TW200401034A - Detection of glucose in solutions also containing an alpha-hydroxy acid or a beta-diketone - Google Patents

Abstract

Compositions and methods for determining the presence or concentration of glucose in a sample which may also contain an alpha-hydroxy acid or a beta- diketone. The method uses a compound having at least two recognition elements for glucose, oriented such that the interaction between the compound and glucose is more stable than the interaction between the compound and the alpha-hydroxy acid or beta-diketone, such that the presence of the alpha-hydroxy acid or the beta-diketone does not substantially interfere with said determination.

Description

200401034 发明 Description of the invention: Reference text before and after the related application The invention was a part of the application sequence No. 10 / 〇29,1 84, which was filed on December 28, 2001, and continued on January 5, 2001. Part of the application No. 09 / 754,21 No. 7 filed in the continuation of the application, and claims that the application No. 60 / 363,885 filed on March 14, 2002, in 2001 The benefits of application preamble No. 60 / 329,746 filed on the 18th of May and application preamble No. 60 / 209,887 filed on February 21, 2001. TECHNICAL FIELD The present invention relates to the detection of glucose in samples that also contain potentially interfering compounds such as alpha-hydroxy acid or beta-one. The prior art has long known the complexation of tritium compounds (including glucose) and phenylboronic acid, and the reversibility of this parent interaction is used as the basis for carbohydrate separation. In particular, Lorand and Edwards (^ Edwards: ^ proposed the association constants of phenylboronic acid and many saturated polyols in 1959); the parent interaction system is very weak (for example, ethylene glycol, Kd = 36〇mg molecular weight) ranked to medium intensity (for example, glucose, Kd = 9. 丨 mg molecular weight). Refer to BiOrganic Medicinal Chemistry 1 (4) .267-71 (1993) by Yoon et al. The letter A binding mechanism occurs through the bonding of a few bases on glucose to a few bases on the salt moiety. U.S. Pat. No. 5,503,770 (James et al.) Describes a human sugar Fluoroboric acid-containing 85315 200401034 compound that emits high-intensity fluorescence when bound (including glucose). Fluorescent compounds have molecules containing a fluorophore, at least one phenylboronic acid moiety 6, and V to a molecule that provides a nitrogen atom. Structure, in which the nitrogen atom system is arranged near the phenylboronic acid portion, so the interaction with boric acid is used in the molecule when it is combined with sugars, and this interaction therefore causes the compound to emit fluorescence. J. Am. Chem. Soc. 117 (35): 8982-87 (1995). Also known in the art is a light-sensing agent that detects blood sugar using compounds containing onion boric acid. For example, j. Yin et al., j. Am. Chem. soc 114: 5874-5875 (1 992) shows that anthracenylboronic acid can be used as a signaling carbohydrate binding (including glucose and fructose) fluorescence photochemistry Unfortunately, compounds that interact with glucose in the manner described above also have a tendency to interact with other compounds that have hydroxyl groups, so reducing the specificity of the glucose test 'especially when the test may include interfering amounts of lactate, acetic acid, etc. 4 body samples such as salt. For example, some patients with diabetes also develop lactic acidosis 'wherein the blood lactate value is greater than 5 millimoles per liter. Therefore' it is relatively insensitive to potential interference with meridian compounds such as lactate There is still a large demand for the determination of glucose. SUMMARY OF THE INVENTION In one aspect of the invention, it relates to the determination of glucose or its concentration method in samples that also contain alfa, acid, or beta-diketones, including: a )will The product is exposed to a combination of discriminating elements with at least two glucoses 85315 200401034 and dioxin. The interaction between the compound and glucose is more stable than the interaction between the compound and alfa · ㈣ or beta · dione. The compound also contains a detectable portion with a detectable trait. When the compound is exposed to grape pupae in the sample towel, the trait changes according to the concentration method; and any change in the quantity and trait can be measured in The presence of glucose or citrate in this sample; where the presence of alpha-hydroxy acid or beta-diketone does not substantially interfere with the assay. In another opinion this month, it is about compounds with the following structure

-Ri is the same as or different from R2, and is selected from the following: 丨) hydrogen; ii) the pKa of the rs moiety and the hydrolyzable stability of the substituent, which can detect the moiety or Polymer base attached to the base, the support or matrix includes a detectable part if necessary; -I-based hydrogen or a linker capable of attaching to a solid support or polymer matrix ' · Include * detectable portion; -R4 and R5 are the same or different, and are selected from the following: i) hydrogen; ⑴ change pKa of 8 parts and hydrolyzable substituents iii) detectable portion Or 1V) a linker capable of attaching a solid support or a polymer matrix 'the support or matrix includes a detectable portion as needed;-each Z is independently carbon or nitrogen; 85315 200401034-R6 is the same as I or Not the same, and is i) a linker with 0 or 10 adjacent or branched carbon and / or heteroatoms or ii) a linker capable of attaching to a solid support or polymer matrix, the support or matrix including as needed Detectable moiety; -R is selected from the following: i) aliphatic and / or aromatic spacers, including from 1 to 10 selected from carbon, Adjacent atoms of a group consisting of nitrogen, nitrogen, sulfur, and phosphorus; η) detectable moiety or Ui) linking group capable of attaching to a solid support or polymer matrix 'The support or matrix includes detectable if necessary Parts;-each Re is the same or different, and is a part that needs to be protected, when it is not protected, it can interact with a glycol group existing near glucose; and the same or different from R10 The same, and i) hydrogen; i0 can detect a part or Hi) group, which is a) a linking group capable of attaching to a solid support or a polymer matrix, the support or matrix including a detectable part as needed (And / or dagger) include functional groups capable of modifying the physical properties of the compound; however, the book-labeled compounds include at least one detectable moiety that is associated with or directly or becomes part of a solid support or polymer. In another aspect of the present invention, the present invention relates to a detection system including the above-mentioned humanized person. σ Embodiment Person: In one aspect of the present invention, it is provided to detect glucose or its concentration in a sample, which also includes interfering chemistries. These potential interfering compounds are phosphonium butyric acid and the like. The use of acetic acid and acetic acid can distinguish glucose in a sample, but M is less likely to discriminate. 85315 200401034 A marker compound compound that interferes with the sample. The present invention. The compound has at least two identifying elements of glucose, and the interaction between the labeled compound and glucose is more stable than the interaction between the labeled compound II and the interfering compound. The general discriminant elements include those that can better reverse the crossover with glucose, especially with the glycol groups present in glucose. It is known that many discriminating elements' are preferably boric acid, borate ion, arsenite, quinite ion, telluric acid, tellurate ion, germanic acid, germanate ion, and the like. The best identification is boron. Of course, the discriminating element can be closed with a protective group, i until use. These are well known protecting groups and include neopentyl alcohol, pinacol and the like. In a specific embodiment, the blocked discrimination element is deblocked in the medium in which the compound is to be used (see, for example, Examples). The discrimination elements are preferably spaced at a suitable distance from each other on the marker compound, allowing at least two The interaction of each discriminating element with glucose results in an increase in specificity. Usually discriminative elements can have up to M atoms between each other: a spacer. It is better to orient the discriminating elements so that they can interact with glucose Separated by about 6 angstroms. The marker compound of the present invention can detect traits that change in concentration when the compound is exposed to a sample containing glucose. Many of these traits are known and can be used in the present invention. For example, marker compounds It may include a luminescent (fluorescent or fluorescent) or chemiluminescence moiety, an absorption-dominated moiety, etc. The marker compound may include an energy imparting moiety and an energy acceptor moiety, each of which makes the proposed dagger compound #Grape interaction has detectable changes. Marker compounds can include fluorophores and photo-inhibited zibu structures into 85315 -10- 200401034 Lets know the configuration of inhibiting the fluorophore with a photoinhibitor in the absence of glucose. In this case, in the presence of glucose, the configuration of the marker is changed, which causes the photoinhibitor to move to the fluorescence The group has sufficient distance, so it emits fluorescent light. On the contrary, the fluorescent group and the photoinhibitor can be structured so that they are fully isolated and emit fluorescent light with the fluorescent group in the absence of glucose; On the day of the interaction with glucose, the fluorophore and the photoinhibitor move to a close enough distance to cause photoinhibition. On January 5, 2001, it was proposed as "Detection 〇f Analytes" For the title of the joint trial with us, please refer to the preamble No. 09/7 54, 2 i 9 for a more detailed explanation of the concept of configuration changes' incorporated here for reference. Another option is that the marker can Includes parts such as fluorophores that can interact with the discriminating element or another part about the spatial configuration of 7G elements, so that the fluorophore emits fluorescence in the absence of glucose. Once added Glucose Competing glucose with the interaction between the fluorophore and the discerning element, or with the interaction between the fluorophore and other parts that are spatially arranged with respect to the discriminating element, causes the fluorescence to decrease. Example 6 illustrates this concept The real you]. A recognizes that I choose to distinguish between the fluorophore and the distinguishing element. Another part of the configuration of the element M is in the absence of grape_Tang. ^ The fluorophore does not emit fluorescence when the parent interacts. Markers that emit light or emit a relatively low fluorescence value. Once the interaction between the grape cluster and the discerning element is added, it is increased between other parts of the space configuration. When sugar is' the glucose is linked to the fluorescence or the The fluorophore competes with the parent interaction on discriminators, causing other detectable parts of the fluorescence including those whose fluorescence is affected by glucose interactions induced by electron transfer or induced effects of 85315 -11-200401034 The part. These include the U.S. Application Proceedings No.

No. 09 / 265,979 (and as published in PCT International Application WO 99/46600 on September 16, 1999) lanthanide chelates (herein incorporated by reference), polyaromatic hydrocarbons and others Derivatives, coumarin, Bompy, dansyl, catechol and the like. The other part of the category includes those whose absorption spectrum changes when the labeled compound interacts with glucose, including alizarin red. Another category includes those that regulate their fluorescence with a proximity effect. ΊΜτ7 For example, this amount is given to a donor / receptor pair, such as dansylsulfonyl / dabsyl, and the like. It is better to detect traits that can be measured, such as absorption characteristics (such as absorption rate and / or spectral drift); fluorescence disappearance time (measured by measuring time or frequency domain), glory intensity, glory Changes in anisotropy or polarization; changes in the spectral shift of the emission spectrum; changes in the disappearance of time-resolved heterogeneity (measured in the measurement time or frequency domain), etc. If necessary, the target compound of the present invention, which is supposed to have solubility, can be used directly in the solution. On the other hand, assuming that the intended application requires it, the marker compound can be immobilized (such as mechanically, or by valence or ion attachment) on an insoluble surface or a matrix, polymer, etc. When the labeling compound is trapped in, for example, another polymer, the trap should preferably penetrate into glucose sufficiently to allow a suitable interaction between glucose and the labeling compound. If the marker compound is not very soluble or insoluble in water, it is not desirable to copolymerize the marker compound with a hydrophilic monomer in the medium to form 85315 • 12 · 200401034. The hydrophilicity of the compound is large, such as in 2_year M 4 Japan has filed a joint trial of US Application No. _32,624, which is incorporated herein for reference. ,

I Preferred label compounds have the following structure:

Where -He is the same or different from R2 'and is selected from the following: i) hydrogen; ii) a substituent that changes the pKa and hydrolytic stability of the R8 moiety, Π) can detect the moiety or can be compatible with the solid-side support or Polymer base attached to the linker, the support or matrix includes a detectable part as needed;-R3 series hydrogen or a linker capable of attaching to a solid support or polymer matrix, the remote support or matrix includes彳 Measured part;-R4 and Rs are the same or different, and are selected from the following: 丨) hydrogen; π) the pKa and hydrolyzable substituent-, iH) can be detected part or iv) a linking group capable of attaching to a solid support or polymer base, the support or base

I includes detectable parts if necessary;-each Z is independently carbon or nitrogen;-R_6 is the same as or different from R? And i) has 0 or 10 adjacent or branched carbons and / Or a heteroatom linker or ii) a linker capable of attaching to a solid support or polymer matrix, the support or matrix including a detectable moiety as needed; -R is selected from the following: i) aliphatic and / Or aromatic spacer, which includes from 1 to 85315-13 · 200401034 10 adjacent atoms selected from the group consisting of carbon, oxygen, nitrogen, sulfur and phosphorus; ii) a detectable moiety or iii) capable of interacting with A solid support or a linker attached to a polymer matrix 'The support or matrix includes a removable portion as needed;-each Rs is the same or different and is a portion that needs to be protected when it is not protected 'It can interact with glycol groups existing near glucose; and -9 and 111 () are the same or different and are i) hydrogen; ii) detectable part or ni) groups' it is a ) A linker capable of attaching to a solid support or polymer matrix; the support or matrix includes a measurable portion as needed, and / or b) includes Change the physical properties of the compound having a functional group; the proviso based compound comprises at least one marker or directly entered thereto into a solid carrier or a polymer or a part of the detectable portions. Those who are generally familiar with this technology can easily understand the suitable pKa and hydrolytically stable bases for modification & part, and include halogen'nitro, amine, halogen-substituted alkyl, and optionally substituted carboxyl, Amidino, keto, nitrile, amidine, vinegar, alkoxy and the like. Suitable linkers for any substituent may include from about 1 to about 20 contiguous atoms, which may be branched or substituted, and may include one or more heteroatoms, which can be further reacted or reacted with a polymer or The functional group attached to the support is terminated. Examples of suitable linking groups include alkyl, aryl, fluorenyl, polyamide, polyphenol, all such groups and combinations thereof optionally substituted. And can further include functional groups that can change the physical properties of the compound, such as the degree of resolution, PKa and so on. These include, for example, carboxylic acid esters, amine groups, quaternary ammonium groups, sulfonate esters, PEG, and the like, which are optionally substituted. 85315 -14 · 200401034 Of course, when any substituent is a detectable moiety, it may also include a suitable linker 'linking the detectable moiety with the rest of the marker compound. Suitable linking groups include those listed above. Suitable detectable parts include those defined above. R8 is preferably selected from the group consisting of boric acid, pentoxide ion, quinconic acid, quinuclide ion, telluric acid, tellurate ion, germanic acid, germanate ion, and combinations thereof. It is also understood from the above definition that the compounds and detection systems of the present invention may have a polymer form. Therefore, the entire compound (including the identifying element and the detectable moiety) can be linked to the existing polymer, or the entire compound in the form of a monomer can be polymerized or copolymerized with another suitable monomer to form a polymer. Another option is to copolymerize two separate monomer components (one including a discriminating element and one including a detectable moiety), so the resulting polymer includes all the elements necessary for the system (see Example 6). The marker compound of the present invention has many uses, including use as a marker in the fields of energy, medicine, and agriculture. For example, marker compounds can be detected in physiological buffers or physiological fluids (such as blood, plasma, serum, interstitial fluid, cerebrospinal fluid, urine, saliva, crystalline fluid, 'lymph, tears, or sweat). Low or too high glucose values, thus providing valuable information for the diagnosis or monitoring of diseases such as diabetes or adrenal insufficiency. There is a need to monitor and control the medical / agricultural production of glucose for human therapeutic applications. The use of the present invention in agriculture includes detecting glucose values in soybeans and other agricultural products 85315-15 · 200401034. Care must be taken to monitor the glucose of these high-value products, such as wine grapes, at critical harvest decisions. When glucose is the most expensive carbon source and raw material in the fermentation process, it is important to monitor the optimal S: glucose reactor feed rate control when there is authoritative wine production. Reactor mixing and glucose concentration control are also key to quality control during beverages and fermented beverages, which consume large amounts of glucose and internationally fermentable sugars (nearly glycols). When a label compound is incorporated into a fluorescent label substituent, various detection techniques are also known in the art. For example, the compounds of the present invention (e.g., U.S. Patent No. 5,517,313) can be used in a fluorescence sensing device or can be combined with polymers, such as test papers for visual inspection. This latter technique may allow glucose to be measured, for example, in a manner similar to determining pH with litmus paper. The compounds described herein can also be used as simple reagents for standard desktop analytical instruments, such as Shimadzu, mtachi,

