Pyrrole [2, 3-d] pyrimidine compound or pharmaceutically acceptable salt and application thereof

Abstract

The invention discloses a pyrrole [2, 3-d] pyrimidine compound or a pharmaceutically acceptable salt of the pyrrole [2, 3-d] pyrimidine compound. The pyrrole [2, 3-d] pyrimidine compound can show good inhibitory activity on phosphodiesterase 7, and can be used as an inhibitor of the phosphodiesterase 7. On the basis, the pyrrole [2, 3-d] pyrimidine compound has a relatively good treatment effect on diseases related to phosphodiesterase 7.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical chemistry, and more specifically, relates to pyrrole [2,3-d] pyrimidine compounds or pharmaceutically acceptable salts thereof and applications thereof. Background Art

[0002] Phosphodiesterases (PDEs) can specifically hydrolyze cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) to generate the corresponding AMP and / or GMP and control their concentration in cells, thus affecting a variety of physiological processes and metabolic functions. PDEs are composed of 11 families (PDE1-11), among which PDE4, PDE7 and PDE8 specifically recognize and hydrolyze cAMP, PDE5, PDE6 and PDE9 specifically recognize and hydrolyze cGMP, and PDE1, PDE2, PDE3, PDE10 and PDE11 can recognize and hydrolyze cAMP and cGMP. PDE7 affects downstream pathways by regulating cAMP levels, thereby affecting cell growth processes such as transcription, cell cycle progression, metabolism and apoptosis, and inducing certain diseases such as asthma, autoimmune diseases and neurodegenerative diseases.

[0003] PDE7 has two subtypes, PDE7A-B, which are widely expressed in the human body, such as the heart, skeletal muscle, spleen, brain, lung, pancreas, bladder, stomach, small intestine, etc. PDE7A is distributed in various types of human proinflammatory cells and immune cells, including T lymphocytes, CD4+ and CD8+, etc., and participates in the regulation of inflammatory responses. PDE7 inhibitors play an important role in reducing inflammation. They are often used in combination with PDE4 inhibitors to jointly regulate the level of cAMP, activate PKA, affect the downstream pathways that produce inflammatory factors, and inhibit excessive inflammation. They are often used in autoimmune diseases such as multiple sclerosis. Therefore, PDE7 is believed to play a role in T cell activation and proliferation, inflammation, and regulation of various physiological substances.

[0004] In addition, PDE7A and PDE7B are widely expressed in the central nervous system, but there are differences in their distribution in different brain regions. PDE7A is mainly distributed in the cerebral cortex, hypothalamus, hippocampus, striatum and thalamus, while PDE7B is mainly distributed in the caudate nucleus and nucleus accumbens. Central nervous system processes such as neurogenesis, synaptogenesis and the formation of long-term memory Increased expression and activity of PDE7 have been detected in a variety of disease states, including neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and Huntington's disease, autoimmune diseases such as multiple sclerosis and chronic obstructive pulmonary disease, and several types of cancer. Early studies have shown that the administration of PDE7 inhibitors may improve these clinical disease symptoms. Therefore, targeting PDE7 may provide a new therapeutic strategy for a wide range of diseases and provide a complementary alternative to inhibitors of other cAMP-selective PDEs (such as PDE4). It is well known that PDE4 inhibitors are severely limited by their side effects, while PDE7 inhibitors have no vomiting side effects at effective doses. However, no PDE7 inhibitors are currently on the market. Summary of the invention

[0005] In view of the above-mentioned prior art problems, the primary purpose of the present invention is to provide a pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof. The pyrrole [2,3-d] pyrimidine compound can exhibit good inhibitory activity against phosphodiesterase 7 and has a good therapeutic effect on phosphodiesterase 7-related diseases.

[0006] The second object of the present invention is to provide a pharmaceutical composition.

[0007] The third object of the present invention is to provide a use of a pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt or a pharmaceutical composition thereof in the preparation of a phosphodiesterase 7 inhibitor.

[0008] The fourth object of the present invention is to provide the use of pyrrole [2,3-d] pyrimidine compounds or pharmaceutically acceptable salts or pharmaceutical compositions thereof in the preparation of drugs for preventing and treating phosphodiesterase 7 related diseases.

[0009] A fifth object of the present invention is to provide a use of a pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition in the preparation of a drug for treating alcoholic hepatitis.

[0010] The above purpose of the present invention is achieved through the following technical solutions:

[0011] The present invention claims protection for a pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof, the structural formula of which is as shown in the following formula (I):

[0012]

[0013] Wherein, R1 is selected from C 2-8 Alkenyl, C 1-8 Alkyl, C 3-8 Cycloalkyl, aryl C 1-8 Alkyl, heteroaryl C 1-8 Alkyl, C 1-8 Alkyl carboxyl, C 3-8 Heterocyclic group; wherein the aryl C 1-8 Alkyl, heteroaryl C 1-8 Alkyl, C 3-8 The heterocyclic group is unsubstituted or substituted with one or more selected from C 1-8 Alkoxy, C 1-6 Alkyl ester group, C 2-8 Alkenyl, aryl C 1-8 Alkyl, C 1-8 Alkylcarbonyl, C 3-8 The substituents of cycloalkylcarbonyl and heteroarylcarbonyl are substituted; wherein C 1-8 Alkylcarbonyl, C 3-8 Cycloalkylcarbonyl and heteroarylcarbonyl are unsubstituted or substituted with one or more selected from C 1-8 Alkyl or cyano substituents;

[0014] R2 is selected from NR4R5; wherein R4 and R5 are each independently selected from hydrogen, C 3-8 Cycloalkyl, C 1-8 Alkyl, aryl, heteroaryl; wherein the C 3-8 Cycloalkyl, C 1-8 Alkyl, aryl, heteroaryl are unsubstituted or substituted with one or more substituents independently selected from halogen;

[0015] R3 is selected from C 1-8 alkyl.

[0016] Preferably, R1 is selected from C 2-6 Alkenyl, C 1-6 Alkyl, C 3-6 Cycloalkyl, aryl C 1-8 Alkyl, heteroaryl C 1-8 Alkyl, C 1-8 Alkyl carboxyl, C 3-8 Heterocyclic group; wherein the aryl C 1-8 The aryl group in the alkyl group is a 5-6 membered aromatic ring, and the heteroaryl C 1-8 The heteroaryl group in the alkyl group is a 5-6 membered aromatic heterocyclic ring;

[0017] Wherein, the 5-6 membered aromatic ring, 5-6 membered aromatic heterocyclic ring, C 3-8 The heterocyclic group is unsubstituted or substituted with one or more selected from C 1-5 Alkoxy, C 1-6 Alkyl ester group, C2-8 Alkenyl, benzyl, C 1-5 Alkylcarbonyl, C 3-6 Substituted by cycloalkylcarbonyl, pyridinecarbonyl; wherein C 1-5 Alkylcarbonyl, C 3-6 Cycloalkylcarbonyl and pyridinecarbonyl are unsubstituted or substituted with one or more selected from C 1-6 Alkyl or cyano substituents;

[0018] R2 is selected from NR4R5; wherein R4 and R5 are each independently selected from hydrogen, C 3-8 Cycloalkyl, C 1-5 alkyl, 5-6 membered aromatic ring, 5-6 membered aromatic heterocyclic ring; wherein the C 3-8 Cycloalkyl, C 1-5 Alkyl, 5-6 membered aromatic ring, 5-6 membered aromatic heterocyclic ring are unsubstituted or substituted with one or more substituents independently selected from halogen;

[0019] R3 is selected from C 1-5 alkyl.

[0020] Preferably, R1 is selected from C 2-6 Alkenyl, C 1-6 Alkyl, C 3-4 Cycloalkyl, aryl C 1-8 Alkyl, heteroaryl C 1-8 Alkyl, C 1-8 Alkyl carboxyl, C 3-8 Heterocyclic group; wherein the aryl C 1-8 The aryl group in the alkyl group is a 5-6 membered aromatic ring, and the heteroaryl C 1-8 The heteroaryl group in the alkyl group is a 5-6 membered aromatic heterocyclic ring;

[0021] Wherein, the 5-6 membered aromatic ring, 5-6 membered aromatic heterocyclic ring, C 3-8 The heterocyclic group is unsubstituted or substituted with one or more C 1-5 Alkoxy, C 1-6 Alkyl ester group, C 2-8 Alkenyl, benzyl, C 1-5 Alkylcarbonyl, C 3-6 cycloalkylcarbonyl, pyridinecarbonyl substituents; wherein the pyridinecarbonyl is unsubstituted or substituted with one or more selected from C 1-6 Substitution of alkyl groups;

[0022] R2 is selected from NR4R5; wherein R4 and R5 are each independently selected from hydrogen, C 3-5 Cycloalkyl, C 1-5 Alkyl, 5-6 membered aromatic ring;

[0023] R3 is selected from C 1-3 alkyl.

