CN103768584A - Relaxin injection - Google Patents

Abstract

The invention discloses a relaxin injection preparation, which contains relaxin, citrate or acetate buffer solution, NaCl, PEG200 (polyethylene glycol 200) or PEG400 and bovine serum albumin or human serum albumin, wherein the concentration of relaxin is at least 1.5 mg/mL; the pH range is 4.5-5.5; the ion strength is 0.1-0.2 mu m; the concentration of the PEG200 or PEG400 is 5-15% (W/V); the concentration of bovine serum albumin or human serum albumin is 0.01-5% (W/V). Researches show that the relaxin injection disclosed by the invention is capable of keeping the stability of high-concentration relaxin in a water solution, solving the precipitation accumulation problem of injection caused by increased concentration of relaxin, being beneficial to realizing high-concentration micro-injection of the relaxin injection, and being convenient for patients to use.

Description

A kind of Cervilaxin injection

Technical field

The present invention relates to a kind of pharmaceutical preparation of Cervilaxin, particularly a kind of injection formulation of Cervilaxin.

Background technology

Cervilaxin, English Relaxin by name, medicament universal is called Serelaxin, is that a kind of gene recombinaton or chemical synthesis process of adopting prepared RLN2.Its structure is made up of two peptide chains, and A chain is containing 24 aminoacid and have intrachain disulfide bond, and B chain is containing 29 aminoacid, and A chain is connected by two disulfide bond with B chain, and its structural formula is as follows:

Wherein pGlu is pyroglutamic acid, and Tyr is tyrosine, and Ser is serine, and Asn is agedoite, Cys is cysteine, and His is histidine, and Val is valine, and Gly is glycine, Thr is threonine, and Lys is lysine, and Arg is arginine, Leu is leucine, and Phe is phenylalanine, and Asp is aspartic acid, Trp is tryptophan, and Met is methionine, and Glu is glutamic acid, Ile is isoleucine, and Ala is alanine, and Gln is glutamine.

Research in recent years finds, Cervilaxin has significant therapeutic effect to heart failure tool, can blood vessel dilating, increase arterial compliance, the dyspnea symptom of significantly alleviating heart failure and causing.U.S. Pat 5166191 discloses Cervilaxin and has been used for the treatment of the especially process patent of heart failure of cardiovascular disease, Chinese patent literature CN1052655C(application number 89100947.7, applying date 1989.02.25) disclose pH value of solution and the impact of ionic strength on Cervilaxin stability, and a kind of liquid preparation of Cervilaxin disclosed, the suitable concentration of this liquid preparation is about 1.0 mg/mL, but fails to solve the significantly reduced problem of preparation stability that Cervilaxin injection Enrichment causes.Chinese patent literature CN102133200 B(application number 201110062557.4, applying date 2011.03.16) a kind of freeze-dried powder preparation and preparation technology thereof of Cervilaxin disclosed.But freeze-dried powder preparation needs redissolution operation before application, thereby incur loss through delay the best occasion for the treatment of acute heart failure.

Clinical research shows, the dosage that Cervilaxin is used for the treatment of heart failure is approximately 30 μ g/kg/ days～250 μ g/kg/ days, according to conditions of patients needs, dosage may be incremented to 960 μ g/kg/ days, need to adopt 24 hours～72 hours～the mode administration of venoclysis continuously.Low concentration Cervilaxin injection increases administration volume, brings great inconvenience to patient and clinical nursing activity.Adopt the micro-injection of high concentration Cervilaxin, can significantly reduce administration volume, be suitable for being assembled in medicine micro-injection pump, injection process does not affect patient's feed and freely movable.But research is found (to see the research paper " Stability and Characterization of Recombinant Human Relaxin " of Tue H. Nguyen and Steven J. Shire, this paper publishing is in Formulation, Characterization, and Stability of Protein Drugs:Case Histories. Pharmaceutical Biotechnology, the 9th volume in 2002, 247-274 page), Cervilaxin is the configurable the highest 1.0mg/mL that only can reach of concentration of aqueous solution with physical stability in the scope of pH3.0～9.0, concentration higher than 1.0mg/mL after, long-term storage meeting crystallization or precipitation are assembled, adopt Chinese patent literature CN1052655C(application number 89100947.7, applying date 1989.02.25) technical scheme cannot effectively solve the Cervilaxin injection concentration significantly reduced problem of stability causing that raises.And only adopt conventional pharmaceutic adjuvant to make high concentration Cervilaxin injection reach desirable stability as the conventional preparation treatment technologies of employing such as buffer agent, surfactant, sugar and polyhydric alcohol still cannot reach.

