CN118546097A - A 4,6-diaryl-2-substituted pyrimidineguanidine compound and its preparation method and application - Google Patents

Abstract

A 4,6-diaryl-2-substituted pyrimidineguanidine compound and its preparation method and application, relating to the field of organic synthesis, the compound has a novel structure, has a certain inhibitory effect on histone demethylase, and belongs to a new type of inhibitor structure. In addition, the preparation method of the compound is simple, easy to operate, has low requirements for equipment, and is suitable for large-scale industrial production in the later stage. The compound undergoes structural modification and in-depth pharmacological research, and has good application prospects in the preparation of histone demethylase inhibitors.

Description

A 4,6-diaryl-2-substituted pyrimidineguanidine compound and its preparation method and application

Technical Field

The present invention relates to the field of organic synthesis, and in particular to a 4,6-diaryl-2-substituted pyrimidineguanidine compound and a preparation method and application thereof.

Background Art

Among the pre-transcriptional histone modifications, lysine methylation has been the most widely studied. Histone lysine methylation modification is one of the causes of epigenetic changes. The enzymes that regulate lysine methylation are mainly divided into two categories: histone lysine methyltransferases (HKMTs) and histone lysine demethylases (HKDMs). HKMTs are responsible for transferring methyl groups from methyl donors to histone lysine to complete the methylation modification. The function of HKDMs is mainly to remove methyl groups from histone lysine to complete the demethylation process. The two have opposite biological functions and jointly regulate the methylation state of histone lysine. The misregulation of histone lysine demethylases (HKDMs) is closely related to the occurrence and development of many diseases, such as senile diseases and tumors. HKDMs are also related to the resistance of tumor drugs. Therefore, HKDMs have become an important target for the development of new anti-tumor drugs and are receiving more and more attention.

There are two types of histone lysine demethylases (HKDMs) discovered so far: one is lysine specific demethylases (LSDs), also known as FAD (flavin adenine dinucleotide)-dependent amine oxidases, including LSD1 and LSD2. The other is JmjC (Jumonji domain containing protein) histone demethylases, which are a family of proteins containing Jumonji domains in their molecules. Unlike LSD1, it can catalyze the demethylation of trimethylated lysine. The N-terminus and C-terminus of its family members contain a small Jumonji characteristic domain, called JmjN and JmjC, respectively. Among them, the JmjN domain is related to transcriptional regulation, while JmjC is a component of the active center of the JmjC family enzymes. Some JmjCs containing JmjC domains that have been discovered so far show different substrate specificity, and their demethylation catalytic process depends on the participation of Fe (II) and α-ketoglutarate (2-OG).

Histone demethylase is an emerging drug target. It has been less than 20 years since the first type of histone demethylase LSD family was discovered, and the second type JmjC family was discovered even later. Currently, research on the JmjC family is still in the development stage. Although various types of JmjC inhibitors have been discovered in recent years, most of them are derived from other targets, and inevitably there are selectivity issues and clinical efficacy issues. JmjC histone demethylase has gradually become an important target for epigenetic research and has received more and more attention. The development of new JmjC inhibitors is expected to provide new ideas for anti-tumor quality.

Summary of the invention

The purpose of the present invention is to provide a 4,6-diaryl-2-substituted pyrimidineguanidine compound and a preparation method and application thereof. The structure of the compound is different from the JmjC histone demethylase inhibitor reported in previous studies and belongs to a new type of inhibitor structure. By structurally modifying the compound of the present invention and conducting in-depth pharmacological studies, it is expected to develop a new histone demethylase inhibitor.

The embodiment of the present invention is achieved as follows:

A 4,6-diaryl-2-substituted pyrimidineguanidine compound having the structural formula

wherein _R1 is selected from hydrogen, C1-C9 alkyl, C1-C9 substituted alkyl, C2-C9 alkenyl, C2-C9 substituted alkenyl, C2-C9 alkynyl, C2-C9 substituted alkynyl, C1-C9 alkoxy, C1-C9 substituted alkoxy, C5-C10 cycloalkyl, C5-C10 heterocycloalkyl, C6-C10 aryl, or C6-C10 substituted aryl;

_R2 and _R3 may be the same or different, and are independently selected from C6-C10 aryl groups or C6-C10 substituted aryl groups.

A method for preparing the above-mentioned 4,6-diaryl-2-substituted pyrimidineguanidine compound comprises:

Compound I is subjected to a substitution reaction with a substituted amine to obtain compound II;

Compound II is hydrolyzed under the action of an alkaline reagent to obtain a 4,6-diaryl-2-substituted pyrimidineguanidine compound;

Wherein, the structural formula of compound I is The structural formula of the substituted amine is R ₄ NH ₂ , and the structural formula of compound II is

When R ₁ is hydrogen, the substituted amine is p-nitrobenzylamine, and when compound II is hydrolyzed, R ₄ is hydrolyzed to hydrogen; when R ₁ is not hydrogen, R ₄ and R ₁ are the same group.

A use of the above 4,6-diaryl-2-substituted pyrimidineguanidine compounds in the preparation of histone demethylase inhibitors.

The beneficial effects of the embodiments of the present invention are:

The embodiment of the present invention provides a 4,6-diaryl-2-substituted pyrimidine guanidine compound and its preparation method and application. The compound has a novel structure, has a certain inhibitory effect on histone demethylase, and belongs to a new type of inhibitor structure. In addition, the preparation method of the compound is simple, easy to operate, has low requirements for equipment, and is suitable for large-scale industrial production in the later stage. The compound undergoes structural modification and in-depth pharmacological research, and has good application prospects in the preparation of histone demethylase inhibitors.

DETAILED DESCRIPTION

In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

The following is a detailed description of a 4,6-diaryl-2-substituted pyrimidineguanidine compound according to an embodiment of the present invention, a preparation method thereof, and an application thereof.

A 4,6-diaryl-2-substituted pyrimidineguanidine compound having the structural formula

wherein _R1 is selected from hydrogen, C1-C9 alkyl, C1-C9 substituted alkyl, C2-C9 alkenyl, C2-C9 substituted alkenyl, C2-C9 alkynyl, C2-C9 substituted alkynyl, C1-C9 alkoxy, C1-C9 substituted alkoxy, C5-C10 cycloalkyl, C5-C10 heterocycloalkyl, C6-C10 aryl, or C6-C10 substituted aryl;

_R2 and _R3 may be the same or different, and are independently selected from C6-C10 aryl groups or C6-C10 substituted aryl groups.

Preferably, _R1 is selected from hydrogen, C1-C6 alkyl, C1-C6 substituted alkyl, C2-C6 alkenyl, C2-C6 substituted alkenyl, C2-C6 alkynyl, C2-C6 substituted alkynyl, C1-C6 alkoxy, C1-C6 substituted alkoxy, C5-C8 cycloalkyl, C5-C8 heterocycloalkyl, C6-C8 aryl, or C6-C8 substituted aryl;

_R2 and _R3 may be the same or different, and are independently selected from C6-C8 aryl groups or C6-C8 substituted aryl groups.

Among them, the alkyl group can be a straight chain alkyl group or a branched chain alkyl group, including but not limited to methyl, ethyl, n-propyl, isopropyl, tert-butyl, etc. Substituted alkyl refers to a group in which at least one hydrogen atom in the alkyl group is replaced by a halogen, hydroxyl, alkoxy or aryl group. Alkenyl can be a straight chain alkenyl group or a branched chain alkenyl group, including but not limited to vinyl, propenyl, allyl, butenyl, etc., and the number of double bonds can be single or multiple. Substituted alkenyl refers to a group in which at least one hydrogen atom in the alkenyl group is replaced by a halogen, hydroxyl, alkoxy or aryl group. Alkyl can be a straight chain alkynyl group or a branched chain alkynyl group, including but not limited to ethynyl, propynyl, propargyl, butynyl, etc. Substituted alkynyl refers to a group in which at least one hydrogen atom in the alkynyl group is replaced by a halogen, hydroxyl, alkoxy or aryl group. Alkoxy includes but is not limited to methoxy, ethoxy, propoxy, butoxy, etc. Substituted alkoxy refers to a group in which at least one hydrogen atom in the alkyl group is replaced by a halogen, hydroxyl, alkoxy or aryl group, including but not limited to 2-methoxyethoxy, 3-methoxypropoxy, etc. Cycloalkyl includes but is not limited to cyclopropyl, cyclobutyl, cyclohexyl, methylcyclohexyl, etc. Heterocycloalkyl refers to a group in which at least one carbon atom in the cycloalkyl is replaced by oxygen, nitrogen, sulfur, etc. Aryl includes but is not limited to phenyl, pyridyl, naphthyl, etc. Substituted aryl refers to a group in which at least one hydrogen atom in the aryl is replaced by halogen, hydroxyl, alkoxy or aryl.

The embodiment of the present invention also provides a method for preparing the above-mentioned 4,6-diaryl-2-substituted pyrimidineguanidine compound, which comprises:

Compound I is subjected to a substitution reaction with a substituted amine to obtain compound II;

Compound II is hydrolyzed under the action of an alkaline reagent to obtain a 4,6-diaryl-2-substituted pyrimidineguanidine compound;

Wherein, the structural formula of compound I is The structural formula of the substituted amine is R ₄ NH ₂ , and the structural formula of compound II is

When R ₁ is hydrogen, the substituted amine is p-nitrobenzylamine, and when compound II is hydrolyzed, R ₄ is hydrolyzed to hydrogen; when R ₁ is not hydrogen, R ₄ and R ₁ are the same group.

Furthermore, the molar ratio of compound I to substituted amine is 1:2 to 2.5. Since the raw material of substituted amine is easier to obtain, the amount of substituted amine is excessive to ensure that compound I can fully react and improve the product yield.

Optionally, the reaction of compound I and substituted amine is carried out under the catalysis of EDCI and DIPEA, and the molar ratio of compound I, EDCI and DIPEA is 1:1.8-2.2:1.8-2.2. EDCI can activate the C=S bond, so that the substituted amine can better perform nucleophilic reaction and attack the carbon atom of the C=S bond to obtain a guanidine structure.

Further, compound I is obtained by reacting compound III with benzoyl isothiocyanate, and the structural formula of compound III is

The amino group of compound III attacks the carbon atom of isothiocyanate to obtain compound I. The molar ratio of compound III to benzoyl isothiocyanate is 1:3-5, the reaction temperature is 70-80°C, and the reaction time is 5-8h. Under the above reaction conditions, compound III can be fully reacted to improve the product yield.

Furthermore, compound III is obtained by condensation reaction of compound IV and guanidine hydrochloride under base catalysis, and the structural formula of compound IV is

Compound IV and guanidine undergo 1,4 addition condensation reaction under base catalysis to obtain compound III. The dosage of compound IV and guanidine hydrochloride is 1:2.5-3; the reaction temperature is 70-80°C, and the reaction time is 7-10h. Under the above reaction conditions, compound IV can be fully converted into compound III.

In addition, the structure of compound IV is relatively common and can be directly purchased through commercial channels. It can also be synthesized through an aldol condensation reaction using aromatic formaldehyde and aromatic ethyl ketone as raw materials.

The embodiment of the present invention also provides a use of the above 4,6-diaryl-2-substituted pyrimidineguanidine compounds in the preparation of histone demethylase inhibitors.

Furthermore, the histone demethylase inhibitor includes 4,6-diaryl-2-substituted pyrimidineguanidine compounds, salts thereof or derivatives thereof.

When preparing the salt of 4,6-diaryl-2-substituted pyrimidineguanidine compounds, it is only necessary to react the 4,6-diaryl-2-substituted pyrimidineguanidine compounds with the corresponding acid. For example, when preparing the hydrochloride thereof, it is only necessary to react the 4,6-diaryl-2-substituted pyrimidineguanidine compounds with concentrated hydrochloric acid.

When preparing derivatives of 4,6-diaryl-2-substituted pyrimidineguanidine compounds, conventional reaction types are used to carry out substitution, oxidation-reduction, addition reaction, etc. For example, 4,6-diaryl-2-substituted pyrimidineguanidine compounds are reacted with carboxylic acid to prepare amide compounds.

The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

Example 1

This embodiment provides a 2-(4-(4-methoxyphenyl))-6-phenylpyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine and its hydrochloride (denoted as A01), and the preparation method thereof is as follows:

S1. Preparation of Compound IV

Benzaldehyde (2.12 g, 20 mmol) was placed in a 500 mL round-bottom flask and dissolved in 100 mL of methanol. A potassium hydroxide aqueous solution (150 mmol, 6.0 g of potassium hydroxide dissolved in 54 mL of water) and 4-methoxyacetophenone (3.00 g, 20 mmol) were added under ice bath, stirred for 0.5 h, then the ice bath was removed, and the reaction was stopped after stirring at room temperature for 10 h. After the reaction was completed, a large amount of precipitate was precipitated, which was filtered and washed with 200 mL of a mixed solvent (methanol: water = 1:1). After the filter residue was dried, intermediate 1 was obtained, which could be used directly in the next step without further purification, with a yield of 82%.

