A liposome composition, its preparation method and use thereof

Abstract

The application discloses a liposome composition and a preparation method and application thereof, and belongs to the technical field of liposome solution preparation. The liposome composition comprises liposomes and a compound emulsifier, the liposomes contain bioactive ingredients, the compound emulsifier is composed of sorbitan peristerate and sorbitan sesquioleate with a mass ratio of (1:3) to (3:1), and the liposome composition does not contain an aqueous phase. The liposome composition can dissolve water-soluble bioactive ingredients in an oily base, ensures that the system is clear, and does not change the molecular structure of the bioactive ingredients, thereby retaining the bioactivity of the bioactive ingredients to the maximum extent.

Description

Technical Field

[0001] This invention relates to the field of liposome solution preparation technology, and in particular to a liposome composition, its preparation method, and its application. Background Technology

[0002] Nicotinamide riboside (NR) is a naturally occurring vitamin B3 derivative, belonging to the nucleoside class of compounds. In vivo, NR is converted to nicotinamide mononucleotide (NMN) by nicotinamide ribokinase, and NMN is then catalyzed by NMN adenylate transferase to produce nicotinamide adenine dinucleotide (NAD). + As NAD + Nicotinamide riboside, a direct precursor of NAD+, can be efficiently converted into NAD+ in vivo. + This supports cellular energy metabolism, DNA repair, and the activation of longevity proteins. Studies have shown that nicotinamide ribose can directly inhibit the activity of enzymes related to melanin synthesis, thereby improving hyperpigmentation of the skin; at the same time, nicotinamide ribose can exert a protective effect on the skin layer by inhibiting the expression of degradation enzymes in damaged skin, showing great potential in the fields of medical aesthetics and skin care.

[0003] In recent years, with the growing acceptance and explosive application of the "oil-based skincare" concept, oil-based skincare products such as facial oils and repair oils have become a noteworthy category in the beauty industry. However, currently available niacinamide ribose raw materials are all water-soluble and cannot exist stably in oil-based formulations, easily precipitating out and causing the system to become cloudy. Furthermore, the clarity of oil-soluble raw materials is crucial for the clear and transparent final product; only clear and transparent oil-soluble raw materials can be used in transparent final systems. Cloudy, flocculent, or solidified oil-soluble raw materials will reduce the transparency and stability of the finished product.

[0004] Therefore, to meet the efficacy requirements of oil-based skincare products, it is necessary to develop oil-soluble niacinamide ribose raw materials to solve the problem of its incompatibility in oil-based formulation systems. NR is unstable in aqueous solutions, difficult to apply in powder form, and inconvenient to weigh. Therefore, there is an urgent need for a clear and stable niacinamide ribose composition with an oil-phase matrix. Summary of the Invention

[0005] To address the current lack of nicotinamide-ribose compositions with an oil phase matrix, this invention provides a lipid composition containing water-soluble bioactive components that can be directly added to the oil phase. This not only ensures the clarity of the oil phase system but also preserves the molecular structure of the bioactive components, maximizing their bioactivity. This invention is achieved through the following scheme: A liposome composition comprising liposomes and a compound emulsifier, the liposomes containing a bioactive ingredient, the compound emulsifier comprising sorbitan polyether-30 tetraisostearate and sorbitan sesquioleate in a mass ratio of (1:3) to (3:1), the liposome composition being free of an external aqueous phase.

[0006] In some embodiments, the mass ratio of sorbitan polyether-30 tetraisostearate to sorbitan sesquioleate is (1:2) to (2:1), preferably (1:1.5) to (1.5:1), for example, 1:1.

[0007] In some embodiments, the concentration of the liposomes in the emulsifier is 0.1wt%-30wt%; preferably 0.5wt%-20wt%, more preferably 5wt%-20wt%, and even more preferably 8wt%-15wt%, for example 10wt%.

[0008] In some embodiments, the bioactive ingredient is a water-soluble ingredient, preferably nicotinamide ribose and its salts, such as nicotinamide ribose, nicotinamide ribose fumarate, nicotinamide ribose malate, and / or nicotinamide ribose chloride.

[0009] In some embodiments, the liposomes are prepared by the following steps: Mixing: The bioactive component and lipid component are mixed with water and a stabilizer to obtain a mixed solution; Dispersion: The above mixed solution is emulsified and dispersed to obtain a dispersion; Homogenization: The above dispersion was homogenized under high pressure to obtain a liposome suspension; Freeze-drying: The external aqueous phase in the above liposome suspension is removed by freeze-drying to obtain solid liposomes.

