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JP-7855565-B2 - UV-absorbing polymer dyes and methods for using them

JP7855565B2JP 7855565 B2JP7855565 B2JP 7855565B2JP-7855565-B2

Inventors

  • リャン，ヨンチャオ
  • ウッカート，フランク ピー．
  • バーソロミュー，グレン ピー．
  • レナード，バリー イー．
  • ゲイロード，ブレント，エス．

Assignees

  • ベクトン・ディキンソン・アンド・カンパニー

Dates

Publication Date
20260508
Application Date
20231214
Priority Date
20150312

Claims (13)

  1. 6-5-6 condensed tricyclic comonomers, It contains a conjugated segment that includes an ultraviolet absorbance modified comonomer, It has a maximum ultraviolet absorption capacity, Equation (I): As shown by, F1 is a 6-5-6 condensed tricyclic comonomer, b is either 1 or 2, e is either 0 or 1, a, c, d, and f are each independently either 0 or 1, where a + c + d + f ≥ 1. M1 and M2 are each independently the UV absorbance modified comonomers, When e is 0, the UV absorbance modified comonomer is the following multiple comonomers It is one of the following, n is 1 to 20, R' is an alkyl group containing 1 to 6 carbon atoms or H , R11 to R14 are each hydrogen or a water-soluble group, and at least one of R11 to R14 is a water-soluble group. When e is 1, the UV absorbance modified comonomer is R11 to R14 are each hydrogen or a water-soluble group, and at least one of R11 to R14 is a water-soluble group. L1 is a connectable comonomer containing the chemoselective tag- Z1 , The aforementioned chemoselective tag- Z1 is a functional group that can selectively react with other compatible functional groups to form a covalent bond, such as a thiol group, maleimide group, iodoacetamide group, amine group, carboxylic acid group, active ester group of a carboxylic acid, or a group that can react with each other via Click chemistry. n is an integer between 1 and 10,000. m is an integer between 0 and 10,000. p is an integer between 1 and 100,000, and G1 and G2 are each independently selected from the group consisting of terminal groups, π-conjugated segments, linkers, and linked specific-binding members, and when e is 0, G1 or G2 is a linked specific-binding member. Water-soluble, light-gathering, multi-chromophore.
  2. Formulas (II) to (IV): As shown by one of the following: The multi-chromophore described in claim 1.
  3. the below described One of the following will be selected: R 1 The following As shown by The multi-chromophore described in claim 1.
  4. L1 is structure: This is shown by, in the formula, R3 is a substituted alkyl group containing a water-soluble group. R4 is L2 - Z2 , where L2 is a linker and Z2 is a chemoselective tag. The multi-chromophore according to claim 2.
  5. A water-soluble light-gathering multi-chromophore according to any one of claims 1 to 4 , A polymer tandem dye comprising an acceptor chromophore covalently linked to the multichromophore near its energy-receiving location.
  6. The polymer tandem dye is given by formula (V): This is shown by, in the formula, F1 is a 6-5-6 condensed tricyclic comonomer, M1 and M2 are each independently ultraviolet absorbance-modified comonomers. b is either 1 or 2, a, c, d, and f are each independently either 0 or 1, where a + c + d + f ≥ 1, e is 1 , L1 is a connectable comonomer linked to the acceptor chromophore- C1 , n is an integer between 1 and 10,000. m is an integer between 1 and 10,000. p is an integer between 1 and 100,000, and G1 and G2 are each independently selected from the group consisting of terminal groups, π-conjugated segments, linkers, and linked specific-binding members. The dye according to claim 5.
  7. L1 is structure: This is shown by, in the formula, R3 is a substituent containing a water-soluble group, R4 is L2 - Z2 , where L2 is the linker and Z2 is the acceptor chromophore. The dye according to claim 6 .
  8. At least one of G1 and G2 is -L3 -Z, where L3 is a linker and Z is a specific bonding member. The dye according to claim 6 .
  9. The dye according to claim 8 , wherein Z is an antibody.
  10. The dye according to claim 8 , wherein Z is an antibody fragment .
  11. C1 is linked to L1 via an optional linker and is selected from the group consisting of cyanine pigments, xanthene pigments, coumarin pigments, thiadin pigments, and acridine pigments. The dye according to claim 6 .
  12. A method for evaluating a sample for the presence of a target sample, (a) A step of bringing a polymer dye conjugate that specifically binds to the target sample into contact with the sample, wherein the polymer dye conjugate (i) A water-soluble light-gathering multi-chromophore according to any one of claims 1 to 4 , A method comprising the steps of (ii) including a specific binding member, wherein G1 or G2 is a linked specific binding member, and (b) assaying a sample that has come into contact with the polymer dye conjugate to determine whether the target sample is present in the sample.
  13. The method according to claim 12 , wherein the target sample is associated with cells.

