WO2017018750A1 - Novel compound having blt inhibitory activity and composition, for preventing or treating inflammatory diseases, comprising same as active ingredient - Google Patents

Novel compound having blt inhibitory activity and composition, for preventing or treating inflammatory diseases, comprising same as active ingredient Download PDF

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Publication number
WO2017018750A1
WO2017018750A1 PCT/KR2016/008069 KR2016008069W WO2017018750A1 WO 2017018750 A1 WO2017018750 A1 WO 2017018750A1 KR 2016008069 W KR2016008069 W KR 2016008069W WO 2017018750 A1 WO2017018750 A1 WO 2017018750A1
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Prior art keywords
prop
ynyl
phenyl
carbonyl
fluorophenyl
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PCT/KR2016/008069
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French (fr)
Korean (ko)
Inventor
최용석
김재홍
이경
한효경
위준동
권진선
구자일
Original Assignee
동국대학교 산학협력단
고려대학교 산학협력단
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Priority claimed from KR1020160093762A external-priority patent/KR101796391B1/en
Application filed by 동국대학교 산학협력단, 고려대학교 산학협력단 filed Critical 동국대학교 산학협력단
Priority to JP2018503553A priority Critical patent/JP6574517B2/en
Priority to ES16830783T priority patent/ES2834549T3/en
Priority to EP16830783.3A priority patent/EP3327000B1/en
Priority to CN201680055514.4A priority patent/CN108349874B/en
Priority to US15/745,337 priority patent/US10179764B2/en
Publication of WO2017018750A1 publication Critical patent/WO2017018750A1/en
Priority to US16/181,953 priority patent/US10766857B2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4965—Non-condensed pyrazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/02—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having nitrogen atoms of carboxamide groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals
    • C07C233/04—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having nitrogen atoms of carboxamide groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals with carbon atoms of carboxamide groups bound to acyclic carbon atoms of an acyclic saturated carbon skeleton
    • C07C233/07—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having nitrogen atoms of carboxamide groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals with carbon atoms of carboxamide groups bound to acyclic carbon atoms of an acyclic saturated carbon skeleton having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a six-membered aromatic ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
    • C07D241/04—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having no double bonds between ring members or between ring members and non-ring members
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D265/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one oxygen atom as the only ring hetero atoms
    • C07D265/28—1,4-Oxazines; Hydrogenated 1,4-oxazines
    • C07D265/30—1,4-Oxazines; Hydrogenated 1,4-oxazines not condensed with other rings

Definitions

  • the present invention relates to a novel compound and its use, and more particularly, to a novel compound exhibiting inhibitory activity of Leukotriene B4 receptor 2 (BLT2) and a pharmaceutical composition for preventing or treating inflammatory diseases comprising the same as an active ingredient.
  • BLT2 Leukotriene B4 receptor 2
  • Inflammatory reactions are one of the body's immune systems that are activated through various mechanisms of action to protect against physical or chemical agents, bacterial infections, and immunological stimuli. However, if such an inflammatory response persists, it promotes mucosal damage, thereby causing inflammatory diseases such as rheumatoid arthritis, arteriosclerosis, gastritis, and asthma due to redness, fever, swelling, pain, and dysfunction. have. These inflammatory reactions are classified into acute and chronic inflammations over time, and acute inflammations can cause symptoms such as erythema, fever, pain, and edema, while inflammatory reactions last several days to several weeks. Is a prolonged inflammatory state, sometimes over several years to decades, and is accompanied by histological changes such as invasion of monocytes, proliferation of fibroblasts and capillaries, fibrosis due to increased connective tissue, and tissue destruction. .
  • pro-inflammatory cytokines pro-inflammatory cytokines
  • interferon-gamma INF- ⁇
  • tumor necrosis factor- ⁇ TNF- ⁇
  • interleukin-1 interleukin-1, IL-1
  • interleukin during inflammatory processes in the body INF- ⁇
  • cytokines such as -6 (interleukin-6, IL-6), nitric oxide (NO) and prostaglandin E2 (PGE2)
  • cytokines such as -6 (interleukin-6, IL-6), nitric oxide (NO) and prostaglandin E2 (PGE2) are well known as major inflammatory agents.
  • leukotriene B4 is a group of inflammatory lipid mediators synthesized from arachidonic acid (AA) by the 5-lipoxygenase pathway that mediates acute and chronic inflammation.
  • LTB 4 is known to have a biological effect by binding to two types of receptors, BLT1 and BLT2.
  • Leukotriene B4 receptor 2 (BLT2) is a group of G protein-coupled receptors (GPCRs) that has low affinity for LTB 4 and is a lipid mediator of arachidonic acid (AA) induced through a 5-lipoxygenase dependent pathway. to be.
  • the present inventors while continuing to research to develop a substance that induces more effective termination of inflammation, in order to solve the conventional problems as described above, has produced a novel compound showing a BLT2 inhibitory activity, including the compound It was the first to develop an inflammatory disease treatment.
  • the present invention has been made to solve the above problems, the present inventors have confirmed the therapeutic effect of the inflammatory disease of the novel compound showing the BLT2 inhibitory activity and completed the present invention based on this.
  • Another object of the present invention is to provide a pharmaceutical composition for preventing or treating inflammatory diseases, comprising the novel compound or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the present invention provides a novel compound or a pharmaceutically acceptable salt thereof exhibiting BLT2 inhibitory activity.
  • the compound is tert -butyl 4- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate; N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide; N- (3- (4-methylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N- (3- (4- (4-ethylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N- (3- (4- (4-isopropylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N- (3- (4- (4- (2-hydroxy
  • the present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease, comprising the novel compound or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the inflammatory disease may be selected from the group consisting of asthma, atherosclerosis, cancer, itching of the skin, rheumatoid arthritis and inflammatory bowel disease.
  • the composition may inhibit Leukotriene B4 receptor 2 (BLT2) activity.
  • BLT2 Leukotriene B4 receptor 2
  • It provides a method of treating an inflammatory disease comprising administering the pharmaceutical composition to a subject.
  • the present invention provides a therapeutic use of an inflammatory disease of a composition comprising the novel compound or a pharmaceutically acceptable salt thereof.
  • the present invention relates to a novel compound that exhibits Leukotriene B4 receptor 2 (BLT2) inhibitory activity and a pharmaceutical composition for preventing or treating an inflammatory disease comprising the same.
  • BLT2 Leukotriene B4 receptor 2
  • the present inventors have identified a novel compound that exhibits BTL2 inhibitory activity in order to solve in vivo instability and difficulty in mass production, which are problems of conventional inflammatory disease treatment substances, and has excellent cancer cell death enhancement and metastasis suppression effect and chemotaxis inhibition of the compound.
  • the effects, and anti-asthma effects have been confirmed experimentally, it is expected that it can be usefully used as a pharmaceutical composition for treating inflammatory diseases.
  • 1A to 1E show the results of confirming the growth inhibitory effect of the compound of the present invention in cells expressing BLT2 (CHO-BLT2).
  • Figures 2a and 2b is a result of confirming the chemotaxis inhibitory effect and IC 50 (50% inhibitory concentration) of the cells by the compound treatment of the present invention in CLT-expressing cells (CHO-BLT2 cells).
  • 3A and 3B show the results of confirming the chemotaxis inhibitory effect of cells treated with the compound of the present invention in cells expressing BLT2 (CHO-BLT2 cells) or cells expressing BLT1 (CHO-BLT1).
  • Figures 4a and 4b is a result of confirming the effect of inhibiting the binding of LTB 4 and BLT2 by the compound treatment of the present invention in BLT2-expressing cells (CHO-BLT2 cells).
  • Figures 5a and 5b is the result of confirming the inhibitory effect of the production of active oxygen by the compound treatment of the present invention in MDA-MB-231 cells or MDA-MB-435 cells.
  • Figures 6a and 6b is a result of confirming the inhibitory effect of IL-8 (interleukin-8) expression by the compound treatment of the present invention in MDA-MB-231 cells or MDA-MB-435 cells.
  • Figures 7a and 7b is a result of confirming the cancer cell infiltration inhibition effect by the compound treatment of the present invention in MDA-MB-231 cells or MDA-MB-435 cells.
  • 13A to 13C show that asthma-induced mice reduced total cells and neutrophils introduced into the abdominal cavity of the mouse by the compound treatment of the present invention.
  • 14A and 14B are results confirming that the total cells and neutrophils introduced into the abdominal cavity of the mouse by the compound treatment of the present invention are reduced in asthma-induced mice.
  • the present inventors when treated with the novel compound prepared in Example, can significantly inhibit the growth of BLT2 expressing cells, thereby promoting cancer cell death, inhibiting cancer cell metastasis, inhibiting BLT2-dependent chemotaxis and The asthma effect, etc. were specifically confirmed and the present invention was completed based on this.
  • the present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.
  • R 1 is C 1 -C 10 is alkyl
  • R 2 is , , , or Is
  • R a is , , , , , , , , , , , , , , , , , , , , , , , , , , , , Or hydroxy,
  • R b is , , or ego
  • R C is , or ego
  • R d is hydrogen or ego
  • R e is or ego
  • R 3 may be hydrogen or fluorine.
  • Preferred examples of the compound represented by Formula 1 according to the present invention are as follows:
  • 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.
  • 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.
  • Acid addition salts according to the invention are dissolved in conventional methods, for example, the compounds represented by the formulas (1) to (4) in an excess of aqueous acid solution, and the salts are water miscible organic solvents 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.
  • water miscible organic solvents 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).
  • a novel compound exhibiting BLT2 inhibitory activity was prepared a novel compound exhibiting BLT2 inhibitory activity (see Examples 1 to 46), and confirmed the growth inhibition of BLT2 expressing cells by the novel compound treatment (see Experimental Example 2). In addition, it was confirmed that chemotaxis of BLT2 expressing cells can be suppressed (see Experimental Example 3).
  • the compound was used to confirm the inhibitory effect of LTB 4 and BLT2 binding (see Experimental Example 4), inhibited intracellular free radicals, inhibited IL-8 expression, inhibited cancer cell infiltration and inhibited metastasis of cancer cells (Experimental Example). 5), the effect of reducing airway hypersensitivity, inhibiting IL-4 production, and inhibiting immune cell influx into the mouse abdominal cavity was specifically confirmed in asthma-induced mice (see Experimental Example 6). It was confirmed that it can be used.
  • the present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease, comprising the compound or a pharmaceutically acceptable salt thereof as an active ingredient.
  • prevention means any action that inhibits or delays the development of an inflammatory disease by administration of a pharmaceutical composition according to the present invention.
  • treatment means any action that improves or advantageously alters the symptoms of an inflammatory disease by administration of a pharmaceutical composition according to the present invention.
  • the inflammatory disease is due to overexpression of Leukotriene B4 receptor 2 (BLT2), and may be at least one selected from asthma, atherosclerosis, cancer, skin itch, rheumatoid arthritis and inflammatory bowel disease, but is limited thereto. It is not.
  • BLT2 Leukotriene B4 receptor 2
  • all BLT2-associated inflammatory diseases known in the art are considered to be included in inflammatory diseases that can be prevented or treated with a compound having the structure of Formula 1 of the present invention.
  • the cancer may be any cancer caused by overexpression of BLT2 or the oncogene Ras.
  • the cancer may be selected from the group consisting of bladder cancer, prostate cancer, pancreatic cancer, breast cancer, brain tumor, skin cancer, and liver cancer, but is not limited thereto.
  • BLT2 Leukotriene B4 receptor 2
  • LTB 4 is one of the group GPCR (G protein-coupled receptor)
  • GPCR G protein-coupled receptor
  • the composition of the present invention can prevent or treat inflammatory diseases by inhibiting cell growth by BLT2. More specifically, the inhibition of the production of ROS induced by BLT2 activity may inhibit LTB 4 -induced chemotaxis.
  • the term “inhibition” means inhibiting any step of transcription, mRNA processing, translation, translocation, and maturation of a gene, or inhibition of protein-to-protein binding, activation of a protein, or signaling through it. .
  • the pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient.
  • 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.
  • it may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like.
  • 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.
  • 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 single or multiple administrations. 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.
  • 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.
  • 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.
  • the present invention also provides a method of treating an inflammatory disease comprising administering the pharmaceutical composition to a subject.
  • subject 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. .
  • Step 1 N -Phenylpentaneamide ( N -phenylpentanamide)
  • Step 2 tert -Butyl 4- (4- Bromobenzoyl Piperazine-1- Carboxylate ( tert -butyl Preparation of 4- (4-bromobenzoyl) piperazine-1-carboxylate)
  • Isopropylethylamine (DIPEA) (2.34 ml, 13.44 mmol) was added and stirred at room temperature for 15 hours.
  • the reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure.
  • Step 3 tert -Butyl 4- (4- (3- Hydroxyprop -One- Inil ) Benzoyl Piperazine-1- Carboxylate ( tert -butyl 4- (4- (3- hydroxyprop -One- ynyl ) benzoyl ) piperazine -1-carboxylate)
  • Triethyl tert -butyl 4- (4-bromobenzoyl) piperazine-1-carboxylate (1.00 g, 2.71 mmol) and propagyl alcohol (0.32 ml, 5.42 mmol) obtained in step 2 were obtained. Dissolved in amine (12 ml) and stirred for 5 minutes. Bis (triphenylphosphine) palladium (II) dichloride (190 mg, 0.271 mmol) and copper iodide (I) (52 mg, 0.271 mmol) were added to the mixture, which was heated and refluxed at 60 ° C. for 17 hours. .
  • the reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure.
  • Step 4 tert -Butyl 4- (4- (3- ( Methylsulfonyloxy ) Prof -One- Inil ) Benzoyl Piperazine-1-carboxylate ( tert -butyl 4- (4- (3- ( methylsulfonyloxy ) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
  • Step 5 tert - Butyl 4 -(4- (3- ( N - Phenylpentaneamido ) Prof -One- Inil ) Benzoyl Piperazine-1-carboxylate ( tert -butyl 4- (4- (3- ( N - phenylpentanamido ) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
  • the reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure.
  • Piperazine-1-carboxylate ( tert- butyl 4- (4- (3- ( N- phenylpentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (382 mg, 73% yield).
  • N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (33.3 mg, 0.072 mmol) obtained in Example 2 and potassium hydroxide ( KOH) (9.09 mg, 0.108 mmol) was dissolved in N , N -dimethylformamide (DMF) (1 ml) and stirred at room temperature for 5 minutes. Iodomethane (9 ⁇ l, 0.144 mmol) was added to the mixture, followed by stirring at the same temperature for 17 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine.