[0010] In some embodiments, the stabilizer in the mixing step is selected from propylene glycol, for example 1,2-propanediol and / or 1,3-propanediol.

[0011] In some embodiments, during the mixing step, the lipid component is selected from lecithin and its derivatives, preferably at least one of lecithin, hydrogenated lecithin and distearate, more preferably hydrogenated lecithin.

[0012] In some embodiments, during the mixing step, the phosphatidylcholine content in the lipid component is 20 wt%-90 wt%, preferably 30 wt%-80 wt%, more preferably 45 wt%-75 wt%, and even more preferably 60 wt%-75 wt%, for example 70 wt%.

[0013] In some embodiments, during the mixing step, the bioactive ingredient has a mass percentage concentration of 0.05%-0.3 wt% in the mixed solution; preferably 0.08%-0.25 wt%, for example 0.1 wt%.

[0014] In some embodiments, during the mixing step, the mass percentage concentration of the stabilizer in the mixed solution is 5%-20 wt%, preferably 8%-15 wt%, more preferably 8%-12 wt%, for example 10 wt%.

[0015] In some embodiments, during the mixing step, the mass percentage concentration of the lipid component in the mixed solution is 0.05 wt%-2 wt%, preferably 0.1 wt%-1.5 wt%, more preferably 0.3 wt%-1 wt%, for example 0.5 wt%.

[0016] In some embodiments, the emulsification dispersion in the dispersion step is performed using an emulsification disperser.

[0017] In some embodiments, during the dispersion step, the rotation speed of the emulsification dispersion is 1000 rpm / min-5000 rpm / min, preferably 2000 rpm / min-4000 rpm / min, for example 2500 rpm / min.

[0018] In some embodiments, the emulsification dispersion time in the dispersion step is 5 min-30 min, preferably 10 min-20 min, for example 15 min.

[0019] In some embodiments, the homogenization step involves pre-cooling the dispersion, preferably to 3-5°C.

[0020] In some embodiments, the homogenization step involves 2-10 cycles of homogenization, preferably 3-8 cycles, for example, 5 cycles.

[0021] In some embodiments, the homogenization step involves a homogenization pressure of 700-1000 bar during high-pressure homogenization.

[0022] In some of these embodiments, the freeze-drying step is performed using a gradient freezing process.

[0023] In some implementations, the gradient freezing procedure is as follows: Pre-freezing stage: temperature -40±5℃, freezing time 2.0-6h, atmospheric pressure; Primary drying stage: temperature -20±3℃, freezing time 10±3h; pressure 0.01±0.001mBar; Secondary drying stage: temperature -10±1℃, freezing time 10±1h; pressure 0.01±0.001mBar.

[0024] In some implementations, the gradient freezing procedure is as follows: Pre-freezing stage: temperature -40℃, freezing time 2.0-6h, atmospheric pressure; Primary drying stage: temperature -20℃, freezing time 10h; pressure 0.01mBar; Secondary drying stage: temperature -10℃, freezing time 10h; pressure 0.01mBar.

[0025] On the other hand, the present invention discloses a method for preparing the above-mentioned liposome composition, comprising the following steps: the liposomes are mixed with the compound emulsifier at a temperature of 60°C-80°C to obtain the liposomes; preferably, the mixing temperature is 65°C-75°C, for example, 70°C.

[0026] On the other hand, the present invention also discloses a liposome oil solution comprising the above-described liposome composition and an oily matrix.

[0027] In some embodiments, the oily matrix is ​​selected from at least one of: caprylic / capric triglyceride, squalane, dioctyl carbonate, isononyl isononanoate, isohexadecane, C15-19 alkyl, sunflower seed oil, olive oil, and jojoba oil, preferably at least one of dioctyl carbonate, isononyl isononanoate, isohexadecane, C15-19 alkyl, sunflower seed oil, olive oil, and jojoba oil.

[0028] In some embodiments, the liposome composition comprises 1 wt%-30 wt% of the oily matrix, preferably 5 wt%-20 wt%, and more preferably 15 wt%-20 wt%.

[0029] In some embodiments, the oily matrix is ​​selected from at least one of caprylic / capric triglycerides and squalane, and the liposome composition accounts for 15wt%-20wt% of the oily matrix by mass.

