CN108341819B - Phosphodiesterase inhibitors and uses thereof - Google Patents

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a phosphodiesterase 9(PDE9) inhibitor shown as a formula I, pharmaceutically acceptable salts, solvate compounds, polymorphs and isomers thereof, and also relates to pharmaceutical preparations, pharmaceutical compositions and applications of the compounds. The compound and the pharmaceutically acceptable salt, the solvate, the polymorph and the isomer thereof can be applied to the treatment of related diseases mediated by the abnormal expression of phosphodiesterase 9(PDE 9).

Description

Phosphodiesterase inhibitors and uses thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a phosphodiesterase inhibitor shown in a formula I, and pharmaceutically acceptable salts, solvate compounds, polymorphs and isomers thereof, wherein the compound can be applied to treatment of related diseases mediated by abnormal expression of phosphodiesterase 9(PDE 9).

Background

Phosphodiesterases (PDEs) are a class of proteases that selectively degrade cGMP (cyclic guanosine monophosphate) and cAMP (cyclic adenosine monophosphate), important second messengers in the body, and thus participate in important physiological processes in the body. PDEs can be divided into 11 members (PDE1-PDE11) according to the sequence homology of genes and selectivity for cGMP or cAMP. Among them, PDE9A is an important member of the PDE family, which is widely expressed in testis, brain, small intestine, skeletal muscle, heart, lung, thymus and pancreas. With the intensive research in recent years, there have been numerous literature reports and clinical data demonstrating that PDE9A inhibitors are useful for the treatment of diseases in which cognitive impairment is caused by central nervous system disorders, such as senile dementia and schizophrenia, and neurodegenerative process diseases of the brain.

Two nucleotides, cAMP and cGMP, are important second messengers that play a central role in cell signaling; they mainly activate protein kinases: activated by cAMP is called protein kinase a (pka), and activated by cGMP is called protein kinase g (pkg). Activated PKA and PKG can phosphorylate a number of cellular effector proteins such as ion channels, G-protein coupled receptors, structural proteins, transduction factors. Thus, cAMP and cGMP in this way may control most physiological processes in many organs. Meanwhile, cAMP and cGMP can also act directly on effector proteins, thereby playing the same role as described above. It is well known that cGMP can act directly on ion receptors, thereby affecting the ion concentration in cells. Phosphodiesterases (PDEs) hydrolyze cyclic monophosphates cAMP and cGMP, converting them to the inactive monophosphates AMP and GMP.

Human PDE9A was first cloned and sequenced in 1998 and is the most selective PDE for cGMP reported to date. PDE9A has a binding constant (Km) for cGMP of 170nM, whereas the binding constant for cAMP is as high as 230000nM with a selectivity of over 1000-fold. In comparison to PDE2A and PDE5A, PDE9A inhibitors may increase baseline cGMP concentrations because PDE9A has no cGMP binding region and thus the catalytic activity of PDE9A is not enhanced by cGMP.

Conventional PDE inhibitors do not inhibit human PDE9A and therefore the drugs IBMX, dipyridamole, SKF94120, rolipram and vinpocetin have no inhibitory activity or very low activity against PDE 9A. There are no PDE9A inhibitor drugs on the market today, only a few inhibitors that are in the clinical development stage. Two classes of PDE9A inhibitors, PF-04447943 from Pfizer and BI-409306 from BI, respectively, have the following structural formulae:

two compounds are currently in phase one and phase two clinical stages. Both classes of PDE9A inhibitors are based on the parent 4-hydroxypyrazolo [3,4-d ] pyrimidine, although developed by two different companies.

Disclosure of Invention

The invention aims to provide a PDE9 kinase inhibitor, and the compound has good PDE9A kinase inhibition activity and drug-forming property, and can be applied to treatment of related diseases mediated by abnormal expression of PDE 9.

The technical scheme adopted by the invention is as follows:

scheme one, a PDE9 kinase (phosphodiesterase 9) inhibitor represented by the following general formula (I), or a pharmaceutically acceptable salt, solvate, polymorph and isomer thereof:

wherein n is selected from 0, 1 or 2;

x is selected from N or CR₁；

R₁Independently selected from hydrogen, C_1-6Alkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, C_3-6Cycloalkyl, halo C_1-6Alkyl, - (CH)₂)m_’-C_3-6Cycloalkyl, m' ═ 1 or 2;

R₂independently selected from hydrogen, C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkyl, halo C_1-6Alkoxy radical, C_3-6Cycloalkyl oxy, C_3-6Cycloalkylamino, halo C_3-6Cycloalkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, C_1-6Alkylsulfonyl radical, C_3-8Cycloalkylsulfonyl radical, C_1-6Alkylthio radical, C_2-8Alkenyl (C)_3-6) Cycloalkyl radical, C_2-8Alkynyl (C)_3-6) Cycloalkyl radical, C_1-6Alkylcarbonyl group, C_3-6Cycloalkyl carbonyl, - (CH)₂)_n’-3-14 membered cycloalkyl, - (CH)₂)_n’-5-14 membered aromatic ring, - (CH)₂)_n’-5-10 membered heterocyclyl, - (CH)₂)_n’-5-10 membered heteroaryl, said heterocyclyl, heteroaryl containing 1-3O, S and/or N atoms, wherein any ring atom sulfur of heterocyclyl, heteroaryl may be optionally oxidized to S (O) or S (O)₂Optionally having any ring member carbon substitutedOxidation to C (O), n' ═ 0, 1 or 2, cycloalkyl, heterocyclyl, aryl or heteroaryl rings optionally substituted with 1-3R₅Substitution;

wherein, when X is selected from CR₁And R is₁When it is hydrogen, R₂Can not be

R₃Independently selected from hydrogen, hydroxyl, amino, carboxyl, cyano, nitro, halogen atom, C_1-6Alkyl radical, C_3-6Cycloalkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyloxy, halo C_1-6Alkoxy, halo C_1-6Alkyl, halo C_3-6Cycloalkyl radical, C_2-8Alkenyl radical, C_2-8Alkynyl, amino C_1-6Alkyl radical, C_3-6Cycloalkylamino, C_1-6Alkylsulfonyl, 3-to 8-membered cycloalkylsulfonyl, C_1-6Alkylcarbonyl group, C_3-6Cycloalkyl carbonyl group, C_1-6Alkylthio, 3-14 membered cycloalkyl, 5-14 membered aromatic ring, -5-10 membered heterocyclyl, -5-10 membered heteroaryl, said heterocyclyl, heteroaryl containing 1-3O, S and/or N atoms, wherein any ring atom sulfur of heterocyclyl, heteroaryl may be optionally oxidized to S (O) or S (O)₂Optionally, any ring atom carbon may be oxidized to C (O), cycloalkyl, aryl, heteroaryl, heterocyclyl may be optionally substituted with 1-3R₆Substitution;

R₄independently selected from hydrogen, hydroxyl, amino, carboxyl, cyano, nitro, halogen atom, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyloxy, halo C_1-6Alkoxy, halo C_1-6Alkyl, halo C_3-6Cycloalkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, amino C_1-6Alkyl radical, C_3-6Cycloalkylamino, C_1-6Alkylsulfonyl, 3-to 8-membered cycloalkylsulfonyl, C_1-6Alkylcarbonyl or C_3-6Cycloalkyl carbonyl group, C_1-6Alkylthio or- (CH)₂)_n’-3-7 membered cycloalkyl, - (CH)₂)_n’-5-6 membered aromatic ring, - (CH)₂)_n’--5-7A heterocyclic radical, - (CH)₂)_n’-5-6 membered heteroaryl, said heterocyclyl, heteroaryl containing 1-3O, S and/or N atoms, wherein heterocyclyl, heteroaryl optionally any ring S may be optionally oxidized to S (O) or S (O)₂Optionally, any ring carbon may be oxidised to c (o), n' ═ 0, 1 or 2, cycloalkyl, aryl, heteroaryl and heterocyclyl groups optionally substituted with 1 to 3R₇Substitution;

R₅independently selected from hydrogen, hydroxyl, amino, carboxyl, cyano, nitro, halogen atom, trifluoromethyl, methoxy, - (CH)₂)_n’-3-14 membered cycloalkyl, - (CH)₂)_n’-5-14 membered aromatic ring, - (CH)₂)_n’- - (CH) 5-to 10-membered heterocyclic group₂)_n’-5-10 membered heteroaryl, said heterocyclyl, heteroaryl containing 0-3O, S and/or N atoms, wherein N' is 0, 1 or 2;

R₆、R₇independently selected from hydrogen, hydroxyl, amino, carboxyl, cyano, nitro, halogen atom, trifluoromethyl, C_1-6Alkyl radical, C_1-4Alkoxy radical, C_1-6Alkylsulfonyl radical, C_1-6Alkylsulfonyl radical C_1-6Alkyl radical, C_1-6Alkylsulfonyl radical C_1-6Alkoxy, aminosulfonylamino C_1-6Alkyl, methoxy, cyclopropyl, - (CH)₂)m_’-C_3-6Cycloalkyl, m' ═ 1 or 2.

A PDE9 kinase inhibitor according to scheme one, or a pharmaceutically acceptable salt, solvate, polymorph and isomer thereof, wherein the structural formula is shown as (II):

n is selected from 0, 1 or 2;

x is selected from N or CR₁；

R₁Independently selected from hydrogen, methyl, ethyl, cyclopropyl, -CH₂-cyclopropyl, trifluoromethyl;

ring A is selected from optional- (CH)₂)_n’-3-14 membered cycloalkyl, - (CH)₂)_n’-5-14 membered aromatic ring, - (CH)₂)_n’-5-10 membered heterocyclyl, - (CH)₂)_n’-5-10 membered heteroaryl, said heterocyclyl, heteroaryl containing 1-3O, S and/or N atoms, wherein ring a is optionally substituted with 1-3R₅By substitution, any ring atom sulfur may be optionally oxidized to S (O) or S (O)₂Any ring atom carbon may be optionally oxidized to c (o); n' is 0, 1 or 2;

wherein, when X is selected from CR₁And R is₁When it is hydrogen, R₂Can not be

R₅Independently selected from hydrogen, hydroxyl, amino, carboxyl, cyano, nitro, halogen atom, trifluoromethyl, methoxy, - (CH)₂)_n’-3-14 membered cycloalkyl, - (CH)₂)_n’-5-14 membered aromatic ring, - (CH)₂)_n’-5-10 membered heterocyclyl, - (CH)₂)_n’-5-10 membered heteroaryl, said heterocyclyl, heteroaryl containing 0-3O, S and/or N atoms, wherein N ═ 0, 1, or 2.

