WO2017148440A1 - Pteridinone derivative serving as flt3 inhibitor, and uses - Google Patents

Abstract

The present invention relates to a pteridinone derivative serving as an FLT3 inhibitor, and uses thereof. Specifically, the present invention relates to a compound represented by formula (I), a pharmaceutical composition containing the compound represented by formula (I), and uses of the compound in the preparation of drugs for treating diseases mediated by FLT3 or for inhibiting the FLT3: (I).

Description

Phetidone derivatives and their applications as FLT3 inhibitors Technical field

The present invention relates to the field of medicinal chemistry; in particular, the present invention relates to novel pteridine derivatives, methods for their synthesis and their use as FLT3 inhibitors in the preparation of medicaments for tumors and immune diseases.

Background technique

Protein tyrosine kinase (protein tyrosine kinase) is a class of proteins that catalyze the transfer of γ-phosphate on ATP to specific amino acid residues of proteins. It plays a very important role in the signal transduction pathway in cells and regulates cell growth. A series of physiological processes such as differentiation and death. It has been shown that more than 50% of protooncogenes and their products have protein tyrosine kinase activity, and their abnormal expression will lead to cell cycle life disorder, leading to tumorigenesis. In addition, the abnormal expression of tyrosine kinase is also closely related to tumor metastasis, chemotherapy resistance and the like.

FMS-like tyrosine kinase 3 (FLT3) belongs to the family of type III receptor tyrosine kinases. FLT3 plays an important role in the proliferation, differentiation and apoptosis of hematopoietic cells (Oncogene, 1993, 8,815-822). Upon binding to FLT3 ligand, FLT3 activates multiple downstream signaling pathways, including STAT5, Ras/MAPK, and PI3K/AKT pathways. There is a FLT3 mutation in approximately one-third of patients with acute myeloid leukemia (AML) (Blood, 2002, 100, 1532-1542), including the proximal membrane domain 14 and/or exon 15 An internal tandem repeat (FLT3-ITD) mutation, an amino acid deletion or insertion (FLT3-TKD) mutation in the activation loop of the tyrosine kinase domain. In addition, there is a high expression of FLT3 in acute leukemia cases (Blood, 2004, 103, 1901), FLT3 overexpression, FLT3-ITD mutations and FLT3-TKD mutations can lead to poor prognosis in patients with AML. Therefore, FLT3 is an important target for AML treatment. To date, no FLT3 inhibitors have been approved for clinical use, and the clinical effects of many FLT3 inhibitors in clinical trials are still not ideal.

Therefore, improving the clinical efficiency of small molecule kinase inhibitors is becoming a hot spot in the development of anti-tumor targeted drugs. The most promising strategy is to develop multiple targets that simultaneously target multiple kinases associated with disease (tumor). Inhibitor.

Summary of the invention

It is an object of the present invention to provide a compound capable of inhibiting FLT3, which compound or a pharmaceutical composition containing the same can be used for the preparation of a medicament for treating a FLT3-mediated disease or inhibiting FLT3.

In a first aspect, the present invention relates to a compound having the structure of Formula I or a pharmaceutically acceptable salt thereof:

In the formula,

X is selected from: N, CH;

Y is selected from the group consisting of: O, S, and Se;

A is selected from various substituted benzene rings, nitrogen-containing five-membered rings, nitrogen-containing six-membered rings, for example

R ^{1 is} independently selected from the group consisting of hydrogen, halogen, C ₁ -C ₈ alkyl, C ₃ -C ₆ cycloalkyl, fluorine-containing C ₁ -C ₈ alkyl, optionally substituted phenyl;

R ^{2 is} selected from the group consisting of hydrogen, optionally substituted C ₁ -C ₆ alkyl, halogen, optionally substituted C ₁ -C ₆ alkoxy, amino, -NR _a R _b , carboxyl, optionally substituted alkoxy Formyl, optionally substituted N-alkylpiperazin-1-yl, optionally substituted morpholin-4-yl, optionally substituted piperidin-1-yl, optionally substituted pyrrol-1-yl, Optionally substituted pyrrolidin-1-yl, optionally substituted aryl or optionally substituted arylmethyl, optionally substituted piperidinyloxy, optionally substituted N,N',N'-three Alkyl ethylenediamine;

R _a and R _b are each independently selected from the group consisting of: hydrogen, C ₁ -C ₆ alkyl and alkenyl;

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

R ^{3 is} each independently selected from the group consisting of hydrogen, halogen, hydroxy, amino, -NR _c R _d , C ₁ -C ₂ acylamino, optionally substituted C ₁ -C ₆ alkyl, C ₁ -C ₃ sulfonylamino, CN, carbamoyl, carboxyl, C1-C6 alkoxycarbonyl, optionally substituted phenyl, optionally substituted piperazinyl, optionally substituted morpholin-4-yl, optionally substituted pyrrol-1- Base, optionally substituted pyrrolidin-1-yl;

R _c and R _d are each independently selected from the group consisting of: hydrogen, C ₁ -C ₆ alkyl and alkenyl;

m is 0, 1, 2, 3 or 4;

Wherein, when both n and m are 1, and R ² is a C ₁ -C ₆ alkoxy group, R ^{3 is} not an amino group.

In a specific embodiment, the compound has the structure shown in Formula II:

In the formula,

A is selected from various substituted benzene rings;

R ¹ is hydrogen;

R ^{2 is} selected from the group consisting of hydrogen, optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₁ -C ₆ alkoxy, halogen, amino, -NR _a R _b , optionally substituted piperazinyl, optionally Substituted N-alkylpiperazin-1-yl, optionally substituted piperidinyloxy;

R _a and R _b are each independently selected from the group consisting of: hydrogen, C1-C6 alkyl and alkenyl;

n is 1, 2, or 3;

R ^{3 is} each independently selected from the group consisting of: hydrogen, halogen, C1-C2 acylamino, hydroxy, amino, -NR _c R _d , methanesulfonylamino, optionally substituted C ₁ -C ₆ alkyl, optionally substituted piperazine base;

R _c and R _d are each independently selected from the group consisting of: hydrogen, C ₁ -C ₃ alkyl and alkenyl;

m is 1, 2, or 3.

In a specific embodiment, the compound has the structure shown in Formula III:

In the formula,

R ^{4 is} selected from the group consisting of hydrogen, halogen, optionally substituted C ₁ -C ₆ alkoxy, optionally substituted C ₁ -C ₆ alkyl;

R ^{5 is} selected from the group consisting of: an optionally substituted N-alkylpiperazin-1-yl, optionally substituted piperidinyloxy;

R ^{6 is} selected from the group consisting of: an amino group, an optionally substituted piperazinyl group, -NR _e R _f ;

R _e and R _f are each independently selected from the group consisting of hydrogen, C ₁ -C ₃ alkyl and alkenyl.

In a specific embodiment, R ^{4 is} selected from an optionally substituted C ₁ -C ₆ alkyl group, preferably a C ₁ -C ₃ alkyl group; and R ^{5 is} selected from an optionally substituted N-alkylpiperazin-1-yl group. R ^{6 is} selected from an amino group, or -NR _e R _f ; and R _e and R _f are each independently selected from hydrogen, and a C ₁ -C ₃ alkyl group.

In a second aspect, the invention provides a specific compound selected from the group consisting of: or a pharmaceutically acceptable salt thereof:

In a third aspect, the present invention provides a pharmaceutical composition comprising the compound of the first or second aspect of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or Shape agent.

In a fourth aspect, the invention provides the use of a compound of the first or second aspect of the invention for the manufacture of a medicament for the treatment of a FLT3-mediated disease, or for the inhibition of FLT3.

In a specific embodiment, the FLT3 mediated disease is a blood disorder, such as a myeloproliferative disorder, a cancer, an immune disease, and a skin disorder such as psoriasis and atopic dermatitis.

In a further embodiment, the cancer is selected from the group consisting of acute myeloid leukemia (AML), mixed line leukemia (MLL), T-cell acute leukemia (T-ALL), B-cell acute leukemia (B-ALL), chronic Bone marrow monocytic leukemia (CMML), chronic lymphocytic leukemia, chronic myeloid leukemia, chronic neutrophilic leukemia.

