WO2018004213A1 - Novel compound having smo inhibitory activity and composition comprising same as active ingredient for preventing or treating cancer - Google Patents

Abstract

The present invention relates to a novel compound having an SMO inhibitory activity, wherein the novel compound exhibits an excellent anticancer effect through abilities of isoindolinone and quinazolinone derivative compounds to modulate the hedgehog signaling pathway and inhibit SMO activity, and thus can prevent, alleviate, or treat a cancer disease.

Description

Novel compounds having SMO inhibitory activity and compositions for preventing or treating cancer comprising the same as active ingredients

The present invention provides novel isoindolinone or quinazolinone derivative chemicals that act as SMO (smoothened) inhibitors to inhibit cancer caused by Hedgehog signaling hyperactivity. It relates to a pharmaceutical composition for the prevention or treatment of cancer diseases, including as an active ingredient.

Hedgehog (Hh) is a protein that plays an important role in embryonic development and regulates proliferation, migration and differentiation in various organs including dorsal and embryonic axis formation, nervous system, musculoskeletal system, etc. Works. Hedgehog in the adult is involved in the maintenance of stem cells, abnormal abnormal activity in the adult tissue of the hedgehog causes a variety of cancers, such as basal cell carcinoma, brain cancer.

Hedgehog signaling abnormalities in the human developmental stage cause stillbirth or organ development disorders because they affect a wide range of processes from the beginning of fetal development to the end organ formation process. After development, hedgehog signaling activity is maintained only in some specific tissues, such as adult stem cells, in adults and is reduced in most tissue somatic cells. On the contrary, excessive signaling activity caused by mutations of hedgehog signaling genes (Ptch, Smo, Sufu, Gli, etc.) at the somatic stage promotes cell proliferation and causes cancer.

There are three types of mammal hedgehogs, Sonic Hedgehog, Indian Hedgehog, and Desert hedgehog. The most studied among them is the Sonic Hedgehog (SHH) signaling pathway that can be expressed systemically.

The Sonic Hedgehog signaling pathway begins by the binding of Sonic Hedgehog to Patched1 (Ptch1), a 12-membrane-through receptor in the cell membrane, and Ptch1 inhibits SMO, a 7-membrane-type Gli protein-binding receptor. When Sonic Hedgehog binds to, Ptch1 loses its action to inhibit SMO. As a result, SMO is activated, Gli, which is a transcription factor, is activated by the activated SMO, and the expression of the hedgehog target gene is activated. That is, the hedgehog acting as a ligand binds to the Ptch1 receptor present in the cell membrane and promotes the migration of SMO, another GPCR-like membrane transmembrane protein whose activity is inhibited by Phch1, to the primary ciliary, resulting in hedgehog signaling. It promotes the activity of the Gli transcription factor, a factor.

The way in which SMO modulates Gli transcription factor activity is by controlling the Sufu, which inhibits the migration of Gli transcription factors into the nucleus at the lower signaling stage by the hedgehog, or the stability of the Gli protein in the primary cilia. Increase the expression of the inactive Gli to the state of transcriptional activity to induce an increase in the expression of the gene hedge-hog acted by the Gli transcription factor to proceed the cell migration, proliferation and differentiation process.

There are a wide variety of cancers with hedgehog signaling involved. Examples include basal cell carcinoma, medulloblastoma, glioblastoma, breast cancer, prostate cancer, malignant melanoma, lung cancer, Pancreatic cancer.

Clinical studies and hypothesis testing of the causal relationship between cancer development and hedgehog signaling abnormalities in humans are well established. Recently, as the groundbreaking results associated with the development of cancer, research has been actively conducted on the occurrence and recurrence of cancer by cancer stem cells. Appropriate control or removal of cancer stem cells has emerged as an important cancer treatment method, and since cancer stem cells share the characteristics of stem cells in developmental stages, hedgehog signaling acts as an important regulator.

Based on this fact, as a target for the treatment of cancer, a method that plays an important role in the growth and differentiation of cells in the developmental stage and uses well-defined developmental regulatory factors as a control target of cancer stem cells has emerged as a cancer treatment. Therefore, the development of new drugs that can control developmental control factors such as hedgehog is a new concept target, and Erivedge (Vismodegib), the first-generation drug, was released in 2012 and its efficacy has been clinically proven.

At the same time, hedgehog also acts as a factor regulating the function of stromal cells that regulate the microenvironment of cancer stem cells as well as cancer cells and cancer stem cells. Therefore, inhibiting hedgehog signaling is an attractive and effective as a number of target control drugs that can simultaneously control the growth of cancer cells at various stages as one drug.

The release of the first-generation drug, Vismodegib, validated the hypothesis that cancer could be controlled by hedgehog targets and produced significant ripple effects as clinical drug development targets. In clinical practice, Bismodib has also been studied in the treatment of various other cancers, including colorectal cancer, small cell lung cancer, gastric cancer, pancreatic cancer, medulloblastoma and chondrosarcoma. Recently, WO 2010/147917 discloses a hedgehog signaling pathway inhibitor for the treatment of various cancers, and Novartis Oncology has finalized its approval for the SMO receptor inhibitor Sonidegib as a treatment for basal cell carcinoma. Accordingly, inhibition of abnormal hedgehog signaling pathways and inhibition of SMO expression have emerged as desirable targets for anticancer therapy.

Inhibition of the hedgehog signaling pathway using small molecules has become an important target for clinicians to treat clinically significant cancers such as solid tumors through reversal or control of abnormal cell growth. However, as an effective treatment for various cancer types, effective hedgehog signaling pathway inhibitors and SMO inhibitors are still needed (see Korean Patent Publication No. 10-1217316).

The present invention has been made to solve the above problems in the prior art, the present inventors have made diligent efforts to develop an anticancer agent having an SMO inhibitory activity, as a result of confirming the excellent SMO inhibitory activity of the novel compounds according to the present invention The invention has been completed.

Accordingly, it is an object of the present invention to provide a novel compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

Another object of the present invention is to provide a novel compound represented by the following formula (2) or a pharmaceutically acceptable salt thereof.

[Formula 2]

In addition, another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer diseases comprising the novel compound of Formula 1 or Formula 2 or a pharmaceutically acceptable salt thereof as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problem, another task that is not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the above object, the present invention provides a novel compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Chemical Formula 1,

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이고;R1 is

, or

ego;

또는

이다.R2 is

or

to be.

The present invention also provides a novel compound represented by the following formula (2) or a pharmaceutically acceptable salt thereof.

[Formula 2]

In Chemical Formula 2,

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이고;R1 is Cl,

,

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or

ego;

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또는

이다.R2 is

,

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,

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or

to be.

As a specific example, the compound represented by Formula 1 may be selected from the group consisting of the following compounds.

2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -4- (4- (trifluoromethoxy) phenyl) isoindolin-1-one,

2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -4- (pyridin-3-yl) isoindolin-1-one,

4- (2,4-difluorophenyl) -2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) isoindolin-1-one,

2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyrimidin-4-yl) -4- (4- (trifluoromethoxy) phenyl) isoindolin-1-one.

As a specific example, the compound represented by Formula 2 may be selected from the group consisting of the following compounds.

8-chloro-3- (4-fluorophenyl) quinazolin-4 (3H) -one,

8-chloro-3- (4-morpholinophenyl) quinazolin-4 (3H) -one,

8-chloro-3- (6-morpholinopyridin-3-yl) quinazolin-4 (3H) -one,

8-chloro-3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) quinazolin-4 (3H) -one,

8-chloro-3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyrimidin-4-yl) quinazolin-4 (3H) -one,

3- (4-morpholinophenyl) -8-phenylquinazolin-4 (3H) -one,

8- (3-methoxyphenyl) -3- (4-morpholinophenyl) quinazolin-4 (3H) -one,

3- (4-morpholinophenyl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one,

3- (6-morpholinopyridin-3-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8-phenylquinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one ,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (naphthalen-1-yl) quinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8-p-tolylquinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4-methoxyphenyl) quinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4- (methylsulfonyl) phenyl) quinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4-fluorophenyl) quinazolin-4 (3H) -one,

3- (5-((2S, 6R) -2,6-dimethylmorpholino) pyridin-2-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one ,

3- (6- (4-methylpiperazin-1-yl) pyridin-3-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyrimidin-4-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H)- On,

3- (6- (4-methylpiperidin-1-yl) pyrimidin-4-yl) -8- (4 (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one,

3- (2-morpholinoethyl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one,

3- (3- (1H-imidazol-1-yl) propyl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one.

The present invention also provides a pharmaceutical composition for preventing or treating cancer diseases comprising the compound of Formula 1 or Formula 2 as an active ingredient.

The present invention also provides a health functional food composition for improving cancer disease comprising the compound of Formula 1 or Formula 2 as an active ingredient.

In one embodiment of the invention, the cancer is basal cell carcinoma, medulloblastoma, rhabdomyosarcoma, chondroma, melanoma, small cell lung cancer, non-small cell lung cancer, B-cell lymphoma, multiple myeloma, brain cancer, esophageal cancer, breast cancer, ovary Cancer, stomach cancer, colorectal cancer, liver cancer, kidney cancer, head and neck cancer, mesothelioma, soft tissue sarcoma, osteosarcoma, testicular cancer, prostate cancer, pancreatic cancer, bone cancer, bone metastasis, acute leukemia, chronic leukemia, glioma, Hodgkins disease, cutaneous melanoma , Bladder cancer, endocrine cancer, parathyroid cancer, thyroid cancer, cervical cancer, endometrial cancer, ovarian cancer, skin cancer, renal cell carcinoma, pituitary adenoma, spinal axis tumor, uterine cancer, gastric cancer and biliary tract cancer.

