Thymalfasin composition, preparation as well as preparation method and application of thymalfasin composition

Abstract

The invention discloses a thymalfasin composition, a preparation as well as a preparation method and application of the thymalfasin composition. The thymalfasin composition comprises thymalfasin, trehalose and mannitol. According to the thymalfasin freeze-dried powder injection disclosed by the invention, trehalose and mannitol are used as auxiliary materials of thymalfasin, so that the content of polymers in a thymalfasin preparation is relatively low, and no significant change occurs in a long-term stability experiment, so that the stability of the thymalfasin freeze-dried powder injection is improved; according to the invention, the buffer solution is used as a solution preparation solvent, so that the pH value of the whole solution preparation system is always kept at 6.0-7.5, and the stability of thymalfasin in the preparation process is improved.

Description

Technical Field

[0001] The invention relates to a thymosin composition, a preparation, a preparation method and an application thereof. Background Art

[0002] Thymosin alpha 1 is a synthetic 28-peptide compound and one of the most biologically active polypeptides in the fifth component of thymosin. Its main function is to promote T cell maturation and enable cells to produce various lymphokines, such as alpha and gamma interferons and interleukin 2 (IL-2), IL-3, etc. after being activated by various antigens or mitogens. It can improve the body's immunity and enhance the body's resistance to various foreign pathogens. It is clinically used alone to treat chronic hepatitis B, or in combination with antiviral drugs and anti-cancer drugs, it has a good effect.

[0003] Thymosin alpha-1 was first developed and produced by Sciclone Pharmaceuticals in the United States. The specification is 1.6mg sterile powder for injection, and the trade name is "Ridaxin". It entered the Chinese market at the end of the last century. At present, many domestic pharmaceutical companies have obtained the production approval for thymosin alpha-1 for injection.

[0004] Thymosin is unstable in aqueous solution, so preparing it into a lyophilized preparation solves this problem to a certain extent. However, the stability of the thymosin solution during the preparation process is also key. Studies have found that thymosin has good stability in low temperature, neutral and weakly acidic environments. Therefore, during the preparation process, not only stabilizers should be added to protect the stability of the drug solution, but it is also very important to control the pH of the solution preparation process.

[0005] Chinese patent 201110427640.7 provides a thymosin α1 injection preparation pharmaceutical composition, specifically: after dissolving excipients, antioxidants, and local analgesics in water, adding buffer, adjusting the pH to 6.5 with acetic acid, constant volume, adding activated carbon to remove pyrogens, filtering, filling, and freeze drying to obtain thymosin α1 freeze-dried preparation. The patent has complex components and cumbersome operations, and is not suitable for large-scale production.

[0006] Chinese patent 201110220652.2 provides a pharmaceutical composition containing thymosin, specifically: thymosin, mannitol and sorbitol are dissolved in water, adjusted to 3.8-4.2 with 0.01M acetic acid-sodium acetate buffer solution, filled into a controlled injection bottle, fully plugged, rolled to obtain an injection or half plugged and freeze-dried to obtain a lyophilized powder injection. Although the injection and lyophilized powder obtained by the invention have stable quality, their acidic drug properties are not suitable for clinical use.

[0007] Chinese patent 201210499830.4 provides a method for preparing a thymosin freeze-dried preparation: tert-butyl alcohol, mannitol, and sodium acetate are added to 50% water for injection in sequence, stirred and dissolved, and then thymosin is added and stirred and dissolved, and then the pH of the solution is adjusted to 5.5-6.5 with acetic acid or sodium acetate, and finally the volume is fixed to 1000 ml. The added tert-butyl alcohol improves the stability of the main drug. The patent uses tert-butyl alcohol to improve the stability of the drug, but ignores the production safety of tert-butyl alcohol as an organic solvent.

[0008] Chinese patent 202011063337.9 provides a lyophilized composition containing thymosin, which is composed of an active ingredient thymosin and a pH regulator, preferably meglumine, to adjust the pH to 7.5-7.7, preferably 7.6. This patent has few excipients and only demonstrates superiority in terms of reconstitution time, visible foreign matter and insoluble particles. Summary of the invention

[0009] The present invention aims to solve the problem of poor stability of existing thymosin, and further provides a thymosin composition, preparation and preparation method and application thereof. The thymosin preparation of the present invention has good stability, simple ingredients and simple preparation process.

