CN111995585B - Pyrimidine acetamides and their use as inhibitors of phosphodiesterase PDE2 activity - Google Patents

Pyrimidine acetamides and their use as inhibitors of phosphodiesterase PDE2 activity Download PDF

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CN111995585B
CN111995585B CN202010771191.7A CN202010771191A CN111995585B CN 111995585 B CN111995585 B CN 111995585B CN 202010771191 A CN202010771191 A CN 202010771191A CN 111995585 B CN111995585 B CN 111995585B
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pde2
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pde
acetamide
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CN111995585A (en
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冯筱晴
唐龙
黄险峰
孔韧
宋国强
高英
夏颜
谈颖
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Changzhou University
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/56—One oxygen atom and one sulfur atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/04—Centrally acting analgesics, e.g. opioids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/22—Anxiolytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/30—Drugs for disorders of the nervous system for treating abuse or dependence
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links

Abstract

The invention belongs to the technical field of medicines, and particularly discloses a pyrimidine acetamide compound and application thereof as a phosphodiesterase PDE2 activity inhibitor. At least one of pyrimidine acetamide compounds or hydrates, pharmaceutically acceptable salts, tautomers, stereoisomers and precursor compounds thereof is used as an active ingredient for preparing a medicament for preventing or treating PDE2 disorder. The pyrimidine acetamide compound can effectively inhibit the activity of PDE2 enzyme
Figure DDA0002616653080000011
kit and PDE-GloTMPhosphodiesterase Assay for determining the enzymatic level IC50Has good PDE2 enzyme inhibition effect. The compound of the invention is used as an effective inhibitor of PDE2, is expected to be used as an active ingredient for preparing a medicament for treating PDE2 disorder, and has medicinal prospect.

Description

Pyrimidine acetamides and their use as inhibitors of phosphodiesterase PDE2 activity
Technical Field
The invention belongs to the technical field of medicines, and particularly discloses a pyrimidine acetamide compound and application thereof as a phosphodiesterase PDE2 activity inhibitor.
Background
Adenosine cyclophosphate (Cyclic adenosine 3 ', 5' monophosphosphate, cAMP) was first discovered by all and Sutherland et al in 1958; guanosine monophosphate (cGMP) was discovered by Ashman et al in 1963. Since then, studies on cyclic adenosine monophosphate and cyclic guanosine monophosphate have received increasing attention.
Cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) regulate gene expression in cells by means of controlling ion channels, are important second messengers in cells and are involved in the realization of numerous physiological functions, such as learning memory, cell cycle regulation, cell differentiation, proliferation, inflammatory processes, smooth muscle contraction relaxation, visual signal transduction, and metabolic functions, such as steroid synthesis, insulin secretion, glycogen synthesis, lipogenesis, etc.
Intracellular levels of cAMP and cGMP are controlled by the corresponding cyclases and Phosphodiesterases (PDEs) to maintain their concentrations within an optimal range for responding to a signal. Adenylate cyclase and guanylate cyclase respectively catalyze ATP and GTP in vivo to be converted into cAMP and cGMP, and the concentration of a second messenger is increased; while PDEs are the only family of enzymes in the body that can hydrolyze cAMP and cGMP to the inactive substances 5 '-AMP and 5' -GMP, reducing the intracellular concentration of the second messenger. Therefore, PDE can be used as target of action of various disease drugs, and the effect of disease treatment is achieved by blocking hydrolysis process of PDE on cAMP/cGMP, maintaining intracellular concentration of second messenger, and reversing reduction of concentration in the disease process.
Cyclic nucleoside phosphodiesterases contain at least 11 structurally related but functionally distinct gene families (PDE1-PDE 11). High levels of PDE will result in the induction of visual deterioration, congestive heart failure, depression, asthma, erectile dysfunction and inflammation. Selective PDE inhibitors play an important role in inhibiting platelet aggregation, treating depression, parkinson's disease and learning disorders. Each family has 21 gene products with different isoforms and splice variants, and encodes these 11 PDE protein families. Examples of the PDE1 include PDE1A, PDE1B, PDE1C and the like.
