Aluminum hydroxide adjuvant with boehmite structure as well as preparation method and application of aluminum hydroxide adjuvant

Abstract

The invention provides an aluminum hydroxide adjuvant with a boehmite-like structure and a preparation method and application thereof.The preparation method comprises the steps that under the stirring condition, an aluminum salt solution and a sodium hydroxide solution are pumped into a 0.05%-0.5% sodium chloride solution heated to the set temperature at the same time, the pumping speed is adjusted, stirring is conducted for 1-2 h, and then the aluminum hydroxide adjuvant with the boehmite-like structure is obtained; controlling the ratio of hydroxyl to aluminum ions in the reaction system to be 2.5-3.5; when the concentration of aluminum ions in the reaction system reaches 0.07-0.09 M, stopping pumping the aluminum salt solution into the reaction system, continuously pumping the sodium hydroxide solution, and stopping pumping the sodium hydroxide solution when the pH value of the reaction system reaches 7.8-8.5; and adding 0.05-0.5% of a sodium chloride solution into the reaction system to a preparation amount, carrying out a hydrothermal reaction, discarding the supernatant, and carrying out moist heat sterilization to obtain the aluminum hydroxide adjuvant with the boehmite structure. The preparation method is reliable in process, good in batch-to-batch consistency and stable in quality.

Description

Technical Field

[0001] The invention belongs to the field of vaccine preparations, and in particular relates to an aluminum hydroxide adjuvant with a pseudo-boehmite structure and a preparation method and application thereof. Background Art

[0002] With the development of biotechnology and people's attention to disease prevention, people take active measures such as vaccination to prevent diseases caused by some pathogens. In recent years, vaccine research and development technology has developed rapidly, and new technologies have been used to develop new vaccines such as new inactivated vaccines, subunit vaccines with clearer antigenic components, and other new vaccines such as anti-idiotypic antibody vaccines, nucleic acid vaccines, and synthetic peptide vaccines. These vaccine antigens have high purity and small relative molecular mass, but their immunogenicity is generally weak, and adjuvants need to be added to enhance the immune response induced by them. Among them, aluminum salt adjuvant is the most widely approved adjuvant for human vaccines, with a history of nearly 100 years of use, and its safety and effectiveness have been fully verified worldwide.

[0003] Aluminum hydroxide adjuvant is a type of aluminum adjuvant, which is widely used in bacterial vaccines and viral vaccines. Aluminum hydroxide adjuvant is generally prepared by the alkaline method, such as adding alkaline substances (sodium hydroxide or ammonia water) to the aluminum salt solution to generate a neutral product without charge. As a vaccine adjuvant, aluminum hydroxide adjuvant generally requires the particle size to be controlled at around 10μm, with a larger specific surface area, in order to better exert the adjuvant effect and not cause local inflammatory reactions. Although aluminum adjuvants have been widely used in the vaccine field for decades, only Croda's Adjuvants and Canadian Chemtrade Adjuvants can meet the above physical and chemical properties. Due to technical confidentiality, there are almost no preparation patents or related literature for these two adjuvants. There is no finished aluminum adjuvant production and sales, and most vaccine manufacturers prepare, produce and use them by themselves. The preparation method of aluminum adjuvant has a great impact on its quality. Even if the same method is used to prepare aluminum adjuvant, there are certain differences in the production process of each manufacturer.

[0004] CN113559255 A discloses a method for obtaining an aluminum hydroxide adjuvant by adding an aluminum chloride solution to an ammonia solution in the form of a spray. CN108066759 A discloses a method for preparing an aluminum hydroxide adjuvant by dripping sodium hydroxide into a potassium aluminum sulfate solution. CN104367998 A discloses a prepared aluminum hydroxide adjuvant, wherein the particle size less than 10 μm accounts for 55.53%, and the adsorption capacity is 3.5 mg BSA / mg Al. Although some parameters can meet the requirements of aluminum adjuvants, the temperature changes greatly in its preparation process, the heat treatment time is as long as 5 hours, and then it is heat treated at a second temperature for 1-43 hours. The process is complicated and time-consuming, which is not conducive to the production of production enterprises. The particle size and distribution of the aluminum hydroxide adjuvants prepared by the above invention patents vary greatly, and the process is unstable, and there is no report on the size of the preparation amount.

