CN1272069C - Stabilizing medicinal agent containing ciliary nerve nutritive factor analogue - Google Patents

Abstract

The present invention discloses a stabilizing medicinal agent containing ciliary nerve nutritive factor analogues. Except for using the ciliary neurotrophic factor (CNTF) analogs as an effective component, the medicament also at least comprises a substance selected from medicinal surfactants, protein stabilizing agents and inorganic salt, and the effective components are stably stored for a long time by using the form of a water solution or a freeze-dried preparation. The present invention has the advantage that the present invention prepares a ciliary neurotrophic factor analog preparation which is suitable for clinical application, particularly for injection and stable storage by adding components which can be accepted by human bodies. The preparation can prevent active components (ciliary neurotrophic factor analogs) from being lost or devitalized by multiple factors such as container absorption, protein denaturation, polymerization, degradation, oxidation, etc.; thereby, the preparation is convenient for transportation, long-stem storage and clinical application.

Description

Stabilized agents containing ciliary neurotrophic factor analogs

Technical field

The present invention relates to a stable medicine containing ciliary neurotrophic factor analogs. More specifically, the present invention relates to a stable preparation containing ciliary neurotrophic factor analogs, suitable for subcutaneous or intravenous administration, for the treatment of neurological diseases or obesity and related diseases.

Background technique

CNTF was first discovered in vitro to promote the survival of chick ciliary ganglion (Manthorpe et al., 1980, J.Neurochem.34: 69-75), and in the following decades, many Related biological functions were discovered. Hughes et al. found that it can promote the differentiation of glial progenitor cells in the visual and cranial nerves of perinatal mice (Hughes et al., 1988, Nature 335:70-73), and also found that it can promote the differentiation of dorsal root ganglia in chick embryos. Growth (Skaper and Varon, 1986, Brain Res. 389:39-46). In addition, he can also promote the growth and differentiation of motor neurons, hippocampal neurons and parasympathetic spinal cord neurons (Ip, et al.1991, J.Neurosci.11: 3124-3134; Blottner, et al.1989, Neurosci.Lett .105:316-320).

In view of the multiple biological functions of CNTF, it is believed that it can play a certain role in repairing nerve damage and treating neurotrophic diseases, so it has a good prospect for clinical application. In further studies, it was also found that CNTF can also play a role in the treatment of obesity, especially Leptin-resistant obesity, showing possible clinical applications (Isabelle Gloaguen, et al. 1997, Proceedings of the National Academy of Sciences of the United States of America). In addition, further studies have also shown that the biological activity and stability of CNTF analogues obtained after mutation through genetic engineering methods have been improved. However, as a protein drug, its stability still cannot be compared with conventional chemical drugs (Panayotatos; Nikos, 1998, US 5846935). Therefore, it is extremely meaningful to develop a formulation that can stably preserve ciliary neurotrophic factor analogues and is suitable for actual clinical use. What the present invention relates to is exactly this aspect content.

Contents of the invention

The object of the present invention is to propose a stable agent containing ciliary neurotrophic factor analogs.

In addition to ciliary neurotrophic factor (CNTF) analogs as active ingredients, at least one is selected from pharmaceutically acceptable surfactants, protein stabilizers, inorganic salts, and its solution pH or The pH of the solution was between 5-8 after reconstitution.

The advantage of the present invention is to prepare a ciliary neurotrophic factor analog preparation suitable for clinical use, especially for injection and stable storage by adding some components acceptable to the human body. This preparation can prevent the loss or inactivation of active ingredients (ciliary neurotrophic factor analogs) due to container adsorption, or protein denaturation, aggregation, degradation, oxidation and other factors, thus facilitating transportation, long-term storage and clinical use.

Detailed ways

The ciliary neurotrophic factor analogs involved in the present invention are recombinant products prepared by genetic engineering methods, and its preparation method can be obtained from U.S. Patent No. 5,846,935, and can also be carried out according to a patent submitted by the same applicant of the present invention (Chinese Patent Application number: 02136033.2). The protein sequence of the obtained product is inconsistent with natural human CNTF. Any highly purified ciliary neurotrophic factor analogue can be added to the medicament of the present invention as the main active ingredient, and the best one is the product obtained by separation and purification after genetic engineering and expression by Escherichia coli. The amino acid structures of several CNTF analogs most suitable for the present invention are indicated below:

Ciliary neurotrophic factor analog analog sequence 1: 9 19 29 39 49 AFTE HSPLT PHRRDLASRS IWLARKIRSD LTALTESYVK HQGLNKNINL 59 69 79 89 99 DSADGMPVAS TDRWSELTEA ERLQENLQAY RTFHVLLARL LEDQQVHFTP 109 119 129 139 149 TEGDFHQAIH TLLLQVAAFA YQIEELMILL EYKIPRNEAD GMPINVGDGG 159 169 179 LFEKKLWGLK VLQELSQWTV RSIHDLRFIS SHQTG

Ciliary neurotrophic factor analog analog sequence 2: 10 20 30 40 50 MAFTE HSPLT PHRRDLASRS IWLARKIRSD LTALTESYVK HQGLNKNINL 60 70 80 90 100 DSADGMPVAS TDRWSELTEA ERLQENLQAY RTFHVLLARL LEDQQVHFTP 110 120 130 140 150 TEGDFHQAIH TLLLQVAAFA YQIEELMILL EYKIPRNEAD GMPINVGDGG 160 170 180 LFEKKLWGLK VLQELSQWTV RSIHDLRFIS SHQTG

Ciliary neurotrophic factor analog analog sequence 3: 10 20 30 40 50 MAFTE HSPLT PHRRDLCSRS IWLARKIRSD LTALTESYVK HQGLNKNINL 60 70 80 90 100 DSADGMPVAS TDRWSELTEA ERLQENLQAY RTFHVLLARL LEDQQVHFTP 110 120 130 140 150 TEGDFHQAIH TLLLQVAAFA YQIEELMILL EYKIPRNEAD GMPINVGDGG 160 170 180 LFEKKLWGLK VLQELSQWTV RSIHDLRFIS SHQTGIP

But the present invention is not limited to CNTF analogs of these structures. Moreover, the method provided in this patent is also applicable to other CNTF analogues obtained through genetic engineering methods for similar protein mutations. Because these changes are considered by professionals to be extremely easy to implement. Therefore the application in this respect is protected by this patent.

At present, the dose of ciliary neurotrophic factor analogues used clinically for the treatment of obesity is extremely small, generally 1-500 micrograms (preferably 20-200 micrograms) at a time, and used continuously for more than one week (preferably more than 10 weeks). ). Because the administration takes a long time, it is generally taken home by the patient for self-use.

However, ciliary neurotrophic factor analogs are not stable as proteins themselves. Although they have been improved after transformation (Panayotatos; Nikos, 1998, US 5846935), their activity will still be affected by various factors during long-term storage. The influence of environmental factors. Such as temperature, humidity, oxygen and UV are highly sensitive. Due to the action of these factors, various physical or chemical changes such as binding, polymerization, and oxidation may occur and activity is largely lost. Therefore, if the stability of ciliary neurotrophic factor analogs cannot be guaranteed during storage, it will lead to changes in the dosage and thus affect the curative effect.

Therefore, it is extremely necessary to design a stable and easy-to-preserve ciliary neurotrophic factor analog preparation, which can fully prevent the activity of its active components from decreasing. This is the main objective of the present invention.

In order to improve the stability of the medicament containing ciliary neurotrophic factor analogs, the present inventors have carried out in-depth research and found that if from pharmaceutically acceptable surfactants, suitable protein stabilizers (such as sugars, proteins, amino acids and derivatives thereof) This goal can be effectively achieved by selecting at least one of substances, antioxidants and macromolecular compounds) and inorganic salts to be added to the preparation.

Surfactants, protein stabilizers, and inorganic salts mentioned in the present invention may be used in combination with only one of them or in combination. On the basis of the present invention, changing its concentration or adding some other substances, but the changes that have no significant effect on improving the stability of ciliary neurotrophic factor analogues are still regarded as a part of the present invention.

Typical examples of surfactants suitable for use in the present invention are described below:

Nonionic surfactants with 6-13 HLB, such as sorbitan fatty acid esters, glycerin fatty acid esters (such as sorbitan caprylic monoester, sorbitan lauric monoester and sorbitan palmitic monoester ), polyglyceryl fatty acid esters (such as glyceryl caprylic monoester, glyceryl myristic monofatty acid monoester and glyceryl stearic fatty acid monoester), polyoxyethylene sorbitan fatty acid ester, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene Ethylene glycerin fatty acid ester, polyoxyethylene glycol fatty acid ester, polyoxyethylene alkyl ether, polyoxyethylene polyoxypropylene alkyl ether, polyoxyethylene phenyl ether, polyoxyethylated hard castor oil, polyoxyethylene Ethylated beeswax derivatives, polyoxyethylated lanolin derivatives or polyoxyethylene fatty acid amides, cationic surfactants are alkyl sulfates (such as those with a C ₁₀ -C ₁₈ alkyl group), poly Vinyl alkyl ether sulfates, sulfosuccinates, natural surfactants are: lecithin, glycerophospholipids, sphingomyelin, sucrose fatty acid esters, etc. Of course, these surfactants can be used alone or in combination.

