	Traits (3)	pH value of	Total impurity of related substances (%)	Free hydrazine (ppm)
					Example 1	Colorless clear liquid	5.6	0.21	109
Example 2	Colorless clear liquid	6.0	0.19	95
					Example 3	Colorless clear liquid	6.6	0.21	106
Example 4	Colorless clear liquid	6.0	0.18	104
					Example 5	Colorless clear liquid	6.0	0.19	112
Example 6	Colorless clear liquid	6.0	0.20	102
					Comparative example 1	Colorless clear liquid	6.8	1.0	2200
Comparative example 2	Colorless clear liquid	6.9	0.90	2054
					Comparative example 3	Colorless clear liquid	6.1	0.70	1325
Comparative example 4	Colorless clear liquid	5.1	0.49	248

Remarks: the long-term conditions are as follows: 30 ℃/65% RH

The data comparison result shows that the isoniazid injection provided by the invention can effectively control the free hydrazine content of genotoxic impurities in the sterilized product, and the free hydrazine content can still be controlled below 200ppm in the stability investigation period. The invention has important significance for the stability of the product quality and the safety in clinical use.

The examples set forth herein are merely illustrative of the present invention to solve the deficiencies of the prior art. It will be apparent to those skilled in the art that several modifications may be made without departing from the principles of the invention, and such modifications are intended to be within the scope of the invention.

Claims (7)

1. The isoniazid injection is characterized by comprising isoniazid, a stabilizer and water for injection, wherein the stabilizer is a composition of citric acid and citric acid salt, the citric acid is any one or two of anhydrous citric acid and citric acid hydrate, and the citric acid salt is any one or two of anhydrous citric acid salt and citric acid salt hydrate.

2. Isoniazid injection according to claim 1 characterized in that isoniazid mass fraction is 5%.

3. Isoniazid injection according to claim 1 characterized in that the molar concentration of citric acid is 0.007 mol/L-0.0275 mol/L.

4. Isoniazid injection according to claim 1 characterized in that the molar concentration of citrate is 0.0725mol/L to 0.093mol/L.

5. The isoniazid injection according to claim 1 wherein said anhydrous citrate is selected from the group consisting of sodium citrate anhydrous and potassium citrate anhydrous.

6. Isoniazid injection according to claim 1, characterized in that said citrate hydrate is selected from sodium citrate hydrate and potassium citrate hydrate.

7. Isoniazid injection according to claims 1-6, characterized in that its preparation method comprises the following steps:

(1) Dissolving a stabilizer in water for injection cooled to 30-40 ℃;

introducing high-purity nitrogen into the solution to maintain 1-2 mg/L of dissolved oxygen in the solution, then adding isoniazid into the solution, stirring and dissolving completely, and fixing the volume by using water for injection cooled to 30-40 ℃;

(2) Filtering the liquid medicine from a batching tank to a middle tank through a 0.45-micrometer filter element, filtering the liquid medicine from the middle tank to an aseptic tank through a 0.22-micrometer filter element, filtering the liquid medicine from the aseptic tank through a 0.22-micrometer filter element, and filling;

(3) Filling 2.3-2.5 ml of liquid medicine into a 2ml borosilicate glass ampoule, and filling nitrogen into the ampoule before and after filling;

(4) Sterilizing the intermediate product after the melt sealing by a water bath type sterilizing cabinet, sterilizing for 15-30 minutes at 121 ℃, and cooling for 8 minutes after sterilizing;

(5) After sterilization, 100% leak detection and lamp inspection are carried out, thus obtaining the injection.