Fluorination method for synthesis of 2-[18f]-fluoro-2-desoxy-d-glucose

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing a protected fluorinated glucose derivative, involving reaction of a tetraacetylmannose derivative with a fluoride, distinguished by that the reaction is carried out in a solvent which contains water in amount of more than 1000 parts per million and less than 50000 parts per million. Preferably, the protected fluorinated glucose derivative is 2-fluoro-1,3,4,6-tetra-O-acetyl-D-glucose (tetraacetylfluroglucose or pFDG), the tetraacetylmannose derivative is 1,3,4,6-tetra-0-acetyl-2-0-trifluoromethanesulphonyl-β-D- mannopyranose (tetraacetylmannose triflate), the solvent is acetonitrile, the fluoride is a fluoride ion with a potassium counter-ion, and a phase-transfer catalyst, such as 4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo-[8,8,8]-hexacosa, is added to the fluoride.

EFFECT: improved method.

14 cl, 2 tbl, 3 dwg, 3 ex

 

The invention relates to a method of fluorination derivatives of sugars and, in particular, the invention relates to the production of fluorinated glucose. The method is especially suitable for the preparation of derivatives of sugars, fluorinated radioactive fluorine used in such techniques as positron emission tomography (PET).

Cooking methods of tracers labeled with [18F], which are used in PET, one of the most important factors is the total unadjusted output of the synthesis. It depends not only on the chemical out of the way, but from the time of synthesis, which is critical due to the relatively short half-life of [18F], constituting 109,7 minutes.

[18F]-fluoride ion is usually prepared in the form of an aqueous solution obtained by cyclotron irradiation of the target molecules of water containing [18About]. For the conversion of [18F]-fluoride in reactive nucleophilic reagent suitable for use in the nucleophilic introduction of radioactive isotopes, used a variety of operations. As in the fluorination reactions of non-radioactive fluorine, these operations included the elimination of water from [18F]-fluoride ion and the creation of a suitable counterion (Handbook of Radiopharmaceuticals 2003 Welch & Redvanly, eds. Ch.6 pp.195-227). Sateriale nucleophilic radiotolerance carried out in anhydrous solvents (Aigbirhio et al., 1995, J. Fluor. Chem. 70 pp.279-87). The separation of water from fluoride ion is called cooking "naked" fluoride ion. The presence of significant quantities of water, as I believe, leads to solvation of fluoride ions, shielding fluoride ion, which prevents the carrying out nucleophilic attack on a protected precursor of sugar. Thus, in the art Department of water is surgery necessary to improve the reactivity of the fluoride ion and avoid getting side-hydroxylated products formed in the presence of water (Moughamir et al., 1998 Tetr. Letts. 39 pp.7305-6).

In U.S. patent 6172207 relating to the method of synthesis of compounds labeled with [18F], such as [18F]-Tordesillas ([18F]-FDG), emphasized the need for absolutely anhydrous fluorinating agent, which is obtained by adding acetonitrile in aqueous solutions, followed by azeotropic evaporation to dryness.

The most widely used method for the synthesis of [18F]-FDG is the method proposed Hamacheret al., J. Nucl. Med. 27:235-238 (1986), which comprises carrying out the reaction of 1,3,4,6-Tetra-O-acetyl-2-O-trifloromethyl-β-D-mannopyranose with [18F]-fluoride in an anhydrous solvent.

The methods currently used for the preparation of derivatives of sugars, labeled with [18F], have a number of nedostatki is; one of them is the fact that complete separation of residual water from fluoride ion and solvent takes some time and, therefore, affect the overall unadjusted output of the synthesis. In addition, the need to remove all residual water leads to increased synthetic and mechanical complexity of any automatic synthesizing device. For example, synthesis may require a higher number of cycles of drying, while to perform the synthesis in synthesizing device may need to install a more powerful heater.

Moreover, usually it is quite difficult to ensure good reproducibility of the reaction radiotolerance. This is due to the frequent presence of small amounts of residual water in the solvent (e.g., in the amount of approximately 1000 parts per million), and total unadjusted output of synthesis varies greatly depending on the amount of residual water present during the reaction injection of radioactive indicator. It was found that it is possible to maintain the water content amounting to 1500 ppm +/-200 ppm, i.e. with a deviation of 15%. At 750 parts per million is the absolute deviation of the water content will lead to a doubling of the percentage of deviation.

The authors found that ftoridov is of derivatives of sugars is not necessarily carried out in anhydrous conditions. Indeed, if carefully regulate the water content in the reaction mixture, it actually increases the radiochemical purity (and hence total output) obtain derivatives. This is particularly unexpected, given the special attention that at the current level of technology has been conducting the reaction in anhydrous conditions.

Thus, the first aspect of the present invention relates to a method for producing a fluorinated derivative of sugar, including the interaction of non-fluorinated derivative of sugar with fluoride, characterized in that the above reaction is carried out in a solvent containing water in excess of 1000 parts per million and less than 50,000 parts per million.

The method proposed according to the present invention has significant advantages over existing methods of the prior art. First, it was found that the yield of the reaction is not only not diminished, but, on the contrary, increases in the presence of controlled quantities of water.

Secondly, because the water content in the reaction mixture exceeds 1000 parts per million, to ensure the presence of an appropriate quantity of water in the reaction mixture becomes much easier (for example, intentionally introducing contaminating quantities of water in the solvent for the reaction, which means the greater the Yu reproducible reaction conditions.

Thirdly, it eliminates several stages of drying used in the existing level of technology that can reduce the overall cost of the method in relation to the cost of reagents, and the cost of synthesizing device. Also believe that the simplification of the way will have a positive impact on the overall reliability of the method.

In the present description, the term "non-fluorinated derivative of sugar" refers to sugar type of polysaccharide, oligosaccharide, disaccharide or monosaccharide in which one of the groups HE substituted useplease group, which may be associated with a solid substrate, for example as described in WO-A-03/002157. The method proposed according to the present invention, especially suitable for the fluorination of monosaccharides, such as glucose, fructose, ribose, arabinose, mannose or galactose.

In "protected non-fluorinated derivative of sugar," the rest of the group HE sugar protected by suitable protective groups.

The term "fluorinated derivative of sugar" refers to sugar type of polysaccharide, oligosaccharide, disaccharide or monosaccharide, such as glucose, fructose, ribose, arabinose, mannose or galactose, in which one of the groups HE is replaced by fluorine atom.

In "protected fluorinated derivative of sugar," the rest of the group HE sugar protected by suitable protective groups.

Suitable protective groups used to protect derivatives of sugars in accordance with the present invention, are known in the art and described, for example, in the publication "Protecting Groups in Organic Synthesis", Theodora W.Green and Peter G.M.Wuts published by John Wiley & Sons Inc. The choice of specific protective group depends on the intended use of fluorinated product, but, for example, hydroxyl group can be protected by converting it to an alkyl or aromatic ester group, for example, by reaction with alcantarillados, such as acetylchloride. In the alternative case, the hydroxyl group can be converted into ester group, such as alkyl or benzyl esters.

Preferably, both the raw materials and products of the reaction are protected derivatives of sugars.

Suitable useplease groups are well known in the art and include toluensulfonate and methansulfonate group. However, particularly preferred useplease group is triftormetilfosfinov (triflate) group.

The fluorination reaction is usually a reaction of nucleophilic substitution, and the substitution useplease group fluorine may cause inversion of the stereochemical structure of sugar flowing through the SN2 mechanism. Thus, the original neveryou the Noe-derived sugar is often a derivative of sugar, which is different from the product.

The preferred product is a protected fluorinated derivative of glucose, which can be obtained from the corresponding derived mannose, for example derived from tetracylines.

The reaction is particularly suitable for the preparation of 2-fluoro-1,3,4,6-Tetra-O-acetyl-D-glucose (tetraarylporphyrin or pFDG) of 1,3,4,6-Tetra-O-acetyl-2-O-trifloromethyl-β-D-mannopyranose (triflate tetracylines).

Suitable solvents include aprotic organic solvents such as acetonitrile, dimethylformamide, dimethylsulfoxide, tetrahydrofuran, dioxane, 1,2-dimethoxyethane, sulfolane or N-methylpyrrolidinone or any mixture of these solvents. However, it was found that the most preferred solvent for the reaction is acetonitrile.

Despite the fact that the increase of the yield of the reaction is reached due to the inclusion in the solvent is at least 1000 parts per million, but less than 50,000 parts per million of water, further increasing output was achieved by the addition of from 1000 to 15,000 parts per million of water. The best results were obtained when using the solvent, the water content of which was approximately from 2,000 to 7,000 parts per million, preferably from 2500 to 5000 parts per million. In one of the examples Rea is Itachi the preferred water content is from 3000 to 6000 parts per million.

In the present description to mean water content in a particular solvent, the term "parts per million" means micrograms of water/gram.

Proper concentration of water in the solvent can be achieved either by drying the wet solvent to achieve the desired water content, or by adding a suitable amount of water in the dry solvent. Fluoride can be obtained in aqueous solution and in this case the solution of the fluoride with the desired water content can be obtained by repeated addition of a solvent followed by evaporation of the mixture solvent/water or a dilute aqueous solution of fluoride in the desired organic solvent. The water content of the solvent can also be reduced by using an adsorption resin, such as functionalized polystyrene resin, for example epoxy resin, methylisocyanate resin or functionalized resin-based anhydrides of acids designed to remove water from a solution of fluoride. Suitable resins are commercially available products, for example, supplied by Novabiochem. Specifications absorbing resin can be improved by using a suitable catalyst, for example, 4-dimethylaminopyridine (4-DMAP).

In this example, the implementation stage drying can be carried out by mixing the absorbent resin with a solution of fluoride in the container and the subsequent separation of the absorbent resin by filtration. In alternative and is especially suitable if absorbing resin used in the automatic synthesizing device, absorbing resin may be in the vessel, through which pass the fluoride solution. A solution of fluoride can pass through an adsorption resin continuous flow, for example with a speed of from 0.1 to 100 ml/min, or in periodic mode, maintaining the solution in contact with absorptive resin in the course of time sufficient to dry.

This use of absorptive resin is new; therefore, in accordance with the following aspect of the present invention, a method for reducing the water content in the solution radiofonica, in particular in solution [18F]-fluoride, which includes the specified contact of the solution with an adsorption resin. It is convenient to obtain the solution of the fluoride in an aprotic organic solvents, such as acetonitrile, dimethylformamide, dimethylsulfoxide, tetrahydrofuran, dioxane, 1,2-dimethoxyethane, sulfolane and N-methylpyrrolidinone, the most suitable solvent is acetonitrile.

The reaction can be conducted in solution or, alternatively, non-fluorinated derivative of sugar may be associated with the solid substrate with the formation of the vector resin-linker (RVL) of the formula (I):

SOLID SUBSTRATE-LINKER-X-Protected not tonirovanoe derived sugar (I)

where the solid substrate is any suitable solid substrate;

protected non-fluorinated derivative of sugar defined above;

X is a group promoting nucleophilic substitution at a particular facility is protected non-fluorinated derivative of sugar, for example-SO2O-;

the linker is any organic group, which is sufficient for separation of the reactive center and structure of solid surfaces with the aim of creating maximum reactivity, for example containing from zero to four aryl groups (e.g. phenyl), and/or C1-C6is an alkyl or halogenation (especially alkyl fluoride) chain, and possibly from one to four additional functional groups such as amide or sulfonamidnuyu group.

System RVL is described in detail in WO-A-03/002157, which also describes suitable linkers.

RVL formula (I) enter into contact with a solution of fluoride, which leads to the elimination of sugar from the solid substrate with the formation of protected non-fluorinated derivative of sugar.

Suitable solid substrate is also described in WO-A-03/002157 and include polymers such as polystyrene (which may be blogbrevity, for example, polyethylene glycol), polyacrylamide, or polypropylene, or glass, or silicon with Nan who built on it a coating of the specified polymer. Alternatively, for example, there may be used resin such as those described in WO-A-03/002157. The solid substrate may be in the form of small isolated particles, for example balls or sticks, or in the form of a coating on the inner surface of the cartridge or MicroSCADA. Carrying out the method according to the present invention, the solid substrate allows to obtain the product in a pure form, without conducting an additional phase separation. This is particularly advantageous if fluoridation is radiotolerans, because any time saved in the manufacture of the product increases uncorrected radiochemical yield.

The reaction is usually carried out at a temperature of from 5 to 180°C, but in particular from 75 to 125°C.

The method proposed according to the present invention can be implemented as part of an automated synthesis. This is feasible if the reaction proceeds in solution, or if non-fluorinated sugar is associated with the solid phase.

The fluoride that is put into interaction with non-fluorinated derivative of sugar, can be an ionic compound and can be paired with any suitable counterion. However, it is important to note that the counterion must have sufficient solubility in the reaction solvent, in order to maintain the solubility of the photo is IDA. Thus, suitable counterions include large, but soft metal ions such as rubidium or cesium, or, alternatively, nonmetal ions such as tetraalkylammonium ions and tetraalkylammonium. Potassium ions can also be used as counterions, but in this case, to increase the reactivity of the fluoride type catalyst phase transfer, such as 4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo-[8,8,8]-hexacosane (sold under the trademark Kryptofix™ 2.2.2)in order to improve the solubility of potassium salts in organic solvents.

The method proposed according to the present invention is well suited for obtaining radiotolerance derivatives, in particular derivatives labeled with [18F], and therefore, the fluoride may include [18F]-fluoride ion.

As was briefly noted above, [18F]-fluoride ion can be obtained by irradiation of the target molecules of water containing [18About], and this operation may represent the initial stage of the method proposed according to the present invention.

The method according to the present invention is particularly suitable for receiving radiotolerance derivatives of sugars, for example, [18F]-pFDG, which can then be unprotected with the formation of compounds such as [18F]-FDG, a well-known tracer for PE is. Removing protection can be an additional step of the way. If the protective group of the obtained fluorinated sugar is an ester, for example a derivative of acetyl, removing protection can be acid or alkaline hydrolysis.

Other additional stages include the removal of excess [18F]-fluoride from the solution and removing the organic solvent. An excess of [18F]-fluoride can be removed by any standard method, such as ion exchange chromatography or solid-phase absorbers. Suitable ion exchange resins include BIO-RAD AG 1-X8™ and Waters QMA™and suitable solid phase materials include aluminum oxide.

The organic solvent can be removed by evaporation at elevated temperature in vacuum or by passing a stream of inert gas, such as nitrogen or argon through the solution.

The final product of these stages, i.e. isotopic indicator labeled with [18F], can be manufactured in the form of a composition for administration to a patient, for example in the form of an aqueous solution, which can be prepared by dissolving labeled with [18F] tracer in sterile isotonic saline solution, which may also contain up to 10% of a suitable organic solvent, for example ethanol, or, alternatively, in a suitable buffer solution such as phosphate the output buffer. In the song, you can add other additives, such as ascorbic acid, which reduces the radiolysis.

As has already been noted, particularly preferably a compound that can be obtained by the method according to the present invention, is a [18F]-pFDG, and therefore, the second aspect of the present invention relates to a method for preparation of [18F]-pFDG, and this method involves the reaction of the triflate tetracylines with [18F]-fluoride and differs in that the fluoride is dissolved in a solvent containing water in excess of 1000 parts per million and less than 50,000 parts per million. In one example implementation of this aspect of the present invention, triplet tetracylines (1 equivalent) interacts with [18F]-fluoride in the presence of Kryptofix™ 2.2.2 (from 0.9 to 1.1 molar equivalents; preferably from 0.98 to 0.99 molar equivalents) and potassium carbonate (0.4 to 0.6 molar equivalents; preferably from 0.50 to 0.60 molar equivalents) in acetonitrile containing water in excess of 1000 parts per million and less than 50,000 parts per million.

Preferred features of the present invention discussed in detail above for the first aspect of the invention. In particular, the method may include the initial step for [18F]-fluoride obleceni the m target molecules of water, contains [18About], and subsequent phase transformation [18F]-pFDG [18F]-FDG acid or alkaline hydrolysis.

Hereinafter the invention will be described in more detail with reference to examples and drawings where:

Figure 1 shows a graph of the dependence of the radiochemical purity of [18F]-pFDG the water content in the solvent.

Figure 2 is a plot of the formation of [18F]-pFDG and protected glucose in the process of introducing a tracer using vector resin-linker.

Figure 3 is a graph of the dependence of the radiochemical purity of [18F]-pFDG the water content in the solvent during automated synthesis.

Example 1. The impact of changes in water content on the introduction of the tracer [18F] sugar molecules

In this example, used three different ways of introducing tracer18F-and explored the impact of changes in water content in the reaction mixture.

a) the Introduction of the tracer using a vector resin-linker (RVL)

Ion18F-introduced in Tracerlab MX™, and the system was dried using conventional drying method used in obtaining 2-[18F]-fluoro-2-deoxyglucose. To increase the solubility of fluoride in acetonitrile was used Kryptofix™ 2.2.2/potassium carbonate. After drying and subsequent dissolution of fluoride and in which lonitrile selected sample solution drained 18F-in acetonitrile for analysis of water content, which was measured with micrometre Karl Fischer. If necessary to achieve a water content in excess of the concentration obtained when dried, was added an additional amount of water.