[0010] The present invention solves the above technical problems through the following technical solutions:

[0011] The invention discloses a thymosin composition, which comprises thymosin, trehalose and mannitol. The pH value of the thymosin composition is 6.0-7.5.

[0012] In the present invention, the chemical name of thymosin is N-acetyl-L-seryl-L-α-aspartyl-L-alanyl-L-alanyl-L-valyl-L-α-aspartyl-L-threonyl-L-seryl-L-seryl-L-α-glutamyl-L-isoleucyl-L-threonyl-L-threonyl-L-lysyl-L-α-aspartyl-L-leucyl-L-lysyl-L-α-glutamyl-L-lysyl-L-α-lysyl-L-lysyl-L-α-glutamyl-L-lysyl-L-α-glutamyl-L-valyl-L-valyl-L-α-glutamyl-L-α-glutamyl-L-alanyl-L-α-glutamyl-L-asparagine, which is a polypeptide compound composed of 28 amino acids, and the chemical formula is C 129 H 215 N 33 O 55 , CAS No.69440-99-9.

[0013] In the present invention, the mass ratio of the thymosin to the trehalose is preferably 1.6:(15-100), such as 1.6:15, 1.6:20, 1.6:25, 1.6:30, 1.6:35 or 1.6:40.

[0014] In the present invention, the mass ratio of the thymosin to the mannitol is preferably 1.6:(15-100), such as 1.6:15, 1.6:20, 1.6:25, 1.6:30, 1.6:35 or 1.6:40.

[0015] In the present invention, the mass ratio of the thymosin, the trehalose and the mannitol is preferably 1.6:(15-100):(15-100), for example, 1.6:15:35, 1.6:25:25, 1.6:35:15, 1.6:20:40, 1.6:30:30 or 1.6:40:20.

[0016] In the present invention, the mannitol is preferably 20% mannitol injection, % refers to the ratio of weight to volume, the unit is kg / L.

[0017] In the present invention, the pH value of the thymosin composition is 6.7, 7.0 or 7.3;

[0018] In the present invention, the thymosin alpha-sin composition preferably further comprises a buffer, and the buffer is preferably two or more of citric acid, citrate buffer, phosphate buffer and sodium hydroxide. Among them, the citrate buffer is preferably sodium citrate. The phosphate buffer is preferably one or more of sodium dihydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate and dipotassium hydrogen phosphate, preferably sodium dihydrogen phosphate and disodium hydrogen phosphate buffer.

[0019] Wherein, the buffer is preferably a mixture of sodium dihydrogen phosphate solution and sodium hydrogen phosphate solution.

[0020] The pH of the buffer is preferably 6.0-7.5, such as 6.7, 7.0 or 7.3.

[0021] The amount of the buffer added is determined according to the concentration requirement of the active ingredient thymosin.

[0022] In the present invention, the concentration of thymalfasin is preferably 1.0-2.0 g / L, for example 1.6 g / L.

[0023] In the present invention, the concentration of trehalose is preferably 15-40 g / L, such as 15 g / L, 20 g / L, 25 g / L, 30 g / L, 35 g / L or 40 g / L.

[0024] In the present invention, the concentration of mannitol is preferably 15-40 g / L, for example 15 g / L, 20 g / L, 25 g / L, 30 g / L, 35 g / L or 40 g / L.

[0025] The present invention also provides a method for preparing the thymosin composition, which comprises: mixing the thymosin, the trehalose, the mannitol and the buffer to obtain the thymosin composition.

[0026] In the present invention, the mixing method preferably comprises: mixing and dissolving the trehalose, the mannitol and a part of the buffer, adding the thymosin to dissolve, and then adding the remaining part of the buffer to obtain a mixed solution.

[0027] The invention also discloses a thymosin preparation, which comprises: thymosin, trehalose, mannitol and a buffer; the thymosin, trehalose, mannitol and a buffer are the aforementioned thymosin, trehalose, mannitol and a buffer.

[0028] In the present invention, the pH of the thymalfasin preparation is preferably 6.0 to 7.5, such as 6.7, 7.0 or 7.3.

[0029] In the present invention, the concentration of thymalfasin is preferably 1.0-2.0 g / L, for example 1.6 g / L.

[0030] In the present invention, the concentration of trehalose is preferably 15-40 g / L, such as 15 g / L, 20 g / L, 25 g / L, 30 g / L, 35 g / L or 40 g / L.

[0031] In the present invention, the concentration of mannitol is preferably 15-40 g / L, for example 15 g / L, 20 g / L, 25 g / L, 30 g / L, 35 g / L or 40 g / L.