Fluorometers or clinical analyzers manufactured by Jasco, Beckman, and others. These molecules can also provide optical fiber-based analyte-specific chemistry based on Ocean Optics (Dunedin, FL) or Oriel Optics M / Optical signalling. U.S. Patent No. 5,5 17,313 (the disclosure is incorporated herein by reference) describes a fluorescence sensing device in which a compound of the present invention is used to determine the presence of glucose or its degree in a liquid medium. The sensing device includes a layered array of a matrix containing fluorescently labeled molecules (hereinafter referred to as a "fluorescent matrix"), a high-pass filter, and a light detection device. In the Rong device, the light source (preferably a light-emitting diode ("LED")) is at least partially located in the substrate or in the waveguide. When a marker 85315 -16 · substrate is arranged, the light source The incident light causes the marker molecules to emit light. The high-pass filter allows the emitted light to reach the photodetector, while filtering the scattered incident light from the light source. Regulated (eg, attenuated or enhanced) glucose in the presence of a marker molecule used in the device described in U.S. Patent No. 5,517,313. In the sensor described in U.S. Patent No. 5,5,17,3,13, a substance containing a marker molecule can pass through the analyte. Therefore, the 'analyte can diffuse into the substance from the periphery of the test medium, thereby affecting the light emitted by the labeled compound. The structure composed of a light source, a substance containing a marker compound, a Nantong wave device, and a light sensor makes at least part of the light-emitting collision light detector 'producing the electric 彳 s' emitted by the marker compound, which is connected to Indication of glucose concentration around the medium. According to other possible embodiments of using the marker compound of the present invention, it is also described in U.S. Patent Nos. 5,9,0,661, 5,9n, 605, and 5,894,351. The description of the sensing device is incorporated herein by reference in its entirety. The compounds of the present invention can also be used in implantable devices that continuously monitor blood glucose levels in vivo, for example. It is described in, for example, U.S. Application Serial Nos. 09 / 83,148, filed on August 26, 1999 and U.S. Patent Nos. 5,833,603, 6,002,954, and 6,0U, 984. Suitable devices are incorporated herein by reference in their entirety. The compounds of the present invention can be prepared by those skilled in the art using readily available reaction mechanisms and reagents (e.g., including reaction mechanisms that conform to the general procedures described below) without undue experimental amounts. f Example 1 85315 -17- 200401034 Water-soluble copolymer of anthracene derivative and MAPTAC I. Synthesis of mono-borate-anthracene label copolymerized in water-soluble polymer: A. 9- [3- (methyl Propidylamino) propylamino] methylanthracene DIE A (1 8.5 g '25.0 ml, 144 mmol, 6.5 equivalents) was added dropwise over 20 minutes to N- in 250 ml of CHC13 at 0 ° C (3-Aminopropyl) methacrylamidine hydrochloride (11.82 g, 66.0 mmol, 3.0 eq) and DBMP (10 mg as a marker) in suspension. The mixture was allowed to warm to 25 ° C and then cooled to zero. 〇. A solution of 9-chloromethyl mercapto (5.0 g) '2 2 Emole in CHC13 (100 ml) was added dropwise to the cooled mixture over 1 hour. The mixture was then stirred at 25 ° C for 1 hour, then at 5 ° C for 12 hours and then at 7 ° C for 2 hours. At this point the mixture was washed with 4 x 60 ml portions of water and the combined aqueous layers were Extracted with CH2C12. The combined organic extracts were dried over anhydrous NasSCU, decanted and concentrated in vacuo. The crude material was purified by silica gel layer separation (flash silica, 2_5 ° / 〇CH3OH / CH2Cl2) to produce 2.44 G (33%) solid product. TLC: Merck Silicone 60 flat plate, Rf 0.39 Observed with 90/10 CH2Cl2 / CH3OH, UV (254/366), indan trione stain. Β · 9- [N- [2- (5,5-dimethylborofluoren-2-yl) benzyl] -N- [3- (methylpropionamido) propylamino] methylanthracene Grams of '3.84 ml, 22.0 mmol, 3.0 equivalents) 9- [3- (fluorenylpropenamido) propylamino] methylanthracene (200-methylpropenylamido) propylamine] in 200 ml of CHCb in portions at 10 ° C over 10 minutes 2.44 grams, 7.34 millimoles) and DBMP (10 milligrams as a marker), followed by dropwise 85315 -18-(2-bromomethylphenyl) neopentyl borate (2.49 G, 8.8 1 Moore, 1.2 equivalent) solution. The mixture was then stirred at 25. (: for 20 hours. At this time the mixture was washed with water 'and the combined aqueous layers were extracted with CH2C12. The combined organic extracts were dried over anhydrous NajO4 , Decanted and concentrated in vacuo. The crude material was purified by JS gelatin layer separation method (flashed stone gelatine, 2-5% CH3OH / CH2Cl2), yielding 2.50 grams (76%) of light yellow A crystalline solid.

Melting point: 72-73 ° C TLC: Mok silica gel 60 flat plate, Rf 0.36 was observed with 90/10 CH2Cl2 / CH3OH, U V (2 5 4/3 6 6) 'section was filled with Mikasa stain. C. 9- [N- [2- (5,5-dimethylborofluoren-2-yl) benzyl] -N- [3- (methylpropylamido) propylamino] methylanthracene and The water-soluble copolymer of MAPTAC (1:20 mole ratio) will be 4,4'-azobis (cyanovaleric acid) (0.008 g, 0.03 millimoles, 14 mole%). Total monomers) 9- [N- [2- (5,5-dimethylboran-2-yl) fluorenyl] -N- [3- (methylpropylpropane) in 1.5 ml of ethylene glycol Hydrazine) propylamino] methylanthracene (0.0490 g, 0.105 mmol) and [3- (methylpropylalanino) propyl] trimethylammonium chloride (MAPTAC '50 wt./0 in water 〇48g, 0.90 **: liter, 2.1 millimoles, 20 equivalents) in solution. The solution was flushed with argon for 5 minutes and then heated to 60 ° C in the dark for 18 hours. At this point the viscous solution was cooled to 25 ° C, diluted with 5 ml of water and dialyzed through a cellulose acetate film (MWCO 3500) with 3x4 liters of water. The dialysate was concentrated to dryness, yielding 0.339 g (68%) of a yellow glassy solid. Π. Fluorescence-modulating effect with glucose and lactate The copolymer (which includes a single discriminating element) prepared in this example was measured with the 85315 • 19-glucose and lactate fluorescence-modulating effect β chart! Shown in a normalized fluorescent solution comprising a) 0-20 mg molecular weight glucose; b) 0-20 mg molecular weight lactate in pBS = gram / ml copolymer solution (molar ratio of 1:20) Light hair $ (1 / 1〇 @ 420nm). A Shimadzu RF_53〇1 fluorometer was used to record the spectrum ', which was excited with @ 365 nm, an excited aperture of u nm, an emission aperture of 5 μm, and room temperature. Error bars are the standard deviation of the repeated values for each data point. The fluorescence of the copolymer is affected by the presence of glucose and acetate. Example 2 Covalently attached to a water-soluble polymer ¥ Bi · boronium salt_ Regulatory effect of glucose on glucose and potential physical interference I · Bi · borate-anthracene-labeled single methylacrylate monomer Synthetic effect

Add 9,10-bis (chloromethyl) anthracene (3.94 g, 14.3 mmol) to 2- (2-aminoethoxy) ethanol (3 1.4 g, at 40 ° C in CHCI3 at 23 ° C) 30.0¾ liters' solution in 299 millimoles, 20.9 equivalents). The solution was stirred in the dark for 67 hours. At this time, 100 ml of CH2C12 was added, and washed with 1 x 50 ml and 2 x 100 ml portions of NaHC03 (a saturated aqueous solution). The organic extract was dried over anhydrous Na2SO4, filtered and concentrated to give 4.67 g (79%) of a yellow powder. The reaction of 85315-20.200401034 was continued as it was (~ 85% purity by RP-HPLC). HPLC conditions: HP 1100 HPLC chromatography, Vydac 201TP 10x250 mm tube, 0.100 ml injection, 2 ml / min, detection at 370 nm, A = water (0.1% HFBA) and B ^ MeCN (0.1% HFBA ), Gradient: 10 minutes B for 2 minutes, 10-80% B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; residence time 15.6 minutes. ’

B. 9,10-bis [N- [2- (5,5-dimethylboramidine-2-yl) fluorenyl] -N- [2- (2-hydroxyethoxy) ethylamino] methyl Phenyl] anthracene, 9,10-bis [[2- (2-hydroxyethoxy) ethylamino] methyl] anthracene (4.02 g, 9.75 mmol) in -125 ml of CHC13, -DIEA (12.6 G, 17.0 ml, 97.5 millimoles, 10.0 equivalents) and (2-bromomethylphenyl) neopentyl borate (13.7 grams, 48 millimoles, 4.9 equivalents) at 23 ° C Stir in the dark for 46 hours. At this point the reaction mixture was first concentrated by rotary evaporation and then concentrated under vacuum to remove DIEA. The residue was purified by alumina 'column chromatography (150 grams of activated neutral alumina, 0-3% CH3OH / CH2C12), yielding 5.67 grams (70%) of a viscous oil, which solidified upon standing. The reaction was continued as it was (~ 85% purity by RP-HPLC). 85315 -21-200401034 TLC: Mok's basic alumina plate, Rf 0.33 is observed at 95/5 of CH2C12 / CH30H, UV (254/366). HPLC conditions: HP 1100 HPLC chromatogram, Vydac 201TP 10x250 mm column, 0.100 ml injection, 2 ml / min, detection at 370 nm 'A = water (0.1% HFBA) and B = MeCN ( 0.1% HFBA), gradient: 100 / (B) for 2 minutes, 10-80% B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; residence time 18.8 minutes.

C · 9- [N- [2- (5,5-dimethylborofluoren-2-yl) benzyl] -N- [2- (2-methylpropanyloxyethoxy) ethylamino ] Methyl-10- [N- [2- (5,5-dimethylshed-2-yl) benzyl] -N- [2- (2-hydroxyethoxy) ethylamino] methyl 1 anthracene (single fluorenyl acrylate monomer) will be in 15 ml (: 112 (: 9,10-bis. | 12 in 12) | ^-[2- (5,5-dimethylborofluoren-2-yl) Benzyl] -N- [2- (2-hydroxyethoxy) ethylamino] methyl] anthracene (0.298 g, Q.359 mmol), fluorenylpropanoic acid (0.304 g, 0.300 ml, 3.53 Millimoles, 9.84 equivalents), (: (: (0.965 grams, 4.68 millimoles, 13.0 equivalents), and N, N-dimethylaminopyridine (0.020 grams, 0.16 millimoles, 0.46 The solution was stirred at 23 ° C in the dark for 4 hours. At this time, the reaction mixture 85315 -22- 200401034 was filtered and concentrated by rotary evaporation. The residue was separated by alumina column chromatography (50 g activation) Neutral alumina, 0-4% CH3OH / CH2C12), yielded 0.150 g (47%) of a yellow solid. FAB MS: calculated C52H66B2N209 [M] + 855; experimental value [M + l] + 886. TLC: Mo Gram alkaline alumina plate Rf 0.45 with 95/5 CH2Cl2 / CH3OH and UV (254/366) observation. HPLC: HP 1100 HPLC chromatogram, Vydac 201TP 10x250 mm column, 0.100 ml injection, 2 ml / min, 370 milliliter Micron detection, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA), gradient: 10% B over 2 minutes, 10-80% B over 18 minutes, 80-100% B over 2 minutes, 100 % B after 2 minutes; residence time 21 minutes. D · 9- [N- [2- (5,5-Dimethylborofluoren-2-yl) fluorenyl] -N- [2- (2-form Propylpropyloxyethoxy) ethylamino] fluorenyl-10- [N- [2- (5,5-dimethylborofluoren-2-yl) benzyl] -N- [2- (2 -Hydroxyethoxy) ethylamino] methyl] anthracene and water soluble copolymer of TMAMA (Molar ratio of 1:50) will be in 3.00 ml] 9-[^ [-[2- (5,5-dimethylborofluoren-2-yl) benzyl] -N- [2- (2-methylpropionyloxyethoxy) ethylamino] methyl-10- [N- [ 2- (5,5-dimethylborofluoren-2-yl) benzyl] -N- [2- (2-hydroxyethoxy) ethylamino] methyl] anthracene (0.0024 g, 0.0033 mmol ) Solution was added to [2- (methylpropanyloxy) ethyl] trimethylammonium chloride (TMAMA, 70% by weight aqueous solution, 0.344 g monomer) in 0.600 ml of water 1.66 mmol, 50 eq) of a solution. 4,4'-Azobis (4-cyanovaleric acid) (0.0075 g, 0.027 mmol, 1.6 mole% total monomer) was added to the mixture. The solution was filtered through 85315-23 · 200401034 0,45 micron membrane filter paper, rinsed with 0, lice and then heated at 55 C in the dark for 16 hours. At this time. # J ， 谷 夜 冷 部To 25 t and concentrated in the air. The residue was diluted with 20 ml of water 〆, _ φ. 睪 and 二, filtered through a 0.2 micron membrane filter paper. The solution of the 4 compound was passed through the fiber γ spit fruit vinegar salt The film (MWCO 3500) was dialyzed with 2x4 liters of water. 38.5 渗 liters of polymer solution was obtained using dialysis. Concentrated part of the solution was dried to show that the mother 1.0¾ liter solution had 0.00 75 gram poly: The total yield of polymer was 0.289 g (77%). II. The copolymers prepared in this example (which includes two discriminating elements) were determined by the fluorescence regulation of glucose, lactate and acetamidine acetate. Glucose and lactic acid are used to regulate the fluorescence of acetamidine acetate. Figure 2 shows the results including a) 0-20 mg molecular weight glucose; b) 0_2 mg molecular weight lactate ·, e) 0-20 mg molecular weight B 5 5 mg / ml of anthracene diboronic acid. 犟 -TjMAMA solution in pBS of lithium acetate ( 1:50 (Molar ratio) of regularized fluorescence emission (1 / 1〇 @ 42,8nm). The spectrum was recorded using a Shimadzu RF_53〇1 fluorometer, which was excited at @ 365 驾 microns' ι · 5 nm excitation. Dance pores, 1. f μm emission pores, room temperature. The fluorescence of the copolymer is affected by the presence of glucose 'but not by the presence of lactate or acetoacetate. Example 3 In Dose-response effect of lactate on glucose in solution in bis_borate_anthracene

85315 24- 200401034 A. 9,10-Bis [[2- (tert-butoxycarbonyl) ethylamino] methyl] anthracene will be raised in 75 ml of CHC13-tert-alanine hydrobutyrate (3.06 Grams '16 .8 millimoles, 5.09 equivalents), £ 8 (4.27 grams, 5.75 milliliters, 33.0 millimoles, 10.00 equivalents), and 9,10_bis (chloromethyl) anthracene (0.910 grams, 3.3 1 milligrams) Mol) solution was stirred at 23 ° C in the dark for 93 hours. At this point, the solution was filtered and washed with U40 ml and 2 x 60 ml portions of NaHC03 (saturated aqueous solution). The organic extract was dried over anhydrous Na2S04, filtered and concentrated 'to give a crude yellow solid. The residue was purified by silica gel column chromatography (30 g gravity-grade gel, 0-3% CH3OH / CH2C12) to yield 1.06 shek (63 ° / °) sticky yellow-orange product. The reaction was continued as it was. TLC ·· Mokshi Xijiao 60 flat plate, Rf 0.33 Observed with 95/5 of CH2C12 / CH30H and UV (254/366). ·

B. 9,10-bis [N- [2- (5,5-dimethylboramidine-2-yl) benzyl] · ν_ [2_ (tert-butoxycarbonyl) ethylamino] methyl] anthracene ./ Will be 9,10 'bis [[2- (tert-butoxycarbonyl) ethylamino] fluorenyl] Zine (1.60 g, 3.25 mmol) in 30 ml CHCU, DIEA (4.45 G, 600 ml of 34.4 millimoles, 10.6 equivalents) and (2-bromomethylphenyl) neopentyl borate (4.80 grams '17 .0¾ mole '5.22 equivalents) of the solution at 23. Stir in the dark for 4′5 days. At this time, 45 ml of CHCI3 was added to the mixture, and the 85315 -25- 200401034 mixture was washed with 2 x 25 ml portions of NaHC03 (a saturated aqueous solution). The organic extract was dried over anhydrous Na2S04, filtered and concentrated to give a crude red oil. The residue was purified by oxidation column chromatography (100 grams of activated neutral alumina, 0-3% CH30H / CH2C12) to yield ~ 3.5 grams of orange solid. The product was dissolved and a white precipitate (DIEA-HBr salt) formed. The solution was filtered and the filtrate was concentrated to give 2.72 | g (93%) of an orange solid. The reaction was continued as it was (with RP-HPLC > 80% purity). TLC: Mok's basic alumina plate, Rf 0.66 observed at 95/5 CH2Cl2 / CH3OH, UV (254/366). HPLC conditions: HP 1100 HPLC chromatography, Vydac 201TP 10x250 mm column, 0.100 ml injection, 2 ml / min, detection at 370 nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA ), Gradient: 10 ° / 〇B for 2 minutes, 10-80% B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; residence time 23.9 minutes.

