A levothyroxine sodium for injection sustained-release microsphere freeze-dried preparation and a preparation method thereof

Abstract

The application provides a levothyroxine sodium sustained-release microsphere freeze-drying preparation for injection, which comprises 10-20 parts by weight of levothyroxine sodium, 30-60 parts by weight of polylactic acid-glycolic acid copolymer, 10-20 parts by weight of polyvinyl alcohol and 2-4 parts by weight of sodium chloride. The application has the beneficial effects that the application provides a levothyroxine sodium sustained-release microsphere freeze-drying preparation for injection and a preparation method, 1. Through the slow release, daily medication can be avoided, and patients can avoid forgetting to take medicine; 2. The preparation has a good appearance and a short reconstitution time.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of freeze-drying of microspheres, and particularly relates to a levothyroxine sodium sustained-release microsphere freeze-dried preparation for injection and a preparation method thereof. BACKGROUND

[0002] Hypothyroidism is a common clinical symptom of thyroid hormone deficiency, and if not treated in time, it can cause serious adverse health effects to multiple organ systems. Clinically, overt primary hypothyroidism is defined as an elevated concentration of thyroid-stimulating hormone (TSH) and a concentration of free thyroxine (fT4) lower than the reference range. Subclinical hypothyroidism is generally considered an early sign of thyroid dysfunction, and is defined as an elevated concentration of TSH but a concentration of fT4 within the reference range. Depending on the pathology of the thyroid, pituitary or hypothalamus or peripheral tissues, hypothyroidism can be primary, central or peripheral.

[0003] Acquired primary hypothyroidism is the most common form and can be caused by severe iodine deficiency, but more often by chronic autoimmune thyroiditis in iodine-sufficient areas. In most cases, the onset of hypothyroidism is insidious, and symptoms can appear later in the disease process. There is great variability in clinical presentation and symptoms of hypothyroidism, especially in pregnancy and children. In the United States, National Health and Nutrition Examination Survey data show a prevalence of hypothyroidism (clinically overt and occult) of 4.6%. A screening study in the United States showed a prevalence of clinically overt hypothyroidism and subclinical hypothyroidism of 0.4% and 9%, respectively, while the latter increased to more than 20% in women 75 years of age or older. In a meta-analysis from Europe, the prevalence of clinically overt hypothyroidism and subclinical hypothyroidism was 0.37% and 3.8%, respectively, including diagnosed and undiagnosed cases, and the incidence of hypothyroidism was estimated at 226 cases per 100,000 people per year.

[0004] In areas with severe iodine deficiency, universal salt iodization has been successful in reducing hypothyroidism. Nevertheless, iodine deficiency remains an important public health problem. Adequate iodine intake is important for everyone, and iodine deficiency can trigger or exacerbate hypothyroidism. The recommended daily dose of iodine is: 90 μg for preschool children, 120 μg for school-age children, 150 μg for adults, and 250 μg for pregnant women.

[0005] At present, the mainstream drug treatment of hypothyroidism is levothyroxine sodium tablets (L-T4), thyroid tablets [containing triiodothyronine (T3), which is not commonly used in clinical]. But the treatment plan of levothyroxine sodium tablets requires patients to take drugs orally every day. For elderly patients, forgetting to take medicine often occurs, which can cause serious consequences and is not conducive to the rehabilitation of patients. SUMMARY

[0006] In order to solve the above technical problems, the drug administration interval is prolonged, the drug frequency is reduced, the drug compliance of the patient is improved, the toxic side effects caused by oral drugs are avoided, and the stimulation of the gastrointestinal tract is reduced. The present application provides a kind of levothyroxine sodium sustained-release microspheres freeze-dried preparation for injection and a preparation method thereof.

[0007] The specific technical scheme of the present application is as follows:

[0008] In one aspect, the present application provides a kind of levothyroxine sodium sustained-release microspheres freeze-dried preparation for injection, comprising: levothyroxine sodium 10-20 parts by weight, poly (lactic-co-glycolic acid) 30-60 parts by weight, polyvinyl alcohol 10-20 parts by weight, and sodium chloride 2-4 parts by weight.