3. And the PDE9 kinase inhibitor according to the third scheme and the second scheme, or pharmaceutically acceptable salts, solvates, polymorphs and isomers thereof, wherein the structural general formulas are shown as (III-1) and (III-2):

wherein the content of the first and second substances,

ring A' is selected from a 5-7 membered cycloalkyl, a 5-7 membered aromatic ring, a 5-7 membered heterocyclyl or a 5-7 membered heteroaromatic ring, said heterocyclyl, heteroaryl containing 1-3O, S and/or N atoms, any ring atom sulphur being optionally oxidised to S (O) or S (O)₂Any ring atom carbon may be optionally oxidized to c (o);

ring A' is selected from 5-7 membered cycloalkyl, 5-7 membered aromatic ring, -5-7 membered heterocyclyl or-5-7 membered heteroaromatic ring, said heterocyclyl, heteroaryl containing 1-3O, S and/orThe N atom, sulfur being any ring atom, being optionally oxidized to S (O) or S (O)₂Any ring atom carbon may be optionally oxidized to c (o);

wherein ring A "cannot be

Ring A 'and ring A' are optionally substituted with 1-3R₅The substitution is carried out by the following steps,

R₅independently selected from hydrogen, hydroxyl, amino, carboxyl, cyano, nitro, halogen atoms, C_1-4Alkyl, cyclopropyl, trifluoromethyl, -CH₂And (3) cyclopropyl.

A PDE9 kinase inhibitor of scheme four, as set forth in scheme three, or a pharmaceutically acceptable salt, solvate, polymorph and isomer thereof, wherein,

ring A' is selected from 5-7 membered cycloalkyl, 5-7 membered heterocyclyl, said heterocyclyl containing 1-3O, S and/or N atoms, any ring atom sulfur optionally being oxidized to S (O) or S (O)₂Any ring atom carbon may be optionally oxidized to c (o);

ring A' is selected from 5-7 membered cycloalkyl, 5-7 membered heterocyclyl, said heterocyclyl containing 1-3O, S and/or N atoms, any ring atom sulfur optionally being oxidized to S (O) or S (O)₂Any ring atom carbon may be optionally oxidized to c (o);

wherein ring A "cannot be

Ring A 'and ring A' are optionally substituted with 1-3R₅The substitution is carried out by the following steps,

R₅independently selected from hydrogen, hydroxyl, amino, carboxyl, cyano, nitro, halogen atoms, C_1-4Alkyl, cyclopropyl, trifluoromethyl, -CH₂A cyclopropyl group;

R₃independently selected from hydrogen, 3-7 membered cycloalkyl, 5-7 membered aromatic ring, -5-7 membered heterocyclyl or-5-7 membered heteroaromatic ring, said heterocyclyl, heteroaryl containing 1-3O, S and/or N atoms, wherein cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with 1 atomOr 2R₆Optionally, sulfur S, any ring atom of the heterocyclyl, heteroaryl, may be oxidized to S (O) or S (O)₂Any ring atom carbon may be optionally oxidized to c (o);

R₆independently selected from hydrogen, hydroxyl, amino, carboxyl, cyano, nitro, halogen atoms, C_1-4Alkyl radical, C_1-4Alkoxy, cyclopropyl, trifluoromethyl, C_1-4Alkylsulfonyl radical, C_1-4Alkylsulfonyl radical C_1-4Alkyl radical, C_1-4Alkylsulfonyl radical C_1-4Alkoxy, aminosulfonylamino C_1-4Alkyl, -CH₂-cyclopropyl.

PDE9 kinase inhibitors according to scheme five, according to scheme four, or pharmaceutically acceptable salts, solvates, polymorphs and isomers thereof, wherein,

ring A' is selected from 5-7 membered heterocyclyl, said heterocyclyl containing 1-3O, S and/or N atoms, any ring atom sulfur being optionally oxidized to S (O) or S (O)₂Any ring atom carbon may be optionally oxidized to c (o);

ring A' is selected from 5-7 membered heterocyclyl containing 1-3O, S and/or N atoms, any ring atom sulfur being optionally oxidized to S (O) or S (O)₂Any ring atom carbon may be optionally oxidized to c (o);

wherein ring A "cannot be

Ring A 'and ring A' are optionally substituted with 1-3R₅The substitution is carried out by the following steps,

R₅independently selected from hydrogen, hydroxyl, amino, carboxyl, cyano, nitro, halogen atoms, C_1-4Alkyl, cyclopropyl, trifluoromethyl, -CH₂A cyclopropyl group;

R₄independently selected from hydrogen, hydroxyl, amino, carboxyl, cyano, nitro, halogen atoms, C_1-4Alkyl radical, C_1-4Alkoxy radical, C_3-6Cycloalkyloxy, halo C_1-4Alkoxy radical, C_3-6Cycloalkylamino, -CH₂-cyclopropyl, C_1-4Alkyl sulfonic acidAcyl, 3-8 membered cycloalkylsulfonyl, C_1-4Alkylcarbonyl or C_3-6A cycloalkyl carbonyl group.

A PDE9 kinase inhibitor according to scheme six, scheme five, or a pharmaceutically acceptable salt, solvate, polymorph and isomer thereof, wherein,

wherein the content of the first and second substances,

R₃preferred are the following groups: hydrogen, hydrogen,

Ring a' and ring a "are each independently selected from the following groups:

preferred compounds of the invention, pharmaceutically acceptable salts, solvate compounds, polymorphs and isomers thereof:

detailed description of the invention:

the "halogen" in the present invention means fluorine, chlorine, bromine, iodine, etc., and preferably fluorine atom, chlorine atom.

The term "oxo" as used herein means that any carbon atom in the substituent structure may be replaced by "-C (O) -; if containing heteroatoms, the heteroatoms may form oxides, e.g.

Can be covered

Alternatively, e.g. any ring S is optionally oxidized to S (O) or S (O)₂。

The term "halo" as used herein means that any one of the carbon atoms in a substituent may be substituted with one or more of the same or different halogens. "halogen" is as defined above.

Said "C" of the present invention_1-6The "alkyl group" means a straight chain or branched alkyl group derived from a hydrocarbon moiety having 1 to 6 carbon atoms by removing one hydrogen atom, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 1-methyl-2-methylpropyl, and the like. Said "C_1-4Alkyl "refers to the above examples containing 1 to 4 carbon atoms.

"C" according to the invention_1-6Alkylcarbonylamino group and C_1-6Alkylaminocarbonyl group and C_1-6Alkylsulfonyl "is independently C_1-6alkyl-C (O) -NH-, C_1-6alkyl-NH-C (O) -, C_1-6alkyl-S (O)₂-; said "C_1-6Alkyl "is as defined above, preferably" C_1-4Alkyl groups ".

"C" according to the invention_1-6Alkoxy "means" C "as defined hereinbefore_1-6Alkyl "a group attached to the parent molecular moiety through an oxygen atom, i.e." C_1-6alkyl-O- "groups, e.g. methoxy, ethoxy, propoxy, isopropoxy, n-propylButoxy, t-butoxy, n-pentoxy, neopentoxy, n-hexoxy, and the like. Said "C_1-4Alkoxy "refers to the above examples containing 1 to 4 carbon atoms, i.e." C_1-4An alkyl-O- "group.

The "cycloalkyl" groups described herein may be 3-14 membered cycloalkyl groups, including monocyclic cycloalkyl groups or fused ring cycloalkyl groups, and may be saturated, partially saturated or unsaturated, but not aromatic. Monocyclic cycloalkyl groups can be 3-8 membered cycloalkyl, 5-7 membered cycloalkyl, examples of which include, but are not limited to: cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclobutenyl, cyclopentenyl, cyclohexenyl, 1, 4-cyclohexadienyl, cycloheptenyl, 1, 4-cycloheptadienyl, cyclooctenyl, 1, 5-cyclooctadienyl, and the like. Fused ring cycloalkyl groups include fused ring cycloalkyl, bridged cycloalkyl, spirocycloalkyl groups, and may be saturated, partially saturated or unsaturated, but are not aromatic.

The fused cycloalkyl group may be a 6-12 membered fused cycloalkyl group, a 7-10 membered fused cycloalkyl group, examples of which include, but are not limited to: bicyclo [3.1.1] heptanyl, bicyclo [2.2.1] heptanyl, bicyclo [2.2.2] octanyl, bicyclo [3.2.2] nonanyl, bicyclo [3.3.1] nonanyl and bicyclo [4.2.1] nonanyl.

The spiro group may be 6-12-membered spiro ring group, 7-11-membered spiro ring group, and examples thereof include, but are not limited to:

the bridge ring group may be a 6-12-membered bridge ring group, a 7-11-membered bridge ring group, examples of which include, but are not limited to:

the term "heterocyclic group" as used herein means a "cycloalkyl group" in which any carbon atom may be substituted with a heteroatom selected from oxygen, sulfur and nitrogen, preferably 1 to 3 heteroatoms, and carbon, nitrogen and sulfur atoms may be substituted with oxo.

