A Stable Ion-Resistant Gel Matrix Composition and Its Use

Abstract

The present invention belongs to the fields of medicine and chemistry, and relates to a stable ionic-resistant gel matrix composition and its uses. Specifically, it relates to a novel material prepared from cationized Bletilla striata polysaccharide (CBSP) and carbomer (CP), especially a novel gel matrix that is stable, ionic-resistant and has good rheological properties, as well as its uses as a gel matrix in the fields of medicine and chemistry. The present invention makes up for the deficiencies of existing gel matrix materials and has the characteristic of enhancing the stability of the gel system; the new matrix material exhibits excellent pharmaceutical excipient characteristics, especially in the application of preparing gels. It can provide key basic materials for the development of gel preparations with high stability and good rheological properties, especially for the development of highly stable gel preparations with complex components and containing ions.

Description

Technical Field

[0001] The present invention belongs to the fields of medicine and chemistry, and relates to a stable ionic-resistant gel matrix composition and its uses. Specifically, it relates to a novel material prepared from cationized Bletilla striata polysaccharide (CBsP) and carbomer (CP), especially a novel gel matrix that is stable, ionic-resistant and has good rheological properties, as well as its uses as a gel matrix in the fields of medicine and chemistry. Background of the Invention

[0002] Gels refer to thick liquids or semi-solid preparations with gel characteristics prepared from the raw material drug and excipients that can form gels (also known as gel matrices). According to the different gel matrices, gels are divided into aqueous gels and oily gels. Aqueous gels have the characteristics of easy spreading, good comfort, no greasy feeling, easy to wash off, can absorb tissue exudate, do not interfere with the normal physiological functions of the skin, have a certain water retention effect, and can promote the transdermal absorption of active ingredients, etc., and are widely used in the fields of medicine, daily chemicals, etc., and are deeply favored by patients and consumers.

[0003] The composition of gels generally includes active components, gel matrices, pH regulators, and additional agents such as humectants, preservatives, transdermal absorption promoters, antioxidants, flavors, etc. can also be added according to needs. Among them, the gel matrix, as the excipient of the gel, is closely related to the forming state of the gel, the stability of the system, rheological properties, skin feel and other usage properties, as well as the storage and transportation of the preparation. The quality of the matrix itself largely determines the success or failure of the development of gel preparation products and the quality of the products.

[0004] Carbomer, also known as carbopol, alias: polyacrylic acid, carboxyvinyl copolymer, is a high molecular polymer cross-linked by acrylic acid and allyl sucrose or allyl pentaerythritol. It is the most commonly used matrix for aqueous gels, has important uses such as thickening and suspending, has a simple preparation process, good rheological properties, and no irritation to the skin and mucous membranes, etc., and is widely used in emulsions, creams, gels. It is a very excellent gel matrix material. Although the process of preparing gels with carbomer is simple and can endow the gels with good rheological properties, its disadvantage is that carbomer has a relatively strong sensitivity to the gel environment, which is manifested in that the presence of ions will significantly reduce its thickening efficiency; trace amounts of iron or other transition metals will cause irreversible viscosity decrease of the gel, seriously damaging the stability of the system; in addition, high shear, ultraviolet irradiation or persistent stirring will also cause viscosity loss of carbomer gels, reducing the gel viscosity. The sensitivity of carbomer, especially ionic sensitivity, is the main problem in its application. Acids, mineral ions cannot be added to gels based on carbomer, and certain bacteriostatic agents need to be avoided or can only be used at low concentrations, otherwise the consistency will decrease sharply or even disappear. In order to improve the ionic resistance of carbomer matrices, the United States The company has developed improved gel matrices, namely Carbopol 1342 and Carbopol ETD 2020. However, the above-mentioned Carbopol still has a relatively low ion tolerance. Especially when divalent or trivalent ions exist in the system, not only will the viscosity of the system decrease irreversibly, but also insoluble precipitates may be generated. As a gel matrix material with excellent rheological properties, the lack of stability of matrix properties, especially ion tolerance, limits the application of Carbopol matrices, and may potentially have an adverse impact on the quality of gel products containing ionic components or multiple components, such as complex traditional Chinese medicine extracts.

[0005] Currently, for gel systems containing trace ions, methods such as adding metal ion chelating agents or antioxidant stabilizers are mostly used to improve the stability of the ionic system. For example, by adding EDTA-2Na to chelate metal ions in the system or maintain the stability of the system. However, if the dosage is excessive, it is likely to cause some skin problems, especially for sensitive skin types; the complexing effect of EDTA-2Na is also affected by the acidity of the solution, and the effective complexing concentration decreases as the acidity of the solution increases; some studies have found that in addition to causing changes in the function and morphology of cell membranes, EDTA-2Na may also induce DNA damage in corneal epithelial cells at low concentrations (0.0001 - 0.01%); for the case of ionic active ingredients, it is necessary to consider the complexation of EDTA-2Na with ions, which may affect the effectiveness of the active ingredients.

[0006] In addition, in response to the ion sensitivity problem, the French company Seppic has also successively developed a gel matrix polyacrylate cross-linked polymer-6 (zen) with ion tolerance characteristics. However, its thickening property is not ideal and it still needs to be used in combination with other matrices.

[0007] The forming state and skin feel of the gel are related to the structure and rheological properties of the matrix, and the rheological properties are also related to the quality of the preparation, usage performance, etc. Therefore, in addition to the durable stability of the matrix such as ion tolerance and shear resistance, controlling the rheological parameters of the gel is also the key to determining the stability, feel in use and preparation performance of the gel. An ideal gel should also have the following rheological characteristics:

[0008] (1) It should have an appropriate viscosity. The greater the viscosity, the lower the movement rate of the particles in the system, and it is not easy to show stratification. However, at the same time, the required fluidity of the system, the ease of application and skin comfort should be taken into account;

[0009] (2) Appropriate pseudoplastic rheological characteristics, that is, the phenomenon of shear thinning. When preparing, the viscosity of the gel decreases under the action of mechanical stirring, which is conducive to the uniform dispersion of each component;

[0010] (3) Coupled with a certain thixotropy, it can ensure that the gel has appropriate spreadability during application, and can ensure the gel form when standing still;

[0011] (4) Appropriate yield value, i.e., shear resistance, affects the suspension performance and stability performance of the preparation. During the preparation design, the yield stress of the gel needs to meet the requirements that it is easy to flow when extruded, easy to spread when applied, and can overcome its own gravity when not stressed, without damaging the network structure of the system, maintaining the semi-solid state, and maintaining the suspension ability of the system, so as to ensure the stability of the gel and prevent the gel state from being damaged during use, carrying, and transportation.

[0012] As a semi-solid gel preparation that is convenient and comfortable to use but thermodynamically unstable, good rheological properties and relatively better stability should be the unremitting pursuit of this preparation. However, under the existing conditions of carbomer matrix materials, ionic drugs, plant extracts, ionic surfactants, complex components of traditional Chinese medicine compound drugs, and certain preservatives added in the gel preparation formula will all cause instability phenomena, and even lead to problems such as discoloration, delamination, slight or irreversible decrease in the system viscosity, and poor fluidity of the product during the validity period, affecting its use performance. Therefore, developing gel matrix materials with better ion resistance, shear resistance and other durable stabilities and good rheological properties has positive significance for improving the performance and quality stability of gel products and meeting the development of diversified gel products.

[0013] Some English abbreviations that may be involved in the present invention and their corresponding Chinese names:

[0014]

[0015] Summary of the Invention

[0016] The present invention provides a novel modified polyacrylic acid auxiliary material, specifically a novel gel matrix prepared from cationized Bletilla striata polysaccharide (abbreviated as CBSP) and an anionic polymer, especially a new gel matrix with better stability, ion resistance, resistance to complex components of traditional Chinese medicine and good rheological properties. The new matrix material of the present invention exhibits excellent pharmaceutical auxiliary material characteristics and can provide key basic materials for the development of gel preparations with complex components and high stability containing ionic components.

[0017] The present invention cationizes Bletilla striata polysaccharide. On this basis, cationized Bletilla striata polysaccharide CBSP and an anionic polymer are combined to construct a novel gel matrix composition material by using the electrostatic interaction between the two. Among them, the anionic polymer can be polyacrylic acid, such as carbomer (abbreviated as CP).

