JPS5944352A - Water-soluble diaminodiphenylmethane derivative and its use - Google Patents

Abstract

NEW MATERIAL:The compound of formula I (R<1> and R<3> are H, lower alkyl, 2-3C sulfoalkyl, etc.; R<2> and R<4> are 2-3C sulfoalkyl or hydroxysulfoalkyl; R<5> and R<6> are H, methoxy, hydroxy, or lower alkyl) and its salt. EXAMPLE:Di-{4-[N-ethyl-N-(2-sulfoethyl)amino]phenyl}methane disodium salt. USE:A pigment-forming substance. It has high solubility in buffer solutions having various pH values, and a reagent containing the compound can be prepared easily without adding a surface active agent and an organic solvent thereto. Since the wavelength for the absorption peak of the condensation product with 4- aminoantipyrine is mostly >=560nm, the influence with bilirubin and hemolysis can be avoided. PROCESS:The compound of formula I can be prepared by reacting the compounds of formula II and formula III with formaldehyde.

Description

DETAILED DESCRIPTION OF THE INVENTION Water-soluble + 1"di"rminodif, a nilmethane derivative and 1,000 salts (1-F [abbreviated as the compound of the present invention-1)] and the present invention [7] The composition comprises: iM oxidant,;
).

All of the above compounds of the present invention are found in the Fi' standard and useful substance at the end of the literature.
Pigment-forming substance and J-C when killing hydrogen peroxide in coexistence with 7 case or a similar oil-mediated substance.
Useful.

(Clinical assays using enzyme tests are highly selective analytical methods due to the substrate specificity of enzymes, and are complex)
It has become rapidly popular in recent years because it allows various in vivo components to be easily injected into the body without being affected by +7. For example, glucose, flic acid, cholesterol, etc. in body fluids are processed by glucose oxidase, uricase,
It is broken down by the enzyme cholesterol oxidase and produces the corresponding 115 hydrogen gas. Therefore, various in-vivo components can be measured by quantifying this hydrogen peroxide. Conventionally, the method of quantifying hydrogen peroxide produced by such a mechanism is often based on oxidative condensation coloring, and for example, the following pigment-forming substances have been reported.
-dimethylaniline; toluidine; N,N-diethyl-
m-toluidine, etc.) (2) Phenylenediamine derivatives and diamines such as benzidine and ginseng (3) Phenol derivatives and naphthol derivatives (
4) Leucotriphenylmethanes (leucomalachite green and leucophenolphthalein) +51
2.6-cyclophenol indolph, leucotriphenylmethanes and 2.
6-cyclophenol indophenol and ('1
')'> Mushi K fjL use Saneru, but aniline slander conductor,
4 for phenylene diamine, high conductor, and phenol residue conductor.
-Aminoandibirine, 3-Mediru 2-pen 7'1,000 azolinone hydrazone (1!'F 1vBTnJ and abbreviation +
), 9j is used together with a color former of 1'5 and 1.

Appropriate amount to act on, 1llll
(1) It differs depending on the type of enzyme that oxidizes and decomposes the compound.
, Runo, Terorun] in one year, j p IF
6.5-80.

Oj-breast pH 5~q(")帥pHにり)" Cloth [, 7. However, the color of the above-mentioned pupa is formed, especially monoamine'(ri,;-Zmin) under the conditions of D115~9. In this case, it is unfortunate that Mt is dissolved in water. (7 or l)
11. The coexistence of surfactants and organic solvents will deactivate the fermentation. 48-1 These publicly known colors 2 carvings 117-1'! 11σ) 4, Stock solution (・1・1゛Liquid γ constant 1′) is poor, and 1〃( is held up while detecting concentration changes over time.