In another embodiment of the invention, the composition may inhibit the hedgehog signaling pathway (Hedgehog signaling pathway).

In another embodiment of the present invention, the composition may inhibit SMO (Smoothened) activity.

In another embodiment of the present invention, the composition may inhibit Gli1 expression.

In another embodiment of the present invention, the composition may inhibit the hedgehog signaling pathway for the SMO (smoothened) variant (D477H).

The present invention also provides a method for treating a cancer disease comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound of Formula 1, Formula 2, or a pharmaceutically acceptable salt thereof. to provide.

In addition, the present invention provides a therapeutic use of cancer diseases of the pharmaceutical composition comprising a compound of Formula 1, Formula 2 or a pharmaceutically acceptable salt thereof.

The present invention relates to a novel compound having an SMO inhibitory activity, and exhibits excellent anticancer effect through the regulation of the hedgehog signaling pathway and the inhibitory activity of SMO activity of isoindolinone derivatives and quinazolinone derivative compounds. Bar, there is an advantage that can prevent, ameliorate or treat cancer diseases.

1 shows the types of drugs that inhibit SMO currently being developed or released.

Figure 2 is a diagram showing the IC ₅₀ analyzed the degree of hedgehog signal inhibition of the four isoindolinone derivatives (Isoindolinone).

FIG. 3 is a diagram showing IC ₅₀ of Hedgehog signal inhibition level of 22 quinazolinone derivatives. FIG.

FIG. 4 is a diagram showing representative examples (Compound 14 and Compound 20) of quinazolinone derivatives that inhibit Hedgehog signaling activated by SHh (Sonic Hedgehog) ligand.

FIG. 5 is a diagram showing a representative example of a quinazolinone derivative that inhibits signal transduction in the cells from which Ptch1 has been removed without the SHh ligand, thereby increasing Gli1 protein expression.

Figure 6 shows that Quinazolinone derivatives, like Vismodegib and Sonidegib, inhibit the migration of SMO proteins, which are sub-action genes of hedgehog signaling activity, to the cilia.

7 is a result of quantitative analysis of the degree of cilia migration inhibition of SMO protein shown in FIG.

Figure 8 shows that Quinazolinone derivatives, like Vismodegib and Sonidegib, inhibit the process of Gli2, a downstream action transcription factor of hedgehog signaling activity, to the cilia.

Figure 9 is a comparison of the degree of inhibition of the hedgehog signaling pathway for SMO (D477H) variants that cause resistance to Vismodegib for Vismodegib, Sonidegib and Quinazolinone derivatives.

The present inventors, when treated with the novel compound prepared in Example, based on the fact that it can inhibit hedgehog signaling, SMO inhibition, Gli transcription factor inhibition and SMO variant (D477H) inhibitory effect of the compound The present invention was specifically confirmed, and based on this, the present invention was completed.

Hereinafter, the present invention will be described in detail.

The present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Chemical Formula 1,

,

또는

이고;R1 is

,

or

ego;

또는

이다.R2 is

or

to be.

Preferred examples of the compound represented by Formula 1 according to the present invention are as follows.

2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -4- (4- (trifluoromethoxy) phenyl) isoindolin-1-one,

2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -4- (pyridin-3-yl) isoindolin-1-one,

4- (2,4-difluorophenyl) -2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) isoindolin-1-one,

2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyrimidin-4-yl) -4- (4- (trifluoromethoxy) phenyl) isoindolin-1-one.

The present invention also provides a compound represented by the following formula (2) or a pharmaceutically acceptable salt thereof.

[Formula 2]

In Chemical Formula 2,

,

,

,

,

,

,

또는 이고;R1 is Cl,

,

,

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or ego;

,

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또는

이다.R2 is

,

,

,

,

,

,

,

,

or

to be.

Preferred examples of the compound represented by Formula 2 according to the present invention are as follows.

8-chloro-3- (4-fluorophenyl) quinazolin-4 (3H) -one,

8-chloro-3- (4-morpholinophenyl) quinazolin-4 (3H) -one,

8-chloro-3- (6-morpholinopyridin-3-yl) quinazolin-4 (3H) -one,

8-chloro-3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) quinazolin-4 (3H) -one,

8-chloro-3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyrimidin-4-yl) quinazolin-4 (3H) -one,

3- (4-morpholinophenyl) -8-phenylquinazolin-4 (3H) -one,

8- (3-methoxyphenyl) -3- (4-morpholinophenyl) quinazolin-4 (3H) -one,

3- (4-morpholinophenyl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one,

3- (6-morpholinopyridin-3-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8-phenylquinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one ,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (naphthalen-1-yl) quinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8-p-tolylquinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4-methoxyphenyl) quinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4- (methylsulfonyl) phenyl) quinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4-fluorophenyl) quinazolin-4 (3H) -one,

3- (5-((2S, 6R) -2,6-dimethylmorpholino) pyridin-2-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one ,

3- (6- (4-methylpiperazin-1-yl) pyridin-3-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one,

3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyrimidin-4-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H)- On,

3- (6- (4-methylpiperidin-1-yl) pyrimidin-4-yl) -8- (4 (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one,

3- (2-morpholinoethyl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one,

3- (3- (1H-imidazol-1-yl) propyl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one.

As used herein, the term "pharmaceutically acceptable" is suitable for use in contact with the tissue of a subject (eg, a human being) because the benefit / risk ratio is reasonable without excessive toxicity, irritation, allergic reactions or other problems or complications. A compound or composition is within the scope of sound medical judgment.

As used herein, the term "salt" is useful for acid addition salts formed with pharmaceutically acceptable free acids. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. Obtained from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids. Such pharmaceutically nontoxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, and iodide. Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suverate , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, meth Oxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesul Nate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1- Sulfonates, naphthalene-2-sulfonates or mandelate.

The acid addition salts according to the present invention can be dissolved in conventional methods, for example, by dissolving a compound represented by the formula (1) or (2) in an excess of an aqueous solution of an acid, which salt is a water miscible organic solvent such as methanol, ethanol, acetone or It can be prepared by precipitation with acetonitrile. It may also be prepared by evaporating the solvent or excess acid from the mixture and then drying or by suction filtration of the precipitated salt.

Bases can also be used to make pharmaceutically acceptable metal salts. Alkali metal or alkaline earth metal salts are obtained, for example, by dissolving the compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salt as the metal salt. The corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg silver nitrate).

In one embodiment of the present invention, as a result of confirming the SMO inhibitory activity of the compound of Formula 1 or Formula 2, the compound was found to inhibit the activity of most SMO, in particular, quinazoli represented by Compound 14 and Compound 20 In the case of the non-derivative compound, it was found that the SMO inhibitory activity was superior to that of the positive control Vismodegib (see Example 29).

Therefore, the compound of Formula 1 or Formula 2 according to the present invention inhibits SMO (Smoothened) activity through the regulation of the hedgehog signaling pathway (Hedgehog signaling pathway), and inhibits the expression of the transcription factor Gli1 bar, anticancer activity is required It can be used for a variety of purposes and uses.

Accordingly, the present invention provides a composition for the prevention, improvement or treatment of cancer diseases containing the compound of Formula 1, Formula 2 or a pharmaceutically acceptable salt thereof as an active ingredient, the composition is a pharmaceutical composition and health function It may include all food compositions.

As used herein, the term "prevention" means any action that inhibits or delays the onset of a cancer disease by administration of a pharmaceutical composition according to the invention.

As used herein, the term "treatment" means any action that improves or advantageously changes to a cancer disease by administration of the pharmaceutical composition according to the present invention.

In the present invention, the cancer disease is basal cell carcinoma, medulloblastoma, rhabdomyosarcoma, chondroma, melanoma, small cell lung cancer, non-small cell lung cancer, B-cell lymphoma, multiple myeloma, brain cancer, esophageal cancer, breast cancer, ovarian cancer, gastric cancer, Colorectal cancer, liver cancer, kidney cancer, head and neck cancer, mesothelioma, soft tissue sarcoma, osteosarcoma, testicular cancer, prostate cancer, pancreatic cancer, bone cancer, bone metastasis, acute leukemia, chronic leukemia, glioma, Hodgkins disease, cutaneous melanoma, bladder cancer, endocrine It may be at least one selected from cough cancer, parathyroid cancer, thyroid cancer, cervical cancer, endometrial cancer, ovarian cancer, skin cancer, renal cell carcinoma, pituitary adenoma, spinal axis tumor, uterine cancer, gastric cancer and biliary tract cancer, but is not limited thereto. no. In addition to the diseases exemplified herein, diseases associated with hedgehog signaling pathway regulation known in the art are all considered to be included in cancer diseases that can be prevented or treated with a compound having the structure of Formula 1 or 2 of the present invention. . In one specific example, the cancer may be any cancer caused by hedgehog signaling hyperactivity.

In one embodiment, when the anticancer composition according to the present invention is used in the form of a pharmaceutical composition, the pharmaceutical composition may contain a pharmaceutically effective amount of a compound of Formula 1 or Formula 2 or a pharmaceutically acceptable salt thereof. It may further comprise a pharmaceutically acceptable carrier, excipient or diluent comprising the composition containing as a component. The pharmaceutically effective amount herein refers to an amount sufficient to prevent or treat the symptoms of cancer disease.

The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient. At this time, the pharmaceutically acceptable carrier is commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose , Polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. In addition to the above components, it may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like.