  • KOH potassium hydroxide
  • N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (24.8 mg, 0.061 mmol) obtained in Example 3 and potassium hydroxide ( 8.62 mg, 0.154 mmol) was dissolved in N , N -dimethylformamide (DMF) (1 ml) and stirred at room temperature for 5 minutes. Iodoethane (20 ⁇ l, 0.246 mmol) was added to the mixture and stirred for 17 hours at the same temperature. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine.
  • DMF N -dimethylformamide
  • N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (106 mg, 0.263 mmol) and sodium hydrogencarbonate obtained in Example 2
  • N , N -dimethylformamide (DMF) (2 ml) was added thereto and stirred for 1 hour.
  • 2-iodopropane (30 ⁇ l, 0.316 mmol) was added to the mixture, ice was removed, and the mixture was heated and refluxed at 60 ° C. for 24 hours.
  • N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (56 mg, 0.139 mmol) obtained in Example 2 and potassium carbonate ( 77 mg, 0.556 mmol) was dissolved in acetonitrile (3 ml) and stirred at room temperature for 5 minutes. 2-bromoethanol (99 ⁇ l, 1.39 mmol) was added to the mixture, and the mixture was heated and refluxed at 60 ° C. for 17 hours. The reaction solution was cooled to room temperature, filtered to remove solids, and then concentrated under reduced pressure.
  • N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) obtained in Example 2 and potassium carbonate ( 51 mg, 0.372 mmol) was dissolved in N , N -dimethylformamide (DMF) (2 ml) and then stirred at room temperature for 5 minutes.
  • Cyclopropylmethyl bromide (15 ⁇ l, 0.145 mmol) was added to the mixture, heated at 80 ° C., refluxed and stirred for 4 hours. The reaction solution was cooled to room temperature, filtered to remove solids, and then concentrated under reduced pressure.
  • N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) obtained in Example 2 and iodocyclo
  • N - (3- (4- ( 4-cyclohexylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) - N -phenylpentanamide was obtained (20 mg, 33% yield ).
  • N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) and bromomethyl obtained in Example 2 above Using cyclohexane (20 ⁇ l, 0.145 mmol) in the same manner as in Example 7, the final product, N- (3- (4- (4- (cyclohexylmethyl) piperazin-1-carbonyl) phenyl) prop profile-2-ynyl) - N - phenyl pentane amide (N - (3- (4- ( 4- (cyclohexylmethyl) piperazine-1-carbonyl) phenyl) prop-2-ynyl) - N -phenylpentanamide a) to give ( 35 mg, 56% yield).
  • N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) obtained in Example 2 and potassium carbonate ( 51 mg, 0.372 mmol) was dissolved in N , N -dimethylformamide (DMF) (2 ml) and then stirred at room temperature for 5 minutes. Propargyl bromide (12 ⁇ l, 0.145 mmol) was added to the mixture and stirred at room temperature for 17 hours.
  • N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide 50 mg, 0.124 mmol
  • trimethylsilyl cya obtained in Example 2 above
  • Nide 49 ⁇ l, 0.372 mmol
  • acetonitrile 2 ml
  • Sodium hypochlorite 43 ⁇ l, 0.620 mmol
  • the reaction solution was cooled to room temperature, filtered to remove solids, and then concentrated under reduced pressure.
  • Step 1 N -(3- Fluorophenyl ) Pentaneamide ( N -(3- fluorophenyl ) pentanamide Manufacturing
  • Step 2 tert - Butyl 4- (4- (3- ( N -(3- Fluorophenyl ) Pentanamido ) Prof -One- Inil ) Benzoyl) piperazine-1-carboxylate ( tert -butyl 4- (4- (3- ( N Preparation of-(3-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
  • N- (3-fluorophenyl) pentaneamide (101 mg, 0.519 mmol) obtained in step 1 and tert -butyl 4- (4- (3- (methylsulfonyl) obtained in step 4 of Example 1 Oxy) prop-1-ynyl) benzoyl) piperazine-1-carboxylate (329 mg, 0.779 mmol) in the same manner as in step 5 of Example 1 above, the final product tert - butyl 4- (4- (3- ( N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate ( tert -butyl 4- (4- (3- ( N- (3 -fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (197 mg, 73% yield).
  • Example 14 A N obtained in Example 14 - (3-fluorophenyl) - N - (3- (4- (piperazine-1-carbonyl) phenyl) prop-2-ynyl), pentane amide (46 mg, 0.109 mmol) and 2-iodopropane (0.375 ml, 3.77 mmol) to give the final product N- (3-fluorophenyl) -N- (3- (4- (4-isopropylpipepe) piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide (N - (3-fluorophenyl) - N - (3- (4- (4-isopropylpiperazine-1-carbonyl) prop-2- phenyl) ynyl) pentanamide) was obtained (20 mg, 40% yield).
  • Step 1 N - (4-fluorophenyl) pentaneamide ( N -(4-fluorophenyl) pentanamide Manufacturing
  • Step 2 tert -Butyl 4- (4- (3- ( N -(4- Fluorophenyl ) Pentanamido ) Prof -One- Inil ) Benzoyl) piperazine-1-carboxylate ( tert -butyl 4- (4- (3- ( N Preparation of-(4-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
  • Example 17 The embodiment one N obtained in Example 17 - (4-fluorophenyl) - N - (3- (4- (piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide (1.06 g, 2.51 mmol) and 2-iodopropane (0.375 ml, 3.77 mmol) in the same manner as in Example 15 above, where the final product was N- (4-fluorophenyl) -N- (3- ( 4- (4-isopropyl-piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentane amide (N - (4-fluorophenyl) - N - (3- (4- (4-isopropylpiperazine-1- carbonyl) phenyl) prop-2-ynyl) pentanamide) was obtained (465 mg, 40% yield).
  • Step 1 (4-bromophenyl) (morpholino) methanone ((4-bromophenyl) (morpholino) methanone)
  • Step 2 (4- (3-hydroxyprop-1-ynyl) phenyl) (morpholino) methanone)
  • Example 4 was prepared using (4-bromophenyl) (morpholino) methanone (455 mg, 1.68 mmol) and propagyl alcohol (0.196 ml, 3.36 mmol) obtained in step 1. (4- (3-hydroxyprop-1-ynyl) phenyl) (morpholino) methanone ((4- (3-hydroxyprop-1-ynyl) phenyl) (morpholino) methanone) was obtained (371 mg, yield 90%).
  • step 3 3 -(4-( Morpholine -4- Carbonyl ) Phenyl) Prof -2- Inil Methanesulfonate Preparation of (3- (4- (morpholine-4-carbonyl) phenyl) prop-2-ynyl methanesulfonate)
  • Step 4 N -(3- (4- ( Morpholine -4- Carbonyl ) Phenyl) Prof -2- Inil )- N - Phenylpentane amy De ( N -(3- (4- (morpholine-4-carbonyl) phenyl) prop-2-ynyl)- N -phenylpentanamide)
  • N -phenylpentaneamide (143 mg, 0.81 mmol) obtained in step 1 of Example 1 and 3- (4- (morpholine-4-carbonyl) phenyl) prop-2- obtained in step 3 above.
  • Example 20 N -Phenyl- N -(3- (4- (piperidine-1- Carbonyl ) Phenyl) Prof -2- Inil ) Pentaneamide ( N -phenyl- N -(3- (4- ( piperidine Preparation of -1-carbonyl) phenyl) prop-2-ynyl) pentanamide) (LMT-683)
  • Step 1 (4-bromophenyl) (piperidin-1-yl) methanone ((4- bromophenyl ) ( piperidin -1-yl) methanone))
  • Step 2 (4- (3- Hydroxyprop -One- Inil ) Phenyl) pyridin-1-yl) Metanon Preparation of ((4- (3-hydroxyprop-1-ynyl) phenyl) (piperidin-1-yl) methanone)
  • step 3 3 -(4- (pyridine-1- Carbonyl ) Phenyl) Prof -2- Inil Methanesulfonate Preparation of (3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl methanesulfonate)
  • Step 4 N -Phenyl- N -(3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide ( N -phenyl- N Preparation of-(3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide)
  • N -phenylpentanamide (146 mg, 0.82 mmol) obtained in step 1 of Example 1 and 3- (4- (pyridine-1-carbonyl) phenyl) prop-2-ynyl obtained in step 3 above.
  • Prop-2-ynyl) pentaneamide N- phenyl- N- (3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide
  • step 1 4 - Bromo - N , N - Diethylbenzamide (4- bromo - N , N -diethylbenzamide)
  • Step 2 N , N - Diethyl -4- (3-hydroxyprop-1-ynyl) benzamide ( N , N Preparation of -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide)
  • N , N -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide was obtained.
  • N , N- diethyl-4- (3-hydroxyprop-1-ynyl) benzamide (425 mg, 67% yield).
  • step 3 3 -(4-( Diethylcarbamoyl ) Phenyl) Prof -2- Inil Methanesulfonate Preparation of (3- (4- (diethylcarbamoyl) phenyl) prop-2-ynyl methanesulfonate)
  • N -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide (425 mg, 1.84 mmol) and methanesulfonyl chloride (0.16 ml, 2.02 mmol) obtained in step 2 above.
  • 3- (4- (diethylcarbamoyl) phenyl) prop-2-ynyl methanesulfonate (3- (4- (diethylcarbamoyl) phenyl) prop-2-ynyl in the same manner as in Step 4 of Example 1 above methanesulfonate) was obtained (483 mg, 85% yield).
  • Step 4 N , N - Diethyl -4- (3- ( N -Phenylpentaneamido) prop-1-ynyl) benzamy De ( N , N -diethyl-4- (3- ( N -phenylpentanamido) prop-1-ynyl) benzamide)
  • N -phenylpentanamide (94 mg, 0.530 mmol) obtained in step 1 of Example 1 and 3- (4- (diethylcarbamoyl) phenyl) prop-2-ynyl obtained in step 3 above.
  • methanesulfonate (246 mg, 0.795 mmol) in the same manner as in Step 5 of Example 1, the final product, N , N -diethyl-4- (3- ( N -phenylpentaneamido) prop- 1-ynyl) benzamide ( N , N- diethyl-4- (3- ( N- phenylpentanamido) prop-1-ynyl) benzamide) was obtained (81 mg, 39% yield).
  • Example 22 N -Phenyl- N -(3- (3- (piperazin-1- Carbonyl ) Phenyl) Prof -2- Inil ) Pentaneamide ( N -phenyl- N -(3- (3- ( piperazine Preparation of -1-carbonyl) phenyl) prop-2-ynyl) pentanamide) (LMT-837)
  • Step 1 tert -Butyl 4- (3- Bromobenzoyl Piperazine-1- Carboxylate ( tert -butyl Preparation of 4- (3-bromobenzoyl) piperazine-1-carboxylate)
  • Step 2 tert -Butyl 4- (3- (3- Hydroxyprop -One- Inil ) Benzoyl Piperazine-1- Carboxylate ( tert -butyl 4- (3- (3- hydroxyprop -One- ynyl ) benzoyl ) piperazine -1-carboxylate)
  • Step 3 tert -Butyl 4- (3- (3- ( Methylsulfonyloxy ) Prof -One- Inil ) Benzoyl Piperazine-1-carboxylate ( tert -butyl 4- (3- (3- ( methylsulfonyloxy ) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
  • Step 4 tert -Butyl 4- (3- (3- ( N - Phenylpentaneamido ) Prof -One- Inil ) Benzoyl Piperazine-1-carboxylate ( tert -butyl 4- (3- (3- ( N - phenylpentanamido ) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
  • N -phenylpentaneamide (275 mg, 1.55 mmol) obtained in step 1 of Example 1 and tert -butyl 4- (3- (3- (methylsulfonyloxy) prop-1 obtained in step 3) -Inyl) benzoyl) piperazine-1-carboxylate (982 mg, 2.33 mmol) in the same manner as in Step 5 of Example 1, tert -butyl 4- (3- (3- ( N- Phenylpentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate ( tert -butyl 4- (3- (3- ( N -phenylpentanamido) prop-1-ynyl) benzoyl) piperazine-1- carboxylate) (323 mg, 70% yield).
  • Step 5 N -Phenyl- N -(3- (3- (piperazin-1- Carbonyl ) Phenyl) Prof -2- Inil ) Pentaneamide ( N -phenyl- N -(3- (3- ( piperazine -1-carbonyl) phenyl) prop-2- ynyl ) pentanamide ) (LMT-837)
  • N -phenyl- N- (3- (3- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) obtained in Example 22 and formaldehyde ( 37% in H 2 O) (1.5 ml) was dissolved in formic acid (2.0 ml), heated at 100 ° C., refluxed and stirred for 4 hours. The reaction solution was concentrated under reduced pressure, and then titrated by addition of aqueous sodium hydroxide solution (2.0 M). Thereafter, the mixture was diluted with dichloromethane and washed with water and brine.
  • Example 24 N -(3- (3- (4- Isopropyl piperazine -One- Carbonyl ) Phenyl) Prof -2- Inil )- N -Phenylpentaneamide ( N -(3- (3- (4- isopropylpiperazine -1-carbonyl) phenyl) prop-2-ynyl)- N -phenylpentanamide) (LMT-842)
  • N -phenyl- N- (3- (3- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (152 mg, 0.36 mmol) obtained in Example 22 and 2-Io
  • Example 25 tert -Butyl-4- (3- (3- ( N -(4- Fluorophenyl ) Pentanamido ) Prof -One- Inil ) Benzoyl) piperazine-1-carboxylate ( tert -butyl 4- (3- (3- ( N -(4-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) ( LMT -887)
  • N- (4-fluorophenyl) pentaneamide (275 mg, 1.41 mmol) obtained in Step 1 of Example 16 and tert -butyl 4- (3- (3- (Methylsulfonyloxy) prop-1-ynyl) benzoyl) piperazine-1-carboxylate (894 mg, 2.12 mmol) in the same manner as in Step 5 of Example 1 above, the final product tert -butyl- 4- (3- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazin-1-carboxylate ( tert -butyl 4- (3- (3- ( N- (4-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (625 mg, 85% yield).
