Oil-soluble composition containing recombinant collagen and preparation method thereof

Abstract

The invention discloses an oil-soluble composition containing recombinant collagen. The composition comprises recombinant collagen, a polyhydroxy compound, grease, an emulsifier and water. The invention also provides a preparation method of the composition. The composition disclosed by the invention has the characteristics of good transparency, high drug loading capacity, good adaptability and good stability.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of cosmetics, and particularly relates to an oil-soluble composition containing recombinant collagen and a preparation method thereof. BACKGROUND

[0002] Collagen is the most abundant protein in mammals, accounting for 20-30% of the total protein mass of mammals, and widely exists in all tissues from the surface of lower vertebrates to the bodies of mammals. Collagen is also a very important protein in the human body and mainly exists in connective tissue. So far, about 28 different types of collagen have been found. The main reason why the skin of young people is smooth, full, and elastic is that there is relatively sufficient collagen. However, after the age of 20, the amount of collagen decreases by 10% per year, and the human body shows a series of symptoms such as wrinkles, sagging, long spots, dull skin, large pores, and sensitivity.

[0003] With the improvement of people's living quality and the pursuit of beauty, collagen products are widely used in medical and anti-aging and wrinkle-removing products. With the rapid development of gene recombination technology, recombinant human collagen expressed by Pichia pastoris and other microorganisms has high activity, low immunogenicity, low cost, and strong accessibility, and has significant advantages compared with animal collagen. It is rapidly popularized in the field of cosmetics. Recombinant collagen has good hydrophilicity and is also called soluble collagen, which is generally suitable for cosmetics with high water content, such as essence water, essence, and cream.

[0004] In recent years, products represented by essence oil for "nourishing skin with oil" have gradually emerged. The pure oil system has a unique skin feel, good moisturizing property, and better absorption of functional ingredients, and the proportion of skin care products has been increasing year by year. However, the oil solubility of recombinant collagen is poor, and it cannot be added to essence oil, makeup, and other products, which seriously limits the application of recombinant collagen in oil products.

[0005] There are solutions on the market that modify recombinant collagen to increase its oil solubility, but this method is high in cost, low in addition amount, and limited to small molecular collagen, and the prepared composition has poor stability and appearance, and is mostly turbid. CN 119074583 A discloses an oil-soluble wrapped anti-wrinkle and anti-aging composition. In this composition scheme, emulsifiers, oily solvents and other raw materials are used to wrap recombinant small molecular collagen, but the content is only 0.05-0.20%, the drug loading is very low, and it cannot meet the high content oil cosmetic addition demand. CN 118453442 A discloses an oil-soluble liquid composition containing recombinant collagen. The scheme mentions that the mass fraction of recombinant collagen powder in the composition is 5%-15%, but according to the preparation process of recombinant collagen powder recorded in the preparation examples 1-3 of the embodiment, "10% of trehalose corresponding to the weight of the recombinant collagen liquid, and 30% of mannitol corresponding to the weight of the recombinant collagen liquid are added into the recombinant type II collagen liquid, mixed, and then loaded into a freeze-drying tray and sent to a freeze-drying machine for freeze-drying treatment", it can be known that the main composition of the recombinant collagen powder obtained after freeze-drying is actually sugar, and the actual content of recombinant collagen is low. At the same time, a simple dispersion technology is used to disperse the recombinant collagen powder into oil, so the use range of the composition in this scheme is mainly limited to the addition of makeup.

[0006] Therefore, the present application provides an oil-soluble composition containing high drug loading of recombinant collagen, mainly solving the problem of oil solubility of recombinant collagen, and the oil-soluble composition prepared by using the composition has high drug loading, transparent appearance, good adaptability and good stability, and can be used for the addition of oil-based cosmetics such as essence oil, massage oil and makeup. SUMMARY

[0007] The present application relates to an oil-soluble composition of recombinant collagen, which has good transparency, high drug loading, good adaptability and good stability. The present application also provides a preparation method of the oil-soluble composition.

[0008] The first aspect of the present application relates to an oil-soluble composition, which comprises recombinant collagen, polyhydroxy compound, oil, emulsifier and water.

[0009] Among them, the mass percentage of recombinant collagen is 0.1%-5%; the mass percentage of polyhydroxy compound is 0.3%-15%; the mass percentage of oil is 30-90%; the mass percentage of emulsifier is 1-30%; and the mass percentage of water is 1%-20%;

[0010] The polyhydroxyl compound is selected from one or more of the group consisting of a bovine growth factor, glucose, maltose, trehalose, mannose, sucrose, maltose, fructose;

[0011] The oil is selected from one or more of the group consisting of an oil of plant origin, a synthetic oil, and a hydrocarbon-based oil.

[0012] The emulsifier is selected from polyethylene glycol (PEG) di-fatty acid ester and / or polyglyceryl di-fatty acid ester; the fatty acid is selected from one or more of the group consisting of stearic acid, palmitic acid, oleic acid, isostearic acid, polyhydroxystearic acid.

[0013] In some embodiments, the recombinant collagen has a mass percentage of 0.1%-5%. Preferably, the recombinant collagen has a mass percentage of 0.3%-5%. More preferably, the recombinant collagen has a mass percentage of 0.5%-5%. Even more preferably, the recombinant collagen has a mass percentage of 1%-5%. In some specific embodiments, the recombinant collagen has a mass percentage of 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, or 5%. In some specific embodiments, the recombinant collagen has a mass percentage of 0.1%. In some specific embodiments, the recombinant collagen has a mass percentage of 1%. In some specific embodiments, the recombinant collagen has a mass percentage of 2%. In some specific embodiments, the recombinant collagen has a mass percentage of 3%.

[0014] In some embodiments, the polyhydroxyl compound has a mass percentage of 0.3%-15%. Preferably, the polyhydroxyl compound has a mass percentage of 0.6%-15%. More preferably, the polyhydroxyl compound has a mass percentage of 1.5%-10%. Even more preferably, the polyhydroxyl compound has a mass percentage of 3%-10%. In some specific embodiments, the polyhydroxyl compound has a mass percentage of 0.6%, 1%, 1.5%, 2%, 3%, 5%, 8%, or 10%. In some specific embodiments, the polyhydroxyl compound has a mass percentage of 1%. In some specific embodiments, the polyhydroxyl compound has a mass percentage of 2%. In some specific embodiments, the polyhydroxyl compound has a mass percentage of 3%. In some specific embodiments, the polyhydroxyl compound has a mass percentage of 5%. In some specific embodiments, the polyhydroxyl compound has a mass percentage of 8%. In some specific embodiments, the polyhydroxyl compound has a mass percentage of 10%.

[0015] In some embodiments, the oil and fat is present in a mass percentage of 30-90%. Preferably, the oil and fat is present in a mass percentage of 40-85%. More preferably, the oil and fat is present in a mass percentage of 40-75%. In some specific embodiments, the oil and fat is present in a mass percentage of 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75%. In some specific embodiments, the oil and fat is present in a mass percentage of 40%. In some specific embodiments, the oil and fat is present in a mass percentage of 65%. In some specific embodiments, the oil and fat is present in a mass percentage of 70%. In some specific embodiments, the oil and fat is present in a mass percentage of 75%.

[0016] In some embodiments, the emulsifier is present in a mass percentage of 1-25%. Preferably, the emulsifier is present in a mass percentage of 1-20%. More preferably, the emulsifier is present in a mass percentage of 5-20%. In some specific embodiments, the emulsifier is present in a mass percentage of 5%, 10%, 15%, or 20%. In some specific embodiments, the emulsifier is present in a mass percentage of 5%. In some specific embodiments, the emulsifier is present in a mass percentage of 10%. In some specific embodiments, the emulsifier is present in a mass percentage of 15%. In some specific embodiments, the emulsifier is present in a mass percentage of 20%.

[0017] In some embodiments, the water is present in a mass percentage of 1-20%. Preferably, the water is present in a mass percentage of 3-20%. More preferably, the water is present in a mass percentage of 5-20%. In some specific embodiments, the water is present in a mass percentage of 5%, 10%, 15%, or 20%. In some specific embodiments, the water is present in a mass percentage of 5%. In some specific embodiments, the water is present in a mass percentage of 10%. In some specific embodiments, the water is present in a mass percentage of 15%. In some specific embodiments, the water is present in a mass percentage of 20%.

[0018] In some embodiments, the mass ratio of the recombinant collagen, the polyhydroxyl compound, and the water is 1:(1.5-10):(1-20); preferably, the mass ratio of the recombinant collagen, the polyhydroxyl compound, and the water is 1:(2-8):(3-15); more preferably, the mass ratio of the recombinant collagen, the polyhydroxyl compound, and the water is 1:(3-6):(5-12).

[0019] In some embodiments, the mass ratio of the emulsifier and the oil and fat is 1:(2-60); preferably, the mass ratio of the emulsifier and the oil and fat is 1:(3-55); more preferably, the mass ratio of the emulsifier and the oil and fat is 1:(5-50).

[0020] In some embodiments, the recombinant collagen has a mass percentage of 0.1-5%, the polyhydroxyl compound has a mass percentage of 0.3-15%, the oil has a mass percentage of 35-85%, the emulsifier has a mass percentage of 1-25%, and the water has a mass percentage of 2-20%.

[0021] In some embodiments, the recombinant collagen has a mass percentage of 0.3-5%, the polyhydroxyl compound has a mass percentage of 0.6-10%, the oil has a mass percentage of 40-85%, the emulsifier has a mass percentage of 1-20%, and the water has a mass percentage of 3-20%.

[0022] In some embodiments, the recombinant collagen has a mass percentage of 0.5-5%, the polyhydroxyl compound has a mass percentage of 1-15%, the oil has a mass percentage of 40-75%, the emulsifier has a mass percentage of 5-20%, and the water has a mass percentage of 5-20%.

[0023] In some embodiments, the recombinant collagen has a mass percentage of 1-5%, the polyhydroxyl compound has a mass percentage of 2-15%, the oil has a mass percentage of 65-75%, the emulsifier has a mass percentage of 10-20%, and the water has a mass percentage of 5-20%.

[0024] In some embodiments, the sum of the mass percentages of the recombinant collagen and the polyhydroxyl compound is < 28%; preferably, the sum of the mass percentages of the recombinant collagen and the polyhydroxyl compound is < 21%.

[0025] In some embodiments, the average particle size of the oil-soluble composition is 40-200 nm; preferably, the average particle size is 50-150 nm; more preferably, the average particle size is 60-120 nm.

[0026] In some embodiments, the recombinant collagen is one or more of type I, type III, type V, type VII, and type XVII.

[0027] In some embodiments, the recombinant collagen is type I.

[0028] In some other embodiments, the recombinant collagen is type III.

[0029] In some other embodiments, the recombinant collagen is type V.

[0030] In some other embodiments, the recombinant collagen is type VII.

[0031] In other embodiments, the recombinant collagen is type XVII.

[0032] In some embodiments, the polyhydroxy compound is selected from one or more of a boswellia, glucose, maltose, trehalose, mannose, sucrose, maltose, fructose. Preferably, the polyhydroxy compound is boswellia.

[0033] In some embodiments, the plant-derived oil is selected from one or more of white pool flower seed oil, olive oil, peanut oil, corn oil, coconut oil, camellia seed oil, soybean oil, wheat germ oil, oat kernel oil, avocado oil, macadamia seed oil, oil tea seed oil, jojoba seed oil, cocoa seed oil;

[0034] Preferably, the plant-derived oil is selected from one or more of soybean oil, camellia seed oil, white pool flower seed oil, jojoba seed oil.

[0035] In some embodiments, the synthetic oil is selected from one or more of caprylic / capric triglyceride, oleic acid triglyceride, lauric acid triglyceride, myristic acid triglyceride, dicaprylyl carbonate, neopentyl glycol dicaprylate / dicaprate, dioctyl ether, ethylhexyl palmitate, neopentyl glycol di(ethylhexanoate) ester, isopentyl laurate, octyldodecyl myristate, isopropyl isostearate, cocoyl caprylate / caprate, isononyl isononanoate;

[0036] Preferably, the synthetic oil is selected from one or more of caprylic / capric triglyceride, isononyl isononanoate.

[0037] In some embodiments, the hydrocarbon-based oil is selected from one or more of squalane, isooctane, isododecane, isohexadecane, isoeicosane, C 10 ~ C 13 isododecane, isohexadecane, isoeicosane; 12 ~ C 14 isododecane, isohexadecane, isoeicosane;

[0038] Preferably, the hydrocarbon-based oil is squalane.

[0039] In some embodiments, the polyethylene glycol (PEG) difatty acid ester is a polyethylene glycol (PEG) difatty acid ester having a degree of polymerization of 2 to 30; the fatty acid is selected from one or more of stearic acid, palmitic acid, oleic acid, isostearic acid, polyhydroxy stearic acid;

[0040] Preferably, the polyethylene glycol (PEG) difatty acid ester is selected from one or more of PEG-2 distearate, PEG-3 distearate, PEG-3 dipalmitate, PEG-4 distearate, PEG-4 dioleate, PEG-30-dipolyhydroxy stearate;

[0041] More preferably, the polyethylene glycol (PEG) difattyate is selected from one or more of PEG-3 distearate, PEG-3 dioleate, PEG-30-dipolyhydroxystearate.

[0042] In some embodiments, the polyglyceryl difattyate is a polyglyceryl difattyate having a degree of polymerization of 2 to 4; the fatty acid is selected from one or more of stearic acid, palmitic acid, oleic acid, isostearic acid, and polyhydroxy stearic acid;

[0043] Preferably, the polyglyceryl difattyate is a polyglyceryl difattyate having a degree of polymerization of 2 to 4; the fatty acid is selected from one or more of stearic acid, oleic acid, isostearic acid, isostearate / polyhydroxystearate / sebacate and polyhydroxy stearic acid;

[0044] More preferably, the polyglyceryl difattyate is selected from one or more of polyglyceryl-2 dioleate, polyglyceryl-2 distearate, polyglyceryl-2 distearate, polyglyceryl-2 dioleate, polyglyceryl-3 distearate, polyglyceryl-3 diisostearate, polyglyceryl-2 dipolyhydroxystearate, polyglyceryl-4 diisostearate / polyhydroxystearate / sebacate.

[0045] More preferably, the polyglyceryl difattyate is selected from one or more of polyglyceryl-2 dipolyhydroxystearate, polyglyceryl-2 dioleate, polyglyceryl-4 diisostearate / polyhydroxystearate / sebacate.

[0046] In some embodiments, the emulsifier is selected from one or more of PEG-2 distearate, PEG-3 distearate, PEG-3 dipalmitate, PEG-4 distearate, PEG-4 dioleate, PEG-30-dipolyhydroxystearate, polyglyceryl-2 dioleate, polyglyceryl-2 distearate, polyglyceryl-2 distearate, polyglyceryl-2 dioleate, polyglyceryl-3 distearate, polyglyceryl-3 diisostearate, polyglyceryl-2 dipolyhydroxystearate, polyglyceryl-4 diisostearate / polyhydroxystearate / sebacate.

[0047] Preferably, the emulsifier is selected from one or more of PEG-2 distearate, PEG-3 distearate, PEG-3 dipalmitate, PEG-30-dipolyhydroxystearate, polyglyceryl-2 dioleate, polyglyceryl-2 distearate, polyglyceryl-2 distearate, polyglyceryl-2 dioleate, polyglyceryl-2 dipolyhydroxystearate, polyglyceryl-4 diisostearate / polyhydroxystearate / sebacate.

[0048] In particular, in some embodiments, the emulsifier is PEG-2 distearate.

[0049] Specifically, in some embodiments, the emulsifier is polyglyceryl-2 distearate.

[0050] Specifically, in some embodiments, the emulsifier is polyglyceryl-2 dipolyhydroxystearate.

[0051] Specifically, in some embodiments, the emulsifier is polyglyceryl-4 diisostearate / polyhydroxystearate / sebacate.

[0052] Specifically, in some embodiments, the emulsifier is PEG-30-dipolyhydroxystearate.

[0053] The second aspect of the present application relates to a method for preparing the aforementioned oil-soluble composition, the method comprising the steps of:

[0054] (1) adding an emulsifier to oil and dissolving it into an oil phase; adding recombinant collagen and polyols into water and dissolving them into an aqueous phase;

[0055] (2) Slowly add the aqueous phase to the oil phase during stirring and continue stirring to obtain an oil-soluble composition.

[0056] Wherein, in the oil-soluble composition, the mass percentage of recombinant collagen is 0.1%-5%; the mass percentage of polyol is 1%-15%; the mass percentage of oil is 30-90%; the mass percentage of emulsifier is 1-30%; and the mass percentage of water is 1%-20%.

[0057] The present invention brings about one or more of the following technical effects:

[0058] 1. Recombinant collagen has good water solubility, making it difficult to incorporate into oil phases. This limits its use in oil-phase products such as essences, massage oils, hair oils, and cosmetics. The oil-soluble composition provided by the present invention, by encapsulating recombinant collagen within a suitable oil phase, can increase the amount of recombinant collagen added to 5%. This composition is miscible with oils and fats in any proportion, meeting the development needs for the addition of recombinant collagen to oil-phase cosmetics.

[0059] 2. Transparent products are more in line with consumer preferences, thus placing higher technical demands on the transparency and stability of oil-soluble compositions. With increasing amounts of recombinant collagen, for example, up to 1% (wt%), the prepared recombinant collagen oil-soluble composition can become turbid and demulsify after prolonged storage and exposure to high temperatures. Through screening experiments, the present invention unexpectedly discovered that adding polyols to the aqueous phase can effectively address this turbidity and demulsification problem in recombinant collagen oil-soluble compositions.

[0060] 3. In order to solve the stability problem of the high additive amount of the oil-soluble recombinant collagen composition, it is necessary to add a higher mass ratio of polyhydroxy compound in the aqueous phase, and when the additive amount of the recombinant collagen is 5%, 15% of the polyhydroxy compound needs to be added, and it is difficult to achieve the transparent and stable state of the oil-soluble composition with high drug loading amount, and the application accidentally found that some emulsifiers can provide the oil-soluble composition which can obtain a stable and transparent oil-soluble composition even in the case of high drug loading amount.

[0061] 4. The oil-soluble composition provided by the application has a transparent appearance, a particle size of about 100 nm, and can be stably placed for a long time under the condition of 2-8 DEG C to 48 DEG C, and has good adaptability and dilution stability. After dilution with different oils in different proportions, it is still stable when placed in an environment of 2-8 DEG C to 48 DEG C.

[0062] 5. The water-in-oil emulsion composition provided by the application has a simple preparation process and is prepared by simple stirring at room temperature, and is easy to industrialize.

[0063] The details of the application are set forth in the accompanying description below. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, illustrative methods and materials are described herein, and it is to be understood that similar or equivalent methods and materials are intended to be encompassed by the present application, according to the description and claims herein. In the description and the appended claims, the singular forms also include the plural unless the context clearly dictates otherwise. Unless defined otherwise, 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. All patents and publications cited in this specification are incorporated herein by reference in their entirety.

[0064] The contents of all references (including literature references, issued patents, published patent applications, and co-pending patent applications) cited throughout this application are hereby expressly incorporated by reference. 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. BRIEF DESCRIPTION OF DRAWINGS

[0065] Figure 1 Particle size spectrum of the oil-soluble composition of an embodiment of the application. DETAILED DESCRIPTION

[0066] The technical solutions of the application are further described below in combination with some specific embodiments, but these embodiments do not limit the scope of the application.

[0067] Example 1 Preparation of recombinant collagen oil-soluble composition

[0068] (1) Preparation of oil phase

[0069] Take a 100ml beaker, add 1g PEG-2 distearate and 40g caprylic / capric triglyceride respectively, and stir at room temperature until dissolved evenly to obtain the oil phase;

[0070] (2) Preparation of aqueous phase

[0071] Take a 10ml centrifuge tube, add 1g recombinant type I collagen, 3g bosin, and 5g pure water respectively, heat and dissolve to obtain the aqueous phase;

[0072] (3) Preparation of oil-soluble composition of recombinant collagen

[0073] Under magnetic stirring at 100 rpm / min, the aqueous phase was added dropwise to the oil phase and stirred for 10 minutes until the solution became clear and transparent, thereby obtaining a 2% recombinant type I collagen oil-soluble composition.

[0074] Example 2 Particle size determination of oil-soluble composition

[0075] This embodiment provides a method for measuring the particle size of a recombinant collagen oil-soluble composition. The particle size measurement method is exemplified as follows:

[0076] Take 20 μL of the 2% recombinant type I collagen oil-in-water emulsion prepared in Example 1 and dissolve it in 1 mL of caprylic / capric triglyceride. Use Malvern nanoparticle size analyzer (Zetasizer Pro) to measure the sample. The particle size spectrum is shown in Figure 1 . Figure 1 The average particle size of the 2% recombinant collagen oil-soluble composition was 89 nm.

[0077] Example 3 Effects of different types of polyols on the stability and transparency of oil-soluble compositions of recombinant collagen

[0078] The purpose of this example is to investigate the effects of adding different types of polyhydroxy compounds on the stability and transparency of a high-content recombinant collagen oil-soluble composition. The preparation methods of each formulation in this example refer to Example 1.

[0079] Inspection method: Observe the appearance of each formula and record the results. The results are shown in Table 1.

[0080] Table 1 Effects of different types of polyols on the stability of oil-soluble compositions of recombinant collagen

[0081]

[0082]

[0083] The oil liquid prepared from Formulation 1.1 and Formulation 1.2 containing less than 0.5% of recombinant collagen and without adding polyhydroxy compounds is translucent, and has no obvious change in appearance when left at room temperature (25°C, 7 days) or at high temperature (48°C, 1 day), and has good stability and transparency. However, when the amount of recombinant collagen is more than 0.5%, the oil liquid prepared is turbid, has poor transparency, and appears to be demulsified and layered in the high temperature test (48°C, 1 day), and has poor stability, such as the oil liquid prepared from Formulation 1.3 and Formulation 1.4. In order to improve the transparency of the oil-soluble composition, it is unexpectedly found in this test that the transparency of the oil-soluble composition is obviously improved after adding several polyhydroxy compounds, such as bovine growth factor, glucose, trehalose, mannose, sucrose, maltose and fructose, on the basis of Formulation 1.4, and the effect of bovine growth factor is particularly significant, and the oil-soluble composition has better stability in the high temperature test (48°C, 1 day). However, the effect of polyhydroxy compounds, such as propylene glycol, glycerol and mannitol, is not good in Formulation 1.5-Formulation 1.10, and the appearance is turbid.

[0084] Example 4: Effect of different contents of polyhydroxy compounds on the stability of the oil-soluble composition of recombinant collagen

[0085] This example mainly investigates the effect of different contents of polyhydroxy compounds on the stability of the oil-soluble composition of recombinant collagen, and the preparation method of each formulation in this example is referred to Example 1.

[0086] Investigation method: the appearance of each formulation is observed and recorded. The results are shown in Table 2.

[0087] Table 2: Effect of different contents of polyhydroxy compounds on the stability of the oil-soluble composition of recombinant collagen

[0088]

[0089] The results in Table 2 show that when the mass ratio of polyhydroxy compound to recombinant collagen is ≥3:1, the oil-soluble composition of recombinant collagen has good stability.

[0090] Example 5: Effect of different types of emulsifiers on the stability of the oil-soluble composition of recombinant collagen

[0091] In preparing the oil-soluble composition of recombinant collagen, emulsifiers are the key factor in forming the oil-soluble composition. Generally, the hydrophilic-lipophilic balance (HLB) of the emulsifiers that can form the oil-soluble composition is generally 3-8, and these emulsifiers can be classified into different types of emulsifiers based on the nature of the hydrophilic group, such as non-ionic emulsifiers, cationic emulsifiers, and anionic emulsifiers. The emulsifiers used in general cosmetics are mainly non-ionic emulsifiers, and the main reason is that non-ionic emulsifiers can tolerate a wider pH range and are not affected by electrolytes.

[0092] The purpose of this example is to investigate the effect of different types of non-ionic emulsifiers on the stability of the oil-soluble composition of recombinant collagen.

[0093] The preparation method of each formula in this example is referred to Example 1.

[0094] Investigation method: observe the appearance of each formula and record. The results are shown in Table 3.

[0095] Table 3 Effect of different types of non-ionic emulsifiers on the stability of the oil-soluble composition of recombinant collagen

[0096]

[0097] As shown in Table 3, different types of emulsifiers have a significant difference in the stability of the oil-soluble composition of recombinant collagen. Formulas 3.1-3.2 and 3.5-3.6 all show demulsification and layering, and emulsions cannot be prepared; while formulas 3.3-3.4 and formulas 3.7-3.10 all produce semi-transparent stable oil liquid. It is speculated that the reason may be that the high content of water-soluble active substances in the water phase can disturb the oil-water interface, resulting in unstable oil-soluble composition, while polyethylene glycol (PEG) di-fatty acid ester and polyglycerol di-fatty acid ester can more firmly stabilize the oil-water interface.

[0098] Example 6 Effect of different emulsifiers on the oil-soluble composition of recombinant collagen

[0099] This example further investigates the effect of different types of double-chain emulsifiers on the oil-soluble composition. In this example, the preparation method of each formula is referred to Example 1, and the particle size determination method is referred to Example 2.

[0100] Investigation method: observe the appearance of each formula and record. The results are shown in Table 4.

[0101] Table 4 Effect of different emulsifiers on the oil-soluble composition of recombinant collagen

[0102]

[0103]

[0104] The results in Table 4 show that the longer the lipophilic chain of the emulsifier, the smaller the particle size of the emulsion formed, and the higher the transparency of the emulsion. It is speculated that the reason is that the longer the lipophilic chain, the more stable the oil-water interface formed, thereby forming smaller particle size emulsion.

[0105] Example 7 Formulation screening of different concentrations of oil-soluble recombinant collagen composition

[0106] In order to improve the drug loading capacity of the oil-soluble composition while ensuring the transparency of the oil-soluble composition, the amount of PEG-30-dihydroxyl stearate and polyglyceryl-2 dihydroxystearate, two emulsifiers, and other factors were comprehensively screened in this embodiment. The specific results are shown in Tables 5 and 6. The preparation method of each formulation is referred to Example 1.

[0107] Investigation method: observe the appearance of each formulation and record it.

[0108] Table 5 Effect of polyglyceryl-2 dihydroxystearate on different concentrations of oil-soluble recombinant collagen composition

[0109]

[0110] Table 6 Effect of PEG-30-dihydroxystearate on different concentrations of oil-soluble recombinant collagen composition

[0111]

[0112] From Tables 5 and 6, it can be seen that when polyglyceryl-2 dihydroxystearate and PEG-30-dihydroxystearate are used as emulsifiers, when the sum of the mass percentages of recombinant collagen and bovine growth hormone reaches a certain limit (for example, 28%), the appearance of the oil composition cannot reach the transparent effect, and appears turbid and milky white appearance; when the sum of the mass percentages of recombinant collagen and bovine growth hormone is ≤21% (especially ≤20%), a good transparent oil liquid can be obtained. Among the two emulsifiers, PEG-30-dihydroxystearate has the best emulsifying effect. When the content of recombinant collagen is 1%-5% and the content of bovine growth hormone is 3%-15%, 5%-20% PEG-30-dihydroxystearate can achieve complete transparency or basic transparency.

[0113] Example 8 Adaptability of PEG-30-dihydroxystearate in different oils

[0114] In this embodiment, the effect of using various different oils on the appearance of the oil-soluble composition was investigated when PEG-30-dihydroxystearate was used as an emulsifier. The preparation method of each formulation is referred to Example 1. The specific results are shown in Table 7.

[0115] Investigation method: observe the appearance of each formula and record.

[0116] Table 7 Transparency of PEG-30-dihydroxyl hardened ester in different oils

[0117]

[0118] As shown in Table 7, the oil-soluble compositions prepared by various formulas all have the effect of being substantially transparent or completely transparent, which shows that PEG-30-dihydroxyl hardened ester has good adaptability to typical various oils.

[0119] Example 9 Investigation of adaptability of different types of recombinant collagen

[0120] This example investigates the adaptability of PEG-30-dihydroxyl hardened ester as an emulsifier and bovine collagen as a polyhydroxyl compound to different types of recombinant collagen.

[0121] The preparation method of the formula is referred to Example 1. The specific results are shown in Table 8.

[0122] Investigation method: observe the appearance of each formula and record.

[0123] Table 8 Oil-soluble compositions of different types of recombinant collagen

[0124]

[0125] As shown in Table 8, the oil-soluble compositions prepared by various formulas all have the effect of being substantially transparent or completely transparent, which shows that the composition system has good adaptability to different types of recombinant collagen.

[0126] Example 10 Investigation of stability of high drug loading recombinant collagen oil-soluble composition

[0127] This example investigates the stability of water-in-oil emulsions of different formula proportions of recombinant collagen with PEG-30-dihydroxyl hardened ester as an emulsifier at 2-8℃, 25±2℃ and 48±2℃. The preparation method of each emulsion is referred to Example 1, and the particle size determination method is referred to Example 2.

[0128] Table 9 Investigation of stability of recombinant collagen oil-soluble composition at 2-8℃

[0129]

[0130] Table 10 Investigation of stability of recombinant collagen oil-soluble composition at 25±2℃

[0131]

[0132] Table 11 Stability of the recombinant collagen oil-soluble composition at 48±2°C

[0133]

[0134] As shown in Tables 9, 10 and 11, the particle size and appearance of the recombinant collagen oil-soluble composition with different proportions did not change significantly after being placed at 2-8°C, 25±2°C and 48±2°C for 1 week, and remained transparent or translucent, indicating good stability.

[0135] Example 11 Dilution and stability of the 2% recombinant collagen oil-soluble composition

[0136] The 2% recombinant collagen oil-soluble composition was diluted and mixed with camellia seed oil, soybean oil, caprylic / capric acid triglyceride, dicaprylyl carbonate, isononyl isononanoate, neopentyl glycol dioctanoate / dicaprate, dicaprylyl ether, ethylhexyl palmitate, squalane and isohexadecane in different proportions, and the stability was investigated at 2-48°C. The observation index was whether the appearance showed demulsification or turbidity.

[0137] Table 12 Stability of the 2% recombinant collagen oil-soluble composition mixed with oil 1:9

[0138]

[0139] Table 13 Stability of the 2% recombinant collagen oil-soluble composition mixed with oil 3:7

[0140]

[0141] Table 14 Stability of the 2% recombinant collagen oil-soluble composition mixed with oil 5:5

[0142]

[0143] Table 15 Stability of the 2% recombinant collagen oil-soluble composition mixed with oil 9:1

[0144]

[0145] As shown in Tables 12, 13, 14 and 15, the 2% recombinant collagen oil-soluble composition mixed with different types of oil in different proportions did not change significantly in appearance after being placed at 2-8°C, 25±2°C and 48±2°C for 30 days, and remained transparent or translucent, indicating good adaptability and stability.

[0146] It should be understood that after reading the above content of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. An oil-soluble composition, characterized in that The composition comprises recombinant collagen, polyol, oil, emulsifier and water; wherein the mass percentage of recombinant collagen is 0.1%-5%; the mass percentage of polyol is 0.3%-15%; the mass percentage of oil is 30-90%; the mass percentage of emulsifier is 1-30%; and the mass percentage of water is 1%-20%. The polyhydroxy compound is selected from one or more of bosacene, glucose, maltose, trehalose, mannose, sucrose, maltose, and fructose; The oil is selected from one or more of plant-derived oils, synthetic oils and hydrocarbon oils; The emulsifier is selected from polyethylene glycol difatty acid esters and / or polyglycerol difatty acid esters; the fatty acid is selected from one or more of stearic acid, palmitic acid, oleic acid, isostearic acid, and polyhydroxystearic acid.

2. The composition according to claim 1, characterized in that The mass percentage of the recombinant collagen is 0.3%-5%; preferably, the mass percentage of the recombinant collagen is 0.5%-5%; more preferably, the mass percentage of the recombinant collagen is 1%-5%.

3. The composition according to claim 1, characterized in that The mass percentage of the recombinant collagen is 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 1%, 1.5%, 2%, 2.5%, 3% or 5%.

4. The composition according to claim 1, characterized in that The mass percentage of the polyhydroxy compound is 0.6%-15%; preferably, the mass percentage of the recombinant collagen is 1.5%-10%; more preferably, the mass percentage of the recombinant collagen is 3%-10%.

5. The composition according to claim 1, characterized in that The mass percentage of the recombinant collagen is 0.6%, 1%, 1.5%, 2%, 3%, 5%, 8% or 10% of the mass percentage of the polyhydroxy compound.

6. The composition according to claim 1, characterized in that The mass percentage of the oil is 40-85%; preferably, the mass percentage of the oil is 40-75%.

7. The composition according to claim 1, characterized in that The mass percentage of the oil is 40%, 45%, 50%, 55%, 60%, 65%, 70% or 75%.

8. The composition according to claim 1, characterized in that The mass percentage of the emulsifier is 1-25%; preferably, the mass percentage of the emulsifier is 1-20%; more preferably, the mass percentage of the emulsifier is 5-20%.

9. The composition according to claim 1, characterized in that The mass percentage of the emulsifier is 5%, 10%, 15% or 20%.

10. The composition according to claim 1, characterized in that The mass percentage of the water is 3%-20%; preferably, the mass percentage of the water is 5%-20%.

11. The composition according to claim 1, characterized in that The mass percentage of the water is 5%, 10%, 15% or 20%.

12. The composition according to claim 1, characterized in that The mass ratio of the recombinant collagen, polyol and water is 1:(1.5-10):(1-20); preferably, the mass ratio of the recombinant collagen, polyol and water is 1:(2-8):(3-15); more preferably, the mass ratio of the recombinant collagen, polyol and water is 1:(3-6):(5-12).

13. The composition according to claim 1, characterized in that The mass ratio of the emulsifier to the oil is 1:(2-60); preferably, the mass ratio of the emulsifier to the oil is 1:(3-55); more preferably, the mass ratio of the emulsifier to the oil is 1:(5-50).

14. The composition according to claim 1, characterized in that The mass percentage of the recombinant collagen is 0.1%-5%, the mass percentage of the polyol is 0.3-15%, the mass percentage of the oil is 35-85%, the mass percentage of the emulsifier is 1-25%, and the mass percentage of the water is 2%-20%; preferably, the mass percentage of the recombinant collagen is 0.3%-5%, the mass percentage of the polyol is 0.6-10%, the mass percentage of the oil is 40-85%, the mass percentage of the emulsifier is 1-20%, and the mass percentage of the water is 3%-20%; More preferably, the mass percentage of the recombinant collagen is 0.5%-5%, the mass percentage of the polyol is 1-15%, the mass percentage of the oil is 40-75%, the mass percentage of the emulsifier is 5-20%, and the mass percentage of the water is 5%-20%; more preferably, the mass percentage of the recombinant collagen is 1%-5%, the mass percentage of the polyol is 2-15%, the mass percentage of the oil is 65-75%, the mass percentage of the emulsifier is 10-20%, and the mass percentage of the water is 5%-20%.

15. The composition according to claim 1, characterized in that The sum of the mass percentages of the recombinant collagen and the polyhydroxy compound is less than 28%; preferably, the sum of the mass percentages of the recombinant collagen and the polyhydroxy compound is ≤ 21%.

16. The composition according to claim 1, characterized in that The average particle size of the oil-soluble composition is 40-200 nm; preferably, the average particle size is 50-150 nm; more preferably, the average particle size is 60-120 nm.

17. The composition according to claim 1, characterized in that The recombinant collagen is one or more of type I, type III, type V, type VII, and type XVII.

18. The composition according to any one of claims 1 to 17, characterized in that The plant-derived oil is selected from one or more of meadowfoam seed oil, olive oil, peanut oil, corn oil, coconut oil, camellia seed oil, soybean oil, wheat germ oil, oat kernel oil, avocado oil, macadamia seed oil, camellia seed oil, jojoba seed oil, and cocoa seed oil; preferably, the plant-derived oil is selected from one or more of soybean oil, camellia seed oil, meadowfoam seed oil, and jojoba seed oil.

19. The composition according to any one of claims 1 to 17, characterized in that The synthetic oil is selected from one or more of caprylic / capric triglyceride, oleic triglyceride, lauric triglyceride, myristic triglyceride, dicaprylyl carbonate, neopentyl glycol dicapryl / dicaprate, dicaprylyl ether, ethylhexyl palmitate, neopentyl glycol di(ethylhexanoate), isoamyl laurate, octyldodecanol myristyl ester, isopropyl isostearate, coconut oil-caprylate / caprate, and isononyl isononanoate; preferably, the synthetic oil is selected from one or more of caprylic / capric triglyceride and isononyl isononanoate.

20. The composition according to any one of claims 1 to 17, characterized in that The hydrocarbon oil is selected from squalane, isooctane, isododecane, isohexadecane, isoeicosane, C 10 ~C 13 Isoparaffins, C 12 ~C 14 Isoparaffin; preferably, the hydrocarbon oil is squalane.

21. The composition according to any one of claims 1 to 17, characterized in that The polyethylene glycol difatty acid ester is a polyethylene glycol difatty acid ester with a degree of polymerization of 2 to 30; the fatty acid is selected from one or more of stearic acid, palmitic acid, oleic acid, isostearic acid, and polyhydroxystearic acid; Preferably, the polyethylene glycol difatty acid ester is selected from one or more of PEG-2 distearate, PEG-3 distearate, PEG-3 dipalmitate, PEG-4 distearate, PEG-4 dioleate, and PEG-30-dipolyhydroxystearate; Preferably, the polyethylene glycol difatty acid ester is selected from one or more of PEG-3 distearate, PEG-3 dioleate, and PEG-30-dipolyhydroxystearate.

22. The composition according to any one of claims 1 to 17, characterized in that The polyglycerol difatty acid ester is a polyglycerol difatty acid ester with a degree of polymerization of 2 to 4; the fatty acid is selected from one or more of stearic acid, palmitic acid, oleic acid, isostearic acid, and polyhydroxystearic acid; Preferably, the polyglycerol difatty acid ester is a polyglycerol difatty acid ester with a degree of polymerization of 2 to 4; the fatty acid is selected from one or more of stearic acid, oleic acid, isostearic acid, isostearate / polyhydroxystearate / sebacate and polyhydroxystearic acid; Preferably, the polyglyceryl difatty acid ester is selected from one or more of polyglyceryl-2 dioleate, polyglyceryl-2 distearate, polyglyceryl-2 distearate, polyglyceryl-2 dioleate, polyglyceryl-3 distearate, polyglyceryl-3 diisostearate, polyglyceryl-2 dipolyhydroxystearate, and polyglyceryl-4 diisostearate / polyhydroxystearate / sebacate; More preferably, the polyglyceryl difatty acid ester is selected from one or more of polyglyceryl-2 dipolyhydroxystearate, polyglyceryl-2 dioleate, polyglyceryl-4 diisostearate / polyhydroxystearate / sebacate.

23. The composition according to any one of claims 1 to 17, characterized in that The emulsifier is selected from one or more of PEG-2 distearate, PEG-3 distearate, PEG-3 dipalmitate, PEG-4 distearate, PEG-4 dioleate, PEG-30-dipolyhydroxystearate, polyglyceryl-2 dioleate, polyglyceryl-2 distearate, polyglyceryl-3 distearate, polyglyceryl-3 diisostearate, polyglyceryl-2 dipolyhydroxystearate, polyglyceryl-4 diisostearate / polyhydroxystearate / sebacate; Preferably, the emulsifier is selected from one or more of PEG-2 distearate, PEG-3 distearate, PEG-3 dipalmitate, PEG-30-dipolyhydroxystearate, polyglyceryl-2 dioleate, polyglyceryl-2 distearate, polyglyceryl-2 dipolyhydroxystearate, polyglyceryl-4 diisostearate / polyhydroxystearate / sebacate; Preferably, the emulsifier is PEG-2 distearate; Preferably, the emulsifier is polyglyceryl-2 distearate; Preferably, the emulsifier is polyglyceryl-2 dipolyhydroxystearate; Preferably, the emulsifier is polyglyceryl-4 diisostearate / polyhydroxystearate / sebacate; Preferably, in some embodiments, the emulsifier is PEG-30-dipolyhydroxystearate.

24. A method for preparing the oil-soluble composition according to any one of claims 1 to 23, comprising the steps of: (1) adding an emulsifier to oil and dissolving it into an oil phase; adding recombinant collagen and polyols into water and dissolving them into an aqueous phase; (2) Slowly add the aqueous phase to the oil phase during stirring and continue stirring to obtain an oil-soluble composition.

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