[0030] In some embodiments, the oily matrix is ​​selected from at least one of dioctyl carbonate, isononyl isononanoate, isohexadecane, C15-19 alkyl, sunflower seed oil, olive oil, and jojoba oil, and the liposome composition accounts for 5 wt% to 20 wt% of the oily matrix by mass.

[0031] On the other hand, the present invention also discloses a method for preparing the above-mentioned liposome oil solution, comprising the following steps: the liposome composition and the oily matrix are mixed evenly to obtain the solution.

[0032] In some embodiments, the liposome composition is incorporated into the oily matrix.

[0033] On the other hand, the present invention also discloses the use of the above-mentioned liposome composition and the above-mentioned liposome oil solution in preparing water-soluble bioactive ingredients into oil solutions.

[0034] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0035] definition: "External aqueous phase" refers to the independent phase region in a liposome dispersion system or multiphase composition in which water or an aqueous solution (such as buffer solution, physiological saline, pure water, etc.) serves as the continuous phase (external phase) or dispersion medium, which is distinct from the "internal aqueous phase" (an aqueous core enclosed by a lipid bilayer) that may exist in liposomes.

[0036] "Water-soluble components" refer to substances that have a higher solubility in aqueous solutions than in oily matrices. Common water-soluble bioactive components that serve as the in vivo phase of lipids include small molecule drugs, nucleic acid drugs, vaccine antigens, contrast agents and diagnostic markers, and vitamins and nutrients.

[0037] "Oily matrix" refers to the hydrophobic substance used to disperse liposomes, which is liquid or semi-solid at room temperature or can be converted into a liquid state by heating. Common oily matrices include natural fats and oils, mineral oils, synthetic or semi-synthetic esters, fatty acids and their derivatives, silicone oils, and wax mixtures.

[0038] The reagents and raw materials used in this invention are all commercially available.

[0039] The positive and progressive effects of this invention are as follows: This invention combines liposome encapsulation technology with emulsification to prepare an oil-soluble NR liposome composition, which dissolves water-soluble NR in an oily matrix, ensuring a clear system; at the same time, it does not change the molecular structure of NR, thus preserving the biological activity of NR to the greatest extent. Attached Figure Description

[0040] Figure 1 Photographs of liposome compositions with different formulations of emulsifiers.

[0041] Figure 2 Photographs showing the solubility of liposome compositions of different concentrations in the caprylic / capric triglyceride system.

[0042] Figure 3Photographs showing the solubility of liposome compositions of different concentrations in a squalane system.

[0043] Figure 4 Photographs showing the solubility of liposome compositions of different concentrations in a dioctyl carbonate system.

[0044] Figure 5 Photographs showing the solubility of liposome compositions of different concentrations in sunflower seed oil system. Detailed Implementation

[0045] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0046] Reagent source: Sorbitol polyether-30 tetraisostearate (CAS: 64364-10-9): Manufacturer: Jiangxi Shouxin New Materials Co., Ltd., Model: SX-STI430.

[0047] Sorbitan sesquioleate (CAS: 8007-43-0): Manufacturer: Croda Chemicals (Shanghai) Co., Ltd., Model: SP SPAN 83 MBAL-LQ-(SG).

[0048] PEG-20 glyceryl triisostearate (CAS: 86846-21-1): Manufacturer: Miyoshi Oils & Fats Co., Ltd., Japan; Model: M FINEOIL ISG-20T.

[0049] Sorbitan isostearate (CAS: 71902-01-7): Manufacturer: OLEON NV, Model: Radiasurf 7758.

[0050] Example

[0051] Liposome oil solutions were prepared using the following method: I. Preparation Method 1. Liposome preparation The above raw materials were added to deionized water with a concentration of 0.1 wt% nicotinamide riboflavin, 10 wt% 1,3-propanediol, and 0.5 wt% hydrogenated lecithin to obtain mixed solution A; wherein the phosphatidylcholine content of hydrogenated lecithin was 70 wt%.

[0052] The above solution A was used to disperse the raw materials evenly using a high-speed emulsifying disperser at a speed of 2500 rpm / min for 15 min to obtain dispersion B.

[0053] The above dispersion B was pre-cooled to 3-5℃ and circulated 5 times using a high-pressure homogenizer at a homogenization pressure of 700-1000 bar to obtain liposome suspension C.