The added amount of the surfactant is preferably between 0.01-1000 by weight to the ciliary neurotrophic factor analogue.

The protein stabilizer used in the preparation of the present invention should be acceptable to the human body, and can be carbohydrates, proteins, amino acids and their derivatives, and macromolecular compounds.

The sugars therein can be selected from a variety of monosaccharides, oligosaccharides and polysaccharides, as well as phospholipids and nucleotide derivatives. Typical examples are as follows: trivalent and higher sucrose alcohols such as glycerol, erythritol, arabitol, xylitol, sorbitol and mannitol; acidic sugars such as glucuronic acid, iduronic acid, ceramide Sugar acid, galacturonic acid, glucuronic acid, mannuronic acid, hyaluronic acid and its salts, chondroitin sulfate and its salts, also heparin, inulin, chitin and its derivatives, dextrin, average Dextran and alginic acid and their salts with a molecular weight of 5,000-1,50,000. These sugars may be added alone or in combination.

The added amount of sugar is preferably between 10-5000 by weight to ciliary neurotrophic factor analogue.

Proteins suitable for the present invention may include human serum albumin, human serum albumin, human serum globulin, gelatin, acid- or alkali-treated gelatin with an average molecular weight of 5,000-10,000, and collagen. In order to avoid blood-borne contamination, these proteins may be extracted naturally, or may be mutants prepared or obtained by means of genetic engineering or other methods. These proteins can be used either singly or in combination.

The contained protein quantity is preferably between 1-5000 by weight ratio to ciliary neurotrophic factor analogue.

Polypeptides, amino acids and derivatives suitable for the present invention can be selected from the following group of substances: glutathione, glycine, alanine, serine, aspartic acid, glutamic acid, threonine , tryptophan, lysine, hydroxylysine, histidine, arginine, cystine, cysteine, methionine, phenylalanine, leucine, isoleucine, etc. and their derivatives. The added amount is between 10-5000 in weight ratio to ciliary neurotrophic factor analogue.

Typical examples of high molecular compounds suitable for use in the present invention include natural polymers such as hydroxypropyl cellulose, hydroxymethyl cellulose, carboxymethyl cellulose and hydroxyethyl cellulose, synthetic polymers such as polyethylene glycol Acids (molecular weight 300-6,000), polyvinyl alcohol (molecular weight 400-100,000) and polyvinylpyrrolidone (molecular weight 20,000-100,000). Of course, these polymer compounds may be used alone or in combination.

The weight ratio of the high molecular weight compound to the ciliary neurotrophic factor analog is preferably between 1-5000.

In addition to the protective agents of the above-mentioned surfactants, sugars, proteins, polypeptides and amino acids or polymer compounds, at least one of the sulfur-containing reducing agents and antioxidants can be selected to be added to the present invention containing ciliary neurotrophic factors. in medicines. The sulfur-containing reducing agent referred to includes lipoic acid, thiodiglycol, thioethanolamine, thiosulfate, sodium sulfenate, sodium metabisulfite, sodium sulfite, dithiothreitol, and the like. Antioxidants mentioned include: erythorbic acid, butylated hydroxytoluene, tocopheryl acetate, L-ascorbic acid and its salts, l-ascorbyl palmitate. L-Ascorbyl stearate, etc.

The added amount of the sulfur-containing reducing agent and the antioxidant is between 0.001-100 in weight ratio to the ciliary neurotrophic factor analogue.

In the medicament combination of the present invention, various inorganic salts can also be added to regulate osmotic pressure, maintain pH, and complex metal ions. Such as adding sodium chloride, potassium chloride, urea, phosphate, acetate, citrate, barbiturate, tris, borate, succinate, EDTA, etc. The added amount is between 0.01-1000 in weight ratio to ciliary neurotrophic factor analog.

In order to prepare the stable preparation containing ciliary neurotrophic factor analogs of the present invention in a suitable dosage form, one or more of the following reagents may also be added: diluent, co-solvent, etc., the most suitable is water for injection. If it is prepared in the form of an aqueous solution, a certain amount of preservatives, such as benzyl alcohol, chlorobutanol, parabens, etc., can also be added to prevent microbial contamination.

