JPS5857356A - Anthroyl nitrile, its preparation and fluorescent reagent - Google Patents

Abstract

NEW MATERIAL:A compound of formulaI(any one of X<1>, X<2> and X<3> is carbonyl nitrile; the rest are H). EXAMPLE:9-Anthroyl nitrile. USE:An active component in analytical reagents, e.g. fluorescent reagents, and particularly useful as detecting and determining various steroids which are minor components in the living body. PROCESS:An anthroyl halide of formula II[any one of Y<1>, Y<2> and Y<3> is carbonyl halide (COX: X is halogen); the rest are H]is reacted with a trialkylsilyl cyanide in an anhydrous organic solvent, e.g. methylene chloride or chloroform, to give the corresponding compound of formulaI. In the process, a metallic iodide, e.g. zinc iodide, as a catalyst is added if necessary to the reaction system to give the aimed compound in high purity and yield. In using the compound of formulaIas an active component for the fluorescent reagent, the active component is preferably dissolved in an aprotic organic solvent, e.g. chloroform or tetrahydrofuran (THF), and an organic base, e.g. triethylamine, is present in the solution.

Description

[Detailed description of the invention] A relates to anthroylnitrile, its production method, and a fluorescent reagent However, compounds containing A/pool hydroxyl groups are produced using high performance liquid lithography (hereinafter referred to as HPL).
C) can be detected with high sensitivity! I is very strong, and I try to convert it into a fluorescent substance from a strong ultraviolet absorbing substance.
However, to the best of our knowledge, a nine-gravelling agent with excellent reactivity, sensitivity, and stability has not been found.

This invention was made to solve this interlayer problem. 94 ◇ That is, according to this development MK, a compound having a β-bonylnitrile (-cock) group in the anthracene ring reacts with a 7μ chol hydroxyl group, which has low reactivity. It was found that it is excellent as a fluorescent reagent in various aspects.

Thus, according to Jin's invention, the general formula (■): ][1xl [wherein, one of -XI, X' and xa is the force μ
A compound represented by the following formula is provided: poeN nitol (-coax) group (To) and others representing hydrogen atoms. The specific compound salts are: 1. or 9-antogastricylnidol. All of these compounds are metal compounds that have not been described in any literature.

The metal compound of the general formula (1) belongs to the acyl nitrile group, and is generally carried out by the i method in which the corresponding halide is reacted with cupric cyanide, sodium cyanide [to, x, O.mlg&W, P,
Weber, Tetrahedron L@tt.
g @! 175 (1174))
However, the compound of general formula (1) can be prepared by converting the corresponding ^lide in an anhydrous organic solvent! li! It has been found that when a metal iodide is added as a catalyst to KI6 and the mixture is reacted with totsuakisiliμ cyanite, high purity and high yield can be obtained.

Thus, according to one aspect of the present invention, the corresponding general formula (I): (a) group that represents an atom, and the others represent a hydrogen atom. ]Ant stomach I μ halide is anhydrous 1! A method for producing a compound of general formula (1) is provided, which comprises reacting it with tri-μ-silyl p-cyate in medium #1.

Using this method, the product is obtained in high purity as described above and is suitable for use as an analytical reagent.

The compound of general formula 1) used in the above method can be obtained, for example, by introducing a γp-koxy*py-poxfit group into the corresponding anthot-tetraylbutyl group by Grignard reaction, and hydrolyzing it with an alkali metal acoat. As the 0-doria-kylsilylcyanite which can be obtained by processing with a chemical compound such as oxalyl chloride, it is preferable to use tofumethysilylcyanite or triethylsilylcyanite. In addition, as a reaction solvent, ordinary organic solvents (for example, methylene chloride, chloroform, dimethylene chloride, chloroform, dimethyl chloride, tetrahydrocerane, etc.) in reactions using silyl north side can be used. Of course, this reaction is preferably carried out in an anhydrous state by adding a metal iodide (for example, zinc iodide) as a catalyst.

In addition, specific examples of the amount of each reactant to be used, reaction time, reaction temperature, etc. are explained by the following synthesis examples. However, these various conditions may be changed as appropriate.