In a further embodiment, the immune disease is selected from the group consisting of arthritis, lupus, inflammatory bowel disease, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis , diabetes, myasthenia gravis, Hashimoto's thyroiditis, Odd thyroiditis, Graves' disease, rheumatoid arthritis syndrome, multiple sclerosis, infectious neuronitis, acutely transmitted encephalomyelitis, Dyson disease, aplastic anemia, autoimmune hepatitis, optic neuritis, silvery disease, graft versus host disease, transplantation, blood transfusion allergy, allergy, type I hypersensitivity, allergic conjunctivitis, allergic rhinitis Atopic dermatitis.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

DRAWINGS

Figure 1A shows that Western blot analysis determined that compound F-8 autophosphorylates FLT3 and inhibits phosphorylation of downstream signaling pathways; Figure 1B shows compound 20 induction at various concentrations over 48 hours of cell culture. MV4-11 cell apoptosis; Figure 1C shows inhibition of G1 phase cell growth with a specific concentration of compound 20 after 48 hours of culture;

Figure 2 shows tumor volume changes in the MV4-11 xenograft model;

Figure 3 shows tumor weight changes in the MV4-11 xenograft model.

detailed description

The inventors After extensive and intensive study, has unexpectedly been found a new group of the structure of pteridine derivatives, these derivatives to inhibit the activity value ₅₀ nM IC FLT3 level reached; the IC ₅₀ values of inhibitory activity related to the cell reaches μM level. The present invention has been completed on this basis.

The present inventors synthesized a series of pteridinone derivatives which have not been reported in the literature with FLT3 inhibitory activity, and structurally characterized these compounds. A series of compounds were tested for activity at the molecular and cellular levels to obtain a batch of compounds having FLT3 inhibition activity. Among them, a plurality of compounds, such as F-8 and F-16, have an inhibitory activity against FLT3 with an IC ₅₀ value of less than 10 nM, which is far superior to the positive control MLN 518.

Definition of Terms

Some of the terms involved in this article are explained as follows:

As used herein, "alkyl" refers to a saturated branched or straight-chain alkyl group having a carbon chain length of from 1 to 10 carbon atoms, and preferred alkyl groups include from 2 to 8 carbon atoms, from 1 to 6, and from 1 to 4 carbon atoms. An alkyl group having carbon atoms, 3-8 carbon atoms, and 1-3 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, heptyl, and the like. The alkyl group of the present invention may be substituted with one or more substituents, for example, by halogen or haloalkyl. For example, an alkyl group can be 1-4 The fluorine atom-substituted alkyl group or the alkyl group may be an alkyl group substituted by a fluoroalkyl group.

As used herein, "alkoxy" refers to an oxy group substituted with an alkyl group. Preferred alkoxy groups are alkoxy groups having from 1 to 6 carbon atoms, more preferably alkoxy groups having from 1 to 4 carbon atoms. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, and the like.

As used herein, "alkenyl" generally denotes a monovalent hydrocarbon radical having at least one double bond, usually containing from 2 to 8 carbon atoms, preferably from 2 to 6 carbon atoms, and may be straight or branched. Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, hexenyl, and the like.

As used herein, "halogen atom" or "halogen" means fluoro, chloro, bromo and iodo.

"Aryl" means a monocyclic, bicyclic or tricyclic aromatic group containing from 6 to 14 carbon atoms, including phenyl, naphthyl, phenanthryl, anthracenyl, fluorenyl, fluorenyl, tetrahydronaphthyl, Indane or the like. The aryl group may be optionally substituted with from 1 to 5 (eg, 1, 2, 3, 4 or 5) substituents selected from the group consisting of halogen, C1-4 aldehyde group, C1-6 alkyl group, cyano group, Nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (eg trifluoromethyl), halogen substituted alkoxy (eg trifluoromethoxy), carboxyl, C1-4 alkoxy, ethoxy a formyl group, a N(CH ₃ ) group, a C1-4 acyl group or the like, a heterocyclic group or a heteroaryl group.

As used herein, "aralkyl" refers to an alkyl group substituted with an aryl group, such as a C1-C6 alkyl group substituted with a phenyl group. Examples of aralkyl groups include, but are not limited to, arylmethyl, arylethyl, and the like, such as benzyl, phenethyl, and the like.

For example, an aryl group may be substituted with from 1 to 3 groups selected from the group consisting of halogen, -OH, C1-4 alkoxy, C1-4 alkyl, -NO ₂ , -NH ₂ , -N(CH ₃ ) ₂ , carboxyl and ethoxylated groups.

"5- or 6-membered heterocyclic ring" as used herein includes, but is not limited to, a heterocyclic group containing 1-3 heteroatoms selected from O, S and N, including but not limited to furyl, thienyl, pyrrolyl, Pyrrolidinyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, pyranyl, pyridyl, pyrimidinyl, pyrazinyl, piperidinyl, morpholinyl and the like.

As used herein, "heteroaryl" refers to 5-14 ring atoms and 6, 10 or 14 electrons are shared on the ring system. Further, the ring atom contained is a carbon atom and optionally 1-3 hetero atoms from oxygen, nitrogen, and sulfur. Heteroaryl groups useful in the present invention include piperazinyl, morpholinyl, piperidinyl, pyrrolidinyl, thienyl, furyl, pyranyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, including but It is not limited to 2-pyridyl, 3-pyridyl and 4-pyridyl, pyrazinyl, pyrimidinyl and the like.

The heteroaryl or 5- or 6-membered heterocyclic ring may be optionally substituted with from 1 to 5 (eg, 1, 2, 3, 4 or 5) substituents selected from halogen, C1-4 aldehyde, C1-6 straight or branched alkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (eg trifluoromethyl), halogen substituted alkoxy (eg trifluoromethoxy) Base), carboxyl group, C1-4 alkoxy group, ethoxylated group, N(CH ₃ ) and C1-4 acyl group.

As used herein, "acyloxy" refers to a radical of the formula "-O-C(O)-R", wherein R may be selected from alkyl, alkenyl and alkynyl. The R can be optionally substituted.

As used herein, "amido" refers to a group of the formula "-R'-NH-C(O)-R", wherein R' may be selected from a bond or an alkyl group, and R may be selected from an alkyl group, an alkenyl group. , an alkynyl group, NR _a R _b is a substituted alkyl group, NR _a R _b is a substituted alkenyl and NR _a R _b substituted alkynyl, halogen substituted alkyl, alkenyl substituted with cyano group,

Wherein R _a and R _b may be selected from the group consisting of alkyl and alkenyl.

As used herein, "optionally substituted" means that the substituent to which it is modified may be optionally substituted with from 1 to 5 (eg, 1, 2, 3, 4 or 5) substituents selected from halogen: C1 a 4-aldehyde group, a C1-6 straight or branched alkyl group, a cyano group, a nitro group, an amino group, a hydroxyl group, a hydroxymethyl group, a halogen-substituted alkyl group (for example, a trifluoromethyl group), a halogen-substituted alkoxy group ( For example, trifluoromethoxy), carboxyl, C1-4 alkoxy, ethoxycarbonyl, N(CH ₃ ), and C1-4 acyl.

Compound of the invention

The compound of the present invention is a compound having the structure of the formula I or a pharmaceutically acceptable salt thereof:

In the formula,

X is selected from N, CH;

Y is selected from the group consisting of O, S and Se;

A is selected from various substituted benzene rings, nitrogen-containing five-membered rings, nitrogen-containing six-membered rings, for example

R ^{1 is} independently selected from the group consisting of hydrogen, halogen, C ₁ -C ₈ alkyl, C ₃ -C ₆ cycloalkyl, fluoro C ₁ -C ₈ alkyl, optionally substituted phenyl;

R ^{2 is} selected from hydrogen, optionally substituted C ₁ -C ₆ alkyl, halogen, optionally substituted C ₁ -C ₆ alkoxy, amino, —NR _a R _b , carboxy, optionally substituted alkoxylate Acyl, optionally substituted N-alkylpiperazin-1-yl, optionally substituted morpholin-4-yl, optionally substituted piperidin-1-yl (preferably piperazine substituted piperidin-1-yl) , optionally substituted pyrrol-1-yl, optionally substituted pyrrolidin-1-yl, optionally substituted aryl or optionally substituted arylmethyl, optionally substituted piperidinyloxy, any Substituted substituted N,N',N'-trialkylethylenediamine;

R _a and R _b are each independently selected from the group consisting of hydrogen, C ₁ -C ₆ alkyl and alkenyl;

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

R ^{3 is} each independently selected from the group consisting of hydrogen, halogen, hydroxy, amino, -NR _c R _d , C1-C2 acylamino, optionally substituted C ₁ -C ₆ alkyl, C ₁ -C ₃ sulfonylamino, CN, amino Formyl, carboxyl, C1-C6 alkoxycarbonyl, optionally substituted phenyl, optionally substituted piperazinyl, optionally substituted morpholin-4-yl, optionally substituted pyrrol-1-yl, any Substituted pyrrolidin-1-yl;

R _c and R _d are each independently selected from the group consisting of: hydrogen, C ₁ -C ₆ alkyl and alkenyl;

m is 0, 1, 2, 3 or 4;

Wherein, when both n and m are 1, and R ² is a C ₁ -C ₆ alkoxy group, R ^{3 is} not an amino group.