[0024] Preferably, R1 is selected from ethylene, propylene, butene, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl C 1-3 Alkyl, heteroaryl C 1-3 Alkyl, C 1-4 Alkylcarboxyl, aminopiperidinyl, azetidinyl;

[0025] Wherein, the heteroaryl C 1-3 The heteroaryl in the alkyl group is pyridyl, furyl, thienyl, pyrrolyl, thiazolyl, imidazolyl, pyrazinyl, pyrimidinyl; the phenyl, pyridyl, furyl, thienyl, pyrrolyl, thiazolyl, imidazolyl, pyrazinyl, pyrimidinyl, aminopiperidinyl, azetidinyl are unsubstituted or substituted with one or more substituents selected from methoxy, ethoxy, propoxy, tert-butyl ester, ethylene, propylene, butylene, benzyl, methylcarbonyl, ethylcarbonyl, propylcarbonyl, cyclopropylcarbonyl, pyridylcarbonyl; wherein cyclopropylcarbonyl and pyridylcarbonyl are unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl or cyano;

[0026] R2 is selected from NR4R5; wherein R4 and R5 are each independently selected from hydrogen, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, phenyl, pyridyl, furyl, thienyl, pyrrolyl, thiazolyl, imidazolyl, pyrazinyl, pyrimidinyl; wherein the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, pyridyl, furyl, thienyl, pyrrolyl, thiazolyl, imidazolyl, pyrazinyl, pyrimidinyl are unsubstituted or substituted with one or more substituents independently selected from halogen;

[0027] R3 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl.

[0028] Preferably, R1 is selected from

[0029] R2 is selected from hydrogen,

[0030] Further preferably, the pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof is selected from any of the following structures:

[0031]

[0032] Furthermore, the present invention seeks to protect a pharmaceutical composition comprising the above-mentioned pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable excipient.

[0033] Furthermore, the present invention seeks to protect the use of the above-mentioned pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition in the preparation of a phosphodiesterase 7 inhibitor.

[0034] Furthermore, the present invention seeks to protect the use of pyrrole [2,3-d] pyrimidine compounds or pharmaceutically acceptable salts thereof or the above pharmaceutical compositions in the preparation of drugs for preventing and treating phosphodiesterase 7-related diseases.

[0035] Preferably, the related diseases include one or more of inflammatory diseases, autoimmune diseases, addictive diseases, primary impulse control disorders, and neurodegenerative diseases.

[0036] Preferably, the inflammatory disease comprises one or more of hepatitis, rhinitis, bronchitis, rheumatoid arthritis, inflammatory bowel disease, or gout, fever and pain associated with inflammation.

[0037] In the present invention, rhinitis can be seasonal allergic rhinitis; or perennial allergic rhinitis; or sinusitis of any type, etiology or pathogenesis, or sinusitis selected from purulent or non-purulent sinusitis; acute or chronic sinusitis; and ethmoid, anterior, maxillary or sphenoid sinusitis.

[0038] In the present invention, bronchitis can be bronchitis of any type, etiology or pathogenesis; or bronchitis selected from acute bronchitis, acute laryngotracheobronchitis, peanut inhalation bronchitis, catarrhal bronchitis, croup bronchitis, dry bronchitis, infectious asthmatic bronchitis, hyperplastic bronchitis, bronchitis caused by Staphylococcus or Streptococcus, and alveolar bronchitis.

[0039] In the present invention, rheumatoid arthritis can be rheumatoid arthritis of any type, etiology or pathogenesis; or rheumatoid arthritis selected from acute arthritis, acute gouty arthritis, chronic inflammatory arthritis, degenerative arthritis, infectious arthritis, Lyme arthritis, proliferative arthritis, psoriatic arthritis and vertebral osteoarthritis.

[0040] In the present invention, inflammatory bowel disease can be inflammatory bowel disease (IBD) of any type, etiology or pathogenesis; or inflammatory bowel disease selected from ulcerative colitis (UC), collagenous colitis; or selected from polyposis colitis, transmural colitis and segmental enteritis (CD).

[0041] In the present invention, the autoimmune disease may be multiple sclerosis or chronic nonspecific inflammatory disease of the digestive tract (eg Crohn's disease).

[0042] Preferably, the multiple sclerosis may be of any type, etiology or pathogenesis, or may be selected from the group consisting of primary, progressive and relapsing-remitting multiple sclerosis.

[0043] In the present invention, the addictive disease may be one or more of alcohol addiction, drug addiction, binge eating addiction, and the like.

[0044] In the present invention, the neurodegenerative disease can be one or more of stroke, epilepsy, Alzheimer's disease, Parkinson's syndrome, and Huntington's disease.

[0045] Furthermore, use of a pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof or the above pharmaceutical composition in the preparation of a drug for treating alcoholic hepatitis.

[0046] Compared with the prior art, the present invention has the following beneficial effects:

[0047] The present invention provides a pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof. The pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof exhibits good inhibitory activity against PDE7 and has good development prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 This is a schematic diagram of the changes in body weight of mice in the control group, alcohol group and drug-treated group in Example 23.

[0049] Figure 2 Schematic diagram of ALT levels in mouse plasma in the control group, alcohol group and drug-treated group in Example 23.

[0050] Figure 3 This is a schematic diagram of AST levels in mouse plasma in the control group, alcohol group and drug-treated group in Example 23.

[0051] Figure 4 This is a schematic diagram of TG levels in mouse plasma in the control group, alcohol group and drug-treated group in Example 23.

[0052] Figure 5 This is a schematic diagram of the IL-6 mRNA levels in the control group, alcohol group and drug-treated group in Example 23.

[0053] Figure 6 This is a schematic diagram of the IL-1β mRNA levels in the control group, alcohol group and drug-treated group in Example 23.

[0054] Figure 7 This is a schematic diagram of TNF-α mRNA levels in the control group, alcohol group and drug-treated group in Example 23.

[0055] Figure 8This is a schematic diagram of staining in the control group, alcohol group and drug-treated group in Example 23. DETAILED DESCRIPTION

[0056] The following examples are illustrative of the method of the present invention rather than limiting it. Other modifications and adaptive changes of various conditions and parameters that are obvious to those skilled in the art are all included in the spirit and scope of the present invention.

[0057] The term "halogen" refers to fluorine, chlorine, bromine and iodine. The term "halo" before a radical name means that the radical is partially or completely halogenated, ie substituted by F, Cl, Br or I in any combination.

[0058] The term "alkyl" refers to a saturated straight or branched hydrocarbon group. The alkyl group may preferably be a C1-C8 alkyl group, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl or the like.

[0059] The term "cycloalkyl" refers to a saturated or partially unsaturated cyclic hydrocarbon radical, including monocyclic or polycyclic groups. Unless otherwise specified, cycloalkyl has 3-12 carbon atoms. For example, cycloalkyl can be a monocyclic group with 3-12 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-3-enyl, cyclohexene, 1-cyclohex-1-enyl, cyclohexadiene, cycloheptene, cyclooctene, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl or similar groups. Cycloalkyl can also be a bicyclic group with 4-12 carbon atoms, such as [4,5], [5,5], [5,6] and [6,6] ring systems, bicyclic, selected from bicyclo [2.2.1] heptane, bicyclo [2.2.2] octane and bicyclo [3.2.2] nonane bridged bicyclic or similar groups. The cycloalkyl group may be saturated or have at least one double bond, but is not fully conjugated and is not aromatic.

[0060] The term "heterocyclyl" refers to a ring selected from the following: a 4-12 membered monocyclic, bicyclic saturated or partially unsaturated ring, which contains at least one carbon atom in addition to 1, 2, 3 or 4 heteroatoms selected from O, S and N. The heterocyclic ring may be saturated or contain at least one double bond. The heterocyclic ring may be oxo-substituted, i.e. substituted by the group =O. The heterocyclic group may be connected to the rest of the molecule via a carbon atom or a heteroatom. Examples of heterocyclic groups include, but are not limited to, tetrahydrofuranyl, azetidinyl, tetrahydropyranyl, piperidinyl, morpholinyl, piperazinyl, aminopiperidinyl, etc.

[0061] The term "alkoxy" refers to a saturated straight or branched group connected to the rest of the molecule through an oxygen atom, usually represented by RO-, where R is an alkyl group. Unless otherwise specified, the alkoxy group is an alkoxy group having 1 to 8 carbon atoms, examples of which include but are not limited to methoxy, ethoxy, etc.

[0062] The term "alkenyl" refers to a straight or branched hydrocarbon group containing at least one degree of unsaturation, wherein at least one position of CC is sp 2 Unless otherwise specified, the alkenyl group is an alkenyl group having 2 to 8 carbon atoms, and the alkenyl group includes "cis", "trans" or "Z", "E" isomers. Examples of alkenyl groups include ethylene, propylene, butene or similar groups.

[0063] The term "aryl" refers to a 5-6 membered aromatic ring, a 7-12 membered bicyclic ring system, wherein at least one ring in the bicyclic ring system is an aromatic ring. For example, (a) a 5-6 membered aromatic ring includes but is not limited to a phenyl group; (b) an aryl group of a 7-12 membered bicyclic ring system, such as a naphthyl group.

[0064] The term "heteroaryl" refers to a 5-6 membered aromatic ring, a 7-12 membered bicyclic ring system, wherein at least one ring in the bicyclic ring system is an aromatic ring, and the aromatic ring contains at least one heteroatom selected from O, S and N. For example, (a) a 5-6 membered heteroaryl includes, but is not limited to, a pyridine ring or a furan ring; (b) a heteroaryl of a 7-12 membered bicyclic ring system includes, but is not limited to, a benzodioxanyl group or a quinolinyl group.