  • Example 26 N -(4- Fluorophenyl )- N -(3- (3- (piperazin-1- Carbonyl ) Phenyl) Prof -2-ynyl) pentaneamide ( N -(4- fluorophenyl )- N -(3- (3- ( piperazine Preparation of -1-carbonyl) phenyl) prop-2-ynyl) pentanamide) (LMT-888)
  • Example 27 N -(4- Fluorophenyl )- N -(3- (3- (4- Isopropyl piperazine -One- Carbonyl ) Phenyl) prop-2-ynyl) pentaneamide ( N -(4- fluorophenyl )- N -(3- (3- (4-isopropylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) ( LMT - 889) Produce
  • Example 26 A N obtained in Example 26 - (4-fluorophenyl) - N - (3- (3- (piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide (150 mg, 0.356 mmol) and 2-iodopropane (53 ⁇ l, 0.534 mmol) in the same manner as in Example 5, where the final product, N- (4-fluorophenyl) -N- (3- ( 3- (4-isopropylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide was obtained (66 mg, 40% yield).
  • Step 1 N -(Prop-2-ynyl) aniline ( N Preparation of-(prop-2-ynyl) aniline)
  • Step 2 N -Phenyl- N - (Prop-2-ynyl) pentaneamide ( N -phenyl- N Preparation of-(prop-2-ynyl) pentanamide)
  • Step 3 N -(3- (4- Hydroxyphenyl ) Prof -2- Inil )- N - Phenylpentaneamide ( N -(3- (4-hydroxyphenyl) prop-2-ynyl)- N -phenylpentanamide)
  • Triethyl N -phenyl- N- (prop-2-ynyl) pentaneamide (550 mg, 2.55 mmol) and 4-iodophenol (422 mg, 1.92 mmol) obtained in step 2 were obtained. Dissolved in amine (15 ml) and stirred for 5 minutes. Bis (triphenylphosphine) palladium (II) dichloride (89 mg, 0128 mmol) and copper iodide (I) (49 mg, 0.255 mmol) were added to the mixture, heated at 50 ° C., refluxed and stirred for 5 hours. It was.
  • the reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure.
  • Step 1 ethyl 2- (4- (3- ( N - Phenylpentaneamido ) Prof -One- Inil ) Phenoxy Acetate (ethyl (2- (4- (3- ( N -phenylpentanamido) prop-1-ynyl) phenoxy) acetate)
  • N- (3- (4-hydroxyphenyl) prop-2-ynyl) -N -phenylpentanamide (590 mg, 1.92 mmol) and potassium carbonate (796 mg, 5.76 mmol) obtained in Example 28 were prepared. It was dissolved in acetonitrile (15 ml) and stirred for 30 minutes. Ethyl bromoacetate (0.23 ml, 2.11 mmol) was added to the mixture, followed by stirring at room temperature for 17 hours. The reaction solution was filtered to remove solids, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine.
  • step 2 2 -(4- (3- ( N - Phenylpentaneamido ) Prof -One- Inil ) Phenoxy Acetic acid (2- (4- (3- ( N -phenylpentanamido) prop-1-ynyl) phenoxy) acetic acid)
  • step 1 The ethyl 2- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) phenoxy) acetate (530 mg, 1.35 mmol) obtained in step 1 was dissolved in ethanol (15 ml). , And stirred at room temperature for 5 minutes. To the mixture was added 2N sodium hydroxide (NaOH) (0.50 ml), heated at 80 ° C., refluxed and stirred for 3 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine.
  • NaOH sodium hydroxide
  • Example 30 tert -Butyl 4- (5- (3-(( N - Phenylpentaneamido ) Prof -1-yn-1-yl) Picolinoyl Piperazine-1-carboxylate ( tert -butyl 4- (5- (3- (N- phenylpentanamido ) prop-1-yn-1-yl) picolinoyl) piperazine-1-carboxylate) (LMT-834)
  • N -phenyl- N- (prop-2-ynyl) pentaneamide and tert -butyl 4- (5-bromopicolinoyl) piperazine-1-carboxylate (86 mg) obtained in step 2 of Example 28 above , 0.4 mmol) using the same method as in Step 3 of Example 28, the final product tert -butyl 4- (5- (3-(( N -phenylpentaneamido) prop-1-yn-1 -Yl) picolinoyl) piperazine-1-carboxylate (tert-butyl 4- (5- (3- (N-phenylpentanamido) prop-1-yn-1-yl) picolinoyl) piperazine-1-carboxylate) Obtained (36 mg, 35% yield).
  • Example 31 N -Phenyl- N -(3- (6- (piperazin-1- Carbonyl Pyridin-3-yl) Prof -2-yn-1-yl) pentaneamide ( N -phenyl-N- (3- (6- ( piperazine -1-carbonyl) pyridin -3- yl ) prop-2-yn-1-yl) pentanamide) (LMT-835)
  • Example 32 N -(3- (6- Isopropyl piperazine -One- Carbonyl Pyridin-3-yl) Prof -2-yn-1 yl) pentaneamide (N- (3- (6- (4- isopropylpiperazine -1-carbonyl) pyridin Preparation of -3-yl) prop-2-yn-1-yl) -N-phenylpentanamide) (LMT-836)
  • 0.14 mmol) and sodium hydrogencarbonate (27 mg, 0.316 mmol) were cooled in ice, and N , N -dimethylformamide (DMF) (2 ml) was added thereto, followed by stirring for 1 hour.
  • 2-iodopropane (30 ⁇ l, 0.316 mmol) was added to the mixture, ice was removed, and the mixture was heated and refluxed at 60 ° C. for 24 hours.
  • the reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure.
  • step 1 4 - Bromo - N, N - Diethylbenzamide (4- bromo - N, N - diethylbenzamide Manufacturing
  • 4-Bromobenzoic acid (4-gromobenzoic acid) (5.00 g, 24.9 mmol) was dissolved in N, N -dimethylformamide (100.00 ml) and then diisopropylamine (13 ml, 74.6 mmol) was added. To the mixture was added 1-hydroxybenzotriazole hydrate (7.15 mg, 37.30 mmol) and 1-ethyl-3- (3-dibutylaminopropyl) calvoimide hydrochloride (1-Ethyl -3- (3-dimethylaminopropyl) carbodiimide hydrochloride) (5.04 mg, 37.30 mmol) was added and stirred for 5 minutes.
  • Step 2 N, N - Diethyl -4- (3- Hydroxyprop -One- Inil ) Benzamide ( N, N Preparation of -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide)
  • the reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure.
  • step 3 3 -(4-( N, N - Diethylcarbamoyl ) Phenyl) Prof -2- Inil Methanesulfonate Preparation of (3- (4- (diethylcarbamoyl) phenyl) prop-2-yn-1-yl methanesulfonate)
  • N, N -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide (5.16 mg, 22.3 mmol) was dissolved in dichloromethane (100 ml) and cooled on ice. Triethylamine (4.80 ml, 34.4 mmol) was added to the mixture, which was then stirred for 5 minutes. Methanesulfonyl chloride (1.95 ml, 25.2 mmol) was added at the same temperature, ice was removed, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine.
  • Step 4 N, N - Diethyl -4- (3- (N- (3- Fluorophenyl Pentaamido) Prof -1-yn-1-yl) benzamide ( N, N -diethyl-4- (3- ( N -(3- fluorophenyl ) pentanamido prop-1- yn -1-yl) benzamide)
  • N- (3-fluorophenyl) pentaneamide 200 mg, 1.02 mmol was dissolved in tetrahydrofuran (10.00 ml) and cooled on ice. Sodium hydride (73 mg, 3.06 mmol) was added to the mixture, followed by stirring for 1 hour. At the same temperature, 3- (4- ( N, N -diethylcarbamoyl) phenyl) prop-2-ynyl methanesulfonate (475 mg, 1.54 mmol) obtained in step 3 was added thereto, and the mixture was allowed to stand at room temperature. Stir for 4 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine.
  • Example 34 N, N -Diethyl-4- (3- ( N -(4-fluorophenyl) pentamido) prop-1-yn-1-yl) benzamide ( N, N -diethyl-4- (3- ( N -(4-fluorophenyl) pentanamido) prop-1-yn-1-yl) benzamide) (LMT-927)
  • N- (4-fluorophenyl) pentaneamide 200 mg, 1.02 mmol
  • 3- (4- ( N, N -diethylcarbamoyl) phenyl) prop-2-ynyl methanesulfonate (3- (4 -(diethylcarbamoyl) phenyl) prop-2-yn-1-yl methanesulfonate) using N, N-diethyl-4- (3- (N- (4- Fluophenyl) pentaamido) prop-1-yn-1-yl) benzamide
  • N, N -diethyl-4- (3- ( N- (4-fluorophenyl) pentanamido) prop-1-yn-1 -yl) benzamide was obtained (291.7 mg, 70% yield).
  • Example 35 N -(3- (4- ( N, N - Diethylsulfamoyl ) Phenyl) Prof -2- Inil )- N - Phenylpentamide ( N -(3- (4- ( N, N - diethylsulfamoyl ) phenyl) prop-2- ynyl )- N -phenylpentanamide) (LMT-946)
  • step 1 4 - Bromo - N, N - Diethylbenzenesulfonamide (4- bromo - N, N -diethylbenzenesulfonamide)
  • Step 2 N, N - Diethyl -4- (3- Hydroxyprop -One- Inil ) Benzenesulfonamide ( N, N Preparation of -diethyl-4- (3-hydroxyprop-1-ynyl) benzenesulfonamide)
  • the reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure.
  • step 3 3 -(4-( N, N - Diethylsulfamoyl ) Phenyl) Prof -2- Inil Methanesulfonate (3- (4- ( N, N -diethylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate)
  • N, N -diethyl-4- (3-hydroxyprop-1-ynyl) benzenesulfonamide (273.00 mg, 1.02 mmol) obtained in step 2 was dissolved in dichloromethane (10 ml), and then dried on ice. Cooled. Triethylamine (0.21 ml, 1.53 mmol) was added to the mixture, which was then stirred for 5 minutes. Methanesulfonyl chloride (0.09 ml, 1.12 mmol) was added at the same temperature, ice was removed, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine.
  • step 4 4 -(3- Bromoprop -One- Inil )- N, N - Diethylbenzenesulfonamide (4- (3-bromoprop-1-ynyl)- N, N -diethylbenzenesulfonamide)
  • step 3 3- (4- ( N, N -diethylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate (260.00 mg, 0.75 mmol) obtained in step 3 was added to tetrahydrofuran (20.00 ml). It was dissolved in and cooled on ice. Lithium bromide (196.28 mg, 2.26 mmol) was added to the mixture at the same temperature, followed by stirring for 4 hours. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered, and concentrated under reduced pressure.
  • MgSO 4 anhydrous magnesium sulfate
  • the concentrate was purified by a filter to afford 4- (3-bromoprop-1-ynyl) -N, N -diethylbenzenesulfonamide (4- (3-bromoprop-1-ynyl) -N, N -diethylbenzenesulfonamide). Obtained (240.00 mg, 97%)
  • Step 5 N, N - Diethyl -4- (3- ( Phenylamino ) Prof -One- Inil ) Benzenesulfonamide ( N, N Preparation of -diethyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide)
  • Step 6 N -(3- (4- ( N, N - Diethylsulfamoyl ) Phenyl) Prof -2- Inil )- N - Phenylpentamide ( N -(3- (4- ( N, N - diethylsulfamoyl ) phenyl) prop-2- ynyl )- N -phenylpentanamide)
  • N, N -diethyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide (174.00 mg, 0.51 mmol) obtained in step 5 was dissolved in dichloromethane (15.00 ml). Cooled on ice. Triethylamine (0.14 ml, 1.02 mmol) was added to the mixture, which was then stirred for 5 minutes. Valeroyl chloride (0.06 ml, 0.53 mmol) was added at the same temperature, ice was removed, and the mixture was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine.
  • Example 36 N -(3- (4- ( N - Isopropyl sulfamoyl ) Phenyl) Prof -2- Inil )- N - Phenylpentamide ( N -(3- (4- ( N - isopropylsulfamoyl ) phenyl) prop-2- ynyl )- N -phenylpentanamide) (LMT-947)
  • step 1 4 - Bromo - N - Isopropylbenzenesulfonamide (4- bromo - N -isopropylbenzenesulfonamide)
  • step 2 4 -(3- Hydroxyprop -One- Inil )- N - Isopropylbenzenesulfonamide (4- (3-hydroxyprop-1-ynyl)- N -isopropylbenzenesulfonamide)
  • 4- (3-hydroxyprop-1-ynyl) -N -isopropylbenzenesulfonamide (4- (3-hydroxyprop-1-ynyl) -N- isopropylbenzenesulfonamide) was obtained (420.00 mg, 92% yield).
  • step 3 3 -(4-( N - Isopropyl sulfamoyl ) Phenyl) Prof -2- Inil Methanesulfonate (3- (4- ( N -isopropylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate)
  • step 2 4- (3-hydroxyprop-1-ynyl) -N -isopropylbenzenesulfonamide (410.00 mg, 1.62 mmol) and methanesulfonyl chloride (0.14 ml, 1.78 mmol) obtained in step 2;
  • step 3 of Example 35 using 3- (4- ( N -isopropylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate (3- (4- ( N- isopropylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate) was obtained (330.00 mg, 61% yield).
  • step 4 4 -(3- Bromoprop -One- Inil )- N - Isopropylbenzenesulfonamide (4- (3-bromoprop-1-ynyl)- N -isopropylbenzenesulfonamide)
  • Step 5 N - Isopropyl -4- (3- ( Phenylamino ) Prof -One- Inil ) Benzenesulfonamide ( N Preparation of -isopropyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide)
  • Step 6 N -(3- (4- ( N - Isopropyl sulfamoyl ) Phenyl) Prof -2- Inil )- N - Phenylpentaneamide ( N -(3- (4- ( N - isopropylsulfamoyl ) phenyl) prop-2- ynyl )- N -phenylpentanamide)
  • N -isopropyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide (150.00 mg, 0.46 mmol) and valeroyl chloride (0.06 ml, obtained in step 2) 0.48 mmol) to the same method as in step 6 of example 35 using the N - (3- (4- (N - isopropyl-sulfamoyl) phenyl) prop-2-ynyl) - N - phenyl pentane amide (N -(3- (4- ( N- isopropylsulfamoyl) phenyl) prop-2-ynyl) -N- phenylpentanamide) was obtained (94.00 mg, 49.5% yield).
  • Step 2 N -Phenyl- N - (Pro-2yn-1-yl) pentaneamide ( N -phenyl- N -(prop-2- yn -1-yl) pentanamide)
  • N -phenylpentaneamide (19.10 g, 107.40 mmol) obtained in step 1 was dissolved in N, N -dimethylformamide (DMF) (100 ml), the reaction system was nitrogen-substituted, and sodium hydride (5.20). g, 214.80 mmol) was added below freezing and then stirred for 2 hours. Propagyl bromide (18.10 ml, 214.80 mmol) was added to the mixture, followed by stirring for 2 hours at -40 ° C. Water was added to the reaction solution below freezing, diluted with ethyl acetate, and washed with water and brine.