[0054] 2. Preparation of solid liposomes

[0055] The above liposome suspension C was subjected to vacuum freeze-drying using the following process: Pre-freezing stage: temperature -40℃, freezing time 2.0-6h, atmospheric pressure; Primary drying stage: temperature -20℃, freezing time 10h; pressure 0.01mBar; Secondary drying stage: temperature -10℃, freezing time 10h; pressure 0.01mBar; After removing all the water, solid liposomes are obtained.

[0056] 3. Preparation of liposome compositions

[0057] The above-mentioned liposome solids were added to emulsifiers with different formulations at an addition amount of 10 wt%, and dissolved at 70°C to obtain different liposome compositions.

[0058] Table 1. Different emulsifier formulations for 10 wt% liposome compositions

[0059] Note: " / " indicates that it has not been added.

[0060] The above-mentioned liposome compositions composed of emulsifiers with different formulations are shown in the following figures. Figure 1 As shown in the figure, the liposome composition is clear when the mass ratio of sorbitan polyether-30 tetraisostearate to sorbitan sesquioleate is 1:1, 1:2, and 2:1.

[0061] 4. Preparation of liposome oil solution

[0062] To verify the solubility of the oil-soluble liposome composition in different oily matrices, the liposome composition obtained from Formulation 5 was added to different oily matrices at amounts of 5 wt%, 10 wt%, 15 wt%, and 20 wt%, respectively. The oily matrices included caprylic / capric triglycerides, squalane, dioctyl carbonate, isononyl isononanoate, isohexadecane, and C... 15-19 Alkane, silicone oil, sunflower seed oil, olive oil and jojoba oil (as shown in the table below) are mixed evenly to obtain a liposome oil solution containing nicotinamide ribose (NR).

[0063] The clarity of the mixed oil solution was observed and recorded. The results are shown in the table below. Figure 2-5 .

[0064] Table 2. Clarity of oil solutions with different oily matrices

[0065] Figure 2-5 The images show the liposome composition in representative oil phase matrices: caprylic / capric triglyceride, squalane, dioctyl carbonate, and sunflower seed oil. As can be seen from the tables and figures above, dioctyl carbonate, sunflower seed oil, isononyl isononanoate, isohexadecane, and C... 15-19 Alkane, olive oil, and jojoba oil remained clear throughout the entire concentration range from 5 wt% to 20 wt%, demonstrating excellent compatibility. Caprylic / capric triglycerides and squalane showed a concentration-dependent effect, with the solution becoming turbid at 5 wt% and 10 wt%, but becoming clear when the concentration increased to 15 wt% and above. Silicone oil, on the other hand, separated into layers at all tested concentrations, indicating poor compatibility.

Claims

1. A liposome composition, characterized in that, The composition includes liposomes and a compound emulsifier, wherein the liposomes contain bioactive ingredients, and the compound emulsifier comprises sorbitan polyether-30 tetraisostearate and sorbitan sesquioleate in a mass ratio of (1:3) to (3:1), and the liposome composition does not contain an external aqueous phase.

2. The liposome composition according to claim 1, characterized in that, The mass ratio of sorbitan polyether-30 tetraisostearate to sorbitan sesquioleate is (1:2) to (2:1), preferably (1:1.5) to (1.5:1), for example, 1:

1.

3. The liposome composition according to claim 1, characterized in that, Meets at least one of the following conditions: (1) The concentration of the liposomes in the emulsifier is 0.1wt%-30wt%; preferably 0.5wt%-20wt%, more preferably 5wt%-20wt%, and even more preferably 8wt%-15wt%, for example 10wt%; (2) The bioactive ingredient is a water-soluble ingredient, preferably nicotinamide ribose and its salts, such as nicotinamide ribose, nicotinamide ribose fumarate, nicotinamide ribose malate, and / or nicotinamide ribose chloride.

4. The liposome composition according to claim 1, characterized in that, The liposomes were prepared by the following steps: Mixing: The bioactive component and lipid component are mixed with water and a stabilizer to obtain a mixed solution; Dispersion: The above mixed solution is emulsified and dispersed to obtain a dispersion; Homogenization: The above dispersion was homogenized under high pressure to obtain a liposome suspension; Freeze-drying: The external aqueous phase in the above liposome suspension is removed by freeze-drying to obtain solid liposomes.

