CN109134463B

CN109134463B - β -carboline type 5 phosphodiesterase inhibitor and preparation method and application thereof

Info

Publication number: CN109134463B
Application number: CN201811081111.4A
Authority: CN
Inventors: 娄红祥; 郑泓波; 孙斌
Original assignee: Shandong University
Current assignee: Shandong University
Priority date: 2018-09-17
Filing date: 2018-09-17
Publication date: 2020-05-08
Anticipated expiration: 2038-09-17
Also published as: CN109134463A

Abstract

The invention provides an β -carboline type 5 phosphodiesterase inhibitor, a preparation method and application thereof, wherein the inhibitor has a structure shown in a formula (X):

wherein X is selected from sulfur and oxygen; r₁Is a substituent on the five-membered heterocyclic ring, R₁The substituent can be one or more, and each substituent is independently selected from hydrogen, bromine and C₁‑C₄Alkyl and phenyl groups of (a), including substituted and unsubstituted phenyl groups; the substituted or unsubstituted phenyl and the five-membered heterocyclic ring in which the substituted or unsubstituted phenyl is located form a condensed ring by sharing a ring edge; the A group is fused to the five-membered ring of the indole with a common ring edge, the A group being selected from:

R₂is selected from C₂‑C₄Alkyl groups of (a); r₃Selected from bipyridyl, 4- (3, 4-dimethoxyphenyl) pyrimidinyl; or R₃and-NH-ring to form the structure:

R₄selected from the group consisting of bromine, carbomethoxy and carboxyl; and, the compounds of formula (X) also include optically pure enantiomers thereof.

Description

β -carboline type 5 phosphodiesterase inhibitor and preparation method and application thereof

Technical Field

The invention relates to the field of medicines, and particularly relates to an β -carboline type 5 phosphodiesterase inhibitor, and a preparation method and application thereof.

Background

Phosphodiesterases (PDEs) are a large family of polygenes, comprising more than 30 distinct phosphodiesterase isoenzymes of type 11. Phosphodiesterases (PDEs) have the function of hydrolyzing intracellular second messengers (cAMP, cyclic adenosine monophosphate or cGMP, cyclic guanosine monophosphate) and thus regulate a variety of important physiological processes. The development of selective phosphodiesterase inhibitors will open new avenues for the treatment of a variety of diseases.

Phosphodiesterase type 5 (PDE5) is capable of specifically hydrolyzing cGMP and regulating the levels of intracellular cGMP, and is involved in a variety of important biophysiological processes in vivo. PDE5 is widely distributed in the human lung, in the corpus cavernosum smooth muscle of the penis, and in vascular, visceral and airway smooth muscle, skeletal muscle, platelets and other tissues. Selective PDE5 inhibitors act primarily on the Nitric Oxide (NO) -soluble guanylyl cyclase (sGC) -cyclic guanosine monophosphate (cGMP) pathway and are able to maintain intracellular high concentrations of cGMP by inhibiting the hydrolysis process of cGMP by PDE 5. Therefore, PDE5 is an ideal drug target for treating diseases related to cGMP level, such as Male Erectile Dysfunction (MED), Pulmonary Arterial Hypertension (PAH), etc. In addition, long-term clinical treatment procedures found that various PDE5 inhibitors had significant improvement effects on cognitive function in patients with alzheimer's disease, which may be correlated with the effects of PDE5 inhibitors on vasodilation and improvement of blood flow (j.med.chem.doi: 10.1021/acs.jmedchem.7b01370).

Three PDE5 inhibitor medicines sold in China

Currently, commercially available drugs of PDE5 inhibitors sold in China mainly comprise nucleoside derivatives Sildenafil (Sildenafil), Vardenafil (Vardenafil) and β -carboline compounds Tadalafil (Tadalafil), wherein the Tadalafil has a unique β -carboline skeleton, and a plurality of reports show that β -carboline compounds have remarkable PDE5 inhibition activity, such as the following compounds 1-4.

The inventor of the application synthesizes and reports β -carboline alkaloid perolyrin in caltrops and analogues 5a-b thereof in advance to have weak PDE5 inhibition activity on PDE5, and carries out further structure optimization and synthesizes a series of furyl or thienyl derivatives on the basis of the weak PDE5 inhibition activity, and the results show that the derivatives containing 5-ethylfuran or 5-ethylfuran thiophene, such as the following compounds 6-7, have very remarkable PDE5 inhibition activity, however, the compounds have poor selectivity on PDE6 and PDE11 and are worthy of further structure optimization and activity screening.

The crystal complex of tadalafil and PDE5 protein shows that β -carboline derivatives can be well combined with the active pocket of PDE5 and show a unique combination mode to further exert good PDE5 inhibition activity (Nature,2003,425, 99-102). the NH structure of indole part in tadalafil structure can form hydrogen bond action with the side chain of Gln 817 in protein, and the C-6 substituted aryl part can be combined with Q of PDE5₂The β -carboline compounds are structurally optimized based on the combination mode of ligand and target, which is beneficial to developing new β -carboline PDE5 inhibitors with high activity and high selectivity.

Disclosure of Invention

Therefore, the invention aims to provide a novel β -carboline PDE5 inhibitor with high activity and high selectivity, and a preparation method and application thereof.

The application is realized by the following technical scheme:

first, the present application provides a class of compounds, or pharmaceutically acceptable salts thereof, having a structure as shown in formula (X):

wherein X is selected from sulfur and oxygen;

R₁is a substituent on the five-membered heterocyclic ring, R₁The substituent can be one or more, and each substituent is independently selected from hydrogen, bromine and C₁-C₄Alkyl and phenyl groups of (a), including substituted and unsubstituted phenyl groups; the substituted or unsubstituted phenyl and the five-membered heterocyclic ring in which the substituted or unsubstituted phenyl is located form a condensed ring by sharing a ring edge; the A group being common to the five-membered ring of the indoleThe ring sides are fused, and the A group is selected from:

R₂is selected from C₂-C₄Alkyl groups of (a); r₃Selected from bipyridyl, 4- (3, 4-dimethoxyphenyl) pyrimidinyl; or R₃with-NH-ring to form a radical selected from

A group of (a); r₄Selected from the group consisting of bromine, carbomethoxy and carboxyl; and, the compounds of formula (X) also include optically pure enantiomers thereof.

The above A group or R₃The groups formed by ring closure with-NH-contain 1 or 2 asymmetric chiral carbon atoms, respectively, and exist in diastereoisomeric and enantiomeric forms.

Preferably, the first and second electrodes are formed of a metal,

is composed of

Preferably, the first and second electrodes are formed of a metal,

is composed of

Preferably, the first and second electrodes are formed of a metal,

is composed of

Preferably, the first and second electrodes are formed of a metal,

is composed of

Preferably, the first and second electrodes are formed of a metal,

is composed of

Preferably, the compound of formula (X) has the structure of formula (I) -formula (VII):

wherein the content of the first and second substances,

represents a single bond or a double bond. When in use

When the double bond is formed, the compound of the formula (II) is a double bond product after dehydrogenation of C-11 and C-11a position, and the structure of the compound of the formula (II) is

Preferably, R is in the structure of formula (I) to formula (VII) above₁The number of the substituent groups is 1 or 2, and the substitution position is selected from C-3, C-4 and C-5; preferably, R₁Selected from hydrogen, bromine, C₁-C₃Alkyl and phenyl groups of (a), which phenyl groups may be substituted or unsubstituted; wherein when R is₁In the case of a phenyl group, the substituted or unsubstituted phenyl group may form a condensed ring with a common ring with the five-membered heterocyclic ring in which it is present; for example, when X is oxygen and R1 is phenyl, the phenyl group may form a phenylpropyl furyl group with a common side to the furyl group.

Preferably, R is in the structure of formula (I) to formula (VII) above₂Selected from ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl; preferably, R is in the structure of formula (I) to formula (VII) above₃Selected from bipyridyl and 4- (3, 4-dimethoxyphenyl) pyrimidinyl;

or at

In, R₃with-NH-ring to form a radical selected from

A group of (a);

as mentioned above, the compounds of the present invention also comprise optically pure enantiomers thereof, which contain 1 or 2 asymmetric chiral carbon atoms, as compared to the compounds of formula (I) -formula (III) and formula (V) -formula (VII), respectively, and exist in the form of diastereomers and enantiomers.

Preferably, the chiral centers of the compounds of formula (I) are the C-6 and C-12a positions; the chiral center of the formula (II) is C-5 and C-11a site; the chiral center of the formula (III), the formula (V) -the formula (VI) is C-1 position; the chirality of the compound of formula (IV) is C-3; the chiral centers of the compound of formula (VII) are the C-1 and C-3 positions.

Preferably, in the compounds of formula (I) according to the invention, X is selected from O and S; r ₁1 substituent selected from hydrogen, bromine, methyl and ethyl; r₁The substitution position(s) is selected from the C-3 and C-5 positions, preferably the C-5 position; the compounds of formula (I) have two chiral centres, respectively at the C-6 and C-12a positions, and have four isomers selected from (6S,12aS), (6R,12aR), (6R,12aS) and (6S,12aR), with the compounds of formula (I) being in any of the four configurations, preferably (6S,12aR) or (6R,12 aS).

Preferably, in the compounds of formula (II) of the present invention, X is selected from O and S; r ₁1 substituent selected from methyl and ethyl, R₁The substitution position(s) is selected from the C-3 and C-5 positions, preferably the C-5 position; r₂Is selected from C₂-C₄Alkyl groups of (a); in the compound of formula (II)

Is a single bond or a double bond. In the compound of formula (II)

When the compound is a double bond, the compound of the formula (II) is a C-11, C-11a dehydrogenated bisA bond product of i

Wherein, in the compound of formula (II), R₂Selected from ethyl, propyl, isopropyl, n-butyl, isobutyl and tert-butyl, preferably from ethyl and n-butyl; the two chiral centers of formula (II), which are C-5 and C-11a, have four isomers selected from (5S,11aS), (5R,11aR), (5R,11aS) and (5S,11aR), and formula (II) can have any one of four configurations, preferably (5S,11aS) or (5S,11 aR).