[0009] Further, it also includes 0.1-10 parts by weight of a freeze-drying protective agent, preferably, it includes 5 parts by weight of a freeze-drying protective agent.

[0010] Further, the freeze-drying protective agent includes sodium citrate, trehalose and mannitol, preferably, the freeze-drying protective agent includes 2 parts by weight of sodium citrate, 2 parts by weight of trehalose and 1 part by weight of mannitol.

[0011] Further, the poly (lactic-co-glycolic acid) includes poly (lactic-co-glycolic acid) (75:25) and poly (lactic-co-glycolic acid) (50:50).

[0012] Further, the poly (lactic-co-glycolic acid) includes 15-30 parts by weight of poly (lactic-co-glycolic acid) (75:25) and 15-30 parts by weight of poly (lactic-co-glycolic acid) (50:50), preferably, the poly (lactic-co-glycolic acid) includes 25 parts by weight of poly (lactic-co-glycolic acid) (75:25) and 25 parts by weight of poly (lactic-co-glycolic acid) (50:50).

[0013] In another aspect, the present application also provides a preparation method of levothyroxine sodium sustained-release microspheres freeze-dried preparation for injection, comprising:

[0014] S1: dissolving levothyroxine sodium, poly (lactic-co-glycolic acid), polyvinyl alcohol and sodium chloride in water to obtain a solution;

[0015] S2: centrifuging the solution obtained in step S1 at 900 rpm for 2 min, discarding the supernatant, washing with water for several times, and obtaining the microspheres by centrifugation;

[0016] S3: adding a certain amount of the microspheres and a lyoprotectant in proportion, stirring, and obtaining a suspension;

[0017] S4: lyophilizing the suspension to obtain a levothyroxine sodium sustained-release microsphere lyophilized preparation.

[0018] Further, the lyophilization process in the S4 step specifically comprises:

[0019] S4-1: pre-freezing the suspension;

[0020] S4-2: sublimating the product obtained in S4-1 once;

[0021] S4-3: resolving and drying the product obtained in S4-2 to obtain a levothyroxine sodium sustained-release microsphere lyophilized preparation for injection.

[0022] Further, in particular, the pre-freezing comprises a first pre-freezing process, a second pre-freezing process, and a third pre-freezing process;

[0023] The parameters of the first pre-freezing process are a temperature of -40°C, a temperature change time of 60 min, and a holding time of 30 min;

[0024] The parameters of the second pre-freezing process are a temperature of -70°C, a temperature change time of 30 min, and a holding time of 60 min;

[0025] The parameters of the third pre-freezing process are a temperature of -50°C, a temperature change time of 30 min, and a holding time of 180 min.

[0026] Further, the sublimation once comprises a first sublimation once process and a second sublimation once process;

[0027] The parameters of the first sublimation once process are a temperature of -15°C, a temperature change time of 120 min, a holding time of 1200 min, and a vacuum degree of 0.01 mbar;

[0028] The parameters of the second sublimation once process are a temperature of 0°C, a temperature change time of 60 min, a holding time of 360 min, and a vacuum degree of 0.01 mbar.

[0029] Further, the resolving and drying comprises a first resolving and drying process and a second resolving and drying process;

[0030] The parameters of the first resolving and drying process are a temperature of 20°C, a temperature change time of 240 min, a holding time of 120 min, and a vacuum degree of 0.01 mbar.

[0031] The parameters of the second analysis drying process are: temperature is 30 DEG C, temperature changing time is 60 min, holding time is 240 min, vacuum degree is 0.01 mbar.

[0032] The present application has the advantages that the present application provides a levothyroxine sodium for injection sustained-release microspheres lyophilized preparation and preparation method, 1. Through the slow release, the daily medication can be avoided, the patient can be avoided to forget to take the medicine, and the continuous action time reaches 2-4 weeks; 2. The appearance of the preparation is good, and the reconstitution time is short. DETAILED DESCRIPTION

[0033] The following examples further illustrate the present application, but should not be construed as limiting the present application. Modifications or replacements of the method, steps or conditions of the present application without departing from the spirit and essence of the present application shall fall within the scope of the present application.

[0034] The raw and auxiliary materials used in the present application are commercially available, and the specific sources are as follows:

[0035] Levothyroxine sodium: Shenzhen Zhonglian Pharmaceutical Co., Ltd.

