Drug – bio-affecting and body treating compositions – In vivo diagnosis or in vivo testing – Diagnostic or test agent produces in vivo fluorescence
Reexamination Certificate
1998-11-19
2001-10-23
Hartley, Michael G. (Department: 1619)
Drug, bio-affecting and body treating compositions
In vivo diagnosis or in vivo testing
Diagnostic or test agent produces in vivo fluorescence
C424S009610, C530S363000
Reexamination Certificate
active
06306367
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to conjugates for distinguishing unhealthy or pathological tissue from healthy tissue, a process for the preparation of such conjugates as well as their use.
BACKGROUND OF THE INVENTION
When pathological tissue is treated, its removal is often an essential step. For this purpose, it is necessary for the operating surgeon to understand clearly where pathological tissue ends and healthy tissue begins. However, this is often impossible. Therefore, spurs of pathological tissue are overlooked which then represent the basis for new formation of the pathological tissue.
Thus, it is an object of the present invention to provide a method of making a distinction between pathological tissue and healthy tissue.
BRIEF DESCRIPTION OF THE INVENTION
According to the invention, this is achieved by the subject matter defined in the claims.
Therefore, the present invention relates to a conjugate comprising a fluorescence-capable compound or a compound capable of emitting fluorescence, a linker and a protein.
The expression “fluorescence-capable compound or a compound capable of emitting fluorescence” includes compounds of any kind that can be stimulated to emit fluorescence. Examples include fluorescent dyes, e.g. xanthene dyestuffs such as fluorescein, aminofluorescein (AFl), erythrosin, aminoerythrosin (AEros), eosin yellowish and 2-aminoeosin yellowish (AEs) as well as derivatives and analogues thereof. The above compounds may also have photodynamic activity. An example thereof is AEros. The photodynamic activity is typically stimulated at a wavelength ranging from 500 to 550 nm (AEros: 533 nm). Porphyrins, chlorins and bacteriochlorins are normally exempted from the above compounds.
A conjugate according to the invention may contain several compounds that are capable of emitting fluorescence and may be the same or different from one another.
A protein, such as a particularly native protein that is not considered exogenous, may be present in a conjugate according to the invention. The protein preferably has a molecular weight of up to 100,000 daltons, particularly 30,000 to 100,000 daltons. In especially preferred embodiments, the protein is albumin, particularly human serum albumin (HSA), and transferrin. It is also possible to use protein fragments. Like the proteins, protein fragments may effect a concentration of the conjugate in pathological tissues, particularly in tumor tissue and in superficial relatively small vessels, e.g. neovascularizations in the region of the cornea (keratoderma of the eye).
The term “linker” comprises compounds of any kind that are suited to link the compound capable of emitting fluorescence and the protein. Linkers are preferably not cleaved in the body. Examples of such linkers are cyanuric chloride (Cy) and derivatives thereof.
The components of a conjugate according to the invention may be given as educts. In the conjugate, they are present in derivatized form. Preferred conjugates according to the invention are shown in FIG.
1
.
Conjugates according to the invention may be prepared according to conventional processes by which the compound capable of emitting fluorescence, the linker and the protein are bonded, preferably covalently, with one another. In this connection, reference is made to the preparation of the conjugates of Examples 1 to 3 by way of example.
Conjugates according to the invention preferably have an increased half life in the organism. In addition, conjugates according to the invention preferably concentrate in pathological tissues, particularly in tumor tissue and in superficial, relatively small vessels, e.g. neovascularizations in the region of the cornea. Compounds capable of emitting fluorescence are stimulated by light, e.g. a UV lamp, so as to make visible pathological tissues, whereas healthy tissue in which the conjugates according to the invention do not concentrate are not made visible. Therefore, it is possible to delimit pathological tissues from healthy tissue.
Furthermore, the conjugates according to the invention may be stimulated to develop a photodynamic activity at a wavelength ranging from 500 to 550 nm. Light having this wavelength has only a low penetration depth in the body. Thus, the conjugates according to the invention are suited in the best possible manner to treat superficial pathological tissues, e.g. neovascularizations and esophageal tumors.
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Kaus Michael
Maier-Borst Wolfgang
Schrenk Hans-Hermann
Sinn Hansjorg
Stehle Gerd
Deutsches Krebsforschungszentrum Stiftung des offentlichen Recht
Halluin Albert P.
Hartley Michael G.
Howrey Simon Arnold & White , LLP
Kung Viola T.
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