[0005] In summary, there is an urgent need to find a preparation process for aluminum hydroxide adjuvant for vaccines that is reliable, has good batch consistency, stable quality and is easy to scale up, so as to ensure the effectiveness of the aluminum adjuvant and the quality of the vaccine. Summary of the invention

[0006] To solve the above problems, the purpose of the present invention is to provide an aluminum hydroxide adjuvant with a pseudo-boehmite structure and a preparation method and application thereof. The preparation method of the aluminum hydroxide adjuvant has reliable process, stable quality and is suitable for preventive vaccines.

[0007] In one aspect, the present invention provides a method for preparing an aluminum hydroxide adjuvant having a pseudo-boehmite structure, wherein the method comprises:

[0008] (1) Under stirring conditions, an aluminum salt solution and a sodium hydroxide solution are simultaneously pumped into a 0.05%-0.5% sodium chloride solution heated to a set temperature, and the pumping speeds of the aluminum salt solution and the sodium hydroxide solution are adjusted to control the ratio of hydroxyl groups to aluminum ions in the reaction system to be 2.5-3.5;

[0009] (2) When the aluminum ion concentration in the reaction system reaches 0.07-0.09 M, stop pumping the aluminum salt solution into the reaction system, continue pumping the sodium hydroxide solution, and stop pumping the sodium hydroxide solution until the pH of the reaction system reaches 7.8-8.5;

[0010] (3) adding 0.05%-0.5% sodium chloride solution to the prepared amount into the reaction system, performing a hydrothermal reaction, and discarding the supernatant after standing to obtain a reaction product;

[0011] (4) After the reaction product is sterilized by wet heat, the aluminum hydroxide adjuvant having a pseudo-boehmite structure is obtained.

[0012] According to a specific embodiment of the present invention, preferably, the stirring speed is 200-400 rpm, preferably 250-350 rpm.

[0013] According to a specific embodiment of the present invention, preferably, the set temperature is 40-80°C, preferably 50-60°C.

[0014] According to a specific embodiment of the present invention, preferably, in step (1), the volume of the sodium chloride solution is 45%-65%, preferably 50%-60% of the prepared amount.

[0015] According to a specific embodiment of the present invention, preferably, the preparation amount is 30-50L.

[0016] According to a specific embodiment of the present invention, preferably, the aluminum salt is aluminum chloride hexahydrate.

[0017] According to a specific embodiment of the present invention, preferably, the concentration of the aluminum chloride hexahydrate is 0.2-0.5M.

[0018] According to a specific embodiment of the present invention, preferably, the pumping speed of the aluminum salt solution is 68-83 mL / min, more preferably 72-75 mL / min.

[0019] In a specific embodiment of the present invention, when the aluminum salt solution is stopped from being pumped into the reaction system, the pH of the reaction system is 5.5-7.0. According to a specific embodiment of the present invention, preferably, the concentration of the sodium hydroxide solution is 1.0-1.4M.

[0020] According to a specific embodiment of the present invention, preferably, the pumping speed of the sodium hydroxide solution is 64-75 mL / min, more preferably 68-72 mL / min.

[0021] According to a specific embodiment of the present invention, preferably, the preparation method further comprises:

[0022] Adjuvant preparation Solution preparation: Prepare aluminum salt solution (0.2-0.5 M aluminum chloride hexahydrate), alkaline solution (1.0-1.4 M sodium hydroxide) and sodium chloride solution (0.05%-0.5%).

[0023] According to a specific embodiment of the present invention, preferably, the reaction temperature of the hydrothermal reaction is 80-130°C, preferably 100-120°C.

[0024] According to a specific embodiment of the present invention, preferably, the reaction time of the hydrothermal reaction is 2-6 h, more preferably 3-5 h.

[0025] According to a specific embodiment of the present invention, preferably, the sterilization temperature of the moist heat sterilization is 115-120° C., and the sterilization time is 15-40 min.

[0026] On the other hand, the present invention provides an aluminum hydroxide adjuvant having a pseudo-boehmite structure prepared by the above method.