Tree S) Surrounders use 4-nominoampvirin, police, l
As a result of extensive research, I used a pigment-forming substance that undergoes oxidative condensation with B T 11 and has good water solubility as much as possible in M5, and as a result of the above-mentioned Mr. Sha (1), 7r- SaJl
, I found a compound in Group 33 of I-jroft-, the compound of the present invention, and %4 RT) I, and performed oxidative condensation with Vermexiter-V; Quantitatively form a blue dye represented by the formula (n) (.
;2 and hair ξL and redundant 11 (−, 1>^ (1) (MBTI() (b) ′.“”・ By the way, ?E and j f are dedicated to f, no A
and Ti, tl-deca dimethylaniline, and 4-aminoan-1-biline J
-'s 1+i% synthetic compound has a similar absorption wavelength of 500 t-, 2D nm, which is close to 4-1; Since the UV absorption ranges over a wide range from the ultraviolet 9 region to around 500 nm, there is a possibility that a slight positive error may occur.1.The compound of the present invention represented by the general formula (1) ir: Maximum 1 of the synthesis product with F, MBTH
Since the 1"L absorption wavelength is mostly 560 ++ m or more"5, it is possible to avoid the effects of bilirubin and hemolysis7).
There are significant advantages.

In addition, when the inventive compound and MBTHipH7 are mixed and dissolved in the W surgical solution (2), when beroogysodase and hydrogen peroxide are added, the molar extinction coefficient (r, mo) at the maximum absorption wavelength is
(・m"' l is 5.1) x ] 0' not 1, 6
．． The color of OX 1.0' is 1]. This was measured under the same conditions using full phenol and 4-aminoantipyrine. 1), the molar extinction coefficient at the absorption wavelength is approximately 5. OX 1. (It's only about a bit.

Therefore, the determination of perr1 hydrogen chloride using the compound of the present invention
Method Ij can be said to be a very sensitive method.

The compounds of the present invention have good solubility in laxative solutions of various pH values, and there is also the advantage that there is no need to add surfactants or organic solvents, and reagent preparation is simple.

Now, the first-order general formula (1
1) and (II') P'R" (Ill) (11'1(λ, medium u
I, R1, P, 4, Rfi, and R* have the same meanings as in the above general formula (1)) It can be synthesized by reacting formalin. Specific examples of the compound of the present invention represented by the general formula (1) synthesized in this way are shown in Table 1, and the glaze point and pH 7 of the compound are shown in Table 1. Table 2 shows the maximum absorption wavelength max (nm) of the oxidative condensation dye.

1 ″ (Ma (3□ 1 ... ＆1 11q U
1) 1→
Group b -〉 Ta
0-P I○ "trI" In +Q Z ・; Kawada (7 p4 ()' Now, when carrying out the quantitative determination according to the present invention (・11) The pigment-forming substance of the present invention (a) Peroxidation effect A certain substance and (b) a color forming agent are used. Examples of the former (i) include peroxidase, as well as transition metals such as iron ions, and examples of the latter (b) include In addition to MBTH, examples include 4-aminoantipyrine.However, these are just examples and the present invention is not limited thereto. The manufacturing method (Examples 1 to 6) and the fixed-basis method using the same (Example 7) will be described, but the present invention is not limited to the following examples unless the gist thereof is exceeded.

Example 1 Preparation of compound FA) 103 g of N-ethyl-N-(2-sulfonidel)aniline sodium salt was dissolved in 20 ml of water, and 37% FA/?
71.78ml of water, add 0.20ml of 90% formic acid and 35
N-methylaniline was added while maintaining the temperature at °C to carry out the reaction. The obtained crystals were collected and recrystallized with methanol to give white di-14-(N-ethylAt-N-(2-sulfoethyl)amite phenylymethane disodium salt8.25
Obtain El; Yield: 77.8 TLC (Nrica gel, 0
2N ammonia water saturation ■]-butanol) Rf=0.2
5′H-NMR((L,S-nMsO)J(T
M R)j+111n 1.12 (t;, ,T==7.8T(,6JT
1 2.44-2.79 (m.