The pharmaceutical compositions of the present invention can be administered orally or parenterally (eg, applied intravenously, subcutaneously, intraperitoneally or topically) according to the desired method, and the dosage is determined by the condition and weight of the patient, Depending on the extent, drug form, route of administration, and time, it may be appropriately selected by those skilled in the art.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. As used herein, the term “pharmaceutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level refers to the type of disease, the severity, the activity of the drug, It may be determined according to the sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of treatment, factors including the drug used concurrently and other factors well known in the medical field. The pharmaceutical compositions according to the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered as single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the pharmaceutical composition of the present invention may vary depending on the age, sex, condition, weight of the patient, the absorption of the active ingredient in the body, the inactivation rate and excretion rate, the type of disease, the drug used in general 0.001 to 150 mg, preferably 0.01 to 100 mg, per kg of body weight may be administered daily or every other day, or divided into 1 to 3 times a day. However, since the dose may be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, age, etc., the above dosage does not limit the scope of the present invention by any method.

In addition, the present invention provides a method for treating a cancer disease comprising administering the pharmaceutical composition to a subject. As used herein, "individual" means a subject in need of treatment for a disease, and more specifically, a mammal, such as a primate, mouse, dog, cat, horse and cow, which is human or non-human. .

In another embodiment, the anticancer composition according to the present invention may be used in the form of a dietary supplement composition utilized in the manufacture of foods, such as a main ingredient, a subsidiary ingredient, a food additive, a functional food or a beverage, which are effective in improving cancer diseases. have.

In the above, the food means a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through some processing process. It includes all additives, functional foods and drinks.

Examples of the food to which the composition according to the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and functional foods. In addition, the food includes special nutritional products (e.g., formulated milk, young, infant food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g., ramen, noodles, etc.), breads, health supplements, seasoned foods ( For example, soy sauce, miso, red pepper paste, mixed soy sauce), sauces, sweets (e.g. snacks), candy, chocolates, gums, ice creams, dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (Various kimchi, pickles, etc.), beverages (e.g., fruit drinks, vegetable drinks, soy milk, fermented beverages, etc.), natural seasonings (e.g., ramen soup, etc.), but are not limited thereto. The additive may be prepared by a conventional manufacturing method.

In addition, the functional food is a biological defense rhythm control, disease prevention and recovery of a food group or a food composition that has added value to the food by using physical, biochemical, biotechnological techniques, etc. to function and express the function of the food for a specific purpose. It means a food that is designed and processed to fully express the body control function on the living body, preferably, it may be a health functional food. The functional food may include food acceptable food additives, and may further include appropriate carriers, excipients and diluents commonly used in the manufacture of functional foods.

In addition, in the present invention, the beverage refers to a generic term of drinking to quench thirst or enjoy a taste and includes a functional beverage. The beverage includes, as an essential component, a composition containing a compound of Formula 1 or Formula 2 or a pharmaceutically acceptable salt thereof for the prevention or amelioration of cancer symptoms. Various flavors or natural carbohydrates and the like may be included as additional ingredients.

In addition, foods containing the composition of the present invention in addition to those described above include flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and fillers (such as cheese, chocolate), Pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like, which components may be independently or It can be used in combination.

In the food containing the composition for food according to the present invention, the amount of the composition may comprise 0.001% to 90% by weight of the total food weight, preferably 0.1% to 40% by weight In the case of a beverage, it may be included in a ratio of 0.001g to 2g, preferably 0.01g to 0.1g based on 100ml, and may be less than the above range for long-term intake, but the active ingredient has no problem in terms of safety. Since it may be used in an amount greater than the above range because it is not limited to the above range.

Hereinafter, preferred examples are provided to aid in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the following examples.

[ Example ]

Example 1 (Compound 1) 2- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-3-yl) -4- (4- (trifluoromethoxy) phenyl) iso Preparation of Indolin-1-one

Step 1: ethyl 3- bromo- 2- methyl Preparation of benzoate

An azeotropic mixture was prepared by refluxing a mixture of 3-bromo-2-methyl benzoic acid (2.00 g, 9.30 mmol), concentrated sulfuric acid (0.20 mL) and ethanol (3.00 ml) under benzene (10.0 mL) for 18 hours. The reaction mixture was cooled to room temperature and washed with saturated sodium bicarbonate solution and water. The organic phase was recovered, dried and concentrated. The crude product was purified by flash column chromatography (Hex: EA 5: 1) to give a colorless liquid as 1.99 g (88.0%) of pure compound.

Step 2: Preparation of ethyl 3-bromo-2- (bromomethyl) benzoate

To the mixture of ethyl 3-bromo-2-methyl benzoate (1.99 g, 8.18 mmol) (step 1) under benzene was added NBS (1.75 g, 9.82 mmol) and AIBN (0.14 g, 0.81 mmol). After refluxing the reaction solution for 5 hours, the solution was cooled to room temperature and the precipitate was filtered off. The filtrate was saturated with Na ₂ S ₂ O ₃ Washed with solution and water. The organic phase was dried and concentrated, then purified by flash column chromatography (Hex: EA 5: 1) to give a colorless oil as 2.54 g (96.0%) of pure compound.

Step 3: Preparation of 4 - bromo- 2- (6-(( 2S, 6R ) -2,6 -dimethylmorpholino ) pyridin-3-yl) isoin dolin-1-one

Ethyl 3-bromo-2- (bromomethyl) benzoic acid (0.50 g, 1.55 mmol) mixture (step 2), 6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-amine (0.39 g, 1.89 mmol) and DIPEA (0.18 ml, 1.89 mmol) were sealed with ethanol in a closed tube. The reaction mixture was heated to 100 ° C. overnight, then cooled to room temperature and ethanol was removed under reduced pressure. The residue was dissolved in ethyl acetate and washed with saturated sodium bicarbonate solution. The combined organic phases were dried, filtered and concentrated. The crude product was purified by flash column chromatography (MC: EA: Hex 1: 1: 2) to afford a white cream solid with 0.48 g (77.0%) of pure compound.

Step 4: 2- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-3-yl) -4- (4- ( trifluoromethoxy ) phenyl) isoindolin-1- Manufacture of

4-bromo-2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) isoindolin-1-one mixture (step 3), (4- ( Trifluoromethoxy) phenyl) methanediol (1.5 equiv) and 2 M aqueous sodium bicarbonate solution were sealed with a DME in a closed tube. The solution was bubbled under argon for 15 minutes and palladium catalyst (15.0 mg, 5 mol%) was added. The reaction solution was further bubbled under argon for 5 minutes, the sealed tube was heated to 130 ° C. overnight, then cooled to room temperature and the solvent was removed under reduced pressure. The residue was dissolved in ethyl acetate and washed with water. The combined organic phases were dried, filtered and concentrated. The crude product was purified by flash column chromatography to give pure compound.

Cream white solid (93%);

mp 191.4-192.6 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.44 (d, J = 2.4 Hz, 1H), 8.10 (dd, J = 3.0, 9.4 Hz, 1H), 7.91 (dd, J = 2.2, 6.6 Hz, 1H) , 7.61-7.56 (m, 2H), 7.53-7.51 (m, 2H), 7.36 (d, J = 8.4 Hz, 2H), 6.67 (d, J = 9.2 Hz, 1H), 4.81 (s, 2H), 4.03-3.99 (m, 2H), 3.74-3.69 (m, 2H), 2.54-2.48 (m, 2H), 1.27 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 167.08, 156.55, 149.06, 139.73, 137.81, 137.14, 135.77, 133.51, 132.06, 130.68, 129.53, 129.22, 127.00, 123.36, 121.46, 106.89, 71.52, 51.00 18.98;

HRMS (EI) m / z calcd for C ₂₆ H ₂₅ F ₃ N ₃ O ₃ [M ⁺ ] 484.1848, found: 484.1883.

Example 2: (Compound 2) 2- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-3-yl) -4- (pyridin-3-yl) isoindolin-1 Manufacture of

Pure compound was obtained in the same manner as in Example 1, except that pyridin-3-ylmethanediol was used instead of (4- (trifluoromethoxy) phenyl) methanediol of step 4.

Cream white solid (92%);

mp 219.9-203.0 ° C;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.79 (d, J = 1.6 Hz, 1H), 8.69 (dd, J = 1.6, 4.8 Hz, 1H), 8.45 (d, J = 2.4 Hz, 1H), 8.08 (dd, J = 4.8, 7.2 Hz, 1H), 7.96 (dd, J = 1.6, 7.2 Hz, 1H), 7.84-7.81 (m, 1H), 7.66-7.51 (m, 2H), 7.47-7.44 (m , 1H), 6.69 (d, J = 9.2 Hz, 1H), 4.85 (s, 2H), 4.04-4.00 (m, 2H), 3.75-3.70 (m, 2H), 2.55-2.49 (m, 2H), 1.27 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 166.95, 156.60, 149.41, 148.86, 139.84, 138.10, 135.40, 134.25, 133.69, 133.57, 132.17, 130.76, 129.42, 126.88, 123.83, 123.80, 106.91, 71.53, 71.53, 71. 50.61, 18.99;

HRMS (EI) m / z calcd for C ₂₄ H ₂₅ N ₄ O ₂ [M ⁺ ] 401.1978, found: 401.2008.

Example 3: (Compound 3) 4- (2,4 -difluorophenyl ) -2- (6-(( 2S, 6R ) -2,6- dimethyl morpholino) pyridin-3-yl) isoin Preparation of Dolin-1-one

A pure compound was prepared in the same manner as in Example 1, except that (2,4-difluorophenyl) methanediol was used instead of (4- (trifluoromethoxy) phenyl) methanediol of step 4. Got it.