[0027] According to a specific embodiment of the present invention, preferably, the particle size of the aluminum hydroxide adjuvant is 8-20 μm.

[0028] According to a specific embodiment of the present invention, preferably, the specific surface area of ​​the aluminum hydroxide adjuvant is 900-1300m 2 / kg.

[0029] On the other hand, the present invention provides an application of an aluminum hydroxide adjuvant having a pseudo-boehmite structure prepared by the above method in preparing a vaccine and / or improving the antibody level in the vaccine;

[0030] According to a specific embodiment of the present invention, preferably, the vaccine is a combined vaccine containing Hib polysaccharide antigen.

[0031] The aluminum hydroxide adjuvant prepared by the preparation method of the present invention has a small particle size distribution and a large specific surface area. In the comparative study of adjuvants, the physical properties of the adjuvant are not inferior to those of commercially available adjuvants. Due to its smaller particle size and better safety, it can increase the antibody level in the vaccine. It has broad application prospects in the development of aluminum-adjuvanted vaccines or aluminum-adjuvanted vaccine platforms, and provides more options for aluminum-adjuvanted vaccines, breaking the technical bottleneck and monopoly, and has good economic and application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 This is the particle size distribution diagram of the aluminum hydroxide adjuvant prepared in Example 1 of the present invention.

[0033] Figure 2 This is the XRD pattern of the aluminum hydroxide adjuvant prepared in Example 1 of the present invention.

[0034] Figure 3 for XRD patterns of adjuvants.

[0035] Figure 4 The appearance of the combined vaccine prepared with alumina adjuvant. Adjuvant (left), aluminum hydroxide adjuvant prepared in Example 1 of the present invention (right).

[0036] Figure 5 The aluminum hydroxide adjuvant prepared in Example 1 and Results of adjuvant-induced Hib IgG antibody detection. DETAILED DESCRIPTION

[0037] In order to have a clearer understanding of the technical features, purposes and beneficial effects of the present invention, the technical scheme of the present invention is now described in detail in conjunction with specific embodiments, and it should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of use of the present invention. In the embodiments, each raw reagent material can be obtained commercially, and the experimental method without specifying the specific conditions is a conventional method and conventional conditions well known in the art, or according to the conditions recommended by the instrument manufacturer.

[0038] Unless specifically defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the relevant art.

[0039] In some specific embodiments of the present invention, the preparation method of the aluminum hydroxide adjuvant having a pseudo-boehmite structure provided by the present invention comprises:

[0040] (1) Preparation of adjuvant solution: Prepare aluminum salt solution (0.2-0.5 M aluminum chloride hexahydrate), alkaline solution (1.0-1.4 M sodium hydroxide) and sodium chloride solution (0.05%-0.5%).

[0041] (2) Under the condition of a stirring speed of 200-400 rpm, an aluminum salt solution and a sodium hydroxide solution are simultaneously pumped into a 0.05%-0.5% sodium chloride solution heated to a set temperature of 40-80° C., the pumping speed of the aluminum salt solution is adjusted to 68-83 mL / min, and the pumping speed of the sodium hydroxide solution is adjusted to 64-75 mL / min, and the ratio of hydroxyl groups to aluminum ions in the reaction system is controlled to be 2.5-3.5; wherein the volume of the sodium chloride solution is 45%-65% of the total preparation amount, and the preparation amount is 30-50 L.

[0042] (3) When the aluminum ion concentration in the reaction system reaches 0.07-0.09 M, stop pumping the aluminum salt solution into the reaction system and continue pumping the sodium hydroxide solution. When the pH of the reaction system reaches 7.8-8.5, stop pumping the sodium hydroxide solution.

[0043] (4) Add 0.05%-0.5% sodium chloride solution to the reaction system to make up the prepared amount, and conduct a hydrothermal reaction at a temperature of 80-130° C. for 2-6 hours. After standing, discard the supernatant to obtain a reaction product.

[0044] (5) sterilizing the reaction product by wet heat at 115-120° C. for 15-40 min to obtain the aluminum hydroxide adjuvant having a pseudo-boehmite structure.