C. 9,10-bis [N- (2-Dihydroxyborylbenzyl) -N- [2- (carboxyethyl) amino] fluorenyl] E will be 9 in 5 ml of 20% TFA / CH2C12 , 10-bis [N- [2- (5,5-dimethylglut-2-yl) benzyl] -N- [2- (tert-butoxycarbonyl) ethylamino] methyl] anthracene (〇 .556 g, 0: 620 millimoles) was stirred at '23 ° C in the dark for 25 hours. At this point the reaction mixture was concentrated under a stream of N2. The residue was triturated with 3 x 10 milliliter 85315 -26- 200401034 liters of ether. The residual solid was dried in vacuo to give 0.35 g (87%) of a fluffy yellow powder. FAB MS: glycerol matrix; calculated C42H46B2N2ioi (bisglycerol adduct) [M] + 760; experimental value [m] + 760. HPLC: HP 1100 HPLC chromatography, Waters 5x100 mm NovaPak HR C18 column, 0.025 ml injection, 0.75 ml / min, 1.5¾ liter injection ring, 360 nm detection, water (0.10 / 〇HFBA) & B = MeCN (0, 1% HFBA), gradient: 10% B over 2 minutes, 10-80% B over 18 minutes, 80-100% b over 2 minutes, 100o / 〇B over 2 minutes; stay Time is 16.7 minutes. D. Fluorescence-regulating effect with glucose and lactate The fluorescence-regulating effect of the marker compound (which includes two discriminating elements) prepared in this example with glucose and lactate was measured. Figure 3 is shown in a pbs including a) 0-10 mg molecular weight glucose '0 mg molecular weight lactate; b) 0 10 mg molecular weight glucose, 2 mg molecular weight lactate; c) 0-10 mg molecular weight glucose' 5¾ g molecular weight lactate Of 0.75 microgram molecular weight biscarboxylate bis-rotate-anthracene labeling solution (at 428 nm). The spectrum was recorded using a Shimadzu RF-5301 fluorometer, which was excited with @ 365 nm, 1.5 μm excited pores, and $. Nanometer emission pores, chamber. All points were measured three times, including ± 1 SD error bars. The presence of lactate does not substantially affect the fluorescent regulation of the marker with glucose. Example 4 When it is known that hexamethylene is covalently fixed in a hydrogel, the selectivity of bis-borate glucose to γ-C to glucose relative to lactate and acetamyl acetate 85315 -27- 200401034 I. Preparation of di-methacrylamide monomer:

A. 9,10-Bis [3- (methylpropanilamine) propylamino] methylanthracene will be 9,10-bis (chloromethyl) anthracene (1.5 g, 5.45 mmol) in 200 ml of CHC13 Ear), DIEA (28.17 g, 38,00 ml, 218 millimoles, 40 equivalents), N- (3-aminopropyl) methylpropionamine hydrochloride (9.76 g, 54.5! Moore, 10.0 equivalents) and a suspension of ~ 5 mg BHT in agitation at 23 ° C in the dark at 40 ° C for 4 days. The temperature was increased to 45 ° C at this time, and the mixture was stirred for more than 3 days. A precipitate formed at this time. The mixture was filtered and the solid was dissolved in a minimal amount of CH2C12. A yellow crystalline solid formed overnight as a dihydrochloride salt of the expected product (3.15 g, quantitative). TLC: Mok alkaline alumina flat plate, Rf 0.31 observed at 90/10 CH2C12 / CH30H, UV (254/366). HPLC: HP 1100 HPLC chromatography, Waters 5x100 mm NovaPak HR C18 column, 0.L00 ml injection, 0.75 ml / min, detection at 3 60 nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA), gradient ': 10% B for 2 minutes, 10-80% B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; residence time 15.0 minutes. 85315 • 28-! 200401034

B. 9,10-Bis [N- [2- (5,5-dimethylboramidine-2-yl) fluorenyl] -N- [3- (methylpropylamido) propylamino] methyl Anthracene (di-methylpropylammonium monomer) 9,10-bis [3- (methylpropylamido) propylamino] methylanthracene (0.6650 g) in 20 ml of CTHC13 , 1.34 millimoles of free amines), 01 ugly (0.612 grams, 0.825 ml, 4.74 millimoles, 3.55 equivalents), (2-bromofluorenylphenyl) boric acid neopentyl ester (1.34 grams, 4.74 millimoles Ear, 3.55 eq.) And BHT (5 mg, as marker) were stirred at 23 ° C for 5 days in the dark. At this point the reaction mixture was concentrated in vacuo and the residue was purified by oxidative color separation (200 g of activated neutral alumina, 0-2% CH3OH / CH2C12), yielding 0.465 g (39%) of a very viscous Yellow oil. TLC: Mok alkaline alumina plate, Rf 0.59 was observed at 90/10 CH2Cl2 / CH3OH, UV (254/3) 6). HPLC: HP 1100 HPLC chromatography, Waters 5x100 mm NovaPak HR C18 column, 0.050 ml injection, 0.75 ml / min, detection at 360 nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA), gradient: 10% B over 2 minutes, 10-80% B over 18 minutes, 80-100% 8 over 2 minutes, 100% 6 over 2 minutes; residence time 16.9 minutes 85315 -29 minutes. C. Preparation of N, N-dimethylacrylamide hydrogel with glucose marker N, N-dimethylacrylamide (40% by weight) and N, N in ethylene glycol A solution of '-methylenebisacrylamide (0.8% by weight). The 9,10-bis [N- [2- (5,5-dimethylboron, sam_2_yl) fluorenyl] _n_ [3_ (methylpropylfluorenylamino) propylamino] 17.8 mg '2xl0.5 mole) and 40 microliters of aqueous ammonium persulfate (5 weight 0 /.) Were combined with 1 ml of ethylene glycol monomer solution. The resulting solution was placed in a glove box flushed with nitrogen. N, N, N ,, N, -tetramethylethylenediamine aqueous solution (80 liters and 5 weights) was added to the early-body formulation to accelerate the polymerization. The resulting formulation was poured into a microscope sheet and 10%. 〇micron stainless steel spacer plate structure model. After maintaining for 8 hours under nitrogen, put the model in phosphate buffered saline (PBS) (100 mg molecular weight PBS, pH = 7.4), and separate the microscope slide And remove the hydrogel. The hydrogel was washed with 100 ml of PBS including 1 mg of sodium lauryl sulfate and 1 mg of EDTA sodium salt for 3 days, and the solution was changed daily, followed by DMF / PBS (10 / volume by volume) 90, 3x100 ml) and finally washed with? 88 (mouth 11 = 7.4, 3 \ 100 ml). The resulting hydrogel polymer was stored in PBS containing 0.2% by weight of sodium azide and 1 mg of molecular weight EDTA sodium salt Medium (10 mg molecular weight PBS, pH = 7.4). II. Fluorescence modulation of glucose, lactate and acetamidine acetate was measured. The marker compound (which includes two discriminating elements) prepared in this example was glucose , Lactate and acetic acid acetate Figure 4 shows the 10 mg molecular weight 85315-30 · 200401034 PBS (pH 7.4) including 0.2% Na% and 1 mg molecular weight EDTA (including various sodium l-lactate, lithium ethyl acetate or 0: -D-glucose amount). Included in this example is the normalized fluorescence emission of a hydrogel of glucose discriminator (1 / 1〇 @ 427nm). Data was recorded using a Shimadzu RF-5301 goniophotometer, which was excited with @ 365nm ( Porosity = 3 nm) and emission at 427 nm (porosity = 3 nm), low selectivity, 'using temperature-controlled sample holder at 37 ° C. 3 ml expected The solution tube reached equilibrium at 37 ° C over 15 minutes. Each hydrogel sample was measured with 4 independent samples. The error bars are the standard deviations of the four replicates of each data point. As previously described Preparation of a hydrogel including glucose discriminators. The hydrogel was loaded on a glass sheet in a PMMA tank tube at 45 ° incident light and covered with a polyester mesh. At 10 including 0.2% NaN3 and 1 mg molecular weight EDTA Prepare 1, 5, 10, and 20 mg MW in PBS (pH 7.4) Gram molecular weight L-sodium lactate solution [Aldrich]; 5, 10 and 20 mg molecular weight acetoacetic acid noodle solution [Alderrich] and 1, 2, 4, 5, 10 and 20 mg molecular weight α -D-glucose solution. The fluorescence of the copolymer is affected by the presence of glucose, but is not affected by the presence of lactate or acetamidine acetate. Example 5 The use of bis-borate to discriminate and nearby suppressive signals produces glucose against lactic acid Selectivity of salt A. N- (2,2-diethoxyethyl) -4 -molyl-1,8-fluorenyldimethylamine 4-bromo-1,8 in 45 ml EtOH -Suspension of acetic anhydride (10.0 g, 36.1 mol) and aminoacetaldehyde diethyl acetal (4.81 g, 5.26 ml, 36.1 mmol, 1 equivalent) at 45 ° C Stir for 3 days. The resulting suspension was filtered at this time, washed with EtOH and the residue was dried to give 31.3 g 85315 -31-200401034 (94%) as a light brown solid product. TLC: Mok Silicone 60 plate, Rf 0.17 with 98/2 CH2C12 / CH30H and UV (254/366) observation. HPLC ·· HP 1100 HPLC chromatography, Waters 5x100 mm NovaPak HR C18 column, 0.050 ml injection, 0.75 ml / min, 1.5 ml injection ring, 360 nm detection, A = water (0.1% HFBA ) And β = MeCN (0.1% HFBA), gradient: 10% B over 2 minutes, 10-80% B over 18 minutes, 80-100% 8 over 2 minutes, 100 ° / (^ over 2 minutes; stay time 24.2 Min. Beta-N- (2,2-diethoxyethyl) -4-butylamino-1,8-fluorenediamine will be N- (2,2-diethyl) in 8 ml NMP (Oxyethyl) -4-bromo-1,8-fluorenedimethylamine (0.797 g, 2.03 mmol) and n-butylamine (1.48 g, 2.00 ml, 20.2 mmol, 9.96 equiv) Stir for 66 hours at 45 ° C. At this time allow the resulting suspension to cool to 25 ° C and then filter. Dissolve the residue in 50 ml of ether and extract with 3x50 ml of water. Dry the organic extract over anhydrous Na2S04, Filtration and concentration yielded a crude yellow powder. The crude material was purified by silica gel chromatography (25 grams of gravity-grade gel, 0-ΐ% CH3OH / CH2Cl2) to yield 0.639 grams (82%) of a yellow powder. TLC: Mok Silicon 60 plate, Rf 0.71 with 95/5 CH2C12 / CH30H and UV (254/366) observation. HPLC: HP 1100 HPLC chromatogram, Waters 5x100 mm NovaPak HR C18 column, 0.050 ml injection, 0.75 ml / min , 1.5 ml injection ring, detected at 450 nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA), gradient: 10% B for 2 minutes, 10-80% B for 18 85315 -32 -200401034 minutes, 80-100% 6 after 2 minutes, 100% 8 after 2 minutes; residence time 23.5 minutes. C. N- (2-oxoethyl) -4-butylamino-1,8-naphthalenedimethyl Ammonium will be N- (2,2-diethoxyethyl) -4-butylamino-1,8-dimethylformamide (0.622 g, 1.62 mmol) in 25 ml of acetone. -A solution of toluenesulfonic acid monohydrate (0.010 g, 0.053 mmol, 0.032 eq) was stirred at 25 ° C for 18 hours. At this time, the solution was filtered and the residue was separated by a silica gel layer (25 g Gravity-grade gel, 0-1% CH3OH / CH2C12) purified to yield 0.470 g (94%) of orange solid. TLC: Mok Silicone 60 plate, Rf 0.61 to 95/5 of CH2Cl2 / CH3OH, UV (254 / 366) Observe. 'H NMR (400MHz, CDC13); 5 1-〇3 (t, 3H, J = 7.3Hz), 1.53 (m, 2H), 1.78 (m, 2H), 3.38 (t, 2H, J = 7.2Hz) , 5.02 (s, 2H), 6.64 (d, 1H, J = 8.6Hz), 7.52 (dd, 1H, J = 7.4, 8.3Hz), 8.08 (dd, 1H, J = lHz, 8.5Hz), 8.38 ( d, 1H, J = 8.3Hz), 8.46 (dd, 1H, J = 1.0, 7.3Hz), 9.75 (s, 1H). HPLC: HP 1100 HPLC chromatogram, Waters 5x 100 mm NovaPak HR C18 column, 0.050 ml injection, 0.75 ml / min, 1.5 ml injection ring, detection at 450 nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA), gradient: 10% B for 2 minutes, 10-80% B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; residence time 19.6 minutes. D. 4-Difluorenylaminobenzaldehyde (1.00 g, 85315 -33-N- (4-dimethylamino) -1,6-diaminohexanoic acid in 20 ml of anhydrous EtOH 200401034 6.70 millimoles), Na2S04 (6_70 grams, 47.2 millimoles, 7.04 equivalents) and 1,6-diaminohexane (3.89 grams, 33.5 millimoles, 5.00 equivalents) at 25 ° C and Stir in the dark for 18 hours under nitrogen. The solution was filtered at this time, and NaBH4 (1.73 g'45.8 mmol, 6.84 equivalents) was added to the filter. The suspension was stirred at 25 ° C for 5 hours. At this point the reaction mixture was concentrated and the residue was dissolved in 50 ml of water and extracted with 3 x 50 ml of ether. The combined organic extracts were washed with 2x50 ml of water. The combined aqueous extracts were extracted with 2 x 50 ml of ether. The combined organic extracts were dried over NaJO4, filtered and concentrated to give 1.35 g (81%) of a viscous oil. TLC: Mok silicone 60 plate, Rf 0.58 with CH2Cl2 / CH3OH / iPrNH2 of 80/15/5, observed with indanetrione stain, v (254/366). HPLC: HP 1100 HPLC chromatography, Waters 5x100 mm NovaPak HR C18 column, 0.050 ml injection, 0.75 ml / min, 1.5 ml injection ring, detection at 280 nm, A = water (0.1% HFBA) and B =

MeCN (0.1% HFBA), gradient: 10% B for 2 minutes, 10-80% B for 18 minutes' 80-100% B for 2 minutes, 100% B for 2 minutes; residence time 13.3 minutes. E. N-2- [6-N- (N-4-Dimethylaminobenzyl) aminehexyl] aminoethyl) _oxobutylamino-1,8-naphthalenediamine will be in 20 ml N- (4-dimethylaminobenzyl) _ι, 6-diaminohexane (0.554 g '2.22 mmol, 2.00 equivalent) and acetic acid (0.067 g' 1.1 mmol) in anhydrous MeOH 1 0 equivalent) solution was added N- (2-oxoethyl) -4-butylamino-1,8-fluorenyldimethylamine (25.34 g, 85315 -34- 200401034) in 25 ml of anhydrous MeOH. 1.11 mmol). A solution of NaCNBH3 (0.070 g, 1.1 mmol, 1.0 equivalent) in 5 ml of anhydrous MeOH was added to the mixture. The reaction mixture was stirred at 25 ° C for 15 hours. At this point MeOH was removed by rotary evaporation and the residue was dissolved in 30 ml of water. The solution was adjusted to pH 2 with 1 equivalent of HC1 and then stirred at 25t for 1 hour. At this point the solution was adjusted to pH 12 with 1 equivalent of NaOH and then extracted with 3 x 50 ml of CH2C12. The combined organic extracts were washed with 3 × 50 ml of water, dried over anhydrous Na 2 SO 4, dried and concentrated to give a crude colored oil. The crude material was purified by a stone gelatin layer separation method (35 g of gravity-grade gel, 0-50% CH3OH / CH2C12, followed by 45/50/5 of CH3OH / CH2Cl2 / iPrNH2) to yield 0.190 g (32%) of Amine product. FAB MS: calculated C33H45N502 [M] + 544; experimental value [M] + 544. TLC: Mok Silicone 60 plate, Rf 0.42 Observed with 80/20 CH2Cl2 / CH3OH, indanetrione stain and UV (254/3 66). HPLC: HP 1100 HPLC chromatography, Waters 5x100 mm NovaPak HR C18 column, 0.050 ml injection, 0.75 ml / min, 1.5 ml injection ring, detection at 450 nm, A = water (0.1% HFBA) and B = MeCN (0.i% HFBA), gradient_ · 10% B for 2 minutes, 10-80% B for 18 minutes, 80-100% 8 for 2 minutes, 100% 8 for 2 minutes; residence time 17.6 minutes. F. N-2- [6-N- (N-4-dimethylaminobenzyl) -6-N- [2- (5,5-dimethylboryl-2-yl) benzyl ] Aminehexyl]-[2- (5,5-Dimethylpeptan-2-yl) methylene] aminoethyl-4-butylamino-1,8-fluorenedimethylamine will be in 2 ml A solution of (2-bromomethylphenyl) boric acid neopentyl ester (0.390 male 85315 -35- 200401034 g, 1.38 mmol, 5.00 eq) in CHC13 was added to N-2- [6- N- (N-4-dimethylaminobenzyl) aminohexyl] aminoethyl) _cardiobutylamine_ 1,8-naphthalenediamine (0.150 g, 0.276 mmol) and DIEA (0.355 G, 0.478 ml, 2.81 millimoles, 10.0 equivalents). The solution was then stirred at 25 C for 27 hours. At this point the mixture was concentrated and the residue was purified by oxidation column chromatography (100 grams of activated neutral alumina, 0.05% CH3OH / CH2Cl2) to yield 0.024 grams (19%) of a sticky brown oil. FAB MS (Glycerin Matrix): Calculated c53H67B2N508 [M] + 924 (diethylene glycol adduct instead of bis-neopentyl boronic acid); experimental value [M] + 924. TLC: Mok's neutral alumina flat plate, 0.62 with CH2C12 / CH30H at 80/20 and UV (254/366) observation. HPLC. HP 1100 HPLC chromatogram 'Waters 5x100 mm NovaPak HR C18 column' 0.050 ml injection, 0.75 ml / min, 1.5 ml injection ring, detection at 450 nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA) 'Gradient: ΐ〇% β over 2 minutes, 10-80% B over 18 minutes' 80-100% B over 2 minutes, IO% b over 2 minutes; residence time 20.7 minutes. G. N-2- [6-N- (N-4-dimethylaminobenzyl) _6_N- [2- (dihydroxyboryl) benzyl] aminohexyl]-[2- (toluene) Fluorenyl] aminoethyl_4-butylamino-1,8-p-dimethylamine (nBuF-hexyl-Q-bisborate) The free bisboronic acid product used in glucose studies is derived from N_2- [ 6-N- (N-4-dimethylaminofluorenyl) _6_N_ [2- (5,5-dimethylboron alpha benzyl) fluorenyl] aminohexyl]-[2- (5,5-bis Methylborofluoren-2-yl) fluorenyl] amine ethyl-4-butylamine 85315 -36 · 200401034 dimethyl-1,8 -fluorenyldimethyl Si; dissolution of amines in MeOH / PB S buffer system β Η Glucose and lactate were used to determine the fluorescence modulation effect of the marker compound (which includes two discriminating elements) prepared in this example with glucose and lactate. Figure 5 shows the normalized fluorescence emission of a 0.015 mg molecular weight marker compound solution in a) 0/20 mg molecular weight glucose; b) 0-20 mg portioned lactate in 70/30 MeOH / PBS ( 1 / 1〇 @ 535nm). The spectrum was recorded using a Shimadzu rf-5301 spectrophotometer, which was excited at @ 450 nm, with an excitation aperture of 1.5 nm, and a 1.5-micron emission aperture, at room temperature. Error bars are the standard deviations of 3 replicates for each data point. The fluorescence of the marker is affected by the presence of glucose, but rhenium is not substantially affected by the presence of lactate. Example 6 Glucose or lactate pair containing N- [3- (methacrylamino) propyl] -3,4 · diamidyl-9,10-dioxy-2-anthanoic acid amine (Alizarin Red s monomer) and α, α, _bis [N_ [2- (5,5-dimethylboramidine_2-yl) benzyl] -N- [3- (methylpropylamidinyl) propylamine Effect of propylamine gel of [M] -M-xylene (bis-boric acid monomer): A. 3,4-dihydroxy-9,10-dioxy_2 · anthracenesulfonium chloride: 3 Sodium salt of 4-4-peryl-9,10-dioxy_2-cis acid (14 g, 39 mmol) was combined with 30 ml of chlorosulfonic acid and heated to 90 ° C for 5 hours After boiling, the solution was cooled to 0t and poured into 1 () 0 grams of ice. After the ice-melted solution was extracted with CH2Cl2 (3 x 100 ml), the dichloromethane extracts were combined, dried over NaJO4, and evaporated to yield 087 g of solid (66% yield). B. MM methylpropylamido) propyl] _3,4_dihydroxy_9,1〇_dioxy_2_anthracene chloride:, -85315 -37- 200401034 will be 3,4- -9,10-monooxy_2-anthracene chloride (96 mg, 028 mmol) and N- (3-aminopropyl) methylpropanamide hydrochloride (108 Mg, 0.06 mmol) was combined with 20 ml CEbCl2. Et3N (303 mg, 3 mmol) was added to the suspension. The mixture was stirred at room temperature for 24 hours, filtered and the solution was evaporated. The obtained solid was subjected to column chromatography on Si02 (10 g) using CH2Cl2 / MeOH (90/100) as a dissolving agent. The product was obtained as a red solid (80 mg, 64% yield). FAB MS: calculated C21H20N2O7S M + 445; experimental value M + 445. HPLC. HP 1100 HPLC chromatogram, Waters 5x100 mm NovaPak HR C18 column, o.ioo ml injection, 0.75 ml / min, 2 ml injection ring, detection at 370 nm, a = water (0.1% HFBA) And B = MeCN (0.1% HFBA), gradient: i0% b over 2 minutes, 10-80% B over 18 minutes '80 -100% 8 over 2 minutes, 100% 6 over 2 minutes; residence time 17.67 minute. C. cc N- (3-Aminopropyl) methylpropylamine hydrogen in bis [3- (methylpropylamido) propylamino] -i, 4-xylene in 75 ml of anhydrous MeOH Chlorate (3.00 grams, 16.8 millimoles, 2.21 equivalents), DIEA (6.5 grams' 8.8 milliliters, 50 millimoles, 6.6 equivalents), terephthalaldehyde (1.02 grams, 7.60 millimoles) and ^ 28 A solution of 〇4 (10.7 g, 75.3 millimoles, 9.91 equivalents) was stirred at 25 ° C for 18 hours in the dark. Add more Na2SO4 (10.7 g, 75.3 millimoles, 9.91 equivalents) at this time and continue stirring for more than 6 hours. At this time, the solution was filtered, and NaBH4 (1.73 g, 45.7 mmol '6.01 equivalent) was added to the filter in portions and then stirred at 25 ° C for 21 hours. The suspension was filtered through celite and the furnace was concentrated. The residue was dissolved in 85315 -38- 200401034 in 100 ml of CH2Ch and washed with 1 x 25 ml of saturated aqueous NaHC03. The organic extract was dried over anhydrous Na2SO4, filtered and concentrated to give a viscous oil. The reaction was continued as it was. HPLC: HP 1100 HPLC chromatography, Vydac 201TP 10x250 mm column '0.100 ml injection, 2.00 ml / min, detection at 260 nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA) Gradient: 10% B for 2 minutes, 10-80% B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; residence time 15.8 minutes. D. a, a bis [N- [2- (5,5-Dimethylpeptan-2-yl) fluorenyl] -N- [3- (methylpropanamine) propylamine]- 1,4-xylene will be α, α bis [3- (methylpropylamido) propylamine] -1 in 75 ml of CHWh, cardiac xylene (2.94 g, 7.61 mmol), DIE A (2.9 7 g, 4.00 ml, 23.0 mmol, 3.02 equivalents), (2-bromomethylphenyl) borate neopentyl ester (6.50 g, 23.0 mmol, 3.02 equivalents), and 8111 '(5 mg 'As a marker) The solution was stirred at 25 ° C in the dark for 28 hours. At this time, the mixture was washed with 1 x 25 ml of saturated aqueous NaHC03. The organic extract was dried 'over anhydrous NazSCU, filtered and concentrated. 200 ml of ether was added to the residue 'and the suspension was stirred for 18 hours. The suspension was filtered and the residue was dissolved in CH2C12, filtered and the filtrate was concentrated. 150 ml of ether was added to the solid residue, and the suspension was stirred for 18 hours. The suspension was filtered 'at this point to produce 1.98 g (33%) of a fluffy pink powder. FAB MS: Calculated C46H64B2N4 06 [M] + 790; experimental value [M + 1] + 791. HPLC: HP 1100 HPLC chromatography, Waters 5x100 mm 85315 -39- 200401034

