CN105367576A

CN105367576A - Pyrrolopyrimidine compounds as TLR7 agonists

Info

Publication number: CN105367576A
Application number: CN201410405136.0A
Authority: CN
Inventors: 丁照中; 吴颢; 孙飞; 吴立方; 杨玲
Original assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd
Current assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd
Priority date: 2014-08-15
Filing date: 2014-08-15
Publication date: 2016-03-02

Abstract

The invention relates to pyrrolopyrimidine compounds as TLR7 agonists, and particularly relates to compounds of the formula (I) or pharmaceutically acceptable salts thereof, a preparing method of the compounds, pharmaceutical compositions containing the compounds or the salts, and uses of the compounds or the salts for preparation of antivirus medicines. The formula (I) is shown in the specification.

Description

Pyrrolopyrimidines as TLR7 agonists

Technical Field

The invention relates to a novel pyrrolopyrimidine ring compound serving as a TLR7 agonist or a pharmaceutically acceptable salt thereof, in particular to a compound shown as a formula (I) or a pharmaceutically acceptable salt thereof.

Background

Toll-like receptors are expressed on a variety of immune cells. Toll-like receptors recognize highly conserved structural motifs: a pathogen-associated microbial pattern (PAMP) expressed by a microbial pathogen or a damage-associated molecular pattern (DAMP) released by necrotic cells. Stimulation of Toll-like receptors by corresponding pathogen-associated microbial patterns (PAMPs) or damage-associated molecular patterns (DAMPs) triggers a signaling cascade leading to activation of transcription factors such as AP-1, NF-. kappa.B and interferon regulators (impulse response functions). This results in a variety of cellular responses, including the production of interferons, proinflammatory cytokines and effector cytokines, to generate an immune response. To date, 13 Toll-like receptors have been discovered in mammals. Toll-like receptors 1,2, 4,5 and 6 are expressed primarily on the cell surface, and Toll-like receptors 3, 7, 8 and 9 are expressed in endosomes. Different Toll-like receptors recognize different pathogen-derived ligands. For Toll-like receptor 7(TLR7), it induces secretion of interferon alpha (IFN- α) primarily by plasmacytoid dendritic cell (pDC) expression and ligand recognition. Toll-like receptor 7(TLR7) and Toll-like receptor 8(TLR8) are highly homologous. Thus TLR7 ligand, and in most cases also TLR8 ligand. TLR8 stimulation primarily induces production of cytokines such as tumor necrosis factor alpha (TNF-a) and chemokines. Interferon alpha is one of the main drugs for treating chronic hepatitis b or hepatitis c, and TNF-alpha is a proinflammatory cytokine, and excessive secretion may cause serious side effects. Selectivity to TLR7 and TLR8 is therefore critical for the development of TLR7 agonists for the treatment of viral infectious diseases. Several TLR7 agonists have been reported, such as imiquimod, resiquimod, GS-9620. There remains a great need for new TLR7 agonists with improved selectivity, activity and safety. We have found that a series of novel pyrrolopyrimidine derivatives are agonists of TLR 7. Background research and development references the following journal articles: hoffmann, j.a., Nature,2003,426, p 33-38; akira, S., Takeda, K., and Kaisho, T., annual. Rev. immunology,2003,21, 335-; ulevitch, R.J., Naturereviews: Immunology,2004,4, 512-; coffman, r.l., nat.med.2007,13, 552-559; paul, roethle, j.med, chem.2013,56(18),7324, 7333.

Disclosure of Invention

The invention aims to provide a compound shown as a formula (I) or a pharmaceutically acceptable salt thereof,

formula (I)

Wherein,

L₁、L₂are each independently selected from-O-, -CH₂-、-S-、-NH-、-NHC(＝O)-、-C(＝O)-、-C(＝O)NH-、-S(＝O)-、-S(＝O)₂-、-NHS(＝O)₂-or-S (═ O)₂NH-, wherein said group is optionally substituted with one or more R₄Substitution;

R₁selected from hydrogen, C_1-10Alkyl radical, C_2-10Alkenyl radical, C_2-10Alkynyl, C_3-10Cycloalkyl, 3-10 membered heterocycloalkyl, aryl, heteroaryl, wherein said groups are optionally substituted with one or more R₄Substitution;

R₂selected from hydrogen, halogen, cyano, hydroxy, mercapto, amino, COOH, -CONH₂、C_1-10Alkyl radical, C_2-10Alkenyl radical, C_2-10Alkynyl, C_3-10Cycloalkyl, 3-10 membered heterocycloalkyl, aryl, heteroaryl, wherein said groups are optionally substituted with one or more R₄Substitution;

b is selected from C_3-10Cycloalkyl, 3-10 membered heterocycloalkyl, aryl, heteroaryl;

L₃is selected from C_0-6Alkylene, imino, -O-, -S-, -S (═ O) -or-S (═ O)₂-, wherein said radicals are optionally substituted by one or more R₄Substitution;

R₃selected from hydrogen, amino, C_1-10Alkyl radical, C_2-10Alkenyl radical, C_2-10Alkynyl, C_3-10Cycloalkyl, 3-10 membered heterocycloalkyl, aryl, heteroaryl, wherein said groups are optionally substituted with one or more R₄The substitution is carried out by the following steps,

or R₃、L₃Together with the ortho atoms of the B ring, form a saturated or unsaturated 5-to 8-membered ring, optionally substituted with one or more R₄Substitution;

n is 0, 1,2,3,4 or 5;

R₄selected from halogen, cyano, -R, -OR, -O, -SR, -NR₂NR, -C (halogen)₃-CR (halogen)₂、-CR₂(halogen), -OCN, -SCN, -N ═ C ═ O, -NCS, -NO₂、-NRC(＝O)R、-NRC(＝O)OR、-NRC(＝O)NRR、-C(＝O)NRR、-C(＝O)OR、-OC(＝O)NRR、-OC(＝O)OR、-C(＝O)R、-S(＝O)₂OR、-S(＝O)₂R、-OS(＝O)₂OR、-S(＝O)₂NRR、-S(＝O)R、-NRS(＝O)₂R、-NRS(＝O)₂NRR、-NRS(＝O)₂OR、-OP(＝O)(OR)₂、-P(＝O)(OR)₂-C (═ O) R, -C (═ S) R, -C (═ O) OR, -C (═ S) OR, -C (═ O) SR, -C (═ S) SR, -C (═ O) NRR, -C (═ S) NRR, -C (═ NR) NRR, OR-NRC (═ NR) NRR; r is independently selected from H, C_1-8Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl;

and when L is₁is-CH₂or-NH-, R₃Is not H.

In some embodiments of the compounds of formula (I), L₁、L₂Are each independently selected from-O-, -CH₂-, -S-, -NH-, -C (═ O) -, -S (═ O) -, or-S (═ O)₂-, wherein said radicals are optionally substituted by one or more R₄And (4) substitution. In some embodiments of the compounds of formula (I), L₁、L₂Are each independently selected from-O-, -CH₂-, -S-, -NH-, wherein the radicals are optionally substituted by one or more R₄And (4) substitution. In some embodiments of the compounds of formula (I), L₁、L₂Are each independently selected from-O-, -CH₂-, wherein said radicals are optionally substituted by one or more R₄And (4) substitution.

In some embodiments of the compounds of formula (I), R₁Selected from hydrogen, C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-6Cycloalkyl, 3-6 membered heterocycloalkyl, aryl, heteroaryl, wherein said groups are optionally substituted with one or more R₄And (4) substitution. In some embodiments of the compounds of formula (I), R₁Is selected from C_1-6Alkyl, wherein said group is optionally substituted with one or more R₄And (4) substitution.

In some embodiments of the compounds of formula (I), R₂Selected from hydrogen, halogen, cyano, hydroxy, mercapto, amino, CHO, COOH, -CONH₂、C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-6Cycloalkyl, 3-6 membered heterocycloalkyl, aryl, heteroaryl, wherein said groups are optionally substituted with one or more R₄And (4) substitution. In some embodiments of the compounds of formula (I), R₂Selected from hydrogen, halogen, cyano, hydroxy, amino, -CONH₂、C_1-6Alkyl, wherein said group is optionally substituted with one or more R₄And (4) substitution. In some embodiments of the compounds of formula (I), R₂Selected from hydrogen, cyano, -CONH₂Wherein said group is optionally substituted with one or more R₄And (4) substitution.

In some embodiments of the compounds of formula (I), B is selected from aryl, heteroaryl. In some embodiments of the compounds of formula (I), B is selected from 5-7 membered aryl, 5-7 membered heteroaryl. In some embodiments of the compounds of formula (I), B is selected from phenyl, pyridylpyrimidinyl, pyridazinyl, pyrazinyl, thienyl, thiazolyl, furyl, oxazolyl, thiadiazolyl, isoxazolyl, oxadiazolyl, pyrrolyl, imidazolyl, pyrazolyl, isothiazolyl, triazolyl. In some embodiments of the compounds of formula (I), B is selected from phenyl, pyridinyl.

In some embodiments of the compounds of formula (I), L₃Is selected from C_0-6Alkylene, wherein the radical is optionally substituted by one or more R₄And (4) substitution.

In some embodiments of the compounds of formula (I), R₃Selected from hydrogen, amino, C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-8Cycloalkyl, 3-8 membered heterocycloalkyl, aryl, heteroaryl, wherein said groups are optionally substituted with one or more R₄Substitution; or R₃、L₃Together with the ortho atoms of the B ring, form a saturated or unsaturated 5-to 8-membered ring, optionally substituted with one or more R₄And (4) substitution. In some embodiments of the compounds of formula (I), R₃Selected from hydrogen, amino, C_1-6Alkyl radical, C_3-8Cycloalkyl, 3-8 membered heterocycloalkyl, aryl, heteroaryl, wherein said groups are optionally substituted with one or more R₄Substitution; or R₃、L₃Together with the ortho atoms of the B ring, form a saturated or unsaturated 5-to 8-membered ring, optionally substituted with one or more R₄And (4) substitution.

In some embodiments of the compounds of formula (I), R₄Selected from halogen, cyano, -R, -OR, -O, -SR, -NR₂NR, -C (halogen)₃-CR (halogen)₂、-CR₂(halogen), -OCN, -SCN, -N ═ C ═ O, -NCS, -NO₂、-NRC(＝O)R、-C(＝O)NRR、-C(＝O)OR、-OC(＝O)NRR、-C(＝O)R、-S(＝O)₂OR、-S(＝O)₂R、-OS(＝O)₂OR、-S(＝O)₂NRR、-S(＝O)R、-NRS(＝O)₂R, -C (═ O) OR, OR-C (═ O) NRR. In some embodiments of the compounds of formula (I), R₄Selected from halogen, cyano, -R, -OR, -O, -NR₂NR, -C (halogen)₃-CR (halogen)₂、-CR₂(halogen). In some embodiments of the compounds of formula (I), R₄Selected from halogen, -R, -OR, -O.

In some embodiments, the compound of formula (I) is selected from the following compounds:

or a pharmaceutically acceptable salt thereof.

In another aspect, the invention provides a method of treating a viral infection comprising administering a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof.

In another aspect the invention provides the use of a compound of formula I, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a viral infection.

In some embodiments of the invention, the viral infection is an infection by dengue virus, yellow fever virus, west nile virus, japanese encephalitis virus, tick-borne encephalitis virus, kunjin virus, murray valley encephalitis virus, st. In one embodiment of the invention, the viral infection is a hepatitis viral infection. In one embodiment of the invention, the viral infection is a hepatitis b or c virus infection.

Another aspect of the invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients. The pharmaceutical compositions of the present invention may further comprise one or more additional therapeutic agents.

The pharmaceutical composition of the present invention can be prepared by combining the compound of the present invention or a salt thereof with a suitable pharmaceutically acceptable carrier, and can be formulated, for example, into solid, semi-solid, liquid or gaseous preparations such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, solutions, suppositories, injections, inhalants, gels, microspheres, aerosols, and the like.

Typical routes of administration of the compounds of the present invention or pharmaceutically acceptable salts thereof or stereoisomers thereof or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, transmucosal, enteral, or topical, transdermal, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.

The pharmaceutical compositions of the present invention may be manufactured by methods well known in the art, such as conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, lyophilizing, and the like.

For oral administration, the pharmaceutical compositions may be formulated by mixing the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, slurries, suspensions and the like, for oral administration to a patient.

Solid oral compositions may be prepared by conventional mixing, filling or tableting methods. For example, it can be obtained by the following method: the active compounds are mixed with solid excipients, the resulting mixture is optionally milled, if desired with further suitable auxiliaries, and the mixture is then processed to granules, to give tablets or dragee cores. Suitable excipients include, but are not limited to: binders, diluents, disintegrants, lubricants, glidants, sweeteners or flavoring agents, and the like. Such as microcrystalline cellulose, glucose solutions, gum arabic syrups, gelatin solutions, sucrose and starch pastes; talc, starch, magnesium stearate, calcium stearate or stearic acid; lactose, sucrose, starch, mannitol, sorbitol, or dicalcium phosphate; silicon dioxide; croscarmellose sodium, pregelatinized starch, sodium starch glycolate, alginic acid, corn starch, potato starch, methylcellulose, agar, carboxymethylcellulose, crospovidone, and the like. The dragee cores may optionally be coated, in particular with enteric coatings, according to methods well known in normal pharmaceutical practice.

The pharmaceutical compositions may also be adapted for parenteral administration, as sterile solutions, suspensions or lyophilized products in suitable unit dosage forms. Suitable excipients, such as fillers, buffers or surfactants can be used.

The compounds of formula i or pharmaceutically acceptable salts thereof described herein may be administered by any suitable route and method, for example, orally or parenterally (e.g., intravenously). A therapeutically effective amount of a compound of formula I is from about 0.0001 to 20mg/Kg body weight/day, for example from 0.001 to 10mg/Kg body weight/day.

The frequency of dosage of the compounds of formula I is determined by the individual requirements of the patient, for example 1 or 2 times per day, or more times per day. Administration may be intermittent, for example, wherein a patient receives a daily dose of a compound of formula i over a period of several days, followed by a period of several days or more in which the patient does not receive a daily dose of a compound of formula i.

Related definitions:

the following terms and phrases used herein have the following meanings, unless otherwise indicated. A particular term or phrase, unless specifically defined, should not be considered as indefinite or unclear, but rather construed according to ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commodity or its active ingredient.

