Alkaloid liposomes, process for their preparation and use in external preparations

Abstract

The application discloses an alkaloid liposome and a preparation process and application thereof in external preparations, and belongs to the technical field of external preparations. The technical problem to be solved is to provide an alkaloid liposome technology with higher stability and safety and no irritation. The scheme points are that raw materials include alkaloids, hydrogenated lecithin, oil, polyhydric alcohol and water; the alkaloids are selected from at least three kinds of piperine, tetrahydropiperine, fumarine, dihydroavenanthramide D, caffeine and leonuride; the hydrogenated lecithin is selected from at least one of PHOSPHOLIPON 80H or Phoslip HPC. The liposome has good stability and low irritation, and has the effects of promoting local tissue microcirculation, reducing resistance, improving local inflammation, improving local body temperature, enhancing tissue metabolism and the like when being used externally on the skin.

Description

Technical Field

[0001] This invention relates to the field of topical formulation technology, and more specifically, to alkaloid liposomes, their preparation process, and their application in topical formulations. Background Technology

[0002] In daily life, prolonged exposure to low temperatures from air conditioning, sedentary work habits, and seasonal damp and cold weather can easily lead to sub-health conditions such as cold extremities, aversion to cold, and fatigue. In traditional Chinese medicine, these symptoms are often categorized as "insufficient Yang Qi" or "stagnation of cold and dampness." To alleviate these discomforts, many folk remedies rooted in natural products have been developed. For example, in Hunan and Sichuan, where damp and cold climates are prevalent, people often use pungent ingredients like Sichuan peppercorns and chili peppers to warm and dispel cold. In southern China, there is a tradition of drinking pepper and pork tripe soup and using pepper roots to make soup and stews, all to dispel cold and dampness, warm the body, and relieve pain. Tea and coffee, widely consumed in daily life, are also considered daily beverages that refresh the mind and promote blood circulation. Modern research reveals that the ability of these dietary and conditioning practices to "warm and dispel cold, promote Qi and blood circulation" is closely related to the various alkaloid active ingredients contained in their plant materials—such as piperine in pepper and caffeine in coffee and tea. These components have clear physiological activities in promoting microcirculation, increasing local temperature and tissue metabolism, thus providing an intrinsic scientific basis for the traditional practice of "prevention of disease" and physical conditioning.

[0003] Alkaloids are a class of nitrogen-containing basic organic lead compounds widely found in plants (and a few in animals). Because their properties are similar to those of alkaloids, they are sometimes called "pseudo-alkaloids." They are diverse and structurally complex, mainly distributed in legumes, solanaceae, piperines, and grasses. As important secondary metabolites and natural active ingredients of medicinal plants, alkaloids exhibit significant and diverse pharmacological activities, including anti-inflammatory, antioxidant, anti-fatigue, analgesic, immunomodulatory, and antitumor effects, and have important value in the clinical treatment of various diseases. Studies have shown that alkaloids can effectively promote tissue microcirculation, reduce blood flow resistance, and improve local obstruction; they can also increase local temperature and enhance tissue metabolic activity, thereby accelerating inflammation resolution and tissue repair. Therefore, alkaloids, as important natural plant-derived lead compounds, have potential applications in various body care products and cosmetics.

[0004] However, most alkaloids have poor water solubility and are unstable, resulting in low bioavailability and difficulties in formulation, thus limiting their application in body care products and cosmetics. For example, piperine has poor water solubility and is easily converted and degraded under acidic, alkaline, light, heat, and oxygen conditions; corydaline is poorly soluble in water and easily decomposes and inactivates under photothermal effects; caffeine is easily inactivated under high temperature, weak alkaline, and enzymatic environments; leonurine also suffers from low solubility and sensitivity to photothermal effects; dihydroamarine D and tetrahydropiperine are poorly soluble in water. On the other hand, to achieve true efficacy in various cosmetics and body care products, in addition to high molecular activity concentration, it is also necessary to quickly overcome the osmotic barrier to deliver the active substances to the target site.

[0005] Lipid carriers are substances containing active ingredients encapsulated in lipid materials to prevent the active substances from being destroyed by the environment (temperature, oxygen, pH, enzymes, etc.), thereby improving the bioactivity and utilization rate of the active substances. In addition, lipid carriers have advantages such as high bioavailability, good absorption, and high stability, and are widely used in medicine, cosmetics, food, and genetic engineering.

[0006] Relevant patent documents retrieved:

[0007] This document, published in China (CN1446534A) on October 8, 2003, discloses a liposome containing alkaloids from traditional Chinese medicine and its preparations. It can increase the effective blood drug concentration. The components and contents are: 0.1-2% alkaloids from traditional Chinese medicine, 0.5-10% phospholipids, and 0-5% cholesterol (all by mass percentage). The phospholipids are lecithin, distearate phosphatidylcholine, dipalmitoylphosphatidylcholine, dioleoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, and distearate phosphatidylglycerol. The alkaloids include berberine, berberine, berberine alkaloids, tetrahydropalmatine, and berberine.

[0008] The document, published in China (CN119112694A) on December 13, 2024, discloses a tri-factor liposome, its preparation process, and its application in a skin-nourishing cushion cream. The tri-factor liposome comprises the following components in the indicated mass fractions: hydrogenated lecithin 0.5-4%, glycerin 30-70%, oil 5-15%, cholesterol 0.1-0.5%, prosciuttoin 1-10%, ectoine 1-10%, ergothioneine 0.05-0.5%, bisabolol 0.1-1%, auxiliary agents 0.1-1%, and solvent balance.

