CN102697739B - Preparation method of powder injection for reducing tigecycline epimer - Google Patents

Abstract

The invention discloses a preparation method of a powder injection for reducing tigecycline epimer. The preparation method comprises the following steps of: taking the tigecycline and fined lactose, adding water for injection to dissolve the materials, firstly dissolving the materials by using acid, enabling the pH of the solution to be proper, then readjusting the pH to be proper by using alkaline, filtering with a 0.22mum microfiltration membrane, carrying out subpackage, semi-plug-pressing, freeze-drying, plug pressing and rolled covering to obtain the powder injection. The pH adjusting agent is one or more of inorganic acids; and the pH readjusting agent is one or more of inorganic alkalines. The tigecycline powder injection prepared by the preparation method disclosed by the invention has the advantages that the content of impurities is low, the mass is stable in the putting process and the clinical use is safer and more effective.

Description

A kind of injectable powder preparation method that reduces tigecycline epimer

Technical field

The invention belongs to pharmaceutical field, relate to a kind of preparation method of tigecycline injectable powder, relate in particular to a kind of injectable powder preparation method that can reduce tigecycline epimer.

Background technology

Tigecycline is first approved intravenous injection Tigecycline class antibiotic, contains a glycyl amino, is replaced in 9 of minocycline.This replacement form does not see any natural or semi-synthetic tetracycline compound, thereby gives the Microbiological Characteristics of tigecycline uniqueness.Tigecycline is not subject to the impact of the large resistance mechanism of Tetracyclines two (ribosome protection and outer row's mechanism).Correspondingly, in vitro and in vivo test confirms that tigecycline has broad spectrum antibiotic activity.Not yet find that tigecycline and other antibiotic exist crossing drug resistant.Tigecycline is not subject to beta lactamase (comprising extended spectrum β lactamases), target position modification, Macrolide efflux pump or enzyme target position to change the impact of resistance mechanisms such as (as gyrase/topoisomerases).In vitro study does not confirm that tigecycline and other Common Antibiotics exist antagonism.Its mechanism of action is to be combined with 30S ribosome A position, stops amino acid transport RNA to enter ribosome, thereby has stoped amino acid residue to form peptide chain, and then kills or anti-bacteria breeding.

Tigecycline is mainly used in that the complexity intracavity that treatment can Portugal bacterium causes by Grain-negative or positive pathogen, anaerobe and Methicillin-resistant Staphylococcus aureus and methicillin-sensitivity infects, in lung coccus infect, complicated skin and soft tissue infection thereof etc.Compare the advantage such as tigecycline has has a broad antifungal spectrum, be difficult for to produce drug resistance, long half time, consumption are little with other antibiotic.Tigecycline Xiang doctors provide a kind of new, can be at treatment initial stage selective broad ectrum antibiotic when the cause of disease not yet understands, and do not need to adjust dosage according to impaired renal function situation, easy to use, use in every 12 hours once.

Tigecycline is by former the grinding of Hui Shi (Wyeth), and in granted listing in 2005, trade name Tygacil, dosage form is lyophilized injectable powder.This drug stabilisation is poor, and its solution is oxidative degradation and generation epimer very easily, therefore, should do one's utmost to avoid the generation of above-mentioned impurity when preparing freeze-dried powder.

Research shows, when tigecycline is in the water of pH approximately 7.8 during lyophilizing, the lyophilizing block major impurity obtaining is oxidation Decomposition product, and in the water of lower pH value during lyophilizing, epimerism is turned to main degradation pathway and occurs, meanwhile, tigecycline has different epimerization characteristics from other tetracycline antibiotics, and under the condition of lower pH value, the epimerization of tigecycline is more remarkable.Therefore, in the time of at present mainly by lyophilizing tigecycline the lower pH value in the situation that, control epimerization product, or the method for controlling oxidative breakdown product in the situation that pH is 7.7-8.2 when lyophilizing tigecycline increases the stability of tigecycline.

The Chinese invention patent application 200680006447.3 of Wyeth provide a kind of under acid condition lyophilizing tigecycline, add carbohydrate simultaneously and increase the method for the stability of tigecycline as lactose, glucose etc.Under the condition in pH value acidity, oxidative degradation has been reduced to minimum degree, therefore adds the generation that carbohydrate is mainly used in avoiding epimer.Yet use tigecycline lyophilized formulations prepared by the method carry out 6 months 40 ℃ keep sample after accelerated test, its epimerism body burden has increased approximately 1.5%, therefore, in producing as tigecycline, do not wish to exist or exist the more few better impurity of content, the epimerism body burden of control tigecycline that should be stricter, to increase the stability of tigecycline lyophilized formulations.

