RU2654862C1 - Method for clearing disulfo derivative diamino triphenylmethane dye - Google Patents

Abstract

FIELD: technological processes.

SUBSTANCE: invention relates to a method for purifying impurity derivatives of a diaminotriphenylmethane dye from a disulfo derivative for subsequent use as a contrast agent in the detection of regional lymphogenous metastasis of malignant tumors in tumors of different localization, as well as in food, drugs, cosmetics, etc. Method for purifying a disulfo derivative of a diaminotriphenylmethane dye is proposed, which consists in adsorption of a disulfo derivative of a diaminotriphenylmethane dye on a sorbent element in the form of a plate of polydimethylsiloxane. Result is a disulfo derivative of diaminotriphenylmethane dye with a purity of at least 99.8 % with simplification of the purification technology.

EFFECT: proposed method is simple in execution, easily scalable and characterized by a low cost price.

8 cl, 3 dwg, 2 ex

Description

Technical field

The invention relates to the field of organic and medical chemistry, molecular biology, and in particular to a method for purification of impurities of a disulfonated diaminotriphenylmethane dye for subsequent use as a contrast agent for the detection of regional lymphogenous metastasis of malignant neoplasms in tumors of various localization, especially with breast cancer, stomach, colon, uterus, thyroid and prostate glands, melanoma. The high-purity disulfonated diaminotriphenylmethane dye can also be used in other fields, for example, in food products, pharmaceuticals, cosmetics, etc.

State of the art

Triarylmethane dye-based preparations are widely used in surgical practice for contrasting lymphatic vessels (for example, isosulfan blue, indigo carmine, methylene blue and others). Isosulfan blue is a triarylmethane dye and is used as a contrast agent for imaging lymphatic vessels, it is especially useful as a diagnostic agent for cancer. Limfazurin is a blue dye in pharmaceutical dosage form, available as a 1% (10 mg / ml) 5 ml solution in phosphate injection buffer. It is widely used in a procedure called "mapping of sentinel lymph nodes." It has been used to localize sentinel lymph nodes in patients with breast cancer. Isosulfan blue is cost-effective and safer to use than technetium 99 tons. Isosulfan blue is the structural isomer of sulfan blue; both compounds belong to the triarylmethane dye family. The main requirement for these drugs is good lymphotropicity and safety of use. Despite the wide variety of lymphotropic dyes, the drug based on isosulfan blue - “Lymphazurin injection 1%” is mainly used to contrast the “sentinel” lymph nodes.

Methods for producing (synthesizing) drugs using a blue dye are well known and diverse. Methods for the production of a drug (preparation) are distinguished by the use of various starting components and their proportions, the use of various synthesis options, the use of various methods of purification of the components and the final preparation, etc. The main task in this case is to obtain the final product (preparation) with a very high purity of the active substance - more than 99%.

The prior art method for the manufacture of isosulfan blue (active pharmaceutical ingredient) (patent US 7662992). The method allows to obtain isosulfane blue in almost pure form for use as a contrast medium. The aim of the invention is to provide a simple, safe and cost-effective method for producing isosulfan blue in a pure form, namely with a purity of 99.0% or more. In accordance with the invention, a simple procedure for the isolation of benzaldehyde-2,5-disulfonic acid, isoleiko acid and isosulfanic blue is provided. In this case, isosulfanic blue acids are purified by recrystallization from aqueous isopropyl alcohol / acetone to obtain isosulfonic blue acids with a purity of 99.5%. The final product, the isosulfan blue sodium salt, is obtained by the interaction of blue acids with a solution of sodium bicarbonate in an aqueous solution of acetone at a pH of more than 6.0. The reaction mass is filtered, obtaining the sodium salt of isosulfan blue, having a purity of more than 99.5%. In this method, when obtaining high purity of the obtained product, high performance liquid chromatography (HPLC) is used. This method is expensive and has low yields even when using preparative chromatographic systems.

In most cases, reprecipitation methods are used to purify compounds from impurities. Some dyes obtained in the form of aqueous solutions can be used in food products, drugs and cosmetics. When using dyes as additives in food products, pharmaceuticals and cosmetics, there are strict requirements for the purity of the dye used.

It is also known from the prior art to isolate a di-sulfonated diaminotriphenylmethane dye using various adsorbents containing polydimethylsiloxane (PDMS or PDMS). The principle of action of the adsorbents used is based on the extraction of the dye from the initial solutions containing the active agent.

In particular, the document CN 106582574 discloses a composition and method for producing an adsorbent including (by weight): 22-30 parts of pyrophyllite, 3-7 parts of polydimethylsiloxane, 2-6 parts of propylene glycol acetate esters, 1-5 parts of iron sulfate and 9-17 parts of 3-hydrochloric acid. The resulting adsorbent absorbs heavy metals, various types of dyes and other organic pollutants well and is used to purify aqueous media from these substances.