[0018] A CBSP-CP composition, characterized in that it includes CBSP and CP that can form a composition with CBSP.

[0019] Further, in the composition, CP is selected from Carbopol 940, Carbopol 1342, Carbopol ETD2020, etc., and Carbopol ETD 2020 is further preferred. That is, CBSP and CP form a composition (CBSP-CP), especially CBSP and Carbopol ETD 2020 form a composition.

[0020] A CBSP-CP composition, wherein the ratio range of CBSP to carbomer (CP) is (1-3):(1-3) by mass; further, the preferred ratio range of the two is 2:1.

[0021] The novel ion-resistant gel matrix (CBSP-CP) provided by the present invention, that is, the novel gel matrix composition, is characterized in that it is composed of the above-mentioned cationized Bletilla striata polysaccharide CBSP and the anionic polymer carbomer CP. Its preparation process:

[0022] Disperse carbomer in water, stir and dissolve, and adjust the pH to neutral to obtain phase A;

[0023] Disperse the cationized Bletilla striata polysaccharide in water, stir and dissolve to obtain phase B;

[0024] Slowly add phase B to phase A to obtain it; or dry and store for later use.

[0025] The present invention also discloses a cationized Bletilla striata polysaccharide (CBSP), which is characterized in that: it has characteristic peaks of quaternary ammonium salt in its infrared spectrum, such as the absorption peak of C-N bond on the quaternary ammonium salt (1670 cm -1 , TENSOR 27 type infrared spectrometer), the stretching vibration peak of C-H bond on the quaternary ammonium salt (1568 cm -1 , TENSOR 27 type infrared spectrometer).

[0026] The cationized Bletilla striata polysaccharide (CBSP) described in the present invention can be used to carry out cationization modification on Bletilla striata polysaccharide (BSP) by the Williamson synthesis method, and the reaction mechanism is as follows:

[0027] 1) Alkylation reaction:

[0028]

[0029] Among them, R1 is selected from H, Na;

[0030] 2) Etherification reaction:

[0031]

[0032] Among them, R2 is selected from H,

[0033] Specifically, the following method can be adopted. Using urea solution as the dispersion medium, 3-chloro-2-hydroxypropyltrimethylammonium chloride (CTA) as the cationic etherifying agent, and sodium hydroxide (NaOH) as the catalyst, the modification process was investigated and studied through single-factor screening conditions and CCD-RSM optimization test method. Combining with the actual preparation process, the conditions of the modification process were formulated as follows: etherification temperature 50-60 °C, etherification time 5 h-6 h, alkali-ether molar ratio 2:1-1:2;

[0034] Furthermore, the etherification temperature of 55 °C, the etherification time of 5.5 h, and the molar ratio of alkali to ether of 1:1 are preferred.

[0035] The cationic products of Bletilla striata polysaccharide were characterized by infrared spectroscopy, nuclear magnetic resonance and thermogravimetric analysis. The results showed that BSP exhibited cationic characteristics under the above conditions and could form cationic Bletilla striata polysaccharide (CBSP). Compared with BSP, the transparency of the CBSP solution was significantly improved and the water solubility was increased; the Zeta potential measurement results showed that BSP was almost uncharged when the solution pH was less than 7, and when the pH was greater than 7, the solution carried a certain amount of negative charge, while CBSP carried a certain amount of positive charge under different pH conditions, and within the range of pH 5-7, its potential value only changed slightly, which provided a guarantee for binding with negatively charged anionic polymers through electrostatic interaction.

[0036] The acrylic polymer CP has good thickening property, transparency and rheology, but its stability is relatively poor. Especially the addition of substances such as ions and salts will seriously damage the stability of the system. As a gel matrix material with excellent rheological properties, the lack of stability of the matrix properties, especially the ion resistance, limits the application of Carbopol matrix.

[0037] In the present invention, BSP was first cationized to obtain CBSP, and then combined with anionic polymer CP to prepare a CBSP-CP composition, aiming to provide a performance excellent ion-resistant gel matrix composition material with good thickening property, rheology and good ion resistance at the same time.

[0038] Testing of the matrix properties of the ion-resistant gel matrix composition CBSP-CP of the present invention. Taking three matrix materials, namely traditional Carbopol940 (CP 940), modified (ion-resistant modified) Carbopol ETD 2020 (CP2020), and polyacrylate cross-linked polymer-6 (Zen), as references, the present invention evaluated the matrix properties such as ion stability, rheological properties, and biocompatibility of the CBSP-CP gel matrix composition. The results showed that: ① Ion resistance. The matrix of the CBSP-CP composition of the present invention showed very good ion resistance. Using different concentrations of NaCl and CaCl2 as Na + 、Ca2+ For the ionic reagents, the traditional CP940, and the improved (ion-resistant improved) CP 2020, with the addition of ions, the viscosity shows a significant downward trend. When the mass fraction of NaCl is 0.5% and the mass fraction of CaCl2 is 0.3%, the viscosity of the system almost drops to 0; the viscosity reduction of Zen is slightly slower than that of CP 940 and CP 2020, but when Ca 2+ is 0.3%, the viscosity also almost completely disappears. Under the same conditions, the viscosity of CBSP-CP almost remains unchanged with the increase of Na + , Ca 2+ concentration. Even in the presence of high-concentration ions such as 1.5% NaCl and 1.0% CaCl2, the viscosity remains almost stable, and the viscosity retention rate is still greater than 90%, showing that the CBSP-CP composition matrix has excellent ion resistance. ② Rheological properties. CBSP-CP has a similar thickening property to the other three commercially available matrices, CP 940, CP 2020, and Zen. They all belong to non-Newtonian pseudoplastic fluids. The rheological behavior of CBSP-CP is similar to that of CP 940 and has a certain thixotropy, with good rheological properties, which is beneficial to the extrusion, spreading, and stability of gel preparations. ③ Biocompatibility. Biocompatibility evaluation is often used to study the interaction between polymer materials and organisms. The biocompatibility of the CBSP-CP composition matrix of the present invention was investigated by using blood compatibility and skin safety tests respectively. The results of the erythrocyte hemolysis experiment show that the hemolysis rates of CBSP-CP at different concentrations (1, 5, 10, 20 mg / mL) are all lower than the limit value of 5%, indicating good biocompatibility of CBSP-CP; the acute skin irritation experiment on rats shows that there are no reactions such as erythema and edema after administration of the CBSP-CP composition of the present invention, indicating good skin safety of the CBSP-CP composition matrix.

[0039] Evaluation of the application results of the CBSP-CP ion-resistant gel matrix composition of the present invention. The forming state, skin feel, stability, etc. of the gel are closely related to the performance of the matrix used. Especially for gels with complex preparation components and ionic components, the performance of the matrix largely determines the success or failure and quality of the gel product preparation. In the present invention, the extracts of compound traditional Chinese medicines dandelion and scutellaria baicalensis and zinc gluconate are used as model components of complex components and metal ions. Compound traditional Chinese medicine puhuang zinc gel is prepared with three different types of matrices, namely traditional Carbopol 940, modified Carbopol ETD 2020, and polyacrylate cross-linked polymer-6 (Zen), and the CBSP-CP composition of the present invention as the matrix, and the application effect of the CBSP-CP matrix composition in the preparation of the gel is compared. The research results show that: ① Stability. Compared with the other three matrices, the compound puhuang-zinc gel prepared with the CBSP-CP of the present invention as the matrix has better stability in terms of the appearance properties, viscosity, and content of active index components of the gel under high temperature (40 °C, 60 °C), low temperature (-4 °C), and strong light (4500 Lx) conditions (P < 0.01). It shows that the CBSP-CP of the present invention can improve the stability of gels, especially gels containing complex components and metal ion components. ② Rheological properties. When the compound puhuang-zinc gel containing complex components of metal ions and traditional Chinese medicine extracts is prepared with the CBSP-CP of the present invention, the decrease in the steady-state apparent viscosity is the smallest, indicating that CBSP-CP has a better viscosity-increasing effect and a role in maintaining the viscosity of the system for complex-component gels. In addition, the rheological curves of the compound puhuang-zinc gel prepared with different matrix materials show that when CBSP-CP is applied to ionic and complex-component systems, it shows pseudoplastic fluidity, which is beneficial for spreading, and at the same time shows better shear resistance compared with the other three matrices. ③ Transdermal permeation characteristics. Taking baicalin as the index component, the 12-hour cumulative permeation rate of baicalin in the compound puhuang zinc gel samples prepared with each matrix is the highest with CBSP-CP, reaching 73.31 ± 2.84%, and the lowest with CP 940, which is 49.12 ± 2.87%. At the same time, the 12-hour skin retention amount of baicalin in the compound gel sample with CBSP-CP as the matrix is also relatively large, which is beneficial for forming a drug reservoir on the skin and playing a long-term effect. ④ Moisturizing effect. The measurement results of skin water content and transepidermal water loss show that compared with the three commercially available matrices, the skin water content (MMV) of the compound gel with CBSP-CP as the matrix is significantly increased at each time point (P < 0.05); the transepidermal water loss (TEWL) is significantly decreased (P < 0.01), indicating that the CBSP-CP composition of the present invention as a gel matrix helps to improve the skin barrier function and enhance the moisturizing effect.