The terms "optionally" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not. For example, ethyl is "optionally" substituted with halo, meaning that ethyl may be unsubstituted (CH)₂CH₃) Monosubstituted (e.g. CH)₂CH₂F) Polysubstituted (e.g. CHFCH)₂F、CH₂CHF₂Etc.) or completely substituted (CF)₂CF₃). It will be appreciated by those skilled in the art that any group containing one or more substituents will not incorporate any substitution or substitution pattern which is sterically impossible and/or cannot be synthesized.

C as used herein_m-nMeaning that the moiety has m-n carbon atoms. For example, "C_3-10Cycloalkyl "means that the cycloalkyl group has 3 to 10 carbon atoms. "C_0-6Alkylene "means that the alkylene group has 0 to 6 carbon atoms, and when alkylene has 0 carbon atom, the group is a bond.

Numerical ranges herein refer to each integer in the given range. E.g. "C_1-10By "is meant that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, or 10 carbon atoms.

The term "substituted" means that any one or more hydrogen atoms on a particular atom is replaced with a substituent, so long as the valence of the particular atom is normal and the substituted compound is stable. When the substituent is a keto group (i.e., ═ O), meaning that two hydrogen atoms are substituted, the keto substitution does not occur on the aromatic group.

When any variable (e.g., R) occurs more than one time in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2R, the group may optionally be substituted with up to two R, and there are separate options for R in each case. Furthermore, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

Unless otherwise specified, the term "hetero" means a heteroatom or group of heteroatoms (i.e., a group of atoms containing heteroatoms), i.e., atoms other than carbon and hydrogen, or a group of atoms containing such atoms, the heteroatoms being independently selected from the group consisting of oxygen, nitrogen, sulfur, phosphorus, silicon, germanium, aluminum, boron. In embodiments where two or more heteroatoms are present, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may be different from each other.

The term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

The term "hydroxy" refers to an-OH group.

The term "cyano" refers to the group — CN.

The term "mercapto" refers to the-SH group.

The term "amino" refers to the group-NH₂A group.

The term "alkyl" refers to a straight or branched chain saturated aliphatic hydrocarbon group consisting of carbon and hydrogen atoms, which is attached to the rest of the molecule by a single bond. Non-limiting examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-methylhexyl, -CH₂-cyclopropyl and the like.

The term "alkylene" refers to a saturated straight or branched chain or cyclic hydrocarbon group having 2 identical carbon atoms from the parent alkaneA residue derived from the removal of two hydrogen atoms from a nucleus or two different carbon atoms. Non-limiting examples of alkylene groups include, but are not limited to, methylene (-CH)₂-), 1-ethylidene (-CH (CH)₃) -), 1, 2-ethylene (-CH)₂CH₂-), 1-propylene (-CH (CH)₂CH₃) -), 1, 2-propylene (-CH)₂CH(CH₃) -), 1, 3-propylene (-CH)₂CH₂CH₂-) 1, 4-butylene (-CH₂CH₂CH₂CH₂-) and the like.

The term "imino" refers to-NH-.

The term "alkenyl" refers to a straight or branched chain unsaturated aliphatic hydrocarbon group having at least one double bond, consisting of carbon atoms and hydrogen atoms. Non-limiting examples of alkenyl groups include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, 1, 3-butadienyl, and the like.

The term "alkynyl" refers to a straight or branched chain unsaturated aliphatic hydrocarbon group having at least one triple bond composed of carbon atoms and hydrogen atoms. Non-limiting examples of alkynyl groups include, but are not limited to, ethynyl (-C ≡ CH), 1-propynyl (-C ≡ C-CH)₃) 2-propynyl (-CH)₂-C.ident.CH), 1, 3-butadiynyl (-C.ident.C-C.ident.CH), and the like.

The term "cycloalkyl" refers to a saturated or unsaturated, non-aromatic cyclic hydrocarbon group consisting of carbon and hydrogen atoms, preferably containing 1 or 2 rings. The cycloalkyl group may be a monocyclic, fused polycyclic, bridged or spiro ring structure. Non-limiting examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo [2.2.1] heptyl, spiro [3.3] heptyl, and the like.

The term "heterocycloalkyl" refers to a monocyclic, fused polycyclic, bridged, or spiro ring system of no aromatic character in which some of the ring atoms are selected from N, O, S (O)_n(wherein n is 0, 1 or 2) and the remaining ring atoms are C. Such rings may be saturated or unsaturated (e.g. with one or more double bonds), but not intactA fully conjugated pi-electron system. Examples of 3-membered heterocycloalkyl include, but are not limited to, oxiranyl, thietanyl, cycloazenyl, examples of 4-membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thietanyl, examples of 5-membered heterocycloalkyl include, but are not limited to, tetrahydrofuryl, tetrahydrothienyl, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, 1-dioxoisothiazolidinyl, thiazolidinyl, imidazolidinyl, tetrahydropyrazolyl, pyrrolinyl, dihydrofuranyl, dihydrothienyl, examples of 6-membered heterocycloalkyl include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, 1, 4-thiaoxazolidinyl, 1, 4-dioxanyl, thiomorpholinyl, 1,2-, 1, 4-dithianyl, dihydropyridinyl, tetrahydropyridinyl, morpholinyl, piperazinyl, 1, 4-dithianyl, 1, 4-dioxane, thiomorpholinyl, 1,2-, 1, 4-dithianyl, dihydropyri, Examples of dihydropyranyl, tetrahydropyranyl, thiochromanyl, 7-membered heterocyclic hydrocarbyl include, but are not limited to, azepanyl, oxepanyl, thiepanyl, oxaazabicyclo [2.2.1]Heptyl and azaspiro [3.3]]Heptyl, and the like.

The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated pi-electron system. For example, the aryl group can have 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 6 to 12 carbon atoms. Non-limiting examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, and the like.

The term "heteroaryl" refers to a monocyclic or fused polycyclic ring system containing at least one ring atom selected from N, O, S, the remaining ring atoms being C, and having at least one aromatic ring. Non-limiting examples of heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolinyl, isoquinolinyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothienyl, indolyl, isoindolyl, and the like.

The term "pharmaceutically acceptable" is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

As the pharmaceutically acceptable salt, for example, a metal salt, an ammonium salt, a salt with an organic base, a salt with an inorganic acid, a salt with an organic acid, a salt with a basic or acidic amino acid, and the like can be mentioned. Non-limiting examples of metal salts include, but are not limited to, salts of alkali metals, such as sodium, potassium, and the like; salts of alkaline earth metals such as calcium, magnesium, barium, and the like; aluminum salts, and the like. Non-limiting examples of salts with organic bases include, but are not limited to, salts with trimethylamine, triethylamine, pyridine, picoline, 2, 6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, and the like. Non-limiting examples of salts with inorganic acids include, but are not limited to, salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like. Non-limiting examples of salts with organic acids include, but are not limited to, salts with formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, malic acid, maleic acid, tartaric acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like. Non-limiting examples of salts with basic amino acids include, but are not limited to, salts with arginine, lysine, ornithine, and the like. Non-limiting examples of salts with acidic amino acids include, but are not limited to, salts with aspartic acid, glutamic acid, and the like.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound, which contains an acid or base, by conventional chemical methods. In general, such salts are prepared by the following method: prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid, in water or an organic solvent or a mixture of the two. Generally, nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention. Certain compounds of the present invention may exist in polycrystalline or amorphous form.

Certain compounds of the present invention may have asymmetric carbon atoms (optical centers) or double bonds. Racemates, diastereomers, geometric isomers and individual isomers are all included within the scope of the present invention.

The illustrations of enantiomers, ambiscalemic and scalemic or enantiomerically pure compounds herein are from Maehr, J.chem.Ed.1985,62: 114-120. Unless otherwise indicated, the absolute configuration of a stereocenter is indicated by wedge bonds and dashed bonds. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, they include the E, Z geometric isomer unless otherwise specified. Likewise, all tautomeric forms are included within the scope of the invention.

The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-) -and (+) -enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, as well as racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.

Optically active (R) -and (S) -isomers as well as D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one of the enantiomers of a compound of the invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), diastereomeric salts are formed with an appropriate optically active acid or base, followed by resolution of the diastereomers by fractional crystallization or chromatography, as is well known in the art, and the pure enantiomers are recovered. Furthermore, separation of enantiomers and diastereomers is typically accomplished by using chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamate formation from amines).

The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may be labelled with radioactive isotopes, such as tritium (A), (B), (C³H) Iodine-125 (¹²⁵I) Or C-14(¹⁴C) In that respect All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.

The term "pharmaceutically acceptable carriers" refers to those carriers which do not significantly stimulate the organism and do not impair the biological activity and performance of the active compound. By "pharmaceutically acceptable carrier" is meant an inert substance which facilitates administration of the active ingredient in conjunction with administration of the active ingredient, including, but not limited to, any glidant, sweetener, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersant, disintegrant, suspending agent, stabilizer, isotonicity agent, solvent, or emulsifier acceptable for use in humans or animals (e.g., livestock) as permitted by the national food and drug administration. Non-limiting examples of such carriers include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols, and the like. For additional information on the vector, reference may be made to Remington, the science and practice of pharmacy,21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

The term "excipient" generally refers to a carrier, diluent, and/or vehicle necessary to formulate an effective pharmaceutical composition.

The term "effective amount" or "therapeutically effective amount" with respect to a drug or pharmacologically active agent refers to a sufficient amount of the drug or agent that is non-toxic but achieves the desired effect. For oral dosage forms of the invention, an "effective amount" of one active agent in a composition is the amount required to achieve the desired effect when combined with another active agent in the composition. The determination of an effective amount varies from person to person, depending on the age and general condition of the recipient and also on the particular active substance, and an appropriate effective amount in an individual case can be determined by a person skilled in the art according to routine tests.

The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating a target disorder, disease, or condition.

The compounds of the present invention may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combinations thereof with other chemical synthetic methods, and equivalents thereof known to those skilled in the art, with preferred embodiments including, but not limited to, examples of the present invention.

The chemical reactions of the embodiments of the present invention are carried out in a suitable solvent that is compatible with the chemical changes of the present invention and the reagents and materials required therefor. In order to obtain the compounds of the present invention, it is sometimes necessary for a person skilled in the art to modify or select the synthesis steps or reaction schemes based on the existing embodiments.

An important consideration in any synthetic route planning in the art is the selection of suitable protecting groups for reactive functional groups, such as amino groups in the present invention. GreeneandWuts (protective Groups Inorganic Synthesis, Wileyandsons,1991) is the authority for this aspect for trained practitioners. All references cited herein are incorporated herein in their entirety.

The compounds of general formula (II) according to the invention can be prepared by a person skilled in the art of organic synthesis by scheme 1 using standard methods in the art:

general procedure 1

From 2, 4-dichloro-5H-pyrrolo [3,2-d]Pyrimidine (1-1) (commercial agent), protected by SEM, and then protected by NH₃Substitution to give 2-chloro-5- ((2- (trimethylsilyl) ethoxy) methyl) -5H-pyrrolo [3,2-d]Pyrimidin-4-amine (1-2). Different kinds of alcohols (formula R)₁OH) such as n-butanol, under the action of sodium, to form sodium alkoxide which is then subjected to substitution reaction to form intermediate (1-3). Followed by NBS reaction (R)₅Selected from formaldehyde groups or L optionally carrying a protecting group₃-R₃) To obtain the bromide (1-4). Bromine of bromide (1-4) is exchanged into lithium salt under the action of n-butyllithium, and then the bromide reacts with aldehyde to obtain secondary alcohol (1-5). The secondary alcohol (1-5) is converted through 0-3 steps, and then is reduced by trifluoroacetic acid and triethylsilane, and simultaneously the protecting group is removed, and the final product (II) is generated.

The compounds of general formula (III) according to the invention can be prepared by a person skilled in the art of organic synthesis by scheme 2 using standard methods in the art:

general procedure 2

From intermediate (2-1) (R) prepared according to scheme 1₆Selected from methyl carboxylates) and reacting with NBS to give the bromide (2-2). The bromide (2-2) is further reacted in 1 to 3 steps (e.g. reduction to the aldehyde with DIBAL-H followed by NaBH with pyrrole in methanol solvent₃CN reductive amination) to give another bromide (2-3). Bromide (2-3) in Zn (CN)₂/Zn/Pd₂(dba)₃The 2-cyano compound (2-4) is converted under the conditions of/dppf/DMF. SEM was removed with trifluoroacetic acid to give the final product (III).

It will be appreciated by those skilled in the art that the order of the reaction steps in schemes 1 and 2 may be varied in order to prepare the compounds of the invention and are within the scope of the invention.

For clarity, the invention is further illustrated by examples. The embodiments are not limited to defining or specifying the scope of the invention.

All solvents used in the present invention are commercially available and can be used without further purification. The reaction is generally carried out under inert nitrogen in an anhydrous solvent. Proton nuclear magnetic resonance data were recorded on a Bruker AvanceIII400(400MHz) spectrometer with chemical shifts expressed as (ppm) at tetramethylsilane low field. Mass spectra were measured on an agilent 1200 series plus 6110(& 1956A). LC/MS or ShimadzuMS contain a DAD: SPD-M20A (LC) and Shimadzu Micromass2020 detector. The mass spectrometer was equipped with an electrospray ion source (ESI) operating in either positive or negative mode.

The invention employs the following abbreviations: aq represents aqueous; SEMCl represents (2- (chloromethoxy) ethyl) trimethylsilane; eq represents equivalent; 1,3-DPPP represents 1, 3-bis (diphenylphosphino) propane; DCM represents dichloromethane; PE represents petroleum ether; DMF represents N, N-dimethylformamide; NMP stands for N-methylpyrrolidone; EtOAc for ethyl acetate; i-PrOH represents isopropanol; EtOH stands for ethanol; MeOH is methanol; THF represents tetrahydrofuran; BPO stands for benzoyl peroxide; BOC represents tert-butyloxycarbonyl; HOAc is acetic acid; NaCNBH₃Is sodium cyanoborohydride; LAH is lithium aluminum hydride; 9-BBN is 9-borabicyclononane; MsCl is methanesulfonyl chloride; RT is room temperature; O/N is overnight; boc₂O is di-tert-butyl dicarbonate; TFA is trifluoroacetic acid; TFAA is trifluoroacetic anhydride; TEA is triethylamine; DIBAL-H is diisobutylaluminum hydride; NBS is bromosuccinimide; DPPF is 1,1' -bis (diphenylphosphino) ferrocene; ph₃P is triphenylphosphine; pd (OAc)₂Is palladium acetate; pd (PPh)₃P)₂CL₂Is bis (triphenylphosphine) palladium chloride; pd₂(dba)₃Is tris (benzylidene acetone) dipalladium; XANTPHOS is 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene; n-BuLi is n-butyllithium.