[0032] In the present invention, the thymosin preparation is preferably composed of the following components: thymosin, trehalose, mannitol and a buffer, wherein the concentration of thymosin is 1.6 g / L, the concentration of trehalose is 15 g / L, the concentration of mannitol is 35 g / L, and the pH of the buffer is 6.7.

[0033] In the present invention, the thymosin preparation is preferably composed of the following components: thymosin, trehalose, mannitol and a buffer, the concentration of the thymosin is 1.6 g / L, the concentration of the trehalose is 25 g / L, the concentration of the mannitol is 25 g / L, and the pH of the buffer is 6.7.

[0034] In the present invention, the thymosin preparation is preferably composed of the following components: thymosin, trehalose, mannitol and a buffer, the concentration of the thymosin is 1.6 g / L, the concentration of the trehalose is 35 g / L, the concentration of the mannitol is 15 g / L, and the pH of the buffer is 7.0.

[0035] In the present invention, the thymosin preparation is preferably composed of the following components: thymosin, trehalose, mannitol and a buffer, wherein the concentration of thymosin is 1.6 g / L, the concentration of trehalose is 20 g / L, the concentration of mannitol is 40 g / L, and the pH of the buffer is 7.0.

[0036] In the present invention, the thymosin preparation is preferably composed of the following components: thymosin, trehalose, mannitol and a buffer, the concentration of the thymosin is 1.6 g / L, the concentration of the trehalose is 30 g / L, the concentration of the mannitol is 30 g / L, and the pH of the buffer is 7.3.

[0037] In the present invention, the thymosin preparation is preferably composed of the following components: thymosin, trehalose, mannitol and a buffer, the concentration of the thymosin is 1.6 g / L, the concentration of the trehalose is 40 g / L, the concentration of the mannitol is 20 g / L, and the pH of the buffer is 7.3.

[0038] The present invention also discloses a method for preparing the thymus method preparation, which comprises: mixing the thymus method preparation, the trehalose method, the mannitol method and the buffering agent to obtain a mixed solution, and freeze-drying the mixed solution to obtain the thymus method preparation.

[0039] In the present invention, the method for preparing the mixed solution preferably comprises: mixing and dissolving the trehalose, the mannitol and part of the buffer, adding the thymosin to dissolve, and then adding the remaining part of the buffer to obtain a mixed solution.

[0040] In the present invention, the temperature during the preparation of the thymus preparation is preferably controlled at 10-30°C, more preferably 15-25°C.

[0041] In the present invention, the freeze-drying process preferably includes pre-freezing, sublimation drying and analytical drying performed in sequence.

[0042] The pre-freezing temperature is preferably below -45°C. The pre-freezing temperature is preferably 3 to 6 hours.

[0043] The temperature of the sublimation drying is preferably -12 to 20° C., such as -15° C. The time of the sublimation drying is preferably 10 to 18 hours.

[0044] The drying temperature is preferably 20-30° C., such as 25° C. The drying time is preferably 8-15 h.

[0045] In the present invention, filtering and / or filling treatment is preferably performed before freeze-drying.

[0046] Wherein, the filtration is preferably performed using a 0.2 μm filter membrane.

[0047] Wherein, a semi-plugging treatment can be performed after the filling.

[0048] The present invention also discloses the use of the thymosin composition or the thymosin preparation in preparing medicine for treating chronic hepatitis B.

[0049] The present invention also discloses a combined pharmaceutical composition, which comprises: an antiviral drug and the aforementioned thymosin alpha-sin composition;

[0050] Alternatively, antiviral drugs and the aforementioned thymalfasin preparations;

[0051] Or, a combination of an anti-tumor drug and the aforementioned thymofaxin;

[0052] Alternatively, an antineoplastic drug and the aforementioned thymosin preparation.

[0053] In the present invention, the aforementioned combined pharmaceutical composition is used to prepare antiviral drugs or anti-tumor drugs.

[0054] The positive and progressive effects of the present invention are:

[0055] (1) The present invention uses trehalose and mannitol as excipients of thymosin, so that the polymer content in the thymosin preparation is low and no significant change occurs in the long-term stability experiment, thereby improving the stability of the thymosin freeze-dried powder injection.

[0056] (2) The present invention uses a buffer solution as the preparation solvent, so that the pH of the entire preparation system is always maintained at 6.0-7.5, thereby improving the stability of thymosin during the preparation process.