"heterocyclyl" means a 5-14 membered monocyclic heterocyclyl, bicyclic heterocyclyl system, or polycyclic heterocyclyl system, including saturated, partially saturated heterocyclyl groups, but excluding aromatic rings. "3-8" membered saturated heterocyclyl, examples of which include, but are not limited to, aziridinyl, oxetanyl, thietanyl, azetidinyl, oxetanyl, thietanyl, tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydrothienyl, imidazolidinyl, pyrazolidinyl, 1, 2-oxazolidinyl, 1, 3-oxazolidinyl, 1, 2-thiazolidinyl, 1, 3-thiazolidinyl, tetrahydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, piperidinyl, piperazinyl, morpholinyl, 1, 4-dioxanyl, 1, 4-oxathietanyl; "3-8" membered partially saturated heterocyclic group, examples of which include, but are not limited to, 4, 5-dihydroisoxazolyl, 4, 5-dihydrooxazolyl, 2, 3-dihydrooxazolyl, 3, 4-dihydro-2H-pyrrolyl, 2, 3-dihydro-1H-pyrrolyl, 2, 5-dihydro-1H-imidazolyl, 4, 5-dihydro-1H-pyrazolyl, 4, 5-dihydro-3H-pyrazolyl, 4, 5-dihydrothiazolyl, 2H-pyranyl, 4H-pyranyl, 2H-thiopyranyl, 4H-thiopyranyl, 2,3,4, 5-tetrahydropyridinyl, 1, 2-isooxazinyl, 1, 4-isooxazinyl or 6H-1, 3-oxazinyl and the like. Bicyclic heterocycles are bicyclic heterocyclic systems fused in a fused, spiro, bridged fashion to a phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocyclyl or monocyclic heteroaryl group. The heterocyclic group may be a 6-12 membered heterocyclic group, a 7-10 membered heterocyclic group, a 6-12 membered saturated heterocyclic group, a 7-8 membered saturated heterocyclic group, an 8 membered saturated heterocyclic group, examples of which include, but are not limited to: 3-azabicyclo [3.10 ] hexanyl, 3, 6-diazabicyclo [3.2.0] heptane, 3, 8-diazabicyclo [4.2.0] octanyl, 3, 7-diazabicyclo [4.2.0] octanyl, octahydropyrrolo [3,4-c ] pyrrole, octahydropyrrolo [3,4-b ] [1,4] oxazinyl, octahydro-1H-pyrrolo [3,4-c ] pyridine, 2, 3-dihydrobenzofuran-2-yl, 2, 3-dihydrobenzofuran-3-yl, indolin-1-yl, indolin-2-yl, indolin 3-yl, 2,3 dihydrobenzothien-2-yl, octahydro-1H-indolyl, substituted octahydro-1H-2-yl, 3-azabicyclo [3.2.0] octane, octahydropyrrolo [ 3.4.4 ] pyrrole [ 3.4-c ] pyridine, 2, 3-dihydrobenzofuran-2-yl, octahydrobenzofuranyl, octahydrocyclopenta [ c ] pyrrole, hexahydrocyclopenta [ c ] furan, 2-dioxohexahydrocyclopenta [ c ] thiophene.

The spiro heterocyclic group may be a 6-12 membered spiro heterocyclic group, a 7-11 membered spiro heterocyclic group, a 6-12 membered saturated spiro heterocyclic group, a 7 membered saturated spiro heterocyclic group, examples of which include, but are not limited to:

the bridged heterocyclic group may be 6-12 membered bridged heterocyclic group, 7-11 membered bridged heterocyclic group, 6-12 membered saturated bridged cyclic group, examples of 7-8 membered saturated bridged cyclic group include but are not limited to:

in certain embodiments, bicyclic heterocyclyl is a 5-or 6-membered monocyclic heterocyclyl ring fused to a phenyl ring, a 5-or 6-membered monocyclic cycloalkyl, a 5-or 6-membered monocyclic cycloalkenyl, a 5-or 6-membered monocyclic heterocyclyl or a 5-or 6-membered monocyclic heteroaryl, characterized in that the bicyclic heterocyclyl is optionally substituted with one or two groups which are independent oxo or thio groups.

"5-14 membered aryl" means a cyclic aromatic group containing 5 to 14 carbon atoms, including "6-8 membered monocyclic aryl", such as phenyl, cyclooctenyl, and the like; included are "8-14 membered fused ring aryl radicals" such as, for example, pentalene, naphthalene, phenanthrene, and the like. The term "aryl" as used herein refers to a phenyl group (i.e., a monocyclic aryl group) or a bicyclic ring system containing at least one benzene ring or only carbon atoms in an aromatic bicyclic ring system. The bicyclic aryl can be azulenyl, naphthyl, or phenyl fused to a monocyclic cycloalkyl, monocyclic cycloalkenyl, or monocyclic heterocycle. The bicyclic aryl is attached to the parent molecular species through any carbon atom contained in the phenyl portion of the bicyclic system or through any carbon atom bearing a naphthyl or azulene ring. The fused monocyclic cycloalkyl or monocyclic heterocyclyl portion of the bicyclic aryl is optionally substituted with one or two oxo and/or thia groups.

The term "heteroaryl" as used herein refers to a 5-14 membered monocyclic heteroaryl or bicyclic ring system containing at least one heteroaromatic ring. The monocyclic heteroaryl group may be a 5-or 6-membered ring. The 5-membered ring consists of two double bonds and one, two, three or four nitrogen atoms and one oxygen or sulfur atom. The 6-membered ring consists of three double bonds and one, two, three or four nitrogen atoms. The 5-or 6-membered heteroaryl is attached to the parent molecular species through any carbon or nitrogen atom contained within the heteroaryl. Representative examples of monocyclic heteroaryl groups include, but are not limited to, furyl, imidazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, oxazolyl, isoxazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyrrolyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazolyl, and triazinyl. Bicyclic heteroaryls consist of a monocyclic heteroaryl fused to a phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocyclyl or monocyclic heteroaryl. The cycloalkyl or heterocyclyl portion of the fused bicyclic heteroaryl is optionally substituted with one or two groups which are independently oxo or thio. When the bicyclic heteroaryl contains a fused cycloalkyl, cycloalkenyl, or heterocyclyl ring, then the bicyclic heteroaryl is attached to the parent molecular species through any carbon or nitrogen atom contained within the monocyclic heteroaryl portion of the bicyclic ring system. When the bicyclic heteroaryl is a monocyclic heteroaryl fused to a phenyl ring or a monocyclic heteroaryl, the bicyclic heteroaryl is attached to the parent molecular species through any carbon or nitrogen atom within the bicyclic system. Representative examples of bicyclic heteroaryls include, but are not limited to, benzimidazolyl, benzofuranyl, benzothienyl, benzooxadiazolyl, benzothiazolyl, cinnolinyl, 5, 6-dihydroquinolin-2-yl, 5, 6-dihydroisoquinolin-1-yl, indazolyl, indolyl, isoquinolinyl, naphthyridinyl, purinyl, quinolinyl, 5,6,7, 8-tetrahydroquinolin-2-yl, 5,6,7, 8-tetrahydroquinolinyl, 5,6,7, 8-tetrahydroquinolin-4-yl, 5,6,7, 8-tetrahydroisoquinolin-1-yl, 4,5,6, 7-tetrahydro [ c ] [1,2,5] oxadiazole and 6, 7-dihydro [ c ] [1,2,5] oxadiazol-4 (5H) -onyl. In certain embodiments, the fused bicyclic heteroaryl is a 5-or 6-membered monocyclic heteroaryl ring fused to a phenyl ring, a 5-or 6-membered monocyclic cycloalkyl, a 5-or 6-membered monocyclic cycloalkenyl, a 5-or 6-membered monocyclic heterocyclyl or a 5-or 6-membered monocyclic heteroaryl, wherein the fused cycloalkyl, cycloalkenyl and heterocyclyl are optionally substituted with one or two groups which are independent oxo or thio groups.

The invention also claims pharmaceutical formulations of any of the compounds of the invention or stereoisomers thereof or pharmaceutically acceptable salts, solvates, polymorphs and isomers thereof, characterised in that they contain one or more pharmaceutically acceptable carriers.

The pharmaceutical carrier of the present invention may be one or more solid or liquid filler or gel materials suitable for human use. The pharmaceutically acceptable carrier is preferably of sufficient purity and sufficiently low toxicity, and is compatible with the active ingredients of the present invention without significantly diminishing the efficacy of the active ingredient. For example, the pharmaceutically acceptable carrier may be a filler, a binder, a disintegrant, a lubricant, an aqueous solvent or a non-aqueous solvent, and the like.

The pharmaceutical preparation of the present invention may be prepared into any pharmaceutically acceptable dosage form, and administered to a patient or subject in need of such treatment by any suitable administration, for example, oral, parenteral, rectal, or pulmonary administration. For oral administration, it can be made into tablet, capsule, pill, granule, etc. For parenteral administration, it can be made into injection, injectable sterile powder, etc.

The present invention also claims a pharmaceutical composition of any one of the compounds of the present invention or stereoisomers or pharmaceutically acceptable salts, solvates, polymorphs, and isomers thereof, further comprising one or more second therapeutically active agents which are antimetabolites, growth factor inhibitors, inhibitors of the filamentation class, anti-tumor hormones, alkylating agents, metals, topoisomerase inhibitors, hormonal agents, immunomodulators, tumor suppressor genes, cancer vaccines, immune checkpoints, or antibodies and small molecule drugs associated with tumor immunotherapy.

The invention also claims the use of any one of the compounds of the invention or stereoisomers thereof or pharmaceutically acceptable salts, solvates, polymorphs and isomers thereof, for the manufacture of a medicament for the treatment of a disease associated with aberrant expression of PDE9, wherein the disease associated with aberrant expression of PDE9 is:

(a) for treatment of diseases accessible by inhibition of PDE 9;

(b) for treating a CNS disorder, said CNS disorder being: cognitive impairment associated with a disease or condition selected from perception, attention, cognition, learning or memory, cognitive impairment associated with alzheimer's disease or with alzheimer's disease, cognitive impairment associated with schizophrenia or with schizophrenia, cognitive impairment associated with depression or bipolar disorder, age-related memory loss, vascular dementia, craniocerebral trauma, stroke, dementia occurring after stroke, post-traumatic dementia, general attention impairment, childhood attention impairment with learning and memory problems, alzheimer's disease, lewy body dementia, frontotemporal dementia, corticobasal dementia, amyotrophic lateral sclerosis, huntington's disease, multiple sclerosis, thalamic degeneration, creutzfeldt-jacob dementia, HIV dementia, schizophrenia, koxsackia psychosis.

(c) For treating a disease or condition selected from: sleep disorders, bipolar disorders, metabolic syndrome, obesity, diabetes, hyperglycemia, dyslipidemia, impaired glucose tolerance or a disease of the testis, brain, small intestine, skeletal muscle, heart, lung, thymus or spleen, sickle cell disease.

The term "pharmaceutically acceptable salts" as used herein refers to pharmaceutically acceptable acid and base addition salts and solvates. Such pharmaceutically acceptable salts include salts of acids such as: hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, sulfurous acid, formic acid, toluenesulfonic acid, methanesulfonic acid, nitric acid, benzoic acid, citric acid, tartaric acid, maleic acid, hydroiodic acid, alkanoic acids (such as acetic acid, HOOC- (CH)₂)_n-COOH (wherein n is 0 to 4)), and the like. Non-toxic pharmaceutical base addition salts include salts of bases such as: sodium, sodium,Potassium, calcium, ammonium, and the like. The person skilled in the art is aware of a number of non-toxic pharmaceutically acceptable addition salts.

"isomers" of the compounds of formula (I) according to the invention are defined as enantiomers when asymmetric carbon atoms are present in the compounds of formula (I); when the compound has a carbon-carbon double bond or a cyclic structure, cis-trans isomers can be generated; tautomers can occur when ketones or oximes are present in the compounds, and all enantiomers, diastereomers, racemates, cis-trans isomers, tautomers, geometric isomers, epimers and mixtures thereof of the compounds of formula (I) are included within the scope of the present invention.

