KR102114197B1 - Novel benzylideneacetone derivatives and uses thereof - Google Patents

Abstract

The present invention relates to a novel Benzylideneacetone derivative or a use thereof, and more specifically, to a pharmaceutical composition for preventing or treating cancer or bone disease, including the novel Benzylideneacetone derivative or a pharmaceutically acceptable salt thereof as an active ingredient, improvement It is for a food composition.
Since the novel Benzylideneacetone derivatives according to the present invention show strong proliferation and differentiation inhibitory activity against osteoclasts that cause bone loss in addition to cancer cell specific cytotoxicity, safe and effective anticancer drugs and osteoporosis prevent and treat or improve bone diseases It can be usefully used to develop food products.

Description

Novel benzylideneacetone derivatives and uses thereof

The present invention relates to a novel benzylidene acetone (Benzylideneacetone) derivative or a use thereof, more specifically for the prevention or treatment of cancer or bone disease comprising the novel Benzylideneacetone derivative or a pharmaceutically acceptable salt thereof as an active ingredient It relates to a pharmaceutical composition, a composition for improving food.

Cancer is the leading cause of death in the aging society. Currently, with the development of surgical surgery, chemotherapy, and radiation therapy, the cure rate is over 70% if all types of cancer are included in early and late stages. However, due to the technical limitations of surgical surgery and the limited doses due to the side effects of chemotherapy and radiotherapy, it is difficult to cure cancer in terminal cancer, metastatic cancer, and recurrent cancer, and the mortality rate is very high. In addition, patients who are unable to chemotherapy due to the expression of resistance to existing anticancer drugs are increasing.

Therefore, the therapeutic effect is stronger, the side effects are very low and safe, so it is possible to increase the cancer cure rate by high-dose administration, and there is an urgent need to develop a new anti-cancer agent that has a therapeutic effect even in patients with resistance to existing anticancer drugs.

Bones play a very important role in forming the body's skeletal structure and maintaining calcium (Ca ²⁺ ) levels in the blood. Bone is maintained through a balance of bone remodeling cycles between metabolically absorbing osteoclasts and producing osteoblasts. When the balance between bone absorption and production is destroyed and the amount of absorption is greater than the amount of production, various bone-related diseases occur. Representative diseases related to osteoclast differentiation and activation are osteoporosis, rheumatoid arthritis, joint pain, Paget's disease, bone metastasis cancer, and Fractures, etc. (Kim JH and Kim N, 2016; Shiozawa Y et al ., 2011; Singer FR, 2016).

Rheumatoid arthritis is an autoimmune disease, but autoimmune antibodies promote osteoclast differentiation. The resulting excessive bone absorption exacerbates rheumatoid arthritis (Takayanagi H, 2007). The mechanism is as follows. NFAT transcription factors (NFATc1 / c2 / c3 / c4), a central transcription factor related to osteoclast differentiation, are basically activated by calcium / calmodulin signaling (Takayanagi H et al ., 2002). For complete activation, tyrosine-based activation motif (ITAM) -bearing molecules such as the immunomodulatory proteins DNAX-activating protein 12 (DAP12) and the immune antibody Fc receptor common γ chain (FcRγ) stimulate calcium signaling in immune cells ( Pitcher LA and van Oers NS, 2003). In osteoclasts, DAP12 and FcRγ also activate NFATc1 through calcium signaling. Therefore, immunoglobulin-like receptors linked to DAP12 and FcRγ play an important role in osteoclast differentiation (Koga T et al ., 2004; MoA et al ., 2004). That is, FcRγ interacts with osteoclast-associated receptor (OSCAR) and paired immunoglobulin-like receptor (PIR-A) in osteoclasts. When ITAM is phosphorylated, it activates phospholipase C γ (PLCγ), which promotes intracellular calcium, which activates calmodulin-dependent phosphatase, calcineurin. Calcineurin dephosphorylates serine of NFATc1 directly into the nucleus and activates it. As a result, immune antibodies promote osteoclast differentiation, and excessive bone absorption by osteoclasts exacerbates rheumatoid arthritis. Eventually, suppression of osteoclast differentiation in rheumatoid arthritis patients can treat skeletal symptoms such as arthritis and pain caused as a result of not correcting the abnormalities of the autoimmune mechanism itself.

Osteitis deformans also causes abnormal bone resorption of osteoclasts (Singer FR, 2016). Then, the abnormal bone formation of osteoblasts progresses and this process is repeated, resulting in bone deformity and resulting pain, headache, and hearing loss. It affects the arms, legs, pelvis, spine and skull. The newly created bones are weak and the frequency of fractures is high. Hypercalcemia, heart attack and hemiplegia can be induced (Ralstone SH, 2016). The cause of the outbreak is unknown, but genetic predisposition and childhood viral infections are suspected. Medication can help suppress disease progression. Currently, the most used treatments are Fosamax, an inhibitor of osteoclast differentiation, and calcitonin, which regulates bone metabolism. However, Fosamax is a side effect and long-term use is limited in some patients. For severe pain, use Acetaminophen (Tylenol) or nonsteroidal anti-inflammatory drugs (NSAIDs).

Osteoclasts also promote bone metastasis of solid tumors. Bone is the most prone to cancer metastasis. When cancer is metastasized to the bone, the likelihood of cure is significantly lowered due to severe bone pain and fracture (Weilbaecher KN et al ., 2011). Cancer cells spread throughout the body are found at the site of proliferation of blood stem cells in the bone marrow (Shiozawa Y et al ., 2013). Cancer cells significantly promote the differentiation of osteoclasts from bone marrow cells, thereby promoting bone destruction, resulting bone metastasis, and cancer growth. Therefore, osteoclasts play a key role in bone metastasis of cancer, and when osteoclast differentiation is suppressed, bone metastasis decreases. The metastasis of many solid cancers is bone metastasis, and blood stem cells are driven out from the place where blood stem cells proliferate, and then they are multiplied and then released into the blood before being transferred to other places. The most common cancer of bone metastasis is prostate cancer, and bone metastasis exacerbates cancer, making it difficult to cure and leading to death. The main direct target of human prostate cancer cells is also used as a base for metastatic cancer as a site of proliferation of blood stem cells (Shiozawa Y et al ., 2011). In addition, osteoclasts promote blood vessel formation in prostate cancer tissues, thereby promoting cancer growth (Bruni-Cardoso A et al ., 2010). Breast cancer cells also promote osteoclast differentiation, where osteoclasts promote cancer recurrence through bone metastasis in breast resection patients (Danilin S et al ., 2012; Lu X et al ., 2011).

Bone targeting therapies to prevent bone metastasis are currently being used in clinical trials, and osteoclasts are one of the key mechanisms of cancer metastasis, making them a major target for the development of new anticancer drugs. Accordingly, the only bisphosphonate-based drug currently approved by the United States FDA for the purpose of suppressing osteoclast differentiation is Zoledronic acid (El-Amm J et al ., 2013). Zoledronic acid preserves bones and increases survival. Zoledronic acid significantly reduced bone metastasis in high risk nonmetastatic prostate cancer (Wirth M et al ., 2014). Bone metastasis was further reduced when zoledronic acid was administered with parathyroid hormone, an agent that promotes bone formation (Schneider A et al ., 2005). Denosumab, a monoclonal antibody against RANKL, a signaling material for osteoclast differentiation, also inhibited bone metastasis of prostate cancer, and it was again proved that osteoclast suppression is important for suppressing bone metastasis of cancer (Smith MR et al . , 2012). In multiple myeloma patients, administration of Zoledronic acid inhibited osteoclast differentiation, significantly inhibiting bone metastasis (Zhuang J et al ., 2012).

Osteoporosis is caused when the balance between bone absorption and production is destroyed by the activation of osteoclasts and the absorption amount is greater than the production amount. Osteoporosis increases the frequency of fractures by reducing the density of the bone parenchyma. It occurs most frequently in women who have a hormonal balance disorder, such as middle-aged and older women, and occurs in patients who are unable to behave due to a fracture or severe disease. Recently, the incidence has increased in men over the age of middle-aged.

