Tricyclic fused heterocycle PDE3 / 4 dual inhibitor as well as preparation method and application thereof

Abstract

The invention provides a tricyclic fused heterocyclic PDE3 / 4 dual inhibitor as well as a preparation method and application thereof, and particularly provides a compound with a structural formula I or a pharmaceutically acceptable form thereof, and each substituent group is defined in the specification. The compound or the pharmaceutically acceptable form of the compound can be used as a phosphodiesterase inhibitor and has relatively high activity. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a tricyclic fused heterocyclic phosphodiesterase inhibitor (especially a PDE3 / 4 dual inhibitor), and its preparation method and application. Background Art

[0002] Phosphodiesterase (PDE) belongs to the superfamily of enzymes, and contains at least 11 families and 22 subtypes, which are involved in intracellular and extracellular information transmission and functional regulation. PDE can catalyze the hydrolysis and ring opening of the second messengers cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) in cells to generate AMP and GMP respectively.

[0003] The PDE3 family is composed of two genes, namely PDE3A and PDE3B. The activity of PDE3 is mainly concentrated in alveolar macrophages, endothelial cells, and platelets in the respiratory system. PDE3 is involved in the regulation of various physiological activities in the body, such as relaxing vascular smooth muscle, antiplatelet aggregation, antithrombosis, cardiotonic, and anti-cell proliferation. Excessive use of PDE3 inhibitors will cause adverse reactions such as hypotension and tachycardia, which greatly limits their clinical application.

[0004] PDE4 is an enzyme that specifically hydrolyzes cAMP. Its family is composed of four subtypes: PDE4A, PDE4B, PDE4C, and PDE4D. Each subtype has a corresponding gene encoding and has different cell distributions and functions. PDE4 is mainly distributed in airway smooth muscle cells, inflammatory cells, and immune cells to regulate the intracellular cAMP level. Most of the current clinically used PDE4 inhibitors have certain side effects, such as gastrointestinal reactions such as nausea and vomiting, and even depression.

[0005] In view of the limitations of using PDE3 or PDE4 inhibitors alone and the side effects of PDE inhibitors, inhaled dual inhibition of PDE3 / 4 seems to be a more attractive way to target and treat the key pathological features of COPD and asthma. Currently, there is evidence showing that inhaled dual-target PDE3 / 4 inhibitors have a synergistic inhibitory effect, including synergistic anti-inflammatory and bronchodilation effects.

[0006] CN100415743C discloses a pyrimido[6,1-a]isoquinolin-4-one derivative:

[0007]

[0008] This general formula compound is used as a PDE inhibitor for treating respiratory diseases such as asthma, has a longer duration of action compared with trequinsin, and does not have the very bitter taste of trequinsin.

[0009] CN112368281A discloses a class of tricyclic compounds as dual PDE3 / PDE4 inhibitors

[0010]

[0011] The compounds of the general formula can be used to prepare drugs related to PDE3 / PDE4, especially drugs for chronic obstructive pulmonary disease (COPD).

[0012] There is still an urgent need in the art for new PDE3 / PDE4 inhibitors, especially PDE3 / PDE4 inhibitors with high activity and good drug-likeness. Summary of the Invention

[0013] One object of the present invention is to provide a new compound as a PDE inhibitor.

[0014] Another object of the present invention is to provide a preparation method of the compound.

[0015] Another object of the present invention is to provide an application of the compound.

[0016] Another object of the present invention is to provide a pharmaceutical composition containing the compound and its application.

[0017] Another object of the present invention is to provide an intermediate for preparing the compound.

[0018] Another object of the present invention is to provide a preparation method of the intermediate.

[0019] <First aspect>

[0020] The present invention provides a compound of formula I or a pharmaceutically acceptable form thereof,

[0021]

[0022] Wherein:

[0023] R 1 、R 2 Each independently selected from H, C 1-6 linear alkyl, C 3-6 branched alkyl and C 3-6 cycloalkyl; the linear alkyl, branched alkyl or cycloalkyl is optionally further substituted by 0 to 4 substituents selected from D, F, Cl, Br, I, OH, =O, NH 2 、CN、COOH、C 1-4 alkyl, C 1-4 alkoxy;

[0024] R 3 、R 4 、R5 Each independently selected from H, halogen, CN, C 1-6 alkoxy, C 1-6 linear alkyl, C 3-6 branched alkyl, and C 3-6 cycloalkyl; the alkoxy, linear alkyl, branched alkyl or cycloalkyl is optionally further substituted by 0 to 4 substituents selected from D, F, Cl, Br, I, OH, ═O, NH 2 , CN, COOH, C 1-4 alkyl, C 1-4 alkoxy;

[0025] L is selected from and, n is 0, 1 or 2; k is 0, 1, 2 or 3; the H in the L is optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, OH, ═O, NH 2 , CN, COOH, C 1-4 alkyl, C 1-4 alkoxy; wherein, R 9 , R 10 each independently selected from H, C 1-6 linear alkyl, C 3-6 branched alkyl, C 3-6 cycloalkyl, C 3-6 heterocycloalkyl, C 6-10 aryl, C 5-10 heteroaryl and COOCH 3 , and R 9 , R 10 are not simultaneously H; the heteroaryl contains 1-3 heteroatoms selected from N, O and S;

[0026] R 6 is selected from wherein, R 11 is selected from amino, C 1-6 alkoxy, C 3-12 cycloalkyl, C 6-10 aryl, C 5-10 heterocyclic group, the heterocyclic group contains 1-3 heteroatoms selected from N, O and S, the amino, alkoxy, cycloalkyl, aryl, heteroaryl is optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, OH, ═O, NH 2 , CN, COOH, CF 3 , C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl;

[0027] Optionally, R 6 forms

[0028] R 7 、R 8 are each independently selected from H, ═O, halogen, NH 2 , CN, C 1-6 linear alkyl, C 3-6 branched alkyl, C 3-6 cycloalkyl and

[0029] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 1 , R 2 are each independently selected from CH 3 , CHF 2 , CD 3 or C 3-6 cycloalkyl.

[0030] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 1 , R 2 are respectively CD 3 .

[0031] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, one of R 1 , R 2 is CH 3 , and the other is CHF 2 .

[0032] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 3 , R 4 , R 5 are each independently selected from CH 3 , i-Pr, OMe, CD 3 or halogen.

[0033] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 1 , R 2 respectively correspond to the groups or values shown by any compound in Table 1.

[0034] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 5 is CH 3 , and the CH 3 is optionally further substituted by 0 to 3 Ds; R 3 , R 4 are the same and are selected from CH 3, i-Pr, OMe or halogen.

[0035] In some specific embodiments of the present invention, in the compound of formula I or its pharmaceutically acceptable form of the present invention, R 3 , R 4 , R 5 correspond to the groups or values shown by any of the compounds in Table 1 respectively.

[0036] In some specific embodiments of the present invention, in the compound of formula I or its pharmaceutically acceptable form of the present invention, L is n is 0 or 1, preferably 0. Preferably, k is 0, 1 or 2. Preferably, R 9 , R 10 One of them is H, and the other is selected from H, C 1-6 linear alkyl, C 3-6 branched alkyl, C 3-6 cycloalkyl, C 3-6 heterocycloalkyl, C 6-10 aryl, C 5-10 heteroaryl and COOCH 3 ; the heteroaryl contains 1 or 2 heteroatoms N; the H in the L is optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, OH, =O, NH 2 , CN, COOH, C 1-4 alkyl, C 1-4 alkoxy.

[0037] In some specific embodiments of the present invention, in the compound of formula I or its pharmaceutically acceptable form of the present invention, L is

[0038] In some specific embodiments of the present invention, in the compound of formula I or its pharmaceutically acceptable form of the present invention, L is n is 0 or 1, k is 1.

[0039] In some specific embodiments of the present invention, in the compound of formula I or its pharmaceutically acceptable form of the present invention, L is n is 0 or 1, k is 1.

[0040] In some specific embodiments of the present invention, in the compound of formula I or its pharmaceutically acceptable form of the present invention, L is n is 2, k is 2.

[0041] In some specific embodiments of the present invention, in the compound of formula I or its pharmaceutically acceptable form of the present invention, L is n is 1, k is 1 or 2.

[0042] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, L is n is 2 and k is 2.

[0043] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, L corresponds to the group or value shown by any compound in Table 1.

[0044] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 6 is wherein, R 11 is amino, and the amino group is optionally further substituted by 0, 1 or 2 substituents selected from F, Cl, Br, I, OH, ═O, NH 2 , CN, COOH, CF 3 , C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl.

[0045] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 6 is wherein, R 11 is C 6-10 aryl or C 5-10 heterocyclic group, and the heterocyclic group contains 1, 2 or 3 heteroatoms N; the aryl group and heteroaryl group are optionally further substituted by 0, 1, 2 or 3 substituents selected from H, F, Cl, Br, I, OH, ═O, NH 2 , CN, COOH, CF 3 , C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl.

[0046] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 6 corresponds to the group or value shown by any compound in Table 1.

[0047] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 7 is H or CH 3 , and R 8 is H or F.

[0048] In some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 7 , R 8The group or value shown by any of the compounds in Table 1.

[0049] According to some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , and L respectively correspond to the group or value shown by any of the compounds in Table 1.

[0050] According to some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, the compound is selected from one or more of the compounds shown in Table 1:

[0051] Table 1

[0052]

[0053]

[0054]

[0055]

[0056]

[0057] According to some specific embodiments of the present invention, in the compound of formula I of the present invention or its pharmaceutically acceptable form, the pharmaceutically acceptable form is selected from pharmaceutically acceptable salts or co-crystals, stereoisomers, tautomers, deuterated compounds, solvates, chelates, non-covalent complexes or prodrugs.

[0058] <Second aspect>

[0059] The present invention also provides an intermediate compound having the structure shown in formula II:

[0060]

[0061] Wherein, R 1 , R 2 , R 3 , R 4 , R 5 , R 7 , R 8 As defined in any of the embodiments of the compound of formula I or its pharmaceutically acceptable form according to the <First aspect> of the present invention;

[0062] L 1 is selected from R 9 、R 10 、n, k are defined as in any one of the compounds of Formula I or their pharmaceutically acceptable forms according to the present invention; R 12 、R 13 are each independently H, Boc, Cbz, SEM, Fmoc, Alloc, Pht, OTs, PMB, Bn, Trt.

[0063] According to some specific embodiments of the present invention, the intermediate compounds of the present invention have the following structures:

[0064]

[0065] R 12 、R 13 One of them is H, and the other is Boc, Cbz, SEM, Fmoc, Alloc, Pht, OTs, PMB, Bn, Trt.

[0066] <Third aspect>

[0067] The present invention also provides a method for preparing the compound or its pharmaceutically acceptable form described in <First aspect> of the present invention. According to the chemical structure of the compound of Formula I or its pharmaceutically acceptable form, a synthetic route can be designed with reference to the known methods in the art to prepare the compound of Formula I or its pharmaceutically acceptable form.

[0068] According to some specific embodiments of the present invention, the method for preparing the compound or its pharmaceutically acceptable form described in <First aspect> of the present invention includes:

[0069] Performing a modification reaction on the carboxyl terminus shown in the intermediate compound of Formula II described in <Second aspect> of the present invention to prepare a compound having the structural formula I.

[0070] According to some specific embodiments of the present invention, the method for preparing the compound or its pharmaceutically acceptable form described in <First aspect> of the present invention further includes the process of preparing the intermediate compound described in <Second aspect> of the present invention.

[0071] According to some specific embodiments of the present invention, the method for preparing the compound or its pharmaceutically acceptable form described in <First aspect> of the present invention includes the steps shown in any one of the reaction routes in Examples 1 - 38.

[0072] According to some specific embodiments of the present invention, the method for preparing the compound or its pharmaceutically acceptable form described in <First aspect> of the present invention includes:

[0073]

[0074] <Fourth aspect>

[0075] The present invention also provides a pharmaceutical composition, which comprises: the compound according to the <First aspect> of the present invention or a pharmaceutically acceptable form thereof (preferably a pharmaceutically acceptable salt), and a pharmaceutically acceptable carrier, excipient and / or one or more other therapeutic agents.

[0076] <Fifth aspect>

[0077] The present invention also provides the use of the compound according to the <First aspect> of the present invention or a pharmaceutically acceptable form thereof (preferably a pharmaceutically acceptable salt) or the pharmaceutical composition according to the <Fourth aspect> of the present invention in the preparation of a preparation for inhibiting phosphodiesterase. Preferably, the phosphodiesterase includes PDE3 and / or PDE4.

[0078] <Sixth aspect>

[0079] The present invention also provides the use of the compound according to the <First aspect> of the present invention or a pharmaceutically acceptable form thereof (preferably a pharmaceutically acceptable salt) or the pharmaceutical composition according to the <Fourth aspect> of the present invention in the preparation of a drug for treating phosphodiesterase-related diseases.

[0080] The present invention also provides a method for treating phosphodiesterase-related diseases, which comprises administering to a subject an effective amount of the compound according to the <First aspect> of the present invention or a pharmaceutically acceptable form thereof (preferably a pharmaceutically acceptable salt) or the pharmaceutical composition according to the <Fourth aspect> of the present invention.

[0081] According to some specific embodiments of the present invention, the phosphodiesterase includes PDE3 and / or PDE4.

[0082] According to some specific embodiments of the present invention, the phosphodiesterase-related diseases include respiratory diseases such as asthma.

[0083] According to some specific embodiments of the present invention, the subject is a mammal or a human, preferably, the subject is a human.

[0084] The compound having the structural formula I of the present invention or a pharmaceutically acceptable form thereof can be used as a phosphodiesterase inhibitor, has high inhibitory activity against phosphodiesterase, especially PDE3 and / or PDE4, and has practical value.

[0085] Definitions and explanations

[0086] Unless otherwise indicated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered indefinite or unclear in the absence of a particular definition, but should be understood according to its ordinary meaning. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0087] Unless otherwise indicated, a solid wedge bond and a dashed wedge bond are used to represent the absolute configuration of a stereocenter, and a solid straight bond and a dashed straight bond are used to represent the relative configuration of a stereocenter, and a wavy line is used to represent a solid wedge bond or a dashed wedge bond or a wavy line is used to represent a solid straight bond and a dashed straight bond

[0088] The term “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0089] The term “pharmaceutically acceptable salt” or “its pharmaceutically acceptable salt” refers to salts of the compounds of the present invention, prepared from the compounds of the present invention having specific substituents with relatively non-toxic acids or bases. When a compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of a base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts or similar salts. When a compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of an acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, where the inorganic acids include, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, hydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, where the organic acids include, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid and similar acids; also salts of amino acids (such as arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and can thus be converted into either base or acid addition salts.

[0090] The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compounds containing acid radicals or basic groups by conventional chemical methods. Generally, the preparation method of such salts is to react these compounds in the form of free acids or bases with a stoichiometric amount of appropriate bases or acids in water, an organic solvent, or a mixture of both.

[0091] The term "eutectic" refers to a crystalline material that contains two or more distinct solids at room temperature, each solid having different physical properties, such as structure, melting point, and heat of fusion.

[0092] The term "stereoisomer" (or "optical isomer") refers to a stable isomer that has a perpendicular plane of asymmetry due to the presence of at least one chiral element (including chiral centers, chiral axes, chiral planes, etc.), enabling it to rotate plane-polarized light. Due to the presence of asymmetric centers and other chemical structures in the compounds of the present invention that may lead to stereoisomerism, the present invention also includes these stereoisomers and their mixtures. Since the compounds and their salts of the present invention may contain asymmetric carbon atoms, they can exist in the form of a single stereoisomer, a racemate, a mixture of enantiomers, and a mixture of diastereoisomers. Generally, these compounds can be prepared in the form of a racemic mixture. However, if desired, such compounds can be prepared or separated to obtain pure stereoisomers, i.e., single enantiomers or diastereoisomers, or mixtures enriched in a single stereoisomer (purity ≥ 98%, purity ≥ 95%, ≥ 93%, ≥ 90%, ≥ 88%, ≥ 85%, or ≥ 80%). The single stereoisomers of the compounds are synthesized from optically active starting materials containing the desired chiral centers, or are prepared by separating or resolving the mixture of enantiomeric products obtained, such as converting to a mixture of diastereoisomers and then separating or recrystallizing, chromatographic treatment, using chiral resolving agents, or directly separating the enantiomers on a chiral chromatographic column. The starting compounds with specific stereochemistry can be obtained commercially or prepared according to the methods described herein and then resolved by methods well known in the art. Unless otherwise indicated, all stereoisomeric forms of the compounds of the present invention are within the scope of the compounds of the present invention.

[0093] The term "tautomer" (or "tautomeric form") refers to structural isomers with different energies that can be interconverted through a low energy barrier. If tautomerism is possible (e.g., in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (or proton transfer tautomers) include (but are not limited to) interconversions carried out through proton migration, such as keto-enol isomerization, imine-enamine isomerization, amide-imidol isomerization, etc. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the compounds of the present invention.

[0094] Unless otherwise indicated, the compounds represented by the structural formulas of the present invention can be in the form of purified single stereoisomers or tautomeric forms, or in the form of mixtures containing multiple stereoisomers or tautomeric forms.

[0095] The term "solvate" refers to a substance formed by the compound of the present invention or its pharmaceutically acceptable salt in combination with at least one solvent molecule through non-covalent intermolecular forces. Common solvates include (but are not limited to) hydrates, ethanolates, acetonates, etc.

[0096] The term "chelate" is a coordination compound with a cyclic structure, obtained through the chelation of two or more ligands with the same metal ion to form a chelate ring.

[0097] The term "non-covalent complex" is formed through the interaction of a compound with another molecule, where no covalent bond is formed between the compound and the molecule. For example, complexation can occur through van der Waals interactions, hydrogen bonding, and electrostatic interactions (also known as ionic bonding).

[0098] The term "prodrug" refers to a derivative compound that can directly or indirectly provide the compound of the present invention after being administered to a patient. Particularly preferred derivative compounds or prodrugs are compounds that can improve the bioavailability of the compound of the present invention (e.g., are more easily absorbed into the blood) when administered to a patient, or compounds that promote the delivery of the parent compound to the site of action (e.g., the lymphatic system). Unless otherwise indicated, all prodrug forms of the compounds of the present invention are within the scope of the present invention, and various prodrug forms are well-known in the art.

[0099] The term "each independently" means that at least two groups (or ring systems) with the same or similar range of values present in a structure can have the same or different meanings in a specific situation. For example, if substituents X and Y are each independently hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl, then when substituent X is hydrogen, substituent Y can be either hydrogen or halogen, hydroxyl, cyano, alkyl, or aryl; similarly, when substituent Y is hydrogen, substituent X can be either hydrogen or halogen, hydroxyl, cyano, alkyl, or aryl.

[0100] The terms "comprising" and "including" are used in their open, non-limiting sense.

[0101] "Optional" or "optionally" means that the subsequent described event or circumstance may but does not necessarily occur, and this description includes the case where the described event or circumstance occurs and the case where the described event or circumstance does not occur.

[0102] The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by substituents, which may include deuterium and variants of hydrogen, as long as the valence state of the specific atom is normal and the resulting compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the type and number of substituents can be arbitrary based on what is chemically achievable.

[0103] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0 - 2 Rs, the group may optionally be substituted by up to two Rs, and each R has an independent option in each case. In addition, combinations of substituents and / or their variants are only permitted if such combinations result in a stable compound.

[0104] When a variable selected from a structural formula is missing, it means its absence. For example, when R is selected as missing in C - R, it means the structure is actually C.

[0105] When a variable connecting two groups in a structural formula is selected from a bond or absent, it means the two groups it connects are directly linked. For example, in A - L - Z, when L represents a bond or is absent, it means the structure is actually A - Z.

[0106] When it is not specified which atom of an enumerated substituent is bonded to the group being substituted, such a substituent can be bonded through any of its atoms. For example, a phenyl group as a substituent can be bonded to the group being substituted through any carbon atom on the benzene ring.

[0107] Unless otherwise specified, the term "alkyl" is used to denote a straight-chain or branched-chain saturated hydrocarbon group, which can be mono-substituted (e.g., -CH 2 F) or poly-substituted (e.g., -CF 3) can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). Examples of alkyl groups include methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, s-butyl, t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), etc.

[0108] Unless otherwise specified, the term "alkylene" refers to a divalent straight-chain or branched-chain alkane group consisting only of carbon and hydrogen atoms, having no unsaturation, and connected to other moieties by two single bonds, including (but not limited to) methylene, 1,1-ethylene, and 1,2-ethylene, etc. For example, "C 1-3 alkylene" refers to a saturated divalent straight-chain or branched-chain alkyl group containing 1 to 3 carbon atoms.

[0109] Unless otherwise specified, "cycloalkyl" includes any stable cyclic or polycyclic hydrocarbon group, any carbon atom of which is saturated, and which can be mono-substituted or poly-substituted, and can be monovalent, divalent or polyvalent. Examples of these cycloalkyl groups include, but are not limited to, cyclopropyl, norbornyl, [2.2.2]bicyclooctane, [4.4.0]bicyclodecane, etc.

[0110] Unless otherwise specified, the term "alkoxy" means that an alkyl group is connected to the rest of the molecule through an oxygen atom, where the alkyl group has the meaning as described in the present invention. Unless otherwise specified, C 1-5 alkoxy includes C1, C2, C3, C4 and C5 alkoxy. Examples of alkoxy include but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentyloxy and S-pentyloxy. The alkoxy group can optionally be substituted by one or more substituents described in the present invention.

[0111] Unless otherwise specified, the term "3- to 6-membered ring" means a saturated or unsaturated monocyclic ring containing or not containing heteroatoms, having 3, 4, 5 or 6 C, O, S, N atoms in the ring; the "3- to 6-membered ring" can be connected to the rest of the structural formula through any carbon atom, or, if present, a nitrogen atom.

[0112] Unless otherwise specified, the term "amino group" means -NH 2 , -NH(alkyl) or -N(alkyl)(alkyl).