The stable formulation of the invention is most suitable for parenteral administration, such as subcutaneous or intravenous injection. Its final preparation form can be aqueous injection or freeze-dried preparation. The stability of the lyophilized agent is better than the former.

One of the above-mentioned medicament compositions can be selected to be added, but it is better to select several types of substances to be added in combination, so as to better achieve the purpose of the present invention. For example: at least one of surfactants, sugars, proteins or polymer compounds can be added, which can prevent the ciliary neurotrophic factor analogs from being adsorbed on the container wall or syringe, and can be used for a long time Keep ciliary neurotrophic factor analog stable. The detailed mechanism for stabilizing ciliary neurotrophic factor analogues and preventing adsorption using the above substances has been partially elucidated. In the presence of a surfactant, its hydrophobic part can be combined with the ciliary neurotrophic factor analog to cover it, so that it can prevent the ciliary neurotrophic factor analog from being adsorbed on the container wall and the syringe, Also prevents the formation of ciliary neurotrophin analog aggregates. Sugars or hydrophilic macromolecular compounds form a hydration layer between ciliary neurotrophic factor analogs and the container and syringe walls, so they can effectively prevent adsorption; proteins compete with ciliary neurotrophic factor analogs for adsorption on containers and syringes On the wall, this can effectively inhibit the adsorption of ciliary neurotrophic factor analogues. Amino acids and peptides may act through a mechanism similar to sugars, and sulfur-containing reducing agents and antioxidants can protect the side chains of ciliary neurotrophic factor analogs from oxidation and maintain their biological activity.

Inorganic salts mainly play three roles. First, it increases the ionic strength of the solution, which can prevent denaturation of the ciliary neurotrophic factor analogue and prevent it from being adsorbed to the vessel wall. Second, the preparation can be easily accepted by the human body by adjusting the appropriate osmotic pressure. Third, it can maintain a suitable pH environment, which is conducive to the stability of proteins. Our research found that ciliary neurotrophic factor analogs have better stability between pH5-8.

By adding at least one substance selected from surfactants, protein stabilizers, and inorganic salts, ciliary neurotrophic factor analogs are highly stable and can maintain activity for a long time, which will be confirmed in the following examples. In order to achieve this purpose, the amount of each of these substances, especially the amount of the surfactant protein stabilizer, is critical, and the following range is ideal: Add 0.01 per part of ciliary neurotrophic factor analogue by weight ratio -1000 parts of surfactant, 10-5000 parts of sugar, 1-5000 parts of protein, 1-5000 parts of high molecular compound, 10-5000 parts of amino acid and its derivatives or peptides. In addition, inorganic salts can also be added, so that the final osmotic pressure of the preparation is 250-500 mOsm, and the pH is between 4-9, and the optimum is between 5-8.

In order to further illustrate the present invention, the following examples are provided for the purpose of further illustrating the present invention only and are not meant to be limiting. In these instances, the remaining activity of the ciliary neurotrophin analog can be determined using any of the methods described below.

A) MTT method:

instrument

Carbon dioxide incubator

Clean bench

centrifuge

Enzyme standard colorimeter

MM-miniature oscillator

Reagent

(1) Basic culture medium: RPMI1640 (GIBCOBRL company) plus 10% fetal serum, 0.4mg/ml G-418

(2) Complete culture solution: GM-CSF from GIBCOBRL Company was added to the basic culture solution to 2 ng/ml.

(3) MTT solution: MTT (Fluka Company) was made into a 5.0 mg/ml solution with phosphate buffer. Use a 0.22um filter membrane to filter and sterilize, and store at 4.C in the dark.

(4) Solution: containing 0.01N HCl, 10% SDS aqueous solution.

(5) Cell line: TF-1CNTF-dependent cell line transfected with CNTF receptor α chain, cultured with complete medium at 37°C and 5% CO ₂ , subcultured at intervals of 48-72hr, and controlled cell concentration at 1× Between 10 ^5-6 cells/ml.

operate

(1) The reference substance and the sample to be tested are diluted to 2.0 μg/ml with the basic culture medium according to the protein content, and each step of dilution does not exceed 10 times.

(2) Collect TF-1.CN5a.1 cells by centrifugation. Wash twice with the basic culture medium, centrifuge at 1000 rpm for 5 minutes, resuspend in the basic culture medium, and adjust the cell concentration to 2×10 ⁵ cells/ml.