5. The liposome composition according to claim 4, characterized in that, Meets at least one of the following conditions: (1) In the mixing step, the stabilizer is selected from: propylene glycol, for example: 1,2-propanediol and / or 1,3-propanediol; (2) In the mixing step, the lipid component is selected from: lecithin and its derivatives, preferably at least one of lecithin, hydrogenated lecithin and distearate phosphatidylcholine, more preferably hydrogenated lecithin; (3) In the mixing step, the content of phosphatidylcholine in the lipid component is 20 wt%-90 wt%, preferably 30 wt%-80 wt%, more preferably 45 wt%-75 wt%, and even more preferably 60 wt%-75 wt%, for example 70 wt%; (4) In the mixing step, the mass percentage concentration of the bioactive ingredient in the mixed solution is 0.05%-0.3wt%; preferably 0.08%-0.25wt%, for example 0.1wt%. (5) In the mixing step, the mass percentage concentration of the stabilizer in the mixed solution is 5%-20 wt%, preferably 8%-15 wt%, more preferably 8%-12 wt%, for example 10 wt%; (6) In the mixing step, the mass percentage concentration of the lipid component in the mixed solution is 0.05 wt%-2 wt%, preferably 0.1 wt%-1.5 wt%, more preferably 0.3 wt%-1 wt%, for example 0.5 wt%; (7) In the dispersion step, the emulsification dispersion is carried out using an emulsification disperser; (8) In the dispersion step, the rotation speed of the emulsification dispersion is 1000 rpm / min-5000 rpm / min, preferably 2000 rpm / min-4000 rpm / min, for example 2500 rpm / min; (9) In the dispersion step, the emulsification dispersion time is 5 min-30 min, preferably 10 min-20 min, for example 15 min; (10) In the homogenization step, the dispersion is first pre-cooled, preferably to 3-5°C; (11) In the homogenization step, the homogenization process is repeated 2-10 times, preferably 3-8 times, for example 5 times; (12) In the homogenization step, the homogenization pressure of the high-pressure homogenization is 700-1000 bar; (13) In the freeze-drying step, the freeze-drying is performed using a gradient freezing program; Preferably, the gradient freezing procedure is as follows: Pre-freezing stage: temperature -40±5℃, freezing time 2.0-6h, atmospheric pressure; Primary drying stage: temperature -20±3℃, freezing time 10±3h; pressure 0.01±0.001mBar; Secondary drying stage: temperature -10±1℃, freezing time 10±1h; pressure 0.01±0.001mBar; More preferably, the gradient freezing procedure is as follows: Pre-freezing stage: temperature -40℃, freezing time 2.0-6h, atmospheric pressure; Primary drying stage: temperature -20℃, freezing time 10h; pressure 0.01mBar; Secondary drying stage: temperature -10℃, freezing time 10h; pressure 0.01mBar.

6. A method for preparing the liposome composition according to any one of claims 1-5, characterized in that, Includes the following steps: The liposomes are mixed with the compound emulsifier at 60℃-80℃ to obtain the product; preferably, the mixing temperature is 65℃-75℃, for example, 70℃.

7. A liposome oil solution, characterized in that, It includes the liposome composition according to any one of claims 1-5 and the oily matrix.

8. The liposome oil solution according to claim 7, characterized in that, Meets at least one of the following conditions: (1) The oily matrix is ​​selected from at least one of the following: caprylic / capric triglyceride, squalane, dioctyl carbonate, isononyl isononanoate, isohexadecane, C15-19 alkyl, sunflower seed oil, olive oil and jojoba oil, preferably at least one of dioctyl carbonate, isonononyl isononanoate, isohexadecane, C15-19 alkyl, sunflower seed oil, olive oil and jojoba oil; (2) The liposome composition accounts for 1wt%-30wt% of the oily matrix, preferably 5wt%-20wt%, more preferably 15wt%-20wt%; (3) The oily matrix is ​​selected from at least one of caprylic / capric triglycerides and squalane, and the liposome composition accounts for 15wt%-20wt% of the oily matrix by mass. (4) The oily matrix is ​​selected from at least one of dioctyl carbonate, isononyl isononanoate, isohexadecane, C15-19 alkyl, sunflower seed oil, olive oil and jojoba oil, and the liposome composition accounts for 5wt%-20wt% of the oily matrix by mass.

9. The method for preparing the liposome oil solution according to any one of claims 7-8, characterized in that, Includes the following steps: The liposome composition and the oily matrix are mixed evenly to obtain the final product. Preferably, the liposome composition is added to the oily matrix.

10. Use of the liposome composition according to any one of claims 1-5 and the liposome oil solution according to any one of claims 7-8 in the preparation of a water-soluble bioactive ingredient into an oil solution.

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