[0045] In some specific embodiments of the present invention, the preparation method of the aluminum hydroxide adjuvant having a pseudo-boehmite structure provided by the present invention comprises:

[0046] (1) Preparation of adjuvant solution: Prepare aluminum salt solution (0.2-0.5 M aluminum chloride hexahydrate), alkaline solution (1.0-1.4 M sodium hydroxide) and sodium chloride solution (0.05%-0.5%).

[0047] (2) Under the condition of a stirring speed of 250-350 rpm, an aluminum salt solution and a sodium hydroxide solution are simultaneously pumped into a 0.05%-0.5% sodium chloride solution heated to a set temperature of 50-60° C., the pumping speed of the aluminum salt solution is adjusted to 72-75 mL / min, and the pumping speed of the sodium hydroxide solution is adjusted to 68-72 mL / min, and the ratio of hydroxyl groups to aluminum ions in the reaction system is controlled to be 2.5-3.5; wherein the volume of the sodium chloride solution is 50%-60% of the total preparation amount, and the preparation amount is 30-50 L.

[0048] (3) When the aluminum ion concentration in the reaction system reaches 0.07-0.09 M, stop pumping the aluminum salt solution into the reaction system and continue pumping the sodium hydroxide solution. When the pH of the reaction system reaches 7.8-8.5, stop pumping the sodium hydroxide solution.

[0049] (4) Add 0.05%-0.5% sodium chloride solution to the reaction system to make up the prepared amount, and conduct a hydrothermal reaction at a temperature of 100-120° C. for 3-5 hours. After standing, discard the supernatant to obtain a reaction product.

[0050] (5) sterilizing the reaction product by wet heat sterilization at 115-120° C. for 15-40 min to obtain the aluminum hydroxide adjuvant having a pseudo-boehmite structure.

[0051] Example 1

[0052] (1) Add about 60% of the volume of sodium chloride solution (0.1%) into the reaction tank in advance, set the temperature to 60°C, start stirring, the speed is 300 rpm, and heat to 60°C; the preparation volume is 30L;

[0053] (2) Pumping aluminum chloride hexahydrate solution (0.4M) and NaOH solution (1.2M) at the same time, the aluminum chloride hexahydrate solution pumping rate is 75mL / min, the NaOH solution pumping rate is 70mL / min, and the ratio of hydroxyl to aluminum ion is controlled between 2.5-3.5. When the aluminum ion concentration in the system reaches 0.086M, stop pumping aluminum chloride hexahydrate solution, continue pumping NaOH solution, and stop pumping NaOH solution when the endpoint pH reaches 8.24. At this time, the ratio of hydroxyl to aluminum ion is 3.07;

[0054] (3) Add sodium chloride solution (0.1%) to the prepared amount;

[0055] (4) Perform a hydrothermal reaction, adjust the temperature to 110° C., react for 4 hours, collect in a container, let stand overnight, discard the supernatant to obtain the reaction product, and perform wet heat sterilization at 121° C. for 30 minutes to obtain an aluminum hydroxide adjuvant sample.

[0056] Example 2

[0057] (1) Add about 60% of the volume of sodium chloride solution (0.1%) into the reaction tank in advance, set the temperature to 60°C, start stirring, the speed is 300 rpm, and heat to 60°C; the preparation volume is 30L;

[0058] (2) Pumping aluminum chloride hexahydrate solution (0.4M) and NaOH solution (1.2M) at the same time, the aluminum chloride hexahydrate solution pumping rate is 75mL / min, the NaOH solution pumping rate is 70mL / min, and the ratio of hydroxyl to aluminum ion is controlled between 2.5-3.5. When the aluminum ion concentration in the system reaches 0.086M, stop pumping aluminum chloride hexahydrate solution, continue pumping NaOH, and stop pumping sodium hydroxide solution when the end point pH reaches 8.24. At this time, the ratio of hydroxyl to aluminum ion is 3.07;

[0059] (3) Add sodium chloride solution (0.1%) to the prepared amount;

[0060] (4) Perform a hydrothermal reaction, adjust the temperature to 100° C., react for 4 hours, collect in a container, let stand overnight, discard the supernatant to obtain the reaction product, and perform wet heat sterilization at 121° C. for 30 minutes to obtain an aluminum hydroxide adjuvant sample.