  • DMF N, N -dimethylformamide
  • Step 4 tert -Butyl 4- (3- ( N Phenylpentane amido) prop -1-yn-1-yl) Benzoa ( tert -butyl 4- (3- ( N -phenylpentanamido) prop-1-yn-1-yl) benzoate)
  • Example 38 4- (3- ( N -Phenylpentaneamido) prop-1-pin-1-yl) benzoic acid Preparation of (4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzoic acid) (LMT-1013)
  • Example 38 4- (3- (N-phenylpentaneamido) prop-1-pin-1-yl) benzoic acid obtained in Example 38 (4- (3- (N-phenylpentanamido) prop-1-yn-1 -yl) benzoic acid) (80.00 mg, 0.24 mmol) was dissolved in N, N-dimethylformamide (DMF) (0.70 ml) and stirred in triethylamine (0.70 ml) for 1 hour.
  • DMF N, N-dimethylformamide
  • Example 38 4- (3- (N-phenylpentaneamido) prop-1-pin-1-yl) benzoic acid obtained in Example 38 (4- (3- (N-phenylpentanamido) prop-1-yn-1 -yl) benzoic acid) (80.00 mg, 0.24 mmol) using N, N-dimethylethane-1,2-diamine (0.04 ml, 0.36 mmol) N- (2- (diethylamino) ethyl) -4- (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzamide (N- (2- (dimethylamino) ethyl) -4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamide) was obtained (71.72 mg, 74% yield).
  • Example 41 Ethyl 2- (4- (3- (N- Phenylpentaneamido ) Prof -1-yn-1-yl) Benzamido Acetate (ethyl 2- (4- (3- (N-) phenylpentanamido prop-1- yn -1-yl) benzamido) acetate) (LMT-1014)
  • Example 38 4- (3- (N-phenylpentaneamido) prop-1-pin-1-yl) benzoic acid obtained in Example 38 (4- (3- (N-phenylpentanamido) prop-1-yn-1 -yl) benzoic acid) (150.00 mg, 0.45 mmol) using glycineethylester hydrochloride (93.70 mg, 0.67 mmol) in the same manner as in Example 39 to obtain ethyl 2- (4- (3- ( N-phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetate (ethyl 2- (4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamido) acetate) was obtained (102.18 mg, 54% yield).
  • Ethyl 2- (4- (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetate obtained in Example 41 (ethyl 2- (4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamido) acetate) (46.20 mg, 0.10 mmol) and a 2-mol aqueous solution of sodium hydroxide (0.07 ml, 0.14 mmol) were dissolved in methanol (0.1 ml). Stirred at room temperature for 30 minutes. The acidity of the reaction solution was increased using hydrochloric acid, diluted with ethyl acetate, and washed with water and brine.
  • Example 43 methyl 2- (4- (3- (N- Phenylpentaneamido ) Prof -1-yn-1-yl) Benzamido Propanoate (methyl 2- (4- (3- (N-) phenylpentanamido prop-1- yn -1-yl) benzamido) propanoate) (LMT-1018)
  • Example 38 4- (3- (N-phenylpentaneamido) prop-1-pin-1-yl) benzoic acid obtained in Example 38 (4- (3- (N-phenylpentanamido) prop-1-yn-1 L-alaninemethylester hydrochloride (83.70 mg, 0.60 mmol) in -yl) benzoic acid) (100.00 mg, 0.30 mmol) in the same manner as in Example 39.
  • Step 1 N- (3- Fluorophenyl ) Pentaneamide (N- (3- fluorophenyl ) pentanamide Manufacturing
  • N- (3-fluorophenyl) pentanamide (N- (3-fluorophenyl) pentanamide in the same manner as in Step 1 of Example 37 using 3-fluoroaniline (200.00 mg, 1.79 mmol) ) was obtained (349.00 mg, 99% yield).
  • Step 2 N- (3- FluoroPetyl ) -N- ( Prof -2- Inil ) Pentaneamide Preparation of (N- (3-fluorophenyl) -N- (prop-2-ynyl) pentanamide)
  • N- (3-fluorophenyl) pentaneamide 400.00 mg, 2.05 mmol
  • Potassium hydroxide 230.61 mg, 4.11 mmol
  • tetrabutylammonium iodide tetrabutyl ammonium Iodide
  • Tetrahydrofuran 20.00 ml
  • propargyl bromide (0.19 ml, 2.30 mmol) was added to the mixture, followed by stirring for 20 hours.
  • the reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine.
  • Step 3 N- (3- Fluorophenyl ) -N- (3- (4- Hydroxyphenyl ) Prof -2- Inil ) Pentanah Amide (N- (3- fluorophenyl ) -N- (3- (4- hydroxyphenyl prop-2- ynyl ) pentanamide Manufacturing
  • N- (3-fluorofetyl) -N- (prop-2-ynyl) pentaneamide (270.00 mg, 1.16 mmol) and 4-iodophenol (127.60 mg, obtained in step 2) 0.58 mmol) with N- (3-fluorophenyl) -N- (3- (4-hydroxyphenyl) prop-2-ynyl) pentaneamide (N- (3-fluorophenyl) -N- (3 -(4-hydroxyphenyl) prop-2-ynyl) pentanamide) was obtained (128.00 mg, 67.8% yield).
  • Step 4 ethyl 2- (4- (3- (N- (3- Fluorophenyl ) Pentanamido ) Prof -One- Inil ) Phenoxy Acetate (ethyl 2- (4- (3- (N- (3- fluorophenyl ) pentanamido ) prop-1-ynyl) phenoxy) acetate)
  • step 5 2 -(4- (3- (N- (3- Fluorophenyl ) Pentanamido ) Prof -One- Inil ) Phenoxy ) Acetic acid Preparation of (2- (4- (3- (N- (3-fluorophenyl) pentanamido) prop-1-ynyl) phenoxy) acetic acid)
  • Sodium hydroxide 2M (0.30 ml) was added to the mixture, and the mixture was heated and refluxed at 80 ° C. for 3 hours.
  • the reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine.
  • Step 1 N- (4- Fluorophenyl ) Pentaneamide (N- (4- fluorophenyl ) pentanamide Manufacturing
  • Step 2 N- (4- Fluorophenyl ) -N- ( Prof -2- Inil ) Pentaneamide Preparation of (N- (4-fluorophenyl) -N- (prop-2-ynyl) pentanamide)
  • Step 3 N- (4- Fluorophenyl ) -N- (3- (4- Hydroxyphenyl ) Prof -2- Inil ) Pentanah Amide (N- (4- fluorophenyl ) -N- (3- (4- hydroxyphenyl prop-2- ynyl ) pentanamide Manufacturing
  • N- (4-fluorophenyl) -N- (prop-2-ynyl) pentaneamide (300.00 mg, 1.29 mmol) and 4-iodophenol (140.80 mg, obtained in step 3)
  • N- (4-fluorophenyl) -N- (3- (4-hydroxyphenyl) prop-2-ynyl) pentaneamide N- (4-fluorophenyl) -N- (3 -(4-hydroxyphenyl) prop-2-ynyl) pentanamide
  • Step 4 ethyl 2- (4- (3- (N- (4- Fluorophenyl ) Pentanamido ) Prof -One- Inil ) Phenoxy Acetate (ethyl 2- (4- (3- (N- (4- fluorophenyl ) pentanamido ) prop-1-ynyl) phenoxy) acetate)
  • step 5 2 -(4- (3- (N- (4- Fluorophenyl ) Pentanamido ) Prof -One- Inil ) Phenoxy ) Acetic acid Preparation of (2- (4- (3- (N- (4-fluorophenyl) pentanamido) prop-1-ynyl) phenoxy) acetic acid)
  • cells without BLT2 expression and cells with BLT2 expression were prepared in the following manner.
  • CHO cells were obtained from the Korea Cell Line Bank (KCLB, 10061), which contained 10% FBS (fetal bovine serum; Life technologies, Inc.), penicillin (50 units / mL), and antibiotic antimycotic solution (Life technologies, Inc.). It was incubated at 37 °C, 5% CO 2 conditions in RPMI 1640 medium (Invitrogen) containing the. The cells were maintained in the growth phase by splitting with Trypsin-EDTA for 3 days, respectively, with PBS (phosphate-buffered saline; 137 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 2m MKH 2 PO 4 ) And then added to fresh medium to prepare cells without BLT2 expression.
  • FBS fetal bovine serum
  • penicillin 50 units / mL
  • antibiotic antimycotic solution Life technologies, Inc.
  • CHO-K1 cells were transformed with pcDNA3-long form BLT2 encoding HA-tagged human BLT2, and 0.4 mg / ml of G418 (Invitrogen). , Carlsbad, CA, USA).
  • G418 Invitrogen
  • the selected clones were analyzed by RT-PCR using human-specific BLT2 primers, and representative clones were used in the experiments with cells expressing BLT2 (CHO-BLT2).
  • LTB 4 which is a ligand of BLT2 to BLT2-expressing cells (CHO-BLT2 cells) (300 nM) treated (DMSO +) and ethanol treated (DMSO-) cell growth increased from 20% to 35%, and in BLT2 expressed cells (CHO-BLT2), positive control Phosphorus LY255283 pre-treated, showed about 90% cell growth compared to the control group treated with DMSO, confirming the effect of inhibiting cell growth according to the compound treatment of the Example.
  • the compounds of the present invention can inhibit BLT2-induced cell proliferation with very good efficiency, and the compound can be used as a pharmaceutical ingredient (BLT2-) as a therapeutic agent for the inhibition of anticancer, anti-asthma or other forms of BLT2-related inflammatory diseases. blocking pharmacological molecules).
  • Chemotactic motility was analyzed using a Transwell chamber equipped with a 6.5-mm diameter polycarbonate filter (8- ⁇ m pore size, Corning Costar). Specifically, the bottom surface of the filter was coated with 10 ⁇ g / mL fibronectin in serum free RPMI 1640 medium at 37 ° C. for 1 hour. A dry, coated filter with RPMI 1640 medium containing varying amounts of LTB 4 is placed in the bottom well of the Transwell chamber and finally 2 ⁇ 10 4 cells of CHO cells stably expressing BLT1 and BLT2 in serum-free RPMI 1640 medium. Experiments were performed by loading into the upper wells at / 100 ⁇ L.
  • the cells were pretreated with each inhibitor for 30 minutes before dispensing. After 3 hours of incubation at 37 ° C., 5% CO 2 , the filters were fixed with methanol for 3 minutes and stained with hematoxylin and eosin for 10 minutes.
  • the cells used BLT2-expressing cells (CHO-BLT2 cells) and BLT1-expressing cells (CHO-BLT1), LY255283 and U75302 respectively as a positive control, LTB 4 ligand of BLT2 as a comparative control , (300 nM), LTB 4 , the ligand of BLT1 (10 nM) and LPA (lysophosphatidic acid; 100 nM) were used. Chemotaxis was quantified by counting cells on the lower side of the filter under an optical microscope (magnification, x200). Six fields were counted in each assay, each sample analyzed twice, and the assay was repeated three times.
  • IC 50 50% inhibitory concentration
  • LTB 4 which is a ligand of BLT2 in BLT2-expressing cells (CHO-BLT2 cells)
  • DMSO + DMSO +
  • DMSO- ethanol treated
  • the cell chemotaxis was increased by 2.4 times
  • LY255283 used as a positive control (10 ⁇ M) the ligand LTB Compared to the case of 4 treated, it was confirmed that the coinability of 90%.
  • LTB 4 is a ligand Compared with treatment (DMSO +), chemotaxis was 66%, 90%, and 70.3% inhibited, whereas LTB 4 , a ligand, was expressed in BLT1-expressing cells (CHO-BLT1). It was confirmed that chemotaxis was not suppressed compared to the treatment (DMSO +).
  • LTB 4 and BLT2 binding (ligand binding affinity) inhibition was analyzed by isotope tritium (H3) using the label LTB 4 ([3H] LTB 4, ARC) (specific activity 160.0 Ci / mmol).
  • Experimental method is to put 2 ⁇ 10 6 CHO-BLT2 cells in a 100 mm culture dish and incubate for 48 hours before proceeding. The harvested cells are used five times for one minute in a homogenizer to separate the proteins of the cell membrane. Thereafter, centrifugation was performed for 40 minutes at 45,000 RPM at 4 ° C to harvest only proteins of the cell membrane and quantitate them at a concentration of 40 ⁇ g / 45 ⁇ L.
  • MDA-MB-231 and MDA-MB-453 were intended to confirm the inhibition of the production of free radicals and IL-8 expression in the cells according to the compound treatment of the present invention.
  • MDA-MB-231 cells breast cancer cells, were obtained from Korea Cell Line Bank (Seoul, Korea), and MDA-MB-435 cells were obtained from J.-H. Obtained from Lee (Asan Medical Center, Seoul, Korea). This was 37 ° C. in RPMI 1640 medium (Invitrogen) containing 10% FBS (fetal bovine serum; Life technologies, Inc.), 1% penicillin (50 units / mL), and antibiotic antimycotic solution (Life technologies, Inc.). , And cultured under the condition of 5% CO 2 .
  • FBS fetal bovine serum
  • penicillin 50 units / mL
  • antibiotic antimycotic solution Life technologies, Inc.
  • Intracellular reactive oxygen (H 2 O 2 ) following compound (LMT-696) treatment of the present invention was measured as a function of DCF fluorescence. Specifically, before the measurement of free radicals (ROS) 2 ⁇ 10 5 Cells were grown in 60-mm wells and incubated for 24 hours in RPMI 1640 medium supplemented with FBS. To assess the effect of the compounds of the invention, the cells were pretreated with compound (LMT-696) for 30 minutes each. To measure intracellular free radicals, cells were incubated for 20 minutes in a dark, humid CO 2 incubator at 37 ° C. with H 2 O 2 -sensitive fluorescent substance H 2 DCFDA [Molecular Probes (Eugene, OR)] (20 ⁇ M). It was.
  • H 2 DCFDA is hydrolyzed to DCF in the cell and is present in the cell, and oxidized to DCF showing high fluorescence in the presence of H 2 O 2 , and thus the amount of active oxygen was measured using this property.
  • cells were harvested using trypsin-EDTA and resuspended in serum-free RPMI 1640 without phenol red for confirmation of free radical production in the detector.