[0040] The CBSP-CP composition of the present invention, namely a novel ion-resistant gel matrix composition, makes up for the shortcomings of existing pharmaceutical excipients, has the characteristics of synergistic viscosity increase and enhanced gel system stability, and the new matrix material exhibits excellent pharmaceutical excipient properties, especially in the preparation of gels.

[0041] The CBSP-CP composition provided by the present invention is used in the preparation of pharmaceuticals and daily chemicals; it can also be used to prepare gels, especially as a gel matrix.

[0042] Furthermore, the present invention uses this novel ion-resistant gel matrix composition as the matrix of a compound gel with dandelion, Scutellaria baicalensis extract (abbreviated as "Pu-Huang extract") and zinc gluconate as the main drugs to prepare the compound traditional Chinese medicine Pu-Huang-Zinc gel. Through the applied research on the stability, rheological properties and skin adaptability of compound gels with different matrices, the comparative differences of different matrices in the application of gels are obtained, and the application of this novel matrix in the preparation of complex component gel preparations with excellent and stable use performance is realized. Furthermore, it can provide key basic materials for the development of more types of complex component gel preparations with excellent and stable performance. Description of the Drawings

[0043] Figure 1 : Infrared spectra of cationized Bletilla striata polysaccharide (CBSP) and Bletilla striata polysaccharide (BSP);

[0044] Figure 2 : 1H nuclear magnetic resonance spectrum of CBSP;

[0045] Figure 3 : TG and DTG curves of Bletilla striata polysaccharide (a) and cationized Bletilla striata polysaccharide (b);

[0046] Figure 4 : Zeta potentials of cationic Bletilla striata polysaccharide (CBSP) and Bletilla striata polysaccharide (BSP) at different pH values;

[0047] Figure 5 : Appearance properties of CBSP, BSP, and CP2020 solutions;

[0048] Figure 6 : Transmittance of CBSP and BSP with different mass fractions;

[0049] Figure 7 : SEM images of CBSP, CP, and CBSP-CP at 120X (600X) are represented by A (a), B (b), and C (c) respectively;

[0050] Figure 8 : (a), (b) CBSP-CP and commercially available matrix materials under different ionic conditions (Na + , Ca2+ ) viscosity change curve;

[0051] Figure 9 : shear stress and viscosity vs. shear rate curves;

[0052] Figure 10 : shear stress vs. shear rate curve;

[0053] Figure 11 : blood compatibility of different matrix materials;

[0054] Figure 12 : hemolysis rates of different matrix materials at different concentrations;

[0055] Figure 13 : Appearance pictures of the back skin of rats smeared with different matrix gels once, where: a, CBSP; b, CP 940; c, CP 2020; d, Zen; e, CBSP-CP;

[0056] Figure 14 : γ-η rheological curves of CP-CBSP and commercially available matrix compound Puhuang-Xin gels;

[0057] Figure 15 : percutaneous penetration curves of CP-CBSP and commercially available matrix Puhuang-Xin gels

[0058] Figure 16 : skin retention rate results of CP-CBSP and commercially available matrix Puhuang-Xin gels

[0059] Figure 17 : effects of CP-CBSP and commercially available matrix on skin water content of Puhuang-Xin gel (n = 6);

[0060] Figure 18 : effects of CP-CBSP and commercially available matrix on skin water loss of Puhuang-Xin gel (n = 6). Detailed implementation manners

[0061] Bletilla striata polysaccharide (BSP, provided by Xi'an Parnir Biotechnology Co., Ltd., Shaanxi, China); Carbomers (CP): Carbopol 940 (CP940, provided by Lubrizol, USA), Carbopol 1342 (CP1342, provided by Lubrizol, USA), Carbopol ETD 2020 (CP 2020, provided by Lubrizol, USA), Carbopol Ultrez20 (CP U20, provided by Lubrizol, USA), polyacrylate cross - polymer - 6 (zen, provided by Seppic, France).

[0062] Example 1 Preparation of cationic Bletilla striata polysaccharide

[0063] Dissolve BSP in NaOH / urea solution to obtain a homogeneous system. Then add NaOH to alkalize at room temperature, raise the temperature, add NaOH to the system and slowly dropwise add the etherifying agent 3 - chloro - 2 - hydroxypropyltrimethylammonium chloride (CTA). The molar ratio of base to ether in the system is 1:2. Carry out etherification in a constant - temperature water bath for 5 h, adjust the pH to neutral, and the etherification reaction ends. Subsequently, place it in an activated dialysis bag with a cut - off molecular weight of 8 - 14 KD, use ultrapure water as the permeation medium for purification, and change the exudate every 3 h; after purification, filter by suction, rotary evaporation, freeze - dry, and pulverize to obtain cationic Bletilla striata polysaccharide (CBSP).

[0064] Example 2 Preparation of cationic Bletilla striata polysaccharide

[0065] Dissolve BSP in NaOH / urea solution to obtain a homogeneous system. Then add NaOH to alkalize at room temperature for 10 min, then raise the temperature to 55 °C, add NaOH to the system and slowly dropwise add the etherifying agent 3 - chloro - 2 - hydroxypropyltrimethylammonium chloride (CTA). The molar ratio of base to ether in the system is 2:1. Carry out etherification in a constant - temperature water bath for 6 h, adjust the pH to neutral, and the etherification reaction ends. Subsequently, place it in an activated dialysis bag with a cut - off molecular weight of 8 - 14 KD, use ultrapure water as the permeation medium for purification, and change the exudate every 3 h; after purification, filter by suction, rotary evaporation, freeze - dry for 48 h to obtain cationic Bletilla striata polysaccharide (CBSP).

[0066] Example 3 Preparation of cationic Bletilla striata polysaccharide

[0067] Dissolve BSP in the NaOH / urea solution to obtain a homogeneous system. Then add part of NaOH and alkalize at room temperature for 10 min. After that, raise the temperature to 54 °C, add the remaining NaOH to the system and slowly dropwise add the etherifying agent CTA. The molar ratio of alkali to ether in the system is 1:1. Carry out etherification in a constant temperature water bath for 5.5 h, adjust the pH to neutral, and the etherification reaction ends. Subsequently, place it in an activated dialysis bag with a cut-off molecular weight of 8 - 14 KD, use ultrapure water as the permeation medium for purification, and change the exudate every 3 h; after purification, filter by suction and rotary evaporation, and finally freeze-dry for 48 h to obtain CBSP.

[0068] Example 4 Screening of pH in the Preparation Process of CBSP-CP

[0069] Respectively taking the viscosity retention rate, yield value, and transparency as indicators, investigate the pH conditions for preparing the composition.

[0070] Determination of viscosity retention rate Adjust the initial viscosities of the CBSP-CP gel matrix composition gels with various ratios to similar values. Weigh 5 g of different gel matrix composition gels, add deionized water and NaCl solution respectively, ensuring that the mass fractions of NaCl are 0%, 0.3%, respectively. After stirring until NaCl is completely dissolved in the gel sample, use a DV-III rheometer, mode 2, at room temperature of 25 °C, 64# rotor, and a rotation speed of 10 rpm to measure the gel viscosity, denoted as ηwater and ηsalt. Each sample is measured in parallel three times and the average value is taken. Calculate the viscosity retention rate μ. The larger the μ value, the better the ion resistance of the gel matrix.

