CN111018858A

CN111018858A - Heterocyclic derivative and preparation method and application thereof

Info

Publication number: CN111018858A
Application number: CN201811176278.9A
Authority: CN
Inventors: 昌军; 汪美玲; 金琳; 张贺延昊; 牛彤
Original assignee: Fudan University
Current assignee: Fudan University
Priority date: 2018-10-10
Filing date: 2018-10-10
Publication date: 2020-04-17
Anticipated expiration: 2038-10-10
Also published as: CN111018858B

Abstract

The invention belongs to the technical field of pharmaceutical chemistry and medicine, and relates to a novel heterocyclic derivative, a preparation method and application thereof. The invention provides a novel heterocyclic derivative shown as a formula (1), wherein R represents a substituted or unsubstituted five-membered or six-membered heterocyclic ring. The invention also provides a preparation method of the heterocyclic derivative and adenosine A_2AReceptor antagonistic activity and use as antitumor drug.

Description

Heterocyclic derivative and preparation method and application thereof

Technical Field

The invention belongs to the technical field of pharmaceutical chemistry and medicine, and particularly relates to a novel heterocyclic derivative, and a preparation method and application thereof.

Background

The prior art discloses that the tumor immunotherapy is the fourth major tumor treatment method following surgical treatment, radiotherapy and chemotherapy drug treatment, and has made rapid progress in recent years, leading to a major breakthrough in the field of tumor therapy. The american clinical oncology society of 2016 and 2017 has rated "immunotherapy" as an annual progression of antitumor studies for two consecutive years. It is reported that the tumor immunotherapy is a therapeutic strategy for stimulating and enhancing the immune function of the body by various means, and finally achieving the purpose of controlling and killing tumor cells through the body's own immune system. According to Lerink Partners' prediction, the global tumor immunotherapy drug market size will reach $ 167 billion by 2020, and the tumor immunotherapy drug market size will achieve more than 2-fold increase to $ 361 billion by 2025. Currently, the field and research focus in which tumor immunotherapy is more successful has mainly focused on immune checkpoint inhibitors.

Since 2011 Ipilimumab (CTLA-4 inhibitor, bristol, precious) was approved by the FDA in the united states for marketing as the first immunotherapeutic agent for melanoma, research in this area has progressed at a surprising rate. Nivolumab (PD-1 inhibitor, bosch, bai shi gui bao, 2014), Pembrolizumab (PD-1 inhibitor, merck, 2014) and Atezolizumab (PD-L1 inhibitor, roche, 2016) were each sequentially approved by the U.S. FDA for marketing. In addition against novel immune checkpoints such as indoleamine 2, 3-dioxygenase-1 (IDO 1), adenosine A_2AReceptor (A)_2AR), T cell immunoglobulin, mucin-3 (TIM-3), and the like have also been the focus of research.

Adenosine A_2AR antagonists have been studied previously as therapeutic parkinson agents: istradefylline, although approved in Japan, is terminated by the United states FDA; the clinical phase III of Preladenant was terminated; vipadenant clinical phase II was terminated. In recent years, the research on tumor immunotherapy discovers A_2AThe R antagonist SCH58261 significantly inhibited the metastatic spread of CD73 positive tumors (B16F 10 melanoma) by increasing NK cell function, and SCH58261 or ZM241365 was found to increase the activity of anti-PD 1 monoclonal antibody or anti-CTLA 4 monoclonal antibody when administered in combination. Gov website, Nowa and Palobioofama, Inc. conducted clinical phase I/Ib trials for PBF-509 alone or in combination with the PD-1 inhibitor PDR001 for the treatment of non-small cell lung cancer, and phase I/Ib trials for CPI-444 alone or in combination with the PD-1 inhibitor Atezolizumab, by Gentek and Corvus, Inc. conducted clinical phase I/Ib trials for the treatment of solid tumors. In addition, the company Asricon and Heptaretherapeutics were also prepared to initiate HTL-1071 alone or in combination with the PD-L1 inhibitor DurvalumabClinical phase I test of non-small cell lung cancer by medication. Redox therapeutics was purchased by CAR-T Juno corporation 2016 to obtain its small molecule drug Vipadenant ready for combination therapy with CAR-T therapy.

Based on the current state of the art, the inventors of the present application intend to provide novel heterocyclic derivatives, and preparation methods and uses thereof.

Disclosure of Invention

The present invention aims to provide a novel heterocyclic derivative based on the current state of the art.

