CN105384738A - Heterocyclic compound as protein kinase inhibitor and preparation method and application thereof - Google Patents

Abstract

The present invention relates to compounds of formula (I) and (II) and pharmaceutically acceptable salts, prodrugs and solvates thereof, wherein R ₁ , R ₂ , R ₃ , A and X are as defined in the description. Such compounds are protein kinase inhibitors, especially protein kinase inhibitor Mek inhibitors, and are useful in the treatment of cancer and inflammation in mammals. The invention also discloses the preparation method of the compound of formula (I) and (II) and the pharmaceutical composition containing the compound.

Description

Heterocyclic compound as protein kinase inhibitor and its preparation method and use

technical field

The present invention relates to protein kinase (especially protein kinase Mek) inhibitors, more specifically, to imidazopyridine compounds as protein kinase (especially protein kinase Mek) inhibitors and pharmaceutically acceptable salts, prodrugs, Solvates, and compositions comprising these substances, and relate to the preparation method of said imidazopyridine compounds, also relate to said imidazopyridine derivatives and pharmaceutically acceptable salts, prodrugs and solvates thereof as protein kinases ( In particular the use of protein kinase Mek) inhibitors.

Background technique

Cellular signal transduction controlled by growth factors and protein kinases plays an important role in cell growth, proliferation and differentiation. In the growth of normal cells, growth factors (such as PDGF or EGF, etc.) activate the MAP (Mitogen-activatingprotein) kinase signal transduction channel through receptor activation (such as ErbB2, EGFR, PDGFR, etc.). The Ras/Raf/Mek/Erk signaling mechanism is one of the most important pathways for cell growth. In proliferative diseases, genetic mutations or overexpression of growth factor receptors or their downstream protein kinases lead to uncontrolled cell growth and ultimately cancer. For example, in some cancers, due to gene mutations, the signal transduction mechanism is continuously activated, which leads to the continuous production of some growth factors, and finally leads to the loss of control of cell growth, resulting in cancer. Statistics show that 50% of colon cancer and more than 90% of pancreatic cancer are caused by Ras gene mutation; more than 60% of malignant melanoma are caused by bRaf gene mutation. Studies have shown that the Ras/Raf/Mek/Erk signaling mechanism is continuously activated or overactivated in a variety of cancers, such as pancreatic cancer, colon cancer, lung cancer, bladder cancer, kidney cancer, skin cancer, breast cancer, etc. Wait.

Since hyperactivation of this signaling mechanism plays an important role in the proliferation and differentiation of cancer cells, inhibition of this pathway could be useful in the treatment of such hyperproliferative diseases. Mek is located at the downstream target of Ras and Raf and plays a key role in this pathway. The substrate of Mek phosphorylation is MAP kinase Erk. If Mek is inhibited, the Ras/Raf/Mek/Erk signaling pathway will be turned off, thereby inhibiting the proliferation of cancer cells. Therefore, Mek inhibitors can inhibit the growth of cancer cells, especially for cancers caused by overactivation of Ras or Raf. At the same time Mek is also involved in inflammatory diseases and symptoms, including acute and chronic inflammation.

Mek inhibitors have shown certain efficacy in pharmacodynamic experiments in nude mice. Recently, some Mek inhibitors have entered the clinic and have also shown certain efficacy. Therefore, Mek is a potential druggable new target, and because of this, more and more Mek inhibitors are being developed and reported. For example, WO98/43960; WO99/01421; WO99/01426; WO00/41505; WO00/42002; WO00/41003; WO00/41994; WO02/06213; WO03/077914; WO03/077855; WO03/077914; WO05/023251; WO05/023759; 009975; WO05/046665; WO06/134469; WO07/044084; WO07/014011; WO07/121269; WO07/121481; WO08/120004; WO08/124085; WO08/125820; WO09/018238; WO09/074827; WO09/013426; WO2007/044084 and the like.

Contents of the invention

One aspect of the present invention provides compounds of formula (I) and (II) and pharmaceutically acceptable salts, prodrugs and solvates thereof

in

R is selected from hydrogen, halogen, cyano, nitro, azido, hydroxyl, C ¹ -C ₁₀ alkoxy, halogenated C ₁ -C ₁₀ alkoxy, mercapto, C ₁ -C _{10 alkylthio} , halogenated C ₁ -C ₁₀ alkylthio, carboxyl, -OC(O)H, amino, C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ - C ₁₀ cycloalkyl;

R ² is selected from hydrogen, halogen, nitro, azido, hydroxyl, C ₁ -C ₁₀ alkyl, C ₁ -C ₁₀ alkoxy, halogenated C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ Alkylthio, halogenated C ₁ -C ₁₀ alkylthio, carboxyl, -OC(O)H, amino, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl ;

R ₃ is selected from hydrogen, C ₁ -C ₁₂ alkyl, C ₂ -C ₁₂ alkenyl, C ₂ -C ₁₂ alkynyl, C ₃ -C ₁₂ cycloalkyl, C ₃ -C ₁₂ cycloalkyl C ₁ - C ₁₂ alkyl, C ₁ -C ₁₂ alkyl, C ₃ -C ₁₂ cycloalkyl, the alkyl, alkenyl, alkynyl and cycloalkyl can each be independently replaced by one to three groups selected from the following groups Optional substitution: -ORa, -NRaRb, -NRaC(O)Rb, heterocycloalkyl and heterocyclic aryl, where the heterocycloalkyl and heteroaryl can be substituted by one to three selected from the following Arbitrary substitution of groups: C ₁ -C ₅ alkyl, -ORa, -NRaRb;

X represents formula (i):

in

Y is selected from O, -ONR ₈ , -NR ₈ O, -NR ₈ C(O), -NR ₈ SO ₂ ;

Z is selected from hydrogen, C ₁ -C ₅ methylene chains, said methylene can be optionally substituted by one to three W';

R ₈ is selected from hydrogen, C ₁ -C ₁₂ alkyl, -COR ₉ , and the alkyl can be optionally substituted by halogen, -ORa, -NRaRb;

R ₉ is selected from hydrogen, C ₁ -C ₁₂ alkyl, said alkyl can be optionally substituted by halogen, -ORa, -NRaRb;

or X represents formula (ii):

in

Y ₁ and Y ₂ may be the same or different, and they each independently represent a single bond, -CO-, -COO-, -O-, -OCO-, -NRa-, -SO ₂ -;

Z' represents a C ₁ -C ₁₀ chain hydrocarbon group, and the hydrocarbon group can be optionally substituted by one to three W';

In the above formula (i) and formula (ii),

W and W' can be the same or different, and they are each independently selected from halogen, C ₁ -C ₅ alkyl, -O-, -ORa, -COORa, -COOCORa, -CO-halogen, -OCORa, -CONRaRb, -SRa, -SORa, -SO ₂ Ra, -NRaRb, -NRaCORb, -NRaSO ₂ Rb, -SO ₂ NRaRb, heterocycloalkyl or heteroaryl, the heterocycloalkyl and heteroaryl can be One to three substituents selected from the following substituents are optionally substituted: C ₁ -C ₅ alkyl, -ORa, -NRaRb; the alkyl can be optionally substituted by hydroxyl, C ₁ -C ₅ alkoxy or amino;

The above-mentioned groups can be connected to each other except oxygen and halogen to form cycloalkyl or heterocycloalkyl, and the cycloalkyl or heterocycloalkyl can be arbitrarily substituted by substituents selected from the following groups: -ORa, -NRaRb, by -ORa optionally substituted C ₁ -C ₅ alkyl;

Ra and Rb may be the same or different, and they are each independently selected from hydrogen, C ₁ -C ₅ alkyl, and the alkyl may be optionally substituted by one to three substituents selected from: hydroxyl, C ₁ -C ₅ alkoxy, amino.

When X represents formula (i), the compounds of formulas (I) and (II) have the following structures:

When X represents formula (ii), the compounds of formulas (I) and (II) have the structure of the following formula (I-2):

When Y represents -O-, and W represents OH, the compound of formula (I-1) has the structure of the following formula (I-1-a):

When Y represents -NHO-, the compound of formula (I-1) has the following structure:

When Y represents -NR ₈ O-, the compound of formula (I-1) has the structure of the following formula (I-1-c):

When Y represents _a single bond and Y represents -O-, the compound of formula (I- ₂ ) has the structure of the following formula (I-2-A):

Another aspect of the present invention provides the preparation method of the compound of formula (I):

(A) Route 1

(B) Route 2

(C) The compound of formula (I-1-a) is prepared according to the following route 3:

Wherein, the synthetic reference route 2 of compound 11

(D) The compound of formula (I-1-b) is prepared according to the following route 4:

Wherein, the synthetic reference route 2 of compound 11

(E) The compound of formula (I-1-c) is prepared according to the following route 5:

(F) The compound of formula (I-2-A) is prepared according to the following route 6:

(G) The compound of formula (I-2-A) is prepared according to the following route 7:

Yet another aspect of the present invention provides pharmaceutical compositions comprising compounds of formula (I) and (II) or pharmaceutically acceptable salts, prodrugs and solvates thereof.