[0071] Determination of yield value Take the CBSP-CP gel sample added with 0.3% mass fraction of NaCl, use a DV-III rheometer, mode 2, 64# rotor, and measure the viscosities η0.5 and η1 at rotation speeds of 0.5 rpm and 1 rpm respectively, and calculate the yield stress, that is, the yield value (γ0).

[0072] Determination of transparency For the prepared CBSP-CP sample, use a UV-6100 type ultraviolet spectrophotometer to measure the transmittance at 425 nm with ultrapure water as the reference. The size of the transmittance can characterize the transparency of the gel.

[0073] The applicant's experiment shows that CBSP has a stable positive charge amount in the range of pH 5 - 9. Therefore, prepare CBSP-CP gel matrix compositions with pH values of 5, 6, 7, 8, and 9. To avoid the influence of metal ion factors, use a citric acid saturated solution and triethanolamine to adjust the pH. The results are shown in Table 1.

[0074] Table 1 Influence of pH on the Ion Resistance of CBSP-CP Gel Matrix Composition

[0075]

[0076] As can be seen from the results in Table 1, under different pH conditions, the viscosity retention rate and yield value of the formed CBSP-CP matrix composition are different. The CBSP-CP matrix composition formed under the condition of pH 7-9 has better viscosity retention rate and yield value; while the transparency shows a trend of first increasing and then decreasing, and the transparency is better in the range of pH 5-8. When pH is 6-7, the formed CBSP-CP composition has the highest transparency.

[0077] Example 5 Screening of the Preparation Temperature of CBSP-CP

[0078] Taking 25, 35, 50, 60, 70 °C as the compounding temperature, the effect of the preparation temperature on the preparation of the CBSP-CP matrix composition was investigated. The results are shown in Table 2.

[0079] Table 2 Effect of Temperature on the Preparation of CBSP-CP Gel Matrix Composition

[0080]

[0081] As can be seen from Table 2, the CBSP-CP matrix composition prepared at 25-60 °C has better viscosity retention rate, yield value and transparency, and shows a gradually decreasing trend. It may be that high temperature has a certain influence on the formation of the CBSP-CP composite structure. Therefore, 25-35 °C is the best.

[0082] Example 6 Screening of the Concentration of the CBSP-CP Preparation Process

[0083] Under the conditions of pH 7 and 25 °C, taking the viscosity retention rate, yield value, transparency and centrifugal delamination as indexes, the effects of different combined mass concentrations on the preparation of the CBSP-CP composition were investigated respectively. The results are shown in Table 3.

[0084] Table 3 Effect of Concentration on the Preparation of CBSP-CP Matrix Composition

[0085]

[0086] As can be seen from Table 3, the CBSP-CP composition prepared with a combined mass concentration of 1-1.5% has good performance; when the concentration is too high (2-3%), the formed CBSP-CP composition has high viscosity and excessive yield value. The appropriate concentration can be selected within the range of 1-1.5% according to needs.

[0087] Example 7 CBSP-CP Composite Ratio

[0088] According to the above experimental results, under the conditions of the optimal concentration, pH and temperature, CBSP∶CP was combined at ratios of 1∶3, 1∶2, 1∶1, 2∶1, 3∶1 (w / w), and the results are shown in Table 4. When CBSP∶CP was less than 1∶2, the viscosity retention rate, yield value and transparency of the prepared CBSP-CP composition decreased significantly after adding ions, and it was prone to delamination after centrifugation; when the ratio of CBSP∶CP was 1∶1 - 3∶1, the μ, γ0 and T values of the prepared CBSP-CP increased significantly with the increase of the CBSP ratio, and the optimal ratio was 1∶1 - 2∶1 for CBSP∶CP. The results are shown in Table 4.

[0089] Table 4 Influence of combined ratio on the preparation of CBSP-CP composition

[0090]

[0091]

[0092] Example 8 Preparation of combinations of CBSP and different types of CP

[0093] Under the conditions of the optimal concentration, pH and temperature, CBSP was combined with CP 940, CP 2020, and CP U20 at a ratio of 2∶1 (w / w) respectively, and the preparation of combinations of CBSP and different types of CP was investigated. The results are shown in Table 5.

[0094] The results showed that CBSP-CP compositions could be prepared by combining CBSP with anionic CP under certain conditions. Among them, the novel hydrophobically modified Carbopol ETD 2020 could endow the composition with more constant viscosity and stable ion resistance compared with the traditional polyacrylic acid-based Carbopol 940 and the long-chain modified C10-30 alkanol acrylate cross-linked polymer Carbopol Ultrez 20. In addition, the wetting and dispersion of the CBSP-CP2020 composition were faster than those of CBSP-CP 940 and CBSP-CP U20.

[0095] Table 5 Influence of CP type on the preparation of CBSP-CP composition

[0096]

[0097] Example 9 Preparation of CBSP-CP (1∶1)

[0098] Disperse 0.5% (w / v) CBSP in deionized water at room temperature of 25°C, and stir at 400 rpm until uniform to form a CBSP phase;

[0099] Disperse 0.5% (w / v) CP 2020 in deionized water at 70 °C, stir at 400 rpm until uniform, and let stand for 12 h until a uniform and lump-free sol is formed to obtain the CP phase;

[0100] Slowly add the CBSP phase to the CP phase, and adjust the pH using a saturated solution of citric acid and triethanolamine to obtain the product.

[0101] Preparation of CBSP-CP (2:1) with a concentration of 1% (w / v) in Example 10

[0102] Disperse 0.33% (w / v) CP 2020 in deionized water at 70 °C, stir at 400 rpm until uniform, and let stand for 12 h until a uniform and lump-free sol is formed, which is called Phase A;

[0103] Disperse 0.66% (w / v) CBSP in deionized water at room temperature (25 °C), stir at 400 rpm until uniform to form a CBSP solution, which is called Phase B;

[0104] At room temperature (25 °C), slowly add Phase B to Phase A, adjust the pH to 7 using a saturated solution of citric acid and triethanolamine, and stir until uniform to obtain the product.

[0105] Preparation of CBSP-CP (2:1) in Example 11

[0106] Disperse 0.5% (w / v) CP 2020 in deionized water at 70 °C, stir at 400 rpm until uniform, and let stand for 12 h until a uniform and lump-free sol is formed, which is called Phase A;

[0107] Disperse 1.0% (w / v) CBSP in deionized water at room temperature (25 °C), stir at 400 rpm until uniform to form a CBSP solution, which is called Phase B;

[0108] At room temperature (25 °C), slowly add Phase B to Phase A, adjust the pH to 6.5 using a saturated solution of citric acid and triethanolamine, and freeze-dry to obtain the CBSP-CP matrix composition.

[0109] Preparation of CBSP-CP (3:1) in Example 12

[0110] Disperse 0.25% (w / v) CP 2020 in deionized water at 80 °C, stir at 500 rpm until uniform, and let stand for 12 h until a uniform and lump-free sol is formed, which is called Phase A;

[0111] Disperse 0.75% (w / v) CBSP in deionized water at room temperature (25 °C), stir at 400 rpm until homogeneous to form a CBSP solution, designated as Phase B; at room temperature, slowly add Phase B to Phase A, and adjust the pH to 8 using a saturated citric acid solution and triethanolamine; dry under vacuum to obtain a white to off-white CBSP-CP matrix composition.

[0112] Preparation of CBSP-CP (3:1) in Example 13

[0113] Disperse 0.35% (w / v) CP 2020 in deionized water at 80 °C, stir at 500 rpm until homogeneous, and let stand for 12 h until a homogeneous, lump-free sol is formed, designated as Phase A; adjust the pH to 8 using a saturated citric acid solution and triethanolamine.

[0114] Disperse 1.05% (w / v) CBSP in deionized water at room temperature (25 °C), stir at 400 rpm until homogeneous to form a CBSP solution, designated as Phase B;

[0115] At room temperature, slowly add Phase B to Phase A and stir until homogeneous to obtain.