In a specific embodiment, the compound has the structure shown in Formula II:

In the formula,

A is selected from various substituted benzene rings;

R ¹ is hydrogen;

R ^{2 is} selected from hydrogen, optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₁ -C ₆ alkoxy, halogen, amino, -NR _a R _b , optionally substituted piperazinyl, optionally substituted N-alkylpiperazin-1-yl, optionally substituted piperidinyloxy;

R _a and R _b are each independently selected from the group consisting of hydrogen, C1-C6 alkyl and alkenyl;

n is 1, 2, or 3;

R ^{3 is} each independently selected from the group consisting of hydrogen, halogen, C1-C2 acylamino, hydroxy, amino, -NR _c R _d , C1-C3 sulfonylamino (preferably methanesulfonylamino), optionally substituted C ₁ -C ₆ alkane , optionally substituted piperazinyl, optionally substituted N-alkylpiperazin-1-yl;

R _c and R _d are each independently selected from the group consisting of hydrogen, C ₁ -C ₃ alkyl and alkenyl;

m is 1, 2, or 3.

In a preferred embodiment, the compound has the structure shown in Formula III:

In the formula,

R ^{4 is} selected from the group consisting of hydrogen, halogen, optionally substituted C ₁ -C ₆ alkoxy, optionally substituted C ₁ -C ₆ alkyl;

R ^{5 is} selected from the group consisting of an optionally substituted N-alkylpiperazin-1-yl, optionally substituted piperidinyloxy;

R ^{6 is} selected from the group consisting of amino, optionally substituted piperazinyl, -NR _e R _f ;

R _e and R _f are each independently selected from the group consisting of hydrogen, C ₁ -C ₃ alkyl and alkenyl.

In a further preferred embodiment:

R ^{4 is} selected from an optionally substituted C ₁ -C ₆ alkyl group, preferably a C ₁ -C ₃ alkyl group;

R ^{5 is} selected from the group consisting of an optionally substituted N-alkylpiperazin-1-yl;

R ^{6 is} selected from amino, or -NR _e R _f ;

R _e and R _f are each independently selected from the group consisting of hydrogen and C ₁ -C ₃ alkyl.

The present inventors synthesized a series of pteridine ketone compounds which have not been reported in the literature; specifically, the present inventors synthesized a compound selected from the group consisting of or a pharmaceutically acceptable salt thereof:

In a preferred embodiment, the inventors synthesize a compound selected from the group consisting of: or a pharmaceutically acceptable salt thereof:

In a further preferred embodiment, the inventors synthesize a compound selected from the group consisting of or a pharmaceutically acceptable salt:

Based on the compounds of the present invention, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient .

Examples of pharmaceutically acceptable salts of the compounds of the invention include, but are not limited to, inorganic and organic acid salts such as the hydrochloride, hydrobromide, sulfate, citrate, lactate, tartrate, maleate salts. , fumarate, mandelate and oxalate; and inorganic and formed with bases such as sodium hydroxy, tris(hydroxymethyl)aminomethane (TRIS, tromethamine) and N-methyl glucosamine Organic base salt.

While the needs of each individual vary, one skilled in the art can determine the optimal dosage of each active ingredient in the pharmaceutical compositions of the present invention. In general, the compound of the present invention or a pharmaceutically acceptable salt thereof is orally administered to a mammal daily in an amount of from about 0.0025 to 50 mg/kg body weight. Preferably, however, it is about 0.01 to 10 mg per kilogram of oral administration. For example, a unit oral dose can include from about 0.01 to 50 mg, preferably from about 0.1 to 10 mg, of a compound of the invention. The unit dose may be administered one or more times per day in one or more tablets, each tablet containing from about 0.1 to 50 mg, conveniently from about 0.25 to 10 mg of the compound of the invention or a solvate thereof.

The pharmaceutical composition of the present invention can be formulated into a form suitable for various administration routes, including but not limited to being formulated for parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, intrathecal, intracranial A form of administration, intranasal or topical, for the treatment of tumors and other diseases. The amount administered is an amount effective to ameliorate or eliminate one or more conditions. For the treatment of a particular disease, an effective amount is an amount sufficient to ameliorate or in some way alleviate the symptoms associated with the disease. Such doses can be administered as a single dose or can be administered according to an effective therapeutic regimen. The amount administered may cure the disease, but administration is usually to improve the symptoms of the disease. Repeated administration is generally required to achieve the desired improvement in symptoms. The dosage of the drug will be determined by the age of the patient, the health and weight, the type of concurrent treatment, the frequency of treatment, and the desired therapeutic benefit.

The pharmaceutical preparation of the present invention can be administered to any mammal as long as they can obtain the therapeutic effect of the compound of the present invention. The most important of these mammals is humans.

The compounds of the invention or pharmaceutical compositions thereof are useful in the treatment of a variety of diseases mediated by FMS-like tyrosine kinase 3 (FLT3). Herein, the FLT3-mediated diseases are blood diseases such as myeloproliferative diseases, cancers, immune diseases, and skin diseases such as psoriasis and atopic dermatitis.

In a specific embodiment, the cancer is selected from the group consisting of acute myeloid leukemia (AML), mixed line leukemia (MLL), T-cell acute leukemia (T-ALL), B-cell acute leukemia (B-ALL), chronic bone marrow Single cell leukemia (CMML), chronic lymphocytic leukemia, chronic myeloid leukemia, chronic neutrophilic leukemia.

In other embodiments, the immune disease is selected from the group consisting of arthritis, lupus, inflammatory bowel disease, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes , myasthenia gravis, Hashimoto's thyroiditis, Odd thyroiditis, Graves' disease, rheumatoid arthritis syndrome, multiple sclerosis, infectious neuronitis, acutely transmitted encephalomyelitis, Addison Disease, aplastic anemia, autoimmune hepatitis, optic neuritis, silver disease, graft versus host disease, transplantation, blood transfusion allergy, allergy, type I hypersensitivity, allergic conjunctivitis, allergic rhinitis, special Shoulder dermatitis.

The pharmaceutical preparations of the invention can be made in a known manner. For example, it is manufactured by a conventional mixing, granulating, tableting, dissolving, or freeze drying process. In the manufacture of oral formulations, the mixture can be selectively milled by combining the solid adjuvant with the active compound. If necessary or necessary, after adding an appropriate amount of auxiliary agent, the mixture of particles is processed to obtain a tablet or tablet core.

Suitable excipients are, in particular, fillers, such as sugars such as lactose or sucrose, mannitol or sorbitol; cellulose preparations or calcium phosphates, such as tricalcium phosphate or calcium hydrogen phosphate; and binders, such as starch pastes, including corn starch. , wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, or polyvinylpyrrolidone. If necessary, add disintegrants such as the starch mentioned above, as well as carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate. Adjuvants, especially flow regulators and Lubricants, for example, silica, talc, stearates, such as calcium magnesium stearate, stearic acid or polyethylene glycol. If desired, the tablet core can be provided with a suitable coating that is resistant to gastric juice. For this purpose, a concentrated sugar solution can be applied. This solution may contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, a lacquer solution and a suitable organic solvent or solvent mixture. For the preparation of a gastric juice resistant coating, a suitable cellulose solution such as cellulose acetate phthalic acid or hydroxypropyl methylcellulose phthalic acid can be used. A dye or pigment can be added to the coating of the tablet or tablet core. For example, a combination for identifying or for characterizing the dosage of an active ingredient.

Based on the above compounds and pharmaceutical compositions, the invention further provides a method of treating a FLT3-mediated disease comprising administering to a subject in need thereof a compound or pharmaceutical composition of the invention.

Methods of administration include, but are not limited to, various methods of administration well known in the art, which can be determined based on the actual circumstances of the patient. These methods include, but are not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, nasal or topical routes of administration.

The invention also encompasses the use of a compound of the invention in the manufacture of a medicament for the treatment of a FLT3-mediated disease, or inhibition of FLT3.

Advantages of the invention:

1. The compound provided by the present invention is a novel pteridine derivative;

2. The compound provided by the present invention has excellent inhibitory activity against FLT3;

3. The compound provided by the invention lays a foundation for developing a targeted drug capable of inhibiting FLT3, and has great industrialization and commercialization prospects as well as market value, and the economic benefit is remarkable.