[0036] Polylactic acid-glycolic acid copolymer: Evonik

[0037] Polyvinyl alcohol: Evonik

[0038] Sodium chloride: Hunan Erkang Pharmaceutical Co., Ltd.

[0039] Sodium citrate: Hunan Erkang Pharmaceutical Co., Ltd.

[0040] Trehalose: Hunan Erkang Pharmaceutical Co., Ltd.

[0041] Mannitol: Hunan Erkang Pharmaceutical Co., Ltd.

[0042] Example 1: The prescription of the levothyroxine sodium for injection sustained-release microspheres lyophilized preparation is shown in Table 1:

[0043] Table 1. The prescription of the levothyroxine sodium for injection sustained-release microspheres lyophilized preparation

[0044]

[0045] The preparation process is:

[0046] S1: Levothyroxine sodium, polylactic acid-glycolic acid copolymer, polyvinyl alcohol and sodium chloride are dissolved in water to obtain a solution;

[0047] S2: The solution obtained in step S1 is centrifuged at 900 rpm for 2 min, the supernatant is discarded, and the microspheres are obtained after washing with water twice and centrifugation;

[0048] S3: Take half of the microspheres, add the freeze-drying protectant according to the prescription ratio, stir to obtain a suspension;

[0049] S4: The suspension is filled into 7ml borosilicate vials, with a filling amount of 2ml, and freeze-dried to obtain a levothyroxine sodium sustained-release microsphere freeze-dried preparation;

[0050] The freeze-drying process is as follows:

[0051] S4-1: The suspension is pre-frozen;

[0052] S4-2: The product obtained in S4-1 is subjected to primary sublimation;

[0053] S4-3: The product obtained in S4-2 is subjected to analytical drying to obtain a levothyroxine sodium sustained-release microsphere freeze-dried preparation for injection.

[0054] The pre-freezing includes a first pre-freezing process, a second pre-freezing process and a third pre-freezing process;

[0055] The parameters of the first pre-freezing process are a temperature of -40℃, a temperature change time of 60min, and a holding time of 30min;

[0056] The parameters of the second pre-freezing process are a temperature of -70℃, a temperature change time of 30min, and a holding time of 60min;

[0057] The parameters of the third pre-freezing process are a temperature of -50℃, a temperature change time of 30min, and a holding time of 180min.

[0058] The primary sublimation includes a first primary sublimation process and a second primary sublimation process;

[0059] The parameters of the first primary sublimation process are a temperature of -15℃, a temperature change time of 120min, a holding time of 1200min, and a vacuum degree of 0.01mbar;

[0060] The parameters of the second primary sublimation process are a temperature of 0℃, a temperature change time of 60min, a holding time of 360min, and a vacuum degree of 0.01mbar.

[0061] The analytical drying includes a first analytical drying process and a second analytical drying process;

[0062] The parameters of the first analytical drying process are a temperature of 20℃, a temperature change time of 240min, a holding time of 120min, and a vacuum degree of 0.01mbar;

[0063] The parameters of the second analytical drying process are a temperature of 30℃, a temperature change time of 60min, a holding time of 240min, and a vacuum degree of 0.01mbar.

[0064] Example 2, except that levothyroxine sodium 15 parts by weight, polylactic acid- glycolic acid copolymer (75:25) 25 parts by weight, polylactic acid-glycolic acid copolymer (50:50) 25 parts by weight, polyvinyl alcohol 15 parts by weight, sodium chloride 3 parts by weight.

[0065] Example 3, except that levothyroxine sodium 20 parts by weight, polylactic acid- glycolic acid copolymer (75:25) 30 parts by weight, polylactic acid-glycolic acid copolymer (50:50) 30 parts by weight, polyvinyl alcohol 20 parts by weight, sodium chloride 4 parts by weight.

[0066] Comparative Example 1, except that polylactic acid-glycolic acid copolymer (75:25) 30 parts by weight, polylactic acid-glycolic acid copolymer (50:50) is not used.

[0067] Comparative Example 2, except that polylactic acid-glycolic acid copolymer (50:50) 30 parts by weight, polylactic acid-glycolic acid copolymer (75:25) is not used.