[0009] Relevant non-patent literature retrieved:

[0010] The journal or book title is "Pharmaceutical and Clinical Research," the document title is "Preparation Technology of Alkaloid Liposomes," volume number 3, issue number 17, and the publication date is January 13, 2009. This document discloses the research progress of conventional preparation processes of alkaloid liposomes (such as thin film method, ultrasonic dispersion method, reverse phase evaporation method, and active drug loading method) as well as novel liposome preparation technologies (combined application of preparation technologies to novel liposomes, preparation of surface-modified liposomes, and preparation technology of composite phospholipid liposomes).

[0011] The prior art represented by the aforementioned documents has at least the following unresolved technical problems or defects:

[0012] There are no solutions to the problems of poor solubility and instability of certain alkaloids, and their applicability in topical formulations has not been verified. Therefore, developing a novel nanodelivery system with both high water solubility and high bioavailability is of great significance to overcome the application of poorly soluble alkaloid active ingredients in topical formulations.

[0013] In solving the above problems or overcoming the above defects, the present invention encountered the following difficulties and obstacles:

[0014] 1. Since liposomes are to be used in topical formulations, the developed liposomes should meet the corresponding stability requirements. However, the stability requirements of topical formulations are quite stringent for liposomes. Therefore, detailed and long-term process investigation and exploration are usually required. In this process, we often encounter problems such as poor liposome particle size stability, large particle size changes under extreme conditions, such as easy stratification at 45℃, partial freezing of samples at -15℃, destruction of liposome structure after freeze-thaw, resulting in instability, and instability under centrifugation conditions.

[0015] 2. There are many types of topical body care formulations, the most common of which are creams and serums. In aqueous systems such as serums, liposomes are essentially diluted, and these systems usually have low viscosity, which can easily cause instability in the structure of liposomes. Therefore, when developing liposomes, it is necessary to take into account their stable application in various topical body care formulations.

[0016] 3. Microbial contamination of samples is difficult to handle. The addition of some preservatives can lead to instability in the liposome structure. Alternatively, the compatibility of the preservatives themselves or the difficulty in using them (such as the discoloration caused by using octanoyl hydroxamic acid with amino acid raw materials, and precipitation caused by increased p-hydroxyacetophenone content) can limit the application of liposome formulations. Therefore, a comprehensive consideration is needed when selecting preservatives.

[0017] 4. Current topical formulations mostly focus on basic skincare functions such as moisturizing and hydrating, and still have significant limitations in achieving biological effects such as promoting microcirculation, improving local metabolism, increasing local temperature, enhancing metabolism, and alleviating inflammation. As the largest organ in the human body, the skin has the potential to regulate physiological functions through local drug delivery, but existing formulations still face challenges in the efficient penetration and delivery of active ingredients, especially for alkaloids. While these alkaloids exhibit significant pharmacological activity, they also suffer from low solubility, poor stability, and insufficient bioavailability. Specifically: ① The skin's natural barrier severely limits the transdermal efficiency of active ingredients; ② Alkaloids have low solubility and poor chemical stability, easily degrading or precipitating, requiring advanced formulation technology for effective application; however, their inherent properties significantly affect the stability and final efficacy of the formulation; ③ These ingredients also pose certain risks of irritation and sensitization. These problems place higher demands on formulation development and technological optimization. Summary of the Invention

[0018] The purpose of this invention is to provide:

[0019] This invention provides an alkaloid liposome and related technologies, including a more stable, safe, and non-irritating alkaloid liposome and other technologies or combinations thereof.

[0020] Terminology Explanation:

[0021] Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0022] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0023] The definition of standard chemical terms can be found in the reference "Pharmacopoeia of the People's Republic of China (2020 Edition): China Medical Science and Technology Press: May 2020: First Edition".

[0024] Unless otherwise stated, conventional methods within the scope of the art, such as mixing methods, shall be used.

[0025] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0026] As used in this article, "mixing" refers to the process of uniformly distributing two or more components through manipulation, with the aim of achieving consistency in component content. For example, in pharmaceutics, mixing involves combined operations of solids with solids, solids with liquids, or liquids with liquids to ensure the homogeneity of the final product.

[0027] The term "shear rate" used in this article refers to a core concept in rheology, used to quantify the deformation rate of a fluid under shear force. Specifically, it is defined as the ratio of the flow velocity in the shear direction to the thickness of the laminar flow field in a laminar flow field. The unit is usually one second (s) -1 Strain rate (or strain rate) is calculated based on the velocity gradient, where du represents the velocity difference between adjacent fluid layers and dy represents the vertical distance between the two layers. This definition is applicable to common scenarios such as parallel plate flow and can be directly measured using experimental equipment such as shear meters or rheometers.

[0028] In a first aspect, the present invention provides: an alkaloid liposome, comprising: alkaloids, hydrogenated lecithin, oils, polyols, and water; wherein the alkaloids are selected from at least three of piperine, tetrahydropiperine, corydaline, dihydrooat alkaloid D, caffeine, and leonine; and wherein the hydrogenated lecithin is selected from PHOSPHOLIPON. ® 80H or Phoslip TM At least one of HPCs.

[0029] The technical effects of this invention can be achieved by any combination of three or more of the following alkaloids: piperine, tetrahydropiperine, corydaline, dihydroamarine D, caffeine, and leonurine.

[0030] The alkaloids are preferably a mixture of tetrahydropiperine, caffeine and leonurine.

[0031] Preferably, the mass ratio of the tetrahydropiperine, caffeine and leonurine is 1-3:1-3:4-10.