[0057] (3) The pharmaceutical composition of the present invention has simple ingredients, simple process operation, low cost, and is suitable for large-scale production. DETAILED DESCRIPTION

[0058] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0059] Example 1

[0060] The composition of the thymus fasin composition of the present embodiment is as follows:

[0061]

[0062] Preparation method:

[0063] (1) Prepare 1000 ml of phosphate buffer with a concentration of 0.02 mol / L and a pH of 6.7 using sodium dihydrogen phosphate and disodium hydrogen phosphate as raw materials and set aside;

[0064] (2) Weigh 15 g of excipient trehalose and 175 ml of 20% mannitol injection;

[0065] (3) Take about 800 ml of phosphate buffer with a pH of 6.7, add the auxiliary materials in step (2), and stir to completely dissolve them;

[0066] (4) Weigh 1.6 g of thymosin and add it to the solution in step (3), stirring until it is completely dissolved;

[0067] (5) Dilute the above solution to 1000 ml with phosphate buffer at pH 6.7;

[0068] The temperature of the preparation process of the above steps (1) to (5) is controlled at 15-25°C;

[0069] (6) Sterile filtration with a 0.2 μm filter membrane, filling into 2 ml borosilicate glass tube injection bottles (the content of thymosin is 1.6 g / L, and the pH of the solution is 6.7), half-stoppering, freeze-drying, filling with nitrogen, fully stoppering, unpacking, and capping to obtain thymosin lyophilized powder injection.

[0070] The freeze-drying process is as follows: pre-freeze the plate layer to below -45°C, keep it for 3 to 6 hours to completely freeze the sample, turn on the condenser, wait for the condenser to drop below -50°C, turn on the vacuum pump, and let the vacuum reach below 20Pa. Heat up to -15°C for sublimation drying, keep it warm for 10 to 18 hours, heat up to 25°C, perform analytical drying, and keep it warm for 8 to 15 hours.

[0071] Example 2

[0072] The composition of the thymus fasin composition of the present embodiment is as follows:

[0073]

[0074] Preparation method:

[0075] (1) Prepare 1000 ml of phosphate buffer with a concentration of 0.02 mol / L and a pH of 6.7 using sodium dihydrogen phosphate and disodium hydrogen phosphate as raw materials and set aside;

[0076] (2) Weigh 25 g of excipient trehalose and 125 ml of 20% mannitol injection;

[0077] (3) Take about 800 ml of phosphate buffer with a pH of 6.7, add the auxiliary materials in step (2), and stir to completely dissolve them;

[0078] (4) Weigh 1.6 g of thymosin and add it to the solution in step (3), stirring until it is completely dissolved;

[0079] (5) Dilute the above solution to 1000 ml with phosphate buffer at pH 6.7;

[0080] (6) Sterilize and filter with a 0.2 μm filter membrane, fill into a 2 ml borosilicate glass tube injection bottle (the content of thymosin is 1.6 g / L, and the pH of the solution is 6.7), half-stopper, freeze-dry, fill with nitrogen, fully stopper, unpack, and cap to obtain thymosin freeze-dried powder injection; wherein the freeze-drying process is the same as that in Example 1.

[0081] Example 3

[0082] The composition of the thymus fasin composition of the present embodiment is as follows:

[0083]

[0084] Preparation method:

[0085] (1) Prepare 1000 ml of phosphate buffer with a concentration of 0.02 mol / L and a pH of 7.0 using sodium dihydrogen phosphate and disodium hydrogen phosphate as raw materials and set aside;

[0086] (2) Weigh 35 g of excipient trehalose and 75 ml of 20% mannitol injection;

[0087] (3) Take about 800 ml of phosphate buffer with a pH of 7.0, add the auxiliary materials in step (2), and stir to completely dissolve them;

[0088] (4) Weigh 1.6 g of thymosin and add it to the solution in step (3), stirring until it is completely dissolved;

[0089] (5) Dilute the above solution to 1000 ml with phosphate buffer (pH 7.0);

[0090] (6) Filter the mixture through a 0.2 μm filter membrane for sterilization, fill the mixture into a 2 ml borosilicate glass tube injection bottle (the content of thymosin is 1.6 g / L, and the pH value of the solution is 7.0), half-stopper the solution, freeze-dry, fill with nitrogen, fully stopper the solution, unpack the solution, and cap the solution to obtain thymosin freeze-dried powder for injection; wherein the freeze-drying process is the same as that in Example 1.

