Vitamin d nanomicellar oral solution and methods of making the same

Abstract

The application relates to the technical field of pharmaceutical preparations, in particular to a vitamin D nanomicelle preparation and a preparation method thereof. The vitamin D nanomicelle preparation comprises main material vitamin D and auxiliary materials, the auxiliary materials comprise 15-hydroxyl stearyl acid polyethylene glycol ester and glycocholic acid, the weight ratio of the vitamin D, the 15-hydroxyl stearyl acid polyethylene glycol ester and the glycocholic acid is 1:(5-20):(2-10). In the solution of the vitamin D nanomicelle preparation, 1 mL of the solution comprises 800 IU of vitamin D, 100 mcg-400 mcg of 15-hydroxyl stearyl acid polyethylene glycol ester, 40 mcg-200 mcg of glycocholic acid and a solvent. The vitamin D nanomicelle preparation has good stability, is simple to prepare, has a wide application range and has industrial production value.

Description

Technical Field

[0001] This application relates to the field of pharmaceutical preparation technology, specifically to a vitamin D nanomicelle oral solution and its preparation method. Background Technology

[0002] The main functions of vitamin D include promoting the absorption of calcium and phosphorus in the intestine, maintaining serum calcium and phosphorus stability by inhibiting the release of parathyroid hormone (PTH), ensuring proper bone mineralization, and reducing calcium excretion from the kidneys. Vitamin D acts on the intestines, kidneys, and bones to regulate calcium and phosphorus metabolism. In the small intestine, vitamin D promotes the absorption of calcium and phosphorus. In the kidneys, vitamin D increases the reabsorption of calcium and phosphorus by the renal tubules to maintain normal blood calcium and phosphorus levels, thereby maintaining normal bone metabolic balance. The role of vitamin D in bones is not fully understood, but currently known functions include: ① promoting bone matrix formation and osteoid mineralization; ② promoting osteoblast differentiation and the synthesis of proteins such as alkaline phosphatase and osteocalcin; ③ inducing the expression of nuclear factor-κB receptor activator ligands, thereby promoting osteoblast proliferation and osteoclast activation. Therefore, vitamin D has a direct stimulatory effect on osteoblasts and an indirect stimulatory effect on osteoclasts.

[0003] Vitamin D deficiency is a series of symptoms resulting from insufficient vitamin D. Low serum calcium levels due to vitamin D deficiency trigger parathyroid hormone to stimulate osteoclasts to resorb bone, releasing minerals stored in the bones into the bloodstream to maintain normal blood calcium levels. If parathyroid hormone levels remain elevated, causing a decrease in serum phosphate, bone diseases (rickets and osteomalacia) can occur. The incidence of vitamin D deficiency is high, approximately 30% to 50%, requiring supplementation. Rickets now occurs not only in temperate regions with limited sunlight but also in sunny areas. Vitamin D is fat-soluble; to address its solubility, traditional methods typically prepare it as oil-based drops. Vitamin D is extremely sensitive to heat, light, and oxygen, and is easily degraded. Formulations containing large amounts of antioxidants and oily solvents pose safety risks, especially to high-risk groups for vitamin D deficiency (infants, pregnant women, and the elderly). The largest group of people with vitamin D deficiency are newborns and infants. According to the "Dietary Guidelines for Chinese Residents", the recommended dosage of vitamin D supplementation should be adjusted according to body weight. Single-dose drops (capsules) cannot accurately adjust the dosage according to needs and cannot fully meet clinical requirements. Summary of the Invention

[0004] Based on this, the purpose of this application includes providing a vitamin D nanomicelle formulation and its preparation method, wherein the vitamin D nanomicelle formulation does not contain antioxidants, has good stability, high safety, adjustable dosage, and high compliance.

[0005] To achieve the above objectives, the following technical solution is provided:

[0006] A vitamin D nanomicelle formulation comprising vitamin D as the main ingredient and excipients;

[0007] The excipients include polyethylene glycol 15-hydroxystearate and glycocholic acid;

[0008] The weight ratio of the vitamin D, the 15-hydroxystearic acid polyethylene glycol ester, and the glycocholic acid is 1:(5~20):(2~10).

[0009] In one embodiment, the excipient further includes citric acid;

[0010] The citric acid accounts for 0.05% to 0.5% of the total weight of the vitamin D nanomicelle formulation.

[0011] A vitamin D nanomicelle formulation, wherein the vitamin D nanomicelle formulation is a solution, and the pH value of the solution is 4.5-6.5;

[0012] Each 1 mL of the solution comprises: 400 IU to 800 IU of vitamin D, 100 mcg to 400 mcg of 15-hydroxystearic acid polyethylene glycol ester, 40 mcg to 200 mcg of glycocholic acid, and a solvent.