In the cartridge Hi Trap® with a capacity of 1 ml Packed approximately 370 mg predecessor protected mannose associated with a solid-phase substrate, with the substitution of 0.003 mmol/g One end of the cartridge is connected by hinges with piston syringe. The other end of the cartridge was connected to a bottle, filled with N2and provided with an outlet opening, a closed molecular sieve. For heating cartridge used a spray gun hot air; external temperature cartridge was 80°C.

Then through the system missed a 6×0.5 ml of dry acetonitrile to remove any impurities and water (naturally present in the resin, for example, due to incomplete drying) and then acetonitrile was dropped. Bottle for autotbar samples containing the dried solution18F-(450 μl)was inserted into the machine at the right places. Then the piston of the syringe moved the dried solution fluoride reciprocating manner at a flow rate of 180 ál/min for 5 cycles. Dried fluoride interacted with the solid-phase precursor mannose-vysvobojdenie protected derivative of [ 18F]-2-deoxyglucose (which after removal of the protection form 2-[18F]-2-deoxyglucose).

Then out of the vial for autotbar samples were taken the sample volume of 5 μl, which was investigated using thin-layer chromatography (TLC); the sample was placed on a plate of silica gel 60 F254 and showed a mixture of acetonitrile with water taken in relation to 90/10. Radiochemical purity was determined by means of the device of the instant imaging Perkin Elmer.

b) Introduction of the tracer in triplet tetracylines

Preparing a solution of 32 mg K2CO3600 μl of H2O chromatographic purity with the addition of 150 mg Kryptofix™ 2.2.2 dissolved in 2.5 ml of acetonitrile. In a reactor made of glass carbon, were placed 0.6 ml of this solution and approximately 40 MBq (megabecquerel)18F-in water, enriched18Acting Automatic control of the heater was set at around 95°C and the reaction vessel was heated for 35 minutes to dry fluoride. The water and the acetonitrile evaporated in a stream of nitrogen.

To accelerate the azeotropic removal of water from fluoride was introduced, dividing into three portions, 1 ml of acetonitrile in 2-minute intervals; the first addition was made 20 minutes after the start of the drying operation. After 35 minutes, the heater was switched off and the reaction vessel was cooled by compressed is ozdoba outside the reaction vessel to a temperature of approximately 45°C.

Then to the dried fluoride was added 25 mg of mannose triflate in 2.0 ml of CH3CN and stirred. Automatic control of the heater was set at around 85°C. After 2 minutes after reaching the set temperature selected sample for TLC. The heater was switched off and the reaction vessel was cooled by compressed air stream to a temperature of approximately 45°C.

The sample for TLC was placed on a strip with silica gel and showed a mixture of acetonitrile with water, taken in the ratio 95:5. Radiochemical purity was determined by means of the device of the instant imaging Perkin Elmer. Then took the lid of the reaction vessel and selected a sample volume of 50 ál for analysis of water content, which was carried micrometre Karl Fischer.

(C) Automated introduction of tracer

The introduction of the tracer was performed using a prototype platform for automated synthesis, including 25 three-way valves, built-in heated reaction vessel and disposable polypropylene tape. Flow channel of the cartridge also allowed to do the cleaning of the intermediate product or final product using solid-phase extraction.

Originally fluoride was recorded on the cartridge cabballero mass analyzer (QMA), and it was suirable solution provided is 20 mg Kryptofix™ 2.2.2, 4,1 mg K2CO3, 320 μl of CH3CN, 80 μl of H2O. the mixture is Then dried at 105°C/120°C for approximately 6 minutes in a stream of nitrogen and re-dissolved in 1.5 ml of a solution of the triflate tetracylines in acetonitrile concentration of approximately 20 mg/ml

Reaction injection tracer was carried out at a reaction temperature equal to or 105°C or 120°C for either 90 or 270 seconds. After the introduction of the tracer 2-[18F]-2-desoxyglucose analyzed by way of TLC. Plate for TLC was a plate of silica gel 60 F254; the chromatogram showed a mixture of 95% acetonitrile and 5% water. Radiochemical purity was determined by means of the device of the instant imaging Perkin Elmer.

The results of the three experiments of Example 1 are shown in figure 1, from which it is clear that the radiochemical purity of the product is relatively low, if the water content in the solvent is less than 1,000 parts per million, but it is much better if the water content in the solvent is from 1000 to 5000 parts per million. From the graph we see that the optimal concentration of water in the solvent is in the range from 2,000 to 7,000 parts per million.

Example 2. The correlation between the formation of [18F]-pFDG and protected glucose in the process of introducing a tracer

The introduction of [18F] RVL conducted in acetonic the sludge in the presence of Kryptofix™ 2.2.2, potassium carbonate and various quantities of water. After the introduction of the indicator resulting mixture was subjected to liquid chromatography high pressure (ghvd) with reversed-phase, gradient developer: from a mixture of 90% solvent A:10% solvent B (solvent A=0.1% solution triperoxonane acid in water; solvent B=0.1% solution triperoxonane acid in acetonitrile) to 5% And 95%, for 10 minutes at a speed of 1 ml/min, using a column Phenomenex Luna 5 μm C18(4.6 mm × 150 mm). Were identified and correlated integral peaks corresponding protected glucose when the delay time is 3 minutes, and protected FDG with time delay, equal to 6.6 minutes (mainly due to the presence of [19F]-FDG, the concentration of which is proportional to the concentration of [18F]-FDG).

In General, I believe that the presence of large quantities of water in the reaction mixture leads to the formation of large quantities of protected glucose (instead of [18F]-pFDG), resulting nucleophilic substitution triflate group. Thus, suppose that the graph of the concentration of [18F]-pFDG concentration protected derivative of glucose in the mixture of reaction products must have a negative slope, and the large concentration of water should lead to the formation of large concentrations of protected glucose, and the smaller is oncentrate [ 18F]-pFDG.