The compound is made by hand orThe software names, and the commercial compounds are under the supplier catalog name.

Using a sample prepared with a ShimadzuSIL-20A autosampler and a Shimadzu Dad: HPLC analysis was performed using an Shimadzu LC20AB system from SPD-M20A detector using an XtimateC18(3M packing, 2.1X300mm) column. Method 0-60AB — 6 min: the elution was started with 100% a (a is 0.0675% TFA in water) and ended with 60% B (B is 0.0625% TFA in MeCN) using a linear gradient, with the entire procedure being 4.2 min followed by 1 min of 60% B. The column was equilibrated to 100:0 for an additional 0.8 minutes for a total run time of 6 minutes. Method for 10-80 AB-6 minutes: elution was started with 90% a (a is 0.0675% TFA in water) and ended with 80% B (B is 0.0625% TFA in acetonitrile) using a linear gradient, with the entire procedure being 4.2 min followed by 80% B for 1 min. The column was equilibrated to 90:10 for an additional 0.8 minutes for a total run time of 6 minutes. The column temperature was 50 ℃ and the flow rate was 0.8 mL/min. The scanning wavelength of the diode array detector is 200-400 nm.

Thin Layer Chromatography (TLC) was performed on a Sanpont-group silica gel GF254, spots were detected by irradiation with a UV light lamp, and in some cases by other methods, in these cases iodine (about 1g iodine was added to 10g silica gel and mixed thoroughly), vanillin (about 1g vanillin dissolved in 100mL 10% H)₂SO₄Prepared in (r)), ninhydrin (available from Aldrich) or special color developer (mixed thoroughly (NH)₄)₆Mo₇O₂₄·4H₂O、5g(NH₄)₂Ce(IV)(NO₃)₆、450mLH₂O and 50mL concentrated H₂SO₄Prepared) thin layer plates were spread and the compounds were examined. Still, w.c. was used; kahn, m.; and Mitra, M.Journarof organic chemistry,1978,43, 2923-. Flash column chromatography or thin layerCommon solvents for chromatography are mixtures of dichloromethane/methanol, ethyl acetate/methanol and hexane/ethyl acetate.

Preparative chromatography was performed on a Gilson-281PrepLC322 system using a Gilson UV/VIS-156 detector using an AgellaVenusalASBPrepC 18,5m, 150X21.2mm column; phenomenex gemini c18,5m, 150x30 mm; boston symmetrixc18,5m, 150x30 mm; or phenomenex synergic18, 4 m, 150x30 mm. Eluting the compound with a low gradient of acetonitrile/water containing 0.05% HCl, 0.25% HCOOH or 0.5% NH at a flow rate of about 25mL/min₃·H₂O, total run time 8-15 minutes.

Detailed Description

The following specific examples are included to provide those skilled in the art with a clear understanding of the invention and are included to provide a further understanding of the invention. They should not be considered as limiting the scope of the invention but merely as being exemplary illustrations and representative of the invention.

Example 1

2-butoxy-7- (3- ((4-methylpiperazin-1-yl) methyl) benzyl) -5H-pyrrolo [3,2-d ] pyrimidin-4-amine

The reaction process comprises the following steps:

example 1 scheme

Step A: 2, 4-dichloro-5H-pyrrolo [3,2-d ] pyrimidine (4 g, 21.4 mmol) was dissolved in anhydrous tetrahydrofuran (30mL) and sodium hydride (1.03 g, 60% mineral oil mixture, 25.6 mmol) was added portionwise to the interior at 0 deg.C. The reaction was stirred at room temperature for 30 minutes, (2- (chloromethoxy) ethyl) trimethylsilane (3.9 g, 23.5 mmol) was added dropwise. After stirring at room temperature for another 2 hours, it was diluted with water (120 ml) and extracted with ethyl acetate (100 ml × 2). The combined organic layers were washed with saturated aqueous sodium carbonate solution and brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent: ethyl acetate/petroleum ether 5% to 10%) to give 2, 4-dichloro-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 h-pyrrolo [3,2-d ] pyrimidine (5.8 g, 85%) as a yellow solid.

MS(ESI)M/Z：318[M+H⁺]。

And B: 2, 4-dichloro-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidine (5 g, 15.8 mmol), isopropanol (15 ml) and ammonia (250 ml) were mixed in a 1000 ml high pressure reactor and stirred at 100 ℃ 110 ℃ for 3 hours. After cooling to room temperature, the mixture was diluted with water (250 ml) and filtered to give 2-chloro-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 h-pyrrolo [3,2-d ] pyrimidin-4-amine (4 g, 85%) without further purification.

MS(ESI)M/Z：299[M+H⁺]。

And C: 2-chloro-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine (4 g, 13.4 mmol) and sodium butoxide (5.15 g, 53.6 mmol) were dissolved in n-butanol (55 ml). The mixture was heated to 100 ℃ under nitrogen and stirred for 8 hours. After cooling to room temperature, the mixture was diluted with water (200 ml) and extracted with ethyl acetate (100 ml × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: ethyl acetate/petroleum ether 15% to 25%) to give 2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine (4.1 g, 91%) as a yellow solid.

MS(ESI)M/Z：337[M+H⁺]。

Step D: 2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine (4 g, 12 mmol) was dissolved in anhydrous tetrahydrofuran (40 mL). NBS (2.2 g, 12.5 mmol) was made up as a saturated solution of anhydrous tetrahydrofuran and added to the solution at less than 0 ℃ over 20 minutes. After the addition was complete, the reaction mixture was stirred at 0 ℃ for 30 minutes, then diluted with brine (150 ml) and extracted with ethyl acetate (100 ml × 3). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether 5% to 15%) to give 7-bromo-2-butoxy-5- (2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine (3.85 g, 78%) as a white solid.

MS(ESI)M/Z：415，417[M+H⁺]。

Step E: to a stirred solution of 7-bromo-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine (3 g, 7.25 mmol) in anhydrous tetrahydrofuran (40ml) at-78 degrees celsius under nitrogen blanket was added n-butyllithium (2.5M, 12 ml, 30 mmol). The reaction mixture was stirred at-78 ℃ for 1 hour. Then, a solution of isophthalaldehyde (1.26 g, 9 mmol) in anhydrous tetrahydrofuran (5 ml) was added slowly. After stirring the mixture at-78 ℃ for a further 30 minutes, it was poured into saturated aqueous ammonium chloride (15 ml) and extracted with ethyl acetate (60 ml. times.2). The combined organic layers were concentrated under reduced pressure and the residue was purified by preparative HPLC to give 1.1 g total of 3- ((4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (hydroxy) methyl) benzaldehyde salt.

MS(ESI)M/Z：471[M+H⁺]。

Step F: to a stirred solution of 3- ((4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (hydroxy) methyl) benzaldehyde (200 mg, 0.43 mmol) and 1-methylpiperazine (87 mg, 0.87 mmol) in ethanol (2.5 ml) was added sodium cyanoborohydride (40 mg, 0.64 mmol) portionwise at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, then diluted with water (10ml) and extracted with ethyl acetate (15 ml × 2). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (3- ((4-methylpiperazin-1-yl) methyl) phenyl) methanol, which was used directly in the next step.

MS(ESI)M/Z：555[M+H⁺]。

Step G: to a stirred solution of (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (3- ((4-methylpiperazin-1-yl) methyl) phenyl) methanol (100 mg) in trifluoroacetic acid (2 ml) was added triethylsilane (0.4 ml) in portions. The reaction mixture was stirred at 55 ℃ under nitrogen for 1 hour and concentrated under reduced pressure. The residue was dissolved in a solution of anhydrous potassium carbonate (100 mg) in methanol (5 ml). The mixture was stirred at 50 ℃ for a further 30 minutes and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative HPLC to give 36 mg of 2-butoxy-7- (3- ((4-methylpiperazin-1-yl) methyl) benzyl) -5 h-pyrrolo [3,2-d ] pyrimidin-4-amine trifluoroacetate.

¹HNMR(Methanol–d4,400MHz):7.33-7.21(m,4H),4.55(t,J＝6.8Hz,2H),4.01(s,2H),3.67(s,2H),3.29-3.24(m,4H),2.87-2.80(m,7H),1.87-1.80(m,2H),1.56-1.49(m,2H),1.02(t,J＝6.8Hz,3H)。

MS(ESI)m/z:409[M+H⁺]。

Example 2

2-butoxy-7- (3- (morpholinomethyl) benzyl) -5H-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (3- (morpholinomethyl) phenyl) methanol was prepared according to example 1, step F, substituting morpholine for 1-methylpiperazine.

LCMS(ESI)m/z:542[M+H⁺]。

And B: 2-butoxy-7- (3- (morpholinomethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine formate was prepared according to the procedure used in example 1, step G.

¹HNMR(Methanol–d4,400MHz):8.41(s,2H),7.35-7.24(m,5H),4.49(t,J＝6.8Hz,2H),4.03(s,2H),3.82(s,2H),3.77-3.75(m,4H),2.77-2.73(m,4H),1.83-1.79(m,2H),1.55-1.49(m,2H),1.01(t,J＝6.8Hz,3H)。

MS(ESI)m/z:396[M+H⁺]。

Example 3

7- (3- (aminomethyl) benzyl) -2-butoxy-5H-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (3- (aminomethyl) phenyl) methanol was prepared according to example 1, step F, ammonium acetate instead of 1-methylpiperazine.

LCMS(ESI)m/z:472[M+H⁺]。

And B: 7- (3- (aminomethyl) benzyl) -2-butoxy-5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure used in example 1, step G.

¹HNMR(Methanol–d4,400MHz):7.31-7.15(m,4H),7.06(s,1H),4.32(t,J＝6.6Hz,2H),4.00(s,2H),3.80(s,2H),1.79-1.73(m,2H),1.56-1.50(m,2H),1.01(t,J＝7.4Hz,3H)。

MS(ESI)m/z:326[M+H⁺]。

Example 4

2-butoxy-7- (3- (pyrrolidin-1-ylmethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (3- (pyrrolidin-1-ylmethyl) phenyl) methanol was prepared as in example 1, step F substituting pyrrolidine for 1-methylpiperazine.

And B: 2-butoxy-7- (3- (pyrrolidin-1-ylmethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine formate was prepared according to the procedure used in example 1, step G.

¹HNMR(Methanol–d4,400MHz):8.50(s,2H),7.41-7.28(m,5H),4.45(t,J＝6.8Hz,2H),4.31(s,2H),4.06(s,2H),3.31-3.29(m,4H),2.10-2.07(m,4H),1.81-1.76(m,2H),1.54-1.49(m,2H),1.01(t,J＝6.8Hz,3H)。

MS(ESI)m/z:380[M+H⁺]。

Example 5

2-butoxy-7- (4- ((3, 3-difluoropyrrolidin-1-yl) methyl) benzyl-5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: 4- ((4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (hydroxy) methyl) benzaldehyde, prepared according to example 1, step E, wherein isophthalaldehyde is replaced with terephthalaldehyde.

LCMS(ESI)m/z:471[M+H⁺]。

And B: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((3, 3-difluoropyrrolidin-1-yl) methyl) phenyl) methanol was prepared according to example 1, step F, substituting 3, 3-difluoropyrrolidine for 1-methylpiperazine.

LCMS(ESI)m/z:562[M+H⁺]。

And C: 2-butoxy-7- (4- ((3, 3-difluoropyrrolidin-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared as in example 1, step G.

¹HNMR(Methanol–d4,400MHz):7.28-7.15(m,4H),7.04(s,1H),4.30(t,J＝6.4Hz,2H),3.97(s,2H),3.59(s,2H),2.88-2.71(m,4H),2.30-2.19(m,2H),1.78-1.71(m,2H),1.55-1.46(m,2H),0.98(t,J＝7.2Hz,3H)。

MS(ESI)m/z:416[M+H⁺]。

Example 6

2-butoxy-7- (4- ((3-fluoropyrrolidin-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((3-fluoropyrrolidin-1-yl) methyl) phenyl) methanol was prepared according to example 5, step B, 3-fluoropyrrolidine instead of 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:544[M+H⁺]。

And B: 2-butoxy-7- (4- ((3-fluoropyrrolidin-1-yl) methyl) benzyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure of example 5, step C.

¹HNMR(Methanol–d4,400MHz):7.30-7.24(m,4H),7.06(s,1H),5.24-5.08(m,1H),4.32(t,J＝6.4Hz,2H),3.99(s,2H),3.69-3.57(m,2H),2.88-2.65(m,4H),2.45-2.43(m,1H),2.25-2.11(m,1H),2.02-1.91(m,1H),1.78-1.73(m,2H),1.57-1.50(m,2H),1.01(t,J＝7.2Hz,3H)。

MS(ESI)m/z:398[M+H⁺]。

Example 7

1- (4- ((4-amino-2-butoxy-5H-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) benzyl) pyrrolidin-3-ol

Step A: 1- (4- ((4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (hydroxy) methyl) benzyl) pyrrolidin-3-ol was prepared according to example 5 step B, replacing 3, 3-difluoropyrrolidine with pyrrolidin-3-ol.

LCMS(ESI)m/z:542[M+H⁺]。

And B: 1- (4- ((4-amino-2-butoxy-5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) benzyl) pyrrolidin-3-ol formate was prepared according to the procedure of example 5, step C.

¹HNMR(Methanol–d4,400MHz):8.43(s,2H),7.45-7.39(m,4H),7.25(s,1H),4.53(m,1H),4.44-4.27(m,2H),4.04(s,2H),3.54-3.47(m,1H),3.38-3.36(m,4H),3.22-3.19(m,1H),2.28-2.24(m,1H),2.05-2.01(m,1H),1.82-1.76(m,2H),1.56-1.50(m,2H),1.01(t,J＝7.2Hz,3H)。

MS(ESI)m/z:396[M+H⁺]。

Example 8

2-butoxy-7- (4- (piperidin-1-ylmethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- (piperidin-1-ylmethyl) phenyl) methanol was prepared according to the procedure for example 5, step B, substituting piperidine for 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:540[M+H⁺]。

And B: 2-butoxy-7- (4- (piperidin-1-ylmethyl) benzyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure of example 5, step C.