[0013] In one embodiment, each 1 mL of the solution further includes 0.05% to 0.5% citric acid.

[0014] A method for preparing a vitamin D nanomicelle formulation includes the following steps:

[0015] 15-hydroxystearic acid polyethylene glycol ester is heated to 45℃~55℃, and while maintaining the temperature at 45℃~55℃, glycocholic acid is added and mixed, followed by vitamin D and mixed to obtain a first mixture. The weight ratio of vitamin D, 15-hydroxystearic acid polyethylene glycol ester and glycocholic acid is 1:(10~30):(4~12).

[0016] The solvent is heated to 45℃~55℃ and then added to the first mixture to obtain the second mixture;

[0017] Optionally, citric acid is added to the second mixture to prepare a third mixture;

[0018] The pH of the second mixture or the third mixture is adjusted to 4.5-6.5, filtered, and the vitamin D nanomicelle preparation is obtained.

[0019] In one embodiment, the stirring speed for adding glycocholic acid is 150 r / min to 300 r / min; and / or

[0020] The stirring speed for adding glycocholic acid is 150 r / min to 300 r / min; and / or

[0021] The solvent is heated to 45℃~55℃ and then added to the first mixture. The stirring speed is 150r / min~300r / min.

[0022] In one embodiment, the second mixture is cooled to 20°C to 30°C before adding citric acid to the mixture.

[0023] In one embodiment, the pH value of the second mixture or the third mixture is adjusted using an aqueous sodium hydroxide solution.

[0024] In one embodiment, the light intensity during the preparation of the vitamin D nanomicelle formulation does not exceed 200 LX.

[0025] In one embodiment, filtration is performed using a filter membrane with a pore size of 0.2 μm to 0.25 μm.

[0026] This application describes the self-assembly of fat-soluble vitamin D with polyethylene glycol 15-hydroxystearic acid and glycocholic acid to form nanomicelles. The vitamin D nanomicelles have a uniform particle size distribution, which improves the dispersibility of vitamin D in water. The particle size can be kept stable within a certain range, and they are less likely to aggregate during long-term storage, which is more conducive to the stability of the drug.

[0027] The preparation method described in this application is simple to operate, uses readily available raw materials, and is low in cost, making it suitable for industrial production and application. Attached Figure Description

[0028] To more clearly illustrate the technical solutions in the specific embodiments of this application or the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0029] Figure 1 The particle size detection results are for the vitamin D nanomicelles prepared in Example 1 of this application;

[0030] Figure 2 The particle size distribution of vitamin D nanomicelles prepared in Comparative Example 1 of this application is shown. Detailed Implementation

[0031] The present application is further described below with reference to embodiments and examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the application. Furthermore, it should be understood that after reading the teachings of this application, those skilled in the art can make various alterations or modifications to this application, and these equivalent forms also fall within the protection scope of the appended claims.

[0032] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0033] the term

[0034] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0035] The terms "and / or," "or / and," and "and / or" as used herein include any one of two or more related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations encompass any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that, in this application, the technical solution undoubtedly includes solutions connected by "logical AND," and also undoubtedly includes solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0036] In this document, terms such as "preferred," "better," and "more preferred" are merely descriptions of implementation methods or examples that achieve better results, and should be understood as not constituting a limitation on the scope of protection of this application.

[0037] In this application, terms such as "further," "even further," and "particularly" are used to describe purposes and indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0038] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0039] In this application, numerical intervals (i.e., numerical ranges) are involved. Unless otherwise specified, optional numerical distributions within the aforementioned numerical intervals are considered continuous and include the two endpoints (i.e., the minimum and maximum values) of the numerical range, as well as every value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints. Furthermore, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0040] In this application, weight can be a well-known unit of mass in the chemical industry, such as μg, mg, g, or kg. The weight unit "mcg" refers to micrograms, which is equivalent to "μg".

[0041] In this application, IU is a medical potency unit, commonly used for dosage of fat-soluble vitamins. A possible conversion method between IU and weight is: 1 IU vitamin D = 0.025 μg vitamin D3.

[0042] In this application, unless otherwise specified, the terms "size", "particle size", and "diameter" generally refer to average values.