In view of the foregoing, research one has ideal stability, can extend the holding time, containing the injection of high concentration Cervilaxin, has important practical significance.

Summary of the invention

For above-mentioned prior art, the object of this invention is to provide one and contain high concentration Cervilaxin, and can improve the ejection preparation of Cervilaxin physical stability.

For achieving the above object, the technical solution used in the present invention is:

A kind of Cervilaxin injection, contains following composition:

1) Cervilaxin of 1.5mg/mL at least;

2) being suitable for maintaining pH scope is 4.5～5.5 citrate or acetate buffer solution;

3) be suitable for maintaining the NaCl that ionic strength is 0.1 μ～0.2 μ;

4) PEG200 of 5%～15% (W/V) or PEG400;

5) bovine serum albumin of 0.01%～5% (W/V) or human serum albumin.

As further optimization of the present invention, described PEG200 or PEG400 content are 6% (W/V).

As further optimization of the present invention, described bovine serum albumin or human serum albumin's content is 2% (W/V).

Another aspect of the present invention, provides a kind of preparation method of Cervilaxin injection, comprises the following steps:

1) water for injection being regulated to pH with citrate or acetate buffer solution is 4.5～5.5, adds NaCl, and making solution ion strength is 0.1 μ～0.2 μ;

2) Cervilaxin is dissolved in the solution of step 1), add PEG200 or PEG400, be uniformly mixed, 0 ℃～5 ℃ are stirred placement 12 hours～24 hours, then add bovine serum albumin or human serum albumin, regulating pH with citrate or acetate buffer solution is 4.5～5.5; Described PEG200 or the PEG400 concentration in solution is 5%～15% (W/V); Described bovine serum albumin or the human serum albumin concentration in solution is 0.01%～5% (W/V).

The present inventor is unexpected discovery after often a large amount of preparation prescription screenings, a small amount of albumin comprises that bovine serum albumin and human serum albumin and Polyethylene Glycol comprise PEG200 and PEG400, be used in conjunction with according to finite concentration ratio, can improve significantly the stability of high concentration Cervilaxin injection, extend the pot-life.Accelerated stability test result shows, Cervilaxin injection of the present invention, having good stability, is not assembled, precipitated in accelerated test condition.Preparation method that the present invention adopts, process conditions are easy to control, are suitable for large-scale industrial production.

The specific embodiment

Below in conjunction with embodiment, the present invention is further explained.Should be understood that, following examples are only for explaining the present invention, rather than limit the scope of the invention.

The preparation (not containing PEG200 or PEG400) of reference embodiment 1 Cervilaxin injection

1) 100mL water for injection being regulated to pH with citrate buffer solution is 5.0, adds NaCl, and making solution ion strength is 0.15 μ;

2) Cervilaxin 150mg is dissolved in the solution of step 1), adds mannitol 35mg, then adding bovine serum albumin to make its concentration is 2%(W/V), stir 3 ℃ to stir and place 20 hours, regulating pH with citrate buffer solution is 5.0.

The preparation (not containing bovine serum albumin or human serum albumin) of reference embodiment 2 Cervilaxin injection

1) 100mL water for injection being regulated to pH with acetate buffer solution is 5.5, adds NaCl, and making solution ion strength is 0.18 μ;

2) Cervilaxin 150mg is dissolved in the solution of step 1), adding PEG400 to make its concentration is 6%(W/V), be uniformly mixed, 5 ℃ are stirred placement 24 hours, and regulating pH with acetate buffer solution is 5.5.

The preparation (not containing PEG200, PEG400, bovine serum albumin, human serum albumin) of reference embodiment 3 Cervilaxin injection

1) 100mL water for injection being regulated to pH with citrate buffer solution is 5.5, adds NaCl, and making solution ion strength is 0.15 μ;

2) Cervilaxin 200mg is dissolved in the solution of step 1), adds sorbitol 50mg, be uniformly mixed, 2 ℃ are stirred placement 12 hours, and regulating pH with citrate buffer solution is 5.5.

The preparation of embodiment 1 Cervilaxin injection

1) 100mL water for injection being regulated to pH with citrate buffer solution is 5.0, adds NaCl, and making solution ion strength is 0.15 μ;

2) Cervilaxin 250mg is dissolved in the solution of step 1), adding PEG200 to make its concentration is 10%(W/V), be uniformly mixed 2 ℃ and stir placement 15 hours, then adding bovine serum albumin to make its concentration is 2%(W/V), regulating pH with citrate buffer solution is 5.0.

The preparation of embodiment 2 Cervilaxin injection

1) 100mL water for injection being regulated to pH with acetate buffer solution is 5.2, adds NaCl, and making solution ion strength is 0.18 μ;

2) Cervilaxin 300mg is dissolved in the solution of step 1), adding PEG400 to make its concentration is 6%(W/V), be uniformly mixed 3 ℃ and stir placement 18 hours, then adding human serum albumin to make its concentration is 0.1%(W/V), regulating pH with acetate buffer solution is 4.8.

The preparation of embodiment 3 Cervilaxin injection

1) 100mL water for injection being regulated to pH with acetate buffer solution is 5.0, adds NaCl, and making solution ion strength is 0.15 μ;

2) Cervilaxin 350g is dissolved in the solution of step 1), adding PEG400 to make its concentration is 6%(W/V), be uniformly mixed 2 ℃ and stir placement 20 hours, then adding bovine serum albumin to make its concentration is 2%(W/V), regulating pH with acetate buffer solution is 5.0.

The preparation of embodiment 4 Cervilaxin injection

1) 100mL water for injection being regulated to pH with citrate buffer solution is 5.5, adds NaCl, and making solution ion strength is 0.15 μ;

2) Cervilaxin 400mg is dissolved in the solution of step 1), adding PEG200 to make its concentration is 12%(W/V), be uniformly mixed 2 ℃ and stir placement 20 hours, then adding human serum albumin to make its concentration is 1.0%(W/V), regulating pH with citrate buffer solution is 5.5.

Embodiment 5 stability tests

Get reference embodiment 1 and be labeled as respectively reference 1 to reference 3 to reference embodiment 3() and each 10 equal portions of Cervilaxin injection prepared of embodiment 1 to embodiment 4, every part of 10ml, is placed in centrifuge, at 37 ℃, and centrifugal 5 hours of 4000rmp.Observe and record each group of sample occur precipitation time (minute), centrifugal end precipitation separation weighs precipitation weight, the percent that levels of precipitate accounts for Cervilaxin content in sample with precipitation weight represents, it is poor that precipitation occurs that very fast or final levels of precipitate is considered as more greatly preparation stability, the results are shown in following table:

?	Reference 1	Reference 2	Reference 3	Embodiment 1	Embodiment 2	Embodiment 3	Embodiment 4
								Sedimentation time (minute)	92	108	115	189	227	165	194
Levels of precipitate (%)	5.7	12.2	9.5	2.9	1.6	1.0	3.4

From above table, the high concentration relaxin injection that contains PEG200 or PEG400 and bovine serum albumin or human serum albumin, under centrifugal condition, the time that occurs precipitation is significantly later than the injection that does not contain above-mentioned component or only contain above-mentioned a kind of component, and in concentration higher than reference embodiment Cervilaxin solution in the situation that, the Cervilaxin solution that after centrifugal 5 hours, total levels of precipitate is still significantly prepared lower than reference embodiment.

Claims (4)

1. a Cervilaxin injection, is characterized in that, contains following composition:

1) Cervilaxin of 1.5mg/mL at least;

2) being suitable for maintaining pH scope is 4.5～5.5 citrate or acetate buffer solution;

3) be suitable for maintaining the NaCl that ionic strength is 0.1 μ～0.2 μ;

4) PEG200 of 5%～15% (W/V) or PEG400;

5) bovine serum albumin of 0.01%～5% (W/V) or human serum albumin.

2. Cervilaxin injection according to claim 1, is characterized in that, described PEG200 or PEG400 content are 6% (W/V).

3. Cervilaxin injection according to claim 1, is characterized in that, described bovine serum albumin or human serum albumin's content is 2% (W/V).

4. the preparation method of the Cervilaxin injection described in any one in claims 1 to 3, is characterized in that, comprises the following steps:

1) water for injection being regulated to pH with citrate or acetate buffer solution is 4.5～5.5, adds NaCl, and making solution ion strength is 0.1 μ～0.2 μ;

2) Cervilaxin is dissolved in the solution of step 1), add PEG200 or PEG400, be uniformly mixed, 0 ℃～5 ℃ are stirred placement 12 hours～24 hours, then add bovine serum albumin or human serum albumin, regulating pH with citrate or acetate buffer solution is 4.5～5.5; Described PEG200 or the PEG400 concentration in solution is 5%～15% (W/V); Described bovine serum albumin or the human serum albumin concentration in solution is 0.01%～5% (W/V).