Another aspect of the present invention provides formula (I) and (II) compound or its pharmaceutically acceptable salt, prodrug and solvate are used for the manufacture of treatment mammalian tumor, acute and chronic inflammatory disease, inflammatory bowel disease , skin diseases, diabetes, eye diseases, diseases associated with angiogenesis or angiogenesis in mammals, diseases associated with chronic pain, and other diseases modulated by the Mek cascade.

detailed description

If not otherwise indicated, the entire disclosure content of the present invention adopts the following term definitions:

The term "prodrug" refers to any derivative that can be converted into the corresponding active drug compound in vivo. In one embodiment, when a compound of the invention contains a hydroxyl group, its prodrug may be its ester with a suitable acid, including, for example, lactic acid, citric acid, ascorbic acid, and the like.

The term "pharmaceutically acceptable salt", unless otherwise stated, includes salts of acidic groups that may be present in the compounds of the present invention (such as, but not limited to, potassium salts, sodium salts, magnesium salts, calcium salts, etc.) or Salts of basic groups (eg, but not limited to, sulfates, hydrochlorides, phosphates, nitrates, carbonates, etc.).

The term "solvate" refers to a complex molecular compound formed by solute molecules or ions attracting adjacent solvent molecules through intermolecular forces such as Coulomb force, van der Waals force, charge transfer force, and hydrogen bond in a solution. In one embodiment, the solvent is water, ie the compounds of the invention form hydrates.

The alkyl, alkenyl, alkynyl, cycloalkyl moieties may each independently be optionally substituted by one or more groups selected from the group consisting of hydroxyl, oxo, halogen, cyano, nitro, trifluoromethyl group, azido group, amino group, carboxyl group, mercapto group.

Saturated or unsaturated hydrocarbon radicals, such as alkyl, alkanediyl or alkenyl, including in combination with heteroatoms, such as alkoxy, can each be straight-chain or branched.

Depending on the substituents, the compounds of the formulas (I) and (II) can exist as optical isomers or isomer mixtures of different composition, which mixtures can, if appropriate, be separated by conventional means. The present invention provides pure isomers and isomer mixtures, methods for their preparation and use, and compositions comprising them. For the sake of simplicity, they are referred to below as compounds of the formulas (I) and (II), referring both to the pure optical isomers and, if appropriate, to mixtures of isomers in varying proportions.

In some embodiments of the present invention, compounds of formula (I) and (II) and pharmaceutically acceptable salts, prodrugs and solvates thereof are provided

in

A is selected from C and N;

R is selected from hydrogen, halogen, cyano, nitro, azido, hydroxyl, C ¹ -C ₁₀ alkoxy, halogenated C ₁ -C ₁₀ alkoxy, mercapto, C ₁ -C _{10 alkylthio} , halogenated C ₁ -C ₁₀ alkylthio, carboxyl, -OC(O)H, amino, C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ - C ₁₀ cycloalkyl;

R ² is selected from hydrogen, halogen, nitro, azido, hydroxyl, C ₁ -C ₁₀ alkyl, C ₁ -C ₁₀ alkoxy, halogenated C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ Alkylthio, halogenated C ₁ -C ₁₀ alkylthio, carboxyl, -OC(O)H, amino, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl ;

R ₃ is selected from hydrogen, C ₁ -C ₁₂ alkyl, C ₂ -C ₁₂ alkenyl, C ₂ -C ₁₂ alkynyl, C ₃ -C ₁₂ cycloalkyl, C ₃ -C ₁₂ cycloalkyl C ₁ - C ₁₂ alkyl, C ₁ -C ₁₂ alkyl, C ₃ -C ₁₂ cycloalkyl, the alkyl, alkenyl, alkynyl and cycloalkyl can each be independently replaced by one to three groups selected from the following groups Optional substitution: -ORa, -NRaRb, -NRaC(O)Rb, heterocycloalkyl and heterocyclic aryl, where the heterocycloalkyl and heteroaryl can be substituted by one to three selected from the following Arbitrary substitution of groups: C ₁ -C ₅ alkyl, -ORa, -NRaRb;

X represents formula (i):

in

Y is selected from O, -ONR ₈ , -NR ₈ O, -NR ₈ C(O), -NR ₈ SO ₂ ;

Z is selected from hydrogen, C ₁ -C ₅ methylene chains, said methylene can be optionally substituted by one to three W';

R ₈ is selected from hydrogen, C ₁ -C ₁₂ alkyl, -COR ₉ , and the alkyl can be optionally substituted by halogen, -ORa, -NRaRb;

R ₉ is selected from hydrogen, C ₁ -C ₁₂ alkyl, said alkyl can be optionally substituted by halogen, -ORa, -NRaRb;

or X represents formula (ii):

in

Y ₁ and Y ₂ may be the same or different, and they each independently represent a single bond, -CO-, -COO-, -O-, -OCO-, -NRa-, -SO ₂ -;

Z' represents a C ₁ -C ₁₀ chain hydrocarbon group, and the hydrocarbon group can be optionally substituted by one to three W';

In the above formula (i) and formula (ii),

W and W' can be the same or different, and they are each independently selected from halogen, C ₁ -C ₅ alkyl, -O-, -ORa, -COORa, -COOCORa, -CO-halogen, -OCORa, -CONRaRb, -SRa, -SORa, -SO ₂ Ra, -NRaRb, -NRaCORb, -NRaSO ₂ Rb, -SO ₂ NRaRb, heterocycloalkyl or heteroaryl, the heterocycloalkyl and heteroaryl can be One to three substituents selected from the following substituents are optionally substituted: C ₁ -C ₅ alkyl, -ORa, -NRaRb; the alkyl can be optionally substituted by hydroxyl, C ₁ -C ₅ alkoxy or amino;

The above-mentioned groups can be connected to each other except oxygen and halogen to form cycloalkyl or heterocycloalkyl, and the cycloalkyl or heterocycloalkyl can be arbitrarily substituted by substituents selected from the following groups: -ORa, -NRaRb, by -ORa optionally substituted C ₁ -C ₅ alkyl;

Ra and Rb may be the same or different, and they are each independently selected from hydrogen, C ₁ -C ₅ alkyl, and the alkyl may be optionally substituted by one to three substituents selected from: hydroxyl, C ₁ -C ₅ alkoxy, amino.

Above-mentioned general formula compound (I) and (II) and following preferred formula (I) and (II) compound preferably following substituent or group:

R ₁ is selected from fluorine, chlorine, bromine, iodine, C ₁ -C ₄ alkoxy, C ₁ -C ₄ alkylthio, halo-C ₁ -C ₄ alkoxy, halo-C ₁ -C ₄ Alkylthio, C ₁ -C ₄ alkyl, halogenated C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl;

R ₂ is preferably hydrogen, halogen or C ₁ -C ₄ alkyl;

R ₃ is preferably C ₁ -C ₄ alkyl, C ₃ -C ₄ cycloalkyl, C ₃ -C ₄ cycloalkyl C ₁ -C ₄ alkyl or C ₁ unsubstituted or substituted by 1 to 6 hydroxyl groups -C ₄ alkylC ₃ -C ₄ cycloalkyl.

R ₁ is selected from bromine, iodine, C ₁ -C ₂ alkylthio, halo-C ₁ -C ₂ alkylthio, C ₁ -C ₂ alkoxy, halo-C ₁ -C ₂ alkoxy, C ₁ -C ₂ alkyl, halogenated C ₁ -C ₂ alkyl, C ₂ -C ₃ alkenyl, C ₂ -C ₃ alkynyl;

R ₂ is more preferably hydrogen, fluorine, chlorine, bromine or C ₁ -C ₂ alkyl;

R ₃ is more preferably unsubstituted or C ₁ -C ₃ alkyl, C ₃ -C ₄ cycloalkyl, C ₃ -C ₄ cycloalkyl C ₁ -C ₃ alkyl or C substituted by 1 to 3 hydroxyl groups ₁ -C ₃ alkylC ₃ -C ₄ cycloalkyl.