Cream white solid (75%);

mp 176.0177.2 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.42 (d, J = 2.8 Hz, 1H), 8.12 (dd, J = 3.0, 9.0 Hz, 1H), 7.95 (dd, J = 1.0, 7.4 Hz, 1H) , 7.60 (d, J = 7.6 Hz, 1H), 7.53 (d, J = 6.8 Hz, 1H), 7.407.34 (m, 1H), 7.056.96 (m, 2H), 6.68 (d, J = 9.6 Hz, 1H), 4.71 (s, 2H), 4.034.00 (m, 2H), 3.753.70 (m, 2H), 2.552.49 (m, 2H), 1.27 (d, J = 6.4 Hz, 6H) .

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 167.07, 156.55, 139.72, 139.25, 133.27, 133.18, 131.95, 131.90, 131.85, 131.80, 130.73, 130.48, 128.91, 127.10, 123.79, 112.20, 112.16, 111.95. 106.92, 104.85, 104.58, 104.33, 71.53, 51.03, 50.54, 50.47, 18.99;

HRMS (EI) m / z calcd for C ₂₅ H ₂₄ F ₂ N ₃ O ₂ [M ⁺ ] 436.1837, found: 436.1849.

Example 4: (Compound 4) 2- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyrimidin-4-yl) -4- (4- (trifluoromethoxy) phenyl) Preparation of Isoindolin-1-one

Step 1: ethyl 3- bromo- 2- methyl Preparation of benzoate

Prepared in the same manner as in Example 1 (Step 1), to obtain a pure compound.

¹ H-NMR (400Mz, CDCl ₃ ) δ 7.727.66 (s, 2H), 7.09 (t, J = 7.8 Hz, 1H), 4.394.33 (m, 2H), 2.62 (s, 3H), 1.39 ( t, J = 7.0 Hz, 3H).

Step 2: Ethyl 2- Methyl- 4 '-( trifluoromethoxy ) biphenyl- 3- carboxylate Produce

Prepared in the same manner as in Example 1 (Step 4), to obtain a pure compound.

¹ H-NMR (400 Mz, CDCl ₃ ) δ 7.81 (dd, J = 1.8, 7.4 Hz, 1H), 7.33-7.25 (m, 6H), 4.39 (q, J = 7.2 Hz, 2H), 2.39 (s, 3H), 1.31 (t, J = 7.0 Hz, 3H).

Step 3: Preparation of ethyl 2- ( bromomethyl ) -4 '-( trifluoromethoxy ) biphenyl- 3- carboxylate

Prepared in the same manner as in Example 1 (Step 2), to obtain a pure compound.

¹ H-NMR (400Mz, CDCl ₃ ) δ 7.93 (dd, J = 1.8, 8.2 Hz, 1H), 7.47-7.29 (m, 6H), 4.84 (s, 2H), 4.44 (q, J = 7.2 Hz, 2H), 1.44 (t, J = 7.2 Hz, 3H).

Step 4: 2- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyrimidin-4-yl) -4- (4- ( trifluoromethoxy ) phenyl) isoindolin- Preparation of 1-On

Ethyl 2- (bromomethyl) -4 '-(trifluoromethoxy) biphenyl-3-carboxylate (0.10 g, 0.28 mmol) mixture (step 3), 6-((2S, 6R) -2,6 Dimethylmorpholino) pyrimidin-4-amine (0.07 g, 0.34 mmol) and DIPEA (0.06 ml, 0.34 mmol) were sealed with ethanol in a closed tube. The reaction mixture was heated to 100 ° C. overnight, then cooled to room temperature and ethanol was removed under reduced pressure. The residue was dissolved in ethyl acetate and washed with saturated sodium bicarbonate solution. The combined organic phases were dried, filtered and concentrated. The crude product was purified by flash column chromatography (MC: EA: Hex 1: 1: 2) to afford a white cream solid with 0.02 g (14.7%) of pure compound.

White solid (15%);

mp 249.0-250.1 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.43 (s, 1H), 7.94-7.92 (m, 2H), 7.64-7.60 (m, 2H), 7.55-7.51 (m, 2H), 7.36 (d, J = 8.0 Hz, 2H), 5.10 (s, 2H), 4.24 (brs, 2H), 3.71-3.63 (m, 2H), 2.67-2.61 (m, 2H), 1.28 (d, J = 6.4 Hz, 6H) .

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 168.48, 162.91, 157.43, 157.14, 138.66, 136.90, 136.41, 133.24, 133.21, 129.62, 129.24, 123.54, 121.55, 90.07, 76.69, 71.54, 49.48, 49.40, 49.40 ;

HRMS (EI) m / z calcd for C ₂₅ H ₂₄ F ₃ N ₄ O ₃ [M ⁺ ] 485.1801, found: 485.1837.

Example 5: Preparation of (Compound 5) 8 -Chloro- 3- (4- fluorophenyl ) quinazolin- 4 (3H) -one

3-Chloroanthranilic acid was sealed in a closed tube with orthoformate triethyl (1.2 equiv), 4-fluoroaniline (1.2 equiv) and THF, and the reaction mixture was heated to 110 ° C. for 18 hours. It was cooled to room temperature, saturated sodium bicarbonate solution was added, and the crude product was extracted with ethyl acetate. The combined organic phases were dried with magnesium sulfate, filtered and concentrated under reduced pressure, which was purified by flash column chromatography.

White solid (22%);

mp 232.5233.2 ° C;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.29 (d, J = 8.4 Hz, 1H), 8.20 (s, 1H), 7.90 (d, J = 7.6 Hz, 1H), 7.49 (t, J = 7.8 Hz , 1H), 7.427.39 (m, 2H), 7.287.23 (m, 2H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 164.02, 161.54, 160.22, 146.51, 144.56, 135.04, 133.02, 132.98, 132.06, 128.87, 128.78, 127.94, 126.06, 123.89, 116.99, 116.75;

HRMS (EI) m / z calcd for C ₁₄ H ₉ ClFN ₂ O [M ⁺ ] 275.0387, found: 275.0382.

Example 6: Preparation of (Compound 6) 8 -Chloro- 3- (4 -morpholinophenyl ) quinazolin- 4 (3H) -one

A pure compound was obtained in the same manner as in Example 5, except that 4-morpholinoaniline was used instead of 4-fluoroaniline.

White solid (33%);

mp 275.0275.8 ° C;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.28 (dd, J = 1.4, 8.2 Hz, 1H), 8.22 (s, 1H), 7.88 (dd, J = 1.4, 7.8 Hz, 1H), 7.46 (t, J = 7.8 Hz, 1H), 7.30 (d, J = 12.0 Hz, 2H), 7.03 (d, J = 12.0 Hz, 2H), 3.89 (t, J = 4.8 Hz, 3H), 3.25 (t, J = 5.0 Hz, 3H);

HRMS (EI) m / z calcd for C ₁₈ H ₁₇ ClN ₃ O ₂ [M ⁺ ] 342.1009, found: 342.0997.

Example 7: (Compound 7) 8- chloro-3- (6-morpholinophenyl-3-yl) quinazoline - 4 (3H) - Preparation of a whole

A pure compound was obtained in the same manner as in Example 5, except that 6-morpholinopyridin-3-amine was used instead of 4-fluoroaniline.

White solid (14%);

mp 264.9266.2 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.28 (d, J = 8.0 Hz, 1H), 8.19 (s, 2H), 7.89 (d, J = 7.2 Hz, 1H), 7.59 (dd, J = 2.8, 9.2 Hz, 1H), 7.48 (t, J = 7.8 Hz, 1H), 6.76 (d, J = 9.2 Hz, 2H), 3.84 (t, J = 4.8 Hz, 4H), 3.61 (t, J = 5.0 Hz , 4H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 160.51, 159.14, 146.75, 145.37, 144.66, 136.17, 134.94, 132.01, 127.82, 126.04, 124.19, 123.85, 106.59, 66.63, 45.33);

HRMS (EI) m / z calcd for C ₁₇ H ₁₆ ClN ₄ O ₂ [M ⁺ ] 343.0962, found: 343.0984.

Example 8 Preparation of (Compound 8) 8 -Chloro- 3- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin- 3 - yl) quinazolin-4 (3H) -one

A pure compound was obtained in the same manner as in Example 5, except that 6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-amine was used instead of 4-fluoroaniline.

White solid (30%);

mp 200.0200.8 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.26 (dd, J = 1.4, 7.8 Hz, 1H), 8.18 (s, 1H), 8.17 (d, J = 2.8 Hz, 1H), 7.88 (dd, J = 1.6, 8.0 Hz, 1H), 7.58 (dd, J = 2.6, 9.0 Hz, 1H), 7.46 (t, J = 7.8 Hz, 1H), 6.75 (d, J = 9.2 Hz, 1H), 4.154.11 ( m, 2H), 3.743.70 (m, 2H), 2.652.59 (m, 2H), 1.29 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 160.47, 158.71, 146.76, 145.29, 144.61, 136.13, 134.89, 131.96, 127.76, 125.99, 123.86, 123.80, 106.61, 71.52, 50.45, 18.94;

HRMS (EI) m / z calcd for C ₁₉ H ₂₀ ClN ₄ O ₂ [M ⁺ ] 371.1275, found: 371.1284.

Example 9 (Compound 9) 8 -Chloro- 3- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyrimidin -4-yl) quinazolin-4 (3H) -one Manufacture

A pure compound was obtained in the same manner as in Example 5, except that 6-((2S, 6R) -2,6-dimethylmorpholino) pyrimidin-4-amine was used instead of 4-fluoroaniline. .