¹HNMR(Methanol–d4,400MHz):7.28(d,J＝8.0Hz,2H),7.22(d,J＝8.0Hz,2H),7.04(s,1H),4.30(t,J＝6.6Hz,2H),3.98(s,2H),3.47(s,2H),2.42(s,4H),1.77-1.73(m,2H),1.60-1.57(m,4H),1.52-1.46(m,4H),0.99(t,J＝7.4Hz,3H)。

MS(ESI)m/z:394[M+H⁺]。

Example 9

2-butoxy-7- (4- (morpholinomethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- (morpholinomethyl) phenyl) methanol was prepared according to example 5, step B, morpholine instead of 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:542[M+H⁺]。

And B: 2-butoxy-7- (4- (morpholinomethyl) benzyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure of example 5, step C

¹HNMR(Methanol–d4,400MHz):7.28(d,J＝8.0Hz,2H),7.22(d,J＝8.0Hz,2H),7.03(s,1H),4.29(t,J＝6.6Hz,2H),3.96(s,2H),3.67-3.64(m,4H),3.46(s,2H),2.43(s,4H),1.77-1.72(m,2H),1.55-1.45(m,2H),0.98(t,J＝7.4Hz,3H)。

MS(ESI)m/z:396[M+H⁺]。

Example 10

2-butoxy-7- (4- ((4-methylpiperazin-1-yl) methyl) benzyl) -5 h-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((4-methylpiperazin-1-yl) methyl) phenyl) methanol was prepared as in example 5, step B replacing 3, 3-difluoropyrrolidine with 1-methylpiperazine.

LCMS(ESI)m/z:555[M+H⁺]。

And B: 2-butoxy-7- (4- ((4-methylpiperazin-1-yl) methyl) benzyl) -5 h-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the method in example 5, step C.

¹HNMR(Methanol–d4,400MHz):7.29(d,J＝8.0Hz,2H),7.22(d,J＝8.0Hz,2H),7.04(s,1H),4.31(t,J＝6.6Hz,2H),3.97(s,2H),3.50(s,2H),2.49-2.26(m,11H),1.79-1.72(m,2H),1.56-1.47(m,2H),0.99(t,J＝7.4Hz,3H)。

MS(ESI)m/z:409[M+H⁺]。

Example 11

2-butoxy-7- (4- ((dimethylamino) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((dimethylamino) methyl) phenyl) methanol was prepared according to example 5, step B, dimethylamine instead of 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:500[M+H⁺]。

And B: procedure C procedure for preparation of 2-butoxy-7- (4- ((dimethylamino) methyl) benzyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine formate according to example 5.

¹HNMR(Methanol–d4,400MHz):8.48(s,2H),7.41(s,4H),7.26(s,1H),4.43(t,J＝6.8Hz,2H),4.22(s,2H),4.06(s,2H),2.79(s,6H),1.79(m,J＝6.8Hz,2H),1.55-1.49(m,2H),1.01(t,J＝6.8Hz,3H)。

MS(ESI)m/z:354[M+H⁺]。

Example 12

2-butoxy-7- (4- ((diethylamino) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((diethylamino) methyl) phenyl) methanol was prepared as in example 5, step B, diethylamido 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:528[M+H⁺]。

And B: 2-butoxy-7- (4- ((diethylamino) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylate was prepared according to the procedure in example 5, step C.

¹HNMR(Methanol–d4,400MHz):8.48(s,2H),7.42(s,4H),7.25(s,1H),4.41(t,J＝6.8Hz,2H),4.28(s,2H),4.06(s,2H),3.20-3.15(m,4H),1.82-1.77(m,2H),1.55-1.49(m,2H),1.34(t,J＝6.8Hz,6H),1.01(t,J＝6.8Hz,3H)。

MS(ESI)m/z:382[M+H⁺]。

Example 13

2-butoxy-7- (4- ((dipropylamino) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((dipropylamino) methyl) phenyl) methanol was prepared according to example 5, step B, dipropylamine was prepared instead of 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:556[M+H⁺].

And B: 2-butoxy-7- (4- ((dipropylamino) methyl) benzyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure in example 5, step C.

¹HNMR(Methanol–d4,400MHz):7.29-7.19(m,4H),7.04(s,1H),4.32(t,J＝6.5Hz,1H),3.99(s,2H),3.55(s,2H),2.41-2.37(m,4H),1.78-1.74(m,2H),1.57-1.47(m,6H),1.00(t,J＝7.4Hz,3H),0.87(t,J＝7.4Hz,6H)。

MS(ESI)m/z:410[M+H⁺]。

Example 14

7- (4- (azetidin-1-ylmethyl) benzyl) -2-butoxy-5H-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- (azetidin-1-ylmethyl) phenyl) methanol was prepared according to example 5, step B, azetidine instead of 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:512[M+H⁺]。

And B: 7- (4- (azetidin-1-ylmethyl) benzyl) -2-butoxy-5H-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the method in example 5, step C.

¹HNMR(Methanol–d4,400MHz):7.28(d,J＝8.0Hz,2H),7.18(d,J＝8.0Hz,2H),7.04(s,1H),4.31(t,J＝6.8Hz,2H),3.98(s,2H),3.59(s,2H),3.30-3.27(m,4H),2.15-2.10(m,2H),1.78-1.73(m,2H),1.56-1.52(m,2H),1.01(t,J＝6.8Hz,3H)。

MS(ESI)m/z:366[M+H⁺]。

Example 15

2-butoxy-7- (4- ((3-methoxyazetidin-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((3-methoxyazetidin-1-yl) methyl) phenyl) methanol was prepared according to example 5, step B, substituting 3, 3-difluoropyrrolidine with 3-methoxyazetidine.

LCMS(ESI)m/z:542[M+H⁺]。

And B: 2-butoxy-7- (4- ((3-methoxyazetidin-1-yl) methyl) benzyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure used preparation 5, step C.

¹HNMR(Methanol–d4,400MHz):7.28(d,J＝8.0Hz,2H),7.18(d,J＝8.0Hz,2H),7.04(s,1H),4.31(t,J＝6.8Hz,2H),4.06-4.04(m,1H),3.98(s,2H),3.60(s,2H),3.54-3.52(m,2H),3.24(s,3H),3.04-3.02(m,2H),1.78-1.73(m,2H),1.56-1.52(m,2H),1.01(t,J＝6.8Hz,3H)。

MS(ESI)m/z:396[M+H⁺]。

Example 16

2-butoxy-7- (4- ((4-methyl-1, 4-diazepan-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: ((4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((4-methyl-1, 4-diazepan-1-yl) methyl) phenyl) methanol was prepared as in example 5, step B, substituting 1-methyl-1, 4-diazepan for 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:569[M+H⁺]。

And B: 2-butoxy-7- (4- ((4-methyl-1, 4-diazepan-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine formate was prepared according to the procedure of example 5, step C.

¹HNMR(Methanol–d4,400MHz):8.41(s,3H),7.34-7.24(m,5H),4.52(t,J＝6.8Hz,2H),3.99(s,2H),3.76(s,2H),3.38-3.36(m,2H),3.29-3.27(m,2H),2.95(s,2H),2.87-2.84(m,5H),2.07-2.05(m,2H),1.84-1.80(m,2H),1.55-1.49(m,2H),1.03-0.99(t,J＝8.0Hz,3H)。

MS(ESI)m/z:423[M+H⁺]。

Example 17

2-butoxy-7- (4- ((2, 6-dimethylmorpholinyl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((2, 6-dimethylmorpholinyl) methyl) phenyl) methanol was prepared according to example 5, step B, 2, 6-dimethylmorpholine instead of 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:570[M+H⁺]。

And B: 2-butoxy-7- (4- ((2, 6-dimethylmorpholinyl) methyl) benzyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure of preparation 5, step C.

¹HNMR(Methanol–d4,400MHz):7.30-7.28(d,J＝8.0Hz,2H),7.23-7.21(d,J＝8.0Hz,2H),7.06(s,1H),4.34-4.30(t,J＝8.0Hz,2H),3.99(s,2H),3.69-3.64(m,2H),3.47(s,2H),2.73(d,J＝12.0Hz,2H),1.77-1.70(m,4H),1.54-1.51(m,2H),1.11(d,J＝10.4Hz,6H),1.00(t,J＝8.0Hz,3H).

MS(ESI)m/z:424[M+H⁺]。

Example 18

7- (4- ((1S, 4S) -2-oxa-5-azabicyclo [2.2.1] heptan-5-ylmethyl) benzyl) -2-butoxy-5H-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4- ((1S, 4S) -2-oxa-5-azabicyclo [2.2.1] heptan-5-ylmethyl) phenyl) (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methanol was prepared as in example 5, step B, substituting (1S, 4S) -2-oxa-5-azabicyclo [2.2.1] heptane for 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:554[M+H⁺]。

And B: 7- (4- ((1S, 4S) -2-oxa-5-azabicyclo [2.2.1] heptan-5-ylmethyl) benzyl) -2-butoxy-5 h-pyrrolo [3,2-D ] pyrimidin-4-aminecarboxylic acid salt was prepared according to the procedure in example 5, step C.

¹HNMR(Methanol–d4,400MHz)::8.38(brs,2H),7.45(d,J＝8.4Hz,2H),7.37(d,J＝8.4Hz,2H),7.29(s,1H),4.66(s,1H),4.47(t,J＝6.8Hz,2H),4.36-4.27(m,1H),4.24-4.23(m,2H),4.16-4.13(m,1H),4.04(s,2H),3.82-3.81(m,1H),3.33-3.31(m,2H),2.33-2.29(m,1H),2.14–2.11(m,1H),1.83-1.76(m,2H),1.56-1.48(m,2H),1.01(t,J＝7.2Hz,3H).

MS(ESI)m/z:408[M+H⁺]。

Example 19

2-butoxy-7- (4- ((4-methoxypiperidin-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((4-methoxypiperidin-1-yl) methyl) phenyl) methanol was prepared according to example 4, step B substituting 4-methoxypiperidine for 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:570[M+H⁺]。

And B: 2-butoxy-7- (4- ((4-methoxypiperidin-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylic acid salt was prepared according to the procedure of example 5, step C.

¹HNMR(Methanol–d4,400MHz)::8.45(s,2H),7.43-7.38(m,4H),7.28(s,1H),4.45(t,J＝6.4Hz,2H),4.21(s,2H),4.05(s,2H),3.52-3.53(m,1H),3.33-3.39(m,3H),3.26-3.24(m,2H),3.13-3.10(m,2H),1.99-1.92(m,4H),1.84-1.77(m,2H),1.56-1.50(m,2H),1.01(t,J＝7.2Hz,3H)。

MS(ESI)m/z:424[M+H⁺]。

Example 20

2-butoxy-7- (4- ((4-isopropylpiperazin-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- ((4-isopropylpiperazin-1-yl) methyl) phenyl) methanol was prepared as in example 5, step B replacing 3, 3-difluoropyrrolidine with 1-isopropylpiperazine.

LCMS(ESI)m/z:583[M+H⁺]。

And B: 2-butoxy-7- (4- ((4-isopropylpiperazin-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylate was prepared according to the procedure of example 5, step C.

¹HNMR(Methanol–d4,300MHz)::8.45(s,2H),7.31-7.25(m,5H),4.49(t,J＝8.4Hz,2H),3.99(s,2H),3.64(s,2H),3.42-3.40(m,1H),3.21-3.25(m,4H),2.66-2.82(m,4H),1.84-1.79(m,2H),1.56-1.51(m,2H),1.35(d,J＝8.8Hz,6H),1.04-0.99(t,J＝10.0Hz,3H)。

MS(ESI)m/z:437[M+H⁺]。

Example 21

2-butoxy-7- (4- (pyrrolidin-1-ylmethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (4- (pyrrolidin-1-ylmethyl) phenyl) methanol was prepared according to example 5, step B, pyrrole instead of 3, 3-difluoropyrrolidine.

LCMS(ESI)m/z:526[M+H⁺]。

And B: 2-butoxy-7- (4- (pyrrolidin-1-ylmethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine formate was prepared according to the procedure in example 5, step C.

¹HNMR(Methanol–d4,400MHz):8.41(s,2H),7.46(d,J＝8.0Hz,2H),7.40(d,J＝8.0Hz,2H),7.30(s,1H),4.48(t,J＝6.8Hz,2H),4.33(s,2H),4.05(s,2H),3.32-3.30(m,4H),2.10-2.06(m,4H),1.83-1.89(m,2H),1.55-1.48(m,2H),1.02(t,J＝7.2Hz,3H)。

MS(ESI)m/z:380[M+H⁺]。

Example 22

2-butoxy-7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Route for the preparation of 6- (pyrrolidin-1-ylmethyl) nicotinaldehyde:

step A: to a solution of methyl 6-methylnicotinate (10g, 0.0662 mol) in CCl4(100 ml) was added NBS (13.0 g, 0.0728 mol) and BPO (1.6 g, 0.0066 mol) at room temperature. The reaction mixture was heated to 75 ℃ and stirred for 12 hours. After cooling, water (80 ml) was added and extracted with ethyl acetate (200 ml × 2). The organic layer was successively washed with a saturated aqueous solution of sodium thiosulfate (80 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column on silica gel (eluent: petroleum ether/ethyl acetate-20/1) to give methyl 6- (bromomethyl) nicotinate (5.2 g, 34% yield) as a brown solid.

¹HNMR(CDCl₃,400MHz):9.18(d,J＝1.6Hz,1H),8.32(dd,J₁＝8.0Hz,J₂＝2.0Hz,1H),7.55(d,J＝8.0Hz,1H),4.60(s,2H),3.97(s,3H)。

MS(ESI)m/z:230,232[M+H⁺]。

And B: methyl 6- (bromomethyl) nicotinate (5.0 g, 21.73 mmol) was added portionwise to a solution of pyrrolidine (3.09 g, 43.47 mmol) and triethylamine (3 ml, 21.73 mmol) in dry tetrahydrofuran (100 ml) at 0 ℃. After the addition was complete, the reaction mixture was stirred at room temperature for 16 hours, then diluted with water (80 ml) and extracted with ethyl acetate (100 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column on silica gel (eluent: petroleum ether/ethyl acetate-10/1) to give methyl 6- (pyrrolidin-1-ylmethyl) nicotinate (4.1 g, 86% yield) as a brown solid.

