CN1660407A - Liquid preparation containing penta peptide of thymus and preparing method - Google Patents

Abstract

A liquid preparation containing thymopeptide-5 is disclosed. Its preparing process features that the buffering liquid of citrate is used to regulate pH=6.5-7.5, the sodium chloride is chosen as the optimal osmolarity regulator, and the stabilizer and protector are also used.

Description

Contain liquid preparation of Thymopentin and preparation method thereof

Technical field

The present invention relates to the new liquid preparation of Thymopentin and preparation method thereof that contains.

Background technology

Thymopentin (TP5) is the pentapeptide of synthetic, and its amino acid sequence is identical with the 32-36 amino acids ingredient of thymopoietin (TP) with structure, and promptly Arg-Lys-Asp-Val-Tyr is a TP important function active part.TP5 can promote the differentiation and the growth of thymus and periphery T cell, and body's immunological function is had dual regulation, can make too high or too low immunoreation trend normal.Clinical research proof Thymopentin has important regulatory role to immunologic hypofunction and autoimmune patient's immunologic function.

Thymopentin is mainly used in clinically: the damnification of immunity function due to the malignant tumor; Chronic type b, hepatitis C and complication thereof; Surgery large operation and severe infections; Type ii diabetes; Dermatosis and sexually transmitted disease (STD) are as genital herpes, condyloma acuminatum, herpes zoster etc.; Autoimmune disease is as rheumatoid arthritis, lupus erythematosus; Climacteric syndrome and old immunologic hypofunction; Congenital thymus function defective.

The polypeptide commonly used and the preparation of protein drug are lyophilized preparation, but the lyophilizing preservation method has the shortcoming of himself: freeze-dry process is the cost height not only, and in cryodesiccated process, concentration is excessive, may cause the instability of polypeptide and protein drug.The unstability of polypeptide drugs directly influences patient's treatment, and under the normal dose, the medicine that patient absorbs does not reach treatment concentration.Because degree the unknown of content forfeiture, so simply improve the practical application dosage that method that dosage compensates can not accurately be controlled medicine, the toxic and side effects that may cause the overdose administration to bring simultaneously.Therefore need a kind of stable liquid preparation to guarantee clinical use.The invention provides a kind of new Thymopentin liquid preparation and preparation method and satisfy above-mentioned needs with longer storage time.

Summary of the invention

Through further investigation, the inventor finds, adopts the liquid preparation of citrate buffer preparation Thymopentin, the Thymopentin liquid preparation of low concentration especially, and its stability obviously improves, and (4 ℃) can be stablized 2 years at least under refrigerated condition.Based on this unexpected discovery, finished the present invention.

The present invention contains the liquid preparation of Thymopentin, it is characterized in that containing citrate buffer.

Thymopentin liquid preparation pH value of the present invention is 6.0-8.0, and preferred pH value is 6.5-7.5, and optimum is 7.2.The bronsted lowry acids and bases bronsted lowry of adjusting pH should be physiological safety, and the present invention requires to use HCl and NaOH.

It is osmotic pressure regulator that the present invention selects NaCl for use, and sodium chloride concentration is 80-140mM.

The present invention is the Thymopentin liquid preparation of special buffer, contains the 0.5-50mg/ml Thymopentin.

The liquid preparation that the present invention contains Thymopentin can be prepared by following method, and this method comprises:

(a) in described Thymopentin solution, add the buffer that contains citrate, or change the buffer of described Thymopentin solution into contain citrate buffer;

(b) the Thymopentin solution of packing step a acquisition.

The method according to this invention, Thymopentin liquid preparation adopt the citrate buffer preparation.This buffer has been applied to multiple medicine, extensively in clinical use, can think that physiology is compatible and safety non-toxic.Although many buffer also have above characteristic, as acetate buffer, Hepes buffer, phosphate buffer, succinate buffer etc., what it is considered herein that optimum is citrate buffer, and citric acid salt concentration is 10-50mM.

Description of drawings

Fig. 1 is a bar diagram, be presented in the accelerated test, Thymopentin in various buffer 45 ℃ preserve after 10 days the percent of assay.The used buffer of sample A is the 30mM acetate buffer, 5% mannitol, pH7.2; The used buffer of sample P is the 30mM phosphate buffer, 5% mannitol, pH7.2; The used buffer of sample C is the 30mM citrate buffer, 5% mannitol, pH7.2; The used buffer of sample M is 5% mannitol, pH7.2.