White solid (5%);

mp 212.8214.1 ° C;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.91 (s, 1H), 8.61 (s, 1H), 8.28 (dd, J = 1.4, 8.2 Hz, 1H), 7.89 (dd, J = 1.2, 7.6 Hz, 1H), 7.48 (t, J = 7.8 Hz, 1H), 7.25 (s, 1H), 4.254.11 (m, 2H), 3.723.64 (m, 2H), 2.69 (t, J = 11.6 Hz, 2H ), 1.28 (d, J = 6.0 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 162.61, 159.96, 158.36, 155.53, 144.64, 143.98, 135.33, 132.24, 127.89, 126.04, 123.63, 97.99, 71.46, 49.45, 18.81;

HRMS (EI) m / z calcd for C ₁₈ H ₁₉ ClN ₅ O ₂ [M ⁺ ] 372.1227, found: 372.1247.

Example 10 Preparation of (Compound 10) 3- (4 -morpholinophenyl ) -8 -phenylquinazolin- 4 (3H) -one

(1) step 1

Under DME and water, sodium carbonate (2 equiv) and phenylmethanediol (1.1 equiv) were added to the 7-bromoissatin solution. The reaction mixture was degassed with argon for 30 minutes, then Pd (PPh 3) 4 (5 mol%) was added and the solution was refluxed for 3 hours. After cooling the reaction solution to room temperature, the organic solvent was removed under reduced pressure. The aqueous phase was extracted with methylene chloride and the combined organic layers were dried with magnesium sulfate, filtered and concentrated. The crude product was purified by flash column chromatography (MC: methanol 50: 1).

Preparation of on-3 - (4-morpholinophenyl) -8-phenyl-quinazolin -4 (3H): (2) Step 2

Arylisatin (1 mmol) was dissolved in 5% NaOH (5 mL) and hydrogen peroxide (30% aqueous solution) (5 mL) was added drop-wise with stirring at room temperature. After stirring at 50 ° C. for 30 minutes, the solution was cooled to room temperature and acidified to pH 2 with 2N HCl solution. The white precipitate obtained was recovered and used directly in the next step. Trimethyl ortho formic acid (1.2 mmol), 4-morpholinoaniline (1.2 mmol) and THF were added together to the prepared acid and sealed in a sealed tube, and the resulting reaction mixture was heated to 110 ° C. for 18 hours. To this was added saturated sodium bicarbonate solution and after cooling to room temperature, the crude product was extracted with ethyl acetate. The combined organic phases were concentrated filtered under reduced pressure, dried over magnesium sulfate and purified by flash column chromatography (MC: methanol 40: 1).

White solid (12%);

mp 143.9144.5 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.40 (dd, J = 0.4, 9.6 Hz, 1H), 8.11 (s, 1H), 7.81 (dd, J = 1.6, 7.2 Hz, 1H), 7.627.57 ( m, 3H), 7.49 (t, J = 7.6 Hz, 2H), 7.437.25 (m, 3H), 7.02 (d, J = 8.8 Hz, 2H), 3.88 (t, J = 5.0 Hz, 4H), 3.23 (t, J = 4.6 Hz, 4H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.26, 151.47, 145.87, 145.18, 139.73, 138.38, 135.40, 130.29, 129.04, 128.09, 127.65, 127.59, 127.23, 126.62, 122.97, 115.83, 66.75, 48.76, 48.76

HRMS (EI) m / z calcd for C ₂₄ H ₂₂ N ₃ O ₂ [M ⁺ ] 384.1712, found: 384.1681.

Example 11: (Compound 11) 8- ( 3- methoxy-phenyl) -3- (4-morpholinophenyl) quinazolin - 4 (3H) - Preparation of a whole

A pure compound was obtained by the same method as Example 10, except that (3-methoxyphenyl) methanediol was used instead of phenylmethanediol of step 1.

White solid (12%);

mp 184.2185.5 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.40 (dd, J = 1.4, 7.8 Hz, 1H), 8.11 (s, 1H), 7.80 (dd, J = 1.6, 7.2 Hz, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.40 (t, J = 8.0 Hz, 1H), 7.327.28 (m, 2H), 7.197.16 (m, 2H), 7.02 (d, J = 8.8 Hz, 2H), 6.96 (dd, J = 2.4, 8.4 Hz, 1H), 3.88 (t, J = 5.8 Hz, 4H), 3.86 (s, 3H), 3.24 (t, J = 4.8 Hz, 4H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.25, 159.22, 151.49, 145.91, 145.20, 139.75, 139.60, 135.35, 129.07, 129.04, 127.59, 127.18, 126.70, 122.96, 122.75, 116.16, 115.85. 55.30, 48.77;

HRMS (EI) m / z calcd for C ₂₅ H ₂₄ N ₃ O ₃ [M ⁺ ] 414.1818, found: 414.1789.

Example 12: (Compound 12) 3- (4-morpholinophenyl) -8- (4- (trifluoromethoxy) Fe carbonyl) quinazolin -4 (3H) - Preparation of a whole

A pure compound was obtained by the same method as Example 10, except that (4- (trifluoromethoxy) phenyl) methanediol was used instead of phenylmethanediol of step 1.

White solid (7%);

mp 164.8166.0 ° C .;

¹ H-NMR (400 Mz, CDCl ₃ ) δ 8.43 (d, J = 8.4 Hz, 1H), 8.12 (s, 1H), 7.79 (d, J = 7.6 Hz, 1H), 7.66 (d, J = 8.4 Hz , 2H), 7.60 (t, J = 7.6 Hz, 1H), 7.32 (t, J = 9.2 Hz, 4H), 7.02 (d, J = 8.4 Hz, 2H), 3.89 (t, J = 4.8 Hz, 4H ), 3.24 (t, J = 4.8 Hz, 4H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.13, 151.55, 148.81, 146.10, 145.09, 138.18, 136.94, 135.32, 131.73, 128.90, 127.56, 127.31, 127.11, 123.10, 120.52, 119.25, 115.85, 66.75, 66.75

HRMS (EI) m / z calcd for C ₂₅ H ₂₁ F ₃ N ₃ O ₃ [M ⁺ ] 468.1535, found: 468.1510.

Example 13: Preparation of (Compound 13) 3- (6- morpholinopyridin- 3 - yl) -8- (4- ( trifluoromethoxy ) phenyl) quinazolin-4 (3H) -one

Except for using (4- (trifluoromethoxy) phenyl) methanediol instead of phenylmethanediol in step 1 and 6-morpholinopyridin-3-amine instead of 4-morpholinoaniline in step 2 And it was prepared in the same manner as in Example 10 to obtain a pure compound.

White solid (19 mg, 12%);

mp 135.7137.1 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.42 (dd, J = 1.6, 8.0 Hz, 1H), 8.19 (d, J = 2.8 Hz, 1H), 8.08 (s, 1H), 7.81 (dd, J = 1.6, 7.2 Hz, 1H), 7.767.59 (m, 4H), 7.34 (dd, J = 1.0, 8.6 Hz, 2H), 6.76 (d, J = 8.8 Hz, 1H), 3.84 (t, J = 5.0 Hz, 4H), 3.60 (t, J = 4.8 Hz, 4H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.13, 159.06, 148.82, 145.61, 145.31, 145.08, 138.32, 136.81, 136.23, 135.52, 131.72, 127.49, 127.08, 124.50, 122.90, 120.54, 106.60, 66.

HRMS (EI) m / z calcd for C ₂₄ H ₂₀ F ₃ N ₄ O ₃ [M ⁺ ] 469.1488, found: 468. 469.1497.

Example 14 Preparation of (Compound 14) 3- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-3-yl) -8-phenylquinazolin-4 (3H) -one

Prepared in the same manner as in Example 10, except that 6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-amine was used instead of 4-morpholinoaniline of Step 2 The compound was obtained.

White solid (21%);

mp 95.496.1 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.39 (dd, J = 1.4, 7.8 Hz, 1H), 8.17 (d, J = 2.4 Hz, 2H), 8.07 (s, 1H), 7.82 (dd, J = 1.4, 7.4 Hz, 1H), 7.617.58 (m, 4H), 7.49 (t, J = 7.4 Hz, 2H), 7.42 (t, J = 7.6 Hz, 1H), 6.74 (d, J = 8.8 Hz, 1H), 4.144.09 (m, 2H), 3.763.68 (m, 2H), 2.642.58 (m, 2H), 1.29 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.26, 158.62, 145.40, 145.25, 145.16, 139.84, 138.24, 136.25, 135.56, 130.25, 128.10, 127.70, 127.39, 126.56, 124.32, 122.76, 106.63, 71.53. 18.95;

HRMS (EI) m / z calcd for C ₂₅ H ₂₅ N ₄ O ₂ [M ⁺ ] 413.1978, found: 413.1950.

Example 15: (Compound 15) 3- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-3-yl ) -8- (4- (trifluoromethoxy) phenyl) quina Preparation of Zolin-4 (3H) -one

(4- (trifluoromethoxy) phenyl) methanediol is used instead of phenylmethanediol in step 1, and 6-((2S, 6R) -2,6-dimethyl instead of 4-morpholinoaniline in step 2 Except that morpholino) pyridin-3-amine was used, the compound was prepared in the same manner as in Example 10, to obtain a pure compound.

White solid (14%);

mp 188.4189.6 ° C .;

¹ H-NMR (400 Mz, CDCl ₃ ) δ 8.42 (dd, J = 1.6, 8.4 Hz, 1H), 8.17 (d, J = 2.0 Hz, 2H), 8.08 (s, 1H), 7.80 (dd, J = 1.6, 7.2 Hz, 1H), 7.667.58 (m, 4H), 7.32 (d, J = 8.0 Hz, 2H), 6.75 (d, J = 9.2 Hz, 1H), 4.144.11 (m, 2H), 3.753.70 (m, 2H), 2.652.59 (m, 2H), 1.29 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.15, 158.69, 148.83, 145.65, 145.27, 145.10, 138.32, 136.83, 136.23, 135.51, 131.73, 127.49, 127.08, 124.20, 122.92, 120.54, 106.66. 18.97;

HRMS (EI) m / z calcd for C ₂₆ H ₂₄ F ₃ N ₄ O ₃ [M ⁺ ] 497.1801, found: 497.1776.