[0032] Any point value or any range of two point values ​​within the above range can achieve the technical effect of the present invention: including but not limited to 1:1:4, 1:2:5, 1:3:6, 1:1:7, 1:2:8, 2:3:9, and 3:1:10.

[0033] More preferably, the mass ratio of the tetrahydropiperine, caffeine and leonurine is 1-2:1-3:4-7.

[0034] More preferably, the mass ratio of the tetrahydropiperine, caffeine and leonurine is 2:1:7.

[0035] The oils are selected from at least one of the following: caprylic / capric triglyceride, squalane, egg yolk oil, jojoba seed oil, isononyl isononanoate, and phytosterol / octyldiol lauroyl glutamate.

[0036] The preferred oil is at least one of caprylic / capric triglyceride and squalane.

[0037] The polyol is selected from at least one of glycerol, propylene glycol, and butylene glycol.

[0038] The polyol is preferably at least one of glycerol and propylene glycol.

[0039] The raw materials are selected from the following parts by mass: the alkaloid liposomes include the following raw materials by mass: 1-12 parts alkaloids, 1-8 parts hydrogenated lecithin, 8-28 parts oils, 35-75 parts polyols and 10-30 parts water.

[0040] Any point value or any range of two point values ​​within the above range can achieve the technical effect of this invention:

[0041] For example, 1-12 parts of alkaloids represent the mass parts of alkaloids, including but not limited to: 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, and 12 parts, which will not be elaborated here.

[0042] For example, 1-8 parts of hydrogenated lecithin represent the following mass fractions of hydrogenated lecithin, including but not limited to: 1 part, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts, 5 parts, 5.5 parts, 6.0 parts, 6.5 parts, 7.0 parts, 7.5 parts, and 8.0 parts. These will not be elaborated further here.

[0043] For example, 8-28 parts of oil represents the number of parts of oil by weight, including but not limited to: 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, and 28 parts, which will not be elaborated further here.

[0044] For example, the mass fractions of 35-75 parts of polyol include, but are not limited to: 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, and 75 parts, which will not be elaborated further here.

[0045] For example, 10-30 parts water represents the following mass fractions of water, including but not limited to: 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30 parts, which will not be elaborated further here.

[0046] The preferred mass fractions of the raw materials are as follows: the alkaloid liposomes, by mass fractions, include the following raw materials: 1-10 parts alkaloids, 1-5 parts hydrogenated lecithin, 10-25 parts oils, 40-70 parts polyols and 18-20 parts water.

[0047] Any point value within the above range can achieve the technical effect of the present invention, and will not be elaborated further here.

[0048] Preferably, the caffeine is purchased from coffee seed extract of Yangzhou Zhongfu Biotechnology Co., Ltd., and its main active ingredient is caffeine.

[0049] Based on further solutions to the technical problems of the present invention, or simultaneous solutions to multiple technical problems, the preferred solution in the technical solution provided in the first aspect of the present invention includes:

[0050] The first preferred option is a mixture of tetrahydropiperine, caffeine, and leonurine. This technical solution, having already addressed the technical problem of "improving stability," further addresses the technical problem of "further improving stability."

[0051] The second preferred option is that the mass ratio of tetrahydropiperine, caffeine, and leonurine is 1-3:1-3:4-10. This technical solution, based on solving the technical problem of "improving stability," further solves the technical problem of "further improving stability."

[0052] The third preferred option: the oil is selected from at least one of caprylic / capric triglyceride, squalane, egg yolk oil, jojoba seed oil, isononyl isononanoate, and phytosterol / octyldodecyl lauroyl glutamate; the polyol is selected from at least one of glycerol, propylene glycol, and butylene glycol. This technical solution, having solved the technical problem of "improving stability," further solves the technical problem of "further improving stability."

[0053] The fourth preferred option: the oil is selected from at least one of caprylic / capric triglyceride and squalane; the polyol is selected from at least one of glycerol and propylene glycol. This technical solution, based on solving the technical problem of "improving stability", further solves the technical problem of "further improving stability".

[0054] The fifth preferred option, by weight, includes the following raw materials: 1-12 parts alkaloids, 1-8 parts hydrogenated lecithin, 8-28 parts oils, 35-75 parts polyols, and 10-30 parts water. This technical solution, having already addressed the technical problem of "improving stability," further addresses the technical problem of "further improving stability."

[0055] The sixth preferred option, by weight, includes the following raw materials: 1-10 parts alkaloids, 1-5 parts hydrogenated lecithin, 10-25 parts oils, 40-70 parts polyols, and 18-20 parts water. This technical solution, having already addressed the technical problem of "improving stability," further addresses the technical problem of "further improving stability."

[0056] Secondly, the present invention provides a process for preparing the above-mentioned alkaloid liposomes, comprising the following steps:

[0057] S1: Polyol, hydrogenated lecithin and water are mixed to obtain an aqueous phase;

[0058] S2: Oil and alkaloids are mixed to obtain the oil phase;

[0059] S3: Mix the aqueous and oil phases, pre-homogenize, and then homogenize under high pressure to obtain alkaloid liposomes.

[0060] The pre-homogenization process is selected from a shear rate of 10,000-12,000 rpm and a time of 1-3 min.

[0061] Any point value or any range of two point values ​​within the above range can achieve the technical effect of this invention:

[0062] For example, a shear rate of 10000-12000rpm represents shear rates including but not limited to 10000rpm, 11000rpm, 11100rpm, 11200rpm, 11300rpm, 11400rpm, 11500rpm, 11600rpm, 11700rpm, 11800rpm, 11900rpm, and 12000rpm, which will not be elaborated further here.