[0091] Example 4

[0092] The composition of the thymus fasin composition of the present embodiment is as follows:

[0093]

[0094] Preparation method:

[0095] (1) Prepare 1000 ml of phosphate buffer with a concentration of 0.02 mol / L and a pH of 7.0 using sodium dihydrogen phosphate and disodium hydrogen phosphate as raw materials and set aside;

[0096] (2) Weigh 20 g of excipient trehalose and 200 ml of 20% mannitol injection;

[0097] (3) Take about 700 ml of phosphate buffer with a pH of 7.0, add the auxiliary materials in step (2), and stir to completely dissolve them;

[0098] (4) Weigh 1.6 g of thymosin and add it to the solution in step (3), stirring until it is completely dissolved;

[0099] (5) Dilute the above solution to 1000 ml with phosphate buffer (pH 7.0);

[0100] (6) Filter the mixture through a 0.2 μm filter membrane for sterilization, fill the mixture into a 2 ml borosilicate glass tube injection bottle (the content of thymosin is 1.6 g / L, and the pH value of the solution is 7.0), half-stopper the solution, freeze-dry, fill with nitrogen, fully stopper the solution, unpack the solution, and cap the solution to obtain thymosin freeze-dried powder for injection; wherein the freeze-drying process is the same as that in Example 1.

[0101] Example 5

[0102] The composition of the thymus fasin composition of the present embodiment is as follows:

[0103]

[0104] Preparation method:

[0105] (1) Prepare 1000 ml of phosphate buffer with a concentration of 0.02 mol / L and a pH of 7.3 using sodium dihydrogen phosphate and disodium hydrogen phosphate as raw materials and set aside;

[0106] (2) Weigh 30 g of excipient trehalose and 150 ml of 20% mannitol injection;

[0107] (3) Take about 800 ml of phosphate buffer with a pH of 7.3, add the auxiliary materials in step (2), and stir to completely dissolve them;

[0108] (4) Weigh 1.6 g of thymosin and add it to the solution in step (3), stirring until it is completely dissolved;

[0109] (5) Dilute the above solution to 1000 ml with phosphate buffer at pH 7.3;

[0110] (6) Sterilize and filter with a 0.2 μm filter membrane, fill into a 2 ml borosilicate glass tube injection bottle (the content of thymosin is 1.6 g / L, and the pH of the solution is 7.3), half-stopper, freeze-dry, fill with nitrogen, fully stopper, unpack, and cap to obtain thymosin freeze-dried powder injection; wherein the freeze-drying process is the same as that in Example 1.

[0111] Example 6

[0112] The composition of the thymus fasin composition of the present embodiment is as follows:

[0113]

[0114] Preparation method:

[0115] (1) Prepare 1000 ml of phosphate buffer with a concentration of 0.02 mol / L and a pH of 7.3 using sodium dihydrogen phosphate and disodium hydrogen phosphate as raw materials and set aside;

[0116] (2) Weigh excipients: 40 g of trehalose and 100 ml of 20% mannitol injection;

[0117] (3) Take about 800 ml of phosphate buffer with a pH of 7.3, add the auxiliary materials in step (2), and stir to completely dissolve them;

[0118] (4) Weigh 1.6 g of thymosin and add it to the solution in step (3), stirring until it is completely dissolved;

[0119] (5) Dilute the above solution to 1000 ml with phosphate buffer at pH 7.3;

[0120] (6) Filter the mixture through a 0.2 μm filter membrane for sterilization, fill the mixture into a 2 ml borosilicate glass tube injection bottle (the content of thymosin is 1.6 g / L, and the pH value of the solution is 7.3), half-stopper the solution, freeze-dry, fill with nitrogen, fully stopper the solution, unpack the solution, and cap the solution to obtain thymosin freeze-dried powder for injection; wherein the freeze-drying process is the same as that in Example 1.