¹HNMR(CDCl₃,400MHz):9.11(d,J＝2.0Hz,1H),8.22(dd,J₁＝8.0Hz,J₂＝2.0Hz,1H),7.48(d,J＝8.0Hz,1H),3.91(s,3H),3.81(s,2H),2.58-2.53(m,4H),1.81-1.77(m,4H)。

MS(ESI)m/z:221[M+H⁺]。

And C: to a stirred solution of methyl 6- (pyrrolidin-1-ylmethyl) nicotinate (3.0 g, 13.62 mmol) in anhydrous tetrahydrofuran (70 ml) was added lithium aluminum hydride (1.03 g, 27.24 mmol) in portions below 0 ℃. The reaction was carried out at about 0 ℃ for 2 hours and at room temperature for a further 30 minutes. TLC showed the reaction disappeared. The mixture was then cooled to 0 ℃ and water (1 ml) was added dropwise very slowly. Then, 15% aqueous sodium hydroxide (1 ml) and extra water (3 ml) were added separately and vigorously stirred. The resulting mixture was filtered. The filtrate was dried over anhydrous Mg2SO4 and concentrated to dryness under reduced pressure to give (6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methanol (2.5 g).

¹HNMR(CDCl₃,400MHz):8.41(d,J＝1.6Hz,1H),7.67(dd,J₁＝8.0Hz,J₂＝2.0Hz,1H),7.37(d,J＝8.0Hz,1H),4.67(s,2H),3.75(s,2H),2.57-2.543(m,4H),1.81-1.76(m,4H)。

Step D: (6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methanol (2.5 g, 13 mmol) was dissolved in anhydrous dichloromethane (50 ml). Manganese dioxide (5.0 g, 58 mmol) was added portionwise at 0 ℃. The reaction mixture was stirred at room temperature for 24 hours and filtered. The filtrate was concentrated in vacuo and the residue was purified by column on silica gel (eluent: 15% ethyl acetate in petroleum ether) to give 6- (pyrrolidin-1-ylmethyl) nicotinaldehyde (2.2 g, crude) as a yellow oil.

LCMS(ESI)m/z:191[M+H⁺]。

Preparation of 2-butoxy-7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine:

example 22 procedure

Step E: (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methanol prepared according to the procedure of example 1, step E, 6- (pyrrolidin-1-ylmethyl) nicotinaldehyde instead of m-benzaldehyde.

LCMS(ESI)m/z:527[M+H⁺]。

Step F: 2-butoxy-7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylate was prepared as a white solid according to the procedure of example 1, step G.

¹HNMR(Methanol–d4,400MHz):8.62(s,1H),8.40(brs,1H),7.77(d,J＝8.0Hz,1H),7.40(d,J＝8.0Hz,1H),7.35(s,1H),4.48(s,2H),4.45(t,J＝6.4Hz,2H),4.08(s,2H),3.42-3.38(m,4H),2.13-2.10(m,4H),1.83-1.76(m,2H),1.55-1.49(m,2H),1.01(t,J＝7.2Hz,3H)。

MS(ESI)m/z:381[M+H⁺]。

Example 23

2-butoxy-7- (3- (2- (pyrrolidin-1-yl) ethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Route to 3- (2- (pyrrolidin-1-yl) ethyl) benzaldehyde:

step A: a solution of methyl 3-bromobenzoate (17.0 g, 79.0 mmol), tributyl (vinyl) stannane (33 g, 102 mmol) and Pd (PPh3)4(4.5 g, 4 mmol) in dioxane (200 mL) was stirred at 110 deg.C for 6 hours under nitrogen blanket and then quenched by addition of 10% aqueous potassium fluoride (100 mL). The resulting mixture was stirred at room temperature for another 10 minutes and extracted with ethyl acetate (150 ml. times.3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent: 25% ethyl acetate in petroleum ether) to give about 15 g of crude methyl 3-vinylbenzoate as a yellow oil.

MS(ESI)m/z:163[M+H⁺]。

And B: to a stirred solution of methyl 3-vinylbenzoate in anhydrous tetrahydrofuran (100 ml) under nitrogen was added 9-BBN (0.5M, 166 ml, 83 mmol) via a dropping funnel and the temperature was kept below-30 ℃. After the addition was complete, the reaction mixture was warmed to room temperature and stirred for 16 hours. Then, the mixture was cooled to-30 ℃ and an aqueous solution of H2O2 (30% by mass, 19 ml) was added dropwise, followed by slowly adding 15% aqueous sodium hydroxide (40ml) dropwise. After stirring the resulting mixture at ambient temperature for a further 1 hour, it was diluted with water (200 ml) and extracted with ethyl acetate (200 ml × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give about 9 g of crude methyl 3- (2-hydroxyethyl) benzoate as a pale yellow oil which was used directly in the next step.

¹HNMR(CDCl₃,400MHz):7.92-7.90(m,2H),7.45-7.37(m,2H),3.92(s,3H),3.89(t,J＝6.5Hz,2H),2.93(t,J＝6.5Hz,2H)。

MS(ESI)m/z:181[M+H⁺]。

And C: to a stirred solution of methyl 3- (2-hydroxyethyl) benzoate (10 g) in dry dichloromethane (90mL) was added methanesulfonyl chloride (34 g, 299 mmol) and triethylamine (12 g, 118 mmol) at about 0 ℃. The reaction was stirred at 0 ℃ for 1 hour, quenched with water (50 ml), and extracted with ethyl acetate (100 ml × 3). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 10% ethyl acetate in petroleum ether) to give 2.7g of methyl 3- (2- ((methylsulfonyl) oxy) ethyl) benzoate as a colorless oil.

MS(ESI)m/z:259[M+H⁺]。

Step D: pyrrolidine (2.3 g, 31.3 mmol) and potassium carbonate (2.2 g, 16 mmol) were dissolved in anhydrous acetonitrile (20 ml) and a solution of methyl 3- (2- ((methylsulfonyl) oxy) ethyl) benzoate (2.7 g, 10.4 mmol) in acetonitrile (5 ml) was added over a period of 10 minutes. The reaction was stirred at 70 ℃ for 16 hours, cooled to room temperature, diluted with water (20 ml) and extracted with ethyl acetate (20 ml × 3). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: methanol/dichloromethane equal to 2% to 5%) to give methyl 3- (2- (pyrrolidin-1-yl) ethyl) benzoate (1.7 g, 71%) as a yellow oil.

MS(ESI)m/z:234[M+H⁺]。

Step E: 3- (2- (pyrrolidin-1-yl) ethyl) benzaldehyde was prepared according to the method of example 22, step C, D.

MS(ESI)m/z:204[M+H⁺]。

Step F: 2-butoxy-7- (3- (2- (pyrrolidin-1-yl) ethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylic acid salt was prepared according to example 22, steps E, F.

¹HNMR(Methanol–d4,400MHz):8.42(s,2H),7.30-7.13(m,5H),4.38(t,J＝6.4Hz,2H),4.01(s,1H),3.41(t,J＝7.6Hz,2H),3.35-3.32(m,4H),3.01(t,J＝7.6Hz,2H),2.09-2.05(m,4H),1.81-1.74(m,2H),1.57-1.48(m,2H),1.01(t,J＝7.6Hz,3H)。

MS(ESI)m/z:394[M+H⁺]。

Example 24

2-butoxy-7- (4- (1- (pyrrolidin-1-yl) ethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Route for preparing 4- (1- (pyrrolidin-1-yl) ethyl) benzaldehyde:

step A: to 4-cyanoacetophenone (4 g, 27.56 mmol) and pyrrolidine (2.94 g, 41.33 mmol) in methanol (100 ml) was added acetic acid (0.5 ml) and sodium cyanoborohydride (5.2 g, 82.67 mmol) with stirring and the temperature was kept below 0 ℃. The reaction was stirred at room temperature for 16 hours, then concentrated under reduced pressure. The resulting oil was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate-1/3) to give 2.8 g of 4- (1- (pyrrolidin-1-yl) ethyl) benzonitrile as a colorless oil.

MS(ESI)m/z:201[M+H⁺]。

And B: to a solution of 4- (1- (pyrrolidin-1-yl) ethyl) benzonitrile (2 g, 10 mmol) in dry toluene (100 ml) at-20 to-10 ℃ was added a solution of DIBAL-H (1M, 20 ml, 20 mmol) and controlled to add over 1 hour. The reaction was stirred for another 3 hours, then quenched with a saturated aqueous solution of ammonium chloride and extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting solid was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate 50/1 to 10/1) to give 4- (1- (pyrrolidin-1-yl) ethyl) benzaldehyde (680 mg, 33.5%) as a colorless oil.

(ESI)m/z:204[M+H⁺]。

And C: 2-butoxy-7- (4- (1- (pyrrolidin-1-yl) ethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylic acid salt was prepared according to the procedure of example 22, Steps E, F.

¹HNMR(Methanol–d4,400MHz):8.50(s,2H),7.44-7.38(m,4H),7.27(s,1H),4.45(t,J＝6.4,2H),4.33-4.28(m,1H),4.04(s,2H),3.37-3.33(m,2H),3.14-3.11(m,2H),2.04-2.02(m,4H),1.83-1.78(m,2H),1.72-1.70(m,3H),1.55-1.49(m,2H),1.01(t,J＝7.4,3H)。

MS(ESI)m/z:394[M+H⁺]。

Example 25

2-butoxy-7- (4- (1-methylpiperidin-4-yl) benzyl) -5 h-pyrrolo [3,2-d ] pyrimidin-4-amine

Preparation route of 4- (4-formylphenyl) piperidine-1-carboxylic acid tert-butyl ester:

step A: a mixture of 4-bromopyridine (3.0 g, 19.0 mmol), (4- (methoxycarbonyl) phenyl) boronic acid (2.63 g, 14.6 mmol), Pd (PPh3)2Cl2(0.35 g, 0.5 mmol) and sodium carbonate (6.91 g, 65.2 mmol) in 1, 2-dimethoxyethane (40ml) was heated to 90 ℃ under a nitrogen atmosphere and stirred for 10 hours. The resulting mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate-6/1 to 2/1) to give methyl 4- (pyridin-4-yl) benzoate (2.7 g, yield: 86.8%) as a white solid.

MS(ESI)m/z:214[M+H⁺]。

And B: to methyl 4- (pyridin-4-yl) benzoate (3.8 g, 17.8 mmol) and PtO₂(0.2 g) in methanol (40ml) 2 ml of hydrochloric acid was added, heated to about 50 ℃ and stirred under hydrogen (50psi) for 16 hours. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure to give crude methyl 4- (piperidin-4-yl) benzoate (4.0 g) as the hydrochloride salt without further purification.

MS(ESI)m/z:220[M+H⁺]。

And C: to a stirred mixed solution of methyl 4- (piperidin-4-yl) benzoate (5.0 g, 22.8 mmol) and potassium carbonate (25.0 g, 182.2 mmol) in tetrahydrofuran (50 ml)/water (50 ml) was added di-tert-butyl dicarbonate (10.0 g, 45.8 mmol) in portions and the temperature was maintained below 10 ℃. After the addition was complete, the reaction mixture was stirred at room temperature for another 0.5 h, then diluted with water (50 ml) and extracted with ethyl acetate (50 ml × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate 6/1 to 1/1) to give tert-butyl 4- (4- (methoxycarbonyl) phenyl) piperidine-1-carboxylate (1.9 g, yield: 26.4%) as a white solid.

¹HNMR(CDCl₃,400MHz):7.98(d,J＝8.4Hz,2H),7.28(d,J＝7.6Hz,2H),4.27(s,1H),3.91(s,3H),2.84-2.68(m,3H),1.85(d,J＝12.8Hz,2H),1.66-1.59(m,2H),1.49(s,9H)。

MS(ESI)m/z:320[M+H⁺]。

Step D: tert-butyl 4- (4-formylphenyl) piperidine-1-carboxylate was prepared according to the procedure for example 22, Steps C, D.

MS(ESI)m/z:312.1[M+Na⁺]。

Step F: 2-butoxy-7- (4- (piperidin-4-yl) benzyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure of example 22, Steps E, F.

MS(ESI)m/z:380.2[M+H⁺]。

Preparation of 2-butoxy-7- (4- (1-methylpiperidin-4-yl) benzyl) -5 h-pyrrolo [3,2-d ] pyrimidin-4-amine:

step G: to a solution of 2-butoxy-7- (4- (piperidin-4-yl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine (100 mg, 0.264 mmol) and HCHO (20 mg, 0.666 mmol) in methanol (5 ml) was added sodium cyanoborohydride (50 mg, 0.796 mmol) after stirring for 5 minutes. The reaction was stirred at room temperature for 0.5 h, diluted with water and extracted with ethyl acetate. The organic layer was concentrated in vacuo and the residue was purified by preparative HPLC to give 7.48 mg of 2-butoxy-7- (4- (1-methylpiperidin-4-yl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine.

¹HNMR(Methanol,400MHz):7.21(d,J＝8.0Hz,2H),7.11(d,J＝8.0Hz,2H),7.00(s,1H),4.32-4.28(m,2H),3.94(s,2H),3.00-2.97(m,2H),2.52–2.47(m,1H),2.32(s,3H),2.19–2.15(m,2H),1.80-1.72(m,6H),1.53-1.48(m,2H),0.98(t,J＝7.4Hz,3H)。

MS(ESI)m/z:394[M+H⁺]。

Example 26

2-butoxy-7- (4- (1-methylpyrrolidin-2-yl) benzyl) -5H-pyrrolo [3,2-d ] pyrimidin-4-amine

Preparation route of tert-butyl 2- (4-formylphenyl) pyrrolidine-1-carboxylate:

step A: to a mixture of NaH (446 mg, 18.6 mmol) in anhydrous tetrahydrofuran (20 ml) at 0 degrees celsius under an atmosphere of N2 was added 1-allyl-pyrrol-2-one (1.14 g, 9.11 mmol). A solution of methyl 4-bromobenzoate in dry tetrahydrofuran (10mL) was then added slowly. The mixture was stirred at 90 ℃ for 2 hours, then cooled to room temperature and diluted with 6N hydrochloric acid. The resulting mixture was stirred at 110 ℃ for 12 hours, then the aqueous phase was washed with ethyl acetate (50 ml). Basified with 1N sodium hydroxide solution to a pH of about 9, and extracted with ethyl acetate (50 ml × 2). The combined organic layers were concentrated to dryness in vacuo to give 2.0 g of 5- (4-bromophenyl) -3, 4-dihydro-2 h-pyrrole as a yellow solid, which was used directly in the next step.