However, studies of the introduction of the tracer in triplet tetracylines associated with resin, showed that there is a strong positive correlation (see Figure 2) between the two peaks on the chromatogram ghvd. This indicates that the presence of high concentrations of water slows down the formation of both products.

Example 3. Automated synthesis of 1,3,4,6-Tetra-O-acetyl-2-fluoro-β-D-mannopyranose

The sampling of radioactive reaction mixture at the beginning of the injection of the radiotracer was problematic. Thus, water content, and radiochemical purity was measured by using instant thin-layer chromatography (MTSH) upon termination of the reaction the introduction of the radiotracer. Then the water content at the beginning of the reaction the introduction of the radiotracer was calculated by determining the ratio of binding water, as described below.

The experiment on introduction of the radiotracer

Synthesis of 1,3,4,6-Tetra-O-acetyl-2-fluoro-β-D-mannopyranose was measured using an automatic synthesizing device which may be inserted a disposable cassette. This tape includes 25-valve disposable cassette including a variety of bottles containing reagents, as well as syringes and space for installation of solid-phase extraction cartridges.

Then did p is the sequence of the synthesis, including fixing of approximately 50 MBq of [18F]-fluoride in 2 ml of water in the cartridge Waters Access PlusQMA (carbonate form) and subsequent elution of the cartridge with a solution of Kryptofix and carbonate in acetonitrile/water (Kryptofix 2.2.2 - of 20.3 mg, potassium carbonate 4.3 mg, acetonitrile - 320 μl, water - 80 µl) in a heated reactor. The resulting mixture was dried in a current of dry nitrogen, and then the reactor was added a solution of mannose triflate in acetonitrile containing a certain quantity of water.

Next, the reactor was kept within 80 seconds when the external temperature of the heater is equal to 125°C, then took 0.6 ml, which was discarded to remove any traces of residual water from the tubes), and the residue was transferred into the vial with the product. The water content in the vial with the product was determined by Karl Fischer titration using 50 μl of the solution, and the radiochemical purity was determined using instant thin-layer chromatography (MTCH). Thin-layer chromatography was performed on plates of silica gel, designed to conduct for TLC, showing a mixture of 95% acetonitrile and 5% water, and then measured the relative abundance of [18F]-fluoride and 1,3,4,6-Tetra-O-acetyl-2-fluoro-β-D-mannopyranose (in all cases they were the only components) using MTSH.

Measurement of the ratio of binding water

To determine the reduction to which icesta water during the reaction with the triflate mannose spent two cold (non-radioactive) experience in which from the reactor was collected a certain amount of liquid before and after the introduction of the indicator. It is possible to determine the ratio of binding water in measured quantities of water.

Then perform the sequence of synthesis, a similar sequence of experiment introduction the radiotracer; 2 ml of water was passed through the cartridge, Waters Access PlusQMA (carbonate form), after which the cartridge was suirable with a solution of Kryptofix and carbonate in acetonitrile/water (Kryptofix 2.2.2 - of 20.3 mg, potassium carbonate 4.3 mg, acetonitrile - 320 μl, water - 80 µl) in a heated reactor. The resulting mixture was dried in a current of dry nitrogen, and then the reactor was added a solution of mannose triflate in acetonitrile containing a certain quantity of water.

After adding in the reactor solution of mannose triflate were selected to 0.6 ml solution, which was placed into the vial with the product. Further reaction of the introduction of the radiotracer was performed for 80 seconds when the external temperature of the heater is equal to 125°C, and then took the rest of the solution, which was placed in a separate vial with the product. The water content of each vial was determined by Karl Fischer titration using 50 μl of a solution.

The results of the experiments on the binding of water shown in Table 1, which shows the concentration of water present in the combined acetonitrile process is the.

Table 1
Before the beginning of the reaction (parts per million)After the reaction (parts per million)
786802
26032527
84337860

At low and medium concentrations of water significant water retention in the reaction with the triflate mannose was observed. However, at elevated concentrations in water was observed to reduce the water content of approximately 7%.

The results of the introduction of the radiotracer

To obtain the water content before the beginning of the reaction the introduction of the radiotracer measured the water content in each reaction the introduction of the radiotracer, which was then refined by introducing the coefficient of binding water by mannose triflate. The degree of radiochemical purity obtained for each concentration of water, are shown in Table 2 and shown in Figure 3.

Table 2
Parts per million of water prior to the reaction (calculated) Radiochemical purity, %
50694,6
70791,4
280397,6
385595,9
411496,7
577998,4
594394,1
798085,6
920673,6
1538285,0
4337585,5

These results confirm that the preferred water content is from 3000 to 6000 parts per million. If we exclude random result when 73,6% radiochemical purity, the results of the reaction are grouped around the radiochemical purity equal to 85%, even with a high moisture content.

1. The way to obtain secure fluorinated derivative of glucose, including interaction derived tetracylines with fluoride, characterized in that the reaction is carried out in a solvent containing water in excess of 1000 parts per Milli is h and less than 50,000 parts per million.

2. The method according to claim 1, in which protected fluorinated derivative of glucose represents 2-fluoro-1,3,4,6-Tetra-O-acetyl-D-glucose (tetraarylporphyrin or pFDG), and specified derivative tetracylines is a 1,3,4,6-Tetra-O-acetyl-2-O-trifloromethyl-β-D-mannopyranose (triplet tetracylines).

3. The method according to any of claim 1 or 2, wherein the solvent is selected from the group consisting of acetonitrile, dimethylformamide, dimethylsulfoxide, tetrahydrofuran, dioxane, 1,2-dimethoxyethane, sulfolane and N-methylpyrrolidinone.