R is selected from _bromine , iodine, methylthio, trifluoromethylthio, methoxy, trifluoromethoxy, methyl, trifluoromethyl, vinyl, ethynyl;

R is especially preferably fluorine, chlorine or methyl _;

R is especially preferably 2-hydroxyethyl, ₃ -hydroxyethyl, 2,3-dihydroxypropyl, 1-hydroxymethyl-2-hydroxyethyl, 2-methyl-3-hydroxypropyl, cyclopropyl , cyclopropylmethyl or 1-(2,3-hydroxypropyl)cyclopropyl.

When X is preferably -Y-Z-W represented by formula (i),

Y is preferably -O-, -NR ₈ O-, -NR ₈ C(O)-, -NR ₈ SO ₂ -;

Y is more preferably -O-, -NR ₈ O-, -NR ₈ C(O)-;

Y is especially preferably -NR ₈ O-, -NR ₈ C(O)-;

R ₈ is preferably hydrogen, C ₁ -C ₆ alkyl, -COR ₉ ;

R ₈ is more preferably hydrogen, C ₁ -C ₃ alkyl, -COR ₉ ;

R ₈ is especially preferably hydrogen, methyl, -COH, -COCH ₃ , -COCH ₂ CH ₃ , -COCH(CH ₃ ) ₂ , -COCH ₂ OH;

Z is preferably hydrogen, a C ₁ -C ₅ methylene chain, and the methylene group may be optionally substituted by one to three W';

Z is more preferably hydrogen, -CH ₂ -, -(CH ₂ ) ₂ -, -(CH ₂ ) ₃ -, -CH ₂ (CH ₃ )CH ₂ -, -CH ₂ C(CH ₃ ) ₂ -, -CH ₂ C(CH ₃ ) ₂ CH ₂ -, -CH ₂ C(CH ₂ CH ₂ )CH ₂ -, and can be optionally substituted by W';

W and W' can be the same or different, and they are independently preferably halogen, C ₁ -C ₅ alkyl, -ORa, -COORa, -COOCORa, -OCORa, -CONRaRb, -SRa, -SORa, -SO ₂ Ra , -NRaRb, -NRaCORb, -NRaSO ₂ Rb, -SO ₂ NRaRb, heterocycloalkyl or heteroaryl;

W and W' can be the same or different, and they are independently more preferably halogen, C ₁ -C ₃ alkyl, -ORa, -COORa, -COOCORa, -OCORa, -CONRaRb, -SRa, -SORa, -SO ₂ Ra, -NRaRb, -NRaCORb, -SO ₂ NRaRb, heterocycloalkyl or heteroaryl;

W and W' may be the same or different, and they are each independently preferably hydrogen, hydroxyl, methyl, -O(CH ₂ ) ₂ OH, -C(O)OCH ₃ , -CONHCH ₃ , -CONHCH ₂ CH ₃ , -CONHCH(CH ₃ ) ₂ , -CON(CH ₃ ) ₂ , -NHCH ₃ , -NH(CH ₂ ) ₂ OH, -NHCOCH ₃ , -NHCOCH ₂ CH ₃ , -NHCOCH(CH ₃ ) ₂ , -SCH ₃ , -SOCH ₃ , 1H-imidazol-2-yl, morpholin-4-yl, 4-hydroxypiperidinyl.

When X is a substituent represented by formula (ii),

X preferred

Z' is preferably a C ₁ -C ₃ chain hydrocarbon group, which can be optionally substituted by one to three W's;

W' is preferably C ₁ -C ₃ alkyl, -ORa, -COORa, -COOCORa, -CO-halogen, -OCORa, -CONRaRb, -SRa, -SORa, -SO ₂ Ra, -NRaRb, -NRaCORb, -NRaSO ₂ Rb, -SO ₂ NRaRb;

Ra and Rb can be the same or different, and they are each independently selected from hydrogen, C ₁ -C ₃ alkyl, and the alkyl can be optionally substituted by one to three substituents selected from: hydroxyl, C ₁ -C ₃ alkoxy, amino.

X is more preferred

Z' is more preferably -CH ₂ -, -(CH ₂ ) ₂ -, -(CH ₂ ) ₃ -, -(CH ₂ ) ₄ -, wherein any hydrogen atom is optionally substituted by one to three W';

W' is more preferably hydroxyl, C ₁ -C ₂ alkyl.

X is especially preferred

Groups in the above-mentioned compounds of general formula (I) and (II) and preferred compounds of formula (I) and (II) can be combined with each other, that is, including non-preferred groups in the compounds of general formula (I) and (II) , and combinations between substituents and groups of different preferred levels. The various combinations above apply both to the final product and thus also to the precursors and intermediates.

Compounds of formula (I) and (II) comprising the abovementioned preferred substituents and groups and combinations thereof are preferred according to the invention.

More preferred in the present invention are compounds of formula (I) and (II) comprising the more preferred substituents and groups described above and combinations thereof.

Particularly preferred according to the invention are compounds of the formulas (I) and (II) which comprise the abovementioned particularly preferred substituents and groups and combinations thereof.

Especially preferred according to the invention are compounds of formula (I) and (II) which comprise the abovementioned particularly preferred substituents and groups and combinations thereof.

Saturated or unsaturated hydrocarbon radicals, such as C ₁ -C ₁₀ -alkyl, alkanediyl or alkenyl, including in combination with heteroatoms, eg alkoxy, can each be straight-chain or branched.

Unless stated otherwise, optionally substituted radicals can be monosubstituted or polysubstituted, where in the case of polysubstitution the substituents can be identical or different.

In some preferred embodiments, the following compounds may be mentioned:

In a more preferred embodiment, the following compounds may be mentioned:

synthesis

Suitable solvents commonly used in organic reactions can be used in the following reactions in each step of the preparation method of the present invention, such as, but not limited to, aliphatic and aromatic, optionally hydrocarbons or halogenated hydrocarbons (such as pentane, hexane, heptane, alkane, cyclohexane, petroleum ether, gasoline, volatile oil, benzene, toluene, xylene, methylene chloride, dichloroethane, chloroform, carbon tetrachloride, chlorobenzene and o-dichlorobenzene), aliphatic and aromatic Optional alcohols (such as methanol, ethanol, propanol, isopropanol, tert-butanol, ethylene glycol, etc.), ethers (such as diethyl ether and dibutyl ether, ethylene glycol dimethyl ether and diethylene glycol di methyl ether, tetrahydrofuran and dioxane, etc.), esters (such as methyl acetate or ethyl acetate, etc.), nitriles (such as acetonitrile or propionitrile, etc.), ketones (such as acetone, butanone, etc.), amides (such as dimethyl formamide, dimethylacetamide and N-methylpyrrolidone, etc.), and dimethyl sulfoxide, tetramethylene sulfone and hexamethylphosphoric triamide and N,N-dimethylpropylene urea (DMPU )Wait.

use

The compound of the present invention can be used for treating following diseases, for example: tumor (tumor), for example: hemangioma (hemangioma), glioma (glioma), melanoma (melanoma), Kaposi's sarcoma (sarcoma) and ovarian cancer (ovariancancer), Breast cancer, lung cancer, pancreatic cancer, prostate cancer, colon cancer, breast cancer and other gastrointestinal cancers, etc.; chronic inflammatory diseases, such as Rheumatoid arthritis, diseases associated with vasculogenesis or angiogenesis in mammals; atherosclerosis, inflammatory bowel disease; skin diseases such as silver Psoriasis, excema, and sceroderma, diabetes, diabetic retinopathy, retinopathy of prematurity, age-ralated macular degeneration; disorders associated with chronic pain , including neuralgia and diseases modulated by the MEK cascade, such as postoperative pain, phantomlimb pain, burn pain, gout, trigeminal neuralgia, acute hepatic pain and posthepatic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma, vasculitis, crush injury, constriction injury, tissue damage (tissue injury), post-surgical pain, arthritis pain or limb amputation pain.

1. Dosage

A person skilled in the art can determine the dose for a patient according to known methods, taking into consideration factors such as age, body weight, health condition, the type of disease being treated and the presence of other drugs. Generally, the effective amount is 0.1 to 1000 mg/kg body weight per day, preferably 1 to 300 mg/kg body weight per day. For an adult subject of normal weight, the daily dose is usually 10 to 2500 mg. Commercially available 100 mg, 200 mg, 300 mg or 400 mg formulations can be administered according to published methods.

2. Preparations

The compounds of the present invention may be administered by any suitable route of administration, including systemic and topical administration. Systemic administration includes: oral, parenteral, transdermal, or rectal administration; or administration by inhalation. Parenteral administration means a route of administration other than enteral administration or transdermal administration, and is usually administered by injection or infusion. Parenteral administration includes intravenous administration, intramuscular administration, and subcutaneous injection or infusion. Topical administration includes application to the skin as well as intraocular, ophthalmic, intravaginal and intranasal administration.