[0113] Unless otherwise specified, the term "aromatic ring" means a polyunsaturated aromatic alkane monocyclic ring, which can be mono-substituted or poly-substituted.

[0114] Unless otherwise specified, the term "4-6-membered heteroalkyl" refers to a saturated monovalent monocyclic hydrocarbon ring having 3, 4 or 5 carbon atoms in the ring and one or more heteroatom groups selected from O, NR a , where R a represents a hydrogen atom or C 1-6 alkyl; said "4-6-membered heteroalkyl" can be attached to the rest of the molecule through any carbon atom, or, if present, a nitrogen atom.

[0115] Unless otherwise specified, the term "heteroaryl ring" refers to an aromatic ring containing one to four heteroatoms selected from one or more of N, O and S.

[0116] Unless otherwise specified, the term "heterocyclic group" refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic, e.g., fused ring, bridged ring or spiro ring) non-aromatic group, the ring atoms of which consist of carbon atoms and at least one heteroatom selected from N, O and S, where the S atom is optionally substituted to form S(=O), S(=O) 2 or S(=O)(=NR x ), R x independently selected from H or C 1-4 alkyl. If the valence bond requirements are met, the heterocyclic group can be attached to the rest of the molecule through any ring atom. For example, the term "3-8-membered heterocyclic group" used in the present invention refers to a heterocyclic group having 3 to 8 ring atoms. For example, the heterocyclic group can be oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuranyl, dioxolanyl, pyrrolidinyl, pyrrolidinone, imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, dithianyl or trithianyl.

[0117] Unless otherwise specified, the term "aryl" refers to a monocyclic or fused polycyclic aromatic hydrocarbon group having a conjugated π electron system. For example, the term "C 6-10 aryl" used in the present invention refers to an aryl having 6 to 10 carbon atoms. For example, aryl can be phenyl, naphthyl, anthracenyl, phenanthryl, acenaphthylenyl, azulyl, fluorenyl, indenyl, pyrenyl, etc.

[0118] Unless otherwise specified, the term "heteroaryl" refers to a monocyclic or fused polycyclic aromatic group having a conjugated π-electron system, the ring atoms of which are composed of carbon atoms and at least one heteroatom selected from N, O, and S. If the valence bond requirements are satisfied, the heteroaryl can be linked to the rest of the molecule through any one of the ring atoms. For example, the term "5- to 10-membered heteroaryl" as used in the present invention refers to a heteroaryl having 5 to 10 ring atoms. For example, the heteroaryl can be thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl and their benzo derivatives, pyrrolopyridyl, pyrrolopyrazinyl, pyrazolopyridyl, imidazolopyridyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl, purinyl, etc.

[0119] Unless otherwise specified, the term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I). The term "hydroxyl" refers to -OH. The term "cyano" refers to -CN. The term "amino" refers to -NH 2 。

[0120] Unless otherwise specified, the term "pharmaceutical composition" refers to a mixture formed by one or more compounds described in the present invention or pharmaceutically acceptable forms and other chemical components, wherein the "other chemical components" refer to pharmaceutically acceptable carriers, excipients, and / or one or more other therapeutic agents. A "carrier" refers to a material that does not cause significant irritation to the organism and does not eliminate the biological activity and properties of the administered compound. An "excipient" refers to an inert substance added to the pharmaceutical composition to facilitate the administration of the compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, and disintegrants.

[0121] The compounds of the present invention have good PDE inhibitory activity and have potential application value in the treatment of diseases related to PDE, especially related to PDE3 / PDE4. Description of the Drawings

[0122] Figure 1 It is the single crystal diffraction pattern of Compound 1-2 of the present invention. Detailed Description of the Invention

[0123] The embodiments of the present invention will be described in detail below in conjunction with the examples. However, those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. For those not specified in the examples, they are carried out under conventional conditions or conditions recommended by the manufacturer. For those reagents or instruments not specified by the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0124] The structure of the compound was determined by nuclear magnetic resonance or mass spectrometry. Nuclear magnetic resonance was measured using a BRUKER 400M nuclear magnetic resonance instrument, and the measuring solvent was deuterated dimethyl sulfoxide (DMSO-d 6 ), or deuterated chloroform (CDCl 3 ), with tetramethylsilane (TMS) as the internal standard, and the chemical shift (δ) was given in units of 10 -6 (ppm). Mass spectrometry was measured using a Waters ACQUITY Arc / ACQUITY QDa or a Thermo U3000-ISQ EC liquid chromatography-mass spectrometry instrument.

[0125] High performance liquid chromatography analysis was performed using a Thermo U3000 high pressure liquid chromatography instrument. High performance liquid chromatography preparation was performed using a Hanbang DAC-50 or a Shimadzu LC-20AP preparative chromatograph.

[0126] Reaction monitoring was performed using thin layer chromatography or liquid chromatography-mass spectrometry. The developing agent systems used in thin layer chromatography were: dichloromethane and methanol system, petroleum ether and ethyl acetate system. The volume ratio of the solvents was adjusted according to the polarity of the compound, or a small amount of triethylamine was added for adjustment. Liquid chromatography-mass spectrometry was performed using a Waters ACQUITY Arc / ACQUITY QDa or a Thermo U3000-ISQ EC liquid chromatography-mass spectrometry instrument.

[0127] Column chromatography generally used silica gel with 200 - 300 mesh as the carrier. The eluent systems included: dichloromethane and methanol system, petroleum ether and ethyl acetate system. The volume ratio of the solvents was adjusted according to the polarity of the compound, or a small amount of triethylamine was added for adjustment.

[0128] Unless otherwise specified in the following examples and comparative examples, the reaction temperature was room temperature (20°C - 30°C), and the solvents were all dried and purified according to standard methods.

[0129] The compounds of the comparative examples are shown in Table 2.

[0130] Table 2. Compounds of the Comparative Examples

[0131]

[0132] Preparation of the compound D1 of the comparative example:

[0133]

[0134] Preparation steps: 1.

[0136]

[0137] Under nitrogen protection at room temperature, to a solution of 2,4,6-trimethylaniline (2.0 g, 14.7 mmol, 1 equiv) and tert-butyl N-(3-bromopropyl)carbamate (14.1 g, 59.1 mmol, 4 equiv) in DMF (20 mL, 258.4 mmol), K 2 CO 3 (6.1 g, 44.3 mmol, 3 equiv) and KI (2.5 g, 14.7 mmol, 1 equiv) were added respectively. The reaction was carried out overnight at 80 °C. The reaction mixture was extracted with EA (3 × 50 ml). The combined organic phases were washed with saturated sodium chloride solution (2 × 30 ml), dried over anhydrous sodium sulfate. The mixture was filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography, EA / PE (1 / 4), to give tert-butyl N-(3-[(2,4,6-trimethylphenyl)amino]propyl)carbamate (2.2 g, yield 50.86%).

[0138] LCMS (ESI, m / z): [M+H] + = 293.2 2.

[0140]

[0141] Under nitrogen protection at room temperature, to a solution of tert-butyl N-(3-[(2,4,6-trimethylphenyl)amino]propyl)carbamate (1.2 g, 4.1 mmol, 1 equiv) and 2-chloro-9,10-dimethoxy-6,7-dihydropyrimido[4,3-a]isoquinolin-4-one (1.8 g, 6.1 mmol, 1.5 equiv) in dioxane (20 mL), Cs 2 CO 3 (2.7 g, 8.2 mmol, 2 equiv), APhos Pd G 3 (0.3 g, 0.4 mmol, 0.1 equiv) and 4-(di-tert-butylphosphino)-N,N-dimethylaniline (0.1 g, 0.4 mmol, 0.1 equiv) were added respectively. Under nitrogen protection, the reaction was stirred overnight at 100 °C. The reaction mixture was extracted with EA (3 × 50 mL). The combined organic phases were washed with saturated sodium chloride solution (2 × 30 mL), dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse-phase column chromatography under the following conditions: C18 column, mobile phase, water and acetonitrile, 10% to 90% gradient for 30 minutes, UV: 254 nm. To give tert-butyl N-[3-((9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl)(2,4,6-trimethylphenyl)amino)propyl]carbamate (200 mg, yield 8.88%).

[0142] LCMS (ESI, m / z): [M+H] + = 549.4 3.

[0144]

[0145] Under nitrogen protection, at 0 °C, trifluoroacetic acid (TFA, 5 mL) was added dropwise to a solution of tert-butyl N-[3-((9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl)(2,4,6-trimethylphenyl)amino)propyl]carbamate (290 mg, 1 equivalent) in DCM (5 mL). Under nitrogen protection, the resulting residue was stirred at room temperature for 2 h. The reaction mixture was extracted with DCM (3 × 20 mL). The combined organic phases were washed with saturated sodium chloride solution (2 × 10 mL) and dried over anhydrous sodium sulfate. After filtration of the resulting mixture, the filtrate was concentrated under reduced pressure. 2-[(3-Aminopropyl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (200 mg, crude) was obtained.

[0146] LCMS (ESI, m / z): [M+H] + = 449.3 4.

[0148]

[0149] Under nitrogen protection, at 0 °C, trimethylsilyl isocyanate (128.4 mg, 1.1 mmol, 2.5 equivalents) was added dropwise to a solution of 2-[(3-aminopropyl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (200 mg, 0.4 mmol, 1.0 equivalent) and triethylamine (TEA, 135 mg, 1.3 mmol, 3 equivalents) in DCM (5 mL). Under nitrogen protection, the resulting mixture was stirred at 50 °C for 2 days. The resulting residue was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography to obtain 3-((9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl)(2,4,6-trimethylphenyl)amino)propylurea (7.5 mg, yield 3.4%), with the following conditions (column type: Xbridge BEH Phenyl 5 μm, 19 * 250 mm; mobile phase A: water (0.1% FA), mobile phase B: MeOH; flow rate: 25 mL / min, gradient: 43% B to 53% B in 10 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 9.6

[0150] LCMS (ESI, m / z): [M+H]+ = 492.25

[0151] 1 H NMR (400 MHz, Methanol-d 4 ) δ 8.22 (broad, 1H), 7.26 (s, 1H), 7.01 (s, 2H), 6.94 (s, 1H), 6.24 (s, 1H), 4.05 (t, J = 6.4 Hz, 2H), 3.91 (s, 6H), 3.61 (t, J = 7.8 Hz, 2H), 3.45 (t, J = 6.6 Hz, 2H), 3.01–2.89 (m, 2H), 2.29 (s, 3H), 2.24 (s, 6H), 1.88 (s, 2H).

[0152] Preparation of Comparative Compound D2:

[0153]

[0154] At room temperature, (2E)-9,10-dimethoxy-2-[(2,4,6-trimethylphenyl)imino]-3H, 6H, 7H-pyrimido[4,3-a]isoquinolin-4-one (1.0 g, 2.55 mmol, 1.0 equiv) and dichloromethane (20 mL) were added to a 50 mL flask and stirred. Acetic anhydride (0.29 g, 2.81 mmol, 1.1 equiv) was added, and the mixture was stirred at room temperature under nitrogen protection for 24 h. After the reaction was completed, the system was cooled to 0 °C, 10 ml of water was added dropwise, and the mixture was stirred for 1 h. The mixture was extracted with dichloromethane three times, the organic phases were combined, dried and evaporated, and purified by silica gel column to obtain compound N-(9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)-N-meta-methylacetamide (0.11 g).

[0155] LCMS (ESI, m / z): [M+H] + = 434.21

[0156] 1 H NMR (400 MHz, CDCl 3 ) δ 7.72 (s, 1H), 7.18 (s, 1H), 6.94 (s, 2H), 6.74 (s, 1H), 4.19 (t, J = 6.4 Hz, 2H), 3.92 (s, 6H), 2.95 (t, J = 6.4 Hz, 2H), 2.31 (s, 3H), 2.12 (s, 6H), 2.05 (s, 3H).

[0157] Preparation of Comparative Compound D3:

[0158]

[0159] (2E)-9,10-Dimethoxy-2-[(2,4,6-trimethylphenyl)imino]-3H,6H,7H-pyrimido[4,3-a]isoquinolin-4-one (1.00 g, 2.554 mmol, 1.0 eq), 2-bromopropane (0.47 g, 3.831 mmol, 1.5 eq), Pd 2 (dba) 3 (0.23 g, 0.255 mmol, 0.1 eq), Cs 2 CO 3 (2.50 g, 7.662 mmol, 3.0 eq) were dissolved in 1,4-dioxane (20 ml). Under nitrogen protection, the mixture was stirred and heated to reflux for 12 h. After the reaction was completed, water (50 ml) was added and the mixture was extracted with ethyl acetate (50 ml × 3), washed with saturated sodium chloride (50 ml × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 2-(isopropyl(methyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (7.8 g, 71.43%).

[0160] LCMS (ESI, m / z): [M+H] + = 434.25

[0161] 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.08 (s, 2H), 6.89 (s, 1H), 6.58 (s, 1H), 5.35 (s, 1H), 4.82–4.74 (m, 1H), 4.06 (t, J = 6.4 Hz, 2H), 3.86 (s, 3H), 3.63 (s, 3H), 2.93 (t, J = 6.4 Hz, 2H), 2.34 (s, 3H), 2.14 (s, 3H), 1.25 (s, 3H), 1.23 (s, 3H).

[0162] Preparation of Comparative Compound D4:

[0163]

[0164] 9,10 - Dimethoxy - 2 - [(2,4,6 - trimethylphenyl)amino] - 6H,7H - pyrimido[4,3 - a]isoquinolin - 4 - 1 (200 mg, 0.5 mmol, 1.0 equiv) was dissolved in the solvent DMF (2 mL). NaH (40 mg, 1.0 mmol, 2.0 equiv, 60% purity) was added at 0 °C, and the resulting mixture was stirred under nitrogen protection for 0.5 h. (2 - Bromoethyl)diethylammonium hydrobromide (400 mg, 1.5 mmol, 3.0 equiv) was added to the above mixture, and the mixture was stirred at room temperature for 1 h. The reaction was quenched by adding water (10 mL) at room temperature. The mixture was extracted with ethyl acetate (2 × 50 mL), and the organic layer was washed with saturated brine (3 × 100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography (EA / PE = 1 / 2) gave 2 - {[2 - (diethylamino)ethyl](2,4,6 - trimethylphenyl)amino} - 9,10 - dimethoxy - 6H,7H - pyrimido[4,3 - a]isoquinolin - 4 - 1 (37.2 mg, yield 14%).

[0165] LCMS(ESI): [M + H] + = 491.25

[0166] 1 H NMR(400 MHz, Methanol - d 4 ) δ 6.95–6.85 (m, 3H), 6.77 (s, 1H), 5.50 (s, 1H), 4.44–4.35 (m, 2H), 4.06–3.99 (m, 2H), 3.87 (s, 3H), 3.69 (s, 3H), 2.99–2.90 (m, 4H), 2.74 (q, J = 7.2 Hz, 4H), 2.28 (s, 3H), 2.05 (s, 6H), 1.16 (t, J = 7.2 Hz, 6H).

[0167] Preparation of Comparative Compound D5:

[0168]

[0169] Preparation steps: 1.

[0171]

[0172] Under nitrogen protection, at room temperature, 9,10 - dimethoxy - 2 - [(2,4,6 - trimethylphenyl)amino] - 6H,7H - pyrimido[4,3 - a]isoquinolin - 4 - one (500 mg, 1.2 mmol, 1.0 equiv) and Cs 2 CO 3(1248 mg, 3.8 mmol, 3.0 equiv) was added to a solution of DMF (10 mL), and N-(2-bromoethyl)carbamic acid tert-butyl ester (715 mg, 3.1 mmol, 2.5 equiv) and Pd 2 (dba) 3 (116 mg, 0.1 mmol, 0.1 equiv) were added portionwise. Under nitrogen protection, the resulting residue was stirred at 100 °C overnight. The reaction mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic phases were washed with saturated sodium chloride solution (2 × 30 mL) and dried over anhydrous sodium sulfate. After filtration of the resulting mixture, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography, dichloromethane / methanol (20 / 1), to give tert-butyl N-[2-((9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl)(2,4,6-trimethylphenyl)amino)ethyl]carbamate (200 mg, 29.29%).

[0173] LCMS (ESI): [M+H] + = 535.2 2.

[0175]

[0176] Under nitrogen protection, at room temperature, tert-butyl N-[2-((9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl)(2,4,6-trimethylphenyl)amino)ethyl]carbamate (180 mg, 0.3 mmol, 1.0 equiv) was added to a dioxane solution of HCl (4 mL, 4 M), and the mixture was stirred for 1 h. The reaction mixture was basified to pH = 8 with saturated aqueous sodium bicarbonate. The reaction mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic phases were backwashed with saturated sodium chloride solution (2 × 10 mL) and dried over anhydrous sodium sulfate. After filtration of the resulting mixture, the filtrate was concentrated under reduced pressure. 2-[(2-Aminoethyl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (160 mg, crude product) was obtained.

[0177] LCMS (ESI): [M+H] + = 435.2 3.

[0179]

[0180] Under nitrogen protection, at room temperature, 2-[(2-aminoethyl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (100 mg, 0.2 mmol, 1.0 equivalent) and TEA (69 mg, 0.6 mmol, 3.0 equivalents) were added to a DCM (5 mL) solution, and isocyano trimethylsilane (53 mg, 0.4 mmol, 2.0 equivalents) was added dropwise. Under nitrogen protection, at room temperature, the resulting residue was stirred and reacted for 1.5 h. The reaction mixture was extracted with dichloromethane (3X 20 mL). The combined organic phases were backwashed with saturated brine (2X 50 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography to obtain 2-((9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl(2,4,6-trimethylphenyl)amino)ethyl urea (26.4 mg, 24.02%), with the following conditions (column type: YMC Triart C18 ExRs 5μm, 19mm*250mm; mobile phase A: water (10mmol / L NH 4 HCO 3 ), mobile phase B: acetonitrile; flow rate: 25 mL / min; gradient: 32% B to 55% B in 10 min; wavelength: 254 nm / 220 nm; RT1 (min): 9.17).

[0181] LCMS (ESI): [M+H] + = 478.20

[0182] 1 1H NMR (400 MHz, Methanol-d 4 ) δ 7.11 (s, 2H), 6.94 (s, 1H), 6.70 (s, 1H), 5.51 (s, 1H), 4.15–4.08 (m, 2H), 3.99–3.92 (m, 2H), 3.89 (s, 3H), 3.67 (s, 3H), 3.44 (t, J = 6.4 Hz, 2H), 2.97 (t, J = 6.4 Hz, 2H), 2.36 (s, 3H), 2.18 (s, 6H).

[0183] Preparation of Comparative Compound D6:

[0184]

[0185] (2E)-9,10-dimethoxy-2-[(2,4,6-trimethylphenyl)imino]-3H,6H,7H-pyrimido[4,3-a]isoquinolin-4-one (500 mg, 1.28 mmol, 1.0 eq), 1-bromobutane (263 mg, 1.92 mmol, 1.5 eq), Pd 2 (dba) 3 (119 mg, 0.13 mmol, 0.1 eq), Cs 2 CO 3 (1.25 g, 3.84 mmol, 3.0 eq), Xantphos (150 mg, 26 mmol, 0.2 eq) were dissolved in 1,4-dioxane (10 ml). Under nitrogen protection, the mixture was stirred and heated to reflux for 12 h. After the reaction was completed, water (50 ml) and ethyl acetate were added for extraction (50 ml × 3), washed with saturated sodium chloride (50 ml × 1), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain 2-(butyl(m-tolyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (121 mg, 21.1%).

[0186] LCMS(ESI): [M+H] + = 448.26

[0187] 1 H NMR (400 MHz, CDCl 3 ) δ 7.01 (s, 2H), 6.71 (s, 1H), 6.64 (s, 1H), 5.34 (s, 1H), 4.17 (t, J = 6.4 Hz, 2H), 3.93–3.85 (m, 5H), 3.73 (s, 3H), 2.90 (t, J = 6.4 Hz, 2H), 2.35 (s, 3H), 2.16 (s, 6H), 1.62–1.51 (m, 2H), 1.40–1.27 (m, 2H), 0.89 (t, J = 8.0 Hz).

[0188] Example 1, Compound 1-1 and Compound 1-2

[0189]

[0190] Preparation steps: 1.

[0192]

[0193] tert-Butyl N-(2-bromopropyl)carbamate (0.91 g, 3.8 mmol, 1.5 equiv), cesium carbonate (2.50 g,, 7.662 mmol, 3 equiv), Pd2 (dba) 3 (0.23 g, 0.26 mmol, 0.1 eq) and (E)-2-(Methylsulfinylimino)-9,10-dimethoxy-2,3,6,7-tetrahydro-4H-pyrimido[6,1-a]isoquinolin-4-one (1 g, 2.54 mmol, 1.0 eq) were dissolved in 20 mL of 1,4-dioxane under nitrogen protection and stirred at 110 °C overnight. Concentrated under reduced pressure, dissolved in 10 mL of water, extracted with ethyl acetate (3 × 10 mL), the organic layer was washed with 20 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse-phase column chromatography under the following conditions (column: C18; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 30% B to 50% B in 20 min; wavelength: 254 nm;) to obtain tert-butyl (E)-(2-(2-(methylsulfinylimino)-9,10-dimethoxy-4-oxo-6,7-dihydro-2H-pyrimido[6,1-a]isoquinolin-3(4H)-yl)propyl)carbamate (500 mg, yield 35.6%).

[0194] LCMS(ESI): [M+H] + = 549.06 2.

[0196]

[0197] tert-Butyl (E)-(2-(2-(methylsulfinylimino)-9,10-dimethoxy-4-oxo-6,7-dihydro-2H-pyrimido[6,1-a]isoquinolin-3(4H)-yl)propyl)carbamate (500 mg, 0.91 mmol, 1 eq) was dissolved in a 1,4-dioxane solution of 10 ml of hydrochloric acid (4 M) and stirred at room temperature for 1 hour. Concentrated under reduced pressure, dissolved in 10 mL of water, extracted with ethyl acetate (3 × 10 mL), the organic layer was washed with 20 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 2-((1-aminopropan-2-yl)(methylsulfonyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (300 mg, yield 73.3%).