[0063] If the time is 1-3min, it means that the time is not limited to 1min, 2min, 3min, which will not be elaborated here.

[0064] The preferred pre-homogenization process is a shear rate of 11,000 rpm for 2 minutes.

[0065] The high-pressure homogenization is selected from: pressure of 500-1500 bar, and number of cycles of 5-15.

[0066] Any point value or any range of two point values ​​within the above range can achieve the technical effect of this invention:

[0067] For example, a pressure range of 500-1500 bar represents pressures including but not limited to 500 bar, 550 bar, 600 bar, 650 bar, 700 bar, 750 bar, 800 bar, 850 bar, 900 bar, 950 bar, 1000 bar, 1050 bar, 1100 bar, 1150 bar, 1200 bar, 1250 bar, 1300 bar, 1350 bar, 1400 bar, 1450 bar, and 1500 bar, which will not be elaborated further here.

[0068] For example, a count of 5-15 represents counts including but not limited to 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, and 15 times, which will not be elaborated further here.

[0069] The preferred method for high-pressure homogenization is: a pressure of 1000 bar and 8 cycles.

[0070] Thirdly, the present invention provides the application of the above-described alkaloid liposomes in topical preparations.

[0071] The topical preparation is selected from at least one of the following: creams and serums.

[0072] The present invention has at least the following beneficial effects:

[0073] 1. Compared with the prior art, the present invention has better technical effects in terms of long-term storage particle size stability, temperature cycling particle size stability, dilution stability, centrifugation stability, antibacterial properties, and non-irritating properties.

[0074] 2. Based on the comparison of Examples 1-3 and Comparative Examples 1-5, the present invention utilizes specific combinations of raw materials (especially specific alkaloid combinations) and dosages to achieve new technical effects, providing a safer and more stable alkaloid liposome. The combined technical effect is superior to the sum of the effects of each individual technical means. Attached Figure Description

[0075] Figure 1 Photographs of alkaloid liposomes prepared in Examples 1-3.

[0076] Figure 2 Photographs of alkaloid liposomes prepared for Comparative Examples 1-5.

[0077] Figure 3 Electron microscopy images of the alkaloid liposomes prepared in Example 1.

[0078] Figure 4 Electron microscopy images of the alkaloid liposomes prepared in Example 2.

[0079] Figure 5 Electron microscopy images of the alkaloid liposomes prepared in Example 3.

[0080] Figure 6 Photograph of a sample of the alkaloid liposomes prepared in Example 1 applied to a cream.

[0081] Figure 7 Photograph of a sample of the alkaloid liposomes prepared in Example 1 applied to an essence.

[0082] Figure 8 The images show infrared thermal images of the cream of Example 1 and the control cream before and after massage on the arm; where a, b, c, d and e are infrared thermal images of the cream of Example 1 before massage, 0 min, 8 min, 15 min and 30 min after massage, respectively; f, g, h, i and j are infrared thermal images of the control cream before massage, 0 min, 8 min, 15 min and 30 min after massage, respectively.

[0083] Figure 9 The images show infrared thermal images of the cream of Example 1 and the control cream before and after massage on the body parts; where a and b are infrared thermal images of the cream of Example 1 on the back before massage and 2 hours after massage, respectively; c and d are infrared thermal images of the cream of Example 1 on the right buttock before massage and 2 hours after massage, respectively; e and f are infrared thermal images of the cream of Example 1 on the abdomen before massage and 2 hours after massage, respectively. Detailed Implementation

[0084] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0085] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention are obtained through conventional commercial means.

[0086] The raw material information is shown in Table 1 below:

[0087] Table 1. Raw Material Information

[0088]

[0089] Example 1

[0090] An alkaloid liposome, by weight, is composed of the following raw materials: 60 parts glycerol, 18 parts water, and PHOSPHOLIPON. ® 80H 2 parts, squalane 15 parts, tetrahydropiperine 1 part, caffeine 0.5 parts, leonurine 3.5 parts.

[0091] The preparation process includes the following steps:

[0092] S1: Glycerin, PHOSPHOLIPON ® 80H is mixed with water to obtain an aqueous phase;

[0093] S2: Squalane, tetrahydropiperine, caffeine, and leonurine are mixed to obtain the oil phase;

[0094] S3: The aqueous and oil phases are mixed and pre-homogenized at a shear rate of 11,000 rpm for 2 min, and then homogenized under high pressure at 1000 bar for 8 times to obtain alkaloid liposomes.

[0095] Example 2

[0096] An alkaloid liposome, by weight parts, is composed of the following raw materials: 70 parts propylene glycol, 18 parts water, Phoslip TM HPC 1 part, squalane 10 parts, piperine 0.15 parts, dihydroamarine D 0.15 parts, corydaline 0.7 parts.

[0097] The preparation process includes the following steps:

[0098] S1: Propylene glycol, Phoslip TMHPC is mixed with water to obtain an aqueous phase;

[0099] S2: Squalane, piperine, dihydroamarine alkaloid D and corydaline were mixed to obtain the oil phase;

[0100] S3: The aqueous and oil phases are mixed and pre-homogenized at a shear rate of 10,000 rpm for 3 min, and then homogenized under high pressure at 1500 bar for 5 times to obtain alkaloid liposomes.

[0101] Example 3

[0102] An alkaloid liposome, by weight, is composed of the following raw materials: 40 parts glycerol, 20 parts water, and PHOSPHOLIPON. ® 80H 5 parts, caprylic / capric triglyceride 25 parts, tetrahydropiperine 1.25 parts, caffeine 3.75 parts, leonurine 5 parts.