[0065] The term "alkyl ester group" refers to the group -COOR, wherein R is an alkyl group. Unless otherwise specified, the alkyl ester group has 1 to 6 carbon atoms.

[0066] The term "alkylcarboxyl" refers to the group -RCOOH, wherein R is an alkyl group. Unless otherwise specified, the alkyl group in the alkyl ester group has 1 to 8 carbon atoms.

[0067] The terms "alkylcarbonyl", "cycloalkylcarbonyl" and "arylcarbonyl" refer to alkyl, cycloalkyl and aryl groups containing the group -C(=O)-, respectively, wherein the groups are connected to the rest of the molecule through the carbon atom on the carbonyl group. Unless otherwise specified, the alkyl, cycloalkyl and aryl groups have the meanings described in the present invention.

[0068] The term "acceptable salt" refers to pharmaceutically acceptable salt forms of these compounds obtained from various organic and inorganic acids and bases according to methods well known in the art. When the compound of formula (I) contains a carboxylic acid group, the compound can be reacted with a suitable base to form a suitable salt thereof to obtain the corresponding base addition salt. Examples of the base are alkali metal hydroxides, including potassium hydroxide, sodium hydroxide and lithium hydroxide; alkaline earth metal hydroxides, such as barium hydroxide and calcium hydroxide; alkali metal alcoholates, such as potassium ethoxide and sodium propoxide; and various organic bases, such as piperidine, diethanolamine and N-methylglutamide. The compound of formula (I) can also be reacted with a suitable acid to form a suitable salt thereof to obtain the corresponding acid addition salt. It can be, for example, a salt formed with hydrochloric acid, hydrobromic acid, phosphoric acid, nitrous acid, sulfuric acid, benzoic acid, citric acid, gluconic acid, lactic acid, maleic acid, succinic acid, tartaric acid, acetic acid, propionic acid, oxalic acid, maleic acid, fumaric acid, glutamic acid, pyroglutamic acid, salicylic acid, salicylic acid, saccharin and sulfonic acids (e.g., methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid and benzenesulfonic acid). Some of the acids listed above are dibasic or tribasic acids, i.e., acids containing two or three acidic hydrogens (e.g., phosphoric acid, sulfuric acid, fumaric acid and maleic acid).

[0069] The term "excipient" or "pharmaceutically acceptable excipient" refers to pharmaceutical excipients, including but not limited to fillers, anti-adhesives, binders, coatings, colorants, disintegrants, flavoring agents, glidants, lubricants, preservatives, sorbents, sweeteners, solvents, vehicles and adjuvants. For example, microcrystalline cellulose, corn starch, lactose, mannitol, povidone, croscarmellose sodium, sucrose, talc, gelatin, pectin, magnesium stearate, stearic acid, lower alkyl ethers of cellulose, ethanol, propylene glycol, glycerol, polyethylene glycol, poloxamer, sorbitol, polysorbate, monoglycerides and diglycerides, cyclodextrin, coconut oil, palm oil and water, etc.

[0070] In the present invention, the optically active forms (stereoisomers), enantiomers, racemates, diastereomers, hydrates and solvates of the compound of formula (I). The solvate of the compound refers to the inert solvent molecules adducted to the compound due to the mutual attraction between the inert solvent molecules and the compound. The solvate is, for example, a monohydrate, a dihydrate or an alcoholate.

[0071] Example 1

[0072]

[0073] Preparation of 7-allyl-2-(cyclopentylamino)-3-ethyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 1):

[0074] Synthesis route:

[0075]

[0076] Step 1: Synthesis of intermediate 1-1

[0077] 2,4-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (21mmol, 1.0equiv.) was dissolved in ethyl acetate (25mL), and triethylamine (63mmol, 1.0equiv.) and 4-dimethylaminopyridine (1.05mmol, 0.05equiv.) were added dropwise at room temperature. Then, di-tert-butyl dicarbonate (21mmol, 1.0equiv.) was added at 0°C, and the mixture was stirred at room temperature. The reaction was monitored by TLC, and the solvent was dried and recrystallized to obtain a white solid (yield 89%).

[0078] 1 HNMR (400MHz, CDCl3, ppm) δ7.67 (d, J = 4.0Hz, 1H), 6.61 (d, J = 3.9Hz, 1H), 1.65 (s, 9H). 13 CNMR (101MHz, CDCl3, ppm) δ154.18,153.20,152.71,146.50,128.12,118.57,102.46,86.34,27.83.

[0079] Step 2: Synthesis of intermediate 1-2

[0080] Compound 1-1 (18 mmol, 1.0 equiv.) was dissolved in 1,4-dioxane (25 mL) and heated to 80°C. An equal volume of 2 mol / L NaOH aqueous solution was added at 80°C and stirred for 4 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, 10% acetic acid was added, the pH was adjusted to about 6, and then extracted with DCM / CH3OH (v / v=10:1). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product can be directly used in the next step without purification.

[0081] Step 3: Synthesis of intermediate 1-3

[0082] The crude product obtained in the previous step, iodoethane (22.5mmol, 1.5equiv.) and potassium carbonate (22.5mmol, 1.5equiv.) were dissolved in NN-dimethylformamide (40mL) and stirred at room temperature for 5 hours. After the reaction was completed, it was diluted with water, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product was purified by rapid silica gel chromatography with ethyl acetate / petroleum ether = 1:5 (V / V) as the eluent to obtain a white solid (yield 61%).

[0083] 1HNMR (400MHz, CDCl3, ppm) δ7.30 (d, J = 3.9Hz, 1H), 6.65 (d, J = 3.9Hz, 1H), 4.37 (q, J = 7.1Hz, 2H), 1.65 (s, 9H), 1.37 (t, J = 7.1Hz, 3H). 13 CNMR (101MHz, CDCl3, ppm) δ158.05,147.03,146.08,145.13,122.51,109.81,104.93,85.29,41.90,27.84,13.47.

[0084] Step 4: Synthesis of intermediate 1-4

[0085] Compound 1-3 (9 mmol, 1.0 equiv.) was dissolved in DCM (15 mL) at room temperature, cooled to 0°C, and an equal volume of 4 mol / L hydrochloric acid / 1,4-dioxane solution was added. After stirring for 5 minutes, the mixture was moved to room temperature and stirred for another hour. TLC monitored the completion of the reaction, and the reaction mixture was concentrated under reduced pressure to obtain a crude salt product. Saturated potassium carbonate solution was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a white solid (yield 98%).

[0086] 1 HNMR (400MHz, DMSO-d6, ppm) δ12.02 (s, 1H), 7.09 (d, J = 3.4Hz, 1H), 6.49 (d, J = 3.3Hz, 1H), 4.23 (q, J = 7.1Hz, 2H), 1.26 (t, J = 7.0Hz, 3H). 13 CNMR (101MHz, CDCl3, ppm) δ159.09,145.34,143.69,121.17,106.32,103.95,42.33,13.70.

[0087] Step 5: Synthesis of Intermediate 1-5

[0088] Compound 1-4 (1mmol, 1.0equiv.), potassium carbonate (1.5mmol, 1.5equiv.), 3-bromopropylene (1.5mmol, 1.5equiv.) were dissolved in NN-dimethylformamide (5mL) and stirred at room temperature for 5 hours. After the reaction was completed, it was diluted with water, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product was purified by rapid silica gel chromatography with ethyl acetate / petroleum ether = 1:10 (V / V) as the eluent to obtain a white solid (yield 90%).

[0089] 1HNMR (400MHz, CDCl3, ppm) δ6.80 (d, J = 3.4Hz, 1H), 6.65 (d, J = 3.4Hz, 1H), 6.03–5.89 (m, 1H), 5.23 (dd, J = 10.2, 1.0Hz, 1H), 5.11 (dd, J=17.0, 1.0Hz, 1H), 4.69 (d, J=5.6Hz, 2H), 4.35 (q, J=7.1Hz, 2H), 1.36 (t, J=7.1Hz, 3H). 13 CNMR (101MHz, CDCl3, ppm) δ158.50,144.90,143.23,132.87,123.23,118.07,106.23,103.05,46.85,41.61,13.83.

[0090] Step 6: Synthesis of compound 1

[0091] Compound 1-5 (1mmol, 1.0equiv.) was dissolved in dimethyl sulfoxide (5mL), and cyclopentylamine (3mmol, 3.0equiv.) and N,N-diisopropylethylamine (3mmol, 3.0equiv.) were added under nitrogen protection at room temperature, and the temperature was raised to 80°C and stirred for 12 hours. After the reaction was completed, it was diluted with water, extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product was purified by rapid silica gel chromatography with ethyl acetate / petroleum ether = 1:2 (V / V) as the eluent to obtain a white solid (yield 73%)

[0092] 1 HNMR(400MHz, CDCl3, ppm) δ6.54(d,J=3.4Hz,1H),6.50(d,J=3.4Hz,1H),6.05–5.88(m,1H),5.24–5.08(m,2H),4.60(d,J=5.7Hz,2H), 4.40(s,1H),4.38–4.25(m,1H),4.04(q,J=7.2Hz,2H),2.19–2.06(m,2H),1.78–1.62(m,4H),1.55–1.43(m,2H),1.28(t,J=7.2Hz,3H) .13 CNMR (101MHz, CDCl3, ppm) δ159.06,149.99,148.06,133.85,119.78,117.31,102.21,100.25,53.87,46.42,35.27,33.32,23.84,13.07.