[0068] Comparative Example 3, except that polylactic acid-glycolic acid copolymer (75:25) 5 parts by weight, polylactic acid-glycolic acid copolymer (50:50) 5 parts by weight.

[0069] Comparative Example 4, except that polylactic acid-glycolic acid copolymer (75:25) 40 parts by weight, polylactic acid-glycolic acid copolymer (50:50) 40 parts by weight.

[0070] Example 4, except that the formulation is as shown in Table 1:

[0071] Table 2. Formulation of levothyroxine sodium sustained-release microspheres for injection

[0072]

[0073] Example 5: except that sodium citrate 0.04 parts by weight, trehalose 0.04 parts by weight, mannitol 0.02 parts by weight.

[0074] Example 6: except that sodium citrate 4 parts by weight, trehalose 4 parts by weight, mannitol 2 parts by weight.

[0075] Comparative Example 5: except that sodium citrate 0.02 parts by weight, trehalose 0.03 parts by weight, mannitol 0.02 parts by weight.

[0076] Comparative Example 6: except that sodium citrate 10 parts by weight, trehalose 5 parts by weight, mannitol 5 parts by weight.

[0077] Comparative Example 7: Different from Example 4, trehalose and mannitol were not used.

[0078] Comparative Example 8: Different from Example 4, sodium citrate, trehalose and mannitol were replaced by glycine 5 parts by weight.

[0079] Comparative Example 9: Different from Example 4, sodium citrate, trehalose and mannitol were replaced by gelatin 5 parts by weight.

[0080] Test Example 1: Drug release rate test: Take Example 1, Example 2,

[0081] Experimental reagent: PBS solution

[0082] Experimental instrument: High performance liquid chromatograph, vortex, centrifuge, water bath shaker

[0083] Experimental method: Precisely take 10 mg of drug-containing microspheres into a 10 mL centrifuge tube, add 10 mL of buffer solution, and place it in a constant temperature water bath shaker. The in vitro release rate of the microspheres was determined at a shaking speed of 100 rpm and a temperature of 37°C ± 0.5°C.

[0084] Sampling method: Take out 5 mL of solution at 0.5, 1, 1.5, 2, 4, 6, 8, 12, 24, 36, 48, 60, 72, 84, 96, 120 h, and supplement with the same volume of PBS solution. The results are shown in Table 3.

[0085] Table 3. Drug release rate

[0086]

[0087]

[0088] As can be seen from Table 3, the release rate of the products of Examples 1-3 is significantly better than that of Comparative Examples 1-2. Comparative Example 1 cannot be completely released within 1 month, and Comparative Example 2 is released too quickly and is completely released in 2 weeks. This shows that the ratio of polylactic acid to hydroxyacetic acid in polylactic acid-hydroxyacetic acid copolymer will affect the release of the product. The use of polylactic acid-hydroxyacetic acid copolymer (75:25) and polylactic acid-hydroxyacetic acid copolymer (50:50) will have a better release rate.

[0089] The release rate of the products of Examples 1-3 was significantly better than that of Comparative Examples 3-4. Comparative Example 3 released too quickly, and Comparative Example 4 released too slowly to be completely released in one month. The weight ratio of polylactic acid-glycolic acid copolymer (75:25) and polylactic acid-glycolic acid copolymer (50:50) affected the release rate of the product, and the release rate was better when the polylactic acid-glycolic acid copolymer was 30-60 parts by weight.

[0090] Test Example 2: Appearance Evaluation

[0091] The appearance of the products of Examples 1, 4-6 and Comparative Examples 5-9 was evaluated, and the results are shown in Table 3.

[0092] Table 4. Appearance Evaluation Table and Reconstitution Time

[0093]

[0094] As shown in Table 4, the product of Example 1 had good appearance and reconstitution time of less than 1 minute, but was severely damaged under a microscope. Examples 4-6 were better than Example 1, indicating that the freeze-drying protective agent affected the appearance under a microscope after reconstitution, and the appearance under a microscope after reconstitution was better when a freeze-drying protective agent was used.