  • DCF fluorescence was measured with excitation and emission wavelengths of 488 and 530 nm, respectively, using a FACS Calibur flow cytometer (Becton Dickinson, NJ).
  • RNAs were isolated from cells by Easy Blue (Intron, Sungnam, Korea) and quantified at 260 nm absorbance. RNA (1.25 ⁇ g) was synthesized as cDNA by reverse transcription by polymerase chain reaction (PCR) technique. Expression levels were confirmed using primers that specifically bind to IL-8 and glyceraldehydes-3-phosphate dehydrogenase (GAPDH).
  • PCR polymerase chain reaction
  • BioCoat Matrigel Invasion Chambers (BD Biosciences, Bedford, Mass.) was used to measure the penetration of breast cancer cells following the compound treatment of the present invention.
  • 5 ⁇ 10 4 breast cancer cells were harvested with trypsin-EDTA, then resuspended in RPMI 1640 included in 0.5% serum and transferred to Matrigel inserts.
  • RPMI 1640 containing 5% serum was placed in the lower chamber and the cells were incubated at 37 ° C. for 36 hours.
  • Each filter was fixed with methanol for 3 minutes and stained with hematoxylin and eosin for 10 minutes.
  • Permeability of cancer cells was quantified by the number of cells on the lower side of the filter under an optical microscope (magnification x 200). Six fields were quantified for each assay. Each sample was analyzed twice, and the analysis was repeated three times.
  • the metastasis experiment of breast cancer cells according to the compound treatment of the present invention was approved by Korea University Ethics Committee, and all experimental animals used in this experiment were performed according to the approved guidelines of Korea University Animal Care and Use Committee. Cancer cells were injected into 6-week-old female nude mice (Charles River, Wilmington, Mass.) To confirm cancer cell metastasis. After 24 hours pretreatment of the compound of the present invention (LMT-696, 10 ⁇ M), LY255283, U75302 and DMSO in breast cancer cells, harvested with trypsin-EDTA, resuspended in PBS, and anesthetized with zoletil (50 mg / kg) in mice. 2 x 10 6 breast cancer cells were injected by intraperitoneal injection.
  • metastasis of cancer cells was inhibited 40% by treatment of the compound of the present invention (LMT-696) compared to the control, positive
  • the control group LY255283 was suppressed 36% by treatment, while the U75302 treatment did not inhibit the metastasis of cancer cells MDA-MB-231.
  • the compound of the present invention (LMT-696) can inhibit the production of free radicals and IL-8 in the cells of the cancer cells and consequently inhibit the penetration and metastasis of the cancer cells, a pharmaceutical component having excellent anticancer efficacy It can be used as.
  • mast cells play an important role.
  • mast cells are activated to release various cytokines (interlukin-4 and interlukin-13).
  • the cytokine causes phenomena such as influx of inflammatory cells, mucus production, and airway contraction.
  • the present inventors received female BALB / c mice that were 7 weeks old (18-20 g) from Orient (Seoungnam, Korea) and used them in the experiments to induce asthma.
  • Airway hypersensitivity was measured 24 days after initial sensitization, and mice were dissected on day 25 to observe the asthma phenotype of inflammatory cytokine IL-4 expression and the influx of inflammatory cells (neutrophils).
  • LPS Lipopolysaccharide
  • SVA egg white albumin
  • OVA egg white albumin
  • Airway constrictor administration was sprayed through the inlet of the chamber for 3 minutes using an ultrasonic nebulizer. Airway hypersensitivity was analyzed using enhanced pause as an indicator of asthma. Airway wash cell numbers were quantified by counting cells under an optical microscope (magnification, x 200). Four fields were counted in each assay, each sample analyzed twice, and the assay was repeated three times.
  • airway resistance was increased by about 13 times compared to asthma-induced mouse (OVA + LPS) compared to non-asthma-induced mouse (Normal), and the compound of the present invention (LMT-1013)
  • OVA + LPS asthma-induced mouse
  • Normal non-asthma-induced mouse
  • LMT-1013 the compound of the present invention
  • the present invention relates to a novel compound that exhibits Leukotriene B4 receptor 2 (BLT2) inhibitory activity and a pharmaceutical composition for preventing or treating an inflammatory disease comprising the same.
  • BLT2 Leukotriene B4 receptor 2
  • the present inventors have identified a novel compound that exhibits BTL2 inhibitory activity in order to solve in vivo instability and difficulty in mass production, which are problems of conventional inflammatory disease treatment substances, and has excellent cancer cell death enhancement and metastasis suppression effect and chemotaxis inhibition of the compound.
  • the effects, and anti-asthma effects have been confirmed experimentally, it is expected that it can be usefully used as a pharmaceutical composition for treating inflammatory diseases.

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Abstract

The present invention relates to a novel compound showing leukotriene B4 receptor 2 (BLT2) inhibitory activity and a pharmaceutical composition, for preventing or treating inflammatory diseases, comprising same as an active ingredient. In the present invention, a novel compound showing BLT2 inhibitory activity is investigated and the enhanced cancer cell death, metastasis suppression, chemotaxis inhibition, antiasthmatic effect and the like shown in the compound have been experimentally confirmed. Therefore, the present invention enables more fundamental approach and target treatment for treating or preventing inflammatory diseases.

Description

BLT 저해 활성을 갖는 신규 화합물 및 이를 유효성분으로 포함하는 염증성 질환 예방 또는 치료용 조성물Novel compounds having BLT inhibitory activity and compositions for preventing or treating inflammatory diseases comprising the same as active ingredients
본 발명은 신규 화합물 및 이의 용도에 관한 것으로서, 보다 구체적으로는 BLT2 (Leukotriene B4 receptor 2) 억제 활성을 나타내는 신규 화합물 및 이를 유효성분으로 포함하는 염증성 질환 예방 또는 치료용 약학적 조성물에 관한 것이다.The present invention relates to a novel compound and its use, and more particularly, to a novel compound exhibiting inhibitory activity of Leukotriene B4 receptor 2 (BLT2) and a pharmaceutical composition for preventing or treating inflammatory diseases comprising the same as an active ingredient.
염증반응은 생체나 조직에 물리적 작용이나 화학적 물질, 세균 감염, 면역학적 자극 등이 가해질 때, 이에 대해 방어할 수 있도록 다양한 작용기전을 통해 활성화되는 인체의 면역 시스템 중 하나이다. 다만, 이와 같은 염증반응이 지속되면 오히려 점막 손상을 촉진하게 되며, 이에 따라 발적, 발열, 종창, 동통, 기능장애 등으로 인한 류마티스 관절염, 동맥경화증, 위염, 천식 등의 염증성 질환을 유발하는 것을 알려져 있다. 이러한, 염증반응은 시간의 경과에 따라 급성염증과 만성염증으로 구분되며, 급성염증은 염증반응이 수 일 내지 수 주간 지속되고, 홍반, 발열, 통증, 부종과 같은 증상을 야기하는 반면, 만성염증은 장기간 염증상태가 지속되는 것으로 때로는 수 년 내지 수십 년에 걸쳐 나타나며, 단핵세포의 침윤, 섬유모세포 및 모세혈관의 증식, 결합조직의 증가로 인한 섬유화, 조직의 파괴 등의 조직학적 변화를 수반한다.Inflammatory reactions are one of the body's immune systems that are activated through various mechanisms of action to protect against physical or chemical agents, bacterial infections, and immunological stimuli. However, if such an inflammatory response persists, it promotes mucosal damage, thereby causing inflammatory diseases such as rheumatoid arthritis, arteriosclerosis, gastritis, and asthma due to redness, fever, swelling, pain, and dysfunction. have. These inflammatory reactions are classified into acute and chronic inflammations over time, and acute inflammations can cause symptoms such as erythema, fever, pain, and edema, while inflammatory reactions last several days to several weeks. Is a prolonged inflammatory state, sometimes over several years to decades, and is accompanied by histological changes such as invasion of monocytes, proliferation of fibroblasts and capillaries, fibrosis due to increased connective tissue, and tissue destruction. .
구체적으로, 생체 내 염증성 자극이 가해지면, 국소적으로 히스타민(histamine), 브레디키닌 (bradykinin), 프로스타글란딘 (prostaglandins), 산화질소(nitric oxide, NO), 각종 전염증성 사이토카인 (pro-inflammatory cytokines) 등이 합성, 분비되며, 이들은 혈관 확장 뿐 아니라 홍반, 발열, 통증, 부종을 유발시킨다. 특히, 체내 염증 과정에서 인터페론-감마 (interferon-γ, INF-γ), 종양괴사인자-알파 (tumor necrosis factor-α, TNF-α), 인터루킨-1 (interleukin-1, IL-1), 인터루킨-6(interleukin-6, IL-6) 등의 사이토카인과 같은 일반적인 면역 인자들 외에도 산화질소 (NO) 및 프로스타글란딘 E2 (prostaglandin E2, PGE2)이 주요 염증 유발 물질로 잘 알려져 있다.Specifically, when inflammatory stimuli are applied in vivo, histamine, bradykinin, prostaglandins, nitric oxide (NO), and various pro-inflammatory cytokines (pro-inflammatory cytokines) ) Are synthesized and secreted, and they cause vascular enlargement as well as erythema, fever, pain and edema. In particular, interferon-gamma (INF-γ), tumor necrosis factor-α (TNF-α), interleukin-1 (interleukin-1, IL-1), interleukin during inflammatory processes in the body In addition to common immune factors such as cytokines such as -6 (interleukin-6, IL-6), nitric oxide (NO) and prostaglandin E2 (PGE2) are well known as major inflammatory agents.
종래 염증 반응의 종결은 염증을 시작하는 물질들의 수준이 감소하면 자연적이고 수동적으로 일어나는 현상이라고 알려져 왔으나, Serhan 등이 lipoxin, resovin, protectin 류 등을 발견하여 이들에 의해 염증의 시작에 관여하는 프로스타글란딘 (prostaglandin)류처럼 염증의 종결이 능동적으로 촉진된다는 것을 발견하였다. 예를 들어, Resolvin E1은 통증에 효과적이며, RvE1는 염증의 종결을 유도하여 알레르기 염증 질환의 치료에 효과가 있다는 점이 보고된 바 있다. 또한, 만성 염증 질환에서 이러한 염증 종결을 능동적으로 촉진하는 인자들 즉, lipoxin A4, 아스피린에 의해 유도되는 lipoxin의 수준이 천식 환자 및 죽상동맥경화 환자에서 낮게 관찰됨이 보고된 바 있다. Conventionally, the termination of the inflammatory response has been known to occur naturally and passively when the level of inflammation-inducing substances decreases. However, Serhan et al. Found lipoxin, resovin, protectin, and the like, and prostaglandins are involved in the initiation of inflammation. Like prostaglandins, it has been found that the termination of inflammation is actively promoted. For example, it has been reported that Resolvin E1 is effective in pain and RvE1 is effective in treating allergic inflammatory diseases by inducing the termination of inflammation. In addition, it has been reported that the levels of the factors that actively promote the termination of inflammation in chronic inflammatory diseases, namely lipoxin A4 and aspirin-induced lipoxin, are low in asthma patients and atherosclerosis patients.
이에, 염증의 종결을 유도하는 신규 물질을 발굴하여, 염증 종결 이상과 관련된 질환을 치료하고자 하는 시도가 다양하게 진행되고 있으나 (한국공개특허 10-2015-0011875), lipoxin, resovin 등에 속한다고 알려진 화합물은 그 구조에 여러 개의 이중결합이 포함되어 있어 대사적으로 불안정하여 생체에서 빠르게 분해되는 등의 단점을 가지고 있으며, 물질을 대량생산하여 약으로 개발하기에는 다소 어려워, 약물성에 큰 문제점을 가지고 있다. Therefore, various attempts have been made to discover new substances that induce the termination of inflammation, and to treat diseases related to abnormal termination of inflammation (Korea Patent Publication No. 10-2015-0011875), compounds known to belong to lipoxin, resovin, etc. Since the structure contains a plurality of double bonds, such as metabolic instability and has a disadvantage of rapid decomposition in the living body, and mass production of the material is somewhat difficult to develop into a drug, has a big problem in drug properties.
한편, 류코트리엔 (Leukotriene B4; LTB4)은 급성 및 만성염증을 매개하는 5-리폭시제나제 경로에 의해 아라키돈산(AA)으로부터 합성되는 염증성 리피드 매개체 군이다. LTB4는 BLT1과 BLT2 의 두 가지 형태의 수용체들에 결합함으로써 생물학적 영향을 주는 것으로 알려져 있다. BLT2(Leukotriene B4 receptor 2)는 GPCR(G protein-coupled receptor) 군 중 하나로 LTB4에 대해 낮은 친화력을 갖는 수용체이며, 5-리폭시제나제 의존성 경로를 통해 유도된 아라키돈산(AA)의 리피드 매개체이다. On the other hand, leukotriene B4 (LTB 4 ) is a group of inflammatory lipid mediators synthesized from arachidonic acid (AA) by the 5-lipoxygenase pathway that mediates acute and chronic inflammation. LTB 4 is known to have a biological effect by binding to two types of receptors, BLT1 and BLT2. Leukotriene B4 receptor 2 (BLT2) is a group of G protein-coupled receptors (GPCRs) that has low affinity for LTB 4 and is a lipid mediator of arachidonic acid (AA) induced through a 5-lipoxygenase dependent pathway. to be.
이에, 본 발명자들은 상기와 같은 종래의 문제점을 해결하기 위하여, 보다 효과적인 염증의 종결을 유도하는 물질을 개발하기 위한 연구를 계속하던 중, BLT2 억제 활성을 나타내는 신규 화합물을 제조하였으며, 상기 화합물을 포함하는 염증성 질환 치료제를 최초로 고안하였다.Accordingly, the present inventors, while continuing to research to develop a substance that induces more effective termination of inflammation, in order to solve the conventional problems as described above, has produced a novel compound showing a BLT2 inhibitory activity, including the compound It was the first to develop an inflammatory disease treatment.
본 발명은 상기와 같은 문제점을 해결하기 위해 안출된 것으로서, 본 발명자들은 BLT2 억제 활성을 나타내는 신규 화합물의 염증성 질환 치료 효과를 확인하고 이에 기초하여 본 발명을 완성하게 되었다.The present invention has been made to solve the above problems, the present inventors have confirmed the therapeutic effect of the inflammatory disease of the novel compound showing the BLT2 inhibitory activity and completed the present invention based on this.