NovaPak HR C18 column, 0.050 ml injection, 0.75 ml / min, detection at 280 nm, a = water (0.1% HFBA) and B = MeCN (0.1% HFBA), gradient: 10% b 2 minutes, 10-80% B over 18 minutes, 80-100 ° / 〇B over 2 minutes, 100% b over 2 minutes; residence time 13.4 minutes. E. Contains N- [3- (methacrylamino) propyl] -3,4-dihydroxy-9,10-dioxy-2-anthracenesulfonamide (alizarin red s monomer) and 〇 ;, Α bis [N- [2- (5,5-dimethylboryl p-s-2-yl) benzyl] -N- [3- (methacrylamino) propylamine] _ι, 4- Preparation of acrylamide gel of xylene: Preparation of an ethylene glycol solution containing 30% by weight of propylamidine and 0.8% by weight of N, N'-methylenebisacrylamide. Add N- [3_ (methylpropanamine) propyl] _ 3,4-dihydroxy-9,10-dioxy-2-anthracenesulfonamide (1.5 mg, 3 38 × 10-6 mol ) And α, α bis [N- [2- (5,5-dimethylborofluoren-2-yl) yl] -N- [3- (methylpropylamido) propylamino] -1 4,4-xylene (28 mg, 3.54 x 10.5 mol) was combined with 800 microliters of ethylene glycol monomer solution and 40 microliters of 5% by weight aqueous ammonium persulfate. This formulation was placed in a glove box flushed with nitrogen together with a model constructed of a glass microscope sheet and a 100 micron stainless steel spacer plate. Microliter ^ '-tetramethylethylene diamine aqueous solution 丨 microliter' ^^^^^ was added to the monomer solution to accelerate the polymerization, and the final formulation was poured into a glass mold. The model was left under nitrogen for 16 hours, then it was inserted into pBS (PH = 7.4) 'and the glass lenses were separated to supply a hydrogel polymer in the form of a film. The obtained hydrogel film was washed with 100 ml of sodium sulfate lauryl sulfate containing 100 mg of molecular weight in phosphate-buffered saline for 3 days, and the solution was changed daily, followed by MeOH / PBS (20/80 by volume, 3 x 100 ml) ) Wash and finally wash with PBS (PH = 7.4, 3x100 ml). The hydrogel polymer was stored in 85315-40-40200401034 in PBS (10 mg molecular weight PBS, pΗ = 7.4) containing 0.2% by weight of sodium azide and 1 mg of molecular weight] gDTA sodium salt. F. Joint effect with absorption values of glucose and lactic acid hits The marker hydrogel (which includes two discriminating elements) prepared in this example was used to adjust the absorption values of glucose and lactate. Ethylamine gel was loaded in a PMMA tank in the same manner as described in Example 4. The scaled buffered saline (pBS PH7_4) including a predetermined amount of glucose or sodium lactate was heated to 37 t in a water bath, and placed in a pmma tank including a gel, and then the PMMA tank was allowed to stand at 37. (: Equilibrium is reached in 15 minutes. Each measurement of glucose or lactate concentration is performed 3 times. Each measurement uses the absorption value at 650 nm as a blank value, from all a values (45 ° when micron) And A value (530 nm) minus a (650 nm). Figure 6 shows an acrylamide gel (4 mg molecular weight alizarin with and without glucose and 44 mg molecular weight bisboronic acid monomer acrylamide gel (3 Figure 7 shows the effect of glucose on the absorption value of acrylamide gel (30%) containing 4 g of molecular weight alizarin $ and mg of biboric acid monomer in the molecule. Figure 8 Acid Jun's effect on the absorption value of promethazin gel (30%) including 4 mg molecular weight alizarin§ and 44 mg molecular weight diboronic acid monomer. The absorption value of the marker is affected by the presence of glucose, but it is essentially It is not affected by the presence of lactate. G. Measured by the fluorescence-regulating effects of glucose and lactate ^ Essentially based on the promethazamine gel synthesized in Example 6 (except the use of 1.9 mg of N- [3- Alkylamino) propyl] _3,4_dienyl_9, ι〇 • di Oxy-2-anthracenesulfonamide and 35 g of α, α, · bis [N_ [2_ (5,5_dimethylboramidine-85315 -41-2 -yl) benzyl] -N- [3 -(Methacrylamine) propylamine] -l, 4-xylene]. Fluorescence adjustment effect. Experiment was performed on Shimadzu RF-5301 PC Fluorometer with variable temperature accessory (at 470 亳Micron excitation, 3/10 nanometer pores, high sensitivity). Acrylamide gel was attached to a piece of 45. angle glass lens, and it was glued to a PMMA fluorescent cell. 25 ml of pBS ( pH = 7 4) Put into the tank and heat to 37 C ° to prepare a glucose stock solution (100 mg molecular weight and 500 halo molecular weight) in pBS (pH = 7 4), and heat to 37 in a water bath ° C. Add aliquots of heated glucose stock solution to the PMMA tank periodically, and monitor the fluorescence intensity (measured every 2 minutes) at the time of 550 nm at the same time. Use YSI type 2〇STAT A glucose analyzer was added to measure the glucose concentration in the PMMA tank. The results shown in Figure 9 show that the addition of glucose decreases the fluorescence intensity of the marker hydrogel. The phase was observed in Figure 〖〇 The same effect, which shows the effect of glucose on the fluorescence spectrum of gels of the same type. It is believed that this effect occurs because of the following considerations. The methylalanine monomer (receptor molecule) of alizarin 8. Including nearby diol functionality and monomer functionality (refer to the following structure p. Alizarin S and bis-borate discriminating element monomers (refer to the following structure) can react with each other reversibly in aqueous solution and organic solvent, Borate is formed. The borate molecule formed in this reversible reaction is a fluorescent molecule, but the alizarin S monomer itself is in an aqueous solution and an organic solvent (such as

MeOH) does not visually see fluorescence emission. Therefore, once combined with the glucose discerning element, the alizarin S will change its vortex, w, and its specific properties such as absorption value and fluorescence yield. 85315 -42200401034

It can be used with hydrogel monomers and cross-linking agents to prepare a solution of Alizarin S with monomer functionality and a glucose discrimination element with monomer functionality. The copolymerization of this mixture produces a hydrogel material that can diffuse to a variety of small to medium sized molecules, so it can be said to detect and quantify analytes. Analytes such as glucose can diffuse and displace receptor molecules previously bound to discriminating elements in the hydrogel matrix. This event caused a change in the optical properties of the hydrogel membrane, as it currently includes a larger number of receptor molecules that are not bound to a discriminating element. The fluorescence-regulating effect of the marker compound prepared in this example (which includes two discriminators) on glucose and lactate was also determined. Experiments were performed on a Shimadzu RF-5301 PC fluorometer equipped with a variable temperature accessory (excitation at 47 nm, 5/10 nm pores, low sensitivity attaching acrylamide gel to a piece of 45. On a glass slide, glue it to a pMMA fluorescent cell. Load 2.5 mL of PBS (PH = 7.4) into the cell and heat to 37 ° C in a water bath. Prepare sodium lactate in PBS (pH = 7.4) Stock solution (100 mg molecular weight) 'and heated to 37 ° C in a water bath. Prepare a glucose stock solution (100 mg molecular weight and 500 mg molecular weight) in pBs (pH = 74), and heat in a water bath to 37 ° C. Add an equal amount of heated lactate stock solution to 85315 -43-200401034 into the PMMA tank periodically, and monitor the fluorescence intensity based on time as a function of 550 nm (measured every 2 minutes) 'Until the lactate concentration reaches 8 mg of molecule 1. Then, aliquots of heated glucose stock solution are periodically added to the PMMA tank, while monitoring the ordinary light intensity based on time as a function of 550 millimeters (2 minutes of mother) Measure once) 〇 use ysj [type 2300; § τ AT A glucose analyzer was added to measure the glucose concentration in the PMMA tank. The results shown in Figure 丨 i show that the added lactate has no significant effect on the fluorescence intensity of the marker hydrogel, and the subsequent addition of glucose will reduce the label Fluorescence Intensity of Biohydrogels. Example 7 Single methacrylamide monomer of bis-borate-anthracene:

A. 9-chlorofluorenyl-io-[[2- (2-hydroxyethoxy) ethylamino] methyl] anthracene hydrochloride will be 2- (2-aminoethoxy) ethanol (〇 • 495 g, 0.475 ml, 4.71 millimoles) was added to a suspension of 9,10-bis (chlorofluorenyl) anthracene (5.18 grams' 1 8.8 millimoles, 3.99 chapters) in 200 ml NMP. Put the mixture in the dark

Stir in I for 17 hours. At this time, the reaction mixture was concentrated to ~ 50 ml under reduced pressure at 50 t. The residue was purified by silica gel chromatography (50 grams of gravity-grade silica gel, 0-10% CH3OH / CH2C12) to yield 0.425 grams (24%) of a yellow / orange solid. TLC: Mok Silicone 60 flat plate, Rf 0.72 observed with 70/30 CH2C12 / CH30H, 85315 -44-200401034 with UV (254/3 66), indantrione stain. HPLC: HP 1100 HPLC chromatography, Vydac 201TP 10x250mm column, 0.100ml injection, 2ml / min, detection at 370nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA) Gradient: 10% B for 2 minutes, 10-80% B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; stay time 16.1 minutes.

Β · 9-[[2- (2-hydroxyethoxy) ethylamino] methyl] -10-[[(3-methylassocidinyl) propylamino] methyl] anthracene-will be in 25 ml of CHC13 Of 9-chloromethyl-10-[[2- (2-hydroxyethoxy) ethylamino] methyl], hydrochloride (1.56 um, 4.10 mmol) N- (3-aminopropyl) methacrylamidine hydrochloride (3.08 g, 17.2 mmol, 4.2 equivalents), DIEA (5.19 g, 7.00 ml, 40.1 mmol, 9.8 equiv) and ~ 3 mg of BHT in suspension. The mixture was then stirred in the dark for 92 hours. At this point the reaction mixture was filtered and washed with 2 x 40 ml of NaHC03 (saturated aqueous solution). The organic extract was dried over anhydrous Na2S04, filtered and concentrated to produce a viscous orange solid, which was purified by alumina chromatography (50 g of activated neutral alumina, 0-5% CH3OH / CH2C12), This gave 0.364 g (20%) of an orange solid. 1 ^ (:: Moke Silicone 60 flat plate, 1 ^ 0.16 to 70/3 0 of (: 112 (: 12 / (: 113011, 85315 -45- 200401034 with UV (254/366), indan trione stain Observation: HPLC: HP 1100 HPLC chromatography, Vydac 201TP 10x250 mm column, 0.100 ml injection, 2 ml / min, detection at 370 nm, A = water (0.1% HFBA) and B = MeCN (0.1 ° / 〇HFBA), gradient: 10% B for 2 minutes, 10-80% B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; residence time 16.8 5 minutes.