The prior art also knows a method of water purification using an adsorbent based on PDMS, which allows the extraction of organic solvents and oils from the aqueous phase (Choi et al., 2011). To do this, the surface of the pressed sugar is coated with polydimethylsiloxane, then the resulting structure is placed in water until the sugar is completely dissolved and thus a sponge structure of polydimethylsiloxane is formed. Such a structure has a large surface area and is capable of adsorbing organic solvents and oils in an amount up to four times the mass of polydimethylsiloxane itself.

Thus, well-known sources of information disclose compositions / designs of adsorbents in which PDMS is present as one of the components, while the action of these adsorbents is aimed at the isolation (extraction) of organic or inorganic substances from the aqueous phase. The prior art did not identify sources of information from which it would be known to use PDMS for the purification of disulfonated diaminotriphenylmethane dye from their impurities.

The claimed solution is characterized by a different principle of action of the adsorbent used, which is a plate of PDMS, based on the extraction from the initial solutions of the active component - dye without impurities, which remains in the initial aqueous solution.

Disclosure of invention

An object of the present invention is to provide a new method for purifying a disulfonated diaminotriphenylmethane dye.

The technical result of the claimed invention is to obtain a disulfonated diaminotriphenylmethane dye with a purity of not less than 99.8% while simplifying the purification technology. The inventive method is simple to implement, easily scalable and is characterized by low cost.

The problem is solved by the claimed method, which includes the following steps:

- a sample of a disulfonated diaminotriphenylmethane dye is dissolved in an aqueous solution of 0.25-1.0 M NaCl or KCl;

- the sorbing element PDMS is immersed in the resulting solution, which is a plate with a certain geometry and size, ensuring the adsorption of the dye for 12-24 hours at room temperature with stirring; wherein the plate is taken with a thickness of at least 0.2 cm and an area of at least 10 cm ² and for cleaning a solution containing 5 g of dye.

- then the solution is removed, the sorbent is washed with equal portions of a solution of 0.25-1.0 M NaCl or KCl, completely covering PDMS, to remove residual impurities until the wash water is clear;

- the main substance is washed from the PDMS plate with a solution of 0.25-1.0 M NaCl or KCl with the addition of 0.01-0.05 M NaOH or KOH with a pH of 6.5-7.5, the pH of the resulting solution is adjusted to these values with aqueous a solution of 0.01-0.05 M HCl, with stirring, followed by evaporation of the solution and drying the resulting precipitate. In this case, a disulfonated diaminotriphenylmethane dye is obtained with a purity of not less than 99.8%.

Brief Description of the Drawings

The invention is illustrated by drawings, where in FIG. 1 shows an exemplary embodiment of a PDMS plate. This plate allows cleaning up to 75 g of the compound; in FIG. 2 shows a chromatogram of a sample of the compound before purification, containing impurities in an amount of 4.45 wt. %; in FIG. 3 presents a chromatogram of a sample of the compound after purification by the claimed method, the impurity content of which is 0.2 wt. %, the content of the main substance of 99.8 wt. % (example 1).

The implementation of the invention

Below is a more detailed description of the proposed method, which does not limit the scope of claims of the claimed invention, but demonstrates the possibility of carrying out the invention with the achievement of the claimed technical result.

The method is intended for the purification of disulfonated diaminotriphenylmethane dye, namely potassium bis (4 - {[4- (diethylamino) phenyl] [4- (diethyliminium) cyclohexa-2,5-diene-1-ylidene] methyl} -6-hydroxybenzene-1 , 3-disulfonate) (potassium salt of oxysulfane blue) or sodium 4 - {[4- (diethylamino) phenyl] [4- (diethyliminimyl) cyclohexa-2,5-diene-1-ylidene] methyl} -6-hydroxybenzene-1 , 3-disulfonate (oxysulfane blue sodium salt). The main impurity of the salts of the disulfonated diaminotriphenylmethane dye is a compound of the same structure without an ethyl group — a compound of formula (2). According to the results of purification of the initial solution, a compound of the formula (1) is obtained.

