Vitamin D2 injection composition with improved formula and preparation method thereof

Abstract

The invention provides a vitamin D2 injection composition with an improved formula. The composition comprises 22.5-15% w / v of vitamin D2, 6-12% w / v of a cosolvent, 3-5% w / v of an isotonic agent, 0-2% w / v of a pH regulator and the balance of water for injection, wherein the cosolvent is at least one of medium chain triglyceride, Tween 20, Tween 40 and Tween 80. The composition is reasonable in formula, simple in process and good in stability, can be used for preparing a vitamin D2 water-soluble injection, and has remarkable advantages compared with an oily vitamin D2 preparation which is high in irritation and prone to allergy.

Description

Technical Field

[0001] The present application belongs to the field of pharmaceutical technology. Specifically, the present application provides a vitamin D2 injection composition with an improved formula and a preparation method. Background Art

[0002] Vitamin D2, also known as ergocalciferol, is a key member of the vitamin D family. It promotes calcium and phosphorus absorption by intestinal mucosal cells and is an important treatment for rickets in children and osteoporosis in the elderly. Vitamin D is unstable and sensitive to light, heat, and oxidation. It can transform into many chemically similar isomers. However, with the exception of provitamin D, most other isomers and impurities are inactive. Vitamin D2 is produced from ergosterol found in plants or yeast through ultraviolet irradiation. Industrial production in my country began in the 1980s.

[0003] Vitamin D2 is extremely unstable when exposed to light, heat, and air, and its water insolubility limits its widespread use in pharmaceuticals, food, and feed. Precisely because of its unstable chemical properties, vitamin D2 cannot be used clinically without the establishment of a stable formulation production system. Numerous methods have been proposed. JP57-45415, JP61-41351, JP62-51948, JP58-206533, and US Pat. No. 4,729,895 disclose methods for stabilizing oral formulations. Soft capsules and tablets prepared using these techniques, marketed under the trade names DRISDOL and VITAMIN D, have found practical clinical application. However, these techniques only address the stabilization of non-aqueous liquid or solid formulations. Furthermore, there is a strong clinical need for aqueous formulations, particularly stable aqueous solutions of vitamin D2. Currently, no research has reported on the administration of stable aqueous solutions as injectable formulations, thus creating an unmet need. Ethyl oleate-containing injectable oil solutions are marketed clinically under the trade names CALCIFEROL BBP and STEROGYL 15H, respectively.

[0004] US20140187520, a stable injectable pharmaceutical composition of a vitamin D receptor agonist and its preparation method; US20100075933, an injectable composition of a vitamin D compound.

[0005] There appears to be a delicate balance between pH, solubility, and stability of vitamin D2 in an aqueous environment, where achieving this balance and developing shelf-stable aqueous formulations is a formidable challenge. Therefore, a shelf-stable and effective solution formulation is highly desirable to prevent degradation of the therapeutically active entity and maintain the drug in solution at the desired pH, and considerable effort has been devoted to achieving this goal. Summary of the Invention

[0006] In response to the above situation, on the one hand, the present application provides a vitamin D2 injection composition with an improved formula, which comprises 2.5-15% w / v of vitamin D2, 6-12% w / v of a cosolvent, 3-5% w / v of an isotonic agent, 0-2% w / v of a pH regulator, and the balance is supplemented by water for injection; wherein the cosolvent is at least one of medium-chain triglycerides, Tween 20, Tween 40, and Tween 80.

[0007] Furthermore, the cosolvent is medium chain triglyceride and Tween 80.

[0008] Furthermore, the composition comprises 5-10% w / v of Tween 80; and the mass ratio of Tween 80 to medium-chain triglycerides in the composition is 5-12:1.

[0009] Furthermore, the mass ratio of Tween 80 to medium chain triglycerides in the composition is 5:1.

[0010] Furthermore, the isotonic agent is at least one of 70% crystalline sorbitol solution, xylitol, lactose, galactose, and glucose.

[0011] Furthermore, the isotonic agent is a 70% crystalline sorbitol solution.

[0012] Furthermore, the pH adjuster is at least one of sodium hydroxide, sodium dihydrogen phosphate dihydrate, disodium hydrogen phosphate, hydrochloric acid, and sodium citrate.

[0013] Furthermore, the pH regulator is sodium hydroxide and sodium dihydrogen phosphate dihydrate.

[0014] Furthermore, the pH of the composition is 3.5 to 6.5.