FIT() 2. R3-3,41 (m, 4H)
364 (s, 2H) 6.32 ((+, J=8
．． 2H11,,4H) 6.74 ((1, J=8.
2H, 4H) IR (“-”) 1608 1.263 12
+10 Example 2- Preparation of compound (B) Dissolve 489 g of N-(3-sulfopropyl)aniline sodium salt in 10 ml of water and maintain at 35° CIc 37
% formalin 0, F19m/, 90% formic acid 0.10 m
+! Add and react. The obtained white crystals are designated as P and f
fY and recrystallized from an acetone-water mixed solvent.

4.23 g of di(4-(N-(3-sulfopropyl)aminocophenyl)methane 2-sodium salt is prepared. Yield: 844% ) Rf-0,25 1,6(1-2,16(m, 4H) 2.38-2
．． 71i (m, 4H) 302 (Jin, ,T
=6.2H, 4H) 3. ! '+8 (J
21-1) 6.47 (d, J=8.2) (,4H)
6.90 (d,, J=8.2H, 4H) Engineering R (a++-') 3320 1610
12831169 1050 Example 3 Preparation of compound (D) N-ethyl-N-(3-sulfopropyl)-γ;, dissolved 5.47 g of phosphorus sodium salt in water LOmlK, 37%
, j; 7L Mari: y O, 89pi, 9o% formic acid 0.
Add 1 (1 m/!) and keep 35' OK. 15 (U) Add N-methylaniline to carry out the reaction. Take the obtained white crystals and recrystallize with a THF-methanol mixed solvent to obtain di(
Obtain 4.95 g of 4-(N-ethy#-N-(3-sulfopropyl)amine)fuyo,nil"methane disodium salt. Yield 883% T1+af silica gel, 0.2N anthonia water saturated n-butano -Rf=(130 1H-NMR(d6-DMSO)Jppm(TMS)1
1. 10 ft, J = 7.7H, 6H)
1.68-2.18 (m.

4H) 2.40-2.75 (m, 8'H
) 2.81-3.30 (m, 4H) 3
．． 52 (8,2H) 6.43 (c+,,
J=8.2. H, 4H) 6. F18 (d
, J=8.2H,4H)IR(cm-')
3480 1602 12611 1 78
1, 041 4 examples 4 Compound (Production of Gl) 3-Hydroxy-N-ethyl-N-(3-sulfopropyl)aniline sodium + 4.tll, 50 g in water 20
m1K (i, 37 formalin 1.8 Q m+°
, 90 Tigi rP0.2 (l ml' f plus 35'
Add and react N-methylaniline while keeping the Cvc. The resulting precipitate was crushed and crystallized while decanting several times with acetone, and the resulting bluish-white crystals were collected.
．． Recrystallize with methanol.

White di(2-human 1) paper 4-[N-ethyl-N
-(3-sulfopropyl)aminocophenyl)methane 2
Sodium salt 6,') 7B is 〒+A. Yield: 53.0 t TL (! (Silica gel, (1,2), 1 t
(psum n-butanol) Rf=0.2 (1'H-NMR(dff-DMF!O)Δ (Thos
) ppn+ 1, 1 8 (t s J = 7.5 H+ 6
H11,72~2.29 (m-4)T) 2
36 (m, 8FT) 3. o8(t,, J'=f
'i, 311. .

4It) 6.25-6.98 (m, 4T
() 7.38 (s, 2H) 111 (c++
-') 3500 1.605 1-2438117
5 1103 Examples 5. Preparation of compound (01) N-ethyl-1+-(3-sulfopropyl) ayu 1 nonnato 1 noum i 5.46 g was dissolved in water IQI/lci, 3
7 tiforma 1 no: /1.80m1,90 formic acid 0.20m
Add 1 and maintain the temperature at 35°C. a"+N-(3-sulfopropyl)aniline sodium salt 4.89 (y, f water IQm
(Dissolved in i, -t, -C added dropwise to carry out the reaction. Collect the bluish-white crystals d''.

