Cyclodextrin esterified derivative, preparation method and application of cyclodextrin esterified derivative

A technology of cyclodextrin and derivatives, which is applied in the direction of non-active ingredients of polymer compounds, pharmaceutical formulas, and medical preparations containing active ingredients, etc., can solve the problem that the quality requirements of veterinary vaccines cannot be met, and it is difficult to produce veterinary vaccines. Wide application, weak immune activity and other problems, achieve good thermal stability and centrifugal stability, improve antigen immune activity, and simple method

CN104086676AActive Publication Date: 2014-10-08JIANGSU ACAD OF AGRI SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2014-10-08

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Abstract

The invention relates to a cyclodextrin esterified derivative, a preparation method and an application of the cyclodextrin esterified derivative, belonging to the technical field of oil in water adjuvants. The cyclodextrin esterified derivative is obtained by esterifying cyclodextrin by lauroyl chloride and sulfuric acid. The invention further provides a preparation method of the cyclodextrin esterified derivative. The preparation method is characterized by comprising the following steps: (1) carrying out esterification reaction on cyclodextrin and acyl chloride; (2) carrying out esterification reaction on a mixture obtained after the reaction in the step (1) and chlorosulfonic acid; and (3) taking the mixture obtained after the reaction in the step (2), adjusting the pH to 6.8-7.2, removing the solvent, and dissolving, dialyzing and drying the precipitate to obtain the cyclodextrin esterified derivative. The invention further provides an oil in water adjuvant and a veterinary vaccine. The oil in water adjuvant prepared by the cyclodextrin esterified derivative has good stability and centrifugal stability, and the immunocompetence of the antigen can be remarkably improved. The preparation method of the cyclodextrin esterified derivative provided by the invention is simple, low in degree of substitution of sulfate radicals, high in degree of substitution of lauroyl and high in yield.
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Description

technical field

[0001] The invention belongs to the technical field of oil-in-water adjuvants, and in particular relates to a cyclodextrin esterified derivative, a preparation method and an application thereof. Background technique

[0002] There are currently three formulations of oil emulsion vaccines: water-in-oil (W / O), water-in-oil-in-water (W / O / W) and oil-in-water (O / W). W / O dosage form has the strongest immune activity, but has severe side effects and difficult injection; W / O / W dosage form also has strong immune activity, but has poor thermal stability and short shelf life; O / W dosage form has less toxic and side effects, is convenient to use and heat Good stability, but weak immune activity. MF59 is the first O / W dosage form adjuvant to be commercially applied, and the human influenza vaccine prepared with it has been marketed in Europe and the United States. The oil of the adjuvant is squalene, which has the advantages of safety, minimal toxic and side effects, et...

Examples

Embodiment 1

[0024] Example 1 Preparation of cyclodextrin esterified derivatives

[0025] (1) Materials and instruments

[0026] Test drugs: cyclodextrin (molecular weight 1135, β type, Shanghai Kaiyang Biotechnology Co., Ltd., batch number T0008), N,N-dimethylformamide, pyridine, sulfuric acid, hydrochloric acid, chlorosulfonic acid, ammonia water (analytical grade, Nanjing Chemical Reagent Co., Ltd.); lauroyl chloride (aladdin industrial corporation, batch number F1228036); potassium chromate, sodium chloride, and anhydrous sodium sulfate (Xilong Chemical Co., Ltd.).

[0027] Main equipment: DZF-6051 vacuum drying oven (Shanghai Qixin Scientific Instrument Co., Ltd.); SHI-82A constant temperature water bath oscillator (Changzhou Guohua Electric Co., Ltd.); rotary evaporator (Shanghai Yarong Biochemical Instrument Factory); SHZ- Ⅲ Circulating water vacuum pump (Shanghai Yarong Biochemical Instrument Factory); KH-500DE CNC ultrasonic cleaner (Kunshan Hechuang Ultrasonic Instrument Factory...

Embodiment 2

[0059] Example 2 Preparation of O / W Emulsion by Esterification Derivatives of Cyclodextrin

[0060] (1) Preparation method

[0061] Get 6 cyclodextrin esterification derivatives (SLP) with higher degree of substitution of lauroyl group in Example 1, i.e. cyclodextrin esterification derivatives prepared by test numbers 2, 4, 5, 6, 7, 8, respectively The corresponding oil-in-water emulsions A-SLP2, A-SLP4, A-SLP5, A-SLP6, A-SLP7 and A-SLP8 were prepared.

[0062] Preparation method of oil-in-water emulsion: Take 1.0g of cyclodextrin esterified derivatives, dissolve in 2.0g of Tween-80, and emulsify to prepare the Tween-80 solution of SLP; slowly add the Tween-80 solution of SLP Add 80ml of water for injection to make a transparent or slightly opalescent aqueous phase solution; add 8.0g of white oil to the aqueous phase solution, fill it up with water for injection to a final volume of 100ml, and obtain an oil-water mixture; homogenize the oil-water mixture with high pressure M...

Embodiment 3

[0068] Example 3 O / W Adjuvant Prepared by Cyclodextrin Esterification Derivatives Study on the Immunological Activity of Mice

[0069] (1) Mixing method: PCV-2 (porcine circovirus type 2) antigen solution (ZJ / C strain, virus content before inactivation is 10 7.0 TCID 50 / ml) were mixed with the oil-in-water emulsion A-SLP8 and ISA15A VG in Example 2 in a volume ratio of 1:1 to prepare test vaccine and control vaccine 1; Mix 1:2.5 evenly to prepare the control vaccine 2, and store it at 4°C for later use.

[0070] (2) Main reagents: Porcine circovirus type 2 ELISA antibody detection kit (Wuhan Keqian Animal Biological Products Co., Ltd.

[0071] (3) Main instruments: AH100D high-pressure homogenizer (ATS Industrial Systems Co., Ltd.); SatFax-2100 enzyme-linked immunoassay instrument (Simon Corporation, USA).

[0072] (4) Animal grouping and treatment: 40 5-week-old Balb / c female mice (purchased from the Veterinary Medicine Center of Yangzhou University) were randomly divided...