[0093] Example 2

[0094]

[0095] Preparation of 2-(cyclopentylamino)-3-ethyl-7-isopropyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 2):

[0096] The preparation process is the same as that in Example 1, except that 3-bromopropene in step 5 is replaced with 2-bromopropane to prepare 2-(cyclopentylamino)-3-ethyl-7-isopropyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 2).

[0097] 1 HNMR (400MHz, CDCl3, ppm) δ6.61 (d, J = 3.3Hz, 1H), 6.47 (d, J = 3.3Hz, 1H), 4.81–4.66 (m, 1H), 4.46 (s, 1H), 4.38–4.25 (m, 1H), 4.04(q,J=7.0Hz,2H),2.17–2.08(m,2H),1.77–1.63(m,4H),1.55–1.46(m,2H),1.43(d,J=6.8Hz,6H),1.26(t,J=7.2Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ159.04,149.60,147.44,116.45,101.72,100.40,53.81,45.73,35.15,33.26,23.84,22.58,13.07.

[0098] Example 3

[0099]

[0100] Preparation of 7-cyclopentyl-2-(cyclopentylamino)-3-ethyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 3):

[0101] The preparation process is the same as that in Example 1, except that 3-bromopropylene in step 5 is replaced with bromocyclopentane to prepare 7-cyclopentyl-2-(cyclopentylamino)-3-ethyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 3).

[0102] 1HNMR (400MHz, CDCl3, ppm) δ6.61(d,J=3.5Hz,1H),6.48(d,J=3.5Hz,1H),4.89–4.76(m,1H),4.37(t,J=5.7Hz,1H),4.37–4.27(m ,1H),4.04(q,J=7.2Hz,2H),2.21–2.06(m,4H),1.92–1.81(m,4H),1.79–1.63(m,6H),1.57–1.45(m,2H),1.27(t,J=7.2Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ159.06,149.56,147.94,117.56,101.77,100.54,55.38,53.84,35.20,33.35,32.60,24.37,23.86,13.10.

[0103] Example 4

[0104]

[0105] Preparation of 2-(cyclopentylamino)-3-ethyl-7-(4-methoxyphenyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 4):

[0106] The preparation process is the same as that in Example 1, except that 3-bromopropylene in step 5 is replaced with 4-bromomethoxybenzene to prepare 2-(cyclopentylamino)-3-ethyl-7-(4-methoxybenzene)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 4).

[0107] 1 HNMR (400MHz, CDCl3, ppm) δ7.20(d,J=7.7Hz,2H),6.83(d,J=7.5Hz,2H),6.51(d,J=2.7Hz,2H),6.48(d,J=3.0Hz,2H),5.10(s,2H),4.43( s,1H),4.39–4.30(m,1H),4.10–4.00(m,2H),3.77(s,3H),2.15–2.09(m,2H),1.76–1.65(m,4H),1.55–1.48(m,2H),1.28(t,J=7.4Hz,3H). 13CNMR (101MHz, CDCl3, ppm) δ159.00,150.04,148.17,130.01,129.04,119.81 ,113.94,102.34,100.20,55.22,53.86,47.22,35.25,33.32,23.83,13.06.

[0108] Example 5

[0109]

[0110] Preparation of 2-(cyclopentylamino)-3-ethyl-7-(pyridine-3-methylene)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 5):

[0111] The preparation process is the same as that in Example 1, except that 3-bromopropylene in step 5 is replaced with 3-bromomethylpyridine to prepare 2-(cyclopentylamino)-3-ethyl-7-(pyridine-3-methylene)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 5).

[0112] 1 HNMR (400MHz, CDCl3, ppm) δ8.59(s,1H),8.50(d,J=2.7Hz,1H),7.52(d,J=7.6Hz,1H),7.26–7.15(m,1H),6.56–6.44(m,2H),5.16(s,2 H),4.46(s,1H),4.36–4.23(m,1H),4.08–3.97(m,2H),2.11–2.03(m,2H),1.73–1.60(m,4H),1.52–1.42(m,2H),1.27(t,J=6.9Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ158.94,150.21,149.11,148.95,148.31,135.31,133 .46,123.55,119.63,102.91,100.35,53.86,45.47,35.31,33.30,23.82,13.00.

[0113] Example 6

[0114]

[0115] Preparation of 3-ethyl-7-isopropyl-2-(isopropylamino)-3,7-dihydro-4H-pyrrolo[2,3-d]pyridin-4-one (Compound 6):

[0116] The preparation process is the same as that in Example 2, except that the cyclopentylamine in step 6 is replaced by isopropylamine to prepare 3-ethyl-7-isopropyl-2-(isopropylamino)-3,7-dihydro-4H-pyrrolo[2,3-d]pyridin-4-one (Compound 6).

[0117] 1 HNMR(400MHz, CDCl3, ppm) δ6.62(d,J=3.5Hz,1H),6.48(d,J=3.5Hz,1H),4.80–4.66(m, 1H),4.30–4.17(m,2H),4.05(q,J=7.2Hz,2H),1.44(d,J=6.8Hz,6H),1.32–1.24(m,9H). 13 CNMR (101MHz, CDCl3, ppm) δ159.10,149.21,147.47,116.46,101.81,100.41,45.73,43.75,35.11,22.76,22.61,13.06.

[0118] Example 7

[0119]

[0120] Preparation of 2-(cyclopropylamino)-3-ethyl-7-isopropyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 7):

[0121] The preparation process is the same as that of Example 2, except that the cyclopentylamine in step 6 is replaced by cyclopropylamine to prepare 2-(cyclopropylamino)-3-ethyl-7-isopropyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 7).

[0122] 1 HNMR(400MHz, CDCl3)δ6.65(d,J=3.5Hz,1H),6.50(d,J=3.5Hz,1H),4.83–4.74(m,2H),4.05–3.97(m,2H ),2.89–2.79(m,1H),1.46(d,J=6.8Hz,6H),1.25(t,J=7.2Hz,3H),0.86–0.79(m,2H),0.60–0.53(m,2H). 13 CNMR(101MHz, CDCl3)δ159.12,150.69,147.43,116.90,101.70,100.89,45.97,35.39,24.82,22.52,13.15,7.16.

[0123] Example 8

[0124]

[0125] Preparation of 2-((4,4-difluorocyclohexane)amino)-3-ethyl-7-isopropyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 8):

[0126] The preparation process is the same as that of Example 2, except that the cyclopentylamine in step 6 is replaced with 4,4-difluorocyclohexylamine hydrochloride to prepare 2-((4,4-difluorocyclohexane)amino)-3-ethyl-7-isopropyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 8).

[0127] 1 HNMR (400MHz, CDCl3, ppm) δ6.64(d,J=3.5Hz,1H),6.50(d,J=3.5Hz,1H),4.76–4.65(m,1H),4.31(d,J=6.7Hz,1H),4.06 (q,J=7.2Hz,3H),2.24–2.11(m,4H),2.04–1.85(m,2H),1.73–1.62(m,2H),1.45(d,J=6.7Hz,6H),1.29(t,J=7.2Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ158.96,148.99,147.07,116.79,102.00,100. 79,48.67,45.86,35.26,32.48,32.24,31.99,28.60,28.51,22.68,13.14.

[0128] Example 9

[0129]

[0130] Preparation of 3-ethyl-7-isopropyl-2-(phenylamino)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 9):

[0131] Synthesis route:

[0132]

[0133] Compound 2-1 (1mmol, 1.0equiv.), aniline (1.3mmol, 1.3equiv.), methanesulfonic acid (2-dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium (II) (0.05mmol, 0.5equiv.), sodium tert-butoxide (1.5mmol, 1.5equiv.) were dissolved in 1,4-dioxane (5mL) and reacted at 80°C under nitrogen protection for 3 hours. After the reaction was completed, it was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product was purified by flash silica gel chromatography with ethyl acetate / petroleum ether = 1:2 (V / V) as the eluent to obtain a white solid (yield 61%).

[0134] 1 HNMR (400MHz, DMSO-d6, ppm) δ8.49(s,1H),7.60(d,J=7.7Hz,2H),7.33(t,J=7.9Hz,2H),7.06(t,J=7.4Hz,1H),6.92(d,J= 3.5Hz,1H),6.30(d,J=3.5Hz,1H),4.62–4.48(m,1H),4.25(q,J=7.0Hz,2H),1.35(d,J=6.7Hz,6H),1.20(t,J=7.0Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ159.04,147.05,146.27,139.00,128.74,123.43,121.05,121.03,117.44,102.17,102.05,45.97,35.96,22.69,13.55.

[0135] Example 10

[0136]

[0137] Preparation of 3-ethyl-7-isopropyl-2-(thiophene-3-methylene)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 10):

[0138] Synthesis route:

[0139]

[0140] Compound 2-1 (1mmol, 1.0equiv.), 3-aminothiophene hydrochloride (1.5mmol, 1.5equiv.), p-toluenesulfonic acid monohydrate (4mmol, 4.0equiv.) were dissolved in 1,4-dioxane (5mL) and reacted at 101°C for 12 hours. After the reaction was completed, it was diluted with water, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product was purified by rapid silica gel chromatography with ethyl acetate / petroleum ether = 1:2 (V / V) as the eluent to obtain a white solid (yield 25%).