[0103] The preparation process includes the following steps:

[0104] S1: Glycerin, PHOSPHOLIPON ® 80H is mixed with water to obtain an aqueous phase;

[0105] S2: Caprylic / capric triglyceride, tetrahydropiperine, caffeine and leonurine are mixed to obtain the oil phase;

[0106] S3: The aqueous and oil phases are mixed and pre-homogenized at a shear rate of 12000 rpm for 1 min, and then homogenized under high pressure at 500 bar for 15 times to obtain alkaloid liposomes.

[0107] Comparative Example 1

[0108] An alkaloid liposome, compared to Example 1, except that berberine replaces leonurine, specifically:

[0109] By weight, it consists of the following ingredients: 60 parts glycerin, 18 parts water, and PHOSPHOLIPON. ® 80H 2 parts, squalane 15 parts, tetrahydropiperine 1 part, caffeine 0.5 parts, berberine 3.5 parts.

[0110] The preparation process includes the following steps:

[0111] S1: Glycerin, PHOSPHOLIPON ® 80H is mixed with water to obtain an aqueous phase;

[0112] S2: Squalane, tetrahydropiperine, caffeine, and berberine are mixed to obtain the oil phase;

[0113] S3: The aqueous and oil phases are mixed and pre-homogenized at a shear rate of 11,000 rpm for 2 min, and then homogenized under high pressure at 1000 bar for 8 times to obtain alkaloid liposomes.

[0114] Comparative Example 2

[0115] An alkaloid liposome, compared to Example 1, wherein the alkaloid is tetrahydropiperine, specifically:

[0116] By weight, it consists of the following ingredients: 60 parts glycerin, 18 parts water, and PHOSPHOLIPON. ® 80H 2 parts, squalane 15 parts, tetrahydropiperine 5 parts.

[0117] The preparation process includes the following steps:

[0118] S1: Glycerin, PHOSPHOLIPON ® 80H is mixed with water to obtain an aqueous phase;

[0119] S2: Squalane and tetrahydropiperine are mixed to obtain the oil phase;

[0120] S3: The aqueous and oil phases are mixed and pre-homogenized at a shear rate of 11,000 rpm for 2 min, and then homogenized under high pressure at 1000 bar for 8 times to obtain alkaloid liposomes.

[0121] Comparative Example 3

[0122] An alkaloid liposome, compared to Example 1, except that PHOSPHOLIPON is replaced with LIPOID S75. ® 80H, the remaining raw materials are the same as in Example 1.

[0123] The preparation process includes the following steps:

[0124] S1: Glycerin, LIPOID S75 and water are mixed to obtain an aqueous phase;

[0125] S2: Squalane, tetrahydropiperine, caffeine, and leonurine are mixed to obtain the oil phase;

[0126] S3: The aqueous and oil phases are mixed and pre-homogenized at a shear rate of 11,000 rpm for 2 min, and then homogenized under high pressure at 1000 bar for 8 times to obtain alkaloid liposomes.

[0127] Comparative Example 4

[0128] An alkaloid liposome, compared to Example 1, differs only in the mass ratio of the alkaloids, specifically as follows:

[0129] An alkaloid liposome, by weight, is composed of the following raw materials: 60 parts glycerol, 18 parts water, and PHOSPHOLIPON. ® 80H 2 parts, squalane 15 parts, tetrahydropiperine 4 parts, caffeine 0.5 parts, leonurine 0.5 parts.

[0130] The preparation process is the same as in Example 1.

[0131] Comparative Example 5

[0132] An alkaloid liposome, compared to Example 1, uses hexanediol instead of glycerol and jojoba oil instead of squalane, specifically:

[0133] Based on parts by weight, it consists of the following raw materials: 60 parts hexanediol, 18 parts water, and PHOSPHOLIPON. ® 80H 2 parts, jojoba oil 15 parts, tetrahydropiperine 1 part, caffeine 0.5 parts, leonurine 3.5 parts.

[0134] The preparation process includes the following steps:

[0135] S1: Hexanediol, PHOSPHOLIPON ® 80H is mixed with water to obtain an aqueous phase;

[0136] S2: Jojoba oil, tetrahydropiperine, caffeine, and leonurine are mixed to obtain the oil phase;

[0137] S3: The aqueous and oil phases are mixed and pre-homogenized at a shear rate of 11,000 rpm for 2 min, and then homogenized under high pressure at 1000 bar for 8 times to obtain alkaloid liposomes.

[0138] Test Example 1

[0139] 1. Appearance:

[0140] Figure 1 These are photographs of alkaloid liposomes prepared in Examples 1-3. Figure 2 Photographs of alkaloid liposomes prepared for Comparative Examples 1-5.

[0141] 2. Particle size

[0142] (1) Long-term storage average particle size stability

[0143] Procedure: The alkaloid liposomes prepared in Examples 1-3 and Comparative Examples 1-5 were stored for a long time under ambient light, 45℃, 5℃, -15℃ (protected from light) and ambient light conditions, and particle size was tested on day 0, day 2, day 7, day 14, day 30, day 60 and day 90, respectively.

[0144] The results are shown in Tables 2 and 3 below:

[0145] Table 2. Average particle size (nm)

[0146]

[0147] Table 3. Average particle size (nm)

[0148]

[0149] Note: " / " indicates that the system is not homogeneous and has no testing significance.