4. The method according to claim 3, in which the solvent is a acetonitrile.

5. The method according to claim 1, in which the water content in the solvent is from about 1000 to 15,000 parts per million.

6. The method according to claim 5, in which the water content in the solvent is from about 2,000 to 7,000 parts per million.

7. The method according to claim 1, in which the water content in the solvent is from about 3000 to 6000 parts per million.

8. The method according to claim 7, which is carried out in solution.

9. The method according to claim 1, which is automated.

10. The method according to claim 1 in which the fluoride is a fluoride ion with potassium counterion, and fluoride added to the catalyst phase transfer, such as 4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo-[8,8,8]-hexacosane.

11. The method according to claim 1 for the preparation of radiator the one derived sugar.

12. The method according to claim 11, in which radiotolerance derived sugar is a sugar derived labeled with [18F].

13. The method of item 12, in which the specified derived sugar labeled with [18F], is a [18F]-pFDG.

14. The method of claim 1, also comprising one or more additional stages, carried out in any order:
1) removal of excess fluoride from the solution;
2) unprotect a protected fluorinated derivative of sugar from obtaining unsecured fluorinated derivative of sugar;
3) removing the organic solvent; and
4) introduction of unprotected fluorinated derivative of sugar in aqueous solution.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention refers to synthesis of [18F]fluororganic compounds ensured by reaction of [18F]fluoride and relevant halogenide or sulphonate with alcoholic vehicle of formula 1 where R1, R2 and R3 represent hydrogen atom or C1-C18 alkyl.

EFFECT: possibility for mild process with low reaction time and high yield.

21 cl, 2 tbl, 27 ex

FIELD: chemistry.

SUBSTANCE: invention claims derivatives of 1-α-halogen-2,2-difluoro-2-deoxy-D-ribofuranose of the general formula (I) in solid state, where R1 is benzoyl or ; R2 is hydrogen; and X is CI, Br or I; which can be applied as intermediates in stereoselective method of gemcitabine obtainment. In addition, invention claims stereoselective method of obtaining compounds of the general formula (I), including stages of: (i) recovery of 1-oxoribose of formula to obtain lactol of formula ; (ii) interaction of compound of formula (III) with halogen phosphate compound of formula in the presence of a base to obtain 1-phosphatefuranose derivative of formula ; and (iii) interaction of compound of formula (V) (also included in the claim) with halogen source, with further recrystallisation of obtained product; where R1, R2 and X are the same as indicated above while R3 is phenyl.

EFFECT: efficient method of obtaining derivatives of the abovementioned agent.

11 cl, 6 ex

FIELD: chemistry.

SUBSTANCE: in method of obtaining compound aminoalkyl glucosaminide 4-phosphate of formula , X represents , Y represents -O- or NH-; R1, R2 and R3, each is independently selected from hydrogen and saturated and unsaturated (C2-C24) aliphatic acyl groups; R8 represents -H or -PO3R11R11a, where R11a and R11a, each is independently -H or (C1-C4) aliphatic groups; R9 represents -H, -CH3 or -PO3R13aR14, where R13a and R14, each is independently selected from -H and (C1-C4) aliphatic groups, and where indices n, m, p, q each independently is a integer from 0 to 6 and r is independently integer from 2 to 10; R4 and R5 are independently selected from H and methyl; R6 and R7 are independently selected from H, OH, (C1-C4) oxyaliphatic groups -PO3H2, -OPO3H2, -SO3H, -OSO3H, -NR15R16, -SR15, -CN, -NO2, -CHO, -CO2R15, -CONR15R16, -PO3R15R16, -OPO3R15R16, -SO3R15 and -OSO3R15, where R15 and R16, each is independently selected from H and (C1-C4) aliphatic groups, where aliphatic groups are optionally substituted with aryl; and Z represents -O- or -S-; on condition that one of R8 and R9 represents phosphorus-containing group, but R8 and R9 cannot be simultaneously phosphorus-containing group, including: (a) selective 6-O- silylation of derivative of 2-amino-2-desoxy-β-D-glucopyranose of formula , where X represents O or S; and PG independently represent protecting group, which forms ester, ether or carbonate with oxygen atom of hydroxy group or which forms amide or carbamate with amino group nitrogen atom, respectively; by means of tri-substituted chlorosilane RaRbRcSi-Cl, where Ra, Rb and Rc are independently selected from group, consisting of C1-C6alkyl C3-C6cycloalkyl and optionally substituted phenyl, in presence of tertiary amin, which gives 6-silylated derivative; (b) selective acylation of 4-OH position of obtained 6-O-silylated derivative with 6-3-alkanoyloxyalcanoic acid or hydroxyl-protected (R)-3-hydroxyalkanoic acid presence of a carbodiimide reagent and catalytic 4-dimethylaminopyridine or 4-pyrrolidinopyridine to give a 4-O-acylated derivative; (c) selectively deprotecting the nitrogen protecting groups, sequentially or simultaneously and N,N-diacylating the resulting diamine with (R)-3-alkanoyloxyalkanoic acid or a hydroxy-protected (R)-3-hydroxyalkanoic acid in presence of peptide condensation reagent; (d) introducing a protecting phosphate group at 3-position with a chlorophosphate or phosphoramidite reagent to give a phosphotriester; and (e) simultaneous or sequential deprotecting phosphate, silyl, and remaining protecting groups.

EFFECT: method improvement.

11 cl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention concerns a variant of admixture extraction from composition containing extraneous matter and sucralose, which is used as a sweetener. One of the variants includes following stages: (a) first solvent extraction of the said composition containing sucralose and admixtures in the first solvent with the help of another solvent, at least partially immiscible, in order to remove admixtures to the said second solvent; (b) second solvent extraction of the said composition containing sucralose and admixtures in the first solvent with the help of the third solvent, at least partially immiscible, in order to transfer sucralose to the said third solvent; where stage (a) removes at least a part of admixtures to the second solvent; and stage (b) transports most of sucralose to the third solvent and detains most of admixtures in the first solvent.

