Derivatives of nucleosides, processes for the production of thes

Details Derivatives of nucleosides, processes for the production of thes Derivatives of nucleosides, processes for the production of thes

1996-06-05

1998-02-24

Huff, Sheela

Chemistry: natural resins or derivatives; peptides or proteins;

Proteins, i.e., more than 100 amino acid residues

Blood proteins or globulins, e.g., proteoglycans, platelet...

530300, 530350, 530324, 5303881, 5303889, 530362, 530363, 53038821, 5303898, 5303891, 544118, 544123, 435333, C07K 1600, C07K 500, C07K 700, C12P 2108

Patent

active

057213410

DESCRIPTION:

BRIEF SUMMARY
The invention relates to derivatives of nucleosides, processes for the production of these derivatives of nucleosides, as well as specific polyclonal and monoclonal antibodies of the aforementioned derivatives.
The macromolecule DNA (deoxyribonucleic acid) is the constituent of chromosomes and different segments of this molecule form genes, which are the supports of hereditary characteristics. DNA is in the form of a double spiraled helix formed alternatively by sugar (deoxyribose) and phosphate, the spirals of the two chains being locally joined by groups of purine or pyrimidine-type, nitrogen nucleic bases. The nucleotides constituting DNA are phosphoric esters of nucleosides.
The nucleic bases of DNA of an individual (or an animal or vegetable/plant) can be modified and damaged when said individual is exposed to intense solar radiation, cosmic radiation (intercontinental flights), photosensitizers, contact with asbestos or ionizing radiation, no matter whether the latter is accidental or due to a radiotherapy treatment. These modifications of the nucleic bases of DNA can lead to a significant change to the genetic patrimony of the individual in question. It is particularly important to detect if such modifications are produced and to define the nature of the modifications which have occurred.
It would therefore be of interest to develop a determination or assaying procedure usable in patients who may have undergone DNA modifications.
Among the various assaying procedures making it possible to determine the presence of modified DNA in a sample, immunoassays consist of reacting a specific antibody of a particular DNA modification with a sample containing isolated or hydrolyzed DNA. These antibodies are generally produced by cloning.
The production of polyclonal or monoclonal antibodies firstly requires the production of specific antigens, i.e. nucleosides or nucleotides, whose nucleic bases have undergone the modifications which it is wished to detect, said nucleotides or nucleotides being connected to a large molecule, e.g. a protein. A nucleoside or nucleotide alone would be too small to be seen by the immune system. The polyclonal antibodies are then produced by a mammal which has received an injection of the said antigen, which contains a protein foreign to the mammal in question, linked with a hapten (small molecule against which it is wished to obtain the specific antibodies).
The prior art discloses methods for the immunological determination of nucleic acids. The article by Christopher P. WILD: "Antibodies to DNA alkylation adducts as analytical tools in chemical carcinogenesis", Mut. Res., 1990, 233, pp 219-233, is devoted to specific antibodies of nucleotides, whose nucleic bases have undergone modifications by alkylation. This author insists on the function of the antibodies in the immunological determinations used in epidemiological studies of human cancers and chemical carcinogenesis, due in particular to alkylating agents.
The article by B D. Stollar: "Immunochemical analyses of nucleic acids", Progress in Nucleic Acid, Research and Molecular Biology, 1992, 42, pp 39-75 also relates to the specific antibodies of nucleic acids.
Certain oxidative deficiencies of DNA have also formed the subject matter of various publications.
Thus, the article by G. J. West et al: "Radioimmunoassay of 8-hydroxyadenine" Int. J. Rad. Biol., 1982, 42, pp 481-490 describes radioimmunoassays of 8-hydroxyadenine. This DNA modification can arise following irradiation with rays.
The article by P. Degan et al "Quantitation of 8-hydroxy-2'-deoxyguanosine in DNA by polyclonalantibodies", Carcinogenesis, 1991, 12, pp 865-871 describes specific polyclonal antibodies of 8-hydroxy-2'-deoxyguanosine and 8-hydroguanine. These polyclonal antibodies can be used in immunoassays in order to rapidly isolate the two aforementioned types of modified guanosine, e.g. in a urine sample.
The article by H. L. Lewis et al "Serologic assay of DNA base damage", Rad. Res. 1978, 75 pp 305-316 describes the preparation of polyclonal antibodies

REFERENCES:
patent: 5382580 (1995-01-01), Chen et al.
Erlanger et al PNAS, USA vol. 52 p. 68, Jul. 1964.
Degan et al, Carcinogensis vol. 12 p. 865, 1991.

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