S2. Preparation of Compound III

Take a 250mL round-bottom flask, dissolve guanidine hydrochloride (3.58g, 37.5mmol) in 100mL ethanol, add sodium hydroxide (1.5g, 37.5mmol), and stir for 0.5h to free guanidine. Then put (E)-1-(4-methoxyphenyl)-3-phenylprop-2-en-1-one (compound IV, 3.57g, 15mmol) in another 500mL round-bottom flask and dissolve it in 100mL ethanol. Pour the ethanol solution of free guanidine into the ethanol solution of intermediate 1 and reflux at 75℃ for 8h. After the reaction is completed, evaporate the solvent under reduced pressure and purify by column chromatography (dichloromethane: ethyl acetate = 40:1) to obtain compound III with a yield of 24%.

S3. Preparation of Compound I

Place 4-(4-methoxyphenyl)-6-phenylpyrimidine-2-amine (Compound III, 0.83 g, 3 mmol) in a 100 mL round-bottom flask and dissolve it with 40 mL acetonitrile, and add benzoyl isothiocyanate (1.96 g, 12 mmol). Reflux at 75°C for 5 h. After the reaction is complete, a large amount of precipitate will precipitate. Filter and wash with 100 mL ethanol. After drying the filter residue, Compound I is obtained, which can be directly used in the next step without further purification, with a yield of 77%.

S4. Preparation of Compound II

4-Aminotetrahydropyran (0.10 g, 1 mmol) was placed in a 25 mL round-bottom flask and dissolved with 15 mL of dichloromethane, then DIPEA (0.13 g, 1 mmol) was added and stirred at room temperature for 0.5 h. N-((4-(4-methoxyphenyl)-6-phenylpyrimidin-2-yl)aminomethylthio)benzamide (Compound I, 0.22 g, 0.5 mmol) and EDCI (0.19 g, 1 mmol) were then added to the reaction solution. After stirring at room temperature for 10 h, the reaction was stopped, 75 mL of dichloromethane was added to the reaction solution to dilute, and the mixture was washed with water three times (3×30 mL) and saturated brine three times (3×30 mL). The organic phase was dried over anhydrous sodium sulfate, dried under reduced pressure, and purified by column chromatography to obtain Compound II with a yield of 87%.

S5. Preparation of 2-(4-(4-methoxyphenyl))-6-phenylpyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine

(E)-N-(N-(4-(4-methoxyphenyl))-6-phenylpyrimidin-2-yl)-N'-(tetrahydro-2H-pyran-4-yl)carbamoyl)benzamide (Compound II, 0.22 g, 0.44 mmol) was placed in a 50 mL round-bottom flask and dissolved with 10 mL of a mixed solvent (tetrahydrofuran: methanol = 1:1). Sodium hydroxide aqueous solution (40 mmol, 1.5 g sodium hydroxide dissolved in 3.5 mL water) was added to the reaction solution and refluxed at 70°C for 3 h. After the reaction was completed, the reaction solution was poured into 100 mL of water, and a precipitate was precipitated. The precipitate was filtered and washed with a large amount of water, and then recrystallized with methanol to obtain intermediate 5. Yield: 83%.

S6. Preparation of 2-(4-(4-methoxyphenyl))-6-phenylpyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride

Take a 25mL round-bottom flask, add all the 2-(4-(4-methoxyphenyl))-6-phenylpyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine (Intermediate 5) and 10mL methanol. Add 5-10 drops of concentrated hydrochloric acid to the solution and stir at room temperature for 48h. A large amount of precipitate is precipitated, filtered and washed with 50mL methanol, and the filter residue is collected to obtain product A01. Yield: 93%; white solid.

The characterization of compound A01 is as follows:

Mp: 186-187℃; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.20 (s, 1H), 9.87 (d, J = 7.9Hz, 1H),

9.05(s,2H),8.32-8.07(m,5H,aromatic),7.75-7.38(m,3H,aromatic),7.12(d,J＝8.7Hz,2H,aromatic),4.02-3.88(overlap,3H ₁₃ CNMR ⁽ 100MHz,DMSO-d ₆ ,ppm)δ:165.3,165.2,162.8,157.9,153.6,136.0,132.2,129.7,129.5,128.0,127.9,114.9,107.9,65.8,56.0,47.8,32.4.HRMS-ESI:m/z calculated for C ₂₃ H ₂₆ N ₅ O ₂ [M+H] ⁺ 404.2081, found 404.2075.

Embodiments 2 to 68

Examples 2 to 68 provide a series of 4,6-diaryl-2-substituted pyrimidineguanidine compounds and salts thereof (respectively referred to as A02 to A68). The preparation methods thereof are referred to Example 1. The structural formulas of compounds A02 to A68 are shown in Table 1.

Table 1. Structural formula of 4,6-diaryl-2-substituted pyrimidineguanidine hydrochloride

Compound A02: 2-(4-(2-hydroxyphenyl))-6-phenylpyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 14%; Mp: 173-174°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.42 (s, 2H), 9.73 (s, 1H), 8.94 (s, 2H), 8.32 (s, 1H, aromatic), 8.20-8.12 (m, 2H, aromatic), 8.09 (dd, J＝7.9, 1.1 Hz, 1H, aromatic), 7.68-7.54 (m, 3H, aromatic), 7.47-7.39 (m, 1H, aromatic), 7.08 (d, J＝8 .2Hz,1H,aromatic),7.05-6.94(m,1H,aromatic),4.02-3.81(m,3H,tetrahydropyranyl),3.45(overlap,2H,tetrahydropyranyl),2.05-1.89(m,2H,tetrahydropyranyl),1.63-1.46(m,2H,tetrahydropyranyl); ¹³ CNMR (100MHz, DMSO-d ₆ , ppm) δ: 165.7, 165.0, 158.5, 156.9, 153.4, 136.1, 133.7, 132.2, 130.1, 129.6, 127.8, 120.5, 120.0, 118.2, 110.2, 65.8, 47. 9,32.4.HRMS-ESI:m/z calculated for C ₂₂ H ₂₄ N ₅ O ₂ [M+H] ⁺ 390.1925, found 390.1929.

Compound A03: 2-(4-(3-nitrophenyl))-6-phenylpyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 17%; Mp: 213-215°C; ¹ H NMR (600 MHz, DMSO-d ₆ , ppm) δ: 11.30 (s, 1H), 9.68 (d, J = 7.9 Hz, 1H), 9.02 (s, 2H), 8.70 (d, J = 7.8 Hz, 1H, aromatic), 8.45 (s, 1H, aromatic), 8.40 (dd, J = 8.2, 2.5 Hz, 1H, aromatic), 8.23 (d, J = 7.5 Hz, 2H, aromatic), 7.86 (t, J = 8.0 Hz, 1H, aromatic), 7.67 -7.53(m,3H,aromatic),4.01-3.94(m,1H,tetrahydropyranyl),3.93-3.85(m,2H,tetrahydropyranyl),3.49-3.41(m,2H,tetrahydropyranyl),2.05-1.94(m,2H,tetrahydropyranyl),1.66-1.48(m ,2H,tetrahydropyranyl); ¹³ C NMR (150MHz, DMSO-d ₆ , ppm) δ: 165.7, 162.9, 157.5, 152.9, 148.4, 137.0, 135.1, 133.9, 132.1, 130.7, 129.1, 127.6, 126.1, 121.8, 108.9, 65.4, 47. 5,32.0.HRMS-ESI:m/z calculated for C ₂₂ H ₂₃ N ₆ O ₃ [M+H] ⁺ 419.1826, found 419.1820.

Compound A04: 2-(4-(4-bromophenyl)-6-(4-methoxyphenyl)pyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 12%; Mp: 275-276°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.26 (s, 1H), 9.79 (s, 1H), 9.04 (s, 2H) ),8.25-8.09(m,5H,aromatic),7.77(d,J＝8.4Hz,2H,aromatic),7.10(d,J＝8.8Hz,2H,aromatic),4.00-3.87(m,3H,tetrahydropyranyl ¹³ _C NMR(100MHz,DMSO-d ₆ ,ppm)δ:164.9,163.5,162.4,157.5,153.1,134.7,132.0,129.3,129.3,127.4,125.5,114.4,107.3,65.4,55.6,47.4,32.0.HRMS-ES I :m/z calculated for C ₂₂ H ₂₅ BrN ₅ O ₂ [M+H] ⁺ 482.1186, found 482.1185.

Compound A05: 2-(4,6-di(4-methoxyphenyl))pyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 18%; Mp: 280-282°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.12 (s, 1H), 9.89 (s, 1H), 9.05 (s, 2H) ),8.34-8.01(overlap,5H,aromatic),7.11(d,J＝7.8Hz,4H,aromatic),4.03-3.88(overlap,3H,tetrahydropyranyl),3.86(s,6H,2OCH ₃ ),3.50- 3.41(overlap,2H,tetrahydropyranyl),2.10-1.89(m,2H,tetrahydropyranyl),1.66-1.48(m,2H,tetrahydropyranyl). ¹³ CNMR(100MHz,DMSO-d ₆ ,ppm)δ:164.3,162.2,157.3,153.2,129.1,127.7,114.4,106.5,65.4,55.5,47.4,40.1,39.9,39.7,39.5,39.3,39.1,38.9,32.0.HRMS-ESI: m/z calculated for C ₂₄ H ₂₈ N ₅ O ₃ [M+H] ⁺ 434.2187, found 434.2182.

Compound A06: 2-(4,6-bis(4-bromophenyl))pyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 16%; Mp: 285°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.29 (s, 1H), 9.69 (d, J = 7.6Hz, 1H), 9.01 ( s,2H),8.31(s,1H,aromatic),8.14(d,J＝8.5Hz,4H,aromatic),7.77(d,J＝8.6Hz,4H,aromatic),4.02-3.79(overlap,3H, ¹³ CNMR (100MHz, DMSO-d ₆ ,ppm)δ:164.6,157.9,153.4,134.8,132.5,129.8,126.3,108.6,65.9,47.9,32.4.HRMS-ESI:m/z calculated for C ₂₂ H ₂₂ N ₅ OBr ⁸¹ Br[M+H] ⁺ 532.0165, found 532.0158.

Compound A07: 2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

Light yellow solid; total yield 21%; Mp: 207-208°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.24 (s, 1H), 9.73 (d, J = 7.3Hz, 1H) ,9.04(s,2H),8.92(s,1H,aromatic),8.73(d,J＝7.8Hz,1H,aromatic),8.45(d,J＝8.1Hz,1H,aromatic),8.39(s,1H ,aromatic),8.24(d,J＝8.7Hz,2H,aromatic),7.90(t,J＝8.0Hz,1H,aromatic),7.14(d,J＝8.8Hz,2H,aromatic),4.06-3.94( m,3H,tetrahydropyranyl),3.92(s,3H,OCH ₃ ),3.55-3.47(m,2H,tetrahydropyranyl),2.15-1.93(m,2H,tetrahydropyranyl),1.71-1.57(m,2H,tetrahydropyranyl); ¹³ C NMR (100MHz, DMSO-d ₆ ,ppm)δ HRMS-ESI: m/z calculated for C ₂₃ H ₂₅ N ₆ O ₄ [M+H] ⁺ 449.1932, found 449.1924.

Compound A08: 1-(2-chlorobenzyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 19%; Mp: 247-248°C; ¹ H NMR (600 MHz, DMSO-d ₆ ,ppm)δ:11.47(s,1H),9.96(s,1H),9.10(s,2H),8.91(s,1H,aromatic),8.58(d,J＝7.6Hz,1H,aromatic),8.42-8.27(m,2H,aromatic),8.02(d,J＝8.1Hz,2H,aromatic),7.78(t ,J＝8.0Hz,1H,aromatic),7.70(d,J＝6.8Hz,1H,aromatic),7.57(d,J＝7.5Hz,1H,aromatic),7.51-7.38(m,2H,aromatic),6.94(d,J＝8.4Hz,2H,aromatic),4.79(d,J＝5.3Hz,2H,N-CH ₂ ),3 .83(s,3H,OCH ₃ ^; 0,125.9,121.9,114.3,108.0,55.5,43.3.HRMS-ESI:m/z calculated for _{C 25} H ₂₂ N ₆ O ₃ Cl[M+H] ⁺ 489.1436,found 489.1426.

Compound A09: 1-(allyloxy)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 22%; Mp: 179°C; ¹ H NMR (600MHz, MeOD-d ₁ , ppm) δ: 8.83 (s, 1H, aromatic), 8.51 (s, 1H, aromatic), 8.35 (d ,J＝6.3Hz,1H,aromatic),8.13-7.97(m,3H,aromatic),7.76(s,1H,aromatic),7.02(d,J＝5.2Hz,2H,aromatic),6.18(m,1H ,C-CH＝C),5.56(d,J＝17.1Hz,1H,C＝CH _A ),5.48(d,J＝10.1Hz,1H,C＝CH _B ),4.60(d,J＝5.9Hz ,2H,O-CH ₂ -C),3.88(s,3H,OCH ₃ ); ¹³ C NMR (150MHz, MeOD-d ₁ , ppm) δ: 166.1, 163.2, 163.0, 157.3, 155.6, 148.7, 137.2, 133.1, 131.2, 130.2, 129.1, 127.1, 125.5, 121.6, 121.6, 114.2, 10 7.8,78.1 ,54.8.HRMS-ESI:m/z calculated for C ₂₁ H ₂₁ N ₆ O ₄ [M+H] ⁺ 421.1619,found421.1619.

