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Aldosterone derivatives, immunogens, and synthesis methods, specific antibodies, detection reagents, preparation methods, and kits

An aldosterone and derivative technology, applied in biological testing, immunoglobulins, chemical instruments and methods, etc., can solve the problems of unsuitability for large-scale clinical sample determination, high testing cost, poor stability, etc., to meet the needs of clinical testing, High sensitivity and accurate results

Active Publication Date: 2019-08-06
苏州博源医疗科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the existing aldosterone detection methods have certain limitations, such as complex operation, poor stability, need for special equipment, high detection cost, etc., and are not suitable for the determination of large quantities of clinical samples
At present, there is a lack of aldosterone detection reagents with good stability, high sensitivity and strong specificity in the market, especially good quality automatic detection reagents

Method used

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  • Aldosterone derivatives, immunogens, and synthesis methods, specific antibodies, detection reagents, preparation methods, and kits
  • Aldosterone derivatives, immunogens, and synthesis methods, specific antibodies, detection reagents, preparation methods, and kits
  • Aldosterone derivatives, immunogens, and synthesis methods, specific antibodies, detection reagents, preparation methods, and kits

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preparation example Construction

[0058] The synthetic method of aldosterone derivative comprises:

[0059] The carboxyl hydroxyl group of aldosterone is halogenated to obtain a halide. Using the halide as the substrate, the primary amine halogenation reaction with tert-butyl 4-aminobutyrate gives intermediate products containing secondary amine groups. After protecting the secondary amine group in the intermediate product, hydrolysis of the ester group occurs under acidic conditions.

[0060] In the halogenation reaction of the carboxyl hydroxyl group of aldosterone, the halogenation reagents used in the halogenation reaction are lithium chloride (LiCL) and methanesulfonyl halide (MsCL). Aldosterone first reacts with MsCL, and Ms- replaces the hydroxyl hydrogen in the carboxyhydroxyl group of aldosterone. Then, the chlorination reaction with LiCL was carried out.

[0061] When protecting the secondary amine group in the intermediate product, Boc anhydride is used as the protecting agent in the embodiment o...

Embodiment 1

[0128] 1. Synthesis of aldosterone derivatives.

[0129] The synthetic route of aldosterone derivatives is as follows figure 1 shown.

[0130] Wherein, compound 2 is synthesized as follows.

[0131]

[0132] Under the condition of 10°C, 55 grams of catalyst (Cbz-Cl, benzyl chloroformate) was added at one time in the sodium hydroxide solution of 2N normality (2mol / L) dissolved with 30 grams of compound 1 (4-aminobutyric acid) . The mixture was stirred overnight at room temperature. During this period, the reaction process was monitored by TLC until the reaction was complete. The reacted mixture was extracted with ethyl acetate (3x300 mL, 3 times, 300 mL each time). The aqueous layer was adjusted to pH=3 with 1N normal hydrochloric acid and extracted with dichloromethane (3x300 mL). The organic layers were combined, washed with water (200 mL), brine (200 mL), dried in vacuo, and concentrated to obtain compound 2 (33 g, yield 48.8%) as a white solid.

[0133] Wherein, c...

Embodiment 2

[0155] Synthesis of the aldosterone immunogen.

[0156] The aldosterone immunogen is composed of bovine serum albumin (Bovine Serum Albumin, BSA) and -NH-(CH 2 ) 3 The -CO- group is connected. The specific steps of the synthetic method of aldosterone immunogen are as follows:

[0157] 1. Dissolve 200mg of bovine serum albumin in 50ml of 0.2M, pH 8.5 phosphate buffer.

[0158] 2. Add the following chemicals into a small beaker and stir to dissolve: 200mg of the aldosterone derivative synthesized in Example 1, 3.5ml of dimethylformamide, 3.5ml of ethanol, 7.0ml of 10mM potassium phosphate buffer at pH 5.0, 200mg of 1 -Ethyl-3-(-3-dimethylaminopropyl) carbodiimide, 50 mg of N-hydroxysulfosuccinimide, these chemicals were stirred and dissolved at room temperature for 30 min.

[0159] 3. The dissolved solution was added dropwise to the BSA solution, and stirred overnight at 2-8° C. to obtain the antigen; the synthesized antigen was purified by dialysis to obtain the aldosterone...

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Abstract

The invention discloses an aldosterone derivative and an aldosterone immunogen, synthesis methods thereof, a specific antibody, an aldosterone detection reagent, a preparation method of the detection reagent, and a kit, and belongs to the biotechnical field. The aldosterone immunogen has high immunogenicity, can be induced to obtain the high-tilter anti-aldosterone specific antibody, and has no cross reaction with 92 common interferents; and the aldosterone detection reagent prepared from the antibody can accurately and rapidly determine the content of aldosterone in a sample. Compared with existing commercial detection reagents, the detection reagent in the invention has the advantages of simplicity in operation, high sensitivity, high specificity, accuracy in result, effective reduction of the aldosterone detection cost, and facilitation of clinic large-scale promotion and use.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an aldosterone derivative, an immunogen, a synthesis method, a specific antibody, a detection reagent, a preparation method, and a kit. Background technique [0002] Aldosterone (Aldosterone), its structural formula is shown in the following formula (III). [0003] [0004] Aldosterone is a mineralocorticoid synthesized and secreted by the zona glomerular cells of the adrenal cortex. Aldosterone mainly acts on the renal distal convoluted tubule and renal cortical collecting duct, which can increase the reabsorption of sodium ions and promote the excretion of potassium ions. In addition, aldosterone can also act on the medullary collecting duct, promote the excretion of hydrogen ions, and acidify the urine. Aldosterone is secreted through the renin-angiotensin-aldosterone system. When the volume of extracellular fluid decreases, the cells next to the glomerulus are stimulated to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07J41/00C07K14/765C07K14/795C07K14/47C07K16/44G01N33/64G01N33/577
CPCC07J41/005C07K14/47C07K14/765C07K14/795C07K16/44C07K19/00G01N33/577G01N33/64
Inventor 虞留明程黎明沙德顺
Owner 苏州博源医疗科技有限公司
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