In the molecular mechanism by which bone marrow monocyte / macrophage lineage cells differentiate into osteoclasts, the following two cytokines play an important role (Teitelbaum SL and Ross FP, 2003). (i) When macrophage colony-stimulating factor (M-CSF) is combined with its receptor, c-Fms, osteoclast progenitor cells proliferate and survive. Receptor activator of nuclear factor-κB ligand (RANKL) binds to its receptor, RANK, which activates osteoclast differentiation and bone resorption and allows mature osteoclasts to survive (Lacey DL et al ., 1998; Lum L et al. ., 1999; Sherr CJ, 1990; Suda T et al ., 1999; Wong BR et al ., 1999). (ii) When M-CSF induces activation of c-Fms, osteoclast progenitor cells proliferate and survive through ERK and PI3K / Akt pathways (Mancini et al ., 1997). (iii) RANKL (OPGL, ODF, TRANCE) and RANK also regulate osteoclast formation and function (Anderson DM et al ., 1997; Dougall WC et al ., 1999; Kong YY et al ., 1999). When RANKL binds to RANK, TNF receptor-associated factors (TRAFs) such as TRAFs 1, 2, 3, 5, and 6 bind RANK (Darnay BG et al ., 1998; Walsh MC and Choi Y, 2003). Of these, TRAF6 is most important for osteoclast formation and function (Lomaga MA et al ., 1999; Naito A et al ., 1999). TRAF6 delivers RANKL / RANK signals to NF-κB, c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), p38, Akt, and Nuclear Factor Of Activated T-Cells 1 (NFATc1). Cell proliferation, fusion, and differentiation are achieved (Kobayashi N et al. , 2001; Lomaga MA et al ., 1999; Naito A et al ., 1999; Takayanagi H et al ., 2002; Wong BR et al ., 1998; Wong BR et al ., 1999)

The existing direction of developing a therapeutic agent for osteoporosis was to discover substances capable of preventing bone loss by inhibiting osteoclast bone absorption. The representative drug is bisphosphonate-based Fosamax. In the same vein, many studies have been conducted on the effects of arachidonate metabolites on bone tissue metabolism (Lee Sung-eun, 1999). Leukotriene-B4 (LTB4) is one of the metabolites of the 5-lipoxygenase pathway, a metabolic pathway of arachidonate (Ford-Hutchinson, AW et al ., 1980). C433, an interstitial cell obtained from a giant cell tumor, has been reported to increase the number and activity of osteoblasts by increasing 5-lipoxygenase metabolites (Mundy, GR et al. , 1993). In the bone tissue culture process, it was observed that when LTB4 was administered, bone absorption increased (Bonewald, LF et al. , 1996). In vitro and in vivo studies have also shown that LTB4 induces bone resorption by increasing osteoclast production (Bonewald, LF et al. , 1996). Accordingly, LTB4 receptor antagonists (LTB4 receptor antagonists) have been developed a lot to treat osteoporosis, but did not succeed in sufficiently suppressing the bone parenchymal absorption of osteoclasts.

In addition, the side effects of the existing treatments for osteoporosis and expensive drug prices have also become a major obstacle to administration in sufficient doses for treatment of patients. The main side effects of Fosamax include severe esophagitis, kidney damage, liver damage, hypocalcemia, and muscle spasms. Roche's Bonviva has side effects such as generalized muscle pain and body aches. Novartis' Aclasta (zoledronate) and Eli Lilly's anabolic drugs parathyroid hormone Forsteo and Forteo (teriparatide) have good effects, but are too expensive to limit their use. In particular, Forsteo / Forteo is active in metabolic bone diseases such as hypersensitivity patients, pregnant women, breastfeeding, hypercalcemia, renal failure, hyperparathyroidism and Paget'disease, and unknown causes of alkaline phosphatase. Because it cannot be used in patients with unexplained elevations of alkaline phosphatase, radiation therapy, bone marrow cancer or bone metastasis, the applicable patient population is not large.

Therefore, there is an urgent need to develop a treatment for bone-related diseases that is more effective, safe without side effects, and can be produced at a lower cost than existing treatments.

[Non-Patent Document 1] Anderson DM, Maraskovsky E, Billingsley WL, Dougall WC, Tometsko ME, Roux ER, et al. A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature 1997; 390: 175-9. [Non-Patent Document 2] Bonewald, L. F., et al., Leukotriene B4 stimulates osteoclastic bone resorption both in vitro and in vivo. J. Bone Miner. Res., 11, 1619-1627, 1996. [Non-Patent Document 3] Bonewald, L.F. et al., Effects of synthetic peptido-leukotrienes on bone resorption in vitro. J. Bone Miner. Res., 11, 521-529, 1996.

Accordingly, the present inventors synthesized a novel Benzylideneacetone derivative to develop a therapeutic agent for cancer or bone disease with little side effects, safety, and excellent effect, and thus completed the present invention by confirming that it has anti-cancer activity and bone loss-inhibiting activity.

Therefore, the object of the present invention

To provide a Benzylideneacetone derivative represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

<Formula 1>

[In the above formula, R1 and R2 are different,

R1 and R2 are each independently -H, -OH, -SH, C _1-4 linear or branched alkyl group, C _1-4 linear or branched alkoxy group, halogen, allyloxy group, allyloxy, benzyloxy group ( benzyloxy), an aryloxy group having one or more selected from the group consisting of heteroatoms and substituents, and a heterocycloalkyl group having 3-7 atoms with one or more heteroatoms, wherein the hetero The atom consists of O, N and S, respectively:

R3 is a straight or branched alkyl group of C _1-4 , -NHR4, -N (R4) ₂ , OH, respectively:

R4 are each independently a C _1-4 straight or branched chain alkyl group.]

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer comprising the Benzylideneacetone derivative represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating bone disease, which includes the Benzylideneacetone derivative represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a method for producing a Benzylideneacetone derivative of claim 1.

In order to achieve the above object,

The present invention provides a Benzylideneacetone derivative represented by Formula 1 below or a pharmaceutically acceptable salt thereof.

<Formula 1>

[In the above formula, R1 and R2 are different,

R1 and R2 are each independently -H, -OH, -SH, C _1-4 linear or branched alkyl group, C _1-4 linear or branched alkoxy group, halogen, allyloxy group, allyloxy, benzyloxy group ( benzyloxy), aryloxy having one or more selected from the group consisting of heteroatoms and substituents, and one selected from the group consisting of 3-7 membered heterocycloalkyl groups having one or more heteroatoms, wherein the heteroatoms Is composed of O, N and S, respectively:

R3 is a straight or branched alkyl group of C _1-4 , -NHR4, -N (R4) ₂ , OH, respectively:

R4 are each independently a C _1-4 straight or branched chain alkyl group.]

In order to achieve another object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating cancer comprising Benzylideneacetone derivative represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

<Formula 1>

[In the above formula, R1 and R2 are different,

R1 and R2 are each independently -H, -OH, -SH, C _1-4 linear or branched alkyl group, C _1-4 linear or branched alkoxy group, halogen, allyloxy group, allyloxy, benzyloxy group ( benzyloxy), an aryloxy group having one or more selected from the group consisting of heteroatoms and substituents, and a heterocycloalkyl group having 3-7 atoms with one or more heteroatoms, wherein the hetero The atom consists of O, N and S, respectively:

R3 is a straight or branched alkyl group of C _1-4 , -NHR4, -N (R4) ₂ , OH, respectively:

R4 are each independently a C _1-4 straight or branched chain alkyl group.]

In order to achieve another object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating bone disease comprising Benzylideneacetone derivative represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

<Formula 1>

[In the above formula, R1 and R2 are different,

R1 and R2 are each independently -H, -OH, -SH, C _1-4 linear or branched alkyl group, C _1-4 linear or branched alkoxy group, halogen, allyloxy group, allyloxy, benzyloxy group ( benzyloxy), an aryloxy group having at least one selected from the group consisting of heteroatoms and substituents, and a heterocycloalkyl group of 3-7 atoms having one or more heteroatoms, wherein the hetero The atom consists of O, N and S, respectively:

R3 is a straight or branched alkyl group of C _1-4 , -NHR4, -N (R4) ₂ , OH, respectively:

R4 are each independently a C _1-4 straight or branched chain alkyl group.]

In order to achieve another object of the present invention, the present invention provides a method for producing a Benzylideneacetone derivative of claim 1.

Hereinafter, the present invention will be described in detail.

The present invention provides a Benzylideneacetone derivative represented by Formula 1 below or a pharmaceutically acceptable salt thereof.

<Formula 1>

[In the above formula, R1 and R2 are different,

R1 and R2 are each independently -H, -OH, -SH, C _1-4 linear or branched alkyl group, C _1-4 linear or branched alkoxy group, halogen, allyloxy group, allyloxy, benzyloxy group ( benzyloxy), an aryloxy group having at least one selected from the group consisting of heteroatoms and substituents, and a heterocycloalkyl group of 3-7 atoms having one or more heteroatoms, wherein the hetero The atom consists of O, N and S, respectively:

R3 is a straight or branched alkyl group of C _1-4 , -NHR4, -N (R4) ₂ , OH, respectively:

R4 are each independently a C _1-4 straight or branched chain alkyl group.]

<Benzylideneacetone>

Benzylideneacetone is a compound having the structure of the formula above, represented by the molecular formula of C ₁₀ H ₁₀ O (molecular weight 146.19 g / mol) and the above structural formula. Benzalacetone, Methyl Styryl ketone, Benzylidene acetone, or IUPAC name (E) -4-Phenylbut-3-ene-2-one, present as yellow crystals at room temperature.