[0116] Preparation of CBSP-CP (1:3) in Example 14

[0117] Disperse 0.9% (w / v) CP 2020 in deionized water at 60 °C, stir at 300 rpm until homogeneous, and let stand for 12 h until a homogeneous, lump-free sol is formed, designated as Phase A;

[0118] Disperse 0.3% (w / v) CBSP in deionized water at room temperature (25 °C), stir at 400 rpm until homogeneous to form a CBSP solution, designated as Phase B;

[0119] At 25 °C, slowly add Phase B to Phase A; adjust the pH to 6 using a saturated citric acid solution and triethanolamine to obtain.

[0120] Preparation of CBSP-CP (1:2) in Example 15

[0121] Disperse 0.8% (w / v) CP 2020 in deionized water at 60 °C, stir at 300 rpm until homogeneous, let stand for 12 h until a homogeneous, lump-free sol is formed, and adjust the pH to 6 using a saturated citric acid solution and triethanolamine, designated as Phase A;

[0122] Disperse 0.4% (w / v) CBSP in deionized water at room temperature (25 °C), stir at 400 rpm until homogeneous to form a CBSP solution, designated as Phase B;

[0123] At 25 °C, slowly add Phase B to Phase A to obtain a CBSP-CP matrix composition.

[0124] Preparation of Example 16 CBSP-CP (2:1)

[0125] Disperse 0.4% (w / v) Carbopol 940 in deionized water at 60 °C, stir at 300 rpm until homogeneous, let stand for 12 h until a homogeneous, lump-free sol is formed, and adjust the pH to 6 using a saturated solution of citric acid and triethanolamine, designated as Phase A;

[0126] Disperse 0.8% (w / v) CBSP in deionized water at room temperature (25 °C), stir at 400 rpm until homogeneous to form a CBSP solution, designated as Phase B;

[0127] At 30 °C, slowly add Phase B to Phase A to obtain a CBSP-CP matrix composition.

[0128] Preparation of Example 17 CBSP-CP (1:1)

[0129] Disperse 0.8% (w / v) Carbopol 1342 in deionized water at 60 °C, stir at 300 rpm until homogeneous, let stand for 12 h until a homogeneous, lump-free sol is formed, and adjust the pH to 6.3 using a saturated solution of citric acid and triethanolamine, designated as Phase A;

[0130] Disperse 0.8% (w / v) CBSP in deionized water, stir at 400 rpm until homogeneous to form a CBSP solution, designated as Phase B;

[0131] At 35 °C, slowly add Phase B to Phase A, stir well, and freeze-dry to obtain a CBSP-CP matrix composition.

[0132] Preparation of Example 18 CBSP-CP (1:2)

[0133] Disperse 0.8% (w / v) Carbopol 1342 in deionized water at 60 °C, stir at 300 rpm until homogeneous, let stand for 12 h until a homogeneous, lump-free sol is formed, and adjust the pH to 6.5 using a saturated solution of citric acid and triethanolamine, designated as Phase A;

[0134] Disperse 0.4% (w / v) CBSP in deionized water at room temperature (25 °C), stir at 400 rpm until homogeneous to form a CBSP solution, designated as Phase B;

[0135] At 30 °C, slowly add Phase B to Phase A, stir well, and freeze-dry to obtain a CBSP-CP matrix composition.

[0136] Preparation of Example 19 CBSP-CP (2:1)

[0137] Disperse 0.4% (w / v) CPU20 (Carbopol Ultrez 20) in deionized water at 60 °C, stir at 300 rpm until homogeneous, let stand until a homogeneous, lump-free sol is formed, and adjust the pH to 6.5 using saturated citric acid solution and triethanolamine, which is designated as Phase A;

[0138] Disperse 0.8% (w / v) CBSP in deionized water, stir at 400 rpm until homogeneous to form a CBSP solution, which is designated as Phase B;

[0139] At 35 °C, slowly add Phase B to Phase A, stir well, and dry under vacuum to obtain the CBSP-CP matrix composition.

[0140] Example 20 Infrared Spectral Analysis of Bletilla striata Polysaccharide (BsP) and Cationized Bletilla striata Polysaccharide (CBSP)

[0141] Use a TENSOR 27 Fourier transform infrared spectrometer to detect the infrared spectra of Bletilla striata polysaccharide and the modified cationized Bletilla striata polysaccharide (Example 3). In the experiment, the potassium bromide (KBr) powder pressing method was used for testing. Weigh the test substance and KBr in a ratio of 1:100 in an agate mortar, grind and mix well, then take an appropriate amount of the mixed powder into a tablet pressing mold, and press it into a transparent thin film under a pressure of 10 MPa. Subsequently, scan the images of BSP before and after the reaction in the wavenumber range of 500 - 4000 cm -1 The results are shown in Figure 1 .

[0142] From Figure 1 it can be seen that: compared with BSP, new characteristic peaks appear in the modified CBSP. For example, the stretching vibration peak of the C-H bond on the quaternary ammonium salt of CBSP is at 1670 cm -1 ; a new peak appears at 1568 cm -1 , corresponding to the absorption peak of the C-N bond on the quaternary ammonium salt.

[0143] Example 21 1 1H-NMR

[0144] Experimental conditions: Weigh approximately 10.00 mg of cationized Bletilla striata polysaccharide (CBSP) (Example 3) and dissolve it in 600 μL of D2O. After vortexing, transfer it to a nuclear magnetic resonance tube respectively, and measure the 1H-NMR spectrum using an AVANCE III nuclear magnetic resonance spectrometer under the conditions of a temperature of 293 K and a frequency of 500 MHz.

[0145] The 1 1H-NMR spectrum of CBSP is shown in Figure 2As shown in the figure. It can be analyzed by Mestrenova software that the peak at the chemical shift δ = 4.80 ppm belongs to the solvent heavy water; from the enlarged view, the chemical shifts of some hydrogen atoms of CBSP are δ 3.22 ppm (H1), 3.48 ppm (H3), and 3.58 ppm (H2) respectively. Among them, the peak at δ = 3.22 ppm belongs to the -CH3 group of the quaternary ammonium salt group, indicating that the etherification reaction occurs between BSP and CTA in the homogeneous system, and the cationic group is attached to the BSP molecular chain.

[0146] Example 22 Thermogravimetric (TGA) Analysis of CBSP and BSP

[0147] TGA experimental conditions: Weigh about 10.00 mg of fully dried BSP and CBSP respectively and place them in the aluminum crucibles for TGA analysis, then place them on the automatic sampling tray of the TGA. The samples are successively tested under the conditions of a nitrogen flow rate of 20 mL / min, a heating rate of 10 °C / min, and a heating range of 50 °C to 600 °C.

[0148] The thermogravimetric analysis results of BSP and modified CBSP are as Figure 3 (a, b) shown. The two curves of the thermogravimetric analysis of each sample respectively represent the changes of mass loss (TG) and derivative increment (DTG) with temperature, indicating the degradation trend and degradation rate of the sample. As can be seen from the figure, the pyrolysis process of BSP is as shown in 3(a). The first-stage thermal degradation occurs at 51.5 - 125 °C. After that, as the temperature rises, the mass of the sample remains constant until the temperature reaches 212 °C, and the TG of the system shows an inflection point, symbolizing the start of the second weight loss stage. In this stage, the mass of the sample decreases rapidly. When the temperature rises to 347 °C, the sample loses 52.63% of its mass. From the DTG curve, it can be obtained that T max , the rate of mass loss of the sample is the fastest at 250 °C. After the end of the second stage of sample weight loss, as the temperature rises, the mass of the sample does not decrease very significantly. Finally, at 600 °C, 15.86% of the residue remains. The thermal decomposition of the modified CBSP is as shown in Figure 3 (b). The T max of CBSP is 265 °C, which is 15 °C higher than that of BSP. And in the second-stage pyrolysis process, the mass of the original BSP powder decreased by 52.63%, while the cationized CBSP lost 46.10% and 31% of the residue remained. The results show that the thermal stability of BSP after cationic modification has been improved.