[0121] Comparative Example 1

[0122] The thymus fasin composition of this comparative example is composed as follows:

[0123]

[0124] Preparation method:

[0125] (1) Prepare 1000 ml of phosphate buffer solution with a concentration of 0.1 mol / L and a pH of 7.0 using sodium dihydrogen phosphate and disodium hydrogen phosphate as raw materials and set aside;

[0126] (2) Take 200 ml of 20% mannitol injection and dilute it to 700 ml with water for injection;

[0127] (3) Add 200 ml of phosphate buffer (pH = 7.0) to the above solution and stir evenly;

[0128] (4) Weigh 1.6 g of thymosin and add it to the above solution, stirring until it is completely dissolved;

[0129] (5) Dilute the above solution to 1000 ml with water for injection;

[0130] (6) Filter the mixture through a 0.2 μm filter membrane for sterilization, fill the mixture into a 2 ml borosilicate glass tube injection bottle, half-stopper the bottle, lyophilize, and cap the bottle to obtain thymosin lyophilized powder injection; wherein the lyophilization process is the same as that in Example 1.

[0131] Comparative Example 2

[0132] The thymus fasin composition of this comparative example is composed as follows:

[0133]

[0134] Preparation method:

[0135] (1) Prepare 1000 ml of phosphate buffer with a concentration of 0.02 mol / L and a pH of 7.0 using sodium dihydrogen phosphate and disodium hydrogen phosphate as raw materials and set aside;

[0136] (2) Weigh 20 g of sodium chloride as an excipient and 200 ml of 20% mannitol injection;

[0137] (3) Take about 700 ml of phosphate buffer with a pH of 7.0, add the auxiliary materials in step (2), and stir to completely dissolve them;

[0138] (4) Weigh 1.6 g of thymosin and add it to the solution in step (3), stirring until it is completely dissolved;

[0139] (5) Dilute the above solution to 1000 ml with phosphate buffer (pH 7.0);

[0140] (6) Filter the mixture through a 0.2 μm filter membrane for sterilization, fill the mixture into a 2 ml borosilicate glass tube injection bottle (the content of thymosin is 1.6 g / L, and the pH value of the solution is 7.0), half-stopper the solution, freeze-dry, fill with nitrogen, fully stopper the solution, unpack the solution, and cap the solution to obtain thymosin freeze-dried powder for injection; wherein the freeze-drying process is the same as that in Example 1.

[0141] Comparative Example 3

[0142] The thymus fasin composition of this comparative example is composed as follows:

[0143]

[0144] Preparation method:

[0145] (1) Prepare 1000 ml of phosphate buffer with a concentration of 0.02 mol / L and a pH of 7.0 using sodium dihydrogen phosphate and disodium hydrogen phosphate as raw materials and set aside;

[0146] (2) Weigh 20 g of trehalose and 40 g of sorbitol as auxiliary materials;

[0147] (3) Take about 800 ml of phosphate buffer with a pH of 7.0, add the auxiliary materials in step (2), and stir to completely dissolve them;

[0148] (4) Weigh 1.6 g of thymosin and add it to the solution in step (3), stirring until it is completely dissolved;

[0149] (5) Dilute the above solution to 1000 ml with phosphate buffer (pH 7.0);

[0150] (6) Filter the mixture through a 0.2 μm filter membrane for sterilization, fill the mixture into a 2 ml borosilicate glass tube injection bottle (the content of thymosin is 1.6 g / L, and the pH value of the solution is 7.0), half-stopper the solution, freeze-dry, fill with nitrogen, fully stopper the solution, unpack the solution, and cap the solution to obtain thymosin freeze-dried powder for injection; wherein the freeze-drying process is the same as that in Example 1.

[0151] Effect Example 1

[0152] The thymus alpha-sin freeze-dried powder obtained in the above example was tested for key items, and the methods and limits were as follows:

[0153] (1) Properties: It should be white or off-white loose lumps.

[0154] (2) Acidity: The pH value should be 6.0-7.5.

[0155] (3) Solution clarity and color: The solution should be clear and colorless. If turbid, it should not be more concentrated than turbidity standard solution No. 1.

[0156] (4) Moisture content: should not exceed 5.0%.

[0157] (5) Related substances: Determined by high performance liquid chromatography, using octadecylsilane bonded silica gel as filler; ammonium sulfate buffer-acetonitrile (90:10) as mobile phase A, ammonium sulfate buffer-acetonitrile (50:50) as mobile phase B, gradient elution; column temperature 50°C; detection wavelength 210nm, injection volume 20μl. Single impurity shall not exceed 2.0%, total impurity shall not exceed 4.0%.

[0158] (6) Polymer: Determined by molecular exclusion method, using spherical hydrophilic modified silica gel as filler, trifluoroacetic acid-acetonitrile-water as mobile phase, detection wavelength at 210nm, injection volume at 20μl. The polymer content shall not exceed 0.5%.