The novel compounds of the present invention have the following structure as a derivative of the Benzylideneacetone.

<Formula 1>

[In the above formula, R1 and R2 are different,

R1 and R2 are each independently -H, -OH, -SH, C _1-4 linear or branched alkyl group, C _1-4 linear or branched alkoxy group, halogen, allyloxy group, allyloxy, benzyloxy group ( benzyloxy), an aryloxy group having at least one selected from the group consisting of heteroatoms and substituents, and a heterocycloalkyl group of 3-7 atoms having one or more heteroatoms, wherein the hetero The atom consists of O, N and S, respectively:

R3 is a straight or branched alkyl group of C _1-4 , -NHR4, -N (R4) ₂ , OH, respectively:

R4 are each independently a C _1-4 straight or branched chain alkyl group.]

The term “alkyl” used in the present invention is used to describe a group or part of a group comprising a straight or branched chain alkyl group containing 1 to 4 carbon atoms; Examples of such groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert butyl.

The term "alkoxy" used in the present invention means O-alkyl.

The "alkyl" described in the definition of alkoxy is the same as the "alkyl group" used in the present invention, specifically, a methoxy group, ethoxy group, propoxy group, isopropoxy group, n-butoxy group, isobutoxy group, t- It is a butoxy group.

The term “aryloxy” used in the present invention means O-aryl.

"Aryl" described in the definition of aryloxy is the same as "benzyl group" used in the present invention. Specifically, it includes an unsubstituted aryl group or a heteroaryl group containing one or more N, S or O heteroatoms. The aryloxy group containing the unsubstituted aryl group is the same as "benzyloxy" used in the present invention.

In addition, one or more substituents may be independently included as substituents of the aryl group. At this time, the substituent specifically includes a hydroxy group, an amino group, an alkyl group, an alkoxy group, a carboxylic acid group, a carboxyl ester, and a carboxyamide.

The term 'carboxyl ester group' used in the present invention refers to an ester in which O-alkyl or O-aryl group is substituted, and 'carboxyamide group' is N-alkyl, N, N-dialkyl, N-aryl, N, N-diaryl.

The aryl group, alkyl group, and alkoxy group are as described above.

The term 'halogen' used in the present invention represents a halogen group atom, and includes fluorine, chlorine, bromine, and iodine, and the novel derivative of the present invention may preferably be R2 halogen.

"One of the 3-7 membered heterocycloalkyl groups having one or more substituents of the present invention, the substituent is O, N, S" is preferably one having the following structural formula have.

More preferably, the novel derivative of Formula 1 of the present invention is selected from the following.

(1) (E) -4- (4-fluoro-3-hydroxyphenyl) but-3-en-2-one (M.W 176.21; brown solid)

(2) (E) -4- (3-hydroxy-4-methylphenyl) but-3-en-2-one (M.W 176.21; brown solid)

(3) (E) -3- (3-hydroxy-4-methylphenyl) acrylamide (M.W 177.2; yellow solid)

(4) (E) -3- (4-fluoro-3-hydroxyphenyl) acrylamide (M.W 181.17; yellow solid)

(5) (E) -4- (4-mercapto-3-hydroxyphenyl) but-3-en-2-one

(6) (E) -4- (4-hydroxy-3-mercaptophenyl) but-3-en-2-one

(7) (E) -4- (3-hydroxy-4-isopropoxyphenyl) but-3-en-2-one

(8) (E) -4- (4- (benzyloxy) -3-hydroxyphenyl) but-3-en-2-one

(9) (E) -4- (4- (allyloxy) -3-hydroxyphenyl) but-3-en-2-one

(10) (E) -4- (3-hydroxy-4- (4-methylpiperazin-1-yl) phenyl) but-3-en-2-one

(11) (E) -4- (3-hydroxy-4- (pyrrolidin-1-yl) phenyl) but-3-en-2-one

(12) (E) -4- (3-hydroxy-4- (4-hydroxyphenoxy) phenyl) but-3-en-2-one

(13) (E) -4- (3-hydroxy-4- (piperidin-4-yloxy) phenyl) but-3-en-2-one

Benzylideneacetone derivatives included in the composition of the present invention may be used as such or in the form of salts, preferably pharmaceutically acceptable salts. 'Pharmaceutically acceptable' in the present invention refers to a physiologically acceptable, when administered to a human, that usually does not cause an allergic reaction or a similar reaction, and the salt is a pharmaceutically acceptable free acid (free acid). The acid addition salt formed by) is preferred. Organic acids and inorganic acids can be used as the free acid. The organic acid is not limited thereto, citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, metasulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, Glutamic acid and aspartic acid. In addition, the inorganic acids include, but are not limited to, hydrochloric acid, bromic acid, sulfuric acid and phosphoric acid.

Benzylideneacetone derivatives represented by Chemical Formula 1 of the present invention can be naturally isolated or prepared by a chemical synthesis method of a compound known in the art, or by the production method described herein.

In order to achieve another object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating cancer or bone disease, comprising as an active ingredient a Benzylideneacetone derivative represented by Formula 1 or a pharmaceutically acceptable salt thereof.

Benzylideneacetone derivative represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof is as described above.

The derivative of Formula 1 of the present invention is very effective in the treatment of cancer. Examples of the cancer include breast cancer, colorectal cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, and anal anus Cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, Cancers such as prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or urinary tract cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, pituitary adenoma, or of these cancers It can be one or more combinations.

The present inventors confirmed that in one embodiment, Benzylideneacetone derivatives showed cytotoxicity against various cells, and showed strong cytotoxicity in cancer cells but little cytotoxicity in normal cells.

Therefore, a person skilled in the art can understand that by using the above activity of Benzylideneacetone derivatives identified by the present inventors, effective prevention of cancer, improvement of symptoms, or effects of treatment can be expected.

In another embodiment, the present inventors confirmed that Benzylideneacetone derivatives effectively inhibit the differentiation of osteoclasts, which serve to destroy and reabsorb bone tissue. As a result of separating monocytes, which are stem cell progenitor cells of osteoclasts, from bone marrow cells isolated from mice, stimulating them with differentiation promoting factors RANKL and M-CSF, and treating Benzylideneacetone derivatives to determine the effect on osteoclast differentiation, bone marrow Cells were effectively suppressed from being differentiated into multinuclear osteoclasts.

The present inventors have confirmed that Benzylideneacetone derivatives not only have excellent osteoclast differentiation inhibitory activity, but also have very low cytotoxicity and are safe.

Therefore, a person skilled in the art utilizes the above-described activity of Benzylideneacetone derivatives identified by the present inventors, and bone density and strength are broken while the balance between bone absorption by osteoclasts and formation of new bone matrix by osteoblasts and bone metabolism in subsequent mineralization processes is broken. It can be understood that an effective prevention, improvement of symptoms, or treatment effect can be expected for various bone diseases caused by decreasing.

In the present invention, the bone disease is, for example, but not limited to, osteoporosis, Paget's disease, rickets, osteomalacia, renal osteotrophy of patients with renal failure, joint pain, fracture, rheumatoid bone disease, degenerative bone disease, bone metastasis, Primary tumors generated in bone, periodontal disease, inflammatory alveolar bone resorption disease and inflammatory bone resorption disease may be, preferably osteoporosis, Paget's disease, rickets, joint pain, fracture, rheumatoid bone disease, bone metastasis cancer. For the correlation between each disease and osteoclast, refer to the background and examples of the present specification.

'Treatment' as used herein refers to a clinical procedure to alter the natural course of the individual or cell being treated, and may also be performed for the prevention of clinical pathology. The desirable effects of treatment include suppressing the occurrence or recurrence of the disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, reducing the rate of disease progression, improving the condition of the disease, improving, alleviating or improving the prognosis. Includes. The term 'prevention' also means any action that suppresses the onset of disease or delays its progression.

The dosage of the pharmaceutical composition of the present invention includes the route of administration, the time of administration, the number of treatments, the duration of treatment, the age of the individual in need of treatment, weight, health status, sex, the severity of the disease, sensitivity to drugs, diet and excretion rate, Considering various factors such as, one of ordinary skill in the art can determine an appropriate effective amount according to the specific use described above. The “effective amount” refers to an amount sufficient to exhibit an improvement, treatment, prevention, detection or diagnosis effect of cancer or bone disease when administered to an individual. The "individual" may be an animal, preferably a mammal, most preferably an animal including a human, or may be a cell, tissue, or organ derived from an animal. The subject may be a patient with bone disease in need of treatment.

The administration may be administered once or several times a day. The pharmaceutical composition of the present invention may be administered alone or in combination with other therapeutic agents known to be effective in the prevention or treatment of bone disease, and when administered in combination, may be administered sequentially or simultaneously with other therapeutic agents. When administered alone or in combination, the dosage of the pharmaceutical composition of the present invention is preferably administered in an amount that can obtain the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art.