General notes on the structure presentation:

compounds with stereogenic centers: the structures depicted in the experimental section below do not necessarily show all stereochemical possibilities of the compounds, but only one. However, in these cases, terms such as "trans-racemic mixture" or "cis-racemic mixture" are added after the depicted structure to indicate other stereochemical choices.

Detailed Description

Example (b): synthesis of Compound 1:

step 1: synthesis of intermediate 1-1:

LiCl (8.45g,0.12mmol,0.1eq), acetonitrile (4.0L), 3-methylbutyraldehyde (99.14g,1.15mol,1.0eq) and diethoxyphosphonate ethyl formate (258.05g,1.15mol,1.0eq) were charged into a 5L four-necked flask, stirred at room temperature (28 ℃) for reaction for 15min, DBU (192.44g,1.27mol,1.1eq) was added, and stirred at room temperature (28 ℃) for reaction for 2 h. Concentrate, add saturated aqueous ammonium chloride (about 500mL), extract with EA (100 mL. times.3), combine the EA phases, dry over anhydrous magnesium sulfate, filter, and concentrate to give the crude product.

Step 2: synthesis of intermediates 1-2:

the crude product from the previous step was dissolved in THF (200mL), water (200mL) and lithium hydroxide monohydrate (144.76g,3.45mol,3.0eq) were added and heated to 70 ℃ for reaction overnight (10 h). Cooling to 0 deg.C, adjusting pH to about 1 with 1mol/L HCl, adding EA for extraction (100 mL. times.2), combining the organic phases, drying over anhydrous magnesium sulfate, filtering, and concentrating to obtain brown yellow oil (46.35g, yield: 32%)

And step 3: synthesis of intermediates 1 to 3:

the intermediate 1-2(25.00g,0.20mol,1.0eq), DCM (500mL) and DMF (1mL) were added to a 1L four-necked flask, oxalyl chloride (27.92g,0.22mol,1.1eq) was added dropwise at an ice bath temperature of 15 ℃ or lower, warmed to room temperature (25 ℃) for reaction for 8h, and concentrated to give a brown-yellow oil. In another 1L four-necked flask were charged (R) -4-phenyloxazolidin-2-one (32.64g,0.20mol,1.0eq), THF, LiCl (8.48g,0.20mol,1.0eq) and TEA (40.48g,0.40mol,2.0eq), stirred at room temperature (25 ℃) for 0.5h, cooled in an ice bath to 0 ℃, DMAP (2.44g,0.02mol,0.1eq) was added, THF (500mL) dissolved in the above tan oil was added dropwise, and the mixture was allowed to warm to room temperature (20-25 ℃) and reacted for 10 h. Adding saturated Na₂CO₃The solution (500mL) was separated, the aqueous phase was extracted with EA (150mL × 2), the organic phases were combined, washed with saturated aqueous NaCl (300mL × 5), dried over anhydrous magnesium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (PE: EA ═ 15:1) to give a white solid (12.78g, yield 23.4%).

And 4, step 4: synthesis of intermediates 1 to 4:

A1L four-necked flask was charged with the compound 1-3(12.78g,46.8mmol,1.0eq) and toluene (200mL), stirred with N- (methoxymethyl) -N- (trimethylsilylmethyl) benzylamine (1.32g,56.1mmol,1.2eq) at room temperature (25 ℃ C.) for 0.5h, cooled to 0 ℃ in an ice bath, and a mixed solution of TFA (1mL) and DCM (8mL) was added dropwise and allowed to slowly warm to room temperature (25 ℃ C.) for 8 h. Saturated aqueous sodium bicarbonate (300mL) was added, the solution was separated, the toluene phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and the crude product was subjected to silica gel column chromatography (PE: EA 15:1-5:1), then PE (100mL) was added and slurried, filtered, and dried to obtain a white solid (10.89g, yield: 57.3%).

And 5: synthesis of intermediates 1 to 5:

a1000 mL two-necked flask was charged with intermediates 1-4(50.0g,123.0mmol,1.0eq) and THF (459mL), cooled to 0 deg.C in an ice bath, and charged with lithium hydroxide monohydrate (12.9g,307.5mmol,2.5eq), H₂O₂(34.9g,307.5mmol,2.5eq) and H₂O, reacting at 0 ℃ for 2 h. Adding Na₂S₂O₃(3.0eq), stirred for 0.5h, separated, extracted with aqueous phase EA (50 mL. times.2), and added NaH under ice bath₂PO₄(3.3eq), adjusted to pH 4-5 with 6mol/L HCl, extracted with DCM and i-PrOH (3:1) (200 mL. times.6), dried over anhydrous magnesium sulfate, filtered, and concentrated to a yellow oil (31g, yield: 96.87%).

Step 6: synthesis of intermediates 1 to 6:

to a 250mL single neck flask were added intermediates 1-5(30.45g,116.5mmol,1.0eq), THF (300mL), MeCN (300mL), DIPEA (30.1g,233.0mmol,2.0eq) and HATU (48.75g,128.2mmol,1.1eq), stirred at room temperature (25 ℃) for 5H, 1-amino-1H-pyrrole-2-carboxamide hydrochloride (18.81g,116.5mmol,1.0eq) was added, stirred at room temperature (25 ℃) for 8H, concentrated, EA (500mL) was added, the layers were separated, the organic phase was washed with saturated aqueous NaCl (150mL × 5), dried over anhydrous magnesium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (DCM: MeOH ═ 50:1-20:1) to give a pale yellow solid (27.0g, yield: 64.3%).

And 7: synthesis of intermediates 1 to 7:

a100 mL single neck flask was charged with intermediates 1-6(24.00g,65.0mmol,1.0eq) and EtOH (360mL), dissolved with stirring, added with KOH (10.92g,195.0mmol,3.0eq) and water, and heated to vigorous reflux for 20 h. After concentration, ice water (300mL) was added, DCM extraction (300mL × 2) was performed, the organic phases were combined, dried over anhydrous magnesium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (DCM: MeOH ═ 50:1) to give a pale yellow oil (10.0g, yield: 44.1%).

And 8: synthesis of intermediates 1 to 8:

a100 mL single vial was charged with intermediates 1-7(10.0g,28.5mmol,1.0eq), DCM (110mL) and TFA (55mL), dried in a tube, cooled to 0 deg.C in an ice bath, NBS (4.57g,25.7mmol,0.9eq) added in portions, incubated for 2h, warmed to room temperature (20 deg.C) for 8h, and LCMS checked for no material remaining. Concentrate, dissolve in DCM and saturate with Na₂CO₃The aqueous solution (200mL) was washed, the organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (DCM: MeOH: 400:1) to give a pale yellow solid (7.0g, yield: 57.4%).

And step 9: synthesis of intermediates 1 to 9:

intermediate 1-8(0.20g,0.466mmol) was dissolved in toluene (4mL) and morphine (4) was added8.7mg,0.559mmol)、Pd₂(dba)₃(8.6mg,0.009mmol), sodium tert-butoxide (62.9mg,0.652mmol), BINAP (11.9mg,0.019 mmol). N is a radical of₂Heating to 110 ℃ under protection, and refluxing for 8 h. The reaction was monitored by TLC, the reaction was concentrated, water (10mL) was added, dichloromethane was extracted (3 × 25mL), the organic phases were combined, washed successively with water (2 × 10mL), saturated brine (1 × 20mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was chromatographed over silica gel (eluted successively with DCM: MeOH ═ 200:1, DCM: MeOH ═ 150:1, DCM: MeOH ═ 100:1) to give a yellow solid (170mg, yield: 84.2%).

Step 10: synthesis of intermediates 1 to 10:

intermediate 1-9(170mg,0.39mmol,1.0eq) was dissolved in MeOH (5mL), Pd/C (cat.) was added to replace the hydrogen three times and the reaction was allowed to proceed overnight at room temperature (20 ℃). Filtration was carried out, the filter cake was washed with a small amount of methanol, and the filtrate was concentrated to give a yellow product (134.73mg, yield: 100.0%).

Step 11: synthesis of Compound 1:

intermediate 1-10(134.73mg,0.39mmol) was dissolved in DCM (6mL), 4-carboxaldehyde-1-methylpyridin-2 (1H) -one (64.2mg,0.468mmol) and NaBH (OAc) were added₃(206.6mg,0.975mmol), and stirred at room temperature overnight. TLC monitored the reaction for completion, saturated aqueous sodium bicarbonate (5mL) was added, extracted with DCM (3X 10mL), the organic phases combined, washed with water (2X 5mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by preparative HPLC to give a white solid (10.0mg, yield: 5.5%).

¹HNMR(400MHz,CDCl₃)δ(ppm):9.96(s,1H),7.36-7.34(d,1H),6.99-6.98(d,1H),6.45(s,1H),6.41-6.40(d,1H),5.96-5.95(d,1H),3.92(s,4H),3.60-3.57(d,1H),3.53(s,2H),3.40-3.29(m,4H),3.18-3.15(t,2H),2.99-2.96(d,1H),2.48-2.44(m,3H),1.95-1.90(t,1H),1.64-1.62(d,1H),1.41-1.34(m,2H),1.29-1.26(m,1H),0.92-0.88(m,6H).

Molecular formula C₂₅H₃₄N₆O₃Molecular weight 466.59 LC-MS (M/z) ═ 467.28[ M + H⁺].

Example (b): synthesis of Compound 2:

step 1: preparation of intermediate 2-1:

compound 1(47.0g,0.55mol,1.0eq), DCM (186.0mL), MeCN (940.0mL), and TEMPO (4.30g,0.03mol,0.05eq) were added to a 3L four-necked flask, iodobenzene diacetate (186.0g,0.58mol,1.05eq) was added in portions, and the reaction was carried out at room temperature (10-15 ℃ C.) for 8 h. Saturated aqueous sodium bicarbonate (1.0L) and DCM (1.0L) were added, the layers separated, the aqueous phase was extracted with DCM (200 mL. times.1), the organic phases combined, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure below 30 ℃ to give about 600mL of the product as a MeCN solution in 100% yield.