[0015] The composition provided by the present invention is stored at 40° C. and 75% relative humidity for 6 months. The vitamin D2 impurity D in the injection is not detected, and no new unknown impurities are generated that affect the quality of the raw material drug. The chemical formula of the vitamin D2 impurity D is:

[0016]

[0017] On the other hand, the present application provides the use of the above-mentioned composition in the preparation of vitamin D2 injection.

[0018] Furthermore, the injection is a subcutaneous, intravenous or intramuscular injection, preferably a subcutaneous injection.

[0019] Furthermore, the injection is placed in a sealed container, and the sealed container is selected from the group consisting of an ampoule, a vial and a pre-filled syringe.

[0020] On the other hand, the present application provides a method for preparing the above composition, the method comprising:

[0021] 1) Add 80% water for injection into the liquid preparation tank, adjust the water temperature to 40-50°C, and fill with nitrogen for 30 minutes;

[0022] 2) Protect from light, add the prescribed amount of excipients and medium-chain triglycerides to the liquid preparation tank, stir evenly, add the prescribed amount of vitamin D2 and stir until dissolved;

[0023] 3) Add 1) to the solution in 2) while stirring. After the solution changes from a gel-like state to a clear solution, add 70% crystalline sorbitol solution and stir to mix. Adjust the pH to 3.5-6.5 and add water for injection to the full volume.

[0024] Furthermore, the method further comprises:

[0025] 4) Detect and determine the content and pH value of the intermediate. If qualified, filter with a microporous membrane;

[0026] 5) Canning and sealing.

[0027] Furthermore, in step 4), the intermediate content and pH value are detected and determined. After passing the test, the product is filtered through a 0.22 μm microporous membrane under a nitrogen atmosphere.

[0028] Furthermore, in step 5), the mixture is filled into 1 ml brown ampules, with each ampoule filled with 1 ml. Before sealing, filtered nitrogen is introduced into the head space of each bottle; the mixture is labeled, packaged, and inspected to obtain the finished product.

[0029] On the other hand, the present application also provides the use of the above-mentioned composition in the preparation of drugs or health products for treating vitamin D deficiency or calcium supplementation.

[0030] On the other hand, the present application also provides the use of the above-mentioned composition in the preparation of drugs for treating vitamin D-dependent rickets, hypophosphatemia, hypoparathyroidism, renal insufficiency, and renal bone atrophy.

[0031] The vitamin D2 injection of the present invention can be administered by self-subcutaneous injection at home after receiving guidance; water-soluble preparations reduce viscosity and absorb more completely, reduce pain, the risk of nodules, and have advantages in convenience and comfort; there is obvious clinical advantage compared to the vitamin D2 injection (oil solution) currently on the market. The pain reaction is large, irritating, allergic and easily causes drug accumulation in local muscle tissue to cause lump pain when injected on the market, while the vitamin D2 injection (aqueous solution) prescription does not have solvents and preservatives (the oil solution solvent is sesame oil, containing the preservative benzyl alcohol) that may cause severe reactions or abscesses. In addition, the subcutaneous injection mode of administration can reduce the occurrence of multiple adverse reactions, and the patient can inject it by himself at the same time, while greatly improving convenience, reducing the additional cost of treatment, and effectively improving the acceptance and compliance of the patient.

[0032] The aqueous vitamin D2 injection of the present invention has a reasonable formulation, simple process, and good stability. After a 6-month accelerated test at 40°C, no significant changes in the relevant substances were observed. Further features and advantages of the present invention will become apparent from the detailed description below. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 These are chromatograms of multiple formulas, from bottom to top, they are formula 20, formula 21, formula 22, formula 23, vitamin D2 raw material, and formula 1. DETAILED DESCRIPTION

[0034] The following includes the implementation of the preferred embodiment to illustrate the practice of the present invention. It is understood that the details shown are for illustrative discussion of the preferred embodiments of the present invention by way of example and purpose, and are intended to demonstrate what is considered to be the most useful and easy-to-understand description of the procedures for making the invention and the principles and concepts. It is obvious to those skilled in the art that the present invention is not limited to the details of the following illustrative examples, and that the invention can be embodied in other specific forms without departing from its essential attributes. Therefore, it is intended that the present embodiments and examples be considered in all aspects as illustrative and not restrictive, and that all changes that fall within the equivalent meaning and scope of the claims should be included therein.