[0043] Vitamin D is an essential trace element for maintaining physiological functions. Besides dietary intake, some people require additional vitamin D supplements. Vitamin D is a fat-soluble vitamin with a hydrophobic molecular structure, making it unsuitable for salt formation and exhibiting poor stability. Surfactants are commonly used to solubilize vitamin D in aqueous oral solutions. Surfactants can be ionic or nonionic. Because ionic surfactants are more toxic, nonionic surfactants, such as sorbitan derivatives and polyoxyethylene fatty acid esters, are generally used in pharmaceutical formulations. Using only a single surfactant as a co-solvent requires a large quantity, often results in opalescence, and fails to address the poor stability of vitamin D. This application, through extensive research, has discovered that nonionic surfactants, under specific conditions, combine with appropriate glycocholic acid to form micelles with small particle size, good external hydrophilicity, and a large internal hydrophobic core. These micelles can encapsulate vitamin D within the internal hydrophobic core, increasing its solubility and, more importantly, significantly improving its stability.

[0044] One aspect of this application provides a vitamin D nanomicelle formulation comprising vitamin D as the main ingredient and excipients;

[0045] The excipients include polyethylene glycol 15-hydroxystearate and glycocholic acid;

[0046] The weight ratio of vitamin D, 15-hydroxystearic acid polyethylene glycol ester and glycocholic acid is 1:(5~20):(2~10).

[0047] Glycinecholic acid and polyethylene glycol 15-hydroxystearate (PEG) form micelles. Glycinecholic acid can be embedded within PEG molecules, resulting in a micelle solution with small particle size and a clear, transparent appearance. Because the number of glycocholic acid molecules that can be embedded within PEG molecules is limited, there is a limit to the ratio of glycocholic acid to PEG. If the mass ratio of PEG to PEG exceeds the maximum mass ratio, the number of PEG molecules increases, making it difficult to form mixed micelles. Some PEG molecules tend to aggregate, forming larger aggregated micelles. The resulting solution exhibits a multi-peaked, uneven particle size distribution, and contains large aggregated micelles.

[0048] In one embodiment, the weight ratio of 15-hydroxystearic acid polyethylene glycol ester to glycocholic acid is 100:(25~50), preferably 100:(30~40).

[0049] In one embodiment, the weight ratio of vitamin D, the 15-hydroxystearic acid polyethylene glycol ester, and the glycocholic acid is 1:(10~20):(5~10).

[0050] In one embodiment, the excipients also include citric acid;

[0051] Citric acid accounts for 0.05% to 0.5% of the total weight of vitamin D nanomicelle formulations.

[0052] In another aspect of this application, a vitamin D nanomicelle formulation is provided. This vitamin D nanomicelle formulation is a solution. The vitamin D micelles exhibit aggregation under high-temperature instability in the solvent dispersion medium, and the particle size distribution shows multiple peaks. By adding an appropriate amount of citric acid and adjusting the pH to different values ​​(4.5–6.5), the particle size distribution not only exhibits a single peak during the stability period and has a low PDI value (<0.3), but the particle size value can also remain stable within a certain range.

[0053] In one embodiment, each 1 mL of vitamin D nanomicelle formulation comprises: 800 IU of vitamin D, 100 mcg to 400 mcg of polyethylene glycol 15-hydroxystearate, 40 mcg to 200 mcg of glycocholic acid, and a solvent.

[0054] In one embodiment, each 1 mL of vitamin D nanomicelle formulation comprises: 800 IU of vitamin D, 300 mcg to 400 mcg of polyethylene glycol 15-hydroxystearate, 10 mcg to 200 mcg of glycocholic acid, and a solvent.

[0055] In one embodiment, the vitamin D nanomicelle formulation is a solution with a pH value of 4.5 to 6.5;

[0056] Each 1 mL of the solution comprises: 800 IU of vitamin D, 100 mcg to 400 mcg of polyethylene 15-hydroxystearic acid, 40 mcg to 200 mcg of glycocholic acid, and a solvent. In one embodiment, the solvent is water.

[0057] In one embodiment, citric acid accounts for 0.05% to 0.5% of the total weight of the vitamin D nanomicelle formulation in the solution.

[0058] In another aspect of this application, a method for preparing a vitamin D nanomicelle formulation is provided, comprising the following steps:

[0059] 15-hydroxystearic acid polyethylene glycol ester is heated to 45℃~55℃, and while maintaining the temperature at 45℃~55℃, glycocholic acid is added and mixed, followed by vitamin D and mixed to obtain the first mixture. The weight ratio of vitamin D, 15-hydroxystearic acid polyethylene glycol ester and glycocholic acid is 1:(5~20):(2~10).