이에, 본 발명의 목적은 BLT2 억제 활성을 나타내는 신규 화합물 또는 이의 약학적으로 허용 가능한 염을 제공하는 것이다.Accordingly, it is an object of the present invention to provide novel compounds or pharmaceutically acceptable salts thereof which exhibit BLT2 inhibitory activity.
또한, 본 발명의 다른 목적은 상기 신규 화합물 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는, 염증성 질환 예방 또는 치료용 약학적 조성물을 제공하는 것이다.In addition, another object of the present invention is to provide a pharmaceutical composition for preventing or treating inflammatory diseases, comprising the novel compound or a pharmaceutically acceptable salt thereof as an active ingredient.
그러나 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다.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.
상기와 같은 본 발명의 목적을 달성하기 위하여, 본 발명은 BLT2 억제 활성을 나타내는 신규 화합물 또는 이의 약학적으로 허용 가능한 염을 제공한다.In order to achieve the above object of the present invention, the present invention provides a novel compound or a pharmaceutically acceptable salt thereof exhibiting BLT2 inhibitory activity.
본 발명의 일 구현예로서, 상기 화합물은 tert-부틸 4-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-(4-(4-메틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-에틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-(2-하이드록시에틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-(사이클로프로필메틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-사이클로헥실피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-(사이클로헥실메틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-아이소부틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-페닐-N-(3-(4-(4-(프로프-2-이닐)피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-(4-(4-시아노피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; tert -부틸 4-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; N-(3-플루오로페닐)-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-플루오로페닐)-N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; tert-부틸 4-(4-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; N-(4-플루오로페닐)-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(4-플루오로페닐)-N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-(4-(몰폴린-4-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-페닐-N-(3-(4-(피페리딘-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N,N-다이에틸-4-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤즈아마이드; N-페닐-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-(3-(4-메틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(3-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; tert-부틸-4-(3-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; N-(4-플루오로페닐)-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(4-플루오로페닐)-N-(3-(3-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-(4-하이드록시페닐)프로프-2-이닐)-N-페닐펜탄아마이드; 2-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)페녹시)아세트산; tert-부틸 4-(5-(3-((N-페닐펜탄아미도)프로프-1-인-1-일)피콜리노일)피페라진-1-카복실레이트; N-페닐-N-(3-(6-(피페라진-1-카보닐)피리딘-3-일)프로프-2-인-1-일)펜탄아마이드; N-(3-(6-아이소프로필피페라진-1-카보닐)피리딘-3-일)프로프-2-인-1일)펜탄아마이드; N,N-다이에틸-4-(3-(N-(3-플루오로페닐)펜타아미도)프로프-1-인-1-일)벤즈아마이드; N,N-다이에틸-4-(3-(N-(4-플루오로페닐)펜타아미도)프로프-1-인-1-일)벤즈아마이드; N-(3-(4-(N,N-다이에틸설파모일)페닐)프로프-2-이닐)-N-페닐펜타아마이드; N-(3-(4-(N-아이소프로필설파모일)페닐)프로프-2-이닐)-N-페닐펜타아마이드; tert-부틸 4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤조에이트; 4-(3-(N-페닐펜탄아미도)프로-1-핀-1-일)벤조익산; N-에틸-4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아마이드; N-(2-(다이에틸아미노)에틸)-4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아마이드; 에틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)아세테이트; 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)아세틱산; 메틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)프로파노에이트; 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)프로피오닉산; 2-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세트산; 및 2-(4-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세트산으로 이루어진 군으로부터 선택될 수 있다.In one embodiment of the present invention, the compound is tert -butyl 4- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate; N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide; N- (3- (4- (4-methylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N- (3- (4- (4-ethylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N- (3- (4- (4-isopropylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N- (3- (4- (4- (2-hydroxyethyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N- (3- (4- (4- (cyclopropylmethyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N- (3- (4- (4-cyclohexylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N- (3- (4- (4- (cyclohexylmethyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N- (3- (4- (4-isobutylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N -phenyl- N- (3- (4- (4- (prop-2-ynyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide; N- (3- (4- (4-cyanopiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; tert - butyl 4- (4- (3- ( N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate; N - (3-fluorophenyl) - N - (3- (4- ( piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; N - (3-fluorophenyl) - N - (3- (4- (4- isopropyl-piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; tert -butyl 4- (4- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate; N - (4-fluorophenyl) - N - (3- (4- ( piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; N - (4-fluorophenyl) - N - (3- (4- (4-isopropyl-piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; N- (3- (4- (morpholin-4-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N -phenyl- N- (3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide; N , N -diethyl-4- (3- ( N -phenylpentaneamido) prop-1-ynyl) benzamide; N -phenyl- N- (3- (3- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide; N- (3- (3- (4-methylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N- (3- (3- (4-isopropylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; tert -butyl-4- (3- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate; N - (4-fluorophenyl) - N - (3- (3- ( piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; N - (4- fluorophenyl) - N - (3- (3- (4- isopropyl-piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; N- (3- (4-hydroxyphenyl) prop-2-ynyl) -N -phenylpentanamide; 2- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) phenoxy) acetic acid; tert -butyl 4- (5- (3-(( N -phenylpentaneamido) prop-1-yn-1-yl) picolinoyl) piperazin-1-carboxylate; N -phenyl- N- ( 3- (6- (piperazin-1-carbonyl) pyridin-3-yl) prop-2-yn-1-yl) pentaneamide; N- (3- (6-isopropylpiperazin-1-carbo Yl) pyridin-3-yl) prop-2-yn-1 yl) pentaneamide; N, N -diethyl-4- (3- ( N- (3-fluorophenyl) pentamido) prop-1-yn-1-yl) benzamide; N, N -diethyl-4- (3- ( N- (4-fluorophenyl) pentamido) prop-1-yn-1-yl) benzamide; N- (3- (4- ( N, N -diethylsulfamoyl) phenyl) prop-2-ynyl) -N -phenylpentamide; N- (3- (4- ( N -isopropylsulfamoyl) phenyl) prop-2-ynyl) -N -phenylpentamide; tert -butyl 4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzoate; 4- (3- ( N -phenylpentanamido) pro-1-pin-1-yl) benzoic acid; N -ethyl-4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamide; N- (2- (diethylamino) ethyl) -4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamide; Ethyl 2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetate; 2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetic acid; Methyl 2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) propanoate; 2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) propionic acid; 2- (4- (3- ( N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy) acetic acid; And 2- (4- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy) acetic acid.
본 발명은 상기 신규 화합물 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는, 염증성 질환 예방 또는 치료용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease, comprising the novel compound or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명의 일 구현예로서, 상기 염증성 질환은 천식, 죽상경화증, 암, 피부가려움증, 류마티스 관절염 및 염증성 장 질환으로 이루어진 군으로부터 선택될 수 있다. In one embodiment of the present invention, the inflammatory disease may be selected from the group consisting of asthma, atherosclerosis, cancer, itching of the skin, rheumatoid arthritis and inflammatory bowel disease.
본 발명의 다른 구현예로서, 상기 조성물은 BLT2 (Leukotriene B4 receptor 2) 활성을 저해시킬 수 있다.In another embodiment of the present invention, the composition may inhibit Leukotriene B4 receptor 2 (BLT2) activity.
상기 약학적 조성물을 개체에 투여하는 단계를 포함하는 염증성 질환의 치료방법을 제공한다.It provides a method of treating an inflammatory disease comprising administering the pharmaceutical composition to a subject.
본 발명은 상기 신규 화합물 또는 이의 약학적으로 허용 가능한 염을 포함하는 조성물의 염증성 질환의 치료용도를 제공한다.The present invention provides a therapeutic use of an inflammatory disease of a composition comprising the novel compound or a pharmaceutically acceptable salt thereof.
본 발명은 BLT2 (Leukotriene B4 receptor 2) 억제 활성을 나타내는 신규 화합물 및 이를 포함하는 염증성 질환 예방 또는 치료용 약학적 조성물에 관한 것이다. 본 발명자들은 종래의 염증성 질환 치료 물질의 문제점인 생체 내 불안정성 및 대량 생산의 어려움을 해소하기 위하여 BTL2 억제 활성을 나타내는 신규 화합물을 규명하였으며, 상기 화합물의 우수한 암세포 사멸 증진 및 전이 억제 효과, 주화성 억제 효과, 및 항 천식 효과 등을 실험적으로 확인하였는바, 염증성 질환을 치료하기 위한 약학적 조성물로 유용하게 사용될 수 있을 것으로 기대된다.The present invention relates to a novel compound that exhibits Leukotriene B4 receptor 2 (BLT2) inhibitory activity and a pharmaceutical composition for preventing or treating an inflammatory disease comprising the same. The present inventors have identified a novel compound that exhibits BTL2 inhibitory activity in order to solve in vivo instability and difficulty in mass production, which are problems of conventional inflammatory disease treatment substances, and has excellent cancer cell death enhancement and metastasis suppression effect and chemotaxis inhibition of the compound. The effects, and anti-asthma effects have been confirmed experimentally, it is expected that it can be usefully used as a pharmaceutical composition for treating inflammatory diseases.
도 1a 내지 도 1e는 BLT2가 발현된 세포 (CHO-BLT2)에서, 본 발명의 화합물 처리에 의한 성장 억제 효과를 확인한 결과이다.1A to 1E show the results of confirming the growth inhibitory effect of the compound of the present invention in cells expressing BLT2 (CHO-BLT2).
도 2a 및 도 2b는 BLT2가 발현된 세포 (CHO-BLT2 cells)에서, 본 발명의 화합물 처리에 의한 세포의 주화성 억제 효과 및 IC50 (50% 억제 농도)를 확인한 결과이다. Figures 2a and 2b is a result of confirming the chemotaxis inhibitory effect and IC 50 (50% inhibitory concentration) of the cells by the compound treatment of the present invention in CLT-expressing cells (CHO-BLT2 cells).
도 3a 및 도 3b는 BLT2가 발현된 세포 (CHO-BLT2 cells) 또는 BLT1이 발현된 세포 (CHO-BLT1)에서, 본 발명의 화합물 처리에 의한 세포의 주화성 억제 효과를 확인한 결과이다.3A and 3B show the results of confirming the chemotaxis inhibitory effect of cells treated with the compound of the present invention in cells expressing BLT2 (CHO-BLT2 cells) or cells expressing BLT1 (CHO-BLT1).
도 4a 및 도 4b는 BLT2가 발현된 세포 (CHO-BLT2 cells)에서, 본 발명의 화합물 처리에 의한 LTB4와 BLT2 결합 저해 효과를 확인한 결과이다.Figures 4a and 4b is a result of confirming the effect of inhibiting the binding of LTB 4 and BLT2 by the compound treatment of the present invention in BLT2-expressing cells (CHO-BLT2 cells).
도 5a 및 도 5b는 MDA-MB-231 세포 또는 MDA-MB-435 세포에서, 본 발명의 화합물 처리에 의한 활성 산소의 생성 억제효과를 확인한 결과이다. Figures 5a and 5b is the result of confirming the inhibitory effect of the production of active oxygen by the compound treatment of the present invention in MDA-MB-231 cells or MDA-MB-435 cells.
도 6a 및 도 6b는 MDA-MB-231 세포 또는 MDA-MB-435 세포에서, 본 발명의 화합물 처리에 의한 IL-8 (interleukin-8) 발현량 억제효과를 확인한 결과이다. Figures 6a and 6b is a result of confirming the inhibitory effect of IL-8 (interleukin-8) expression by the compound treatment of the present invention in MDA-MB-231 cells or MDA-MB-435 cells.
도 7a 및 도 7b는 MDA-MB-231 세포 또는 MDA-MB-435 세포에서, 본 발명의 화합물 처리에 의한 암세포 침투 억제효과를 확인한 결과이다.Figures 7a and 7b is a result of confirming the cancer cell infiltration inhibition effect by the compound treatment of the present invention in MDA-MB-231 cells or MDA-MB-435 cells.
도 8, 도 9a 및 도 9b는 본 발명의 화합물 처리에 의한 암세포 전이 억제효과를 확인한 결과이다. 8, 9a and 9b is a result confirming the cancer cell metastasis inhibitory effect by the compound treatment of the present invention.
도 10은 중증 천식이 유도된 마우스에서, 본 발명의 화합물 처리에 의한 기도 과민성 감소 효과를 확인한 결과이다.10 is a result of confirming the effect of reducing airway hypersensitivity by the compound treatment of the present invention in mice with severe asthma induced.
도 11은 중증 천식이 유도된 마우스에서, 본 발명의 화합물 처리에 의한 IL-4 (interleukin-4) 생성 감소 효과를 확인한 결과이다.11 is a result confirming the effect of reducing IL-4 (interleukin-4) production by the compound treatment of the present invention in mice with severe asthma induced.
도 12는 천식이 유도된 마우스에서, 본 발명의 화합물 처리에 의한 기도 과민성 감소 효과를 확인한 결과이다.12 is a result confirming the effect of reducing airway hypersensitivity by the compound treatment of the present invention in asthma induced mice.
도 13a 내지 도 13c는 천식이 유도된 마우스에서, 본 발명의 화합물 처리에 의한 마우스 복강에서 유입된 전체 세포와 호중구가 감소됨을 확인한 결과이다.13A to 13C show that asthma-induced mice reduced total cells and neutrophils introduced into the abdominal cavity of the mouse by the compound treatment of the present invention.
도 14a 및 도 14b는 천식이 유도된 마우스에서, 본 발명의 화합물 처리에 의한 마우스 복강에서 유입된 전체 세포와 호중구가 감소됨을 확인한 결과이다.14A and 14B are results confirming that the total cells and neutrophils introduced into the abdominal cavity of the mouse by the compound treatment of the present invention are reduced in asthma-induced mice.
본 발명자들은, 실시예에서 제조한 신규 화합물을 처리한 경우, BLT2 발현 세포의 성장을 현저히 억제시킬 수 있다는 점에 기반하여 상기 화합물의 암세포 사멸 증진, 암세포 전이 억제, BLT2 의존적인 주화성 저해 및 항천식 효과 등을 구체적으로 확인하고, 이에 기초하여 본 발명을 완성하였다.The present inventors, when treated with the novel compound prepared in Example, can significantly inhibit the growth of BLT2 expressing cells, thereby promoting cancer cell death, inhibiting cancer cell metastasis, inhibiting BLT2-dependent chemotaxis and The asthma effect, etc. were specifically confirmed and the present invention was completed based on this.
이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 하기 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용 가능한 염을 제공한다.The present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.