[0061] Comparative Example 1

[0062] (1) Add about 60% of the volume of sodium chloride solution (0.9%) into the reaction tank in advance, set the temperature to 60°C, start stirring, the speed is 300 rpm, and heat to 60°C; the preparation volume is 30L;

[0063] (2) Pumping aluminum chloride hexahydrate (0.4 M) solution into the solution in step (1) at a pumping rate of 75 mL / min, until the system temperature reaches 60° C., the total amount of aluminum chloride hexahydrate (0.4 M) solution added is 6.5 L;

[0064] (3) adding sodium hydroxide solution (1.2 M) to the solution in step (2) at a pump rate of 75 mL / min and maintaining the reaction temperature at 60° C.;

[0065] (4) When the pH of the solution in step (3) reaches 8.48, stop adding the sodium hydroxide solution;

[0066] (5) Add sodium chloride solution (0.9%) to the prepared amount;

[0067] (6) The mixture was reacted at 60°C for 4 hours, allowed to stand overnight, and the supernatant was discarded to obtain the reaction product, which was sterilized by wet heat at 121°C for 30 minutes to obtain an aluminum hydroxide adjuvant sample.

[0068] Comparative Example 2

[0069] (1) Add about 60% of the volume of sodium chloride solution (0.1%) into the reaction tank in advance, set the temperature to 60°C, start stirring, the speed is 300 rpm, and heat to 60°C; the preparation volume is 30L;

[0070] (2) Pumping aluminum chloride hexahydrate (0.4 M) solution into the solution in step (1) at a pumping rate of 75 mL / min, until the system temperature reaches 60° C., the total amount of aluminum chloride hexahydrate (0.4 M) solution added is 6.5 L;

[0071] (3) adding sodium hydroxide solution (1.2 M) to the solution in step (2) at a pump rate of 75 mL / min and maintaining the reaction temperature at 60° C.;

[0072] (4) When the pH of the solution in step (3) reaches 8.06, stop adding the sodium hydroxide solution;

[0073] (5) Add sodium chloride solution (0.1%) to the prepared amount;

[0074] (6) The mixture was reacted at 60°C for 4 hours, allowed to stand overnight, and the supernatant was discarded and then sterilized by wet heat at 121°C for 30 minutes to obtain an aluminum hydroxide adjuvant sample.

[0075] Comparative Example 3

[0076] (1) Add about 60% of the volume of sodium chloride solution (0.1%) into the reaction tank in advance, set the temperature to 60°C, start stirring, the speed is 300 rpm, and heat to 60°C; the preparation volume is 30L;

[0077] (2) Pumping aluminum chloride hexahydrate solution (0.4M) and NaOH solution (1.2M) at the same time, the aluminum chloride hexahydrate solution pumping rate is 75mL / min, the NaOH solution pumping rate is 70mL / min, and the ratio of hydroxyl to aluminum ion is controlled between 2.5-3.5. When the aluminum ion concentration in the system reaches 0.086M, stop pumping aluminum chloride hexahydrate solution, continue pumping NaOH, and stop pumping sodium hydroxide solution when the endpoint pH reaches 8.11. At this time, the ratio of hydroxyl to aluminum ion is 3.05;

[0078] (3) Add sodium chloride solution (0.1%) to the prepared amount;

[0079] (4) reacting at 60°C for 4 hours, standing overnight, discarding the supernatant to obtain the reaction product, and sterilizing it with wet heat at 121°C for 30 minutes to obtain an aluminum hydroxide adjuvant sample.