[0141] 1 HNMR(400MHz,DMSO-d6,ppm)δ8.90(s,1H),7.73(dd,J=3.2,1.3Hz,1H),7.47(dd,J=5.2,3.2Hz,1H),7.40(dd,J=5.3,1.4Hz,1H),6 .95(d,J=3.5Hz,1H),6.33(d,J=3.5Hz,1H),4.79–4.65(m,1H),4.24(q,J=7.0Hz,2H),1.41(d,J=6.7Hz,6H),1.19(t,J=7.0Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ158.81,146.55,146.34,136.38,124.34,122.20,117.21,108.92,102.35,101.74,46.03,35.76,22.76,13.48.

[0142] Embodiment 11

[0143]

[0144] Preparation of 7-cyclopropyl-3-ethyl-2-(isopropylamino)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 11):

[0145] Synthesis route:

[0146]

[0147] Step 1: Synthesis of Intermediate 11-1

[0148] Compound 1-4 (1mmol, 1.0equiv.), cyclopropylboronic acid (2mmol, 2.0equiv.), copper acetate (0.5mmol, 0.5equiv.), lithium hydroxide (0.5mmol, 0.5equiv.) were dissolved in dichloromethane (5mL) and reacted overnight at room temperature. After the reaction was completed, it was diluted with water, extracted with dichloromethane, and the organic phase was washed with semi-saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product was purified by rapid silica gel chromatography with ethyl acetate / petroleum ether = 1:5 (V / V) as the eluent to obtain a white solid (yield 72%).

[0149] 1 HNMR(400MHz, CDCl3) δ6.77(d,J=3.5Hz,1H),6.56(d,J=3.4Hz,1H),4.44–4.20(m,2 H),3.49–3.33(m,1H),1.36(t,J=7.1Hz,3H),1.12–1.05(m,2H),1.03–0.95(m,2H). 13 CNMR (101MHz, CDCl3) δ158.47,146.43,143.10,123.42,106.82,102.38,41.61,26.83,13.82,6.40.

[0150] Step 2: Synthesis of compound 11

[0151] The preparation process is the same as step 6 of Example 6.

[0152] 1 HNMR(400MHz, CDCl3)δ6.51(d,J=3.5Hz,1H),6.42(d,J=3.5Hz,1H),4.35–4.20(m,2H),4.09–3 .99(m,2H),3.32–3.22(m,1H),1.30(d,J=6.2Hz,6H),1.27(t,J=7.2Hz,3H),0.99–0.90(m,4H). 13 CNMR(101MHz, CDCl3)δ159.04,149.64,149.38,120.01,101.66,100.55,43.87,35.13,26.25,22.72,13.00,6.01.

[0153] Example 12

[0154]

[0155] Preparation of 7-cyclopropyl-2-(cyclopropylamino)-3-ethyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 12):

[0156] The preparation process is the same as that of Example 11, except that the isopropylamine in step 2 is replaced by cyclopropylamine to prepare 7-cyclopropyl-2-(cyclopropylamino)-3-ethyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 12).

[0157] 1 HNMR (400MHz, CDCl3) δ6.54(d,J=3.5Hz,1H),6.43(d,J=3.5Hz,1H),4.89(s,1H),4.16–3.95(m,2H),3.39–3.29(m,1H) ,2.92–2.82(m,1H),1.24(t,J=7.2Hz,3H),1.06–1.02(m,2H),1.01–0.95(m,2H),0.87–0.77(m,2H),0.62–0.54(m,2H). 13 CNMR(101MHz, CDCl3)δ158.98,150.92,149.47,120.31,101.68,101.01,35.38,26.40,24.81,13.09,7.19,6.04.

[0158] Embodiment 13

[0159]

[0160] Preparation of 3-(3-ethyl-2-(isopropylamino)-4-oxo-3,4-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propanoic acid (Compound 13):

[0161] Synthesis route:

[0162]

[0163] Step 1: Synthesis of Intermediate 13-1

[0164] Compound 1-4 (1mmol, 1.0equiv.), ethyl 1-bromopropionate (1.2mmol, 1.2equiv.), tetrabutylammonium bromide (0.1mmol, 0.10equiv.) were dissolved in dichloromethane (5mL), and an equal volume of 50% sodium hydroxide solution was added dropwise, and stirred at room temperature for 4 hours. After the reaction was completed, it was diluted with water, extracted with dichloromethane, and the organic phase was washed with half-saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product was purified by rapid silica gel chromatography with ethyl acetate / petroleum ether = 1:5 (V / V) as the eluent to obtain a colorless oil (yield 30%).

[0165] 1 HNMR (400MHz, CDCl3, ppm) δ6.85 (d, J=3.4Hz, 1H), 6.59 (d, J=3.4Hz, 1H), 4.42–4.30 (m, 4H) ,4.13(q,J=7.1Hz,2H),2.81(t,J=6.7Hz,2H),1.36(t,J=7.1Hz,3H),1.22(t,J=7.2Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ170.88,158.42,144.88,143.17,123.91,106.51,102.84,60.94,41.61,40.67,35.20,14.09,13.81.

[0166] Step 2: Synthesis of Intermediate 13-2

[0167] The preparation process is the same as step 6 of Example 6.

[0168] 1 HNMR(400MHz, CDCl3, ppm) δ6.56(d,J=3.5Hz,1H),6.45(d,J=3.4Hz,1H),4.31–4.19(m,4H ),4.13(q,J=7.1Hz,2H),4.04(q,J=7.2Hz,2H),2.80(t,J=6.9Hz,2H),1.31–1.19(m,12H). 13 CNMR (101MHz, CDCl3, ppm) δ171.34,159.00,149.53,148.04,120.38,102.02,100.40,60.73,43.80,40.21,35.17,35.05,22.80,14.09,13.01.

[0169] Step 3: Synthesis of compound 13

[0170] Compound 10-2 (1 mmol, 1.0 equiv.) and potassium hydroxide (4 mmol, 4.0 equiv.) were dissolved in ethyl acetate (5 mL) and stirred at room temperature for 2 hours. TLC monitored the completion of the reaction, and 1 mol / L hydrochloric acid solution was added dropwise to the reaction mixture to adjust the pH to 5. A pink solid was precipitated, which was filtered, washed, and dried to obtain a portion of the target compound. The filtrate was extracted with dichloromethane, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a pink solid (yield 90%).

[0171] 1 HNMR(400MHz,DMSO-d6,ppm)δ12.35(s,1H),6.70(d,J=3.4Hz,1H),6.37(d,J=7.4Hz,1H),6.18(d,J=3.4Hz,1H),4.26–4.1 7(m,1H),4.13(t,J=7.0Hz,2H),4.02(q,J=7.0Hz,2H),2.72(t,J=7.0Hz,2H),1.23(d,J=6.5Hz,6H),1.09(t,J=7.0Hz,3H). 13 CNMR(101MHz,DMSO-d6,ppm)δ172.38,158.03,149.80,147.83,120.39,100.94,99.00,43.22,34.41,34.23,28.99,22.05,13.11.

[0172] Embodiment 14

[0173]

[0174] Preparation of 4-(3-ethyl-2-(isopropylamino)-4-oxo-3,4-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)butanoic acid (Compound 14):

[0175] The preparation process is the same as that of Example 13, except that 1-bromopropionic acid ethyl ester in step 1 is replaced with 1-bromobutyric acid ethyl ester to prepare 4-(3-ethyl-2-(isopropylamino)-4-oxo-3,4-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)butyric acid (Compound 14).

[0176] 1HNMR(400MHz,DMSO-d6,ppm)δ12.07(s,1H),6.70(d,J=3.4Hz,1H),6.34(d,J=7.4Hz,1H),6.21(d,J=3.4Hz,1H),4.28–4.15(m,1H), 4.02(q,J=7.0Hz,2H),3.95(t,J=6.7Hz,2H),2.15(t,J=7.4Hz,2H),2.01–1.87(m,2H),1.22(d,J=6.5Hz,6H),1.09(t,J=6.9Hz,3H). 13 CNMR(101MHz,DMSO-d6,ppm)δ173.84,158.08,149.80,148.04,120.25,101.12,98.89,43.18,42.80,34.25,30.74,25.34,22.11,13.15.

[0177] Embodiment 15

[0178]

[0179] Preparation of tert-butyl 4-(2-(cyclopentylamino)-3-ethyl-4-oxo-3,4-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate (Compound 15):

[0180] Synthesis route:

[0181]

[0182] Step 1: Synthesis of Intermediate 15-1

[0183] 2,4-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (21mmol, 1.0equiv.), triphenylphosphine (27.3mmol, 1.3equiv.), N-Boc-4-hydroxypiperidine (27.3mmol, 1.3equiv.) were dissolved in tetrahydrofuran (25mL) and stirred at room temperature under nitrogen for 30 minutes. After cooling to 0°C, di-tert-butyl azodicarboxylate (27.3mmol, 1.3equiv.) was added dropwise, stirred for 30 minutes, and then returned to room temperature. After stirring at room temperature for 5 hours, the mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product was purified by flash silica gel chromatography with ethyl acetate / petroleum ether = 1:5 (V / V) as the eluent to obtain a white solid (yield 60%).