[0060] The solvent is heated to 45℃~55℃ and then added to the first mixture to obtain the second mixture;

[0061] Optionally, citric acid is added to the second mixture to prepare a third mixture;

[0062] Adjust the pH of the second or third mixture to 4.5-6.5, filter, and obtain a vitamin D nanomicelle preparation.

[0063] In one embodiment, the stirring speed for adding glycocholic acid is 150 r / min to 300 r / min; and / or

[0064] The stirring speed for adding glycocholic acid is 150 r / min to 300 r / min; and / or

[0065] The solvent is heated to 45℃~55℃ and then added to the first mixture. The stirring speed is 150r / min~300r / min.

[0066] In one embodiment, the second mixture is cooled to 20°C to 30°C before adding citric acid to the mixture.

[0067] In one embodiment, the pH of the second or third mixture is adjusted using an aqueous sodium hydroxide solution.

[0068] In one embodiment, the light intensity during the preparation of the vitamin D nanomicelle formulation does not exceed 200 LX.

[0069] In one embodiment, filtration is performed using a filter membrane with a pore size of 0.2 μm to 0.25 μm.

[0070] The following are some specific examples.

[0071] For experimental parameters not specified in the following specific embodiments, please refer to the guidelines given in this application document first, or refer to experimental manuals or other experimental methods known in the art, or refer to the experimental conditions recommended by the manufacturer.

[0072] The raw materials and reagents involved in the following specific embodiments can be obtained commercially or prepared by those skilled in the art using known methods.

[0073] The vitamin D used in the following examples is vitamin D3, pure powder; manufacturer: Sichuan Yuxin Pharmaceutical Co., Ltd.; batch number: 210601, 220403.

[0074] 1. Preparation of Vitamin D Nanomicelle Solution

[0075] 1.1 Preparation of Vitamin D Nanomicelle Solution according to the Formulation in Table 1 Example

[0076] (1) Ingredients

[0077] Prepare the raw materials for preparing vitamin D nanomicelle solution according to Table 1.

[0078] Table 1

[0079]

[0080] In Table 1, "-" indicates that the item was not added in this embodiment.

[0081] (2) Prepare vitamin D nanomicelle solution according to the following steps. The entire preparation process should be carried out in the dark (the light intensity should not exceed 200 LX).

[0082] 1) Heat 15-hydroxystearic acid polyethylene glycol ester to 45℃~55℃ to melt it, and add glycocholic acid while maintaining the temperature at 45℃~55℃ and stir until completely dissolved. The stirring speed is 150r / min~300r / min.

[0083] 2) Add vitamin D to step (1), and add the prescribed amount of glycinecholic acid while maintaining a temperature of 45℃~55℃ and stirring until completely dissolved. The stirring speed is 150r / min~300r / min.

[0084] 3) Add purified water / water for injection at 45℃~55℃ to the solution in step (2), and stir for 10-20 minutes at a stirring speed of 150r / min~300r / min;

[0085] 4) Cool the solution from step 3 to below 30°C, add citric acid, sucralose, and strawberry flavoring, and stir to dissolve;

[0086] 5) Adjust the pH value with sodium hydroxide solution, filter at 0.22μm, and then fill into containers.

[0087] 1.2 Prepare comparative vitamin D nanomicelle solutions according to the formulations in Tables 2 and 3.

[0088] Prepare the comparative vitamin D nanomicelle solution according to ingredients 2-3, using essentially the same method as the preparation example of the vitamin D nanomicelle solution, including formulations 1-10.

[0089] Table 2

[0090]

[0091] In Table 2, "-" indicates that the item was not added in the comparative example.

[0092] Table 3

[0093]

[0094] In Table 3, "-" indicates that the item was not added in the comparative example.

[0095] 2. Performance Testing

[0096] (1) Particle size test

[0097] The particle size and particle size distribution of the samples in each example and comparative example were detected by dynamic light scattering method (nanoparticle size and molecular weight analyzer, Malvern Zetasizernano ZS). The results are shown in Tables 4, 5 and 6.

[0098] Table 4. Detection results of particle size and particle size distribution in Examples 1-3

[0099]

[0100] Table 5. Detection results of particle size and particle size distribution of formulations 1-5 in the comparative examples.

[0101]

[0102] Table 6. Detection results of particle size and particle size distribution of formulations 6-10 in the comparative examples.