And B: to a stirred solution of 5- (4-bromophenyl) -3, 4-dihydro-2-hydro-pyrrole (2.0 g, 9.0 mmol) in methanol (20 ml) was slowly added sodium borohydride (684 mg, 18.1 mmol) at 0 ℃. After the addition was complete, the reaction mixture was stirred at room temperature for 1 hour. TLC (petroleum ether/ethyl acetate 2:1) showed complete consumption of starting material. The resulting mixture was diluted with water (30 ml). To the mixture in the above step was added potassium carbonate (1.51 g, 10.9 mmol) and Boc2O (2.3 g, 10.5 mmol). After stirring the mixture at 20 ℃ for 2 hours, a thin layer chromatography plate (developing solvent: petroleum ether/ethyl acetate 2/1) showed complete consumption of the starting material. Then, extraction was performed with ethyl acetate (50 ml. times.2), the extract was concentrated under reduced pressure, and the residue was purified by silica gel chromatography to give tert-butyl 2- (4-bromophenyl) pyrrolidine-1-carboxylate (1.5 g, yield: 51.1%) as a yellow solid.

And C: to a stirred solution of tert-butyl 2- (4-bromophenyl) pyrrolidine-1-carboxylate (0.6 g, 1.839 mmol) in anhydrous tetrahydrofuran (20 ml) at-78 deg.c under a nitrogen atmosphere was added n-BuLi (1.5 ml, 2.76 mmol). The reaction mixture was stirred at-78 degrees celsius for 30 minutes, then n.n-dimethylformamide (192 mg, 2.63 mmol) was slowly added to the mixture. The resulting mixture was warmed to room temperature, stirred for an additional 30 minutes and quenched with 3 ml of aqueous sodium bicarbonate. Diluted with water (30ml) and extracted with ethyl acetate (25 ml × 3). The combined organic layers were washed with brine, dried over sodium sulfate, filtered and evaporated to dryness under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether: ethyl acetate ═ 15:1 to 10:1) to give tert-butyl 2- (4-formylphenyl) pyrrolidine-1-carboxylate (0.4g, yield: 79.1%) as colorless oil.

MS(ESI)m/z:276.0[M+1⁺]。

Preparation of 2-butoxy-7- (4- (pyrrolidin-2-yl) benzyl) -5-hydro-pyrrolo [3,2-d ]2 pyrimidin-4-amine:

step D: 2-butoxy-7- (4- (pyrrolidin-2-yl) benzyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure of example 22, steps E, F.

MS(ESI)m/z:366.2[M+1⁺]。

Preparation of 2-butoxy-7- (4- (1-methylpyrrolidin-2-yl) benzyl) -5H-pyrrolo [3,2-d ] pyrimidin-4-amine:

step E: 2-butoxy-7- (4- (1-methylpyrrolidin-2-yl) benzyl) -5H-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure of example 25, step G.

¹HNMR(Methanol-d4,400MHz):7.27(d,J＝8.0Hz,2H),7.22(d,J＝8.0Hz,2H),7.03(s,1H),4.30(t,J＝7.4Hz,2H),3.97(s,2H),3.31-3.19(m,1H),3.07-3.03(m,1H),2.31-2.87(m,1H),2.18-2.15(m,1H),2.13(s,3H),1.89-1.72(m,5H),1.54-1.48(m,2H),0.98(t,J＝7.4Hz,3H)。

MS(ESI)m/z:380[M+1⁺]。

Example 27

1- (4- ((4-amino-2-butoxy-5-hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) phenyl) -4-methylpiperazin-2-one

Preparation of 4- (4-methyl-2-oxopiperazin-1-yl) benzaldehyde:

step A: to a solution of 4-bromo-benzaldehyde (1.8 g, 9.73 mmol), 4-methylpiperazin-2-one (1.44 g, 12.6 mmol), Pd2(dba)3(768 mg, 0.84 mmol), Xantphos (435 mg, 0.75 mmol) and cesium carbonate (5.48 g, 16.8 mmol) in dioxane (30mL) was added water (1 drop). The mixture was stirred at 90 ℃ for 1.5 hours under nitrogen atmosphere. After cooling, the mixture was filtered. The filtrate was concentrated to dryness in vacuo and the residue was purified by silica gel chromatography to give 4- (4-methyl-2-oxopiperazin-1-yl) benzaldehyde (1.8 g, 84.8%) as a white solid.

MS(ESI)m/z:219[M+H⁺]。

Preparation of 1- (4- ((4-amino-2-butoxy-5-hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) phenyl) -4-methylpiperazin-2-one:

and B: 1- (4- ((4-amino-2-butoxy-5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) phenyl) -4-methylpiperazin-2-one was prepared according to the procedure used for example 22, steps E, F.

¹HNMR(Methanol-d4,400MHz)7.36(s,1H),7.30(d,J＝8.4Hz,2H),7.22(d,J＝8.4Hz,2H),4.52(t,J＝6.4Hz,2H),4.02(s,2H),3.72-3.69(m,2H),3.27(s,2H),2.89-2.86(m,2H),2.44(s,3H),1.83-1.79(m,2H),1.54-1.48(m,2H),1.00(t,J＝7.4Hz,3H)。

MS(ESI)m/z:409[M+H⁺]。

Example 28

2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Preparation route of 7-formyl-3, 4-dihydroisoquinoline-2 (1 hydro) -carboxylic acid tert-butyl ester:

step A: trifluoroacetic anhydride (34 g, 0.16 mol) was added dropwise to a solution of 2- (4-bromophenyl) ethylamine (27 g, 0.13 mol) and triethylamine (16.4 g, 0.16 mol) in anhydrous dichloromethane (300 ml) at 0 deg.c under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 1 hour and then diluted with water. The organic layer was separated and concentrated to dryness in vacuo to give N- (4-bromophenylethyl) -trifluoroacetamide (37 g, 96.1%) as a white solid.

MS(ESI)m/z:296,298[M+H⁺]。

And B: to a stirred suspension solution of N- (4-bromophenylethyl) -trifluoroacetamide (37 g, 0.12 mmol) in a mixed acid of concentrated sulfuric acid (200 ml) and acetic acid (300 ml) was added paraformaldehyde (10.2 g, 0.34 mol) in portions. After the addition was completed, the reaction mixture was stirred at room temperature for 12 hours, and then poured into ice water (1 l) and extracted with ethyl acetate (400 ml × 2). The combined organic layers were washed successively with saturated aqueous sodium bicarbonate solution and brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 5% ethyl acetate in petroleum ether) to give 1- (7-bromo-3, 4-dihydroisoquinolin-2 (1 h) -yl) -trifluoroacetone (33 g, 89.3%).

MS(ESI)m/z:308,310[M+H⁺]。

And C: to a solution of 1- (7-bromo-3, 4-dihydroisoquinolin-2 (1 h) -yl) -trifluoroacetone (30 g, 0.1 mol) in anhydrous methylpyrrolidin-2-one (300 ml) was added cuprous cyanide (18 g, 0.2 mol). The reaction mixture was stirred at 180 degrees celsius for 4 hours under a nitrogen atmosphere. After cooling to room temperature, the mixture was slowly poured into ice-water (500 ml) and extracted with ethyl acetate (200 ml × 2). The combined organic layers were washed with water, dried over anhydrous sodium sulfate and concentrated in vacuo to give 25 g of crude 2-trifluoroacetyl-tetrahydroisoquinoline-7-carbonitrile, which was used directly in the next step.

MS(ESI)m/z:255[M+H⁺]。

Step D: 2-three fluoro acetyl four hydrogen isoquinoline-7-nitrile (25 g, 0.1 mol) and potassium carbonate (25 g, 0.18/mol) dissolved in methanol (300 ml) and water (60 ml) mixed solvent, at room temperature for 2 hours. Di-tert-butyldicarbonate (26 g, 0.12 mol) was then added in portions over 10 minutes. After the reaction mixture was stirred at room temperature for another 4 hours, it was diluted with water (200 ml) and extracted with ethyl acetate (200 ml × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (eluent: 5% ethyl acetate in petroleum ether) to give 7-cyano-3, 4-dihydroisoquinoline-2 (1 h) -carboxylic acid tert-butyl ester (14 g, 54%) as a white solid.

MS(ESI)m/z:259[M+H⁺]。

Step E: to a solution of 7-cyano-3, 4-dihydroisoquinoline-2 (1 h) -carboxylic acid tert-butyl ester (1 g, 3.9 mmol) in anhydrous tetrahydrofuran (20 ml) was added dropwise diisobutylaluminum hydride (1M, 6 ml, 6.0 mmol) at-10 ℃ under a nitrogen atmosphere. After the addition was complete, the reaction mixture was stirred at 0 ℃ for 5 hours and quenched with water (0.24 ml). Then, 15% aqueous sodium hydroxide (0.24 ml) was added, and 0.6 ml of water was further added. The resulting mixture was further stirred at room temperature for 15 minutes, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated in vacuo, and the residue was purified by silica gel column chromatography (eluent 10% ethyl acetate petroleum ether solution) to give 7-formyl-3, 4-dihydroisoquinoline-2 (1 h) -carboxylic acid tert-butyl ester (700 mg, 70%) as a yellow oil.

MS(ESI)m/z:262[M+H⁺]。

Preparation of 2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine:

step F: 2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylic acid salt was prepared according to the procedure of example 22, steps E, F.

¹HNMR(Methanol–d4,400MHz):8.49(s,2H),7.23-7.15(m,3H),7.10(s,1H),4.44(t,J＝6.5Hz,2H),4.30(s,2H),3.98(s,2H),3.47(t,J＝6.1Hz,2H),3.08(t,J＝6.1Hz,2H),1.83-1.76(m,2H),1.55-1.49(m,2H),1.01(t,J＝7.4Hz,3H)。

MS(ESI)m/z:352[M+H⁺]。

Example 29

2-butoxy-7- ((2-methyl-1, 2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Preparation of 2-butoxy-7- ((2-methyl-1, 2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine 2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared as described for example 25, step G.

¹HNMR(Methanol–d4,400MHz):7.11-7.09(m,1H),7.03-7.00(m,3H),4.32(t,J＝6.4Hz,2H),3.92(s,2H),3.55(s,2H),2.91-2.88(m,2H),2.73-2.71(m,2H),2.43(s,3H),1.80-1.73(m,2H),1.56-1.52(m,2H),1.01(t,J＝7.6Hz,3H)。

MS(ESI)m/z:366[M+H⁺]。

Example 30

2-butoxy-7- ((2-ethyl-1, 2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Using 2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine as a starting material, 2-butoxy-7- ((2-ethyl-1, 2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amineformate was prepared according to the procedure of example 25, step G.

¹HNMR(Methanol–d4,400MHz):8.43(s,2H),7.25-7.18(m,3H),7.10(s,1H),4.45(t,J＝6.4Hz,2H),4.34(s,2H),3.99(s,2H),3.51(t,J＝6.0Hz,2H),3.32-3.26(m,2H),3.15(t,J＝6.0Hz,2H),1.84-1.77(m,2H),1.58-1.48(m,2H),1.42(t,J＝8.0Hz,3H),1.01(t,J＝6.0Hz,3H)。

MS(ESI)m/z:380[M+H⁺]。

Example 31

2-butoxy-7- ((2-isopropyl-1, 2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Preparation of 2-butoxy-7- ((2-isopropyl-1, 2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine 2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-7-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was used as a starting material and prepared according to the method of example 25, step G.

¹HNMR(Methanol–d4,400MHz):7.10-7.08(m,1H),7.03-7.00(m,3H),4.32(t,J＝6.4Hz,2H),3.93(s,2H),3.70(s,2H),2.90-2.86(m,3H),2.83-2.80(m,2H),1.80-1.73(m,2H),1.56-1.50(m,2H),1.17(d,J＝6.4Hz,6H),1.01(t,J＝7.6Hz,3H)。

MS(ESI)m/z:394[M+H⁺]。

Example 32

2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-6-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

The preparation route of the N-tert-butyloxycarbonyl 1,2,3, 4-tetrahydroisoquinoline-6-formaldehyde comprises the following steps:

step A: to a mixed solution of 6-bromoisoquinoline (10g, 48 mmol) in N, N-dimethylformamide/methanol (V/V ═ 1/1) (200 ml) was added sodium acetate (5.0 g, 61 mmol), triphenylphosphine (3.0 g, 11.4 mmol, and palladium acetate (2.8 g, 12 mmol), and the mixture was charged with 300 kpa of carbon monoxide in an autoclave and heated to 100 degrees celsius, after stirring for 15 hours, completion of the reaction was judged by LC-MS, the reaction product thereof was concentrated by filtering celite (eluting with ethyl acetate), the resulting mixture was concentrated, and the residue was purified by silica gel column chromatography under reduced pressure (eluent: petroleum ether/ethyl acetate ═ 5/1) to give methyl isoquinoline-6-carboxylate (8.9 g, yield: 98%).

MS(ESI)m/z:188[M+H⁺]。

And B: to a stirred solution of methyl isoquinoline-6-carboxylate (10g, 53.5 mmol) in methanol (100 ml) under nitrogen blanket was added acetic acid (2 ml) and PtO₂(200 mg). The mixture was stirred at 40 ℃ for 3 hours under hydrogen atmosphere, and then the catalyst was filtered off through celite and concentrated under vacuum to give methyl 1,2,3, 4-tetrahydroisoquinoline-6-carboxylate (9 g, yield: 88%) without further purification.

MS(ESI)m/z:192[M+H⁺]。

And C: N-tert-Butoxycarbonyl 1,2,3, 4-tetrahydroisoquinoline-6-carboxylic acid methyl ester was prepared according to the method of example 25, step C.

MS(ESI)m/z:292[M+H⁺]。

Step D: N-Boc 1,2,3, 4-tetrahydroisoquinoline-6-carbaldehyde was prepared according to the procedure for example 22, Steps C, D.

MS(ESI)m/z:262[M+H⁺]。

Preparation of 2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-6-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine:

step E: 2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-6-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine was prepared according to the procedure for example 22, steps E, F.