[0184] 1HNMR(400MHz, CDCl3, ppm) δ7.28(d,J=3.7Hz,1H),6.63(d,J=3.7Hz,1H),4.85(ddt,J=12.1,8.0,4.2H z,1H),4.39–4.25(m,2H),2.94(t,J=25.0Hz,2H),2.05–1.99(m,2H),1.92–1.84(m,2H),1.49(s,9H). 13 CNMR (101MHz, CDCl3, ppm) δ154.52,152.70,151.70,151.43,126.46,116.47,100.47,80.13,52.38,32.20,28.40,28.13.

[0185] Step 2: Synthesis of Intermediate 15-2

[0186] The preparation process is the same as step 2 of Example 1.

[0187] 1 HNMR (400MHz, CDCl3, ppm) δ12.74 (s, 1H), 6.91 (d, J = 3.6Hz, 1H), 6.74 (d, J = 3.5Hz, 1H), 4.68 (tt, J = 12. 0,4.1Hz,1H),4.30(s,2H),2.90(t,J=15.4Hz,2H),2.05–1.94(m,2H),1.93–1.79(m,2H),1.48(s,9H). 13 CNMR (101MHz, CDCl3, ppm) δ161.74,154.58,146.80,141.32,120.41,106.29,103.29,80.02,52.27,43.51,32.43,28.40.

[0188] Step 3: Synthesis of Intermediate 15-3

[0189] The preparation process is the same as step 3 of Example 1.

[0190] 1 HNMR (400MHz, CDCl3, ppm) δ6.84(d,J=3.5Hz,1H),6.65(d,J=3.4Hz,1H),4.68–4.55(m,1H),4.38–4.18( m,4H),2.90(t,J=13.0Hz,2H),2.01–1.94(m,2H),1.91–1.76(m,2H),1.48(s,9H),1.36(t,J=7.1Hz,3H). 13CNMR (101MHz, CDCl3, ppm) δ158.41,154.56,144.41,143.05,119.97,106.33,103.26,79.95,51.90,43.11,41.61,32.47,28.39,13.83.

[0191] Step 4: Synthesis of compound 15

[0192] The preparation process is the same as step 6 of Example 1.

[0193] 1 HNMR(400MHz, CDCl3, ppm) δ6.58(d,J=3.5Hz,1H),6.49(d,J=3.5Hz,1H),4.39–4.36(m,1H),4.34–4.25(m,2H),4.04(q,J=7.2Hz, 2H),2.86(s,2H),2.16–2.05(m,2H),2.02–1.86(m,4H),1.79–1.59(m,6H),1.58–1.47(m,2H),1.48(s,9H),1.28(t,J=7.3Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ159.00,153.81,149.71,147.62,117.01,102.18,1 00.66,79.76,53.88,52.32,43.20,35.27,33.41,31.95,28.41,23.98,13.08.

[0194] Example 16

[0195]

[0196] Preparation of 7-(1-allylpiperidin-4-yl)-2-(cyclopentylamino)-3-ethyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 16):

[0197] Synthesis route:

[0198]

[0199] Step 1: Synthesis of Intermediate 16-1

[0200] The preparation process is the same as step 4 of Example 1.

[0201] 1HNMR (400MHz, DMSO-d6, ppm) δ12.02 (s, 1H), 7.09 (d, J = 3.4Hz, 1H), 6.49 (d, J = 3.3Hz, 1H), 4.23 (q, J = 7.1Hz, 2H), 1.26 (t, J = 7.0Hz, 3H). 13 CNMR (101MHz, CDCl3, ppm) δ159.09,145.34,143.69,121.17,106.32,103.95,42.33,13.70.

[0202] Step 2: Synthesis of Intermediate 16-2

[0203] The preparation process is the same as step 5 of Example 1.

[0204] 1 HNMR(400MHz, CDCl3, ppm) δ6.91(d,J=3.5Hz,1H),6.64(d,J=3.5Hz,1H),5.97–5.82(m,1H),5.28–5.11(m,2H),4.5 7–4.44(m,1H),4.40–4.30(m,2H),3.12–3.04(m,4H),2.24–2.13(m,2H),2.07–1.94(m,4H),1.36(t,J=7.1Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ158.49,144.49,142.91,134.72,120.18,118.36,106.25,103.11,61.56,52.61,51.68,41.58,32.61,13.86.

[0205] Step 3: Synthesis of compound 16

[0206] The preparation process is the same as step 6 of Example 1.

[0207] 1 HNMR (400MHz, CDCl3, ppm) δ6.66(d,J=3.6Hz,1H),6.50(d,J=3.5Hz,1H),6.02–5.87(m,1H),5.29–5.18(m,2H),4.43–4.28(m,3H),4.04(q,J=7.2 Hz,2H),3.16–3.10(m,4H),2.23(t,J=11.6Hz,2H),2.16–2.06(m,4H),2 .04(s,2H),1.77–1.66(m,4H),1.56–1.49(m,2H),1.28(t,J=7.9Hz,3H).13 CNMR (101MHz, CDCl3, ppm) δ159.00,149.78,147.68,118.93,118.92,116.80,1 02.19,100.50,61.41,53.87,52.78,51.22,35.27,33.45,31.81,23.99,13.10.

[0208] Embodiment 17

[0209]

[0210] Preparation of 7-(1-benzylpiperidin-4-yl)-2-(cyclopentylamino)-3-ethyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 17):

[0211] The preparation process is the same as that of Example 16, except that allyl bromide in step 2 is replaced by benzyl bromide to prepare 7-(1-benzylpiperidin-4-yl)-2-(cyclopentylamino)-3-ethyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 17).

[0212] 1 HNMR(400MHz, CDCl3, ppm) δ7.41–7.26(m,5H),6.65(d,J=3.5Hz,1H),6.49(d,J=3.5Hz,1H),4.41–4.25(m,3H),4.04(q,J=7.1Hz,2H),3.61(s,2H ),3.06(d,J=11.1Hz,2H),2.27–2.18(m,2H),2.18–2.06(m,4H),2.03–1 .94(m,2H),1.80–1.62(m,4H),1.58–1.47(m,2H),1.28(t,J=7.4Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ159.02,149.68,147.64,129.22,128.33,127.27,117.0 2,102.00,100.53,62.81,53.86,52.90,51.57,35.24,33.43,31.95,23.98,13.09.

[0213] Embodiment 18

[0214]

[0215] Preparation of 7-(1-acetylpiperidin-4-yl)-2-(cyclopentylamino)-3-ethyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 18):

[0216] Synthesis route:

[0217]

[0218] Step 1: Synthesis of Intermediate 18-1

[0219] Compound 13-1 (1mmol, 1.0equiv.) and diisopropylethylamine (1.5mol, 1.5equiv.) were dissolved in dichloromethane, and acetyl chloride (1.5mol, 1.5equiv.) was added dropwise at 0°C and stirred overnight. After the reaction was completed, the mixture was diluted with water, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product was purified by flash silica gel chromatography with ethyl acetate / petroleum ether = 3:1 (V / V) as the eluent to obtain a white solid (yield 70%).

[0220] 1 HNMR (400MHz, DMSO-d6, ppm) δ7.31(d,J=3.0Hz,1H),6.52(d,J=3.4Hz,1H),4.68–4.61(m,1H),4.54(d,J=13.4Hz,1H),4.21(q,J=21.8 Hz,2H),3.94(d,J=13.9Hz,1H),3.24(t,J=12.9Hz,1H),2.70(t,J=12.6Hz,1H),2.04(s,3H),1.97–1.73(m,4H),1.23(t,J=3.6Hz,3H). 13 CNMR(101MHz,DMSO-d6,ppm)δ168.10,157.43,143.75,142.58,121.80,105.45,102.34,51.57,45.53,44.91,41.04,32.31,31.64,21.30,13.62.

[0221] Step 2: Synthesis of compound 18

[0222] The preparation process is the same as step 6 of Example 1.

[0223] 1HNMR (400MHz, CDCl3, ppm) δ6.54 (d, J=3.5Hz, 1H), 6.47 (d, J=3.0Hz, 1H), 4.85–4. 76(m,1H),4.53–4.40(m,2H),4.34–4.21(m,1H),4.03(q,J=7.2Hz,2H),4.00–3.9 1(m,1H),3.22(td,J=13.1,2.8Hz,1H),2.67(td,J=12.8,3.6Hz,1H),2.13(s,3H) ,2.10–1.88(m,6H),1.78–1.63(m,4H),1.58–1.47(m,2H),1.26(t,J=8.1Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ168.83,158.91,149.72,147.58,116.99,102.34,100.73,53. 91,52.28,45.85,41.11,35.26,33.39,33.35,32.48,31.44,29.64,23.89,21.40,13.03.

[0224] Embodiment 19

[0225]

[0226] Preparation of 2-(cyclopentylamino)-3-ethyl-7-(1-propionylpiperidin-4-yl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 19):

[0227] The preparation process is the same as that of Example 18, except that the acetyl chloride in step 1 is replaced by propionyl chloride to prepare 2-(cyclopentylamino)-3-ethyl-7-(1-propionylpiperidin-4-yl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 19).