The pharmaceutical composition of the present invention may be variously formulated according to the route of administration by a method known in the art together with a pharmaceutically acceptable carrier. 'Pharmaceutically acceptable' refers to a non-toxic composition that is physiologically acceptable and does not inhibit the action of the active ingredient when administered to humans and does not normally cause an allergic or similar reaction such as gastrointestinal disorders, dizziness, or the like. . The carrier includes all kinds of solvents, dispersion media, oil-in-water or water-in-oil emulsions, aqueous compositions, liposomes, microbeads, and microsomes.

The route of administration may be administered orally or parenterally. The parenteral administration method is not limited thereto, but intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal administration. Can be.

When the pharmaceutical composition of the present invention is orally administered, the pharmaceutical composition of the present invention is a powder, granules, tablets, pills, dragees, capsules, liquids, gels according to methods known in the art together with a suitable carrier for oral administration. , Syrups, suspensions, wafers, and the like. Examples of suitable carriers include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol, and starches including cellulose starch, wheat starch, rice starch and potato starch, cellulose, and the like. Fillers such as cellulose, gelatin, polyvinylpyrrolidone and the like may be included, including methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethylcellulose. In addition, if necessary, cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant. Furthermore, the pharmaceutical composition may further include an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent and a preservative.

In addition, when administered parenterally, the pharmaceutical composition of the present invention may be formulated according to methods known in the art in the form of injections, transdermal administrations, and nasal inhalants together with suitable parenteral carriers. The injections must be sterile and protected from contamination of microorganisms such as bacteria and fungi. For injections, examples of suitable carriers include, but are not limited to, solvents or dispersion media comprising water, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and / or vegetable oils. Can be. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) with ethanol amine or sterile water for injection, isotonic solutions such as 10% ethanol, 40% propylene glycol and 5% dextrose. Etc. can be used. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, etc. may be further included. In addition, the injection may additionally contain isotonic agents such as sugars or sodium chloride in most cases.

In the case of transdermal administration agents, ointments, creams, lotions, gels, external solutions, pasta, linen agents, air rolls, and the like are included. In the above, 'transdermal administration' means that the pharmaceutical composition is topically administered to the skin to deliver an effective amount of the active ingredient contained in the pharmaceutical composition into the skin. For example, the pharmaceutical composition of the present invention can be administered by a method of preparing the injectable formulation and lightly pricking the skin with a 30-gauge thin injection needle or directly applying it to the skin. These formulations are described in Remington's Pharmaceutical Science, 15th Edition, 1975, Mack Publishing Company, Easton, Pennsylvania, a recipe generally known in pharmaceutical chemistry.

In the case of inhalation dosing agents, the compounds used in accordance with the present invention may be pressurized packs or It can be conveniently delivered in the form of an aerosol spray from a nebulizer. In the case of pressurized aerosols, the dosage unit can be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges used in inhalers or insufflators can be formulated to contain a powder mixture of a compound and a suitable powder base such as lactose or starch.

As other pharmaceutically acceptable carriers, reference may be made to those described in the following documents (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).

In addition, the pharmaceutical composition according to the present invention may include one or more buffers (e.g. saline or PBS), carbohydrates (e.g. glucose, mannose, sucrose or dextran), antioxidants, bacteriostatic agents, chelating agents (Eg, EDTA or glutathione), adjuvants (eg, aluminum hydroxide), suspending agents, thickening agents and / or preservatives.

In addition, the pharmaceutical compositions of the present invention can be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.

In addition, the present invention provides a composition for the prevention or improvement of cancer or bone disease comprising Benzylideneacetone derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

The effects of prevention or improvement of the Benzylideneacetone derivatives identified by the present inventors on the cancer or bone disease are as described above.

The food composition includes all forms of functional foods, nutritional supplements, health foods, and food additives. These types can be prepared in various forms according to conventional methods known in the art.

For example, as a health food, the food composition itself of the present invention may be prepared in the form of tea, juice, and drink to be consumed or granulated, encapsulated, and powdered. In addition, the food composition of the present invention can be prepared in the form of a composition by mixing with a known substance or active ingredient known to have an effect of preventing or improving bone disease.

In addition, functional foods include beverages (including alcoholic beverages), fruits and processed foods thereof (e.g. canned fruits, canned fruits, jams, marmalades, etc.), fish, meat and processed foods (e.g. ham, sausage, Corn beef, etc.), breads and noodles (e.g. udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), juice, various drinks, cookies, syrup, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, Margarine, vegetable protein, retort food, frozen food, various seasonings (for example, miso, soy sauce, sauce, etc.) can be prepared by adding the food composition of the present invention.

The preferred content of the food composition according to the present invention is not limited thereto, but is preferably from 0.01 to 50% by weight of the total weight of the food finally produced. In order to use the food composition of the present invention in the form of a food additive, it may be prepared and used in the form of a powder or a concentrate.

The present invention provides a method for producing the Benzylideneacetone derivative of claim 1.

The derivative of claim 1 of the present invention is prepared under a suitable reaction solvent. Solvents that can be used are methanol, ethanol, propanol, butanol, chloroform, dichloromethane, ethyl acetic acid, nucleic acid, benzene, methylene chloride, acetone, tetrahydrofuran (THF), dioxane, DMF or a mixed solvent thereof.

A derivative in which R3 of the derivative of Formula 1 of the present invention is a methyl group may be prepared by Preparation Method 1 or Method 3 below, and a derivative in which R3 is -NH ₂ may be prepared by Method 2 below.

Hereinafter, the manufacturing method according to the present invention will be described in detail step by step.

Method 1 (Method 1)

In the present invention, the following step a is a step of preparing compound 2 by hydrolyzing the methyl ether of compound 1 under acid catalyst conditions.

At this time, BBr ₃ , AlCl ₃ , HBr, etc. may be used as an acid used as a catalyst. At this time, CH ₃ CN, CH ₂ Cl ₂ and the like may be used as the reaction solvent.

Step b below is a step of preparing compound 3 by reacting compound 2 prepared in step a with CuBr ₂ . In step b, an organic solvent such as acetone or DMF may be used as a reaction solvent, and the reaction temperature is preferably performed at 40 ° C to 70 ° C.

[Method 1]

R of the method 1 is -H, -OH, -SH, C _1-4 straight or branched chain alkyl group, C _1-4 straight or branched alkoxy group, halogen, allyloxy group (allyloxy), benzyloxy group (benzyloxy ), An aryloxy group having one or more selected from the group consisting of heteroatoms and substituents, and a heterocycloalkyl group having 3-7 atoms having one or more heteroatoms, wherein the heteroatom is Is composed of O, N and S, respectively.

The alkyl group, alkoxy group, aryloxy, halogen, cycloalkyl and the like are as described in the above specification.

Method 2 (Method 2)

In the present invention,

The following step a 'is a step of preparing compound 2 by reacting compound 1 with triethyl phosphonoacetate. At this time, THF, methanol, water, etc. may be used as the reaction solvent, and the reaction temperature is preferably performed at room temperature.

The following step b 'is a step of preparing compound 3' by reacting a methyl ester group of compound 2 'with a base. At this time, LiOH, NaOH, or the like may be used as the base, and THF, methanol, and water may be used as the reaction solvent.

Step c 'below is a step for preparing compound 4' by reacting the hydroxy group of compound 3 'with NH ₄ HCO ₃ . At this time, dioxane may be preferably used as the reaction solvent.

The following step d 'is a step of preparing the compound 5' by hydrolyzing the methyl ether of the compound 4 'under acid catalyst conditions.

At this time, BBr ₃ , AlCl ₃ , HBr, etc. may be used as an acid used as a catalyst. At this time, CH ₃ CN, CH ₂ Cl ₂ and the like may be used as the reaction solvent.

[Method 2]

R of the method 2 is -H, -OH, -SH, C _1-4 straight or branched alkyl group, C _1-4 straight or branched alkoxy group, halogen, allyloxy group (allyloxy), benzyloxy group (benzyloxy ), An aryloxy group having one or more selected from the group consisting of heteroatoms and substituents, and a heterocycloalkyl group having 3-7 atoms having one or more heteroatoms, wherein the heteroatom is Is composed of O, N and S, respectively.

The alkyl group, alkoxy group, aryloxy, halogen, heterocycloalkyl and the like are as described in the above specification.

Manufacturing method 3 (method 3)

In the present invention, the following step a "is a step of preparing a compound 2" in which a protective group is selectively introduced using a halogenated compound under the basic conditions of compound 1 ". At this time, K ₂ CO ₃ , KHCO ₃ , NaHCO ₃ can be used, and acetone, DMF, etc. can be used as the reaction solvent, and the reaction temperature is preferably performed at 40 to 80 ° C. The halogenated compound is bromobenzene, benzyl bromide, bromopro. Pens, 2-bromopropane, and the like can be used.