[0103]

[0104] (2) Stability test

[0105] High-performance liquid chromatography (HPLC) was used to detect the content and related substances. The chromatographic conditions included:

[0106] Chromatographic column: Inertsil ODS-2 column (150×4.6mm, 5μm);

[0107] Flow rate: 1.5 ml / min;

[0108] Detection wavelength: 265nm;

[0109] Detector: Ultraviolet detector;

[0110] Mobile phase: Methanol: Acetonitrile = 350:650;

[0111] The vitamin D nanomicelle liquid formulations of each formulation in Examples 1 and 3 were subjected to accelerated and long-term stability tests in accordance with the guidelines for drug stability testing in the appendix of the 2020 edition of the Chinese Pharmacopoeia. The results are shown in Table 7. According to Table 7, it can be found that the vitamin D nanomicelle liquid formulations of each example meet the quality standard requirements.

[0112] Table 7. Stability test results of the vitamin D nanomicelle liquid formulation in the examples.

[0113]

[0114] All references to this application are incorporated herein by reference as if each document were individually incorporated herein by reference. Unless they conflict with the purpose and / or technical solution of this application, all cited references are incorporated herein by reference in their entirety and for all purposes. When references are cited in this application, the definitions of relevant technical features, terms, nouns, phrases, etc., are also incorporated herein by reference. Examples and preferred embodiments of the cited technical features may also be incorporated herein by reference, but only to the extent that they enable the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, this application shall prevail or modifications shall be made adaptably to the description in this application.

[0115] The technical features of the above-described embodiments and examples can be combined in any suitable manner. For the sake of brevity, not all possible combinations of the technical features in the above-described embodiments and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0116] The embodiments described above merely illustrate several implementation methods of this application and should not be construed as limiting the scope of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Furthermore, it should be understood that after reading the above teachings, those skilled in the art can make various alterations or modifications to this application, and the equivalent forms obtained also fall within the protection scope of this application. It should also be understood that technical solutions obtained by those skilled in the art based on the technical solutions provided in this application through logical analysis, reasoning, or limited experimentation are all within the protection scope of the appended claims. Therefore, the protection scope of this patent application should be determined by the appended claims, and the specification can be used to interpret the content of the claims.

Claims

1. A vitamin D nanomicelle formulation, characterized in that, It includes the main ingredient, vitamin D, and auxiliary ingredients; The excipients include polyethylene glycol 15-hydroxystearic acid, glycocholic acid, citric acid, and purified water; The weight ratio of the vitamin D, the 15-hydroxystearic acid polyethylene glycol ester, and the glycocholic acid is 1:(10~20):(5~10). The citric acid accounts for 0.05% to 0.5% of the total weight of the vitamin D nanomicelle formulation; The vitamin D nanomicelle formulation is an aqueous solution with a pH value of 4.5 to 6.

5.

2. A method for preparing a vitamin D nanomicelle formulation, characterized in that, Includes the following steps: 15-hydroxystearic acid polyethylene glycol ester is heated to 45℃~55℃, and while maintaining the temperature at 45℃~55℃, glycocholic acid is added and mixed, followed by vitamin D to obtain a first mixture. The weight ratio of vitamin D, 15-hydroxystearic acid polyethylene glycol ester, and glycocholic acid is 1:(10~20):(5~10). The purified water was heated to 45℃~55℃ and then added to the first mixture to obtain the second mixture. Citric acid was added to the second mixture to prepare a third mixture; the citric acid accounted for 0.05% to 0.5% of the total weight of the vitamin D nanomicelle formulation. The pH of the third mixture was adjusted to 4.5-6.5, and the mixture was filtered to obtain the vitamin D nanomicelle preparation.

3. The method for preparing the vitamin D nanomicelle formulation as described in claim 2, characterized in that, The stirring speed for adding glycocholic acid is 150 r / min to 300 r / min; and / or The stirring speed for adding glycocholic acid is 150 r / min to 300 r / min; and / or The purified water was heated to 45℃~55℃ and then added to the first mixture. The stirring speed was 150r / min~300r / min.

4. The method for preparing the vitamin D nanomicelle formulation as described in claim 2, characterized in that, Before adding citric acid to the second mixture, cool the second mixture to 20°C~30°C.

5. The method for preparing the vitamin D nanomicelle formulation as described in claim 2, characterized in that, The pH of the second or third mixture is adjusted using an aqueous sodium hydroxide solution.

6. The method for preparing the vitamin D nanomicelle formulation according to any one of claims 2-5, characterized in that, The light intensity during the preparation of the vitamin D nanomicelle formulation is no higher than 200 LX.

7. The method for preparing the vitamin D nanomicelle formulation according to any one of claims 2-5, characterized in that, Filter using a filter membrane with a pore size of 0.2μm to 0.25μm.

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