The technical solutions of the present invention are further described below in conjunction with the specific embodiments, but the following embodiments are not intended to limit the present invention, and all the application methods according to the principles and technical means of the present invention are within the scope of the present invention. The experimental methods in the following examples which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated.

Inhibitor synthesis

The invention will be further illustrated in the following examples. These examples are for illustrative purposes only and are not intended to limit the invention in any way.

Synthesis route of pteridinone core derivative

In the above preparation scheme, R ^{1 -} R ⁴ are as defined above. One skilled in the art can prepare the compounds of the present invention by using various starting compounds conventionally obtained in the art as starting materials according to actual preparation needs.

Example 1

1) Synthesis of 8-(4-aminophenyl)-2-(4-methoxy-2-methylphenyl)-7(8H)-pteridinone (F-1)

Synthesis of tert-butyl 1(4-aminophenyl)carbamate

P-phenylenediamine (890 mg, 8.23 mmol) was weighed into a 100 mL round bottom flask, 40 mL of dichloromethane was added, and the mixture was stirred in an ice bath, and di-tert-butyl dicarbonate (449 mg, 2.06 mmol) was dissolved in dichloromethane. Among them, it was added dropwise to the above reaction liquid, and then the mixture was further stirred at room temperature overnight. After completion of the reaction, the solvent was evaporated to dryness eluted eluted eluted elut elut elut elut elut elut %.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.14 (d, J = 8.0Hz, 2H), 6.65 (d, J = 8.4Hz, 2H), 6.26 (s, 1H), 1.52 (s, 9H). Mp 112-113 ° C, literature value ^[97] 112-114 ° C.

Synthesis of tert-butyl 2(4-(2-chloro-5-nitropyrimidin-4-amino)phenyl)carbamate (B-1)

Weigh 2,4-dichloro-5-nitropyrimidine (190 mg, 0.98 mmol), add 6 mL of 1,4-dioxane, place in a 25 mL round bottom flask, stir at room temperature, and take (4-aminobenzene) Tert-butyl carbamate (200 mg, 0.96 mmol), N,N-diisopropylethylamine (137 mg, 1.06 mmol) dissolved in 4 mL of 1,4-dioxane, slowly added dropwise to the above reaction mixture After the completion of the dropwise addition, stirring was continued at room temperature for about 1 hour, and the reaction was completely converted by TLC. The solvent was removed by rotary evaporation. ) tert-butyl carbamate orange solid 301 mg, yield 82%.

^{1 H NMR (400MHz, DMSO-} d 6): δ10.38 (s, 1H), 9.47 (s, 1H), 9.12 (s, 1H), 7.49 (d, J = 8.4Hz, 2H), 7.39 (d , J = 8.4 Hz, 2H), 1.49 (s, 9H). LC-MS calcd for C ₁₅ H ₁₇ ClN ₅ O ₄ [M+H] ⁺ 366.

Synthesis of tert-butyl 3(4-(2-(4-methoxy-2-methylphenylamino)-5-nitropyrimidine-4-amino)phenyl)carbamate (C-1)

(4-(2-Chloro-5-nitropyrimidin-4-amino)phenyl)carbamic acid tert-butyl ester (150 mg, 0.41 mmol), 4-methoxy-2-methylaniline (51 mg, 0.37) Methyl), N,N-diisopropylethylamine (145 mg, 1.12 mmol) was placed in a 25 mL round bottom flask, and 10 mL of 1,4-dioxane was added and stirred at room temperature overnight. After completion of the reaction, the solvent was removed by rotary evaporation. EtOAcjjjjjjjj tert-Butyl phenyl phenylamino)-5-nitropyrimidine-4-amino)phenyl)carbamate Yellow solid 148 mg, yield 85%.

^{1 H NMR (400MHz, DMSO-} d 6): δ10.22 (s, 1H), 9.86 (s, 1H), 9.33 (s, 1H), 9.06 (s, 1H), 7.60 (s, 1H), 7.36 (d, J = 8.4 Hz, 2H), 7.27-7.23 (m, 2H), 6.85 (s, 1H), 6.79 (d, J = 7.6 Hz, 1H), 3.82 (s, 3H), 2.15 (s, 3H), 1.48 (s, 9H ) .ESI calcd for _{C 23 H 27 N 6 O 5} [M + H] + 467.20, Found 467.20.

Synthesis of 4(4-(2-(4-methoxy-2-methylphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)carbamic acid tert-butyl ester (E- 1) Synthesis

Weighing (4-(2-(4-methoxy-2-methylphenylamino)-5-nitropyrimidin-4-amino)phenyl)carbamic acid tert-butyl ester (100 The mg, 0.21 mmol) was placed in a 250 mL round bottom flask, and 100 mL of methanol, 100 mL of dichloromethane, and 20 mg of palladium carbon (10% Pd) were added, and hydrogen was added thereto, and the mixture was stirred at room temperature overnight. After completion of the reaction, the mixture was filtered under suction, and the filtrate was evaporated to dryness to give the crude product 88 mg.

The product obtained in the previous step was placed in a 50 mL round bottom flask, 0.6 mL of glacial acetic acid, 10 mL of absolute ethanol was added, and then ethyl glyoxylate (50% toluene solution) (0.044 mL, 0.22 mmol) was added and heated to reflux. Stir overnight. After the completion of the reaction, a solid precipitated, suction filtration, and the filter cake was washed with ethanol and dried. The crude product was purified by silica gel column chromatography (dichloromethane/methanol=50:1, v/v) to afford (4-(2-(4-methoxy-2-methylphenylamino)-7- Oxo-8(7H)-pteridinylphenyl)carbamic acid tert-butyl ester 61 mg (yield: 64%).

^{1 H NMR (400MHz, DMSO-} d 6): δ9.55 (s, 1H), 9.17 (s, 1H), 8.76 (s, 1H), 7.98 (s, 1H), 7.56 (d, J = 8.8Hz , 2H), 7.22 (d, J = 7.6 Hz, 2H), 7.16 (s, 1H), 6.69 (s, 1H), 6.54 (s, 1H), 3.70 (s, 3H), 2.11 (s, 3H) , 1.51 (s, 9H) .ESI calculated for _{C 25 H 27 N 6 O 4} [M + H] + 475.20, Found 475.20.

Synthesis of 58-(4-Aminophenyl)-2-(4-methoxy-2-methylphenyl)-7(8H)-pteridinone (F-1)

(4-(2-(4-Methoxy-2-methylphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)carbamic acid tert-butyl ester (50 mg, 0.11) Methyl) was placed in a 25 mL round bottom flask, 6 mL dichloromethane was added, stirred at 0 ° C, and 1.5 mL of trifluoroacetic acid was added. Stirring was continued at 0 ° C for 1 hour and then at room temperature for 2 hours. After completion of the reaction, it was neutralized by adding a saturated sodium hydrogencarbonate solution until the solution was alkaline, and extracted with dichloromethane. The organic phase was washed with deionized water, dried over anhydrous sodium sulfate and evaporated. The crude product was purified by silica gel column chromatography (dichloromethane/methanol=30:1, v/v) to give 8-(4-aminophenyl)-2-(4-methoxy-2-methylbenzene Base 7-(8H)-pteridinone yellow solid 32 mg, yield 82%, mp 149.1 - 149.3.

^{1 H NMR (400MHz, DMSO-} d 6): δ9.13 (s, 1H), 8.72 (s, 1H), 7.95 (s, 1H), 7.21 (s, 1H), 6.94 (d, J = 8.0Hz , 2H), 6.72 (s, 1H), 6.65 (d, J = 8.4 Hz, 2H), 6.60 (s, 1H), 5.33 (br, 2H), 3.72 (s, 3H), 2.13 (s, 3H) .HRMS (ESI) calcd for _{C 20 H 19 N 6 O 2} [M + H] + 375.1569, Found 375.1573. HPLC purity: 96.0%, retention time = 11.68 min.

2) Synthesis of 8-(4-aminophenyl)-2-(3-chloro-4-methoxyphenyl)-7(8H)-pteridinone (F-2)

The synthesis method was carried out with reference to F-1 to give a yellow solid, yield 75%, mp > 300 °C.

^{1 H NMR (400MHz, DMSO-} d 6): δ10.13 (s, 1H), 8.84 (s, 1H), 8.03 (s, 1H), 7.60 (s, 1H), 7.38 (s, 1H), 6.98 (d, J = 8.4 Hz, 2H), 6.87 (s, 1H), 6.72 (d, J = 8.4 Hz, 2H), 5.39 (br, 2H), 3.79 (s, 3H). HRMS (ESI) C ₁₉ H ₁₆ ClN ₆ O ₂ [M+H] ⁺ 395.1023. HPLC purity: 97.0%, retention time=11.55min.