The compound of general formula (1) thus obtained can be used as a fluorescent reagent.

Thus, one aspect of the invention is to provide a fluorescent reagent comprising a compound of general formula (I) as an active ingredient.

This active ingredient Fi can be used alone, but
Preferably, the reagent is dissolved in an aprotic organic solvent. In particular, when the compound of general formula +13 is used as a fluorescent reagent, the presence of an organic base brings about good results in the reaction with the test sample, so it is preferable to add an organic base to the reagent.〇Aprotic Specific examples of the organic solvent include lysate roform, acetonitrile, benzene, acetate ethane, dihydrofurin, acetone, and the like.

Among these, acetonitrile is the best. Examples of the organic base include tertiary alkyl anions such as triethyl 7one, tomethane, and quinuclidine.

The ratio of the aprotic organic solvent to the organic base is usually 1 to 1 (approximately 1:0.1).In addition, the concentration of the compound in the general formula in these mixtures is particularly Examples include, but are not limited to, Q, 1 to 0.6 weight.

The fluorescent reagent according to this invention has a primary alcoholic hydroxyl group,! Class A/: Reacts specifically with y-* hydroxyl groups and pheno-μ hydroxyl groups. Kudashi 2 with steric hindrance
Among the class hydroxyl groups (for example, the 11β-position, the 17-position, and the axial hydroxyl group of steroids), the tertiary 7-chole hydroxyl group and the wipe hardly react with each other.

Especially among the compounds of this invention! -When using anthocytoethomal, it has the highest reactivity and a high esterification rate with a hydroxyl group can be obtained in a short time, and is most preferable as a fluorescent reagent.Specific examples of these will be explained in the examples below.

It is desirable to use an excess amount of the fluorescent reagent with respect to the test sample to sufficiently cause the umbrella reaction. It has been found that this method has no effect on the fluorescence analysis of the sample being tested.

When measuring actual dust, it is first reacted with a test sample and then subjected to high-impermeance liquid chromatography.

The fluorescence of the nine esters formed can be detected using a fluorophotometer.

In addition, when steroids are used as the test sample, the detection sensitivity is w@pt (co~tezo μ) in a normal phase system with a high liquid shielding liquid, and at anti-pI (9 toco-p acid) in a reverse phase system. Ata is better than the conventional one. First, the fluorescence wavelength is 4 nm (maximum excitation wavelength 37 nu) when using 1-anthronitrile, and 46 am (maximum excitation wavelength 37 nu) when using 9-T nitride. The maximum excitation wavelength is 36921 (Otori).

Thus, the fluorescent reagent according to the present invention is particularly useful for detecting and quantifying various steroids that are trace components in living organisms.

Synthesis Example 1 9-anthoylnidoturu: T7trace/-9-carbo/acid (ry) t-m mixed and stirred in hydrated methylene chloride (is1m/), and oxalyl chloride (2-) was added thereto, After heating under reflux for 1 hour, the solution was concentrated under reduced pressure. The residual liquid was diluted with anhydrous methylene chloride (15i
1) Dissolve K, add zinc iodide and tomethylsilyl cyanide (1!11 t) from the catalyst, and stir at room temperature for 2 hours. After concentrating the reaction solution under reduced pressure.

The residue was recrystallized from a mixture of n-hexane/methylene chloride to obtain the title compound (800) as orange needles.

The physical properties of the obtained chemotactic compound are as follows.

Melting point: 143-144°C IR (Rion Ho vA) rta! Lx: tz<o(a=
N> and 167G (0=O) cs+-' M811/m: 231 (M”) and !O6 (CM-CN
)") Elemental analysis: Actual value Calculated value (0,, H,
No) 0 8g! 1 811.10H4,
938,9ffi M 1! iJ3 6.06 synthesis N! The same procedure as in Synthesis Example 1 was carried out, except that l-antogastricylnide-sugaruanthracene-9-ponic acid was replaced with anthralin-1-ponic acid,
The title compound (860 Da) was obtained as orange needles.

The physical properties of the obtained chemotactic compound are as follows: 2i! Ri de Atsuku.