Compounds of the present invention and one or more pharmaceutically acceptable excipients may be formulated into dosage forms suitable for administration to patients by the desired route of administration, such as tablets, capsules, pills, lozenges, Powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, and the like.

Suitable pharmaceutically acceptable excipients will vary according to the particular dosage form chosen. They include binders, lubricants, glidants, disintegrants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, flavoring agents, taste-masking agents, anti-caking agents, diluents Agents, chelating agents, plasticizers, viscosity modifiers, antioxidants, preservatives, stabilizers, surfactants, emulsifiers and buffers.

The pharmaceutical compositions of this invention are prepared using techniques and methods known to those skilled in the art.

Synthetic Example:

Example 1.5-(2-fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-8-((3-oxomorpholine)methyl)imidazo[1,5-a ] Synthesis of pyridine-6-carboxamide:

Step 1: Synthesis of 3,5-dichloro-6-methyl-2H-1,4-oxazin-2-one

According to European Journal of Organic Chemistry; nb, 30; (2007); p.4995-4998. Preparation of the title compound ¹ H-NMR (400MHz, DMSO-d ₆ ) δ 2.30 (s, 3H).

Step 2: Synthesis of ethyl 2,6-dichloro-5-methylnicotinate

The title compound was prepared according to synthesis; nb, 9; (1991); p.765-768.

Step 3: Synthesis of ethyl 6-chloro-2-(2-fluoro-4-iodophenylamino)-5-methylnicotinate

The compound 2-fluoro-4-iodoaniline (12.15g, 51.26mmol) was added to the reaction flask, anhydrous THF (50ml) was added, and after cooling to -78°C under a nitrogen atmosphere, LiHMDS was added dropwise to the reaction solution (1M, 128.5ml, 128.5mmol), after reacting at -78°C for 30 minutes, the compound 2,6-dichloro-5-methylnicotinic acid ethyl ester (10g, 42.72mmol) was added, and the dry ice was removed after the addition, Continue to react for 3 hours. After the reaction was completed, the reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain the product. (15g, yield: 81%) [M+H] ⁺ = 435.6

Step 4: Synthesis of ethyl 6-cyano-2-(2-fluoro-4-iodophenylamino)-5-methylnicotinate

The compound 6-chloro-2-(2-fluoro-4-iodophenylamino)-5-methylnicotinic acid ethyl ester (10.0g, 23.01mmol) was added to the reaction flask, DMF (50ml) was added, and then di Zinc cyanide (1.77g, 16.80mmol) and Pd(PPh ₃ ) ₄ (2.66g, 2.30mmol) were heated to 100°C for 8 hours under the protection of nitrogen and reacted for 8 hours. After the reaction was completed, it was diluted with ethyl acetate and filtered. The organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain the target product. (7.6g, yield: 77%).[M+H] ⁺ =426.2

Step 5: 6-aminomethyl-2-(2-fluoro-4-iodophenylamino)-5-methylnicotinic acid ethyl ester

Compound 6-cyano-2-(2-fluoro-4-iodophenylamino)-5-methylnicotinic acid ethyl ester (7.0g, 16.46mmol) was added to the reaction flask, methanol (80ml) was added, and then Cobalt chloride (4.28g, 32.93mmol), after stirring at room temperature for 10 minutes, sodium borohydride (6.23g, 164.63mmol) was added in batches under ice-water bath cooling, after the addition was completed, the reaction was continued for 15 minutes under ice-water bath, and then The reaction was continued for 1 hour at room temperature. Quench the reaction with concentrated hydrochloric acid, and stir at room temperature for 15 minutes, filter to remove the white solid, and wash with dichloromethane, concentrate the organic phase, and dissolve the obtained crude product in ethyl acetate, and wash with saturated sodium carbonate solution, water, and saturated salt Washed with water, dried over anhydrous sodium sulfate, concentrated to obtain the target product. (3.5g, Yield: 49.5%).[M+H] ⁺ =430.2 Step 6: 2-((2-fluoro-4-iodophenylamino)-6-(formylaminomethyl)-5- Synthesis of ethyl methylnicotinate

The compound 6-aminomethyl-2-(2-fluoro-4-iodophenylamino)-5-methylnicotinic acid ethyl ester (3.5g, 8.15mmol) was added to the reaction flask, formic acid (30ml) was added, and the After cooling to 0°C in an ice bath, add acetic anhydride (6ml), react at room temperature for 2 hours, concentrate the reaction solution, dissolve it in dichloromethane, wash with saturated sodium carbonate, wash with salt water, dry over anhydrous sodium sulfate, and concentrate to obtain the target product. (3.7g, yield: 99%)

Step 7: Synthesis of ethyl 5-(2-fluoro-4-iodophenylamino)-8-methylimidazo[1,5-a]pyridine-6-carboxylate

Add the compound 2-((2-fluoro-4-iodophenylamino)-6-(formylaminomethyl)-5-methylnicotinic acid ethyl ester (3.7g, 8.09mmol) into the reaction flask, add toluene (10ml), then add phosphorus oxychloride (2.48g, 16.18mmol), heat to 95 ℃ and react for 1 hour, cool to room temperature, the residue obtained after concentration is dissolved in ethyl acetate, and washed with saturated sodium carbonate solution, Wash with saturated brine, dry the organic phase with anhydrous sodium sulfate, concentrate, and separate by column chromatography to obtain the target product. (2.1g, yield: 59%).[M+H] ⁺ =440.2

Step 8: Synthesis of ethyl 8-(bromomethyl)-5-(2-fluoro-4-iodophenylamino)imidazo[1,5-a]pyridine-6-carboxylate

Add the compound 5-(2-fluoro-4-iodophenylamino)-8-methylimidazo[1,5-a]pyridine-6-carboxylic acid ethyl ester (2.10g, 4.78mmol) into the reaction flask, Carbon tetrachloride (20ml), NBS (0.94g, 5.26mmol) and benzoyl peroxide (BPO) (112mg, 0.47mmol) were then added. Heated to 60°C for 4 hours, cooled to room temperature, quenched with saturated sodium thiosulfate solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain the product. (1.8g, yield: 72%)

Step 9: 5-(2-Fluoro-4-iodophenylamino)-8-((4-(methylamino)-4-oxobutoxyamino)methyl)imidazo[1,5-a ] Synthesis of ethyl pyridine-6-carboxylate:

The compound 8-(bromomethyl)-5-(2-fluoro-4-iodophenylamino)imidazo[1,5-a]pyridine-6-carboxylic acid ethyl ester (150mg, 0.29mmol) was added to the reaction In the bottle, add acetonitrile (3ml), then add potassium carbonate (60mg, 0.43mmol) and 4-(aminooxy)-N-methylbutyramide (58mg, 0.43mmol), react at 60°C for 4 hours, cool Return to room temperature, add water to wash and extract with ethyl acetate, dry over anhydrous sodium sulfate, concentrate, and separate by column chromatography to obtain the product. (120 mg, yield: 73%). [M+H] ⁺ ＝570.4

Step 10: 5-(2-Fluoro-4-iodophenylamino)-8-((4-(methylamino)-4-oxobutoxyamino)methyl)-N-(2-(ethylene Synthesis of oxy)ethoxy)imidazo[1,5-a]pyridine-6-carboxamide

The compound O-(2-(vinyloxy)ethyl)hydroxylamine (33mg, 0.32mmol) was added to the reaction flask, anhydrous THF (1ml) was added, and after cooling to -78°C under a nitrogen atmosphere, the reaction LiHMDS (1M, 0.63ml, 0.63mmol) was added dropwise into the solution, and after reacting at -78°C for 30 minutes, the compound 5-(2-fluoro-4-iodophenylamino)-8-((4-(methyl Amino)-4-oxobutoxyamino)methyl)imidazo[1,5-a]pyridine-6-carboxylic acid ethyl ester (120mg, 0.21mmol), remove the dry ice after the addition, and continue to react for 3 Hour. After the reaction was completed, the reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product that was directly used in the next step.