PDE2(Phosphodiesterase2) is a member of the PDEs isozyme family, has only one subtype (PDE2A), is a homodimer structure, and has a dual action of hydrolyzing cAMP and cGMP simultaneously. PDE2 is mainly distributed in the central nervous system of human body and is expressed more in cortex of brain and hippocampus, and studies have shown that PDE2 can be involved in the regulation of central nervous system diseases, and corresponding PDE2 inhibitors have been shown to treat central nervous system diseases related to depression, anxiety and learning and memory disorders. The PDE2 inhibitor can improve mild cognitive impairment and age-related memory disorder diseases by inhibiting PDE2 protein activity to increase cAMP and cGMP levels, and inhibit thrombosis, and can be used for treating cancer, dementia and blood disorders. The existing PDE2 inhibitors are mainly EHNA, BAY60-7750, dipyridamole and the like, but are not yet on the market. It would therefore be desirable to find novel PDE2 inhibitors that would aid in later drug development.
Disclosure of Invention
The present invention provides compounds of formula I, and methods of using these compounds to treat PDE2 disorders by inhibiting PDE 2.
The present invention provides compounds of formula I and their docking forms, chelates, non-covalent complexes, prodrugs, stereoisomers, solvates, pharmaceutically acceptable salts and mixtures thereof.
Figure BDA0002616653060000031
X is NH-R1。
R1Selected from hydrogen, alkyl, aryl, heteroaryl, -C (═ O) -alkyl, -C (═ O) -aryl, or-C (═ O) -heteroaryl, any of which may optionally be substituted with one or more independent Q1Substituted by groups;
Q1selected from hydrogen, halogen, -CN, -CF3，-OCF3，-NO2Oxy, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkylAryl, heteroaryl, heterocycloaryl, -OR2，-S(O)nR3，-NR4R5，-SO2NR4R5，-C(O)R6，-C(O)NR4R5，-C(O)OR7，-OC(O)R8，-NR4C(O)R6，-NR4S(O)2R9，-NR10C(O)NR4R5，-NR10S(O)2NR4R5or-NR10S(O)NR4R5Any of the foregoing groups may optionally be substituted with one or more independent hydrogen, halogen, -CN, -OH, -NH2，-NO2Oxy, -CF3，-OCF3，-CO2H，-S(O)nH, alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, heteroaryl or-O-alkyl, any of which may be partially or fully halogenated;
n is 0, 1, or 2.
Further, the compound is 4- {2- [5- (2-Hydroxy-ethyl) -4-methyl-6-oxo-1,6-dihydro-pyrimidin-2-ylsulfanyl ] -acetylamino } -benzoic acid isopropyl ester (4- {2- [5- (2-Hydroxy-ethyl) -4-methyl-6-oxo-1,6-dihydro-pyrimidin-2-ylsulfanyl ] -acetylamino } -benzoic acid isopropyl ester) having the formula,
Figure BDA0002616653060000041
n- (3-Ethynyl-5,6-dihydro-4H-cyclopenta [ b ] thiophen-2-yl) -2- [5- (2-hydroxy-ethyl) -4-methyl-6-oxo-1, 6-dihydropyrimidin-2-ylsulfanyl ] -acetamide (N- (3-ethyl-5, 6-dihydro-4H-cyclopenta [ b ] thiophen-2-yl) -2- [5- (2-
hydroxy-ethyl)-4-methyl-6-oxo-1,6-dihydro-pyrimidin-2-ylsulfanyl]-acetamide) having the formula,
Figure BDA0002616653060000042
n- (4-fluorophenyl) -2- ((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)) Thio) acetamide (N- (4-fluorophenyl) -2- ((5- (2-hydroxyethenyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide), which has the structural formula shown in the specification,
Figure BDA0002616653060000043