[0228] 1HNMR(400MHz,DMSO-d6,ppm)δ6.78(d,J=3.5Hz,1H),6.42(d,J=6.2Hz,1H),6.19(d ,J=3.4Hz,1H),4.55(d,J=13.2Hz,1H),4.47–4.35(m,1H),4.26–4.14(m,1H),4.08 –3.94(m,3H),3.11(t,J=13.1Hz,1H),2.61(t,J=12.6Hz,1H),2.42–2.27(m,2H),2 .05–1.80(m,6H),1.72–1.46(m,6H),1.07(t,J=6.9Hz,3H),1.00(t,J=7.4Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ172.13,158.93,149.71,147.62,117.03,102.26,100.69 ,53.87,52.40,44.87,41.26,35.24,33.34,32.53,31.50,26.50,23.91,13.03,9.52.

[0229] Embodiment 20

[0230]

[0231] Preparation of 1-(3-(3-ethyl-2-(isopropylamino)-4-oxo-3,4-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)azetidine-1-carbonyl)cyclopropane-1-carbonitrile (Compound 20):

[0232] Synthesis route:

[0233]

[0234] Step 1: Synthesis of Intermediate 20-1

[0235] The preparation process is the same as step 1 of Example 15, except that N-Boc-4-hydroxypiperidine in step 1 is replaced by N-Boc-3-hydroxyazetidine to prepare intermediate 20-1.

[0236] 1 HNMR (400MHz, CDCl3, ppm) δ7.56 (d, J = 3.7Hz, 1H), 6.71 (d, J = 3.7Hz, 1H), 5.5 7(tt,J=8.0,5.0Hz,1H),4.55–4.46(m,2H),4.23–4.14(m,2H),1.48(s,9H). 13CNMR (101MHz, CDCl3, ppm) δ156.08,152.99,152.19,152.18,126.32,116.52,101.62,80.60,56.64,43.54,28.29.

[0237] Step 2: Synthesis of Intermediate 20-2

[0238] The preparation process is the same as step 2 of Example 1 1 HNMR(400MHz,DMSO-d6,ppm)δ7.46(d,J=3.6Hz,1H),6.56(d,J=3.5Hz,1H),5.3 6(tt,J=8.2,5.6Hz,1H),4.33(t,J=8.6Hz,2H),4.18–4.14(m,2H),1.42(s,9H). 13 CNMR(101MHz,DMSO-d6,ppm)δ158.82,155.55,146.71,141.93,122.03,106.47,102.66,79.00,56.07,43.50,28.06.

[0239] Step 3: Synthesis of intermediate 20-3

[0240] The preparation process is the same as step 3 of Example 1 1 HNMR (400MHz, CDCl3, ppm) δ7.10(d,J=3.6Hz,1H),6.71(d,J=3.5Hz,1H),5.37(tt,J=8.2,5.2Hz,1H ),4.45–4.39(m,2H),4.33(q,J=7.1Hz,2H),4.18–4.13(m,2H),1.46(s,9H),1.34(t,J=7.1Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ158.26,156.08,145.36,143.63,119.93,106.57,104.39,80.27,56.86,43.15,41.66,28.28,13.75.

[0241] Step 4: Synthesis of Intermediate 20-4

[0242] The preparation process is the same as step 4 of Example 1

[0243] Step 5: Synthesis of Intermediate 20-5

[0244] Compound 20-4 (1mmol, 1.0equiv.), 1-cyano-1-cyclopropanecarboxylic acid (2mmol, 2.0equiv.) were dissolved in dichloromethane, and diisopropylethylamine (1.5mmol, 1.5equiv.), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.5mmol, 1.5equiv.), and 1-hydroxybenzotriazole (1.5mmol, 1.5equiv.) were added dropwise at room temperature. After stirring at room temperature for 3 hours, the mixture was diluted with water, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product. The crude product was purified by flash silica gel chromatography with ethyl acetate / petroleum ether = 1:2 (V / V) as the eluent to obtain a white solid (yield 50%).

[0245] 1 HNMR (400MHz, CDCl3, ppm) δ7.04(d,J=3.6Hz,1H),6.73(d,J=3.5Hz,1H),5.43(tt,J=8.4,5.5Hz,1H),5.12(s,1H),4.91(d,J=8.0H z,1H),4.55(d,J=8.3Hz,1H),4.41(s,1H),4.34(q,J=7.1Hz,2H),1.76–1.72(m,2H),1.53(d,J=4.0Hz,2H),1.35(t,J=7.1Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ165.64,158.19,145.38,143.82,120.34,119.73,107.02,104.51,60.33,56.77,43.95,41.74,18.33,13.72,11.09.

[0246] Step 6: Synthesis of compound 20

[0247] The preparation process is the same as step 6 of Example 6.

[0248] 1HNMR (400MHz, DMSO-d6, ppm) δ6.84(d,J=3.5Hz,1H),6.45(d,J=7.4Hz,1H),6.22(d,J=3.5Hz,1H),5.20(tt,J=8.4,6.0Hz,1H),4.92(d,J= 9.9Hz,2H),4.41–4.31(m,2H),4.21–4.06(m,1H),4.00(q,J=7.0Hz,2H),1.63–1.41(m,4H),1.18(d,J=12.8Hz,6H),1.06(t,J=7.0Hz,3H). 13 CNMR(101MHz,DMSO-d6,ppm)δ164.32,158.03,149.76,148.38,119.91,119.76,101 .65,99.88,59.12,55.92,44.40,43.35,34.32,22.12,17.41,16.89,13.05,10.71.

[0249] Embodiment 21

[0250]

[0251] Preparation of 3-ethyl-2-(isopropylamino)-7-(1-(6-methylpyridine)azetidin-3-yl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 21):

[0252] The preparation process is the same as that of Example 20, except that the 1-cyano-1-cyclopropanecarboxylic acid in step 5 is replaced with 6-methyl-2-pyridinecarboxylic acid to prepare 3-ethyl-2-(isopropylamino)-7-(1-(6-methylpyridine)azetidine-3-yl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one (Compound 21).

[0253] 1HNMR (400MHz, CDCl3, ppm) δ7.93 (d, J = 7.7Hz, 1H), 7.69 (t, J = 7.7Hz, 1H), 7.21 (d ,J=7.7Hz,1H),6.72(d,J=3.5Hz,1H),6.53(d,J=3.5Hz,1H),5.32–5.24(m,1H), 5.24–5.13(m,2H),4.66(d,J=6.9Hz,2H),4.35(d,J=7.1Hz,1H),4.20–4.10(m,1 H),4.03(q,J=7.2Hz,2H),2.51(s,3H),1.30–1.19(m,6H),1.11(d,J=6.5Hz,3H). 13 CNMR (101MHz, CDCl3, ppm) δ165.15,158.99,157.03,151.02,149.53,148.61,136.99,124.96,1 20.71,118.00,103.08,100.92,61.97,55.92,45.29,43.83,35.19,24.16,22.79,22.67,12.93.

[0254] Example 22 Phosphodiesterase VII (PDE7) enzyme inhibition activity test:

[0255] Using BRL-50481 as a control drug, the time-resolved fluorescence resonance energy transfer method was used to test the inhibitory activity of the compound on PDE7A. The compound was dissolved in DMSO to prepare a 10mM stock solution, which was then diluted with DMSO in a gradient manner and added to the final reaction system to ensure that the concentration of DMSO should be lower than 1%. The enzyme reaction system of PDE7A consists of PDE7A enzyme, BSA (bovine serum albumin), PDE7A-specific fluorescent substrate FAM-cAMP, reaction buffer, etc. When all the components are mixed together, react at room temperature for 60 minutes. After the reaction is completed, a specific phosphate-binding antibody is added and incubated at room temperature for another 60 minutes. The fluorescence polarization signal is then detected on MD's SpectraMaxM5 multifunctional microplate reader, with an excitation wavelength of 485nm and an emission wavelength of 528nm. Substitute the values ​​into the following formula to calculate the percentage of activity:

[0256] % activity = {(FP drug – FP background) / (FP enzyme – FP background)} × 100%

[0257] The IC50 value of the compound was tested, and the inhibition rate of the compound at different concentrations on the enzyme was fitted with a dose-effect curve by nonlinear regression and normalization using PrismGraphPad software. The inhibitory activity of the compound on PDE7A is shown in Table 1 below.

[0258] Table 1 Inhibitory activity of compounds against PDE7A (IC50, nM) a

[0259]

[0260]

[0261] a All data are the average of three experiments.

[0262] b Inhibitory activity of compounds against PDE7A at 1000 nM concentration. All data are the average of three experiments.

[0263] As shown in Table 1, the inhibitory activity of compounds 1, 2, 4, 5, 6, 9, 15, 16, 17, 18, and 19 on PDE7A is greater than that of the positive drug BLR-50481. The anti-PDE7A activity of compounds 11, 12, 13, and 21 is comparable to that of the positive drug BLR-50481.

[0264] Example 23 Anti-alcoholic hepatitis effect of compound 6

[0265] Alcoholic hepatitis is a special category of alcoholic liver disease characterized by globular hepatocytes containing amorphous eosinophilic inclusions (called Mallory bodies) surrounded by neutrophils. The presence of large fat globules in the hepatocytes (also called steatosis) is also common in alcoholic hepatitis.