Step 11: 5-(2-Fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-8-((4-(methylamino)-4-oxobutyloxyamino) Synthesis of Methyl)imidazo[1,5-a]pyridine-6-carboxamide

The crude product (120mg, 0.20mmol) obtained in the previous step was added to the reaction flask, methanol (2ml) was added, then 1M HCl (0.4ml, 0.4mmol) was added, and the reaction was carried out at room temperature for 30 minutes. After the reaction was completed, water was added, It was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the product was separated by column chromatography. (80mg, yield: 70%). [M+H] ⁺ ＝601.4

Step 12: 5-(2-Fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-8-((3-oxomorpholinyl)methyl)imidazo[1,5 -a] Synthesis of pyridine-6-carboxamide

Compound 5-(2-fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-8-((4-(methylamino)-4-oxobutyloxyamino)methyl Base) imidazo[1,5-a]pyridine-6-carboxamide (80mg, 0.13mmol), add to the reaction flask, add 1,2-dichloroethane (2ml), add to 60 ℃ for 10 hours After the reaction was detected by TLC, it was cooled to room temperature, concentrated, and separated by column chromatography to obtain the product. (40 mg, yield: 52%). ¹ HNMR (400MHz, CD ₃ OD) δ8.05(s,1H),7.45(dd,1H),7.41(dd,1H),7.28(s,1H),7.16(s,1H),6.51(m, 1H),4.86(s,2H),4.04(t,2H),3.92(t,2H),3.73(t,2H),2.53(t,2H),2.10(t,2H).m/z570(M +1).

Example 2: N-(2,3-hydroxypropoxy)-5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorpholine)methyl)imidazo[1 ,Synthesis of 5-a]pyridine-6-carboxamide

Step 1: Ethyl 5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorpholine)methyl)imidazo[1,5-a]pyridine-6-carboxylate synthesis

The compound 5-(2-fluoro-4-iodophenylamino)-8-((4-(methylamino)-4-oxobutoxyamino)methyl)imidazo[1,5-a] Add ethyl pyridine-6-carboxylate (500mg, 0.88mmol) into the reaction flask, add 1,2-dichloroethane (10ml), heat to 60°C and react for 10 hours, after the reaction is completed, cool to room temperature, Concentrate and separate the product by column chromatography. (400mg, yield: 84.6%). [M+H] ⁺ ＝539.3

Step 2: N-((2,2-Dimethyl-1,3-dioxolan-4-yl)methoxy)-5-(2-fluoro-4-iodophenylamino)-8- Synthesis of ((3-oxomorpholine)methyl)imidazo[1,5-a]pyridine-6-carboxamide:

Compound O-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)hydroxylamine (41mg, 0.28mmol) was added to the reaction flask, anhydrous THF (1ml) was added, Under nitrogen atmosphere, after cooling to -78°C, LiHMDS (1M, 0.56ml, 0.56mmol) was added dropwise to the reaction solution, and after reacting at -78°C for 30 minutes, the compound 5-(2-fluoro-4-iodo Phenylamino)-8-((3-oxomorpholine)methyl)imidazo[1,5-a]pyridine-6-carboxylic acid ethyl ester (100mg, 0.18mmol), remove the dry ice after addition, Continue to react for 3 hours. After the reaction was completed, the reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product that was directly used in the next step.

Step 3: N-(2,3-hydroxypropoxy)-5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorpholino)methyl)imidazo[1, 5-a] Synthesis of pyridine-6-carboxamide:

The compound N-((2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)-5-(2-fluoro-4-iodophenylamino)-8-( (3-oxomorpholine) methyl) imidazo[1,5-a]pyridine-6-carboxamide (118mg, 0.18mmol) was added to the reaction flask, then ethanol (1ml) and tetrahydrofuran (1ml) were added, and 1N hydrochloric acid (0.36ml, 0.36mmol) was added and reacted at room temperature for 2 hours. After the reaction was completed, it was concentrated and separated by column chromatography to obtain the compound (60mg, yield: 54%). ¹ HNMR (400MHz, CD ₃ OD) δ8.05(s,1H),7.43(dd,1H),7.39(dd,1H),7.28(s,1H),7.12(s,1H),6.49(m, 1H), 4.84(s, 2H), 4.04(t, 2H), 3.92(t, 2H), 3.73(t, 2H), 3.67(m, 1H), 2.51(t, 2H), 2.09(t, 2H ).m/z600.3(M+1)

Example 3: N-(cyclopropylmethyloxy)-5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorpholine)methyl)imidazo[1,5- a] the synthesis of pyridine-6-carboxamide:

Compound O-(cyclopropylmethyl)hydroxylamine (12mg, 0.14mmol) was added to the reaction flask, anhydrous THF (1ml) was added, and after cooling to -78°C under a nitrogen atmosphere, LiHMDS was added dropwise to the reaction solution (1M, 0.27ml, 0.27mmol), after reacting at -78°C for 30 minutes, add the compound 5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorpholine)methyl) Ethyl imidazo[1,5-a]pyridine-6-carboxylate (50mg, 0.09mmol), after the addition, the dry ice was removed, and the reaction was continued for 3 hours. After the reaction was completed, the reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain the product. (50 mg, yield: 55%). ¹ HNMR(400MHz, CD ₃ OD) ¹ HNMR(400MHz, CD ₃ OD)δ8.04(s,1H),7.45(dd,1H),7.41(dd,1H),7.24(s,1H),7.16( s,1H),6.51(m,1H),4.86(s,2H),3.87(d,2H),3.73(t,2H),2.53(t,2H),2.10(t,2H),0.65(m, 2H),0.41(m,1H),0.35(m,2H).m/z580.3(M+1)

Example 4: 5-(2-fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-8-((3-oxoisoxazol-2-yl)methyl)imidazole Synthesis of [1,5-a]pyridine-6-carboxamide:

Step 1: 5-(2-fluoro-4-iodophenylamino)-8-((3-(methylamino)-3-oxopropoxyamino)methyl)imidazo[1,5-a ] Synthesis of ethyl pyridine-6-carboxylate:

The compound 8-(bromomethyl)-5-(2-fluoro-4-iodophenylamino)imidazo[1,5-a]pyridine-6-carboxylic acid ethyl ester (150mg, 0.29mmol) was added to the reaction In the bottle, add acetonitrile (3ml), then add potassium carbonate (60mg, 0.43mmol) and 4-(aminooxy)-N-methylpropionamide (41mg, 0.43mmol), react at 60°C for 4 hours, cool Return to room temperature, add water to wash and extract with ethyl acetate, dry over anhydrous sodium sulfate, concentrate, and separate by column chromatography to obtain the product. (120 mg, yield: 73%). [M+H] ⁺ ＝556.4

Step 2: 5-(2-fluoro-4-iodophenylamino)-8-((3-oxoisoxazol-2-yl)methyl)imidazo[1,5-a]pyridine-6- Synthesis of ethyl carboxylate:

The compound 5-(2-fluoro-4-iodophenylamino)-8-((3-(methylamino)-3-oxopropoxyamino)methyl)imidazo[1,5-a] Ethyl pyridine-6-carboxylate (120mg, 0.21mmol) was added to the reaction flask, 1,2-dichloroethane (3ml) was added, and heated to 60°C for 10 hours. After the reaction was completed, the reaction solution was concentrated and columnar The product was isolated by chromatography. (90 mg, yield: 79%). [M+H] ⁺ ＝539.3

Step 3: 5-(2-Fluoro-4-iodophenylamino)-8-((3-oxoisoxazol-2-yl)methyl)-N-(2-(ethyleneoxy)ethoxy base) imidazo[1,5-a]pyridine-6-carboxamide

The compound O-(2-(vinyloxy)ethyl)hydroxylamine (26mg, 0.26mmol) was added to the reaction flask, anhydrous THF (1ml) was added, and after cooling to -78°C under a nitrogen atmosphere, the reaction LiHMDS (1M, 0.51ml, 0.51mmol) was added dropwise into the solution, and after reacting for 30 minutes at -78°C, the compound 5-(2-fluoro-4-iodophenylamino)-8-((4-(methyl Amino)-4-oxobutoxyamino)methyl)imidazo[1,5-a]pyridine-6-carboxylic acid ethyl ester (90mg, 0.17mmol), remove the dry ice after the addition, and continue to react for 3 Hour. After the reaction was completed, the reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product that was directly used in the next step. (100mg, yield: 100%)

Step 4: 5-(2-Fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-8-((3-oxoisoxazol-2-yl)methyl)imidazo [1,5-a]pyridine-6-carboxamide

Compound 5-(2-fluoro-4-iodophenylamino)-8-((3-oxoisoxazol-2-yl)methyl)-N-(2-(ethyleneoxy)ethoxy ) imidazo[1,5-a]pyridine-6-carboxamide (100mg, 0.17mmol) into the reaction flask, add methanol (2ml), then add 1M HCl (0.3ml, 0.3mmol), react at room temperature for 30 minutes , added water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain the product. (60 mg, yield: 62%). ¹ HNMR (400MHz, CD ₃ OD) δ8.05(s, 1H), 7.45(dd, 1H), 7.39(dd, 1H), 7.28(s, 1H), 7.13(s, 1H), 6.51(m, 1H ),4.76(s,2H),4.36(t,2H),3.93(t,2H),3.70(t,2H),2.84(t,2H),m/e556.30(M+1)