n-benzyl-2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide (N-benzyl-2- ((5- (2-hydroxyethyi) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide), which has the structural formula
Figure BDA0002616653060000051
2- ((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) -N- (4-methoxybenzyl) acetamide (2- ((5- (2-hydroxyethyi) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) -N- (4-methoxybenzyl) acetamide), the structural formula of which is shown in the specification
Figure BDA0002616653060000052
N-butyl-2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide
(N-butyl-2- ((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide), the structural formula of which is shown in the specification
Figure BDA0002616653060000053
2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) -N- (1,3,4-thiadiazol-2-yl) acetamide
(2- ((5- (2-hydroxyethenyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) -N- (1,3,4-thiadiazol-2-yl) acetamide), the structural formula of which is shown in the specification
Figure BDA0002616653060000061
The invention also provides a novel use of a compound of formula I for the manufacture of a medicament for the treatment of a PDE2 disorder, wherein the PDE2 disorder is a central nervous system disorder. Central nervous system disorders are: psychiatric disorders and conditions; anxiety disorders; movement disorders; a substance-related disorder; mood disorders; neurodegenerative disorders; pain; autistic disorder.
The invention also provides a medicament prepared from the compound of the formula I, a hydrate, a pharmaceutically acceptable salt, a tautomer, a stereoisomer or a precursor compound thereof and one or more pharmaceutically acceptable carriers.
Wherein the carrier comprises diluent, excipient, filler, binder, humectant, disintegrant, absorption enhancer, surfactant, adsorption carrier, lubricant, etc. conventional in pharmaceutical field.
The medicine can be made into various forms such as injection, tablet, powder, granule, pill, capsule, oral liquid, ointment, cream, etc. The medicaments in various dosage forms can be prepared according to the conventional method in the pharmaceutical field.
The medicine is used for treating PDE2 disorder by inhibiting phosphodiesterase PDE2 activity, and can be introduced into body such as muscle, intradermal, subcutaneous, intravenous, and mucosal tissue by injection, spray, nasal drop, eye drop, penetration, absorption, physical or chemical mediated method; or mixed or coated with other materials and introduced into body.
Searching and finding out that the compound of formula I can inhibit the activity of PDE2 enzyme from SPECS library by adopting a method integrating virtual screening and experimental determination, carrying out advanced evaluation on the effectiveness of drug molecules by adopting a computer simulation method, detecting and verifying the activity by adopting a biological experimental method, obtaining a series of effective PDE2 enzyme inhibitors, and carrying out virtual screening and experimental determination on the inhibitors
Figure BDA0002616653060000071
cAMP kit and PDE-GloTMPhosphodiesterase Assay for determining the enzymatic level IC50The series of compounds have good PDE2 enzyme inhibition effect, and the researches are very important for finally developing anti-PDE 2 medicaments and have important application prospects.