To remove impurities, samples of the compounds are dissolved in an aqueous solution of 0.25-1.0 M NaCl for the sodium salt of the dye and 0.25-1.0 M KCl for the potassium salt of the dye. The amount of substance (sample) for dissolution is governed by its solubility (up to 75 g per 1 liter of solution), it is preferable to prepare a solution at the rate of 1 g of sample per 14-27 ml of aqueous saline. Then, a plate of PDMS is immersed in the resulting dye solution. In this case, the plate is chosen with geometric dimensions that ensure maximum adsorption of pure dye. This property is achieved when using a plate with a thickness of at least 0.2 cm and an area that is calculated based on the quantitative content of the dye in the solution, where a plate with an area of at least 2 cm ^{2 is} used to clean 1 g of the dye. It is preferable to use polydimethylsiloxane (PDMS) Sylgard® 184 Silicone Elastomer manufactured by Dow Corning on the basis that about 5 g of PDMS are taken per 5 g of the compound to be purified and a plate with an aspect ratio of 50 × 25 × 1 (L × W × H) is formed at this working surface of the plate can be smooth, corrugated or rough, to increase the adsorption surface. The resulting system at room temperature is incubated for 12-24 hours with constant stirring. Mixing can be carried out on a magnetic stirrer at a stirring speed of 50-100 rpm. After the specified time, the solution is removed, and the sorbent plate is washed in portions of 0.25-1.0 M NaCl or KCl to remove impurities. For this, the PDMS plate is poured with a solution of NaCl or KCl in portions that completely cover the PDMS. Rinsing is continued until the saline solution is clear. Then the basic substance adsorbed on the PDMS plate is washed out, for which a solution of 0.25-1.0 M NaCl or KCl with the addition of 0.01-0.05 M NaOH or KOH is used, the pH of the solution is adjusted to pH 6.5-7, 5 through the use of an aqueous solution of 0.01-0.05 M HCl, pouring a plate on it until it is coated, and the resulting solution is kept for 2-4 hours at room temperature with constant stirring. As a result of changing the pH, the distribution coefficient of the basic substance (logD) changes and the adsorbed compound characterized by the formula (1) is released into the solution. In this case, the solution stains blue and the PDMS plate enlightens. Then the plate is removed, the solution is evaporated at a temperature not exceeding 40 ° C, preferably on a rotary evaporator, the resulting precipitate is dried at a temperature not exceeding 40 ° C.

As PDMS, a material with the following characteristics is preferably used:

In this case, the key parameter on which the sorption activity of PDMS depends, is the density of the polydimethylsiloxane used, which should be no more than 965 kg / m ³ , while to ensure the preservation of the shape of the plate, the viscosity should be at least 3.5 pascal-seconds. The optimal values of these parameters are achieved in the manufacture of PDMS plates in accordance with Dow Corning's Sylgard® 184 Silicone Elastomer. To control the process of adsorption and desorption of the dye, the key parameter is the color of PDMS.

It was proposed to use a glass container as a device for purifying a disulfonated diaminotriphenylmethane dye to prevent sorption of the main substance on its walls with a magnetic stirrer and a plate made of PDMS made with the possibility of immersing its “working” part (2 working surfaces) in the volume of the container with simultaneous fixation on the edge of the tank (see Fig. 1). In this design, the volume of the container for accommodating the initial solution and the geometric dimensions of the working part of the plate from PDMS, which is intended for contact with the solution to be cleaned, are selected. The ability to fix the plate can be ensured by the presence of an additional part made in the form of a continuation of the plate made of PDMS having a geometry that provides the convenience of its capture and fixation on the edge of the tank, for example, by means of elements protruding outside the tank, provided with grooves in the area of their interface with the tank. Moreover, the profile of the side surface of the working part of the plate has a wave-like character to increase the surface area of PDMS.

All reagents used are commercially available, all procedures, unless otherwise specified, were carried out at room temperature or ambient temperature, that is, in the range from 18 to 25 ° C.

Example 1

The claimed invention was implemented using a device (Fig. 1), in which a beaker with a volume of 1 liter, a height of 15 cm and a diameter of 10 cm and a plate of PDMS with a working part of 10 cm × 5 cm × 0.2 cm in weight of 10 were used g.

50 g of oxysulfane blue sodium salt was dissolved in 1 L of an aqueous solution of 0.25 M NaCl, incubation was carried out with a sorbing element PDMS for 12 hours at room temperature. Then the solution was removed, the sorbent was washed with equal portions of a 0.25 M NaCl solution until the wash water was clear. The basic substance was washed with 1 l of a solution of 0.25 M NaCl with the addition of 0.05 M NaOH. The pH of the resulting solution was adjusted to pH 6.5-7.5 with an aqueous solution of 0.05 M HCl. The resulting solution was evaporated on a rotary evaporator at a temperature of 40 ° C and the resulting precipitate was dried at 40 ° C in an air thermostat. The impurity content of the obtained oxysulfane blue was determined by high performance liquid chromatography using a stainless steel column with a size of 250 mm × 4.6 mm filled with a sorbent C18 with a particle size of 5 μm (type Luna C 18 (2), cat. No: 00G-4252- E0, Phenomenex or similar); as the mobile phase, methanol was used with solution A containing 6.8 g of potassium dihydrogen phosphate and 17.4 g of disodium hydrogen phosphate dihydrate per 1 water, pH 7.0, (50/50 v / v) with a flow rate of the mobile phase of 1, 0 ml / min, thermostat temperature of the column is 35 ° C. Compounds were detected at a detector wavelength of 638 nm (SF), the volume of the injected sample was 10 μl, and the chromatographic time was 35 min; the retention time of the impurity peak is about 17.5 minutes, the oxysulfane blue sodium salt is about 22 minutes.