C. 9- [N- [2- (5,5-dimethylboramidine-2: -yl) benzyl] -N- [3- (methacrylamino) propylamine] methyl]- 1〇- [1 ^-[2- (5,5-dimethylborofluoren-2-yl) benzyl]-: ^-[2- (2-hydroxyethoxy) ethylamino] methyl] Anthracene (mono-methylpropylamidamine monomer) 9-[[2- (2-hydroxyethoxy) ethylamino] fluorenyl] -10-[[(3-methyl Amido) propylamino] methyl] anthracene (0.343 g, 0.763 mmol), 0: 8 (0.965 g, 1.30 ml, '9.8 equivalent), and (2-bromomethylphenyl) neopentyl borate A solution of the ester (1.09 g, 3.85 mmol, 5.0 eq) was stirred at 23 ° C in the dark for 25 hours. At this point the reaction mixture was first evaporated by rotary and then concentrated using a vacuum pump to remove DIEA. The residue was purified by alumina column chromatography (40 g of activated neutral alumina, 0-10% CH3OH / CH2C12) to yield 0.299 g (46%) of yellow-orange solid 85315 -46- 200401034. This compound can be copolymerized with a suitable monomer (as previously described), deprotected and used to detect glucose. FAB MS: calculated C51H65B2N307 [M] + 854; experimental value [M + 1] + 855. TLC: Mock-experimental alumina flat plate, Rf 0.35 observed at 95/5 CH2Cl2 / CH3OH, UV (254/366). HPLC: HP 1100 HPLC chromatography, Vydac 201TP 10x250 mm column, 0 · 100 ml injection, 2 ml / min, detection at 370 nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA), gradient: 10% B over 2 minutes' 10-80% B over 18 minutes, 80-100% B over 2 minutes, 100 ° / 〇B over 2 minutes; residence time 19.7 minutes. Example 8 Bis-borate-anthracene di-methyl% propyl, melamine monomer

A. 9,10-Bis [N- [2- (5,5-Dimethylborofluoren-2-yl) yl] -N- [2- (2-methylpropanyloxyethoxy) Ethylamino] methyl] anthracene, 0.110 9,10-bis [N- [2- (5,5-dimethylborofluoren-2-yl) fluorenyl] -N- [in 5 ml of CH2C12 2- (2-hydroxyethoxy); ethylamino] methyl] anthracene (0.100 g, 0.120 millimoles' Reference Example 2), methacrylic acid (0.112 g 85315 -47- 200401034 ml, 1.30 millimoles, 10.8 equivalents), DCC (0.3 16 grams, 1.53 millimoles, 12.8 equivalents), and N, N-dimethylaminopyridine (0.014 grams, 0.11 millimoles, 0.92 equivalents) were dissolved in 0 Stir at ° C for 1 hour, then at 23 ° C for 22 hours. At this point the reaction mixture was filtered and concentrated by rotary evaporation. The residue was purified by oxidation column chromatography (30 g of activated neutral alumina, 0-20 / 〇CH3OH / CH2C12) to yield 0.030 g (26%) of a yellow solid. This compound can be copolymerized with a suitable monomer (as previously described), deprotected and used to detect glucose. : FAB MS · Calculated C56H7〇B2N2010 [M] + 953; Experimental value [M] + 951 (weak molecular ion peak). TLC: Mok alkaline alumina plate, Rf 0.67 was observed at 95/5 of CH2Cl2 / CH3OH and UV (254/366). HPLC: HP 1100 HPLC chromatography, Waters 5x100 mm NovaPak HR C18 column, 0.100 ml injection, 0.75 ml / min, 2 ml injection ring, detection at 370 nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA), gradient: 10% B for 2 minutes, 10-80% B for 18 minutes, 80-100% B for 2 minutes, L00% B for 2 minutes; residence time 19.6 minutes. Example 9 Double 5-aminopentylbis-borate-anthracene

85315 -48-200401034 A. 9,10-Bis [[5- (t-Boc) _aminopentylamino] methyl] tzine..

Add 9,10-bis (chlorofluorenyl) anthracene (0.28 g, 1 mmol), DIEA (7.0 ml, 40 mmol), mono-tert-butoxycarbonyl, 5-diaminopentyl A suspension of alkane (3.75 g, 10 mmol) and 50 ml of CHCh was stirred at 45 ° C for 2 days in the dark. The solution was washed with saturated H20 / NaHC03, the organic phase was dried (NazSCU), and the solvent was evaporated. The residue was purified by alumina chromatography (40 g of activated neutral alumina, 95/5 vol% CI ^ Ch / MeOH) to yield 0.55 g of a viscous oil. Use the following procedure as it is. % Β · 9,10-bis [N- [2- (5,5-Difluorenylborofluoren-2-yl) amyl] -N- [5- (t-BOC) -aminopentylamino] Methyl] anthracene

'I will be 9,1 0-bis [[5- (t-Boc) _aminopentylamino; 20 methyl] anthracene (0.3 g. 0.49 mmol), DIEA ( A solution of 0.35 ml, .2 mmole) and neopentyl (2-bromomethylphenyl) borate (0.566 g, 2.0 mmole) was stirred at 25 ° C in the dark for 2 days. At this time, the reaction mixture was concentrated in vacuo, and the residue was purified by alumina chromatography (60 g of activated neutral alumina, 98/2 vol% iCH2Cl2 / MeOH) to yield 0.401 g of a yellow oil. This substance was used as it is in the following steps. 85315 -49- 200401034

C. 9,10-bis [N- (2-Dihydroxyborylfluorenyl) -N- [5-aminopentylamino] methyl] anthracene trifluoroacetate will be 9,10-bis [N- [ 2- (5,5-Dimethylboron 4-2-yl) benzyl] -N- [5- (t-BOC) -aminopentylamino] methyl] anthracene (0.4 g, 0.39 mmol ) Dissolved in 20 ml of CH2Cl2 / TFA (80/20 vol%). The solution was stirred for 12 hours, the solvent was evaporated, and the residue was washed with 10 ml. Of ether. A total of 373 mg (72% yield) of solid was obtained. The product has ~ 80% purity (RP-HPLC). This compound can be copolymerized with a suitable monomer (as previously described), deprotected and used to detect glucose. HPLC: HP 1100 HPLC chromatography, Waters 5x100 mm NovaPak 'HR C 18 column, 0.050: ml injection, 0.75 ml / min, detection at 360 nm, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA), gradient: 10% B in 2 minutes! 10-80% B over 18 minutes, 80-100% B over 2 minutes, 100% B over 2 minutes; stay time 16.0 minutes. Example 1 0 85315 -50- 200401034

A · N-2- (tert-butoxycarbonyl) aminoethyl_4-bromofluorenecarbohydrazone: ’’

Add Nt-Boc-Ethylenediamine (Fluka, 16 g, 10 mmol) and 4-Cryl-i, 8-phthalic anhydride (adretrid, 2 77 g , 10 mmol) and 60 ml of absolute ethanol, and the suspension was stirred at 6 (rc for 20 hours' cooled to room temperature and filtered. The obtained solid was washed with 30 ml of cold EtOH I. Wash and Drying under vacuum. 3 84 g (91%) yield. NMR (CDC13): 1.28 (9H, s); 3.52 (2H, t); 4.35 (2H, t); 4.92 (1H,, s); 7.84 (1H , t); 8.04 (1H, d); 8.42 (1Η, d); 8.58 (1H, d); 8.67 (1H, d).

B. N-2- (tert-butoxycarbonyl) aminoethyl_4_ (N'_methylaminoethylethylamino) fluorene · 1,8-dicarboxyfluoreneimine: N-methylethylene Diamine (ι · 48 g, 20 mmol) was combined with 2 ml of 1-methyl-2-pyrrolidone (NMP), followed by addition of N-2- (tert-butoxycarbonyl) amine ethyl-4- Bromhexyl-1,8-dicarboxamidine (0.35 g, 0.845 mmol). 85315 • 51-200401034 Known solution at 45 ° C: mix for 40 hours, then evaporate NMP and N-methylethylenediamine under vacuum. The obtained residue was subjected to column chromatography (20 g of silica gel, first with CH2C12 / MeOH (90/10), and then with CH2Cl2 / MeOH / Et3N (75/20/5)). A yellow solid was obtained (0.311 g, 89% yield). Take Rp_HpLC: check purity.

C. N-Aminoethyl_4_ (n, _Aminovinyl_ν, '-[2 · (Diborane) Uryl] methylamino) Nylidene-1,8-Dibenzyl Amine trigas acetate: N-2- (tert-butoxycarbonyl) aminoethylmethylaminoethylamino) naphthylene-U-dicarboxyamidoimine (0.3 g, 0.73 mmol), 2 Bromomethylphenylboronic acid, pinacol esters (0.6 g, 2 mmol), N, N-diisopropyl-N-ethylamine (1.3¾ liters, 8¾ mols) were combined with 10¾ liters of CH2C12. The solution was stirred for 20 hours, followed by the addition of 2 grams of PS-diamine resin (Argonaut Technogles, 3.38 ¾ mole / gram). The reaction mixture was stirred with the resin for 10 hours, then the resin was removed by filtration, and washed with < CH2Ci2 (2 x 20 ml). The combined CH ^ Cl2 falling liquid was evaporated and dried under vacuum.

• 52- (H〇) 2B 85315 200401034 Dichloromethane including 20% by volume and 5% by volume of triisopropylformamidine was added to the resulting orange residue. The resulting solution was allowed to stand at room temperature for one hour, then the agent was evaporated and the residue was triturated with ether to give a yellow solid. The solid was filtered and dried in vacuo (580 mg yield). The purity of the product was checked by Rp_HPLC. It is used in the following steps as it is. D · N- (3-Dihydroxyboryl_5-nitrophenylphosphoniumamino) ethyl_4_ (N, _aminovinyl_N "-[2_ (dihydroxyboryl) benzyl] methyl Amine) fluorene-1,8-dicarboxyfluorenimine: N-aminoethyl-fluorene- (N'-aminoethylfluorenyl_n, '-[2- (dichloroboryl) yl) ] Methylamino) stilbene-1,8-dicarboximide trifluoroacetate (0,225 g, 0.4 mmol), 3-carboxy-5-nitrophenylboronic acid (0.085 g, 0 · 4 mmol), diphenyl azide phosphate (0.13 ml '0.6 mmol) and 2 ml of anhydrous DMF. Add N, N-diisopropyl-N-ethylamine (0.7 ml, 4 mmol) Ear), and stir / 4 liquid: Stir for 20 hours. Add acid (10 ml) to the reaction mixture, separate the insoluble residue and sonicate with 5 ml of CH2C12 to produce an orange solid, filter it and Dry under vacuum (38 mg, 15% yield). Check the purity of the solid by RP-HPLC. NMR (dmso-d6 / D20, 90/10): < 5 2 · 32 (3Η, s); 2.82 (2H , t); 3.58 (2H, t); 3.65 (2H, t); 3.7〇 (2H, s); 6.65 (1H, d); 7.〇-7.3 (4H, m); 7.68 (1H, t) ; 8.18 (1H, d); 8.42 (1H, d); 8.47 (1H, d); 8.1-8.35 (3 H, m). E. N- (3-Dihydroxyboryl-5-nitrophenylphosphoniumamino) ethyl-4- (N'-aminovinyl-N ,,-[2 -(Dihydroxyboryl) fluorenyl] methylamino) fluorene-M-dicarboxyfluorenimine and glucose interaction test 85315 -53- 200401034 in MeOH / phosphate buffered saline (PBS, The molecular weight was 10 mg, ρΗ = 7.4.) The experiment was performed. N- (3-Dihydroxyboryl-5-nitrophenylamido) ethyl-4 in MeOH / PBS (50/50 vol%) The concentration of-(N'-aminoethylamido-N "-[2- (dihydroxyboryl) fluorenyl] methylamino) fluorene-1,8-dicarboxyfluorenimine is a molecular weight of 15 grams. The glucose concentration changed from 0 mg molecular weight to 50 mg molecular weight, and the sodium L-lactate concentration changed from 0 mg molecular weight to 7 mg molecular weight. Experiments were performed on a Shimadzu RF-5301 ’PC fluorometer: the excitation wavelength was set at 430 nm, the emission was monitored in the range of 480-650 nm, 3 / 1.5 nm pore width, and high-sensitivity PMT. The results are shown in Figures 12 and 13, which show that the marker fluorescence of this example is affected by the presence of glucose but not by the presence of lactate. Example 41 6- (cyclohexanecarboxamido) hexylamine marker monomer

Α · 9- [N- [3- (Methylpropylamido) propylamino] methyl] -10-N-[(6-aminohexyl 85315 -54- 200401034 amino) methyl] anthracene 9 , 10-bis (chloromethyl) anthracene (0.150 g, 0.545 mmol) with 3-aminopropylmethacrylamide (0.775 g, 5.45 mmol, 10.0 eq) in 200 ml of CHC13 -(6-aminohexyl) tert-butyl carbamate (1.18 g, 5.45 mmol, 10.0 eq) and some BHT crystals in solution. The reaction mixture was then stirred at room temperature in the dark for 4 days. At this point CHC13 was evaporated and the residue was dissolved in 100 ml of ether. The organic layer was extracted with 8 x 125 ml of saturated aqueous NaHCCb and 5 x 200 ml of phosphate buffer (0.4 g molecular weight, pH 7.0). The pH of the combined phosphate buffer washings was adjusted to pH 11 by adding Na2C03 (saturated aqueous solution), and then extracted with 5 x 300 ml of CH2C12. The combined organic layers were concentrated and the residue was dissolved in 5 ml of a 20% TFA solution in CH2C12. The mixture was stirred at room temperature for 2 hours. At this point the reaction mixture was extracted with 4 x 10 ml of saturated aqueous NaHC03. The pH of the combined aqueous layers was adjusted to pH 11 by the addition of Na 2 CO 3 (saturated aqueous solution), followed by extraction with 4 × 75 ml of CH 2 C 12. The combined organic extracts were dried over anhydrous Na2SO4, filtered and concentrated in vacuo to yield 0.068 g (27%) of the product. TLC ·· a) Before deprotection of Mok Silicone 60 flat plate, Rf 0.16 is observed with 70/30 CH2C12 / CH30H and UV (254/366); the final product is Rf 0.27 with 85 / 14.5 / 0.5 CH2Cl2 / CH3OH / iPrNH2, observed under UV (254/366). HPLC: HP 1100 HPLC chromatography, Waters 8x100 mm NovaPak HR C18 column, 0.100 ml injection, 0.75 ml / min, 0.400 ml injection ring, 360 nm detection, A = water (0.1% HFBA) and 85315 -55- 200401034 B = MeCN (0.1% HFBA), gradient: 10% B for 2 minutes, 10-80 ° / 〇B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; Length of stay 15.5

B. 9- [N- [3- (methylpropylalamido) propylamino] methyl] -10- [N- [6- (cyclohexanecarboxyamido) hexylamino] methyl] Anthracene A solution of cyclohexanecarboxylic acid N-hydroxysuccinimide (0.845 g, 3.76 mmol, 1.03 eq.) Was added dropwise over 9 hours to 9- [N- [3- (Methacrylamino) propylamino] methyl] -10-N-[(6-aminohexylamino) methyl] anthracene (1.68 g, 3.63 mmol) and a few BHT crystals Of solution. The reaction was then stirred at room temperature in the dark for 16 hours. At this point the reaction mixture was concentrated in vacuo and the residue was dissolved in 105 ml of a solution of ether / CH2C12 (90/15). The organic layer was extracted with 4 x 225 ml of phosphate: phosphate buffer (0.4 g molecular weight, ρ 分子量 7.0). The pH of the combined phosphonate buffer was adjusted to ρ: Η11 by adding Na2CO3 (saturated aqueous solution), followed by extraction with 6x500 ml of CH2C12. The combined organic layers were dried over anhydrous Na2S04, filtered and concentrated in vacuo to give 1.2 g (60%) of the product. TLC: Mok Silicone 60 flat plate, Rf 0.30, CH2Cl2 / CH3OH / iPrNH2 of 85 / 14.5 / 0.5, observed under UV (254/366). HPLC: HP 1100 HPLC chromatogram, Waters 8x100 mm] ^ < ^ & 卩 & 1 ^ 101 (: 18-column, 0.100 ml injection, 0.75 ml / min, 85315 -56- 0.400 ¾ injection ring , Detected with 360 nm, eight = water (0., 1% 11.8) and B = MeCN (0, 1% HFBA) 'gradient: 10. 10. 2-8 minutes after the iron is earned 18 minutes, 80-100% B after 2 minutes, 100% / 2 minutes; 174

