KR102048050B1 - Synthetic method of metabolites of adb-fubinaca - Google Patents

Abstract

The present invention relates to a method for synthesizing a metabolite of ADB-fubinaka to develop an evaluation method whether ADB-fubinaka is taken, which is a novel hemp-like component. Through the method for synthesizing the metabolite of ADB-fubinaka according to an embodiment of the present invention, it is possible to synthesize the metabolite of ADB-fubinaka by a simple method.

Description

Method of synthesizing metabolite of ADB-Fubinaka {SYNTHETIC METHOD OF METABOLITES OF ADB-FUBINACA}

The present invention relates to a method for the simple synthesis of metabolites of ADB-fubinaka.

Since the late 2000s, new drugs such as synthetic hemp have increased rapidly, which has become a major international issue. The "new drug" is made by modifying the chemical structure of the existing hemp, which lacks scientific data and is spreading cheaply and rapidly through internet transactions. Synthetic cannabis is a hallucinogenic substance that can be inhaled by mixing with cannabis and mixing it with herbs, e-cigarettes, and incense. Synthetic hemp substances are frequently addicted and misused due to the stronger pharmacological effect than hemp, and have various adverse effects such as hypertension, paranoia and hallucinations. In many countries, efforts are being made to identify and regulate new hemp materials, but have difficulty coping with rapidly growing new materials. In particular, the information on the biological metabolite component of the synthetic hemp that must be confirmed in order to determine whether to take the synthetic hemp is less known, in most cases the standard is not available.

ADB-fubinaca is a new kind of hemp-like hemp that has recently emerged. It lacks information on its metabolism and lacks a commercially available standard, making it impossible to confirm whether or not ADB-fubinaka is taken. Development of a method for synthesizing ADB-Fubinaka's metabolites as it is essential to detect ADB-Fuvinaca's metabolites in biological samples such as urine and hair obtained from drug offenders This is necessary.

The technical problem to be achieved by the present invention is to provide a method for synthesizing a metabolite of ADB-fubinaka, a novel pseudo-hemp component.

However, the problem to be solved by the present invention is not limited to the above-mentioned problem, another task that is not mentioned will be clearly understood by those skilled in the art from the following description.

One embodiment of the present invention comprises the steps of (a) reacting a compound of formula (1) with a compound of formula R ' ₃ Si-X to prepare a compound of formula (2) having a hydroxy group protected; (b) preparing a compound of Formula 3 by halogenating the compound of Formula 2 and then reacting with a cyan compound; (c) reacting the compound of Formula 3 with 4-fluorobenzyl halide to prepare a compound of Formula 4; (d) hydrolyzing the compound of Formula 4 to prepare a compound of Formula 5; And (e) reacting the compound of formula 5 with a tert-leucine amide derivative to prepare a compound of formula (I).

Scheme 1

Compounds of the formula R ′ ₃ Si-X include trimethylsilyl halides; Triethylsilyl halide; Tri-n-propylsilyl halides; Tri-iso-propylsilyl halides; t-butyldimethylsilyl halide; t-butyldiphenylsilyl halide; And one or more of 2- (trimethylsilyl) ethoxymethyl halide;

In the above formulas, X is a halogen atom and R is a hydrogen atom or a hydroxy group.

Metabolites of ADB-fubinaka can be synthesized by a simple method through the synthesis method of the metabolite of ADB-fubinaka according to one embodiment of the present invention.

The metabolite of ADB-fubinaka can be synthesized at high yield through the method for synthesizing the metabolite of ADB-fubinaka according to one embodiment of the present invention.

Through the analysis of metabolites synthesized by the method of synthesizing the ADB-fubinaca metabolites according to an exemplary embodiment of the present invention, an analytical method in a sample such as urine or hair for confirming whether ADB-fubinaca is taken Can be.

In addition, synthetic hemps are expected to introduce new drugs that have changed the chemical structure of the existing synthetic hemp finely, and thus, may be applied to the study of synthetic hemps having similar chemical structure and similar biometabolism as ADB-fubinaka. Can be.

All technical terms used in the present invention, unless defined otherwise, are used in the meaning as commonly understood by those skilled in the art in the related field of the present invention. In addition, although a preferred method or sample is described herein, similar or equivalent things are included in the scope of the present invention. The contents of all publications incorporated herein by reference are incorporated by reference in their entirety.

Hereinafter, the present invention will be described in more detail.

One embodiment of the present invention comprises the steps of (a) reacting a compound of formula (1) with a compound of formula R ' ₃ Si-X to prepare a compound of formula (2) having a hydroxy group protected; (b) preparing a compound of Formula 3 by halogenating the compound of Formula 2 and then reacting with a cyan compound; (c) reacting the compound of Formula 3 with 4-fluorobenzyl halide to prepare a compound of Formula 4; (d) hydrolyzing the compound of Formula 4 to prepare a compound of Formula 5; And (e) reacting the compound of formula 5 with a tert-leucine amide derivative to prepare a compound of formula (I).

Scheme 1

Compounds of the formula R ′ ₃ Si-X include trimethylsilyl halides; Triethylsilyl halide; Tri-n-propylsilyl halides; Tri-iso-propylsilyl halides; t-butyldimethylsilyl halide; t-butyldiphenylsilyl halide; And one or more of 2- (trimethylsilyl) ethoxymethyl halide;

In the above formulas, X is a halogen atom and R is a hydrogen atom or a hydroxy group.

In the present specification, the compound of formula I wherein R is a hydrogen atom is compound I-1, and the compound of formula I wherein R is a hydroxy group is compound I-2.

The present invention can synthesize a compound of formula (I) which is a metabolite of ADB-fubinaka by a simple method through the synthesis method described above. In addition, according to an exemplary embodiment of the present invention, the ADB-fubinaca metabolite of Formula I may be obtained in high yield.

Hereinafter, a method for synthesizing the ADB-fubinaka metabolite of formula (I) according to an exemplary embodiment of the present invention will be described in more detail at each step.

Synthesis method of the ADB-fubinaka metabolite of formula (I) according to an exemplary embodiment of the present invention may be to prepare compounds by reacting the reactants in stoichiometric equivalents.

Step (a) according to an exemplary embodiment of the present invention is to react the compound of Formula 1 with a compound of Formula R ′ ₃ Si-X to form a silyl ether to protect the hydroxyl group of Formula 1. According to an exemplary embodiment of the present invention, the compound of the formula R ' ₃ Si-X is trimethylsilyl chloride, triethylsilyl chloride, tri-n-propylsilyl chloride, tri-iso-propylsilyl chloride, t-butyldimethyl At least one of silyl chloride and t-butyldiphenylsilyl chloride.

Specific examples of the step of protecting the hydroxy group of Chemical Formula 1 include the reaction of the compound of Chemical Formula 1 with t-butyldimethylsilyl chloride to silylate a hydroxy group to form t-butyldimethylsilyl ether. The step (a) may be performed for 12 hours to 24 hours at room temperature under DMF solvent conditions.

Step (b) according to an exemplary embodiment of the present invention is a halogenated compound of Formula 2, and then specifically, for example, a compound of Formula 2 using F ₂ , Cl ₂ , Br ₂ or I ₂ under strong base conditions. The hydrogen at position 3 may be a halide group (F, Cl, Br, or I), and then reacted with a cyanide compound to prepare a compound of Formula 3.

According to an exemplary embodiment of the present invention, the cyanide compound may be at least one of sodium cyanide, potassium cyanide and copper cyanide. Specifically, step (b) is performed by adding a compound of Formula 2, a halogen diatomic molecule, and a strong base such as sodium hydroxide or potassium hydroxide to, for example, dimethylformamide, and then performing the mixture at a room temperature for 0.5 to 2 hours. After the compound is halogenated, copper cyanide may be added, and the reaction may be performed at 150 ° C. to 200 ° C. for 5 minutes to 20 minutes to prepare the compound of Formula 3.

According to an exemplary embodiment of the present invention, step (c) may be performed under basic conditions. For example, the compound of Chemical Formula 3 and 4-fluorobenzyl halide may be reacted under strong base conditions to replace hydrogen with 4-fluorobenzyl group. Step (c) may be performed for 3 hours to 6 hours at 0 ° C to 25 ° C under THF solvent conditions.

According to an exemplary embodiment of the present invention, before the step (c) is performed to replace the position 1 hydrogen of the formula 3 with 4-fluorobenzyl group, step (b) is performed to cyanide position 3 hydrogen of the formula (2). The reason for substituting with the group is to prevent the substitution of hydrogen at position 3 of the higher reactivity with 4-fluorobenzyl. Therefore, the compound of formula (I), which is a metabolite of ADB-fubinaca, can be synthesized at high yield through the synthesis method according to one embodiment of the present invention.

Step (d) according to the exemplary embodiment of the present invention may be a step of removing (deprotecting) the protecting group of the compound of Formula 4 and hydrolyzing the cyanide group with a carboxylic acid group. According to an exemplary embodiment of the present invention, step (d) may be performed under basic conditions. For example, after the addition of a strong base potassium hydroxide or sodium hydroxide to the compound of formula 4 may be to perform the reaction for 1 hour to 3 hours at 90 ℃ to 110 ℃ temperature.

According to the method for synthesizing the ADB-fubinaca metabolite of Formula I, which is one embodiment of the present invention, hydrolysis of the protecting group and cyanide group of the compound of Formula 4 may occur simultaneously through step (d). In addition to synthesizing the metabolite of fubinaka, it is possible to increase the yield of ADB-fubinaka metabolite by preventing the loss of yield which can occur through several steps.

According to an exemplary embodiment of the present invention, the step (e) is a compound of formula 5 L-tert- leucine amide, D-tert- leucine amide or 2-amino-4-hydroxy-3,3-dimethylbutane And reacting with the amide. The step (e) may be performed for 24 hours or more under basic conditions and room temperature.

In conclusion, the present invention can synthesize the ADB-fubinaka metabolite of formula (I) by a simple method through the above-described synthesis method. In addition, through the analysis of the metabolites synthesized by the method of synthesizing ADB-fubinaca according to the present invention, an emotional technique for confirming whether or not to take ADB-fubinaca can be developed.

Compounds of formula (I) prepared according to one embodiment of the present invention are liquid chromatography-electrospray ionization-quadrupole / flight time mass spectrometry (LC-ESI-Q / TOF MS) for samples extracted from urine or hair. Can be used when analyzing the presence or absence of the metabolite of ADB-Fubinaka. For example, a chromatogram of a compound of formula (I), a synthesized ADB-fubinaka metabolite, compared to a chromatogram of a urine or hair sample, is present in the urine or hair sample, where a compound of formula (I) is an ADB-fubinaka metabolite. You can check whether you are taking ADB-fubinaca. Accordingly, it is possible to quickly and easily confirm whether or not a metabolite of ADB-fubinaka, a new like hemp component, exists in the urine or hair sample.

Hereinafter, the present invention will be described in detail with reference to Examples. However, embodiments according to the present invention can be modified in many different forms, the scope of the invention is not to be construed as limited to the embodiments described below. The embodiments of the present specification are provided to more completely explain the present invention to those skilled in the art.

Example  1 (Synthesis of Compound I-1)

Compound I-1 was synthesized under the conditions and methods described in Scheme 2 below.

Scheme 2

Compound of formula 2-1 (5-(( tert - Butyldimethylsilyl ) Oxy )-One H - Indazole Manufacture of

1.0 g (7.45 mmol) of 5-hydroxy- 1H -indazole (compound of Formula 1, Tokyo Chemical Industry, Inc.) and imidazole (Alpha Co., Ltd.) in 15 mL of dimethylformamide (DMF) at 25 ° C. 1.0 g (14.9 mmol) was dissolved, and 1.46 g (9.69 mmol) of t-butyldimethylsilyl chloride (TBSCl, Alfasa Co., Ltd.) was added, followed by reaction for 18 hours. Diluted with 15 mL of water, extracted with ethyl acetate (3 × 20 mL), washed with 20 mL of brine and dehydrated with sodium sulfate (Na ₂ SO ₄ ). Using a 15:85 volume ratio ethyl acetate and hexane mixed solution to give a white solid compound of the formula 2-1 by column chromatography (Yield: 99%).

The ¹ H-NMR and ¹³ C-NMR results for the obtained compound are as follows.

¹ H-NMR (600 MHz, CDCl ₃ ): δ 8.00 (s, 1H), 7.37 (d, J = 9.0 Hz, 1H), 7.14 (d, J = 2.4 Hz, 1H), 6.99 (dd, J = 9.0, 2.4 Hz, 1H), 1.02 (s, 9H), 0.22 (s, 6H); ¹³ C-NMR (150 MHz, CDCl ₃ ): δ 150.1, 136.4, 134.0, 123.9, 122.6, 110.5, 108.8, 25.9, 18.3, -4.3.

Compound of formula 3-1 (5-(( tert - Butyldimethylsilyl ) Oxy )-One H - Indazole -3- Carbon Nitrile Manufacture of

Dissolve 1.8 g (7.52 mmol) of the compound of Chemical Formula 2-1 obtained in 45 mL of dimethylformamide, add 3.68 g (14.5 mol) of iodine and 0.81 g (14.5 mmol) of potassium hydroxide (KOH), and then 25 After stirring for 1 hour at 0 ° C., the reaction solution was mixed with 90 mL of 10% aqueous sodium hydrogen sulfite (NaHSO ₃ ) solution. The mixture was extracted three times with 30 mL of ethyl acetate, and the organic layers were collected, washed three times with 30 mL of cold water, washed once more with brine, and then dehydrated with sodium sulfate. The solution was filtered and the solvent was blown off under reduced pressure distillation to prepare a yellow solid in which iodine was introduced into position 3 hydrogen of the compound of Formula 2-1. 2.7 g (7.25 mmol) of a yellow solid having iodine introduced into position 3 hydrogen of the compound of Formula 2-1 was dissolved in 25 mL dimethylformamide without purification, and 1.29 g (14.4 mmol) of copper cyanide (CuCN, TCI Co., Ltd.) was obtained. After the addition, the microwave reaction was performed at 185 ° C for 10 minutes. 30 mL of water and 30 mL of ethyl acetate were added to the reaction solution, and the insoluble precipitate copper cyanide was filtered through diatomaceous earth (celite). To remove dimethylformamide, the solution was washed three times with 30 mL of cold water, washed with brine and dehydrated with sodium sulfate. The solvent was removed by distillation under reduced pressure and purified by column chromatography using a mixture of ethyl acetate and hexane in a volume ratio of 8:92 to obtain a compound of formula 3-1 in the form of a yellow solid (Yield: 50%).

The ¹ H-NMR and ¹³ C-NMR results for the obtained compound are as follows.

¹ H-NMR (600 MHz, CDCl ₃ ): δ 7.63 (d, J = 9.0 Hz, 1H), 7.20 (d, J = 1.8 Hz, 1H), 7.13 (dd, J = 9.0, 2.4 Hz, 1H) , 1.02 (s, 9 H), 0.24 (s, 6 H); ¹³ C-NMR (150 MHz, CDCl ₃ ): δ 152.8, 136.2, 125.6, 124.8, 118.8, 114.0, 112.0, 107.0, 25.8, 18.3, -4.3.

Compound of formula 4-1 (5-(( tert - Butyldimethylsilyl ) Oxy ) -1- (4- Fluorobenzyl )-One H Preparation of Indazole-3-carbonitrile)

164.4 mg (1.46 mmol) of potassium t-butoxide (t-BuOK) was added to a solution of 334 mg (1.22 mmol) of the compound of Chemical Formula 3-1 obtained in 7 mL of anhydrous THF at 0 ° C. After stirring for 1 hour at this temperature, 0.18 mL (1.46 mmol) of 4-fluorobenzyl bromide (Alpha Co., Ltd.) was added dropwise, and the reaction was carried out for 4 hours at 25 ° C. The solvent was removed under reduced pressure, dissolved in 10 mL ethyl acetate, washed with water and brine and dehydrated with sodium sulfate. The reaction solution was condensed and separated by column chromatography purification (8:92 volume ratio of ethyl acetate and hexane mixed solution) to obtain a compound of formula 4-1 in the form of a yellow oil (Yield: 70%).

The ¹ H-NMR and ¹³ C-NMR results for the obtained compound are as follows.

¹ H-NMR (600 MHz, CDCl ₃ ): δ 7.28 (d, J = 9.0 Hz, 1H), 7.24-7.20 (m, 2H), 7.15 (d, J = 2.4 Hz, 1H), 7.04-6.99 ( m, 3H), 5.56 (s, 2H), 1.00 (s, 9H), 0.22 (s, 6H); ¹³ C-NMR (150 MHz, CDCl ₃ ): δ 162.8 (d, J = 245.5 Hz), 152.6, 135.6, 130.9 (d, J = 2.8 Hz), 129.4 (d, J = 7.2 Hz), 127.0, 123.9 , 117.2, 116.1 (d, J = 21.4 Hz), 113.9, 111.2, 107.4, 53.8, 25.7, 18.3, -4.3.

Compound of formula 5 (1- (4- Fluorobenzyl ) -5- Hydroxy -One H - Indazole Preparation of 3-carboxylic acid):

The solution of the compound of formula 4-1 obtained in 1.5 mL of 10 N sodium hydroxide (NaOH) aqueous solution was heated to 100 ° C. for 1.5 hours. After cooling to 25 ° C., the pH value of the solution was adjusted to 1 using 3N hydrogen chloride (HCl). The resulting precipitate was filtered and dried to yield the compound of formula 5 as a white solid (Yield: 78%).

The ¹ H-NMR and ¹³ C-NMR results for the obtained compound are as follows.

¹ H-NMR (600 MHz, CD ₃ OD): δ 7.46-7.42 (m, 2H), 7.30-7.27 (m, 2H), 7.06-6.98 (m, 3H), 5.63 (s, 2H); ¹³ C-NMR (150 MHz, CD ₃ OD): δ 165.8, 163.9 (d, J = 244.2 Hz), 155.3, 137.5, 135.2, 133.7, 130.5 (d, J = 8.6 Hz), 126.4, 120.1, 116.5 ( d, J = 21.4 Hz), 112.3, 105.1, 53.7.

Preparation of Compound I-1:

41 mg (0.14 mmol) of the compound of Chemical Formula 5 obtained above were dissolved in 1 mL of dimethylformamide, and 1.2 mg (0.22 mmol) of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDCI, TCI), 29.0 mg (0.2 mmol) of oxybenzotriazole (HOBt, Sigma Aldrich), 0.13 mL (0.72 mmol) of N, N-diisopropylethylamine (DIPEA, Alfasa) and 28.0 mg of L-tert-leucine amide (0.2 mmol) and stirred for 24 hours. 5 mL of water was added and extracted with 5 mL of ethyl acetate, and the separated water layer was further extracted three times with 5 mL of ethyl acetate. The combined organic layers were collected, washed three times with 5 mL of cold water, and sodium sulfate was used to remove water remaining in the organic solvent. The reaction solution condensed under reduced pressure was purified by column chromatography using methanol and dichloromethane as a mixed solution, to obtain Compound I-1 as a white solid (Yield: 78%).

The ¹ H-NMR and ¹³ C-NMR results for the obtained compound are as follows.

¹ H-NMR (600 MHz, CD ₃ OD): δ 7.84 (d, J = 9.6 Hz, 1H), 7.51 (d, J = 2.4 Hz, 1H), 7.39 (d, J = 9.6 Hz, 1H), 7.27 (q, J = 5.4 Hz, 2H), 7.03 (t, J = 9.0 Hz, 2H), 6.98 (dd, J = 9.6, 2.4 Hz, 1H), 5.61 (s, 2H), 4.52 (d, J = 9.6 Hz, 1H), 1.09 (s, 9H); ¹³ C-NMR (150 MHz, CD ₃ OD): δ 175.2, 164.4, 163.8 (d, J = 244.2 Hz), 155.0, 137.8, 136.8, 133.9, 130.4 (d, J = 8.7 Hz), 125.4, 120.0, 116.4 (d, J = 21.6 Hz), 112.1, 105.0, 61.1, 53.6, 35.6, 27.2.

Compound I-1 prepared in Example 1 was analyzed by LC-ESI-Q / TOF method, and the detection time and m / z values of the fragment ions are shown in Table 2 below.

LC-ESI-Q / TOF analysis conditions used in the present invention are as follows.

The model name of the equipment is HR-Q / TOF system.

Ultra-High-Performance Liquid Chromatography (MSHP)

* UHPLC terms

Column: Bruker Intensity solo C18 (100 x 2.1 mm, 1.8 μm)

Mobile phase:

Solvent A-aqueous solution containing 1% methanol, 0.01% formic acid, 5 mM ammonium formate

Solvent B-methanol with 0.01% formic acid and 5 mM ammonium formate

Gradient profile is shown in Table 1 below

Time (min) Flow (ml / min) Solvent A (%) Solvent B (%) 0 0.2 96.0 4.0 0.1 0.2 96.0 4.0 1.0 0.2 82.0 18.0 2.5 0.2 50.0 50.0 14.0 0.4 0.1 99.9 16.0 0.5 0.1 99.9

Injection volume: 2 μL, column temperature: 40 ° C * MS / MS conditions

Ionization Method: Electrospray Ionization (ESI)

Detection method: positive ion, full scan mode

* ESI condition

Drying gas flow rate (DGF): 8 L / min

Drying gas temperature (DGT): 200 ° C

Nebulizing gas flow: 8 L / min

Collision energy: 6.0 eV

Capillary voltage: (+) 3500 V

Detection time (minutes) m / z Compound i-1 7.9 399.1839 Fragment Ions of Compound I-1 - 382, 354, 269

Through Example 1 it can be seen that through the synthesis method of the ADB-fubinaka metabolite according to an embodiment of the present invention it is possible to synthesize ADB-fubinaka metabolite with a high yield. In addition, it can be seen that through the analysis of the synthesized metabolites, an emotional technique for confirming whether ADB-fubinaca is taken or not can be developed.

Claims (6)

(a) reacting a compound of formula 1 with a compound of formula R ′ ₃ Si-X to prepare a compound of formula 2 wherein the hydroxy group is protected;
(b) preparing a compound of Formula 3 by halogenating the compound of Formula 2 and then reacting with a cyan compound;
(c) reacting the compound of Formula 3 with 4-fluorobenzyl halide to prepare a compound of Formula 4;
(d) hydrolyzing the compound of Formula 4 to prepare a compound of Formula 5; And
(e) reacting the compound of formula 5 with a tert-leucine amide derivative to prepare a compound of formula (I);
Scheme 1

Compounds of the formula R ′ ₃ Si-X include trimethylsilyl halides; Triethylsilyl halide; Tri-n-propylsilyl halides; Tri-iso-propylsilyl halides; t-butyldimethylsilyl halide; t-butyldiphenylsilyl halide; And one of 2- (trimethylsilyl) ethoxymethyl halide;
In the above formulas, X is a halogen atom and R is a hydrogen atom or a hydroxyl group.
The method of claim 1,
Compounds of the formula R ′ ₃ Si-X include trimethylsilyl chloride, triethylsilyl chloride, tri-n-propylsilyl chloride, tri-iso-propylsilyl chloride, t-butyldimethylsilyl chloride and t-butyldiphenylsilyl A method for synthesizing an ADB-fubinaka metabolite of formula (I), which is at least one of chlorides.
The method of claim 1,
The cyanide compound is at least one of sodium cyanide, potassium cyanide and copper cyanide ADB-Fubinaka metabolite synthesis method of formula (I).
The method of claim 1,
The step (c) is carried out under basic conditions ADB-Fubinaka metabolite synthesis method of formula (I).
The method of claim 1,
(D) step is carried out under basic conditions ADB-Fubinaka metabolite synthesis method of formula (I).
The method of claim 1,
The tert-leucine amide derivatives are L-tert-leucine amide, D-tert-leucine amide or 2-amino-4-hydroxy-3,3-dimethylbutanamide. .