Compound A10: 1-(4-hydroxyphenyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 24%; Mp: 190°C; ¹ H NMR (400 MHz, DMSO-d ₆ , ppm) δ: 11.32 (s, 1H), 9.97-9.67 (overlap, 2H), 8.96-8.70 (overlap, 3H), 8.50 (d, J = 7.8 Hz, 1H, aromatic), 8.34 (dd, J = 8.1, 1.6 Hz, 1H, aromatic), 8.25 (s, 1H, aromatic), 7.84-7.69 (m, 3H, aromatic), 7.36 (d, J = 8.4 Hz, 2H, aromatic), 6.93-6.74 (m, 4H, aromatic), 4.49 (d, J = 4.2 Hz, 2H, N-CH ₂ -), 3.82 (s, 3H, OCH ₃ ); ¹³ CNMR(100MHz,DMSO-d ₆ ,ppm)δ:164.5,162.5,162.2,157.7,157.3,153.4,148.3,136.9,133.5,130.4,130.2,129.1,126.7,125.9,125.3,121.8,115 .8,114.2,107.6,55.4,45.3.HRMS-ESI:m/z calculated for C ₂₅ H ₂₃ N ₆ O ₄ [M+H] ⁺ 471.1775, found 471.1767.

Compound A11: 1-(4-fluorobenzyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 20%; Mp: 172-173°C; ¹ H NMR (400 MHz, DMSO-d ₆ ,ppm)δ:11.40(s,1H),9.92(s,1H),9.05(s,2H),8.86(s,1H,aromatic),8.54(d,J＝7.8Hz,1H,aromatic),8.35(dd,J＝8.2,2.2Hz,1H,aromatic),8.27(s,1H,aromatic),7.91(d,J ＝8.5Hz,2H,aromatic),7.75(t,J＝8.0Hz,1H,aromatic),7.61(dd,J＝8.3,5.5Hz,2H,aromatic),7.29(t,J＝8.6Hz,2H,aromatic),6.87(d,J＝8.4Hz,2H,aromatic),4.67(d,J＝5.2Hz,2H,N-CH ₂ ), 3.81 (s, 3H, OCH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 164.8, 163.2 (d, ¹ J _CF = 244.2Hz), 162.4, 162.3, 157.3, 153.8, 148.3, 136.9, 133.5, 132.0 (d, ⁴ J _CF = 3.2Hz), 130.6, (d, ³ J _CF = 8.5Hz), 130.4, 129.1, 126.9, 125.9, 121.8, 115.8 (d, ² J _CF = 21.7Hz), 114.1, 107.8, 55.4, 44.4. HRMS-ESI: m/z calculated for C ₂ 5H ₂₂ N ₆ O ₃ F[M+H] ⁺ 473.1732,found473.1722.

Compound A12: 4-(2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)guanidino)piperidine-1-carboxylic acid ethyl ester hydrochloride is characterized as follows:

White solid; total yield 15%; Mp: 186-188°C; ¹ H NMR (600MHz, DMSO-d ₆ , ppm) δ: 11.16 (s, 1H), 9.62 (d, J = 7.6Hz, 1H), 8.98(s,2H),8.89(s,1H,aromatic),8.69(d,J＝7.8Hz,1H,aromatic),8.41(dd,J＝8.2,2.3Hz,1H,aromatic),8.37(s, 1H,aromatic),8.22(d,J＝8.6Hz,2H,aromatic),7.86(t,J＝8.0Hz,1H,aromatic),7.10(d,J＝8.4Hz,2H,aromatic),4.05(q ,J＝7.1Hz,2H,O-CH ₂ -),3.96(overlap,3H,piperidinyl),3.86(s,3H,OCH ₃ ),3.12-2.89(m,2H,piperidinyl),2.09-1.95(m,2H,piperidinyl),1.58-1.42(m, 2H, piperidinyl), 1.19 (t, J = 7.1Hz, 3H, CH ₃ ); ¹³ C NMR (150MHz, DMSO-d ₆ ,ppm)δ:165.4,162.5,157.4,154.6,153.0,148.4,137.1,133.8,130.7,129.4,127.3,126.0,121.7,114.4,108.0,60.8,55.6,48.3,41.8,30.8, 14.6.HRMS-ESI :m/z calculated for C ₂₆ H ₃₀ N ₇ O ₅ [M+H] ⁺ 520.2303,found 520.2313.

Compound A13: 4-((2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)guanidino)methyl)benzoic acid hydrochloride is characterized as follows:

White solid; total yield 17%; Mp: 232-233°C; ¹ H NMR (400 MHz, DMSO-d ₆ ,ppm)δ:11.47(s,1H),9.99(s,1H),9.11(s,2H),8.88(s,1H,aromatic),8.58(d,J＝7.9Hz,1H,aromatic),8.34(dd,J＝8.2,1.5Hz,1H,aromatic),8.29(s,1H,aromatic),8.00(d, J＝8.3Hz,2H,aromatic),7.92(d,J＝8.6Hz,2H,aromatic),7.76(t,J＝8.0Hz,1H,aromatic),7.65(d,J＝8.3Hz,2H,aromatic),6.84(d,J＝8.8Hz,2H,aromatic),4.79(d,J＝5.2Hz,2H,N-CH ₂ ), 3.79(s,3H,OCH ₃ ^; 9,125.9,121.8,114.2,107.8,55.4,44.9.HRMS-ESI:m/z calculated for _{C 26} H ₂₃ N ₆ O ₅ [M+H] ⁺ 499.1724, found 499.1716.

Compound A14: 1-(4-methoxybenzyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 23%; Mp: 250°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.41 (s, br, 1H), 9.89 (s, 1H), 9.09-8.75 (overlap) ,3H),8.50(d,J＝7.3Hz,1H,aromatic),8.34(d,J＝8.1Hz,1H,aromatic),8.23(s,1H,aromatic),7.89-7.76(m,2H,aromatic) ),7.72(t,J＝7.8Hz,1H,aromatic),7.48(d,J＝8.2Hz,2H,aromatic),7.00(d,J＝8.3Hz,2H,aromatic),6.81(d,J＝ 6.4Hz,2H,aromatic),4.57(s,2H,N-CH ₂ ),3.79(overlap,6H); ¹³ C NMR(100MHz,DMSO-d6,ppm)δ:164.50,162.37,162.18,159.27,157.31,153.52,148.22,136.82,133.50,130.39,130.21,129.07,12 7.34,126.74 ,125.90,121.74,114.38,114.10,107.47,55.37,55.24,45.02.HRMS-ESI:m/z calculated for C ₂₆ H ₂₅ N ₆ O ₄ [M+H] ⁺ 485.1932, found 485.1922.

Compound A15: 2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)-1-(4-(trifluoromethoxy)benzyl)guanidine hydrochloride is characterized as follows:

White solid; total yield 14%; Mp: 170°C; ¹ H NMR (600MHz, DMSO-d ₆ , ppm) δ: 11.41 (s, br, 1H), 9.94 (s, 1H), 9.08 (s, 2H) ),8.91(s,1H,aromatic),8.60(d,J＝7.6Hz,1H,aromatic),8.41-8.27(overlap,2H,aromatic),8.02(d,J＝7.6Hz,2H,aromatic), 7.78(t,J＝7.8Hz,1H,aromatic),7.67(d,J＝7.9Hz,2H,aromatic),7.45(d,J＝7.9Hz,2H,aromatic),6.94(d,J＝8.0Hz ,2H,aromatic),4.75(s,2H,N-CH ₂ ), 3.82 (s, 3H, OCH ₃ ); ¹³ C NMR (150MHz, DMSO-d ₆ , ppm) δ: 165.0, 162.5, 162.4, 157.4, 154.0, 148.3, 148.0, 137.0, 135.5, 133.7, 130.5, 130.1 ,129.3,127.0,125.9,121.9,121.4,120.9(q, ^α J _CF =259.4Hz),119.2,114.3,107.9,55.5,45.3.HRMS-ESI:m/z calculated for C ₂₆ H ₂₂ N ₆ O ₄ F ₃ [M+H] ⁺ 539.1649,found 539.1641.

Compound A16: (1r,4r)-4-(2-(4-(4-methoxyphenyl))-6-(3-nitrophenylpyrimidin-2-yl)guanidino)cyclohexane-1-carboxylic acid hydrochloride is characterized as follows:

White solid; total yield 18%; Mp: 311-313°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 12.28 (s, br, 1H), 11.10 (s, 1H), 9.60 (s ,1H),8.80(overlap,3H),8.66(d,J＝7.8Hz,1H,aromatic),8.37(d,J＝8.1Hz,1H,aromatic),8.30(s,1H,aromatic),8.15( d,J＝8.3Hz,2H,aromatic),7.83(t,J＝7.9Hz,1H,aromatic),7.06(d,J＝8.4Hz,2H,aromatic),3.85(s,3H,OCH ₃ ), 3.62(overlap,1H,substituted cyclohexyl),2.33-2.20(m,1H,substituted cyclohexyl),2.10-2.01(m,2H,substituted cyclohexyl),2.02-1.88(m,2H,substituted cyclohexyl),1.56-1.27(m,4H,substituted cyclohexyl) ; ¹³ C NMR (100MHz, DMSO-d ₆ ,ppm)δ:176.4,165.3,162.6,162.2,157.4,152.9,148.4,137.0,133.8,130.7,129.4,127.2,126.1,121.5,114.4,107.8,55.6,49.8,41.1,31.0 ,27.2.HRMS-ESI :m/z calculated for C ₂₅ H ₂₇ N ₆ O ₅ [M+H] ⁺ 491.2037,found 491.2027.

Compound A17: 2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)-1-(4-(trifluoromethyl)benzyl)guanidine hydrochloride is characterized as follows:

White solid; total yield 16%; Mp: 157-159°C; ¹ H NMR (400 MHz, DMSO-d ₆ , ppm) δ: 11.46 (s, 1H), 9.98 (s, 1H), 9.11 (s, 2H), 8.92 (s, 1H, aromatic), 8.62 (d, J = 7.8 Hz, 1H, aromatic), 8.41-8.29 (m, 2H, aromatic), 8.04 (d, J = 8.5 Hz, 2H, aromatic), 7.86-7.65 (m, 5H, aromatic), 6.93 (d, J = 8.4 Hz, 2H, aromatic), 4.83 (d, J = 5.6 Hz, 2H, N-CH ₂ ), 3.82 (s, 3H, OCH ₃ ); ¹³ C NMR (100 MHz, DMSO-d ₆ ,ppm)δ:165.0,162.6,162.4,157.4,154.1,148.3,140.8,137.0,133.7,130.5,129.3,128.8,128.7(q, ^β J _CF =32.7Hz),127.0,125.9,125.7(q, ^γ J _CF = 3.8Hz), 125.6 (q, ^α J _CF = 271.9Hz), 121.9, 114.2, 108.0, 55.4, 44.5. HRMS-ESI: m/zcalced for C ₂₆ H ₂₂ N ₆ O ₃ F ₃ [M+H] ⁺ 523.1700, found 523.1695.

Compound A18: 1-(3,4-dimethoxybenzyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)-pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 23%; Mp: 158-160°C; ¹ H NMR (600MHz, DMSO-d ₆ ,ppm)δ:11.36(s,1H),9.97(s,1H),9.04-8.66(overlap,3H),8.52(d,J＝7.5Hz,1H,aromatic),8.36(d,J＝8.1Hz,1H,aromatic),8.29(s,1H,aromatic),7.85(d,J＝7.7Hz,2H,ar omatic),7.74(t,J＝7.9Hz,1H,aromatic),7.21(s,1H,aromatic),7.08(d,J＝8.1Hz,1H,aromatic),7.01(d,J＝8.1Hz,1H,aromatic),6.83(d,J＝8.1Hz,2H,aromatic),4.55(s,2H,N-CH ₂ ),3.81(s,3H,OCH),3.78(s,3H,OCH),3.70(s,3H,OCH); ¹³ C NMR(150MHz,DMSO-d ₆ ,ppm)δ:164.7,162.6,162.3,157.4,153.5,149.0,148.8,148.3,137.0, 133.6,130.5,129.2,127.7,126.9,126.0,121.9,121.1,114.2,112.4,111.9,107.7,55.6,55.6,55.4,45.5.HRMS-ESI:m/z calculated for C ₂₇ H ₂₇ N ₆ O ₅ [M+ H] ⁺ 515.2037, found 515.2029.

Compound A19: 2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)-1-methylguanidine hydrochloride is characterized as follows:

White solid; total yield 23%; Mp: 110°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.23 (s, br, 1H), 9.29 (s, 1H), 9.02-8.76 (overlap) ,3H),8.63(d,J＝7.6Hz,1H,aromatic),8.35(d,J＝8.0Hz,1H,aromatic),8.26(s,1H,aromatic),8.14(d,J＝8.3Hz, 2H,aromatic),7.79(t,J＝7.9Hz,1H,aromatic),7.03(d,J＝8.5Hz,2H,aromatic),3.84(s,3H,OCH ₃ ),3.00(s,3H,CH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ ,ppm)δ:165.0,162.4,162.2,157.3,154.7,148.3,136.9,133.7,130.5,129.4,127.1,125.9,121.7,114.3,107.6,55.5,28.4.HRMS-ESI:m/z calculated for C _{1 9} H ₁₉ N ₆ O ₃ [M+H] ⁺ 379.1513,found379.1510.