C. 9- [N- (2-Diphenylborylbenzyl) _N_ [3. ((Methylpropylalanino) propyl) amino] methyl] -10- [N- (2-Dihydroxyborylfluorene Cyclohexanecarboxamido) hexylamino] methyl] Hex. 9- [N- [3- (methacrylamido) propylamino] methyl] -10 in 30 liters of CHCI3 -[N- [6- (Cyclohexanecarboxy! Amino) hexylamino] methyl] anthracene (10 g, 1.8 mmol), DIEA (1.81 g, 2.44 mL, 14.0 mmol, 7.8 A solution of 2-bromofluorenylphenylboronic acid pinacol ester (2.14 grams, 7.20 millimoles' 4.0 equivalents) and a few BHT crystals was stirred at room temperature in the dark for 60 hours. The reaction mixture was concentrated at this time, and the residue (9_ [n_ [2_ (4,4,5,5-tetrafluorenyl-1,3,2-dioxorimido) fluorenyl] -N- [3 -(Fluorenylpropene: enaminoamino) propylamino] methyl] -10-[> 1- [2- (4,4,5,5-tetramethyl-1,3,2-dioxyboron (M) Benzyl] -N- [6- (cyclohexanecarboxamido) hexylamino.] Methyl] anthracene) was suspended in 150 ml of ether. The organic layer was washed with 4 x 50 ml of phosphate buffer (0.4 g molecular weight, pH 7.0). The organic layer was concentrated and the residue was dissolved in 200 ml of 0.1 equivalent of ether in aqueous HC1. The aqueous layer was washed with 3x50 ml of ether: cool acid 85315 -57- 200401034 ethyl acetate (1: 1), and the pH was adjusted to pH 11 by adding Na2C03 (saturated aqueous solution), followed by extraction with 3x150 ml of CH2C12. The combined organic layers were dried over anhydrous Na2S04, filtered and concentrated in vacuo to give a red oily compound. The residue was dissolved in ether and concentrated in vacuo to give 1.17 g (85%) of the product as a yellow solid. TLC: Mok Silicone 60 flat plate, Rf 0.59 with 80/20 CH2C12 / CH30H and UV (254/366) observation. HPLC: HP 1100 HPLC chromatography, Waters 8x100 mm NovaPak HR C18 column, 0.100 ml injection, 0.75 ml / min, 0.400 ml injection ring, 360 nm detection, A = water (0.1% HFBA) and B = MeCN (0.1% HFBA), gradient: 10% B over 2 minutes, 10-80% B over 18 minutes, 80-100% B over 2 minutes, 100% B over 2 minutes; residence time 19.6 minutes. NMR (9: 1 d6-acetone / D20): < 5 0.90 (m, 2H), 1.03 (m, 2H), 1.18-1.30 (m, 6H), 1.3 5-1.48 (4H), 1.62 (m , 1H, 0 = C- CH (CH2) CH2), 1.66-1.75 (m, 7H), 1.77 (m, 2H, N-CH2-CH2-CH2-N), 2.52 (m, 2H, N-CH2- CH2-), 2.63 (m, 2H, N-CH2-CH2-), 2.98 (m, 4H, -CH2-NH-C = 0), 3.98 (s, 4H, benzene-CH2-N), 4.57 (s , 2H, anthracene-CH2-N), 4.59 (s, 2H, anthracene-CH2-N), 5.20 (t, 1H, J = 1.5Hz, C = CH2), 5.46 (s, 1H, C = CH2), 7.4-7.5 (m, 8H, Ar-H), 7.52 (m, 2H, Ar-H), 7.95 (m, 2H, Ar-H), 8.23 (m, 4H, Ar-H). D. With 9- [N- (2-dihydroxyborylfluorenyl) -N- [3- (methacrylfluorenylamino) propylamino] fluorenyl] -1O- [N- (2-dihydroxy Borylfluorenyl) -N- [6- (Cyclohexanecarboxamidin 85315 -58- 200401034 based) Hexylamino] methyl] anthracene N, N-dimethylacrylamide hydrogel prepared in phosphoric acid A solution of N, N-dimethylpropanamide (40% by weight) and N, N, -methylbispropionamine (08% by weight) in a salt buffer (ρΗ = 7.4, 200 mg molecular weight) . 9- [N- (2-Dihydroxyborylfluorenyl) _N_ [3_ (methacrylfluorenylamino) propylamino] methyl] -10- [N- (2-Dihydroxyborylfluorenyl) · N_ [6_ (cyclohexylpyridinylamino) hexylamino] methyl] anthracene (18 mg, 2.15 x 10.5 moles) and 60 mg of fructose were combined with 2 ml of MeOH. This solution was sonicated until all fructose was dissolved and then evaporated to produce a solid. One milliliter of phosphate buffer solution including the monomer was added to the solid. After sonication for 10 minutes, the solution was filtered through a 0.2 micron PTFE membrane filter paper. Aqueous ammonium persulfate (20 microliters, 5% by weight) was combined with the formulation. The resulting solution was placed in a glove box flushed with nitrogen. An N, N, N ,, N, -tetramethylethylenediamine aqueous solution (40 microliters, 5% by weight) was added to the monomer formulation to accelerate the polymerization. The resulting formulation was poured into a model constructed with a glass microscope sheet and a 100 micron stainless steel spacer plate. After maintaining under nitrogen for 8 hours, the model was placed in phosphate-buffered saline (ρΗ = 7.4), the microscope sheet was separated, and the hydrogel was removed. The hydrogel was washed with 100 ml of a molecular weight of sodium lauryl sulfate and 1 mg of EDTA tetrasodium salt in 100 ml of phosphate buffered saline (PBS) for 3 days. The solution was changed daily, and pBS (20/80 '3x100 ml by volume) was washed and finally washed with PBS (pH = 74, 3 x 100 ml). The resulting hydrogel film was stored at a weight including 0.002. Sodium azide and J mg molecular weight EDTA tetrasodium salt in PBS (pH = 7.4). E · Determination of the fluorescence modulation effect of 6_ (cyclohexanecarboxamido) hydroxylamine label 85315 -59 · / DMA hydrogel film prepared with glucose and lactate by the fluorescence regulation effect of glucose . Figure 丨 Dou Zhan does not include 0 to 20 mg of molecular weight a_D_glucose; 0 to 10 mg of molecular weight L-lactate sodium and 0 to 20 mg of molecular weight α-D-glucose in PBS Relative fluorescence emission (1 @ 43 nm) of a hydrogel film of medium (ph7.4, including 0.02% NaN3 & i mg molecular weight EDTA). A hydrogel membrane (100 micron thickness, 8 mm diameter disc) was loaded at 45 °. Corner of the PMMA slot tube. At 37t, all measurements were performed in a Shimadzu ruler ... Fluorometer, with low PMT sensitivity excitation at 370 nm (pore-3¾ μm) and emission at 430 ¾ (pore = 3 nm) ). Use ysi type 2300 STAT plus glucose analyzer! Check glucose and L_sodium lactate concentrations. Error bars are the standard deviations of 3 replicates for each data point. Fluorescence is affected by the presence of glucose 'but not by the presence of lactate. Moreover, the presence of urate (4 mg molecular weight) had no significant effect on the glucose calibration curve of 0-20 mg molecular weight. Example 12 2- (Ethyl) amine label monomer

Chemical name: 9- [N- (2-Dihydroxyborylfluorenyl) -N- [3- (methacrylfluorenylamino) propylamino] methyl] -10- [N- (2-Dihydroxyboron Benzyl) _N_ [2_ (carboxyethyl) amino] 85315 -60- 200401034 methyl] anthracene (unblocked) Chemical formula: C40H45B2N307 MW: 701.4 Physical appearance: solubility of dark yellow powder. PBS / methanol, methanol, Ethanol, chloroform, dichloromethane, blocked markers: 9- [N- [2- (4,4,5,5-tetramethyl-1,3,2-dioxoboryl) benzyl] -N -[3- (methylpropylamido) propylamino] methyl] · 10- [Ν_ [2 · (4,4,5,5_tetramethyl-1,3,2 · dioxoborofluoride ) Benzyl] -N- [2- (carboxyethyl) amino] methyl] anthracene I. Synthesis

A. 9- [N- [3- (methylpropylamido) propylamine;] methylterbutoxycarbonyl) ethylamino] methyl] anthracene

9,10-Bis (chloromethyl) anthracene (5.00 g, 18.2 mmol) was added to 3-aminopropylmethacrylamide (12.9 g, 90.7 mmol) in 700 ml CHCU. 4.99 equivalents ), Cold tert-alanine 3.2 g, 90.9 mmol, 5.0 equivalents) and some BHT crystal winter solution. The reaction mixture was then stirred at 30 ° C for 88 hours in the dark. At this time CHC13 was evaporated and the residue was dissolved in 500 ml of ether. The solution was stirred for 1 hour, at which time salts were precipitated from the solution. The ether solution was filtered and then extracted with 10 x 350 ml of saturated aqueous NaHC03. The ether layer was further extracted with 6 x 350 ml of phosphate buffer (0.2 g molecular weight, pH6.5). Adjust the pH bbb 85315 -61-200401034 of the combined phosphate buffer washing solution to pH 11-12 by adding Na2C03 (saturated aqueous solution), followed by extraction with 6x500 ml of CH2C12. The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo to give a crude oily product. The crude product was purified by a crushed gel layer separation method (50 g of flash-grade silica gel, 0-5% MeOH / CH2Cl2 step gradient) to yield 2.04 g (23%) of a viscous yellow solid. Ding 1 ^: Mok silicone 60 flat plate, 1 ^ 0.29 to 90/10 (: 112 (: 12 / (: ^ 13〇1 ^, UV (254/366) and indan trione dyeing agent to observe. HPLC conditions: HP 1100 HPLC chromatogram, Waters 5x100 mm NovaPak HR C18 column, 0.100 liter injection, 0.75 ml / min, 1_500 ml injection ring, detection at 280 nm, A = water (0.1% HFBA) And B = MeCN (0.1% HFBA), gradient: 10 ° / 〇B for 2 minutes, 10-80% B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; during stay Between 17.0 minutes.

Β · 9- [N- [2- (4,4,5,5-tetramethyl-1,3,2-dioxorimido) benzyl] -N- [3- (methylpropanamide (Propyl) propylamino] fluorenyl] -1〇 _ [] ^-[2- (4,4,5,5-tetramethyl-I, 3,2-dioxorimido) methyl]] N · [ 2- (tert-butoxycarbonyl) ethylamino] methyl] anthracene

I 9- [N- [3- (Methacrylamido) propylamino] methyl] -10- [N- [2- (tert-butoxycarbonyl) ethylamino] in 50 ml of CHC13 Methyl] anthracene (1.5 g, 85315 -62- 200401034 = mole), DIEA (3.16 g, 4.26 ml, 24.4 mmol, 79 detonated, · Tian J, 2-bromomethylphenyl When a solution of pinacyl borate (3.64 g, 12.2 mmol, 3.9 equivalents) and a few BHT crystals was stirred at room temperature in the dark 6] At this time, the reaction mixture was concentrated and the residue was suspended In 2000 liters of ether. The ether layer was extracted with 3 x 125 ml of phosphate buffer (0.2 g molecular weight, pH 7.0), dried over anhydrous Na 2 SO 4, filtered, and concentrated in vacuo to give a crude product. The residue was treated with Wet trituration with hexane gave 214 g (76%) of a white solid. HPLC: HP 1100 HPLC chromatogram, Waters 5 x 100 mm NovaPak HR C18 column, 0. 200 ml injection, 0.75 ml / min, 1.500 ¾ liter injection ring 'measured at 280 ¾ micron, α =: water (Qi% hfbA) and B = MeCN (0.1% HFBA)' Gradient: ι〇% β over 2 minutes, ι〇_80% B through 18 Points I ‘80 -100% B after 2 minutes’ 1 00% β after 2 minutes; stay time 19.2

C. 9- [N- (2-Dihydroxyborylbenzyl) -N- [3- (methacrylamino) propylamino] methyl] -10- [Ν- (2-dihydroxyboryl Fluorenyl) -N- [2- (carboxyethyl) amino] methyl] anthracene will be 9- [N- [2- (4,4,5,5 -tetramethyl) in 5 ml of 20% TFA / CH2C12 Methyl-1,3,2-ditrophic greenhouse flavor) benzyl] -N- [3- (ψpropylpropylamido) propylamino] methyl] · 10- [Ν- [2- (4, 4,5,5-tetramethyl-l, 3,2-dioxorane) benzyl terbutin 85315 -63- 200401034 oxycarbonyl) ethylamino] methyl] (0.294 g, 0.319 mmol Mol) solution was stirred at room temperature in the dark for 22 hours. At this point the reaction mixture was concentrated and the residue was triturated with ether. The residue was dissolved in 5 ml of acetone / water (90:10) and stirred for 2 hours. At this time, the reaction mixture was concentrated, and the residue was triturated with water and PBS (pH 7.4, including 0.02% NaN3 and 1 mg molecular weight EDTA) to obtain 0.062 g (28%) of a light yellow solid recovered. HPLC: HP 1100 HPLC chromatogram, Waters 5x100 mm! ^ 〇 \ ^? &Amp; 1 <: 1111 (^ 18 column, 0.100ml injection, 075ml / min, 1.500ml injection circle ' Detected at 280 nm, a = water (0.1% HFBA) and B = MeCN (0.1% HFBA) 'gradient: ΐ〇% β over 2 minutes, ι〇_80 ° / 〇Β over 18 minutes' 80- 100% B for 2 minutes, 100% b for 2 minutes; residence time 17.4 minutes. FAB MS. Glycerol matrix; calculated C46H53B2N307 (double glycerol adduct) [M] + 813; experimental value [M + 2] + 815. D. It has 9- [N- (2-dihydroxyborylbenzyl) _N_ [3_ (methylpropylamido) propylamino] methyl] -10- [N- (2-dihydroxy Borylbenzyl) _N_ [2_ (carboxyethyl) amino] methyl] anthracene N, N-dimethylpropanamide hydrogel prepared in phosphate buffer solution (pH = 7.4, 200 mg molecular weight) A solution of N, N_dimethylpropanamide (40% by weight) and >1,; ^, _ methylenebispropionamine (0.8% by weight). 9- [N- ( 2-Dihydroxyborylbenzyl) _N_ [3_ (fluorenylpropylalanino) propylamino] methyl] -10- [N- (2-Dihydroxyborylbenzyl) -N_ [2 · (carboxyl Ethyl) amino] methyl] E (14 mg 2.0 × 10 · 5 mole) and 60 mg of fructose are combined with 2 ml of MeOH. This solution is sonicated until all fructose is dissolved and evaporated to produce a solid. 1 ml of phosphate buffer solution including monomer is added to 85315- 64- 200401034 in solid. After sonication for one minute, the solution was filtered through a 0.2 micron PTFE filter paper. Aqueous ammonium persulfate (20 microliters, 5% by weight) was combined with the weekly formulation. The resulting solution was placed in a glove box flushed with nitrogen. Aqueous N, N, N ', N'-tetramethylethylenediamine solution (40 microliters, 5% by weight) was added to the single fa week formulation' to accelerate Polymerization. The resulting formulation was poured into a model constructed with a microscope sheet and a 100 micron stainless steel spacer plate. After maintaining for 8 hours under nitrogen, the model was placed in phosphate-buffered saline, 0.00 mg molecular weight 'pH = 7.4), separate the microscope sheet and remove the hydrogel. The hydrogel was washed with 100 ml of 1 mg molecular weight lauryl sulfate sodium salt and 1 mg molecular weight EDTA tetrasteel salt in phosphate buffered saline (pBS) for 3 days, and the solution was changed daily, followed by EtOH / PBS (20/80 by volume, 3 x 100 ml) and finally washed with pBS (pH = 74, 3 x 100 ml). The obtained hydrogel film was stored in PBS (10 mg molecular weight, pΗ = 7.4) containing 0.02% by weight of sodium azide and i mg of molecular weight EDTA tetrasodium salt. II. Fluorescence-regulating effect with glucose The fluorescence-regulating effect of 2- (carboxyethyl) amine-labeled / DMA hydrogel film prepared in this example with glucose and lactate was measured. FIG. 15 shows in PBS (ρΗ7.4, 0,10 mg molecular weight glucose in the presence of 0 to 10 mg molecular weight α-D_glucose; 0 to 10 mg molecular weight sodium L-lactate; Relative fluorescence emission (1 @ 430 nm) of a hydrogel film including 0.02% NaN3 and 1 mg molecular weight EDTA). A hydrogel membrane (100 μm thickness, 8 mm diameter disc) was loaded at 45 °. Corner of the pMMA slot tube. All measurements were performed in a Shimadzu RF-5301 fluorometer at 37 C, with excitation at 370 nm (porosity = 3 nm) and emission of 43 ° 85315 -65- 200401034 meters at low PMT sensitivity (Pore = 3 nm). The YSI type 2300 STAT plus glucose analyzer was used to check the glucose and sodium L-lactate concentrations. The data are plotted as the average of 3 replicates for each data point. Fluorescence is affected by the presence of glucose, but not by the presence of lactate. Furthermore, the presence of lactate (4 mg molecular weight) has no significant effect on the 0-20 mg molecular weight glucose calibration curve. Example 1 3 Fluorescent glucose marker including two detectable moieties:

85315 -66- 200401034 Chemical name: 9- [N- (2-Dihydroxyboryl: fluorenyl) -N- [3- (methylpropylamido) propylamino] methyl] -1〇- [ Ν- (2-Dihydroxyborylbenzyl) -N- [3- (N-6- (9-anthracenecarboxamido) hexylaminocarbonyl) ethylamino] methyl] anthracene (unblocked) Chemical formula ： C73h87b2n5o7 MW ： 1168 Physical appearance: Degree of decomposition of dark yellow powder. PBS / methanol, methanol, ditol, chloroform 'dichloromethane, and compounds blocked with pinacol; 9- [N- [2- (4,4 , 5,5-tetramethyl-1,3,2-dioxoboryl) benzyl] -N- [3- (methylpropylalanino) propylamino] methyl] · 10_ [Ν_ [2 -(4,4,5,5-tetrafluorenyl-1,3,2-dioxofluorenyl) fluorenyl]-] ^-[3-(> 1-6- (9-anthracenecarboxamido) ) Hexylaminocarbonylethylaminomethyl) Anthracene I. Synthesis

A. 9-Anthracene chloride E-9-carboxylic acid (1.2 g, 5.4 x 10.3 mol) was combined with 15 ml of thionyl chloride. The solution was refluxed for 2 hours' followed by evaporation of the volatile components. The obtained solid was dried under high vacuum for 24 hours to produce 1 > 3 g of substance (quantitative yield). This substance was used as it is in the following steps.

85315 -67- 200401034 BN- (6-Aminehexyl) anthracene-9-renylamine hydrochloride: Add 9-anthracene chloride (1.3 g, 5.4 mmol) in 50 ml of anhydrous CH2C12 dropwise 11.6 grams (100 millimoles) of hexamethylenediamine in 100 ml of CH2C12 at 0 ° C. The solution was stirred at Ot: for 1 hour, then allowed to warm to room temperature and stirred overnight. The solvent was evaporated and 200 ml of water was added to the residue. The mixture was sonicated and stirred for 1 hour and then filtered through *. The filtered solid was dried under vacuum for 24 hours. MeOH (50 mL) and 2 mL of concentrated HC1 were added to the solid, and the MeOH was evaporated. The resulting solid was washed with heat <: 112 (: 12 /) ^ 011 (90/10 vol%) and recrystallized from MeOH to yield 0.51 g (26%) of the product. The product purity was checked by HPLC. HPLC: HP 1100 HPLC chromatography, Waters 5x100 mm NovaPak HR C18 column, 0.1 ml injection, 0.75 ml / min, 2 ml injection ring, detected at 280 nm, A = water (0.1% HFBA) and B = MeCN (0.1 % HFBA), gradient: 10% B over 2 minutes, 10-80% B over 18 minutes, 80-100% B over 2 minutes, 100% B over 2 minutes; residence time 16.5 minutes.