[0149] Example 23 Zeta Potential of CBSP at Different pH Values

[0150] Experimental conditions: The zeta potential of the surfaces of BSP and CBSP gels was measured using a Malvern particle size analyzer. BSP and CBSP were prepared into gels with a mass fraction of 2%. To avoid the influence of metal ion factors, a citric acid saturated solution and triethanolamine were used to adjust the pH. Before testing, 1 mL of the sample was diluted 20 times with deionized water, and a pipette was used to add it dropwise to the sample cell for measurement, to characterize the charge-carrying situation of cationic Bletilla striata polysaccharide under different pH conditions.

[0151] The results are as Figure 4 shown. Bletilla striata polysaccharide is almost uncharged when the pH is less than 7, and when the pH is greater than 7, the system carries a certain amount of negative charge; while cationic Bletilla striata polysaccharide carries a certain amount of positive charge under different pH conditions, which provides a guarantee for binding with negatively charged polyelectrolytes through electrostatic interactions, and within the pH range of 5 - 9, its potential value changes slightly, which is more conducive to the stability of the system.

[0152] Example 24 Comparison of the appearance properties of CBSP and BSP

[0153] An appropriate amount of CBSP, BSP, and CP2020 were fully swollen, and the formability, uniformity, and transparency of the resulting matrix gels were observed; CBSP and BSP were prepared into sols with different mass fractions, and a UV-6100 type ultraviolet spectrophotometer was used to measure the transmittance at 610 nm with ultrapure water as the reference. The greater the transmittance, the higher the transparency. It was measured three times in parallel and the average value was taken. The results are shown in Table 5. Figure 5 、 Figure 6 . The results show that the transparency of CBSP is basically the same as that of CP2020. Compared with BSP, the water solubility of CBSP increases, and the transparency of the solution is significantly improved.

[0154] Table 5 Appearance properties of CBSP and BSP

[0155]

[0156] Example 25 Scanning electron microscope results of the CBSP-CP composition

[0157] An appropriate amount of freeze-dried CP, CBSP, and the CBSP-CP of the present invention (Example 11) were taken, fully dried and fixed on a sample stage attached with conductive glue, a metal thin film was deposited using a vacuum ion sputtering instrument, and the microscopic morphology of the samples was observed using a scanning electron microscope under the condition of an acceleration voltage of 20 kV. The results are shown in Figure 7 .

[0158] Among them, A(a), B(b), and C(c) represent the SEM images of CBSP, CP, and CBSP-CP at 120X (600X), respectively. CBSP presents a loose and porous layered structure; CP is a sheet-like structure with uneven void distribution. When the magnification is 600X, it can be seen that the connection between the voids is not tight; and the CBSP-CP composition of the present invention presents a dense, relatively evenly distributed porous three-dimensional network structure, showing a different microstructure.

[0159] Example 26 Matrix properties of the CBSP-CP composition of the present invention - ion resistance

[0160] Comparison of ion resistance with the matrix of Bletilla striata polysaccharide plus carbomer (BSP+CP)

[0161] Take about 0.3g BSP and completely dissolve it in 90mL deionized water to prepare a BSP solution. Take about 0.15g CP 2020 and add it to the solution and heat it to 80°C for 5h. Use triethanolamine to adjust the pH value to 7 to prepare a BSP+CP 2020 matrix. According to the method of Example 10, take about 0.3g CBSP and 0.15g CP 2020 respectively to prepare a CBSP-CP 2020 combined matrix. The matrix is fully swelled, and 12 portions of each, 5g each, are taken, and deionized water, NaCl, and CaCl2 solutions are added respectively, so that the mass fraction of NaCl is 0, 0.1%, 0.3%, 0.5%, 0.7%, and 1%; the mass fraction of CaCl is 0, 0.1%, 0.3%, 0.5%, 0.7%, and 1%. The viscosity (η) of the matrix is measured, and the average value is taken for three times. The viscosity of the two matrices in NaCl is calculated. + , Ca 2+ Viscosity retention rate under ionic conditions (μ), where μ = η salt / η water × 100%. The larger the μ, the better the ionic resistance of the matrix. The results are shown in Tables 6 and 7.

[0162] The results showed that BSP+CP could only tolerate trace amounts of Na + , the viscosity decreases sharply at a slightly higher concentration, and it has almost no tolerance to divalent metals. However, CBSP-CP shows a stable viscosity retention rate in a wide concentration range, showing excellent ion resistance to monovalent ions and even divalent ions.

[0163] Table 6 Ion resistance of BSP+CP and CBSP-CP (Na + )

[0164]

[0165] Table 7 Ion resistance of BSP+CP and CBSP-CP (Ca 2+ )

[0166]

[0167] Comparison of the ion resistance performance with commercially available matrix materials

[0168] According to the method under 1.1, taking three matrix materials, namely traditional CP 940, improved (ion-resistant improved) CP 2020, and polyacrylate cross-linked polymer - 6 (zen), etc., as references, the ion resistance properties of the matrix of the CBSP-CP composition of the present invention were evaluated. Respectively take several matrix materials of CBSP-CP (hereinafter taking Example 11 as an example), CP940, CP2020, Zen, and CBSP. Referring to the rotational viscometer determination method in the Chinese Pharmacopoeia 2020 edition, through a DV-III rheometer, mode 2, at room temperature of 25 °C, 64 # rotor, under the condition of a rotational speed of 10 rpm, measure the viscosity value (η1o) of the sample three times and take the average value. The viscosity of the matrix material under different mass fractions of Na + , Ca2+ conditions are shown in Figure 8 (a), Figure 8 (b).

[0169] The results show that with the increase of ions, the viscosities of traditional CP 940 and improved CP 2020 both decrease sharply. When the mass fraction of NaCl is 0.5% and the mass fraction of CaCl2 is 0.3%, the viscosity even drops to 0, and the matrix completely loses its viscosity; the viscosity reduction of Zen is slightly slower than that of CP 940 and CP 2020, but when Ca 2+ is 0.3%, the viscosity also almost drops to 0. However, the viscosity of CBSP-CP almost remains unchanged with the increase of Naw, Ca 2+ concentration. Even under the conditions of high ion content of 1.5% NaCl and 1.0% CaCl2, the viscosity remains almost stable, and the viscosity retention rate is still greater than 90%, indicating that the matrix of the CBSP-CP composition has excellent ion resistance performance.

[0170] Example 27 Matrix performance of the CBSP-CP composition of the present invention - Rheological properties

[0171] Taking three matrix materials, namely CP 940, CP 2020, and Zen, as references, the rheological properties of the matrix of the CBSP-CP composition of the present invention were evaluated.

[0172] The rheological curve was measured using a DV-III rheometer, mode 1, SC4-34# rotor, at room temperature of 25 °C, and within the range of shear rate 0.01 - 70S -1 to measure the viscosity and shear stress of the matrix and draw the change curve of viscosity - shear stress. The change curves of shear stress and viscosity of different matrix materials with shear rate are shown inFigure 9 As shown, the shear stress-shear rate curve of CBSP-CP is similar to that of CP 940 and Zen, and the viscosity-shear rate curve is similar to that of CP 940, CP 2020, and Zen. The Cross equation was used to fit the shear rate-viscosity curve, and the rheological parameters of each matrix are shown in Table 8. The results show that the flow indices n of CP 940, CP 2020, Zen, and CBSP-CP of the present invention are all between 0 and 1, and they all belong to non-Newtonian pseudoplastic fluids; from the analysis of the time characteristic parameter t, it can be seen that the rate of CBSP-CP from zero-shear viscosity to infinite-shear viscosity is similar to that of CP 940 and CP 2020, with strong fluidity and easy spreading; while the time parameter of Zen from zero-shear viscosity to infinite-shear viscosity is larger, and its fluidity is relatively weak and its ductility is poor. Cross equation: In the formula, η0 is the zero-shear viscosity, η ∞ is the infinite-shear viscosity, t is the time characteristic parameter, n is the flow index,

[0173] Table 8 Fitting parameters of shear rate-viscosity models for different matrices

[0174]

[0175] Thixotropy Thixotropy refers to the property that the viscosity of the gel matrix decreases under shear action and can increase and recover after the shear stops. Good thixotropy is beneficial to endowing the gel with the ability to be easily dispersed on the coated surface such as the skin surface during the application process under a certain shear stress, and is also beneficial to its retention on the coated surface. A good matrix should have a certain thixotropy to ensure that the gel has appropriate spreading performance during application and can maintain its shape when standing still. Using a DV-III rheometer, mode 1, SC4-34# rotor, at room temperature of 25°C, i set the shear rate to linearly increase from 0.1 s -1 to 70 s within 100 s -1 and then decrease from 70 s to 0.1 s w at the same rate within 100 s -1 to measure the change of shear stress with shear rate. The measurement results of different matrices are shown in l Figure 10 .