Example 5: 5-(2-fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-8-((3-oxomorpholine)methyl)-[1,2, 4] Synthesis of triazolo[4,3-a]pyridine-6-carboxamide

Step 1: Synthesis of ethyl 2-(2-fluoro-4-iodophenylamino)-6-hydrazino-5-methylnicotinate:

Compound 6-chloro-2-(2-fluoro-4-iodophenylamino)-5-methylnicotinic acid ethyl ester (5g, 11.5mmol) was added to the reaction flask, ethanol (30ml) was added, and then hydrazine hydrate was added 30% (1.35g, 12.65mmol), heated to 40°C and reacted for 4 hours. After the reaction, cooled to room temperature, concentrated, washed with water, extracted with dichloromethane, dried over anhydrous sodium sulfate, and concentrated to the target product. (4.2 g, yield: 85%). [M+H] ⁺ ＝431.2

Step 2: Synthesis of ethyl 2-(2-fluoro-4-iodophenylamino)-6-(2-formylhydrazino)-5-methylnicotinate:

Add the compound 2-(2-fluoro-4-iodophenylamino)-6-hydrazino-5-methylnicotinic acid ethyl ester (4.2g, into the reaction flask, add formic acid (30ml), cool to Add acetic anhydride (6ml) after 0°C, react at room temperature for 2 hours, concentrate the reaction solution, dissolve it with dichloromethane, wash with saturated sodium carbonate, and brine, dry over anhydrous sodium sulfate, and concentrate to obtain the target product. (4.4 g, yield: 98%)

Step 3: Synthesis of ethyl 5-(2-fluoro-4-iodophenylamino)-8-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylate :

Add the compound 2-(2-fluoro-4-iodophenylamino)-6-(2-formylhydrazino)-5-methylnicotinic acid ethyl ester (4.4g, 9.60mmol) into the reaction flask, add toluene ( 10ml), then add phosphorus oxychloride (2.94g, 19.20mmol), heat to 95 ° C for 1 hour, cool to room temperature, concentrate the residue obtained after dissolving with ethyl acetate, and wash with saturated sodium carbonate solution, saturated Wash with brine, dry the organic phase with anhydrous sodium sulfate, concentrate, and separate by column chromatography to obtain the target product. (2.1g, yield: 50%).[M+H] ⁺ =441.2

Step 4: 8-(bromomethyl)-5-(2-fluoro-4-iodophenylamino)-[1,2,4]triazolo[4,3-a]pyridine-carboxylic acid ethyl ester synthesis:

Compound 5-(2-fluoro-4-iodophenylamino)-8-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylic acid ethyl ester (2.10g , 4.77mmol) into the reaction flask, and then added carbon tetrachloride (20ml), NBS (0.94g, 5.26mmol) and benzoyl peroxide (BPO) (112mg, 0.47mmol). Heated to 60°C for 4 hours, cooled to room temperature, quenched with saturated sodium thiosulfate solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain the product. (1.8 g, yield: 72.6%). [M+H] ⁺ ＝520.1

Step 5: 5-(2-Fluoro-4-iodophenylamino)-8-((4-(methylamino)-4-oxobutoxyamino)methyl)-[1,2,4] Synthesis of ethyl triazolo[4,3-a]pyridine-6-carboxylate:

Compound 8-(bromomethyl)-5-(2-fluoro-4-iodophenylamino)-[1,2,4]triazolo[4,3-a]pyridine-carboxylic acid ethyl ester (500mg , 0.96mmol), added in the reaction flask, added acetonitrile (15ml), then added potassium carbonate (200mg, 1.44mmol) and 4-(aminooxy)-N-methylbutanamide (193mg, 1.44mmol), at 60 It was reacted at ℃ for 4 hours, cooled to room temperature, washed with water and extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain the product. (400 mg, yield: 73%). [M+H] ⁺ ＝570.4

Step 6: 5-(2-Fluoro-4-iodophenylamino)-8-((3-oxomorpholino)methyl)-[1,2,4]triazolo[4,3-a] Synthesis of ethyl pyridine-6-carboxylate:

The compound 5-(2-fluoro-4-iodophenylamino)-8-((4-(methylamino)-4-oxobutoxyamino)methyl)imidazo[1,5-a] Add ethyl pyridine-6-carboxylate (400mg, 0.70mmol) into the reaction flask, add 1,2-dichloroethane (10ml), heat to 60°C and react for 10 hours, after the reaction is completed, cool to room temperature, Concentrate and separate the product by column chromatography. (300 mg, yield: 79%). [M+H] ⁺ ＝540.3

Step 7: 5-(2-Fluoro-4-iodophenylamino)-8-((3-oxomorpholino)methyl)-N-(2-(vinyloxy)ethoxy)-[1 , Synthesis of 2,4]triazol[4,3-a]pyridine-6-carboxamide:

The compound O-(2-(vinyloxy)ethyl)hydroxylamine (29mg, 0.28mmol) was added to the reaction flask, anhydrous THF (1ml) was added, and after cooling to -78°C under a nitrogen atmosphere, the reaction LiHMDS (1M, 0.57ml, 0.57mmol) was added dropwise into the solution, and after reacting at -78°C for 30 minutes, the compound 5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorphol Phenyl)methyl)-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylic acid ethyl ester (100mg, 0.19mmol), remove the dry ice after the addition, and continue the reaction for 3 hours . After the reaction was completed, the reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product that was directly used in the next step.

Step 8: 5-(2-Fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-8-((3-oxomorpholino)methyl)-[1,2,4 Synthesis of ]triazolo[4,3-a]pyridine-6-carboxamide

Compound 5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorpholino)methyl)-N-(2-(vinyloxy)ethoxy)-[1, 2,4] Triazol[4,3-a]pyridine-6-carboxamide (100mg, 0.17mmol) was added to the reaction flask, methanol alkane (2ml) was added, then 1M HCl (0.34ml) was added, and the reaction was carried out at room temperature for 1 After hours, TLC detected that the reaction was complete, diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain the product. (50 mg, yield: 52%). ¹ HNMR (400MHz, CD ₃ OD) δ9.70(s,1H),7.45(dd,1H),7.41(dd,1H),7.16(s,1H),6.51(m,1H),4.86(s, 2H), 4.04(t, 2H), 3.92(t, 2H), 3.73(t, 2H), 2.53(t, 2H), 2.10(t, 2H). m/z 571(M+1).

Example 6: N-(2,3-hydroxypropoxy)-5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorpholine)methyl)imidazo[1 ,5-a]Synthesis of pyridine-6-carboxamide

Step 1: N-((2,2-dimethyl-1,3-dioxolan-4-ylmethoxy)-5-(2-fluoro-4-iodophenylamino)-8-( Synthesis of (3-oxomorpholine)methyl)-[1,2,4]triazolo[4,3-a]pyridine-6-carboxamide:

Compound O-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)hydroxylamine (41mg, 0.28mmol) was added to the reaction flask, anhydrous THF (1ml) was added, Under nitrogen atmosphere, after cooling to -78°C, LiHMDS (1M, 0.56ml, 0.56mmol) was added dropwise to the reaction solution, and after reacting at -78°C for 30 minutes, the compound 5-(2-fluoro-4-iodo Phenylamino)-8-((3-oxomorpholine)methyl)-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylic acid ethyl ester (100mg, 0.18mmol ), remove the dry ice after adding, and continue to react for 3 hours. After the reaction was completed, the reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product that was directly used in the next step.