85315 -68- 200401034 C. 9- [N- [2- (4,4,5,5-tetramethyl-i, 3,2-dioxorimido) fluorenyl] _N_ [3_ (methacrylic acid) Amine) propylamino] methylΗ〇_ [Ν_ [2_ (4,4,5,5_tetramethyl_1,3,2-dioxofluorenyl) fluorenyl] -N- [3- (Ν -6- (9-anthracenecarboxamido) hexylaminocarbonylethylaminomethyl) anthracene: 9- [N- [2- (4,4,5,5-tetramethyl-fluorene, 3, 2-dioxorimido) methyl] · ν · [3- (methacrylamino) propylamino] methyl] _10_ [Ν_ [2_ (4,4,5,5_tetramethyl_ 1, 3,2-monooxy butterfly flavor) Ethyl] -N- [2-Acetylamido] methyl] Ci (40 mg, 4.5x10 5 moles) and N- (6-aminohexyl) anthracene- 9-Carboxamide hydrochloride (20 mg '5.6 x 10-5 moles), diphenyl azide phosphate (15.4 mg, 5.6 x 10-s mole) and 2 ml of DMF were combined. Diisopropylethylamine (20 microliters, 2 44 x 10-4 moles) was added to the mixture 'and the solution was stirred at room temperature for 24 hours. DMF was evaporated under high vacuum, and the residue was dissolved in 50 ml of Eto-Ac and washed with water (3 x 10 ml). The EtOAc solution was separated, dried (Na2SO4) and evaporated to give 46 mg (87% yield) of a solid. The purity of the material was checked by HPLC. HPLC: HP 1100 HPLC chromatogram, Waters 5x100 mm NovaPak HR C18 column '0.1 ml injection, 0.75 ml / min, 2 ml injection ring' detected at 280 nm, a = water (〇.i% HFBA) And B = MeCN (0.1% HFBA), gradient: 10 ° / 〇B for 2 minutes, i〇_80% B for 18 minutes, 80-100% B for 2 minutes, 100% B for 2 minutes; stay time 20.42 minute. FAB mass spectrometry: glycerol matrix; calculated C67H75B2N509 (diglycerol adduct) [M] + = 1116; experimental value [M + 1] + = 1117. D. Fluorescent effect of glucose on the markers fixed in the hydrogel membrane 85315 -69- 200401034 Preparation of HEMA / methylpropionate hydrogel with glucose markers: Prepared in phosphate buffer solution (pH = 7.4, 200 mg molecular weight) 50% by weight of 2-hydroxyethyl methacrylate (4.75 g) and methacrylic acid (0.25 g). Its solution. Glucose markers (11 mg, 0x10. 5 mol) and 60 mg of fructose were combined with 2 ml of MeOH. This solution was sonicated until all fructose was dissolved and evaporated, and a solid was produced. One milliliter of phosphate buffer solution including the monomer was added to the solid. After sonication for 10 minutes, the solution was filtered through a 0.2 micron PTFE filter paper. Aqueous ammonium persulfate (20 microliters, 5% by weight) was combined with the formulation. The resulting solution was placed in a glove box flushed with nitrogen. N, N, N ,, n, -tetramethylethylenediamine aqueous solution (40 microliters of 5 mils A) was added to the monomer | week formulation to accelerate the polymerization. The resulting formulation was poured into a model constructed with a microscope plate and a 100 micron stainless steel spacer plate. After maintaining under nitrogen for 8 hours, the model was placed in phosphate-buffered saline (10 mg molecular weight, ρΗ = 7.4), the microscope slide was separated, and the hydrogel was taken out. The hydrogel was washed with 100 milliliters of 1 mg molecular weight lauryl sulfate sodium salt and 1 mg molecular weight EDTA tetrasodium salt in phosphate buffered saline (PBS) for 3 days, and the solution was changed every day, followed by EtOH / pBS (20/80 '3 x 100 ml by volume) and finally pBS (pH = 74, 3 x 100 ml). The resulting hydrogel film was stored in pbs (10 mg molecular weight, pΗ = 7.4) including 0.02% by weight sodium azide and 1 mg molecular weight EDTA tetrasodium salt. Effects of Glucose and Sodium L-Lactate on Hydrogel Films Including Glucose Markers 85315 • 70- 200401034 experiments were performed on a Shimadzu RF_53001 PC Fluorometer equipped with a variable temperature accessory. The excitation wavelength is set at 370 nm, 3/3 nm pores, low PMT sensitivity, and scanning emission from 400 to 600 nm. The YSI type 2300 STAT plus glucose analyzer was used to check glucose and L-lactate concentrations. The hydrogel membrane (100 micron thickness, 8 mm diameter circle) was loaded into a PMMA tank tube at an angle of 45 °. Phosphate-buffered saline (PBS, pH = 7.4) including the expected amount of glucose, sodium L-lactate, and glucose with sodium L-lactate was heated to 37 ° C in a water bath and placed in a container including hydrogel PMMA slot. After each addition, the PMMA bath was allowed to equilibrate at 37 ° C for 45 minutes. Fluorescence intensity measurements of each glucose / lactate concentration were performed on two different samples, and the average value was used in the calibration curve. Calibration curves were obtained for glucose, sodium L-lactate, and glucose in the presence of 3 mg of molecular weight sodium L-lactate (fluorescent intensity versus concentration at 430. nm). The results are shown in FIG. 16. Real Θ 14 6-Π-carboxypropylamido) hexylamine-based labeling monomer

Compounds blocked with pinacol: 85315-71-200401034 9- [N- [2- (4,4,5,5-tetrafluorenyl-1,3,2-dioxofluorenyl) benzyl] -N -[3- (methylpropylamido) propylamino] methyl] _1- [1 ^-[2- (4,4,5,5-tetramethyl-1,3,2-dioxo Boron) benzyl] -N- [6- (3-carboxypropylamido) hexylamino] methyl] anthracene unblocked compound: 9- [N- (2-Dihydroxyborylbenzyl)- N- [3- (methylpropylalanino) propylamino] methyl] -10- [N- (2-dihydroxyborylbenzyl;) _ N- [6_ (3_carboxypropylamidoamine) Hexylamino] fluorenyl] anthracene synthesis. Synthesis can be performed in a manner similar to that of Example 11, which uses 9_ [n_ [3- (methacrylamino) propylamino] methyl] i__n_ [ 6- (hexylamino) fluorenyl] sense as a raw material. In contrast, the amine raw material and the monomethyl acetic acid of N-methylsulfonyl imine (NHS, propyl # 丄 a «) 酉 (instead of. Cyclohexanecarboxylic acid used in Example 11) NHS ester of acid). Hail | Yu therefore requires an external alkaline hydrolysis step to complete the synthesis

85315 -72- 200401034 Example 1 5 Glucose label / monomer excited by visible light

Chemical name: N- (3-methylpropylamidopropyl) -4- [2-N-[[2- (dihydroxyboryl) benzyl]-[6- (N- [2- ( Dihydroxypyridyl) fluorenyl] -6-N- (3-carboxypropanidine: aminoethyl) aminohexyl)] aminoethylamino] fluorene-1,8-dicarboxyfluorenamine Chemical formula: c47h6. b2n6o1 () MW: 890 can be synthesized as shown below: 85315 73- 200401034

B (OH) 2 85315 -74- 200401034 Example 16 Excited glucose marker / monomer replaced with visible light

Chemical name: N-butyl-4- [2-N-[[2- (dihydroxyboryl) benzyl]-[6- (N- [2- (dipentanyl) amyl] -6- N- (2-Methyldiaminoaminoethyl) aminohexyl)] aminoethylamino] naphthyl-1,8-dicarboxyamido Chemical formula: c44h57b2n5o7 MW: 789.5 Compounds can be synthesized as shown below: 85315 75- 200401034

85315 -76- 200401034 Brief description of the figure Figure 1 illustrates the normalized fluorescence emission (1 / 1〇 @ 420nm) of the marker as described in Example 1. Figure 2 illustrates the normalized fluorescence emission (1 / 1O @ 428 nm) of the marker as described in Example 2. Figure 3 illustrates the normalized light emission (1 / 1O @ 428 nm) of the marker as described in Example 3. Figure 4 illustrates the normalized fluorescence emission (1 / 1O @ 427nm) of the marker as described in Example 4. Figure 5 illustrates the regularized glare emission (1 / 1O @ 540 nm) of the marker as described in Example 5. FIG. 6 illustrates the absorption spectrum of the label as described in Example 6. FIG. Figures 7-8 illustrate the absorption ratio (450 nm / 530 nm) of the marker as described in Example 6. Figure 9 illustrates the normalized fluorescence emission of the marker as described in Example 6 (1/1/10 at 550 nm). Figure 10 illustrates the fluorescence spectra of the marker as described in Example 6 in the absence of glucose and in the presence of 100 mg of molecular weight glucose. Figure Π illustrates the regularized fluorescence emission of the marker as described in Example 6 in the presence of glucose and lactate (1/1/10 at 550 nm). Figure 12 illustrates the regularized fluorescence emission (1/1/10 at 525 nm) of a marker exposed to glucose as described in Example 10. Figure 13 illustrates the normalized fluorescence emission (I / Ίο at 530 nm) of the lactate-labeled marker as described in Example 10. 85315 • 77- 200401034 Figure 14 shows the relative fluorescence emission (1 @ 430 nm) of the markers exposed to glucose and lactate as described in Example 11. Figure 15 shows the relative fluorescence emission (1 @ 43 () nm) of the markers exposed to glucose and lactate as described in Example I2. FIG. 16 illustrates the fluorescence of the target + hexate exposed to glucose and lactate as described in Example 13. FIG. 85315 78-

Claims (1)