[0176] Figure 10 The downward line and the upward line of the matrix do not completely coincide, forming a hysteresis loop with different areas, showing a certain positive thixotropy. From Figure 10From the area of the mid-hysteresis loop, it can be seen that CP 940, CP 2020 and the CBSP-CP of the present invention all have a certain positive thixotropy. Their internal structures have a reduced viscosity under the action of shear force and can immediately return to their original state when the shear stress decreases or disappears. That is, during the use of the gel, the viscosity is reduced by pumping and spreading, which is beneficial for spreading, and the original structure can be restored in time when the use stops; while the thixotropy of Zen is relatively weak.

[0177] Safety - Biocompatibility of the CBSP-CP Composition in Example 28

[0178] Biocompatibility is a method to characterize the interaction between materials and the body. In this study, the most commonly used blood compatibility test was adopted. ① Preparation of blood cell dispersion: Fresh rabbit blood was collected with a disposable vacuum blood collection tube, gently shaken to mix it with the anticoagulant in the tube, and then mixed with physiological saline. It was centrifuged at 1500 rpm for 10 min, and the supernatant was discarded; then it was rinsed with physiological saline and centrifuged until the supernatant became transparent, and the supernatant was discarded. The red blood cells were collected and prepared into a 2% blood cell suspension with physiological saline, which was called Solution A. ② Preparation of test samples: 0.2 g of Zen, CBSP-CP, CP 2020 and CP 940 matrix samples were weighed and dissolved in 10 mL of physiological saline respectively, and diluted successively to prepare gel dispersions with concentrations of 20, 10, 5, and 1 mg / mL respectively, which were called Solution B. ③ Preparation of experimental group, positive control and negative control group samples: Solution A and Solution B were mixed at a ratio of 1:1 as the experimental group, and incubated at 37 °C for 2 h. After centrifugation (2000 rpm, 5 min), 200 μL was taken and added to a 96-well plate, and the OD value of the supernatant was measured at a wavelength of 545 nm using an enzyme-labeled instrument. Solution A was mixed with equal amounts of deionized water and physiological saline respectively as the positive control and negative control. Each experiment was repeated three times in parallel, and the mean value was taken. The hemolysis rate (%) was calculated using the following formula:

[0179]

[0180] Generally, the lower the hemolysis rate, the smaller the degree of red blood cell fragmentation and the lower the possibility of direct hemolysis. According to the national evaluation standard in China, a hemolysis rate lower than 5% is considered to meet the requirements.

[0181] The results are as Figure 11 shown. After standing at 37 °C for 2 h and centrifugation, the supernatants prepared from the four matrices at low concentrations (1, 5, 10 mg / mL) were all colorless and clear, and the supernatants of Zen and CP 2020 at high concentration (20 mg / mL) were slightly red. The red blood cells of the samples at different concentrations all sank to the bottom of the tube, similar to the negative control group; while the samples in the positive control group were bright red and there was no red blood cell residue at the bottom.

[0182] The results of the hemolysis test of different matrices are shown in Figure 12, at low concentrations, the hemolysis rates of all four matrices are lower than the specified value of 5%. Among them, the hemolysis rates of CBSP-CP and CP 940 at high concentrations are also less than 5%, indicating that CBSP-CP has good biocompatibility and the composition is superior to CP 2020.

[0183] Example 29 Safety of CBSP-CP Composition - Skin Safety

[0184] Referring to the commonly used acute skin irritation test in new drug research and development, using three matrix materials, namely CP 940, CP 2020, and Zen, as references, the skin safety of the CBSP-CP composition matrix was evaluated. Fifteen healthy SD rats were randomly divided into 5 groups. 24 hours before the experiment, the fur on both sides of the rat spine was removed, and the depilated areas on the left and right were each 2×5 cm 2 , before administration, the administration site was washed with warm water to ensure that the depilated skin was not damaged. Using the self-contrast method of the left and right sides of the same body, 0.5 g of the matrix sample was evenly applied to the left side, and ultrapure water was applied to the right side as a blank control. At 1 h, 24 h, 48 h, and 72 h after administration, the rats' skin was observed for edema, erythema, skin roughness, bleeding, etc.

[0185] Experimental conditions: temperature (22±1°C), relative humidity (60 - 70%), and the test animals should be adapted to this environment for at least 3 days before the experiment.

[0186] Skin irritation reaction evaluation criteria. Referring to the skin adverse reaction scoring criteria (see Table 9), the test score results were recorded, and the irritation intensity score was calculated using the following formula, and then the irritation intensity of the sample to the skin was judged according to Table 10.

[0187]

[0188] Table 9 Skin irritation reaction scoring criteria

[0189]

[0190] Table 10 Skin irritation intensity scoring criteria

[0191]

[0192] After single application of the four matrices and purified water, the test area was observed for any abnormalities, and scored according to Tables 9 and 10. The results are shown in Figure 13 , Table 11.

[0193] Table 11 Investigation results of skin irritation of different matrix materials (n = 3)

[0194]

[0195]

[0196] The stimulation intensity scores of CBSP-CP and other commercially available matrix materials are all 0, and no phenomena such as erythema and edema are observed, and there is no acute stimulation reaction.

[0197] Application of the matrix material of the CBSP-CP composition in Example 30 in a gelling agent containing metal ions and complex components

[0198] Using dandelion-scutellaria baicalensis extract and zinc gluconate as models containing metal ions and complex traditional Chinese medicine components, using commercially available CP 940, CP 2020, Zen and the CP-CBSP of the present invention (Example 17) as matrix materials respectively, with the same other components and preparation methods, four kinds of Pu-Huang-Zinc gels under different matrix conditions were prepared to investigate the influence of the matrix material on the performance and quality of the gelling agent containing metal ions and complex components.

[0199] Prescription composition:

[0200]

[0201]

[0202] Take an appropriate amount of glycerol as a wetting agent, and fully swell with four matrixes of CP 940, CP 2020, Zen and CP-CBSP (Example 17) and the moisturizer sodium hyaluronate, add an antioxidant and dandelion-scutellaria baicalensis extract (A), and stir until uniform; prepare a zinc gluconate solution (B), mix (B) into (A), then add a penetration enhancer, surfactant, preservative, etc. and stir well, supplement deionized water to a fixed volume, adjust the pH, and add an appropriate amount of essence to prepare Pu-Huang-Zinc gels with different matrixes.

[0203] Example 31 Influence of CP-CBSP and commercially available matrixes on the stability of the compound traditional Chinese medicine Pu-Huang-Zinc gel

[0204] The samples in Example 30 were all encapsulated in colorless transparent vials and placed under different temperatures and strong light (4500 lx) conditions respectively to investigate the changes in the appearance properties, viscosity and the content of the index components of dandelion (chicoric acid) and scutellaria baicalensis (baicalin) of the Pu-Huang-Zinc gels prepared with different matrix materials. The results are shown in Tables 12-15. In the tables, "↓" and "↑" represent the degree of decrease or increase compared with day 0. The results show that the compound Pu-Huang-Zinc gel prepared with the CBSP-CP of the present invention as the matrix material has better stability in terms of the appearance properties, viscosity and the content of the active index components under high temperature, low temperature and strong light conditions (P < 0.01).

