Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1991-01-03
1993-12-21
Ceperley, Mary E.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
514185, 536 2533, C12Q 168
Patent
active
052720560
DESCRIPTION:
BRIEF SUMMARY
,15 N? 14?
A kit is also provided for a nucleic acid assay for locating guanine groups in nucleic acids, oligonucleotides, polynucleotides, RNA and single-stranded DNA, and non-base paired guanine groups in double-stranded DNA. The kit comprises a polyaza metal complex selected from Structures I-III and their derivatives, an oxidant selected from the group consisting of peracid, hypochlorite, O.sub.2, peroxide in combination with ascorbate, O.sub.2 in combination with ascorbate, and a base of piperidine, N-butylamine or sodium hydroxide. The detection, for example, of labelled 5'-ends in base-cleaved oligonucleotide indicates the position of guanine groups in the oligonucleotide.
A method is also provided for treating neoplastic growth comprising administering to a subject having a neoplastic growth, an effective amount of a polyaza metal complex of Structures I-III and their derivatives.
Advantageously, the process provides a new method for guanine-specific modification of DNA in sequencing technology and in therapeutics. In addition, the currently used chemical sequencing methods use volatile and toxic chemicals such as dimethylsulfate. The invention employs less toxic materials which are more easily handled.
For a better understanding of the present invention, together with other and further objects, reference is made to the following description, taken together with the accompanying drawings and its scope will be pointed out in the appended claims.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is an autoradiogram of an electrophoresis of a polyacrylamide gel showing oligonucleotide cleavage products obtained using structure 1 with an oxidant and base treatment compared with control studies;
FIG. 2 is an autoradiogram of an electrophoresis of a polyacrylamide gel comparing efficacy of various metal complexes in the cleavage of the oligonucleotide.
DETAILED DESCRIPTION OF THE INVENTION
Polyaza Compounds
The compounds useful in the invention are characterized in one regard by their ability to form stable complexes with transition metals. Stable complexes are those having measurable lifetimes at room temperature in water or common organic solvents.
The compounds comprise at least one ligand and a transition metal. A ligand is defined herein as a molecule that is attached to the central metal atom of a coordination compound. Preferred ligands are tetradentate or pentadentate and may comprise either macrocyclic or non-macrocyclic molecules. Other ligands, however, can also be used. It is to be understood that a tetradentate ligand has four donor atoms while a pentadentate ligand has five donor atoms.
The donor atoms may be nitrogen, oxygen, and/or sulfur. Preferably at least two donor atoms are nitrogens, which are separated by from two to four carbon atoms.
Suitable nitrogen donor groups are the amino group of peptides, or amines, imines, pyridines, imidazoles, pyrroles, and pyrazoles, with imine and pyridine groups preferred. The configuration may be square planar or pyramidal, with square planar preferred, but is not limited to these. Suitable oxygen donor groups are phenol, alcohol, carboxylic acid, and carbonyl. Examples of molecules containing oxygen donor groups are salens, salophens and crown thio ethers. Generally, crown ethers containing only oxygen donor groups are not suitable herein. Sulfur donor groups may be, e.g. thiols, thiolates and thiophenols. Complexing molecules containing sulfur groups may contain only sulfur donor groups such as in the crown thio ethers, or the sulfur groups may be combined with oxygen and/or nitrogen donor groups in a suitable ligand.
The ability of the ligands to form stable complexes results from the relative positions of the donor groups. Much of the rest of the ligand consists of carbon atoms that may be thought of as collectively forming a scaffold for maintaining the proper position of the donor groups.
Substituents on the atoms of the ligand affect the properties of the compounds, such as their ability to bind the nucleotides and DNA, their effectiveness i
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Wagle
This invention was made with government support under grant number CHE 9006684 awarded by the National Science Foundation. The government has certain rights in the invention.
The invention relates to the modification of nucleic acids by oxidation using metal complexes of polyaza ligands.
The technique of irreversible DNA modification holds great potential as an in vitro tool for molecular biologists. Such modification raises the possibility of DNA - directed drug therapy in vivo. Currently prescribed chemotherapeutic agents acting at the level of DNA are often effective, but their therapeutic index is poor, limited by the lack of target specificity.
Naturally-occurring and laboratory-designed agents for DNA modification have often relied on transition metal ions for nu
Burrows Cynthia J.
Chen Xiaoying
Rokita Steven E.
Ceperley Mary E.
The Research Foundation of State University of New York
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