(3) Prepare ciliary neurotrophic factor analog reference substance and sample double dilution gradient from 1 μg/ml to low concentration on a 96-well plate, 12 gradients for each sample, and 2 or 3 duplicate holes for each dilution , 50 μl per well, and at the same time make blank control, negative control and positive control (containing GM-CSF 4 μg/ml) each at least 3 wells.

(4) Add 50 μl of cell suspension to each well, and culture at 37° C., 5% CO ₂ , and saturated humidity for 72 hours. Add 20 μl/well of MTT solution to the cells 4 hours before the termination of culture.

(4) After the culture was terminated, 100 μl/well of dissolving solution was added, overnight at 37°C, and after the crystals were completely dissolved, the blank control was used to adjust to zero, and the OD _570nm was measured with a microplate reader, and the reference wavelength was 630nm.

result calculation

(1) Take the OD value as the Y axis, and the logarithm of the dilution gradient on the plate as the X axis, and make a scatter diagram of the dose-effect relationship between the reference substance and the sample to be tested. The regression analysis of the experimental results was carried out by Excel software, and the regression equation and correlation coefficient of each sample were obtained. According to the regression equation, the ED ₅₀ value of each sample was calculated.

(2) If the following criteria are met, the experiment is established, otherwise it should be retried: there is no significant difference between the mean value of the positive control and the maximum OD value of each sample, there is no significant difference between the mean value of the negative control and the minimum OD value of each sample, and there is no significant difference between the mean value of the negative control and the minimum OD value of each sample. There was no significant difference among the regression coefficients, and the correlation coefficient of each sample was not less than 0.90.

(3) Calculate the result according to the following formula:

Specific activity of the sample to be tested (U/mg) = specific activity of the reference substance (U/mg) × ED ₅₀ of the reference substance / ED ₅₀ of the sample to be tested

The proportion of remaining ciliary neurotrophic factor analogue activity was calculated according to the following formula:

B) reversed-phase high-performance liquid chromatography:

Octadecylsilane bonded silica gel is used as filler; phase A: trifluoroacetic acid-water solution (take 1.0ml trifluoroacetic acid and add 999ml water, shake well), phase B: trifluoroacetic acid-acetonitrile solution (take 1.0ml three Fluoroacetic acid plus acetonitrile (999ml, shake well) is the mobile phase, gradient elution, the ratio of B phase 2 and 40 minutes is 30% and 70% successively, the detection wavelength is 214nm, and the number of theoretical plates must not be less than 2000.

The proportion of remaining ciliary neurotrophic factor analogue activity was calculated according to the following formula:

There is a good correlation between the remaining ciliary neurotrophic factor analogues measured by the two methods.

Example 1

The various stabilizers listed in Table 1 are added to the ciliary neurotrophic factor analog (its amino acid composition is the same as sequence 1 in the accompanying drawing) of 0.2 mg/ml, containing 10mM pH7.0 phosphate buffer saline in the mixture, without After bacterial filtration, 0.5 ml/cartridge is sealed in a glass vial. Observe its changes under different conditions. Measured by method (A), the results are shown in Table 1. Wherein "activity (%)" represents the remaining ciliary neurotrophic factor analogue activity amount compared with the initial one.

Table 1 stabilizer Quantity (by weight ratio) active(%) Stored at 4°C for 3 months Stored at 25°C for 1 month Mannitol 500 89 85 Sorbitol 500 88 83 Glycine 200 89 83 cysteine 200 81 74 polyoxyethylene sorbitan fatty acid ester 0.4 85 78

Glycerin 2000 95 92 Heparin 1000 82 77 Sodium alginate 200 89 84 Hyaluronic Acid 200 78 71 PEG6000 500 90 86 human serum albumin 500 86 78 Polyvinylpyrrolidone (50,000) 500 83 79 Sodium carboxymethyl cellulose 200 82 77 Glutathione (reduced form) 200 78 73 Human Serum Albumin NaCl 50090 87 77 human serum albumin cysteine 500100 90 81 Mannitol Ascorbic Acid 50010 94 87 Mannitol Polyoxyethylene Sorbitan Fatty Acid Ester 5000.4 98 96 Mannitol polyoxyethylene sorbitan fatty acid ester ascorbic acid 5000.410 100 97 do not add - 79 68

Example 2

Several stabilizing agents listed in table 2 are added to 0.2 mg/ml ciliary neurotrophic factor analog (its amino acid composition is the same as sequence 2 in the accompanying drawing), the mixture contains the inorganic salt buffer solution of 10mM different pH, without After bacterial filtration, it is sealed in a bottle at a rate of 0.5 ml/cartridge. Observe its changes under different conditions. Measured by method (B), the results are shown in Table 2.