[0198] LCMS(ESI): [M+H] + = 449.16 3.

[0200]

[0201] 2-((1-Aminopropan-2-yl)(methylsulfonyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (200 mg, 0.45 mmol, 1.0 equiv.), triethylamine (135 mg, 1.34 mmol, 3 equiv.) and trimethylsilyl isocyanate (102.73 mg, 0.9 mmol, 2 equiv.) were dissolved in 10 mL of dichloromethane and stirred at room temperature for 2 h. Concentrated under reduced pressure, dissolved in 10 mL of water, and extracted with ethyl acetate (3 × 10 mL). The organic layer was washed with 20 mL of saturated sodium chloride and dried over anhydrous Na 2 SO 4 and filtered, then concentrated under reduced pressure. The crude product was purified by reverse-phase column chromatography under the following conditions (column: Xselect CSH C18 5 μm, 30 mm X 150 mm; mobile phase A: water (0.1% FA), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 28% B to 38% B in 8 min; wavelength: 254 nm / 220 nm; RT1 (min): 5.85 / 7.65) to obtain 1-(2-((9,10-Dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)(methylsulfonamido)propyl)urea (100 mg, yield 45.6%).

[0202] LCMS (ESI): [M+H] + = 492.35 4.

[0204]

[0205] The racemate (100 mg) was purified by Chiral-HPLC under the conditions of column: CHIRALPAK IE-3, 4.6*50 mm, 3 μm; mobile phase A: DCM--HPLC, mobile phase B: ETOH:DCM = 1:1; flow rate: 18 mL / min; Gradient: isocratic 20; wavelength: 220 nm; RT1 (min): 17.4; RT2 (min): 21.8; sample solvent: MeOH-HPLC; injection volume: 0.45 mL; number of runs: 5. The product was obtained.

[0206] Compound 1-1 (isomer 1, RT1 (min): 17.4, 13.8 mg, yield 22.14%)

[0207] LCMS (ESI): [M+H] + = 492.35

[0208] 1¹H NMR (400 MHz, Methanol-d 4 ) δ 7.14–7.11 (d, J=10.1 Hz, 2H), 6.93 (s, 1H), 6.64 (s, 1H), 5.40 (s, 1H), 4.49 (broad, 1H), 4.20–4.04 (m, 2H), 3.89 (s, 3H), 3.66 (s, 3H), 2.97 (t, J=6.5 Hz, 2H), 2.37 (s, 3H), 2.22 (s, 3H), 2.10 (s, 3H), 1.05 (s, 3H).

[0209] Compound 1-2 (isomer 2, RT2 (min): 21.8, 13.0 mg, yield 22.0%)

[0210] LCMS (ESI): [M+H] + =492.35

[0211] 1 ¹H NMR (400 MHz, Methanol-d 4 ) δ 7.12 (d, J=10.1 Hz, 2H), 6.93 (s, 1H), 6.64 (s, 1H), 5.40 (s, 1H), 4.49 (broad, 1H), 4.20–4.04 (m, 2H), 3.89 (s, 3H), 3.66 (s, 3H), 2.97 (t, J=6.5 Hz, 2H), 2.37 (s, 3H), 2.22 (s, 3H), 2.08 (s, 6H), 1.05 (s, 3H).

[0212] The single crystal diffraction pattern of Compound 1-2 is as Figure 1 .

[0213] Example 2, Compound 2

[0214]

[0215] Preparation steps: 1.

[0217]

[0218] Dissolve 9,10 - dimethoxy - 2 - [(2,4,6 - trimethylphenyl)amino]-6H,7H - pyrimido[4,3 - a]isoquinolin - 4 - one (3.0 g, 7.7 mmol, 1.0 equiv) and tert - butyl 3 - bromopyrrolidine - 1 - carboxylate (2.3 g, 9.2 mmol, 1.2 equiv) in 1,4 - dioxane (100 mL), add Xantphos (0.9 g, 1.5 mmol, 0.2 equiv), cesium carbonate (5.0 g, 15.3 mmol, 2.0 equiv) and Pd 2 (dba) 3 (0.7 g, 0.7 mmol, 0.1 equiv). Stir the mixture under a nitrogen atmosphere at 100 °C for 18 h, then concentrate the resulting mixture under reduced pressure. The residue is purified by reverse phase with the following conditions: column, C18 silica gel column; mobile phase, acetonitrile, water, 0% - 100% gradient, 30 minutes; detector, UV 254 nm, to obtain tert - butyl 3 - ({9,10 - dimethoxy - 4 - oxo - 6H,7H - pyrimido[4,3 - a]isoquinolin - 2 - yl}(2,4,6 - trimethylphenyl)amino)pyrrolidine - 1 - carboxylate (1.4 g, yield 32.6%).

[0219] LCMS(ESI): [M + H] + = 561.3 2.

[0221]

[0222] Mix tert - butyl 3 - ({9,10 - dimethoxy - 4 - oxo - 6H,7H - pyrimido[4,3 - a]isoquinolin - 2 - yl}(2,4,6 - trimethylphenyl)amino)pyrrolidine - 1 - carboxylate (300 mg, 0.5 mmol, 1.0 equiv) with a 1,4 - dioxane solution of hydrochloric acid (8 mL), and stir the resulting mixture at room temperature for 1 h. Concentrate the resulting mixture under reduced pressure to obtain 9,10 - dimethoxy - 2 - [pyrrolidin - 3 - yl(2,4,6 - trimethylphenyl)amino]-6H,7H - pyrimido[4,3 - a]isoquinolin - 4 - one (300 mg, crude). The crude product is used directly in the next step without further purification.

[0223] LCMS(ESI): [M + H] + = 461.2 3.

[0225]

[0226] At room temperature, 9,10-dimethoxy-2-[(pyrrolidin-3-yl)(2,4,6-trimethylphenyl)amino]-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (100 mg, 0.2 mmol, 1.0 equiv) was dissolved in dichloromethane (4 mL), and triethylamine (109 mg, 1.1 mmol, 5.0 equiv) and isocyano trimethylsilane (38 mg, 0.3 mmol, 1.5 equiv) were added. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The crude product was purified by reverse phase (column: YMC-Triart C8 5.5 mmol / L, mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: from 32% B to 50% B in 7 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 6.23), to give 3-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)pyrrolidine-1-carboxamide (30.5 mg, yield 27.4%).

[0227] LCMS(ESI,m / z):[M+H] + =504.25

[0228] 1 H NMR(400MHz,Methanol-d4)δ7.13(s,2H),6.92(s,1H),6.64(s,1H),5.41(s,1H),5.08–4.99(m,1H),4.19–4.03(m,3H),3.88(s,3H),3.66(s,3H),3.58–3.52(m,1H),3.29(t,J=7.3Hz,1H),3.25–3.18(m,1H),2.96(t,J=6.5Hz,2H),2.38(s,3H),2.29–2.22(m,1H),2.17–2.15(m,6H),1.77(s,1H).

[0229] Example 3, Compound 3-1 and Compound 3-2

[0230]

[0231] Preparation steps: 1.

[0233]

[0234] Add 3,4 - dimethoxyphenethylamine (50.0 g, 275.8 mmol, 1.0 equiv) and ethyl cyanoacetate into a single - necked flask successively. Under nitrogen protection, react at 100 °C for 16 h, then cool down to 70 °C and add ethanol (80 mL). Filter, wash the filter cake with ethanol (3 x 30 mL), and dry the residual ethanol in the filter cake to obtain 2 - cyano - N - [2 - (3,4 - dimethoxyphenyl)ethyl]acetamide (50.0 g, 73.0% yield).

[0235] LCMS(ESI,m / z):[M + H] + =249.2 2.

[0237]

[0238] Add 2 - cyano - N - [2 - (3,4 - dimethoxyphenyl)ethyl]acetamide (50.0 g, 201.3 mmol, 1.0 equiv) and phosphorus oxychloride (500 mL) into a single - necked flask successively. Under nitrogen protection, react at 85 °C overnight. Dry the solvent, add water (300 mL), extract with DCM (3 x 400 mL), dry the combined organic phase with anhydrous sodium sulfate, and dry the organic phase to obtain 2 - [(1E) - 6,7 - dimethoxy - 3,4 - dihydro - 2H - isoquinolin - 1 - ylidene]acetonitrile (45.0 g, 97.0%).

[0239] LCMS(ESI,m / z):[M + H] + =231.2 3.

[0241]

[0242] Add 2 - [(1E) - 6,7 - dimethoxy - 3,4 - dihydro - 2H - isoquinolin - 1 - ylidene]acetonitrile (45.0 g, 195.4 mmol, 1.0 equiv) and concentrated sulfuric acid (320 mL) into a single - necked flask successively. React at room temperature for 3 h. Slowly pour the reaction solution into ice - water (1 L), adjust the pH to 7 with sodium hydroxide (4 M) at 0 °C, extract with EtOAc (3 x 600 mL), dry the combined organic phase with anhydrous sodium sulfate, and dry the organic phase to obtain 2 - [(1E) - 6,7 - dimethoxy - 3,4 - dihydro - 2H - isoquinolin - 1 - ylidene]acetamide (40.0 g, 82.4%).

[0243] LCMS(ESI,m / z):[M + H] + =249.2 4.

[0245]

[0246] In a single-necked flask, 2-[(1E)-6,7-dimethoxy-3,4-dihydro-2H-isoquinolin-1-ylidene]acetamide (40.0 g, 161.1 mmol, 1.0 equiv), ethanol (600 mL), and an ethanol solution of sodium ethoxide (127 mL, 21%) were successively added. The reaction was carried out at 80 °C for 0.5 h, and then diethyl carbonate (57.1 g, 483.3 mmol, 3.0 equiv) was added at this temperature. The reaction was carried out overnight at 80 °C. After cooling to room temperature, water (500 mL) was added, and the mixture was acidified to pH = 7 with dilute hydrochloric acid. The mixture was filtered, and the filter cake was washed with ethanol (3 × 100 mL). The ethanol remaining in the filter cake was evaporated to obtain 9,10-dimethoxy-3H,6H,7H-pyrimido[4,3-a]isoquinoline-2,4-dione (40.0 g, 90.5%).

[0247] LCMS(ESI,m / z):[M+H] + =275.2 5.

[0249]

[0250] In a single-necked flask, 9,10-dimethoxy-3H,6H,7H-pyrimido[4,3-a]isoquinoline-2,4-dione (40.0 g, 145.8 mmol, 1.0 equiv) and phosphorus oxychloride (240 mL) were successively added. Under nitrogen protection, the reaction was carried out overnight at 100 °C. The solvent was evaporated, water (500 mL) was added for dissolution, and the mixture was extracted with DCM (3 × 500 mL). The combined organic phases were dried over anhydrous sodium sulfate, and the organic phases were evaporated to obtain 2-chloro-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (30.0 g, 70.2%).

[0251] LCMS(ESI,m / z):[M+H] + =293.2 6.

[0253]

[0254] In a single-necked flask, 2-chloro-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (2.0 g, 6.8 mmol, 1.0 equiv), isopropanol (50 mL), and trimethylaniline (4.6 g, 34.1 mmol, 5.0 equiv) were added successively. Under nitrogen protection, the reaction was carried out overnight at 90 °C. The solvent was evaporated to dryness, and EA (20 mL) and saturated aqueous sodium bicarbonate solution (200 mL) were added. The mixture was filtered, and the filter cake was washed with EA (3 x 50 mL). The residual EA in the filter cake was evaporated to dryness to obtain (2E)-9,10-dimethoxy-2-[(2,4,6-trimethylphenyl)imino]-3H,6H,7H-pyrimido[4,3-a]isoquinolin-4-one (2.0 g, 74.8%).

[0255] LCMS(ESI,m / z):[M+H] + =392.2 7.

[0257]

[0258] In a single-necked flask, (2E)-9,10-dimethoxy-2-[(2,4,6-trimethylphenyl)imino]-3H,6H,7H-pyrimido[4,3-a]isoquinolin-4-one (2.0 g, 5.1 mmol, 1.0 equiv), ethyl 4-bromovalerate (2.1 g, 10.2 mmol, 2.0 equiv), 1,4-dioxane (100 mL), cesium carbonate (5.0 g, 15.3 mmol, 3.0 equiv), Xantphos (0.6 g, 1.0 mmol, 0.2 equiv), and Pd 2 (dba) 3 (0.5 g, 0.5 mmol, 0.1 equiv) were added successively. Under nitrogen protection, the reaction was carried out at 100 °C for 12 h. Normal-phase purification was carried out using DCM / MeOH (10 / 1) as the mobile phase to obtain ethyl 4-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)valerate (500 mg, 18.8% yield).

[0259] LCMS(ESI,m / z):[M+H] + =520.3 8.

[0261]

[0262] Ethyl 4-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)valerate (500 mg, 1.0 mmol, 1.0 equiv), methanol (50 mL), and an aqueous solution of sodium hydroxide (77 mg, 2.0 mmol, 2.0 equiv) in water (10 mL) were successively added to a single-necked flask, and the reaction was carried out at room temperature for 3 h. The solvent was evaporated, water (20 mL) was added, the pH was adjusted to 5 with dilute hydrochloric acid, and the mixture was extracted with EA (3 x 50 mL). The combined organic phases were dried over anhydrous sodium sulfate, and the organic phase was evaporated to obtain 4-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)valeric acid (450 mg, 95.1% yield).

[0263] LCMS(ESI,m / z):[M+H] + =492.2 9.

[0265]

[0266] 4-({9,10-Dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)valeric acid (300 mg, 0.6 mmol, 1.0 equiv), 1,4-dioxane (100 mL), triethylamine (182 mg, 1.8 mmol, 3.0 equiv), and DPPA (336 mg, 1.2 mmol, 2.0 equiv) were successively added to a three-necked flask. Under nitrogen protection, the reaction was carried out at room temperature for 1 h, then heated to 100 °C and reacted for 1 h. The temperature was cooled to 80 °C, and a solution of ammonia in tetrahydrofuran (1.4 M) (100 mL) was added. The reaction was carried out at room temperature for 2 h. The solvent was evaporated, and reverse-phase purification (column type: X-Bridge BEH C18 5 μm, 30 * 150 mm; mobile phase A: water (10 mmol / L NH 4 HCO 3 ), mobile phase B: acetonitrile; flow rate: 60 mL / min mL / min; gradient: 39% B to 48% B in 9 min; wavelength: 254 nm / 220 nm nm; RT1(min): 8) was carried out to obtain 3-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)butyrourea (94 mg, 30.46% yield).

[0267] LCMS(ESI,m / z):[M+H] + =506.3 10.

[0269]

[0270] The crude 3-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)butyrourea (50 mg) was resolved by Chiral-HPLC (Column type: CHIRAL ART Cellulose-SB, 2 * 25 cm, 5 μm; Mobile phase A: MtBE (0.1% DEA)-HPLC, Mobile phase B: MeOH:DCM = 1:1 - HPLC; Flow rate: 20 mL / min; Gradient: isocratic; Wavelength: 254 nm; RT1(min): 5.3; RT2(min): 7.2; Sample Solvent: MeOH:DCM = 1:1 - HPLC; Injection volume: 1 mL; Number Of Runs: 5) to obtain the product.

[0271] Compound 3-1 (isomer 1, RT1(min): 5.3, 17.6 mg, 34.78% yield)

[0272] LCMS (ESI, m / z): [M + H] + = 506.50

[0273] 1 H NMR (400 MHz, Methanol-d4) δ 7.16–7.08 (m, 2H), 6.93 (s, 1H), 6.64 (s, 1H), 5.41 (s, 1H), 4.59–4.55 (m, 1H), 4.19–4.02 (m, 2H), 3.89 (s, 3H), 3.66 (s, 3H), 3.38–3.34 (m, 1H), 3.25–3.21 (m, 1H), 2.97 (t, J = 6.4 Hz, 2H), 2.38 (s, 3H), 2.34–2.30 (m, 1H), 2.23 (s, 3H), 2.08 (s, 3H), 1.80–1.76 (m, 1H), 0.99 (d, J = 6.9 Hz, 3H).

[0274] Compound 3-2 (isomer 2, RT2(min): 7.2, 17.8 mg, 34.78% yield)

[0275] LCMS (ESI, m / z): [M + H] + = 506.50

[0276] 1 H NMR (400 MHz, Methanol-d 4)δ 7.16–7.08 (m, 2H), 6.93 (s, 1H), 6.64 (s, 1H), 5.41 (s, 1H), 4.59–4.45 (m, 1H), 4.19–4.02 (m, 2H), 3.89 (s, 3H), 3.66 (s, 3H), 3.38–3.34 (m, 1H), 3.25–3.21 (m, 1H), 2.97 (t, J = 6.4 Hz, 2H), 2.38 (s, 3H), 2.34–2.30 (m, 1H), 2.23 (s, 3H), 2.08 (s, 3H), 1.80–1.76 (m, 1H), 0.99 (d, J = 6.9 Hz, 3H).

[0277] Example 4, Compound 4-1 and Compound 4-2

[0278]

[0279] Preparation steps: 1.

[0281]

[0282] At room temperature and in an air environment, dissolve 2,4,6-trimethylaniline (3.0 g, 22.2 mmol, 1.0 equivalent) in MeOH (30 mL), and add NaBH 3 CN (1.0 g, 44.3 mmol, 2.0 equivalents) and acetic acid (1 mL, 17.4 mmol, 0.7 equivalent). The resulting mixture is stirred overnight under a nitrogen atmosphere at room temperature. The resulting mixture is concentrated under reduced pressure. The reactant is quenched with water at room temperature. The resulting mixture is extracted with ethyl acetate (2 x 100 mL). The combined organic phases are washed with saturated brine (3 x 200 mL) and dried over anhydrous sodium sulfate. The filtrate is concentrated under reduced pressure to obtain tert-butyl N-{3-[(2,4,6-trimethylphenyl)amino]cyclobutyl}carbamate (1.2 g, yield 16%).

[0283] LCMS (ESI): [M + H]+ = 305.1 2.

[0285]

[0286] tert-Butyl N-{3-[(2,4,6-trimethylphenyl)amino]cyclobutyl}carbamate (1.2 g, 3.9 mmol, 1.0 equiv) and 2-chloro-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (1.3 g, 4.7 mmol, 1.2 equiv) were added to isopropanol (15 ml) at room temperature. The reaction mixture was heated to 100 °C and stirred overnight under a nitrogen atmosphere at 100 °C. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether / ethyl acetate (2 / 1), to give 2-[(3-aminocyclobutyl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (150 mg, 6% yield).

[0287] LC-MS (ESI): [M+H] + = 461.2 3.

[0289]

[0290] At room temperature, N1-(2,4,6-trimethylphenyl)cyclobutane-1,3-diamine (150 mg, 0.7 mmol, 1.0 equiv) was dissolved in DCM (1 ml) in air, and solid phosgene (193 mg, 0.6 mmol, 2.0 equiv) was added to the solution. The resulting mixture was stirred for 1 h under a nitrogen atmosphere at room temperature. At room temperature, a methanol solution of ammonia (15 mL, 7 M) was added to the above mixture. The resulting mixture was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, acetonitrile water (10 mmol / L NH 4 HCO 3 ), 30%-47% gradient, 7 min; detector, UV 254 nm. The crude product was purified by preparative high-performance liquid chromatography under the following conditions (column: CHIRAL ARTCellulose-SB, 2*25 cm, 5 μm; mobile phase A: MtBE (0.1% DEA)-HPLC, mobile phase B: MeOH:DCM = 1:1-HPLC; flow rate: 20 mL / min; gradient: isocratic; wavelength: 220 nm; RT1 (min): 6.5; RT2 (min): 9.4; SampleSolvent: injection volume: 0.7 mL; number of runs: 3), to give the product.

[0291] Compound 4-1 (isomer 1, RT1 (min): 6.5, 16.3 mg, 9% yield)

[0292] LCMS(ESI): [M+H] + = 504.30

[0293] 1 H NMR(400MHz, Methanol-d 4 ) δ 7.20(s, 1H), 7.05(s, 2H), 6.93(s, 1H), 6.11(s, 1H), 4.56–4.47(m, 1H), 4.28–4.15(m, 1H), 4.03(t, J = 6.2Hz, 2H), 3.90–3.88(m, 6H), 2.93(t, J = 6.2Hz, 2H), 2.78–2.66(m, 2H), 2.32(s, 3H), 2.23(s, 6H), 1.88–1.73(m, 2H).

[0294] Compound 4-2 (isomer 2, RT2 (min): 9.4, 15.5 mg, 10% yield)

[0295] LCMS(ESI): [M+H] + = 504.40

[0296] 1 H NMR(400MHz, Methanol-d 4 ) δ 7.26(s, 1H), 7.05(s, 2H), 6.95(s, 1H), 6.25(s, 1H), 5.05–4.90(m, 1H), 4.28–4.15(m, 1H), 4.04(t, J = 6.4Hz, 2H), 3.92(s, 6H), 2.95(t, J = 6.4Hz, 2H), 2.34–2.32(m, 7H), 2.22(s, 6H).

[0297] Example 5, Compound 5-1 and Compound 5-2

[0298]

[0299] Preparation steps: 1.

[0301]

[0302] 2-Chloro-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (200 mg, 0.7 mmol, 1.0 equiv) and 2,4,6-trimethylaniline (369 mg, 2.7 mmol, 4.0 equiv) were dissolved in isopropanol (4 mL), and the mixture was stirred at 90 °C under nitrogen protection for 3 h. The resulting mixture was concentrated under reduced pressure. Purification by column chromatography using DCM / MeOH (10 / 1) gave (2E)-9,10-dimethoxy-2-[(2,4,6-trimethylphenyl)imino]-3H,6H,7H-pyrimido[4,3-a]isoquinolin-4-one (100 mg, yield 37.39%).