Drawings
FIG. 1 is a drawing of
Figure BDA0002616653060000072
cAMP kit assay of the Compound 4- {2- [5- (2-hydroxy-ethyl) -4-methyl-6-oxo-1,6-dihydro-pyrimidin-2-ylsulfanyl]-a graph of IC50 values of isopropyl acetylamino } -benzoate at the PDE2 protein level;
FIG. 2 is PDE-GloTMPhosphodiesterase Assay of the Compound 4- {2- [5- (2-hydroxy-ethyl) -4-methyl-6-oxo-1,6-dihydro-pyrimidin-2-ylsulfanyl]-a graph of IC50 values of isopropyl acetylamino } -benzoate at the PDE2 protein level;
FIG. 3 is PDE-GloTMPhosphodienesterase Assay of N- (3-ethynyl-5,6-dihydro-4H-cyclopenta [ b ] b]Thien-2-yl) -2- [5- (2-hydroxy-ethyl) -4-methyl-6-oxo-1, 6-dihydropyrimidin-2-ylsulfanyl]IC of acetamide at the PDE2 protein level50A graph of values;
FIG. 4 is a drawing showing
Figure BDA0002616653060000073
A graph of the IC50 value of the compound N- (4-fluorophenyl) -2- ((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide at the PDE2 protein level as determined by AMP kit;
FIG. 5 is PDE-GloTMA graph of the IC50 value of compound N- (4-fluorophenyl) -2- ((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide at the PDE2 protein level, as measured by Phosphodiesterase Assay;
FIG. 6 is a drawing showing
Figure BDA0002616653060000074
A graph of the IC50 value of the compound N-benzyl-2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide at the PDE2 protein level as measured by AMP kit;
FIG. 7 is PDE-GloTMA graph of the IC50 value of the compound N-benzyl-2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide at the PDE2 protein level, as measured by Phosphodiesterase Assay;
FIG. 8 is a drawing showing
Figure BDA0002616653060000081
AMP kit determination of Compound 2- ((5- (2-hydroxyethyl)IC50 plot of yl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) -N- (4-methoxybenzyl) acetamide at the PDE2 protein level;
FIG. 9 is PDE-GloTMA graph of the IC50 value of the compound 2- ((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) -N- (4-methoxybenzyl) acetamide at the PDE2 protein level as measured by Phosphodiesterase Assay;
FIG. 10 is PDE-GloTMA graph of the IC50 value of the compound N-butyl-2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide at the PDE2 protein level, as measured by Phosphodiesterase Assay;
FIG. 11 is a schematic view of
Figure BDA0002616653060000082
AMP kit measured the compound 2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) -N- (1,3,4-thiadiazol-2-yl) acetamide IC50 value map at PDE2 protein level;
FIG. 12 is PDE-GloTMA graph of the IC50 value of the compound 2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) -N- (1,3,4-thiadiazol-2-yl) acetamide at the PDE2 protein level was determined by Phosphodiesterase Assay.
Detailed Description
The present invention is further described below with reference to examples, but is not limited thereto.
1. Experimental methods
1.1 virtual receptor-based screening
First adopt
Figure BDA0002616653060000083
The Protein Preparation module in the software package was used to manipulate the crystal structure of PDE2 (PDB: 4 HTX). The library of SPECS compounds (http:// www.specs.net /) was pretreated with ligaprep for compounds including removal of heavies, salt ions and minerals, possible ionization states and tautomers of the compounds at pH 7.4, and ring conformations, etc.
The molecular docking method is adopted to carry out virtual screeningBefore selection, the effectiveness of the adopted Glide docking method needs to be verified, the active site of PDE2 is defined, the center of mass of ligand molecules BAY60-7550 in a crystal structure is taken as the center, and the setting is carried out
Figure BDA0002616653060000091
In the cube region, the ligand molecule BAY60-7550 was treated with LigPrep in the same manner as in SP (Standard precision) parameter setting of Glide software, and then docked into the active pocket of PDE2 again, and Glide was found to better reproduce the binding conformation in the crystal structure.
The SPECS library compounds were docked and scored using the Glide SP (Standard Precision) mode, retaining the 3 ten thousand small molecule binding mode that precedes the score. In the crystal structure of the binding of BAY60-7550 to PDE2, hydrogen bond interactions with the active region residue GLN859 and pi-pi interactions with PHE862 are formed. For 3 ten thousand small molecule binding modes generated by docking, hydrogen bond standards are adopted as screening conditions, binding modes which form more than 1 hydrogen bond with GLN859 are selected, and 3660 qualified binding modes are obtained. In order to fully consider the structural diversity of the compounds, a Canvas module in Schrodinger is adopted for carrying out cluster analysis, reasonable binding modes are manually selected, 48 compounds are finally obtained, Shanghai ceramic company is entrusted as a proxy to purchase compound entities from SPECS company, and experimental determination is carried out. The compound with the structure shown in the formula I is found to have better PDE2 enzyme inhibition activity.