[0080] Comparative Example 4

[0081] (1) Add about 60% of the volume of sodium chloride solution (0.9%) into the reaction tank in advance, set the temperature to 60°C, start stirring, the speed is 300 rpm, and heat to 60°C; the preparation volume is 30L;

[0082] (2) Pumping aluminum chloride hexahydrate solution (0.4M) and NaOH solution (1.2M) at the same time, the aluminum chloride hexahydrate solution pumping rate is 75mL / min, and the NaOH solution pumping rate is 70mL / min, and the ratio of hydroxyl to aluminum ion is controlled between 2.5-3.5. When the aluminum ion concentration in the system reaches 0.086M, stop pumping aluminum chloride hexahydrate solution, continue pumping NaOH, and stop pumping sodium hydroxide solution when the end point pH reaches 8.2. At this time, the ratio of hydroxyl to aluminum ion is 3.13;

[0083] (3) Add sodium chloride solution (0.9%) to the prepared amount;

[0084] (4) reacting at 110°C for 4 hours, standing overnight, discarding the supernatant to obtain the reaction product, and sterilizing it with wet heat at 121°C for 30 minutes to obtain an aluminum hydroxide adjuvant sample.

[0085] Aluminum hydroxide adjuvant detection

[0086] (1) According to the general rule 3650 of Part III of the 2020 edition of the Chinese Pharmacopoeia, the aluminum hydroxide adjuvants prepared in Comparative Examples 1-4 and Examples 1-2 were tested for appearance, solubility, identification test, adsorption rate, and sedimentation rate. The test results are shown in Table 1.

[0087] Table 1 Aluminum hydroxide adjuvant test results (Pharmacopoeia)

[0088]

[0089] Summary: According to the test results in Table 1, the quality indicators such as appearance, solubility, identification test, adsorption rate and sedimentation rate of the aluminum hydroxide adjuvant prepared according to the embodiment of the present invention meet the 2020 edition of the "Chinese Pharmacopoeia" Part III General Rules 3650 Aluminum Hydroxide Adjuvant Standard.

[0090] (2) Other quality attribute tests of aluminum hydroxide adjuvant:

[0091] Malvern laser particle size analyzer 3000E was used to detect the particle size, distribution and specific surface area of ​​the aluminum adjuvant prepared in the comparative example and the embodiment;

[0092] X-ray diffraction: Take an appropriate amount of prepared aluminum hydroxide adjuvant solution in a 50mL centrifuge tube, centrifuge at 5000rpm for 10min, discard the supernatant, add deionized water to resuspend, continue centrifugation, repeat the above steps for washing 3 times, take out the precipitate, move it to a clean drying container, dry it at room temperature, and grind it into fine powder in a mortar. Place the powder sample on a glass slide and test it using an X-ray diffractometer. The instrument model is Japan's Rigaku SmartLab SE, the test range is 5-90°, the scanning speed is 2° / min, the light source is Cu-Kα rays, the tube voltage is 40kV, and the current is 40mA. The test results are shown in Table 2.

[0093] Table 2 Aluminum hydroxide adjuvant property test results

[0094]

[0095] The prepared aluminum hydroxide adjuvant was subjected to X-ray diffraction. As shown in the test results in Table 2, Comparative Examples 1 and 2 had no pseudo-boehmite structure; when the aluminum hydroxide adjuvant was added dropwise simultaneously and the ratio of Al to hydroxyl was controlled to a certain value, Comparative Examples 3, 4 and Examples 1 and 2 all showed the same structure as the commercially available aluminum hydroxide adjuvant. A similar pseudo-boehmite structure; however, the pseudo-boehmite structure of Comparative Example 3 disappeared after high-pressure sterilization, while Examples 1 and 2 maintained the pseudo-boehmite structure; Figure 1 Shown is the XRD pattern of the aluminum hydroxide adjuvant prepared in Example 1, as shown in Figure 2 Shown XRD patterns of adjuvants.

[0096] In the preparation method of the present invention, after reducing the concentration of pre-added sodium chloride, the particle size and specific surface area of ​​the aluminum hydroxide adjuvant prepared in Examples 1 and 2 are better than those in Comparative Examples 1 and 4; the particle size distribution of the aluminum hydroxide adjuvant prepared in Example 1 is as follows: Figure 3 shown.

[0097] Application of aluminum hydroxide adjuvant

[0098] The aluminum hydroxide adjuvant prepared in Example 1 was mixed with The adjuvant is used in the preparation of a combined vaccine containing polysaccharide antigens of Haemophilus influenzae type b, and the preparation method is as follows: Aluminum hydroxide adjuvant is used to adsorb Hib antigens, and then pertussis component antigen stock solution, diphtheria toxoid stock solution and tetanus toxoid stock solution are added. The Hib content in the prepared vaccine is based on the "Haemophilus influenzae type b conjugate vaccine" in Part III of the "Chinese Pharmacopoeia", which is 20ug of polysaccharide antigen per 1mL.