[0303] LCMS(ESI,m / z):[M+H] + =392.1 2.

[0305]

[0306] tert-Butyl 2-bromopropylcarbamate (304 mg, 1.3 mmol, 1.0 equiv) and Cs 2 CO 3 (1.0 g, 3.9 mmol, 3.0 equiv.) were added to a mixture solution of (2E)-9,10-dimethoxy-2-[(2,4,6-trimethylphenyl)imino]-3H,6H,7H-pyrimido[4,3-a]isoquinolin-1 (500 mg, 1.3 mmol, 1.0 equiv.) and dioxane (10 mL). Under nitrogen protection, Pd 2 (dba) 3 (117 mg, 0.1 mmol, 0.1 equiv.) and Xantphos (74 mg, 0.1 mmol, 0.1 equiv.) were added. The resulting mixture was stirred at 80 °C under nitrogen protection for 6 h. The resulting mixture was concentrated under reduced pressure. Purification by reverse-phase column chromatography was carried out under the following conditions: the column was C18 silica gel; the mobile phase was water, MeCN (0.1% FA), gradient 30% - 70%, 15 min; UV, 254 nm. tert-Butyl N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]carbamate (400 mg, yield 57.08%) was obtained.

[0307] LCMS(ESI,m / z):[M+H] + =549.6 3.

[0309]

[0310] Add N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]carbamate (100 mg, 0.2 mmol, 1.0 equiv) and a 1,4-dioxane (2 mL, 4 M) solution of HCl to a 25 mL round-bottom flask. The reaction mixture was stirred at 25 °C under nitrogen protection for 1 hour. The resulting mixture was concentrated under reduced pressure. 2-[(1-Aminoprop-2-en-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-1 (80 mg, crude product) was obtained.

[0311] LCMS(ESI, m / z): [M+H] + = 449.2 4.

[0313]

[0314] 2-[(1-Aminopropan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-1 (100 mg, 0.2 mmol, 1.0 equiv) and isocyano trimethylsilane (38 mg, 0.3 mmol, 1.5 equiv) were stirred in DCM (5 mL), and TEA (60 mg, 0.6 mmol, 3.0 equiv) was added dropwise under nitrogen protection at 25 °C. The reaction mixture was stirred at 25 °C under nitrogen protection for 2 hours. The resulting mixture was concentrated under reduced pressure. Purification by column chromatography was performed using a C18 silica gel column; mobile phase: ACN aqueous solution (0.1% NH 3 .H 2 O), gradient 25% - 75%, 20 min; UV, 254 nm. 2-({9,10-Dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propylurea (80 mg, yield 73.0%) was obtained.

[0315] LCMS(ESI, m / z): [M+H] + = 492.2 5.

[0317]

[0318] 2-({9,10-Dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propylurea (100 mg, 0.2 mmol, 1.0 eq) and an aqueous solution of hydrobromic acid (5 mL) were added to a 50 mL round-bottom flask. The resulting mixture was stirred for 3 hours under nitrogen protection at 120 °C. The mixture was extracted with ethyl acetate (2 × 50 mL), and the organic layer was washed with saturated brine (3 × 100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. 2-({9,10-Dihydroxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propylurea (80 mg, crude product) was obtained.

[0319] LCMS(ESI,m / z):[M+H] + = 464.2 6.

[0321]

[0322] 2-({9,10-Dihydroxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propylurea (100 mg, 0.2 mmol, 1.0 eq) and K 2 CO 3 (149 mg, 1.0 mmol, 5.0 eq) were added to DMF (1.5 mL), and ethyl 2-bromo-2,2-difluoroacetate (44 mg, 0.2 mmol, 1.0 eq) was added dropwise at room temperature. The reaction mixture was stirred for 1.5 hours under nitrogen protection at 55 °C. The resulting mixture was concentrated under reduced pressure. Purification by reverse-phase column chromatography was carried out under the following conditions: the chromatographic column was C18 silica gel; the mobile phase was water, ACN (0.1% FA), with a gradient of 20% to 70% for 20 min; UV detection was at 254 nm. 2-{[9-(Difluoromethoxy)-10-hydroxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl](2,4,6-trimethylphenyl)amino}propylurea (10 mg, yield 9.03%) was obtained.

[0323] LCMS(ESI,m / z):[M+H] + = 514.2 7.

[0325]

[0326] 2-{[9-(Difluoromethoxy)-10-hydroxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl](2,4,6-trimethylphenyl)amino}propylurea (10 mg, 0.02 mmol, 1.0 eq) and K2 CO 3 (4 mg, 0.03 mmol, 1.5 equiv), the solvent DMF (0.5 mL), methyl iodide (4 mg, 0.03 mmol, 1.5 equiv) was added dropwise at room temperature. The resulting mixture was stirred under nitrogen protection at 60 °C for 1.5 h. The obtained mixture was concentrated under reduced pressure. Purification by high performance liquid chromatography, column type: XBridge BEH C18 5 μm, 30 * 150 mm; mobile phase A: water (10 mmol / L NH 4 HCO 3 ), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 40% B to 50% B, 8 min; wavelength: 254 nm / 220 nm nm; RT1 (min): 7.3 to obtain the racemic product. The racemic product was purified by chiral high performance liquid chromatography (column: chiral ART Cellulose-SB, 2 * 25 cm, 5 μm; mobile phase A: MeOH:DCM = 1:1 - HPLC, mobile phase B: MtBE (0.1% DEA) - HPLC; flow rate: 20 ml / min; gradient: isocratic 50; wavelength: 220 nm; RT1 (min): 4.4; RT2 (min): 10.7; sample solvent: MeOH - HPLC; injection volume: 1 ml) to obtain the product.

[0327] Compound 5-1 (isomer 1, RT1 (min): 4.4, 1.54 mg, yield 14.87%).

[0328] LCMS (ESI, m / z): [M + H] + = 529.40.

[0329] 1 1H NMR (400 MHz, Methanol-d4) δ 7.20–7.05 (m, 3H), 6.95–6.82 (m, 1H), 6.80 (s, 1H), 5.49 (s, 1H), 4.49 (s, 1H), 4.16–4.10 (m, 2H), 3.72 (s, 3H), 2.97 (t, J = 6.4 Hz, 2H), 2.37 (s, 3H), 2.23 (s, 3H), 2.10 (s, 3H), 1.35–1.28 (m, 2H), 1.06 (s, 3H).

[0330] Compound 5-2 (isomer 2, RT2 (min): 10.7, 2.19 mg, yield 20.9%)

[0331] LCMS (ESI, m / z): [M + H] + = 529.45

[0332] 1 1H NMR (400 MHz, Methanol-d4) δ 7.20–7.05 (m, 3H), 6.95–6.82 (m, 1H), 6.80 (s, 1H), 5.49 (s, 1H), 4.49 (s, 1H), 4.16–4.10 (m, 2H), 3.72 (s, 3H), 2.96 (t, J = 6.4 Hz, 2H), 2.37 (s, 3H), 2.22 (s, 3H), 2.11 (s, 3H), 1.35–1.28 (m, 2H), 1.06 (s, 3H).

[0333] Example 6, Compound 6

[0334]

[0335] Preparation steps: 1.

[0337]

[0338] Dissolve trimethylaniline (1.0 g, 7.4 mmol, 1.0 equiv), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (3.4 g, 8.8 mmol, 1.2 equiv) and N,N-diisopropylethylamine (3.9 mL, 22.2 mmol, 3.0 equiv) in 20 mL of dichloromethane and stir at room temperature for 2 h. Concentrate under reduced pressure, dissolve in 10 mL of water, extract with ethyl acetate (3 × 10 mL), wash the organic layer with 20 mL of saturated sodium chloride, dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify the crude product by normal-phase column chromatography under the following conditions (mobile phase A: petroleum ether, mobile phase B: ethyl acetate; flow rate: 60 mL / min; gradient: 10% B to 50% B in 20 min; wavelength: 254 nm;) to obtain 2-(1,3-dioxoisoindolin-2-yl)-N-methylsulfonylacetamide (600 mg, yield 25.17%).

[0339] LCMS (ESI): [M+H] + = 323.14 2.

[0341]

[0342] 2-(1,3-dioxoisoindol-2-yl)-N-(2,4,6-trimethylphenyl)acetamide (600 mg, 1.8 mmol, 1.0 equiv), 2-chloro-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (653 mg, 2.2 mmol, 1.2 equiv), diphenyl O,O-(1-acetoxy-2,2,2-trichloroethyl)phosphonate (118.40 mg, 0.186 mmol, 0.1 equiv), [4-(N,N-dimethylamino)phenyl](di-tert-butylphosphino)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (49.39 mg, 0.186 mmol, 0.1 equiv) and cesium carbonate (1.5 g, 4.6 mmol, 2.5 equiv) were dissolved in 10 mL of 1,4-dioxane, protected by nitrogen, and stirred at 110 °C overnight. Concentrated under reduced pressure, dissolved in 10 mL of water, extracted with ethyl acetate (3×20 mL), the organic layer was washed with 20 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse-phase column chromatography under the following conditions (column: C18; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 30% B to 50% B in 20 min; wavelength: 254 nm;) to obtain 2-amino-N-(9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)-N-methylsulfonylacetamide (50 mg, yield 5.04%).

[0343] LCMS(ESI): [M+H] + = 449.16 3.

[0345]

[0346] Dissolve 2-amino-N-(9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)-N-methylsulfonylacetamide (20 mg, 0.05 mmol, 1.0 equiv), triethylamine (14 mg, 0.15 mmol, 3.0 equiv) and N,N'-carbonyldiimidazole (10.9 mg, 0.07 mmol, 1.5 equiv) in 2 mL of dichloromethane, protect with nitrogen, and add dropwise a 1.4-dioxane solution of 2 mL of amine at 0 °C. Stir at room temperature for 2 hours. Purify the crude product by reverse-phase column chromatography under the following conditions (column: SunFire C18 5 μm, 30 mm X 150 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 45% B to 59% B in 7 minutes; wavelength: 254 nm / 220 nm; time: 5.92) to obtain N-(9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)-N-methylsulfonyl-2-ureidoacetamide (2.9 mg, yield 13.10%).

[0347] LCMS(ESI, m / z): [M+H] + = 475.20

[0348] 1 1H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 7.71 (s, 1H), 7.18 (s, 1H), 7.08 - 7.01 (m, 3H), 4.77 (s, 2H), 4.08 (t, J = 6.6 Hz, 2H), 3.86 (s, 3H), 3.79 (s, 3H), 2.97 (t, J = 6.6 Hz, 2H), 2.30 (s, 3H), 2.12–2.10 (s, 6H).

[0349] Example 7, Compound 7-1, Compound 7-2 and Compound 7-3

[0350]

[0351] Preparation steps: 1.

[0353]

[0354] To methyl 2-bromocyclopropane-1-carboxylate (200 mg, 1.1 mmol, 1.0 equiv) and (2E)-9,10-dimethoxy-2-[(2,4,6-trimethylphenyl)imino]-3H,6H,7H-pyrimido[4,3-a]isoquinolin-4-one (437 mg, 1.1 mmol, 1.0 equiv) in 1,4-dioxane (80 mL) was added cesium carbonate (1.1 g, 3.4 mmol, 3.0 equiv), Xantphos (106 mg, 0.2 mmol, 0.2 equiv) and Pd 2 (dba) 3 (64 mg, 0.1 mmol, 0.1 equiv). After stirring at 100 °C for 12 h under a nitrogen atmosphere, the resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with methanol / dichloromethane (1 / 10) to give the crude product. The crude product was chromatographed (YMC Triart C18 ExRs 5 μm, 30 mm * 150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 39% B to 62% B over 10 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 8.33) to give methyl 2-[(2E)-9,10-dimethoxy-4-oxo-2-[(2,4,6-trimethylphenyl)imino]-6H,7H-pyrimido[4,3-a]isoquinolin-3-yl]cyclopropane-1-carboxylate (100 mg, yield 18.3%).

[0355] LCMS (ESI): [M+H] + = 490.2 2.

[0357]

[0358] At room temperature, sodium hydroxide (65 mg, 1.6 mmol, 5.0 equiv) in water (5 mL) was added to methyl 2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)cyclopropane-1-carboxylate (160 mg, 0.3 mmol, 1.0 equiv) in methanol (20 mL). The above mixture was stirred at 50 °C for 18 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The resulting mixture was extracted with ethyl acetate (3 x 50 mL). The aqueous layer was acidified to pH = 4 with 6 M aqueous hydrochloric acid. The resulting mixture was extracted with dichloromethane (3 x 100 mL). The combined organic layers were washed with brine and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give 2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)cyclopropane-1-carboxylic acid (170 mg, crude).

[0359] LCMS(ESI):[M+H] + =476.2 3.

[0361]

[0362] Oxalyl chloride (0.1 mL, 1.1 mmol, 3.0 equiv) was added dropwise to 2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)cyclopropane-1-carboxylic acid (170 mg, 0.4 mmol, 1.0 equiv) in dichloromethane (10 mL). The resulting mixture was stirred at 0 °C for 1 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The crude product was used directly without further purification. The crude product was dissolved in dichloromethane (1 mL) and added to a solution of ammonia in tetrahydrofuran (60 mL, 1.3 M) at 0 °C. The resulting mixture was stirred at 0 °C for 1 h. The resulting mixture was extracted with ethyl acetate (3 x 60 mL). The combined organic layers were washed with brine and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give 2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)cyclopropane-1-carboxamide (180 mg, crude).

[0363] LCMS(ESI):[M+H] + =475.2 4.

[0365]

[0366] A mixture of 2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)cyclopropane-1-carboxamide (180 mg, 0.4 mmol, 1.0 equiv) in tetrahydrofuran (20 mL) was cooled to 0 °C, and sodium borohydride (143 mg, 3.8 mmol, 10.0 equiv) was added portionwise. Iodine grains (963 mg, 3.8 mmol, 10.0 equiv) were dissolved in tetrahydrofuran (3 mL) and added to the resulting mixture. The resulting mixture was stirred at 70 °C for 18 h. The reaction was quenched by the addition of water (30 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (2 x 50 mL). The combined organic layers were washed with brine and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (dichloromethane / methanol = 10 / 1) to give 2-{[2-(aminomethyl)cyclopropyl](2,4,6-trimethylphenyl)amino}-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (45 mg, yield 25.8%).

[0367] LCMS(ESI):[M+H] + = 461.2 5.

[0369]

[0370] A mixture of 2-{[2-(aminomethyl)cyclopropyl](2,4,6-trimethylphenyl)amino}-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (45 mg, 0.1 mmol, 1.0 equiv), triethylamine (30 mg, 0.3 mmol, 3.0 equiv) and trimethylsilyl isocyanate (23 mg, 0.2 mmol, 2.0 equiv) in DCM (5 mL) was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The crude product (45 mg) was purified by Prep-HPLC under the following conditions (column: XBridge BEH C18 OBD Prep Column 130, 5 μm, 19 mm * 250 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate); flow rate: 25 mL / min; gradient: 45% B to 67% B for 10 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 7.93) to give [2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)cyclopropyl]methylurea (20 mg, yield 40.65%).

[0371] LCMS(ESI):[M+H]+ = 504.3 6.

[0373]

[0374] Compound 7-1, isomer 1 (Column: CHIRAL ART Amylose-SA, 2 * 25 cm, 5 μm; Mobile phase A: methanol:dichloromethane = 1:1, Mobile phase B: methyl tert-butyl ether (0.1% diethylamine) Flow rate: 20 mL / min; Gradient: isocratic; Wavelength: 254 nm; RT1 (min): 6.7; Sample solvent: methanol (0.1% diethylamine); Injection volume: 1 mL; Number of runs: 3) gave the product (3.8 mg, yield 12.43%).

[0375] LCMS (ESI): [M + H] + = 504.30

[0376] 1 H NMR (400 MHz, Methanol-d4) δ 7.10 (s, 1H), 7.07 (s, 1H), 6.95 (s, 1H), 6.72 (s, 1H), 5.54 (s, 1H), 4.25–4.06 (m, 2H), 3.90 (s, 4H), 3.68 (s, 3H), 2.99 (t, J = 6.6 Hz, 3H), 2.48–2.40 (m, 1H), 2.36 (s, 3H), 2.23 (s, 3H), 2.03 (s, 3H), 1.43–1.31 (m, 2H).

[0377] Compound 7-2, isomer 2 (Column: CHIRAL ART Amylose-SA, 2 * 25 cm, 5 μm; Mobile phase A: methanol:dichloromethane = 1:1, Mobile phase B: methyl tert-butyl ether (0.1% diethylamine) Flow rate: 20 mL / min; Gradient: isocratic; Wavelength: 254 nm; RT2 (min): 10.9; Sample solvent: methanol (0.1% diethylamine); Injection volume: 1 mL; Number of runs: 3) gave the product (3.5 mg, yield 11.29%).

[0378] LCMS (ESI): [M + H] + = 504.35

[0379] 11H NMR (400 MHz, Methanol-d4) δ 7.10 (s, 1H), 7.07 (s, 1H), 6.95 (s, 1H), 6.72 (s, 1H), 5.54 (s, 1H), 4.25–4.06 (m, 2H), 3.90 (s, 4H), 3.68 (s, 3H), 2.99 (t, J = 6.6 Hz, 3H), 2.48–2.40 (m, 1H), 2.36 (s, 3H), 2.23 (s, 3H), 2.03 (s, 3H), 1.43–1.31 (m, 2H).

[0380] Compound 7-3, isomer 3 (Column: XBridge BEH C18 OBD Prep Column 130, 5 μm, 19 mm * 250 mm; Mobile phase A: water (10 mmol / L ammonium bicarbonate); Mobile phase B: acetonitrile; Flow rate: 60 mL / min; Gradient: 39% B to 61% B for 9 minutes; Wavelength: 254 nm / 220 nm; RT1 (min): 5.45), the product was obtained (20 mg, yield 40.6%).

[0381] LCMS (ESI): [M + H] + = 504.35

[0382] 1 1H NMR (400 MHz, Methanol-d4) δ 7.10 (s, 1H), 7.07 (s, 1H), 7.02–6.89 (m, 2H), 6.70 (s, 1H), 5.57 (s, 1H), 4.61 (s, 6H), 4.27–4.04 (m, 2H), 4.05–3.91 (m, 2H), 3.90 (s, 3H), 3.85–3.72 (m, 1H), 3.68 (s, 3H), 3.05–2.95 (m, 2H), 2.92–2.87 (m, 1H), 2.36 (s, 3H), 2.31–2.14 (m, 8H).

[0383] Example 8, Compound 8

[0384]

[0385] Preparation steps 1.

[0387]

[0388] tert-Butyl N-(2-hydroxypropyl)carbamate (15.0 g, 85.6 mmol, 1.0 equiv) and TEA (26.0 g, 256.8 mmol, 1.5 equiv) were stirred in DCM (80 mL), and then DMAP (1.1 g, 8.6 mmol, 0.1 equiv) and TsCl (21.2 g, 111.3 mmol, 1.3 equiv) were added portionwise at 0 °C. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was extracted with dichloromethane (3 x 200 mL). The organic layer was washed with brine and dried over anhydrous sodium sulfate. The filtered filtrate was concentrated under reduced pressure and purified by silica gel column chromatography, eluting with PE / EA (5 / 1) to give tert-Butyl N-[2-[(4-methylphenylsulfonyl)oxy]propyl]carbamate (20.3 g, yield 71.9%).

[0389] LC-MS (ESI): [M+H] + = 330.1 2.

[0391]

[0392] To a solution of 9,10-dimethoxy-2-[(2,4,6-trimethylphenyl)amino]-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (1.6 g, 4.1 mmol, 1.0 equiv) in 1,4-dioxane (20 mL) was added tert-Butyl N-[2-[(4-methylphenylsulfonyl)oxy]propyl]carbamate (2.0 g, 6.1 mmol, 1.5 equiv), cesium carbonate (2.7 g, 8.2 mmol, 2.0 equiv), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.5 g, 0.8 mmol, 0.2 equiv) and tris(dibenzylideneacetone)dipalladium (0.4 g, 0.4 mmol, 0.1 equiv). The mixture was purged with nitrogen and then heated to 100 °C. Under nitrogen, the mixture was stirred at 100 °C for 5 h. The reaction system was extracted with ethyl acetate (3 x 200 mL) and water (150 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and rotary evaporated. The resulting residue was purified by reverse phase column chromatography with the following conditions: Column: XBridge BEH C18, 5 μm, 30*150 mm; Mobile phase A: water (10 mmol / L ammonium bicarbonate), Mobile phase B: acetonitrile; Flow rate: 60 mL / min; Gradient: 55% B to 65% B in 7 min; Wavelength: 254 nm / 220 nm; RT1 (min): 5.67 / 6.05. To give tert-Butyl N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]carbamate (35.33 mg, yield 1.55%).

[0393] LC-MS (ESI): [M+H] + = 549.35

[0394] 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.11 (d, J = 10.2 Hz, 2H), 6.93 (s, 1H), 6.64 (s, 1H), 5.41 (s, 1H), 4.57–4.39 (m, 1H), 4.18–4.02 (m, 2H), 3.89 (s, 3H), 3.66 (s, 3H), 3.55–3.48 (m, 1H), 3.40 (t, J = 8.2 Hz, 1H), 2.97 (t, J = 6.5 Hz, 2H), 2.37 (s, 3H), 2.22 (s, 3H), 2.11 (s, 3H), 1.46 (s, 9H), 1.31 (s, 1H), 1.08 (d, J = 6.9 Hz, 3H).

[0395] Example 9, Compound 9-1 and Compound 9-2

[0396]

[0397] Preparation steps: 1.

