Conjugate of hapten and amino-containing carrier and preparation method and usage thereof

A hapten and amino group-containing technology, which is applied in the direction of carrier-binding antigen/hapten components, anti-inflammatory agents, biological testing, etc., can solve the problems of limited targeted immunity and failure to prevent and treat chronic infections, and achieve enhanced specific immunity, Improving the effect of the local immune microenvironment

Inactive Publication Date: 2010-10-13
复旦大学附属华东医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, practice has shown that the targeted immunity induced by monoclonal antibodies is generally lim

Method used

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  • Conjugate of hapten and amino-containing carrier and preparation method and usage thereof
  • Conjugate of hapten and amino-containing carrier and preparation method and usage thereof
  • Conjugate of hapten and amino-containing carrier and preparation method and usage thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039]Using the hapten dinitrofluorobenzene and anti-hepatitis B surface antigen monoclonal antibody as reaction materials, under weakly alkaline conditions, through Sanger's reaction, the fluorine falls off the hydrogen on the amino group in the monoclonal antibody, and the dinitrobenzene Partially replace and covalently bind to the amino group to obtain the target product dinitrobenzene covalently bound to the hepatitis B surface antigen monoclonal antibody, one of the target products, the binding rate: 1-200 dinitrobenzene, the binding amount is not Affect the specific recognition function of monoclonal antibody.

[0040] Dinitrochlorobenzene, dinitrobromobenzene, or dinitroiodobenzene may also be used as the above-mentioned hapten.

[0041] After immunoassay (such as ELISA), the results show that the signal of the immunoreaction assay is amplified up to 200 times, which significantly improves the sensitivity of the immunoassay.

[0042] Mix the aqueous solution of HBIG (1...

Embodiment 2

[0045] According to the preparation method of Example 1, a tumor or infection targeting agent covalently bound by dinitrobenzene is used to prepare a hapten-induced immunotargeted therapy tumor drug, or / and directly use dinitrofluorobenzene as the original drug It has anticancer effect. The results showed that the binding ratio was the same as that in Example 1; the tumor inhibition rate of subcutaneously transplanted tumors in mice injected with DNFB was good.

[0046] The tumor or infection targeting preparation can be selected from anti-HER-2 monoclonal antibody Herceptin or hepatitis B monoclonal antibody.

Embodiment 3

[0048] According to the preparation method of Example 1, the immune antigen or epitope of the tumor or chronic infection pathogen covalently bonded with dinitrobenzene is used to prepare the prevention and treatment drug induced by hapten or to strengthen specific immunity. For example, DNFB combined with hepatitis B surface antigen, DNFB combined with melanoma tumor regression antigen epitope, etc., can be used for specific prevention or as a tumor therapeutic vaccine, etc. The results show that the binding ratio is the same as that of Example 1. The drug for targeted treatment of tumor or chronic infection is characterized in that the compound with targeting effect is used as the guiding part for binding tumor, and is covalently combined with the hapten determinant part to carry out targeted treatment of tumor or chronic infection.

[0049] Table 1: 48-hour cell viability of SMMC-7721 liver cancer cell line in vitro (MTT method, mean value of six wells).

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Abstract

The invention belongs to the field of drug synthesis, and relates to a combined conjugate of a hapten and an amino-containing carrier, and a preparation method as well as the usage thereof. In the invention, the combined conjugate of the hapton and the amino-containing carrier is obtained via Sanger's reaction. A signal can be amplified during micro determination by the immunity of the hapten and the function of the carrier which are combined with the monoclonal antibody for the hapten, and the sensitivity of determination is obviously improved. Amino-containing dinitrobenzol structure is taken as a war head with a killing function when inducing cellular immunity in vivo, is combined with the carrier with a targeting function, has peculiar discrimination on cancer and chronic infection, locally induces multiple immunity activating factors at targeting lesion parts, improves local immunity micro environment, and treats cancer or infection by strengthening immunization. The conjugate can be used for micro determination of immunization and the preparation of drug for preventing and curing tumor.

Description

technical field [0001] The invention belongs to the field of pharmaceutical preparations, and relates to a combination of a hapten and an amino-containing carrier, a preparation method and application thereof. Background technique [0002] It is known that haptens are incomplete single antigens, which can be combined with proteins after external application on the skin of the body or oral administration of animals, and induce a strong cellular immune response in the body about 7-10 days later. Haptens such as dinitrofluorobenzene and Dinitrofluorobenzene is not naturally produced in organisms, and in all organisms in nature, the amino acid residues of proteins do not contain dinitrobenzene structures, unless artificially marked. Some studies have disclosed that through the Sanger's reaction, that is, the dinitrobenzene of DNFB or DNCB replaces the hydrogen on the amino group, and under slightly alkaline conditions, the protein is mixed with DNFB to achieve binding. The meth...

Claims

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Application Information

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IPC IPC(8): A61K39/385A61P35/00A61P29/00G01N33/53
Inventor 戴兆云储艳秋何晓俊匡春香黄伟达蒋公羽方明珠
Owner 复旦大学附属华东医院
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