The Chinese invention patent application 200680021384.9 of Wyeth disclose a kind of in the situation that pH is 7.7-8.2 the method for lyophilizing tigecycline, in the situation that epimerization reaction is reduced to minimum degree, by spraying in the preparation the noble gases such as a large amount of nitrogen to control oxygen content lower than 5ppm and to keep the temperature of water to reduce oxidative breakdown product in the method for about 2-8 degree, reach the object of the stable tigecycline lyophilized formulations of preparation.Chinese invention patent application 200610098366.2 also points out, thereby replaces oxygen in the water for injection that dissolves tigecycline and reach and in preparation process, protect tigecycline not oxidized by being filled with the noble gases such as a large amount of nitrogen or argon.Yet these two kinds of method complex process are with high costs.

Chinese invention patent 200610096514.7 provides a kind of antioxidant that adds to solve tigecycline oxidative degradation problem, owing to adding oxidant generally to have Hidden Trouble In Clinical Medication Security problem in injection, and is unworthy advocating.

Summary of the invention:

The object of the invention is to, provide a kind of by regulating the method for pH value to reduce the lyophilized injectable powder preparation method of tigecycline epimerism body burden; Another object of the present invention is, the tigecycline injectable powder preparation method providing is when controlling epimer and increasing, and the basic zero growth of all the other known impurities and unknown impuritie, has greatly reduced and increased the hidden danger of bringing to patient's drug safety because of impurity.

The preparation method of tigecycline freeze-dried powder provided by the invention, can make tigecycline epimer growth rate obviously reduce, in 40 ℃ keep sample acceleration experiment in six months, epimer increment can be controlled at below 1.0%, under optimal conditions, even can be controlled in 0.3%.

The present invention is achieved through the following technical solutions:

A kind of injectable powder preparation method that reduces tigecycline epimer, comprise the following steps: get tigecycline, refined lactose, be dissolved in water for injection, first with acid, adjust and make material dissolution and make the pH of solution to suitable, then with alkali, return and adjust pH to suitable, 0.22um filtering with microporous membrane, subpackage, half tamponade, lyophilizing, tamponade, Zha Gai, obtains.

Reduce an injectable powder preparation method for tigecycline epimer, described acid be medicinal hydrochloric acid, sulphuric acid, phosphoric acid, etc. mineral acid.Described alkali is the inorganic bases such as medicinal sodium hydroxide, potassium hydroxide.

Reduce an injectable powder preparation method for tigecycline epimer, it is 1.0 to 4.4 that pH scope is adjusted in described acid, and it is 1.0-1.5 that preferred acid is adjusted pH scope

Reduce an injectable powder preparation method for tigecycline epimer, described alkali returns and adjusts pH scope is 4.5 to 6.3, and preferred bases is returned and adjusted pH scope is 5.2-6.3

Owing to preparing tigecycline lyophilized formulations under the present invention's condition that is 4.5-6.3 at pH value, therefore epimerism turns to the main degradation pathway of tigecycline, and can make the content of 40 ℃ of trimestral epimers that keep sample of this product increase from 1.0% by above-mentioned alkali callbacks, is minimumly down to 0.11%; The content of 40 ℃ of epimers that keep sample six months increases from 1.0% is minimumly down to 0.25%.

The embodiment of the present invention 1 is with reference to the preparation method of tigecycline injectable powder technique in Wyeth patent (ZL 200680006447.3), prepare respectively pH and be 5.60,5.20,4.80 tigecycline solution, 40 ℃ of epimer increments that keep sample six months of this product are all in 1.5% left and right.Compare with lyophilized formulations prepared by alkali callbacks of the present invention, the lyophilized formulations preparing keeps sample experiment in the time of 0 day at 40 ℃, both differences are not remarkable, and along with the time of 40 ℃ of experiments that keep sample lengthens, the recruitment of both epimers has the difference of significance.

By product research of the present invention is found, 40 ℃ of increases of accelerating the epimer under experiment of this technique mainly occur in the first two months, the quality keeping sample for a long time every 6 months is slightly better than 40 ℃ of characteristics of month simultaneously, under optimum condition, the epimer recruitment of this product acceleration after 6 months is lower than 0.3%, therefore can predict, the recruitment of this product epimer of long-term 24 months can be lower than the recruitment of accelerating 4 months, also can be lower than 0.3%.

Therefore, can find out, the present invention is better than Wyeth patent (ZL 200680006447.3) part and is, the present invention can slow down the speed that epimerization changes significantly, has improved largely the stability in the long-term storage process of tigecycline.