3) Synthesis of 8-(4-aminophenyl)-2-(3-fluoro-4-methoxyphenyl)-7(8H)-pteridinone (F-3)

The synthesis method was carried out with reference to F-1 to give a yellow solid, yield 73%, mp >

^{1 H NMR (400MHz, DMSO-} d 6): δ10.14 (s, 1H), 8.85 (s, 1H), 8.03 (s, 1H), 7.42 (s, 1H), 7.19 (s, 1H), 6.98 (d, J = 8.4 Hz, 2H), 6.89 (s, 1H), 6.72 (d, J = 8.4 Hz, 2H), 5.38 (br, 2H), 3.77 (s, 3H). HRMS (ESI) C ₁₉ H ₁₆ FN ₆ O ₂ [M+H] ⁺ 379.1 319. HPLC purity: 96.3%, retention time = 11.68 min.

4) Synthesis of N-(4-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acetamide (E-4)

The synthesis method was carried out with reference to E-1 to give a yellow solid, yield 62%, mp >

^{1 H NMR (400MHz, DMSO-} d 6): δ10.23 (s, 1H), 10.10 (s, 1H), 8.85 (s, 1H), 8.04 (s, 1H), 7.78 (d, J = 7.2Hz , 2H), 7.34 (d, J = 7.6 Hz, 4H), 6.61 (s, 2H), 3.67 (s, 3H), 2.13 (s, 3H). HRMS (ESI) calculated C ₂₁ H ₁₉ N ₆ O ₃ [M+H] ⁺ 403.1519, experimental value 403.1500. HPLC purity: 97.5%, retention time=10.54min.

5) Synthesis of N-(4-(2-(4-methylphenyl)amino)-7-oxo-8(7H)pteridinyl)phenyl)methylsulfonamide (G-5)

8-(4-Aminophenyl)-2-(4-methoxyphenyl)-7(8H)-pteridinone (100 mg, 0.277 mmol), triethylamine (0.232 mL, 1.662 mmol) In a 25 mL round bottom flask, 15 mL of anhydrous dichloromethane was added and stirred under ice bath. Methanesulfonyl chloride (0.172 mL, 2.216 mmol) was weighed and dissolved in 2 mL of dichloromethane, and the mixture was slowly added dropwise to the reaction mixture. After the dropwise addition, stirring was continued for 1 hour in an ice bath and stirred at room temperature overnight. TLC tracks to complete conversion of the feedstock. The solvent was evaporated to dryness, and the mixture was evaporated to drynessnessness The crude product was purified by silica gel column chromatography (dichlorodiethyl acetate / EtOAc / EtOAc / 7-Oxo-8(7H)pteridinyl)phenyl)methylsulfonamide 54 mg,yield 47%, mp > 300.

^{1 H NMR (400MHz, DMSO-} d 6): δ10.11 (br, 1H), 8.84 (s, 1H), 8.03 (s, 1H), 7.40 (s, 4H), 7.31 (br, 2H), 6.66 (br, 2H), 3.69 ( s, 3H), 3.12 (s, 3H) .HRMS (ESI) calcd for _{C 20 H 19 N 6 O 4} S [M + H] + 439.1189, Found 439.1187. HPLC purity: 95.4%, retention time = 11.70 min.

6) 8-(4-Aminophenyl)-2-((3-chloro-4-(4-methyl-1-piperazinyl)phenyl)amino)-7(8H)pteridone (F- 6) Synthesis

The synthesis method was carried out with reference to F-1 to give a yellow solid (yield: 62%).

^{1 H NMR (400MHz, DMSO-} d 6): δ10.13 (br, 1H), 8.85 (s, 1H), 8.03 (s, 1H), 7.60 (s, 1H), 7.37 (d, J = 7.6Hz , 1H), 6.97 (d, J = 8.4 Hz, 2H), 6.89 (d, J = 7.6 Hz, 1H), 6.71 (d, J = 8.4 Hz, 2H), 5.38 (s, 2H), 2.89 (br , 4H), 2.47 (br, 4H), 2.34 (s, 3H) .HRMS (ESI) calcd for _{C 23 H 24 ClN 8 O [} M + H] + 463.1762, Found 463.1710. HPLC purity: 98.7%, retention time = 9.64min.

7) 8-(4-Aminophenyl)-2-((3-fluoro-4-(4-methyl-1-piperazinyl)phenyl)amino)-7(8H)pteridone (F- 7) Synthesis

The method of synthesis was carried out with reference to F-1 to give a yellow solid (yield: 64%, mp.

^{1 H NMR (400MHz, DMSO-} d 6): δ10.13 (br, 1H), 8.84 (s, 1H), 8.02 (s, 1H), 7.32 (d, J = 12.4Hz, 1H), 7.19 (d , J = 7.2 Hz, 1H), 6.97 (d, J = 8.4 Hz, 2H), 6.77 (br, 1H), 6.72 (d, J = 8.4 Hz, 2H), 5.39 (s, 2H), 2.92 - 2.90 (m, 4H), 2.45 ( br, 4H), 2.22 (s, 3H) .HRMS (ESI) calcd for _{C 23 H 24 FN 8 O [} m + H] + 447.2057, Found 447.2057. HPLC purity: 96.9%, retention time = 9.12 min.

8) 8-(4-Aminophenyl)-2-((3-methyl-4-(4-methyl-1-piperazinyl)phenyl)amino)-7(8H)pteridone (F -8) Synthesis

The synthesis method was carried out with reference to F-1 to give a yellow solid, yield 74%, mp 255.6 -256.3.

^{1 H NMR (400MHz, DMSO-} d 6): δ9.94 (br, 1H), 8.80 (s, 1H), 7.99 (s, 1H), 7.36 (br, 1H), 7.18 (d, J = 6.4Hz , 1H), 6.97 (d, J = 8.4 Hz, 2H), 6.79 (d, J = 8.4 Hz, 1H), 6.71 (d, J = 8.4 Hz, 2H), 5.37 (s, 2H), 2.76-2.74 (m, 4H), 2.47 (br, 4H), 2.24 (s, 3H), 2.06 (s, 3H). HRMS (ESI) calc. C ₂₄ H ₂₇ N ₈ O [M+H] ⁺ 443.2308, experimental value 443.2301. HPLC purity: 99.2%, retention time=9.44min.

9) 8-(4-Aminophenyl)-2-((3-methoxy-4-(4-methyl-1-piperazinyl)phenyl)amino)-7(8H)pteridone ( Synthesis of F-9)

The synthesis method was carried out with reference to F-1 to give an orange-yellow solid, yield 63%, mp 147.4 - 147.9 °.

^{1 H NMR (400MHz, DMSO-} d 6): δ9.94 (br, 1H), 8.81 (s, 1H), 7.99 (s, 1H), 7.09 (s, 2H), 6.97 (d, J = 8.4Hz , 2H), 6.70 (d, J = 8.4 Hz, 2H), 6.60 (br, 1H), 5.41 (s, 2H), 3.56 (s, 3H), 2.87 (br, 4H), 2.43 (br, 4H) , 2.21 (s, 3H) .HRMS (ESI) calcd for _{C 24 H 27 N 8 O 2} [M + H] + 459.2257, Found 459.2256. HPLC purity: 98.0%, retention time = 8.75min.

10) 8-(4-Aminophenyl)-2-((3-methyl-4-(1-methyl-4-oxopiperidyl)phenyl)amino)-7(8H)pteridone Synthesis of (F-10)

The synthesis method is referred to as F-1 to obtain a yellow solid, the yield is 52%, mp 228.9-229.4 ° C.

^{1 H NMR (400MHz, DMSO-} d 6): δ9.97 (br, 1H), 8.81 (s, 1H), 7.99 (s, 1H), 7.36 (d, J = 2.0Hz, 1H), 7.16 (br , 1H), 6.97 (d, J = 8.8 Hz, 2H), 6.70 (d, J = 8.4 Hz, 3H), 5.39 (s, 2H), 4.37 (s, 1H), 2.92 (s, 2H), 2.75 (s, 2H), 2.01 (s, 6H), 1.81-1.71 (m, 2H), 1.34-1.19 (m, 1H). HRMS (ESI) calculated C ₂₅ H ₂₈ N ₇ O ₂ [M+H] ⁺ 458.2304, experimental value 458.2289. HPLC purity: 99.3%, retention time=10.29min.