[0095] The appearance and reconstitution time of less than 1 minute of Examples 4-6 were better than those of Comparative Examples 7-9, which had damaged appearance and long reconstitution time, indicating that the selection of the freeze-drying protective agent affected the appearance and reconstitution time of the product. When sodium citrate, trehalose and mannitol were used as the freeze-drying protective agent, the appearance of the product was better and the reconstitution time was shorter.

[0096] The appearance and reconstitution time of less than 1 minute of Examples 4-6 were better than those of Comparative Examples 5, 6-7. Comparative Example 5 had damaged appearance although the reconstitution time was less than 1 minute. Comparative Example 6 had good appearance and reconstitution time of less than 1 minute, but was damaged under a microscope after reconstitution. Comparative Example 7 had damaged appearance and reconstitution time of more than 1 minute, indicating that the weight ratio of the freeze-drying protective agent affected the appearance and reconstitution time of the product. When sodium citrate, trehalose and mannitol were used as the freeze-drying protective agent and the weight ratio was 0.1-10, the appearance of the product was better and the reconstitution time was shorter.

Claims

1. A lyophilized formulation of levothyroxine sodium sustained-release microspheres for injection, characterized in that, Comprise: Levothyroxine sodium 10 parts by weight, polylactic acid-glycolic acid copolymer 75:25 15 parts by weight, polylactic acid-glycolic acid copolymer 50:50 15 parts by weight, polyvinyl alcohol 10 parts by weight and sodium chloride 2 parts by weight; also comprising: sodium citrate 2 parts by weight, trehalose 2 parts by weight, mannitol 1 part by weight, or sodium citrate 0.04 parts by weight, trehalose 0.04 parts by weight, mannitol 0.02 parts by weight; or sodium citrate 4 parts by weight, trehalose 4 parts by weight, mannitol 2 parts by weight.

2. A process for the preparation of a lyophilized formulation of levothyroxine sodium sustained release microspheres for injection as claimed in claim 1, characterized in that, Comprise: S1: levothyroxine sodium, polylactic acid-glycolic acid copolymer 75:25, polylactic acid-glycolic acid copolymer 50:50, polyvinyl alcohol and sodium chloride are dissolved in water to obtain a solution; S2: the solution obtained in step S1 is centrifuged at 900 rpm for 2 min, the supernatant is discarded, and the microspheres are obtained after washing with water several times and centrifugation; S3: a certain amount of microspheres is added with sodium citrate, trehalose and mannitol in proportion, stirred to obtain a suspension; S4: the suspension is freeze-dried to obtain levothyroxine sodium sustained-release microspheres freeze-dried preparation.

3. The method of claim 2, wherein the method is characterized by, The freeze-drying process in the S4 step specifically comprises: S4-1: pre-freezing the suspension; S4-2: sublimating the product obtained in S4-1 once; S4-3: resolving and drying the product obtained in S4-2 to obtain levothyroxine sodium sustained-release microspheres freeze-dried preparation for injection.

4. The method of claim 3, wherein the method is characterized by, The pre-freezing includes a first pre-freezing process, a second pre-freezing process and a third pre-freezing process; The parameters of the first pre-freezing process are temperature -40℃, temperature changing time 60min, and holding time 30min; The parameters of the second pre-freezing process are temperature -70℃, temperature changing time 30min, and holding time 60min; The parameters of the third pre-freezing process are temperature -50℃, temperature changing time 30min, and holding time 180min.

5. The method of claim 3, wherein the method is characterized by the steps of: The sublimation once includes a first sublimation once process and a second sublimation once process; The parameters of the first sublimation once process are temperature -15℃, temperature changing time 120min, holding time 1200min, and vacuum degree 0.01mbar; The parameters of the second sublimation once process are temperature 0℃, temperature changing time 60min, holding time 360min, and vacuum degree 0.01mbar.

6. The method of claim 3, wherein the method is characterized by the steps of: The resolving and drying includes a first resolving and drying process and a second resolving and drying process; The parameters of the first resolving and drying process are temperature 20℃, temperature changing time 240min, holding time 120min, and vacuum degree 0.01mbar; The parameters of the second resolving and drying process are temperature 30℃, temperature changing time 60min, holding time 240min, and vacuum degree 0.01mbar.

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