[0159] (7) Content: Determined by high performance liquid chromatography. The chromatographic conditions are shown under related substances. The content should be 90.0%-110.0% of the labeled amount.

[0160] The thymosin freeze-dried preparations obtained in Examples 1 to 6 and Comparative Examples 1-3 were subjected to an accelerated 6-month stability test under the conditions of 25°C ± 2°C and RH 60% ± 5%. The test results are as follows:

[0161]

[0162]

[0163] The results in Table 1 show that the results of the accelerated 6-month test of the samples prepared in Examples 1-6 are all within the range of the pharmacopoeia standard (single impurity shall not exceed 2.0%, total impurity shall not exceed 4.0%, and the content shall be 90.0%-110.0%), and no significant changes have occurred. Compared with the comparative example, the related substances, polymers and content detection items are all better than the comparative example.

[0164] The thymosin freeze-dried preparations obtained in Examples 1 to 6 and Comparative Examples 1 to 3 were subjected to a long-term 24-month stability test. The long-term test conditions were 5°C ± 3°C. The test results were as follows:

[0165]

[0166]

[0167]

[0168] The results in Table 2 show that the long-term 24-month test results of the samples prepared under the conditions of Examples 1-6 are all within the pharmacopoeia standard range and have not changed significantly. Compared with the comparative examples, the relevant substances, polymers and content detection items are all better than the comparative examples.

[0169] Peptide drugs contain disulfide bonds, exposed amino groups and carboxyl groups, and are prone to form covalent polymers due to intermolecular disulfide bonds or dehydration between amino and carboxyl groups. Polymer impurities formed by covalent bonds may have a greater immunogenicity risk and should be given special attention in the quality research of peptide drug preparations. In the above examples, special attention was paid to polymers. In the stability study of the samples in each example, no significant changes occurred in the polymers, and the quality was stable.

Claims

1. A thymic fasin composition, characterized in that: It includes: Thymosin, trehalose and mannitol, wherein the pH value of the thymosin composition is 6.0-7.

5.

2. The thymic fasin composition according to claim 1, characterized in that The thymosin composition satisfies one or more of ① to ⑥: ① The mass ratio of the thymosin to the trehalose is 1.6:(15-100), for example, 1.6:15, 1.6:20, 1.6:25, 1.6:30, 1.6:35 or 1.6:40; ② The mass ratio of the thymosin to the mannitol is 1.6:(15-100), for example, 1.6:15, 1.6:20, 1.6:25, 1.6:30, 1.6:35 or 1.6:40; ③ The mass ratio of the thymosin, the trehalose and the mannitol is 1.6:(15-100):(15-100), for example, 1.6:15:35, 1.6:25:25, 1.6:35:15, 1.6:20:40, 1.6:30:30 or 1.6:40:20; ④ The mannitol is 20% mannitol injection, % refers to the ratio of weight to volume, the unit is kg / L; ⑤ The pH value of the thymus fasin composition is 6.7, 7.0 or 7.3; ⑥ The thymus alpha-sin composition also includes a buffer, which is preferably two or more of citric acid, citrate buffer, phosphate buffer and sodium hydroxide, the citrate buffer is preferably sodium citrate, the phosphate buffer is preferably one or more of sodium dihydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, and dipotassium hydrogen phosphate, and is more preferably a buffer formed by sodium dihydrogen phosphate and disodium hydrogen phosphate; the pH of the buffer is preferably 6.0 to 7.5, for example 6.7, 7.0 or 7.

3.

3. A method for preparing the thymic alpha-sin composition according to claim 1 or 2, characterized in that: It includes: Mixing the thymosin, the trehalose, the mannitol and the buffer to obtain the thymosin composition; The mixing method preferably comprises: mixing and dissolving the trehalose, the mannitol and a part of the buffer, adding the thymosin to dissolve, and then adding the remaining part of the buffer to obtain a mixed solution.

4. A thymosin preparation, characterized in that: It includes: thymalfasin, trehalose, and mannitol; The thymosin, trehalose and mannitol are the thymosin, trehalose and mannitol according to claim 1 or 2.