11) N-(4-(2-(4-(4-methyl-1-piperazinyl)amino)-7-oxo-8(7H)-pteridinone)phenyl)acetamide (E- 11) Synthesis

The synthesis method was carried out according to E-1 to give an orange-red solid, yield 58%, mp 288.0-288.7 °C.

^{1 H NMR (400MHz, DMSO-} d 6): δ10.44 (s, 1H), 10.05 (s, 1H), 8.82 (s, 1H), 8.02 (s, 1H), 7.80 (d, J = 8.0Hz , 2H), 7.32 (d, J = 8.4 Hz, 2H), 7.25 (br, 2H), 6.64 (br, 2H), 3.24 (br, 4H), 3.05 (br, 4H), 2.64 (s, 3H) , 2.15 (s, 3H) .HRMS (ESI) calcd for _{C 25 H 27 N 8 O 2} [M + H] + 471.2257, Found 471.2213. HPLC purity: 97.3%, retention time=8.17min.

12) N-(4-(2-(4-(4-methyl-1-piperazinyl)phenyl)amino)-7-oxo-8(7H)pteridinyl)phenyl)methyl Synthesis of Sulfonamide (G-12)

The synthesis method was carried out with reference to F-1 to give an orange-red solid, yield 86%.

^{1 H NMR (400MHz, DMSO-} d 6): δ9.98 (s, 1H), 8.78 (s, 1H), 7.98 (s, 1H), 7.32 (d, J = 6.4Hz, 2H), 6.97 (d , J = 8.8 Hz, 2H), 6.74 (d, J = 8.0 Hz, 2H), 6.67 (br, 2H), 3.04-3.02 (m, 4H), 2.45 - 2.43 (m, 4H), 2.22 (s, 3H) .HRMS (ESI) calcd for _{C 23 H 25 N 8 O [} M + H] + 429.2151, Found 429.2153.

1N-(4-(2-(4-(4-methyl-1-piperazinyl)phenyl)amino)-7-oxo-8(7H)pteridinyl)phenyl)methylsulfonamide Synthesis of (G-12)

The synthesis method was carried out with reference to G-5 to give a yellow solid, yield 50%, mp > 300 °C.

^{1 H NMR (400MHz, DMSO-} d 6): δ10.02 (s, 1H), 8.82 (s, 1H), 8.02 (s, 1H), 7.42-7.37 (m, 4H), 7.26 (br, 2H) , 6.66 (br, 2H), 3.13 (s, 3H), 3.07 (br, 4H), 2.54 (br, 4H), 2.29 (s, 3H). HRMS (ESI) calcd for _{C 24 H 27 N 8 O 3} S [M + H] + 507.1927, Found 507.1926. HPLC purity: 99.0%, retention time = 8.53min.

13) Synthesis of 8-(4-aminophenyl)-2-((4-(4-methyl-1-piperazinyl)benzyl)amino)-7(8H)pteridone (F-13)

The synthesis method was carried out with reference to F-1 to give a yellow solid, yield 65%, mp 216.2 - 216.8.

^{1 H NMR (400MHz, DMSO-} d 6): δ9.99 (s, 1H), 8.82 (s, 1H), 8.01 (s, 1H), 7.55 (d, J = 8.0Hz, 2H), 7.32 (d , J=8.4 Hz, 2H), 7.20 (s, 2H), 6.59 (s, 2H), 3.86 (s, 2H), 3.00-2.99 (m, 4H), 2.44-2.41 (m, 4H), 2.21 ( s, 3H) .HRMS (ESI) calcd for _{C 24 H 27 N 8 O [} M + H] + 443.2308, Found 443.2304. HPLC purity: 97.4%, retention time = 6.92 min.

14) 2-((4-(4-Methyl-1-piperazinyl)phenyl)amino)-8-(4-(1-piperazinyl)phenyl)-7(8H)pteridone ( Synthesis of F-14)

The synthesis method was carried out with reference to F-1 to give a yellow solid, yield 72%, mp 276.6-277.2 °C.

^{1 H NMR (400MHz, DMSO-} d 6): δ10.01 (s, 1H), 8.81 (s, 1H), 8.00 (s, 1H), 7.26 (d, J = 6.8Hz, 2H), 7.20 (d , J = 8.8 Hz, 2H), 7.12 (d, J = 8.8 Hz, 2H), 6.58 (s, 2H), 3.19 - 3.17 (m, 4H), 3.02-3.00 (m, 4H), 2.90 - 2.88 ( m, 4H), 2.45-2.43 (m, 4H), 2.22 (s, 3H). HRMS (ESI) calcd. C ₂₇ H ₃₂ N ₉ O [M+H] ⁺ 498.2730. HPLC purity: 98.6%, retention time = 7.86min.

15) 8-(3-Aminophenyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)pteridin-7(8H)-one. (F-15)

The synthesis method is referred to as F-1, and a yellow solid is obtained in a yield of 70%, mp 262.5-262.9 ° C.

^{1 H NMR (400MHz, DMSO-} d 6): δ10.01 (s, 1H), 8.81 (s, 1H), 7.99 (s, 1H), 7.35 (d, J = 7.2Hz, 2H), 7.22 (t , J = 8.0 Hz, 1H), 6.75 (d, J = 7.2 Hz, 1H), 6.66 (br, 2H), 6.52 (br, 1H), 6.48 (d, J = 8.0 Hz, 1H), 5.35 (s , 2H), 3.02 (br, 4H), 2.46-2.44 (m, 4H), 2.23 (s, 3H). HPLC purity: 99.5%, retention time = 8.57min. HRMS (ESI) (m/z): M+H] ⁺ calcd for C ₂₃ H ₂₅ N ₈ O, 429.2151; found, 429.2151.

16) 8-(4-N,N-Dimethylaminophenyl)-2-((3-methyl-4-(4-methylpiperazin-1-yl)phenyl)amino)pteridin-7 (8H)-ketone. (F-16)

The raw material 1 was placed in a 250 ml round bottom flask, dissolved in DCM and MeOH, 100 mg of palladium carbon (10% Pd) was weighed, hydrogen gas was introduced, and the reaction was carried out for about 10 hours at room temperature, followed by suction filtration, and the filtrate was evaporated under reduced pressure to give a solvent. It was used directly in the next step without purification. Weigh 2 (0.3 g, 0.69 mmol) in a 50 ml round bottom flask, add an appropriate amount of ethanol to dissolve, add 0.21 ml (1.04 mmol) of ethyl glyoxylate (50% toluene solution) and 0.6 ml of glacial acetic acid with stirring, and heat up to The mixture was refluxed, and the mixture was applied to EtOAc (EtOAc).

^{1 H NMR (400MHz, DMSO-} d 6, ppm) δ10.02 (br, 1H), 8.83 (s, 1H), 8.01 (s, 1H), 7.28 (s, 1H), 7.15 (d, J = 8.8 Hz, 3H), 6.87 (d, J = 8.8 Hz, 2H), 6.69-6.67 (m, 1H), 3.01 (s, 6H), 2.71 (m, 4H), 2.44 (m, 4H), 2.22 (s) , 3H), 1.96 (s, 3H). MS (ESI) (m/z): [M+H] ⁺ calcd for C ₂₆ H ₃₀ N ₈ O, 470.25; found, 470.21.

Example 2

The synthesis procedure of Compound 1 is as follows, and the synthesis of Compound 2-31 is carried out with reference to the synthesis of Compound 1:

Synthesis of (S)-3-((2-chloro-5-nitropyrimidine) 4-amino)pyrrolidine-1-carboxylic acid tert-butyl ester

2,4-Dichloro-5-nitropyrimidine (3.23 g, 16.28 mmol) was weighed into a 250 mL low temperature reaction flask, dissolved in 80 mL of dichloromethane, stirred at -70 ° C, and taken (S)-1 - tert-Butoxycarbonyl-3-aminopyrrolidine (3.03 g, 16.28 mmol), N,N-diisopropylethylamine (2.52 g, 19.53 mmol) dissolved in dichloromethane (20 mL) After the completion of the dropwise addition, stirring was continued at -70 ° C for 0.5 hour, and TLC was traced to complete conversion of the starting material. The solvent was removed by rotary evaporation. EtOAc EtOAcjjjjjj

^{1 H NMR (400MHz, DMSO-} d 6): δ9.07 (s, 1H), 8.41 (d, J = 6.4Hz, 1H), 4.87-4.84 (m, 1H), 3.81 (dd, J = 11.6Hz , 6.4 Hz, 1H), 3.54 (t, J = 6.4 Hz, 1H), 3.36 (br, 1H), 2.40 - 2.31 (m, 1H), 2.06-1.98 (m, 1H), 1.48 (s, 9H) .