1.2 PDE2 in vitro enzyme Activity detection assay procedure
1) Expression and purification of PDE2 protein: pET15b-PDE2A (580-941) wild-type and truncated plasmids were awarded by the professor Cazechun university, North Ka, USA. After sequencing, the cells were transformed into E.coli BL21+ (Codon Plus) and induced to express (after induction with 0.1mM IPTG, incubation was continued at 15 ℃ for 20 hours), and then purified by Ni column to obtain His-tagged PDE2 protein.
2) PDE2 in vitro enzyme activity detection procedure:
use
Figure BDA0002616653060000101
cAMP kit assay inhibitory effect of compounds on PED 2:
add 4 μ LPDE2 protein and 2 μ L candidate compound (7 concentrations per compound, 3 parallel experiments) and incubate for 30 min;
then adding 4. mu.L Bio-cAMP to react with the residual PDE2 protein in the previous step for 1 hour;
adding 15 mu L of mixed solution of Donor Bead and Acceptor in a dark environment for reaction for 1 hour;
and reading the signal value of the micropore plate by using a multifunctional microplate reader.
Each group of experiments is provided with a positive maximum value control and a negative minimum value control, only a Bio-cAMP substrate is added without adding protein to be used as the positive control, and the substrate is the most at the moment, the signal value is the largest, and the control can be also called as a full inhibition control; protein and Bio-cAMP substrates were added as negative controls, protein was all involved in hydrolysis, and excess substrate gave the signal when the signal value was minimal, also referred to as the perhydrolysis control. Each of the above experiments was performed in 3 parallel experiments, each step requiring centrifugation (rotation speed 1000r/min, centrifugation for 1 minute).
② use of PDE-GloTMPhosphodiesterase Assay compounds were tested for their inhibitory effect on PDE 2:
mu.L of the compound to be screened (7 concentrations of each compound determined, 3 parallel experiments) and 1.5. mu.L of PDE2 were added to each well and incubated for 30 minutes at room temperature;
2.5. mu.L of 2. mu.M cAMP was added and incubated at room temperature for 20 minutes;
add 2.5. mu.L PDE-Glo per wellTMTermination Buffer，2.5μL PDE-GloTMDetection Solution, incubation for 20 minutes at room temperature;
finally adding 10 μ L of a solution containing
Figure BDA0002616653060000102
Of substrates
Figure BDA0002616653060000103
Buffer, incubation for 10 minutes at room temperature;
and reading the signal value of the micropore plate by using a multifunctional microplate reader.
Negative and positive controls were set for each set of experiments, negative controls: cAMP and no PDE protein were added. The highest cAMP content, highest PKA activity, and most ATP consumed by phosphorylated substrates, and therefore the weakest luminescent signal, is obtained with the lowest plate reading value without addition of PDE protein. Positive control: cAMP and PDE proteins were added. PDE hydrolyzes most of cAMP, PKA activity is reduced, ATP consumed by phosphorylated substrates is reduced, remaining ATP is increased, and luminescence signal is strongest, which is the set of the maximum plate reading value.
Example 1
With 4- {2- [5- (2-hydroxy-ethyl) -4-methyl-6-oxo-1,6-dihydro-pyrimidin-2-ylsulfanyl]-acetylamino } -benzoic acid isopropyl ester (SPECS library number: AS-871/43478043) via
Figure BDA0002616653060000111
AMP kit for detecting its enzymatic level IC50The value was 57.75. + -. 6.91. mu. mol/L (as shown in FIG. 1), which was in turn determined by PDE-GloTMPhosphodiesterase Assay for determining the enzymatic level IC50The value is 16.29 +/-0.35 mu mol/L (shown in figure 2), and the PDE2 enzyme inhibition effect is good;
example 2
Using N- (3-ethynyl-5,6-dihydro-4H-cyclopenta [ b ]]Thien-2-yl) -2- [5- (2-hydroxy-ethyl) -4-methyl-6-oxo-1, 6-dihydropyrimidin-2-ylsulfanyl]Acetamide (SPECS library number: AS-871/43478035) via PDE-GloTMPhosphodiesterase Assay for determining the enzymatic level IC50The value is 1.652 +/-0.352 mu mol/L (shown in figure 3), and the product has good PDE2 enzyme inhibition effect.