EFFECT: efficient removal of admixtures from compositions.

34 cl, 4 tbl, 2 dwg, 2 ex

FIELD: chemistry.

SUBSTANCE: developed method of sucralose production using acyl-sucralose implies (a) adjustment of pH factor of specified supplied mixture to value ranged from 8.0 to 12.0 by alkali metal hydroxide addition; (b) buffer addition to specified base mixture in amount enough for specified pH factor stabilization within stated range over holding stage (c); (c) holding of specified base mixture at appropriate temperature over time period enough for effective transformation of specified acyl-sucralose compound into free sucralose; (d) reduction of specified pH factor of specified base mixture up to value from 4 to 8; (e) sucralose release from product of step (d) resulted thereby in released sucralose.

EFFECT: improved method of water deacylation procedure stabilization.

22 cl, 1 tbl, 1 ex

FIELD: organic chemistry, chemical technology.

SUBSTANCE: invention relates to an improved solid-phase method for synthesis of radioisotope indicators, in particular, for synthesis of compounds labeled with 18F that can be used as radioactive indicators for positron- emission tomography (PET). In particular, invention relates to a method for synthesis of indicator labeled with 18F that involves treatment of a precursor fixed on resin if the formula (I): SOLID CARRIER-LINKER-X-INDICATOR wherein X means a group promoting to nucleophilic substitution by a definite center of a fixed INDICATOR with 18F- ion for preparing a labeled indicator of the formula (II): 18F-INDICATOR; to compound of the formula (Ib):

and compound of the formula (Ih): ;

to radiopharmaceutical set of reagents for preparing indicator labeled with 18F for using in PET; to a cartridge for radiopharmaceutical set of reagents for preparing indicator labeled with 18F for using in positron-emission tomography.

EFFECT: improved method of synthesis.

13 cl, 1 sch, 3 ex

FIELD: pharmaceutical technology.

SUBSTANCE: invention relates to the improved sucralose formulation and a method for its crystallization. Method involves controlling pH value of solution in the range from about 5.5 to about 8.5 in the process of formation of sucralose crystals. Invention provides the development of the improved composition comprising crystalline sucralose and possessing the enhanced stability.

EFFECT: improved preparing method, improved properties of composition.

24 cl, 4 tbl, 4 ex

The invention relates to medicine, more specifically to radiopharmaceuticals for diagnostic purposes, and may find application in positron emission tomography

FIELD: medicine.

SUBSTANCE: invention refers to a single-stage method for producing a compound of formula I from the compound of formula ASC including interaction of a compound of formula ASC with the appropriate chlorinating agent in an organic solvent in the presence of a base, and segregating the compound of formula I from the reaction mixture, at that use of protective groups in the given method is not required.

EFFECT: new single-stage method for producing compound of formula I is developed.

7 cl, 1 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: disclosed are α- and β-crystalline forms of 5'-desoxy-N4-carbopentyloxy-5-fluorocytidine of formula (III) , their preparation method through crystallisation of the raw product from a suitable solvent and pharmaceutical compositions based on the said compounds, having anti-cancer activity. The solvent used when preparing the α-modification is an ester or a mixture of ester-containing solvents. The solvent used when preparing the β-modification is a mixture of water and alkanol or a mixture of tetrahydrofuran and diethyl ether or carbon tetrachloride.

EFFECT: obtaining compounds and pharmaceutical compositions based on the said compounds, having anticancer activity.

10 cl, 2 dwg, 7 ex

FIELD: chemistry.

SUBSTANCE: invention refers to sulphated oligosaccharides of the general formula X-[Y]n-Z-UR1, where X, Y and Z each are the same hexose monosaccharide fragment selected out of group including glucose, mannose, altrose, allose, talose, galactose, idose and gulose, adjoining monosaccharide fragments are bound in 1→2, 1→3, 1→4, and/or 1→6 pattern by glycoside bonds, and each carbon atom not binding X, Y and Z groups is bound by single bond with UR group, with exception for carbon atom in 1 position of Z monosaccharide, to which UR1 group is bound by single bond; where n is an integer within 0 to 6; U is O atom or NH; each R is independently C2-C6-alkenyl, benzyl, SO3M or H, where M is any pharmaceutically acceptable cation of alkali metal or organic amine, or R form N3 together with U; R1 is C1-C12alkyl, benzyl, PEG monomethyl ether or its derivative, C1-C12alkylazide, , or , in the form of ester, free acid, free base or hydrate. Also invention refers to pharmaceutical or veterinary composition based on claimed compounds, for disorder prevention or treatment for mammals in case of proliferate retinopathy, solid tumour and/or metastasis result, coagulation/thrombosis and/or virus infection of organism. Additionally invention refers to application of claimed compounds in medicine production for disorder prevention or treatment for mammals in case of proliferate retinopathy, solid tumour and/or metastasis result, coagulation/thrombosis and/or virus infection of organism.

EFFECT: increased efficiency of compound application in medicine.

11 cl, 3 tbl, 1 dwg, 16 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to a compound having general structural formula , in which n is equal to 1 or 5 and R6 is COOH or CH2OPO3H2, to a pharmaceutically acceptable salt of the said compound. The invention also pertains to a pharmaceutical composition based on the said compounds or their pharmaceutically acceptable salts, meant for inducing or boosting immunoreaction in a subject.

EFFECT: invention relates to a method of inducing or boosting immunoreaction in a subject, as well as to a method of relieving or essentially preventing an infectious disease, involving administration of an effective amount of the said compound to the subject.

7 cl, 5 ex

FIELD: medicine.

SUBSTANCE: invention concerns to a derivative of formulas (I) and to its application as an inhibitor of human sodium-dependent glucose transporter (SGLT) for treating the diseases associated with hyperglycemia: , where A1 is O or S; A2 is CH or N; R1 or R4 is a group of formula (S): , where R5, R6 are hydrogen, hydroxyl, halogen, alkyl, alkoxy-, alkylthio; Q is alkylene, alkenylene, alkylene-O- alkylen-S-, -O-alkylene-, -S-alkylene-, alkylene-O-alkylene, -alkylene-S-alkylene-; A is phenyl; R2, R3 - hydrogen; G is group of formulae: or E1 - hydrogen, fluorine, hydroxyl; E2 - hydrogen, fluorine, methyl, hydroxymethyl.

EFFECT: production of new inhibitors of human SGLT.

15 cl, 17 ex, 7 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to oligosaccharide substrates modified with a fluorescent group (see general structural formula given below ) which are soluble in water, for detecting cellulase complex enzymes on solid (agarised) media, where n (degree of polymerisation) equals 2-4, and the fluorescent group used is 2-(2'-hydroxyphenyl)-benzothiazole.

EFFECT: proposed are oligosaccharide substrates, modified with a fluorescent group, for detecting cellulase complex enzymes on solid media.

1 cl, 1 tbl, 2 dwg

FIELD: chemistry.

SUBSTANCE: invention refers to substrates of oligosaccharide class for detecting endo-glycosidehydrolase in presence of exo-acting ferments of general formula specified below, where X - oxygen atom or sulphur atom, and finite non-reducing sugar residual matter is connected to glyconic part of substrate molecule with thio-connection, at that n ≥ 0.

EFFECT: aglyconic part of the above substrates provides detection on solid (agarised) media and represents non-soluble fluorescent mark.

1 cl, 1 tbl

FIELD: medicine.

SUBSTANCE: invention is related to medicine, namely to method for production of liposome forms of antimicrobial preparations for treatment and prophylaxis of special danger infectious diseases. Method includes the following: solution of lipid and rifampicin shots in chloroform, preparation of gentamicin sulfate solution, creation of emulsion from chloroform solution of lipids with rifampicin, production of liposomes by evaporation with reversion of phases, removal of rifampicin that is non-included into membrane as well as gentamicin sulfate that is non-included inside vesicle by means of centrifugation, differing by the fact that in process of liposomes preparation by addition to chloroform mixture of rifampicin lipids in organic phase and increase of lipid load, high stability of emulsion and gel of "water in oil" type is achieved, which made it possible to build-in antibiotic into membrane. Universality of carrier is provided by neutral charge of liposome membrane. Used method makes it possible to apply one organic dissolvent with a certain temperature of boiling.

EFFECT: production of emulsion on rotor mixer eliminates toxic effect of titanium ions (titanium probe of ultrasonic disintegrator) onto membrane of liposomes and antibacterial preparations, reduces treatment time.

2 ex, 1 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to a cyclic bioisostere of purine system derivatives, with general structural formula given below , where R = , Li, Na or K, R1 = -H, -NH2, -Br, -Cl, -OH, -COOH; A = -N- for B=-N=, Z = -CH-; A = -CH= for B = -N=, Z = -CH-; A = -CH= for B = -N=, Z = -N=; A = -CH= for B = -CH=, Z - -CH=; A = -CH= for B = -CH=, Z = -N=, except compounds in which A = -CH= for B = -CH=, Z = -CH=, R= Li, Na or K and R1= -NH2 in the 5th position of the benzo[d]-3H-pyridazine-1,4-dione nucleus, and its pharmacologically acceptable salts, with normalising effect on intracellular processes.

EFFECT: obtaining compounds which can be used for normalising intracellular processes in therapy of disorders, caused by intracellular acidosis and/or oxygen deficiency and/or excess formation of free radicals and/or excess formation of free radical forms of oxygen and/or high thrombocyte aggregation and/or erythrocytes and/or adverse effects and/or nitrergic cell mechanism disorder.

17 cl, 14 tbl, 15 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to a cyclic bioisostere of purine system derivatives, with general structural formula given below , where R = , Li, Na or K, R1 = -H, -NH2, -Br, -Cl, -OH, -COOH; A = -N- for B=-N=, Z = -CH-; A = -CH= for B = -N=, Z = -CH-; A = -CH= for B = -N=, Z = -N=; A = -CH= for B = -CH=, Z - -CH=; A = -CH= for B = -CH=, Z = -N=, except compounds in which A = -CH= for B = -CH=, Z = -CH=, R= Li, Na or K and R1= -NH2 in the 5th position of the benzo[d]-3H-pyridazine-1,4-dione nucleus, and its pharmacologically acceptable salts, with normalising effect on intracellular processes.

EFFECT: obtaining compounds which can be used for normalising intracellular processes in therapy of disorders, caused by intracellular acidosis and/or oxygen deficiency and/or excess formation of free radicals and/or excess formation of free radical forms of oxygen and/or high thrombocyte aggregation and/or erythrocytes and/or adverse effects and/or nitrergic cell mechanism disorder.

17 cl, 14 tbl, 15 dwg

FIELD: organic chemistry, biochemistry, medicine.

SUBSTANCE: invention relates to phosphoramidates of nucleoside analogs comprising 2',3'-dideoxy-2',3'-didehydrothymidine 5'-phosphodimorpholidate of the formula (I) and phosphoramidates of 3'-azido-3'-deoxythymidine of the formula (II) and the formula (III) that inhibit activity in reproduction of human immunodeficiency virus (HIV). Compounds are resistant to effect of dephosphorylating enzymes and able to penetrate into cells and elicit the selective activity in inhibition of DNA biosynthesis catalyzed by HIV-reverse transcriptase.

EFFECT: valuable medicinal and biochemical properties of nucleoside analogs.

4 dwg, 1 tbl, 5 ex

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