The invention also aims to provide a preparation method and application of the novel heterocyclic derivative.

The invention provides a heterocyclic derivative shown as the following formula (1):

（1）

in the formula, R represents a substituted or unsubstituted five-membered or six-membered heterocyclic ring.

The invention also provides a preparation method of the heterocyclic derivative, which comprises the following specific steps:

step a: carrying out reflux reaction on 5-bromo-4-chloropyrimidine-2-amine (2) and heterocycle substituted carbohydrazide compounds (3) in the absence of a solvent or in a solvent to generate 5-bromo-pyrimidine-2-amine derivatives (4);

step b: 5-bromo-pyrimidin-2-amine derivatives (4), andN,Obis-trisilylacetamide (cas: 10416-59-8), hexamethyldisilazane (cas: 999-97-3), cyclizing to form 8-bromo- [1,2,4] under the condition of no solvent or in the presence of solvent by refluxing reaction]Triazolo [1,5-c]Pyrimidine-5-amine compounds (5);

step c: the 8-bromo- [1,2,4] triazolo [1,5-c ] pyrimidine-5-amine compound (5) and 1-methyl-6-oxo-1, 6-dihydropyridine-3-boronic acid pinacol ester are subjected to reflux reaction in a solvent under the action of a catalyst under the alkaline condition to generate the corresponding heterocyclic compound (1).

In the present invention, the solvent used in step a may be any solvent as long as the solvent itself is inert in the reaction and does not inhibit the reaction; the solvent includes halogenated hydrocarbon solvents such as dichloromethane, chloroform, 1, 2-dichloroethane, etc.; aromatic hydrocarbon solvents such as benzene and toluene, etc.; aprotic solvents such as acetone, acetonitrile, N-dimethylformamide, N-methyl-2-pyrrolidone, dimethyl sulfoxide, sulfolane, cyclobutanesulfoxide, and hexamethylenephosphoric triamide; ester solvents such as ethyl acetate and methyl acetate, etc.; ether solvents such as tetrahydrofuran, diethyl ether and 1, 4-dioxane; organic base solvents such as pyridine, picoline, etc.; protic solvents such as methanol, n-butanol, etc.; or mixtures of these solvents. The reaction temperature can be between 80 ℃ and 120 ℃, and preferably 100 ℃ to 120 ℃; the molar ratio of the starting materials may be any, but 2:1 is preferably used.

In the present invention, the solvent used in step b may be any solvent as long as the solvent itself is inert in the reaction and does not inhibit the reaction; the solvent includes halogenated hydrocarbon solvents such as dichloromethane, chloroform, 1, 2-dichloroethane, etc.; aromatic hydrocarbon solvents such as benzene and toluene, etc.; aprotic solvents such as acetone, acetonitrile, N-dimethylformamide, N-methyl-2-pyrrolidone, dimethyl sulfoxide, sulfolane, cyclobutanesulfoxide, and hexamethylenephosphoric triamide; ester solvents such as ethyl acetate and methyl acetate, etc.; ether solvents such as tetrahydrofuran, diethyl ether and 1, 4-dioxane; organic base solvents such as pyridine, picoline, etc.; protic solvents such as methanol, n-butanol, etc.; or mixtures of these solvents. The reaction temperature can be between 80 ℃ and 120 ℃, and preferably 100 ℃ to 120 ℃; the molar ratio of the starting materials may be arbitrary, and 1:1:1 is preferably used.

In the present invention, the base in step c may be an organic base such as triethylamine, pyridine, N-diisopropylethylamine, 4-dimethylaminopyridine, 1, 8-diazabicyclo [5.4.0] -7-undecene, 1,2,2,6, 6-pentamethylpiperidine, etc., or an inorganic base such as sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, etc.; the solvent used in the preparation method may be any solvent as long as the solvent itself is inert in the reaction and does not inhibit the reaction; the solvent includes halogenated hydrocarbon solvents such as dichloromethane, chloroform, 1, 2-dichloroethane, etc.; aromatic hydrocarbon solvents such as benzene and toluene, etc.; aprotic solvents such as acetone, acetonitrile, N-dimethylformamide, N-methyl-2-pyrrolidone, dimethyl sulfoxide, sulfolane, cyclobutanesulfoxide, and hexamethylenephosphoric triamide; ester solvents such as ethyl acetate and methyl acetate, etc.; ether solvents such as tetrahydrofuran, diethyl ether and 1, 4-dioxane; organic base solvents such as pyridine, picoline, etc.; or a mixture of these solvents, the preferred solvent being water, 1, 4-dioxane. The reaction temperature can be between 80 ℃ and 120 ℃, and preferably 80 ℃ to 100 ℃; the molar ratio of the starting materials may be any, and 1:1 is preferably used. The catalyst may be a palladium-based catalyst, and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium is preferably used.