The following step b “is a step of preparing compound 3” by reacting compound 2 “prepared in step a” with CuBr ₂ . Step b “may be acetone, DMF or the like as a reaction solvent, and the reaction temperature is preferably performed at 40 ° C to 70 ° C.

[Method 3]

R of the method 3 is C _1-4 straight or branched chain alkyl group, C _1-4 straight or branched alkoxy group, C _1-4 alkenyl group; Aryl group having one or more selected from the group consisting of heteroatoms and substituents and 3-7 membered heterocycloalkyl groups having one or more heteroatoms, wherein the heteroatoms are O, N and S Is made;

The alkyl group, alkoxy group, aryl group, halogen, cycloalkyl and the like are as described in the above specification.

The novel benzylidene acetone derivative according to the present invention is not limited to the above production method, and any known method as well as other known methods can be used as long as it can synthesize the novel benzylidene acetone derivative.

The novel derivatives of the present invention prepared under the above reaction solvent can be separated after production using concentration gradient chromatography. As the chromatography, column chromatography filled with various synthetic resins such as silica gel or activated alumina and high-speed liquid chromatography (HPLC) may be used alone or in combination. In the examples of the present invention, silica gel column chromatography and flash column chromatography were used.

Accordingly, the present invention provides a novel Benzylideneacetone derivative, or a composition for improving or treating cancer or bone disease and a method for manufacturing the same, comprising the pharmaceutically acceptable salt thereof as an active ingredient. The composition according to the present invention has a strong anti-cancer effect and also has a strong proliferation and differentiation inhibitory effect on osteoclasts that cause bone loss, and has very low cytotoxicity against normal cells, and can be usefully used as a therapeutic agent for cancer or bone disease. have.

1 is a result of NMR identification of (E) -4- (4-fluoro-3-hydroxyphenyl) but-3-en-2-one synthesized in Example 1-1.
2 is a result of NMR identification of (E) -4- (3-hydroxy-4-methylphenyl) but-3-en-2-one synthesized in Example 1-2.
3 is a result of NMR identification of (E) -3- (3-hydroxy-4-methylphenyl) acrylamide synthesized in Example 1-3.
4 is a result of NMR identification of (E) -3- (4-fluoro-3-hydroxyphenyl) acrylamide synthesized in Example 1-4.
5 is a result of NMR identification of (E) -4- (3-hydroxy-4-isopropoxyphenyl) but-3-en-2-one synthesized in Example 1-5.
6 is a result of NMR identification of (E) -4- (4-benzyloxy-3-hydroxyphenyl) but-3-en-2-one synthesized in Example 1-6.
7 is a result of NMR identification of (E) -4- (4-allyloxy-3-hydroxyphenyl) but-3-en-2-one synthesized in Example 1-7.
8 is a result of measuring the osteoclast differentiation and proliferation inhibitory ability of compounds including the novel compounds of the present invention KP2 to KP5 and their positive controls Fosamax and Osmundacetone by TRAP assay. (KP2: (E) -4- (3-hydroxy-4-methylphenyl) but-3-en-2-one; KP3: (E) -4- (4-fluoro-3-hydroxyphenyl) butt -3-en-2-one; KP4: (E) -3- (3-hydroxy-4-methylphenyl) acrylamide; KP5: (E) -3- (4-fluoro-3-hydroxyphenyl) Acrylamide)

Hereinafter, the present invention will be described in detail.

However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples.

<Example 1>

Synthesis of novel compounds

In order to identify substances having anticancer activity and osteoclast differentiation inhibitory activity, derivatives were separated and purified from each Benzylideneacetone derivative synthesis mixture, and the chemical structure of each compound was determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS).

The synthesis method of each Benzylideneacetone derivative and the specific NMR and MS analysis results are as follows:

<Example 1-1>

Synthesis of (E) -4- (4-fluoro-3-hydroxyphenyl) but-3-en-2-one (KP3)

Boron tribromide (1M methylene chloride solution. 10 ml) was added dropwise to the methylene chloride solution of 3-methoxy-4-fluorobenzaldehyde (1a) (440 mg, 2.85 mmol) under cooling dropwise. , And stirred at room temperature for 5 hours. The reaction solution was cooled again with ice, and the reaction was stopped by slowly adding cold water, and then a 5N hydrochloric acid solution was added dropwise until the pH reached 1. After the reaction solution was concentrated under reduced pressure, water and ethyl acetate were added to the residue, and the organic layer was separated. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (elution solvent: n-hexane-ethyl acetate 4: 1), and 4-fluoro-3-hydroxybenzaldehyde (2a). 210 mg was obtained.

4-Fluoro-3-hydroxybenzaldehyde (2a, 140.11mg, 1.0mmol) and CuBr ₂ (223.35mg, 1mmol) were dissolved in 5mL acetone at room temperature. The reaction mixture was stirred at 60 ° C. After the mixture was cooled to room temperature, after 6 hours, it was filtered through Celite. The organic layer was concentrated in vacuo, and the product (E) -4- (4-fluoro-3-hydroxyphenyl) but-3-en-2-one (3a, 12%) as a brown solid was used as eluent ethyl acetate and n -Purified by flash chromatography using hexane (1: 4).

The NMR and MS analysis results are as follows (see FIG. 1).

¹ H NMR (400 MHz, CD ₃ OD): δ7.56 (1H, d, J = 16.4 Hz), 7.22 (2H, dd, J = 7.6, 2.1 Hz), 7.13-7.10 (2H, m), 6.67 (1H, J = 16.4 Hz), 2.38 (3H, s); Ms (ESI) m / z : 181.1 [M + H] ⁺ ;

<Example 1-2>

Synthesis of (E) -4- (3-hydroxy-4-methylphenyl) but-3-en-2-one (KP2)

Boron tribromide (1M methylene chloride solution. 5 mL) was added dropwise to a methylene chloride solution of 3-methoxy-4-methylbenzaldehyde (1b) (400 mg, 2.66 mmol) under cooling. , Stirred at room temperature for 5 hours

The reaction solution was cooled again with ice, and the reaction was stopped by slowly adding cold water, and then a 5N hydrochloric acid solution was added dropwise until pH 1 was reached. After the reaction solution was concentrated under reduced pressure, water and ethyl acetate were added to the residue, and the organic layer was separated. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (elution solvent: n-hexane-ethyl acetate 4: 1), and 150 mg of 3-hydroxy-4-methylbenzaldehyde (2b) (41.4%).

3-hydroxy-4-methylbenzaldehyde (2b, 136.1mg, 1.0mmol) and CuBr ₂ (223.35mg, 1mmol) were dissolved in 5mL acetone at room temperature. The reaction mixture was stirred at 60 ° C. After the mixture was cooled to room temperature, after 6 hours, it was filtered through Celite. The organic layer was concentrated in vacuo, and the product (E) -4- (3-hydroxy-4-methylphenyl) but-3-en-2-one (3b, 17%) as a brown solid was used as eluent as ethyl acetate and n-hexane. It was purified by flash chromatography using (1: 4).

The NMR and MS analysis results are as follows (see FIG. 2).

¹ H NMR (700 MHz, CD ₃ OD): δ7.56 (1H, d, J = 16.24 Hz), 7.14 (1H, d, J = 7.42 Hz), 7.03-7.02 (2H, m), 6.67 (1H , J = 16.24 Hz), 2.38 (3H, s), 2.22 (3H, s); Ms (ESI) m / z : 177.1 [M + H] ⁺ ;

<Example 1-3>

Synthesis of (E) -3- (3-hydroxy-4-methylphenyl) acrylamide (KP4)

Step 1: Preparation of ethyl (E) -3- (3-methoxy-4-methylphenyl) acrylate

Sodium hydride (pre-washed with 60% dispersion oil, 300 mg, 7.5 mmol) was stirred in anhydrous THF (10 mL) under nitrogen conditions. To this, triethyl phosphonoacetate (Triethyl phosphonoacetate, 1.345 g, 6 mmol) was added dropwise and stirred for 25 minutes. 3-Methoxy-4-methylbenzaldehyde (3-methoxy-4-methylbenzaldehyde, 1f, 751 mg, 5 mmol) was added dropwise to the THF (3 mL) over about 30 minutes.

The resulting mixed solution was stirred at room temperature for 20 hours, quenched with 100 mL of water, and extracted with ethyl acetate (3 x 100 mL). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated in vacuo. Purification by flash column chromatography (ethyl acetate / n-hexane 1: 4) yielded 2f (700 mg, 63%) of an unsaturated ester in the form of a yellow oil.