Step 2: synthesis of intermediate 2-2:

LiCl (34.1g,0.83mmol,1.5eq), acetonitrile (1.5L), diethoxyphosphonyl ethyl formate (147.9g,0.66mol,1.2eq) and DBU (126.3g,0.66mol,1.2eq) were added to a 3L four-necked flask, stirred for 1h, cooled to 0 ℃ in an ice bath, added dropwise to the solution of the prepared intermediate 2-1 (600mL,0.55mol,1.0eq) and slowly warmed to room temperature (10-15 ℃) for reaction for 8 h. After concentration, EA (500mL) was added, and a cooled aqueous sodium bicarbonate solution (1.0L) was added to the residue, followed by liquid separation, washing of the EA phase with a saturated aqueous NaCl solution, drying over anhydrous magnesium sulfate, filtration, and purification of the concentrated crude product by silica gel column chromatography (EA: PE ═ 0-1:20) to obtain a colorless transparent oil (43.8g, yield: 51.7%).

And step 3: synthesis of intermediates 2 to 3:

A1.0L four-necked flask was charged with lithium hydroxide monohydrate (35.8g,0.85mol,3.0eq) dissolved in intermediate 2-2(43.8g,0.28mol,1.0eq), THF (200.0mL) and water (200.0mL), and heated to reflux (70 ℃ C.) for 8 h. The temperature was lowered to 0 ℃ in an ice bath, the pH was adjusted to about 1 with 6mol/L hydrochloric acid, the mixture was separated, aqueous phase EA was extracted (50 mL. times.2), organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated to give a pale yellow oil (30.9g, yield: 87.3%).

And 4, step 4: synthesis of intermediates 2 to 4:

the intermediate 2-3(20.0g,0.16mol,1.0eq), DCM (400mL) and DMF (1mL) were added to a 1L four-necked flask, oxalyl chloride (22.1g,0.17mol,1.1eq) was added dropwise at an ice bath temperature of 15 ℃ or lower, warmed to room temperature (10 ℃) for reaction for 3h, and concentrated to give a brown-yellow oil. In another 1L four-necked flask were charged (R) -4-phenyloxazolidin-2-one (27.7g,0.17mol), THF (200mL), LiCl (7.78g,0.18mol,1.1eq) and TEA (32.38g,0.32mol,2.0eq), stirred at room temperature (10 ℃ C.) for 2h, cooled to 0 ℃ in an ice bath, DMAP (1.95g,0.016mol,0.1eq) was added dropwise thereto, and the mixture was allowed to spontaneously warm to room temperature (0-10 ℃ C.) to react. Adding saturated Na₂CO₃The solution (300mL) was separated, the aqueous phase was extracted with EA (100mL × 2), the organic phases were combined, dried over anhydrous magnesium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (PE: EA ═ 6:1) to give a white solid (21.5g, yield 50.0%).

And 5: synthesis of intermediates 2 to 5:

in a 1.0L four-necked flask, intermediate 2-4(21.5g,79.2mmol,1.0eq), toluene (200.0mL) and the compound N- (methoxymethyl) -N- (trimethylsilylmethyl) benzylamine (20.2g,87.2mmol,1.1eq) were added, stirred for 1h, cooled to 0 ℃ in an ice bath, TFA (0.7mL) and DCM (5.6mL) were slowly added dropwise, slowly warmed to room temperature (15 ℃) and reacted for 10h, a saturated aqueous sodium bicarbonate solution (300mL) was added, the solution was separated, the organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (PE: EA ═ 10:1-6:1) and then slurried with PE to give a white mass (13.00g, yield: 40.6%).

Step 6: synthesis of intermediates 2 to 6:

the intermediate 2-5(11.0g,27.2mmol,1.0eq), THF (200.0mL) was added to a 1L four-necked flask, cooled to 0 ℃ in an ice bath and H was added dropwise₂O₂(6.17g,54.4mol,2.0eq) in water (25mL), stirred for 0.5h, and reacted at 0 ℃ for 1.5h with lithium hydroxide monohydrate (2.85g,68.0mmol,2.5eq) in water (25mL) added dropwise at 0 ℃. Adding Na₂S₂O₃(3.0eq), Water (50.0L), stirring at room temperature for 1h, adding EA (200.0mL), separating, extracting with aqueous EA (100.0 mL. times.1), cooling to 0 deg.C in ice bath, adding NaH₂PO₄(3.3eq), supplement with Na₂S₂O₃(3.0 eq.) and reacted at room temperature (15 ℃ C.) for 8 hours, extracted with DCM (100 mL. times.8), dried over anhydrous magnesium sulfate, filtered, and concentrated to give an off-white solid (7.10g, yield: 100%).

And 7: synthesis of intermediates 2 to 7:

in a 500.0mL single neck flask were added intermediates 2-6(6.00g,23.13mmol,1.0eq), THF (60.0mL), MeCN (120.0mL) and HATU (9.67g,25.44mmol,1.1eq) and reacted at room temperature (15-20 ℃ C.) for 5H, followed by addition of the compound 1-amino-1H-pyrrole-2-carboxamide hydrochloride (3.74g,23.13mmol,1.0eq) and stirring at room temperature (10-15 ℃ C.) for 10H. Concentrated, EA (250.0mL), washed with saturated aqueous sodium chloride (200.0mL × 5), dried over anhydrous magnesium sulfate, filtered, concentrated, and the crude product purified by silica gel column chromatography (successively with DCM: MeOH ═ 150:1, DCM: MeOH ═ 100:1, DCM: MeOH ═ 70:1, DCM: MeOH ═ 30:1) to give a pale yellow powder (3.80g, yield: 44.7%).

And 8: synthesis of intermediates 2 to 8:

a100 mL single neck flask was charged with intermediate 2-7(1.00g,2.73mmol,1.0eq) and EtOH (10mL), dissolved with stirring, and heated to vigorous reflux for 10h with aqueous KOH (0.23g,4.1mmol,1.5eq) (10 mL). Concentrating, adding methanol (20mL), concentrating, adding THF (20.0mL), performing ultrasonic treatment for 5min, adding a small amount of anhydrous magnesium sulfate, drying, filtering, washing a filter cake with a small amount of THF, concentrating the filtrate, and purifying the crude product by silica gel column chromatography (PE: EA ═ 2:1-1:1) to obtain a white solid (0.42g, yield: 44.0%).

And step 9: synthesis of intermediates 2 to 9:

a100 mL single vial was charged with intermediate 2-8(0.42g,1.21mmol,1.0eq), DCM (15mL) and TFA (8.0mL) were added under ice-bath, stirring was carried out for 0.5h, the ice-bath was cooled to 0 deg.C, NBS (0.19g,1.08mmol,0.9eq) was added in portions, about 1h addition was completed, reaction was carried out at room temperature (15 deg.C) for 8h, LCMS detected that about 15% of the starting material remained, cooling was carried out to 0 deg.C, 0.1eq of NBS was added, and reaction was carried out at room temperature (15 deg.C) for 2 h. After concentration, DCM (50.0mL) and a saturated aqueous solution of sodium bicarbonate (20.0mL) were added, the mixture was stirred for 5min, the mixture was separated, aqueous DCM was extracted (50mL × 1), the organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and the crude product was concentrated and purified by silica gel column chromatography (PE: EA ═ 4:1-2:1) to obtain an off-white solid (400.0mg, yield: 77.3%).

Step 10: synthesis of intermediates 2 to 10:

intermediate 2-9(0.20g,0.468mmol) was dissolved in toluene (4mL) and morpholine (48.9mg,0.5616mmol) and Pd were added₂(dba)₃(9.0mg,0.0094mmol), sodium tert-butoxide (63.2mg,0.655mmol) and BINAP (11.7mg,0.019mmol), N₂Heating to 110 ℃ under protection, and refluxing for 8 h. The reaction was monitored by TLC, the reaction was concentrated, water (10mL) was added, and the mixture was extracted with dichloromethane (3 × 25mL), washed with water (2 × 10mL), washed with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was chromatographed on a silica gel column (eluting with DCM: MeOH ═ 200:1, DCM: MeOH ═ 150:1, DCM: MeOH ═ 100:1 in this order) to give a yellow solid (80mg, yield: 39.6%).

Step 11: synthesis of intermediates 2 to 11:

intermediate 2-10(100mg,0.23mmol 1.0eq) was dissolved in MeOH (4mL) and DCM (20mL), Pd/C (cat.) was added to replace the hydrogen three times and the reaction was allowed to proceed overnight at room temperature (20 ℃). Filtration was carried out, the filter cake was washed with a small amount of methanol, and the filtrate was concentrated to give a yellow product (30.0mg, yield: 37.5%).

Step 12: synthesis of Compound 2:

intermediate 2-11(30mg,0.087mmol) was dissolved in DCM (2mL) and 4-formyl-1-methylpyridin-2 (1H) -one (14.3mg,0.104mmol), NaBH (OAc) was added₃(46.1mg,0.218 mmol). Stir at room temperature overnight. The reaction was monitored by TLC, saturated aqueous sodium bicarbonate (5mL) was added, extracted with DCM (3 × 10mL), the organic phases were combined, washed with water (2 × 5mL), dried over anhydrous sodium sulfate, filtered, concentrated and the crude was purified by silica gel column chromatography (DCM: MeOH ═ 200:1-20:1) to give a white solid (5.8mg, yield: 14.5%).

¹HNMR(400MHz,CDCl₃)δ(ppm):9.92(s,1H),7.36-7.34(d,1H),6.99-6.98(d,1H),6.45(s,1H),6.42-6.40(d,1H),5.96-5.94(d,1H),3.92-3.89(m,4H),3.61-3.58(d,1H),3.53(s,2H),3.42-3.37(m,2H),3.32-3.27(m,2H),3.19-3.14(m,2H),3.00-2.93(m,3H),2.48-2.44(m,2H),2.03-1.99(t,2H),1.53(s,2H),1.27(s,4H).

Molecular formula C₂₅H₃₂N₆O₃Molecular weight 464.57 LC-MS (M/z) ═ 465.3[ M + H⁺].

Example 37 Synthesis of- (1, 1-dioxotetrahydro-2H-thiopyran-4-yl) -2- ((3S,4S) -4-isobutyl-1- ((1-methyl-2-oxo-1, 2-dihydropyridin-4-yl) methyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (Compound 10)

The method comprises the following steps:

step 1 Synthesis of 2- ((3S,4S) -1-benzyl-4-isobutylpyrrolidin-3-yl) -7- (3, 6-dihydro-2H-thiopyran-4-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

reacting 2- (3S,4S) -1-benzyl-4-isobutyl pyrrolidine-3-yl) -7-bromo pyrrolo [2,1-f][1,2,4]Triazine-4 (3H) -one (1.0g,2.33mmol) was dissolved in 1, 4-dioxane (30mL), and 2- (3, 6-dihydro-2H-thiopyran-4-yl) -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolane (632.3mg,2.796mmol), potassium carbonate (644.1mg,4.66mmol), Pd (dppf) Cl, was added₂(2.05g mg,2.796mmol) and water (15 mL). Heating to 100 deg.C under nitrogen protection, reflux reacting for 8 hr, monitoring by TLC, concentrating the reaction solution, dissolving in dichloromethane (150mL), filtering, concentrating the filtrate, and purifying with silica gelPerforming column chromatography (DCM: MeOH: 400:1-100:1) to obtain the product 2- ((3S,4S) -1-benzyl-4-isobutyl pyrrolidine-3-yl) -7- (3, 6-dihydro-2H-thiopyran-4-yl) pyrrolo [2, 1-f)][1,2,4]Triazin-4 (3H) -one (580mg, yield: 55.5%).