Preferably, in the compounds of formula (III) according to the invention, X is selected from O and S; r₁The substituent is 1 or 2 and is selected from C₁-C₃And phenyl, which may be substituted or unsubstituted, preferably said substituted phenyl is phenyl substituted with alkoxy; r₃Selected from bipyridyl and 4- (3, 4-dimethoxyphenyl) pyrimidinyl, preferably 4- (3, 4-dimethoxyphenyl) pyrimidinyl; preferably, R₁The substitution position(s) is selected from the C-3, C-4 and C-5 positions; preferably, R₁Selected from methyl, ethyl and n-propyl.

Preferably, in the compound of formula (III), X is O or S; r₁The substituent is 1, R₁When it is phenyl or substituted phenyl, R₁Form a condensed ring with the five-membered heterocyclic ring in which the five-membered heterocyclic ring is positioned by sharing a ring edge; wherein, the substituted phenyl is phenyl substituted by alkoxy, preferably methoxyphenyl, and the substituted position of the alkoxy on the phenyl is preferably C-7 position.

Preferably, in the compound of formula (III), X is O or S; r₁When the substituent is 1, R₁Is methyl, ethyl or n-propyl, R₁The substitution position of (A) is C-5 or C-3.

Preferably, in the compound of formula (III), X is O or S; r₁When the number of the substituents is 2, the substituents are preferably all methyl groups, and the substitution positions are preferably the C-4 position and the C-5 position, respectively.

Preferably, in the compound of formula (III), X is O, R₁When it is phenyl or substituted phenyl, R₁Condensed with furyl radicals in common ring-sides, substituted or unsubstitutedA benzofuranyl group; the substituted phenyl group is a phenyl group substituted with an alkoxy group, preferably a methoxyphenyl group.

Preferably, the chiral centre of the compound of formula (III) is the C-1 position.

Preferably, in the compounds of formula (IV) of the present invention, preferably, in the compounds of formula (IV), X is selected from O and S;

r

₁1 or 2 substituents R₁The substituents being selected from C₁-C₃Alkyl and phenyl of (a), which phenyl may be substituted or unsubstituted phenyl; r₃Selected from bipyridyl, 4- (3, 4-dimethoxyphenyl) pyrimidinyl, or R₃with-NH-ring to form a radical selected from

A group of (1).

Preferably, in the compound of formula (IV), R₁The substitution position(s) is selected from the C-3, C-4 and C-5 positions.

Preferably, in the compound of formula (IV), X is O or S; r₁The substituent is 1, R₁When it is phenyl or substituted phenyl, R₁Form a condensed ring with the five-membered heterocyclic ring in which the five-membered heterocyclic ring is positioned by sharing a ring edge; wherein the substituted phenyl is phenyl substituted by alkoxy, preferably methoxyphenyl; the substitution position of the alkoxy group on the phenyl group is preferably the C-7 position.

Preferably, in the compound of formula (IV), X is O or S; r₁The substituent is 1, R₁Is methyl, ethyl or n-propyl, R₁The substitution position of (A) is C-5 or C-3.

Preferably, in the compound of formula (IV), X is O or S; r₁When the number of the substituents is 2, the substituents are preferably all methyl groups, and the substitution positions are preferably the C-4 position and the C-5 position, respectively.

Preferably, in the compound of formula (IV), X is O, R₁When it is phenyl or substituted phenyl, R₁Condensed with furyl in a common ring edge to form substituted or unsubstituted benzofuryl; the substituted phenyl is phenyl substituted by alkoxy, preferably methoxyphenyl; the substitution position of the alkoxy group on the phenyl group is preferably the C-7 position.

Preferably, a compound of formula (IV)In which X is S, R₁The substituent is 1, R₁The substituents being methyl, ethyl or n-propyl, R₁The substitution position of (A) is C-5 or C-3, preferably C-5, R₃By ring closure with-NH-groups

Preferably, the chiral center of the compound of formula (IV) is the C-3 position and the compound of formula (IV) is in the R or S configuration, preferably the S configuration.

Preferably, in the compound of formula (VII) of the present invention, the compound of formula (VII) has 2 chiral centers, respectively at C-1 and C-3 positions, which are any one of four isomers selected from (1R,3S), (1R,3R), (1S,3S) and (1S,3R), preferably in the (1R,3S) configuration.

Preferably, the above compound or an optically pure enantiomer or solvate or hydrate thereof or a pharmaceutically acceptable salt thereof is selected from the following compounds:

(6R,12aR) -6- (furan-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6S,12aR) -6- (furan-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6R,12aR) -2-methyl-6- (5-methylfuran-2-yl) -2,3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6S,12aR) -2-methyl-6- (5-methylfuran-2-yl) -2,3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6R,12aR) -2-methyl-6- (5-methylthiophen-2-yl) -2,3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6S,12aR) -2-methyl-6- (5-methylthiophen-2-yl) -2,3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6R,12aR) -2-methyl-6- (3-methylthiophen-2-yl) -2,3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6S,12aR) -2-methyl-6- (3-methylthiophen-2-yl) -2,3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6R,12aR) -6- (5-ethylfuran-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6S,12aR) -6- (5-ethylfuran-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6S,12aS) -6- (5-ethylfuran-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6R,12aS) -6- (5-ethylfuran-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6R,12aR) -6- (3-bromothiophen-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6S,12aR) -6- (3-bromothien-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6R,12aR) -6- (5-bromothiophen-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6S,12aR) -6- (5-bromothien-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6R,12aR) -6- (5-ethylthiophen-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6S,12aR) -6- (5-ethylthiophen-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6S,12aS) -6- (5-ethylthiophen-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(6R,12aS) -6- (5-ethylthiophen-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyridine [3,4-b ] indole-1, 4-dione;

(5S,11aS) -2-ethyl-5- (5-ethylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5R,11aS) -2-ethyl-5- (5-ethylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5R,11aR) -2-ethyl-5- (5-ethylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5S,11aR) -2-ethyl-5- (5-ethylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5S,11aS) -2-ethyl-5- (5-ethylfuran-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5R,11aR) -2-ethyl-5- (5-ethylfuran-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5S,11aR) -2-ethyl-5- (5-ethylfuran-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5S,11aS) -2-ethyl-5- (3-methylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5R,11aS) -2-ethyl-5- (3-methylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5S,11aS) -2-ethyl-5- (5-methylfuran-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5S,11aS) -2-butyl-5- (5-ethylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5R,11aS) -2-butyl-5- (5-ethylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5R,11aR) -2-butyl-5- (5-ethylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

(5S,11aR) -2-butyl-5- (5-ethylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6] pyridine [3,4-b ] indole-1, 3(2H) -dione;

n-ethyl-1- (5-ethylthiophen-2-yl) -9-hydro-pyrrolo [3,4-b ] indole-3-carboxamide;

2- ([2,3 '-bipyridin ] -6' -yl) -1- (4, 5-dimethylthiophen-2-yl) -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indole;

2- ([2,3 '-bipyridin ] -6' -yl) -1- (benzofuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indole;

2- ([2,3 '-bipyridin ] -6' -yl) -1- (7-methoxybenzofuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indole;

2- ([2,3 '-bipyridin ] -6' -yl) -1- (5-methylthiophen-2-yl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole;

2- ([2,3 '-bipyridin ] -6' -yl) -1- (5-propylthiophen-2-yl) -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indole;

2- ([2,3 '-bipyridin ] -6' -yl) -1- (5-methylfuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole;

2- (5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) -1- (5-ethylfuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indole;

2- (5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) -1- (5-ethylthiophen-2-yl) -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indole;

(3- (2- ([2,3 '-bipyridin ] -6' -ylamino) ethyl) -1H-indol-2-yl) (5-ethylthiophen-2-yl) methanone;

(3- (2- ([2,3 '-bipyridin ] -6' -ylamino) ethyl) -1H-indol-2-yl) (4, 5-dimethylthiophen-2-yl) methanone;

(3- (2- ([2,3 '-bipyridin ] -6' -ylamino) ethyl) -1H-indol-2-yl) (5-propylthiophen-2-yl) methanone;

(3- (2- ([2,3 '-bipyridin ] -6' -ylamino) ethyl) -1H-indol-2-yl) (5-methylthiophen-2-yl) methanone;

(3- (2- ([2,3 '-bipyridin ] -6' -ylamino) ethyl) -1H-indol-2-yl) (5-ethylfuran-2-yl) methanone;

(3- (2- ([2,3 '-bipyridin ] -6' -ylamino) ethyl) -1H-indol-2-yl) (5-methylfuran-2-yl) methanone;

(3- (2- ([2,3 '-bipyridin ] -6' -ylamino) ethyl) -1H-indol-2-yl) (benzofuran-2-yl) methanone;

(3- (2- ([2,3 '-bipyridin ] -6' -ylamino) ethyl) -1H-indol-2-yl) (7-methoxybenzofuran-2-yl) methanone;

(3- (2- ((5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) amino) ethyl) -1H-indol-2-yl) (5-ethylfuran-2-yl) methanone;

(3- (2- ((5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) amino) ethyl) -1H-indol-2-yl) (5-ethylthiophen-2-yl) methanone;

(S) -3- ((2- (5-ethylthiophene-2-carbonyl) -1H-indol-3-yl) methyl) -1-methylpiperazine-2, 5-dione;

(R) -3- ((2- (5-ethylthiophene-2-carbonyl) -1H-indol-3-yl) methyl) -1-methylpiperazine-2, 5-dione;

(S) -3-ethyl-5- ((2- (5-ethylthiophene-2-carbonyl) -1H-indol-3-yl) methyl) imidazoline-2, 4-dione;

(R) -3-ethyl-5- ((2- (5-ethylthiophene-2-carbonyl) -1H-indol-3-yl) methyl) imidazoline-2, 4-dione;

2- (5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) -1- (5-ethylfuran-2-yl) -1,2,3, 9-tetrahydro-4H-pyridin [3,4-b ] indol-4-one;

2- (5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) -1- (5-ethylthiophen-2-yl) -1,2,3, 9-tetrahydro-4H-pyridin [3,4-b ] indol-4-one;

methyl- (E) -4- (3- (1- (5-ethylthiophen-2-yl) -1,3,4, 9-tetrahydro-2H-pyridine [3,4-b ] indol-2-yl) -3-oxo-1-en-1-yl) benzoate;

methyl- (E) -4- (3- (1- (5-ethylthiophen-2-yl) -1,3,4, 9-tetrahydro-2H-pyridine [3,4-b ] indol-2-yl) -3-oxo-1-en-1-yl) benzoic acid;

(E) -3- (4-bromophenyl) -1- (1- (5-ethylthiophen-2-yl) -1,3,4, 9-tetrahydro-2H-pyridine [3,4-b ] indol-2-yl) propenyl-1-one;

methyl (1R,3S) -2- (ethylcarbamoyl) -1- (5-ethylfuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indole-3-carboxylate.