Example 16: (Compound 16) 3- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-3 -yl) -8- (naphthalen-1-yl) quinazolin-4 ( Preparation of 3H) -one

Naphthalen-1-ylboronic acid was used in place of phenylmethanediol in step 1, and 6-((2S, 6R) -2,6-dimethylmorpholino) pyridine-3-in place of 4-morpholinoaniline in step 2 Except for using the amine was prepared in the same manner as in Example 10 to obtain a pure compound.

White solid (16%);

mp 122.5123.7 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.48 (dd, J = 1.4, 8.2 Hz, 1H), 8.16 (d, J = 2.2 Hz, 1), 7.928.06 (m, 1H), 7.82 (dd, J = 1.2, 7.2 Hz, 1H), 7.687.58 (m, 3H), 7.527.43 (m, 3H), 7.397.35 (m, 1H), 6.72 (d, J = 8.2 Hz, 1H), 4.114 .09 (m, 2H), 3.723.69 (m, 2H), 2.632.56 (m, 2H), 1.28 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.29, 158.61, 146.51, 145.70, 145.26, 138.87, 136.73, 136.57, 136.25, 133.49, 132.37, 128.38, 128.36, 127.68, 127.23, 127.01, 126.19, 126.19, 126.19 125.23, 124.33, 122.53, 106.62, 71.54, 50.49, 18.95;

HRMS (EI) m / z calcd for C ₂₉ H ₂₇ N ₄ O ₂ [M ⁺ ] 463.2134, found: 463.2112.

Example 17: (Compound 17) 3- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-3-yl) -8-p-tolylquinazolin-4 (3H) -one Manufacture

4-tolylmethanediol is used instead of phenylmethanediol of step 1, and 6-((2S, 6R) -2,6-dimethylmorpholino) pyridine-3-in place of 4-morpholinoaniline of step 2 Except for using the amine was prepared in the same manner as in Example 10 to obtain a pure compound.

White solid (14%);

mp 158.7160.1 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.37 (dd, J = 1.4, 8.2 Hz, 1H), 8.17 (d, J = 2.8 Hz, 1H), 8.06 (s, 1H), 7.80 (dd, J = 1.4, 7.0 Hz, 1H), 7.617.56 (m, 2H), 7.517.49 (m, 2H), 7.30 (d, J = 8.0 Hz, 2H), 6.74 (d, J = 9.2 Hz, 1H), 4.144.10 (m, 2H), 3.763.69 (m, 2H), 2.642.58 (m, 2H), 1.29 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.33, 158.63, 145.31, 145.25, 139.91, 137.50, 136.28, 135.44, 135.32, 130.12, 128.88, 127.38, 126.33, 124.38, 122.75, 106.64, 71.55, 50.51, 50.51 22.69, 18.96;

HRMS (EI) m / z calcd for C ₂₆ H ₂₇ N ₄ O ₂ [M ⁺ ] 427.2134, found: 427.2114.

Example 18: (Compound 18) 3- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-3-yl) -8- (4-methoxyphenyl) quinazolin-4 ( Preparation of 3H) -one

(3-methoxyphenyl) methanediol is used instead of phenylmethanediol of step 1, and 6-((2S, 6R) -2,6-dimethylmorpholino) instead of 4-morpholinoaniline of step 2 A pure compound was obtained in the same manner as in Example 10 except that pyridin-3-amine was used.

White solid (18%);

mp 188.3189.5 ° C;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.36 (dd, J = 1.4, 7.8 Hz, 1H), 8.17 (d, J = 2.4 Hz, 1H), 8.07 (s, 1H), 7.80 (dd, J = 1.6, 7.2 Hz, 1H), 7.617.54 (m, 4H), 7.067.02 (m, 2H), 6.74 (d, J = 9.2 Hz, 1H), 4.144.11 (m, 2H), 3.87 (s , 3H), 3.753.70 (m, 2H), 2.642.58 (m, 2H), 1.29 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.33, 159.26, 158.63, 145.25, 145.24, 145.18, 139.50, 136.28, 135.35, 131.43, 130.56, 127.40, 126.13, 124.38, 122.80, 113.66, 106.64, 71. 50.51, 29.69, 18.96;

HRMS (EI) m / z calcd for C ₂₆ H ₂₇ N ₄ O ₃ [M ⁺ ] 443.2083, found: 443.2061.

Example 19: (Compound 19) 3- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-3-yl ) -8- (4- (methylsulfonyl) phenyl) quinazoline Preparation of -4 (3H) -one

(4- (methylsulfonyl) phenyl) methanediol is used instead of phenylmethanediol in step 1, and 6-((2S, 6R) -2,6-dimethylmol instead of 4-morpholinoaniline in step 2 A pure compound was obtained in the same manner as in Example 10, except that polyno) pyridin-3-amine was used.

White solid (15%);

mp 113.5115.0 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.47 (dd, J = 1.6, 8.0 Hz, 1H), 8.17 (d, J = 2.4 Hz, 1H), 8.098.05 (m, 3H), 7.857.81 ( m, 3H), 7.667.58 (m, 2H), 6.74 (d, J = 8.8 Hz, 1H), 4.144.11 (m, 2H), 3.753.71 (m, 2H), 3.19 (s, 3H) , 2.652.59 (m, 2H), 1.29 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 160.96, 158.71, 146.02, 145.25, 145.05, 143.95, 139.47, 137.63, 136.18, 135.55, 131.29, 127.90, 127.59, 127.21, 124.04, 123.02, 110.64, 54, 6666 50.48, 44.61, 29.70, 18.96;

HRMS (EI) m / z calcd for C ₂₆ H ₂₇ N ₄ O ₄ S [M ⁺ ] 491.1753, found: 491.1731.

Example 20: (Compound 20) 3- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-3-yl) -8- (4-fluorophenyl) quinazolin-4 ( Preparation of 3H) -one

(4-fluorophenyl) methanediol is used instead of phenylmethanediol of step 1, and 6-((2S, 6R) -2,6-dimethylmorpholino) pyridine instead of 4-morpholinoaniline of step 2 A pure compound was obtained in the same manner as in Example 10, except that 3-amine was used.

White solid (17%);

mp 138.9140.0 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.40 (dd, J = 1.6, 8.0 Hz, 1H), 8.17 (d, J = 2.0 Hz, 1H), 8.07 (s, 1H), 7.79 (dd, J = 1.6, 7.6 Hz, 1H), 7.617.56 (m, 4H), 7.207.15 (m, 2H), 6.74 (d, J = 9.2 Hz, 1H), 4.144.11 (m, 2H), 3.763.68 (m, 2H), 2.642.58 (m, 2H), 1.29 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 163.74, 161.29, 161.22, 158.66, 145.51, 145.27, 145.14, 138.77, 136.25, 135.46, 134.16, 134.13, 131.98, 131.90, 127.44, 126.74, 124.25, 122. 115.01, 106.65, 71.56, 50.50, 29.70, 18.96;

HRMS (EI) m / z calcd for C ₂₅ H ₂₄ FN ₄ O ₂ [M ⁺ ] 431.1883, found: 431.1874.

Example 21: (Compound 21) 3- (5-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-2-yl ) -8- (4- (trifluoromethoxy) phenyl) quina Preparation of Zolin-4 (3H) -one

(4- (trifluoromethoxy) phenyl) methanediol instead of phenylmethanediol in step 1, and 5-((2S, 6R) -2,6-dimethyl instead of 4-morpholinoaniline in step 2 Except for using morpholino) pyridin-2-amine was prepared in the same manner as in Example 10 to obtain a pure compound.

White solid (6%);

mp 146.5-147.6 ° C .;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.53 (s, 1H), 8.43 (dd, J = 1.4, 8.2 Hz, 1H), 8.20 (d, J = 2.8 Hz, 1H), 7.79 (dd, J = 1.6, 7.6 Hz, 1H), 7.73-7.65 (m, 3H), 7.59 (t, J = 7.8 Hz, 1H), 7.35-7.32 (m, 3H), 3.85-3.80 (m, 2H), 3.54-3.51 (m, 2H), 2.55 (t, J = 11.4 Hz, 2H), 1.30 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 160.70, 148.78, 146.45, 144.93, 144.88, 140.94, 138.20, 136.93, 136.16, 135.48, 131.78, 127.24, 127.05, 123.51, 123.00, 121.36, 120.50, 71.39, 71.39 19.01;

HRMS (EI) m / z calcd for C ₂₆ H ₂₄ F ₃ N ₄ O ₃ [M ⁺ ] 497.1801, found: 497.1790.

Example 22: (Compound 22) 3- (6- (4 -methylpiperazin- 1 - yl) pyridin-3-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 ( Preparation of 3H) -one

(4- (trifluoromethoxy) phenyl) methanediol in place of phenylmethanediol in step 1, and 6- (4-methylpiperidin-1-yl) in place of 4-morpholinoaniline in step 2 A pure compound was obtained in the same manner as in Example 10, except that pyridin-3-amine was used.