¹HNMR(Methanol–d4,400MHz):7.12-7.09(m,1H),7.08(s,1H),7.04(s,1H),6.96(d,J＝7.6Hz,1H),4.32(t,J＝7.4Hz,2H),3.98(s,2H),3.93(s,2H),3.13(t,J＝6.2Hz,2H),2.85-2.82(m,2H),1.79-1.73(m,2H),1.58-1.48(m,2H),1.01(s,3H)。

MS(ESI)m/z:352[M+H⁺]。

Example 33

2-butoxy-7- ((2-methyl-1, 2,3, 4-tetrahydroisoquinolin-6-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

2-butoxy-7- ((2-methyl-1, 2,3, 4-tetrahydroisoquinolin-6-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine can be prepared by the method of example 25, step G starting from 2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-6-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine.

¹HNMR(Methanol–d4,400MHz):7.10-7.09(m,2H),7.03(s,1H),6.96(d,J＝8.4Hz,1H),4.32(t,J＝6.6Hz,2H),3.93(s,2H),3.60(s,2H),2.92-2.89(m,2H),2.77-2.74(m,2H),2.46(s,3H),1.81-1.73(m,2H),1.58-1.48(m,2H),1.01(t,J＝7.4Hz,3H)。

MS(ESI)m/z:366[M+H⁺]。

Example 34

2-butoxy-7- ((2-ethyl-1, 2,3, 4-tetrahydroisoquinolin-6-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

2-butoxy-7- ((2-ethyl-1, 2,3, 4-tetrahydroisoquinolin-6-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine can be prepared by the method of example 25, step G starting from 2-butoxy-7- ((1,2,3, 4-tetrahydroisoquinolin-6-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine.

¹HNMR(Methanol–d4,400MHz):7.11-7.08(m,2H),7.03(s,1H),6.97(d,J＝8.0Hz,1H),4.32(t,J＝6.6Hz,2H),3.94(s,2H),3.63(s,2H),2.93-2.88(m,2H),2.79-2.76(m,2H),2.65-2.60(m,2H),1.79-1.75(m,2H),1.56-1.52(m,2H),1.21(t,J＝7.2Hz,3H),1.01(t,J＝7.2Hz,3H)。

MS(ESI)m/z:380[M+H⁺]。

Example 35

7-benzyl-2- (2-methoxyethoxy) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

Step A: (4-amino-2- (2-methoxyethoxy) -5- ((2- (trimethylsilylethyl) -5-hydro-pyrrolo [3,2-D ] pyrimidin-7-yl) (phenyl) methanol was prepared as in example 1, steps C, D, E.

MS(ESI)m/z:445[M+H⁺]。

And B: 7-benzyl-2- (2-methoxyethoxy) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylate was prepared as in step G of example 1.

¹HNMR(Methanol–d4,400MHz):8.39(s,1H),7.29-7.19(m,6H),4.61-4.58(m,2H),4.00(s,1H),3.79-3.76(m,2H),3.42(s,3H)。

MS(ESI)m/z:299[M+H⁺]。

Example 36

2- (2-methoxyethoxy) -7- ((6-methylpyridin-3-yl) methyl) -5H-pyrrolo [3,2-d ] pyrimidin-4-amine

2- (2-methoxyethoxy) -7- ((6-methylpyridin-3-yl) methyl) -5H-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylate was prepared as in example 35, Steps A, B.

¹HNMR(Methanol–d4,400MHz):8.34(s,3H),7.66(dd,J＝2.4Hz/J＝8.0Hz,1H),7.31(s,1H),7.24(d,J＝8.0Hz,1H),4.57-4.55(m,2H),4.01(s,2H),3.77-3.75(m,2H),3.41(s,3H),2.51(s,3H)。

MS(ESI)m/z:314[M+H⁺]。

Example 37

7- ((5-Chloropyridin-2-yl) methyl) -2- (2-methoxyethoxy) -5H-pyrrolo [3,2-d ] pyrimidin-4-amine

7- ((5-Chloropyridin-2-yl) methyl) -2- (2-methoxyethoxy) -5H-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylate was prepared as in example 35, Steps A, B.

¹HNMR(Methanol–d4,400MHz):8.45(s,1H),8.40(s,1H),7.77(dd,J＝2.4Hz/J＝8.0Hz,1H),7.38(d,J＝8.0Hz,1H),7.32(s,1H),4.52(t,J＝4.0Hz,2H),4.17(s,2H),3.75(t,J＝4.0Hz,2H),3.42(s,3H)。

MS(ESI)m/z:334[M+H⁺]。

Example 38

2- (2-methoxyethoxy-) -7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

2- (2-methoxyethoxy) -7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine formate was prepared by the method of example 35, steps A, B.

¹HNMR(Methanol–d4,400MHz):8.62(s,1H),8.41(s,2H),7.79-7.76(m,1H),7.36(d,J＝8.4Hz,1H),7.28(s,1H),4.49-4.44(m,4H),4.05(s,2H),3.74-3.72(m,2H),3.39(s,3H),3.33-3.30(m,4H),2.10-2.07(m,4H)。

MS(ESI)m/z:383[M+H⁺]。

Example 39

1- (4- ((4-amino-2- (2-methoxyethoxy) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) phenyl) -4-methylpiperazin-2-one

1- (4- ((4-amino-2- (2-methoxyethoxy) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) phenyl) -4-methylpiperazin-2-one was prepared according to the procedure of example 35, steps a, B.

¹HNMR(Methanol–d4,400MHz):7.35(s,1H),7.31(d,J＝8.4Hz,2H),7.22(d,J＝8.4Hz,2H),4.65-4.62(m,2H),4.01(s,2H),3.77-3.76(m,2H),3.70-3.67(m,2H),3.35(s,3H),3.32-3.28(m,2H),2.90-2.88(m,2H),2.45(s,3H)。

MS(ESI)m/z:411[M+H⁺]。

Example 40

2-butoxy-7- ((5- (pyrrolidin-1-ylmethyl) pyridin-2-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine

2-butoxy-7- ((5- (pyrrolidin-1-ylmethyl) pyridin-2-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-aminecarboxylate was prepared according to the procedure of example 22.

¹HNMR(Methanol–d4,400MHz):8.61(s,1H),8.46(brs,2H),7.91(d,J＝8.0Hz,1H),7.47(d,J＝7.6Hz,1H),7.37(s,1H),4.44(t,J＝6.4Hz,2H),4.35(s,2H),4.22(s,2H),3.33-3.27(m,4H),2.09-2.06(m,4H),1.83-1.76(m,2H),1.57-1.50(m,2H),1.01(t,J＝7.6Hz,3H)。

MS(ESI)m/z:381[M+H⁺]。

EXAMPLE 41

4-amino-2-butoxy-7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidine-6-carbonitrile

Example 41 procedure:

example 41 procedure

Step A: to a solution of 7-bromo-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-4-amine (10.00 g, 24.07 mmol) in anhydrous tetrahydrofuran (200 ml) under a nitrogen atmosphere was added n-BuLi (6.17 g, 96.28 mmol) at-78 ℃. The mixture was stirred at-78 ℃ for 1 hour. A solution of 6-chloronicotinaldehyde (10.22 g, 72.21 mmol) in tetrahydrofuran (200 mL) was then added dropwise. The reaction mixture was stirred at-78 ℃ for another 1 hour, slowly poured into water (150 ml), stirred at room temperature for 20 minutes, and then extracted with ethyl acetate (100 ml × 3). The combined organic phases were washed with saturated brine (50 ml × 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (eluent: petroleum ether/ethyl acetate 5/1 to 1/3) to give (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (6-chloropyridin-3-yl) methanol (5.00 g, 43.45%) as a yellow solid.

¹HNMR(400MHz,CHLOROFORM-d)8.52(d,J＝2.3Hz,1H),7.87(dd,J＝2.4,8.2Hz,1H),7.34(d,J＝8.0Hz,1H),6.65(s,1H),6.14(s,1H),5.97(br.s.,2H),5.39-5.26(m,2H),4.31(t,J＝6.7Hz,2H),3.62-3.49(m,2H),1.86-1.71(m,2H),1.51(qd,J＝7.5,14.9Hz,2H),1.28(t,J＝7.2Hz,1H),1.06-0.87(m,5H),0.00(s,9H)。

MS(ESI)m/z:478[M+H⁺]。

And B: to a solution of (4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) (6-chloropyridin-3-yl) methanol (5.00 g, 10.46 mmol) in trifluoroacetic acid (50 ml) was added triethylsilane (6.08 g, 52.30 mmol) in portions at room temperature. The reaction mixture was stirred at ambient temperature for 24 hours, poured into a saturated aqueous solution of sodium bicarbonate (150 ml) and stirred for a further 20 minutes, then extracted with ethyl acetate (100 ml × 3). The combined organic phases were washed with brine (20 ml × 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (eluent: petroleum ether/ethyl acetate ═ 3/1) to give 2-butoxy-7- ((6-chloropyridin-3-yl) methyl) -5- ((2- (trimethylsilyl) ethoxy) methyl) -5 h-pyrrolo [3,2-d ] pyrimidin-4-amine (2.30 g, 47.59%) as a yellow solid.

¹HNMR(300MHz,CHLOROFORM-d)8.52(d,J＝2.3Hz,1H),7.88(dd,J＝2.4,8.1Hz,1H),7.35(d,J＝8.3Hz,1H),6.64(s,1H),6.14(s,1H),5.89(br.s.,2H),5.40-5.23(m,2H),4.31(t,J＝6.6Hz,2H),3.66-3.47(m,2H),1.88-1.70(m,2H),1.60-1.46(m,2H),1.07-0.82(m,5H),0.00(s,9H)。

MS(ESI)m/z:462[M+H⁺]。

And C: to a solution of 2-butoxy-7- ((6-chloropyridin-3-yl) methyl) -5- ((2- (trimethylsilyl) ethoxy) methyl) -5 h-pyrrolo [3,2-D ] pyrimidin-4-amine (2.30 g, 4.98 mmol) in N, N dimethylformamide (15 ml) was added palladium acetate (111.75 mg, 0.5 mmol), 1, 3-bis-diphenylphosphinopropane (205.30 mg, 0.5 mmol), triethylamine (1.51 g, 14.93 mmol) and methanol (797.43 mg, 24.89 mmol). The suspension was evacuated and charged with carbon monoxide several times. The mixture was heated to 100 degrees celsius and stirred under an atmosphere of carbon monoxide (3 mpa) for 24 hours. Detection on a thin layer chromatography plate (developing solvent: petroleum ether/ethyl acetate: 1/1) indicated complete consumption of the starting material. Insoluble material was filtered and concentrated. The crude product was purified by silica gel chromatography (eluent: with petroleum ether/ethyl acetate ═ 1/1) to give methyl 5- ((4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) picolinate (1.10 g, 45.48%) as a yellow solid.

¹HNMR(400MHz,CHLOROFORM-d)8.76(d,J＝1.8Hz,1H),8.06(d,J＝8.0Hz,1H),7.85(dd,J＝2.0,8.0Hz,1H),6.82(s,1H),5.71(br.s.,2H),5.35(s,2H),4.33(t,J＝6.5Hz,2H),4.19-4.08(m,3H),4.00(s,3H),3.60-3.51(m,2H),1.85-1.74(m,2H),1.53(qd,J＝7.4,15.0Hz,2H),1.28(t,J＝7.2Hz,2H),1.02-0.90(m,5H),0.00(s,9H)。

MS(ESI)m/z:486[M+H⁺]。

Step D: bromosuccinimide (293.18 mg, 1.65 mmol) was added portionwise to a solution of methyl 5- ((4-amino-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) picolinate (800.00 mg, 1.65 mmol) in tetrahydrofuran (10ml) at less than 0 ℃. The reaction mixture was stirred at 0 ℃ for 1 hour, diluted with water (30ml) and extracted with dichloromethane (20 ml × 2). The combined organic phases were dried over magnesium sulfate and concentrated in vacuo. The residue was purified by preparative thin layer chromatography plate to give methyl 5- ((4-amino-6-bromo-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) picolinate (160.00 mg, 17.18%) as a yellow solid.

¹HNMR(400MHz,CHLOROFORM-d)8.83(s,1H),8.03(d,J＝8.0Hz,1H),7.86(d,J＝8.0Hz,1H),5.85(br.s.,2H),5.55(s,2H),4.34(t,J＝6.5Hz,2H),4.10(s,2H),4.00(s,3H),3.71-3.60(m,2H),1.84-1.72(m,4H),1.59-1.47(m,2H),0.98(q,J＝7.8Hz,5H),0.01(s,9H)。

MS(ESI)m/z:565,567[M+H⁺]。

Step E: diisobutylaluminum hydride (56.28 mg, 0.396 mmol) was added dropwise to a stirred solution of methyl 5- ((4-amino-6-bromo-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) picolinate (150.00 mg, 0.266 mmol) in anhydrous tetrahydrofuran (8 ml) at-78 degrees celsius under a nitrogen atmosphere. After the addition was complete, the reaction mixture was stirred at-78 ℃ for 1 hour. Quenched with methanol (5 ml), diluted with water (20 ml) and extracted with ethyl acetate (30ml × 2). The combined organic layers were concentrated to dryness in vacuo to give about 150 mg of crude 5- ((4-amino-6-bromo-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5 hydro-pyrrolo [3,2-d ] pyrimidin-7-yl) methyl) pyridinaldehyde without further purification.

¹HNMR(400MHz,CHLOROFORM-d)10.05(s,1H),8.87(s,1H),7.96-7.80(m,2H),5.72(br.s,2H),5.56(s,2H),4.34(t,J＝6.5Hz,2H),4.12(s,2H),3.71-3.62(m,2H),1.84-1.72(m,2H),1.56-1.48(m,2H),1.06-0.81(m,5H),0.01(s,9H)。

MS(ESI)m/z:535,537[M+H⁺]。

Step F: to a solution of mixture 5- ((4-amino-6-bromo-2-butoxy-5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidin-7 yl) methyl) pyridylaldehyde (150.00 mg, 0.281 mmol), pyrrolidine (29.94 mg, 0.421 mmol), acetic acid (0.2 ml) in tetrahydrofuran (5 ml) was added sodium cyanoborohydride (35.27 mg, 0.561 mmol) and stirred at room temperature for 12 hours. Then, the mixture was poured into an ice/water (1/1, 15 ml) mixture, stirred for 20 minutes, and extracted with ethyl acetate (40ml × 3). The combined organic phases were washed with brine (20 ml × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC to give 150 mg of 6-bromo-2-butoxy-7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5- ((2- (trimethylsilyl) ethoxy) methyl) -5 h-pyrrolo [3,2-d ] pyrimidin-4-amine as a yellow solid.