Methyl (1R,3S) -2- (ethylcarbamoyl) -1- (5-ethylthiophen-2-yl) -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indole-3-carboxylate.

Preferably, the present invention also includes pharmaceutically acceptable salts, solvates, hydrates, prodrug compounds, polymorphs and corresponding active metabolites of the above compounds; preferably, the pharmaceutically acceptable salts include salts of the above compounds with inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid or hydrobromic acid, and salts with organic acids such as methanesulfonic acid, toluenesulfonic acid or trifluoroacetic acid.

The compounds contain 1 or 2 asymmetric chiral carbon atoms compared with the compounds shown in formula (I) -formula (III) and formula (V) -formula (VII), and exist in the forms of diastereoisomers and enantiomers.

There is a significant difference in the inhibitory activity against phosphodiesterase type 5 between the different isomers. Therefore, the invention also comprises 2-4 diastereoisomers or enantiomers, racemes or mixtures thereof of the compounds shown in the formula (I) -formula (III) and the formula (V) -formula (VII) in any proportion. The preparation and separation of diastereoisomers can be carried out by conventional column chromatography and the like, and the separation of enantiomers can be carried out by conventional chiral resolution methods. For example, compounds of formula (III) may be resolved by chiral HPLC chromatography or by fractional crystallization of the diastereomeric salt formed by reaction of the racemate with an appropriate optically active acid or base; the compound of formula (III) can also be prepared by carrying out the corresponding chemical reaction on the optically pure compound of formula (I). Optically pure compounds of formula (I) -formula (III), formula (V) -formula (VII) may also be prepared by chemical reaction under suitable catalytic conditions.

The application also provides a method for preparing the compound or an optically pure enantiomer or a solvate or a hydrate or a pharmaceutically acceptable salt thereof, which comprises the step of carrying out cross dehydrogenation coupling reaction on β -carboline substrate and an organic boron reagent under an oxidation condition to generate α substituted β -carboline derivatives, namely the compound of formula (I) -formula (VII);

preferably, the β -carboline substrate is

Wherein the A group is fused to the five-membered ring of the indole in a common ring edge, the A group being selected from:

R₁is mono-or di-substituted, and the substituent is selected from hydrogen, bromine and C₁-C₃Including substituted and unsubstituted phenyl groups, including substitution of substituents on and with the ring in which they are located to form a fused ring with the common edge; r₂Is selected from C₁-C₄Alkyl groups of (a); r₃Selected from bipyridyl, 4- (3, 4-dimethoxyphenyl) pyrimidinyl, substituted pyrazine ring, and substituted imidazole ring; r₄Selected from the group consisting of bromine, carbomethoxy and carboxyl;

the bond is a single bond or a double bond.

Preferably, the β -carboline substrate is selected from compounds 12, 14, and 17 shown below:

wherein R is selected from

Preferably, the oxidizing agent in the reaction includes, but is not limited to, 2,6, 6-tetramethylpiperidine-nitrogen-oxide (TEMPO);

preferably, the organoboron reagent is of heterocyclic structure

Fluoroborate, borate, boric acid of (1), wherein R₁Is as defined above;

preferably, the process is carried out as follows:

preferably, the process is carried out as follows:

wherein, X, R₁Is as defined above;

alternatively, the process is carried out as follows:

wherein, X, R₁、R₂Is as defined above;

alternatively, the process is carried out as follows:

wherein, X, R₁、R₃Is as defined above;

alternatively, the process is carried out as follows:

wherein the content of the first and second substances,X、R₁、R₄is as defined above.

Preferably, the compound 12 is prepared by the following reaction:

(D) tryptophan or tryptophan hydrochloride ((D) -tryptophan or tryptophan hydrochloride are generally directly available) is subjected to a Pictet-Spengler reaction to give (R) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole-3-carboxylic acid, which is esterified in the presence of thionyl chloride and methanol to form methyl (R) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole-3-carboxylate in the C-3 position; the compound reacts with chloracetyl chloride to obtain an amide product (R) -2- (2-chloracetyl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole-3-carboxylic acid methyl ester; the compound is finally refluxed in a methanol solution of methylamine to obtain a compound 12;

preferably, the reaction can be carried out by conventional separation means such as column chromatography, recrystallization and the like to obtain pure products of the reaction in each step;

preferably, the use of (L) -tryptophan or a racemic tryptophan mixture enables the (S) -enantiomer or racemate, respectively, of compound 12 to be obtained;

preferably, the compound 12 is prepared by the following reaction:

preferably, the compound 14 is prepared by the following reaction:

(D) -tryptophan or tryptophan hydrochloride by a Pictet-Spengler reaction to give (R) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole-3-carboxylic acid, which is esterified in the presence of thionyl chloride and methanol to form methyl (R) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole-3-carboxylate in the C-3 position; the compound reacts with isocyanate to generate (R) -2- (ethylcarbamoyl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole-3-carboxylic acid methyl ester, and the methyl ester is refluxed in the presence of cesium carbonate to obtain a compound 14;

preferably, the compound 14 is prepared by the following reaction:

preferably, the compound 17 is prepared by the following reaction:

the tryptamine hydrochloride is taken as a starting material, and corresponding β -tetrahydrocarboline is obtained through Pictet-Spengler reaction, and the compound is converted into a compound 17 through coupling reaction;

preferably, the reaction is carried out by conventional separation means such as column chromatography, recrystallization and the like to obtain a pure product of the reaction;

preferably, the compound 17 is prepared by the following reaction:

preferably, in the above-mentioned several reactions, X represents a sulfur atom or an oxygen atom; r₁Represents a hydrogen atom, a bromine atom and C₁-C₂The position of the substituent is C-3 or C-5, R₁And a furyl group may also represent a benzofuryl group; r₂Represents C₁-C₂Alkyl groups of (a); r₃Represents bipyridyl or 4- (3,4 dimethoxyphenyl) pyrimidine; r₄Represents a bromine atom, a methyl formate group or a formic acid group.

Preferably, the oxidizing agent TEMPO tetrafluoroborate is prepared as described in J.Med.chem.2016,59, 5063-5076; for preparing the structure

The main methods for potassium fluoborate salt compounds can be referred to in eur.j.med.chem.2018,150, 30-38. Wherein, the pyrazine ring structure (compound 12) is used as a raw material, the reaction yield is 10 to 25 percent, andthe ratio trans of enantiomer products, cis ═ 1.8-3.3: 1; using the acetyl urea structure (compound 14) as the raw material, the reaction yield is 18-35%, and the diastereoisomer product ratio trans: cis>20: 1. the yield can reach 70-90% by taking the derivative of N-aryl or N-amide (compound 17) as the starting material.

preferably, the (S) -enantiomer or racemate of the compound can be obtained using (L) -tryptophan or a racemic tryptophan mixture, respectively.

For example, D-tryptophan methyl ester (2g,10mmol,1eq) and 5-ethylfurfural were dissolved in 100mL of chloroform, 80 drops of trifluoroacetic acid were slowly added, the reaction was worked up after stirring at 60 ℃ for 10 hours, column chromatography was carried out to obtain 1- (5-ethylfuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole-3-carboxylic acid methyl ester, a cis-trans isomer mixture, and the cis-trans isomer mixture was dissolved in 3mL of dichloromethane. DIEA (20. mu.L, 0.11mmol,1.1eq) and ethyl isocyanate (10. mu.L, 1.3mmol,1.3eq) were added to the solution, and the mixture was reacted at room temperature for 3 hours. Purification by column chromatography gives the compound of formula VII (X ═ O).

Also provided herein are compositions or formulations comprising a compound as described above, or an optically pure enantiomer or solvate or hydrate thereof, or a pharmaceutically acceptable salt thereof; preferably, the dosage forms include tablets, capsules, lozenges, dragees, pills, granules, powders, solutions, emulsions, suspensions, dispersions, syrups, gels, aerosols and other special dosage forms; preferably, the composition or formulation includes a pharmaceutical composition or formulation for human use and a pharmaceutical composition or formulation for veterinary use.

The application also provides the use of the above compound or its optically pure enantiomer or its solvate or its hydrate or its pharmaceutically acceptable salt or the above pharmaceutical composition or preparation for the preparation of a medicament for treating, alleviating or preventing a disease or condition associated with PDE5 in a mammal; preferably, the treatment, alleviation or prevention is manifested by inhibition of phosphodiesterase type 5 by the above-mentioned compound or its optically pure enantiomer or its solvate or its hydrate or its pharmaceutically acceptable salt or the above-mentioned pharmaceutical composition or preparation, thereby eliminating, ameliorating or reducing the disease or condition associated with PDE 5; preferably, the use of a compound as described above or an optically pure enantiomer or solvate or hydrate thereof or a pharmaceutically acceptable salt or hydrate thereof or a pharmaceutical composition or formulation as described above for the preparation of a PDE5 inhibitor.

The application provides the use of the compound or the optically pure enantiomer or the solvate or the hydrate of the enantiomer or the pharmaceutically acceptable salt of the enantiomer or the hydrate of the solvate or the pharmaceutically acceptable salt of the enantiomer or the pharmaceutically acceptable salt of the pharmaceutical composition or the preparation for preparing a medicament for treating, relieving or preventing vascular-related diseases or disease states in mammals; preferably, the treatment, alleviation or prevention is characterized in that the compound or the optically pure enantiomer or the solvate or the hydrate of the enantiomer or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition or the preparation improves local arterial blood flow or reduces vascular pressure, thereby eliminating, improving or alleviating the vascular-related disease or condition.

Preferably, the disease or condition is selected from, but not limited to, Male Erectile Dysfunction (MED), male testicular ischemia, Female Sexual Dysfunction (FSD), premature labor, dysmenorrhea, Benign Prostatic Hyperplasia (BPH), bladder outlet obstruction, incontinence, stable, unstable and variant (prinzmetal's) angina, hypertension, pulmonary Hypertension (HP), alzheimer's disease (senile dementia, AD), congestive heart failure, arteriosclerosis, stroke, peripheral vascular disease, reduced vascular patency, chronic asthma, bronchitis, allergic asthma, allergic rhinitis, glaucoma, or a disease characterized by bowel motility disorders; preferably, the disease or condition is selected from Male Erectile Dysfunction (MED) and Pulmonary Arterial Hypertension (PAH); preferably, the above compound or an optically pure enantiomer or a solvate or hydrate thereof or a pharmaceutically acceptable salt thereof or the above pharmaceutical composition or formulation is administered in a manner selected from oral administration, topical administration, intramuscular administration, intradermal administration, intraperitoneal administration, subcutaneous administration, intravenous administration and inhalation administration, preferably oral administration.

The various terms and phrases used herein have the ordinary meaning as is known to those skilled in the art, and even then, it is intended that the present invention not be limited to the specific meanings and expressions employed herein as are inconsistent with such known meanings. The following are definitions of various terms used herein, which apply to the terms used throughout this specification unless otherwise specified in specific instances.

In the context of the present invention, a radical, as depicted for example in a compound of the general formula or a specific compound, may link several hydrogen atoms, so that the radical may satisfy the chemical valence requirements, although the hydrogen atoms on the corresponding radical may not be depicted in the structural formula.

In the compounds of general formula X herein, when a substituent is attached to the interior of a ring via a bond, it means that the substituent may be substituted at any position of the ring that may be substituted, for example for R₁Substituents and the ring in which they are located wherein one or more R₁Substituents may be located at the 2-, 3-, 4-, 5-positions of the ring.

As used herein, the term "pharmaceutical composition" may also refer to a "composition" that may be used to effect treatment or prevention of a disease or disorder described herein in a subject, particularly a mammal.

As used herein, the term "pharmaceutically acceptable" or "pharmaceutically acceptable" used interchangeably therewith, such as in the description of "pharmaceutically acceptable salts", means that the salts are not only physiologically acceptable to the subject, but also synthetic substances of value in pharmaceutical use, such as salts formed as intermediates in the performance of chiral separations, which salts, although not directly administered to the subject, may play a role in obtaining the final product of the invention.

Pharmaceutical compositions of the compounds of the present invention may be administered in any of the following ways: oral, aerosol inhalation, rectal, nasal, vaginal, topical, parenteral such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial injection or infusion, or by means of an explanted reservoir, with oral, intramuscular, intraperitoneal or intravenous administration being preferred.

The compounds of the present invention or pharmaceutical compositions containing them may be administered in unit dosage form. The administration dosage form can be liquid dosage form or solid dosage form. The liquid dosage form can be true solution, colloid, microparticle, emulsion, or mixed suspension. Other dosage forms such as tablet, capsule, dripping pill, aerosol, pill, powder, solution, suspension, emulsion, granule, suppository, lyophilized powder for injection, clathrate, landfill, patch, liniment, etc.

The pharmaceutical compositions of the present invention may also contain conventional carriers, including but not limited to: ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycerol, sorbates, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, beeswax, lanolin and the like. The carrier may be present in the pharmaceutical composition in an amount of 1% to 98% by weight, typically about 80% by weight. For convenience, the local anesthetic, preservative, buffer, etc. may be dissolved directly in the vehicle.

Oral tablets and capsules may contain excipients such as binding agents, for example syrup, acacia, sorbitol, tragacanth, or polyvinylpyrrolidone, fillers such as lactose, sucrose, corn starch, calcium phosphate, sorbitol, glycine, lubricants such as magnesium stearate, talc, polyethylene glycol, silica, disintegrants such as potato starch, or acceptable wetting agents such as sodium lauryl sulfate. The tablets may be coated by methods known in the art of pharmacy.

The oral liquid can be made into water and oil suspension, solution, emulsion, syrup, or dried product, and supplemented with water or other suitable medium before use. Such liquid preparations may contain conventional additives such as suspending agents, sorbitol, cellulose methyl ether, glucose syrup, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gelatin, hydrogenated edible fats and oils, emulsifying agents such as lecithin, sorbitan monooleate, gum arabic; or a non-aqueous carrier (which may comprise an edible oil), such as almond oil, an oil such as glycerol, ethylene glycol, or ethanol; preservatives, e.g. methyl or propyl p-hydroxybenzoates, sorbic acid. Flavoring or coloring agents may be added if desired.

Suppositories may contain conventional suppository bases such as cocoa butter or other glycerides.

For parenteral administration, liquid dosage forms are generally prepared from the compound and a sterile carrier. The carrier is preferably water. The compound can be dissolved in the carrier or made into suspension solution according to the concentration of the carrier and the drug, and the compound is firstly dissolved in water when made into the solution for injection, filtered and sterilized and then filled into a sealed bottle or ampoule.

It will be appreciated that the optimum dosage and interval for administration of a compound of formula (I) will be determined by the nature of the compound and external conditions, such as the form, route and route of administration and the particular mammal being treated, and that such optimum dosage may be determined by conventional techniques. It should also be recognized that the optimal course of treatment, i.e., the daily dosage of compound X at the same time over the nominal time, may be determined by methods well known in the art.

Drawings

Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:

FIG. 1 shows the results of an experiment on the inhibitory effect of compound II-2 on PED-5 at a concentration of 100 nM;

FIG. 2 is a graph of the effect of compounds I-9, II-2, I-13 on the concentration-dependent relaxation of arterioles, the top-down three lines of the graph showing cumulative increase with dose (10) for compounds I-9, II-2, I-13, respectively^-11To 10^-5M) changes in vessel caliber.

Detailed Description

The following examples further illustrate the synthesis and preparation methods of the compounds and intermediates of the present invention, and the research methods of the in vitro phosphodiesterase inhibitory activity and vasodilation activity of the compounds. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

Example 1Having a heterocyclic ring structure

Preparation of potassium trifluoroborate

Wherein X is oxygen or sulfur, R₁Is hydrogen, bromine, methyl or ethyl, R₁The substitution position of (A) is C-3 or C-5; when X ═ O, R₁The combination with a furanyl group may be a benzofuran.

Preparation of potassium 5-ethylfuran-2-trifluoroborate

To a round bottom flask was added 2-ethylfuran (1.9g,20mmol,1eq) sequentially; and 15mL of tetrahydrofuran. Under nitrogen protection, 10mL of an n-butyllithium solution (2.5M) and a hexane solution were added dropwise, and after stirring for 3 hours, the mixture was added dropwise to trimethyl borate (2.28g,22mmol,1.1eq) in tetrahydrofuran (5 mL). After the completion of the dropwise addition, the temperature was returned to room temperature, the mixture was stirred for 0.5 hour, then cooled to 0 ℃ again, and 20mL of 1M hydrochloric acid was added dropwise to a pH of 5 to 6. Extracting with ethyl acetate, evaporating to dryness, and recrystallizing to obtain 1.25g of pure 5-ethylfuran-2-boronic acid. 5-Ethylfuran-2-boronic acid (610mg,4.35mmol,1eq), KHF was added to a round bottom flask at 0 deg.C₂(1.1g,13.05mmol,3 eq); methanol 1.5mL, water 3.8 mL. The reaction was stirred for 10min, evaporated to dryness and pumped to dryness under high vacuum overnight. Adding acetone into the solid, refluxing for 0.5h, filtering, evaporating the filtrate to dryness, and usingEther recrystallized potassium 5-ethylfuran-2-trifluoroborate, 460mg, 23% yield over 2 steps.¹H NMR(400MHz,DMSO)δ5.89(d,J＝2.8Hz,1H),5.74(d,J＝2.3Hz,1H),2.60–2.51(m,2H),1.14(t,J＝7.5Hz,3H).

Example 2Preparation of Compounds of formula I

Scheme a:

Preparation of Compound I-3 and diastereomers thereof

1. Preparation of (R) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6] pyrido [3,4-b ] indole-1, 4-dione (Compound 12)

Dissolving D-tryptophan (2.04g,10mmol,1eq) in 30mL of water, adding NaOH (440mg,11mmol,1.1eq) and 40% formaldehyde solution (860 mu L,11mmol,1.1eq), stirring at room temperature for 2H, refluxing for 3H, neutralizing with acetic acid, precipitating, and filtering to obtain the corresponding product, (R) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ]]Indole-3-carboxylic acid (Compound 9), 1.5 g.¹HNMR(400MHz,DMSO-d₆)δ10.92(s,1H),7.44(d,J＝7.9Hz,1H),7.33(d,J＝8.0Hz,1H),7.07(t,J＝7.5Hz,1H),6.99(t,J＝7.5Hz,1H),4.26–4.11(m,2H),3.60(dd,J＝10.2,4.9Hz,2H),3.17(s,5H),3.14–3.10(m,1H),2.81(dd,J＝15.8,10.2Hz,1H).m/z 215[M-H]^-。

Compound 9(434mg,2mmol,1eq) was dissolved in 37mL MeOH, thionyl chloride (416. mu.L, 3mmol,1.5 eq) was added at 0 deg.C, stirred for 2h and then refluxed for 15h over Na₂CO₃Neutralizing, extracting with ethyl acetate, evaporating to dryness, and recrystallizing to obtain product (R) -2,3,4, 9-tetrahydro-1H-pyrido [3, 4-b)]Indole-3-carboxylic acid methyl ester (compound 10), 420 mg.¹H NMR(400MHz,DMSO-d₆)δ10.71(s,1H),7.37(d,J＝7.7Hz,1H),7.26(d,J＝8.0Hz,1H),7.08–6.97(m,1H),6.93(t,J＝7.3Hz,1H),4.09–3.84(m,2H),3.73(dd,J＝8.9,4.7Hz,1H),2.93(dd,J＝15.1,4.7Hz,1H),2.85(s,1H),2.74(dd,J＝14.9,8.7Hz,1H).m/z 231[M+H]⁺.

The compound 10(1g,4.3mmol,1eq) was dissolved in anhydrous dichloromethane, chloroacetyl chloride (380 μ L,4.8mmol,1.1eq) and triethylamine (1.2mL,8.7mmol,2eq) were added to react at room temperature for 3 hours, water was added to terminate the reaction, and the pure compound (R) -2- (2-chloroacetyl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole-3-carboxylic acid methyl ester (compound 11), 880mg was obtained by column chromatography after extraction.

Directly dissolving the compound 11(500mg,1.3mmol,1eq) in a methanol solution of methylamine (274mg,6.8mmol,5eq), refluxing for 3h, evaporating to dryness, and performing column chromatography to obtain the product (R) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6]Pyridine [3,4-b ]]Indole-1, 4-dione (Compound 12), 350 mg.¹H NMR(400MHz,DMSO-d₆)δ10.96(s,1H),7.43(d,J＝7.7Hz,1H),7.33(d,J＝8.0Hz,1H),7.07(t,J＝6.9Hz,1H),6.98(t,J＝6.9Hz,1H),5.37(dd,J＝16.5,1.4Hz,1H),4.32(dd,J＝11.7,4.2Hz,1H),4.19(dd,J＝17.3,3.6Hz,2H),4.02(d,J＝17.6Hz,1H),3.20(dd,J＝15.2,4.3Hz,1H),2.90(s,3H),2.88–2.78(m,1H).m/z 270[M+H]⁺.

2. Preparation of Compound I-3 and diastereomers thereof

Compound 12(135mg,0.5mmol,1eq), potassium 5-ethylfuran-2-trifluoroborate (310mg,1.5mmol,3eq) was dissolved in 40mL of 1:1 DCM/CHCl₃To the combined solution, TEMPO (500mg,2mmol,4eq) was added in four portions. The reaction is stirred for 3h, then water is added for treatment, and the corresponding product I-3 and diastereoisomer thereof, namely (6S,12aR) -6- (5-ethyl furan-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazine [1',2':1,6]Pyridine [3,4-b ]]Indole-1, 4-diones (Compound I-3) and (6R,12aR) -6- (5-ethylfuran-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6]Pyridine [3,4-b ]]Indole-1, 4-dione, 8mg and 15mg, respectively.

Scheme B:

dissolving D-tryptophan methyl ester (2g,10mmol,1eq) and 5-ethylfurfural in 100mL of chloroform, slowly adding 80 drops of trifluoroacetic acid, stirring at 60 ℃ for 10H, performing post-treatment reaction, and performing column chromatography to obtain 1- (5-ethylfuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] -pyridine]Indole-3-carboxylic acid methyl ester, cis-trans-isoMixture M1, total 750mg, M/z 325[ M + H ]]⁺. Dissolving the mixture M1(590mg,1.9mmol,1eq) in anhydrous dichloromethane, adding chloroacetyl chloride (200. mu.L, 2.28mmol,1.2eq) and triethylamine (0.5mL,3.8mmol,2eq), reacting at room temperature for 3H, adding water to terminate the reaction, extracting, and separating by column chromatography to obtain 2- (2-chloroacetyl) -1- (5-ethylfuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ]]Indole-3-carboxylic acid methyl ester, cis-trans isomer mixture M2, total 550mg, M/z 401[ M + H ]]⁺. Dissolving the mixture M2 in methylamine methanol solution, refluxing for 3 hr, evaporating to dryness, and performing column chromatography to obtain product I-3 and its diastereoisomer, 280mg and 116mg respectively.

(6S,12aR) -6- (5-Ethylfuran-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6]Pyridine [3,4-b ]]Indole-1, 4-dione (Compound I-3) as colorless crystal, mp.221-223 ℃.¹H NMR(400MHz,DMSO-d₆)δ11.20(s,1H),7.58(d,J＝7.8Hz,1H),7.34(d,J＝8.0Hz,1H),7.08(t,J＝7.6Hz,1H),7.01(t,J＝7.1Hz,1H),6.36(s,1H),6.05(d,J＝3.1Hz,1H),5.91(d,J＝3.1Hz,1H),4.32(dd,J＝11.8,4.4Hz,1H),4.20(dd,J＝17.1,1.5Hz,1H),3.96(d,J＝17.0Hz,1H),3.50(dd,J＝15.4,4.6Hz,1H),2.93(s,4H),2.45(q,J＝7.5Hz,2H),1.05(t,J＝7.5Hz,3H).¹³CNMR(101MHz,DMSO)δ167.3,167.2,156.6,151.8,136.4,131.8,126.1,121.7,119.4,118.6,112.0,107.2,106.7,105.2,55.8,52.0,49.0,33.4,22.9,21.1,12.1.ESI-MS m/z 364[M+H]⁺,HR-EI-MS:m/z 364.1657[M+H]⁺,(calcd for C₂₁H₂₂N₃O₃ ⁺,364.1656).

(6R,12aR) -6- (5-Ethylfuran-2-yl) -2-methyl-2, 3,6,7,12,12 a-hexahydropyrazino [1',2':1,6]Pyridine [3,4-b ]]Indole-1, 4-dione, colorless crystals, mp.230-232 ℃.¹H NMR(400MHz,DMSO-d₆)δ11.11(s,1H),7.49(d,J＝7.8Hz,1H),7.32(d,J＝8.1Hz,1H),7.11(t,J＝7.5Hz,1H),7.01(t,J＝7.4Hz,1H),6.85(s,1H),6.10(d,J＝3.1Hz,1H),6.05(d,J＝3.1Hz,1H),4.39–4.16(m,2H),4.08(d,J＝17.8Hz,1H),3.23(dd,J＝15.4,4.2Hz,1H),2.97(dd,J＝15.4,11.8Hz,1H),2.88(s,3H),2.58(q,J＝7.5Hz,2H),1.13(t,J＝7.5Hz,3H).¹³C NMR(101MHz,DMSO)δ165.2,163.0,158.2,149.8,136.6,129.5,126.3,122.2,119.3,118.6,111.8,110.9,107.5,105.5,53.3,51.1,46.2,33.1,26.9,21.2,12.3.ESI-MS m/z 364[M+H]⁺,HR-EI-MS:m/z 364.1658[M+H]⁺,(calcd for C₂₁H₂₂N₃O₃ ⁺,364.1656).

Other compounds of formula I may be prepared by selecting appropriate substituents according to the above synthetic route and following the methods for the preparation of compound I-3 and its diastereoisomers, respectively.

Example 3Preparation of Compounds of formulae II, VII

Wherein X is oxygen or sulfur; r₁Is hydrogen, bromine, methyl or ethyl, R₁The position of the substituent is C-3 or C-5; when X is O, R₁And a furyl group may also represent a benzofuryl group; r₂Is methyl or ethyl.

Preparation of Compounds II-2 and II-3

1. Preparation of Compound 14

Enantiomer (S) -10 of compound 10 was prepared by the method of reference example 2. To a round bottom flask were added (S) -10(460mg,2mmol,1eq), ethyl isocyanate (265. mu.L, 3.2mmol,1.6eq), and 20mL of 2-butanone, and the reaction was stirred under reflux for 3h and worked up. Performing column chromatography to obtain (S) -2- (ethylcarbamyl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b]Indole-3-carboxylic acid methyl ester (compound 13), 430 mg.¹H NMR(400MHz,Acetone-d₆)δ10.01(s,1H),7.46(d,J＝7.8Hz,1H),7.37–7.27(m,1H),7.10–7.04(m,1H),7.01(td,J＝7.4,1.1Hz,1H),6.13(s,1H),5.58–5.48(m,1H),4.82(dd,J＝15.0,1.9Hz,1H),4.62(dt,J＝15.1,1.9Hz,1H),3.55(s,3H),3.40(dt,J＝15.6,1.6Hz,1H),3.32–3.21(m,2H),3.04(ddt,J＝15.3,6.2,1.9Hz,1H),1.11(t,J＝7.2Hz,3H).m/z 302[M+H]⁺

The purified compound 13(150mg,0.5mmol,1eq) was dissolved in 5mL of 1, 4-dioxane, and Cs was added₂CO₃(325mg,1mmol,2eq), stirring at 90 ℃ for reaction for 1H, evaporating to dryness, carrying out column chromatography to obtain corresponding (S) -2-ethyl-5, 6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6]pyridine [3,4-b ]]Indole-1, 3(2H) -dione (Compound 14),121 mg. 1H NMR (400MHz, Acetone-d₆)δ10.16(s,1H),7.51(d,J＝7.8Hz,1H),7.39(d,J＝8.0Hz,1H),7.20–6.98(m,2H),5.03(dd,J＝16.2,1.8Hz,1H),4.43(dt,J＝16.3,1.8Hz,1H),4.32(ddd,J＝11.1,5.5,2.5Hz,1H),3.54(q,J＝7.2Hz,2H),3.29(ddd,J＝14.9,5.5,1.5Hz,1H),2.75(ddd,J＝15.1,10.9,2.1Hz,1H),1.18(t,J＝7.2Hz,3H).m/z 270[M+H]⁺

2. Preparation of Compound II-2

Compound 14(26.9mg,0.1mmol,1eq), potassium 5-ethylfuran-2-trifluoroborate (41mg,0.2mmol,2eq) was dissolved in 3mL of a 1:1 mixed solution of DCM, and TEMPO (75mg,0.3mmol,3eq) was added in three portions. The reaction was stirred for 1h, then treated with water and chromatographed to give the corresponding product II-2, 8 mg.

Alternatively, methyl 1- (5-ethylfuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole-3-carboxylate, a mixture of cis-trans isomers, was prepared starting with L-tryptophan methyl ester in the same manner as in reference example 2. The mixture (410mg,1.4mmol,1eq) was dissolved in 2-butanone, ethyl isocyanate was added and refluxed for 8h, then the column chromatography was evaporated to dryness to obtain compounds II-2 and II-3, 220mg and 84mg respectively.

(5S,11aS) -2-Ethyl-5- (5-ethylfuran-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6]Pyridine [3,4-b ]]Indole-1, 3(2H) -dione (Compound II-2), colorless crystals, mp.150-152 ℃.¹H NMR(400MHz,DMSO-d₆)δ11.04(s,1H),7.52(d,J＝7.8Hz,1H),7.33(d,J＝8.0Hz,1H),7.11(t,J＝7.5Hz,1H),7.02(t,J＝7.4Hz,1H),6.24(t,J＝2.6Hz,2H),6.05(d,J＝3.1Hz,1H),4.49(dd,J＝10.8,5.7Hz,1H),3.47(q,J＝7.2Hz,2H),3.33(dd,J＝15.2,5.8Hz,1H),2.80(ddd,J＝15.2,10.9,1.8Hz,1H),2.56(q,J＝7.6Hz,2H),1.17–1.07(m,6H).¹³C NMR(101MHz,DMSO)δ172.8,157.8,154.4,150.5,137.0,130.0,126.2,122.2,119.3,118.7,111.9,109.2,106.2,105.5,53.5,45.8,33.5,23.1,21.2,13.7,12.2.ESI-MS m/z 364[M+H]⁺,HR-EI-MS:m/z 364.1657[M+H]⁺,(calcd for C₂₁H₂₂N₃O₃ ⁺,364.1656).

(5R,11aS) -2-ethyl-5- (5-ethylfuran-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6]Pyridine [3,4-b ]]Indole-1, 3(2H) -dione (Compound II-3), pale yellow crystals, mp.95-96 ℃.¹H NMR(400MHz,DMSO-d₆)δ10.84(s,1H),7.53(d,J＝7.7Hz,1H),7.28(d,J＝8.0Hz,1H),7.07(t,J＝7.7Hz,1H),7.00(t,J＝7.4Hz,1H),6.33(d,J＝3.1Hz,1H),6.03(d,J＝3.1Hz,1H),5.98(s,1H),4.49(dd,J＝11.4,4.5Hz,1H),3.41(q,J＝7.2Hz,2H),3.28(dd,J＝14.7,4.6Hz,1H),2.82(ddd,J＝13.9,11.5,1.9Hz,1H),2.49(m,2H,overlapped with DMSO),1.09(t,J＝7.4Hz,6H).¹³C NMR(101MHz,DMSO)δ171.9,157.0,154.2,149.2,136.9,132.2,126.3,122.0,119.3,118.6,111.9,109.7,106.3,105.5,57.9,49.8,33.2,22.4,21.2,13.8,12.2.ESI-MS m/z 364[M+H]⁺,HR-EI-MS:m/z364.1657[M+H]⁺,(calcd for C₂₁H₂₂N₃O₃ ⁺,364.1656).

Preparation of Compound VII

With reference to the above conditions, the reaction temperature is lowered and the solvent is replaced and then used to prepare compound VII. 1- (5-Ethylfuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b]The mixture of methyl indole-3-carboxylate, cis-trans isomer was dissolved in 3mL of dichloromethane. DIEA (20. mu.L, 0.11mmol,1.1eq) and ethyl isocyanate (10. mu.L, 1.3mmol,1.3eq) were added to the solution, and the mixture was reacted at room temperature for 3 hours. After column chromatography purification, the corresponding product 5mg, methyl (1S,3R) -2- (ethylcarbamoyl) -1- (5-ethylfuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] is obtained]Indole-3-carboxylic ester (VII), yellow crystal, mp.93-95 deg.C.¹H NMR(400MHz,DMSO-d₆)δ10.92(s,1H),7.49(d,J＝7.8Hz,1H),7.30(d,J＝8.1Hz,1H),7.09(t,J＝7.4Hz,1H),7.00(t,J＝7.4Hz,1H),6.79(t,J＝5.3Hz,1H),6.50(s,1H),5.95(d,J＝3.0Hz,1H),5.68(d,J＝3.1Hz,1H),5.39(d,J＝6.6Hz,1H),3.37(s,1H),3.28–3.07(m,5H),2.82(dd,J＝15.7,6.8Hz,1H),2.57(q,J＝7.6Hz,2H),1.15(t,J＝7.5Hz,3H),1.09(t,J＝7.1Hz,3H).ESI-MS m/z 396[M+H]⁺,HR-EI-MS:m/z396.1913[M+H]⁺,(calcd for C₂₂H₂₆N₃O₄ ⁺,396.1918).

Preparation of Compound II-12

Compounds were prepared with reference to the above procedure: (5R,11 a)R) -2-ethyl-5- (5-ethylthiophen-2-yl) -5,6,11,11 a-tetrahydro-1H-imidazo [1',5':1,6]Pyridine [3,4-b ]]Indole-1, 3(2H) -dione (Compound II-7). Compound II-7(104mg,0.22mmol,1eq) and DDQ (98mg,0.43mmol,2eq) powder were cooled to-78 deg.C and mixed well, H was slowly added dropwise₂O/THF (7.2/0.8 mL). Stirring for 1h, heating to room temperature, stirring for 2h, and adding Na₂CO₃The reaction was terminated. Performing column chromatography to obtain product, 25mg, N-ethyl-1- (5-ethylthiophene-2-yl) -9H-pyrrole [3,4-b]Indole-3-carboxamide (Compound II-12), light yellow crystals, mp.101-103 ℃.¹H NMR(400MHz,DMSO-d₆)δ11.74(s,1H),8.75(s,1H),8.52–8.26(m,2H),8.00(d,J＝3.8Hz,1H),7.76(d,J＝8.2Hz,1H),7.62(t,J＝7.7Hz,1H),7.33(t,J＝7.5Hz,1H),7.10(d,J＝3.7Hz,1H),3.51–3.39(m,2H),2.94(q,J＝7.5Hz,2H),1.36(t,J＝7.5Hz,3H),1.21(t,J＝7.0Hz,3H).¹³C NMR(101MHz,DMSO)δ164.6,150.2,142.0,139.8,139.8,135.6,132.4,130.7,129.2,127.2,125.7,122.5,121.6,120.9,113.3,112.9,34.2,23.5,16.3,15.6.ESI-MS m/z 378[M+H]⁺,HR-EI-MS:m/z 350.1323[M+H]⁺,(calcd for C₂₀H₂₀N₃OS⁺,350.1322).

Example 4Preparation of Compounds of formula III

Wherein X is sulfur or oxygen; r₁Is hydrogen, bromine, methyl or ethyl, R₁The position of the substituent is C-3 or C-5; when X is O, R₁And a furyl group may also represent a benzofuryl group; r₃Is bipyridyl or 4- (3,4 dimethoxyphenyl) pyrimidine; r₄Is a bromine atom, a methyl formate group or a formic acid group.

Preparation of Compound III-1

Reference eur.j.med.chem.2017,150,30-38 preparation of 2- (5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b]Indole as starting material was added to a round bottom flask (386mg,1mmol,1eq), followed by the addition of potassium 5-ethylfuran-2-trifluoroborate (327mg,1.5mmol,1.5eq), and dichloroMethane 20mL, and TEMPO (243mg,1mmol,1 eq). After stirring at room temperature for 30min, saturated Na was added₂CO₃The solution was terminated, the organic phase was separated, washed once with saturated NaCl, and the layers were separated. The organic phase was dried over anhydrous MgSO4, filtered, and concentrated to dryness. Separating after column chromatography to obtain 310mg of pure product. 2- (5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) -1- (5-ethylfuran-2-yl) -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b]Indole (Compound III-1), colorless crystals, mp87-89 ℃.¹H NMR(400MHz,CDCl₃)δ8.56(s,2H),8.15(s,1H),7.54(d,J＝7.6Hz,1H),7.32(d,J＝8.0Hz,1H),7.20–7.07(m,3H),7.01(dd,J＝8.2,2.1Hz,1H),6.97–6.89(m,2H),6.08(d,J＝3.1Hz,1H),5.87(d,J＝3.1Hz,1H),5.13(dd,J＝13.3,4.8Hz,1H),3.92(s,3H),3.90(s,3H),3.65–3.45(m,1H),3.13–2.89(m,1H),2.84(dd,J＝15.4,3.2Hz,1H),2.59(q,J＝7.5Hz,2H),1.17(t,J＝7.6Hz,3H).¹³C NMR(101MHz,CDCl₃)δ160.5,158.1,155.8,151.2,149.5,148.6,136.2,131.3,128.6,126.9,123.4,122.0,119.5,118.4,118.3,111.8,111.0,110.4,109.3,108.5,104.4,56.0,56.0,49.2,38.8,21.4,21.0,12.0.ESI-MS m/z 481[M+H]⁺,HR-EI-MS:m/z 481.2230[M+H]⁺,(calcd for C₂₉H₂₉N₄O₃ ⁺,481.2234).

Other compounds of formula III may be prepared by selecting the corresponding substituent groups according to the above synthetic route and following the procedure for preparing compound III-1.

Example 5Preparation of Compounds of formula V, IV

Preparation of Compound IV-1 and Compound V-1

Referring to the procedure of example 4, starting from compound III-1. III-1(104mg,0.22mmol,1eq) and DDQ (98mg,0.43mmol,2eq) powders were cooled to-78 deg.C and mixed well, H was slowly added dropwise₂O/THF (7.2/0.8 mL). Stirring for 1h, heating to room temperature, stirring for 2h, and adding Na₂CO₃The reaction was terminated. After column chromatography, the compound IV-1 and the compound V-1 are obtained, which are 23mg and 51mg respectively. (3- (2- ((5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) amino) ethyl) -1H-indol-2-yl) (5-ethylfuran-2-yl) methanone (IV-1) as yellow crystals, mp.121-123 ℃.¹H NMR(400MHz,Methanol-d₄)δ8.62(s,2H),8.10(dd,J＝5.9,2.8Hz,1H),7.50(s,1H),7.47–7.41(m,1H),7.31–7.22(m,2H),7.10–7.07(m,1H),7.05(dd,J＝8.3,2.2Hz,1H),6.97(d,J＝8.3Hz,1H),6.12(d,J＝3.2Hz,1H),5.93(d,J＝3.0Hz,1H),5.35(d,J＝18.1Hz,1H),4.02(d,J＝18.1Hz,1H),3.86(s,3H),3.83(s,3H),2.56–2.49(m,2H),1.11(t,J＝7.6Hz,3H).¹³C NMR(101MHz,MeOD)δ190.0,159.4,158.3,155.4,149.6,149.0,148.9,148.3,136.8,127.9,124.6,123.8,123.7,122.4,120.8,118.2,112.0,111.7,111.0,109.4,109.2,104.4,55.1,55.0,49.3,20.7,11.0.ESI-MS m/z 495[M+H]⁺,HR-EI-MS:m/z497.2179[M+H]⁺,(calcd for C₂₉H₂₉N₄O₄ ⁺497.2183) 2- (5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) -1- (5-ethylfuran-2-yl) -1,2,3, 9-tetrahydro-4H-pyridine [3,4-b ]]Indol-4-one (V-1), yellow crystals, mp.121-123 ℃.¹H NMR(400MHz,Methanol-d₄)δ8.62(s,2H),8.10(dd,J＝5.9,2.8Hz,1H),7.50(s,1H),7.47–7.41(m,1H),7.31–7.22(m,2H),7.10–7.07(m,1H),7.05(dd,J＝8.3,2.2Hz,1H),6.97(d,J＝8.3Hz,1H),6.12(d,J＝3.2Hz,1H),5.93(d,J＝3.0Hz,1H),5.35(d,J＝18.1Hz,1H),4.02(d,J＝18.1Hz,1H),3.86(s,3H),3.83(s,3H),2.56–2.49(m,2H),1.11(t,J＝7.6Hz,3H).¹³C NMR(101MHz,MeOD)δ190.0,159.4,158.3,155.4,149.6,149.0,148.9,148.3,136.8,127.9,124.6,123.8,123.7,122.4,120.8,118.2,112.0,111.7,111.0,109.4,109.2,104.4,55.1,55.0,49.3,20.7,11.0.ESI-MS m/z 495[M+H]⁺,HR-EI-MS:m/z495.2019[M+H]⁺,(calcd for C₂₉H₂₇N₄O₄ ⁺,495.2027).

Example 6Preparation of Compounds of formula VI

Preparation of Compound VI

Reference eur.j.med.chem.2018,150,30-38 preparation of methyl- (E) -4- (3-oxo- (1,3,4, 9-tetrahydro-2H-pyridine [3,4-b ]]Indol-2-yl) -1-en-1-yl) benzoate was the starting material. This starting material (360mg,1mmol,1eq) was dissolved in 20mL of dichloromethane, and potassium 5-ethylthiophene-2-trifluoroborate (327mg,1.5mmol,1.5eq), 20mL of dichloromethane, and TEMPO (243mg,1mmol,1eq) were added. After stirring at room temperature for 30min, saturated Na was added₂CO₃The solution was terminated, the organic phase was separated, washed once with saturated NaCl, and the layers were separated. The organic phase was dried over anhydrous MgSO4, filtered, and concentrated to dryness. After column chromatography, 420mg of methyl ester product is obtained. Methyl- (E) -4- (3- (1- (5-ethylthiophen-2-yl) -1,3,4, 9-tetrahydro-2H-pyridine [3, 4-b)]Indol-2-yl) -3-oxo-1-en-1-yl benzoate (compound VI-2) as colorless crystals, mp.115-117 ℃.¹H NMR(400MHz,DMSO-d₆)δ11.09(s,1H),7.98(d,J＝8.3Hz,2H),7.93(d,J＝8.2Hz,2H),7.65(d,J＝15.2Hz,1H),7.53(d,J＝15.4Hz,1H),7.48(d,J＝7.9Hz,1H),7.31(d,J＝8.1Hz,1H),7.09(t,J＝7.6Hz,1H),7.01(t,J＝7.6Hz,1H),6.97(s,1H),6.69(d,J＝3.5Hz,1H),6.66(d,J＝3.5Hz,1H),4.53(d,J＝13.9Hz,1H),3.87(s,3H),3.55–3.38(m,1H),2.94–2.79(m,2H),2.75(q,J＝7.7Hz,2H),1.19(t,J＝7.4Hz,3H).¹³CNMR(101MHz,DMSO)δ166.3,164.8,147.5,141.5,140.6,140.1,136.6,132.1,130.5,129.9,128.9,127.1,126.4,123.3,121.9,121.5,119.1,118.5,111.8,108.4,52.7,48.5,23.3,22.6,16.3.ESI-MS m/z 471[M+H]⁺,HR-EI-MS:m/z 471.1743[M+H]⁺,(calcd forC₂₈H₂₇N₂O₃S⁺471.1737). methyl ester product (107mg,0.22mmol,1eq) was dissolved in 10mL of methanol, NaOH (40mg,1mmol,5eq) was added and refluxed for 4h, 2mL of HCl (1N) was added after evaporation to dryness and precipitated, filtered and recrystallized with methanol to obtain the product. Methyl- (E) -4- (3- (1- (5-ethylthiophen-2-yl) -1,3,4, 9-tetrahydro-2H-pyridine [3, 4-b)]Indol-2-yl) -3-oxo-1-en-1-yl) benzoic acid (compound VI-1). Yellow crystals, mp.175-177 ℃.¹H NMR(400MHz,DMSO-d₆)δ13.07(s,1H),11.10(s,1H),7.97(d,J＝8.2Hz,2H),7.91(d,J＝8.0Hz,2H),7.66(d,J＝15.1Hz,1H),7.56–7.43(m,2H),7.32(d,J＝8.0Hz,1H),7.10(t,J＝7.5Hz,1H),7.05–6.94(m,2H),6.69(d,J＝3.5Hz,1H),6.66(d,J＝3.5Hz,1H),4.68–4.38(m,1H),3.55–3.31(m,3H),2.88(d,J＝4.9Hz,2H),2.74(p,J＝9.1,8.3Hz,3H),1.19(t,J＝7.4Hz,3H).¹³C NMR(101MHz,DMSO)δ167.4,164.8,147.5,141.7,140.6,139.7,136.6,132.2,131.8,130.1,128.7,127.1,126.5,123.3,121.9,121.2,119.1,118.5,111.8,108.4,48.5,23.3,22.6,16.3.ESI-MS m/z 457[M+H]⁺,HR-EI-MS:m/z 457.1580[M+H]⁺,(calcd for C₂₇H₂₅N₂O₃S⁺,457.1580).

Compound and structure confirmation

Example 7Biological Activity assay

The compounds of the present application or optically pure enantiomers or solvates or hydrates thereof or pharmaceutically acceptable salts thereof or pharmaceutical compositions or formulations comprising the foregoing are useful for treating diseases or conditions associated with PDE5 in a mammal. By means of a corresponding administration, the use can improve local or systemic vascular conditions, in particular local or systemic arterial blood flow, and can therefore be used for the therapeutic or prophylactic treatment of Male Erectile Dysfunction (MED), Female Sexual Dysfunction (FSD), premature labour, dysmenorrhea, Benign Prostatic Hyperplasia (BPH), bladder outlet obstruction, incontinence, stable, unstable and variant (pregnanmer-toler) angina, hypertension, pulmonary Hypertension (HP), alzheimer's disease (senile dementia, AD), congestive heart failure, arteriosclerosis, stroke, peripheral vascular disease, reduced vascular patency, chronic asthma, bronchitis, allergic asthma, allergic rhinitis, glaucoma or diseases characterized by intestinal motility disorders. Preferred therapeutic conditions are Male Erectile Dysfunction (MED) and Pulmonary Arterial Hypertension (PAH).

1. In vitro inhibitory Activity on PDE5

Hereinafter, the present inventors determined the in vitro inhibitory activity of a compound (100nm) against PDE5 according to the method of J.biomol.Screen.,2002,7,215-222, wherein the inhibition of PDE5 is considered beneficial, wherein the inhibition of PDE5 is shown in tables 1,3 and 4, and the inhibition of PDE isoforms (PDE1-4, 6 and 11) is shown in Table 2, and the data below are the average of at least 2 experiments.

TABLE 1 (unit nm)

TABLE 2 (units nM)

TABLE 3

TABLE 4

The assay for inhibitory activity against PED-5 is further illustrated by compound II-2 as follows:

a45. mu.L enzymatic reaction system was prepared, with the following concentrations of the components: 40mM MOPS, pH 7.5,0.5mM EDTA，15mM MgCl₂0.15mg/ml BSA, 1mM DTT, 0.05% Proclin 200, 15ng/ml PDE5, 100nM FAM-Cyclic-3 ',5' -GMP and 100nM Compound II-2 (or an equivalent concentration of DMSO). After the reaction system was maintained at 25 ℃ for 1h, 100. mu.L of diluted IMAP solution was added and shaken at low speed at 25 ℃ for 1 h. The fluorescence values of the samples under excitation light at 360nm and 480nm were read and the experiment was repeated 3 times. Calculating the enzyme activity of PDE5 and the inhibition rate of compound II-2 on PDE 5. The results of the experiments on the inhibition of PED-5 by compound II-2 at a concentration of 100nM are shown in FIG. 1, with Tadalafil (Tadalafil) as a positive control. Among them, compound II-2 had an IC50 of 2.8nM and Tadalafil had an IC50 of 2.8 nM.

Experiments prove that the compound shown in the formula (I), the formula (II), the formula (V) and the formula (VI) have better inhibitory activity on PED-5, and the compound shown in the formula (I), the formula (II) and the formula (V) have better selectivity; the compounds of formula (IV) and formula (VII) have certain inhibitory effect on PED-5.

2. Measurement of diastolic Activity of NE-Pre-contracted mesenteric arteriole

The compound of the present invention has an effect of dilating blood vessels, particularly arterioles. The present invention is described in Shahid, M.D., Buys, E.S. assembling Murine Resistance array function Using Pressure physiology.J.Vis.Exp. (76), e50328, doi:10.3791/50328(2013), by isolating 2-3 grade mesenteric arterioles from rats and determining the effect of the compounds of the invention on the diameter of Norepinephrine (NE) pre-contracted mesenteric arterioles using DMT-112PP microvascular Pressure diameter perfusion system. Calculating the relaxation rate of the preferred compound on the vascular ring and obtaining the EC of the corresponding compound on the relaxation of the isolated arterioles₅₀The value is obtained. Preferred Compounds have diastolic Effect on isolated mesenteric arterioles in rats and their EC₅₀The values are shown in Table 5.

TABLE 5

Compound (I)	I-3	I-6	I-9	I-13	II-1	II-2	II-5	II-6	II-8	Tadalafil
											EC₅₀(nM)	725	71	30	55	191	63	225	58	103	78

Method for measuring the vasodilating activity of NE-preshrunk mesenteric arteriole vessels: an isolated vascular ring of mesenteric arterioles is prepared. A Wistar rat of 250-. Dissecting abdominal cavity, rapidly taking out intestinal tissuePre-cooled at 4 deg.C and charged with 95% O₂And 5% CO₂In the PSS solution of (1). The selected intestinal tissues are fixed in a culture dish by a pin. Parallel arteries and veins can be clearly seen by observation under a stereomicroscope, and the outsides of the vessels are wrapped by fat tissues. Wherein, the wall of the artery is thicker, and the branch point of the blood vessel is V-shaped; the vein wall is thin, and the branch point of the blood vessel is U-shaped. 2-3 grade mesenteric arterioles (the pipe diameter is about 110-.

The pressure, gravity and diameter levels of the DMT-112PP micro-vascular pressure diameter perfusion system are corrected in advance before the experiment. Firstly, one side of a prepared blood vessel ring is fixed at the liquid inlet end of a glass electrode in a system bath tank, and is fastened by a preset nylon knot, a certain pressure is applied to the liquid inlet end to flush out residues in the blood vessel, and then the blood vessel is fixed at the other end of the electrode and is fastened. After the blood vessel is fixed, the hollow glass electrodes at two ends are pressurized and perfused to simulate the physiological state of the blood vessel. The vessels were given a pre-perfusion pressure (10mmHg) and the perfusion pressure was raised to 50mmHg at a rate of 10mmHg per 5 min. During the equilibration process, the bath temperature was kept constant at 37 ℃ and the solution was changed every 20 min. After the blood vessels are balanced, the high potassium PSS solution and the acetylcholine are added in sequence to stimulate the blood vessels. The endothelial intact vascular ring with the K-PSS shrinkage rate of more than 80% and the Ach (10 mu M) relaxation rate of more than 60% is selected for experiments.

Addition of 20 μ M norepinephrine to pre-constrict vessels, concentration accumulation after vasoconstriction stabilized (10%^-11To 10^- ⁵M) adding a PDE5 inhibitor and recording the vascular diameter values at different concentrations of the compound, and administering the next concentration of drug after the vascular diameter indication for each concentration of drug has stabilized. After dosing was completed, the vessels were flushed with calcium-free PSS solution and the maximum diastolic vessel diameter at this time was recorded. The vasodilation rate was calculated for evaluation of the vasodilation effect of the compounds on blood vessels. The compound of the invention has a vasodilatation effect on blood vessels, wherein the vasodilatation effect of the compounds I-9, II-2 and I-13 on the concentration dependence of arteriole is shown in figure 2, and the effects of I-9 and II-2, I-13 increase with cumulative dose (10)^-11To 10^-5M) vasodilatation of blood vessels, and the result shows that the compound can improve the local arterial blood vessel flow and reduce the blood vessel pressure.

Claims

1. A compound, or a pharmaceutically acceptable salt thereof, having a structure represented by formula (X):

wherein X is selected from sulfur and oxygen;

R₁is a substituent on the five-membered heterocyclic ring, R₁The substituent can be one or more, and each substituent is independently selected from hydrogen, bromine and C₁-C₄The phenyl group forms benzofuran-2-yl or-5-methoxybenzofuran-2-yl with a five-membered heterocyclic fused ring in which the phenyl group is located;

the A group is fused to the five-membered ring of the indole with a common ring edge, the A group being selected from:

R₂is selected from C₂-C₄Alkyl groups of (a); r₃Selected from bipyridyl, 4- (3, 4-dimethoxyphenyl) pyrimidinyl; or-NH-ring closure to form the structure:

R₄selected from carboxyl groups.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, having a structure according to formula (I), (II), (IV), (V), (VI), (VII):

wherein the content of the first and second substances,

represents a single bond or a double bond when

When the compound is a double bond, the compound of the formula (II) is a double bond product after dehydrogenation at the C-11, C-11a position, and the structure of the compound is

R₁The number of the substituent groups is 1 or 2, and the substitution position is selected from C-3, C-4 and C-5;

R₁selected from hydrogen, bromine, C₁-C₃Alkyl and phenyl groups of (a); the phenyl and a five-membered heterocyclic fused ring in which the phenyl is positioned form benzofuran-2-yl or-5-methoxybenzofuran-2-yl;

R₂selected from ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl;

R₃selected from bipyridyl, 4- (3, 4-dimethoxyphenyl) pyrimidinyl; or at

In, R₃and-NH-ring to form the structure:

3. the compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein in the compound of formula (I), X is selected from O and S; r₁Selected from hydrogen, bromine, methyl and ethyl;

in the compound of formula (I), R₁The substitution position(s) is selected from the C-3 and C-5 positions;

in the compound of formula (II), X is selected from O and S; r₁Selected from methyl and ethyl; r₂Is selected from C₂-C₄Alkyl groups of (a);

in the compound of formula (IV), X is selected from O and S; r₁1 or 2 substituents R₁The substituents being selected from C₁-C₃The phenyl group forms benzofuran-2-yl or-5-methoxybenzofuran-2-yl with a five-membered heterocyclic fused ring in which the phenyl group is located;

R₃selected from bipyridyl, 4- (3, 4-dimethoxyphenyl) pyrimidinyl, or R₃with-NH-ring to form a radical selected from

A group of (a);

in the compound of formula (IV), R₁The substitution position(s) is selected from the C-3, C-4 and C-5 positions.

4. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein in the compound of formula (II), R₂Selected from ethyl, propyl, isopropyl, n-butyl, isobutyl and tert-butyl.

5. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein in the compound of formula (IV), X is O or S; r₁The substituent is 1, R₁Phenyl and five-membered heterocyclic fused ring in which the phenyl is positioned form benzofuran-2-yl or-5 methoxy benzofuran-2-yl.

6. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein in the compound of formula (IV), X is O or S; r₁The substituent is 1, R₁Is methyl, ethyl or n-propyl, R₁The substitution position of (A) is C-5 or C-3.

7. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein in the compound of formula (IV), X is O or S; r₁When the number of the substituent is 2, the substituent is methyl, and the substitution positions are C-4 position and C-5 position respectively.

8. As in claimThe compound of claim 2, wherein X is S and R is R in the compound of formula (IV), or a pharmaceutically acceptable salt thereof₁The substituent is 1, R₁The substituents being methyl, ethyl or n-propyl, R₁The substitution position of (A) is C-5 or C-3, R₃By ring closure with-NH-groups

9. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, selected from the following compounds:

2- (5- (3, 4-dimethoxyphenyl) pyrimidin-2-yl) -1- (5-ethylthiophen-2-yl) -1,2,3, 9-tetrahydro-4H-pyridin [3,4-b ] indol-4-one.

10. A composition or formulation comprising a compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof;

the preparation comprises tablet, capsule, lozenge, sugar-coated pill, granule, powder, solution, emulsion, suspension, dispersion, syrup, gel, and aerosol;

the composition or preparation comprises a pharmaceutical composition or preparation for human use and a pharmaceutical composition or preparation for veterinary use.

11. Use of a compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or formulation of claim 10, for the manufacture of a medicament for treating, ameliorating or preventing a disease or condition associated with PDE5 in a mammal, wherein the disease or condition is selected from, but is not limited to, male erectile dysfunction, male testicular ischemia, female sexual dysfunction, premature labor, dysmenorrhea, benign prostatic hyperplasia, bladder outlet obstruction, incontinence, stable, unstable and variant angina, hypertension, pulmonary hypertension, alzheimer's disease, congestive heart failure, arteriosclerosis, stroke, peripheral vascular disease, reduced vascular patency disorders, chronic asthma, bronchitis, allergic asthma, allergic rhinitis, glaucoma, or a disorder characterized by intestinal motility disorders.

12. Use of a compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or formulation of claim 10, in the manufacture of a medicament for treating, ameliorating or preventing a vascular-related disease or condition in a mammal, wherein the disease or condition is selected from, but is not limited to, male erectile dysfunction, male testicular ischemia, female sexual dysfunction, premature labor, dysmenorrhea, benign prostatic hyperplasia, bladder outlet obstruction, incontinence, stable, unstable and variant angina, hypertension, pulmonary hypertension, alzheimer's disease, congestive heart failure, arteriosclerosis, stroke, peripheral vascular disease, reduced vascular patency disorders, chronic asthma, bronchitis, allergic asthma, allergic rhinitis, glaucoma, or a disorder characterized by intestinal motility disorders.