Step 2: Preparation of (E) -3- (3-methoxy-4-methylfe) acrylic acid

Ethyl (E) -3- (3-methoxy-4-methylphenyl) acrylate ( 2f, 220mg, 1mmol) dissolved in THF (5mL) was added to sodium hydroxide (2.0M, 5mL) and 2h During heating was refluxed. The mixture was quenched with 10 mL water) and acidified to pH2 with hydrochloric acid (2.0M). Then the solution was extracted with ethyl acetate (3 x 100 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give a white solid ( 3f, 190 mg, 98.8%).

Step 3: Preparation of (E) -3- (3-methoxy-4-methylphenyl) acrylamide

(E) -3- (3-methoxy-4-methylfe) acrylic acid (3f, 100mg, 0.52mmol) and di-tart-butyldicarbonate in dioxane (1mL) of pyridine (0.1 mL) ( (BOC) ₂ O, 227 mg, 1.04 mmol) and ammonium bicarbonate (NH₄ HCO3, 83 mg, 1.04 mmol) were added at once, and the mixture was stirred at 60 ° C for 2 hours.

The reaction mixture was concentrated in vacuo, and the residue was partitioned between ethyl acetate and water. The ethyl acetate phase was washed with 5% aqueous sodium bicarbonate, 0.1N HCl and brine. The organic phase was dried over sodium sulfate, filtered and concentrated in vacuo to give 4f (68mg, 68.3%) as a yellow solid. It was used without further purification.

Step 4: Preparation of (E) -3- (3-hydroxy-4-methylphenyl) acrylamide

Boron tribromide (1M methylene chloride solution. 5 mL) was added to (E) -3- (3-methoxy-4-methylphenyl) acrylamide ((E) -3- (3-methoxy-4-methylphenyl) acrylamide, 4f, 40 mg, 0.21 mmol) was added dropwise to the methylene chloride solution under cooling, followed by stirring at room temperature for 4 hours.

The reaction solution was cooled again with ice, and the reaction was stopped by slowly adding cold water, and then a 5N hydrochloric acid solution was added dropwise until pH 1 was reached. After the reaction solution was concentrated under reduced pressure, water and ethyl acetate were added to the residue, and the organic layer was separated. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (elution solvent: methylene chloride / MeOH 20: 1), and the yellow solid (E) -3- (3-hydroxy-4-methylphenyl) acrylamide ((E) -3- (3-hydroxy-4-methylphenyl) acrylamide, 5f, 18 mg, 48.3%).

The NMR and MS analysis results are as follows (see FIG. 3).

¹ H NMR (400 MHz, MeOD): δ 7.45 (1H, d, J = 15.6 Hz), 7.10 (1H, d, J = 7.6 Hz), 6.98-6.95 (2H, m), 6.53 (1H, J = 15.6 Hz), 2.13 (3H, s); Ms (ESI) m / z : 178.1 [M + H] ⁺ ;

<Example 1-4>

Synthesis of (E) -3- (4-fluoro-3-hydroxyphenyl) acrylamide (KP5)

In the same manner as in Example 1-3, using 3-methoxy-4-fluorobenzaldehyde ( 1 g ) instead of 3-methoxy-4-methylbenzaldehyde in step 1 of Example 1-3 above. This gave the target compound, a yellow solid (E) -3- (4-fluoro-3-hydroxyphenyl) acrylamide ( 5 g , 16 mg, 40%).

Specifically, to give a yellow oil of 2g (680mg, 60.6%) from 1g, and obtaining a 3g (183mg, 93.2%) of a white solid from 2g, and to give a yellow solid 4g (53mg, 53.2%) from 3g From 4 g , a new compound (E) -3- (4-fluoro-3-hydroxyphenyl) acrylamide ( 5 g, 16 mg, 40%) as a yellow solid was obtained.

The NMR and MS analysis results are as follows (see FIG. 4).

¹ H NMR (400 MHz, MeOD): δ 7.45 (1H, d, J = 12.4 Hz), 7.15 (1H, dd, J = 2.0, 2.0 Hz), 7.10-7.01 (2H, m), 6.52 (1H, d, J = 16.0 Hz); Ms (ESI) m / z : 193.1 [M + H] ⁺ ;

<Example 1-5>

Synthesis of (E) -4- (3-hydroxy-4-isopropoxyphenyl) but-3-en-2-one

Step 1: Preparation of 3-hydroxy-4-isopropoxybenzaldehyde

A suspension stirred with 3,4-dihydrobenzaldehyde (250 mg, 1.81 mmol) and anhydrous potassium carbonate (250 mg, 1.81 mmol) in dried dimethylformamide (3 ml) was heated to 70 ° C., and 2-bromopropane. (0.17 ml, 1.81 mmol) was added dropwise under nitrogen conditions for 30 minutes. The mixture was stirred at room temperature overnight, then poured into ice water (50 ml). The mixture solution was extracted with ethyl acetate, and the obtained extract was washed with water and brine, and evaporated in vacuo to obtain a brown oil. The obtained residue was purified by silica gel column chromatography (ethyl acetate / n-hexane 1: 4) to give the product 2h (46mg, 50%) as a brown solid.

Step 2: Preparation of (E) -4- (3-hydroxy-4-isopropoxyphenyl) but-3-en-2-one

3-hydroxy-4-isopropoxybenzaldehyde ( 2h, 50mg, 0.277mmol) and CuBr2 (62mg, 0.277mmol) were dissolved in 3mL of acetone at room temperature. The reaction mixture was stirred at 60 ° C. After 6 hours, the mixture was cooled to room temperature and filtered through Celite. The organic layer was concentrated in vacuo, and the eluent was purified by flash chromatography using ethyl acetate and n-hexane (1: 4). As a result, a new compound (E) -4- (3-hydroxy-4-isopropoxyphenyl) but-3-en-2-one ( 3h , 33mg, 53.3%) as a white solid was obtained.

Of the product 3h The NMR and MS analysis results are as follows (see FIG. 5).

¹ H NMR (400 MHz, CDCl ₃ ) δ7.44 (1H, d, J = 16.0 Hz), 7.17 (1H, d, J = 2.4 Hz), 7.05 (1H, dd, J = 8.4 Hz, 6.0 Hz) , 6.87 (1H, d, J = 8.4 Hz), 6.60 (1H, d, J = 16.0 Hz), 5.74 (1H, s), 4.67 (1H, hept, J = 6.4 Hz) 2.37 (3H, s), 1.41 (6H, doublet of doublets, J = 6.0 Hz)); Ms (ESI) m / z : 221.1 [M + H] ⁺ ;

<Example 1-6>

Synthesis of (E) -4- (4- (benzyloxy) -3-hydroxyphenyl) but-3-en-2-one

In step 1 of Example 1-5, using benzyl bromide instead of 2-bromopropane, the target compound, 4- (benzyloxy) -3-hydroxybenzaldehyde, was prepared in the same manner as in Example 1-5. (2i) was obtained.

4- (benzyloxy) -3-hydroxybenzaldehyde (2i, 50mg, 0.277mmol) and CuBr2 (48.9mg, 0.22mmol) were dissolved in 3mL of acetone at room temperature. The reaction mixture was stirred at 60 ° C. After 6 hours, the mixture was cooled to room temperature and filtered through Celite. The organic layer was concentrated in vacuo, and the eluent was purified by flash chromatography using ethyl acetate and n-hexane (1: 4). As a result, a new compound (E) -4- (4- (benzyloxy) -3-hydroxyphenyl) but-3-en-2-one (3i, 37mg, 62.6%) as a white solid was obtained.

The NMR and MS analysis results are as follows (see FIG. 6).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.46 7.42 (6H, m), 7.19 (1H, d, J = 2.0 Hz), 7.05 (1H, dd, J = 8.4 Hz, 2.4 Hz), 6.95 (1H, d, J = 8.4 Hz), 6.95 (1H, d, J = 8.4 Hz), 6.61 (1H, d, J = 16.0 Hz), 5.71 (1H, s), 5.17 (2H, s), 2.37 (3H, s); Ms (ESI) m / z : 269.2 [M + H] ⁺ ;

<Example 1-7>

Synthesis of (E) -4- (4- (allyloxy) -3-hydroxyphenyl) but-3-en-2-one

In step 1 of Example 1-5, using bromopropene instead of 2-bromopropane, 4- (allyloxy) -3-hydroxyben, the target compound, was prepared in the same manner as in Example 1-5. Zaldehyde (2j) was obtained.

4- (allyloxy) -3-hydroxybenzaldehyde (2j, 50mg, 0.28mmol) and CuBr2 (62.7mg, 0.28mmol) were dissolved in 3mL of acetone at room temperature. The reaction mixture was stirred at 60 ° C. After 6 hours, the mixture was cooled to room temperature and filtered through Celite. The organic layer was concentrated in vacuo, and the eluent was purified by flash chromatography using ethyl acetate and n-hexane (1: 4). As a result, a new compound (E) -4- (4- (allyloxy) -3-hydroxyphenyl) but-3-en-2-one (3j, 34 mg, 55.6%) as a white solid was obtained.