[0150] Results: The alkaloid liposomes prepared in Examples 1-3 showed particle size changes of ≤20 nm and <15% after 90 days of storage under light, 45℃, 5℃, -15℃, and room temperature in the dark, indicating stable particle size. The alkaloid liposomes prepared in Comparative Examples 1-5 showed significant particle size changes after 90 days of storage under light, 45℃, 5℃, -15℃, and room temperature in the dark. Specifically, the alkaloid liposomes prepared in Comparative Examples 1, 2, 3, and 5 exhibited stratification or particle precipitation on days 7, 30, 14, and 14, respectively. The alkaloid liposomes prepared in Comparative Example 4 showed a large particle size change under all storage conditions at day 60, increasing by 161-4688 nm, with a change >200%, indicating unstable particle size.

[0151] (2) Temperature cycling particle size stability

[0152] Procedure: The alkaloid liposomes prepared in Examples 1-3 and Comparative Examples 1-5 were stored for a long period of time under temperature cycling conditions of -15℃ to 45℃, and particle size was tested at weeks 0, 1, 2, 3, 4 and 8, respectively.

[0153] The specific temperature cycling conditions are as follows: from the initial temperature (room temperature) to 45℃ (hold for 24 hours) → cool down to -15℃ (hold for 24 hours) → heat up to 45℃ (hold for 24 hours) → cool down to -15℃ (hold for 24 hours) → heat up to 45℃ (hold for 24 hours) → cool down to -15℃ (hold for 24 hours) → return to room temperature (maintain for 24 hours); each cycle lasts for 7 days; each heating and cooling is done in a rapid heating and cooling manner.

[0154] The results are shown in Tables 4 and 5 below:

[0155] Table 4. Stability of Examples

[0156]

[0157] Table 5. Comparative stability

[0158]

[0159] Note: " / " indicates that the system is not homogeneous and has no testing significance.

[0160] Results: The alkaloid liposomes prepared in Examples 1-3, after being stored for 8 weeks under temperature cycling conditions from -15℃ to 45℃, showed a particle size change of ≤15nm and a particle size change range of <15%, indicating that the particle size of the alkaloid liposomes was stable. The alkaloid liposomes prepared in Comparative Examples 1-5, after being stored for 8 weeks under temperature cycling conditions from -15℃ to 45℃, showed stratification in Comparative Examples 1, 3, and 4 at weeks 2, 1, and 4, respectively. The alkaloid liposomes prepared in Comparative Examples 2 and 5 showed a particle size increase of 1747nm and 1458nm at weeks 2 and 4, respectively, an increase of >10 times, indicating that the particle size was unstable.

[0161] (3) Dilution stability

[0162] Procedure: The alkaloid liposomes prepared in Examples 1-3 and Comparative Examples 1-5 were diluted with water by 2, 5, 10, 20, 50, 100, and 200 times, respectively, and allowed to stand. The particle size was tested on day 0, day 3, day 7, and day 14.

[0163] The results are shown in Tables 6 and 7:

[0164] Table 6. Average particle size (nm)

[0165]

[0166] Table 7. Average particle size (nm)

[0167]

[0168] Results: The alkaloid liposomes prepared in Examples 1-3, after being diluted 2-200 times and stored for 14 days, showed a particle size change of less than 10 nm, with a change range of <10%, indicating that the particle size of the alkaloid liposomes was stable. The alkaloid liposomes prepared in Comparative Examples 1-5, after being diluted 2-200 times and stored for 14 days, showed stratification in Comparative Examples 1, 3, and 5 on day 14. The particle sizes of the alkaloid liposomes prepared in Comparative Examples 2 and 4 increased by 398 nm and 66 nm, respectively, with an increase of >45%, indicating that the particle size was unstable.

[0169] (4) Centrifugal stability

[0170] Procedure: The alkaloid liposomes prepared in Examples 1-3 and Comparative Examples 1-5 were centrifuged at 3000 rpm for 30 min, and the sample state and particle size changes were observed.

[0171] The results are shown in Table 8:

[0172] Table 8. Centrifugal Stability

[0173]

[0174] Results: The alkaloid liposomes prepared in Examples 1-3 showed a particle size change of less than 3 nm before and after centrifugation, with a change range of <1.5%, indicating that the alkaloid liposomes were stable after centrifugation. After centrifugation, the alkaloid liposomes prepared in Comparative Examples 1-5 showed stratification. The alkaloid liposomes prepared in Comparative Example 4 showed a particle size increase of 332 nm, with an increase of >240%, indicating that the particle size was unstable.

[0175] Test Example 2

[0176] Microbiological testing: The alkaloid liposomes prepared in Examples 1-3 and Comparative Examples 1-5 were subjected to microbiological testing. The total number of colonies, thermotolerant Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, molds and yeasts were tested in accordance with the microbiological testing methods in the "Cosmetic Safety Technical Specifications (2015 Edition)". The results are summarized in Table 9 below.

[0177] Table 9. Microbial Detection Results

[0178]

[0179] Results: Microbiological testing showed that the total bacterial count, mold count, and yeast count in the alkaloid liposomes of Examples 1-3, Comparative Examples 3 and 5 met the requirements, and thermostable Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa were not detected, indicating that the microbiological testing was qualified. However, the Staphylococcus aureus count and total bacterial count in Comparative Examples 1, 2, and 4 did not meet the requirements.

[0180] Test Example 3

[0181] Electron microscopy characterization was performed on the alkaloid liposomes prepared in Examples 1-3:

[0182] Figure 3 Electron microscopy images of the alkaloid liposomes prepared in Example 1. Figure 4 Electron microscopy images of the alkaloid liposomes prepared in Example 2. Figure 5 Electron microscopy images of the alkaloid liposomes prepared in Example 3.