MS(ESI)m/z:589,591[M+H⁺]。

Step G: 6-bromo-2-butoxy-7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5- ((2- (trimethylsilyl) ethoxy) methyl) -5 h-pyrrolo [3,2-d ] pyrimidin-4-amine (150.00 mg, 254.39 micromole), Pd2(dba)3(23.30 mg, 25.44 micromole), 1,1' -bis (diphenylphosphino) ferrocene (14.10 mg, 25.44 micromole), zinc dicyanide (59.74 mg, 508.78 micromole) and zinc (33.27 mg, 508.78 micromole) were added to anhydrous N, N dimethylformamide (2 mL), displaced with nitrogen, and then heated to 110 ℃ for 3 hours under a nitrogen atmosphere. After cooling, the mixture was diluted with water (30ml) and extracted with ethyl acetate (25 ml × 3). The combined organic phases were washed with brine (30ml), dried over anhydrous sodium sulfate and concentrated in vacuo, and the residue was purified by preparative TLC to give 4-amino-2-butoxy-7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5- ((2- (trimethylsilyl) ethoxy) methyl) -5 h-pyrrolo [3,2-d ] pyrimidine-6-carbonitrile (120 mg, 88.05%)

MS(ESI)m/z：536[M+H⁺]。

Step H: a stirred solution of 4-amino-2-butoxy-7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5- ((2- (trimethylsilyl) ethoxy) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidine-6-carbonitrile (120 mg, 0.224 mmol) in trifluoroacetic acid (5 ml) was stirred at 20 deg.C for 12 h, then concentrated to dryness in vacuo and the residue purified by preparative HPLC to give 8.7 mg of 4-amino-2-butoxy-7- ((6- (pyrrolidin-1-ylmethyl) pyridin-3-yl) methyl) -5-hydro-pyrrolo [3,2-d ] pyrimidine-6-carbonitrile.

¹HNMR(Methanol–d4,400MHz):8.52(s,1H),7.79(d,J＝8.0Hz,1H),7.43(d,J＝8.0Hz,1H),4.33(t,J＝6.8Hz,2H),4.17(s,2H),3.76(s,2H),2.61(s,4H),1.82-1.72(m,6H),1.54-1.49(m,2H),1.02-0.99(t,J＝7.2Hz,3H)。

MS(ESI)m/z:406[M+H⁺]。

Example 42

4-amino-2-butoxy-7- (4- (pyrrolidin-1-ylmethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidine-6-carbonitrile

4-amino-2-butoxy-7- (4- (pyrrolidin-1-ylmethyl) benzyl) -5-hydro-pyrrolo [3,2-D ] pyrimidine-6-carbonitrile is prepared according to the procedure of example 41, with the specific steps shown in example 41, steps A, B, C, D, E, F, G, H.

¹HNMR(Methanol–d4,400MHz):7.34-7.32(d,J＝8.4Hz,2H),7.26-7.24(d,J＝8.4Hz,2H),4.36-4.33(t,J＝6.8Hz,2H),4.13(s,2H),3.62(s,2H),2.57(brs,4H),1.82-1.77(m,6H),1.52-1.49(m,2H),1.00(t,J＝7.2Hz,3H)。

MS(ESI)m/z:405[M+H⁺]。

Example 43

4-amino-2-butoxy-7- (4- (morpholinomethyl) benzyl) -5H-pyrrolo [3,2-d ] pyrimidine-6-carbonitrile

4-amino-2-butoxy-7- (4- (morpholinomethyl) benzyl) -5H-pyrrolo [3,2-D ] pyrimidine-6-carbonitrile hydrochloride was prepared according to the procedure of example 41, with the specific steps shown in example 41, Steps A, B, C, D, E, F, G, H.

¹HNMR(Methanol–d4,400MHz)::7.55(d,J＝7.8Hz,2H),7.43(d,J＝7.8Hz,2H),4.60(t,J＝6.5Hz,2H),4.38(s,2H),4.23(s,2H),4.06-4.02(m,2H),3.80-3.73(m,2H),3.47-3.35(m,2H),3.28-3.14(m,2H),1.89-1.82(m,2H),1.59-1.51(m,2H),1.03(t,J＝7.4Hz,3H)。

LCMS(ESI)m/z:421[M+H⁺]。

Example 44

4-amino-2-butoxy-7- (4- ((4-methylpiperazin-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidine-6-carbonitrile

4-amino-2-butoxy-7- (4- ((4-methylpiperazin-1-yl) methyl) benzyl) -5-hydro-pyrrolo [3,2-D ] pyrimidine-6-carbonitrile hydrochloride was prepared according to the procedure of example 41, with the specific steps shown in example 41, steps A, B, C, D, E, F, G, H.

¹HNMR(Methanol–d4,400MHz)::7.61(d,J＝7.8Hz,2H),7.42(d,J＝7.8Hz,2H),4.60(t,J＝6.5Hz,2H),4.47(s,2H),4.23(s,2H),3.89-3.45(m,8H),3.02(s,3H),1.92-1.80(m,2H),1.61-1.44(m,2H),1.03(t,J＝7.3Hz,3H)。

LCMS(ESI)m/z:434[M+H⁺]。

Example 45

4-amino-2-butoxy-7- (4- (pyrrolidin-1-ylmethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidine-6-carboxamide

Example 45 preparative scheme:

step A: 4-amino-2-butoxy-7- (4- (pyrrolidin-1-ylmethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidine-6-carbonitrile (90 mg, 0.22 mmol) and sodium hydroxide (34 mg, 0.85 mmol) were dissolved in a mixture solvent of methanol (10ml) and water (10ml) and stirred at 80 ℃ for 12 hours. After cooling, it was diluted with water (10ml) and extracted with ethyl acetate (15 ml × 2). The combined organic layers were concentrated to dryness in vacuo and then purified by preparative HPLC to give 10mg of 4-amino-2-butoxy-7- (4- (pyrrolidin-1-ylmethyl) benzyl) -5-hydro-pyrrolo [3,2-d ] pyrimidine-6-carboxamide.

¹HNMR(Methanol–d4,400MHz):7.46(d,J＝8.0Hz,2H),7.32(d,J＝8.0Hz,2H),4.58(t,J＝6.4Hz,2H),4.39(s,2H),4.34(s,2H),3.34-3.32(m,2H),3.18-3.16(m,2H)2.17-2.16(m,2H),2.03-2.00(m,2H),1.86-1.82(m,2H),1.56-1.50(m,2H),1.02(t,J＝7.2Hz,3H)。

MS(ESI)m/z:423[M+H⁺]。

Experimental example 1: toll-like receptor 7 in vitro receptor binding activity screening protocol

Reagent:

HEK-bluehTLR7 cells and HEK-bluehTLR8 cells (from InvivoGen Co.)

DMEM medium

Heat inactivated fetal bovine serum

Anti-mycoplasma reagent Normocin^TM

Bleomycin

Blasticidin

The scheme is as follows:

1.preparation of 96-well Compound plates: compounds were diluted 10 points (2 replicates per spot from column 2 to 11) starting from 10 mmol/l concentration with a 3-fold gradient of DMSO using liquid station POD. Column 12 was filled with 1. mu.l of 5 mg/ml of the positive compound R848 as a positive control, and column 1 with 1. mu.l of DMSO as a negative control. The volume of DMSO contained in each well was 1 μ l.

2. Cells were harvested from the cell culture flasks and the cell density was diluted to 250,000 cells/ml.

3. 200 microliters (50,000 cells/well) of cell suspension was added to the prepared compound plate. The final concentration of DMSO in each well was 0.5%.

4. Placing the culture plate containing the cells and the compound in CO₂Culturing in an incubator for 24 hours under 37 deg.C and 5% CO₂And (4) concentration.

5. After 24 hours of incubation, 20 microliters of supernatant per well of the cell culture plate was removed and transferred to a 96-well clear assay plate. Then add 180. mu.l Quanti-Blue reagent to each well of assay plate and place at 37 ℃ in 5% CO₂Incubate for 1 hour.

After 6.1 hours, the content of alkaline phosphatase in 20. mu.l of the supernatant was measured with a microplate reader OD650 reader.

7. Analysis of the data using Prism software gave the EC for each compound₅₀。

The results of the experiment are shown in table 1:

TABLE 1

Compound (I)	EC₅₀	Compound (I)	EC₅₀	Compound (I)	EC₅₀
						Example 1	C	Example 16	B	Example 31	B
Example 2	C	Example 17	B	Example 32	B
						Example 3	C	Example 18	B	Example 33	B
Example 4	B	Example 19	B	Example 34	B
						Example 5	C	Example 20	B	Example 35	C
Example 6	B	Example 21	B	Example 36	C
						Example 7	B	Example 22	B	Example 37	C
Example 8	B	Example 23	C	Example 38	B
						Example 9	C	Example 24	B	Example 39	B
Example 10	C	Example 25	A	Example 40	B
						Example 11	B	Example 26	B	EXAMPLE 41	A
Example 12	B	Example 27	B	Example 42	A
						Example 13	B	Example 28	B	Example 43	A
Example 14	B	Example 29	B	Example 44	A
						Example 15	B	Example 30	B	Example 45	B

Note: a is more than or equal to 1nM and less than or equal to 100 nM; b is more than 100nM and less than or equal to 1000 nM; c is more than 1000nM and less than or equal to 50 μ M.

Experimental example 2: peripheral blood mononuclear cell assay protocol

The objective of this protocol was to measure cytokine expression levels after 24h stimulation of human Peripheral Blood Mononuclear Cells (PBMC) with existing compounds, without dilution of cell supernatants, and direct measurement of IFN- α and TNF- α levels during the experiment, compounds were first prepared as 20mM DMSO stock solution, diluted 10-fold in cell culture medium, diluted 11-fold in total, 9 dilutions of the compound (maximum compound concentration of 200. mu.M) were added to a 96-well plate at 50. mu.L per well, and fresh human peripheral blood mononuclear cells were inoculated, 150. mu.L per well containing 450,000 cells, and the plates were placed at 37 ℃ and 5% CO₂The plate was centrifuged at 1200rpm for 5 minutes after the incubation was completed for 24 hours in an incubator, and then the supernatant was collected and stored at-20 ℃ to be examined. Detection of cytokines the detection was performed on a flow cytometer using the flow liquid phase multiplex protein quantification technique (CBA) of BD. By using the detection method, the cell factor with the minimum detection limit generated by stimulation is at least 3 times higherThe lowest drug concentration at the sub-level was defined as the mec (minimum effective concentration) value of the compound in the cytokine stimulation experiment.

The results of the experiment are shown in table 2:

TABLE 2

Examples	MEC	Examples	MEC	Examples	MEC
						4	C	28	B	31	B
21	A	29	A	42	A
						22	B	30	B

Note: a is more than or equal to 0.01nM and less than or equal to 1 nM; b is more than 1nM and less than or equal to 10 nM; c is more than 10nM and less than or equal to 100 mu M.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof,

wherein,

L₁、L₂are each independently selected from-O-, -CH₂-、-S-、-NH-、-NHC(＝O)-、-C(＝O)-、-C(＝O)NH-、-S(＝O)-、-S(＝O)₂-、-NHS(＝O)₂-or-S (═ O)₂NH-whereinSaid group being optionally substituted by one or more R₄Substitution;

n is 0, 1,2,3,4 or 5;

and when L is₁is-CH₂or-NH-, R₃Is not H.

2. The compound of claim 1, wherein L is₁、L₂Are each independently selected from-O-, -CH₂-, -S-, -NH-, -C (═ O) -, -S (═ O) -, or-S (═ O)₂-, wherein said radicals are optionally substituted by one or more R₄And (4) substitution.

3. The compound of claim 2, wherein L is₁、L₂Are each independently selected from-O-, -CH₂-, wherein said radicals are optionally substituted by one or more R₄And (4) substitution.

4. The compound of claim 1, wherein R is₁Selected from hydrogen, C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-6Cycloalkyl, 3-6 membered heterocycloalkyl, aryl, heteroaryl, wherein said groups are optionally substituted with one or more R₄And (4) substitution.

5. The compound of claim 4, wherein R is₁Is selected from C_1-6Alkyl, wherein said group is optionally substituted with one or more R₄And (4) substitution.

6. The compound of claim 1, wherein R is₂Selected from hydrogen, halogen, cyano, hydroxy, amino, -CONH₂、C_1-6Alkyl, wherein said group is optionally substitutedOne or more R₄And (4) substitution.

7. The compound of claim 6, wherein R is₂Selected from hydrogen, cyano, -CONH₂Wherein said group is optionally substituted with one or more R₄And (4) substitution.

8. The compound of claim 1, wherein B is selected from the group consisting of aryl and heteroaryl.

9. The compound of claim 8, wherein B is selected from phenyl and pyridyl.

10. The compound of claim 1, wherein L is₃Is selected from C_0-6Alkylene, wherein the radical is optionally substituted by one or more R₄And (4) substitution.

11. The compound of claim 1, wherein R is₃Selected from hydrogen, amino, C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-8Cycloalkyl, 3-8 membered heterocycloalkyl, aryl, heteroaryl, wherein said groups are optionally substituted with one or more R₄Substitution; or R₃、L₃Together with the ortho atoms of the B ring, form a saturated or unsaturated 5-to 8-membered ring, optionally substituted with one or more R₄And (4) substitution.

12. The compound of claim 1, wherein R is₄Selected from halogen, cyano, -R, -OR, -O, -SR, -NR₂NR, -C (halogen)₃-CR (halogen)₂、-CR₂(halogen), -OCN, -SCN, -N ═ C ═ O, -NCS, -NO₂、-NRC(＝O)R、-C(＝O)NRR、-C(＝O)OR、-OC(＝O)NRR、-C(＝O)R、-S(＝O)₂OR、-S(＝O)₂R、-OS(＝O)₂OR、-S(＝O)₂NRR、-S(＝O)R、-NRS(＝O)₂R, -C (═ O) OR, OR-C (═ O) NRR.

13. The compound of claim 12, wherein R is₄Selected from halogen, cyano, -R, -OR, -O, -NR₂NR, -C (halogen)₃-CR (halogen)₂、-CR₂(halogen).

14. A compound represented by the formula:

or a pharmaceutically acceptable salt thereof.

15. Use of a compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a viral infection.

16. A pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients.