pyrazol heteroaryl derivatives, methods of preparation and medical use thereof

专利摘要:
A pyrazol heteroaryl derivative, a method of preparation and medical use thereof is disclosed. in particular, the present invention relates to a novel pyrazol-heteroaryl derivative as shown in general formula (i), a method of preparation thereof and a pharmaceutical composition containing the derivative and its use as a therapeutic agent, in particular as a t7r agonist, wherein each substituent in the general formula (i) is defined in the description.
公开号:BR112019010182A2
申请号:R112019010182
申请日:2017-11-27
公开日:2019-09-17
发明作者:Shu Chunfeng；He Feng；Zhang Guobao；Hu Qiyue；Tao Weikang
申请人:Jiangsu Hengrui Medicine Co；Shanghai hengrui pharmaceutical co ltd；
IPC主号:

专利说明:

Descriptive Report of the Invention Patent for PIRAZOL-HETEROARY DERIVATIVES, METHODS OF PREPARATION AND MEDICAL USE OF THE SAME
FIELD OF INVENTION [001] The present invention relates to a new pyrazoleheteroaryl derivative of formula (I), a method for preparing the same and a pharmaceutical composition comprising them, as well as the use of them as a therapeutic agent, particularly as a TLR7 agonist.
BACKGROUND OF THE INVENTION [002] Toll-like receptors (TLRs) are a class of important protein molecules involved in innate immunity. TLRs are isolated, non-catalytic, membrane receptors, normally expressed in sentinel cells, such as macrophages and dendritic cells and can recognize structurally conserved molecules produced by microbes. Once these microbes have crossed physical barriers, such as the intestinal or skin mucosa, they are recognized by TLRs, which activate immune cell responses (Mahla, RS. Et al., Front Immunol. 4: 248 (2013 )). The immune system's ability to widely recognize pathogenic microorganisms is partly due to the widespread presence of toll-like immunoreceptors (TLRs).
[003] There are at least ten different TLRs in mammals. Ligands and corresponding signaling cascades were
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2/97 identified for some of these receivers. The TLR7 is a member of the subgroup of TLRs (TLRs 3, 7, 8 and 9), located in the endosornal compartment of cells that are specialized for detecting non-proper nucleic acids. TLR7 plays a key role in antiviral defense through ssRNA recognition (Diebold S. S. et al, Science, 2004: 303, 1529-1531; and Lund J. M. et al, PNAS, 2004: 101, 5598-5603). TLR7 has a restricted expression profile in humans and is expressed predominantly by B cells and plasmacytoid dendritic cells (pDC) and, to a lesser extent, by monocytes. Plasmacytoid DCs are a unique population of lymphoid-derived dendritic cells (0.2-0.8% peripheral blood mononuclear cells (PBMCs)), which are cells producing high levels of interferon alpha (IFNa) ) and interferon beta (ΙΓΝβ) in response to viral infections (Liu YJ, Annu. Rev. Immunol., 2005: 23, 275-306).
[004] Various diseases and disorders are related to abnormalities in TLRs, such as melanoma, non-small cell lung carcinoma, hepatocellular carcinoma, basal cell carcinoma, renal cell carcinoma, myeloma, allergic rhinitis, asthma, chronic obstructive pulmonary disease (COPD), ulcerative colitis, liver fibrosis and viral infections such as HBV, virus Flaviviridae, HCV, HPV, RSV, SARS, HIV or influenza. Therefore, the use of a TLR agonist to treat related diseases is very promising.
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3/97 [005] Since TLR7 and TLR8 are highly homologous, the TLR7 ligand in most cases is also that of TLR8. TLR8 stimulation mainly induces cytokine production, such as tumor necrosis factor α (TNF-α) and chemokine. Α interferon is one of the main drugs for the treatment of chronic hepatitis B or hepatitis C, while TNF-α is a pro-inflammatory cytokine, and its excessive secretion can cause serious side effects. Therefore, selectivity for TLR7 and TLR8 is critical to the development of the TLR7 agonist in the treatment of viral infectious diseases.
[006] Currently there are patent applications related to TLR7 agonists, such as W02005025583, W02007093901, WO2008011406, WO2009091032, WO2010077613, WO2010133882, W02011031965 and WC2012080730. However, there is still a need to continue to develop TLR7 agonists that are safer and more therapeutically effective.
[007] In view of the above technical problems, the present invention provides a pharmaceutical compound having a concentration of lesser onset, better selectivity (selective for TLR7 and no activation effect on TLR8), more effective activation effect and at the same time, due while a weak inhibitory effect on CYP is weak, it is a safer and more effective TLR7 agonist.
Summary of the invention [008] The object of the present invention is to provide a compound of formula (I):
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or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, where:
ring A is selected from the group consisting of cycloalkyl, heterocyclyl, aryl and heteroaryl;
G is CH or N;
X ¹ is an alkylene or S (O) m group, where the alkylene is optionally substituted by one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl and heterocyclyl;
L ¹ is selected from the group consisting of -NR ⁴ -, -0-, -S-, -C (0) -, -C (O) -OR ⁴ , -S (0) _m -, -N (R ⁴ ) C (O) -, -C (O) N (R ⁴ ) -,
-N (R ⁴ ) S (0) 2-, -S (O) 2N (R ⁴ ) - and a covalent bond;
R ¹ is selected from the group consisting of alkyl, alkoxy, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, where alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally replaced by
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5/97 one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ^S R ⁷ and -C (O) NR ⁶ R ⁷ ;
each R ² is the same or different and each is independently selected from the group consisting of hydrogen, halogen, alkyl, alkoxy, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl, where alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl , aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ^S R ⁷ and -C (O) NR ⁶ R ⁷ ;
L ² is alkylene or a covalent bond, where the alkylene is optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl , aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ^S R ⁷ and -C (O) NR ⁶ R ⁷ ;
R ³ is selected from the group consisting of haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ e-0 (0) NR ⁶ R ⁷ , in which cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently
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6/97 optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁸ , -C (O) R ⁸ , -S (O) _m R ⁸ , -NR ⁹ R ¹⁰ and -C (O) NR ⁹ R ¹⁰ ;
R ⁴ is selected from the group consisting of hydrogen, alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;
R ⁵ is selected from the group consisting of hydrogen, alkyl, haloalkyl, amino, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl;
R ⁶ and R ⁷ are identical or different if each is independently selected from the group consisting of hydrogen, alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C (O) R ⁸ , -S (O) _m R ⁸ and -C (O) NR ⁹ R ¹⁰ , wherein the alkyl, cycloalkyl, heterocyclyl, arylheteroaryl are each independently optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, amino, cyano , nitro, hydroxy hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;
or, R ⁶ and R ⁷ together with the nitrogen to which they are attached, form a heterocyclyl, where the heterocyclyl optionally contains one or two identical or different heteroatoms selected from the group consisting of N, 0 and S, in addition to a nitrogen atom, and the heterocyclyl is optionally
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7/97 substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;
R ⁸ is selected from the group consisting of hydrogen, alkyl, haloalkyl, amino, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl;
R ⁹ and R ¹⁰ are identical or different and are each independently selected from the group consisting of hydrogen, alkyl, haloalkyl, amino, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl;
neO, 1, 2, 3 or 4; and m is 0, 1 or 2.
[009] In a preferred embodiment of the present invention, in the compound of formula (I), R ³ is heterocyclyl, and the heterocyclyl is optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, amino , cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl.
[010] In a preferred embodiment of the present invention, in the compound of formula (I), R ³ is -NR ⁶ R ⁷ , and R ⁶ and R ⁷ together with the nitrogen to which they are attached, form a heterocyclyl, in that the heterocyclyl optionally contains one or two identical or different heteroatoms selected from the group consisting of N, O and S in addition to a nitrogen atom
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8/97 and the heterocyclyl is optionally substituted with one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl.
[011] In a preferred embodiment of the present invention, in the compound of formula (I), ring A is selected from the group consisting of phenyl and pyridyl.
[012] In a preferred embodiment of the present invention, in the compound of formula (I), pyridyl is selected from the group consisting of [013] In a preferred embodiment of the present invention, in the compound of formula (I), X ~ is an alkylene group.
[014] In a preferred embodiment of the present invention, the compound of formula (I) is a compound of formula (II):
nh ₂
or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a salt
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9 / 9Ί pharmaceutically acceptable thereof, wherein G, L ¹ - L ² , R ¹ ~ R ² , R ⁶ ~ R ^{7 and} n are as defined in the formula [015] In a preferred embodiment of the present invention, the compound of formula (I), where G is N.
[016] In a preferred embodiment of the present invention, in the compound of formula (I), L ² is alkylene.
[017] In a preferred embodiment of the present invention, the compound of formula (I) is a compound of formula (III):
NH
or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, where:
s is 0, 1 or 2;
L ¹ , R ⁴ ~ R ^{2 and} n are as defined in formula (I).
[018] In a preferred embodiment of the present invention, in the compound of formula (I), L ¹ is selected from the group consisting of -O-, -NR ⁴ -, -C (O) - and -C (O ) N (R ⁴ ) -, and R ⁴ is hydrogen or alkyl.
[019] In a preferred embodiment of this
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10/97 invention, in the compound of formula (I), R ¹ is alkyl optionally substituted by one or more alkoxy groups.
[020] In a preferred embodiment of the present invention, in the compound of formula (I), each R ² is the same or different and each is, independently, hydrogen or halogen.
[021] Typical compounds of the present invention include, but are not limited to:
Example No. Structure and name of the compound 1 nh ₂ 1 N 1 6-Butoxy-l- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH -pyrazolo [3,4-d] pyrimidin-4-amine 2 nh ₂ ν'Χλ, _ _ II J, ^{N NN} vz = Π 2 1- (4- (Azetidin-1-ylmethyl) benzyl) -6-butoxy-1H-pyraz olo [3,4-d] pyrimidin-4-amine 3 nh ₂ N _ JI X 'NN ___ < 3 6-Butoxy-l- (4- (piperidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine
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4 nh ₂ _ _ II JN —N / 4 W 6-Butoxy-l- (3- (pyrrolidin-1-ylmethyl) benzyl) -IH -pyrazolo [3,4-d] pyrimidin-4-amine 5 nh ₂ _ Ji JL ' ^N 5 -' 1- (3- (Azetidin-1-ylmethyl) benzyl) -6-butoxy-1H-pi razolo [3,4-d] pyrimidin-4-amine 6 nh ₂ ^N Ί ^ ν. Cf O 6-Butoxy-l- (3- (piperidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 7 nh ₂ Ο Λ JL ' ^N / X 6- (2-Methoxyethoxy) -1- (4- (pyrrolidin-1-ylmethyl) benzi1) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 8 nh _{2 n} 6 - ((1-Methoxypropan-2-yl) oxy) -1- (4- (pyrrolidin-1-yl methyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine
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9 nh ₂ 11 a, ^N 6-Butoxy-l- (3-fluoro-4- (pyrrolidin-l-ylmethyl) benzi1) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 10 nh ₂ _ _ Ji X - ^N NN __ f 10 hb-Butyl-l- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidine-4,6-diamine 11 nh ₂ Η Ϊ Vn O Xfu 11 4-Amino-N-propyl-l- (4- (pyrrolidin-l-ylmethyl) benzi1) -IH-pyrazolo [3,4-d] pyrimidine-6-carboxamide 12 nh ₂ _ Ji JL - ^N 12 1- (4-Amino-1- (4-pyrrolidin-1-ylmethyl) benzyl) -IH-pi razolo [3,4-d] pyrimidin-6-yl) pentan-1-one
or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or the pharmaceutically acceptable one of [022] In another aspect, to a compound of formula (I-C):
mixture of them, or even a salt.
present invention relates to
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or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, where:
W is an amino protecting group, preferably tert-butoxycarbonyl, acetyl, benzyl, alii or p-methoxybenzyl;
X is halogen, preferably chlorine;
the ring A, G, X ¹ , L ² , R ² ~ R ^{3 and} n are as defined in the formula (D.
[023] The compounds of formula (I-C) include, but are not limited to:
No. ofExample Structure and name of the compound le XXnHN1 n 6-Chloro-N- (4-methoxybenzyl) -1- (4- (pyrrolidin-1-imethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine
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2f Ϊ JN CI ^ N ^N __ | —I W 2f 1- (4- (Azetidin-1-ylmethyl) benzyl) -6-chloro-7V- (4-m ethoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amin at 2f 3c pp _NH II JN CI ^ NN _{_} ___ < 3c 6-Chloro-7V- (4-methoxybenzyl) -1- (4- (piperidin-1-yl methyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amiat 3c 4b ^N TA, ii 1 n CI ^ N ^ N 4b 6-Chloro-7V- (3-methoxybenzyl) -1- (4- (pyrrolidin-1-imethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 4b 5e / VAh II JN. CI ^^ NN __ / N ₅ e 1- (3- (Azetidin-1-ylmethyl) benzyl) -6-chloro-7V- (4-methoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amin at 5e 6c ^ 1 ^ ^NH l · JN CI ^ N ^NN 6c 6-Chlorine-7V- (3-methoxybenzyl) -1- (4- (piperidin-l-yl
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15/97 methyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-ami na 6c
THE
cr n
9f
6-Chloro-1- (3-fluoro-4- (pyrrolidin-1-ylmethyl) benzyl) -N- (4-methoxybenzyl) -IH-pyrazolo [3,4-d] pi rim idin-4-amine 9f [024] In another aspect, the present invention relates to a compound of formula (IE):
HN — W or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, where:
W is an amino protecting group, preferably tert-butoxycarbonyl, acetyl, benzyl, alii or p-methoxybenzyl;
the ring A, G, X ¹ , L ¹ - L ² , R ² ~ R ^{3 and} n are as defined in formula (I).
The compound of formula (I-E) includes, but is not limited to
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Example No. Structure and name of the compound If j] 1 N__ / 1f 6-Butoxy-N- (4-methoxybenzyl) -1- (4- (pyrrolidin-1ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-a mine If 2g pp _NH N ^ rX _M _ _ ii J, ^{N NN} v / = Π 2g 1- (4- (Azetidin-1-ylmethyl) benzyl) -6-butoxy-N- (4methoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amina 2g 3d pp _NH / 1 ^N NN ( 3d 6-Butoxy-N- (4-methoxybenzyl) -1- (4- (piperidin-1-i Imethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 3d 4c rj ^ X ^ NH Λ X ' ^N 4c 6-Butoxy-N- (3-methoxybenzyl) -1- (4- (pyrrolidin-1ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-a mine 4c
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5f _ _ II JN. N - ^N 'O 5f 1- (3- (Azetidin-1-ylmethyl) benzyl) - 6-butoxy-7V- (4methoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-ami na 5f 6d p ^ NHΧ ^ Γθ6d 6-Butoxy-7V- (3-methoxybenzyl) -1- (4- (piperidin-1-imethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 6d 7th rj ^ X ^ NH N'Vl wX o XXX 7th N- (4-Methoxybenzyl) -6- (2-methoxyethoxy) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyridinin-4-amine 7a 8th P ^ NH II JN ON ^N __ f) Λ XX- ^N 8a N- (4-Methoxybenzyl) -6 - (((1-methoxypropan-2-yl) oxy) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin- 4-amine 8a
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9g 1! JL, ^N 6-Butoxy-l- (3-fluoro-4- (pyrrolidin-1-ylmethyl) be nzil) -N- (4-methoxybenzyl) -IH-pyrazolo [3,4-d] pi midi-4-amine 9g 10th pp _NH ji / 1 - ^N / x N ^x N __ f ^H 10a Ab-Butyl-N ⁴ - (4-methoxybenzyl) -1- (4- (pyrrolidin-lilmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidine-4,6-diamine 10a 11a rj ^ h ^ 'NH OA JL' ^N íT ^NN 7 ^ O o if 11a Methyl 4 - ((4-methoxybenzyl) amino) -1- (4- (pyrrolidi n-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin a-6-carboxylate 11a 11b H | ^ Y% ⁰ 11 4 - (((4-Methoxybenzyl) amino) -N-propyl-1- (4- (pyrroleidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidine-6-carboxamide 11b 12b pp _NH Ji JL ' ^{N 0} 12b
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1- (4 - ((4-Methoxybenzyl) amino) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-6yl) pentan-l-one 12b
[025] In another aspect, the present invention relates to a method for preparing the compound of formula (I-E), comprising a step of:

subjecting a compound of formula (I-C) and a compound of formula (I-D) to a nucleophilic substitution reaction under an alkaline condition to obtain the compound of formula (I-E);
on what:
W is an amino protecting group, preferably tert-butoxycarbonyl, acetyl, benzyl, alii or p-methoxybenzyl;
X is halogen, preferably chlorine;
the ring A, G, VL ² , X ¹ , R ⁴ ~ R ^{3 and} n are as defined in the formula (IE).
[026] In another aspect, the present invention relates to a method for preparing the compound of formula (I), comprising a step of:
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removing the protecting group from the compound of formula (I-E) under an acidic condition to obtain the compound of formula (I);
on what:
W is an amino protecting group, preferably tert-butoxycarbonyl, acetyl, benzyl, alii or p-methoxybenzyl;
the ring A, G, L ¹ ~ L ² , X ¹ , Η ¹ - !! ^{3 and} n are as defined in the formula (D.
[027] In another aspect, the present invention relates to a compound of formula (II-B):
or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, where:
W is an amino protecting group, preferably
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21/97 tert-butoxycarbonyl, acetyl, benzyl, alii or p-methoxybenzyl;
X is halogen, preferably chlorine;
G, L ² , R ² , R ⁶ ~ R ^{7 and} n are as defined in formula (II).
[028] In another aspect, the present invention relates to a compound of formula (II-C):
or a tautomer, mesomer, racemate, enanthromer, diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof.
on what:
W is a protecting group: tert-butoxycarbonyl, acetyl,
G, L ¹ - L ² , R ² ~ R ² , R ^s ~ R ⁷ and (II).
[029] In another aspect, a method for preparing the comprising a step of:
t amino, preferably benzyl, alii or p-methoxybenzyl; n are as defined in the formula the present invention relates to a compound of formula (II-C),
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subjecting a compound of formula (II-B) and a compound of formula (I-D) to a nucleophilic substitution reaction under an alkaline condition to obtain the compound of formula (II-C);
on what:
W is an amino protecting group, preferably tert-butoxycarbonyl, acetyl, benzyl, allyl or p-methoxybenzyl;
X is halogen, preferably chlorine;
G, L ¹ - L ² , R ⁴ ~ R ² , R ^s ~ R ^{7 and} n are as defined in formula (II).
[030] In another aspect, the present invention relates to a method for preparing the compound of formula (II), comprising a step of:
remove the compound protecting group of formula (II-C)
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23/97 under an acidic condition to obtain the compound of formula (II);
on what:
W is an amino protecting group, preferably tert-butoxycarbonyl benzyl, allyl or p-methoxybenzyl;
G, L ¹ L ² , R ² ~ R ²
R ⁶ ~ R ⁷ en are as defined in the formula
In another aspect the present invention relates to a method for preparing the compound of formula (III) comprising a step of:
R1

removing the protecting group from the compound of formula (III-C) under an acidic condition to obtain the compound of formula (III);
on what:
W is an amino protecting group, preferably tert-butoxycarbonyl, acetyl, benzyl, allyl or p-methoxybenzyl;
L ¹ , R ² ~ R ² , are as defined in formula (III).
[032] In another aspect, the present invention relates to a pharmaceutical composition, comprising a therapeutically effective amount of the compound of formula (I), or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or mixtures thereof, or a pharmaceutically salt
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24/97 of the same and one or more pharmaceutically acceptable carriers, diluents or excipients.
[033] This invention also relates to a use of the compound of formula (I), or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, or pharmaceutical composition comprising the same, in the preparation of a medicament for TLR7 activation.
[034] The present invention also relates to a use of the compound of formula (I), or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising the same, in the preparation of a medicine for the treatment of an infection caused by a virus selected from the group consisting of dengue virus, yellow fever virus, West Nile virus, Japanese encephalitis virus, tick-borne encephalitis virus , Kunjin virus, Murray Valley encephalitis virus, St. Louis encephalitis virus, Omsk hemorrhagic fever virus, bovine viral diarrhea virus, Zika virus, HIV, HBV, HCV, HPV, RSV, SARS and influenza.
[035] The present invention also relates to a use of the compound of formula (I), or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof, or the composition
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25/97 comprising the same, in the preparation of a medicament for the treatment or prevention of melanoma, non-small cell lung carcinoma, hepatocellular carcinoma, basal cell carcinoma, renal cell carcinoma, myeloma, allergic rhinitis, asthma, COPD, ulcerative colitis and liver fibrosis.
[036] The present invention also relates to the compound of formula (I), or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same , for use as a medicine.
[037] The present invention also relates to the compound of formula (I), or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising the same , for use in TLR7 activation.
[038] The present invention also relates to the compound of formula (I), or a tautomer, mesomer, racemate, enantiomer, diastereoisomer or mixture thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising them, for use in the treatment of an infection caused by a virus selected from the group consisting of dengue virus, yellow fever virus,
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26/97 West Nile virus, Japanese encephalitis virus, tick-borne encephalitis virus, Kunjin virus, Murray Valley encephalitis virus, St. Louis encephalitis virus, Omsk hemorrhagic fever virus, bovine viral diarrhea virus , Zika virus, HIV, HBV, HCV, HPV virus, RSV, SARS and influenza.
[039] The present invention also relates to the compound of formula (I), or a tautomer, mesomer, racemate, enantiomer, diastereoisomer or mixture thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising them, for use in the treatment or prevention of melanoma, non-small cell lung carcinoma, hepatocellular carcinoma, basal cell carcinoma, renal cell carcinoma, myeloma, allergic rhinitis, asthma, COPD, ulcerative colitis and liver fibrosis.
[040] The present invention further relates to a method for activating TLR7, comprising administering to a patient in need thereof a therapeutically effective amount of the compound of formula (I) of the present invention, or a tautomer, mesomer, racemate, enantiomer, diastereoisomer , or a mixture thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising them.
[041] The present invention also refers to a method for treating an infection caused by a virus selected from the group consisting of dengue virus, yellow fever virus, virus
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West Nile 27/97, Japanese encephalitis virus, tick-borne encephalitis virus, Kunjin virus, Murray Valley encephalitis virus, St. Louis encephalitis virus, Omsk hemorrhagic fever virus, bovine viral diarrhea virus, virus Zika, HIV, HBV, HCV, HPV, RSV, SARS and influenza, comprising administering to a patient in need of a therapeutically effective amount of the compound, formula (I) of the present invention, or a tautomer, mesomer, racemate , enantiomer, diastereoisomer thereof, or mixtures thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising them.
[042] The present invention also relates to a method to treat or prevent melanoma, non-small cell lung carcinoma, hepatocellular carcinoma, basal cell carcinoma, renal cell carcinoma, myeloma, allergic rhinitis, asthma, COPD, ulcerative colitis and hepatic fibrosis, administration to a patient in need thereof a therapeutically effective amount of the compound of formula (I) of the present invention, or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a pharmaceutically salt acceptable content thereof, or the pharmaceutical composition comprising the same.
[043] Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral administration, for example, a tablet, troches, lozenge, aqueous suspension
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28/97 or oily, dispersible powder or granule, emulsion, hard or soft capsule or syrup or elixir. Oral compositions can be prepared according to any method known in the art for the preparation of pharmaceutical composition. Such a composition may contain one or more ingredients selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preservatives, in order to provide a pleasant and tasty pharmaceutical preparation. The tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets.
[044] An aqueous suspension contains the active ingredient in admixture with excipients suitable for the manufacture of an aqueous suspension. The aqueous suspension may also contain one or more preservatives, such as ethyl or n-propylparaben, one or more coloring agents, one or more flavoring agents and one or more sweetening agents.
[045] An oil suspension can be formulated by suspending the active ingredient in a vegetable oil. The oil suspension may contain a thickener. The aforementioned sweetening and flavoring agents can be added to provide a tasty formulation.
[046] The active ingredient in admixture with dispersants or wetting agents, suspending agent or one or more preservatives can be prepared as a dispersible powder or
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29/97 granule suitable for the preparation of an aqueous suspension by the addition of water. Suitable dispersants or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, such as sweetening agents, flavoring agents and coloring agents, can also be added. These compositions can be preserved by adding an antioxidant such as ascorbic acid.
[047] The pharmaceutical composition of the present invention can also be in the form of an oil-in-water emulsion.
[048] The pharmaceutical composition may be in the form of a sterile aqueous solution for injection. Acceptable vehicles and solvents that can be used are water, Ringer's solution and isotonic sodium chloride solution. The sterile injectable formulation can be a microemulsion of oil in sterile injectable water in which the active ingredient is dissolved in the oil phase. For example, the active ingredient is dissolved in a mixture of soy oil and lecithin, the oil solution is then added to a mixture of water and glycerol and processed to form a microemulsion. The solution for injection or microemulsion can be injected into the patient's bloodstream by local injection of embolus. Alternatively, it may be advantageous to administer the solution and the microemulsion in order to maintain a constant circulating concentration of the compound of the present invention. In order to maintain such a constant concentration, a device for continuous intravenous administration can be used. An example
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30/97 of such a device is the Deltec CADD-PLUS. ™. 5400 intravenous injection pump.
[049] The pharmaceutical composition can be in the form of a sterile injectable aqueous or oily suspension for intramuscular and subcutaneous administration. Such a suspension can be formulated with suitable dispersants or wetting agents and suspending agents, as described above, according to known techniques. The sterile injectable formulation may also be a sterile injectable solution or suspension prepared in an acceptable parenterally acceptable diluent or solvent. In addition, sterile fixed oils can easily be used as a solvent or suspending medium.
[050] The compound of the present invention can be administered in the form of a suppository for rectal administration. These pharmaceutical compositions can be prepared by mixing the appropriate common non-irritating drug that is solid at normal temperature but liquid in the rectum, thus melting in the rectum to release the drug. Such materials include cocoa butter, glycerin gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols with various molecular weights and polyethylene glycol fatty acid esters.
[051] It is well known to those skilled in the art that the dosage of a drug depends on a variety of factors including, but not limited to, the following factors: activity of a specific compound, patient age, weight
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31/97 of the patient, patient's general health, patient's behavior, patient's diet, time of administration, route of administration, rate of excretion, combination of drugs and the like. In addition, the ideal treatment, such as method of treatment, daily dose of the compound of formula (I) or the type of pharmaceutically acceptable salt thereof can be verified by conventional therapeutic regimens.
DETAILED DESCRIPTION OF THE INVENTION [052] Unless otherwise stated, the terms used in the specification and claims have the meanings described below.
[0 53] The term alkyl refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group comprising 1 to 20 carbon atoms, preferably an alkyl having 1 to 12 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl,
1.2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 1,2-trimethylpropyl, 1,1-dimethylbutyl, 1, 2-dimethylbutyl,
2.2- dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl,
2.3- dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl,
2.4- dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl,
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32/97
2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl,
2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl,
2.2- diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexile various branched isomers thereof. More preferably, an alkyl group is a lower alkyl having 1 to 6 carbon atoms and non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl,
1.3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group can be substituted or unsubstituted. When substituted, the substituent group (s) can be substituted at any available attachment point. The substituent group (s) is (are) preferably one or more groups independently selected optionally from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxy, nitro, cyano , cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heteroalkoxy, cycloalkylthio, heterocyclylthio, oxo, -OR ⁵ ,
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33/97
-C (O) R ⁵ , -S (O) _m R ⁵ , -NR ^S R ⁷ and -C (O) NR ⁶ R ⁷ .
[054] The term alkylene refers to a saturated, linear or branched aliphatic hydrocarbon group, having two residues derived from the removal of two hydrogen atoms from the same carbon atom or two different carbon atoms from the parent alkane. Linear or branched alkylene has 1 to 20 carbon atoms, preferably 1 to 12 carbon atoms, and more preferably 1 to 6 carbon atoms. Non-limiting examples of alkylene groups include, but are not limited to, methylene (-CH2-), 1,1-ethylene (-ΟΗ (ΟΗ ₃ ) -), 1,2-ethylene (-CH2CH2) -, 1, 1 -propylene (-CH (CH2CH3) -), 1,2-propylene (-CH2CH (CH ₃ ) -), 1,3-propylene (-CH ₂ CH ₂ CH ₂ -), 1,4-Butylene (-CH2CH2CH2CH2 -), and the like. The alkylene group can be substituted or unsubstituted. When substituted, the substituent group (s) can be substituted at any available attachment point. The substituent group (s) is (are) preferably one or more groups independently selected optionally from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxy, nitro, cyano , cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heteroalkoxy, cycloalkylthio,
heterocyclylthio, oxo, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ^S R ⁷ and -C (0) NR ^S R ⁷ . [055] 0 term alkenyl if refers to a group formed hydrocarbon by removing one or more atoms in
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34/97 hydrogen in an olefin molecule. The alkenyl group can be substituted or unsubstituted. When substituted, the substituent group (s) is (are) preferably one or more groups independently selected from the group consisting of hydrogen, alkyl, alkoxy, halogen, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro , cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ^S R ⁷ and -C (O) NR ⁶ R ⁷ .
[056] The term alkynyl refers to a hydrocarbon group containing a carbon-carbon triple bond in the molecule. The alkynyl group can be substituted or unsubstituted. When substituted, the substituent group (s) is (are) preferably one or more groups independently selected from the group consisting of hydrogen, alkyl, alkoxy, halogen, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl , heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ^S R ⁷ and -C (O) NR ⁶ R ⁷ .
[057] The term cycloalkyl refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon group having 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, preferably 3 to 10 carbon atoms, and more preferably 3 to 6 atoms of carbon. Non-limiting examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl and the like. Cycloalkyl
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Polycyclic 35/97 includes a cycloalkyl having a spiro ring, fused ring or bridged ring.
[058] The term amino protecting group refers to a group that prevents an amino group from reacting when other parts of the molecule are subject to a reaction, and can be easily removed. Non-limiting examples include tert-butoxycarbonyl, acetyl, benzyl, alii and p-methoxybenzyl and the like. These groups can be optionally substituted by one to three substituent groups selected from the group consisting of halogen, alkoxy and nitro. The amino protecting group is preferably p-methoxybenzyl.
[059] The term heterocyclyl refers to a 3 to 20 membered saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent group, where one or more ring atoms are hetero atoms selected from the group consisting of N, O and S (O ) _m (where m is an integer from 0 to 2), but excluding -0-0 -, - OS- or -SS- in the ring, the remaining atoms in the ring being carbon atoms. Preferably, the heterocyclyl has 3 to 12 ring atoms, where 1 to 4 atoms are hetero atoms, more preferably 3 to 10 ring atoms, where 1 to 4 atoms are hetero atoms and, more preferably, 5 to 6 ring atoms , where 1 to 3 atoms are hetero atoms. Non-limiting examples of monocyclic heterocyclyl include pyrrolidinyl, tetrahydropyranyl, 1,2,3,6-tetrahydropyridyl, piperidinyl, piperazinyl,
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36/97 morpholinyl, thiomorpholinyl, homopiperazinyl and the like. The polycyclic heterocyclyl includes a heterocyclyl with a spiro ring, fused ring or bridged ring.
[060] The heterocyclic ring can be fused to the aryl, heteroaryl or cycloalkyl ring, where the ring attached to the parent structure is heterocyclic. Non-limiting examples include:
[061] Heterocyclyl can be optionally substituted or unsubstituted. When substituted, the substituent group (s) is (are) preferably one or more group (s) independently optionally selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heteroalkoxy, cycloalkylthio, heterocyclylthio, oxo, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , - NR ^S R ⁷ and -C (O) NR ⁶ R ⁷ .
[062] The term aryl refers to a 6- to 14-membered all-carbon monocyclic ring or fused polycyclic ring (ie, each ring in the system shares an adjacent pair of carbon atoms with another ring in the system) having a system conjugate of n electrons, preferably 6 to 10 membered aryl, for example, phenyl and naphthyl. The aryl ring can be fused to the ring
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37/97 of heteroaryl, heterocyclyl or cycloalkyl, wherein the ring attached to the parent structure is the aryl ring. Non-limiting examples include:
[063] Aryl can be substituted or unsubstituted. When substituted, the substituent group (s) is (are) preferably one or more group (s) independently optionally selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heteroalkoxy, cycloalkylthio, heterocyclylthio, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ .
[064] The term heteroaryl refers to a 5 to 14 membered heteroaromatic system having 1 to 4 heteroatoms selected from the group consisting of O, S and N. The heteroaryl is preferably 5 to 10 membered heteroaryl, more preferably heteroaryl with 5 or 6 members, for example, furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, pyrazolyl,
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38/97 tetrazolyl and the like. The heteroaryl ring can be fused to the aryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is the heteroaryl ring. Non-limiting examples include:

[065] Heteroaryl can be optionally substituted or unsubstituted. When substituted, the substituent group (s) is (are) preferably one or more group (s) independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen , thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heteroalkoxy, cycloalkylthio, heterocyclylthio, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ .
[066] The term alkoxy refers to an O-(alkyl) group
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39/97 or an O-group (unsubstituted cycloalkyl), where alkyl is as defined above. Non-limiting examples of alkoxy include methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. The alkoxy can be optionally substituted or unsubstituted. When substituted, the substituent group (s) is (are) preferably one or more group (s) independently selected from the group consisting of halogen, alkyl, alkoxy, haloalkyl, hydroxyl, hydroxyalkyl, cyano , amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.
[067] The term haloalkyl refers to an alkyl group substituted by one or more halogens, where alkyl is as defined above. The term hydroxy refers to an -OH group.
[068] The term hydroxyalkyl refers to an alkyl group substituted by hydroxy (s), where alkyl is as defined above.
[069] The term halogen refers to fluorine, chlorine, bromine or iodine.
070] 0 term amino if refers The one group -nh ₂ . 071] 0 term cyan if refers The one group -CN. 072] 0 term nitro if refers The one group -no ₂ . 073] 0 term oxo if refers to wow i group = 0.
[074] Optionally or optionally means that the event or circumstance described subsequently may, but not necessarily, occur, and such description includes the situation in
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40/97 which event or circumstance occurs or not. For example, the heterocyclyl optionally substituted by an alkyl means that an alkyl group can be, but need not be present, and such a description includes the situation of the heterocyclyl being replaced by an alkyl and the heterocyclyl not being substituted by an alkyl.
[075] Substituted refers to one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, independently replaced by a corresponding number of substituents. It goes without saying that the substituents only exist in their possible chemical position. The person skilled in the art is able to determine whether substitution is possible or impossible by experiment or theory without paying excessive effort. For example, the combination of amino or hydroxy having free hydrogen and carbon atoms having unsaturated bonds (such as olefinic) can be unstable.
[076] A pharmaceutical composition refers to a mixture of one or more of the compounds described herein or physiologically / pharmaceutically acceptable salts or prodrugs with other chemical components, and other components such as physiologically / pharmaceutically acceptable carriers and excipients. The object of the pharmaceutical composition is to facilitate the administration of a compound to an organism, which is conducive to the absorption of
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41/97 active ingredient in order to show biological activity.
[077] A pharmaceutically acceptable salt refers to a salt of the compound of the present invention, which is safe and effective in mammals and has the desired biological activity.
[078] m and R ⁵ to R ⁷ are as defined in the compound of formula (D.
Synthesis Method of the Compound of the Present Invention [079] In order to achieve the object of the present invention, the present invention employs the following technical solutions:
[080] Scheme I [081] A method for preparing the compound of formula (I) of the present invention or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, comprises the following steps:
in the first step, a compound of formula (I-A) and a compound of formula (I-B) are subjected to a nucleophilic substitution reaction under an alkaline condition to obtain a compound of formula (I-C);
in the second step, the compound of formula (I-C) and a compound
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42/97 of formula (I-D) is subjected to a nucleophilic substitution reaction under an alkaline condition to obtain a compound of formula (I-E);
in the third step, the protecting group of the compound of formula (I-E) is removed under an acidic condition to obtain the compound of formula (I);
on what:
M is hydrogen or a metal ion, where the metal ion is preferably sodium ion;
W is an amino protecting group, preferably tert-butoxycarbonyl, acetyl, benzyl, alii or p-methoxybenzyl;
X is halogen, preferably chlorine;
the ring A, G, L ¹ ~ L ² , X ¹ , R ³ ~ R ^{3 and} n are as defined in the formula (D.
[082] The reagent that provides an alkaline condition includes organic bases and inorganic bases. Organic bases include, but are not limited to, triethylamine, N, .N-diisopropylethylamine, n-butyl lithium, lithium diisopropylamide, bis (trimethylsilyl) amine, potassium acetate, sodium tert-butoxide, tert-butoxide potassium and sodium n-butoxide. Inorganic bases include, but are not limited to, sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide.
[083] The reagent that provides an acidic condition includes,
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43/97 but is not limited to, hydrogen chloride, a solution of hydrogen chloride in 1,4-dioxane, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid, phosphoric acid, p- toluenesulfonic, MesSiCl, TMSOTf.
[084] The above reactions are preferably carried out in a solvent. The solvent used includes, but is not limited to, acetic acid, methanol, ethanol, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1,4-dioxane, water and N, Ndimethylformamide.
[085] Scheme II [086] A method for preparing the compound of formula (II) of the present invention or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof, comprises the following steps:
in the first step, a compound of formula (I-A) and a compound of formula (II-A) are subjected to a nucleophilic substitution reaction under an alkaline condition to obtain a compound
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44/97 of formula (II-B);
in the second step, the compound of formula (II-B) and a compound of formula (I-D) are subjected to a nucleophilic substitution reaction under an alkaline condition to obtain a compound of formula (11-C);
in the third step, the protecting group of the compound of formula (II-C) is removed under an acidic condition to obtain the compound of formula (II);
on what:
M is hydrogen or a metal ion, where the metal ion is preferably sodium ion;
W is an amino protecting group, preferably tert-butoxycarbonyl, acetyl, benzyl, alii or p-methoxybenzyl;
X is halogen, preferably chlorine;
G, L ¹ ~ L ² , R ² ~ R ² , R ^s ~ R ^{7 and} n are as defined in formula (II).
[087] The reagent that provides an alkaline condition includes organic bases and inorganic bases. Organic bases include, but are not limited to, triethylamine, N, N-diisopropylethylamine, n-butyl lithium, lithium diisopropylamide, bis (trimethylsilyl) amine, potassium acetate, sodium tert-butoxide, tert-butoxide potassium and sodium n-butoxide. Inorganic bases include, but are not limited to, sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide.
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45/97 [088] The reagent that provides an acidic condition includes, but is not limited to, hydrogen chloride, a solution of hydrogen chloride in 1,4-dioxane, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acids, methanesulfonic acid, nitric acid, phosphoric acid, p-toluenesulfonic acid, MesSiCl, TMSOTf.
[089] The above reactions are preferably carried out in a solvent. The solvent used includes, but is not limited to, acetic acid, methanol, ethanol, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1,4-dioxane, water and N, Ndimethylformamide.
[090] Scheme III [091] A method for preparing the compound of formula (III) of the present invention or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, comprises the following steps:
in the first step, a compound of formula (I-A) and a compound of formula (III-A) are subjected to a substitution reaction
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46/97 nucleophilic under an alkaline condition to obtain a compound of formula (III-B);
in the second step, the compound of formula (III-B) and a compound of formula (I-D) are subjected to a nucleophilic substitution reaction under an alkaline condition to obtain a compound of formula (111 - C);
in the third step, the protecting group of the compound of formula (III-C) is removed under an acidic condition to obtain the compound of formula (III);
on what:
M is hydrogen or a metal ion, where the metal ion is preferably sodium ion;
W is an amino protecting group, preferably tert-butoxycarbonyl, acetyl, benzyl, alii or p-methoxybenzyl;
X is halogen, preferably chlorine;
L ¹ , R ¹ ~ R ² , are as defined in formula (III).
[092] The reagent that provides an alkaline condition includes organic bases and inorganic bases. Organic bases include, but are not limited to, triethylamine, N, N-diisopropylethylamine, n-butyl lithium, lithium diisopropylamide, lithium bis (trimethylsilyl) amine, potassium acetate, sodium tert-butoxide, tert - potassium butoxide and sodium n-butoxide. Inorganic bases include, but are not limited to, sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, potassium acetate, cesium carbonate,
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47/97 sodium hydroxide and lithium hydroxide.
[093] The reagent that provides an acidic condition includes, but is not limited to, hydrogen chloride, a solution of hydrogen chloride in 1,4-dioxane, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acids, methanesulfonic acid, nitric acid, phosphoric acid, p-toluenesulfonic acid, MesSiCl, TMSOTf.
[094] The above reactions are preferably carried out in a solvent. The solvent used includes, but is not limited to, acetic acid, methanol, ethanol, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1,4-dioxane, water and N, Ndimethylformamide.
PREFERENTIAL MODALITIES [095] The present invention will be further described with reference to the following examples, but the examples are not to be considered as limiting the scope of the present invention.
EXAMPLES [096] The structures of the compounds were identified by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (õ) are displayed at 10 “ ⁶ (ppm). NMR was determined by a Bruker AVANCE-400 instrument. The solvents for the determination were deuterated dimethyl sulfoxide (DMSO-ds), deuterated chloroform (CDCls) and deuterated methanol (CD3OD), and the internal standard was tetramethylsilane (TMS).
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48/97 [097] The MS was determined by a FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Thermo, type: Finnigan LCQ advantage MAX).
[098] High performance liquid chromatography (HPLC) was determined in HPLC Agilent 1200DAD high pressure liquid chromatographs, Agilent 1200VWD HPLC and Waters HPLC e2695-2489.
[099] Chiral HPLC was determined on an Agilent HPLC 1260 DAD high performance liquid chromatograph.
[0100] The high performance liquid preparation was performed on the Waters 2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP and Gilson-281 chromatographs.
[0101] The chiral preparation was performed on a preparative chromatograph Shimadzu LC-20AP.
[0102] The CombiFlash rapid preparation instrument used was the Combiflash Rf200 (TELEDYNE ISCO).
[0103] The Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate was used as the thin layer silica gel (TLC) chromatography plate. The size of the silica gel plate used in TLC was 0.15 mm to 0.2 mm and the size of the silica gel plate used in the purification of the product was 0.4 mm to 0.5 mm.
[0104] 200 to 300 mesh silica gel from Yantai Huanghai was generally used as a carrier for column chromatography.
[0105] Average kinase inhibition rates and values
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49/97 of IC50 were determined by a NOVOSTAR ELISA (BMG Co., Germany).
[0106] The known starting materials of the present invention can be prepared by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organnics, Aldrich Chemical Company, Accela ChemBio Inc., or Dari chemical Company, etc. .
[0107] Unless otherwise specified, reactions were carried out under an argon atmosphere or a nitrogen atmosphere.
[0108] Argon atmosphere or nitrogen atmosphere means that a reaction flask is equipped with an argon or nitrogen flask (about 1 L).
[0109] Hydrogen atmosphere means that a reaction flask is equipped with a hydrogen flask (about 1 L).
[0110] Pressurized hydrogenation reactions were performed on a Parr 3916EKX hydrogenation instrument and on a Qinglan QL-500 hydrogen generator or on an HC2-SS hydrogenation instrument.
[0111] In hydrogenation reactions, the reaction system was generally aspirated and filled with hydrogen, with the above operation being repeated three times.
[0112] The CEM Discover-S 908860 microwave reactor was used in microwave reactions.
[0113] Unless otherwise indicated, the solution refers to an aqueous solution.
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50/97 [0114] Unless otherwise specified, the reaction temperature is the ambient temperature between 20 ° C and 30 ° C.
[0115] The reaction process in the examples was monitored by thin layer chromatography (TLC). The development solvent used in the reactions, the chromatographic column elution system and the development solvent system in thin layer chromatography for purification of the compounds included: A: dichloromethane / methanol system and B: nhexane / ethyl acetate system. The volume ratio of the solvent has been adjusted according to the polarity of the compounds, and a small amount of alkaline reagent, such as triethylamine or acid reagent, such as acetic acid, can also be added for adjustment.
[0116] Example 1
6-Butoxy-l- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazole [3,4-d] pyrimidin-4-amine
NH.
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Step 1
6-Chloro-7V- (4-methoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin -4-amine lc [0117] 4,6-Dichloro-1H-pyrazolo [3,4-d] pyrimidine la ( 120 mg, 0.63 mmol), 4-methoxybenzylamine lb (87.1 mg, 0.63 mmol) and triethylamine (64.13 mg, 0.63 mmol) were dissolved in 2 mL of tetrahydrofuran and the reaction solution was stirred at room temperature for 1 hour. The reaction was stopped and the reaction solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with elution system A to obtain the title compound lc (140 mg, yield: 76.1%).
[0118] MS m / z (ESI): 290.2 [M + l]
Step 2
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6-Chloro-7V- (4-methoxybenzyl) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine le [0119] Compound lc (140 mg, 0.48 mmol),
1- (4- (chloromethyl) benzyl) pyrrolidine Id (101.34 mg, 0.48 mmol, prepared according to the procedure disclosed in patent application WC2002012224) and potassium carbonate (66.79 mg, 0.48 mmol ) were dissolved in 2 ml of N, N-dimethylformamide. The reaction was stopped after stirring at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel with elution system A to obtain the title compound 1 (70 mg, yield: 31.3%).
[0120] MS m / z (ESI): 463.2 [M + l]
Step 3
6-Butoxy-7V- (4-methoxybenzyl) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine lf [0121] The compound le ( 70 mg, 0.15 mmol), sodium n-butoxide (0.3 mL, 0.60 mmol) and 1 mL of n-butanol were added to a microwave tube successively, heated to 160 ° C and stirred for 1.5 hours. The reaction was stopped and the reaction solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with Aparaseobtero elution system composed of the title lf (40 mg, yield: 52.8%).
[0122] MS m / z (ESI): 501.2 [M + l]
Step 4
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6-Butoxy-l- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazole o [3,4-d] pyrimidin-4-amine 1 [0123] The If compound (40 mg, 0.08 mmol ) and 2 ml of trifluoroacetic acid were added to a reaction flask, heated to reflux and stirred for 24 hours. The reaction was stopped and the reaction solution was concentrated under reduced pressure and added with 1 ml of ammonia in methanol. The residue was purified by thin layer chromatography with solvent development system A to obtain the title compound 1 (15 mg, yield: 46.0%).
[0124] MS m / z (ESI): 381.2 [M + l]
NMR (400MHz, CD ₃ OD) δ 7.98 (s, 1H), 7.41 (d, 2H), 7.36 (d, 2H), 5.48 (s, 2H), 4.39 (t , 2H), 4.13 (s, 2H), 3.12-3.08 (m, 4H), 2.02-1, 98 (m, 4H), 1.80-1.76 (m, 2H ), 1.55-1.49 (m, 2H), 1.01 (t, 3H).
Example 2
1- (4- (Azetidin-1-ylmethyl) benzyl) -6-butoxy-1H-pyrazolo [
3,4-d] pyrimidin-4-amine 2
NH
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Step 1
Methyl 4- (azetidin-1-ylmethyl) benzoate 2c [0125] 4- (bromomethyl) methyl 2a benzoate (1.0 g, 4.37 mmol), azetidine 2b (299 mg, 5.24 mmol) and triethylamine ( 529 mg, 5.24 mmol) were dissolved in 10 ml of tetrahydrofuran and the reaction solution was stirred at room temperature for 16 hours. The reaction solution was added with water (100 ml) and extracted with ethyl acetate (100 ml). The organic phase was washed with saturated sodium chloride solution (100 ml), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain crude title compound 2c (880 mg), which was used directly in the next step without purification.
[0126] MS m / z (ESI): 206.1 [M + l]
Step 2
4- (Azetidin-1-ylmethyl) phenylcarbinol 2d
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55/97 [0127] Crude compound 2c (880 mg, 0.33 mmol) was dissolved in 10 ml of diethyl ether, aluminum and lithium hydride (326 mg, 8.57 mmol) was added at 0 ° C and the reaction solution was stirred at 0 ° C for 2 hours. 0.3 ml of water, 0.3 ml of 15% sodium hydroxide solution and 0.9 ml of water were added successively to quench the reaction. The reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain the crude title compound 2d (700 mg), which was used directly in the next step without purification.
[0128] MS m / z (ESI): 178.3 [M + l]
Step 3
1- (4- (Chloromethyl) benzyl) azetidine 2e [0129] The crude compound 2d (700 mg, 3.95 mmol) was dissolved in 10 ml of dichloromethane, thionyl chloride (0.58 ml, 7, 90 mmol) at 0 ° C and the reaction solution was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure and the resulting residue was added with saturated sodium carbonate solution (50 ml) and extracted with dichloromethane (100 ml x 2). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain crude title compound 2e (720 mg), which was used directly in the next step without purification.
[0130] MS m / z (ESI): 197.2 [M + l]
Step 4
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1- (4- (Azetidin-1-ylmethyl) benzyl) -6-chloro-N- (4-methoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 2f [0131] Compound lc ( 600 mg, 2.07 mmol), crude compound 2e (405 mg, 2.07 mmol) and potassium carbonate (286 mg, 2.07 mmol) were dissolved in 10 mL of N, N-dimethylformamide and the solution The reaction mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and the resulting residue was purified by column chromatography on silica gel with elution system A to obtain the title compound 2f (300 mg, yield: 32.3%).
[0132] MS m / z (ESI): 449.2 [M + l]
Step 5
1- (4- (Azetidin-1-ylmethyl) benzyl) -6-butoxy-N- (4-methoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 2g [0133] Compound 2f (150 mg, 0.33 mmol), sodium n-butoxide (0.7 mL, 1.40 mmol) and 2 mL of n-butanol were added to a microwave tube successively, heated to 160 ° C and stirred for 1.5 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with elution system A to obtain the title compound 2g (60 mg, yield: 36.9%).
[0134] MS m / z (ESI): 487.3 [M + l]
Step 6
1- (4- (Azetidin-1-ylmethyl) benzyl) -6-butoxy-1H-pyrazolo [
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3,4-d] pyrimidin-4-amine 2 [0135] Compound 2g (60 mg, 0.12 mmol) and 2 ml of trifluoroacetic acid were added to a reaction flask, heated to reflux and stirred for 24 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by high performance liquid chromatography (Waters-2767, elution system: 10 mmoL / L of ammonium bicarbonate, water, acetonitrile) to obtain the title compound 2 (15 mg, yield: 33.2 %).
[0136] MS m / z (ESI): 367.2 [M + 1] [0137] ⁴ H NMR (400MHz, CD ₃ OD) δ 7, 97 (s, 1H), 7.34-7.26 ( m,
4H), 5.44 (s, 2H), 4.39 (t, 2H), 3.77 (s, 2H), 3.47 (t, 4H), 2.22-2.18 (m, 2H ), 1.80-1.76 (m, 2H), 1.55-1.49 (m, 2H), 1.01 (t, 3H).
Example 3
6-Butoxy-l- (4- (piperidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 3
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Step 1
1- (4- (Chloromethyl) benzyl) piperidine 3b [0138] O 4- (Piperidin-1-ylmethyl) phenylcarbinol 3a (1.17 g, 5.70 mmol, prepared according to the known method disclosed in Journal of Medicinal Chemistry, 2003, 46 (8), 1523-1530) was dissolved in 20 ml of dichloromethane, thionyl chloride (0.83 ml, 11.4 mmol) was added at 0 ° C and the reaction solution was stirred at temperature room for 3 hours. The reaction solution was warmed to room temperature and concentrated under reduced pressure. The reaction mixture was added with saturated sodium carbonate solution (50 mL) and extracted with dichloromethane (100 mL χ 2). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain the crude title compound 3b (1.2 g), which was used directly in the next step without purification.
MS m / z (ESI): 224.2 [M + l]
Step 2
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6-Chloro-7V- (4-methoxybenzyl) -1- (4- (piperidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 3c [0139] Compound lc ( 1.5 g, 5.18 mmol), crude compound 3b (1.16 g, 5.18 mmol) and potassium carbonate (716 mg, 5.18 mmol) were dissolved in 20 mL of N, N- dimethylformamide and the reaction solution was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and the resulting residue was purified by column chromatography on silica gel with elution system A to obtain the title compound 3c (400 mg, yield: 16.2%).
[0140] MS m / z (ESI): 477.3 [M + l]
Step 3
6-Butoxy-7V- (4-methoxybenzyl) -1- (4- (piperidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 3d [0141] Compound 3c ( 100 mg, 0.21 mmol), sodium n-butoxide (0.2 mL, 0.80 mmol) and 1 mL of n-butanol were added to a microwave tube successively, heated to 160 ° C and stirred for 1.5 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with elution system A to obtain the title compound 3d (50 mg, yield: 46.3%).
[0142] MS m / z (ESI): 515.3 [M + l]
Step 4
6-Butoxy-l- (4- (piperidin-1-ylmethyl) benzyl) -IH-pyrazole
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60/97 [3,4-d] pyrimidin-4-amine 3 [0143] Compound 3d (60 mg, 0.12 mmol) and 2 mL of trifluoroacetic acid were added to a reaction flask. The reaction solution was heated to reflux and stirred for 24 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography with the development system to obtain the title compound 3 (20 mg, yield: 49.5%).
[0144] MS m / z (ESI): 395.3 [M + l] [0145] ^Χ Η NMR (400MHz, CD ₃ OD) δ 7, 99 (s, 1H), 7.47-7.38 ( m, 4H), 5.49 (s, 2H), 4.39 (t, 2H), 4.18 (s, 2H), 3.09-3.00 (m, 4H), 1.81-1 , 76 (m, 6H), 1.68-1, 62 (m, 2H), 1.55-1.49 (m, 2H), 1.00 (t, 3H).
Example 4
6-Butoxy-l- (3- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazole o [3,4-d] pyrimidin-4-amine 4
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4th
Step 1
6-Chloro-N- (3-methoxybenzyl) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 4b [0146] Compound lc (1 , 0 g, 3.45 mmol), 1- (3- (chloromethyl) benzyl) pyrrolidine 4a (724 mg, 3.45 mmol, prepared according to the method described in patent application W02016040419) and potassium carbonate (377 mg, 3.45 mmol) were dissolved in 10 ml of N, N-dimethylformamide. The reaction was stopped after stirring at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and the resulting residue was purified by column chromatography on silica gel with elution system A to obtain the title compound 4b (300 mg, yield: 18.7%).
[0147] MS m / z (ESI): 463.2 [M + l]
Step 2
6-Butoxy-N- (3-methoxybenzyl) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 4c [0148] Compound 4b ( 300 mg, 0.65 mmol), n-butoxide
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62 / 9Ί sodium (1.3 ml, 2.60 mmol) and 2 ml of n-butanol were added to a microwave tube successively, heated to 160 ° C and stirred for 1.5 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with elution system A to obtain the title compound 4c (140 mg, yield: 43.1%).
[0149] MS m / z (ESI): 501.2 [M + l]
Step 3
6-Butoxy-l- (3- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazole [3,4-d] pyrimidin-4-amine 4 [0150] Compound 4c (140 mg, 0.08 mmol ) and 2 ml of trifluoroacetic acid were added to a reaction flask, heated to reflux and stirred for 24 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by high-performance liquid chromatography (Waters-2767, elution system: 10 mmoL / L of ammonium bicarbonate, water, acetonitrile) to obtain the title compound 4 (60 mg, yield: 56.3 %).
[0151] MS m / z (ESI): 381.2 [M + l] [0152] NMR (400MHz, CD ₃ OD) δ 7.98 (s, 1H), 7.35-725 (m, 4H) , 5.47 (s, 2H), 4.39 (t, 2H), 3.81 (s, 2H), 2.76-2.70 (m, 4H), 1.98-1, 93 (m , 4H), 1.79-1.76 (m, 2H), 1.55-1.50 (m, 2H),
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1.01 (t, 3H).
Example 5
1- (3- (Azetidin-1-ylmethyl) benzyl) -6-butoxy-1H-pyrazolo [
3,4-d] pyrimidin-4-amine 5


Step 1
Methyl 3- (azetidin-1-ylmethyl) benzoate 5b [0153] 3- (bromomethyl) methyl benzoate 5a (1.0 g, 4.37 mmol), azetidine 2b (299 mg, 5.24 mmol) and triethylamine ( 529 mg, 5.24 mmol) were dissolved in 10 ml of tetrahydrofuran and the reaction solution was stirred at room temperature for 16 hours. The reaction solution was added with water (100 ml) and extracted with ethyl acetate (100 ml). The organic phase was washed with saturated sodium chloride solution (100 ml), dried
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[0154] MS m / z (ESI): 206.1 [M + l]
Step 2
3- (Azetidin-1-ylmethyl) phenylcarbinol 5c [0155] Compound crude 5b (840 mg, 4.09 mmol) was dissolved in 10 ml of diethyl ether, lithium aluminum hydride (310 mg, 8, 19 mmol) at 0 ° C and stirred at 0 ° C for 2 hours. 0.3 ml of water, 0.3 ml of 15% sodium hydroxide solution and 0.9 ml of water were added successively to quench the reaction. The reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain the crude title compound 5c (700 mg), which was used directly in the next step without purification.
[0156] MS m / z (ESI): 178.3 [M + l]
Step 3
1- (3- (Chloromethyl) benzyl) azetidine 5d [0157] Crude compound 5c (700 mg, 3.95 mmol) was dissolved in 10 mL of dichloromethane, thionyl chloride (0.58 mL, 7, 90 mmol) at 0 ° C and stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, added with the saturated sodium carbonate solution (50 ml) and extracted with dichloromethane (100
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65/97 mL χ 2). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain crude title compound 5d (700 mg), which was used directly in the next step without purification.
[0158] MS m / z (ESI): 197.2 [M + l]
Step 4
1- (3- (Azetidin-1-ylmethyl) benzyl) -6-chloro-N- (4-methoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 5e [0159] Compound lc ( 300 mg, 1.04 mmol), crude compound 5d (203 mg, 1.04 mmol) and potassium carbonate (144 mg, 1.04 mmol) were dissolved in 5 mL of N, N-dimethylformamide and the solution The reaction mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and the resulting residue was purified by column chromatography on silica gel with elution system A to obtain the title compound 5e (30 mg, yield: 6.5%).
[0160] MS m / z (ESI): 449.2 [M + l]
Step 5
1- (3- (Azetidin-1-ylmethyl) benzyl) -6-butoxy-N- (4-methoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 5f [0161] Compound 5e (50 mg, 0.11 mmol), sodium n-butoxide (0.2 mL, 0.40 mmol) and 1 mL of n-butanol were added to a microwave tube successively, heated to 160 ° C and stirred for 1.5 hours. The reaction solution was cooled to
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[0162] MS m / z (ESI): 487.3 [M + l]
Step 6
1- (3- (Azetidin-1-ylmethyl) benzyl) -6-butoxy-1H-pyrazolo [
3,4-d] pyrimidin-4-amine 5 [0163] Compound 5f (35 mg, 0.07 mmol) and 1 ml of trifluoroacetic acid were added to a reaction flask. The reaction solution was heated to reflux and stirred for 24 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by high performance liquid chromatography (Waters-2767, elution system: 10 mmoL / L of ammonium bicarbonate, water, acetonitrile) to obtain the title compound 5 (2.0 mg, yield: 7 , 9%).
[0164] MS m / z (ESI): 367.2 [M + l] [0165] ^Χ Η NMR (400MHz, CD ₃ OD) δ 7, 98 (s, 1H), 7.30-7.28 ( m, 1H), 7.22-7.19 (m, 3H), 5.45 (s, 2H), 4.39 (t, 2H), 3.60 (s, 2H), 3.2 8 ( t, 4H), 2.12-2.09 (m, 2H), 1.80-1.76 (m, 2H), 1.55-1.49 (m, 2H), 1.00 (t, 3H).
Example 6
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6-Butoxy-l- (3- (piperidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 6
Step 1
1- (3- (Chloromethyl) benzyl) piperidine 6b [0166] 3- (Piperidin-1-ylmethyl) phenylcarbinol 6a (1.7 g,
8.28 mmol, prepared according to the known method disclosed in Bioorganic & Medicinal Chemistry, 2004, 12 (10), 2727-2736) was dissolved in 20 ml of dichloromethane, thionyl chloride (1.2 ml, 16.56 mmol) was added at 0 ° C and the reaction solution was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, added with the saturated sodium carbonate solution (50 ml) and extracted with dichloromethane (100 ml * 2). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain
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[0167] MS m / z (ESI): 224.2 [M + l]
Step 2
6-Chloro-7V- (3-methoxybenzyl) -1- (4- (piperidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 6c [0168] Compound lc ( 300 mg, 1.04 mmol), crude compound 6b (232 mg, 1.04 mmol) and potassium carbonate (144 mg, 1.04 mmol) were dissolved in 5 mL of N, N-dimethylformamide. The reaction was stopped after stirring at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and the resulting residue was purified by column chromatography on silica gel with elution system A to obtain the title compound 6c (50 mg, yield: 10.1%).
[0169] MS m / z (ESI): 477.3 [M + l]
Step 3
6-Butoxy-7V- (3-methoxybenzyl) -1- (4- (piperidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 6d [0170] Compound 6c ( 50 mg, 0.10 mmol), sodium n-butoxide (0.2 mL, 0.40 mmol) and 1 mL of n-butanol were added to a microwave tube successively, heated to 160 ° C and stirred for 1.5 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with elution system A to obtain the compound
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[0171] MS m / z (ESI): 515.3 [M + l]
Step 4
6-Butoxy-1- (3- (piperidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 6 [0172] Compound 6d (30 mg, 0.06 mmol) and 2 ml of trifluoroacetic acid were added to a reaction flask, heated to reflux and stirred for 24 hours. The reaction was stopped and the reaction solution was concentrated under reduced pressure and added with a solution of 7 N ammonia in methanol (1 mL). The reaction solution was concentrated under reduced pressure and the residue was purified by thin layer chromatography with development system A to obtain the title compound 6 (7.0 mg, yield: 29.2%).
[0173] MS m / z (ESI): 395.3 [M + l] [0174] X NMR (400MHz, CD ₃ OD) δ 7.99 (s, 1H), 738-7.31 (m, 4H ), 5.48 (s, 2H), 4.38 (t, 2H), 3.86 (s, 2H), 2.87-2.80 (m, 4H), 1.79-1.75 ( m, 2H), 1.71-1.68 (m, 4H), 1.54-1.40 (m, 4H), 1.00 (t, 3H).
Example 7
6- (2-Methoxyethoxy) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -1 H-pyrazolo [3,4-d] pyrimidin-4-amine 7
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Step 1
N- (4-Methoxybenzyl) -6- (2-methoxyethoxy) -1- (4- (pyrrolidin-
1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 7a [0175] The compound le (90 mg, 0.19 mmol), sodium 2-methoxyethanol (0.3 mL, 0.60 mmol) and 1 ml of 2-methoxyethanol were added to a microwave tube successively, heated to 160 ° C and stirred for 1.5 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with elution system A to obtain the title compound 7a (30 mg, yield: 30.7%).
[0176] MS m / z (ESI): 503.3 [M + l]
Step 2
6- (2-Methoxyethoxy) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -1 H-pyrazolo [3,4-d] pyrimidin-4-amine 7 [0177] Compound 7a (30 mg, 0.06 mmol) and 5 mL of trifluoroacetic acid were added to a reaction flask, heated to 100 ° C and stirred for 2 hours. The reaction solution
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/1 / 9Ί was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by high performance liquid chromatography (Waters-2767, elution system: 10 mmoL / L of ammonium bicarbonate, water, acetonitrile) to obtain the title compound 7 (5 mg, yield: 19.7 %).
[0178] MS m / z (ESI): 383.2 [M + l] [0179] ^X H NMR (400MHz, CD ₃ OD) δ 7, 96 (s, 1H), 7.30-7.28 (d, 2H), 7.25-7.23 (d, 2H), 5.42 (s, 2H), 4.51-4.48 (t, 2H), 3.74-3.72 (t, 2H) , 3.65 (s, 2H), 3.39 (s, 3H), 2.57 (s, 4H), 1.81 to 1.78 (m, 4H).
Example 8
6 - ((l-Methoxypropan-2-yl) oxy) -1- (4- (pyrrolidin-l-ylmethyl
1) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 8

Step 1
N- (4-Methoxybenzyl) -6 - (((1-methoxypropan-2-yl) oxy) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidin- 4
Petition 870190046717, of 5/20/2019, p. 87/130 / 97 amine 8a [0180] The compound le (200 mg, 0.43 mmol),
2-methoxy-1-methyl-ethoxy sodium (96.9 mg, 0.86 mmol) and 5 ml of propylene glycol methyl ether were added to a microwave tube successively, heated to 160 ° C and stirred for 1 , 5 hour. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with elution system A to obtain the title compound 8a (150 mg, yield: 67.2%).
[0181] MS m / z (ESI): 517.3 [M + l]
Step 2
6- (((l-Methoxypropan-2-yl) oxy) -1- (4- (pyrrolidin-l-ylmethyl
1) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 8 [0182] Compound 8a (80 mg, 0.15 mmol) and 5 ml of trifluoroacetic acid were added to a reaction flask, heated to 80 ° and stirred for 1 hour. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by high-performance liquid chromatography (Waters-2767, elution system: 10 mmoL / L ammonium bicarbonate, water, acetonitrile) to obtain the title compound 8 (20 mg, yield: 32.6 %).
[0183] MS m / z (ESI): 397.2 [M + l]
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73/97 [0184] ^X H NMR (400MHz, CD ₃ OD) δ 7, 95 (s, 1H), 7.35-7.33 (d, 2H), 7.29-7.27 (d, 2H ), 5.43 (m, 3H), 3.83 (s, 2H), 3.60-3.52 (m, 2H), 3.37 (s, 3H), 2.76 (s, 4H) , 1.87 (s, 4H), 1.34-1.32 (t, 3H).
Example 9
6-Butoxy-l- (3-fluoro-4- (pyrrolidin-1-ylmethyl) benzyl) -1
H-pyrazolo [3,4-d] pyrimidin-4-amine 9
Step 1
3-Fluoro-4- (pyrrolidin-1-ylmethyl) benzonitrile 9c
4- (Bromomethyl) -3-fluorobenzonitrile 9a (1 g, 4.67 mmol), pyrrolidine 9b (332 mg, 4.67 mmol) and N, N-diisopropylethylamine (1.21 g, 9.34 mmol) were dissolved in 10 ml of acetonitrile.
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After stirring for 2 hours, the reaction solution was concentrated under reduced pressure to obtain the crude title compound 9c (1 g), which was used directly in the next step without purification.
[0185] MS m / z (ESI): 205.4 [M + l]
Step 2 [0186] 3-Fluoro-4- (pyrrolidin-1-ylmethyl) benzoic acid 9d [0187] The crude compound 9c (1 g, 4.9 mmol) was dissolved in a mixed solvent of 5 mL of sulfuric acid, 5 ml of water and 10 ml of acetic acid. The reaction was stopped after stirring at 90 ° C for 16 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The residue was added with methanol and filtered to remove insoluble matter. The filtrate was concentrated under reduced pressure to obtain the crude title compound 9d (1 g), which was used directly in the next step without purification.
[0188] MS m / z (ESI): 224.4 [M + l]
Step 3 (3-Fluoro-4- (pyrrolidin-1-ylmethyl) phenyl) methanol 9e [0189] The crude compound 9d (1 g, 4.48 mmol) was dissolved in 20 ml of tetrahydrofuran. The reaction solution was cooled to 0 ° C, added with lithium aluminum hydride (607 mg, 17.9 mmol) and stirred for 3 hours. 1 ml of water, 1 ml of 2 N sodium hydroxide solution and 3 ml of water were added
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75/97 successively to quench the reaction. The reaction solution was filtered and the filtrate was collected and concentrated under reduced pressure to obtain the crude title compound 9e (820 mg), which was used directly in the next step without purification.
[0190] MS m / z (ESI): 210.4 [M + l]
Step 4
6-Chloro-1- (3-fluoro-4- (pyrrolidin-1-ylmethyl) benzyl) -N (4-methoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 9f [0191] O crude compound 9e (100 mg, 0.48 mmol), compound lc (141.34 mg, 0.48 mmol) and triphenylphosphine (192 mg, 0.73 mmol) were dissolved in 10 mL of 1,4-dioxane and diisopropyl azodicarboxylate (148 mg, 0.73 mmol) was then added dropwise. The reaction solution was heated to 85 ° C and stirred for 4 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with elution system A to obtain the title compound 9f (90 mg, yield: 38.3%).
[0192] MS m / z (ESI): 481.4 [M + l]
Step 5
6-Butoxy-1- (3-fluoro-4- (pyrrolidin-1-ylmethyl) benzyl) -N - (4-methoxybenzyl) -IH-pyrazolo [3,4-d] pyrimidin-4-amine 9g [0193] Compound 9f (90 mg, 0.19 mmol), sodium n-butoxide (18 mg, 0.18 mmol) and 5 mL of n-butanol were added to a
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Ί6 / 9Ί microwave tubes in succession, heated to 160 ° C and stirred for 1.5 hours. The reaction was stopped and the reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with elution system A to obtain the title compound 9g (35 mg, yield: 36.1%).
[0194] MS m / z (ESI): 519.5 [M + l]
Step 6
6-Butoxy-l- (3-fluoro-4- (pyrrolidin-1-ylmethyl) benzyl) -1
H-pyrazolo [3,4-d] pyrimidin-4-amine 9 [0195] Compound 9g (35 mg, 0.07 mmol) and 10 mL of trifluoroacetic acid were added to a sealed tube, heated to 100 ° C and stirred for 1 hour. The reaction was stopped and the reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by high-performance liquid chromatography (Waters-2767, elution system: 10 mmoL / L ammonium bicarbonate, water, acetonitrile) to obtain the title 9 compound (20 mg, yield: 74.3 %).
[0196] MS m / z (ESI): 399.5 [M + 1] [0197] ^X H NMR (400MHz, CD ₃ OD) δ 7, 97 (s, 1H), 7.37-7.33 ( m, 1H), 7, 07-6, 99 (m, 2H), 5.42 (s, 2H), 4.38-4.35 (t, 2H), 3.68 (s, 2H), 2 , 56 (s, 4H), 1.79-1, 73 (m, 6H), 1.52-1.46 (m, 2H), 0.99-0.96 (t, 3H).
Example 10
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7V ⁶ -Butyl-1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazole [3,4-d] pyrimidine-4,6-diamine 10

Step 1
Ab-Butyl-N ⁴ - (4-methoxybenzyl) -1- (4- (pyrrolidin-l-ylmethyl
1) benzyl) -IH-pyrazolo [3,4-d] pyrimidine-4,6-diamine 10a [0198] The compound le (50 mg, 0.11 mmol), n-butylamine (23.7 mg, 0, 32 mmol) and N, N-diisopropylethylamine (41, 9 mg, 0.32 mmol) were added to 5 ml of n-butanol successively. The reaction solution was heated to 120 ° C and stirred in a microwave for 1 hour. The reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain the crude title compound 10a (20 mg), which was used directly in the next step without purification.
Step 2
Ab-Butyl-l- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazole
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78/97 ο [3,4-d] pyrimidine-4,6-diamine 10 [0199] The crude compound 10a (20 mg, 0.04 mmol) and 5 mL of trifluoroacetic acid were added to a reaction flask, heated to 100 ° C and stirred overnight. The reaction was stopped and the reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by high performance liquid chromatography (Waters-2767, elution system: 10 mmoL / L of ammonium bicarbonate, water, acetonitrile) to obtain the title compound 10 (15.2 mg, a yellow solid , yield: 62.5%).
[0200] MS m / z (ESI): 380.3 [M + l] [0201] NMR (400MHz, CD ₃ OD) δ 8.04 (s, 1H), 7.49-7.47 (d, 2H), 7.42-7.40 (d, 2H), 5.44 (s, 2H), 4.34 (s, 2H), 3.49-3.45 (m, 4H), 3.15 (s, 2H), 2.13 (s, 2H), 1.93 (s, 2H), 1.65-1, 60 (m, 2H), 1.45-1.39 (m, 2H), 0.98-0.94 (t, 3H).
Example 11
4-Amino-7V-propyl-l- (4- (pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidine-6-carboxamide 11
NH ₂
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Ί9 / 9Ί

Step 1
Methyl
4 - ((4-methoxybenzyl) amino) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidine-6-carboxylate 11a [0202] The compound le (200 mg, 0.43mmol), palladium acetate (2.9 mg, 0.013 mmo1),
4,5-bisdiphenyl-phosphine-9,9-dimethyloxanthene (15mg, 0.026mmol) and triethylamine (44mg, 0.4mmol) were dissolved in 3mL of n-butanol and 3mL of N, N-dimethylformamide. The reaction system was purged with carbon monoxide three times. The reaction solution was heated to 70 ° C and stirred for 16 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with elution system A to obtain the title compound 11a (150 mg, yield: 71.4%).
[0203] MS m / z (ESI): 487.5 [M + l]
Step 2
4 - ((4-Methoxybenzyl) amino) -N-propyl-l- (4- (pyrrolidin-1
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80/97 ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidine-6-carboxamide lib [0204] Compound 11a (50 mg, 0.1 mmol) and n-propylamine (12 mg, 0.2 mmol) were dissolved in 5 ml of ethanol successively. The reaction solution was added to a sealed tube, heated to 60 ° C and stirred for 16 hours. The reaction was stopped and the reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain the crude title compound 11b (20 mg), which was used directly in the next step without purification.
Step 3
4-Amino-7V-propyl-l- (4- (pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidine-6-carboxamide 11 [0205] Crude compound 11b (20 mg , 0.04 mmol) and 5 mL of trifluoroacetic acid were added to a reaction flask, heated to 100 ° C and stirred for 12 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by high performance liquid chromatography (Waters-2767, elution system: 10 mmoL / L of ammonium bicarbonate, water, acetonitrile) to obtain the title compound 11 (10 mg, yield: 60.3 %).
[0206] MS m / z (ESI): 394.5 [M + l] [0207] NMR (400MHz, CD ₃ OD) δ 8.12 (s, 1H), 7.28 (m, 4H), 5 , 64 (s, 2H), 3.62 (s, 2H), 3.41-3.37 (t, 2H), 2.55-2.52 (m,
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4H), 1.80-1.77 (m, 4H), 1.70-1.64 (m, 2H), 0.98-1.02 (t, 3H).
Example 12
1- (4-Amino-1- (4-pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazol [3,4-d] pyrimidin-6-yl) pentan-1-one

Step 1
4 - ((4-Methoxybenzyl) amino) -1- (4- (pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazolo [3,4-d] pyrimidine-6-carbonitrile 12a [0208] Compound 1 (260 mg, 0.56 mmol), tris (dibenzylidenoacetone) dipaladium (52 mg, 0.056 mmol), 1,1'-bis (diphenylphosphino) ferrocene (31 mg, 0.056 mmol), zinc cyanide (99 mg, 0.84 mmol ) and zinc powder (37 mg, 0.56 mmol) were suspended in 5 ml of N, N-dimethylacetamide. The reaction solution was heated to 140 ° C and stirred for 16 hours under an argon atmosphere. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was
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82/97 purified by thin layer chromatography with development system A to obtain the title compound 12a (160 mg, yield: 63%).
[0209] MS m / z (ESI): 454.5 [M + l]
Step 2
1- (4 - ((4-Methoxybenzyl) amino) -1- (4- (pyrrolidin-1-ylmethyl 1) benzyl) -IH-pyrazolo [3,4-d] pyrimidin-6-11) pentan-l- one 12b [0210] Compound 12a (160 mg, 0.35 mmol) was dissolved in 5 mL of tetrahydrofuran and then a solution of 2 M n-butylmagnesium chloride in tetrahydrofuran (0.9 mL, 1.77 mmol) at 0 ° C. The reaction solution was heated to 60 ° C and stirred for 2 hours under an argon atmosphere. The reaction solution was cooled to room temperature, added with an aqueous solution of ammonium chloride and extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography with the development system to obtain the title compound 12b (150 mg, yield: 83%).
[0211] MS m / z (ESI): 513.6 [M + l]
Step 3
1- (4-Amino-1- (4-pyrrolidin-1-ylmethyl) benzyl) -IH-pyrazol [3,4-d] pyrimidin-6-11) pentan-1-one 12 [0212] Compound 12b (150 mg, 0.29 mmol) was dissolved in 10 ml of trifluoroacetic acid. The reaction solution was added to a sealed tube, heated to 110 ° C and stirred for
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83/97 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by high performance liquid chromatography (Waters-2767, elution system: 10 mmoL / L of ammonium bicarbonate, water, acetonitrile) to obtain the title compound 12 (19 mg, yield: 17%) .
[0213] MS m / z (ESI): 393.5 [M + l] [0214] ^X H NMR (400 MHz, CDC1 ₃ ): δ 8.01 (s, 1H), 7.35-7.29 (q, 4H), 5.62 (s, 2H), 3.62 (s, 2H), 3.26 (t, 2H), 2.53 (s, 4H), 1.78 (s, 4H) , 1.76-1.70 (m, 2H), 1.48-1.42 (m, 2H), 0.98 (t, 3H).
Test examples:
Biological Assay [0215] Test example 1. Determination of the agonistic activity of the compounds of the present invention in human TLR7 [0216] The activation effect of the compounds of the present invention on the hTLR7 protein expressed by the HET-Blue ™ hTLR7 stable transfected cells was determined by the following experimental method:
I. Materials and experimental instruments
1. DMEM (Gibco, 10564-029),
2. Fetal bovine serum (GIBCO, 10099),
3. Penicillin-streptomycin (Gibco, 15140-122),
4. Blue trypan solution (Sigma, T8154-100ML),
5. Flexstation 3 multifunction microplate reader
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84/97 (Molecular Devices),
6. HEK-Blue ™ HTLR7 cell line (InvivoGen, hkb-hTLR7),
7. HEK-Blue detection reagent (InvivoGen, hb-det3).
II. Experimental procedures [0217] A bag of dry HEK-Blue detection powder was dissolved in 50 ml of endotoxin-free water and the solution was then placed in an incubator at 37 ° C for 10 minutes followed by sterile filtration to prepare a HEK-Blue detection. The compound was initially formulated in a 20 mM stock solution, then diluted with pure DMSO to a maximum concentration of 6 x 10 ⁶ nM and a total of 10 points was obtained by a 3-fold gradient dilution.
[0218] The compound formulated above was first diluted 20 times with the medium, then 20 μΐ of the diluted compound was added to each well. The supernatant was removed from HEK-Blue ™ hTLR7 cells, to which 2-5 ml of pre-heated PBS was added. The cells were placed in an incubator for 1-2 minutes, pipetted gently and counted by trypan blue staining. The cells were resuspended in the HEK-Blue detection medium to adjust the concentration to 2.2 x 10 ⁵ cells / ml. 180 μΐ of cells was added to the 96-well plate above with 20 μΐ of the compounds and incubated at 37 ° for 6-16 hours.
[0219] The microplate reader read at a length of
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85/97 620 nm wave to obtain corresponding DO values, and the ECso values of the compounds were calculated by Graphpad Prism.
[0220] The activation effect of the compounds of the present invention on human TLR7 can be determined by the above test, and the ECso values obtained are shown in Table 1.
Table 1: ECso of the compounds of the present invention in human TLR7
No. Example ECso (nM) Emax (%) 1 28 100 2 64 91 3 77 91 4 166 88 6 233 91 7 180 95 8 217 104 9 128 96 10 349 79 11 335 85 12 388 78
[0221] Conclusion: The compounds of the present invention have a significant activation effect on human TLR7.
[0222] Test example 2. Determination of the agonistic activity of the compounds of the present invention in human TLR8 [0223] The activation effect of the compounds of the present invention on the hTLR8 protein expressed by the HET-Blue ™ hTLR8 stably transfected cells was determined by following experimental method:
I. Materials and experimental instruments
1. DMEM (Gibco, 10564-029),
2. Fetal bovine serum (GIBCO, 10099),
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3. Penicillin-streptomycin (Gibco, 15140-122),
4. Blue trypan solution (Sigma, T8154-100ML),
5. Flexstation 3 multifunction microplate reader (Molecular Devices),
6. HEK-Blue ™ HTLR8 cell line (InvivoGen, hkb-hTLR8),
7. HEK-Blue detection reagent (InvivoGen, hb-det3).
II. Experimental procedures [0224] A HEK-Blue detection dry powder bag was dissolved in 50 ml of endotoxin-free water and the solution was then placed in an incubator at 37 ° C for 10 minutes followed by sterile filtration to prepare a HEK-Blue detection. The compound was initially formulated in a 20 mM stock solution, then diluted with pure DMSO to a maximum concentration of 6 x 10 ⁶ nM and a total of 10 points was obtained by a 3-fold gradient dilution. The compound was first diluted 20 times with the medium, then 20 μΐ of the diluted compound was added to each well.
[0225] The supernatant was removed from HEK-Blue ™ hTLR8 cells, to which 2-5 ml of pre-heated PBS was added. The cells were placed in an incubator for 1-2 minutes, pipetted gently and counted by trypan blue staining. The cells were resuspended in the HEK-Blue detection medium to adjust the concentration to 2.2 x 10 ⁵ cells / ml. 180 μΐ of cells was added to the above 96-well plate with
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87/97 μΐ of the compounds and incubated at 37 ° for 6-16 hours.
[0226] The microplate reader read at a wavelength of 620 nm to obtain corresponding OD values, and the ECso values of the compounds were calculated by Graphpad Prism.
[0227] The activation effect of the compounds of the present invention on human TLR8 can be determined by the above test, and the ECso values obtained are shown in Table 2.
Table 2: ECso of the compounds of the present invention in human TLR8
No. Example ECso (μΜ) Emax (%) 1 > 30 8 2 > 29 52 3 > 24 2 4 > 30 28 6 > 6 35 7 > 30 0 8 > 30 2 10 > 30 0 11 > 30 0 12 > 30 5
[0228] Conclusion: The compounds of the present invention have no activation effect on human TLR8, indicating that the compounds of the present invention have a high selectivity on TLR7.
[0229] Test example 3. Determination of the ability of the compounds of the present invention to stimulate IFN-a secretion from peripheral blood mononuclear cells (PBMC) [0230] The ability of the compounds of the present invention to stimulate the secretion of IFN-α from PBMCs was determined by the following experimental method:
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I. Materials and experimental instruments
1. RPMI 1640 (Invitrogen, 11875),
2. FBS (Gibco, 10099-141)
3. Penicillin-streptomycin (Gibco, 15140-122),
4. Ficoll-Paque PREMIUM (GE, 17-5442-02),
5. Blue trypan solution (Sigma, T8154-100ML),
6. SepMateTM-50 (Stemcell, 15460),
7. Bright-Line ™ blood cell counter (Sigma, Z359629-1EA),
8. Human IFN-α kit (cisbio, 6FHIFPEB),
9. PHERAStar multifunction microplate reader (BMG, PHERAStar).
II. Experimental procedures [0231] The compound was diluted with pure DMSO to a minimum concentration of 5 mM and a total of 9 points was obtained by a 4-fold gradient dilution. 4 μΐ of the compound was then added to 196 μΐ of RMPI 1640 medium containing 10% FBS and mixed well. 50 μΐ of the mixture was removed from each well and added to a new 96-well plate.
[0232] All reagents were equilibrated at room temperature. 60 ml of blood and PBS + 2% FBS were added to a 250 ml culture flask, pipetted gently, mixed well and diluted. 15 ml of Ficoll-Paque PREMIUM lymphocyte separation solution and then 30 ml of diluted blood were added to 50 ml of centrifuge tube
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89/97 of SepMateTM-50 PBMC. The mixture was centrifuged at 1200 g for 10 minutes at room temperature. The supernatant was removed and then centrifuged at 300 g for 8 minutes. The cells were resuspended in RPMI 1640 medium containing 10% FBS and counted, and the number of PBMCs was adjusted to 3.33 x 10 ⁶ cells / ml. 150 μΐ of the cell solution was added to the plate added with the compounds, and incubated in an incubator at 37 ° C, in 5.0% CO2 for 24 hours.
[0233] The cell culture plate was placed in a centrifuge and centrifuged at 1200 rpm for 10 minutes at room temperature. 150 μΐ of the supernatant was taken from each well. The reagents in the human IFN-α kit were first equilibrated at normal temperature. The anti-IFN-a-Eu ³⁺ -Criptate conjugate and the anti-IFN-a-d2 conjugate were formulated in the dark according to the kit instructions, and both were well mixed with the conjugated buffer at a ratio of 1: 40. 16 μΐ of the supernatant obtained by centrifugation was then added to each well. 2 μΐ of anti-IFN-a-Eu ³⁺ -Cryptate conjugate and formulated anti-IFN-a-d2 conjugate are now added to each well. The plate was shaken and mixed well and incubated in the dark at room temperature for 3 hours.
[0234] PHERAStar reads in HTRF mode. The lowest concentration of compound that stimulates the cytokine level of at least 3 times more than the lower limit of detection was defined as
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90/97 the minimum effective concentration value (MEC) of the compound in the cytokine stimulation test.
[02 35] The ability of the compounds of the present invention to stimulate IFN-α secretion from PBMCs was determined by the above test, and the MEC values obtained are shown in Table
3.
Table 3: MEC of the compounds of the present invention to stimulate IFN-α secretion from PBMC
No. Example MEC (nM) 1 6 2 23 3 20 4 100 5 41 7 89
[0236] Conclusion: It can be seen from the activity data of stimulating IFN-α secretion from PBMCs that the compounds of the present invention have an advantage of less effective concentration.
[02 37] Test example 4. Effect of inhibiting the compounds of the present invention on the enzymatic activity of the CYP3A4 midazolam metabolite site in human liver microsomes [0238] The effect of the compounds of the present invention on the enzyme activity of the metabolite site of the midazolam of CYP3A4 in human liver microsomes was determined by the following experimental method:
I. Materials and experimental instruments
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1. Phosphate buffer (PBS),
2. NADPH (Sigma N-1630),
3. Human liver microsome (Corning Gentest),
4. ABI QTrap 4000 liquid chromatograph / mass spectrometer (AB Sciex),
5. InertsilC8-3 column, 4.6x50 mm, 5 pm (Dikma Technologies Inc., USA),
6. CYP probe substrate (midazolam / 10 μΜ) and positive control inhibitor (ketoconazole).
II. Experimental procedures [0239] The 100 mM PBS buffer was formulated, which was then used to formulate 2.5 mg / mL of microsome solution and 5 mM NADPH solution. The 5X concentration of the compound working solution was diluted with a PBS gradient (150, 50, 15, 5,
1.5, 0.15, 0.015, 0 μΜ). The 5X concentration of the ketoconazole working solution was diluted with a PBS gradient (150, 50, 15, 5, 1.5, 0.15, 0.015, 0 μΜ). The dextromethorphan working solution was diluted with PBS to a concentration of 50 μΜ.
[0240] 20 μΐ of 2.5 mg / ml microsome solution, 20 μΐ of 50 μΜ testosterone working solution, 20 μΐ of MgCÍ2 solution and 20 μΐ of the compound working solution (150, 50, 15, 5, 1.5, 0.15, 0.015, 0 μΜ, different reaction systems for each concentration) were taken, respectively, and well mixed. For the positive control group, the compound was
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92/97 replaced by the same concentration of ketoconazole. The mixture together with 5 mM NADPH solution was preincubated at 37 ° C for 5 minutes. After 5 minutes, 20 μΐ of NADPH were added to each well, the reaction was started and incubated for 30 minutes. All incubated samples were present in duplicate. After 30 minutes, 250 mL of acetonitrile containing internal standard were added to all samples, well mixed, stirred at 800 rpm for 10 minutes, and then centrifuged at 3700 rpm for 10 minutes. 80 μΐ of the supernatant were collected and analyzed by LC-MS / MS.
[0241] Data were calculated by Graphpad Prism to obtain the IC 50 values of the compounds at the CYP3A4 midazolam metabolite site.
[02 42] The ICso values of the compounds of the present invention at the CYP3A4 midazolam metabolite site in human liver microsomes.
No. Example ICso (μΜ) 1 14 2 10 3 7 4 11 6 10 7 > 30 12 16
[0243] Conclusion: The compounds of the present invention have
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93/97 a weak inhibitory effect on the metabolic site of CYP3A4 midazolam in the human liver microsome, and show better safety, indicating that the interaction of the metabolic drug based on the metabolic site of CYP3A4 midazolam will not occur.
[0244] Test example 5. Effect of inhibiting the compounds of the present invention on the enzymatic activity of CYP2D6 in human liver microsomes [0245] The effect of the compounds of the present invention on the enzymatic activity of CYP2D6 in human liver microsomes was determined by the following method experimental:
I. Materials and experimental instruments
1. Phosphate buffer (PBS),
2. NADPH (Sigma N-1630),
3. Human liver microsome (Corning Gentest),
4. ABI QTrap 4000 liquid chromatograph / mass spectrometer (AB Sciex),
5. InertsilC8-3 column, 4.6x50 mm, 5 pm (Dikma Technologies Inc., USA),
6. CYP probe substrate (dextromethorphan / 10 μΜ) and positive control inhibitor (quinidine).
II. Experimental procedures [0246] The 100 mM PBS buffer was formulated, which was then used to formulate 2.5 mg / ml of microsome solution and 5 mM NADPH solution. The 5X concentration of the compound working solution was diluted with a PBS gradient (150, 50, 15, 5,
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1.5, 0.15, 0.015, 0 μΜ). The 5Χ concentration of the compound's working solution was diluted with a PBS gradient (150, 50, 15, 5,
1.5, 0, 15, 0, 015, 0 μΜ). The dextromethorphan working solution was diluted with PBS to a concentration of 50 μΜ.
[0247] 20 μΐ of 2.5 mg / ml microsome solution, 20 μΐ of 50 μΜ testosterone working solution, 20 μΐ of MgCÍ2 solution and 20 μΐ of compound working solution (150, 50, 15, 5, 1.5, 0.15, 0.015, 0 μΜ, different reaction systems for each concentration) were taken, respectively, and well mixed. For the positive control group, the compound was replaced by the same concentration of quinidine. The mixture together with 5 mM NADPH solution was preincubated at 37 ° C for 5 minutes. After 5 minutes, 20 μΐ of NADPH was added to each well, the reaction was started and incubated for 30 minutes. All incubated samples were present in duplicate. After 30 minutes, 250 mL of acetonitrile containing internal standard were added to all samples, well mixed, stirred at 800 rpm for 10 minutes, and then centrifuged at 3700 rpm for 10 minutes. 80 μΐ of the supernatant were collected and analyzed by LC-MS / MS.
[0248] Data were calculated by Graphpad Prism to obtain the ICso values of the compounds at the CYP2D6 metabolite site.
[024 9] The ICso values of the compounds of the present invention by no inhibition of CYP2D6 in human liver microsomes.
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No. Example ICso (μΜ) 1 > 30 2 > 30 3 4 4 16 6 10 7 > 30 12 16
[0250] Conclusion: The compounds of the present invention have a weak inhibiting effect on the enzymatic activity of CYP2D6 in human liver microsomes, and show better safety, indicating that the metabolic interaction based on CYP2D6 will not occur.
[0251] Test example 6. Inhibiting effect of the compounds of the present invention on the enzymatic activity of the CYP3A4 testosterone metabolite site in human liver microsomes.
[0252] The effect of the compounds of the present invention on the enzymatic activity of the CYP3A4 testosterone metabolite site in human liver microsomes was determined by the following experimental method:
I. Materials and experimental instruments
1. Phosphate buffer (PBS),
2. NADPH (Sigma N-1630),
3. Human liver microsome (Corning Gentest),
Petition 870190046717, of 5/20/2019, p. 111/130
96/97
4. ABI QTrap 4000 liquid chromatograph / mass spectrometer (AB Sciex),
5. InertsilC8-3 column, 4.6x50 mm, 5 pm (Dikma Technologies Inc., USA),
6. CYP probe substrate (testosterone / 10 μΜ) and positive control inhibitor (ketoconazole).
II. Experimental procedures [0253] The 100 mM PBS buffer was formulated, which was then used to formulate 2.5 mg / ml microsome solution and 5 mM NADPH solution. The 5X concentration of the compound working solution was diluted with a PBS gradient (150, 50, 15, 5,
1.5, 0.15, 0.015, 0 μΜ). The 5X concentration of the ketoconazole working solution was diluted with a PBS gradient (150, 50, 15, 5, 1.5, 0.15, 0.015, 0 μΜ). The dextromethorphan working solution was diluted with PBS to a concentration of 50 μΜ.
[0254] 20 μΐ of 2.5 mg / ml microsome solution, 20 μΐ of 50 μΜ testosterone working solution, 20 μΐ of MgC12 solution and 20 μΐ of compound working solution (150, 50, 15, 5, 1.5, 0.15, 0.015, 0 μΜ, different reaction systems for each concentration) were taken, respectively, and well mixed. For the positive control group, the compound was replaced by the same concentration of ketoconazole. The mixture together with 5 mM NADPH solution was preincubated at 37 ° C for 5 minutes. After 5 minutes, 20 μΐ of NADPH were added
Petition 870190046717, of 5/20/2019, p. 112/130
97/97 at each well, the reaction was started and incubated for 30 minutes. All incubated samples were present in duplicate. After 30 minutes, 250 mL of acetonitrile containing internal standard were added to all samples, well mixed, stirred at 800 rpm for 10 minutes, and then centrifuged at 3700 rpm for 10 minutes. 80 μΐ of the supernatant were collected and analyzed by LC-MS / MS.
[0255] Data were calculated by Graphpad Prism to obtain the IC 50 values of the compounds at the CYP3A4 testosterone metabolite site.
[0256] The IC 50 values of the compounds of the present invention at the CYP3A4 testosterone metabolite site in human liver microsomes.
No. Example ICso (μΜ) 1 4 2 19 3 3 4 6 6 3 7 > 30 12 > 30
[0257] Conclusion: The compounds of the present invention have a weak inhibition in the metabolite site of CYP3A4 testosterone in human liver microsomes, and show better safety.