Table 2 stabilizer Quantity (by weight ratio) Solution pH active(%) Stored at 4°C for 3 months Stored at 25°C for 1 month Mannitol polyoxyethylene sorbitan fatty acid ester ascorbyl acetate buffer 5000.41010Mm 4.0 65 42 Mannitol polyoxyethylene sorbitan fatty acid ester ascorbyl acetate buffer 5000.41010Mm 5.0 83 78 Mannitol Polyoxyethylene Sorbitan Fatty Acid Ester Ascorbyl Phosphate Buffer 5000.41010Mm 6.0 95 92 Mannitol Polyoxyethylene Sorbitan Fatty Acid Ester Ascorbyl Phosphate Buffer 5000.41010Mm 7.0 100 97 Mannitol Polyoxyethylene Sorbitan Fatty Acid Ester Ascorbyl Phosphate Buffer 5000.41010Mm 8.0 98 96 Mannitol polyoxyethylene sorbitan fatty acid ester ascorbyl glycine-NaOH buffer 5000.41010Mm 9.0 93 90

Example 3

The various stabilizers listed in Table 3 are added to 0.2 mg/ml ciliary neurotrophic factor analog (its amino acid composition is the same as sequence 1 in the accompanying drawing), and the mixture contains pH7.0 in 20mM phosphate buffer , after sterile filtration, it is divided into glass vials at 0.5 ml/branch. Then lyophilize according to the following procedure: freezing at -35°C or below -35°C for 4 hours, followed by the first drying for 10 hours, heating from -35°C to -5°C, increasing the pressure from 0.015 Torr to 0.2 Torr, and then For the second drying, the temperature is heated from -5°C to 30°C, the pressure is reduced from 0.2 torr to 0.05 torr, and then sterilized dry nitrogen is filled into the inside of the vial to make it reach an atmospheric pressure, and the vial is placed under aseptic conditions. Stoppered and finally sealed with an aluminum cap. Observe its changes under different conditions. Measured by method (B), the results are shown in Table 3.

table 3 stabilizer Quantity (by weight ratio) active(%) Stored at 4°C for 6 months Stored at 37°C for 1 month Mannitol 500(5%) 91 86 Sorbitol 500 92 86 Glycine 200 92 87 cysteine 200 86 74 sodium alginate 200 91 86 human serum albumin 500 95 91 Glutathione (reduced form) 200 88 84 human serum albumin NaCl 50090 95 92 human serum albumin cysteine 500100 96 93 Mannitol Ascorbic Acid 50010 96 93 Mannitol Polyoxyethylene Sorbitan Fatty Acid Ester 5000.4 97 96 Mannitol polyoxyethylene sorbitan fatty acid ester ascorbic acid 5000.410 99 97 do not add - 82 76

Sequence Listing

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Claims (5)

1. one kind contains ciliary neurotrophic factor (Ciliary Neurotrophic Factor, CNTF) stable pharmaceutical of analog, it is characterized in that it except ciliary neurotrophic factor (CNTF) analog as the effective ingredient, also comprise the acceptable fatty acid esters of sorbitan of medicine, mannitol, L-ascorbic acid and phosphate, its pH value of solution or reconstruction back pH value of solution are between 5-8, and the ciliary neurotrophic factor analog is meant the albumen with 1-3 arbitrary sequence structure in the sequence table.

2. a kind of stable pharmaceutical that contains the ciliary neurotrophic factor analog according to claim 1, it is characterized in that said fatty acid esters of sorbitan addition by weight and the ratio of ciliary neurotrophic factor analog between 0.01-1000.

3. a kind of stable pharmaceutical that contains the ciliary neurotrophic factor analog according to claim 1, it is characterized in that said mannitol addition by weight and the ratio of ciliary neurotrophic factor analog between 10-5000.

4. a kind of stable pharmaceutical that contains the ciliary neurotrophic factor analog according to claim 1, it is characterized in that said L one ascorbic acid addition by weight and the ratio of ciliary neurotrophic factor analog between 0.001-100.

5. a kind of stable pharmaceutical that contains the ciliary neurotrophic factor analog according to claim 1, it is characterized in that said phosphate adding quantity by weight and the ratio of ciliary neurotrophic factor analog between 0.01-1000.