[0183] It can be seen that the alkaloid liposomes prepared in Examples 1 and 3 are white to yellow translucent liquids, while the alkaloid liposomes prepared in Example 2 are brown translucent liquids. The alkaloid liposomes prepared in Examples 1, 2, and 3 are spherical particles with a round and full appearance and an average particle size between 100-200 nm. It can be seen that the alkaloid liposomes prepared in the examples of this invention have uniform and stable particle size, and further irritation tests were conducted.

[0184] Test Example 4

[0185] Stimulation test

[0186] sample:

[0187] (1) Alkaloid liposomes prepared in Examples 1-3 (diluted with water to 4%, w / w)

[0188] (2) Active substances of the same concentration:

[0189] Mixture 1 (a squalane dispersion stock solution of 1% tetrahydropiperine + 0.5% caffeine + 3.5% leonine, further diluted with squalane to 4%, w / w);

[0190] Mixture 2 (a squalane dispersion stock solution of 0.15% piperine + 0.15% dihydroazoline D + 0.7% corydaline, further diluted with squalane to 4%, w / w);

[0191] Mixture 3 (a stock solution of caprylic / capric triglyceride dispersion of 1.25% tetrahydropiperine + 3.75% caffeine + 5% leonine, further diluted with caprylic / capric triglyceride to 4%, w / w).

[0192] Human skin patch tests were conducted separately, referring to the method of closed patch test in the "Cosmetic Safety Technical Specifications (2015 Edition)". Specifically, qualified patch testing equipment was selected, and 0.020 mL of the test substance was placed in the patch testing device using the closed patch test method. The patch was then applied to the inside of the subject's arm with hypoallergenic adhesive tape. The test substance was removed after 24 hours, and skin reactions were observed at 0.5 h, 24 h, and 48 h after removal. The results were recorded according to the skin reaction grading standard in the "Cosmetic Safety Technical Specifications (2015 Edition)" (as shown in Table 10 below). The results are shown in Table 11.

[0193] Subject information: 12 healthy Chinese male / female subjects, aged 22-45 years.

[0194] Table 10. Skin Reaction Rating Table

[0195]

[0196] Table 11. Summary Results of Cosmetic Human Skin Patch Tests

[0197]

[0198] Results: Human skin patch tests showed no skin reactions in Examples 1-3. Mixtures 1, 2, and 3 resulted in 9 cases of Grade 1 skin reactions, 5 cases of Grade 2 skin reactions, and 1 case of Grade 3 skin reaction. Specifically, Mixture 1 showed 3 cases of Grade 1 skin reactions and 2 cases of Grade 2 skin reactions; Mixture 2 showed 2 cases of Grade 1 skin reactions and 1 case of Grade 2 skin reaction; and Mixture 3 showed 4 cases of Grade 1 skin reactions, 2 cases of Grade 2 skin reactions, and 1 case of Grade 3 skin reaction. This indicates that the alkaloid liposomes of Examples 1-3 have good safety profiles, and liposome encapsulation helps reduce the irritation of the alkaloids themselves.

[0199] Application Example 1

[0200] 1. Cream

[0201] Experimental design: 1%-4% (w / w) of alkaloid liposomes prepared in Example 1 were added to the cream, with no alkaloid liposomes added as a control group, resulting in an alkaloid liposome cream; its formulation is shown in Table 12:

[0202] Table 12. Formula

[0203]

[0204] Preparation process: Phase A and Phase B were heated to 80℃ respectively, mixed, and homogenized at 13000 rpm for 2 min; the mixture was then stirred and cooled to 50℃, and Phases C and D were added sequentially, mixed evenly, and discharged. The physicochemical properties of the resulting cream are shown in Table 13.

[0205] Table 13. Physicochemical properties of creams

[0206]

[0207] Results: Adding 1%-4% of alkaloid liposomes to creams did not change their appearance and had little effect on viscosity and pH, indicating that alkaloid liposomes can be used in creams.

[0208] 2. Serum

[0209] Experimental design: 1%-4% (w / w) of alkaloid liposomes prepared in Example 1 were added to the essence, with no alkaloid liposomes added as a blank control, resulting in an alkaloid liposome essence; its formulation is shown in Table 14:

[0210] Table 14. Formula

[0211]

[0212] Preparation process: Phase A was heated to 80℃ and homogenized for 2 minutes; then cooled to 50℃ with stirring, and phases B, C, D, and E were added sequentially, mixed evenly, and discharged. The physicochemical properties of the obtained essence are shown in Table 15.

[0213] Table 15. Physicochemical Properties of Serums

[0214]

[0215] Results: Adding 1%-4% alkaloid liposomes to serums had little effect on viscosity and pH. The addition of alkaloid liposomes changed the original appearance, making it appear as a white, semi-transparent substance with a bluish sheen. This indicates that alkaloid liposomes can be used in serums to improve their appearance.

[0216] Application Example 2

[0217] Experimental Design:

[0218] Subject profile: Ten healthy Chinese male / female subjects aged 18-60 years with poor skin barrier and dryness of the hands were selected.

[0219] Instructions for use: After cleansing the skin, take an appropriate amount of the product (i.e., the 4% (w / w) alkaloid liposome cream prepared in Example 1) and the control cream (containing the same net content of unencapsulated alkaloids) and apply it to both arms, once in the morning and once in the evening.

[0220] Testing methods: A before-and-after control method was used. Before using the test product, and after using the product for 14 and 28 consecutive days, the skin moisture content was measured using a Corneometer device, and the transepidermal water loss was measured using a Tewameter device.