Synthesis of 1-methyl-4-(2-methyl-4-nitrophenyl)piperazine

3-Methyl-4-fluoro-1-nitrobenzene (2.67 g, 17.21 mmol) and N-methylpiperazine (1.72 g, 17.21 mmol) were weighed into a 50 ml round bottom flask, dissolved in DMSO, and added to K. ₂ CO ₃ 4.76g, heated and stirred at 90 °C for 9h, TLC tracking to complete conversion of the raw materials, removal of K ₂ CO _{3 by} suction filtration, washing with ethyl acetate, taking the filtrate and diluting with a small amount of ethyl acetate, extracting with saturated sodium chloride, organic layer The solvent was evaporated to dryness afforded EtOAcqqqqqqqq

^{1 H NMR (400MHz, DMSO-} d 6): δ7.88 (d, J = 4.8Hz, 1H), 6.60 (dd, J = 9.6Hz, 2.4Hz, 1H), 6.53 (s, 1H), 3.91 ( s, 3H), 3.44 (br, 4H), 2.42 (br, 4H), 2.22 (s, 3H).

Synthesis of 3-methyl-4-(4-methylpiperazinyl)aniline

Weigh S2 (1.96g, 8.33mmol) in a 250ml round bottom flask, add ethyl acetate / methanol (1:1) to dissolve, add 100mg 10% Pd / C, add hydrogen, stir at room temperature for 4h, TLC tracking until the raw materials are completely After conversion, the reaction solution was suction filtered, and then evaporated and evaporated. The product was used in the next step without purification.

(S)-3-((2-((3-methyl-4-(4-methyl-1-piperazinyl)phenyl)amino)-5-nitro-4-pyrimidinyl)amino)pyrrole Synthesis of alkyl-1-tert-butylformate

S1 (2.84 g, 8.26 mmol), S2 (1.70 g, 8.26 mmol), DIPEA (2.13 g, 16.52 mmol) was weighed in a 250 ml round bottom flask, dissolved in THF and stirred overnight under argon. The TLC was traced to complete conversion of the starting material, and the solvent was removed by rotary evaporation, diluted with water and extracted with ethyl acetate (100 mL×2). The organic phase was washed with saturated ammonium chloride solution (50 mL×2), saturated sodium chloride solution, and then added. Dry over anhydrous sodium sulfate. The crude product was purified by silica gel column chromatography eluting elut elut elut elut elut elut

^{1 H NMR (400MHz, DMSO-} d 6): δ10.35 (s, 1H), 8.95 (s, 1H), 8.57 (s, 1H), 7.69 (s, 1H), 7.44 (d, J = 7.6Hz , 1H), 6.98 (d, J = 8.8 Hz, 1H), 4.65 - 4.62 (m, 1H), 4.12 (s, 0.4H), 3.72 (dd, J = 10.8 Hz, 6.8 Hz), 3.57-3.27 ( m, 4.31H), 3.18 (s, 1H), 2.80 (br, 4H), 2.45 (br, 4H), 2.22 (s, 3H), 2.21 (s, 3H), 1.42 (s, 9H).

(S)-3-((5-Amino-2-((3-methyl-4-(4-methyl-1-piperazinylphenyl)amino)4-pyrimidinyl)amino)pyrrolidinyl) Tert-butyl 1-carboxylic acid

S4 (3.45 g, 6.73 mmol) was weighed in a 250 ml round bottom flask, dissolved in absolute ethanol, 100 mg Pd/C (10%) was added, hydrogen gas was added, and the mixture was stirred at room temperature for 8 h. After the reaction, suction filtration was performed. The filtrate was spun dry and the product was not purified. Used directly in the next step.

(S)-3-(6-isopropyl-2-((3-methyl-4-(4-methyl-1-piperazinyl)phenyl)amino)-7-oxo-8 (7H) )-pteridinylpyrrolidin-1-carboxylic acid tert-butyl ester

The product of the previous step S5 (1.61 g, 3.33 mmol) was taken in a 25 ml round bottom flask, 10 ml of absolute ethanol, 0.6 ml of glacial acetic acid was added, and ethyl dimethylacetate (721 mg, 5.00 mmol) was added and heated under argon atmosphere. Return and stir overnight. The TLC was traced to complete conversion of the starting material, and the solvent was removed by rotary evaporation. The mixture was diluted with water and extracted with dichloromethane (100 mL×2). The organic phase was washed with saturated sodium hydrogen carbonate solution (50 mL×2) and saturated sodium chloride solution, and then added. Dry with sodium sulfate. The crude product was subjected to EtOAcjjjjjjjlilililililililililililili Lc-ms

(S)-6-isopropyl-2-((3-methyl-4-(4-methyl-1-piperazinyl)phenyl)amino)-8-(3-pyrrolidinyl)-7 Synthesis of (8H) pteridinone

Weighed S6 (1.20g, 2.13mmol) in a 25ml round bottom flask, dissolved in dichloromethane, cooled in an ice bath, and then added 4ml of trifluoroacetic acid, and reacted at room temperature for 3h, TLC tracking to complete conversion of the raw materials, reaction solution Pour into a beaker, dilute with dichloromethane, and add sodium hydroxide solution to the solution while stirring. The upper layer has a pH of about 9, dilute with dichloromethane, extract with saturated sodium chloride solution, and dry over anhydrous sodium sulfate. The crude product was subjected to EtOAcjjjjjjjjjjj

^{1 H NMR (400MHz, DMSO-} d 6): δ9.85 (s, 1H), 8.74 (s, 1H), 7.51 (s, 1H), 7.43 (d, J = 8.4Hz, 1H), 7.02 (d , J=8.4 Hz, 1H), 5.85-5.77 (m, 1H), 3.42-3.36 (m, 1H), 2.96-2.91 (m, 1H), 2.82 (br, 4 Hz), 2.78-2.73 (m, 1H) ), 2.47 (br, 4H), 2.24 (s, 3H), 2.23 (s, 3H), 2.17-2.09 (m, 1H), 1.99-1.91 (m, 1H), 1.19 (d, J = 6.8 Hz, 6H).

Example 3

Biological activity test section

Kinase test method

Tested by Mobility shift assay, kinase buffer (50 mM HEPES, pH 7.5, 0.01% Brij-35, 10 mM MgCl ₂ , 2 mM DTT), stop buffer (100 mM HEPES, pH 7.5, 0.015% Brij-35, 0.2) %Coating Reagent#3, 50 mM EDTA). The compound was diluted to the desired concentration with 100% DMSO, and the formulated compound was transferred to a 96-well plate at a concentration ranging from 20 μl to 60 μl and equilibrated to 10 concentrations with a concentration gradient. A 100 μl concentration of 100% DMSO was used as a blank control. Thereafter, 5 μl of the drug in the 96-well plate was transferred to a 384-well plate and the test was repeated. The kinase was cultured in a specific manner at 28 °C. Finally, the IC ₅₀ value was calculated from the standard curve.

Cell test method

Cell culture: Cell lines were obtained from the American Type Culture Collection. The cells were cultured in RPMI-1640 or MEM medium containing 10% fetal bovine serum in 5% CO ₂ at 37 °C. MV4-11 cells were cultured in IMDM medium.

Cell viability assay method: The activity test of suspension cells is mainly determined by the MTT test method, and the test of adherent cells is mainly the SRB test method. The suspension cells were cultured in 96-well plates of 3-8 x 10 ³ cells per well, 100 μL of the medium was added to each well, and then 10 μL of the compound (in triplicate) was added, and cultured at 37 ° C for 72 hours. After the cells were cultured, 20 μL of 5 mg/mL MTT reagent was added to each well plate for further 4 hours, and 50 μL of three mixed solvents (5% isopropanol-10% SDS-12 mmol/L HCl) were added to dissolve the oxidation product. . The OD value was measured at 570 nm using a multifunction detector Synergy2. The adherent cells were cultured in 96-well plates of 2-7 x 10 ³ cells per well for 24 h, and the same equivalent of the compound was also added to the medium. After 72 h of culture, the culture solution was removed, and the strain was fixed with TCA for 1 h. The dried well plates were stained with sulforhodamine B and then 150 μL of 10 mM Tris-HCl was added. Finally, the light absorption of the lysed cells was detected at 560 nm using a Synergy 2 differential spectrophotometer. All experiments were performed in triplicate.