[0035] Example 1 Detection method:

[0036] The detection conditions and impurity types of the HPLC method for effectively detecting impurities in vitamin D2 injection are shown in Tables 1 and 2:

[0037] Table 1 HPLC test conditions for detection and quantification of impurities in vitamin D2 injection

[0038]

[0039] Table 2 Relative retention time (RRT) of each impurity phase of vitamin D2

[0040]

[0041]

[0042] Example 2 Establishment of an effective solubilizing agent dissolution system:

[0043] Because the API has poor solubility in water, in order to ensure the development of subsequent formulation process parameters, we screened the excipients in the prescription, with the aim of screening out suitable solvents / cosolvents to increase the solubility of the API and enable the API to be completely dissolved in the prescription.

[0044] Commonly used enhancers / solvents in injections were selected, such as polyethylene glycol-15-hydroxystearate, Tween 80, lecithin, poloxamer 188, ethanol, propylene glycol, polyethylene glycol 400, and hydroxypropyl beta-cyclodextrin. Each excipient was added in the usual amount. According to the proposed prescription composition, each excipient was weighed into a suitable container, water was added to the full amount of the prescription, and the mixture was stirred to dissolve. Then, the prescribed amount of vitamin D2 was added and stirred to dissolve. The dissolution phenomenon of vitamin D2 and the properties of the drug solution were observed during the test. The prescription composition is shown in Table 3, and the test results are shown in Table 4.

[0045] Table 3 Prescription composition

[0046]

[0047] Table 4 Screening results of the dosage of each excipient

[0048]

[0049] Conclusion: The above experimental phenomena and results indicate that, among the excipients mentioned above, Tween 80 has a significant solubility-enhancing effect. Lecithin also has a relatively significant solubility-enhancing effect, but bubbles are present. Formulation 5 dissolves very quickly. Based on the experimental results, Tween 80 and medium-chain triglycerides are preferred as solubilizers and cosolvents in this product formulation, and further investigation is needed to determine the optimal dosage.

[0050] Example 3 Screening of antioxidants:

[0051] Since vitamin D2 is extremely unstable to oxygen, it is generally understood that aqueous solutions, especially injection solutions, must be supplemented with appropriate antioxidants. Representative antioxidants available for injection in clinical injection solutions include sodium metabisulfite, sodium sulfite, sodium bisulfite, disodium edetate dihydrate, and vitamin C. Our detailed investigation results are shown in Table 5:

[0052] Table 5 Antioxidant investigation results

[0053]

[0054]

[0055] Conclusion: It can be seen that the addition of antioxidants cannot bring stability to this product, which is an unexpected result.

[0056] Example 4 Investigation and Optimization of the Dosage of Tween 80 / Medium Chain Triglycerides:

[0057] According to the results of the excipient type screening, different dosages of Tween 80 / medium-chain triglycerides were designed for sample preparation based on prescriptions 2 and 5, and the dosage of Tween 80 was optimized. During the test, the dissolution time, dissolution phenomenon, properties of the drug solution, pH value and content of each prescription were focused on. The prescription composition and test results are shown in Table 6.

[0058] Table 6 Results of the investigation on the dosage of Tween 80 / medium chain triglycerides

[0059]

[0060] The dosage of Tween 80 in the above prescription ranges from 5% to 10% (w / v). As the dosage of Tween 80 increases, the dissolution rate becomes faster. In addition, medium-chain triglycerides have extremely high solubility for vitamin D2, but medium-chain triglycerides are insoluble in water. The added amount has extremely high requirements on the ratio of the clarified injection. If more than a little is added, the injection will stratify or emulsify. After a large number of proportion studies, the dosage of polysorbate 80 (Tween 80) is 5% to 10% (w / v) which is the most reasonable. A large amount will increase the toxicity, and a small amount cannot be made into an injection. The weight ratio of medium-chain triglycerides to polysorbate 80 (Tween 80) is at least 5:1. The optimal ratio varies according to the dosage of vitamin D2, with a maximum of 12:1.

[0061] Example 5 Selection of isotonicity regulator:

[0062] Commonly used injection isotonicity regulators include crystalline sorbitol solution (70%), xylitol, lactose, glucose, and sodium chloride. The screening results are shown in Table 7:

[0063] Table 7 Isotonicity Regulator Investigation Results

[0064]

[0065]

[0066] The results showed that crystalline sorbitol solution (70%) was very stable as an isotonicity regulator.

[0067] Formulations 1-4: Preparation of Vitamin D2 Injection (Percentages are w / v, weight / volume, the same below)

[0068] prescription:

[0069]

[0070] Preparation process:

[0071] 1) Add 80% water for injection into the liquid preparation tank, adjust the water temperature to 40-50°C, and fill with nitrogen for 30 minutes;

[0072] 2) Protect from light, add the prescribed amount of excipients and medium-chain triglycerides to the liquid preparation tank, stir evenly, add the prescribed amount of vitamin D2 and stir until dissolved;

[0073] 3) Add 1) to the solution in 2) while stirring. After the solution changes from a gel to a clear solution, add crystalline sorbitol solution (70%) and stir to mix. Adjust the pH to 3.5-6.5 and add water for injection to the full volume. Determine the content of the intermediate and the pH value. If qualified, filter through a 0.22 μm microporous membrane under a nitrogen atmosphere.