White solid (5%);

mp 156.4-157.0 ° C .;

¹ H-NMR (400 Mz, CDCl ₃ ) δ 8.42 (dd, J = 1.0, 8.2 Hz, 1H), 8.13 (d, J = 2.8 Hz, 1H), 8.09 (s, 1H), 7.80 (dd, J = 1.2, 7.2 Hz, 1H), 7.67-7.64 (m, 2H), 7.61 (t, J = 7.6 Hz, 1H), 7.54 (dd, J = 2.6, 9.4 Hz, 1H), 7.34 (d, J = 8.4 Hz, 2H), 6.76 (d, J = 8.8 Hz, 1H), 4.35 (d, J = 13.2 Hz, 2H), 2.95-2.88 (m, 2H), 1.76-1.64 (m, 3H), 1.28-1.18 (m, 3H), 0.99 (d, J = 6.0 Hz, 3H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.20, 158.90, 145.88, 145.20, 145.11, 138.25, 136.86, 135.97, 135.42, 131.72, 127.39, 127.08, 123.15, 122.95, 120.55, 106.65, 45.58, 33.73, 33.73 21.90;

HRMS (EI) m / z calcd for C ₂₆ H ₂₄ F ₃ N ₄ O ₂ [M ⁺ ] 481.1851, found: 481.1875.

Example 23: (Compound 23) 3- (6-(( 2S, 6R ) -2,6- dimethylmorpholino ) pyridin-4-yl ) -8- (4- (trifluoromethoxy) phenyl) quina Preparation of Zolin-4 (3H) -one

(4- (trifluoromethoxy) phenyl) methanediol is used instead of phenylmethanediol in step 1, and 6-((2S, 6R) -2,6-dimethyl instead of 4-morpholinoaniline in step 2 Except for using morpholino) pyrimidin-4-amine was prepared in the same manner as in Example 10 to obtain a pure compound.

White solid (12 mg, 6%);

mp 231.9232.7 ° C;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.81 (s, 1H), 8.59 (s, 1H), 8.41 (dd, J = 1.2, 8.4 Hz, 1H), 7.82 (dd, J = 1.2, 7.6 Hz, 1H), 7.667.60 (m, 3H), 7.347.31 (m, 3H), 4.454.19 (m, 2H), 3.713.66 (m, 2H), 3.70 (t, J = 11.8 Hz, 2H) , 1.29 (d, J = 6.4 Hz, 6H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 162.60, 160.63, 158.26, 155.72, 148.86, 144.29, 143.41, 138.37, 136.55, 135.92, 131.77, 127.60, 127.07, 122.66, 120.51, 98.04, 71.47, 49.42, 18.42

HRMS (EI) m / z calcd for C ₂₅ H ₂₃ F ₃ N ₅ O ₃ [M ⁺ ] 498.1753, found: 498.1750.

Example 24 (Compound 24) 3- (6- (4 -Methylpiperidin- 1 - yl) pyrimidin-4-yl) -8- (4 (trifluoromethoxy) phenyl) quinazolin-4 (3H ) -On Preparation

(4- (trifluoromethoxy) phenyl) methanediol in place of phenylmethanediol in step 1, and 6- (4-methylpiperidin-1-yl) pyriin in place of 4-morpholinoaniline in step 2 A pure compound was obtained in the same manner as in Example 10, except that midin-4-amine was used.

White solid (18 mg, 9%);

mp 201.5202.7 ° C;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.77 (s, 1H), 8.56 (s, 1H), 8.41 (dd, J = 1.2, 8.0 Hz, 1H), 7.81 (dd, J = 1.2, 7.2 Hz, 1H), 7.667.58 (m, 3H), 7.33 (d, J = 8.4 Hz, 2H), 7.27 (d, J = 3.6 Hz, 6H), 4.634.29 (m, 2H), 2.97 (t, J = 11.8 Hz, 2H), 1.791.68 (m, 3H), 1.261.16 (m, 2H), 0.99 (d, J = 6.0 Hz, 3H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 162.45, 160.55, 158.40, 155.57, 144.36, 143.64, 138.30, 136.64, 135.78, 131.78, 127.49, 127.09, 122.77, 120.50, 98.02, 44.78, 33.70, 31.10, 21.10

HRMS (EI) m / z calcd for C ₂₅ H ₂₃ F ₃ N ₅ O ₂ [M ⁺ ] 482.1804, found: 482.1809.

Example 25 Preparation of (Compound 25) 3- (2 -morpholinomethyl ) -8- (4- ( trifluoromethoxy ) phenyl) quinazolin-4 (3H) -one

Except for using (4- (trifluoromethoxy) phenyl) methanediol instead of phenylmethanediol in step 1 and 2-morpholinoethanamine instead of 4-morpholinoaniline in step 2, Prepared in the same manner as in Example 10, to obtain a pure compound.

White solid (39%);

mp 124.2125.7 ° C;

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.37 (dd, J = 1.6, 8.0 Hz, 1H), 8.06 (s, 1H), 7.76 (dd, J = 1.6, 7.2 Hz, 1H), 7.647.62 ( m, 2H), 7.57 (d, J = 7.8 Hz, 1H), 7.33 (d, J = 8.4 Hz, 2H), 4.10 (t, J = 6.0 Hz, 2H), 3.67 (t, J = 4.6 Hz, 4H), 2.73 (t, J = 6.0 Hz, 2H), 2.52 (t, J = 4.8 Hz, 4H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.14, 148.77, 146.58, 145.36, 138.02, 136.98, 135.14, 131.69, 126.95, 126.61, 122.74, 120.50, 66.85, 57.03, 53.73, 43.48;

HRMS (EI) m / z calcd for C ₂₁ H ₂₁ F ₃ N ₃ O ₃ [M ⁺ ] 420.1535, found: 420.1552.

Example 26: (Compound 26) of 3- (3- (1H-imidazol-1-yl) propyl) -8- (4- ( pyrofluoromethoxy ) phenyl) quinazolin-4 (3H) -one Produce

(4- (trifluoromethoxy) phenyl) methanediol is used instead of phenylmethanediol in step 1, and 3- (1H-imidazol-1-yl) propane- is used in place of 4-morpholinoaniline in step 2 Except for using 1-amine, to prepare in the same manner as in Example 10 to obtain a pure compound.

Colorless viscous oil (41 mg, 29%).

¹ H-NMR (400Mz, CDCl ₃ ) δ 8.36 (dd, J = 1.4, 7.8 Hz, 1H), 7.94 (s, 1H), 7.78 (dd, J = 1.6, 7.6 Hz, 1H), 7.627.57 ( m, 4H), 7.32 (d, J = 8.0 Hz, 2H), 7.10 (s, 1H), 6.99 (s, 1H), 4.09 (t, J = 7.2 Hz, 2H), 4.01 (t, J = 7.0 Hz, 2H), 2.352.31 (m, 2H).

¹³ C-NMR (CDCl ₃ , 100 MHz) δ 161.22, 148.83, 145.44, 145.21, 138.25, 137.00, 136.70, 135.35, 131.70, 129.72, 127.41, 126.56, 122.58, 120.47, 118.58, 44.26, 44.19, 30.63

HRMS (EI) m / z calcd for C ₂₁ H ₁₈ F ₃ N ₄ O ₂ [M ⁺ ] 415.1382, found: 415.1399.

Example  27: Hedgehog (Hedgehog) Signaling Inhibitory ability  Confirm

Experiments were performed as follows to determine whether the compounds of the present invention inhibit hedgehog signal transduction.

Cell-based assays were performed on NIH3T3 cells containing reporter plasmid (8xGliBS-luciferase) with luciferase bound to the Gli binding site, which is a specific nucleotide sequence where Gli, the final transcription factor of Hedgehog signaling, binds for gene expression. 4x10 ⁴ 8xGliBS-luciferase NIH3T3 cells were cultured in 24 well plates, and then cultured for 30 hours in serum-free DMEM culture the following day to induce cilia formation. After cilia were formed, the hedgehog signal-inducing ligand, Sonic Hedgehog (Shh) -conditioned media, was treated with the compound and the activity was measured using the Dual-luciferase reporter assay system after 24 hours.

As a result, as shown in FIGS. 2 and 3, it was observed that 26 compounds inhibit Hedgehog signal transduction, in particular, 3- (6-((2S, 6R) -2,6-dimethyl Morpholino) pyridin-3-yl) -8-phenylquinazolin-4 (3H) -one (Compound 14) and 3- (6-((2S, 6R) -2,6-dimethylmorpholino) of It was confirmed that the IC ₅₀ concentration of pyridin-3-yl) -8- (4-fluorophenyl) quinazolin-4 (3H) -one (Compound 20) was the lowest.

Accordingly, the present inventors have described the two compounds, 3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8-phenylquinazolin-4 (3H) -one ( See Example 14) and 3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4-fluorophenyl) quinazolin-4 (3H) The following experiment was carried out on -ON (see Example 20).

Example  28: Gli1  Expression Inhibitory ability  Confirm

In order to determine whether the compounds of the present invention inhibit the expression of Gli1, the experiment was performed as follows.

Vismodegib, Sonidegib, Compound 14 (3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8-phenylquinazolin-4 (3H) -one) and Compound 20 (3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4-fluorophenyl) quinazolin-4 (3H) -one) compound Hedgehog signaling activity was measured in 8xGliBS luciferase reporter NIH3T3 as in Example 27 at 7 concentration conditions (0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10 μM).

In addition, Ptch1 ^-/- dermal fibroblasts derived from mice from which the hedgehog signaling inhibitory gene Ptch1 was removed to demonstrate that the hedgehog activity-inhibiting effect of Compound 14 and Compound 20 occur specifically in hedgehog signaling. Experiments were performed using mouse embryonic fibroblasts (MEFs). Ptch1 ^-/- dermal fibroblasts were treated with Vismodegib, Sonidegib, Compound 14 and Compound 20 (10 μM each) for 24 hours in cells where hedgehog signaling activity is always occurring even without Hhh as a ligand. The amount of Gli1 protein whose expression was increased by signaling activity was measured by western blotting using Gli1-specific antibodies.