Research shows, the present invention by carrying out alkali callbacks under the environment of acid ph value, the increase of product stability is not because introduced the inorganic salts such as sodium chloride, sodium sulfate, potassium sulfate when regulating pH value, and the simple inorganic salt that adds cannot make its stability improve.The salt (as sodium chloride, potassium sulfate, sodium phosphate) that may bring into while regulating pH value by adding, investigate salt as shown in table 1 below on the concrete outcome of the impact of the epimerism body burden of the tigecycline of lyophilizing under pH5.2 condition:

Group is 1 for pH value is to pull back to the content of the sodium chloride producing at 4.0 o'clock at 4.8 o'clock by alkali, group 4 and organize 5 for pH value be by alkali, to pull back to the potassium sulfate that produces and the content of sodium phosphate at 4.8 o'clock at 4.0 o'clock.

Group

Inventory

0 day

One month

Two months

Three months

Six months

1

0.02mg (sodium chloride)

0.253％

0.697％

1.037％

1.277％

1.819％

2

0.1mg (sodium chloride)

0.246％

0.687％

1.054％

1.282％

1.857％

3

0.5mg (sodium chloride)

0.275％

0.689％

1.033％

1.254％

1.825％

4

0.03mg (potassium sulfate)

0.261％

0.708％

1.041％

1.285％

1.834％

5

0.02mg (sodium phosphate)

0.258％

0.716％

1.068％

1.269％

1.849％

As can be seen from the above table, even than group 1 drop into more the sodium chloride of volume (as organize 2 and group 3), also cannot slow down advancing the speed of tigecycline epimer, therefore, in the situation that not using alkali callbacks, add separately salt (as sodium chloride, potassium sulfate and potassium phosphate) cannot improve significantly the stability of product.

When the alkali callbacks that the present invention uses is prepared tigecycline lyophilized formulations, when controlling epimerism body burden, the basic zero growth of all the other known impurities and unknown impuritie.From accompanying drawing 1-3, by by pH value 1.5 by alkali pull back to pH5.2 the tigecycline lyophilized formulations prepared of method, 0 day, 3 months, 6 months 40 ℃, accelerate not occur obvious impurity peaks on experiment HPLC collection of illustrative plates.

Research shows, by alkali callbacks, prepares tigecycline lyophilized formulations, and difference is returned the formulation and technology of adjusting starting point, and the growth of epimer is also had to obvious impact.Alkali adjusts starting point pH value between 1.0-1.5, and stability can reach optimum, and tigecycline epimer recruitment can be controlled in 0.3%, and alkali adjusts starting point pH value lower than 1.0, and freeze-dried powder molding, color and luster is not good enough.

The invention provides a kind of method of preparing tigecycline lyophilized formulations by alkali callbacks, pass through the method, can slow down significantly advancing the speed of tigecycline epimer, while all the other known impurities of tigecycline and the basic zero growth of unknown impuritie, improved largely the stability in tigecycline storage process, made patient's medication safer, effective; And without inflated with nitrogen, process in production process, reduced cost.

Accompanying drawing explanation:

Fig. 1 for by pH value 1.5 by alkali pull back to pH5.2 40 ℃ of HPLC collection of illustrative plates that keep sample 0 day of the tigecycline lyophilized formulations prepared of method.

Fig. 2 for by pH value 1.5 by alkali pull back to pH5.2 40 ℃ of trimestral HPLC collection of illustrative plates that keep sample of the tigecycline lyophilized formulations prepared of method.

Fig. 3 for by pH value 1.5 by alkali pull back to pH5.2 40 ℃ of HPLC collection of illustrative plates that keep sample six months of the tigecycline lyophilized formulations prepared of method.

Fig. 4 is that pH value is 40 ℃ of trimestral HPLC collection of illustrative plates that keep sample of 5.2 tigecycline lyophilized formulations

The specific embodiment:

Embodiment 1:

With reference to the preparation method of tigecycline injectable powder technique in Wyeth patent (ZL 200680006447.3), prepare respectively pH and be 5.60,5.20,4.80 tigecycline solution, as shown in the group 1 of table 2, group 2 and group 3.

Use alkali callbacks to prepare tigecycline injectable powder, concrete technology is as follows:

Get tigecycline 10.03g, refined lactose 20.00g, inject the about 700ml of water and dissolve, extremely suitable with the hydrochloric acid conditioning solution pH of 5mol/L, then with 2mol/L sodium hydroxide solution, return and adjust pH to suitable, add water for injection to be diluted to 800ml, shake up, use 0.22um filtering with microporous membrane, subpackage, half tamponade, lyophilizing, tamponade, Zha Gai, obtains this product.Specifically as shown in the group 4-group 13 of table 2.