The heterocyclic compound can be used for preparing medicines, in particular to tumor immunotherapy medicines.

In the present invention, adenosine A is performed in vitro on the compound represented by the general formula (1)_2AThe results of the activity test and the in vivo anti-tumor effect of the receptor show that the compound has adenosine A_2AReceptor antagonistic activity and antitumor action, and thus can be used for preparing a medicament containing the above heterocyclic compound as an active ingredient.

The novel heterocyclic compound can be prepared into a pharmaceutical composition containing a safe and effective amount of the novel heterocyclic compound and a medicinal carrier and various preparations.

In the present invention, the "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. The safe and effective amount is determined according to the age, condition, course of treatment, etc. of the subject.

By "pharmaceutically acceptable carrier" is meant: one or more compatible solid or liquid fillers or gel substances which are suitable for human use and must be of sufficient purity and sufficiently low toxicity.

By "compatible" is meant that the components of the composition are capable of intermixing with and between the compounds of the present invention without significantly diminishing the efficacy of the compounds.

Examples of pharmaceutically acceptable carrier moieties are sugars such as glucose, sucrose, lactose, and the like; starches, such as corn starch, potato starch, and the like; cellulose and its derivatives such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.; gelatin, talc, solid lubricants, such as stearic acid, magnesium stearate; calcium sulfate, vegetable oils such as soybean oil, sesame oil, peanut oil, olive oil, etc.; polyhydric alcohols such as propylene glycol, glycerin, mannitol, sorbitol, and the like; emulsifiers, such as Tween; wetting agents, such as sodium lauryl sulfate; coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

For the sake of understanding, the present invention will be described in detail below by way of specific examples. It is specifically noted that the specific examples are given for illustrative purposes only, and it will be apparent to those skilled in the art that various modifications and variations can be made in the present invention within the scope of the present invention based on the descriptions herein.

Detailed Description

Example 1: synthesis of heterocyclic Compound (1 a)

Step a: 5-bromo-4-chloropyrimidin-2-amine (2) (370.27 mg, 2.94 mmol) and 2-furancarbohydrazide (3 a) (300 mg, 1.44 mmol) were dissolved in 10 mL of n-butanol, reacted at 120 ℃ under reflux under nitrogen for 12 hours, concentrated under reduced pressure to remove the solvent, and subjected to silica gel column Chromatography (CH)₂Cl₂MeOH =10: 1) to give N' - (2-amino-5-bromopincidin-4-yl) furan-2-carbohydrazide (4 a),¹H NMR (400 MHz, DMSO) δ (ppm): 8.10 (s, 1H), 7.22(s, 1H), 7.23-7.22 (d,J= 3.6 Hz, 1H), 6.62-6.61 (t,J= 1.6 Hz, 1H)。ESI-MS：298 [M+1]⁺。

step b: in the reaction of 5-bromo-pyrimidin-2-amine derivative (4 a) (150 mg, 503.19 mmol) withN,OHexamethyldisilazane (cas: 999-97-3, 1.5mL) was added to bis (trimethylsilyl) acetamide (cas: 10416-59-8, 1.5mL), and the mixture was refluxed at 120 ℃ for 12 hours under nitrogen atmosphere, concentrated under reduced pressure to remove the solvent, and subjected to silica gel column Chromatography (CH)₂Cl₂MeOH =10: 1) to give 8-bromo-2- (furan-2-yl) - [1,2,4]triazolo[1,5-c]pyrimidin-5-amine（5a）；¹H NMR (400 MHz, DMSO) δ (ppm): 8.16 (s, 2H), 8.04 (s, 1H), 7.98-7.96 (d,J= 3.2, 1H), 7.27-7.26 (d,J= 3.2 Hz, 1H), 6.74-6.73 ( d,J= 1.6Hz, 1H)。ESI-MS：280 [M+1]⁺。

Step c: 8-bromo- [1,2,4]Triazolo [1,5-c]Pyrimidine-5-amine compound (5 a) (110 mg, 392.74mmol), 1-methyl-6-oxo-1, 6-dihydropyridine-3-boronic acid pinacol ester (92.33 mg, 392.74mmol, 92.33uL), potassium carbonate (108.56 mg, 785.48 mmol), dissolved in 1, 4-dioxane (3.00 mL) and water (300mL), reacted at 90 ℃ under reflux under nitrogen for 12 hours, concentrated under reduced pressure to remove the solvent, and chromatographed on silica gel Column (CH) under reduced pressure₂Cl₂MeOH =10: 1), yielding compound 1a (11.80 mg, yield 9.75%).

5-(5-amino-2-(furan-2-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-8-yl)-1-methylpyridin-2(1H)-one (1a):¹H NMR (400 MHz, DMSO) δ (ppm): 8.43 (d, J =2.3 Hz, 1H), 8.06 (dd, J = 8.8, 2.6 Hz, 1H), 8.02 (s, 1H), 7.72 (s, 1H), 7.26(d, J = 3.5 Hz, 1H), 6.66 – 6.61 (m, 2H), 3.66 (s, 3H).¹³C NMR (400 MHz,MeOD) δ (ppm)：160.90, 158.18, 154.31, 152.31, 147.05, 146.18, 139.38, 136.86,120.22, 113.73, 113.02, 110.23, 101.42, 38.55。ESI-MS：309 [M+1]⁺。

Example 2: synthesis of heterocyclic Compound (1 b-i)

8-bromo- [1,2,4]Triazolo [1,5-c]Pyrimidin-5-amine compound (5)b-i) (400 mmol), 1-methyl-6-oxo-1, 6-dihydropyridine-3-boronic acid pinacol ester (400 mmol), potassium carbonate (800 mmol), dissolved in 1, 4-dioxane (4.00 mL) and water (400 mL), reacted under reflux at 100 ℃ for 10 hours under nitrogen protection, concentrated under reduced pressure to remove the solvent, and chromatographed over silica gel Column (CH)₂Cl₂MeOH =10: 1) to give compounds 1 b-i.

5-(5-amino-2-(5-methylfuran-2-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-8-yl)-1-methylpyridin-2(1H)-one (1b).¹H NMR (400 MHz, DMSO) δ (ppm): 8.50 (s,1H), 8.15 – 7.96 (m, 4H), 7.18 (d,J= 1.2 Hz, 1H), 6.53 (d,J= 9.7 Hz, 1H),6.36 (d,J= 0.9 Hz, 1H), 3.54 (s, 3H), 2.40 (s, 3H).¹³C NMR (400 MHz, DMSO)δ (ppm)：161.02, 156.02, 154.42, 151.41, 146.93, 143.91), 142.44, 138.79 ,137.40, 119.04, 113.84,111.56, 108.46, 107.90, 37.29, 13.50. ESI-MS：323 [M+1]⁺。

5-(5-amino-2-(pyridin-2-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-8-yl)-1-methylpyridin-2(1H)-one (1c).¹H NMR (400 MHz, DMSO) δ (ppm): 8.77 (d,J=4.2 Hz, 1H), 8.57 (d,J= 1.7 Hz, 1H), 8.37 (d,J= 7.8 Hz, 1H), 8.21 – 8.00(m, 5H), 7.58 (dd,J= 7.0, 5.2 Hz, 1H), 6.55 (d,J= 9.5 Hz, 1H), 3.56 (s,3H).¹³C NMR (400 MHz, DMSO) δ (ppm)：162.59, 161.05, 151.65, 149.72, 148.77,147.10, 142.11, 138.81, 137.46, 125.27, 123.41, 119.06, 111.57, 108.36,37.34. ESI-MS：320 [M+1]⁺。

5-(5-amino-2-(thiazol-2-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-8-yl)-1-methylpyridin-2(1H)-one (1d).¹H NMR (400 MHz, DMSO) δ (ppm): δ 8.46 (d,J=1.7 Hz, 1H), 8.20 – 8.09 (m, 5H), 8.04 (d,J= 2.9 Hz, 1H), 6.55 (d,J= 9.5Hz, 1H), 3.54 (s,3H).¹³C NMR (400 MHz, DMSO) δ (ppm)：161.02, 158.12, 157.84,151.70, 147.07, 144.65, 142.94, 138.92, 137.46, 123.54, 119.07, 111.39 ,108.34, 37.29. ESI-MS：326 [M+1]⁺。

5-(5-amino-2-(1-methyl-1H-pyrazol-5-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-8-yl)-1-methylpyridin-2(1H)-one (1e).¹H NMR (400 MHz, DMSO) δ(ppm): 8.57 (d,J= 1.6 Hz, 1H), 8.21 – 8.05 (m, 4H), 7.59 (d,J= 0.6 Hz,1H), 7.00 (d,J= 1.0 Hz, 1H), 6.53 (d,J= 9.5 Hz, 1H), 4.32 (s, 3H), 3.54(s, 3H)。¹³C NMR (400 MHz, DMSO) δ (ppm)：161.00, 155.70, 151.06, 146.84,142.47, 138.52, 138.19, 137.48, 133.08, 119.14, 111.41, 108.29, 108.03,37.32. ESI-MS：323 [M+1]⁺。

5-(5-amino-2-(1-methyl-1H-pyrazol-3-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-8-yl)-1-methylpyridin-2(1H)-one (1f).¹H NMR (400 MHz, DMSO) δ(ppm): 8.52 (d,J= 0.4 Hz, 1H), 8.14 (d,J= 12.7 Hz, 2H), 7.99 (s, 2H),7.87 (s, 1H), 6.89 (s, 1H), 6.53 (d,J= 9.4 Hz, 1H), 3.96 (s, 3H), 3.54 (s,3H).¹³C NMR (400 MHz, DMSO) δ (ppm)：161.05, 158.95, 151.33, 146.94, 142.51,142.02, 138.88, 137.37, 132.26 , 119.02, 111.70, 108.09, 106.26, 37.31. ESI-MS：323 [M+1]⁺。

5-(5-amino-2-(6-methylpyridin-2-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-8-yl)-1-methylpyridin-2(1H)-one (1g).¹H NMR (400 MHz, DMSO) δ (ppm): 8.57 (d,J= 1.9 Hz, 1H), 8.20-7.89 (m, 5H), 7.91 (t,J= 7.6 Hz, 1H), 7.43 (d,J=7.8 Hz, 1H), 6.55 (d,J= 9.5 Hz, 1H), 3.56 (s, 3H), 2.58 (s, 3H).¹³C NMR(400 MHz, DMSO) δ (ppm)：162.84, 161.05, 158.36, 151.55, 148.39, 147.15,142.21, 138.84, 137.46, 124.67, 120.82, 119.07, 111.60, 108.28, 37.33, 24.06.ESI-MS：334 [M+1]⁺。

5-(5-amino-2-(5-methylthiazol-2-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-8-yl)-1-methylpyridin-2(1H)-one (1h).¹H NMR (400 MHz, DMSO) δ (ppm): 8.46 (d,J= 2.0 Hz, 1H), 8.18 (s, 1H), 8.16 – 8.09 (m, 3H), 7.79 (s, 1H), 6.55 (d,J= 9.6 Hz, 1H), 3.53 (s, 3H), 2.56 (s, 3H).¹³C NMR (400MHz, DMSO) δ (ppm)：1601.01 , 155.90, 154.56, 152.29, 1467.03, 144.78, 142.73, 138.88, 137.42,129.46, 127.24, 119.06, 108.26, 99.53, 37.30, 11.65. ESI-MS：340 [M+1]⁺。

5-(5-amino-2-(4-methylthiazol-2-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-8-yl)-1-methylpyridin-2(1H)-one (1i).¹H NMR (400 MHz, DMSO) δ (ppm): δ 8.44(d,J= 2.5 Hz, 1H), 8.20 – 8.08 (m, 4H), 7.58 (s, 1H), 6.55 (d,J= 9.4 Hz,1H), 3.54 (s, 3H).¹³C NMR (400 MHz, DMSO) δ (ppm)：161.04, 158.14,156.93,154.21, 151.68, 147.09, 142.95, 138.99, 137.45, 119.07, 117.91, 111.42,37.30, 16.81. ESI-MS：340 [M+1]⁺。

Example 3: compounds 1a-i adenosine receptor A in vitro_2AR and A₃Affinity assay for R

Detection of Compound-to-A separately Using radioligand competitive binding assay_2AR and A₃Affinity of R receptor: a in HeLa cells_2AR and 2³H]ZM241385 specific binding, A in HeLa cells₃R and 2³H]-NECA specific binding. A membrane protein solution (60 mg/tube containing ADU 2 mg/mL) and a radioactive ligand of 1 nM 0.2 nM [ mu ] m ] was added to each tube³H]ZM241385 et al, non-specific tubes added with 10 mM NECA/R-PIA at final concentration, compound tubes added with compounds at different concentrations, shaking and mixing, incubation at 37 ℃ for 30 min, reaction termination in water bath, negative pressure suction filtration using GF/C glass fiber filter paper to separate free ligand and bound ligand, washing 3 times with pre-cooled 50 mM Tris-HCl, about 4 mL each time. The film was removed, inverted on a tray (i.e., grid side up), and baked for 3 min until dry. The machine is cleaned in the same way as before. The dried small round filter was placed in an EP tube (pressed to the bottom of the tube) in order and 540 mL of scintillation fluid was added. Is carried out in a Beckman LS-6500 model multifunctional liquid scintillation counter³H]And (6) counting. Two double pipes are made at each joint point, and an average value is taken.

TABLE 1 affinity of Compounds 1a-i for adenosine receptorsKi (nM)

Compound (I)	A_2AR	A₃R
			1a	5.582	8790
1b	447.8	2964
			1c	62.44	>10000
1d	84.62	>10000
			1e	797.2	>10000
1f	3427	>10000
			1g	3767	>10000
1h	43.84	2934
			1i	1032	>10000

Example 4: compounds 1a to A_2AAntagonism of R

A₁R and A₃R can inhibit adenylate cyclase and inhibit cyclic adenosine monophosphate (cAMP) generation; and A is_2AR and A_2BR activates adenylate cyclase, inducing an increase in cAMP levels. 5' -N-ethyllcarboxamido-adenosines (NECA) is a stable analogue of adenosine and is not readily degraded by natural enzymes and is commonly used in place of adenosine. The experiment shows that the A can be expressed endogenously_2BIn HEK293 cells of R, 1 mM Compound 1a completely inhibited cAMP production in cells stimulated by NECA, indicating that it has A_2AR antagonism.

Example 5: research on antitumor effect of compound 1a in mice

B16 murine melanoma model: b16 murine melanoma cells were suspended in PBS and dilutions (2.0X 10)⁵Cell/cell) were inoculated in the hindpaw of six-week-old C57BL/6 male mice, and compound 1a (1 mg/kg) was administered intraperitoneally (i.p) (1 time per week for 4 times) and a blank control group of 6 mice each. Tumor volumes were measured every two days for a total of 28 days for efficacy evaluation. The experimental result shows that the tumor volume of the blank control group at day 28 is 552 +/-83 mm³And the tumor volume of the administration group at day 28 is 281 +/-34 mm³，（P<0.01), which shows that the compound 1a can obviously delay the growth of melanin tumor of B16 mouse after being treated;

initial anti-tumor growth mechanism study: further day 28, CD25 was detected by flow cytometry⁺Foxp3⁺Regulatory T cells are in CD4⁺The ratio of T cells in the test results showed that the ratio of the blank control group was 11.3. + -. 2.1%, and the ratio of the administered group was 5.2. + -. 0.9%, (P)<0.01), indicating that Compound 1a antagonizes A_2AR, inhibiting the differentiation of T cells into regulatory T cells, and promoting anti-tumor immunity.

Claims

1. Heterocyclic derivatives represented by formula (1):

（1）

2. A process for the preparation of heterocyclic derivatives according to claim 1, characterized in that it comprises the steps of:

step a: carrying out reflux reaction on 5-bromo-4-chloropyrimidine-2-amine (2) and heterocycle substituted carbohydrazide compounds (3) under the solvent-free condition or in a solvent to generate 5-bromo-pyrimidine-2-amine derivatives (4);

step b: 5-bromo-pyrimidin-2-amine derivatives (4), andN,Obis-trimethylsilyl acetamide and hexamethyldisilazane under solvent-free conditions or in a solvent to form 8-bromo- [1,2,4]Triazolo [1,5-c]Pyrimidine-5-amine compounds (5);

3. The method of claim 2, wherein the solvent in step a is N, N-dimethylformamide, toluene, dimethylsulfoxide or N-butanol.

4. The method of claim 2, wherein the solvent in step a is n-butanol.

5. The method of claim 2, wherein the solvent in step b is N, N-dimethylformamide, toluene, dimethylsulfoxide or N-butanol.

6. The method of claim 2, wherein the solvent in step b is n-butanol.

7. The method of claim 2, wherein the catalyst in step c is a palladium-based catalyst.

8. The method of claim 2, wherein the catalyst in step c is [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium.

9. Use of heterocyclic derivatives according to claim 1 for the preparation of adenosine a_2AUse in a formulation of receptor antagonistic activity.

10. Use of the heterocyclic derivative according to claim 1 for the preparation of an anti-tumor medicament.