[0399]

[0400] 2-((1-Aminopropan-2-yl)(methylsulfonyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (150 mg, 0.33 mmol, 1.0 equiv), 3H-1,2,3-triazole-4-carboxylic acid (56 mg, 0.5 mmol, 1.5 equiv), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (96 mg, 0.5 mmol, 1.5 equiv), 1-hydroxybenzotriazole (67 mg, 0.5 mmol, 1.5 equiv) were dissolved in 2 ml of DMF, protected by nitrogen, stirred at room temperature for 1 hour, and concentrated under reduced pressure. The crude product was purified by reverse-phase column chromatography under the following conditions (column: Xselect CSH C18 5 μm, 30 mm X 150 mm; mobile phase A: water (0.1% FA), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 27% B to 55% B in 9 min; wavelength: 254 nm / 220 nm; RT1 (min): 6.77 / 8.43) to obtain N-(2-((9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)(methylsulfonamido)propyl)-1H-1,2,3-triazole-5-carboxamide (30 mg, yield 16.5%).

[0401] LCMS (ESI): [M+H] + = 544.25 2.

[0403]

[0404] The racemate (30 mg) was purified by Chiral-SFC under the following conditions (column: CHIRALPAK IH 3*25 cm, 5 μm; mobile phase A: CO 2 , mobile phase B: methanol (0.1% 2M NH 3 -MEOH); flow rate: 85 mL / min; gradient: isocratic 35% B; column temperature (°C): 35; pressure (bar): 100; wavelength: 220 nm; RT1 (min): 2.92; RT2 (min): 4.42; sample solvent: DCM (0.1% 2M NH 3 -MeOH)--HPLC; injection volume: 1 mL) to obtain the product

[0405] Compound 9-1 (isomer 1, RT1 (min): 2.92, 9.2 mg, yield 28.58%)

[0406] LCMS (ESI): [M+H] + = 544.25

[0407] 1 1H NMR (400 MHz, Methanol-d 4 ) δ 8.25 (s, 1H), 7.15–7.11 (m, 2H), 6.99–6.92 (m, 1H), 6.65 (s, 1H), 5.44 (s, 1H), 4.74 (s, 1H), 4.23–4.04 (m, 2H), 3.95 - 3.88 (m, 4H), 3.85–3.75 (m, 1H), 3.67 (s, 3H), 2.99 (t, J=6.4 Hz, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 2.13 (s, 3H), 1.31 (d, J=4.0 Hz, 1H), 1.11 (d, J=6.8 Hz, 3H).

[0408] Compound 9-2 (isomer 2, RT2 (min): 4.42, 8.8 mg, yield 24.5%)

[0409] LCMS (ESI): [M+H] + = 544.25

[0410] 1 1H NMR (400 MHz, Methanol-d 4 ) δ 8.25 (s, 1H), 7.15–7.11 (m, 2H), 6.99–6.92 (m, 1H), 6.65 (s, 1H), 5.44 (s, 1H), 4.74 (s, 1H), 4.23–4.04 (m, 2H), 3.95 - 3.88 (m, 4H), 3.85–3.75 (m, 1H), 3.67 (s, 3H), 2.99 (t, J=6.4 Hz, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 2.13 (s, 3H), 1.31 (d, J=4.0 Hz, 1H), 1.11 (d, J=6.8 Hz, 3H).

[0411] Example 10, Compound 10-1 and Compound 10-2

[0412]

[0413] Preparation steps: 1.

[0415]

[0416] 2-((1-Aminopropan-2-yl)(methylsulfonyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (150 mg, 0.33 mmol, 1.0 equiv), imidazole-2-carboxylic acid (56 mg, 0.5 mmol, 1.5 equiv), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (96 mg, 0.5 mmol, 1.5 equiv), 1-hydroxybenzotriazole (67 mg, 0.5 mmol, 1.5 equiv) were dissolved in 2 ml of DMF, protected by nitrogen, stirred at room temperature for 1 hour, and concentrated under reduced pressure. The crude product was purified by reverse-phase column chromatography under the following conditions (column: Xselect CSH C18 5 μm, 30 mm X 150 mm; mobile phase A: water (0.1% FA), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 27% B to 55% B in 9 minutes; wavelength: 254 nm / 220 nm; RT1(min): 6.77 / 8.43) to obtain N-(2-((9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)(methylsulfonamido)propyl)-1H-imidazole-2-carboxamide (70 mg, yield 44.0%).

[0417] LCMS(ESI): [M+H] + = 544.25 2.

[0419]

[0420] The racemate (70 mg) was purified by Chiral-SFC under the following conditions (column: CHIRAL ART Cellulose-SB, 3*25 cm, 5 μm; mobile phase A: CO 2 , mobile phase B: MeOH (0.1% 2M NH 3 -MeOH); flow rate: 100 mL / min; gradient: isocratic 25% B; column temperature (°C): 35; pressure (bar): 100; wavelength: 220 nm; RT1(min): 9.4; RT2(min): 10.72; sample solvent: MeOH (0.1% DEA)--HPLC; injection volume: 1 mL) to obtain the product

[0421] Compound 10-1 (isomer 1, RT1(min): 9.4, 24.9 mg, yield 34.5%).

[0422] LCMS(ESI): [M+H] + = 543.25

[0423] 11H NMR (400 MHz, Methanol-d 4 ) δ 7.21 (s, 3H), 7.16–7.08 (m, 2H), 6.94 (s, 1H), 6.65 (s, 1H), 5.43 (s, 1H), 4.70 (s, 1H), 4.21–4.02 (m, 2H), 3.97–3.91 (m, 1H), 3.89 (s, 4H), 3.82–3.84 (m, 1H), 3.67 (s, 3H), 3.05–2.94 (m, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 2.14 (s, 3H), 1.31 (s, 1H), 1.15 (d, J = 6.8 Hz, 3H).

[0424] Compound 10-2 (isomer 2, RT2 (min): 10.72, 24.4 mg, yield 34.5%)

[0425] LCMS (ESI): [M+H] + = 543.25

[0426] 1 1H NMR (400 MHz, Methanol-d 4 ) δ 7.21 (s, 3H), 7.16–7.08 (m, 2H), 6.94 (s, 1H), 6.65 (s, 1H), 5.43 (s, 1H), 4.70 (s, 1H), 4.21–4.02 (m, 2H), 3.97–3.91 (m, 1H), 3.89 (s, 4H), 3.84–3.76 (m, 1H), 3.67 (s, 3H), 3.05–2.94 (m, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 2.14 (s, 3H), 1.31 (s, 1H), 1.15 (d, J = 6.8 Hz, 3H).

[0427] Example 11, Compound 11-1 and Compound 11-2

[0428]

[0429] Preparation steps: 1.

[0431]

[0432] 2H-Pyrazole-3-carboxylic acid (25 mg, 0.2 mmol, 1.0 equiv) was dissolved in SOCl 2(1 mL), stirred at room temperature for 2 h under nitrogen protection. The resulting mixture was concentrated to dryness by rotary evaporation under vacuum. The above mixture was dissolved in DCM (1 mL) and added dropwise to a solution of 2-[(1-aminopropan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (100 mg, 0.2 mmol, 1.0 equiv) and N,N-diisopropylethylamine (86 mg, 0.7 mmol, 3.0 equiv) in DCM (5 mL) at 0 °C under nitrogen protection. The reaction mixture was stirred at room temperature for 2 h under nitrogen protection. The reaction mixture was quenched with water (5 mL) at room temperature, extracted with ethyl acetate (3 x 10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification was carried out by reverse phase, conditions: column was C18 silica gel; mobile phase, ACN (0.1% FA) in water, 10% - 50% gradient, 10 min; UV detector, 254 nm. N-[2-({9,10-Dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]-2H-pyrazole-3-carboxamide (40 mg, yield 32.0%) was obtained. 2.

[0434]

[0435] The crude product (40 mg) was purified by column chromatography: column: CHIRALPAK IH 3*25 cm, 5 μm; mobile phase A: CO 2 , mobile phase B: MEOH (0.1% DEA); flow rate: 80 mL / min; gradient: isocratic 40% B; column temperature (°C): 10 °C; head temperature (°C): 10 °C: 80 mL / min; gradient: 40% B isocratic; column temperature (°C): 35; pressure (bar): 100; wavelength: 220 nm; RT1 (min): 2.73; RT2 (min): 4.17; sample solvent: injection volume: 4.8 mL, and the product was obtained.

[0436] Compound 11-1 (isomer 1, RT1 (min): 2.73, 12.1 mg, yield 9.6%)

[0437] LCMS (ESI): [M+H] + = 543.55

[0438] 1 1H NMR (400 MHz, Methanol-d 4)δ 7.71 (d, J = 2.4 Hz, 1H), 7.13 (d, J = 13.6 Hz, 2H), 6.93 (s, 1H), 6.78 (d, J = 2.4 Hz, 1H), 6.65 (s, 1H), 5.43 (s, 1H), 4.70 (s, 1H), 4.16–4.10 (m, 2H), 3.91–3.78 (m, 5H), 3.67 (s, 3H), 2.98 (t, J = 6.4 Hz, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 2.13 (s, 3H), 1.11 (d, J = 6.8 Hz, 3H).

[0439] Compound 11-2 (isomer 2, RT2 (min): 4.17, 10.9 mg, yield 26.6%)

[0440] LCMS (ESI): [M+H] + = 543.35

[0441] 1 1H NMR (400 MHz, Methanol-d 4 )δ 7.71 (d, J = 2.4 Hz, 1H), 7.18–7.12 (m, 2H), 6.93 (s, 1H), 6.78 (d, J = 2.4 Hz, 1H), 6.65 (s, 1H), 5.43 (s, 1H), 4.75–4.55 (m, 1H), 4.16–4.10 (m, 2H), 3.91–3.78 (m, 5H), 3.67 (s, 3H), 2.98 (t, J = 6.4 Hz, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 2.13 (s, 3H), 1.11 (d, J = 6.8 Hz, 3H).

[0442] Example 12, Compound 12-1 and Compound 12-2

[0443]

[0444] Preparation steps: 1.

[0446]

[0447] In a high-pressure reactor, palladium on carbon (10%, 2.90 g) was added to ethyl 1-benzyl-5-methyl-1,2,3-triazole-4-carboxylate (850 mg, 3.5 mmol, 1.0 equiv) in 150 mL of methanol. The mixture was stirred at room temperature under a hydrogen pressure of 20 bar for 18 hours, filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (1 / 1), to give ethyl 5-methyl-1H-1,2,3-triazole-4-carboxylate (260 mg, yield 48.4%).

[0448] LCMS(ESI,m / z):[M+H] + =156.1 2.

[0450]

[0451] A mixture of ethyl 5-methyl-1H-1,2,3-triazole-4-carboxylate (260 mg, 1.7 mmol, 1.0 equiv) and sodium hydroxide (201 mg, 5.1 mmol, 3.0 equiv) in methanol (10 mL) and water (10 mL) was stirred at 60 °C for 18 hours. The resulting mixture was concentrated under reduced pressure. The mixture was acidified to pH = 5 with 2 M aqueous hydrochloric acid. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase chromatography under the following conditions: column, C18 silica gel; mobile phase, acetonitrile, water, 0%-10% gradient, 10 minutes; detector, UV 254 nm, to give 5-methyl-3H-1,2,3-triazole-4-carboxylic acid (200 mg, yield 93.9%).

[0452] LCMS(ESI):[M+H] + =128.0 3.

[0454]

[0455] At room temperature, 2-[(1-aminopropan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (100 mg, 0.2 mmol, 1.0 equiv) and 5-methyl-3H-1,2,3-triazole-4-carboxylic acid (28.3 mg, 0.2 mmol, 1.0 equiv) were dissolved in DMF (3 mL), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (64 mg, 0.3 mmol, 1.5 equiv) and 1-hydroxybenzotriazole (45 mg, 0.3 mmol, 1.5 equiv) were added. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase under the following conditions (column: XBridge BEH Shield RP18 5 μm, 30 mm * 150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: from 36% B to 54% B in 10 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 6.18), to obtain N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]-5-methyl-3H-1,2,3-triazole-4-carboxamide (30 mg, yield 24.1%).

[0456] LCMS(ESI):[M+H] + =558.3 4.

[0458]

[0459] The crude product (30 mg) was purified by Chiral-HPLC under the following conditions: CHIRALPAK IH 3 * 25 cm, 5 μm; mobile phase A: CO 2 , mobile phase B: methanol (0.1% diethylamine); flow rate: 85 mL / min; gradient: isocratic 35% B; column temperature (°C): 25; back pressure (bar): 100; wavelength: 254 nm; RT1 (min): 2.85; RT2 (min): 4.5 Sample solvent: methanol:dichloromethane = 1:1 - HPLC; injection volume: 1 mL) to obtain the product.

[0460] Compound 12-1 (isomer 1, RT1 (min): 2.85, 1.6 mg, yield 5.14%)

[0461] LCMS(ESI, m / z):[M+H] + =558.35

[0462] 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.17–7.10 (m, 2H), 6.93 (s, 1H), 6.65 (s, 1H), 5.43 (s, 1H), 4.78–4.69 (m, 1H), 4.20–4.04 (m, 2H), 3.97–3.77 (m, 5H), 3.67 (s, 3H), 2.98 (t, J = 6.5 Hz, 2H), 2.59–2.52 (m, 3H), 2.38 (s, 3H), 2.27 (s, 3H), 2.13 (s, 3H), 1.11 (d, J = 6.9 Hz, 3H).

[0463] Compound 12-2 (isomer 2, RT2 (min): 4.5, 3.2 mg, yield 9.86%)

[0464] LCMS (ESI, m / z): [M+H] + = 558.46

[0465] 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.17–7.10 (m, 2H), 6.93 (s, 1H), 6.65 (s, 1H), 5.43 (s, 1H), 4.78–4.69 (m, 1H), 4.20–4.04 (m, 2H), 3.97–3.77 (m, 5H), 3.67 (s, 3H), 2.98 (t, J = 6.5 Hz, 2H), 2.59–2.52 (m, 3H), 2.38 (s, 3H), 2.27 (s, 3H), 2.13 (s, 3H), 1.11 (d, J = 6.9 Hz, 3H).

[0466] Example 13, Compound 13-1 and Compound 13-2

[0467]

[0468] Preparation steps: 1.

[0470]

[0471] 2-[(1-Aminopropan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (100 mg, 0.2 mmol, 1.0 equiv), 3H-imidazole-4-carboxylic acid (25 mg, 0.2 mmol, 1.0 equiv), HOBT (45 mg, 0.3 mmol, 1.5 equiv), and EDCI (64 mg, 0.3 mmol, 1.5 equiv) were dissolved in DMF (5 mL) and stirred for 2 h under nitrogen protection at room temperature. The reaction solution was quenched with water (10 mL) at room temperature. The aqueous layer was extracted with ethyl acetate (3 x 20 mL), and the organic layer was washed with saturated brine (3 x 50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification was performed by reverse phase with the following conditions: column, C18 silica gel; mobile phase, acetonitrile-water (0.1% FA), gradient from 10% to 50% in 10 minutes; detector, UV 254 nm. N-[2-({9,10-Dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]-3H-imidazole-4-carboxamide (20 mg, yield 15.7%) was obtained after purification. 2.

[0473]

[0474] The crude product (20 mg) was purified by Chiral-HPLC: CHIRALPAK IH 3 * 25 cm, 5 μm; mobile phase A: CO 2 , mobile phase B: MeOH:DCM = 2:1 (0.1% 2M NH 3 -MeOH); flow rate: 100 mL / min; gradient: isocratic 18% B; column temperature (°C): 20 °C; column head temperature (°C): 20 °C: 100 mL / min; gradient: isocratic 18% B; column temperature (°C): 35; back pressure (bar): 100; wavelength: 220 nm; RT1 (min): 14.2; RT2 (min): 21.37; Sample Solvent: DCM--HPLC; injection volume: 2 mL, and the product was obtained.

[0475] Compound 13-1 (isomer 1, RT1 (min): 14.2, 2.04 mg, yield 1.6%)

[0476] LCMS (ESI): [M+H] + = 543.20

[0477] 1 1H NMR (400 MHz, Methanol-d 4)δ 7.75–7.70 (m, 2H), 7.15–7.11 (m, 2H), 6.98–6.92 (m, 1H), 6.65 (s, 1H), 5.43 (s, 1H), 4.75–4.55 (m, 1H), 4.15–4.09 (m, 2H), 3.90 (s, 3H), 3.85–3.80 (m, 2H), 3.67 (s, 3H), 2.98 (t, J = 6.4 Hz, 2H), 2.38 (s, 3H), 2.33–2.27 (m, 3H), 2.15 (s, 3H), 1.16 (d, J = 6.9 Hz, 3H).

[0478] Compound 13-2 (isomer 2, RT2 (min): 21.37, 2.72 mg, yield 13.0%)

[0479] LCMS (ESI): [M+H] + = 543.20

[0480] 1 1H NMR (400 MHz, Methanol-d 4 )δ 7.75–7.70 (m, 2H), 7.15–7.11 (m, 2H), 6.98–6.92 (m, 1H), 6.65 (s, 1H), 5.43 (s, 1H), 4.75–4.55 (m, 1H), 4.15–4.09 (m, 2H), 3.90 (s, 3H), 3.85–3.80 (m, 2H), 3.67 (s, 3H), 2.98 (t, J = 6.4 Hz, 2H), 2.38 (s, 3H), 2.33–2.27 (m, 3H), 2.15 (s, 3H), 1.16 (d, J = 6.9 Hz, 3H).

[0481] Example 14, Compound 14

[0482]

[0483] Preparation steps: 1.

[0485]

[0486] 2-[(1-Aminopropan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (250 mg, 0.6 mmol, 1.0 equiv), DCM (5 mL), DIEA (216 mg, 1.7 mmol, 3.0 equiv), cyanoacetic acid (57 mg, 0.7 mmol, 1.2 equiv) and HATU (318 mg, 0.8 mmol, 1.5 equiv) were successively added to a single-necked flask, and the reaction was carried out overnight at room temperature. The solvent was evaporated, and purification was performed by reverse phase (column type, C18 silica gel; mobile phase, acetonitrile-water (10 mmol / L NH 4 HCO 3 ), gradient 10% to 50% for 30 minutes; detection wavelength 254 nm) to obtain 2-cyano-N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]acetamide (200 mg, 69.6% yield).

[0487] LCMS (ESI, m / z): [M+H] + = 516.2 2.

[0489]

[0490] 2-Cyano-N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]acetamide (200 mg, 0.4 mmol, 1.0 equiv), 1,4-dioxane (30 mL), sodium acetate (95 mg, 1.2 mmol, 3.0 equiv) and acetic anhydride (48 mg, 0.5 mmol, 1.2 equiv) were successively added to a single-necked flask, and the reaction was carried out overnight at 100 °C. The solvent was evaporated, and purification was performed by reverse phase (column type: XBridge BEH Shield RP18 5 μm, 30 mm * 150 mm; mobile phase A: water (10 mmol / L NH 4 HCO 3 ), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 21% B to 44% B for 9 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 6.82) to obtain (2Z)-2-cyano-N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]-3-hydroxy-2-enamide (13.27 mg, 6.1% yield).

[0491] LCMS (ESI, m / z): [M+H] + = 558.25

[0492] 1 H NMR (400 MHz, Methanol-d4) δ 7.15–7.05 (m, 2H), 7.07–6.88 (m, 2H), 6.63 (s, 1H), 5.40 (s, 1H), 4.68–4.59 (m, 1H), 4.21–4.12 (m, 2H), 4.11–4.05 (m, 1H), 3.89 (s, 3H), 3.66 (s, 3H), 2.98 (t, J = 6.5 Hz, 2H), 2.37 (s, 3H), 2.33 (s, 1H), 2.25–2.23 (m, 6H), 2.14 (s, 3H), 1.09 (d, J = 6.7 Hz, 3H).

[0493] Example 15, Compound 15-1 and Compound 15-2

[0494]

[0495] Preparation steps: 1.

[0497]

[0498] Under an air environment at 0 °C, dissolve N-(2-hydroxybutyl)carbamic acid tert-butyl ester (500 mg, 2.6 mmol, 1.0 equivalent) and TEA (1.1 mL, 7.9 mmol, 3.0 equivalents) in DCM (5 mL) solution, and add TsCl (1.2 g, 3.9 mmol, 1.5 equivalents) and DMAP (32 mg, 0.3 mmol, 0.1 equivalent) respectively. The resulting mixture is stirred for 1.5 hours under a nitrogen environment at room temperature. The reaction is quenched with water at room temperature. The resulting mixture is extracted with dichloromethane (2 x 50 mL). The combined organic layers are washed with saturated brine (2 x 100 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate is concentrated under reduced pressure. The residue is purified by reverse phase with the following conditions: column, C18 silica gel; mobile phase, ACN water (10 mmol / L NH 4 HCO 3 ), gradient from 10% to 50% in 10 minutes; detector, UV 254 nm, to obtain N-{2-[(4-methylbenzenesulfonyl)oxy]butyl}carbamic acid tert-butyl ester (700 mg, yield 69%).

[0499] LCMS (ESI): [M+H] + = 361.2 2.

[0501]

[0502] 9,10 - Dimethoxy - 2 - [(2,4,6 - trimethylphenyl)amino] - 6H,7H - pyrimido[4,3 - a]isoquinolin - 4 - one (1.0 g, 2.5 mmol, 1.0 equiv), N - {2 - [(4 - methylbenzenesulfonyl)oxy]butyl}carbamic acid tert - butyl ester (1.3 g, 3.8 mmol, 1.5 equiv), and cesium carbonate (1.7 g, 5.108 mmol, 2.0 equiv) were added to N,N - dimethylformamide (10 mL). Under a room - temperature, air atmosphere, XantPhos (295 mg, 0.5 mmol, 0.2 equiv) and Pd 2 (dba) 3 (233 mg, 0.3 mmol, 0.1 equiv) were added. The resulting mixture was stirred overnight under a nitrogen atmosphere at 100 °C. The reaction was quenched with water at room temperature. The resulting mixture was extracted with ethyl acetate (2 x 50 mL). The combined organic layers were washed with saturated brine (2 x 100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse - phase chromatography under the following conditions: column, C18 silica gel; mobile phase, acetonitrile - water (10 mmol / L NH 4 HCO 3 ), 50% - 60% gradient, 10 min; detector, UV 254 nm. N - [2 - ({9,10 - dimethoxy - 4 - oxo - 6H,7H - pyrimido[4,3 - a]isoquinolin - 2 - yl}(2,4,6 - trimethylphenyl)amino)butyl]carbamic acid tert - butyl ester (330 mg, crude) was finally obtained.