Compound A20: 1-(2-(6-methoxy-1H-indol-3-yl))ethyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 15%; Mp: 187-188°C; ¹ H NMR (600 MHz, DMSO-d ₆ ,ppm)δ:11.30(br,1H),10.76(s,1H),9.70(s,1H),9.01-8.60(overlap,3H),8.54(d,J＝7.7Hz,1H,aromatic),8.35(dd,J＝8.2,2.3Hz,1H,aromatic),8.27(s,1H,aromatic),7.9 9(d,J＝8.3Hz,2H,aromatic),7.74(t,J＝8.0Hz,1H,aromatic),7.28-7.12(m,2H,aromatic),7.06(d,J＝2.4Hz,1H,aromatic),6.89(d,J＝8.3Hz,2H,aromatic),6.68(dd,J＝8.7,2.4Hz,1H, aromatic),3.80(s,3H,OCH ₃ ), 3.74 (s, 3H, OCH ₃ ), 3.37 (overlap, 2H, CH ₂ ), 3.07 (t, J = 6.9Hz, 2H, CH ₂ ); ¹³ C NMR (150MHz, DMSO-d ₆ ,ppm)δ:165.0,162.3,162.3,157.5,154.0,153.1,148.3,137.0,133.6,131.4,130.5,129.3,129.2,127.3,127.1,125.8,123.9,121.7,114.3,1 12.2,111.3,110.1,107.6,100.0,55.5,55.3,42.1,24.1.HRMS-ESI:m/zcalced for C ₂₉ H ₂₈ N ₇ O ₄ [M+H] ⁺ 538.2197, found 538.2194.

Compound A21: 2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)-1-(3,4,5-trimethoxybenzyl)guanidine hydrochloride is characterized as follows:

White solid; total yield 19%; Mp: 251-252°C; ¹ H NMR (600MHz, DMSO-d ₆ , ppm) δ: 11.40 (s, 1H), 10.02 (s, 1H), 8.99 (s, 2H) ),8.89(s,1H,aromatic),8.54(d,J＝7.5Hz,1H,aromatic),8.35(d,J＝8.0Hz,1H,aromatic),8.30(s,1H,aromatic),7.93( d,J＝8.0Hz,2H,aromatic),7.74(t,J＝7.9Hz,1H,aromatic),6.93(s,2H,aromatic),6.88(d,J＝8.3Hz,2H,aromatic),4.60 (d,J＝4.4Hz,2H,N-CH ₂ ), 3.81 (s, 3H, OCH ₃ ), 3.74 (s, 6H, 2OCH ₃ ), 3.68 (s, 3H, OCH ₃ ); ¹³ C NMR (150MHz, DMSO-d ₆ , ppm) δ: 164.8, 162.6 ,162.4,157.4,153.7,153.2,148.3,137.3,137.0,133.5,131.1,130.4,129.2,126.9,125.9,121.8,114.2,107.8,106.0,60.0,56.0,55.4,45. 7.HRMS-ESI:m/z calculated for C ₂₈ H ₂₉ N ₆ O ₆ [M+H] ⁺ 545.2143, found 545.2133.

Compound A22: 1-(4-bromobenzyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 15%; Mp: 177°C; ¹ H NMR (600 MHz, DMSO-d ₆ ,ppm)δ:11.40(s,1H),9.93(s,1H),9.00(s,2H),8.90(s,1H,aromatic),8.58(d,J＝7.7Hz,1H,aromatic),8.37(d,J＝8.1Hz,1H,aromatic),8.31(s,1H,aromatic),7.93(d,J＝8. 0Hz, 2H, aromatic), 7.78 (t, J = 8.0Hz, 1H, aromatic), 7.65 (d, J = 8.2Hz, 2H, aromatic), 7.52 (d, J = 8.2Hz, 2H, aromatic), 6.90 (d, J = 8.5Hz, 2H, aromatic), 4.67 (d, J = 5.1Hz, 2H, N-CH ₂ ), 3.84 (s,3H,OCH ₃ ); ¹³ C NMR (150MHz, DMSO-d ₆ , ppm) δ: 165.3, 163.0, 162.8, 157.8, 154.3, 148.8, 137.4, 135.6, 134.0, 132.3, 131.1, 130.9, 129.6, 127.3, 126.4, 122.3, 121.9,114.7,108.3,56.0,45.0.HRMS-ESI:m/z calculated for C ₂₅ H ₂₂ N ₆ O ₃ Br[M+H] ⁺ 533.0931,found533.0922.

Compound A23: 1-(4-chlorobenzyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 19%; Mp: 147°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.40 (s, 1H), 9.92 (s, 1H), 9.02 (s, 2H), 8.87(s,1H,aromatic),8.56(d,J＝7.7Hz,1H,aromatic),8.36(dd,J＝8.1,1.4Hz,1H,aromatic),8.28(s,1H,aromatic),7.89( d,J＝8.2Hz,2H,aromatic),7.77(t,J＝8.0Hz,1H,aromatic),7.62-7.48(overlap,4H),6.86(d,J＝8.5Hz,2H,aromatic),4.68 (d,J＝5.0Hz,2H,N-CH ₂ ), 3.82 (s, 3H, OCH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 164.8, 162.5, 162.3, 157.3, 153.8, 148.3, 136.9, 134.8, 133.6, 132.9, 130.4, 130.3 ,129.2,128.9,126.9,125.9,121.8,114.2,107.8,55.5,44.5.HRMS-ESI:m/z calculated for C ₂₅ H ₂₂ N ₆ O ₃ Cl[M+H] ⁺ 489.1436, found 489.1429.

Compound A24: 1-(4-hydroxy-3-methoxybenzyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 21%; Mp: 198-199°C; ¹ H NMR (600 MHz, DMSO-d ₆ ,ppm)δ:11.33(s,1H),9.95(s,1H),9.33(s,1H),8.93-8.70(overlap,3H),8.52(d,J＝7.6Hz,1H,aromatic),8.35(d,J＝8.2Hz,1H,aromatic),8.28(s,1H,aromatic),7.82(d,J ＝8.1Hz,2H,aromatic),7.74(t,J＝7.9Hz,1H,aromatic),7.18(s,1H,aromatic),6.96(d,J＝7.9Hz,1H,aromatic),6.90-6.73(m,3H),4.51(d,J＝4.1Hz,2H,N-CH ₂ ),3.82(s,3H,OCH ₃ ) ,3.73(s,3H,OCH ₃ ); ¹³ C NMR (150MHz, DMSO-d ₆ , ppm) δ: 164.6, 162.6, 162.3, 157.3, 153.4, 148.3, 147.9, 146.9, 136.9, 133.5, 130.4, 129.1, 126.8, 125.9, 121.8, 12 1.5,115.7,114.2,113.0,107.6,55.7,55.5,45.7.HRMS-ESI:m/z calculated for C ₂₆ H ₂₅ N ₆ O ₅ [M+H] ⁺ 501.1881, found 501.1873.

Compound A25: 1-(furan-2-ylmethyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 17%; Mp: 186°C; ¹ H NMR (400 MHz, DMSO-d ₆ ,ppm)δ:11.35(s,1H),9.96(s,1H),9.08(s,2H),8.87(s,1H,aromatic),8.56(d,J＝7.8Hz,1H,aromatic),8.35(dd,J＝8.2,1.5Hz,1H,aromatic),8.28(s,1H,aromatic),8.02(d, J＝8.6Hz,2H,aromatic),7.83-7.71(overlap,2H),6.97(d,J＝8.8Hz,2H,aromatic),6.65(d,J＝3.0Hz,1H,aromatic),6.59-6.48(m,1H,aromatic),4.74(d,J＝5.2Hz,2H,N-CH ₂ ),3.84(s,3 H,OCH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 164.8, 162.4, 157.2, 153.9, 148.7, 148.3, 143.8, 136.9, 133.5, 130.5, 129.2, 126.9, 125.9, 121.8, 114.3, 110.8, 109. 4,107.8,55.5,38.4.HRMS-ESI:m/z calculated for C ₂₃ H ₂₁ N ₆ O ₄ [M+H] ⁺ 445.1619, found 445.1614.

Compound A26: 2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)-1-(4-methyl-benzyl)guanidine hydrochloride is characterized as follows:

Pale yellow solid; total yield 19%; Mp: 190-191°C; ¹ H NMR (600 MHz, DMSO-d ₆ ,ppm)δ:11.37(s,1H),9.91(s,1H),8.95(s,2H),8.88(s,1H,aromatic),8.52(d,J＝7.6Hz,1H,aromatic),8.36(d,J＝8.1Hz,1H,aromatic),8.29(s,1H,aromatic),7.92(d,J＝8. 4Hz,2H,aromatic),7.74(t,J＝8.0Hz,1H,aromatic),7.41(d,J＝7.8Hz,2H,aromatic),7.25(d,J＝7.8Hz,2H,aromatic),6.86(d,J＝8.5Hz,2H,aromatic),4.61(s,2H,N-CH ₂ ),3.82(s,3H,OCH ₃ ), 2.34 (s, 3H, CH ₃ ); ¹³ C NMR (150MHz, DMSO-d ₆ , ppm) δ: 164.8, 162.5, 162.3, 157.3, 153.7, 148.3, 137.5, 136.9, 133.5, 132.5, 130.4, 129.5, 129.2, 1 28.4,126.9,125.9,121.8,114.2,107.7,55.5,45.1,20.8.HRMS-ESI:m/z calculated for C ₂₆ H ₂₅ N ₆ O ₃ [M+H] ⁺ 469.1983, found 469.1985.

Compound A27: 2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)-1-(4-phenoxybenzyl)guanidine hydrochloride is characterized as follows:

White solid; total yield 22%; Mp: 158-160°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.37 (s, 1H), 9.92 (s, 1H), 9.05-8.78 (overlap) ,3H),8.66-8.48(m,1H,aromatic),8.44-8.28(m,2H,aromatic),8.05-7.87(m,2H,aromatic),7.80-7.66(m,1H,aromatic),7.59- 7.48(m,2H,aromatic),7.45 -7.32(m,2H,aromatic),7.23-7.10(m,1H,aromatic),7.08-6.90(m,6H,aromatic),4.63(d,J＝4.9Hz ,2H,N-CH ₂ ), 3.83 (s, 3H, OCH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 164.9, 162.6, 162.4, 157.3, 156.7, 156.3, 153.7, 151.0, 148.3, 137.0, 133.6, 130.5,130.2,130.1,129.2,127.0,125.9,123.8,121.9,118.8,118.8,114.3,107.9,55.5,44.6.HRMS-ESI:m/z calculated for C ₃₁ H ₂₇ N ₆ O ₄ [M+H] ⁺ 547.2088,found547.2080.

Compound A28: 1-(2-fluorobenzyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 20%; Mp: 144-146°C; ¹ H NMR (400 MHz, DMSO-d ₆ ,ppm)δ:11.41(s,1H),9.94(s,1H),9.20-8.89(overlap,3H),8.63(d,J＝7.8Hz,1H,aromatic),8.47-8.35(m,2H,aromatic),8.10(d,J＝8.5Hz,2H,aromatic),7.82(t,J＝8.0Hz, 1H,aromatic),7.65(t,J＝7.4Hz,1H,aromatic),7.53-7.40(m,1H,aromatic),7.39-7.21(m,2H,aromatic),7.00(d,J＝8.7Hz,2H,aromatic),4.77(d,J＝5.4Hz,2H,N-CH ₂ ),3.85(s,3H,OCH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 165.1, 162.7, 162.4, 161.6 (d, ¹ J _CF = 241.0Hz), 157.4, 154.0, 148.4, 137.1, 133.7, 130.6 (d, ^{3 J CF = 7.7Hz), 130.6, 130. 2 (d, 3} _{J CF} ⁼ _4.4Hz ), 129.3, 127.1, 126.0, 124.9 (d, ⁴ J _CF = 3.7Hz), 122.9 (C, d, ² J _CF = 14.9Hz), 121.9, 115.8 (CH, d, ² J _CF ＝20.8Hz),114.4,108.1,55.5,39.9(CH ₂ ,d, ³ J _CF ＝3.9Hz).HRMS-ESI:m/zcalced forC ₂₅ H ₂₂ FN ₆ O ₃ [M+H] ⁺ 473.1732, found 473.1731.