The impurity content before purification was 4.45 wt. % (Fig. 2), after purification - 0.2 wt. % (Fig. 3). The purity of oxysulfane blue was 99.8 wt. % The result was 44 g of oxysulfane blue sodium salt. The yield was 88 wt. %

Example 2

Carried out in a similar manner (example 1), but instead of an aqueous solution of 0.25 M NaCl, 0.7 M was used and the incubation time was 20 hours, and a solution of 0.25 M NaCl with the addition of 0.02 M NaOH was used to wash the dye.

The content of impurities before purification was 5.5 wt. %, after purification - 0.1 wt. % The purity of the oxysulfane blue sodium salt was 99.9 wt. % The result was 44.5 g of oxysulfane blue sodium salt. The yield was 89 wt. %

Example 3

The purification of the potassium salt of oxysulfane blue is carried out in a manner analogous to example 1, only an aqueous solution of KCl was used as a solvent and a washing solution. The basic substance was washed with a KCl solution with the addition of KOH.

The impurity content before purification was 7.5%, after purification - 0.1%. The purity of the potassium salt of oxysulfane blue was 99.9%. The result was 44.5 g of potassium salt of oxisulfan blue. The yield was 89%.

Thus, the proposed method for purifying a disulfonate diaminotriphenylmethane dye, which consists in adsorbing a disulfonate diaminotriphenylmethane dye on a sorbing element in the form of a plate of polydimethylsiloxane, is simple, easily scalable, and a disulfonate diaminotriphenylmethane dye with a purity of at least 99.8% is obtained.

Claims (8)

1. A method of purifying a disulfonated diaminotriphenylmethane dye, comprising mixing the dye with an aqueous solution of 0.25-1.0 M NaCl or KCl, immersing in the resulting solution a sorbing element in the form of a plate of polydimethylsiloxane, keeping the plate for a period of time providing adsorption of a pure disulfonated diaminotriphenylmethane dye from the solution into the plate material, followed by removal of the solution, after which the plate is washed from the impurities contained in the solution, and the pure dye is removed from the pl astins, while the plate is taken with a thickness of at least 0.2 cm and an area that is calculated based on the quantitative content of the dye in the solution, where a plate with an area of at least 2 cm ^{2 is} used to clean 1 g of the dye. 2. The method according to claim 1, characterized in that the potassium bis (4 - {[4- (diethylamino) phenyl] [4- (diethyliminium) cyclohexa-2,5-diene-1-ylidene] is used as a disulfonated diaminotriphenylmethane dye. methyl} -6-hydroxybenzene-1,3-disulfonate) or sodium 4 - {[4- (diethylamino) phenyl] [4- (diethyliminimyl) cyclohexa-2,5-diene-1-ylidene] methyl} -6-hydroxybenzene -1,3-disulfonate. 3. The method according to claim 1, characterized in that an aqueous solution of 0.25-1.0 M NaCl or KCl for mixing with the starting compound is taken at the rate of 14-27 ml per 1 g of dye. 4. The method according to claim 1, characterized in that the aging of the plate in the solution is carried out for 12-24 hours at room temperature with stirring of the solution. 5. The method according to claim 1, characterized in that the washing of the plate from the impurities contained in the solution is carried out by repeatedly immersing the plate in a solution of 0.25-1.0 M NaCl or KCl until the wash water is transparent. 6. The method according to claim 1, characterized in that to extract the pure dye from the plate material, it is immersed in a solution containing 0.25-1.0 M NaCl or KCl and 0.01-0.05 M NaOH or KOH, after whereby the pH of the solution is adjusted to a pH of 6.5-7.5 by adding an aqueous solution of 0.01-0.05 M Hcl, followed by evaporation of the solution and drying the resulting precipitate. 7. The method according to claim 1, characterized in that polydimethylsiloxane with a density of not more than 965 kg / m ³ and a viscosity of at least 3.5 Pa · s is used as polydimethylsiloxane. 8. The method according to claim 1, characterized in that to increase the adsorption area of the plate has a corrugated or rough surface.

Images