[0503] LCMS(ESI): [M + H] + = 563.2 3.

[0505]

[0506] Under a nitrogen atmosphere at room temperature, tert-butyl N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)butyl]carbamate (330 mg, 0.6 mmol, 1.0 equiv) was dissolved in 1,4-dioxane (2 mL), and a 1,4-dioxane (2 mL, 4 M) solution of hydrochloric acid was added. The resulting mixture was stirred for 1.5 h under a nitrogen atmosphere at room temperature. The reaction was quenched with water at room temperature. The mixture was neutralized to pH = 9 with saturated sodium carbonate solution. The resulting mixture was extracted with ethyl acetate (2 x 50 mL). The combined organic layers were washed with saturated brine (2 x 100 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (DCM / MeOH = 20 / 1) to give 2-[(1-aminobutan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (150 mg, 49% yield).

[0507] LCMS(ESI):[M+H] + =463.2 4.

[0509]

[0510] 2-[(1-aminobutan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (100 mg, 0.2 mmol, 1 equiv) and triethylamine (65 mg, 0.6 mmol, 3.0 equiv) were dissolved in DCM (1 mL), and isocyano trimethylsilane (37 mg, 0.3 mmol, 1.5 equiv.) was added under air at 0 °C. The resulting mixture was stirred for 2 h under a nitrogen atmosphere at room temperature. The resulting mixture was concentrated under reduced pressure. The crude product (50 mg) was purified by preparative liquid chromatography, conditions (column: XBridge BEH C18 5, 19*250 mm; mobile phase A: water (10 mmol / L NH 4 HCO 3 )), mobile phase B: acetonitrile; flow rate: 25 mL / min; gradient: from 41% B to 48% B in 10 min; wavelength: 254 nm / 220 nm; RT(min): 7.25 / 8.32), to give the product.

[0511] Compound 15-1 (isomer 1, RT(min): 7.25, 4.02 mg, 4% yield)

[0512] LCMS(ESI):[M+H]+=506.25

[0513] 1 1H NMR (400 MHz, Methanol-d4) δ 7.15–7.06 (m, 2H), 6.93 (s, 1H), 6.63 (s, 1H), 5.41 (s, 1H), 4.21–4.02 (m, 3H), 3.89 (s, 3H), 3.66 (s, 4H), 2.97 (t, J = 8.0 Hz, 2H), 2.37 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H), 1.66–1.52 (m, 2H), 0.98 (t, J = 7.4 Hz, 3H).

[0514] Compound 15-2 (isomer 2, RT (min): 8.32, 4.02 mg, yield 4%)

[0515] LCMS (ESI): [M+H]+ = 506.25

[0516] 1 1H NMR (400 MHz, Methanol-d4) δ 7.15–7.06 (m, 2H), 6.93 (s, 1H), 6.63 (s, 1H), 5.41 (s, 1H), 4.21–4.02 (m, 3H), 3.89 (s, 3H), 3.66 (s, 4H), 2.97 (t, J = 8.0 Hz, 2H), 2.37 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H), 1.66–1.52 (m, 2H), 0.98 (t, J = 7.4 Hz, 3H).

[0517] Example 16, Compound 16

[0518]

[0519] Preparation steps 1.

[0521]

[0522] At room temperature, trimethylsilyl isocyanate (77 mg, 0.7 mmol, 1.5 equiv) was added to a solution of 2-[(1-aminopropan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (200 mg, 0.4 mmol, 1.0 equiv) and triethylamine (135 mg, 1.3 mmol, 3.0 equiv) in dichloromethane (5 mL). The resulting reaction mixture was stirred at room temperature for 2 h. The reaction mixture was extracted with dichloromethane (3 x 20 mL). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. After filtration of the resulting mixture, the filtrate was concentrated under reduced pressure to give 2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propylurea (200 mg, crude).

[0523] LCMS(ESI): [M+H] + = 492.3 2.

[0525]

[0526] At room temperature, 2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propylurea (200 mg, 0.4 mmol, 1.0 equiv), acetic acid (1 mL), acetic anhydride (1 mL) and hydrobromic acid (1 mL) were added to an 8 mL sample vial. The resulting reaction mixture was stirred at 120 °C overnight. The reaction mixture was basified to pH = 6 with saturated aqueous sodium bicarbonate. The reaction mixture was extracted with dichloromethane and methanol (3 x 50 mL). The combined organic phases were washed with saturated brine (3 x 50 mL) and dried over anhydrous sodium sulfate. After filtration of the resulting mixture, the filtrate was concentrated under reduced pressure to give 2-({9,10-dihydroxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propylurea (180 mg, yield 95.4%).

[0527] LCMS(ESI): [M+H] + = 464.2 3.

[0529]

[0530] At room temperature, to a solution of 2-({9,10-dihydroxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propionamide (150 mg, 0.3 mmol, 1.0 equiv) in N,N-dimethylformamide (9 mL) was added potassium carbonate (268 mg, 1.8 mmol, 6.0 equiv) and iodomethane-d3 (234 mg, 1.5 mmol, 5.0 equiv). The resulting reaction mixture was stirred at room temperature overnight. The crude product was purified by reverse-phase column chromatography to give 2-{[9,10-bis(2H3)methoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl](2,4,6-trimethylphenyl)amino}propionamide (25.92 mg, yield 15.52%), with the following conditions: (Column: Xbridge BEH Shield RP18 5 μm, 30*150 mm; Mobile phase A: water (10 mmol / L ammonium bicarbonate), Mobile phase B: acetonitrile; Flow rate: 60 mL / min; Gradient: 35% B to 45% B in 7 min; Wavelength: 254 nm / 220 nm; RT1 (min): 6.23.

[0531] LCMS(ESI): [M+H] + = 498.30

[0532] 1 1H NMR (400 MHz, Methanol-d 4 ) δ 7.11 (d, J = 10.6 Hz, 2H), 6.93 (s, 1H), 6.63 (s, 1H), 5.40 (s, 1H), 4.60 (s, 2H), 4.49 (s, 1H), 4.20–4.01 (m, 2H), 2.97 (t, J = 6.5 Hz, 2H), 2.37 (s, 3H), 2.22 (s, 3H), 2.09 (s, 3H), 1.04–0.96 (m, 3H).

[0533] Example 17, Compound 17

[0534]

[0535] To a solution of 2-[(1-aminopropan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (60 mg, 0.18 mmol, 1.0 equiv) in DCM (2 mL) was added triethylamine (41 mg, 0.40 mmol, 3.0 equiv) and dimethyl dicarbonate (36 mg, 0.27 mmol, 2.0 equiv). After stirring for two hours, the mixture was concentrated by rotary evaporation under reduced pressure. The crude product (80 mg) was purified by Prep-TLC on a normal phase, with EA / PE = 1 / 3, to give methyl N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]carbamate (23.01 mg, yield 32.94%).

[0536] LCMS(ESI):[M+H] + = 507.25

[0537] 1 H NMR(400MHz,Methanol-d 4 )δ7.09(m,2H),6.90(s,1H),6.61(s,1H),5.38(s,1H),4.48–4.42(m,1H),4.20–4.02(m,2H),3.86(s,3H),3.64(s,6H),3.60–3.53(m,1H),3.48–3.38(m,1H),2.94(t,J = 6.5Hz,2H),2.35(s,3H),2.19(s,3H),2.09(s,3H),1.06(d,J = 6.9Hz,3H).

[0538] Example 18, Compound 18

[0539]

[0540] A solution of benzoic acid (76 mg, 0.62 mmol, 3.5 equiv) in DMF (1 mL) was added to 2-[(1-aminopropan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (80 mg, 0.18 mmol, 1.0 equiv), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (103 mg, 0.53 mmol, 3.0 equiv), and 1-hydroxybenzotriazole (72 mg, 0.53 mmol, 3.0 equiv). The mixture was stirred for 2 h. The mixture was concentrated under reduced pressure, and the crude product (100 mg) was purified by Prep-TLC with EA / PE = 1 / 5 to give N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]benzamide (18.91 mg, yield 19.11%).

[0541] LC-MS (ESI): [M+H] + = 553.3

[0542] 1 1H NMR (400 MHz, Methanol-d 4 ) δ 7.92–7.87 (m, 2H), 7.59–7.45 (m, 3H), 7.17–7.08 (m, 2H), 6.91 (s, 1H), 6.63 (s, 1H), 5.42 (s, 1H), 4.69–4.60 (m, 1H), 4.19–4.02 (m, 2H), 3.93–3.82 (m, 4H), 3.79–3.72 (m, 1H), 3.64 (s, 3H), 2.96 (t, J = 6.5 Hz, 2H), 2.36 (s, 3H), 2.25 (s, 3H), 2.11 (s, 3H), 1.12 (d, J = 7.0 Hz, 3H).

[0543] Example 19, Compound 19

[0544]

[0545] A solution of 2-pyridinecarboxylic acid (41 mg, 0.34 mmol, 1.0 equiv) in DMF (1 mL) was added with 2-[(1-aminopropan-2-yl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (100 mg, 0.22 mmol, 1.0 equiv), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (64 mg, 0.34 mmol, 1.5 equiv), and 1-hydroxybenzotriazole (45 mg, 0.34 mmol, 1.5 equiv). The mixture was stirred for 2 h. The mixture was concentrated under reduced pressure, and the crude product (100 mg) was purified by Prep-HPLC to obtain N-[2-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,4,6-trimethylphenyl)amino)propyl]pyridine-2-carboxamide (17.35 mg, yield 13.93%).

[0546] LCMS(ESI):[M+H] + =554.25

[0547] 1 1H NMR(400MHz,Methanol-d 4 )δ8.69–8.60(m,1H),8.06(d,J=7.7Hz,1H),8.02–7.90(m,1H),7.58–7.50(m,1H),7.15–7.09(m,2H),6.91(s,1H),6.62(s,1H),5.40(s,1H),4.83–4.76(m,1H),4.18–4.04(m,2H),3.86(s,3H),3.85(d,J=6.4Hz,2H),3.64(s,3H),2.95(t,J=6.5Hz,2H),2.36(s,3H),2.25(s,3H),2.12(s,3H),1.09(d,J=6.9Hz,3H).

[0548] Example 20, Compound 20

[0549]

[0550] 9,10 - Dimethoxy - 2 - [(1 - amino - 2 - propyl)(2,4,6 - trimethylphenyl)amino] - 6H,7H - pyrimido[4,3 - a]isoquinolin - 4 - 1(70 mg, 0.2 mmol, 1.0 equivalent) and pyrimidine - 4 - carboxylic acid (58 mg, 0.5 mmol, 3.0 equivalents) were dissolved in the solvent DMF (2 mL). At room temperature, HOBT (63 mg, 0.5 mmol, 3.0 equivalents) and EDCI (90 mg, 0.5 mmol, 3.0 equivalents) were added to the above mixture, and the mixture was stirred at room temperature for 2 h. The crude product was purified by Prep - HPLC under the following conditions (column: XBridge BEH C18 5μm, 30*150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: from 40% B to 47% B in 7 minutes; wavelength: 254 nm / 220 nm; RT1(min): 6.45), to obtain N - [2 - ({9,10 - dimethoxy - 4 - oxo - 6H,7H - pyrimido[4,3 - a]isoquinolin - 2 - yl}(2,4,6 - trimethylphenyl)amino)propyl]pyrimidine - 4 - carboxamide (19.00 mg, yield 21.34%).

[0551] LCMS(ESI): [M + H] + = 555.35

[0552] 1 H NMR(400 MHz, Methanol - d 4 ) δ 9.32(s, 1H), 9.03(d, J = 4.8 Hz, 1H), 8.16–8.02(m, 1H), 7.18–7.10(m, 2H), 6.94(s, 1H), 6.64(s, 1H), 5.43(s, 1H), 4.60(s, 1H), 4.23–4.08(m, 2H), 3.89(d, J = 6.5 Hz, 5H), 3.67(s, 3H), 2.98(t, J = 6.5 Hz, 2H), 2.38(s, 3H), 2.27(s, 3H), 2.14(s, 3H), 1.23–0.98(m, 3H).

[0553] Example 21, Compound 21

[0554]

[0555] 9,10 - Dimethoxy - 2 - [(1 - amino - 2 - propyl)(2,4,6 - trimethylphenyl)amino] - 6H,7H - pyrimido[4,3 - a]isoquinolin - 4 - 1(50 mg, 0.1 mmol, 1.0 equivalent) was dissolved in solvent DCM (3 mL). At room temperature, isocyanatocyclopropane (14 mg, 0.2 mmol, 1.5 equivalents) and triethylamine (34 mg, 0.3 mmol, 3.0 equivalents) were added to the above mixture, and the mixture was stirred at room temperature for 2 h. The crude product was purified by Prep - HPLC under the following conditions (column Xbridge BEH Shield RP18, 5 μm, 19 * 250 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: from 36% B to 50% B in 7 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 6.05 / 6.5), to obtain 3 - cyclopropyl - 1 - [2 - ({9,10 - dimethoxy - 4 - oxo - 6H,7H - pyrimido[4,3 - a]isoquinolin - 2 - yl}(2,4,6 - trimethylphenyl)amino)propyl]urea (17.19 mg, yield 28.92%).

[0556] LCMS(ESI): [M + H] + = 532.30

[0557] 1 H NMR(400 MHz, Methanol - d 4 ) δ 7.15–7.05(m, 2H), 6.93(s, 1H), 6.63(s, 1H), 5.40(s, 1H), 4.57–4.49(m, 1H), 4.17–4.04(m, 2H), 3.88(s, 3H), 3.75–3.60(m, 4H), 3.55–3.45(m, 1H), 2.96(t, J = 6.5 Hz, 2H), 2.57–2.48(m, 1H), 2.37(s, 3H), 2.23(s, 3H), 2.11(s, 3H), 1.09(d, J = 6.9 Hz, 3H), 0.79–0.67(m, 2H), 0.57–0.46(m, 2H).

[0558] Example 22, Compound 22

[0559]

[0560] 9,10 - Dimethoxy - 2 - [(1 - amino - 2 - propyl)(2,4,6 - trimethylphenyl)amino] - 6H,7H - pyrimido[4,3 - a]isoquinolin - 4 - 1(70 mg, 0.2 mmol, 1.0 eq) and triethylamine (47 mg, 0.5 mmol, 3.0 eq) were dissolved in solvent DCM (3 mL) at room temperature. N - Methylcarbamoyl chloride (22 mg, 0.2 mmol, 1.5 eq) was added to the above mixture at 0 °C, and the mixture was stirred at room temperature for 2 h. The crude product was purified by Prep - HPLC under the following conditions (column: XBridge BEH C18 5 μm, 30 * 150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: from 36% B to 48% B in 7 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 6.37), and 1 - [2 - ({9,10 - dimethoxy - 4 - oxo - 6H,7H - pyrimido[4,3 - a]isoquinolin - 2 - yl}(2,4,6 - trimethylphenyl)amino)propyl] - 3 - methylurea (16.82 mg, yield 21.25%) was obtained.

[0561] LCMS(ESI): [M + H] + = 506.30

[0562] 1 H NMR(400 MHz, Methanol - d 4 ) δ 7.16–7.07 (m, 2H), 6.93 (s, 1H), 6.63 (s, 1H), 5.39 (s, 1H), 4.60 (s, 1H), 4.46 (t, J = 7.6 Hz, 1H), 4.21–4.00 (m, 2H), 3.95–3.85 (m, 4H), 3.66 (s, 3H), 3.05–2.80 (m, 5H), 2.37 (s, 3H), 2.21 (s, 3H), 2.08 (s, 3H), 1.02 (d, J = 6.9 Hz, 3H).

[0563] Example 23, Compound 23 - 1 and Compound 23 - 2

[0564]

[0565] Preparation steps: 1.

[0567]

[0568] 4-Bromo-2,6-diisopropylaniline (5 g, 20 mmol, 1.0 equiv), cesium carbonate (20.0 g, 60 mmol, 3 equiv), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (1.6 g, 2 mmol, 0.1 equiv), methylboronic acid (5.8 g, 98 mmol, 5 equiv) and 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (1.6 g, 4 mmol, 0.2 equiv) were dissolved in 1,4-dioxane (90 mL) and water (15 mL), protected by nitrogen, and stirred at 100 °C overnight. Cooled to room temperature, dissolved in 100 mL of water, extracted with ethyl acetate (3×100 mL), the organic layer was washed with 100 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by normal-phase column chromatography under the following conditions (mobile phase A: dichloromethane, mobile phase B: methanol; flow rate: 100 mL / min; gradient: 0% B to 10% B in 30 min; wavelength: 254 nm;) to obtain 2,6-diisopropyl-4-methylaniline (2.1 g, yield 56.2%).

[0569] LCMS(ESI): [M+H] + = 192.17 2.

[0571]

[0572] Under nitrogen protection, at 90 °C, a solution of 2,6-diisopropyl-4-methylaniline (1 g, 5.2 mmol, 1 equiv) and 2-chloro-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (1.68 g, 5.8 mmol, 1.1 equiv) in isopropanol (2 mL) was stirred and reacted overnight. The resulting reaction solution was concentrated under reduced pressure, filtered, the filter cake was collected and washed with ethyl acetate (2×10 mL) to obtain 2-[(2,6-diisopropyl-4-methylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (1.0 g, yield 42.7%).

[0573] LCMS(ESI): [M+H] + = 448.25 3.

[0575]

[0576] 2-[(2,6-Diisopropyl-4-methylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (700 mg, 1.6 mmol, 1 equiv), cesium carbonate (1.6 g, 4.8 mmol, 3 equiv) and tert-butyl N-(2-bromopropyl)carbamate (560 mg, 2.3 mmol, 1.5 equiv) were dissolved in 1,4-dioxane (10 mL), protected by nitrogen, and stirred at 100 °C overnight. Cooled to room temperature, dissolved in 20 mL of water, extracted with ethyl acetate (3 × 10 mL), the organic layer was washed with 20 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse-phase column chromatography under the following conditions (column: C18; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 50% B to 80% B in 20 min; wavelength: 254 nm;) to obtain tert-butyl N-{2-[(2,6-diisopropyl-4-methylphenyl)({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl})amino]propyl}carbamate (600 mg, yield 63.4%).

[0577] LCMS(ESI): [M+H] + = 605.35 4.

[0579]

[0580] tert-Butyl N-{2-[(2,6-diisopropyl-4-methylphenyl)({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl})amino]propyl}carbamate (500 mg, 0.91 mmol, 1 equiv) was dissolved in a 1,4-dioxane solution of 10 mL hydrochloric acid (4 M), and stirred at room temperature for 1 hour. Concentrated under reduced pressure, dissolved in 10 mL of water, extracted with ethyl acetate (3 × 10 mL), the organic layer was washed with 20 mL of saturated sodium chloride, dried over anhydrous Na 2 SO 4 dried, filtered, and concentrated under reduced pressure to obtain 2-((1-aminopropan-2-yl)(2,6-diisopropyl-4-methylphenyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (300 mg, yield 73.3%).

[0581] LCMS(ESI): [M+H] + = 505.30 5.

[0583]

[0584] 2-((1-Aminopropan-2-yl)(2,6-diisopropyl-4-methylphenyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (300 mg, 0.59 mmol, 1.0 equiv.), triethylamine (180 mg, 1.8 mmol, 3 equiv.) and trimethylsilyl isocyanate (103 mg, 1.2 mmol, 2 equiv.) were dissolved in 10 mL of DCM and stirred at room temperature for 2 h. Concentrated under reduced pressure, dissolved in 10 mL of water, extracted with ethyl acetate (3×10 mL), and the organic layer was washed with 20 mL of saturated sodium chloride, anhydrous Na 2 SO 4 dried, filtered, and concentrated under reduced pressure. The crude product was purified by reverse-phase column chromatography under the following conditions (column: XBridge BEH C18 5 μm, 30*150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 43% B to 55% B, 8 min; wavelength: 254 / 220 nm; RT1 (min): 5.77) to obtain 1-(2-((2,6-Diisopropyl-4-methylphenyl)(9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)amino)propyl)urea (150 mg, yield 45.0%). 6.

[0586]

[0587] The racemate (150 mg) was purified by Chiral-SFC under the following conditions (column: CHIRALPAK IH, 2*25 cm, 5 μm; mobile phase A: methanol:dichloromethane = 1:1 - HPLC, mobile phase B: methyl tert-butyl ether (0.1% ethylenediamine) - HPLC; flow rate: 20 mL / min; gradient: isocratic; column temperature (°C): 35; pressure (bar): 100; wavelength: 220 nm; RT1 (min): 10; RT2 (min): 5; sample solvent: methanol - HPLC; injection volume: 1 mL) to obtain the product.

[0588] Compound 23-1 (isomer 1, RT1 (min): 10, 16.5 mg, yield 6.4%).

[0589] LCMS (ESI): [M+H] + = 548.30

[0590] 1 H NMR (400 MHz, Methanol-d 4)δ 7.25–7.18 (m, 2H), 6.94 (s, 1H), 6.61 (s, 1H), 5.45 (s, 1H), 4.48 (s, 1H), 4.15–4.08 (m, 2H), 3.89 (s, 3H), 3.63 (s, 3H), 3.11–2.93 (m, 4H), 2.75 (s, 1H), 2.44 (s, 3H), 1.36–1.25 (m, 8H), 1.17 (d, J = 6.8 Hz, 3H), 1.02 (d, J = 6.6 Hz, 4H).