Step 2 Synthesis of 2- ((3S,4S) -1-benzyl-4-isobutylpyrrol-3-yl) -7- (1, 1-dioxo-3, 6-dihydro-2H-thiopyran-4-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

2- ((3S,4S) -1-benzyl-4-isobutylpyrrolidin-3-yl) -7- (3, 6-dihydro-2H-thiopyran-4-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (200.0mg,0.446mmol) was dissolved in methanol (10mL), the ice salt bath was cooled to 0 ℃, a solution of Oxone (1.096g) in water (6mL) was added, and the mixture was stirred for 2 hours under the ice salt bath. After TLC to monitor the reaction was complete, water (60mL) was added under ice salt bath, extracted with ethyl acetate (3X 40mL), the organic phases combined, dried over anhydrous magnesium sulfate, filtered and concentrated to a white solid (100.0mg crude).

Step 3 Synthesis of 7- (1, 1-dioxotetrahydro-2H-thiopyran-4-yl) -2- ((3S,4S) -4-isobutylpyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

2- ((3S,4S) -1-benzyl-4-isobutylpyrrol-3-yl) -7- (1, 1-dioxo-3, 6-dihydro-2H-thiopyran-4-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (100.0mg crude) was dissolved in methanol (5mL), Pd/C (5.0mg) was added, hydrogenation was carried out for 6 hours, and completion of the reaction was monitored by TLC. Suction filtration was carried out on celite, and the filtrate was concentrated under reduced pressure to give 7- (1, 1-dioxotetrahydro-2H-thiopyran-4-yl) -2- ((3S,4S) -4-isobutylpyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (88.0mg).

Step 4 Synthesis of 7- (1, 1-dioxotetrahydro-2H-thiopyran-4-yl) -2- ((3S,4S) -4-isobutyl-1- ((1-methyl-2-oxo-1, 2-dihydropyridin-4-yl) methyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

7- (1, 1-dioxotetrahydro-2H-thiopyran-4-yl) -2- ((3S,4S) -4-isobutylpyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (81.64mg) was dissolved in DCM (4mL), 4-carboxaldehyde-1-methylpyridin-2 (1H) -one (34.23mg,0.25mmol) and sodium triacetoxyborohydride (110.2mg,0.52mmol) were added and stirred at room temperature overnight. TLC to monitor the reaction was complete, saturated aqueous sodium bicarbonate (5mL) was added, extracted with DCM (3 × 10mL), combined with organic phase, washed with water (2 × 5mL), dried over anhydrous sodium sulfate, filtered, concentrated and the crude product was separated by preparative thin layer chromatography (DCM: MeOH ═ 10:1) to give 7- (1, 1-dioxotetrahydro-2H-thiopyran-4-yl) -2- ((3S,4S) -4-isobutyl-1- ((1-methyl-2-oxo-1, 2-dihydropyridin-4-yl) methyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one as a white solid (4.0mg, yield: 3.7%).

¹HNMR(400MHz,DMSO-d₆)δ(ppm):11.44(s,1H),7.61-7.59(d,1H),6.81-6.80(d,1H),6.42-6.41(d,1H),6.33(s,1H),6.16-6.14(d,1H),3.45-3.43(d,3H),3.37(s,5H),3.15-3.09(t,3H),2.92-2.90(d,2H),2.82(s,2H),2.71-2.66(d,2H),2.32-2.21(m,5H),2.17-2.09(m,2H),1.55-1.50(m,2H),1.48-1.31(m,3H),0.87-0.83(m,6H).

Molecular formula C₂₆H₃₅N₅O₄Molecular weight of S513.66 LC-MS (Pos, M/z) ═ 514.5[ M + H⁺].

Example 47 Synthesis of- (1, 1-thiomorpholinyl) -2- ((3S,4S) -4-isobutyl-1- ((1-methyl-2-oxo-1, 2-dihydropyridin-4-yl) methyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (Compound 15)

The method comprises the following steps:

step 1 Synthesis of 2- ((3S,4S) -1-benzyl-4-isobutylpyrrol-3-yl) -7- (1, 1-thiomorpholinyl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

reacting 2- (3S,4S) -1-benzyl-4-isobutyl pyrrolidine-3-yl) -7-bromo pyrrolo [2,1-f][1,2,4]Triazin-4 (3H) -one (1.0g,2.33mmol) was dissolved in toluene (20mL), thiomorpholine 1, 1-dioxide hydrochloride (956.2mg,5.592mmol), Pd was added₂(dba)₃(88.0mg,0.093mmol), sodium tert-butoxide (1.35g,13.98mmol) and BINAP (116.3mg,0.186mmol), N₂The reaction was heated to 110 ℃ under reflux overnight with protection. The reaction was monitored by TLC, the reaction mixture was concentrated, water (10mL) was added, dichloromethane was extracted (3 × 25mL), the organic phases were combined, washed with water (2 × 10mL), washed with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was chromatographed on silica gel column (DCM: MeOH ═ 200:1-10:1) to give the product (380mg, yield: 33.8%).

Step 2 Synthesis of (3S,4S) -3- (7- (1, 1-thiomorpholino) -4-oxo-3, 4-dihydropyrrolo [2,1-f ] [1,2,4] triazin-2-yl) -4-isobutylpyrrolidine-1-carboxylic acid tert-butyl ester:

dissolving 2- ((3S,4S) -1-benzyl-4-isobutylpyrrol-3-yl) -7- (1, 1-thiomorpholinyl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (380.0mg,0.786mmoL) in dichloromethane (1.5mL) and methanol (1.5mL), adding di-tert-butyl dicarbonate (171.63mg,0.786mmoL) and Pd/C (19.0mg), replacing hydrogen three times, hydrogenating overnight, monitoring the reaction by TLC for completion, filtering with diatomaceous earth, and concentrating the filtrate under reduced pressure to obtain (3S,4S) -3- (7- (1, 1-thiomorpholinyl) -4-oxo-3, 4-dihydropyrrolo [2,1-f ] [1,2,4] triazin-2-yl) -4-isobutylpyrrolidin-1- Tert-butyl formate (390.0mg of crude product).

Step 3 Synthesis of 7- (1, 1-thiomorpholinyl) -2- ((3S,4S) -4-isobutylpyrrol-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

tert-butyl (3S,4S) -3- (7- (1, 1-thiomorpholino) -4-oxo-3, 4-dihydropyrrolo [2,1-f ] [1,2,4] triazin-2-yl) -4-isobutylpyrrolidine-1-carboxylate (390.0mg crude) was dissolved in dichloromethane (5mL), trifluoroacetic acid (2.5mL) was added dropwise at 0 deg.C, the reaction was slowly warmed to room temperature for 2 hours, and the completion of the reaction was monitored by TLC. The reaction solution was concentrated under reduced pressure to give 7- (1, 1-thiomorpholinyl) -2- ((3S,4S) -4-isobutylpyrrol-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (390.0mg crude).

Step 4 Synthesis of 7- (1, 1-thiomorpholinyl) -2- ((3S,4S) -4-isobutyl-1- ((1-methyl-2-oxo-1, 2-dihydropyridin-4-yl) methyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

7- (1, 1-thiomorpholinyl) -2- ((3S,4S) -4-isobutylpyrrol-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (309.1mg) was dissolved in DCM (6mL), 4-aldehyde-1-methylpyridin-2 (1H) -one (128.9mg,0.94mmol) and sodium triacetoxyborohydride (416.5mg,1.965mmol) were added, stirring was carried out overnight at room temperature, the reaction was monitored by TLC for completion, a saturated aqueous sodium bicarbonate solution (5mL) was added, extraction was carried out with DCM (3X 10mL), organic phase was combined, water washing was carried out (2X 5mL), dried over anhydrous sodium sulfate, filtration and concentration were carried out, and the crude product was purified by silica gel column chromatography (DCM: MeOH ═ 100:1-20:1) to give 7- (1) as a yellow solid, 1-Sulfur dioxide morpholino) -2- ((3S,4S) -4-isobutyl-1- ((1-methyl-2-oxo-1, 2-dihydropyridin-4-yl) methyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (80.0mg, yield: 19.8%)

¹HNMR(400MHz,DMSO-d₆)δ(ppm):7.61-7.59(d,1H),6.79-6.78(d,1H),6.31(s,1H),6.17-6.15(m,2H),3.67-3.66(d,4H),3.44-3.43(d,2H),3.38(s,3H),3.29-3.28(d,4H),2.94-2.85(m,2H),2.82-2.80(d,1H),2.78-2.67(m,2H),2.29-2.26(m,1H),1.54-1.50(m,2H),1.38-1.33(m,2H),0.87-0.83(m,6H).

Molecular formula C₂₅H₃₄N₆O₄Molecular weight of S514.65 LC-MS (Pos, M/z) ═ 515.2[ M + H⁺].

Example Synthesis of- ((3S,4S) -4- (cyclopropylmethyl) -1- ((1-methyl-2-oxo-1, 2-dihydropyridin-4-yl) methyl) pyrrolidin-3-yl) -7- (1, 1-thiomorpholinyl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (Compound 16)

The method comprises the following steps:

step 1 Synthesis of 2- ((3S,4S) -1-benzyl-4- (cyclopropylmethyl) pyrrolidin-3-yl) -7- (1, 1-thiomorpholinyl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

2- ((3S,4S) -1-benzyl-4- (cyclopropylmethyl) pyrrolidin-3-yl) -7-bromopyrrolo [2,1-f][1,2,4]Triazin-4 (3H) -one (200.0mg,0.468mmol) was dissolved in toluene (5mL), thiomorpholine 1, 1-dioxide hydrochloride (192.2mg,1.124mmol), Pd were added₂(dba)₃(17.86mg,0.0188mmol), sodium tert-butoxide (270.8mg,2.808mmol) and BINAP (23.76mg,0.038mmol) were heated to 110 ℃ under nitrogen and the reaction refluxed for 8 hours. TLC to monitor the reaction completion, the reaction was concentrated, water (10mL) was added, extraction was performed with dichloromethane (3X 25mL), the organic phases were combined, washed with water (2X 10mL), washed with saturated brine (1X 20mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was subjected to silica gel column chromatography (DCM: MeOH ═ 200:1-100:1)Obtaining the product 2- ((3S,4S) -1-benzyl-4- (cyclopropylmethyl) pyrrolidine-3-yl) -7- (1, 1-sulfur dioxide morpholine) pyrrolo [2, 1-f)][1,2,4]Triazin-4 (3H) -one (160mg, yield: 71.1%).