Compound A29: 4-(2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)guanidino)butyric acid hydrochloride was characterized as follows:

White solid; total yield 20%; Mp: 77°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.24 (s, 1H), 9.58 (s, 1H), 9.02-8.80 (overlap, 3H) ),8.63(d,J＝7.8Hz,1H,aromatic),8.35(dd,J＝8.2,2.3Hz,1H,aromatic),8.27(s,1H,aromatic),8.14(d,J＝8.6Hz, 2H,aromatic),7.80(t,J＝8.0Hz,1H,aromatic),7.03(d,J＝8.7Hz,2H,aromatic),3.84(s,3H,OCH ₃ ),3.40(overlap,2H,carboxypropyl ,CH ₂ ), 2.40 (t, J＝7.3Hz, 2H, carboxypropyl, CH ₂ ), 1.89 (p, J＝7.2Hz, 2H, carboxypropyl, CH ₂ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ :174.0,165.1,162.5,162.3,157.3,154.0,148.3,137.0,133.7,130.5,129.3,127.1,125.9,121.7,114.3,107.7,55.5,40.8,30.9,23.7.HRMS-ESI :m/z calculated for C ₂₂ H ₂₃ N ₆ O ₅ [M+H] ⁺ 451.1724,found 451.1717.

Compound A30: N-(2-(2-(4-(4-methoxyphenyl))-6-(3-nitrophenylpyrimidin-2-yl)guanidino)ethyl)acetamide hydrochloride is characterized as follows:

White solid; total yield 15%; Mp: 207-208°C; ¹ H NMR (400 MHz, DMSO-d ₆ , ppm) δ: 11.23 (s, 1H), 9.55 (d, J = 7.0 Hz, 1H), 9.02-8.85 (overlap, 3H), 8.67 (d, J = 7.9 Hz, 1H, aromatic), 8.42-8.28 (m, 3H, aromatic), 8.18 (d, J = 8.6 Hz, 2H, aromatic), 7.83 (t, J = 8.0 Hz, 1H, aromatic), 7.10-6.99 (m, 2H, aromatic), 3.85 (s, 3H, OCH ₃ ), 3.58-3.46 (m, 2H, CH ₂ ), 3.38-3.30 (m, 2H, CH ₂ ), 1.76 (s, 3H, CO-CH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 170.2, 165.1, 162.5, 162.4, 157.3, 154.3, 148.3, 137.0, 133.7, 130.6, 129.4, 127.1, 125.9 ,121.8,114.4,107.8,55.5,41.2,37.5,22.5.HRMS-ESI:m/zcalced for C ₂₂ H ₂₄ N ₇ O ₄ [M+H] ⁺ 450.1884, found 450.1876.

Compound A31: 1-(3-hydroxypropyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 18%; Mp: 129-130°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.15 (s, 1H), 9.47 (s, 1H), 8.92-8.68 (overlap) ,3H),8.58(d,J＝7.9Hz,1H,aromatic),8.31(dd,J＝8.1,2.2Hz,1H,aromatic),8.18(s,1H,aromatic),8.08(d,J＝8.4 Hz,2H,aromatic),7.76(t,J＝8.0Hz,1H,aromatic),6.98(d,J＝8.5Hz,2H,aromatic),4.84(s,1H,OH),3.82(s,3H, OCH ₃ ),3.56(t,J＝6.0Hz,2H,hydroxypropyl,CH ₂ ),3.45-3.38(overlap,2H,hydroxypropyl,CH ₂ ),1.81(p,J＝6.4Hz,2H,hydroxypropyl,CH ₂ ); ^13C NMR (100MHz, DMSO-d ₆ , ppm) δ: 13C NMR (101MHz, DMSO) δ164.9,162.4,162.1,157.2,153.9,148.2,136.8,133.6,130.4,129.3,127.0,125.8,121.6,114.2 , 107.5,57.9,55.5,39.3,31.0.HRMS-ESI:m/z calculated for C ₂₁ H ₂₃ N ₆ O ₄ [M+H] ⁺ 423.1775, found 423.1771.

Compound A32: 1-(2-hydroxyethyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 17%; Mp: 136-138°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.15 (br, 1H), 9.78 (s, 1H), 8.87 (s, 1H) ,aromatic),8.79-8.53(overlap,3H),8.34(dd,J＝8.2,2.4Hz,1H,aromatic),8.26(s,1H,aromatic),8.24-8.17(m,2H,aromatic),7.78 (t,J＝8.0Hz,1H,aromatic),7.02(d,J＝8.9Hz,2H,aromatic),5.39(br,1H,OH),3.83(s,3H,OCH ₃ ), 3.70 (t, J = 5.2Hz, 2H, hydroxyethyl), 3.50 (t, J = 5.2Hz, 2H, hydroxyethyl); ¹³ CNMR (100MHz, DMSO-d ₆ , ppm) δ: 165.0, 162.4, 162.3, 157.3,154.1,148.3,136.9,133.7,130.5,129.5,126.9,125.9,121.8,114.3,107.5,58.6,55.5,44.2.HRMS-ESI:m/z calculated for C ₂₀ H ₂₁ N ₆ O ₄ [M+H ] ⁺ 409.1619, found 409.1616.

Compound A33: 1-benzyl-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 16%; Mp: 175°C; ¹ H NMR (400 MHz, DMSO-d ₆ ,ppm)δ:11.44(s,1H),9.98(s,1H),9.09(s,2H),8.87(t,J＝2.0Hz,1H,aromatic),8.53(d,J＝7.8Hz,1H,aromatic),8.34(dd,J＝8.2,2.3Hz,1H,aromatic),8.29(s,1H,aromatic), 7.95(d,J＝8.6Hz,2H,aromatic),7.74(t,J＝8.0Hz,1H,aromatic),7.53(dt,J＝5.9,1.6Hz,2H, aromatic),7.49-7.37(m,3H,aromatic),6.88(d,J＝8.7Hz,2H,aromatic),4.69(d,J＝4.2Hz,2H ,N-CH ₂ ), 3.81 (s, 3H, OCH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 164.8, 162.4, 162.3, 157.3, 153.9, 148.3, 136.9, 135.7, 133.5, 130.5, 129.2, 129.0, 128.3, 128.2,126.9,125.9,121.8,114.2,107.7,55.5,45.2.HRMS-ESI:m/z calculated for C ₂₅ H ₂₃ N ₆ O ₃ [M+H] ⁺ 455.1826, found 455.1818.

Compound A34: 1-ethyl-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 24%; Mp: 239-241°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.18 (s, 1H), 9.50 (t, J = 5.3Hz, 1H), 8.92-8.73(overlap,3H),8.63(d,J＝7.9Hz,1H,aromatic),8.35(d,J＝8.2Hz,1H,aromatic),8.25(s,1H,aromatic),8.12(d, J＝8.4Hz,2H,aromatic),7.79(t,J＝8.0Hz,1H,aromatic),7.02(d,J＝8.4Hz,2H,aromatic),3.84(s,3H,OCH ₃ ), 3.43-3.33 (overlap, 2H), 1.28 (t, J＝7.1Hz, 3H, C-CH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 165.0, 162.4, 162.1, 157.3 ,153.7,148.2,136.9,133.7,130.5,129.3,127.0,125.9,121.6,114.3,107.6,55.5,36.5,13.7.HRMS-ESI:m/z calculated for C ₂₀ H ₂₁ N ₆ O ₃ [M+H ] ⁺ 393.1670, found 393.1667.

Compound A35: 1-methoxy-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 20%; Mp: 236-238°C; ¹ H NMR (400MHz, Methanol-d ₄ , ppm) δ: 8.78 (t, J=1.8Hz, 1H, aromatic), 8.47 (d, J ＝7.6Hz,1H,aromatic),8.31(dd,J＝8.1,2.2Hz,1H,aromatic),8.08-7.90(m,3H,aromatic),7.72(t,J＝7.9Hz,1H,aromatic), 6.97 (d, J=8.4Hz, 2H, aromatic), 3.94 (s, 3H, OCH ₃ ), 3.85 (s, 3H, OCH ₃ ); ¹³ C NMR (100MHz, Methanol-d ₄ ,ppm)δ:167.4,164.5,164.3,158.6,156.8,150.0,138.5,134.3,131.6,130.4,128.4,126.9,122.9,115.5,109.0,65.7,56.1.HRMS-ESI:m/zcalced for C ₁₉ H ₁₉ N ₆ O ₄ [M+H] ⁺ 395.1462, found 395.1457.

Compound A36: 1-(4-bromophenyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 19%; Mp: 266-268°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.60 (s, 1H), 11.44 (s, 1H), 9.46 (s, 1H) ),9.12-8.82(overlap,2H),8.71(d,J＝7.8Hz,1H,aromatic),8.47-8.32(overlap,2H,aromatic),8.23(d,J＝8.5Hz,2H,aromatic), 7.85(t,J＝8.0Hz,1H,aromatic),7.73(d,J＝8.4Hz,2H,aromatic),7.42(d,J＝8.4Hz,2H,aromatic),7.08(d,J＝8.5Hz ,2H,aromatic),3.85(s,3H,OCH _{3 ;} ^120.3 ,114.4,108.3,55.6.HRMS-ESI:m/z calculated forC ₂₄ H ₂₀ N ₆ O ₃ Br[M+H] ⁺ 519.0775, found 519.0767.

Compound A37: 1-(2-cyanoethyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 14%; Mp: 115-118°C; ¹ H NMR (600MHz, DMSO-d ₆ , ppm) δ: 11.43 (br, 1H), 9.81 (s, 1H), 9.03-8.72 (overlap) ,3H),8.62(d,J＝7.7Hz,1H,aromatic),8.36(d,J＝7.8Hz,1H,aromatic),8.27(s,1H,aromatic),8.18(d,J＝8.6Hz, 2H,aromatic),7.79(t,J＝7.9Hz,1H,aromatic),7.04(s,2H,aromatic),3.85(s,3H,OCH ₃ ),3.60(t,J＝7.1Hz,2H,CH ₂ ),2.69(t,J＝5.7Hz,2H,CH _{2 ;} ^107.6 ,55.5,37.6,33.2.HRMS-ESI:m/z calculated for C ₂₁ H ₂₀ N ₇ O ₃ [M+H] ⁺ 418.1622, found 418.1614.

Compound A38: 2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)-1-propylguanidine hydrochloride is characterized as follows:

White solid; total yield 25%; Mp: 115-118°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.08 (s, 1H), 9.43 (s, 1H), 8.84-8.69 (overlap) ,3H),8.61(d,J＝7.9Hz,1H,aromatic),8.34(dd,J＝8.1,1.6Hz,1H,aromatic),8.23(s,1H,aromatic),8.11(d,J＝8.8 Hz,2H,aromatic),7.79(t,J＝8.0Hz,1H,aromatic),7.02(d,J＝8.9Hz,2H,aromatic),3.83(s,3H,OCH ₃ ),3.30(q,J ＝6.7Hz,2H,CH ₂ ), 1.65 (h, J＝7.3Hz, 2H, CH ₂ ), 0.96 (t, J＝7.4Hz, 3H, CH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) 165.2, 162.6, 162.3 ,157.4,153.9,148.3,137.0,133.8,130.7,129.4,127.1,126.0,121.6,114.4,107.7,55.6,43.2,21.6,11.3.HRMS-ESI:m/z calcedfor C ₂₁ H ₂₃ N ₆ O ₃ [ M+H] ⁺ 407.1826, found 407.1823.

Compound A39: 1-cyclohexyl-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 21%; Mp: 229-230°C; ¹ H NMR (600 MHz, DMSO-d ₆ , ppm) δ: 11.21 (s, 1H), 9.65 (d, J = 7.1 Hz, 1H), 8.93-8.82 (overlap, 3H), 8.69 (d, J = 7.7 Hz, 1H, aromatic), 8.40 (d, J = 8.2 Hz, 1H, aromatic), 8.35 (s, 1H, aromatic), 8.20 (d, J = 8.3 Hz, 2H, aromatic), 7.85 (t, J = 7.9 Hz, 1H, aromatic), 7.09 (d, J = 8.3 Hz, 2H, aromatic), 3.86 (s, 3H, OCH ₃ ),3.76-3.62(m,1H,cyclohexyl),2.07-1.91(m,2H,cyclohexyl),1.82-1.69(m,2H,cyclohexyl),1.64-1.52(m,1H,cyclohexyl),1.43-1.32(m,4H,cyclohexyl),1.28-1.20(m,1H, cyclohexyl); ¹³ C NMR (150MHz, DMSO-d ₆ ,ppm)δ:165.7,163.0,162.7,157.9,153.3,148.8,137.6,134.3,131.1,129.8,127.7,126.4,122.0,114.8,108.3,56.0,50.4,32.4,25.1,24.4. HRMS-ESI: m/z calculated for C ₂₄ H ₂₇ N ₆ O ₃ [M+H] ⁺ 447.2139, found 447.2132.

Compound A40: 1-(3-bromopropyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 18%; Mp: 279-280°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 9.37 (s, 1H), 9.09 (s, 1H), 8.98 (s, 1H) ),8.72(d,J＝7.9Hz,1H,aromatic),8.49(s,1H,aromatic),8.44-8.35(m,1H,aromatic),8.29(d,J＝8.8Hz,2H,aromatic), 7.85(t,J＝8.0Hz,1H,aromatic),7.11(d,J＝8.8Hz,2H,aromatic),4.27-4.15(m,2H,CH ₂ ),3.86(s,3H,OCH ₃ ), 3.41(t,J＝5.7Hz,2H,CH ₂ ), 2.20-2.04 (m, 2H, CH ₂ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 165.2, 162.5, 162.4, 158.7, 154.2, 148.4, 137.2, 133.9, 130.6, 129.6, 127.3 ,126.0,122.0,114.4,108.4,55.6,44.6,38.4,20.5.HRMS-ESI:m/z calculated for C ₂₁ H ₂₂ BrN ₆ O ₃ [M+H] ⁺ 485.0931, found 485.0928.