[0591] Compound 23-2 (isomer 2, RT2 (min): 5, 16.42 mg, yield 6.4%).

[0592] LCMS (ESI): [M+H] + = 548.30

[0593] 1 1H NMR (400 MHz, Methanol-d 4 )δ 7.26–7.18 (m, 2H), 6.94 (s, 1H), 6.61 (s, 1H), 5.45 (s, 1H), 4.48 (s, 1H), 4.16–4.08 (m, 2H), 3.89 (s, 3H), 3.63 (s, 3H), 3.11–2.93 (m, 3H), 2.75 (s, 1H), 2.44 (s, 3H), 1.35–1.28 (m, 7H), 1.17 (d, J = 6.8 Hz, 3H), 1.02 (d, J = 6.6 Hz, 3H).

[0594] Example 24, Compound 24-1 and Compound 24-2

[0595]

[0596] Preparation steps: 1.

[0598]

[0599] At room temperature, dissolve 1,3-difluoro-5-methyl-2-nitrobenzene (200 mg, 1.1 mmol, 1 equiv) in MeOH (5 mL) in air, and then add a methanol solution of sodium methoxide (3 ml, 60%). React overnight at room temperature. Evaporate under reduced pressure, add 50 mL of water, extract with ethyl acetate (2 x 50 mL), wash the combined organic layers with saturated brine (2 x 100 mL), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain 1,3-dimethoxy-5-methyl-2-nitrobenzene (200 mg, crude). 2.

[0601]

[0602] At room temperature, in air, 1,3-dimethoxy-5-methyl-2-nitrobenzene (190 mg, 0.9 mmol, 1 equiv) and H 2 O (1 mL) were added to a solution of EtOH (5 mL), and then NH 4 Cl (154 mg, 3 mmol, 3 equiv) and Fe (8.5 mg, 0.2 mmol, 3 equiv) were added. The reaction was carried out at 80 °C for two hours. The resulting mixture was filtered, and the filter cake was washed with ethyl acetate (2 x 30 mL). The reaction was quenched with water at room temperature. The resulting mixture was extracted with ethyl acetate (2 x 100 mL). The combined organic layers were washed with saturated sodium chloride (2 x 150 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give 2,6-dimethoxy-4-methylaniline (150 mg, crude).

[0603] LCMS (ESI): [M+H] + = 168.2 3.

[0605]

[0606] At room temperature, in air, 2-chloro-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (273 mg, 0.9 mmol, 1.2 equiv) was dissolved in a solution of isopropanol (3 mL), and 2,6-dimethoxy-4-methylaniline (152 mg, 0.9 mmol, 1.2 equiv) was added. The resulting mixture was concentrated under reduced pressure to give 2-[(2,6-dimethoxy-4-methylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (300 mg, crude).

[0607] LCMS (ESI): [M+H] + = 424.2 4.

[0609]

[0610] 2-[(2,6-Dimethoxy-4-methylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (300 mg, 0.7 mmol, 1 equiv), N-(2-bromopropyl)carbamate tert-butyl ester (168 mg, 0.7 mmol, 1 equiv) and Cs 2 CO 3(461 mg, 1.4 mmol, 2 equiv) was dissolved in 1,4-dioxane (5 mL) solution, and XantPhos (81 mg, 0.1 mmol, 0.1 equiv) and Pd 2 (dba) 3 (64 mg, 0.1 mmol, 0.1 equiv) were added. The resulting mixture was stirred overnight under nitrogen at 100 °C. The filtrate was concentrated under reduced pressure, and the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, acetonitrile in water (10 mmol / L NH 4 HCO 3 ), gradient from 10% to 50% in 10 minutes; detector, UV 254 nm. Finally, tert-butyl N-{2-[(2,6-dimethoxy-4-methylphenyl)({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl})amino]propyl}carbamate (270 mg, 59% yield) was obtained.

[0611] LCMS (ESI): [M+H] + = 581.2 5.

[0613]

[0614] Under nitrogen at room temperature, tert-butyl N-{2-[(2,6-dimethoxy-4-methylphenyl)({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl})amino]propyl}carbamate (300 mg, 0.6 mmol, 1.0 equiv) was dissolved in 1,4-dioxane (2 mL), and a 1,4-dioxane (2 mL) solution of hydrochloric acid was added. The resulting mixture was stirred for 1.5 hours under nitrogen at room temperature. The reaction was quenched with water at room temperature. The mixture was neutralized to pH = 9 with saturated sodium carbonate solution. The resulting mixture was extracted with ethyl acetate (2 x 50 mL). The combined organic layers were washed with saturated brine (2 x 100 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH 2 Cl 2 / MeOH = 20 / 1) to give 2-[(1-aminopropan-2-yl)(2,6-dimethoxy-4-methylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (150 mg, 49% yield).

[0615] LCMS (ESI): [M+H] + = 481.2 6.

[0617]

[0618] 2-[(1-Aminopropan-2-yl)(2,6-dimethoxy-4-methylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (400 mg, 0.8 mmol, 1 equiv) and TEA (0.4 mL, 2.5 mmol, 3 equiv) were added to DCM (4 mL), and isocyano trimethylsilane (191 mg, 1.6 mmol, 2.0 equiv) was added to the solution. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 h, and the reaction mixture was concentrated under reduced pressure. The crude product (420 mg) was purified by high performance liquid chromatography under the following conditions (column: Sunfire C18 5 μm, 30 mm × 4.5 mm): mobile phase A: water (0.1% FA), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: from 51% B to 74% B in 10 min; wavelength: 254 nm / 220 nm; RT1 (min): 6.03; 6.62), to give 1-(2-((2,6-dimethoxy-4-methylphenyl)(9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)amino)propyl)urea (130 mg), which was further purified by Chiral-HPLC under the following conditions (column: JW-CHIRALPAK IG-3, 3.0*50 mm; 3 μm; mobile phase A: MeOH:DCM = 1:1 - HPLC, mobile phase B: MtBE (0.1% DEA)-HPLC; flow rate: 20 mL / min; gradient: isocratic; wavelength: 220 nm; RT1 (min): 12; RT2 (min): 16.5; sample solvent: sample solvent: MeOH:DCM = 1:1 - HPLC; injection volume: 0.7 mL; number of runs: 5 times) to give the product.

[0619] Compound 24-1 (isomer 1, RT1 (min): 12, 30.12 mg, 6.8% yield)

[0620] LCMS (ESI): [M+H] + = 524.25

[0621] 1 H NMR (400 MHz, Methanol-d 4)δ 6.92 (s, 1H), 6.72–6.66 (m, 3H), 5.53 (s, 1H), 4.11–4.05 (m, 2H), 3.89 (s, 3H), 3.81 (d, J = 10.1 Hz, 6H), 3.69 (s, 3H), 2.95 (t, J = 6.5 Hz, 2H), 2.47 (s, 3H).

[0622] Compound 24-2 (isomer 2, RT2 (min) 16.5, 44.03 mg, 10% yield)

[0623] LCMS (ESI): [M+H] + = 524.25

[0624] 1 1H NMR (400 MHz, Methanol-d 4 )δ 6.92 (s, 1H), 6.72–6.66 (m, 3H), 5.53 (s, 1H), 4.11–4.05 (m, 2H), 3.89 (s, 3H), 3.81 (d, J = 10.1 Hz, 6H), 3.69 (s, 3H), 2.95 (t, J = 6.5 Hz, 2H), 2.47 (s, 3H).

[0625] Example 25, Compound 25-1 and Compound 25-2

[0626]

[0627] Preparation steps: 1.

[0629]

[0630] A mixture of 2,6-difluoro-4-methylaniline (1.0 g, 6.9 mmol, 1 equiv) and 2-chloro-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (2.6 g, 9.0 mmol, 1.3 equiv) in IPA (40 mL) was stirred overnight under a nitrogen atmosphere at 90 °C. The resulting mixture was concentrated under reduced pressure. It was basified to pH = 8 with saturated NaHCO 3 3. The resulting mixture was extracted with EtOEt (3 x 50 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous Na 2 2SO 4 4. The filtered filtrate was concentrated under reduced pressure. It was purified by silica gel column chromatography using CH 2 2Cl 2Elution with / MeOH(80 / 1 - 60 / 1) gave 2-[(2,6-difluoro-4-methylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (2.0 g, 71.6%).

[0631] LCMS(ESI): [M+H] + = 400.15 2.

[0633]

[0634] 2-[(2,6-Difluoro-4-methylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (440 mg, 1.1 mmol, 1 equiv) and tert-butyl N-(2-bromopropyl)carbamate (288.5 mg, 1.2 mmol, 1.1 equiv) were stirred in DMF (15 mL) and stirred overnight under a nitrogen atmosphere at 100 °C. The resulting mixture was concentrated in vacuo. It was purified by silica gel column chromatography and eluted with CH 2 Cl 2 / MeOH(10 / 1) to give tert-butyl N-{2-[(2,6-difluoro-4-methylphenyl)({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl})amino]propyl}carbamate (200 mg, 32.6%).

[0635] LCMS(ESI): [M+H] + = 557.25 3.

[0637]

[0638] N-{2-[(2,6-Difluoro-4-methylphenyl)({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-A]isoquinolin-2-yl})amino]propyl}carbamate (200 mg, 0.359 mmol, 1 equiv) was stirred in 1,4-dioxane (15 mL) under a nitrogen atmosphere at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. As a result, 2-[(1-aminoprop-2-en-2-yl)(2,6-difluoro-4-methylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (150 mg, 91.45%) was obtained. The crude product was used directly in the next step without further purification.

[0639] LCMS(ESI): [M+H] + = 457.2 4.

[0641]

[0642] 2-[(1-Aminoprop-2-en-2-yl)(2,6-difluoro-4-methylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (50 mg, 0.11 mmol, 1.0 eq) and trimethylsilyl isocyanate (12.6 mg, 0.11 mmol, 1.0 eq) were stirred in DCM (5 mL) at room temperature under a nitrogen atmosphere overnight. The resulting mixture was concentrated under reduced pressure. The crude product (60 mg) was purified by Prep-HPLC under the following conditions (column: XSelect CSH Fluoro Phenyl 5 μm, 19 mm * 250 mm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 42% B to 65% B, 10 min; wavelength: 254 nm / 220 nm; RT1 (min): 5.67) to give (2R)-2-[(2,6-Difluoro-4-methylphenyl)({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl})amino]propylurea (50 mg). It was further purified by Chiral-HPLC under the following conditions (column: CHIRALPAK IH 2 * 25 cm, 5 μm; mobile phase A: MeOH:DCM = 1:1 - HPLC, mobile phase B: MtBE (0.1% DEA) - HPLC; flow rate: -20 mL / min; gradient: isocratic; wavelength: 220 nm; RT1 (min): 11.2; RT2 (min): 15.0; sample solvent: MeOH:DCM = 1:1 - HPLC; injection volume: 1.5 mL; number of runs: 5) to give the product.

[0643] Compound 25-1 (isomer 1, RT1 (min): 11.2, 13.41 mg, 24.49%).

[0644] LCMS (ESI): [M + H] + = 500.25

[0645] 1 H NMR (400 MHz, Methanol-d 4)δ6.90(s,1H),6.87(s,1H),6.85(s,1H),6.83(s,1H),5.70(t,J=1.7Hz,1H),5.63(s,1H),4.00(t,J=6.3Hz,2H),3.96–3.90(m,1H),3.88(s,3H),3.73(s,3H),3.65–3.56(m,1H),2.95(t,J=6.2Hz,2H),2.35(s,3H),1.54(d,J=6.9Hz,3H).

[0646] Compound 25-2 (isomer 2, RT2 (min): 15.0, 15.09 mg, 21.59%)

[0647] LCMS(ESI): [M+H] + = 500.25

[0648] 1 H-NMR(400 MHz, Methanol-d 4 )δ6.89(s,1H),6.87(s,1H),6.86–6.84(m,1H),6.84–6.81(m,1H),5.70(t,J=1.6Hz,1H),5.63(s,1H),4.00(t,J=6.3Hz,2H),3.96–3.90(m,1H),3.88(s,3H),3.73(s,3H),2.97–2.93(m,2H),2.35(s,3H),1.54(d,J=6.9Hz,3H).

[0649] Example 26, Compound 26

[0650]

[0651] Preparation steps 1.

[0653]

[0654] In a single-necked flask, (2E)-9,10-dimethoxy-2-[(2,4,6-trimethylphenyl)imino]-3H,6H,7H-pyrimido[4,3-a]isoquinolin-4-one (1.0 g, 2.6 mmol, 1.0 equiv), dioxane (100 mL), cesium carbonate (1.7 g, 5.1 mmol, 2.0 equiv), and tert-butyl (N-{4-[(4-methylphenylsulfonyl)oxy]cyclohexyl}carbamate) (1.4 g, 3.8 mmol, 1.5 equiv) were added successively, and the reaction was carried out overnight at 100 °C. After filtration, the filtrate was concentrated, and the resulting residue was purified by reverse-phase column chromatography under the following conditions: the chromatography column was C18, the mobile phase was water and acetonitrile, with a 10% to 100% gradient for 30 minutes, and a UV detector at 220 nm. tert-Butyl (4-((9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)(methylsulfonyl)amino)cyclohexyl)carbamate (110 mg, 7.3%) was obtained.

[0655] LCMS(ESI,m / z):[M+H] + =589.3 2.

[0657]

[0658] In a single-necked flask, tert-butyl (4-((9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)(methylsulfonyl)amino)cyclohexyl)carbamate (100 mg, 0.2 mmol, 1.0 equiv) and a 1,4-dioxane solution of HCl (4 M, 15 mL) were added successively, and the reaction was carried out at room temperature for 2 h. After concentrating the solvent, 2-[(4-aminocyclohexyl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (80 mg, 96.4%) was obtained.

[0659] LCMS(ESI,m / z):[M+H] + =489.3 3.

[0661]

[0662] 2-[(4-Aminocyclohexyl)(2,4,6-trimethylphenyl)amino]-9,10-dimethoxy-6H,7H-pyrimido[4,3-a]isoquinolin-4-one (140 mg, 0.3 mmol, 1.0 equiv) was dissolved in solvent DCM (8 mL). Triethylamine (145 mg, 1.5 mmol, 5.0 equiv) and trimethylsilyl isocyanate (40 mg, 0.4 mmol, 1.2 equiv) were added to the above mixture at room temperature, and the mixture was stirred at room temperature for 3 h. The crude product was purified by Prep-HPLC under the following conditions (column: Xbridge BEH Shield RP18, 5 μm, 19 * 250 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 mL / min; gradient: from 39% B to 4% B in 10 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 9.4) to obtain 4-({9,10-dimethoxy-4-oxo-6H,7H-pyrimido[4,3-a]isoquinolin-2-yl}(2,6-dimethylphenyl)amino)cyclohexylurea (35.71 mg, yield 22.81%).

[0663] LCMS (ESI): [M + H]+ = 532.25

[0664] 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.10 (s, 2H), 6.92 (s, 1H), 6.62 (s, 1H), 5.39 (s, 1H), 4.18–4.03 (m, 3H), 3.88 (s, 4H), 3.66 (s, 3H), 2.96 (t, J = 6.4 Hz, 2H), 2.36 (s, 3H), 2.21 (s, 6H), 2.05–1.92 (m, 4H), 1.88–1.80 (m, 2H), 1.75–1.62 (m, 2H).

[0665] Example 27, Compound 27

[0666]

[0667] Preparation procedure 1.

[0669]

[0670] In a single-necked flask, (2E)-9,10-dimethoxy-2-[(2,4,6-trimethylphenyl)imino]-3H,6H,7H-pyrimido[4,3-a]isoquinolin-4-one (1.0 g, 2.6 mmol, 1.0 equiv), 1,4-dioxane (100 mL), cesium carbonate (2.5 g, 7.7 mmol, 3.0 equiv), and tert-butyl 4-[(4-methylbenzenesulfonyl)oxy]piperidine-1-carboxylate (1.0 g, 2.8 mmol, 1.1 equiv) were added successively, and the reaction was carried out overnight at 100 °C. The mixture was filtered, and the filtrate was concentrated. The resulting residue was purified by reverse-phase column chromatography under the following conditions: mobile phase, water and acetonitrile, 10% to 100% gradient for 30 minutes, UV detector at 220 nm, to obtain tert-butyl 4-((9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)(formamidinyl)amino)piperidine-1-carboxylate (120 mg, yield 8.17%).

[0671] LCMS(ESI,m / z):[M+H] + =575.3 2.

[0673]

[0674] In a single-necked flask, tert-butyl 4-((9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)(formamidinyl)amino)piperidine-1-carboxylate (100 mg, 0.2 mmol, 1.0 equiv) and 1,4-dioxane solution of hydrogen chloride (17 mL) were added successively, and the reaction was carried out at room temperature for 2 h. The solvent was concentrated to obtain 2-(methylsulfonyl(piperidin-4-yl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (80 mg, yield 96.9%).

[0675] LCMS(ESI,m / z):[M+H] + =475.3 3.

[0677]

[0678] To a solution of 2-(Methylsulfonyl(piperidin-4-yl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (1050 mg, 2.2 mmol, 1.0 equiv) in dichloromethane (20 mL) was added triethylamine (640 mg, 6.3 mmol, 3.0 equiv) and trimethylsilyl isocyanate (364 mg, 3.2 mmol, 1.5 equiv). The mixture was stirred at room temperature for 2 h. After the mixture was concentrated by rotary evaporation under reduced pressure, the resulting residue was purified by reverse-phase column chromatography under the following conditions: Column: XBridge BEH Shield RP18, 5 μm, 19*250 mm; Mobile phase A: water (10 mmol / L ammonium bicarbonate), Mobile phase B: acetonitrile; Flow rate: 25 mL / min; Gradient: 35% B to 40% B in 10 min; Wavelength: 254 nm / 220 nm; RT1 (min): 8.57, to give 4-[(2E)-9,10-dimethoxy-4-oxo-2-[(2,4,6-trimethylphenyl)imino]-6H,7H-pyrimido[4,3-a]isoquinolin-3-yl]piperidine-1-carboxamide) (39.74 mg, yield 3.63%).

[0679] LCMS(ESI): [M+H] + = 518.40

[0680] 1 H NMR(400 MHz, Methanol-d 4 ) δ 7.10 (s, 2H), 6.92 (s, 1H), 6.62 (s, 1H), 5.39 (s, 1H), 4.73–4.62 (m, 1H), 4.18–3.97 (m, 4H), 3.88 (s, 3H), 3.66 (s, 3H), 3.02–2.90 (m, 4H), 2.36 (s, 3H), 2.17 (s, 8H), 1.43–1.29 (m, 2H).

[0681] Example 28, Compound 28

[0682]

[0683] The crude product (150 mg) was purified by Chiral-SFC under the following conditions (chromatographic column: CHIRALPAK IH, 2 * 25 cm, 5 μm; mobile phase A: methanol:dichloromethane = 1:1 - HPLC, mobile phase B: methyl tert-butyl ether (0.1% ethylenediamine) - HPLC; flow rate: 20 mL / min; gradient: isocratic; wavelength: 220 nm; RT1 (min): 8; RT2 (min): 13.0; sample solvent: methanol - HPLC; injection volume: 1 mL) to obtain 1-(3-((2,6-diisopropyl-4-methylphenyl)(9,10-dimethoxy-4-oxo-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-2-yl)amino)cyclobutyl)urea (13.00 mg, yield 14.9%).

[0684] LCMS(ESI): [M+H] + = 560.30

[0685] 1 H NMR(400MHz, Methanol-d 4 ) δ 7.21(s, 2H), 6.93(s, 1H), 6.64(s, 1H), 5.46(s, 1H), 4.51–4.44(m, 1H), 4.14–4.06(m, 2H), 3.97–3.88(m, 1H), 3.89(s, 3H), 3.64(s, 3H), 2.94 - 2.90(m, 4H), 2.74 - 2.78(m, 2H), 2.44(s, 3H), 2.20–2.08(m, 2H), 1.30(d, J = 6.8Hz, 7H), 1.11(d, J = 6.8Hz, 6H).

[0686] Example 29, Compound 29

[0687]

[0688] The crude product (30 mg) was purified by Chiral-HPLC under the following conditions: CHIRALPAK IH 3 * 25 cm, 5 μm; mobile phase A: CO 2 , mobile phase B: methanol (0.1% diethylamine); flow rate: 85 mL / min; gradient: isocratic 35% B; column temperature (°C): 25°C; back pressure (bar): 100; wavelength: 254 nm; RT1 (min): 2.85; RT2 (min): 4.5 Sample solvent: methanol:dichloromethane = 1:1 - HPLC; injection volume: 1 mL) to obtain the product.

[0689] Compound 29 (isomer 1, RT1 (min): 2.85, 3.2 mg, yield 10.3%)

[0690] LCMS (ESI, m / z): [M+H] + = 558.35

[0691] 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.17–7.10 (m, 2H), 6.93 (s, 1H), 6.65 (s, 1H), 5.43 (s, 1H), 4.78–4.69 (m, 1H), 4.20–4.04 (m, 2H), 3.97–3.77 (m, 5H), 3.67 (s, 3H), 2.98 (t, J = 6.5 Hz, 2H), 2.59–2.52 (m, 3H), 2.38 (s, 3H), 2.27 (s, 3H), 2.13 (s, 3H), 1.11 (d, J = 6.9 Hz, 3H).

[0692] Example 30, Compound 30

[0693]

[0694] At room temperature, 2-((1-aminopropan-2-yl)(trimethyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (100 mg, 0.22 mmol), 6-methylpyridine-2-carboxylic acid (45.3 mg, 0.33 mmol), EDCI (63.3 mg, 0.33 mmol), HOBT (44.6 mg, 0.33 mmol) and DMF (2 mL) were added to a 25 mL single-necked flask and stirred at room temperature for 24 h. After the reaction was completed, the reaction system was filtered and purified by reverse-phase Flash (mobile phase A: 0.5% FA aqueous solution, mobile phase B: acetonitrile; flow rate: 40 mL / min; gradient: from 0% B to 55% B within 20 minutes; the product eluted at 50% B, wavelength: 254 nm / 220 nm;) to obtain the product (19.2 mg, 10.3%).