Description

Cross-reference of related applications In accordance with 35 U.S.C. §119(e), this application claims priority to the filing date of U.S. Provisional Patent Application No. 62/132,449, filed March 12, 2015, the disclosure of which is incorporated herein by reference. Fluorescent dyes are compounds that, when irradiated with light of a wavelength they absorb, (usually) emit light of a different wavelength. Fluorescent dyes have been found in a variety of applications in biochemistry, biology, and medicine, for example, in diagnostic kits, microscopy, or drug screening. Fluorescent dyes are characterized by many parameters that allow the user to select the appropriate dye according to the desired purpose. These parameters include the maximum excitation wavelength, maximum emission wavelength, Stokes shift, extinction coefficient, fluorescence quantum yield, and fluorescence lifetime. Dyes may be selected according to the intended application, for example, to allow the transmission of excitation radiation into a biological sample, to minimize background fluorescence, and/or to achieve a high signal-to-noise ratio. Molecular recognition requires the specific binding of two molecules. Molecules exhibiting binding specificity to target biomolecules have been found for use in various research and diagnostic applications, such as sample labeling and separation, flow cytometry, in situ hybridization, enzyme immunoassay (ELISA), Western blotting, magnetic cell separation, and chromatography. Target biomolecules can be detected by labeling with fluorescent dyes. The fluorescence emission profiles of various polymer tandem dyes based on exemplary multichromophore core structures of the disclosure of the present invention, linked to various different acceptor chromophores, are illustrated.The ultraviolet absorption spectra of various control multichromophores, MC-1 to MC-5, are shown as examples. definition Before describing exemplary embodiments in more detail, the following definitions are given to explain and define the meaning and scope of terms used herein. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art in the field to which this invention pertains. Many of the general meanings of the terms used herein are given to those skilled in the art from Singleton, et al., *Dictionary of Microbiology and Molecular Biology*, 2D ED., John Wiley and Sons, New York (1994), and Hale & Markham, *The Harper Collins Dictionary of Biology*, Harper Perennial, N. Y. (1991). Furthermore, certain terms are defined below for clarity and ease of reference. It should be noted that the singular forms “a,” “an,” and “the” as used herein and in the appended claims include the plural unless explicitly specified in the context. For example, the term “primer” means one or more primers, i.e., a single primer and multiple primers. It should be further noted that the claims may be written to exclude any element. This statement itself is intended to serve as a preceding basis for using such exclusive terms as “alone,” “only,” etc., or for using “negative” limitations in relation to the detail of the elements of the claims. As used herein, the term “sample” means a material or mixture of materials, which may be liquid, containing one or more specimens of the subject. In some embodiments, the term, in its broader sense, means plant, animal or bacterial material containing cells or producing intracellular metabolites, such as tissues or bodily fluids isolated from an individual (including, but not limited to, plasma, serum, cerebrospinal fluid, lymph, tears, saliva and tissue sections), or components of cell cultures in vitro, as well as samples from their environment. The term “sample” may also mean “biological sample.” As used herein, the term “biological sample” means a whole organism or a subset of its tissues, cells, or components (including, but not limited to, body fluids such as blood, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, amniotic fluid, amniotic umbilical cord blood, urine, vaginal fluid, and semen). “Biological sample” may also mean, but not limited to, a whole organism or a subset of its tissues, cells, or components, or a fraction or part thereof, such as plasma, serum, cerebrospinal fluid, lymph, skin, the respiratory tract, the intestinal tract, and the external sections of the urogenital tract, tears, saliva, milk, blood cells, tumors, and organs, or homogenates, lysates, or extracts prepared from such fraction or part thereof. In certain embodiments, the sample is taken from an animal or a plant. The biological sample may include cells. The term “cell” is used in its conventional sense to mean the basic structural unit of both eukaryotic and prokaryotic organisms having at least a nucleus and a cell membrane. In certain embodiments, cells include prokaryotic cells, such as cells derived from bacteria. In othe