As a result, as shown in Figures 4 and 5, it was confirmed that Compound 14 and Compound 20 inhibits Gli expression, which indicates that the quinazolinone derivative specifically acts on hedgehog signaling Means that.

Example  29: SMO  protein Inhibitory ability  Confirm

In order to determine whether the compounds of the present invention inhibit the activity of the SMO protein was carried out as follows.

Inhibition of Hedgehog Signaling of Compounds Shh treatment in NIH3T3 cells overexpressing Smo-EGFP to elucidate the molecular mechanism increases the transport of Smo-EGFP proteins to ciliary organelles. At this time, it is known that treatment of Vismodegib and Sonidegib inhibits this process and does not hedge signal transmission.

Thus, Smo-EGFP NIH3T3 cells were treated with serum-free DMEM to induce cilia formation, and then treated with hedgehog ligands and compounds (Vismodegib, Sonidegib, Compound 14 and Compound 20) simultaneously. After 24 hours of compound treatment, the cells were fixed with fixed solution (4% PFA), followed by fluorescence staining using an acetylated-tubulin (Ac-Tub) antibody that labeled the cilia, and then the degree of inhibition of migration to the cilia by Smo-EGFP was compared. It was.

As a result, as shown in Figures 6 and 7, it was confirmed that Compound 14 and Compound 20 inhibited the cilia migration of Smo-EGFP, suggesting that it inhibits SMO protein activity better than Vismodegib, a positive control.

Example  30: Gli2  Expression Inhibitory ability  Confirm

Experiments were carried out as follows to determine whether the compounds of the present invention inhibit the expression of Gli2.

Inhibition of Hedgehog Signaling of Compounds Shh treatment in NIH3T3 cells to determine the molecular mechanism increases the transfer of Gli2 protein, the final signaling factor, to the ciliocyte organelles and increases transcriptional activity.

Therefore, in order to confirm the inhibitory ability of Gli2 expression of the compound of the present invention, after treatment with serum-free DMEM in NIH3T3 cells to induce cilia formation, hedgehog ligands and compounds (Vismodegib, Sonidegib, Compound 14 and Compound 20) simultaneously Treated. After 24 hours of compound treatment, the cells were fixed with fixed solution (4% PFA), followed by fluorescence staining using acetylated-tubulin (Ac-Tub) antibody and Gli2 antibody to label the cilia, and then the degree of migration of Gli2 to the cilia. It was.

As a result, as shown in FIG. 8, it was confirmed that Compound 14 and Compound 20 inhibit the migration of Gli 2 to the cilia.

Example  31: SMO To variant (D477H)  Suppression assessment

Experiments were performed as follows to determine whether the compounds of the present invention inhibit the activity against Smo mutants (D477H) that are resistant to Vismodegib drugs that target first-generation hedgehog signaling.

Shh-conditioned media and 5 different concentrations of Vismodegib, Sonidegib, Compound 14, and Compound 20 were treated for 24 hours in NIH3T3 cells expressing Smo (D477H) mutations that induce steady-state Smo or Vismodegib drug resistance. Gli1 mRNA expression was measured using a quantitative real-time PCR method to determine the degree of hedgehog signal inhibition by drug-resistant Smo mutant (Smo (D477H)).

As a result, as shown in Figure 9, Compound 14 shows that the effect on the SMO variant is not lost.

The foregoing description of the present invention is intended for illustration, and it will be understood by those skilled in the art that the present invention may be easily modified in other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive.

The present invention relates to a novel compound having an SMO inhibitory activity, wherein the isoindolinone derivative and the quinazolinone derivative compound have excellent anticancer effects through the regulation of the hedgehog signaling pathway and the inhibitory activity of the SMO activity. It has been confirmed that it can be usefully used in the field of treatment of cancer diseases.

Claims (14)

A compound represented by Formula 1, or a pharmacologically acceptable salt thereof: [Formula 1]

In Chemical Formula 1, R1 is

,

or

ego; R2 is

or

to be. A compound represented by formula (2), or a pharmacologically acceptable salt thereof: [Formula 2]

In Chemical Formula 2, R1 is Cl,

,

,

,

,

,

,

or

ego; R2 is

,

,

,

,

,

,

,

,

or

to be. The method of claim 1, The compound represented by Formula 1 is a compound represented by Formula 1, characterized in that the compound selected from the group consisting of: 2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -4- (4- (trifluoromethoxy) phenyl) isoindolin-1-one; 2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -4- (pyridin-3-yl) isoindolin-1-one; 4- (2,4-difluorophenyl) -2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) isoindolin-1-one; And 2- (6-((2S, 6R) -2,6-dimethylmorpholino) pyrimidin-4-yl) -4- (4- (trifluoromethoxy) phenyl) isoindolin-1-one. The method of claim 2, The compound represented by Formula 2 is a compound represented by Formula 2, characterized in that the compound selected from the group consisting of: 8-chloro-3- (4-fluorophenyl) quinazolin-4 (3H) -one; 8-chloro-3- (4-morpholinophenyl) quinazolin-4 (3H) -one; 8-chloro-3- (6-morpholinopyridin-3-yl) quinazolin-4 (3H) -one; 8-chloro-3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) quinazolin-4 (3H) -one; 8-chloro-3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyrimidin-4-yl) quinazolin-4 (3H) -one; 3- (4-morpholinophenyl) -8-phenylquinazolin-4 (3H) -one; 8- (3-methoxyphenyl) -3- (4-morpholinophenyl) quinazolin-4 (3H) -one; 3- (4-morpholinophenyl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one; 3- (6-morpholinopyridin-3-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one; 3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8-phenylquinazolin-4 (3H) -one; 3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one ; 3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (naphthalen-1-yl) quinazolin-4 (3H) -one; 3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8-p-tolylquinazolin-4 (3H) -one; 3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4-methoxyphenyl) quinazolin-4 (3H) -one; 3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4- (methylsulfonyl) phenyl) quinazolin-4 (3H) -one; 3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyridin-3-yl) -8- (4-fluorophenyl) quinazolin-4 (3H) -one; 3- (5-((2S, 6R) -2,6-dimethylmorpholino) pyridin-2-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one ; 3- (6- (4-methylpiperazin-1-yl) pyridin-3-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one; 3- (6-((2S, 6R) -2,6-dimethylmorpholino) pyrimidin-4-yl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H)- On; 3- (6- (4-methylpiperidin-1-yl) pyrimidin-4-yl) -8- (4 (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one; 3- (2-morpholinoethyl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one; And 3- (3- (1H-imidazol-1-yl) propyl) -8- (4- (trifluoromethoxy) phenyl) quinazolin-4 (3H) -one. A pharmaceutical composition for preventing or treating cancer, comprising as an active ingredient the compound of any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier. The method of claim 5, The cancer is basal cell carcinoma, medulloblastoma, rhabdomyosarcoma, chondroma, melanoma, small cell lung cancer, non-small cell lung cancer, B-cell lymphoma, multiple myeloma, brain cancer, esophageal cancer, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, liver cancer , Kidney cancer, head and neck cancer, mesothelioma, soft tissue sarcoma, osteosarcoma, testicular cancer, prostate cancer, pancreatic cancer, bone cancer, bone metastasis, acute leukemia, chronic leukemia, glioma, Hodgkins disease, cutaneous melanoma, bladder cancer, endocrine cancer, parathyroid cancer Pharmaceutical composition, characterized in that selected from the group consisting of thyroid cancer, cervical cancer, endometrial cancer, ovarian cancer, skin cancer, renal cell carcinoma, pituitary adenoma, spinal axis tumor, uterine cancer, stomach cancer and biliary tract cancer. The method of claim 5, The compound is a pharmaceutical composition, characterized in that to inhibit the hedgehog signaling pathway (Hedgehog signaling pathway). The method of claim 5, The compound is a pharmaceutical composition, characterized in that to inhibit the SMO (Smoothened) activity. The method of claim 5, The compound is a pharmaceutical composition, characterized in that to inhibit Gli1 expression. The method of claim 5, The compound is a pharmaceutical composition, characterized in that it inhibits the hedgehog signaling pathway (smoothened) variant (D477H) for (Hedgehog signaling pathway). A method of treating a cancer disease comprising administering to a subject a pharmaceutical composition comprising a compound represented by Formula 1, or a pharmacologically acceptable salt thereof: [Formula 1]

In Chemical Formula 1, R1 is

,

or

ego; R2 is

or

to be. A method for treating a cancer disease comprising administering to a subject a pharmaceutical composition comprising a compound represented by Formula 2, or a pharmacologically acceptable salt thereof: [Formula 2]

In Chemical Formula 2, R1 is Cl,

,

,

,

,

,

,

or

ego; R2 is

,

,

,

,

,

,

,

,

or

to be. Use for the treatment of cancer diseases of the pharmaceutical composition comprising a compound represented by the following formula (1), or a pharmacologically acceptable salt thereof: [Formula 1]

In Chemical Formula 1, R1 is

,

or

ego; R2 is

or

to be. Use for the treatment of cancer diseases of the pharmaceutical composition comprising a compound represented by the formula (2), or a pharmacologically acceptable salt thereof: [Formula 2]

In Chemical Formula 2, R1 is Cl,

,

,

,

,

,

,

or

ego; R2 is

,

,

,

,

,

,

,

,

or

to be.

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