[0099] (1) The combined vaccine prepared with the aluminum hydroxide adjuvant prepared in Example 1 has the appearance of a milky white colloidal suspension without any clots or foreign matter that cannot be shaken off, and is more suitable for the preparation of combined vaccines containing Hib polysaccharide antigens. Figure 4 As shown, the left picture is The right picture is the appearance picture of the combined vaccine prepared with aluminum hydroxide adjuvant in Example 1 of the present invention.

[0100] (2) The combined vaccine prepared in Example 1 was used to test the serum anti-Hib IgG antibody level according to the method of "3.5.4 Efficacy Test" in "Haemophilus influenzae type b conjugate vaccine" of the Chinese Pharmacopoeia. Figure 5 As shown, the results show that compared with the commercially available Aluminum hydroxide adjuvant. The aluminum hydroxide adjuvant prepared in Example 1 of the present invention can significantly increase the level of Hib IgG antibodies in the combined vaccine.

Claims

1. A method for preparing an aluminum hydroxide adjuvant having a pseudo-boehmite structure, wherein: The method includes: (1) Under stirring conditions, an aluminum salt solution and a sodium hydroxide solution are simultaneously pumped into a 0.05%-0.5% sodium chloride solution heated to a set temperature, and the pumping speeds of the aluminum salt solution and the sodium hydroxide solution are adjusted to control the ratio of hydroxyl groups to aluminum ions in the reaction system to be 2.5-3.5; (2) When the aluminum ion concentration in the reaction system reaches 0.07-0.09 M, stop pumping the aluminum salt solution into the reaction system, continue pumping the sodium hydroxide solution, and stop pumping the sodium hydroxide solution until the pH of the reaction system reaches 7.8-8.5; (3) adding 0.05%-0.5% sodium chloride solution to the prepared amount into the reaction system, performing a hydrothermal reaction, and discarding the supernatant after standing to obtain a reaction product; (4) After the reaction product is sterilized by wet heat, the aluminum hydroxide adjuvant having a pseudo-boehmite structure is obtained.

2. The preparation method according to claim 1, wherein The stirring speed is 200-400 rpm, preferably 250-350 rpm.

3. The preparation method according to claim 1, wherein The set temperature is 40-80°C, preferably 50-60°C.

4. The preparation method according to claim 1, wherein In step (1), the volume of the sodium chloride solution is 45%-65% of the prepared amount, preferably 50%-60%; Preferably, the preparation volume is 30-50L.

5. The preparation method according to claim 1, wherein The aluminum salt is aluminum chloride hexahydrate; Preferably, the concentration of the aluminum chloride hexahydrate is 0.2-0.5M.

6. The preparation method according to claim 1, wherein The aluminum salt solution is pumped at a speed of 68-83 mL / min, preferably 72-75 mL / min.

7. The preparation method according to claim 1, wherein The concentration of the sodium hydroxide solution is 1.0-1.4M; Preferably, the pumping speed of the sodium hydroxide solution is 64-75 mL / min, more preferably 68-72 mL / min.

8. The preparation method according to claim 1, wherein The reaction temperature of the hydrothermal reaction is 80-130°C, preferably 100-120°C; Preferably, the reaction time of the hydrothermal reaction is 2-6 h, more preferably 3-5 h.

9. Aluminum hydroxide adjuvant with pseudo-boehmite structure prepared by the method according to any one of claims 1 to 8; Preferably, the particle size of the aluminum hydroxide adjuvant is 8-20 μm; Preferably, the specific surface area of ​​the aluminum hydroxide adjuvant is 900-1300m 2 / kg.

10. Use of the aluminum hydroxide adjuvant having a pseudo-boehmite structure according to claim 9 in preparing vaccines and / or improving antibody levels in vaccines; Preferably, the vaccine is a combination vaccine containing Hib polysaccharide antigen.

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