[0074] 4) Fill the product into 1 ml brown ampoules, filling each bottle with 1 ml. Before sealing, introduce filtered nitrogen into the head space of each bottle; label and package, and obtain the finished product after passing the inspection.

[0075] Recipes 5-8:

[0076] prescription:

[0077]

[0078] Preparation process:

[0079] 1) Add 80% water for injection into the liquid preparation tank, adjust the water temperature to 40-50°C, and fill with nitrogen for 30 minutes;

[0080] 2) Protect from light, add the prescribed amount of excipients and medium-chain triglycerides to the liquid preparation tank, stir evenly, add the prescribed amount of vitamin D2 and stir until dissolved;

[0081] 3) Add 1) to the solution in 2) while stirring. After the solution changes from a gel to a clear solution, add crystalline sorbitol solution (70%) and stir to mix. Adjust the pH to 3.5-6.5 and add water for injection to the full volume. Determine the content of the intermediate and the pH value. If qualified, filter through a 0.22 μm microporous membrane under a nitrogen atmosphere.

[0082] 4) Fill the product into 1 ml brown ampoules, filling each bottle with 1 ml. Before sealing, introduce filtered nitrogen into the head space of each bottle; label and package, and obtain the finished product after passing the inspection.

[0083] Recipes 9-12:

[0084] prescription:

[0085]

[0086] Preparation process:

[0087] 1) Add 80% water for injection into the liquid preparation tank, adjust the water temperature to 40-50°C, and fill with nitrogen for 30 minutes;

[0088] 2) Protect from light, add the prescribed amount of excipients and medium-chain triglycerides to the liquid preparation tank, stir evenly, add the prescribed amount of vitamin D2 and stir until dissolved;

[0089] 3) Add 1) to the solution in 2) while stirring. After the solution changes from a gel to a clear solution, add crystalline sorbitol solution (70%) and stir to mix. Adjust the pH to 3.5-6.5 and add water for injection to the full volume. Determine the content of the intermediate and the pH value. If qualified, filter through a 0.22 μm microporous membrane under a nitrogen atmosphere.

[0090] 4) Fill the product into 1 ml brown ampoules, filling each bottle with 1 ml. Before sealing, introduce filtered nitrogen into the head space of each bottle; label and package, and obtain the finished product after passing the inspection.

[0091] Recipes 13-16:

[0092] prescription:

[0093]

[0094] Preparation process

[0095] 1) Add 80% water for injection into the liquid preparation tank, adjust the water temperature to 40-50°C, and fill with nitrogen for 30 minutes;

[0096] 2) Protect from light, add the prescribed amount of excipients and medium-chain triglycerides to the liquid preparation tank, stir evenly, add the prescribed amount of vitamin D2 and stir until dissolved;

[0097] 3) Add 1) to the solution in 2) while stirring. After the solution changes from a gel to a clear solution, add crystalline sorbitol solution (70%) and stir to mix. Adjust the pH to 3.5-6.5 and add water for injection to the full volume. Determine the content of the intermediate and the pH value. If qualified, filter through a 0.22 μm microporous membrane under a nitrogen atmosphere.

[0098] 4) Fill the product into 1 ml brown ampoules, filling each bottle with 1 ml. Before sealing, introduce filtered nitrogen into the head space of each bottle; label and package, and obtain the finished product after passing the inspection.

[0099] Comparative Formulas 17-20: Combination Effects of Other Vitamin D Compounds and Tween 80 / Medium Chain Triglycerides:

[0100] To expand the API of the above preferred formulation, the applicant tried a variety of vitamin D substances used clinically for injection. According to the present invention, the optimal ratio of Tween 80 / medium chain triglycerides is 5:1, and injections with a specification of 1 ml:5 mg were prepared. The preparation method is the same as above. The results are shown in Table 8:

[0101] Table 8 Effects of other vitamin D compound injection combinations

[0102]

[0103] The prescription of Doxil Injection in the United States is shown in Table 9:

[0104] Table 9 Prescription of HECTOROL Injection

[0105]

[0106] 2 μg of API of doxorcalciferol requires Tween 20 to be 10 mg, and 0.05 ml of toxic and volatile ethanol is also used for solubilization. This shows that other vitamin D compounds cannot achieve the effects of the present invention.