The NMR and MS analysis results are as follows (see FIG. 7).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.43 (1H, d, J = 16.4 Hz), 7.18 (1H, d, J = 2.4 Hz), 7.04 (1H, dd, J = 8.4 Hz, 2.0 Hz), 6.87 (1H, d, J = 8.0 Hz), 6.60 (1H, d, J = 16.0 Hz), 6.11 6.04 (1H, m), 5.80 (1H, s), 5.43 (1H, dd, J = 17.2 Hz, 12.8 Hz), 5.36 (1H, dd, J = 10.4 Hz, 8.0 Hz), 4.67 4.65 (2H, m), 2.37 (3H, s); Ms (ESI) m / z : 219.2 [M + H] ⁺ ;

<Example 2>

MTT assay

Cytotoxicity to normal cells and cancer cells was confirmed by using the MTT assay for the novel Benzylideneacetone derivatives KP2, KP3, KP4, and KP5 prepared in <Example 1>.

The MTT assay method is as follows.

Normal cells (RAW264.7 mouse macrophage cell line (osteoblast progenitor cell) and NIH3T3 mouse fibroblast line) and cancer cells (AGS human gastric cancer cell line, A549 human lung cancer cell line, HepG2 human liver cancer cell line, HCT116 human colon cancer cell line, PC3 human prostate cancer) Cell line, Caki-1 human kidney cancer cell line, T24 human bladder cancer cell line, HT1080 human fibrosarcoma cell line, B16F10 murine melanoma cell line) 5% CO in 96-well plate containing DMEM with 10% fetal bovine serum (FBS) After incubation at ₂ , 37 ° C, Benzylideneacetone derivatives and controls (osmundacetone and 4-hydroxybenzalacetone) were added to the cell medium and incubated for 48 hours. Then, 100 μL of MTT (0.5 mg / mL phosphate buffered saline) was administered, followed by incubation for 2 hours. After adding 100 μL of DMSO to each well and incubating for 10 minutes, absorbance was measured at 550 nm using a microplate reader (SPCTRA MAX 340PC, Molecular Devices, USA). Absorbance is an index indicating the number of surviving cells, calculated by the formula below, and confirmed reproducibility in three experiments.

Cell proliferation rate (%) = OD550 (sample) / OD550 (control)

Results of normal cell MTT assay of Benzylideneacetone derivatives Compound LD ₅₀ (μ) Normal cell RAW264.7 NIH3T3 Osmundacetone 529 ± 41 > 5000 4-Hydroxybenzalacetone 294 ± 45 201 ± 10 KP2 395 ± 22 688 ± 12 KP3 503 ± 57 > 5000 KP4 > 5000 > 5000 KP5 > 5000 > 5000

As can be seen in Table 9, Benzylideneacetone derivative KP2 to LD ₅₀ for the normal cells of the derivative calculation Benzylideneacetone KP5 after administration on the basis of changes in cell jeungsikyul was similar to that of LD ₅₀ value of Osmundacetone. KP2 only showed weak cytotoxicity. In the case of the macrophage cell line RAW264.7, KP2 and KP3 show weak cytotoxicity, so these compounds are thought to have weak immunosuppressive functions. Therefore, it can be seen that the derivatives of the present invention having low cytotoxicity to normal cells can be safely used as pharmaceutical and food compositions.

Cancer cell MTT assay results of Benzylideneacetone derivatives Com
pound LD ₅₀ (μ) Cancer cell AGS A549 HepG2 HCT116 PC3 Caki-1 T24 HT1080 B16F10 Osmund
acetone 56.1 ± 2.1 > 5000 > 5000 689 ± 27 60.6 ± 14 695 ± 14 2000 ± 250 > 5000 64.9 ± 59 KP2 209 ± 5.9 471 ± 24 339 ± 79 380 ± 32 309 ± 3.1 387 ± 57 1120 ± 67 256 ± 14 298 ± 27 KP3 239 ± 14 1090 ± 46 612 ± 95 389 ± 18 387 ± 48 408 ± 12 > 5,000 379 ± 10 278 ± 15 KP4 329 ± 14 989 ± 50 379 ± 38 315 ± 19 394 ± 140 361 ± 16 1040 ± 360 309 ± 10 1210 ± 110 KP5 569 ± 4.8 935 ± 10 1280 ± 390 > 5,000 > 5,000 2990 ± 58 203 ± 69 1090 ± 19 > 5,000

As shown in Table 9, Benzylideneacetone derivative KP2 to KP5 administered after LD on of the Benzylideneacetone derivative calculated on the basis of a change in cell jeungsikyul cancer cells ₅₀ exhibited a stronger anti-cancer activity in some cancer compared to the LD ₅₀ value of Osmundacetone.

Specifically, for A549 lung cancer cells, KP2 showed significantly superior cancer cell inhibitory activity than Osmundacetone, and in HepG2 liver cancer cells, KP2 and KP4 showed particularly excellent cancer cell inhibitory activity, and in HCT116 colorectal cancer cells, KP2 to KP4 except KP5 It was confirmed that the inhibitory activity was more than twice that of Osmundacetone. In Caki-1 kidney cancer cells, all of the derivatives except KP5 were found to have superior inhibitory activity than Osmundacetone, and it was confirmed that K24 was particularly superior to T24 bladder cancer cells. Finally, for HT1080 fibrosarcoma cancer cells, it was confirmed that all of the derivatives except KP5 had better inhibitory activity than the positive control Osmundacetone.

Therefore, it was confirmed that the inhibitory activity on human lung cancer, colorectal cancer, kidney cancer, bladder cancer, and fibrosarcoma cancer cells was superior to or similar to that of Osmundacetone. .

<Example 3>

Inhibition activity of osteoclast proliferation and differentiation

Benzylideneacetone derivatives prepared in <Example 1> confirmed the activity of proliferation and differentiation inhibition on osteoclasts using a TRAP assay (tartrate-resistant acid phosphatase), a specific osteoclast staining method.

The specific method of the TRAP assay is as follows.

1. Bone marrow cell culture

The tibia and femur of male C57BL / 6 mice aged 6-8 weeks were aseptically excised, and bone marrow cells were collected aseptically with a syringe (21G, Korea Green Cross). Bone marrow cells were suspended in 500 μ of α-MEM medium (Gibco BRL Co.) containing sodium bicarbonate (2.0 g / L), streptomycin (100 mg / L), and penicillin (100,000 unit / mL) and dispensed into 48 well plates. The assay was performed with triplicate. The monocytes, which are progenitor cells of osteoclasts, were separated and treated with RANKL and M-CSF, which are differentiation promoting factors, to differentiate into osteoclasts within 5-7 days.

2. Measurement of osteoclast differentiation

1) Sample preparation: KP2 to KP5 Benzylideneacetone derivatives (0.5μ1μ5μ or 10μFosamax (0.5μ1μ5μ or 10μ, etc.) were dissolved in dimethylsulfoxide (DMSO) or sterile distilled water at appropriate concentrations, respectively. Did.

2) Sample administration: The sample was continuously administered to the medium with 1:20 (v / v; sample 25 μ per 500 μ medium) from the first day of bone marrow cell culture, and the medium was replaced every 3 days.

3) Measurement of osteoclast differentiation: Osteoblasts were defined as TRAP-positive multinucleated cells stained with TRAP. The TRAP staining solution was dissolved in 5 mg of naphthol AS-MS phosphate (Sigma N-4875) and 25 mg of Fast Red Violet LB salt, a color development reagent, in about 0.5 mL of N, N-dimethylformamide, and 0.1 containing 50 mM tartaric acid. It was mixed with N NaHC0 ₃ buffer solution (50 mL). Reaction reagents were stored in the refrigerator until use.

Bone marrow cells were cultured for 6-7 days in a medium containing a differentiation promoting factor, and then the medium was removed, washed with PBS, and fixed with PBS containing 10% formalin for 2-5 minutes. After that, it was fixed for 1 minute with a 1: 1 mixed solution of ethanol and acetone, and dried. The immobilized cells were treated with TRAP staining solution for 6 minutes at 37 ^o C while blocking light, washed with PBS, and then hematoxylin staining was performed, and the degree of staining of the cells was observed under a microscope.

Of the TRAP-positive cells in the microscope field, cells having two or more nuclei were determined as osteoclasts and the number of cells was measured. Osteoclast differentiation inhibiting effect of Benzylideneacetone derivative was calculated from the 50% inhibitory concentration IC ₅₀ as compared to the control group.