Table 2: tigecycline injectable powder stability test:

Experimental group	PH value	0 day	40 ℃ one month	40 ℃ two months	40 ℃ three months	40 ℃ six months
							1	4.8	0.364	0.654	1.010	1.206	1.805
2	5.2	0.415	0.788	1.131	1.311	1.866
							3	5.6	0.388	0.823	1.120	1.373	1.926
4	4.0 pull back to 5.2	0.226	0.494	0.702	0.867	1.241
							5	3.5 pull back to 5.2	0.230	0.455	0.541	0.728	1.116
6	3.0 pull back to 5.2	0.223	0.433	0.515	0.593	0.977
							7	1.5 pull back to 5.2	0.205	0.329	0.336	0.369	0.479
8	1.0 pull back to 5.2	0.204	0.275	0.339	0.415	0.495
							9	1.2 pull back to 5.2	0.211	0.281	0.308	0.322	0.457
10	1.3 pull back to 5.2	0.220	0.288	0.315	0.335	0.465
							11	1.4 pull back to 5.2	0.200	0.311	0.328	0.361	0.472
12	3.0 pull back to 6.3	0.201	0.418	0.476	0.553	0.952
							13	1.2 pull back to 6.3	0.208	0.277	0.331	0.386	0.501

As seen from the above table: the tigecycline injectable powder of preparing by pH alkali callbacks, its epimer recruitment can basic controlling below 1.0%, when pH value readjustment starting point is 1.0-1.5, tigecycline epimer recruitment can be controlled at below 0.3%.

Embodiment 2:

Get tigecycline 10.03g, refined lactose 20.00g, inject the about 700ml of water and dissolve, extremely suitable with the sulfuric acid regulation solution pH of 5mol/L, then with 2mol/L potassium hydroxide solution, return and adjust pH to suitable, add water for injection to be diluted to 800ml, shake up, use 0.22um filtering with microporous membrane, subpackage, half tamponade, lyophilizing, tamponade, Zha Gai, obtains this product.Shown in table 3 specific as follows.

Table 3: tigecycline injectable powder stability test:

Experimental group	PH value	0 day	40 ℃ one month	40 ℃ two months	40 ℃ three months	40 ℃ six months
							1	4.4 pull back to 5.6	0.221	0.489	0.695	0.871	1.302
2	1.0 pull back to 4.5	0.192	0.360	0.462	0.513	0.558

Embodiment 3:

Get tigecycline 10.03g, refined lactose 20.00g, inject the about 700ml of water and dissolve, extremely suitable with the phosphoric acid regulator solution pH of 5mol/L, then with 2mol/L sodium hydroxide solution, return and adjust pH to suitable, add water for injection to be diluted to 800ml, shake up, use 0.22um filtering with microporous membrane, subpackage, half tamponade, lyophilizing, tamponade, Zha Gai, obtains this product.Shown in table 4 specific as follows.

Table 4: tigecycline injectable powder stability test:

Experimental group	PH value	0 day	40 ℃ one month	40 ℃ two months	40 ℃ three months	40 ℃ six months
							1	1.5 pull back to 5.6	0.210	0.298	0.332	0.358	0.472
2	2.0 pull back to 6.3	0.221	0.353	0.438	0.492	0.735

Claims (6)

1. an injectable powder preparation method that reduces tigecycline epimer, is characterized in that comprising the following steps: get tigecycline, refined lactose, be dissolved in water for injection, first with acid, adjust and make material dissolution and make the pH of solution to suitable, then with alkali, return and adjust pH to suitable, 0.22 μ m filtering with microporous membrane, subpackage, half tamponade, lyophilizing, tamponade, Zha Gai, obtains, wherein, it is that pH is 1.0 to 4.4 that pH scope is adjusted in described acid, and described alkali returns and adjusts pH scope is that pH is 4.5 to 6.3.

2. injectable powder preparation method according to claim 1, is characterized in that: described acid is mineral acid, and described alkali is inorganic base.

3. injectable powder preparation method according to claim 2, is characterized in that: described acid is medicinal hydrochloric acid, sulphuric acid or phosphoric acid.

4. injectable powder preparation method according to claim 2, is characterized in that: described alkali is medicinal sodium hydroxide or potassium hydroxide.

5. injectable powder preparation method according to claim 1, is characterized in that: it is that pH is 1.0 to 1.5 that pH scope is adjusted in described acid.

6. injectable powder preparation method according to claim 1, is characterized in that: described alkali returns and adjusts pH scope is that pH is 5.2 to 6.3.