Step 2 Synthesis of tert-butyl (3S,4S) -3- (cyclopropylmethyl) -4- (7- (1, 1-dioxothiomorpholino) -4-oxo-3, 4-dihydropyrrolo [2,1-f ] [1,2,4] triazin-2-yl) pyrrolidine-1-carboxylate:

dissolving 2- ((3S,4S) -1-benzyl-4- (cyclopropylmethyl) pyrrolidin-3-yl) -7- (1, 1-thiomorpholinyl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (160.0mg,0.332mmoL) in dichloromethane (3.0mL) and methanol (3.0mL), adding di-tert-butyl dicarbonate (73.6mg,0.332mmoL) and Pd/C (8.0mg), displacing hydrogen three times, hydrogenating overnight, TLC monitoring completion of the reaction, suction-filtering with celite, concentrating the filtrate under reduced pressure to give (3S,4S) -3- (cyclopropylmethyl) -4- (7- (1, 1-dioxothiomorpholinyl) -4-oxo-3, 4-dihydropyrrolo [ 2), 1-f ] [1,2,4] triazin-2-yl) pyrrolidine-1-carboxylic acid tert-butyl ester (165.3mg crude).

Step 3 Synthesis of 2- ((3S,4S) -4- (cyclopropylmethyl) pyrrolidin-3-yl) -7- (1, 1-thiomorpholino) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

tert-butyl (3S,4S) -3- (cyclopropylmethyl) -4- (7- (1, 1-dioxothiomorpholino) -4-oxo-3, 4-dihydropyrrolo [2,1-f ] [1,2,4] triazin-2-yl) pyrrolidine-1-carboxylate (162.1mg crude) was dissolved in dichloromethane (4mL), trifluoroacetic acid (2mL) was added dropwise at 0 ℃ and allowed to slowly warm to room temperature for 2 hours, TLC monitored for completion of the reaction, the reaction mixture was concentrated under reduced pressure to give 2- ((3S,4S) -4- (cyclopropylmethyl) pyrrolidin-3-yl) -7- (1, 1-thiomorpholino) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (130.5mg, crude).

Step 4 Synthesis of 2- ((3S,4S) -4- (cyclopropylmethyl) -1- ((1-methyl-2-oxo-1, 2-dihydropyridin-4-yl) methyl) pyrrolidin-3-yl) -7- (1, 1-thiomorpholinyl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

2- ((3S,4S) -4- (cyclopropylmethyl) pyrrolidin-3-yl) -7- (1, 1-thiomorpholino) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (129.98mg) was dissolved in DCM (6mL), and 4-carboxaldehyde-1-methylpyridin-2 (1H) -one (54.6mg,0.398mmol) and sodium triacetoxyborohydride (254.3mg,1.2mmol) were added and stirred at room temperature overnight. The reaction was monitored by TLC for completion, saturated aqueous sodium bicarbonate (5mL) was added, extracted with DCM (3 × 10mL), the organic phases combined, washed with water (2 × 5mL), dried over anhydrous sodium sulfate, filtered, concentrated and the crude product purified by silica gel column chromatography (DCM: MeOH ═ 100:1-20:1) to give 2- ((3S,4S) -4- (cyclopropylmethyl) -1- ((1-methyl-2-oxo-1, 2-dihydropyridin-4-yl) methyl) pyrrolidin-3-yl) -7- (1, 1-thiomorpholinyl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one as a white solid (39.5mg, yield: 23.2%).

¹HNMR(400MHz,CDCl₃)δ(ppm):7.35-7.33(d,1H),6.97-6.96(d,1H),6.46(s,1H),6.39-6.37(d,1H),6.01-6.00(d,1H),3.77-3.76(d,4H),3.63-3.60(d,1H),3.54-3.48(t,3H),3.43-3.36(m,2H),3.22(s,3H),3.08-3.07(d,1H),3.00-2.93(t,2H),2.92-2.91(t,1H),2.50-2.46(m,2H),2.02-2.00(t,1H),1.48-1.43(m,2H),0.47-0.44(t,2H),0.11-0.08(m,2H).

Molecular formula C₂₅H₃₂N₆O₄Molecular weight of S512.63 LC-MS (Pos, M/z) ═ 513.2[ M + H⁺].

Example 67 Synthesis of- (1, 1-thiomorpholinyl) -2- ((3S,4S) -4-isobutyl-1- (pyrimidin-5-ylmethyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (Compound 29)

The method comprises the following steps:

step 1 Synthesis of 2- ((3S,4S) -1-benzyl-4-isobutylpyrrol-3-yl) -7- (1, 1-thiomorpholinyl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

reacting 2- (3S,4S) -1-benzyl-4-isobutyl pyrrolidine-3-yl) -7-bromo pyrrolo [2,1-f][1,2,4]Triazin-4 (3H) -one (1.0g,2.33mmol) was dissolved in toluene (20mL), thiomorpholine 1, 1-dioxide hydrochloride (956.2mg,5.592mmol), Pd was added₂(dba)₃(88.0mg,0.093mmol), sodium tert-butoxide (1.35g,13.98mmol) and BINAP (116.3mg,0.186mmol), N₂The reaction was heated to 110 ℃ under reflux overnight with protection. The reaction was monitored by TLC, the reaction mixture was concentrated, water (10mL) was added, dichloromethane was extracted (3 × 25mL), the organic phases were combined, washed with water (2 × 10mL), washed with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was subjected to silica gel column chromatography (DCM: MeOH ═ 200:1-10:1) to give the product (700mg, yield: 62.5%).

Step 2 Synthesis of (3S,4S) -3- (7- (1, 1-thiomorpholino) -4-oxo-3, 4-dihydropyrrolo [2,1-f ] [1,2,4] triazin-2-yl) -4-isobutylpyrrolidine-1-carboxylic acid tert-butyl ester:

dissolving 2- ((3S,4S) -1-benzyl-4-isobutylpyrrol-3-yl) -7- (1, 1-thiomorpholinyl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (380.0mg,0.786mmoL) in dichloromethane (1.5mL) and methanol (1.5mL), adding di-tert-butyl dicarbonate (171.63mg,0.786mmoL) and Pd/C (19.0mg), replacing hydrogen three times, hydrogenating overnight, monitoring the reaction by TLC for completion, filtering with diatomaceous earth, and concentrating the filtrate under reduced pressure to obtain (3S,4S) -3- (7- (1, 1-thiomorpholinyl) -4-oxo-3, 4-dihydropyrrolo [2,1-f ] [1,2,4] triazin-2-yl) -4-isobutylpyrrolidin-1- Tert-butyl formate (620.0mg, crude).

Step 3 Synthesis of 7- (1, 1-thiomorpholinyl) -2- ((3S,4S) -4-isobutylpyrrol-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

tert-butyl (3S,4S) -3- (7- (1, 1-thiomorpholino) -4-oxo-3, 4-dihydropyrrolo [2,1-f ] [1,2,4] triazin-2-yl) -4-isobutylpyrrolidine-1-carboxylate (600mg crude) was dissolved in dichloromethane (5mL), trifluoroacetic acid (2.5mL) was added dropwise at 0 deg.C, the reaction was slowly warmed to room temperature for 2 hours, and the reaction was monitored by TLC for completion. The reaction solution was concentrated under reduced pressure to give 7- (1, 1-thiomorpholinyl) -2- ((3S,4S) -4-isobutylpyrrol-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (620mg crude).

Step 4 Synthesis of 7- (1, 1-thiomorpholinyl sulfide) -2- ((3S,4S) -4-isobutyl-1- (pyrimidin-5-ylmethyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

7- (1, 1-thiomorpholinyl) -2- ((3S,4S) -4-isobutylpyrrol-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (188.76mg) was dissolved in DCM (4mL), pyrimidine-5-carbaldehyde (62.27mg,0.576mmol) and sodium triacetoxyborohydride (254.3mg,1.2mmol) were added, stirring was carried out overnight at room temperature, the reaction was monitored by TLC for completion, a saturated aqueous sodium bicarbonate solution (5mL) was added, extraction was carried out with DCM (3X 10mL), organic phase was combined, water (2X 5mL) was washed, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (DCM: MeOH: 100:1-20:1) to give 7- (1, 1-thiomorpholinyl) -2- ((3S as a white solid, 4S) -4-isobutyl-1- (pyrimidin-5-ylmethyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (130.0mg, yield: 56.03%).

¹HNMR(400MHz,CDCl₃)δ(ppm):9.70(s,1H),9.20(s,1H),8.77(s,2H),6.99-6.98(d,1H),6.02-6.00(d,1H),3.79-3.66(m,6H),3.32-3.28(t,1H),3.07-3.05(d,1H),2.88-2.60(m,1H),2.59-2.56(t,1H),2.42-2.41(d,1H),1.94-1.90(t,1H),1.65-1.60(m,1H),1.49-1.36(m,2H),0.92-0.86(m,6H).

Molecular formula C₂₃H₃₁N₇O₃Molecular weight of S485.61 LC-MS (Neg, M/z) ═ 484.0[ M-H ]]^-.

Example Synthesis of 77- (1, 1-thiomorpholinyl) -2- ((trans) -4-isobutyl-1- ((6- ((methylsulfonyl) methyl) pyridin-3-yl) methyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (Compound 31)

The method comprises the following steps:

step 1, 6- (((methylsulfonyl) oxy) methyl) nicotinic acid methyl ester synthesis:

methyl 6- (hydroxymethyl) nicotinate was dissolved in DCM (10mL), cooled to 0 deg.C, triethylamine (2.43g,12mmol) and methanesulfonyl chloride (2.06g,18mmol) were added sequentially and allowed to slowly warm to room temperature for reaction overnight. TLC monitored the reaction completion, saturated aqueous ammonium chloride (5mL) was added, the layers separated, the aqueous layer extracted with DCM (3X 10mL), the organic layers combined, washed with water (2X 5mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give methyl 6- (((methylsulfonyl) oxy) methyl) nicotinate (3.2g crude).