[0266] The improvement effect of PDE7 inhibitor compound 6 on alcoholic hepatitis was preliminarily confirmed. Female C57 mice aged 8-10 weeks were used for a 16-day alcoholic hepatitis model test. Five days before the experiment, all mice were fed with ordinary liquid feed, and the feed was supplemented every afternoon to adapt the mice to this feeding method. On the afternoon of the 6th day of the experiment, the mice were randomly divided into 3 groups, namely the control group, the alcohol group and the drug group (10 mg / kg). From the 6th to the 15th day of the experiment, the blank group was fed with ordinary liquid feed. From the 6th to the 8th day of the experiment, the other two groups were fed with alcohol liquid feed containing 1% alcohol; from the 9th to the 11th day of the experiment, the other two groups were fed with alcohol liquid feed containing 2% alcohol; from the 12th to the 15th day of the experiment, the other two groups were fed with alcohol liquid feed containing 4% alcohol. The experimental drug was prepared into a certain concentration of liquid medicine, and the drug group was gavaged according to the dose before supplementing the feed in the afternoon of the 11th to the 15th day of the experiment, and the drug was administered for a total of 5 days. At 9:00 on the 16th day of the experiment, the blank group was gavaged with 45% maltodextrin solution at a dose of 9 g / kg, and the other four groups were gavaged with 31.5% alcohol solution at a dose of 5 g / kg; at 18:00 on the same day, the mice were anesthetized, and their eyes were removed for blood and liver extraction.

[0267] During the experiment, each mouse was weighed and monitored every afternoon when the feed was replenished. Figure 1 As shown. The whole blood obtained by removing the mouse eyeball was placed in a centrifuge tube containing sodium heparin. After being placed at room temperature for 1 hour, it was centrifuged at 4°C and 3000rpm for 10 minutes, and the supernatant was aspirated as plasma. The ALT, AST and TG levels of mouse plasma were tested using the alanine aminotransferase (GPT / ALT) test kit, aspartate aminotransferase (GOT / AST) test kit and triglyceride (TG) assay kit of Nanjing Jiancheng Haihao Biotechnology Co., Ltd. The results are shown in Figures 2 to 4 shown.

[0268] Three small pieces of mouse liver were cut from each liver with scissors and stored at -80°C for use in real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). Total RNA from mouse liver was extracted using Trizol reagent (Invitrogen; Thermo Fisher Scientific Inc.). Before further analysis, the quality of RNA was verified using an Eppendorf ultra-micro nucleic acid spectrometer. TM According to the instruction manual of RT Reagent kit (TakaraBio, Inc.), 1000 ng of total RNA and 20 μL of reaction volume were used for reverse transcription. The primers used in this experiment (Qingdao Biotechnology, China) IL-1β forward sequence, IL-1β reverse sequence, IL-6 forward sequence, IL-6 reverse sequence, TNF-α forward sequence, TNF-α reverse sequence, β-actin forward sequence, β-actin reverse sequence are shown in SEQ ID: No. 1 to SEQ ID: No. 8.

[0269] RT-qPCR was performed using the FastReal qPCR premix (SYBR Green) kit (Zhenzhi Bio, China) and Roche LightCycler 480. The thermal cycling conditions were: 95°C for 2 min, 95°C for 5 s, 60°C for 15 s, and 40 cycles. β-actin was used as the reference gene in this experiment. The experimental results are shown in Figures 5 to 7 The remaining liver was immersed in 4% paraformaldehyde and allowed to stand for 18 hours before being sent for sectioning and hematoxylin-eosin staining. Figure 8 The experimental results show that compound 6 can effectively improve the liver damage caused by alcohol.

[0270] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A pyrrolo[2,3-d]pyrimidine compound or a pharmaceutically acceptable salt thereof, characterized in that: Its structural formula is shown in the following formula (I): Wherein, R1 is selected from C 2-8 Alkenyl, C 1-8 Alkyl, C 3-8 Cycloalkyl, aryl C 1-8 Alkyl, heteroaryl C 1-8 Alkyl, C 1-8 Alkyl carboxyl, C 3-8 Heterocyclic group; wherein the aryl C 1-8 Alkyl, heteroaryl C 1-8 Alkyl, C 3-8 The heterocyclic group is unsubstituted or substituted with one or more selected from C 1-8 Alkoxy, C 1-6 Alkyl ester group, C 2-8 Alkenyl, aryl C 1-8 Alkyl, C 1-8 Alkylcarbonyl, C 3-8 The substituents of cycloalkylcarbonyl and heteroarylcarbonyl are substituted; wherein C 1-8 Alkylcarbonyl, C 3-8 Cycloalkylcarbonyl and heteroarylcarbonyl are unsubstituted or substituted with one or more selected from C 1-8 Alkyl or cyano substituents; R2 is selected from NR4R5; wherein R4 and R5 are each independently selected from hydrogen, C 3-8 Cycloalkyl, C 1-8 Alkyl, aryl, heteroaryl; wherein the C 3-8 Cycloalkyl, C 1-8 Alkyl, aryl, heteroaryl are unsubstituted or substituted with one or more substituents independently selected from halogen; R3 is selected from C 1-8 alkyl.

2. The pyrrolo[2,3-d]pyrimidine compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R1 is selected from C 2-6 Alkenyl, C 1-6 Alkyl, C 3-6 Cycloalkyl, aryl C 1-8 Alkyl, heteroaryl C 1-8 Alkyl, C 1-8 Alkyl carboxyl, C 3-8 Heterocyclic group; wherein the aryl C 1-8 The aryl group in the alkyl group is a 5-6 membered aromatic ring, and the heteroaryl C 1-8 The heteroaryl group in the alkyl group is a 5-6 membered aromatic heterocyclic ring; Wherein, the 5-6 membered aromatic ring, 5-6 membered aromatic heterocyclic ring, C 3-8 The heterocyclic group is unsubstituted or substituted with one or more selected from C 1-5 Alkoxy, C 1-6 Alkyl ester group, C 2-8 Alkenyl, benzyl, C 1-5 Alkylcarbonyl, C 3-6 Substituted by cycloalkylcarbonyl, pyridinecarbonyl; wherein C 1-5 Alkylcarbonyl, C 3-6 Cycloalkylcarbonyl and pyridinecarbonyl are unsubstituted or substituted with one or more selected from C 1-6 Alkyl or cyano substituents; R2 is selected from NR4R5; wherein R4 and R5 are each independently selected from hydrogen, C 3-8 Cycloalkyl, C 1-5 alkyl, 5-6 membered aromatic ring, 5-6 membered aromatic heterocyclic ring; wherein the C 3-8 Cycloalkyl, C 1-5 Alkyl, 5-6 membered aromatic ring, 5-6 membered aromatic heterocyclic ring are unsubstituted or substituted with one or more substituents independently selected from halogen; R3 is selected from C 1-5 alkyl.

3. The pyrrolo[2,3-d]pyrimidine compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R1 is selected from ethylene, propylene, butene, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl C 1-3 Alkyl, heteroaryl C 1-3 Alkyl, C 1-4 alkylcarboxyl, aminopiperidinyl, azetidinyl; wherein the heteroaryl C 1-3 The heteroaryl in the alkyl group is pyridyl, furyl, thienyl, pyrrolyl, thiazolyl, imidazolyl, pyrazinyl, pyrimidinyl; The phenyl, pyridyl, furyl, thienyl, pyrrolyl, thiazolyl, imidazolyl, pyrazinyl, pyrimidinyl, aminopiperidinyl, azetidinyl are unsubstituted or substituted with one or more substituents selected from methoxy, ethoxy, propoxy, tert-butyl ester, ethylene, propylene, butylene, benzyl, methylcarbonyl, ethylcarbonyl, propylcarbonyl, cyclopropylcarbonyl, pyridylcarbonyl; wherein cyclopropylcarbonyl and pyridylcarbonyl are unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl or cyano; R2 is selected from NR4R5; wherein R4 and R5 are each independently selected from hydrogen, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, phenyl, pyridyl, furyl, thienyl, pyrrolyl, thiazolyl, imidazolyl, pyrazinyl, pyrimidinyl; wherein the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, pyridyl, furyl, thienyl, pyrrolyl, thiazolyl, imidazolyl, pyrazinyl, pyrimidinyl are unsubstituted or substituted with one or more substituents independently selected from halogen; R3 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl.

4. The pyrrolo[2,3-d]pyrimidine compound or a pharmaceutically acceptable salt thereof according to claim 3, characterized in that: R1 is selected from R2 is selected from hydrogen, 5. The pyrrolo[2,3-d]pyrimidine compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: Select from any of the following structures:

6. A pharmaceutical composition, characterized in that The invention comprises the pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5; and a pharmaceutically acceptable excipient.

7. Use of the pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 or the pharmaceutical composition according to claim 6 in the preparation of a phosphodiesterase 7 inhibitor.

8. Use of the pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 or the pharmaceutical composition according to claim 6 in the preparation of a drug for preventing and treating phosphodiesterase 7-related diseases.

9. The use according to claim 8, characterized in that: The related diseases include one or more of inflammatory diseases, autoimmune diseases, addictive diseases, primary impulse control disorders, and neurodegenerative diseases.

10. Use of the pyrrole [2,3-d] pyrimidine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 or the pharmaceutical composition according to claim 6 in the preparation of a medicament for treating alcoholic hepatitis.

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