Example 3
N- (4-fluorophenyl) -2- ((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide via
Figure BDA0002616653060000112
AMP kit for detecting its enzymatic level IC50The value was 16.905. + -. 2.155. mu. mol/L (as shown in FIG. 4), which was in turn PDE-GloTMPhosphodiesterase Assay for determining the enzymatic level IC50The value was 6.068. + -. 0.872. mu. mol/L (as shown in FIG. 5), which was excellent in PDE2 enzyme inhibition effect.
Example 4
N-benzyl-2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide via
Figure BDA0002616653060000113
AMP kit for detecting its enzymatic level IC50The value was 34.67. + -. 4.58. mu. mol/L (as shown in FIG. 6), which was in turn PDE-GloTMPhosphodiesterase Assay for determining the enzymatic level IC50The value is 24.615 +/-3.705 mu mol/L (shown in figure 7), and the PDE2 enzyme inhibition effect is good.
Example 5
2- ((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) -N- (4-methoxybenzyl) acetamide via
Figure BDA0002616653060000121
AMP kit for detecting its enzymatic level IC50The value was 22.21. + -. 2.3. mu. mol/L (as shown in FIG. 8), which was in turn determined by PDE-GloTMPhosphodiesterase Assay for determining the enzymatic level IC50The value was 17.08. + -. 1.4. mu. mol/L (as shown in FIG. 9), which was excellent in PDE2 enzyme inhibitory effect.
Example 6
N-butyl-2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) acetamide via PDE-GloTMPhosphodiesterase Assay for determining the enzymatic level IC50The value was 38.79 + -6.17. mu. mol/L (as shown in FIG. 10), which showed good PDE2 enzyme inhibition effect.
Example 7
2- (((5- (2-hydroxyethyl) -4-methyl-6-oxo-1,6-dihydropyrimidin-2-yl) thio) -N- (1,3,4-thiadiazol-2-yl) acetamide via
Figure BDA0002616653060000122
AMP kit for detecting its enzymatic level IC50The value was 16.53. + -. 1.11. mu. mol/L (as shown in FIG. 11), which was in turn determined by PDE-GloTMPhosphodiesterase Assay for determining the enzymatic level IC50The value is 14.64 +/-1.38 mu mol/L (as shown in figure 12), and the PDE2 enzyme inhibition effect is good.
The present invention is not limited to the above-described embodiments, and any obvious improvements, substitutions or modifications can be made by those skilled in the art without departing from the spirit of the present invention.

Claims (6)

1. A pyrimidine acetamide compound and pharmaceutically acceptable salts thereof have the following structure,
Figure DEST_PATH_IMAGE002
、
Figure DEST_PATH_IMAGE004
or
Figure DEST_PATH_IMAGE006
。
2. A pharmaceutical composition made from a therapeutically effective amount of a compound, pharmaceutically acceptable salt, and one or more pharmaceutically acceptable carriers as claimed in claim 1.
3. The pharmaceutical composition of claim 2, wherein the carrier is selected from the group consisting of diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorptive carriers, and lubricants, which are conventional in the pharmaceutical field.
4. The pharmaceutical composition of claim 2, wherein the dosage form of the drug is tablet, capsule, granule, pill or other conventional dosage forms capable of being prepared.
5. Use of a compound according to claim 1 and pharmaceutically acceptable salts thereof for the manufacture of a medicament for the treatment of phosphodiesterase PDE2 disorder.
6. The use of a compound according to claim 5, wherein the PDE2 disorder is a central nervous system disorder.
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