[0107] Comparative formula 20-23: Effects of other organic solvent-containing co-solvent combinations:

[0108] 1. Prescription (to dissolve organic cosolvents in excess of the conventional dosage for injection)

[0109]

[0110]

[0111] Preparation method: similar to Example 1.

[0112] Example 6 Stability Study:

[0113] Typical injection solutions prepared from the formula samples and the comparative formula samples were placed in a constant temperature and humidity incubator, set at 75% ± 5% humidity and 40°C ± 2°C, for a 6-month accelerated test. Sampling was performed at the end of each month. The results of the relevant substances are shown in Table 10:

[0114]

[0115]

[0116] Conclusion: The composition of the present invention is extremely stable under accelerated test.

Claims

1. A vitamin D2 injection composition with an improved formula, characterized in that: The composition comprises 2.5-15% w / v of vitamin D2, 6-12% w / v of a cosolvent, 3-5% w / v of an isotonic agent, 0-2% w / v of a pH regulator, and the remainder is supplemented by water for injection; wherein the cosolvent is at least one of medium-chain triglycerides, Tween 20, Tween 40, and Tween 80.

2. The composition according to claim 1, wherein the cosolvent is medium chain triglyceride and Tween 80. 3 . The composition according to claim 2 , comprising 5-10% w / v Tween 80; the mass ratio of Tween 80 to medium-chain triglycerides in the composition is 5-12:

1. 4 . The composition according to claim 3 , wherein the mass ratio of Tween 80 to medium chain triglycerides in the composition is 5:

1.

5. The composition according to claim 1, wherein the isotonic agent is at least one of 70% crystalline sorbitol solution, xylitol, lactose, galactose, and glucose. The composition according to claim 5 , wherein the isotonic agent is a 70% crystalline sorbitol solution.

7. The composition according to claim 1, wherein the pH regulator is at least one of sodium hydroxide, sodium dihydrogen phosphate dihydrate, disodium hydrogen phosphate, hydrochloric acid, and sodium citrate.

8. The composition according to claim 7, wherein the pH adjusting agent is sodium hydroxide and sodium dihydrogen phosphate dihydrate.

9. The composition according to claim 1, wherein the pH of the composition is 3.5 to 6.

5.

10. The composition according to claim 1, wherein the vitamin D2 impurity D is not detected when the composition is stored at 40° C. and 75% relative humidity for 6 months, and the vitamin D2 impurity D has the chemical formula:

11. Use of the composition according to any one of claims 1 to 10 in the preparation of vitamin D2 injection. The use according to claim 11 , wherein the injection is a subcutaneous, intravenous or intramuscular injection.

13. The use according to claim 12, wherein the injection is a subcutaneous injection.

14. The use according to any one of claims 11 to 13, wherein the injection is placed in a sealed container, and the sealed container is selected from the group consisting of an ampoule, a vial and a prefilled syringe.

15. A method for preparing a composition according to any one of claims 1 to 10, comprising: 1) Add water for injection into the liquid preparation tank, adjust the water temperature to 40-50°C, and fill with nitrogen for standby use; 2) Protect from light, add the prescribed amount of excipients and medium-chain triglycerides to the liquid preparation tank, stir evenly, add the prescribed amount of vitamin D2 and stir until dissolved; 3) Add 1) to the solution in 2) while stirring. After the solution changes from a gel-like state to a clear solution, add 70% crystalline sorbitol solution and stir to mix. Adjust the pH to 3.5-6.5 and add water for injection to the full volume.

16. The method according to claim 15, further comprising: 4) Detect and determine the content and pH value of the intermediate. If qualified, filter with a microporous membrane; 5) Canning and sealing.

17. The method according to claim 16, wherein in step 4), the intermediate content and pH value are detected and determined, and after passing the test, the product is filtered through a microporous membrane under a nitrogen atmosphere.

18. The method according to claim 17, wherein in step 5), the solution is filled into brown ampoules at a filling volume of 1 ml per bottle, and filtered nitrogen is introduced into the headspace of each bottle before sealing; the solution is labeled, packaged, and inspected to obtain the finished product.

19. Use of the composition according to any one of claims 1 to 10 in the preparation of a medicine or health product for treating vitamin D deficiency or supplementing calcium.

20. Use of the composition according to any one of claims 1 to 10 in the preparation of a medicament for treating vitamin D-dependent rickets, hypophosphatemia, hypoparathyroidism, renal insufficiency, and renal bone atrophy.

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