As can be seen in Figure 8, in the group treated with Benzylideneacetone derivatives, the formation of giant osteoclasts, which are multinucleated cells, was significantly suppressed similarly to the positive control group (group to which Osmundacetone or Fosamax was added to the culture medium), and in the case of KP2, Osmundacetone It showed an IC ₅₀ value that is about 3 times better than the group to which was added. In addition, proliferation of osteoclast progenitor cells was also significantly suppressed, and it was observed that proliferation inhibitory effect as well as osteoclast differentiation was very large.

Since the novel Benzylideneacetone derivatives according to the present invention show strong proliferation and differentiation inhibitory activity against osteoclasts that cause bone loss in addition to cancer cell specific cytotoxicity, safe and effective anticancer drugs and osteoporosis prevent and treat or improve bone diseases It can be usefully used to develop food products.

Claims (15)

delete (E) -4- (4-fluoro-3-hydroxyphenyl) but-3-en-2-one, (E) -4- (3-hydroxy-4-methylphenyl) but-3-ene- Benzylidene selected from the group consisting of 2-one, (E) -3- (3-hydroxy-4-methylphenyl) acrylamide and (E) -3- (4-fluoro-3-hydroxyphenyl) acrylamide. Acetone derivatives or pharmaceutically acceptable salts thereof.
delete (E) -4- (4-fluoro-3-hydroxyphenyl) but-3-en-2-one, (E) -4- (3-hydroxy-4-methylphenyl) but-3-ene- One or more selected from the group consisting of 2-one, (E) -3- (3-hydroxy-4-methylphenyl) acrylamide and (E) -3- (4-fluoro-3-hydroxyphenyl) acrylamide A pharmaceutical composition for preventing or treating cancer, comprising a benzylidene acetone derivative or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 4, wherein the cancer is breast cancer, colon cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, melanoma in the skin or eye, uterine cancer, ovarian cancer, rectal cancer, Anorectal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penis Selected from the group consisting of cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or urinary tract cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma and pituitary adenoma A pharmaceutical composition for preventing or treating cancer, characterized in that it is one or more diseases.
A pharmaceutical composition for preventing or treating bone disease comprising a benzylidene acetone derivative represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient;
<Formula 1>

In the above formula, R1 and R2 are different,
R1 and R2 are each independently -H, -OH, -SH, C _1-4 linear or branched alkyl group, C _1-4 linear or branched alkoxy group, halogen, allyloxy group, allyloxy, benzyloxy group ( benzyloxy), an aryloxy group having one or more selected from the group consisting of heteroatoms and substituents, and a heterocycloalkyl group having 3-7 atoms having one or more heteroatoms, wherein the heteroatom is Is composed of O, N and S:
R3 is a C _1-4 straight-chain or branched alkyl group, -NHR4, -N (R4) ₂ , OH, respectively:
R4 is each independently hydrogen or a C _1-4 straight or branched alkyl group.
The benzylidene acetone derivative represented by Chemical Formula 1 is (E) -4- (4-fluoro-3-hydroxyphenyl) but-3-en-2-one, (E) -4. -(3-hydroxy-4-methylphenyl) but-3-en-2-one, (E) -3- (3-hydroxy-4-methylphenyl) acrylamide, (E) -3- (4-fluoro Low-3-hydroxyphenyl) acrylamide, (E) -4- (4-mercapto-3-hydroxyphenyl) but-3-en-2-one, (E) -4- (4-hydroxy -3-mercaptophenyl) but-3-en-2-one, (E) -4- (3-hydroxy-4-isopropoxyphenyl) but-3-en-2-one, (E)- 4- (4-benzyloxy-3-hydroxyphenyl) but-3-en-2-one, (E) -4- (4-allyloxy-3-hydroxyphenyl) but-3-ene-2- On, (E) -4- (3-hydroxy-4- (4-methylpiperazin-1-yl) phenyl) but-3-den-2-one, (E) -4- (3-hydroxy -4- (pyrrolidin-1-yl) phenyl) but-3-en-2-one, (E) -4- (3-hydroxy-4- (4-hydroxyphenoxy) phenyl) but- 3-ene-2-one, selected from the group consisting of (E) -4- (3-hydroxy-4- (piperidin-4-yloxy) phenyl) but-3-en-2-one One or more benzylidene bone disease prophylactic or therapeutic pharmaceutical composition, characterized in that acetone derivative.
The method of claim 6, wherein the bone disease is osteoporosis, Paget's disease, rickets, osteomalacia, nephrotic dystrophy in patients with renal failure, joint pain, fractures, rheumatoid bone disease, degenerative bone disease, periodontal disease, inflammatory alveolar bone absorption disease and inflammatory bone. A pharmaceutical composition for preventing or treating bone disease, which is selected from the group consisting of absorbing diseases.
Food composition for preventing or improving bone disease comprising a benzylidene acetone derivative represented by the following formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient;
<Formula 1>

In the above formula, R1 and R2 are different,
R1 and R2 are each independently -H, -OH, -SH, C _1-4 linear or branched alkyl group, C _1-4 linear or branched alkoxy group, halogen, allyloxy group, allyloxy, benzyloxy group ( benzyloxy), an aryloxy group having one or more selected from the group consisting of heteroatoms and substituents, and a heterocycloalkyl group having 3-7 atoms having one or more heteroatoms, wherein the heteroatom is Is composed of O, N and S:
R3 is a C _1-4 straight-chain or branched alkyl group, -NHR4, -N (R4) ₂ , OH, respectively:
R4 is each independently hydrogen or a C _1-4 straight or branched alkyl group.
10. The method of claim 9, wherein the bone disease is osteoporosis, Paget's disease, rickets, osteomalacia, nephrotic dystrophy of renal failure patients, joint pain, fracture, rheumatoid bone disease, degenerative bone disease, periodontal disease, inflammatory alveolar bone resorption disease and inflammatory bone Food composition for preventing or improving bone disease, characterized in that selected from the group consisting of absorption diseases.
(E) -4- (4-fluoro-3-hydroxyphenyl) but-3-en-2-one, (E) -4- (3-hydroxy-4-methylphenyl) but-3-ene- One or more selected from the group consisting of 2-one, (E) -3- (3-hydroxy-4-methylphenyl) acrylamide and (E) -3- (4-fluoro-3-hydroxyphenyl) acrylamide Food composition for preventing or improving cancer, comprising a benzylidene acetone derivative or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 11, wherein the cancer is breast cancer, colon cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, Anorectal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penis Selected from the group consisting of cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or urinary tract cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma and pituitary adenoma Food composition for preventing or improving cancer, characterized in that it is at least one disease.
As shown in Method 1 below,
Hydrolyzing the methyl ether group of compound 1 under acid catalyst conditions to prepare compound 2 (step a); And
Method for producing a benzylidene acetone derivative of the following compound 3 comprising the step of preparing a compound 3 by reacting the compound ₂ prepared in step a with CuBr ₂ (step b);
[Method 1]

R of method 1 is -H, -OH, -SH, C _1-4 straight or branched alkyl group, C _1-4 straight or branched alkoxy group, halogen, allyloxy group (allyloxy), benzyloxy group (benzyloxy ), Aryloxy having one or more selected from the group consisting of heteroatoms and substituents, and one selected from the group consisting of 3-7 membered heterocycloalkyl groups having one or more heteroatoms, wherein the heteroatom is O , N and S.
As shown in Method 2 below,
Preparing compound 2 'by reacting the following compound 1' with triethyl phosphonoacetate (step a ');
Preparing a compound 3 'by reacting the ethyl ester group of compound 2' with a base (step b ');
Preparing a compound 4 'by reacting a hydroxy group of compound 3' with NH ₄ HCO ₃ (step c '); And
Method for producing a benzylidene acetone derivative of the following compound 5 'comprising the step (step d') of hydrolysis of the methyl ether group of the following compound 4 'under acid catalyst conditions (step d');
[Method 2]

R of the method 2 is -H, -OH, -SH, C _1-4 straight or branched chain alkyl group, C _1-4 straight or branched alkoxy group, halogen, allyloxy group (allyloxy), benzyloxy group (benzyloxy ), Aryloxy having one or more selected from the group consisting of heteroatoms and substituents, and one selected from the group consisting of 3-7 membered heterocycloalkyl groups having one or more heteroatoms, wherein the heteroatom is O , N and S.
As shown in Method 3 below,
Preparing compound 2 "in which a protecting group is selectively introduced by using a halogenated alkyl compound under the basic conditions of compound 1" (step a "); and
Method for producing a benzylidene acetone derivative of the following compound 3 "comprising the step (step b") of reacting compound 2 "prepared in step a" with CuBr ₂ to prepare compound 3 ";
[Method 3]

R of the method 3 is C _1-4 straight or branched chain alkyl group, C _1-4 straight or branched alkoxy group, C _1-4 alkenyl group; Aryl group having one or more selected from the group consisting of heteroatoms and substituents and 3-7 membered heterocycloalkyl groups having one or more heteroatoms, wherein the heteroatoms are O, N and S Is done.

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