Compound A41: 1-(3-chloropropyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 17%; Mp: 262-263°C; ¹ H NMR (600MHz, DMSO-d ₆ , ppm) δ: 9.27 (s, 1H), 9.17-8.80 (overlap, 3H), 8.74 (d ,J＝7.8Hz,1H,aromatic),8.53(s,1H,aromatic),8.45-8.39(m,1H,aromatic),8.32(d,J＝8.8Hz,2H,aromatic),7.88(t,J ＝8.0Hz,1H,aromatic),7.13(d,J＝8.8Hz,2H,aromatic),4.24(t,J＝5.8Hz,2H,CH ₂ ),3.87(s,3H,OCH ₃ ),3.42( td,J＝6.0,2.6Hz,2H,CH ₂ ), 2.13 (p, J=5.8Hz, 2H, CH ₂ ); ¹³ C NMR (150MHz, DMSO-d ₆ , ppm) δ: 165.3, 162.6, 162.5, 158.8, 154.2, 148.5, 137.3, 133.9, 130.7, 129.7,127.4,126.0,122.1,114.5,108.5,55.6,44.6,38.4,20.5.HRMS-ESI:m/z calculated forC ₂₁ H ₂₂ ClN ₆ O ₃ [M+H] ⁺ 441.1436,found 441.1430.

Compound A42: 1-(4-hydroxybutyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 19%; Mp: 146-147°C; ¹ H NMR (400 MHz, DMSO-d ₆ , ppm) δ: 11.16 (s, 1H), 9.49 (d, J = 5.6 Hz, 1H), 8.95 -8.72 (overlap, 3H), 8.61 (d, J = 7.8 Hz, 1H, aromatic), 8.34 (dd, J = 8.2, 2.3 Hz, 1H, aromatic), 8.24 (s, 1H, aromatic), 8.11 (d, J = 8.5 Hz, 2H, aromatic), 7.79 (t, J = 8.0 Hz, 1H, aromatic), 7.01 (d, J = 8.7 Hz, 2H, aromatic), 3.83 (s, 3H, OCH ₃ ),3.49-3.33(overlap,4H,hydroxybutyl),1.68(p,J＝7.1Hz,2H,hydroxybutyl),1.62-1.46(m,2H,hydroxybutyl); ¹³ C NMR(100MHz,DMSO-d ₆ ,ppm)δ:165.0,162.4,162.2,157.3,153.8,148 .2,136.9,133.7,130.5,129.3,127.0,125.9,121.6,114.3,107.6,60.1,55.5,41.4,29.5,24.9.HRMS-ESI:m/z calculated for C ₂₁ H ₂₅ N ₆ O ₄ [M+H] ⁺ 437.19 32,found 437.1928.

Compound A43: 1-cyclopentyl-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 23%; Mp: 234°C; ¹ H NMR (600MHz, DMSO-d ₆ , ppm) δ: 11.17 (s, 1H), 9.73 (d, J = 7.4Hz, 1H), 8.91 ( s,2H),8.83(s,1H,aromatic),8.64(d,J＝7.8Hz,1H,aromatic),8.36(dd,J＝8.1,2.3Hz,1H,aromatic),8.29(s,1H, aromatic),8.15(d,J＝8.4Hz,2H,aromatic),7.82(t,J＝8.0Hz,1H,aromatic),7.05(d,J＝8.5Hz,2H,aromatic),4.11(q,J ＝6.3Hz,1H,cyclopentyl),3.85(s,3H,OCH ₃ ), 2.06 (dt, J=12.0, 6.3Hz, 2H, cyclopentyl), 1.80-1.66 (m, 2H, cyclopentyl), 1.64-1.50 (m, 4H, cyclopentyl); ¹³ C NMR (150MHz, DMSO-d ₆ ,ppm)δ:165.1,162.4,162.2,157.3,153.3,148.2,137.0,133.7,130.5,129.3,127.1,125.9,121.5,114.3,107.8,55.5,52.8,32.3,23.2.HRMS-ESI :m/zcalcedfor C ₂₃ H ₂₅ N ₆ O ₃ [M+H] ⁺ 433.1983,found 433.1976.

Compound A44: (S)-2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)-1-(tetrahydrofuran-3-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 16%; Mp: 188°C; ¹ H NMR (400 MHz, DMSO-d ₆ ,ppm)δ:11.20(s,1H),9.99(s,1H),9.15-8.76(overlap,3H),8.62(d,J＝7.9Hz,1H,aromatic),8.36(dd,J＝8.1,2.3Hz,1H,aromatic),8.28(s,1H,aromatic),8.15(d,J＝8.5Hz, 2H,aromatic),7.80(t,J＝8.0Hz,1H,aromatic),7.03(d,J＝8.7Hz,2H,aromatic),4.46(s,1H,tetrahydrofuranyl),3.97-3.67(overlap,7H),2.42-2.26(m,1H,tetrahydrofuranyl),1.96(dt,J＝1 2.8,6.5Hz,1H,tetrahydrofuranyl); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 165.0, 162.5, 162.3, 157.3, 153.5, 148.3, 137.0, 133.7, 130.5, 129.3, 127.0, 126.0, 121.7, 114.3, 107.7, 72.3, 6 6.2,55.6,52.3,32.5.HRMS-ESI:m/z calculated for C ₂₂ H ₂₃ N ₆ O ₄ [M+H] ⁺ 435.1775,found 435.1769.

Compound A45: 1-(4-chlorophenyl)-2-(4-(4-methoxyphenyl)-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; yield 22%; Mp: 219-221°C; ¹ H NMR (600 MHz, DMSO-d ₆ ,ppm)δ:11.60(s,1H),11.46(s,1H),9.47(s,1H),9.04-8.85(overlap,2H),8.70(d,J＝7.9Hz,1H,aromatic),8.41(s,1H,aromatic),8.38(dd,J＝8.1,2.3Hz,1H,aromatic),8.2 1(d,J＝8.6Hz,2H,aromatic),7.83(t,J＝8.0Hz,1H,aromatic),7.66-7.57(m,2H,aromatic),7.52-7.43(m,2H,aromatic),7.06(d,J＝8.7Hz,2H,aromatic),3.84(s,3H,OCH ₃ ); ¹³ C NMR (150 MHz,DMSO-d ₆ ,ppm)δ:165.3,162.6,162.4,157.2,153.6,148.3,137.0,133.8,133.0,131.9,130.6,130.0,129.5,127.1,127.1,126.0,121.8,114.4,108.2,5 5.6.HRMS-ESI:m/zcalced for C ₂₄ H ₂₀ N ₆ O ₃ Cl[M+H] ⁺ 475.1280,found 475.1274.

Compound A46: 1-(4-hydroxyphenyl)-2-(4-(3-nitrophenyl)-6-phenylpyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 25%; Mp: 341-342°C; ¹ H NMR (400 MHz, DMSO-d ₆ ,ppm)δ:11.53(s,1H),9.99-9.76(overlap,2H),9.06(s,2H),8.94(s,1H,aromatic),8.54(d,J＝7.8Hz,1H,aromatic),8.46-8.28(overlap,2H,aromatic),7.97(d,J＝7.7Hz,2H, _aromatic ) ¹³ C NMR(100MHz,DMSO-d ₆ , ppm)δ:165.2,163.1,157.7,157.5,153.5,148.4,136.9,134.7,133.6,132.0,130.6,129.9,129.0,127.4,126.1,125.5,122.0,115.8,108. 7,45.2.HRMS-ESI:m/z calculated for C ₂₄ H ₂₁ N ₆ O ₃ [M+H] ⁺ 441.1670, found 441.1673.

Compound A47: 1-(2-chlorobenzyl)-2-(4-(3-nitrophenyl)-6-phenylpyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 21%; Mp: 254-256°C; ¹ H NMR (400 MHz, DMSO-d ₆ ,ppm)δ:11.59(s,1H),9.96(s,1H),9.15(s,2H),8.97(s,1H,aromatic),8.63(d,J＝7.9Hz,1H,aromatic),8.47(s,1H,aromatic),8.38(dd,J＝8.2,2.3Hz,1H,aromatic),8.08(d _, ¹³ C NMR 0MHz, DMSO-d ₆ ,ppm)δ:165.4,163.1,157.5,154.1,148.4,136.9,134.9,133.7,133.2,132.9,132.0,130.6,130.2,130.2,129.8,129.0,127.8,127.5,126.1,1 22.0,109.0,43.3.HRMS-ESI:m/z calculated for C ₂₄ H ₂₀ N ₆ O ₂ Cl[M+H] ⁺ 459.1331,found459.1323.

Compound A48: 1-(4-bromobenzyl)-2-(4-(4-bromophenyl)-6-(4-methoxyphenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; yield 17%; Mp: 184-186°C; ¹ H NMR (400 MHz, DMSO-d ₆ , ppm) δ: 11.37 (br, 1H), 10.02 (s, 1H), 8.93 (s, 2H), 8.22 (s, 1H, aromatic), 8.02 (dd, J＝8.7, 2.8 Hz, 4H, aromatic), 7.68 (d, J＝8.2 Hz, 4H, aromatic), 7.51 (d, J＝8.0 Hz, 2H, aromatic), 6.98 (d, J＝8.4 Hz, 2H, aromatic), 4.64 (s, 2H, CH ₂ ), 3.86 (s, 3H, OCH ₃ ); ¹³ C NMR (100 MHz, DMSO-d ₆ ,ppm)δ:165.2,164.0,162.8,157.9,154.4,135.6,135.0,132.4,131.2,129.7,129.6,127.7,125.9,122.0,114.8,107.8,56.0,45.0.HRMS-ESI:m/z calculated for C ₂₅ H ₂₂ N ₅ OBr ⁸¹ Br[M+H] ⁺ 568.0165, found 568.0156.

Compound A49: 2-(4-(4-bromophenyl)-6-(4-methoxyphenyl)pyrimidin-2-yl)-1-(4-hydroxy-benzyl)guanidine hydrochloride is characterized as follows:

White solid; total yield 17%; Mp: 192-194°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.31 (s, 1H), 9.97 (s, 1H), 9.81 (s, 1H) ),8.81(s,2H),8.16(s,1H,aromatic),7.97(d,J＝8.2Hz,2H,aromatic),7.90(d,J＝8.5Hz,2H,aromatic),7.64(d, J＝8.2Hz,2H,aromatic),7.36(d,J＝8.0Hz,2H,aromatic),6.98-6.81(m,4H,aromatic),4.48(d,J＝4.6Hz,2H,CH ₂ ), 3.83(s,3H,OCH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ ,ppm)δ:164.5,163.5,162.2,157.7,157.3,153.4,134.3,131.8,130.3,129.2,129.0,127.0,125.5,125.3,115.8,114.3,107.0, 55.5,45.3 .HRMS-ESI:m/z calculated for C ₂₅ H ₂₃ N ₅ O ₂ Br[M+H] ⁺ 504.1030, found 504.1025.

Compound A50: 2-(4,6-bis(4-methoxyphenyl))pyrimidin-2-yl)-1-(4-bromobenzyl)guanidine hydrochloride is characterized as follows:

White solid; total yield 18%; Mp: 251-253°C; ¹ H NMR (600 MHz, DMSO-d ₆ , ppm) δ: 11.33 (s, 1H), 10.16 (s, 1H), 9.01 (s, 2H), 8.14-7.85 (overlap, 5H, aromatic), 7.67 (d, J＝7.6 Hz, 2H, aromatic), 7.52 (d, J＝7.9 Hz, 2H, aromatic), 7.09-6.76 (overlap, aromatic), 4.65 (s, 2H, CH ₂ ), 3.83, 3.84 (s, 6H, 2OCH ₃ ); ¹³ C NMR (150 MHz, DMSO-d ₆ ,ppm)δ:164.1,162.1,157.2,154.0,135.1,131.9,130.9,129.0,129.0,127.4,121.6,114.2,106.3,55.5,44.7.HRMS-ESI:m/z calculated for C ₂₆ H ₂₅ N ₅ O ₂ Br[M+H] ⁺ 518.1186,found 518.1176.

Compound A51: 2-(4,6-bis(4-bromophenyl))pyrimidin-2-yl)-1-(4-hydroxybenzyl)guanidine hydrochloride is characterized as follows:

White solid; total yield 14%; Mp: 268-270°C; ¹ H NMR (600 MHz, DMSO-d ₆ , ppm) δ: 11.43 (s, 1H), 9.84 (s, 1H), 9.77 (s, 1H), 8.87 (s, 2H), 8.27 (s, 1H, aromatic), 7.95 (d, J = 8.2 Hz, 4H, aromatic), 7.63 (d, J = 8.1 Hz, 4H, aromatic), 7.35 (d, J = 8.0 Hz, 2H, aromatic), 6.87 (d, J = 8.0 Hz, 2H, aromatic), 4.50 (d, J = 4.9 Hz, 2H, CH ₂ ); ¹³ C NMR (150 MHz, DMSO-d ₆ ,ppm)δ:164.0,157.7,157.4,153.4,134.1,131.9,130.1,129.2,125.8,125.4,115.7,108.0,45.2.HRMS-ESI:m/z calculated for C ₂₄ H ₂₀ N ₅ OBr ₂ [M+H] ⁺ 552. 0029, found 552.0020.