Recrystallize with methanol to obtain white 4'-f N'-:r
, thyl-N-(3-sulfopropyl)7mino)2. Nyl-4-(N-(3-・sulfopropyl)amino)
Phenylmethane 2-J-IJ Al16.25gf
,Obtained. Yield: 590 cm TTIC (silica gel, 02N ammonia water 1'af
Iin-butanol)Rf=0.30'TI-)IMR(d@-1)Meen), l"pp
, (TM s)1.12 (t, J=7.61
(2,3H) 1. ! 59-2.1.6 (m+4
, TI) 2.36-2.77 (m, 6T
() 2.80-3.25 (+n, 4H)
3.49 (!E, 2H) 6.28~6
．． 56 (m.

4 Ti) 660-7.02 (m, 5
H) Engineering R (("l-') 3380 1600 1
2951201 1.025 Example 6 Preparation of Compound (Q,) 608 g of 3-methoxy-N-ethyl-N-(3-sulfopropyl)aniline sodium salt was dissolved in 10 ml of water, 1.80 ml of 37thiformin, 90% formic acid 0
．． Add 20 ml of N-ethyl-N-(3-sulfopropyl)aniline sodium salt while maintaining 35' OK.
．． 47 g was dissolved in LO ml of water and added, and 1,1-methylaniline was added to the water to carry out the reaction. The precipitated white crystals were collected and recrystallized from methanol to give 2-methoquino-4-fN-ethyl-N-(3-sulfopropyl)amicariphenyl-4'-εN'-edyl-N'-(3 -sulfopropyl)amine J phenylmethane disodium salt 7
．． Obtained 25gf.

Yield 61.5% rIJc (silica gel, 02N aqueous ammonia saturated n-butanol) Rf ~0.28' +1-NMR(da-DMSO)J
(T ), 4 S ) Ipm i, to (t, ,■ =7.7H,,6H
) 1.66 ~2. J, 5 (m.

4H) 2.39-2.78 (m, 8H) 2
．． F1a-3,33 (m, 4H) 3.55
(S, 2) () 4.11 (S, 3H) 6
．． 25-7.06 (m, 7ii) IR (preparation)
) 16013 1244 1206 110
0 Example 7゜1, Glucose No. 5 oxidase 200-11' position, Par A kindase 200
Unit, MBTH40,81ny, di(N-:
f-f-N-(3-sulfopropyl)amine]phenol-1-)methane disodium salt (Compound D)54.
Dissolve 2■ and use it as a coloring reagent.

Take 20 g of serum in a test tube, add the above coloring reagent i,
After adding Ome and reacting at 37°C for 10 minutes, pure water 2. Add Oml and measure the absorbance at 598 nm using a reagent blind control.
Glucose concentration in serum is calculated relative to Q+.

Standard Fertility Liquid Nigel Course 200■/de Sensitivity: 01111 i-1d

Claims (1)

[Claims] (In the formula, R1, nR is a hydrogen atom, a lower alkyl group, and a sulfoalkyl group having 2 to 3 carbon atoms is hydroxyl)
Indicates a substituent selected from the group consisting of Sunototsuζalkyl group increase→Tobito゛・Rokitasu★-TunruN, where R' and R' are carbon atoms of 2 to 3; The hydroxysulfoalkyl group is represented by R5R.
6 represents a substituent selected from the group other than a hydrogen atom, a Metquin group, a hydroxyl group, and a lower alkyl group) and its salts (in the formula, II and nN are hydrogen atoms, lower Represents a substituting group selected from the group consisting of an alkyl group, a sulfoalkyl group having 2 to 3 carbon atoms, or a hydroxysulfoalkyl group; A water-soluble diaminodiphenylmethane derivative represented by a hydroxysulfoalkyl group (R' and R' each representing a substituent selected from 11T selected from a rice atom, a methocino group, a hydroxyl group, and a lower alkyl group) and its salts are used as pigments. Composition for a peroxide pointer comprising as a forming substance