'uU4〇l〇34, patent application park: 1 · A method for detecting glucose or its concentration in a sample that also contains alfa-hydroxy acid or beta-diketone, comprising: a) the sample Exposure to a compound having at least 2 glucose discriminating elements is oriented to make the interaction between the compound and glucose more stable than the interaction between the compound and alfa-hydroxy acid or beta-diketone, the compound is also Contains a detectable part with a detectable trait. When the chemical substance is exposed to glucose in the sample, the trait changes according to the concentration method; and b) any change in the trait is measured to determine The glucose or its concentration present in the sample, where the presence of alpha-hydroxy acid or beta-diketone does not substantially interfere with the assay. The method according to item 1 of Linggu Patent Park, wherein the compound has the following structure: I «4: ZW Rs1 -R- Rio r3 where -Ri is the same as R2 and 4 4 is not the same and is selected from the following: i) hydrogen; ii) the pKa of FP ^ and a hydrolytically stable substituent, 丨 丨 丨) Detectable part ^ iv) Linking group capable of attaching to a solid energy carrier or polymer matrix, the | body or matrix should include a detectable part as needed; -R3 hydrogen or (/ b @ An I 85315 group attached to a solid support or polymer matrix, the support or matrix including a detectable moiety as needed; -R4 is the same as or different from K · 5, and is selected from the following: 丨) hydrogen; Η ) Modified pKa and hydrolyzable stability of the h part, h〇 can detect the part or iv) a linker capable of attaching to a solid support or polymer matrix, the support or matrix including detectable if necessary Part;-each Z is independently carbon or nitrogen; -I is the same as or different from R7, and is i) a linking group having 0 or 10 adjacent or branched carbon and / or heteroatoms or capable of connecting to the solid state A linker attached to a support or polymer matrix, the support or matrix including a detectable moiety as needed; -R is selected from the following: i) aliphatic and / or Aromatic spacers, which include from 1 to 10 adjacent atoms selected from the group consisting of carbon, oxygen, nitrogen, sulfur and phosphorus; li) detectable moiety or Hi) capable of interacting with solid support or polymer A matrix-attached linking group, the support or matrix optionally including a detectable moiety;-each R8 is the same or different and is capable of interacting with a diol group present near glucose; and-9 and 111 () is the same or different, and is i} hydrogen; Η) can detect a part or lu) group, which is a) a linking group capable of attaching to a solid support or a polymer matrix. It needs to include a detectable portion, and / or b) include a functional group capable of changing the physical properties of the compound; /, the θ-labeled compound includes at least one detectable portion associated with it. β The method according to item 2 of the scope of the patent application, wherein R8 is selected from the group consisting of boron 85315 acid, acid ion, quinconic acid, phosphite ion, telluric acid, tellurate ion, germanic acid, germanate ion and A group of conjugates. The method according to item 3 of the patent application, wherein each R8 is a fluorenic acid group. 5. The method according to item 2 of the patent application, wherein the compound comprises at least two detectable moieties capable of being delivered to each other, and wherein the energy delivery is regulated by glucose present in the sample. 6. The method according to item 2 of the patent application park, wherein at least one of R, Ri, R2, R4, R5, R9, or R10 includes a fluorophore moiety, and further one of at least those groups includes The inhibitory moiety, and when the compound interacts with glucose in the sample, the fluorophore is inhibited or uninhibited. 7. The method according to item 2 of the scope of patent application, wherein the compound contains a fluorophore, and the interaction of the compound with glucose is used to regulate the luminescent of the fluorophore. 8. The method according to item 1 of the scope of patent application, wherein the sample is a physiological liquid. 9. The method according to item 8 of the application, wherein the physiological fluid system is selected from the group consisting of blood, plasma, serum, interstitial fluid, cerebrospinal fluid, urine, saliva, crystalline fluid, lymph, tears, sweat, and physiological A group of buffers. 10. The method according to item i of the patent application scope, wherein the compound is a sample exposed to a solution. 11. The method according to item 丨 of the scope of patent application, wherein the compound is immobilized on or in a solid support. 12. A method according to item 11 of the application, wherein the solid support system is a polymer matrix. 13. A method according to claim 1 in which the compound is associated with an implantable device, and wherein step a) occurs in vivo. 14. Method according to item 2 of the scope of patent application, wherein R is an anthracene residue; R1, R2, R3, R4 and R5 are hydrogen; r6 & R7 is a dimethylamine residue; each Rs is a boric acid group; One or both of R9 & R1Q is an aliphatic carboxylic acid residue; and each Z is a carbon. 15 * The method according to item 14 of the patent application, wherein one or both of 119 and 111 {) are propionic acid residues. 16. The method according to item 2 of the scope of patent application, wherein R is a hexamethylene residue; Ri, R2, R3, R4 and R5 are hydrogen; R6 and dimethylamine residues; each Rs is a borate group; 119 Is a dimethylformamide residue; R10 is a dimethylaminobenzyl residue; and each z is a carbon. 1 7. The method according to item 2 of the scope of patent application, wherein R is an anthracene residue; Rj, R2, r3, 114 and chi 5 are hydrogen; m7 is a dimethylamine residue; each Rs is a boric acid group; 119 Same as or different from 111 (), and is selected from the group consisting of methylpropanylsilylamino residue, methylpropoxyloxyethoxyalkyl residue, hydroxyethoxyalkyl residue and aminealkyl A group of residues; and each Z-series carbon. 1 8. The method according to item 2 of the scope of patent application, wherein the compound is selected from the group consisting of: 9- [N- (2-dihydroxyborylbenzyl) _N- [2- (2-methylpropionyloxyethoxy) Group) ethyl 85315, 200401034 amino] methyl] -10- [N- (2-dihydroxyborylbenzyl) _N_ [2 · (2-hydroxyethoxy) ethylamino] fluorenyl] anthracene; 9 , Η) -bis [N- (2-Dihydroxyborylbenzyl) · N_ [2 · (carboxyethyl) amino] methyl] Ε; 9,10-bis [N- (2-two-class Boryl benzyl) _Ν ♦ (Methylpropanylamino) propanylamino] methylanthracene; 9- [N- (2-Dihydroxyboryl group) -N- [3_ (methylpropyl Amido) propylamino] methyl] -10- [N- (2-dihydroxyborylbenzyl) #_ [2_ (2_hydroxyethoxy) ethylamino] methyl] anthracene; 9, 10-bis [^ (2-dihydroxyborylbenzyl) _ 仏 [2- (2-methylpropionyloxyethoxy) ethylamino] methyl] anthracene; 9,10-bis [N- ( 2-dihydroxyborylbenzyl) -N- [5-aminopentylamino] methyl] anthracene; and 9- [N- (2-diphenylborylfluorenyl) -N- [3- (methyl Propyl allylamino) propylamino] methyl] -10- [N- (2-dihydroxyborylbenzyl) -N- [6- (cyclohexanebenzylamino) hexylamino] methyl ] Anthracene; € 9- [Ν- (2- ) -N- [3- (Methylpropanylamino) propylamino] methyl] -10- [N- (2-dihydroxyborylbenzyl) -N- [2- (carboxyethyl) amine [Methyl] methyl] anthracene; 9- [N- (2-Dihydroxyborylfluorenyl) -N- [3- (methacrylamino) propylamino] methyl] -10- [Ν- (2 -Dihydroxyborylfluorenyl) -N- [3- (N-6- (9-E Carboxamido) hexylaminocarbonyl) ethylamino] methyl] anthracene; 9- [N- (2- Dihydroxyborylfluorenyl) -N- [3- (methylpropylamidino) propylamino] methyl] -10- [N- (2-Dihydroxyborylfluorenyl) -N- [6- (3-carboxypropylamidoamino) 85315 200401034 hexylamino] methyl] anthracene;, N- (3-methylpropylamidopropyl) -4- [2-N-[[2- (di Boryl) benzyl]-[6- (N- [2- (di-light boronyl) hexyl] -6-N- (3-carboxypropylamidoethyl) aminehexyl)] aminoethylamine Group] fluorene-1,8-dicarboxyfluorenimine; N-butyl-4- [2-N-[[2- (dihydroxyboryl) benzyl] _ [6 · (N- [2- (Dihydroxyboryl) fluorenyl] -6-N- (2-methylpropanamidoethyl) aminehexyl)] aminoethylamino] naphthalene-1,8-monochimonic acid and A group of its salts. 1 9. A compound with the following structure: -R! Is the same as or different from R2, and is selected from the following: 丨) hydrogen; ii) a substituent that changes the pKa of Rs and its hydrolytic stability, iii) a detectable moiety or iv) it can interact with a solid support Or a linking group attached to a polymer matrix, the exhaustion | the body or the matrix may include a manufacturable part as needed; ·. -R3 series chlorine or a linker capable of attaching to a solid support or a polymer matrix The matrix depending on f should include a dilutable moiety; R4l4R5 is the same or different, and is selected from the following: i) hydrogen; ii) a substituent that changes the pKa and hydrolytic stability of Rs, and iii) a detectable moiety or V ) A linking group to which a dog is attached to a solid support or a polymer matrix, and the support or matrix includes a manufacturable part as needed;-each "Z" is carbon or nitrogen; 85315 200401034 -R6 is the same as or different from R7, and is a linking group with 0 or 10 adjacent or branched carbon and / or heteroatoms or ii) a linking group capable of attaching to a solid support or polymer matrix, The support or matrix includes a detectable moiety as needed; -R is selected from the following: 丨) aliphatic and / or aromatic spacers, including from 1 to 10 selected from carbon, oxygen, nitrogen, sulfur and Adjacent atoms in a group of phosphorus; ii) a detectable moiety or ni) a linker capable of attaching to a solid support or polymer matrix, the support or matrix including a detectable moiety as needed;-each An Rs that is the same or different and that is protected as needed, when it is not protected, it can interact with a diol group present near glucose; and -9 is the same or different from R10, and System 丨) Hydrogen; 丨 丨) Detectable moiety or in) group 'It is a) a linking group capable of attaching to a solid support or polymer matrix, and the support or matrix includes a detectable moiety as needed, And / or b) include functional groups capable of modifying the physical properties of the compound; Includes detecting at least a part of its conclusion. 20. The compound according to item 19 of the scope of the patent application, wherein R8 is selected from the group consisting of boric acid, borate ion, arsenite, quinite ion, telluric acid, tellurate ion, germanic acid, and germanate ion (all Protected as needed) and combinations thereof. / * N 1. The compound according to item 20 of the patent application, wherein each r8 is a boronic acid group protected as necessary. 85315 ^ 2 > ^ 2 > 200401034 • The compound according to item 19 of the scope of application for a patent application, wherein the compound contains a fluorophore, and the fluorescence of the fluorophore is adjusted by the interaction of the compound with glucose. 23. The compound according to item 19 of the scope of patent application, wherein R is an anthracene residue; Rl, R2, r3, r4, and r5 are hydrogen; r6 and 117 are dimethylamine residues; each R8 is treated as needed Protected boric acid groups; one or both of r9 and r1g are aliphatic carboxylic acid residues; and each Z is carbon. 24. The compound according to item 23 of the application, wherein one or both of 119 and 111 () are propionic acid residues. 25. The compound according to item 1 of the scope of patent application, wherein R is an anthracene residue; R1, R2, R3, R4 and R5 are hydrogen; 116 and 117 are dimethylamine residues; each " A substituted boronic acid group is required; r9 is the same as or different from r1g 'and is selected from the group consisting of methacrylaminoalkyl residues, methacryloxyethoxyalkyl residues, and hydroxyethoxyalkyl residues A group of amine and amine alkyl residues; and each z is a carbon. 26. The compound according to item 19 of the scope of application for a patent, wherein the compound is selected from the group consisting of: 9- [N- [2- (5,5-dimethylboryl-2-yl) fluorenyl] -N_ [2- (2-Methylpropionyloxyethoxy) ethylamino] methyl] -lO- [N- [2- (5,5-dimethylborofluoren-2-yl) yl] -N -[2- (2-hydroxyethoxy) ethylamino] methyl] anthracene; 9- [N- (2-bis # iborylbenzyl) -N- [2- (2-fluorenylpropionyloxy) Ethoxy) ethylamino] methyl] -10- [N_ (2-dihydroxyborylbenzyl) _N_ [2- (2-hydroxyethoxy) ethylamino] methyl] anthracene; 9, 10-bis [N- (2-dihydroxyborylfluorenyl) -N_ [2- (carboxyethyl) amino] methyl] 85315 200401034 anthracene; 9,10-bis [N- [2- (5, 5-Dimethyl ππ-2-yl) benzyl] _ν_ [3 · (methylpropylamido) propylamino] fluorenylanthracene; 9,10-bis [N- (2-dihydroxyboron) Sulfenyl) -N- [3- (methylpropanylamino) propylamino] methylanthracene;-9- [N- [2- (5,5-Dimethylboron) ) Yenyl] -N- [3- (methylpropylalanino) propylamino] methyl] -1 0- [N- [2- (5,5 -Dimethyl lan ?? _ 2_yl ) Nyl] -N- [2- (2-hydroxyethoxy) ethylamino] methyl] anthracene; φ 9- [1 ^-(2-bis # fluorenylbenzyl)-] ^-[ 3- ( Methylpropionate amine) propylamine] methyl] -10- [N- (2-Dioxoborohydride) -N- [2- (2-Ethoxy) ethylamino] fluorenyl ] Anthracene; 9.10-bis [N- [2- (5,5-dimethylboron ρ-2-yl) methyl] -v- [2- (2-methylpropanyloxyethoxy) ethene Amine] methyl] anthracene; 9,10-bis [1 ^-(2-diborobenzylbenzyl)-> 1- [2- (2-methylpropanyloxyethoxy) ethylamine [Methyl] methyl] anthracene; 9.10-bis [N- (2-dialylbenzyl) -N- [5-aminopentylamino] methyl] -anthracene;-· 9- [N- (2 -Distilbenzyl) -N- [3- (methylpropylamido) propylamino] methyl] -10- [N- (2-dihydroxyborylbenzyl) -N- [6 -(Cyclohexanecarboxamido) hexylamino] methyl] anthracene; 9- [N- [2- (4,4,5,5-tetramethyl-1,3,2-dioxorane ) Fluorenyl] _ν- [3- (methylpropylamido) propylamino] methyl] -10- [N- [2- (4,4,5,5-tetramethyl-1,3, 2-dioxorimido) fluorenyl] -N- [6- (cyclohexanecarboxyfluorenylamino) hexylamino] methyl] anthracene; 85315 200401034 9- [N- (2-dioxophenylyl) ) -N- [3- (methylpropylamido) propylamino] methyl] -H)-[N- (2-dihydroxyborylhenyl) -N- [2- (carboxyethyl) amine Methyl] methyl ] Anthracene; 9- [N- [2- (4,4,5,5-tetramethyl-1,3,2-dioxo group)] _ n_ [3_ (methacrylamino) Propylamino] methyl] -10- [1 [2- (4,4,5,5-tetramethyl-1,3,2-dioxofluorenyl) methyl] -N- [2- (chito (Yl) amino] methyl] anthracene; 9- [N- (2-Dihydroxyborylfluorenyl) -N- [3- (methylpropylamido) propylamino] methyl] -10- [ N- (2-Dihydroxybenzyl) -N- [3- (N-6- (9-E Carboxamido) hexylaminocarbonyl) ethylamino] methyl] E; 9- [N -[2- (4,4,5,5-tetramethyl-1,3,2-dioxorimido) benzyl] _N_ [3 · ((Methylpropanamine) propylamino) methyl] -10- [Ν- [2- (4,4,5,5-tetramethyl_1,3,2-dioxo taste) Ethyl] -N- [3- (Ν-6- (9- Onion carboxyfluorenylamino) hexylaminocarbonylethylamino) methyl] anthracene; 9- [N · [2- (4,4,5,5-tetramethyl-1,3,2-dioxorane) ) Ethyl] · ν_ [3- (methylpropylalanino) propylamino] methyl] -1O- [N- [2- (4,4,5,5-tetramethyl-1,3, 2-dioxoboryl) benzyl] -N- [6- (3-carboxypropanamido) hexylamino] methyl] anthracene; 9- [N- (2-dihydroxyborylbenzyl)- N- [3- (methylpropylamido) propylamino] fluorenyl] -10- [N- (2-dihydroxyborylbenzyl) -N- [6- (3-carboxy Fluorenylamino) hexylamino] methyl] anthracene; Ν- (3 · methylpropylamidopropylpropyl) _4- [2-N-[[2- (dihydroxyboryl) fluorenyl]-[ 6- (N- [2- (Dihydroxyboryl) fluorenyl] -6-N- (3-carboxypropylamidoethyl) aminehexyl)] aminoethylamino] fluorene-1,8- Dicarboxydonkey imine; Ν-butyl-4- [2-N-[[2- (dihydroxyboryl) fluorenyl] _ [6- (Ν- [2- (dihydroxyboron • 10- κ ^ 3 85315 200401034 Group) Ethyl] -6-N- (2-methylpropanamidoethyl) aminehexyl)] aminoethylamino] Tetra-1,8-dicarboximimine and its A group of salts. 27. A detection system for detecting glucose or its concentration in a sample that also contains alfa-hydroxy acid or beta-diketone, comprising a compound having the following structure: r3 rio -R 丨 is the same as or different from R2, and is selected from the following: i) hydrogen; u) a substituent that changes the PKa and hydrolysis stability of the R8 moiety, ii) a detectable moiety or IV) that can interact with the solid state A linker attached to a support or polymer matrix, the support or matrix including a detectable portion if necessary;-Rs is hydrogen or a linker capable of attaching to a solid support or polymer matrix; Need to include the detectable part; -R4 and Rs are the same or different, and are selected from the following: i) hydrogen; 丨 丨) the PK & part of the change & Or iv) a linker capable of attaching to a solid support or matrix of a cold compound, the support or matrix including a detectable portion as needed; ^ S-each Z is independently hard or nitrogen; ice 6 and R7 is the same or different, and is i) a linking group with 0 or 10 contiguous or chain completion and / or heteroatoms or ii) _ with a solid support or? The character base is attached and connected to the base. The support or matrix includes a detectable portion of the detectable 85315 -11- 200401034; -R is selected from the following: i) aliphatic and / or aromatic spacers, including 1 to 10 adjacent atoms selected from the group consisting of carbon, oxygen, nitrogen, sulfur, and phosphorus; ii) a detectable moiety or iii) a linking group capable of attaching to a solid support or a polymer matrix, the support The body or matrix includes detectable moieties as needed;-each of which is the same or different and is protected as needed, when it is unprotected; it is capable of interacting with glycol groups present near glucose ; And -R9 is the same as or different from R10, and is i) hydrogen; ii) a detectable part or iii) a radical 'its a) a linking group capable of attaching to a solid support or a polymer matrix; the support; Or the matrix may include a detectable portion as needed, and / or b) include a functional group capable of modifying the physical properties of the compound; however, the book-based marker compound includes at least one detectable portion associated with it. 28. The detection system according to item 27 of the scope of patent application, wherein the ruler 8 is selected from the group consisting of boric acid, borate ion, quinconic acid, arsenite ion, telluric acid, tellurate ion, germanic acid, and germanate A group of ions (all protected as needed) and their conjugates. 29. The detection system according to item 28 of the scope of patent application, wherein each ruler 8 is a boric acid group protected as required. 30. The detection system according to item 27 of the scope of patent application, wherein the compound contains a labor group, and the fluorescence of the fluorophore is adjusted by the interaction of the compound with glucose. 85315 200401034 L The detection system according to item 27 of the patent application park, where R is anthracene residue; Ri, R2, R3, R4 and R5 are hydrogen; R6 and R7 are dimethylamine residues; each R8 is Protected boronic acid groups as required; one or both of R9 & R1 () are aliphatic carboxylic acid residues; and each Z is carbon. 32. The detection system according to item 31 of the scope of patent application, wherein one or both of ruler 9 and 1110 are propionic acid residues. 33. The detection system according to item 27 of the scope of patent application, wherein R is anthracene residue; Ri, R2, R3, R4 and R5 are hydrogen; 116 and 117 are dimethylamine residues; each R8 is as required Substituted boronic acid group; 119 is the same as or different from R1Q, and is selected from the group consisting of methacrylaminoalkyl residues, methacryloxyethoxyalkyl residues, and hydroxyethoxyalkyl residues And amine alkyl residues; and each Z-based carbon. 34. The detection system according to item 27 of the scope of patent application, wherein the compound is selected from the group consisting of: 9- [N · [2- (5,5-dimethylborofluoren-2-yl) benzyl] -N- [ 2- (2-methylpropanyloxyethoxy) ethylamino] fluorenyl] -10- [N- [2- (5,5-dimethylborofluoren-2-yl) benzyl] -N- [2- (2-hydroxyethoxy) ethylamino] methyl] anthracene; 9- [N- (2-dihydroxyborylbenzyl) -N- [2- (2-methylpropane Ethoxyethoxy) ethylamino] methyl] -10- [N- (2-dihydroxyborylfluorenyl) -N · [2- (2-hydroxyethoxy) ethylamino] methyl ] Anthracene; 9,10-bis [] ^-(2-Dihydroxyborylbenzyl)-> 1- [2- (carboxyethyl) amino] methyl; | Anthracene; 9,10-bis [ Ν- [2- (5,5-dimethylborofluoren-2-yl) fluorenyl] -N- [3- (fluorenylpropylamido) propylamino] methylanthracene; 85315 • 13- 200401034 9,10-bis [N- (2-dihydroxyborylbenzyl) _n- [3- (methacrylamino) propylamino] methylanthracene; 9- [N- [2- (5,5 -Dimethylboryl-2-benzyl) benzyl] -N- [3- (fluorenylpropylalanino) propylamino] methyl] -10- [N- [2- (5,5- Dimethylborofluoren-2-yl) benzyl] -N- [2- (2-hydroxyethoxy) ethylamino] methyl] anthracene; " 9- [N- (2-Dioxonyl Benzyl)- N- [3- (Methacrylamino) propylamino] methyl] -10- [N- (2-Dihydroxyborylbenzyl) _N_ [2- (2-hydroxyethoxy) ethylamino ] Methyl] cis; · 9.10-bis [N- [2- (5,5-dimethylborofluoren-2-yl) benzyl] -N- [2- (2-methylpropanyloxyethyl) (Oxy) ethylamino] methyl] anthracene; 9,10-bis [1 ^-(2-dihydroxyborylbenzyl)-: ^-[2- (2-fluorenylpropionyloxyethoxy) ) Ethylamino] methyl] anthracene; 9.10-bis [N- (2-dihydroxyborylbenzyl) -N- [5-aminopentylamino] methyl] anthracene; and 9- [N- ( 2-dihydroxyborylbenzyl) -N- [3- (methacrylfluorenylamino) propylamino] i methyl] -10- [N- (2-dihydroxyborylfluorenyl) -N- [ 6- (Cyclohexanecarboxamido) _hexylamino] methyl] anthracene; 9- [1 ^-[2- (4,4,5,5-tetramethyl-1,3,2-di Oxyboryl) benzyl]-: ^-[3- (methacrylamino) propylamine] methyl] -10- [1 ^-[2- (4,4,5,5-tetramethyl -1,3,2-dioxorimido) benzyl] -N- [6- (cyclohexanecarboxamido) hexylamino] methyl m; 9- [N- (2-dihydroxypentyl Benzyl) -N- [3- (methylpropylamido) propylamino] methyl] -10- [N- (2-dihydroxylamino) -N- [2- (carboxyethyl ) Amino] methyl] anthracene; 85315 -14- 200401034 9- [N- [2- (4,4,5,5-tetramethyl-1,3,2-dioxorimido) benzyl] _n- [3- (methylpropylamidinyl) propylamine Yl] methyl] -lO- [N- [2- (4,4,5,5-tetramethyl-1,3,2-dioxofluorenyl) benzyl] -N- [2- ( Ethyl) amino] methyl] anthracene; 9- [N- (2-Dioborylfluorenyl) -N- [3- (methacrylamino) propylamino] methyl] -10- [ N- (2-Dihydroxyborylbenzyl) -N- [3- (N-6- (9-anthracenecarboxamide · -yl) hexylaminocarbonyl) ethylamino] methyl] anthracene;-9 -[N- [2- (4,4,5,5-tetramethyl-1,3,2-dioxyboron) benzyl] · ν- [3- (methylpropylamido) propylamine Group] methyl] -10- [1 ^-[2- (4,4,5,5-tetrafluorenyl-alkyl 1,3,2-dioxolidine) benzyl] -N- [3- ( Ν-6- (9-anthracene-pyridinylamino) hexylaminocarbonylethylaminomethyl) anthracene; 9- [N- [2- (4,4,5,5-tetramethyl-1,3, 2-Dioxoboryl) benzyl] _N- [3- (methacrylamino) propylamino] methyl] -1 (HN- [2- (4,4,5,5-tetramethyl_ l, 3,2-dioxoamido) N] [N- [6- (3-Episylpropylamido) hexylamino] methyl] anthracene; 9- [N- (2-Dioxoboryl Benzyl) -N- [3- (methylpropylalanino) propylamino] methyl] -10- [N- (2-dihydroxyborylbenzyl) _Ν_ [6- (3 · Propylpropylamido) hexylamino] methyl] anthracene; N- (3-methacrylamidoaminopropyl) _4_ [2_ν _ [[2_ (dihydroxyboryl) benzyl] _. [6- (N- [2- (Dihydroxyboryl) fluorenyl] _6_ν_ (3-carboxypropylamidoaminoethyl) aminohexyl)] aminoethylamino] fluorene_1,8-* Ν-butyl- 4- [2-N-[[2- (Dihydroxyboryl) benzyl] _ [6_ (Ν_ [2 · (Dihydroxyboryl) fluorenyl] -6-N- (2-methacrylamide Group consisting of aminoethyl) aminohexyl is amineethylamino] nelidene-1,8-dicarboxyamidoimide and its salts. 85315 -15-