Immunoprecipitation and Western blotting experiments: Cells were incubated with different concentrations of compounds for 2 h, then the cells were further solubilized with RIPA lysis buffer, then centrifuged at 4 ° C, and the supernatant was taken for relevant experiments. The protein was separated by SDS-PAGE and then transferred to a nitrocellulose membrane for antibody detection. The primary antibodies act on FLT3, AKT, p-AKT, STAT5, p-STAT5 and p44/42 MAPK, and anti-GAPDH is cultured at the recommended concentrations. The visualized signal was used to detect the elevated ECL matrix using the Tanon 5200 capture system after HRP copolymerization with the antibody. The cell lysate was immunoprecipitated with FLT3 antibody and protein A gel, and the cooked sample was electrophoretically polarized with 6% gel, and then labeled with 4G10 antibody, and the FLT3 phosphorylation level was detected by this method.

Flow cytometry: Cells were stained with Annexin V and propidium iodide for the detection of apoptotic activity of the compounds. MV4-11 cells were cultured in a 6-well plate at a density of 1 × 10 ⁵ cells/mL, and then cultured at different concentrations for 48 hours. The cells were divided and washed twice with cold PBS buffer solution. The sample was resuspended in binding buffer and incubated with annexin V and propidium iodide under the protection of light. The cell cycle assay was performed according to the PI/Rnase staining solvent product specification. The dividing cells were fixed in 70% ethanol for 1 h. All cells were then stained with propidium iodide for 30 min under non-illuminated room temperature conditions. Apoptosis and cell cycle were detected by FACS flow cytometry.

The test results of the compounds (tests with AC220 and MLN518 as positive control drugs) are shown in the following table, and the results of the cell test are shown in the attached drawings.

Table 1. Kinase and cell test results for compounds

MV4-11 xenograft model test

The inventors further tested the inhibitory activity of the compounds of the present invention on tumors in the MV4-11 xenograft model.

Results As shown in Figures 2-3, the effect of the compounds of the invention on tumor volume and weight was similar to that of the positive control AC220, further confirming the potential of the compounds of the invention in inhibiting tumors.

discuss:

After extensive and in-depth research, the inventors designed and synthesized a series of pteridinone derivatives that have not been reported in the literature. The obtained compounds were tested at the molecular and cellular levels to obtain a batch of compounds capable of inhibiting FLT3. . The IC ₅₀ value of the inhibitory activity of these compounds on FLT3 reached nM level; the IC ₅₀ value of inhibitory activity against related cells reached the μM level. Among them, a plurality of compounds, such as F-8 and F-16, have an inhibitory activity against FLT3 with an IC ₅₀ value of less than 10 nM, which is far superior to the positive control MLN 518. Further tumor model tests have shown that the compounds of the invention also have excellent tumor suppressor activity in vivo. Thus, the non-compounds of the invention may be the target drug for inhibiting FLT3, or the basis for further modification to give compounds that are more active and/or less toxic.

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the appended claims.

Claims (10)

1. A compound having the structure of Formula I or a pharmaceutically acceptable salt thereof:

   

   In the formula,

   X is selected from: N, CH;

   Y is selected from the group consisting of: O, S, and Se;

   A is selected from various substituted benzene rings, nitrogen-containing five-membered rings, nitrogen-containing six-membered rings, for example

   

   R ^{1 is} independently selected from the group consisting of hydrogen, halogen, C ₁ -C ₈ alkyl, C ₃ -C ₆ cycloalkyl, fluorine-containing C ₁ -C ₈ alkyl, optionally substituted phenyl;

   R ^{2 is} selected from the group consisting of hydrogen, optionally substituted C ₁ -C ₆ alkyl, halogen, optionally substituted C ₁ -C ₆ alkoxy, amino, -NR _a R _b , carboxyl, optionally substituted alkoxy Formyl, optionally substituted N-alkylpiperazin-1-yl, optionally substituted morpholin-4-yl, optionally substituted piperidin-1-yl, optionally substituted pyrrol-1-yl, Optionally substituted pyrrolidin-1-yl, optionally substituted aryl or optionally substituted arylmethyl, optionally substituted piperidinyloxy, optionally substituted N,N',N'-three Alkyl ethylenediamine;

   R _a and R _b are each independently selected from the group consisting of: hydrogen, C ₁ -C ₆ alkyl and alkenyl;

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

   R ^{3 is} each independently selected from the group consisting of hydrogen, halogen, hydroxy, amino, -NR _c R _d , C ₁ -C ₂ acylamino, optionally substituted C ₁ -C ₆ alkyl, C ₁ -C ₃ sulfonylamino, CN, carbamoyl, carboxyl, C1-C6 alkoxycarbonyl, optionally substituted phenyl, optionally substituted piperazinyl, optionally substituted morpholin-4-yl, optionally substituted pyrrol-1- Base, optionally substituted pyrrolidin-1-yl;

   R _c and R _d are each independently selected from the group consisting of: hydrogen, C ₁ -C ₆ alkyl and alkenyl;

   m is 0, 1, 2, 3 or 4;

   Wherein, when both n and m are 1, and R ² is a C ₁ -C ₆ alkoxy group, R ^{3 is} not an amino group.
2. The compound of claim 1 wherein said compound has the structure of formula II:

   

   In the formula,

   A is selected from various substituted benzene rings;

   R ¹ is hydrogen;

   R ^{2 is} selected from the group consisting of hydrogen, optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₁ -C ₆ alkoxy, halogen, amino, -NR _a R _b , optionally substituted piperazinyl, optionally Substituted N-alkylpiperazin-1-yl, optionally substituted piperidinyloxy;

   R _a and R _b are each independently selected from the group consisting of: hydrogen, C1-C6 alkyl and alkenyl;

   n is 1, 2, or 3;

   R ^{3 is} each independently selected from the group consisting of: hydrogen, halogen, C1-C2 acylamino, hydroxy, amino, -NR _c R _d , methanesulfonylamino, optionally substituted C ₁ -C ₆ alkyl, optionally substituted piperazine base;

   R _c and R _d are each independently selected from the group consisting of: hydrogen, C ₁ -C ₃ alkyl and alkenyl;

   m is 1, 2, or 3.
3. The compound of claim 2 wherein said compound has the structure of formula III:

   

   In the formula,

   R ^{4 is} selected from the group consisting of hydrogen, halogen, optionally substituted C ₁ -C ₆ alkoxy, optionally substituted C ₁ -C ₆ alkyl;

   R ^{5 is} selected from the group consisting of: an optionally substituted N-alkylpiperazin-1-yl, optionally substituted piperidinyloxy;

   R ^{6 is} selected from the group consisting of: an amino group, an optionally substituted piperazinyl group, -NR _e R _f ;

   R _e and R _f are each independently selected from the group consisting of hydrogen, C ₁ -C ₃ alkyl and alkenyl.
4. The compound of claim 3, wherein

   In the formula,

   R ^{4 is} selected from an optionally substituted C ₁ -C ₆ alkyl group, preferably a C ₁ -C ₃ alkyl group;

   R ^{5 is} selected from the group consisting of an optionally substituted N-alkylpiperazin-1-yl;

   R ^{6 is} selected from amino, or -NR _e R _f ;

   R _e and R _f are each independently selected from the group consisting of hydrogen and C ₁ -C ₃ alkyl.
5. a compound selected from the group consisting of: or a pharmaceutically acceptable salt thereof:

   

   

   

   
6. A pharmaceutical composition comprising the compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.
7. Use of a compound according to any one of claims 1 to 5 for the manufacture of a medicament for the treatment of a FLT3-mediated disease, or for the inhibition of FLT3.
8. The use according to claim 7, wherein the FLT3-mediated disease is a blood disease such as a myeloproliferative disease, a cancer, an immune disease, and a skin disease such as psoriasis and atopic dermatitis.
9. The use according to claim 8, wherein the cancer is selected from the group consisting of acute myeloid leukemia (AML), mixed line leukemia (MLL), T-cell acute leukemia (T-ALL), and B-cell acute leukemia (B). -ALL), chronic myelomonocytic leukemia (CMML), chronic lymphocytic leukemia, chronic myeloid leukemia, chronic neutrophilic leukemia.
10. The use according to claim 7, wherein the immune disease is selected from the group consisting of arthritis, lupus, inflammatory bowel disease, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, and Stil Disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Odd thyroiditis, Graves' disease, rheumatoid arthritis syndrome, multiple sclerosis, infectious neuronitis, acute transmission Encephalomyelitis, Addison's disease, aplastic anemia, autoimmune hepatitis, optic neuritis, silvery disease, graft versus host disease, transplantation, blood transfusion allergy, allergy, type I hypersensitivity, allergic conjunctiva Inflammation, allergic rhinitis, atopic dermatitis.

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