Fig. 2 is a line diagram, be presented in the long term test, Thymopentin in containing the citrate buffer of sodium chloride 4 ℃ preserved 24 months, respectively at the percent of 0th month, 3 months, 6 months, 9 months, 12 months, 18 months, 24 months Thymopentin assays.

The specific embodiment

Elaborate the preparation technology and the stability thereof of Thymopentin liquid preparation below for example.These examples only are used for explanation, do not limit range of application of the present invention.

Example 1, buffer are to the effect of Thymopentin stability

Press the listed buffer preparation of table 1 Thymopentin solution respectively, concentration is 1mg/ml.The injection that 1ml/ props up is made in the filtering with microporous membrane degerming of gained solution.Sample is put into 45 ℃ of calorstats, respectively the 0th day sample and the 10th day sample are measured content as follows.

The buffer that each sample of table 1 is used

Sample	Buffer
		????A	The 30mM acetate buffer, 5% mannitol, pH7.2
????P	The 30mM phosphate buffer, 5% mannitol, pH7.2
		????C	The 30mM citrate buffer, 5% mannitol, pH7.2
????M	5% mannitol, pH7.2

Content assaying method adopts high performance liquid chromatography, is filler with the octadecyl silane, and phosphate buffer-methanol of pH7.0 (60: 40) is mobile phase; The detection wavelength is 275nm.Theoretical cam curve is calculated by the Thymopentin peak should be not less than 1200.Accurately measure reference substance solution and sample solution sample introduction, press external standard method with calculated by peak area Thymopentin content.

In the 0th day sample size was 100%, calculated each sample and quickened the 10th day assay percent.Fig. 1 shows the stable contrast and experiment that each buffer sample was preserved 10 days under 45 ℃ of conditions.Show that Thymopentin is being that stability is optimum in the solvent with the citrate buffer.

Example 2, contain of the effect of the citrate buffer of sodium chloride to Thymopentin stability

For further investigating the citrate buffer that contains sodium chloride to Thymopentin stability of (4 ℃) under refrigerated condition, prepared 1mg/ml Thymopentin liquid preparation (pH7.2), wherein contain the citrate buffer of 30mM and the sodium chloride of 118mM, NaCl is an osmotic pressure regulator.Be stored in 4 ℃ and observed 24 months, pressed content assaying method respectively at 0th month, 3 months, 6 months, 9 months, 12 months, 18 months, 24 months and measure Thymopentin content.

In 0th month sample content was 100%, and calculation sample is at each time point assay percent respectively.Fig. 2 show sample is 24 months stability test results under 4 ℃ of conditions.The result shows: under 4 ℃ of conditions, Thymopentin content in containing the citrate buffer solution of sodium chloride does not have significance to be changed, and can stablize at least 2 years.

Example 3, contain the preparation of the liquid preparation of citrate buffer

Prescription: Thymopentin 1.0g

Sodium citrate 6.9g

Sodium chloride 8.8g

Water for injection adds to 1000ml

Operating procedure: get about 600ml water for injection and add in the proper container, it is clear and bright that adding 6.9g sodium chloride and 8.8g sodium citrate are stirred to solution.Stirring (preventing to generate foam) extremely all dissolvings and clear and bright of adding 1.0g Thymopentin down subsequently.Gained solution is transferred pH7.0-7.4 with 0.1M NaOH or 0.1MHCl.Add the injection water to 1000ml, be stirred to clear and bright fully.Gained solution is considered the membrane filtration degerming with micropore, and the injection that 1ml/ props up is made in packing.

Claims (10)

1. the liquid preparation that contains Thymopentin is characterized in that containing citrate buffer.

2. the liquid preparation of claim 1 is characterized in that pH value is 6.0-8.0.

3. the liquid preparation of claim 1 is characterized in that pH value is 6.5-7.5.

4. the liquid preparation of claim 1 is characterized in that pH value is 7.2.

5. claim 1,2 or 3 liquid preparation is characterized in that further containing sodium chloride.

6. the liquid preparation of claim 5 is the human injection.

7. the liquid preparation of claim 6, wherein the concentration of Thymopentin is 0.5-50mg/ml.

8. the liquid preparation of claim 6, wherein citric acid salt concentration is 10-50mM.

9. the liquid preparation of claim 6, wherein sodium chloride concentration is 80-140mM.

10. preparation contains the method for Thymopentin liquid preparation, comprising:

(a) in described Thymopentin solution, add the buffer that contains citrate, or change the buffer of described Thymopentin solution into contain citrate buffer;

(b) the Thymopentin solution of packing step a acquisition.