[0695] LC-MS (ESI): [M+H] + = 568.2

[0696] 1 H NMR (400 MHz, CDCl 3)8.76 - 8.74(m, 1H), 7.86(d, J = 8.0Hz, 1H), 7.63(t, J = 12.0Hz, 1H), 7.19 - 7.16(m, 1H), 6.91(d, J = 8.0Hz, 1H), 6.62(s, 1H), 6.52(s, 1H), 5.22(s, 1H), 4.96 - 4.95(m, 1H), 4.16–4.10(m, 2H), 3.83(s, 3H), 3.82 - 3.75(m, 2H), 3.63(s, 3H), 2.84(t, J = 8.0Hz, 2H), 2.54(s, 3H), 2.26(s, 3H), 2.17(s, 3H), 2.05(s, 3H), 1.05(d, J = 8.0Hz, 3H).

[0697] Example 31, Compound 31

[0698]

[0699] At room temperature, 2 - ((1 - aminopropan - 2 - yl)(trimethyl)amino)-9,10 - dimethoxy - 6,7 - dihydro - 4H - pyrimido[6,1 - a]isoquinolin - 4 - one (150 mg, 0.33 mmol), 5 - methylpyridine - 2 - carboxylic acid (68.6 mg, 0.50 mmol), EDCI (95.9 mg, 0.50 mmol), HOBT (67.6 mg, 0.50 mmol) and DMF (3 mL) were added to a 25 mL single - necked flask and stirred at room temperature for 24 h. After the reaction was completed, the reaction system was filtered and purified by reverse - phase Flash (mobile phase A: 0.5% FA aqueous solution, mobile phase B: acetonitrile; flow rate: 40 mL / min; gradient: from 0% B to 55% B in 20 minutes; the product eluted at 50% B, wavelength: 254 nm / 220 nm;) to obtain the product (37.5 mg, 19.7%).

[0700] LC - MS(ESI): [M + H] + = 568.2

[0701] 1 H NMR(400 MHz, CDCl 3)8.81(s,1H),8.35(s,1H),7.95(d,J=8.0Hz,1H),7.54-7.51(m,1H),6.93(d,J=8.0Hz,1H),6.62(s,1H),6.52(s,1H),5.21(s,1H),4.92-4.87(m,1H),4.17–4.07(m,1H),3.83(s,3H),3.82-3.75(m,2H),3.63(s,3H),2.84(t,J=8.0Hz,2H),2.31(s,3H),2.25(s,3H),2.17(s,3H),2.05(s,3H),1.09(d,J=8.0Hz,3H).

[0702] Example 32, Compound 32

[0703]

[0704] At room temperature, 2-((1-aminopropan-2-yl)(methyltrimethyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (150 mg, 0.33 mmol), 4-methyl-2-pyridinecarboxylic acid (68.6 mg, 0.50 mmol), EDCI (95.9 mg, 0.50 mmol), HOBT (67.6 mg, 0.50 mmol) and DMF (3 mL) were added to a 25 mL single-necked flask and stirred at room temperature for 24 h. After the reaction was completed, the reaction system was filtered and purified by reverse-phase Flash (mobile phase A: 0.5% aqueous FA solution, mobile phase B: acetonitrile; flow rate: 40 mL / min; gradient: from 0% B to 55% B in 20 minutes; product eluted at 50% B, wavelength: 254 nm / 220 nm;) to obtain the product (32.3 mg, 17.3%).

[0705] LC-MS(ESI):[M+H] + =568.2

[0706] 1 H NMR(400MHz,CDCl 3)8.87(s,1H),8.37(d,J=8.0Hz,1H),8.21(s,1H),7.91(s,1H),7.15(d,J=8.0Hz,1H),6.91(d,J=12Hz,1H),6.62(s,1H),6.51(s,1H),5.23(s,1H),4.87-4.82(m,1H),4.19–4.07(m,2H),3.83(s,3H),3.82-3.75(m,2H),3.64(s,3H),2.85(t,J=8.0Hz,2H),2.34(s,3H),2.26(s,3H),2.17(s,3H),1.98(s,3H),1.07(d,J=4.0Hz,3H).

[0707] Example 33, Compound 33

[0708]

[0709] At room temperature, 2-((1-aminopropan-2-yl)(methyltrimethyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (150 mg, 0.33 mmol), 3-methyl-2-pyridinecarboxylic acid (68.6 mg, 0.50 mmol), EDCI (95.9 mg, 0.50 mmol), HOBT (67.6 mg, 0.50 mmol) and DMF (3 mL) were added to a 25 mL single-necked flask and stirred at room temperature for 24 h. After the reaction was completed, the reaction system was filtered and purified by reverse-phase Flash (mobile phase A: 0.5% aqueous FA solution, mobile phase B: acetonitrile; flow rate: 40 mL / min; gradient: from 0% B to 55% B in 20 minutes; the product eluted at 50% B, wavelength: 254 nm / 220 nm;) to obtain the product (12.8 mg, 6.8%).

[0710] LC-MS(ESI):[M+H] + =568.2

[0711] 1 H NMR(400MHz,CDCl 3)8.86(s,1H),8.35(d,J=4.0Hz,1H),7.47(d,J=8.0Hz,1H),7.22-7.20(m,1H),6.91(d,J=8.0Hz,1H),6.62(s,1H),6.52(s,1H),5.22(s,1H),4.96-4.91(m,1H),4.19–4.03(m,2H),3.83(s,3H),3.82-3.75(m,2H),3.63(s,3H),2.84(t,J=8.0Hz,2H),2.64(s,3H),2.26(s,3H),2.18(s,3H),2.02(s,3H),1.08(d,J=4.0Hz,3H).

[0712] Example 34, Compound 34

[0713]

[0714] At room temperature, 2-((1-aminopropan-2-yl)(methyltrimethyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (100 mg, 0.22 mmol), pyrimidine-2-carboxylic acid (40.9 mg, 0.33 mmol), EDCI (63.3 mg, 0.33 mmol), HOBT (44.6 mg, 0.33 mmol) and DMF (2 mL) were added to a 25 mL single-necked flask and stirred at room temperature for 24 h. After the reaction was completed, the reaction system was filtered and purified by reverse-phase Flash (mobile phase A: 0.5% aqueous FA solution, mobile phase B: acetonitrile; flow rate: 40 mL / min; gradient: from 0% B to 55% B in 20 minutes; the product eluted at 50% B, wavelength: 254 nm / 220 nm;) to obtain the product (20 mg, 10.9%).

[0715] LC-MS(ESI):[M+H] + =555.2

[0716] 11H NMR (400 MHz, DMSO-d6) δ 9.56 (t, J = 8.0 Hz, 1H), 9.03 - 9.00 (m, 2H), 8.16 (s, 1H), 7.87 (s, 1H), 7.74 - 7.71 (m, 1H), 7.12 - 7.10 (m, 2H), 7.02 (s, 1H), 4.74–4.70 (m, 1H), 4.16 (s, 3H), 4.15 - 4.10 (m, 2H), 3.98 - 3.95 (m, 2H), 3.89 (s, 3H), 3.05 - 3.00 (m, 2H), 2.31 (s, 3H), 2.19 (s, 3H), 2.03 (s, 3H), 0.83 (d, J = 8.0 Hz, 3H).

[0717] Example 35, Compound 35

[0718]

[0719] At room temperature, 2-((1-aminopropan-2-yl)(trimethyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (100 mg, 0.22 mmol), 5-methylpyrimidine-2-carboxylic acid (45.6 mg, 0.33 mmol), EDCI (63.3 mg, 0.33 mmol), HOBT (44.6 mg, 0.33 mmol) and DMF (2 mL) were added to a 25 mL single-necked flask and stirred at room temperature for 24 h. After the reaction was completed, the reaction system was filtered and purified by reverse-phase Flash (mobile phase A: 0.5% aqueous FA solution, mobile phase B: acetonitrile; flow rate: 40 mL / min; gradient: from 0% B to 55% B in 20 minutes; product eluted at 50% B, wavelength: 254 nm / 220 nm;) to obtain the product (27.6 mg, 14.7%).

[0720] LC-MS (ESI): [M+H] + = 569.2

[0721] 1 1H NMR (400 MHz, CDCl 3)9.28(s,1H),8.64(s,1H),6.91(d,J = 8.0Hz,1H),6.62(s,1H),6.51(s,1H),5.22(s,1H),4.93 - 4.90(m,1H),4.13–4.10(m,2H),3.83(s,3H),3.82 - 3.75(m,2H),3.63(s,3H),2.85 - 2.83(m,2H),2.31(s,3H),2.26(s,3H),2.16(s,3H),2.04(s,3H),1.05(d,J = 8.0Hz,3H).

[0722] Example 36, Compound 36

[0723]

[0724] At room temperature, 2-((1-aminopropan-2-yl)(trimethyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (100 mg, 0.22 mmol), pyrazine-2-carboxylic acid (40.9 mg, 0.33 mmol), EDCI (63.3 mg, 0.33 mmol), HOBT (44.6 mg, 0.33 mmol) and DMF (2 mL) were added to a 25 mL single-necked flask and stirred at room temperature for 24 h. After the reaction was completed, the reaction system was filtered and purified by reverse-phase Flash (mobile phase A: 0.5% FA aqueous solution, mobile phase B: acetonitrile; flow rate: 40 mL / min; gradient: from 0% B to 55% B in 20 minutes; product eluted at 50% B, wavelength: 254 nm / 220 nm;) to obtain the product (9.5 mg, 5.2%).

[0725] LC-MS(ESI):[M + H] + = 555.2

[0726] 11H NMR (400 MHz, DMSO-d6) δ 9.51 (t, J = 8.0 Hz, 1H), 9.13 - 9.12 (m, 1H), 8.92 (d, J = 4.0 Hz, 1H), 8.78 - 8.77 (m, 1H), 7.98 (s, 1H), 7.60 (s, 1H), 7.10 - 7.08 (m, 1H), 7.05 (s, 1H), 7.02 (s, 1H), 4.74–4.70 (m, 1H), 4.08 (s, 3H), 4.07 - 4.05 (m, 2H), 3.98 - 3.95 (m, 2H), 3.89 (s, 3H), 3.05 - 3.00 (m, 2H), 2.31 (s, 3H), 2.19 (s, 3H), 2.03 (s, 3H), 0.83 (d, J = 8.0 Hz, 3H).

[0727] Example 37, Compound 37

[0728]

[0729] At room temperature, 2-((1-aminopropan-2-yl)(trimethyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (100 mg, 0.22 mmol), 6-methylpyrazine-2-carboxylic acid (45.6 mg, 0.33 mmol), EDCI (63.3 mg, 0.33 mmol), HOBT (44.6 mg, 0.33 mmol) and DMF (2 mL) were added to a 25 mL single-necked flask and stirred at room temperature for 24 h. After the reaction was completed, the reaction system was filtered and purified by reverse-phase Flash (mobile phase A: 0.5% aqueous FA solution, mobile phase B: acetonitrile; flow rate: 40 mL / min; gradient: from 0% B to 55% B in 20 minutes; product eluted at 50% B, wavelength: 254 nm / 220 nm;) to obtain the product (29.3 mg, 15.6%).

[0730] LC-MS (ESI): [M+H] + = 569.2

[0731] 1 1H NMR (400 MHz, CDCl 3)9.17(s,1H),8.97(s,1H),8.59(s,1H),7.00(d,J=8.0Hz,2H),6.70(s,1H),6.60(s,1H),5.31(s,1H),5.08 - 5.07(m,1H),4.23–4.17(m,2H),3.92(s,3H),3.92 - 3.85(m,2H),3.72(s,3H),2.94 - 2.91(m,2H),2.68(s,3H),2.34(s,3H),2.25(s,3H),2.12(s,3H),1.11(d,J=8.0Hz,3H).

[0732] Example 38, Compound 38

[0733]

[0734] At room temperature, 2-((1-aminopropan-2-yl)(trimethyl)amino)-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,1-a]isoquinolin-4-one (150 mg, 0.33 mmol), 5-methylpyrazine-2-carboxylic acid (69.1 mg, 0.50 mmol), EDCI (95.8 mg, 0.50 mmol), HOBT (67.6 mg, 0.50 mmol) and DMF (3 mL) were added to a 25 mL single-necked flask and stirred at room temperature for 24 h. After the reaction was completed, the reaction system was filtered and purified by reverse-phase Flash (mobile phase A: 0.5% aqueous FA solution, mobile phase B: acetonitrile; flow rate: 40 mL / min; gradient: from 0% B to 55% B in 20 minutes; the product eluted at 50% B, wavelength: 254 nm / 220 nm;) to obtain the product (50.2 mg, 26.8%).

[0735] LC-MS(ESI):[M + H] + = 569.2

[0736] 1 H NMR(400MHz,CDCl 3)9.23(s,1H),8.48(s,1H),8.59(s,1H),7.00(d,J=12.0Hz,2H),6.70(s,1H),6.60(s,1H),5.31(s,1H),4.98 - 4.93(m,1H),4.23–4.17(m,2H),3.92(s,3H),3.92 - 3.85(m,2H),3.72(s,3H),2.94 - 2.91(m,2H),2.65(s,3H),2.35(s,3H),2.25(s,3H),2.12(s,3H),1.14(d,J=8.0Hz,3H).

[0737] PDE Enzyme Activity Inhibition Experiment

[0738] The compounds of each example were tested for PDE enzyme activity inhibition experiment by the FP method, and the compound RPL - 554 in Example 1 of CN100415743C was used as the positive control.

[0739]

[0740] Prepare PDE enzyme and Substrate (FAM - cyclic adenosine monophosphate / cyclic adenosine monophosphate) solutions in the reaction buffer (1×IMAP Reaction Buffer containing 0.1% BSA supplemented with 1 mM DTT). The starting concentration of the positive control for PDE was 1 / 10 μM, diluted 3 - fold, 10 + 0 dose. 0.05 μL of the compound in 100% DMSO was delivered to a 384 - well plate (Corning 4514) by acoustic liquid delivery technology (Echo 655) and centrifuged at 1000 rpm for 1 minute; 2.5 μL of the PDE enzyme solution was transferred to the 384 - well reaction plate and centrifuged at 1000 rpm for 1 minute, and incubated at 25 °C for 10 min; 2.5 μL of the Sub solution was transferred to the 384 - well reaction plate, centrifuged at 1000 rpm for 1 minute, and incubated at 25 °C for 60 min; 15 μL of the binder mixture was transferred to the 384 - well reaction plate and centrifuged at 1000 rpm for 1 minute, and incubated at 25 °C for 60 min; finally, the FP signal was read using BMG (PHERAstar FSX). The IC50 value and non - linear regression curve fitting were obtained using GraphPad Prism software.

[0741] Table 3 shows the IC 50 values of the enzyme activity experiments of the compounds in the comparative examples and examples. The smaller the IC 50 value, the less amount of the compound required for half - inhibition of different PDE enzymes, indicating stronger inhibitory activity.

[0742] Table 3

[0743]

[0744]

[0745] Conclusion: The compounds of the present invention have high inhibitory activity against PDE3 / 4, which is superior to the prior art and has good development prospects.

[0746] The above are only specific embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A compound having the structural formula I or a pharmaceutically acceptable form thereof, wherein the pharmaceutically acceptable form is selected from pharmaceutically acceptable salts or co-crystals, stereoisomers, tautomers, deuterated compounds, solvates, chelates, non-covalent complexes or prodrugs: Wherein: R 1 、R 2 each independently selected from H, C 1-6 linear alkyl, C 3-6 branched alkyl and C 3-6 cycloalkyl; the linear alkyl, branched alkyl or cycloalkyl is optionally further substituted by 0 to 4 substituents selected from D, F, Cl, Br, I, OH, =O, NH 2 、CN, COOH, C 1-4 alkyl, C 1-4 alkoxy; R 3 、R 4 、R 5 are each independently selected from H, halogen, CN, C 1-6 alkoxy, C 1-6 linear alkyl, C 3-6 branched alkyl, and C 3-6 cycloalkyl; the alkoxy, linear alkyl, branched alkyl or cycloalkyl is optionally further substituted by 0 to 4 substituents selected from D, F, Cl, Br, I, OH, ═O, NH 2 , CN, COOH, C 1-4 alkyl, C 1-4 alkoxy; L is selected from and, n is 0, 1 or 2; k is 0, 1, 2 or 3; the H in the said L is optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, OH, =O, NH 2 , CN, COOH, C 1-4 alkyl, C 1-4 alkoxy; Among them, R 9 and R 10 are each independently selected from H, C 1-6 linear alkyl, C 3-6 branched alkyl, C 3-6 cycloalkyl, C 3-6 heterocycloalkyl, C 6-10 aryl, C 5-10 heteroaryl and COOCH 3 , and R 9 and R 10 are not simultaneously H; the heteroaryl contains 1-3 heteroatoms selected from N, O and S; R 6 selected from wherein, R 11 is selected from amino, C 1-6 alkoxy, C 3-12 cycloalkyl, C 6-10 aryl, C 5-10 heterocyclic group, the heterocyclic group contains 1 - 3 heteroatoms selected from N, O and S, and the amino, alkoxy, cycloalkyl, aryl, heteroaryl are optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, OH, =O, NH 2 , CN, COOH, CF 3 , C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl; Optionally, R 6 forms with L R 7 and R 8 are each independently selected from H, =O, halogen, NH 2 , CN, C 1-6 linear alkyl, C 3-6 branched alkyl, C 3-6 cycloalkyl and 2. The compound or a pharmaceutically acceptable form thereof according to claim 1, Wherein: R 1 、R 2 each independently selected from CH 3 、CHF 2 、CD 3 or C 3-6 cycloalkyl; Preferably, R 1 , R 2 are respectively CD 3 ; Preferably, R 1 , R 2 One of them is CH 3 , and the other is CHF 2 .

3. The compound or a pharmaceutically acceptable form thereof according to claim 1 or 2, Wherein: R 3 、R 4 、R 5 are each independently selected from CH 3 、i-Pr、OMe、CD 3 or halogen; Preferably, R 5 is CH 3 , and the CH 3 is optionally further substituted by 0 to 3 Ds; R 3 , R 4 are the same and are selected from CH 3 , i-Pr, OMe or halogen.

4. The compound or a pharmaceutically acceptable form thereof according to any one of claims 1-3, Wherein, L is selected from one of the following schemes: L is n is 0 or 1, preferably 0; Preferably, k is 0, 1 or 2; Preferably, R 9 , R 10 One of them is H, and the other is selected from H, C 1-6 linear alkyl, C 3-6 branched alkyl, C 3-6 cycloalkyl, C 3-6 heterocycloalkyl, C 6-10 aryl, C 5-10 heteroaryl and COOCH 3 ; the heteroaryl contains 1 or 2 heteroatoms N; the H in the L is optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, OH, =O, NH 2 , CN, COOH, C 1-4 alkyl, C 1-4 alkoxy; L is L is n is 0 or 1, and k is 1; L is n is 0 or 1, and k is 1; L is n is 2 and k is 2; L is n is 1, k is 1 or 2; or L is n is 2 and k is 2.

5. The compound or a pharmaceutically acceptable form thereof according to any one of claims 1-4, Wherein, R 6 selected from one of the following options: R 6 is wherein R 11 is an amino group, and the amino group is optionally further substituted by 0, 1 or 2 substituents selected from F, Cl, Br, I, OH, ═O, NH 2 , CN, COOH, CF 3 , C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl; R 6 is wherein R 11 is C 6-10 aryl or C 5-10 heterocyclic group, the heterocyclic group containing 1, 2 or 3 heteroatoms N; the aryl and heteroaryl are optionally further substituted by 0, 1, 2 or 3 substituents selected from H, F, Cl, Br, I, OH, =O, NH 2 , CN, COOH, CF 3 , C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl.

6. The compound or a pharmaceutically acceptable form thereof according to any one of claims 1-5, Wherein, R 7 is H or CH 3 ,R 8 is H or F.

7. The compound or a pharmaceutically acceptable form thereof according to claim 1, Wherein, The compound is selected from one or more of the compounds shown in Table 1.

8. An intermediate compound having the structure shown in formula II: Wherein, R 1 、R 2 、R 3 、R 4 、R 5 、R 7 、R 8 as defined in any one of claims 1 - 7; L 1 Selected from R 9 、R 10 、n, k are as defined in any one of claims 1 - 7; R 12 、R 13 are each independently H, Boc, Cbz, SEM, Fmoc, Alloc, Pht, OTs, PMB, Bn, Trt; Preferably, the intermediate compound is R 12 、R 13 One of them is H, and the other is Boc, Cbz, SEM, Fmoc, Alloc, Pht, OTs, PMB, Bn, Trt.

9. A method for preparing the compound or a pharmaceutically acceptable form thereof according to any one of claims 1-7, the method Comprises: Modify the R shown in the intermediate compound of formula II as claimed in claim 8 12 and R 13 at the terminal to prepare a compound having the structural formula I.

10. The method according to claim 9, the method further comprising a process for preparing the intermediate compound according to claim 8.

11. A pharmaceutical composition, which Comprises: The compound or a pharmaceutically acceptable form thereof according to any one of claims 1-7, and a pharmaceutically acceptable carrier, excipient and / or one or more other therapeutic agents.

12. Use of the compound or a pharmaceutically acceptable form thereof according to any one of claims 1-7 or the pharmaceutical composition according to claim 11 in the preparation of a preparation for inhibiting phosphodiesterase.

13. Use of the compound or a pharmaceutically acceptable form thereof according to any one of claims 1-7 or the pharmaceutical composition according to claim 11 in the preparation of a drug for treating phosphodiesterase-related diseases; Preferably, wherein the phosphodiesterase-related diseases include respiratory diseases such as asthma.

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