Step 2: N-(2,3-hydroxypropoxy)-5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorpholino)methyl)imidazo[1, 5-a] Synthesis of pyridine-6-carboxamide:

The compound N-((2,2-dimethyl-1,3-dioxolan-4-ylmethoxy)-5-(2-fluoro-4-iodophenylamino)-8-(( 3-oxomorpholine) methyl)-[1,2,4]triazolo[4,3-a]pyridine-6-carboxamide (100mg, 0.16mmol) was added to the reaction flask, and then ethanol (1ml ) and tetrahydrofuran (1ml), then added 1N hydrochloric acid (0.32ml, 0.32mmol), and reacted at room temperature for 2 hours. After the reaction was completed, it was concentrated and separated by column chromatography to obtain compound (45mg, yield: 48%). ¹ HNMR(400MHz,CD ₃ OD)δ9.63(s,1H),7.43(dd,1H),7.39(dd,1H),7.12(s,1H),6.49(m,1H),4.84(s,2H ),4.04(t,2H),3.92(t,2H),3.73(t,2H),3.67(m,1H),2.51(t,2H),2.09(t,2H).m/z601.3( M+1)

Example 7: N-(cyclopropylmethyloxy)-5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorpholine)methyl)imidazo[1,5- a] the synthesis of pyridine-6-carboxamide:

Compound O-(cyclopropylmethyl)hydroxylamine (12mg, 0.14mmol) was added to the reaction flask, anhydrous THF (1ml) was added, and after cooling to -78°C under a nitrogen atmosphere, LiHMDS was added dropwise to the reaction solution (1M, 0.27ml, 0.27mmol), after reacting at -78°C for 30 minutes, add the compound 5-(2-fluoro-4-iodophenylamino)-8-((3-oxomorpholine)methyl) Ethyl imidazo[1,5-a]pyridine-6-carboxylate (50mg, 0.09mmol), after the addition, the dry ice was removed, and the reaction was continued for 3 hours. After the reaction was completed, the reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain the product. (28 mg, yield: 52%). ¹ HNMR(400MHz,CD ₃ OD) ¹ HNMR(400MHz,CD ₃ OD)δ9.64(s,1H),7.45(dd,1H),7.41(dd,1H),7.16(s,1H),6.51( m,1H),4.86(s,2H),3.87(d,2H),3.73(t,2H),2.53(t,2H),2.10(t,2H),0.65(m,2H),0.41(m, 1H),0.35(m,2H).m/z581.3(M+1)

biological example

Cell Viability Experiment

1. Cells: human colon cancer COLO205 and human melanoma A375 cells, both from the Basic Medical Cell Center of the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences.

2. Reagents: Gibco DMEM/F12 medium, Gibco 0.25% trypsin/EDTA cell digestion solution, MTT (5mg/ml), DMSO, PBS.

3. Instruments: 37°C, 5% CO ₂ incubator, TECANInfinite ^TM 200 series multifunctional microplate reader, ultra-clean bench, cell counting plate.

4. Experimental consumables: 96-well plate.

Experimental steps for the activity test of human colon cancer COLO205 cells:

1. Plank. Digest the cells in the logarithmic growth phase with digestion solution, terminate the fresh medium, count the cells, adjust the cell concentration to ⁵ *10 cells/ml with fresh medium, add 200 μL to each well, and set a zero well ( Only add medium) 3, the other edges are filled with sterile PBS.

2. Incubate at 37°C in 5% CO ₂ for 24 hours to allow the cells to cover about 60% of the bottom of the well.

3. Administration. Dissolve the drug in DMSO to make a 10mmol/L mother solution, and then dilute it with DMSO to make a 1mmol/L, 100μmol/L, 10μmol/L, 1mol/L, 0.1mol/L solution. When administering, take Dilute 1 μL of the above-mentioned concentration solution to 1 mL with medium, that is, the administration concentration is 10 μmol/L, 1 μmol/L, 100 nmol/L, 10 nmol/L, 1 nmol/L, 0.1 nmol/L, 0 nmol/L (control group, add 1 μL DMSO Dilute to 1ml with culture medium). When administering the drug, the liquid in the original wells was exhausted, and fresh medium containing different concentrations of drugs was added, 200 μl per well.

●Adjust the well for zero, only add the culture medium;

●The control group, containing the same volume of solvent as the experimental group, diluted with complete medium. 200 μl per well;

●Experimental group, the dissolved drug was diluted with culture medium to a concentration of 0.1, 1, 10, 100, 1000, 10000 nM, 200 μl per well.

4. Incubate at 37°C in 5% _CO2 .

After 5.72 hours, 20 μL of MTT solution (5 mg/ml) was added to each well, and culture was continued for 4 hours.

6. Centrifuge the 96-well plate with a plate centrifuge at 1000 rpm for 5 minutes.

7. Terminate the culture, carefully aspirate the culture medium in the well.

8. Add 150 μl dimethyl sulfoxide (DMSO) to each well, shake at a low speed for 10 minutes, and measure the absorbance value at a wavelength of 490 nm in a microplate reader after the crystals are fully dissolved.

The compounds of the present invention are numbered as shown in Table 2 below. All compounds 1-17 had _IC50 values less than 1000 nM.

Experimental steps for the activity test of human melanoma A375 cells:

1. Plank. Digest the cells in the logarithmic growth phase with digestion solution, terminate the fresh medium, count the cells, adjust the cell concentration to 2.5* ¹⁰ cells/ml with fresh medium, add 200 μL to each well, and set a zero well ( Only add medium) 3, the other edges are filled with sterile PBS.

2. Incubate at 37°C in 5% CO ₂ for 36 hours, let the cells cover about 60% of the bottom of the well.

3. Administration. Dissolve the drug in DMSO to make a 10mmol/L mother solution, and then dilute it with DMSO to make 1mmol/L, 100μmol/L, 10μmol/L, 1mol/L, 0.1mol/L solutions. When administering, take Dilute 1 μL of the above-mentioned concentration solution to 1 mL with medium, that is, the administration concentration is 10 μmol/L, 1 μmol/L, 100 nmol/L, 10 nmol/L, 1 nmol/L, 0.1 nmol/L, 0 nmol/L (control group, add 1 μL DMSO Dilute to 1ml with culture medium). When administering the drug, the liquid in the original wells was exhausted, and fresh medium containing different concentrations of drugs was added, 200 μl per well.

●Adjust the well for zero, only add the culture medium;

●The control group, containing the same volume of solvent as the experimental group, diluted with complete medium. 200 μl per well;

●Experimental group, the dissolved drug was diluted with culture medium to a concentration of 0.1, 1, 10, 100, 1000, 10000 nM, 200 μl per well.

4. Incubate at 37°C in 5% _CO2 .

After 5.72 hours, 20 μL of MTT solution (5 mg/ml) was added to each well, and culture was continued for 4 hours.

6. Terminate the culture, and carefully aspirate the culture medium in the well.

7. Add 150 μl dimethyl sulfoxide (DMSO) to each well, shake at a low speed for 10 minutes, and measure the absorbance value at a wavelength of 490 nm in a microplate reader after the crystals are fully dissolved.

The compounds of the present invention are numbered as shown in Table 1 below. All compounds 1-7 had IC ₅₀ values less than 1000 nM.

Table 1. Test Compounds

Claims (14)

1. the compound of formula (I) and (II) and pharmacy acceptable salt, prodrug and solvate

Wherein

A is selected from C and N;

R ¹be selected from hydrogen, halogen, cyano group, nitro, azido-, hydroxyl, C ₁-C ₁₀alkoxyl group, halo C ₁-C ₁₀alkoxyl group, sulfydryl, C ₁-C ₁₀alkylthio, halo C ₁-C ₁₀alkylthio, carboxyl ,-OC (O) H, amino, C ₁-C ₁₀alkyl, C ₂-C ₁₀thiazolinyl, C ₂-C ₁₀alkynyl, C ₃-C ₁₀cycloalkyl;

R ²be selected from hydrogen, halogen, nitro, azido-, hydroxyl, C ₁-C ₁₀alkyl, C ₁-C ₁₀alkoxyl group, halo C ₁-C ₁₀alkoxyl group, C ₁-C ₁₀alkylthio, halo C ₁-C ₁₀alkylthio, carboxyl ,-OC (O) H, amino, C ₂-C ₁₀thiazolinyl, C ₂-C ₁₀alkynyl, C ₃-C ₁₀cycloalkyl;

R ₃be selected from hydrogen, C ₁-C ₁₂alkyl, C ₂-C ₁₂thiazolinyl, C ₂-C ₁₂alkynyl, C ₃-C ₁₂cycloalkyl, C ₃-C ₁₂cycloalkyl C ₁-C ₁₂alkyl, C ₁-C ₁₂alkyl C ₃-C ₁₂cycloalkyl, described alkyl, thiazolinyl, alkynyl and cycloalkyl can be selected from following group by one to three independently of one another and replace arbitrarily :-ORa ,-NRaRb ,-NRaC (O) Rb, Heterocyclylalkyl and heterocyclic aromatic base, and Heterocyclylalkyl described herein and heterocyclic aromatic base can be selected from following substituting group by one to three and replace arbitrarily: C ₁-C ₅alkyl ,-ORa ,-NRaRb;

X representative formula (i):

Wherein

Y is selected from O ,-ONR ₈,-NR ₈o ,-NR ₈c (O) ,-NR ₈sO ₂;

Z is selected from hydrogen, C ₁-C ₅methene chain, described methylene radical can be replaced arbitrarily by one to three W';