Compound A52: 2-(4,6-bis(4-bromophenyl))pyrimidin-2-yl)-1-(4-bromobenzyl)guanidine hydrochloride is characterized as follows:

White solid; total yield 17%; Mp: 317-319°C; ¹ H NMR (400 MHz, DMSO-d ₆ , ppm) δ: 11.53 (s, 1H), 9.93 (s, 1H), 9.03 (s, 2H), 8.23 (s, 1H, aromatic), 7.93 (d, J = 8.2 Hz, 4H, aromatic), 7.70-7.56 (overlap, 6H, aromatic), 7.53 (d, J = 8.0 Hz, 2H, aromatic), 4.66 (d, J = 5.2 Hz, 2H, CH ₂ ); ¹³ C NMR (100 MHz, DMSO-d ₆ ,ppm)δ:164.0,157.4,153.8,135.1,134.0,131.9,130.8,129.2,125.8,121.6,108.1,44.6.HRMS-ESI:m/z calculated forC ₂₄ H ₁₉ N ₅ OBr ₂ ⁸¹ Br[M+H] ⁺ 615.9 165,found 615.9162.

Compound A53: 2-(4,6-diphenylpyrimidin-2-yl)-1-(4-hydroxybenzyl)guanidine hydrochloride is characterized as follows:

White solid; total yield 20%; Mp: 277-280°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.37 (s, 1H), 10.00 (s, 1H), 9.80 (s, 1H) ,OH),8.85(s,2H),8.29(s,1H,aromatic),8.06(d,J＝7.6Hz,4H,aromatic),7.57(t,J＝7.3Hz,2H,aromatic),7.47( t,J＝7.6Hz,4H,aromatic),7.34(d,J＝8.4Hz,2H,aromatic),6.86(d,J＝8.4Hz,2H,aromatic),4.50(d,J＝4.2Hz,2H ,N-CH ₂ ); ¹³ C NMR (100MHz, DMSO-d ₆ ,ppm)δ:165.1,157.7,157.6,153.5,135.1,131.9,130.1,129.1,127.4,125.5,115.8,108.3,45.3.HRMS-ESI:m/z calculated for C ₂₄ H ₂₂ N ₅ O[M+ H] ⁺ 396.1819, found 396.1813.

Compound A54: 1-(4-hydroxyphenyl)-2-(4-(3-nitrophenyl)-6-(3,4,5-trimethoxyphenyl)-pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 23%; Mp: 226-228°C; ¹ H NMR (600MHz, DMSO-d ₆ , ppm) δ: 11.37 (s, br, 1H), 9.69-9.53 (overlap, 2H), 9.21 -8.82(overlap,3H),8.57(d,J＝7.5Hz,1H,aromatic),8.43-8.34(m,2H,aromatic),7.80(t,J＝8.0Hz,1H),7.46(s,2H ,aromatic),7.26(d,J＝8.5Hz,2H,aromatic),6.79(d,J＝8.5Hz,2H,aromatic),4.56(s,2H,N-CH ₂ ),3.84(s,6H, 2OCH ₃ ),3.75(s,3H,OCH ₃ ; ^108.8 _, 105.2,60.2,56.2,44.5.HRMS-ESI:m/z calculated for C ₂₇ H ₂₇ N ₆ O ₆ [M+H] ⁺ 531.1987, found 531.1984.

Compound A55: 1-(2-chlorobenzyl)-2-(4-(3-nitrophenyl)-6-(3,4,5-trimethoxyphenyl)-pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 21%; Mp: 213-215°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.54 (s, 1H), 9.81 (s, 1H), 9.23 (s, 2H) ),8.94(s,1H,aromatic),8.68(d,J＝7.7Hz,1H,aromatic),8.54-8.32(m,2H,aromatic),7.84(t,J＝8.0Hz,1H,aromatic), 7.67-7.57(m,1H,aromatic),7.56-7.45(m,3H,aromatic),7.43-7.31(m,2H,aromatic),4.81(d,J＝5.6Hz,2H,N-CH ₂ ), 3.84(s,6H,2OCH ₃ ), 3.75 (s, 3H, OCH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 165.3, 163.0, 157.3, 154.3, 153.2, 148.3, 140.1, 137.1, 133.9, 133.5, 132.4, 130.6 ,130.3,129.7,129.6,128.8,127.6,126.0,122.0,109.0,105.3,60.2,56.2,42.9.HRMS-ESI:m/z calculated for C ₂₇ H ₂₆ N ₆ O ₅ Cl[M+H] ⁺ 549.1648 , found 549.1639.

Compound A56: 2-(4-(3-nitrophenyl))-6-(3,4,5-trimethoxyphenyl)pyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 19%; Mp: 178°C; ¹ H NMR (400 MHz, DMSO-d ₆ , ppm) δ: 11.23 (s, 1H), 9.50 (d, J = 8.0 Hz, 1H), 9.09 (s, 2H), 8.88 (t, J = 2.0 Hz, 1H, aromatic), 8.74 (d, J = 7.9 Hz, 1H, aromatic), 8.48-8.35 (overlap, 2H, aromatic), 7.88 (t, J = 8.0 Hz, 1H, aromatic), 7.49 (s, 2H, aromatic), 3.91 (overlap, 9H), 3.77 (s, 3H, OCH ₃ ), 3.44 (td, J=11.5, 2.1Hz, 2H, tetrahydropyranyl), 2.08-1.94 (m, 2H, tetrahydropyranyl), 1.60-1.50 (m, 2H, tetrahydropyranyl); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 165.4, 162.8, 157. 3,153.3,152.8,148.3,141.0,137.1,134.0,130.6,130.4,126.0,121.7,108.9,105.3,65.4,60.2,56.3,47.5,32.0.HRMS-ESI:m/z calculated for C ₂₅ H ₂₉ N ₆ O ₆ [M+H] ⁺ 509.2143,found509.2134.

Compound A57: 2-(4-(4-methoxyphenyl))-6-(4-nitrophenyl)pyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

Light yellow solid; total yield 15%; Mp: 259°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.25 (s, 1H), 9.70 (s, 1H), 9.02 (s, 2H) ,8.46-8.25(m,5H,aromatic),8.20-8.15(m,2H,aromatic),7.12-7.04(m,2H,aromatic),3.98-3.88(overlap,3H,tetrahydropyranyl),3.86(s,3H ₁₃ ^​ CNMR(100MHz,DMSO-d ₆ ,ppm)δ:165.2,162.5,162.3,157.4,152.9,149.0,141.3,129.3,128.6,127.1,123.9,114.4,108.4,65.4,55.6,47.5,31.9.HRMS- ESI :m/z calculated for C ₂₃ H ₂₅ N ₆ O ₄ [M+H] ⁺ 449.1932, found 449.1925.

Compound A58: 1-(4-hydroxyphenyl)-2-(4-(4-methoxyphenyl)-6-(4-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 16%; Mp: 238-240°C; ¹ H NMR (400 MHz, DMSO-d ₆ , ppm) δ: 11.36 (s, 1H), 9.95-9.74 (overlap, 2H), 8.77 (s, 2H), 8.38-8.21 (m, 4H, aromatic), 8.05-7.83 (m, 3H, aromatic), 7.36 (d, J＝8.3 Hz, 2H, aromatic), 6.99-6.82 (m, 4H, aromatic), 4.49 (d, J＝4.8 Hz, 2H, CH ₂ ), 3.84 (s, 3H, OCH ₃ ); ¹³ C NMR (100 MHz, DMSO-d ₆ ,ppm)δ:164.9,162.5,162.4,157.7,157.4,153.4,149.0,141.1,130.2,129.2,128.6,126.9,125.3,123.8,115.7,114.3,108.3,55.5,45.2.HRMS- ESI:m/z calculated for C ₂₅ H ₂₃ N ₆ O ₄ [M+H] ⁺ 471.1775,found471.1770.

Compound A59: 1-(2-hydroxyethyl)-2-(4-(4-methoxyphenyl)-6-(4-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 18%; Mp: 203-204°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.21 (s, 1H), 9.88 (s, 1H), 8.61 (s, 2H) ),8.50(d,J＝8.5Hz,2H,aromatic),8.38-8.29(m,3H,aromatic),8.27(d,J＝8.4Hz,2H,aromatic),7.08(d,J＝8.5Hz, 2H,aromatic),5.46(s,1H,OH),3.86(s,3H,OCH ₃ ),3.76-3.67(m,2H,hydroxyethyl,CH ₂ ),3.55-3.45(m,2H,hydroxyethyl,CH ₂ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm)δ:165.4,162.6,162.2,157.5,154.1,149.0,141.1,129.5,128.8,127.0,123.9,114.4,108.3,58.6,55.5,44.2.HRMS-ESI:m/zcalced for C ₂₀ H ₂₁ N ₆ O ₄ [M+H] ⁺ 409.1619, found 409.1615.

Compound A60: 2-(4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl))-6-(3-nitrophenyl)pyrimidin-2-yl)-1-(4-hydroxybenzyl)guanidine hydrochloride is characterized as follows:

White solid; total yield 23%; Mp: 259-260°C; ¹ H NMR (600MHz, DMSO-d ₆ , ppm) δ: 11.27 (s, 1H), 9.80-9.64 (overlap, 2H), 8.98-8.75 (overlap,3H),8.49(d,J＝7.6Hz,1H,aromatic),8.35(d,J＝9.7Hz,1H,aromatic),8.28(s,1H,aromatic),7.78-7.68(m,2H ,aromatic),7.36-7.27(m,3H,aromatic),6.88-6.75(m,3H,aromatic),4.51(d,J＝4.8Hz,2H,N-CH ₂ ),4.37-4.19(overlap,4H ,O-CH ₂ -CH ₂ -O); ¹³ C NMR (150MHz, DMSO-d ₆ , ppm) δ: 164.7, 162.5, 157.6, 157.3, 153.4, 148.3, 146.9, 143.7, 136.9, 133.5, 130.4, 129.8, 127.7, 125.9, 125.5, 121. 9 ,120.6,117.4,116.6,115.7,107.9,64.5,64.0,45.0.HRMS-ESI:m/zcalced for C ₂₆ H ₂₃ N ₆ O ₅ [M+H] ⁺ 499.1724, found 499.1717.

Compound A61: 2-(4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl))-6-(3-nitrophenyl)pyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 18%; Mp: 254-255°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.17 (s, 1H), 9.64 (d, J = 7.7Hz, 1H), 9.01(s,2H),8.83(s,1H,aromatic),8.64(d,J＝7.8Hz,1H,aromatic),8.36(d,J＝7.8Hz,1H,aromatic),8.26(s,1H, aromatic),7.81(td,J＝8.1,2.9Hz,1H,aromatic),7.70(d,J＝2.3Hz,2H,aromatic),7.02–6.88(m,1H,aromatic),4.38-4.25(overlap, 4H,O-CH ₂ -CH ₂ -O),4.02-3.80(overlap,3H,tetrahydropyranyl),3.52-3.39(overlap,2H,tetrahydropyranyl),2.09-1.92(m,2H,tetrahydropyranyl),1.67-1.48(m,2H,tetrahydropyranyl); ¹³ C NMR(100MHz,DMSO-d ₆ ,ppm)δ:164.9,162.3,157.2,152.9,148.3,147.0,143.7,136.9,133.8,130.5,127.9,125.9,121.6,121.0,117.5,116.3,107.9,65.4,64.5,64. 0,47.5,32.0.HRMS -ESI:m/z calculated for C ₂₄ H ₂₅ N ₆ O ₅ [M+H] ⁺ 477.1881,found 477.1873.

Compound A62: 2-(4-(4-methoxyphenyl))-6-(2-nitrophenyl)pyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 23%; Mp: 262-264°C; ¹ H NMR (400MHz, Methanol-d ₄ , ppm) δ: 8.09 (d, J = 8.9 Hz, 2H, aromatic), 7.98-7.94 (m, 1H, aromatic), 7.85 (s, 1H, aromatic), 7.82-7.75 (m, 2H, aromatic), 7.73-7.67 (m, 1H, aromatic), 7.01 (d, J = 8.9 Hz, 2H, aromatic), 3.89 (dt, J = 11.8, 3 .6Hz,2H,tetrahydropyranyl),3.84-3.73(overlap,4H),3.47(td,J＝11.5,2.2Hz,2H,tetrahydropyranyl),2.00-1.88(m,2H,tetrahydropyranyl),1.60(dtd,J＝12.9,10.8,4.3Hz,2H,tetrahydropyranyl) opyranyl); ¹³ CNMR (100MHz, Methanol-d ₄ , ppm) δ: 167.8, 165.7, 164.7, 158.3, 154.5, 150.4, 134.3, 133.2, 132.5, 132.5, 130.5, 129.4, 125.8, 115.6, 111.6, 67.1, 56.1,49.5,33.2.HRMS-ESI:m/zcalced for C ₂₃ H ₂₅ N ₆ O ₄ [M+H] ⁺ 449.1932, found 449.1923.

