Antigen and preparation method and application thereof

An antigen and reaction technology, applied in chemical instruments and methods, carrier-binding antigen/hapten components, pharmaceutical formulations, etc., can solve the problems of patients' negative emotions, hinder endogenous opioids, etc., and prevent relapse. Effect

Active Publication Date: 2011-04-06
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these antagonists can be used to treat heroin addiction, their use is limited because both antagonists are inactive in the absence of exogenous opioids and can also simultaneously block Endogenous opioids, which contribute to negative mood in chronically treated patients

Method used

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  • Antigen and preparation method and application thereof
  • Antigen and preparation method and application thereof
  • Antigen and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, the preparation of morphine-6-glutaric acid monoester-bovine serum albumin antigen (morph-BSA)

[0047] Small molecule (small molecule less than 10KDa) drugs must be coupled to a carrier protein before they can cause an immune response. Therefore, in order to generate an immune response, the synthesis of small molecule drug derivatives, that is, the synthesis of haptens, must first be carried out.

[0048] Antigen synthesis route diagram as figure 1As shown, the specific steps are as follows:

[0049] 1. Preparation of morphine-6-glutaric acid monoester

[0050] (1) Dissolve 1.3g of morphine hydrochloride in 30mL of water, add dropwise ammonia water to pH 8.5, react at a temperature of 37°C with stirring for 10min, filter with suction to obtain an off-white solid, and dry it in vacuum for 24h to obtain 1g of free morphine alkali.

[0051] (2) Mix 1 g of morphine free base and 1.2 g of glutaric anhydride obtained in step (1) with a catalytic amount of N...

Embodiment 2

[0061] Embodiment 2, the preparation of morphine-6-glutaric acid monoester-hemocyanin antigen (morph-KLH)

[0062] Antigen synthesis route diagram as figure 1 As shown, the specific steps are as follows:

[0063] 1. Preparation of morphine-6-glutaric acid monoester

[0064] Same as Step 1 of Experiment 1.

[0065] 2. Coupling of morphine-6-glutaric acid monoester and carrier protein

[0066] (1) Dissolve 100 mg of morphine-6-glutaric acid monoester obtained in step 1 and 20 mg of hemocyanin in TE buffer solution (pH 8.0), and fully oscillate to dissolve to obtain liquid I;

[0067] (2) Take 48mg of 1-ethyl-3-3 dimethylaminopropyl carboimide (1-ethyl-3-(3-dimethylaminopropyl), EDC) and fully dissolve in TE buffer solution (pH 8.0) to obtain II liquid;

[0068] (3) Add liquid I dropwise to liquid II while shaking, and react in a shaker at 37°C for 2 hours to obtain a conjugate formed by covalently linking morphine-6-glutaric acid monoester and hemocyanin; The covalent bond...

Embodiment 3

[0070] Embodiment 3, the application of antigen

[0071] 1. Actively immunized mice and serum antibody titer analysis

[0072] 1. Prepare antibody using morph-KLH antigen

[0073] Take BABL / C mice (20-25g) as experimental animals. The mice were raised in cages with controllable temperature (21° C.-24° C.) and lights (08:00 on and 20:00 off), free to eat and drink for 5 days, with a body weight of about 25-28 g. Randomly divided into three groups, each group of 8 mice, the three groups of mice were subjected to the following treatments: A, injection of adjuvant+KLH; B, injection of adjuvant+morph-KLH (50 μg / animal / 0.2ml, that is, each Only mice are injected with the mixture of 0.2ml adjuvant and morph-KLH, containing 50 μ g morph-KLH in the 0.2 ml mixture); Adjuvant + moroh-KLH, 0.2ml injection contains 100 μg morph-KLH). The injection method is subcutaneous injection, the adjuvant is Freund's complete adjuvant, and the drug is divided into four injection points on average....

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Abstract

The invention discloses an antigen and a preparation method and application thereof. The antigen is a conjugate formed by connecting morphine-6-monoglutarate and a carrier protein through a covalent bond. The method for preparing the antigen comprises the following step of: conjugating the morphine-6-monoglutarate with the carrier protein so as to prepare the conjugate of the morphine-6-monoglutarate and the carrier protein, wherein the carrier protein is bovine serum albumin or hemocyanin. The antigen can stimulate an organism to generate a morphine antibody and specifically recognize the morphine and heroin. Mouse experiments prove that: a vaccine which takes the antibody as an active substance can prevent morphine relapse to a certain degree.

Description

technical field [0001] The present invention relates to an antigen and its preparation method and application. Background technique [0002] Drug addiction is a major public health and social problem that plagues global human health and social development. It seriously endangers human health, spreads diseases, affects family stability and social development, and causes huge economic losses to society. According to the 2006 report of the United Nations Office on Drugs and Crime, there are 224 million drug abusers in the world, and the annual economic loss is 800 billion US dollars. The drug problem in our country has been resurgent since the 1980s, and the situation has always been very serious. By the end of 2005, the number of registered drug addicts had reached 1.16 million. Moreover, according to statistics, in 2007, 42% of the newly-infected cases of AIDS in my country were caused by virus infection and transmission caused by drug users sharing syringes, causing major p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D489/02C07K14/765C07K14/77C07K14/795C07K14/47C07K14/75C07K14/33A61K39/385A61K47/48A61P25/36A61K47/64
Inventor 陆林李芊芊
Owner PEKING UNIV
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