[0205] Table 12 Stability of Pu-Huang-Zinc gels of CP-CBSP and commercially available matrixes (high temperature 40 °C) (n = 3)

[0206]

[0207]

[0208] Table 13 Stability of CP-CBSP and commercially available matrix Pollen Typhae-Zinc Gel (at 60°C high temperature) (n = 3)

[0209]

[0210] Table 14 Stability of CP-CBSP and commercially available matrix Pollen Typhae-Zinc Gel (at -4°C low temperature) (n = 3)

[0211]

[0212]

[0213] Table 15 Stability of CP-CBSP and commercially available matrix Pollen Typhae-Zinc Gel (under strong light of 4500 lx) (n = 3)

[0214]

[0215] Example 32 Influence of CP-CBSP and commercially available matrix on the rheological properties of gel - Steady-state viscosity

[0216] Using a DV-III rheometer, mode 1, SC4-34# rotor, shear at a constant rate of 20 s -1 for 120 s at room temperature of 25°C to measure the steady-state apparent viscosity of the gel sample. The results of the steady-state viscosity of the compound Pollen Typhae-Zinc Gel prepared with different matrix materials are shown in Table 16. Compared with the blank matrix, the compound Pollen Typhae-Zinc Gel containing complex components of metal ions and traditional Chinese medicine extracts prepared with CBSP-CP of the present invention has the smallest decrease in steady-state apparent viscosity, indicating that CBSP-CP has a better viscosity-increasing effect and can maintain the viscosity of the system for the complex-component gel.

[0217] Table 16 Results of steady-state viscosity of CP-CBSP and commercially available matrix compound Pollen Typhae-Zinc Gel

[0218]

[0219] Example 33 Influence of CP-CBSP and commercially available matrix on the rheological properties of gel - Rheological curve

[0220] The rheological curves of the compound Pollen Typhae-Zinc Gel prepared with different matrix materials are shown in Figure 14 . At room temperature (25°C) and body surface temperature (37°C), each gel shows that the viscosity decreases with the increase of shear rate, conforming to the shear-thinning pseudoplastic fluid, which is beneficial for spreading. In the range of 0.01 - 70 S -lWithin the shear range, the viscosity value of the compound gel based on CBSP-CP of the present invention always remains at a relatively high level, and the terminal viscosity value is still greater than 1000 mPa·s, indicating that CBSP-CP has better shear resistance when applied to ionic and complex component systems.

[0221] Effect of CP-CBSP and commercially available matrices on percutaneous penetration characteristics of compound Puhuang-Xin gel

[0222] Referring to the literature method, using excised rat skin as a barrier model, the prepared rat skin was washed with normal saline before the experiment, dried with absorbent paper and then fixed in a diffusion cell; using normal saline containing 20% ethanol as the receiving medium, rotation speed: 400 r / min, temperature: 37 ± 0.1 °C; using baicalin as the index component, 1 g of compound Puhuang-Xin gel samples containing CP-CBSP and commercially available matrices were added to the supply cell respectively, with 4 parallels in each group. Samples of 2 mL were taken at different times, and an equal volume of blank receiving solution at the same temperature was replenished. The content of baicalin was determined by HPLC method, and the cumulative penetration amount (Q, μg·cm -2 ) and cumulative permeability (F, %) were calculated by the following formulas 1 and 2. After the 24-hour percutaneous penetration experiment, the rat skin was removed, washed with normal saline, the residual moisture was removed by suction, cut into pieces, 5 mL of methanol was added, homogenized for 5 min, sonicated for 30 min, centrifuged, and the supernatant was taken to determine the content of baicalin. The skin retention rate (Qs) was calculated according to formula 3.

[0223]

[0224]

[0225] In the formula, s is the effective diffusion area (3.14 cm 2 ); P n is the sampling concentration at the nth time point; V is the total volume of the receiving cell (8.1 mL); P i is the sampling concentration at the ith time point; C is the mass concentration of baicalin in the Puhuang-Xin gel; V0 is the sampling volume (2 mL); V1 is the total volume of the administered gel extract (10 mL).

[0226]

[0227] In the formula, C1 is the mass concentration of baicalin in the skin extract, V1 is the volume of the skin extract (5 mL), C is the mass concentration of baicalin in the compound Puhuang gel, and V is the total volume of the administered gel solution (10 mL).

[0228] The results of the change curves of the cumulative permeability (F) of baicalin in compound Puhuang-Xin gels containing CP-CBSP and commercially available matrices with time (t) are as Figure 15As shown in the figure, the 12-hour cumulative permeation rates of different matrix compound gel samples are different. Among them, the highest value of CBSP-CP reaches 73.31±2.84%, and the lowest value of CP 940 is 49.12±2.87%. The skin retention results are shown in Figure 16 , and the 12-hour skin retention amount of baicalin in the compound gel sample with CBSP-CP as the matrix is relatively large, which is conducive to forming a drug reservoir on the skin and exerting a long-term effect.

[0229] Example 35 Effects of CP-CBSP and commercially available matrices on the moisturizing effect of Puhuang Zinc Gel

[0230] Select healthy volunteers without a history of skin diseases. After sitting quietly in a constant temperature and humidity room (temperature: 22±0.5°C, relative humidity: 50±2%) for 30 minutes to adapt to the environment, measure the skin moisture content (MMV) and transepidermal water loss (TEWL) as the baseline values. Before the test, clean the inner sides of the left and right forearms with warm water and mark 5 square areas (4 cm×4 cm). The blank control group and the test sample group are randomly distributed in the test areas. Take the compound Puhuang Zinc Gel prepared with different matrices and apply it to different test areas with the index finger at a sample amount of 2 mg / cm 2 . Apply the sample at a coating speed of 2 circles per second. Use a Comermeter CM825 type skin moisture meter to measure the change in skin water content within 5 hours after applying the gel sample, and analyze the hydration state of the stratum corneum. Take the average value of 5 measurements in the test area as the test result; use a Tewameter TM Hex type skin water loss meter to measure the change in transepidermal water loss within 5 hours after applying the gel sample. Each time, measure at the same position and take the average value measured within 30 seconds as the test result. The skin water content-time curve of the compound Puhuang Zinc Gel with different matrices is shown in Figure 17 , and the skin water loss-time curve is shown in Figure 18 . MMV can reflect the change in the water content of the stratum corneum during the test period. The larger the value, the more the sample can enhance the skin hydration effect and improve the water replenishment effect; TEWL reflects the amount of water diffusing outward through the stratum corneum during the test period and can characterize the effect of the gel sample on the water-locking performance of the stratum corneum. The smaller the TEWL, the less water is lost through the stratum corneum of the skin, indicating that the sample can improve the skin barrier function and increase the water-locking effect of the stratum corneum. The measurement results of skin water content and transepidermal water loss show that compared with the commercially available matrix, the skin water content (MMV) of the compound gel with CBSP-CP as the matrix is significantly increased at each time point (P<0.05); the transepidermal water loss (TEWL) is significantly decreased (P<0.01), indicating that the CBSP-CP composition of the present invention as a gel matrix helps to improve the skin barrier function and enhance the moisturizing effect.

Claims

1. A composition, characterized in that It includes cationized Bletilla striata polysaccharide and an anionic polymer, and the anionic polymer is carbomer.

2. The composition according to claim 1, characterized in that The carbomer includes Carbopol 940, Carbopol 1342, Carbopol ETD 2020, and Carbopol Ultrez 20.

3. The composition according to claim 2, characterized in that The carbomer is Carbopol ETD 2020.

4. The composition according to claim 1, wherein the mass ratio of the cationized Bletilla striata polysaccharide to the anionic polymer is (1-3):(1-3).

5. The composition according to claim 4, wherein the mass ratio of the cationized Bletilla striata polysaccharide to the anionic polymer is 2:

1.

6. A method for preparing the composition according to any one of claims 1-5: Disperse the carbomer in water, stir and dissolve to obtain phase A; Disperse the cationized Bletilla striata polysaccharide in water, stir and dissolve to obtain phase B; Slowly add phase B to phase A to obtain the composition; or dry and store for later use.

7. The composition according to any one of claims 1-5, wherein the cationized Bletilla striata polysaccharide is characterized in that: its infrared spectrum has a peak at 1670 cm -1 and a peak at 1568 cm -1 .

8. The preparation method of the cationized Bletilla striata polysaccharide in the composition according to any one of claims 1-5: Alkalization reaction: Among them, R1 is selected from H and Na; Etherification reaction: Among them, R2 is selected from H, .

9. The application of the composition according to any one of claims 1-5 in the preparation of pharmaceuticals and chemicals.

10. The application of the composition according to any one of claims 1-5 in the preparation of a gelling agent.

11. The application of the composition according to any one of claims 1-5 as a gelling agent matrix.

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