[화학식 1][Formula 1]
Figure PCTKR2016008069-appb-I000001
Figure PCTKR2016008069-appb-I000001
상기 화학식 1에서, In Chemical Formula 1,
R1은 C1~C10의 알킬이고,R 1 is C 1 -C 10 is alkyl,
R2는
Figure PCTKR2016008069-appb-I000002
,
Figure PCTKR2016008069-appb-I000003
,
Figure PCTKR2016008069-appb-I000004
,
Figure PCTKR2016008069-appb-I000005
또는
Figure PCTKR2016008069-appb-I000006
이며, R 2 is
Figure PCTKR2016008069-appb-I000002
,
Figure PCTKR2016008069-appb-I000003
,
Figure PCTKR2016008069-appb-I000004
,
Figure PCTKR2016008069-appb-I000005
or
Figure PCTKR2016008069-appb-I000006
Is,
Ra는
Figure PCTKR2016008069-appb-I000007
,
Figure PCTKR2016008069-appb-I000008
,
Figure PCTKR2016008069-appb-I000009
,
Figure PCTKR2016008069-appb-I000010
,
Figure PCTKR2016008069-appb-I000011
,
Figure PCTKR2016008069-appb-I000012
,
Figure PCTKR2016008069-appb-I000013
,
Figure PCTKR2016008069-appb-I000014
,
Figure PCTKR2016008069-appb-I000015
,
Figure PCTKR2016008069-appb-I000016
,
Figure PCTKR2016008069-appb-I000017
,
Figure PCTKR2016008069-appb-I000018
,
Figure PCTKR2016008069-appb-I000019
,
Figure PCTKR2016008069-appb-I000020
,
Figure PCTKR2016008069-appb-I000021
,
Figure PCTKR2016008069-appb-I000022
,
Figure PCTKR2016008069-appb-I000023
,
Figure PCTKR2016008069-appb-I000024
,
Figure PCTKR2016008069-appb-I000025
,
Figure PCTKR2016008069-appb-I000026
,
Figure PCTKR2016008069-appb-I000027
,
Figure PCTKR2016008069-appb-I000028
또는 하이드록시이고,R a is
Figure PCTKR2016008069-appb-I000007
,
Figure PCTKR2016008069-appb-I000008
,
Figure PCTKR2016008069-appb-I000009
,
Figure PCTKR2016008069-appb-I000010
,
Figure PCTKR2016008069-appb-I000011
,
Figure PCTKR2016008069-appb-I000012
,
Figure PCTKR2016008069-appb-I000013
,
Figure PCTKR2016008069-appb-I000014
,
Figure PCTKR2016008069-appb-I000015
,
Figure PCTKR2016008069-appb-I000016
,
Figure PCTKR2016008069-appb-I000017
,
Figure PCTKR2016008069-appb-I000018
,
Figure PCTKR2016008069-appb-I000019
,
Figure PCTKR2016008069-appb-I000020
,
Figure PCTKR2016008069-appb-I000021
,
Figure PCTKR2016008069-appb-I000022
,
Figure PCTKR2016008069-appb-I000023
,
Figure PCTKR2016008069-appb-I000024
,
Figure PCTKR2016008069-appb-I000025
,
Figure PCTKR2016008069-appb-I000026
,
Figure PCTKR2016008069-appb-I000027
,
Figure PCTKR2016008069-appb-I000028
Or hydroxy,
Rb는
Figure PCTKR2016008069-appb-I000029
,
Figure PCTKR2016008069-appb-I000030
,
Figure PCTKR2016008069-appb-I000031
또는
Figure PCTKR2016008069-appb-I000032
이고,R b is
Figure PCTKR2016008069-appb-I000029
,
Figure PCTKR2016008069-appb-I000030
,
Figure PCTKR2016008069-appb-I000031
or
Figure PCTKR2016008069-appb-I000032
ego,
RC는
Figure PCTKR2016008069-appb-I000033
,
Figure PCTKR2016008069-appb-I000034
또는
Figure PCTKR2016008069-appb-I000035
이고, R C is
Figure PCTKR2016008069-appb-I000033
,
Figure PCTKR2016008069-appb-I000034
or
Figure PCTKR2016008069-appb-I000035
ego,
Rd는 수소 또는
Figure PCTKR2016008069-appb-I000036
이고,R d is hydrogen or
Figure PCTKR2016008069-appb-I000036
ego,
Re는
Figure PCTKR2016008069-appb-I000037
또는
Figure PCTKR2016008069-appb-I000038
이고,R e is
Figure PCTKR2016008069-appb-I000037
or
Figure PCTKR2016008069-appb-I000038
ego,
R3는 수소 또는 플루오르일 수 있다.R 3 may be hydrogen or fluorine.
본 발명에 따른 상기 화학식 1로 표시되는 화합물의 바람직한 예는 하기와 같다:Preferred examples of the compound represented by Formula 1 according to the present invention are as follows:
tert-부틸 4-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-(4-(4-메틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-에틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-(2-하이드록시에틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-(사이클로프로필메틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-사이클로헥실피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-(사이클로헥실메틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(4-(4-아이소부틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-페닐-N-(3-(4-(4-(프로프-2-이닐)피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-(4-(4-시아노피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; tert -부틸 4-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; N-(3-플루오로페닐)-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-플루오로페닐)-N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; tert-부틸 4-(4-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; N-(4-플루오로페닐)-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(4-플루오로페닐)-N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-(4-(몰폴린-4-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-페닐-N-(3-(4-(피페리딘-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N,N-다이에틸-4-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤즈아마이드; N-페닐-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-(3-(4-메틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N-(3-(3-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; tert-부틸-4-(3-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; N-(4-플루오로페닐)-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(4-플루오로페닐)-N-(3-(3-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N-(3-(4-하이드록시페닐)프로프-2-이닐)-N-페닐펜탄아마이드; 2-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)페녹시)아세트산; tert-부틸 4-(5-(3-((N-페닐펜탄아미도)프로프-1-인-1-일)피콜리노일)피페라진-1-카복실레이트; N-페닐-N-(3-(6-(피페라진-1-카보닐)피리딘-3-일)프로프-2-인-1-일)펜탄아마이드; N-(3-(6-아이소프로필피페라진-1-카보닐)피리딘-3-일)프로프-2-인-1일)펜탄아마이드; N,N-다이에틸-4-(3-(N-(3-플루오로페닐)펜타아미도)프로프-1-인-1-일)벤즈아마이드; N,N-다이에틸-4-(3-(N-(4-플루오로페닐)펜타아미도)프로프-1-인-1-일)벤즈아마이드; N-(3-(4-(N,N-다이에틸설파모일)페닐)프로프-2-이닐)-N-페닐펜타아마이드; N-(3-(4-(N-아이소프로필설파모일)페닐)프로프-2-이닐)-N-페닐펜타아마이드; tert-부틸 4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤조에이트; 4-(3-(N-페닐펜탄아미도)프로-1-핀-1-일)벤조익산; N-에틸-4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아마이드; N-(2-(다이에틸아미노)에틸)-4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아마이드; 에틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)아세테이트; 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)아세틱산; 메틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)프로파노에이트; 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)프로피오닉산; 2-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세트산; 및 2-(4-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세트산. tert -butyl 4- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) benzoyl) piperazin-1-carboxylate; N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide; N- (3- (4- (4-methylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N- (3- (4- (4-ethylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N- (3- (4- (4-isopropylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N- (3- (4- (4- (2-hydroxyethyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N- (3- (4- (4- (cyclopropylmethyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N- (3- (4- (4-cyclohexylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N- (3- (4- (4- (cyclohexylmethyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N- (3- (4- (4-isobutylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; N -phenyl- N- (3- (4- (4- (prop-2-ynyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide; N- (3- (4- (4-cyanopiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; tert - butyl 4- (4- (3- ( N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate; N - (3-fluorophenyl) - N - (3- (4- ( piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; N - (3-fluorophenyl) - N - (3- (4- (4- isopropyl-piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; tert -butyl 4- (4- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate; N - (4-fluorophenyl) - N - (3- (4- ( piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; N - (4-fluorophenyl) - N - (3- (4- (4-isopropyl-piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; N- (3- (4- (morpholin-4-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N -phenyl- N- (3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide; N , N -diethyl-4- (3- ( N -phenylpentaneamido) prop-1-ynyl) benzamide; N -phenyl- N- (3- (3- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide; N- (3- (3- (4-methylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide; N- (3- (3- (4-isopropylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide; tert -butyl-4- (3- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate; N - (4-fluorophenyl) - N - (3- (3- ( piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; N - (4- fluorophenyl) - N - (3- (3- (4- isopropyl-piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide; N- (3- (4-hydroxyphenyl) prop-2-ynyl) -N -phenylpentanamide; 2- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) phenoxy) acetic acid; tert -butyl 4- (5- (3-(( N -phenylpentaneamido) prop-1-yn-1-yl) picolinoyl) piperazin-1-carboxylate; N -phenyl- N- ( 3- (6- (piperazin-1-carbonyl) pyridin-3-yl) prop-2-yn-1-yl) pentaneamide; N- (3- (6-isopropylpiperazin-1-carbo Yl) pyridin-3-yl) prop-2-yn-1 yl) pentaneamide; N, N -diethyl-4- (3- ( N- (3-fluorophenyl) pentamido) prop-1-yn-1-yl) benzamide; N, N -diethyl-4- (3- ( N- (4-fluorophenyl) pentamido) prop-1-yn-1-yl) benzamide; N- (3- (4- ( N, N -diethylsulfamoyl) phenyl) prop-2-ynyl) -N -phenylpentamide; N- (3- (4- ( N -isopropylsulfamoyl) phenyl) prop-2-ynyl) -N -phenylpentamide; tert -butyl 4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzoate; 4- (3- ( N -phenylpentanamido) pro-1-pin-1-yl) benzoic acid; N -ethyl-4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamide; N- (2- (diethylamino) ethyl) -4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamide; Ethyl 2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetate; 2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetic acid; Methyl 2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) propanoate; 2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) propionic acid; 2- (4- (3- ( N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy) acetic acid; And 2- (4- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy) acetic acid.
본 발명에서 사용되는 "약학적으로 허용되는"이라는 용어는 과도한 독성, 자극, 알러지 반응 또는 기타 문제점 또는 합병증 없이 이득/위험 비가 합리적이어서 대상체 (예: 인간)의 조직과 접촉하여 사용하기에 적합하며 건전한 의학적 판단의 범주 이내인 화합물 또는 조성물을 의미한다.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.
본 발명에서 사용되는 용어 "염"은 약학적으로 허용 가능한 유리산(free acid)에 의해 형성된 산 부가염이 유용하다. 산 부가염은 염산, 질산, 인산, 황산, 브롬화수소산, 요드화수소산, 아질산 또는 아인산과 같은 무기산류와 지방족 모노 및 디카르복실레이트, 페닐-치환된 알카노에이트, 하이드록시 알카노에이트 및 알칸디오에이트, 방향족 산류, 지방족 및 방향족 설폰산류와 같은 무독성 유기산으로부터 얻는다. 이러한 약학적으로 무독한 염류로는 설페이트, 피로설페이트, 바이설페이트, 설파이트, 바이설파이트, 니트레이트, 포스페이트, 모노하이드로겐 포스페이트, 디하이드로겐 포스페이트, 메타포스페이트, 피로포스페이트 클로라이드, 브로마이드, 아이오다이드, 플루오라이드, 아세테이트, 프로피오네이트, 데카노에이트, 카프릴레이트, 아크릴레이트, 포메이트, 이소부티레이트, 카프레이트, 헵타노에이트, 프로피올레이트, 옥살레이트, 말로네이트, 석시네이트, 수베레이트, 세바케이트, 푸마레이트, 말리에이트, 부틴-1,4-디오에이트, 헥산-1,6-디오에이트, 벤조에이트, 클로로벤조에이트, 메틸벤조에이트, 디니트로 벤조에이트, 하이드록시벤조에이트, 메톡시벤조에이트, 프탈레이트, 테레프탈레이트, 벤젠설포네이트, 톨루엔설포네이트, 클로로벤젠설포네이트, 크실렌설포네이트, 페닐아세테이트, 페닐프로피오네이트, 페닐부티레이트, 시트레이트, 락테이트, β-하이드록시부티레이트, 글리콜레이트, 말레이트, 타트레이트, 메탄설포네이트, 프로판설포네이트, 나프탈렌-1-설포네이트, 나프탈렌-2-설포네이트 또는 만델레이트를 포함한다.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.
본 발명에 따른 산 부가염은 통상의 방법, 예를 들면, 화학식 1 내지 4로 표시되는 화합물을 과량의 산 수용액 중에 용해시키고, 이 염을 수혼화성 유기 용매, 예를 들면 메탄올, 에탄올, 아세톤 또는 아세토니트릴을 사용하여 침전시켜서 제조할 수 있다. 또한 이 혼합물에서 용매나 과량의 산을 증발시킨 후 건조시키거나 또는 석출된 염을 흡입 여과시켜 제조할 수도 있다.Acid addition salts according to the invention are dissolved in conventional methods, for example, the compounds represented by the formulas (1) to (4) in an excess of aqueous acid solution, and the salts are water miscible organic solvents 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.
또한, 염기를 사용하여 약학적으로 허용 가능한 금속염을 만들 수도 있다. 알칼리 금속 또는 알칼리 토금속 염은 예를 들면, 화합물을 과량의 알칼리 금속 수산화물 또는 알칼리 토금속 수산화물 용액 중에 용해하고, 비용해 화합물 염을 여과하고, 여액을 증발, 건조시켜 얻는다. 이때, 금속염으로는 나트륨, 칼륨 또는 칼슘염을 제조하는 것이 제약상 적합하다. 이에 대응하는 은염은 알칼리 금속 또는 알칼리 토금속 염을 적당한 음염 (예, 질산은)과 반응시켜 얻는다.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).