5. The thymosin preparation according to claim 4, characterized in that The thymosin preparation meets one or more of ① to ⑤: ① The pH of the thymomethoxazole preparation is 6.0 to 7.5, for example, 6.7, 7.0 or 7.3; ② The concentration of thymosin is 1.0-2.0 g / L, for example 1.6 g / L; ③ The concentration of trehalose is 15-40 g / L, for example, 15 g / L, 20 g / L, 25 g / L, 30 g / L, 35 g / L or 40 g / L; ④ the concentration of mannitol is 15-40 g / L, for example 15 g / L, 20 g / L, 25 g / L, 30 g / L, 35 g / L or 40 g / L; ⑤ The thymus alpha-sin preparation also includes a buffer, and the buffer is preferably two or more of citric acid, citrate buffer, phosphate buffer and sodium hydroxide, the citrate buffer is preferably sodium citrate, and the phosphate buffer is preferably one or more of sodium dihydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, and dipotassium hydrogen phosphate, and more preferably a buffer formed by sodium dihydrogen phosphate and disodium hydrogen phosphate; the pH of the buffer is preferably 6.0 to 7.5, for example 6.7, 7.0 or 7.

3.

6. The thymosin preparation according to claim 4, characterized in that The thymosin preparation is composed of the following components: thymosin, trehalose, mannitol and a buffer, wherein the concentration of the thymosin is 1.6 g / L, the concentration of the trehalose is 15 g / L, the concentration of the mannitol is 35 g / L, and the pH of the buffer is 6.7; Or, the thymosin preparation is composed of the following components: thymosin, trehalose, mannitol and a buffer, wherein the concentration of the thymosin is 1.6 g / L, the concentration of the trehalose is 25 g / L, the concentration of the mannitol is 25 g / L, and the pH of the buffer is 6.7; Or, the thymosin preparation is composed of the following components: thymosin, trehalose, mannitol and a buffer, wherein the concentration of the thymosin is 1.6 g / L, the concentration of the trehalose is 35 g / L, the concentration of the mannitol is 15 g / L, and the pH of the buffer is 7.0; Or, the thymosin preparation is composed of the following components: thymosin, trehalose, mannitol and a buffer, wherein the concentration of the thymosin is 1.6 g / L, the concentration of the trehalose is 20 g / L, the concentration of the mannitol is 40 g / L, and the pH of the buffer is 7.0; Or, the thymosin preparation is composed of the following components: thymosin, trehalose, mannitol and a buffer, wherein the concentration of the thymosin is 1.6 g / L, the concentration of the trehalose is 30 g / L, the concentration of the mannitol is 30 g / L, and the pH of the buffer is 7.3; Or, the thymosin preparation consists of the following components: thymosin, trehalose, mannitol and a buffer, the concentration of the thymosin is 1.6 g / L, the concentration of the trehalose is 40 g / L, the concentration of the mannitol is 20 g / L, and the pH of the buffer is 7.

3.

7. A method for preparing a thymus preparation according to any one of claims 4 to 6, characterized in that: It includes: The thymosin, the trehalose, the mannitol and the buffer are mixed to obtain a mixed solution, which is freeze-dried to obtain the thymosin preparation.

8. The method for preparing the thymus gland preparation according to claim 7, characterized in that: The preparation method of the thymus preparation satisfies one or more of ① to ④: ① The preparation method of the mixed solution comprises: mixing and dissolving the trehalose, the mannitol and part of the buffer, adding the thymosin to dissolve, and then adding the remaining part of the buffer to obtain a mixed solution; ② During the preparation of the thymus preparation, the temperature is controlled at 10-30°C, preferably 15-25°C; ③ The freeze-drying process includes pre-freezing, sublimation drying and desorption drying in sequence, wherein the pre-freezing temperature is preferably below -45°C, the pre-freezing temperature is preferably 3-6h, the sublimation drying temperature is preferably -12-20°C, for example -15°C, the sublimation drying time is preferably 10-18h, the desorption drying temperature is preferably 20-30°C, for example 25°C, and the desorption drying time is preferably 8-15h; ④ Before freeze-drying, filtering and / or filling are performed, and the filtration is preferably performed using a 0.2 μm filter membrane.

9. Use of the thymosin composition according to any one of claims 1 to 3 or the thymosin preparation according to any one of claims 4 to 6 in the preparation of a medicament for treating chronic hepatitis B.

10. A combined pharmaceutical composition, characterized in that: It includes: An antiviral drug and a thymosin composition according to any one of claims 1 to 3; Or, an antiviral drug and the thymosin preparation according to any one of claims 4 to 6; Or, an anti-tumor drug and a thymosin composition according to any one of claims 1 to 3; Alternatively, an antitumor drug and the thymosin preparation according to any one of claims 4 to 6.

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