R ₈be selected from hydrogen, C ₁-C ₁₂alkyl ,-COR ₉, described alkyl can be replaced arbitrarily by halogen ,-ORa ,-NRaRb;

R ₉be selected from hydrogen, C ₁-C ₁₂alkyl, described alkyl can be replaced arbitrarily by halogen ,-ORa ,-NRaRb;

Or X representative formula (ii):

Wherein

Y ₁and Y ₂can be the same or different, they represent singly-bound ,-CO-,-COO-,-O-,-OCO-,-NRa-,-SO independently of one another ₂-;

Z' represents C ₁-C ₁₀chain alkylene, described alkyl can be replaced arbitrarily by one to three W';

In above-mentioned formula (i) and formula (ii),

W and W' can be the same or different, and they are selected from halogen, C independently of one another ₁-C ₅alkyl ,-O-,-ORa ,-COORa ,-COOCORa ,-CO-halogen ,-OCORa ,-CONRaRb ,-SRa ,-SORa ,-SO ₂ra ,-NRaRb ,-NRaCORb ,-NRaSO ₂rb ,-SO ₂nRaRb, Heterocyclylalkyl or heterocyclic aromatic base, described Heterocyclylalkyl and heterocyclic aromatic base can be selected from following substituting group by one to three and replace arbitrarily: C ₁-C ₅alkyl ,-ORa ,-NRaRb; Described alkyl can by hydroxyl, C ₁-C ₅alkoxyl group or amino replacement arbitrarily;

Above-mentioned group deoxygenation and halogen all can be interconnected to form cycloalkyl or Heterocyclylalkyl outward, and described cycloalkyl or Heterocyclylalkyl can be selected from following substituting group and replace arbitrarily :-ORa ,-NRaRb, the C replaced arbitrarily by-ORa ₁-C ₅alkyl;

Ra and Rb can be the same or different, and they are selected from hydrogen, C independently of one another ₁-C ₅alkyl, described alkyl can be selected from following substituting group by one to three and replace arbitrarily: hydroxyl, C ₁-C ₅alkoxyl group, amino.

2. the compound of claim 1 and pharmacy acceptable salt, prodrug and solvate, wherein X representative formula (i):

3. the compound of claim 2 and pharmacy acceptable salt, prodrug and solvate, wherein

Y is selected from-O-,-NR ₈o-,-NR ₈c (O)-,-NR ₈sO ₂-;

Z is selected from hydrogen ,-CH ₂-,-(CH ₂) ₂-,-(CH ₂) ₃-,-CH ₂(CH ₃) CH ₂-,-CH ₂c (CH ₃) ₂-,-CH ₂c (CH ₃) ₂cH ₂-,-CH ₂c (CH ₂cH ₂) CH ₂-;

R ⁸be selected from hydrogen, methyl ,-COH ,-COCH ₃,-COCH ₂cH ₃,-COCH (CH ₃) ₂,-COCH ₂oH;

W and W' can be the same or different, and they are selected from hydrogen, hydroxyl, methyl ,-O (CH independently of one another ₂) ₂oH ,-C (O) OCH ₃,-CONHCH ₃,-CONHCH ₂cH ₃,-CONHCH (CH ₃) ₂,-CON (CH ₃) ₂,-NHCH ₃,-NH (CH ₂) ₂oH ,-NHCOCH ₃,-NHCOCH ₂cH ₃,-NHCOCH (CH ₃) ₂,-SCH ₃,-SOCH ₃, 1H-imidazoles-2-base, morpholine-4-base, 4-hydroxy piperidine base.

4. the compound of claim 1 and pharmacy acceptable salt, prodrug and solvate, wherein X representative formula (ii):

5. the compound of claim 4 and pharmacy acceptable salt, prodrug and solvate, wherein X is selected from

Z' is selected from C ₁-C ₃chain alkylene, described alkyl can be replaced arbitrarily by one to three W';

W' is selected from C ₁-C ₃alkyl ,-ORa ,-COORa ,-COOCORa ,-CO-halogen ,-OCORa ,-CONRaRb ,-SRa ,-SORa ,-SO ₂ra ,-NRaRb ,-NRaCORb ,-NRaSO ₂rb ,-SO ₂nRaRb;

Ra and Rb can be the same or different, and they are selected from hydrogen, C independently of one another ₁-C ₃alkyl, described alkyl can be selected from following substituting group by one to three and replace arbitrarily: hydroxyl, C ₁-C ₃alkoxyl group, amino.

6. the compound of claim 5 and pharmacy acceptable salt, prodrug and solvate, wherein X is selected from

Z' is selected from-CH ₂-,-(CH ₂) ₂-,-(CH ₂) ₃-,-(CH ₂) ₄-, wherein hydrogen atom is all replaced arbitrarily by one to three W' arbitrarily;

W' is selected from hydroxyl, C ₁-C ₂alkyl.

7. the compound of claim 6 and pharmacy acceptable salt, prodrug and solvate, wherein X is selected from

8. arbitrary compound of claim 1-7 and pharmacy acceptable salt, prodrug and solvate, wherein

A is selected from C and N;

R ₁be selected from fluorine, chlorine, bromine, iodine, C ₁-C ₄alkoxyl group, C ₁-C ₄alkylthio, halo-C ₁-C ₄alkoxyl group, halo-C ₁-C ₄alkylthio, C ₁-C ₄alkyl, halo C ₁-C ₄alkyl C ₂-C ₄thiazolinyl, C ₂-C ₄alkynyl;

R ₂be selected from hydrogen, halogen or C ₁-C ₄alkyl;

R ₃for C that is unsubstituted or that replaced by 1 to 6 hydroxyl ₁-C ₄alkyl, C ₃-C ₄cycloalkyl, C ₃-C ₄cycloalkyl C ₁-C ₄alkyl or C ₁-C ₄alkyl C ₃-C ₄cycloalkyl.

9. the compound of claim 8 and pharmacy acceptable salt, prodrug and solvate, wherein

R ₁be selected from bromine, iodine, C ₁-C ₂alkylthio, halo-C ₁-C ₂alkylthio, C ₁-C ₂alkoxyl group, halo-C ₁-C ₂alkoxyl group, C ₁-C ₂alkyl, halo C ₁-C ₂alkyl, C ₂-C ₃thiazolinyl, C ₂-C ₃alkynyl;

R ₂be selected from hydrogen, fluorine, chlorine, bromine or C ₁-C ₂alkyl;

R ₃for C that is unsubstituted or that replaced by 1 to 3 hydroxyl ₁-C ₃alkyl, C ₃-C ₄cycloalkyl, C ₃-C ₄cycloalkyl C ₁-C ₃alkyl or C ₁-C ₃alkyl C ₃-C ₄cycloalkyl.

10. the compound of claim 9 and pharmacy acceptable salt, prodrug and solvate, wherein

R ₁be selected from bromine, iodine, methylthio group, trifluoromethylthio, methoxyl group, trifluoromethoxy, methyl, trifluoromethyl, vinyl, ethynyl;

R ₂be selected from fluorine, chlorine or methyl;

R ₃be selected from 2-hydroxyethyl, 3-hydroxyethyl, 2,3-dihydroxypropyls, 1-methylol-2-hydroxyethyl, 2-methyl-3-hydroxypropyl, cyclopropyl, Cvclopropvlmethvl or 1-(2,3-hydroxypropyl) cyclopropyl.

Formula (I) compound of 11. claims 1, and pharmacy acceptable salt, prodrug and solvate, wherein said compound is selected from:

The preparation method of formula (I) compound of 12. claims 1, comprising:

(A) route 1

(B) route 2

(C) formula (I-1-a) compound is prepared according to following route 3:

Wherein, the synthesized reference route 2 of compound 11

(D) formula (I-1-b) compound is prepared according to following route 4:

Wherein, the synthesized reference route 2 of compound 11

(E) formula (I-1-c) compound is prepared according to following route 5:

(F) formula (I-2-A) compound is prepared according to following route 6:

(G) formula (I-2-A) compound is prepared according to following route 7:

13. 1 kinds of medicinal compositionss comprising formula (I) and (II) compound, its pharmacy acceptable salt, prodrug and/or solvate.

14. formulas (I) and (II) compound and pharmacy acceptable salt, prodrug and solvate for the manufacture of the mammiferous tumour for the treatment of, chronic inflammatory diseases, inflammatory bowel disease, tetter, diabetes, eye disease, with the disease that mammiferous blood vessel occurs or revascularization is relevant, and the purposes of medicine of the chronic pain disease of being correlated with and other disease of being modulated by Mek cascade.