Compound A63: 1-(4-hydroxyphenyl)-2-(4-(4-methoxyphenyl)-6-(2-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 20%; Mp: 167-169°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.21 (s, 1H), 9.78 (overlap, 2H), 8.82 (s, 2H) ),8.15-8.02(m,2H,aromatic),7.96-7.84(m,4H,aromatic),7.81(t,J＝7.4Hz,1H,aromatic),7.29(d,J＝8.1Hz,2H,aromatic) ),6.97(d,J＝8.6Hz,2H,aromatic),6.84(d,J＝8.1Hz,2H,aromatic),4.48(d,J＝4.8Hz,2H,CH ₂ ),3.85(s,3H ,OCH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ ,ppm)δ:164.8,164.6,162.5,157.6,156.8,153.4,148.3,133.5,131.6,131.6,131.5,129.8,129.2,126.8,125.5,124.7,115.7, 114.5,110.4 ,55.6,45.1.HRMS-ESI:m/z calculated for C ₂₅ H ₂₃ N ₆ O ₄ [M+H] ⁺ 471.1775,found471.1770.

Compound A64: 1-(4-bromobenzyl)-2-(4-(4-methoxyphenyl)-6-(2-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 21%; Mp: 238-239°C; ¹ H NMR (400 MHz, DMSO-d ₆ , ppm) δ: 11.34 (s, 1H), 9.82 (s, 1H), 8.97 (s, 2H), 8.19-7.97 (m, 4H, aromatic), 7.97-7.77 (m, 3H, aromatic), 7.62 (d, J = 8.4 Hz, 2H, aromatic), 7.43 (d, J = 8.3 Hz, 2H, aromatic), 7.04 (d, J = 8.9 Hz, 2H, aromatic), 4.64 (d, J = 4.9 Hz, 2H, CH ₂ ), 3.87 (s, 3H, OCH ₃ ); ¹³ C NMR (100 MHz, DMSO-d ₆ ,ppm)δ:164.9,164.4,162.6,156.8,153.9,148.3,135.3,133.4,131.7,131.6,131.5,131.5,130.1,129.3,126.9,124.7,121.2,114.5,110.5,5 5.6,44.3.HRMS-ESI:m/zcalced for C ₂₅ H ₂₂ N ₆ O ₃ Br[M+H] ⁺ 533.0931, found 533.0927.

Compound A65: 2-(4,6-diphenylpyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 20%; Mp: 167-169°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.33 (s, 1H), 9.89 (d, J = 7.7Hz, 1H), 9.13(s,2H),8.33(s,1H,aromatic),8.24(d,J＝7.1Hz,4H,aromatic),7.70-7.48(m,6H,aromatic),4.06-3.84(overlap,3H,tetrahydropyranyl ),3.49-3.44(overlap,2H,tetrahydropyranyl),2.07-1.92(m,2H,tetrahydropyranyl),1.65-1.42(m,2H,tetrahydropyranyl); ¹³ CNMR(100MHz,DMSO-d ₆ ,ppm)δ:165.2,157.5,153.2,135.4,131.9,129.1,127.5,108.4,65.4,47.4,32.0.HRMS-ESI:m/z calculated forC ₂₂ H ₂₄ N ₅ O[M+H] ⁺ 374.1975, found 374.1971.

Compound A66: 1-(4-hydroxyphenyl)-2-(4-(4-methoxyphenyl)-6-(pyridin-2-yl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

White solid; total yield 22%; Mp: 188-189°C; ¹ H NMR (400MHz, DMSO-d ₆ , ppm) δ: 11.43 (s, 1H), 9.98 (s, 1H), 8.89 (s, 2H) ),8.74(dt,J＝4.8,1.3Hz,1H,aromatic),8.35(s,1H,aromatic),8.00-7.88(m,2H,aromatic),7.80(d,J＝8.4Hz,2H,aromatic) ),7.62(td,J＝4.9,3.3Hz,1H,aromatic),7.41-7.31(m,2H,aromatic),6.98-6.83(m,4H,aromatic),4.50(d,J＝4.5Hz,2H ,CH ₂ ), 3.83 (s, 3H, OCH ₃ ); ¹³ C NMR (100MHz, DMSO-d ₆ , ppm) δ: 164.7, 163.0, 162.3, 157.7, 157.4, 153.4, 151.5, 149.3, 138.3, 130.3, 128.9, 126.9 ,126.5,125.4,121.7,115.8,114.5,107.0,55.5,45.3.HRMS-ESI:m/z calculated for C ₂₄ H ₂₃ N ₆ O ₂ [M+H] ⁺ 427.1877, found 427.1881.

Compound A67: 2-(4-(4-methoxyphenyl))-6-(pyridin-2-yl)pyrimidin-2-yl)-1-(tetrahydro-2H-pyran-4-yl)guanidine hydrochloride was characterized as follows:

White solid; total yield 20%; Mp: 198-200℃; ¹ H NMR (400MHz, Methanol-d ₄ ,ppm)δ:8.84-8.80(m,2H,aromatic),8.64(t,J＝7.6Hz,1H,aromatic),8.44(s,1H,aromatic),8.14-7.99(m,3H,aromatic),7.00(d,J＝8.3Hz,2H,aromatic),3.96(dt,J＝12.0,3.4Hz, ^{2 13} C NMR (100MHz, Methanol-d ₄ ,ppm)δ:168.3,165.1,158.8,158.7,154.4,148.4,147.3,145.5,130.8,130.7,130.0,129.4,128.0,126.9,115.9,110.5,67.1,56.3,49.6,33.4 .HRMS-ESI:m/z calculated for C ₂₂ H ₂₅ N ₆ O ₂ [M+H] ⁺ 405.2034, found 405.2039.

Compound A68: 2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine hydrochloride is characterized as follows:

The preparation method of compound A68 is the same as that of compound A07, except that 4-nitrobenzylamine is used instead of 4-aminotetrahydropyran. When the benzoyl group of intermediate 6 is hydrolyzed and removed, the benzyl group on guanidine is also removed to obtain 2-(4-(4-methoxyphenyl))-6-(3-nitrophenyl)pyrimidin-2-yl)guanidine. Subsequently, the hydrochloride is prepared by the same method to obtain compound A68. Yellow solid; total yield 13%; Mp: 126-127°C; ¹ H NMR (400 MHz, DMSO-d ₆ , ppm) δ: 11.32 (s, 1H), 8.96 (s, 1H), 8.70 (d, J = 7.8 Hz, 1H, aromatic), 8.66-8.45 (overlap, 4H), 8.41-8.33 (overlap, 2H, aromatic), 8.25 (d, J = 8.6 Hz, 2H, aromatic), 7.84 (t, J = 8.0 Hz, 1H, aromatic), 7.08 (d, J = 8.7 Hz, 2H, aromatic), 3.85 (s, 3H, OCH ₃ ); ¹³ C NMR (100 MHz, DMSO-d ₆ ,ppm)δ:165.3,162.6,162.5,157.4,155.5,148.4,137.1,133.8,130.6,129.5,127.2,125.9,121.9,114.4,108.0,55.6.HRMS-ESI:m/z calculated for C ₁₈ H ₁₇ N ₆ O ₃ [M+H] ⁺ 365.1357,found 365.1349.

Test example

The compounds A01-A68 provided in Examples 1-68 were used to test their inhibitory effects on histone demethylase. The test method is as follows:

GSK-J1 was used as a reference compound for the KDM6B enzyme inhibition activity test; pyridine-2,4-dicarboxylic acid (2,4-PDCA) was used as a reference compound for the other KDM subfamily enzyme inhibition activity tests. The initial screening was performed by testing the inhibition rate of replicates at a single compound concentration (50 μM). For the calculation of IC ₅₀ , 5 concentrations of each compound were tested, starting at 50 μM and with a gradient of 5-fold dilutions. Substrate solution was prepared by adding peptide, ascorbic acid, 2-OG, and Fe(II) in 1x assay buffer. 5 μL of enzyme solution was transferred to the assay plate of the compound solution and incubated at room temperature for 15 minutes. 5 μL of substrate solution was added to each well and incubated at room temperature for 60 minutes. 15 μL of acceptor solution and donor solution were added to 1xAlphalisa buffer, incubated at room temperature under low light for 60 minutes, and the endpoint was read using EnSpire in Alpha mode. Curve fitting calculation, the inhibition rate was calculated by formula (1): Inh% = (Max-Signal) / (Max-Min) * 100. The IC ₅₀ value was obtained by fitting the data in GraphPad by formula (2): Y = Bottom + (Top-Bottom) / (1 + 10 ^∧ ((LogIC ₅₀ -X) * HillSlope)). (Where Y is the inhibition rate%, X is the compound concentration). The test results are shown in Table 2.

Table 2. Inhibitory activity of some compounds on KDM6B enzyme

50 compounds were selected for KDM6B enzyme inhibitory activity test. The results in Table 1 show that 30 compounds have a certain degree of inhibitory activity against KDM6B enzyme (IC ₅₀ <20 μM), among which 8 compounds A10, A24, A57-A61, and A68 have significant inhibitory activity (IC ₅₀ values are in the range of 1.22-2.90 μM).

Table 3. Inhibitory activity of some compounds on other subfamilies of KDM

[a]ND, no data available.

Some compounds with good activity were selected to test the inhibitory activity against other subtypes of JMJD KDM enzymes, and the results are shown in Table 2. Some compounds also have a certain degree of inhibitory effect on other subfamilies of KDM. Among them, compounds A07, A10, A11, A22, and A57 selectively inhibited KDM6B enzyme in the KDM6 subfamily, showing a selectivity superior to GSK-J1. This type of compound shows good development prospects.

In summary, the embodiments of the present invention provide a 4,6-diaryl-2-substituted pyrimidine guanidine compound and its preparation method and application, the compound has a novel structure, has a certain inhibitory effect on histone demethylase, and belongs to a new type of inhibitor structure. In addition, the preparation method of the compound is simple, easy to operate, has low requirements for equipment, and is suitable for large-scale industrial production in the later stage. The compound undergoes structural modification and in-depth pharmacological research, and has good application prospects in the preparation of histone demethylase inhibitors.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims (10)

1. A4, 6-diaryl-2-substituted pyrimidine guanidine compound is characterized in that the structural formula is

Wherein, R ₁ is selected from hydrogen, C1-C9 alkyl, C1-C9 substituted alkyl, C2-C9 alkenyl, C2-C9 substituted alkenyl, C2-C9 alkynyl, C2-C9 substituted alkynyl, C1-C9 alkoxy, C1-C9 substituted alkoxy, C5-C10 cycloalkyl, C5-C10 heterocycloalkyl, C6-C10 aryl or C6-C10 substituted aryl;

R ₂ and R ₃ are the same or different and are each independently selected from C6-C10 aryl or C6-C10 substituted aryl.

2. A process for the preparation of 4, 6-diaryl-2-substituted pyrimidine guanidine compounds as claimed in claim 1, comprising:

carrying out substitution reaction on the compound I and substituted amine to obtain a compound II;

The compound II is hydrolyzed under the action of an alkali reagent to obtain the 4, 6-diaryl-2-substituted pyrimidine guanidine compound;

wherein the structural formula of the compound I is The structural formula of the substituted amine is R ₄NH₂, and the structural formula of the compound II is

When R ₁ is hydrogen, the substituted amine is p-nitrobenzylamine and when the compound II is hydrolyzed, R ₄ is hydrolyzed to hydrogen; when R ₁ is not hydrogen, R ₄ and R ₁ are the same group.

3. The preparation method according to claim 2, wherein the molar ratio of the compound I to the substituted amine is 1:2 to 2.5.

4. A process according to claim 3, wherein the reaction of compound I with the substituted amine is carried out under the catalysis of EDCI and DIPEA, the molar ratio of compound I, EDCI and DIPEA being 1:1.8 to 2.2:1.8 to 2.2.

5. The process of claim 2 wherein compound I is obtained by reacting compound III with benzoyl isothiocyanate, wherein compound III has the formula

6. The method of claim 5, wherein the molar ratio of compound III to benzoyl isothiocyanate is 1: 3-5, the reaction temperature is 70-80 ℃ and the reaction time is 5-8 h.

7. The process according to claim 5, wherein the compound III is obtained by a condensation reaction of a compound IV with guanidine hydrochloride under the catalysis of a base, wherein the compound IV has the formula

8. The method of claim 6, wherein the compound IV and guanidine hydrochloride are used in an amount of 1:2.5 to 3; the reaction temperature is 70-80 ℃ and the reaction time is 7-10 h.

9. Use of a 4, 6-diaryl-2-substituted pyrimidine guanidine compound as claimed in claim 1 for preparing histone demethylase inhibitors.

10. The use according to claim 9, wherein the histone demethylase inhibitor comprises the 4, 6-diaryl-2-substituted pyrimidine guanidine compound, a salt thereof or a derivative thereof.