权利要求:
Claims (24)
[1]
1. Compound of formula (I):

[2]
2/14 alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ^S R ⁷ and -C (O) NR ⁶ R ⁷ ;
each R ² is the same or different and each is independently selected from the group consisting of hydrogen, halogen, alkyl, alkoxy, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl, where alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl , aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ^S R ⁷ and -C (O) NR ⁶ R ⁷ ;
L ² is alkylene or a covalent bond, where the alkylene is optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl , aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ^S R ⁷ and -C (O) NR ⁶ R ⁷ ;
R ³ is selected from the group consisting of haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ ,
Petition 870190046717, of 5/20/2019, p. 115/130
[3]
3/14
-S (O) _m R ⁵ , -NR ⁶ R ⁷ eC (0) NR ⁶ R ⁷ , where cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted by one or more substituents selected from the group that consists of alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁸ , -C (O) R ⁸ , -S (O) _m R ⁸ , - NR ⁹ R ¹⁰ and -C (O) NR ⁹ R ¹⁰ ;
R ⁴ is selected from the group consisting of hydrogen, alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;
R ⁵ is selected from the group consisting of hydrogen, alkyl, haloalkyl, amino, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl;
R ⁶ and R ⁷ are identical or different and are each independently selected from the group consisting of hydrogen, alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C (O) R ⁸ , -S (O) _m R ⁸ and -C (O) NR ⁹ R ¹⁰ , where the alkyl, cycloalkyl, heterocyclyl, arylheteroaryl are each independently optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, amino , cyano, nitro, hydroxy hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;
or, R ⁶ and R ⁷ together with the nitrogen to which they are attached, form a heterocyclyl, where the heterocyclyl optionally contains one or two identical or different heteroatoms
Petition 870190046717, of 5/20/2019, p. 116/130
[4]
4/14 selected from the group consisting of N, 0 and S, in addition to a nitrogen atom, and the heterocyclyl is optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;
R ⁸ is selected from the group consisting of hydrogen, alkyl, haloalkyl, amino, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl;
R ⁹ and R ¹⁰ are identical or different and are each independently selected from the group consisting of hydrogen, alkyl, haloalkyl, amino, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl;
neO, 1, 2, 3 or 4; and m is 0, 1 or 2.
2. A compound of formula (I) according to claim 1, characterized by the fact that R ³ is heterocyclyl, and the heterocyclyl is optionally substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, halogen , amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl.
3. A compound of formula (I), according to claim 1 or 2, characterized by the fact that R ³ is -NR ⁶ R ⁷ , and R ⁶ and R ⁷ together with the nitrogen to which they are attached, form a heterocyclyl, where the heterocyclyl optionally contains a
Petition 870190046717, of 5/20/2019, p. 117/130
[5]
5/14 or two identical or different heteroatoms selected from the group consisting of N, 0 and S in addition to a nitrogen atom and the heterocyclyl are optionally substituted with one or more substituents selected from the group consisting of alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl.
4. Compound of formula (I) according to claim
1 or 2, characterized by the fact that ring A is phenyl.
Compound of formula (I) according to any one of claims 1 to 4, characterized in that X ¹ is alkylene.
[6]
A compound of formula (I) according to any one of claims 1 to 5, characterized in that it is a compound of formula (II):
NH.

[7]
Compound of formula (I) according to any one of claims 1 to 6, characterized by the fact that G is N.
[8]
Compound of formula (I) according to any one of claims 1 to 7, characterized in that L ² is alkylene.
[9]
Compound of formula (I) according to any one of claims 1 to 8, characterized in that it is a compound of formula (III):
NH ₂

[10]
10. Compound of formula (I) according to any one of claims 1 to 9, characterized by the fact that L ¹ is selected from the group consisting of -O-, -NR ⁴ -, -C (O) - and -C (O) N (R ⁴ ) -, and R ⁴ is hydrogen or alkyl.
[11]
A compound of formula (I) according to any one of claims 1 to 10, characterized in that R ¹ is alkyl optionally substituted by one or more alkoxy groups.
Petition 870190046717, of 5/20/2019, p. 119/130
7/14
[12]
A compound of formula (I) according to any one of claims 1 to 11, characterized in that each R ² is identical or different and each is independently hydrogen or halogen.
[13]
13. A compound of formula (I) according to any one of claims 1 to 12, characterized by the fact that it is selected from the group consisting of:

[14]
14. Compound of formula (I-C):

[15]
15. Compound of formula (I-C), according to claim 14, characterized by the fact that it is selected from the group that
Petition 870190046717, of 5/20/2019, p. 121/130
9/14 consists of:

[16]
16. Compound of formula (I-E):
HN — W

[17]
17. Compound of formula (I-E) according to claim
16, characterized by the fact that it is selected from the group consisting of:

[18]
18. Method for preparing the compound of formula (I-E) according to claim 16, characterized in that it comprises a step of:

[19]
19. Method for preparing the compound of formula (I) according to claim 1, characterized in that it comprises a step of:
NH.
removing the protecting group from a compound of formula (I-E) under an acidic condition to obtain the compound of formula (I); on what:
W is an amino protecting group;
ring A, G, VL ² , X ¹ , R ² ~ R ^{3 and} n are as defined in claim 1.
[20]
20. Pharmaceutical composition, characterized by the fact that it comprises a therapeutically effective amount of the compound, or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or a mixture thereof, or a salt
Petition 870190046717, of 5/20/2019, p. 125/130
Pharmaceutically acceptable 13/14 thereof as defined in any one of claims 1 to 13, and one or more pharmaceutically acceptable carriers, diluents or excipients.
[21]
21. Use of the compound, or a tautomer, mesomer, racemate, enantiomer, diastereoisomers thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof, as defined in any of claims 1 to 13, or the pharmaceutical composition, as defined in claim 20 characterized by the fact that it is in the preparation of a drug to activate TLR7.
[22]
22. Use of the compound, or a tautomer, mesomer, racemate, enantiomer, diastereoisomers thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof, as defined in any of claims 1 to 13, or the pharmaceutical composition, as defined in claim 20, characterized by the fact that it is the preparation of a medicine for the treatment of an infection caused by a virus selected from the group consisting of dengue virus, yellow fever virus, West Nile virus, Japanese encephalitis virus, tick-borne encephalitis, Kunjin virus, Murray Valley encephalitis virus, St. Louis encephalitis virus, Omsk hemorrhagic fever virus, bovine virus disease virus, Zika virus, HIV virus, HBV, HCV, HPV, RSV , SARS and influenza.
[23]
23. Use of the compound, or a tautomer, mesomer,
Petition 870190046717, of 5/20/2019, p. 126/130
Racemate, enantiomer, diastereoisomers thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, as defined in any of claims 1 to 13, or the pharmaceutical composition, as defined in claim 20, characterized by the fact that be in the preparation of a medication for the treatment or prevention of melanoma, non-small cell lung carcinoma, hepatocellular carcinoma, basal cell carcinoma, renal cell carcinoma, myeloma, allergic rhinitis, asthma, COPD, ulcerative colitis and liver fibrosis.
[24]
24. A compound, or a tautomer, mesomer, racemate, enantiomer, diastereoisomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof, as described in any one of claims 1 to 13, or a pharmaceutical composition comprising the same, characterized by the fact that it is for use as a medicine.

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法律状态:
2021-10-05| B06W| Patent application suspended after preliminary examination (for patents with searches from other patent authorities) chapter 6.23 patent gazette]|

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2021-10-05| B350| Update of information on the portal [chapter 15.35 patent gazette]|

优先权:

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申请号 | 申请日 | 专利标题

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CN201611066071|2016-11-28|

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PCT/CN2017/113007|WO2018095426A1|2016-11-28|2017-11-27|Pyrazolo-heteroaryl derivative, preparation method and medical use thereof|

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