[0221] 14-day improvement rate = (D14-day test value - D0 baseline value) / D0 baseline value × 100%.

[0222] 28-day improvement rate = (D28-day test value - D0 baseline value) / D0 baseline value × 100%.

[0223] Table 16. Results of Skin Moisture Content and Transepidermal Water Loss Tests

[0224]

[0225] Note: The lower the transepidermal water loss value, the better the skin barrier function.

[0226] Results: After 14 and 28 days of using the cream of Example 1, skin moisture content increased by 14.24% and 26.82%, respectively. After 14 and 28 days of using the control cream, skin moisture content increased by 9.30% and 7.32%, respectively. This demonstrates that the alkaloid liposomes prepared in Example 1 have a better effect on improving skin moisture content when applied to the cream. After 14 and 28 days of using the cream of Example 1, transepidermal water loss decreased by 13.14% and 26.47%, respectively. After 14 and 28 days of using the control cream, transepidermal water loss decreased by 2.34% and -4.58%, respectively. This demonstrates that the alkaloid liposomes prepared in Example 1 can more effectively reduce skin moisture loss and have a better skin barrier repair effect when applied to the cream.

[0227] Application Example 3

[0228] Experimental Design:

[0229] Subject information: Twelve healthy Chinese male / female subjects with normal skin condition, aged 22-55 years, were selected.

[0230] Instructions for use: Apply 3g of the 4% (w / w) alkaloid liposome cream prepared in Example 1 to one arm of the subject, and apply 3g of the control cream (containing the same net amount of unencapsulated alkaloids) to the other arm. Massage both arms for 5 minutes.

[0231] Testing Method: A before-and-after control method was used. Images were captured using an infrared thermal imager (KIR-2008P, Wuhan Huajingkang Optoelectronics) before massage and at 0 min, 8 min, 15 min, and 30 min after massage. Simultaneously, feedback from subjects was collected (skin absorption speed, discomfort intensity, and overall satisfaction; evaluation indicators and scoring methods are shown in Table 17). The average values ​​were used to obtain the comparative test results. Results are shown in Table 18 and... Figure 8 .

[0232] Table 17. Evaluation Indicators and Scoring Methods

[0233]

[0234] Table 18. Comparison Test Results

[0235]

[0236] Results: Infrared thermal imaging tests and participant feedback showed that the cream of Example 1 significantly accelerated local blood circulation, increased local temperature over a wider area more quickly, and the warming sensation lasted for over 30 minutes. Furthermore, participants reported that the cream containing Example 1 absorbed quickly, experienced minimal discomfort during use, and was generally satisfied. The comparison cream, however, had a weaker effect on promoting blood circulation and increasing local temperature, with a smaller temperature increase area and a warming sensation lasting only about 15 minutes. In terms of skin feel and absorption speed, the comparison cream absorbed more slowly, causing skin discomfort during use, resulting in lower overall satisfaction.

[0237] This test further examined the effects of the cream from Example 1 on other parts of the body. Images were acquired and analyzed using an infrared thermal imager (KIR-2008P, Wuhan Huajingkang Optoelectronics). The results showed that the cream effectively promoted microcirculation and increased local temperature. (See results below.) Figure 9 .

[0238] The alkaloid liposomes prepared in Example 1 demonstrate that when applied to a cream, they can be rapidly absorbed through the skin, thereby promoting local blood circulation, increasing local temperature, reducing skin irritation from the alkaloids, and resulting in a better user experience. Simultaneously, the application of liposomes enhances the sustained-release effect of the alkaloids, thus prolonging the state of improved local microcirculation and temperature.

[0239] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. An alkaloid liposome, characterized in that, By weight, it includes the following raw materials: The composition comprises 1-12 parts alkaloids, 1-8 parts hydrogenated lecithin, 8-28 parts oils, 35-75 parts polyols, and 10-30 parts water; wherein the alkaloids are tetrahydropiperine, caffeine, and leonine in a mass ratio of 1-3:1-3:4-10; the hydrogenated lecithin is selected from at least one of PHOSPHOLIPON 80H or Phoslip HPC; the oils are selected from caprylic / capric triglycerides or squalane; and the polyol is glycerol.

2. An alkaloid liposome, characterized in that, The product is composed of the following ingredients by weight: 70 parts propylene glycol, 18 parts water, 1 part Phoslip HPC, 10 parts squalane, 0.15 parts piperine, 0.15 parts dihydroamarine alkaloid D, and 0.7 parts corydaline.

3. The alkaloid liposome of claim 1, wherein, By weight, it includes the following ingredients: 1-10 parts alkaloids, 1-5 parts hydrogenated lecithin, 10-25 parts oils, 40-70 parts polyols, and 18-20 parts water.

4. The process for the preparation of liposomes of alkaloids according to claim 1 or 3, characterized in that, Includes the following steps: S1: Polyol, hydrogenated lecithin and water are mixed to obtain an aqueous phase; S2: Oil and alkaloids are mixed to obtain the oil phase; S3: Mix the aqueous and oil phases, pre-homogenize, and then homogenize under high pressure to obtain alkaloid liposomes.

5. The manufacturing process of claim 4, wherein, In step S3, the pre-homogenization shear rate is 10000-12000 rpm, and the time is 1-3 min; the high-pressure homogenization pressure is 500-1500 bar, and the number of times is 5-15.

6. The use of the alkaloid liposomes according to any one of claims 1-3 in the preparation of topical formulations.

Images