Melting point: 164-165℃ IIL (RIaaho*A) 1111X: 224@
(OmN) and 1675 (C-0) cm'' M811/K: s3t (M+) and addition S ((M-
(3N) +) Elemental analysis 2 Actual value Calculated value (01
,11,ND)0 8847 81.11
H8,71892 N 5.96 6.66 Analysis example As a test sample, each Sudetei-id (1 #g
), and each of these was dissolved in an acetonitrile solution containing trietherazine (0.1 wt.). Anthroylnitrile or! was prepared according to the synthesis example above. -Ant fog i β nido 9le t e @ s
1 and reacted at 6°C, and measured the Nisder conversion rate until completion of the esterification reaction or after 1 hour.Measurements were made by subjecting the resulting esterum to true speed liquid chromatography. This was done by separating and measuring the fluorescence wavelength of 460 nm or 476 nm.

The nine high-speed liquid pomatograph devices used and the measurement conditions are as follows.

Equipment: Waters 6...M (USA, Waters
Association) Detector: 650-10LCg photometer (newly manufactured by Hitachi) Katsumu: μ Borashi * (1 fee) X1/4
inch; mobile phase, L3 typhosphate buffer/ethanol-A/) The measurement results were as shown in Table 10.

In addition, 6 in the table is equatorial, qe is quasi catal, & is axial, and prba is first class.

phen represents a phenolic hydroxyl group. Also, the * mark is the result when quinuclidine was used instead of triethylamine.

As shown in Table 1, in 9-anthroylnitrile and l-
Both antholylnitrile (uniformly, it was found that the reaction with a violet alcoholic hydroxyl group was completed in a short time) and IB) hardly reacted with a sterically hindered secondary alcoholic hydroxyl group. It can be seen that (5) hardly reacts with the equatorial hydroxyl group, but reacts with a high efficiency with the equatorial hydroxyl group. It has also been found that the equatorial hydroxyl group at the 3c position of cholic acid exhibits excellent reactivity when quinuclidine is used as a base. In addition,
Both (a) and (b) had good reactivity with the phenolic hydroxyl group.

From the above results, it was found that the hydroxyl group at a specific position was selectively esterified, so that it could be placed in place.

Claims (1)

[Claims] (In the formula, Xi,
/& 蟲βnidvru(-coam) group) and others indicate hydrogen atoms. ] Shown in 4 Jkl Eto! Le. L 1-Antoy A/&Tosugarumata I! Claim 11 [1Ij11
tnt■i〃nttψk mentioned in the section. [In the formula, Xi, 1M, lhi
-)illpbonylnitrile (-000!N) group, and the others represent hydrogen atoms. ] A fluorescent reagent containing anthruylnitrile as an active ingredient. t Active ingredients! -Antofoil nitrile or 9-a/
4. The fluorescent reagent according to claim 3, which is Ryl. & Claim 3 or! where the active ingredient is dissolved in a non-boutonic organic solvent! Fluorescent reagent according to item 4.叡 The fluorescent reagent according to any of claims S-S, wherein the active ingredient is understood to be a non-boutonous organic solvent and an organic base. L The non-bouton organic solvent is acetonitrile, and the organic salt iM is tosyetean. Claim 4'y
Fluorescent reagent described in LK. General formula (I): Yl 73 [In the formula, any one of 71 , 71 and ya is a puehatsuide (-, -COX; in the formula, Xij represents a -rodane atom) group] Other minerals Indicates a hydrogen atom. The corresponding general formula (1) is obtained by reacting an)Dyl I-phyto represented by the above with triacylsilicyanite in an anhydrous organic solvent.
: % formula % (in the formula 3 (1, F or Hm, the contact force is p!
In the μnidocyl (-COO)IT) group, p and the others represent hydrogen atoms. ] To obtain anthroylnitrile shown in 1) The method for producing anthroylnidol ψ. East: The manufacturing method according to claim 8, which is carried out by adding a metal iodide as a catalyst. 14L) ψA ~ Kirsiri〃Cyanite is trimethip
9. A method for producing a compound of claim 8 and claim 9, which is cyanite.