Organic compounds -- part of the class 532-570 series – Organic compounds – Carboxylic acids and salts thereof
Reexamination Certificate
2000-01-11
2001-12-11
Killos, Paul J. (Department: 1623)
Organic compounds -- part of the class 532-570 series
Organic compounds
Carboxylic acids and salts thereof
C562S433000, C562S426000
Reexamination Certificate
active
06329546
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to new caged amino acids.
2. Related Background Art
For investigating the mechanism of action of a physiologically active amino acid compound such as glutamic acid in an organism, it is necessary to quantitatively measure the rise and fall of this compound within a biological system in a short period of time. Further, it is important to observe various changes following the compound introduced into the system.
On the other hand, biological reactions are mostly very fast, and a plurality of reactions usually progress at the same time while complicatedly relating to each other. Therefore, when the amino acid compound is added from the outside, the step of its diffusing within the system rather determines the rate, whereby the subsequent reaction to be determined in practice may not be grasped clearly.
SUMMARY OF THE INVENTION
For overcoming such a problem, various methods have been proposed as a method of rapidly adding an amino acid compound as a target material. As one of such methods, a method using a technique based on irradiation with light, i.e., so-called caged reagent, has been reported. In general, this method comprises the steps of introducing into a biological system a caged compound (which refers to a compound in which a caging group is introduced or a compound labeled with a caging group) in which a specific protective group protects an active part of a physiologically active substance to be traced; verifying that this substance has sufficiently diffused to a point of application; and then liberating the protective group (caging group) upon irradiation with light, so as to release the amino acid compound, thereby making it possible to trace the target reaction caused by the amino acid compound. This caged compound (caged amino acid) is characterized in that it can release the amino acid upon irradiation with light alone, it can release the protective group very fast, and it enables a position-specific supply of the amino acid by narrowing the light irradiation only to a specific part as necessary. The present invention provides amino acid compounds having functions mentioned above, which are caged by a caging group having a novel structure.
As a result of diligent studies, the inventor has succeeded in synthesizing caged amino acids having such excellent functions, thus accomplishing the present invention.
Namely, the caged amino acids in accordance with the present invention comprise, as a basic structure, an amide bond composed of an amino group of a target amino acid compound and a carboxyl group of a cinnamic acid derivative having a specific structure. Further, in the cinnamic acid derivative, each of its aromatic ring and carboxyl group is in a trans orientation with respect to its double bond. Also, the aromatic ring is characterized in that an amino group is disposed as a substitute at an ortho position with respect to the position where a double-bonded carbon is bound.
While the new caged amino acids in accordance with the is present invention stably exist in the dark, the amide bond is severed upon an intramolecular reaction in the presence of light, whereby the amino acid compound is released.
More specifically, a new caged amino acid in accordance with the present invention is a caged amino acid expressed by the following formula 1:
where X and Y independently represent one kind selected from the group consisting of a hydrogen atom, a halogen atom, an alkyl group having a carbon number from 1 to 4, an alkyloxy group having a carbon number from 1 to 4, an alkylamino group having a carbon number from 1 to 4, and a benzo group, and may be identical or different from each other; R
1
represents one kind selected from the group consisting of a hydrogen atom and an alkyl group having a carbon number from 1 to 4; R
2
and R
3
independently represent one kind selected from the group consisting of a hydrogen atom and an alkyl group having a carbon number from 1 to 4, and may be identical or different from each other; A represents an amino acid residue selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, &ggr;-aminobutanoic acid, N-methyl-D-aspartic acid, asparagine, glutamine, lysine, hydroxylysine, arginine, cysteine, cystine, methionine, phenylalanine, tyrosine, tryptophan, histidine, proline, and 4-hydroxyproline; and M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
As preferable examples of new caged amino acids in accordance with the present invention, caged amino acids having respective structures expressed by the following formula 2 to 18 are specifically listed:
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
where M represents one kind selected from the group consisting of a hydrogen atom, an alkali metal, and an alkaline-earth metal.
As shown in
FIG. 1
, while such a cage amino acid compound stably exists in the dark since the double bond of the cinnamate group is of a trans (E) structure, the double bond of the cinnamate group generates a cis (Z) structure upon irradiation with light due to a photoisomerization equilibrium. The resulting cis isomer is of a steric structure preferable for forming an intramolecular amide bond and rapidly releases an amino acid at the same time when a coumarin analogue (carbostyril) is generated.
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Killos Paul J.
Laboratory of Molecular Biophotonics
Leydig , Voit & Mayer, Ltd.
Tucker Zachary C.
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