Step 2, synthesis of 6- ((methylthio) methyl) nicotinic acid:

methyl 6- (((methylsulfonyl) oxy) methyl) nicotinate (2.94g,12mmol) was dissolved in DMF (10mL), and sodium thiomethoxide (1.0g,14.4mmol) was added and reacted at room temperature overnight. TLC to monitor the reaction, vacuum concentrating, adding ethyl acetate and water, separating, extracting the water phase with ethyl acetate, combining the organic phases, washing with water, washing with saturated salt water, drying, filtering, and concentrating to obtain 6- ((methylthio) methyl) nicotinic acid (2.0g crude product).

Step 3, synthesis of methyl 6- ((methylthio) methyl) nicotinate:

6- ((methylthio) methyl) nicotinic acid (2.0g crude) was dissolved in DCM (15mL), cooled to 0 ℃ in an ice bath, oxalyl chloride (4.2g,32.8mmol) was added, DMF (0.05mL) was added at 0 ℃, stirred for 2h at 0 ℃, methanol (20mL) was slowly added dropwise, the reaction was monitored by TLC for completion, concentrated under reduced pressure, and the crude product was subjected to silica gel column chromatography (PE: EA ═ 10:1) to give methyl 6- ((methylthio) nicotinate (140.0mg) as a yellow solid.

Step 4, synthesis of 6- ((methylsulfonyl) methyl nicotinate:

methyl 6- ((methylthio) methyl) nicotinate (140.0mg,0.71mmol) was dissolved in DCM (3mL), cooled to 0 ℃ in an ice bath, m-chloroperoxybenzoic acid (300.0mg,0.852mmol) was added, the temperature was slowly raised to room temperature for reaction for 2 hours, TLC monitored for completion of the reaction, dichloromethane (10mL) and saturated aqueous sodium bicarbonate (10mL) were added, the aqueous phase was extracted with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give methyl 6- ((methylsulfonyl) methyl) nicotinate (180.0mg crude).

Step 5 Synthesis of (6- ((methylsulfonyl) methyl) pyridin-3-yl) methanol:

lithium aluminum hydride (44.9mg,1.18mmol) was added to anhydrous tetrahydrofuran (6mL), cooled to 0 ℃ in an ice bath, a solution of methyl 6- ((methylsulfonyl) methyl) nicotinate (180.0mg) in tetrahydrofuran (3mL) was added, the mixture was stirred at 0 ℃ for 1 hour, TLC monitored for completion of the reaction, water (44.9mg), 10% aqueous NaOH (44.9mg), and water (134.7mg) were added in that order, stirred for 10 minutes, filtered through celite, and the filtrate was concentrated to give (6- ((methylsulfonyl) methyl) pyridin-3-yl) methanol (130.0mg crude).

Step 6, synthesis of 6- ((methylsulfonyl) methyl) nicotinaldehyde:

(6- ((methylsulfonyl) methyl) pyridin-3-yl) methanol (130.0mg crude) was dissolved in dichloromethane (8mL), manganese dioxide (904.2mg,10.4mmol) was added, the mixture was stirred overnight at room temperature, the reaction was monitored by TLC for completion, the mixture was filtered through celite, the filtrate was concentrated, and the crude was isolated by preparative thin layer chromatography to give 6- ((methylsulfonyl) methyl) nicotinaldehyde as a white solid (50.0 mg).

Step 7 Synthesis of 7- (1, 1-thiomorpholinyl) -2- ((trans) -4-isobutyl-1- ((6- ((methylsulfonyl) methyl) pyridin-3-yl) methyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one:

7- (1, 1-Thiomorphinyl) -2- ((trans) -4-isobutylpyrrol-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (41.2mg) was dissolved in DCM (3mL), and 6- ((methylsulfonyl) methyl) nicotinaldehyde (25.0mg,0.126 mmol) and sodium triacetoxyborohydride (55.74mg,0.263mmol) were added and stirred at room temperature overnight. The reaction was monitored by TLC for completion, saturated sodium bicarbonate (5mL) was added, extracted with DCM (3 × 10mL), the organic phases combined, washed with water (2 × 5mL), dried over anhydrous sodium sulfate, filtered, concentrated and the crude product purified by silica gel column chromatography (DCM: MeOH ═ 100:1-20:1) to give 7- (1, 1-thiomorpholinyl) -2- ((trans) -4-isobutyl-1- ((6- ((methylsulfonyl) methyl) pyridin-3-yl) methyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (20.0mg, yield: 33.1%).

¹HNMR(400MHz,DMSO)δ(ppm):11.27(s,1H),8.53(s,1H),7.79-7.77(d,1H),7.48-7.46(d,1H),6.79-6.78(d,1H),6.15-6.14(d,1H),4.61(s,2H),3.66(s,5H),3.01(s,3H),2.96-2.82(m,3H),2.80-2.67(m,2H),2.33-2.29(t,1H),1.53-1.51(m,1H),1.38-1.33(m,2H),0.87-0.83(m,6H).

Molecular formula C₂₆H₃₆N₆O₅S₂576.73 molecular weight, LC-MS (Pos, M/z) ═ 577.3[ M + H⁺].

Example Synthesis of 87- (1, 1-thiomorpholinyl) -2- ((trans) -4-isobutyl-1- (pyridin-4-ylmethyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one (Compound 32)

Step 1: synthesis of 7- (1, 1-thiomorpholinyl) -2- ((trans) -4-isobutyl-1- (pyridin-4-ylmethyl) pyrrolidin-3-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4 (3H) -one

Mixing 7- (1, 1-thiomorpholinyl) -2- ((trans) -4-isobutyl pyrrole-3-yl) pyrrolo [2, 1-f)][1,2,4]Triazin-4 (3H) -one (66mg,0.168mmol,1.0eq) was dissolved in DCM, isonicotinal (21.55mg,0.201mmol,1.2eq) and sodium triacetoxyborohydride (88.86mg,0.419mmol,2.5eq) were added, stirred for 4H, and the reaction was monitored by TLC for completion. Adding NaHCO₃The solution (10mL) and DCM (10mL) were separated, the aqueous phase was extracted with DCM (10mL × 3), the organic phases were combined, dried, concentrated and the crude product was purified by silica gel column chromatography (DCM: MeOH ═ 150:1-5:1) to give the product (46mg, 56.6% yield).

¹HNMR(400MHz,DMSO-d₆)δ(ppm):8.52(m,2H),7.32(m,2H),6.81(m,1H),6.12(m,1H),3.68-3.75(m,5H),2.91-2.95(m,1H),2.81-2.85(m,1H),2.61-2.67(m,2H),2.31-2.36(m,2H),1.25-1.72(m,4H),0.81-0.92(m,6H).

Molecular formula C₂₄H₃₂N₆O₃Molecular weight of S484.62 LC-MS (Neg, M/z) ═ 485.2[ M-H ]⁺].

Biological examples

The invention will be better understood from the following biological examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims. The compounds of the present invention are useful for treating or preventing diseases associated with aberrant expression of PDE 9.

In vitro enzymatic Activity assays for Compounds of the invention

And (3) testing the sample: the compounds of the invention, structures and methods of preparation described in table 1 are as described above.

The experimental method comprises the following steps:

1. in 384 well sample plates, 2. mu.l of 5 XTPDE 9A solution was added per well and 2. mu.l of 1 XTIMAP complete reaction buffer was substituted in the full negative control wells.

2. Mu.l of 2.5 Xthe test drug diluted in step B above was added to each well, and 4. mu.l of 1 XIMAP complete reaction buffer in 2% DMSO was added to the full negative control well and the solvent negative control well.

3. Add 4. mu.l of 2.5 XcGMP substrate working solution per well along the walls of the well.

4. The plates were sealed in the dark, centrifuged at 700rpm × 1min and incubated at room temperature for 60min, with shaking on a shaker for the first 5 min.

5. Mu.l of IMAP binding buffer was added per well along the walls of the wells. And sealing the plate and keeping the plate away from light, centrifuging the whole plate at 700rpm multiplied by 1min, shaking the whole plate for 5min at room temperature in a shaking table, and then incubating the plate for 3h at room temperature.

And 6, reading the fluorescence value of each hole by using a Teacan fluorescence microplate reader.

7. The inhibition rate is the average value of each parallel hole, and the calculation method is as follows: 100- (test drug fluorescence intensity-full negative control well fluorescence intensity) × 100/(solvent negative control well fluorescence intensity-full negative control well fluorescence intensity)Strength). The IC of the test drug was calculated from the data obtained for 10 test drug concentrations using nonlinear sigmoidal regression with GraphPad Prism₅₀The value is obtained.

TABLE 1 in vitro enzymatic inhibitory Activity (IC) of the Compounds of the invention₅₀)

Compound (I)	IC for inhibiting activity of PDE9A50(nM)
		Compound 1	13
Compound 2	52
		Compound 10	16
Compound 15	26
		Compound 16	52
Compound 29	18
		Compound 31	2
Compound 32	5

As can be seen from Table 1, the compounds of the present invention have excellent PDE9A inhibitory activity and are useful for the treatment of diseases associated with abnormal expression of PDE 9.

Claims (5)

1. A compound described by the general formula (III-2), or a pharmaceutically acceptable salt thereof,

wherein the content of the first and second substances,

R₃selected from the group consisting of:

R₄is selected from

Ring a "is selected from the following groups:

2. the compound of claim 1, or a pharmaceutically acceptable salt thereof,

3. a pharmaceutical formulation comprising a compound according to any one of claims 1 to 2, or a pharmaceutically acceptable salt thereof, characterised in that it comprises one or more pharmaceutically acceptable carriers.

4. A pharmaceutical formulation comprising a compound according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof, characterized in that it further comprises one or more second therapeutically active agents which are antimetabolites, growth factor inhibitors, mitotic inhibitors, antitumor hormones, alkylating agents, metals, topoisomerase inhibitors, hormonal drugs, immunomodulators, tumor suppressor genes, cancer vaccines, immune checkpoints or antibodies and small molecule drugs associated with tumor immunotherapy.

5. Use of a compound according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease associated with aberrant expression of PDE9, wherein said disease associated with aberrant expression of PDE9 is:

(a) diseases accessible by inhibition of PDE 9;

(b) CNS disorders;

(c) a disease or condition selected from: sleep disorders, bipolar disorders, metabolic syndrome, obesity, diabetes, hyperglycemia, dyslipidemia, impaired glucose tolerance or a disease of the testis, brain, small intestine, skeletal muscle, heart, lung, thymus or spleen, sickle cell disease.