본 발명의 일실시예에서는 BLT2 억제 활성을 나타내는 신규 화합물을 제조하였으며 (실시예 1 내지 46 참조), 상기 신규 화합물 처리에 의한 BLT2 발현 세포의 성장 억제를 확인하였다 (실험예 2 참조). 또한, BLT2 발현 세포의 주화성을 억제시킬 수 있음을 확인하였다 (실험예 3 참조). 또한, 상기 화합물을 이용하여 LTB4와 BLT2 결합 저해효과를 확인하였고 (실험예 4 참조), 세포 내 활성 산소 억제, IL-8 발현 억제, 암세포 침투 억제 및 암세포의 전이 억제를 확인하였으며 (실험예 5 참조), 천식이 유도된 마우스에서, 기도 과민성 감소, IL-4 생성 억제 및 마우스 복강으로의 면역세포 유입 억제 효과를 구체적으로 확인하였는바 (실험예 6 참조) 염증성 질환 약학적 조성물로 매우 유용하게 사용될 수 있음을 확인하였다.In one embodiment of the present invention was prepared a novel compound exhibiting BLT2 inhibitory activity (see Examples 1 to 46), and confirmed the growth inhibition of BLT2 expressing cells by the novel compound treatment (see Experimental Example 2). In addition, it was confirmed that chemotaxis of BLT2 expressing cells can be suppressed (see Experimental Example 3). In addition, the compound was used to confirm the inhibitory effect of LTB 4 and BLT2 binding (see Experimental Example 4), inhibited intracellular free radicals, inhibited IL-8 expression, inhibited cancer cell infiltration and inhibited metastasis of cancer cells (Experimental Example). 5), the effect of reducing airway hypersensitivity, inhibiting IL-4 production, and inhibiting immune cell influx into the mouse abdominal cavity was specifically confirmed in asthma-induced mice (see Experimental Example 6). It was confirmed that it can be used.
이에, 본 발명은 상기 화합물 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는, 염증성 질환 예방 또는 치료용 약학적 조성물을 제공한다.Accordingly, the present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease, comprising the compound or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명에서 사용되는 용어, "예방"이란 본 발명에 따른 약학적 조성물의 투여에 의해 염증성 질환을 억제시키거나 발병을 지연시키는 모든 행위를 의미한다.As used herein, the term "prevention" means any action that inhibits or delays the development of an inflammatory disease by administration of a pharmaceutical composition according to the present invention.
본 발명에서 사용되는 용어, "치료"란 본 발명에 따른 약학적 조성물의 투여에 의해 염증성 질환에 대한 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미한다.As used herein, the term "treatment" means any action that improves or advantageously alters the symptoms of an inflammatory disease by administration of a pharmaceutical composition according to the present invention.
본 발명에서, 염증성 질환은 BLT2 (Leukotriene B4 receptor 2)의 과발현에 기인한 질병으로, 천식, 죽상경화증, 암, 피부가려움증, 류마티스 관절염 및 염증성 장 질환에서 선택되는 1종 이상일 수 있으나, 이로써 제한되는 것은 아니다. 본 명세서에서 예시한 상기 질환 외에도, 당업계에 알려져 있는 BLT2-연관된 염증성 질환은 모두 본 발명의 화학식 1의 구조를 갖는 화합물로 예방 또는 치료할 수 있는 염증성 질환에 포함되는 것으로 본다. 한 구체적인 예에서, 상기 암은 BLT2 또는 종양 유전자 Ras의 과발현에 의해 유발된 임의의 암일 수 있다. 이에 제한되는 것은 아니나, 상기 암은 방광암, 전립선암, 췌장암, 유방암, 뇌종양, 피부암 및 간암으로 이루어진 군으로부터 선택되는 것일 수 있으며, 이로써 제한되는 것은 아니다. In the present invention, the inflammatory disease is due to overexpression of Leukotriene B4 receptor 2 (BLT2), and may be at least one selected from asthma, atherosclerosis, cancer, skin itch, rheumatoid arthritis and inflammatory bowel disease, but is limited thereto. It is not. In addition to the diseases exemplified herein, all BLT2-associated inflammatory diseases known in the art are considered to be included in inflammatory diseases that can be prevented or treated with a compound having the structure of Formula 1 of the present invention. In one specific example, the cancer may be any cancer caused by overexpression of BLT2 or the oncogene Ras. Although not limited thereto, the cancer may be selected from the group consisting of bladder cancer, prostate cancer, pancreatic cancer, breast cancer, brain tumor, skin cancer, and liver cancer, but is not limited thereto.
본 발명에서, BLT2 (Leukotriene B4 receptor 2)는 GPCR(G protein-coupled receptor) 군 중 하나로 LTB4 (Leukotriene B4; LTB4)에 대해 낮은 친화력을 갖는 수용체로서, 본 발명의 조성물은 BLT2에 의한 세포 성장을 억제함으로써, 염증성 질환을 예방 또는 치료할 수 있다. 보다 구체적으로 BLT2 활성으로 유도된 ROS의 생성을 저해하여 LTB4-유도된 주화성을 저해할 수 있다.In the present invention, BLT2 (Leukotriene B4 receptor 2) LTB 4 is one of the group GPCR (G protein-coupled receptor) As a receptor having low affinity for (Leukotriene B4; LTB 4 ), the composition of the present invention can prevent or treat inflammatory diseases by inhibiting cell growth by BLT2. More specifically, the inhibition of the production of ROS induced by BLT2 activity may inhibit LTB 4 -induced chemotaxis.
본 발명에서 사용되는 용어, "저해"는 유전자의 전사, mRNA 프로세싱, 번역, 전좌 및 성숙 중 임의의 단계를 저해하거나, 단백질과 단백질간의 결합, 단백질의 활성화 또는 이를 통한 신호전달의 저해를 의미한다.As used herein, the term "inhibition" means inhibiting any step of transcription, mRNA processing, translation, translocation, and maturation of a gene, or inhibition of protein-to-protein binding, activation of a protein, or signaling through it. .
본 발명의 약학적 조성물은 유효성분 이외에 약제학적으로 허용되는 담체를 포함할 수 있다. 이때, 약제학적으로 허용되는 담체는 제제 시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세 결정성셀룰로스, 폴리비닐피로리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필 히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일등을 포함하나, 이에 한정되는 것은 아니다. 또한, 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다.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.
본 발명의 약학적 조성물은 목적하는 방법에 따라 경구 투여하거나 비경구투여(예를 들어, 정맥 내, 피하, 복강 내 또는 국소에 적용)할 수 있으며, 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 시간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다.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.
본 발명의 약학적 조성물은 약학적으로 유효한 양으로 투여한다. 본 발명에 있어서 "약학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효용량 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명에 다른 약학적 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.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 single or multiple administrations. 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.
구체적으로 본 발명의 약학적 조성물의 유효량은 환자의 연령, 성별, 상태, 체중, 체내에 활성 성분의 흡수도, 불활성율 및 배설속도, 질병종류, 병용되는 약물에 따라 달라질 수 있으며, 일반적으로는 체중 1kg 당 0.001 내지 150mg, 바람직하게는 0.01내지 100mg을 매일 또는 격일 투여하거나, 1일 1 내지 3회로 나누어 투여할 수 있다. 그러나 투여 경로, 비만의 중증도, 성별, 체중, 연령 등에 따라서 증감 될 수 있으므로 상기 투여량이 어떠한 방법으로도 본 발명의 범위를 한정하는 것은 아니다.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.
또한, 본 발명은 상기 약학적 조성물을 개체에 투여하는 단계를 포함하는 염증성 질환의 치료방법을 제공한다. 본 발명에서 "개체"란 질병의 치료를 필요로 하는 대상을 의미하고, 보다 구체적으로는, 인간 또는 비-인간인 영장류, 생쥐(mouse), 개, 고양이, 말 및 소 등의 포유류를 의미한다.The present invention also provides a method of treating an inflammatory 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. .
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시한다. 그러나 하기의 실시예는 본 발명을 보다 쉽게 이해하기 위하여 제공되는 것일 뿐, 하기 실시예에 의해 본 발명의 내용이 한정되는 것은 아니다.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.
[실시예]EXAMPLE
실시예Example 1.  One. terttert -부틸 4-(4-(3-(-Butyl 4- (4- (3- ( NN -- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1--One- 이닐Inil )) 벤조일Benzoyl )피페라진-1-카복실레이트 (Piperazine-1-carboxylate ( terttert -butyl 4-(4-(3-(-butyl 4- (4- (3- ( NN -- phenylpentanamidophenylpentanamido )prop-1-ynyl)benzoyl)piperazine-1-carboxylate) (LMT-693)의 제조) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) (LMT-693)
단계 1 : Step 1: NN -페닐펜탄아마이드(-Phenylpentaneamide ( NN -phenylpentanamide)의 제조-phenylpentanamide)
아닐린 (aniline)(0.98 ml, 10.74 mmol)을 다이클로로메탄(20 ml)에 녹인 후 얼음에서 냉각시켰다. 상기 혼합물에 트리에틸아민 (3.00 ml, 21.48 mmol)을 가한 뒤 5분 동안 교반하였다. 같은 온도에서 발레로일 클로라이드 (valeroyl chloride)(2.60 ml, 21.48 mmol)를 가하고, 얼음을 제거한 뒤, 상기 혼합물을 상온에서 2시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘 (MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피 (Hex:EA=10:1)로 정제하여 N-페닐펜탄아마이드 (N-phenylpentanamide)를 수득하였다 (1.88 g, 99% 수율).Aniline (0.98 ml, 10.74 mmol) was dissolved in dichloromethane (20 ml) and cooled on ice. Triethylamine (3.00 ml, 21.48 mmol) was added to the mixture and stirred for 5 minutes. Valeroyl chloride (2.60 ml, 21.48 mmol) was added at the same temperature, ice was removed, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 10: 1 ) to give N - pentane to give the phenyl-amide (N -phenylpentanamide) (1.88 g, 99% yield).
단계 2 : Step 2: terttert -부틸 4-(4--Butyl 4- (4- 브로모벤조일Bromobenzoyl )피페라진-1-Piperazine-1- 카복실레이트(Carboxylate ( terttert -butyl -butyl 4-(4-bromobenzoyl)piperazine-1-carboxylate)의 제조Preparation of 4- (4-bromobenzoyl) piperazine-1-carboxylate)
4-브로모벤조산 (4-bromobenzoic acid)(901 mg, 4.48 mmol)과 tert-부틸 피페라진-1-카복실레이트 (tert-butyl piperazine-1-carboxylate)(1.00 g, 5.37 mmol)를 N,N-다이메틸포름아마이드 (DMF)(15 ml)에 녹인 후, 5분 동안 교반하였다. 상기 혼합물에 HATU(1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate)(2.04 g, 5.37 mmol)와 N,N-다이아이소프로필에틸아민(DIPEA)(2.34 ml, 13.44 mmol)을 가하고, 상온에서 15시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘 (MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=2:1)로 정제하여 tert-부틸 4-(4-브로모벤조일)피페라진-1-카복실레이트(tert-butyl 4-(4-bromobenzoyl)piperazine-1-carboxylate)를 수득하였다 (1.56 g, 94% 수율).4-bromo-benzoic acid (4-bromobenzoic acid) (901 mg, 4.48 mmol) and tert - butyl piperazine-1-carboxylate (tert -butyl piperazine-1-carboxylate ) (1.00 g, 5.37 mmol) N, N Dissolve in dimethylformamide (DMF) (15 ml) and stir for 5 minutes. HATU (1- [Bis (dimethylamino) methylene] -1H-1,2,3-triazolo [4,5-b] pyridinium 3-oxid hexafluorophosphate) (2.04 g, 5.37 mmol) and N , N -diamond were added to the mixture. Isopropylethylamine (DIPEA) (2.34 ml, 13.44 mmol) was added and stirred at room temperature for 15 hours. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 2: 1) and tert -butyl 4- (4-bromobenzoyl) piperazine-1-carboxylate ( tert- butyl 4- (4-bromobenzoyl) piperazine -1-carboxylate) was obtained (1.56 g, 94% yield).
단계 3 : Step 3: terttert -부틸 4-(4-(3--Butyl 4- (4- (3- 하이드록시프로프Hydroxyprop -1--One- 이닐Inil )) 벤조일Benzoyl )피페라진-1-Piperazine-1- 카복실레이트Carboxylate (( terttert -butyl 4-(4-(3--butyl 4- (4- (3- hydroxyprophydroxyprop -1--One- ynylynyl )) benzoylbenzoyl )) piperazinepiperazine -1-carboxylate)의 제조-1-carboxylate)
상기 단계 2에서 수득한 tert-부틸 4-(4-브로모벤조일)피페라진-1-카복실레이트(1.00 g, 2.71 mmol)와 프로파길 알코올 (propagyl alcohol)(0.32 ml, 5.42 mmol)을 트리에틸아민 (12 ml)에 녹인 후, 5분 동안 교반하였다. 상기 혼합물에 비스(트리페닐포스핀)팔라듐(II) 다이클로라이드 (190 mg, 0.271 mmol)와 요오드화 구리 (I)(52 mg, 0.271 mmol)를 가하고 60 ℃에서 가열, 환류하여 17시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 감압 하에 농축한 뒤, 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘 (MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피 (Hex:EA=2:1)로 정제하여 tert-부틸 4-(4-(3-하이드록시프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (tert-butyl 4-(4-(3-hydroxyprop-1-ynyl)benzoyl)piperazine-1-carboxylate)를 수득하였다 (850 mg, 91% 수율). Triethyl tert -butyl 4- (4-bromobenzoyl) piperazine-1-carboxylate (1.00 g, 2.71 mmol) and propagyl alcohol (0.32 ml, 5.42 mmol) obtained in step 2 were obtained. Dissolved in amine (12 ml) and stirred for 5 minutes. Bis (triphenylphosphine) palladium (II) dichloride (190 mg, 0.271 mmol) and copper iodide (I) (52 mg, 0.271 mmol) were added to the mixture, which was heated and refluxed at 60 ° C. for 17 hours. . The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 2: 1) to give tert -butyl 4- (4- (3-hydroxyprop-1-ynyl) benzoyl) piperazine-1-carboxylate ( tert -butyl 4- (4- (3-hydroxyprop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (850 mg, 91% yield).
단계 4 : Step 4: terttert -부틸 4-(4-(3-(-Butyl 4- (4- (3- ( 메틸설포닐옥시Methylsulfonyloxy )) 프로프Prof -1--One- 이닐Inil )) 벤조일Benzoyl )피페라진-1-카복실레이트(Piperazine-1-carboxylate ( terttert -butyl 4-(4-(3-(-butyl 4- (4- (3- ( methylsulfonyloxymethylsulfonyloxy )prop-1-ynyl)benzoyl)piperazine-1-carboxylate)의 제조) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
상기 단계 3에서 수득한 tert-부틸 4-(4-(3-하이드록시프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (600 mg, 1.74 mmol)를 다이클로로메탄 (8 ml)에 녹인 후 얼음에서 냉각시켰다. 상기 혼합물에 트리에틸아민(0.36 ml, 2.61 mmol)을 가한 뒤 5분 동안 교반하였다. 같은 온도에서 메탄설포닐 클로라이드(0.15 ml, 1.92 mmol)를 가하고, 얼음을 제거한 뒤, 상기 혼합물을 상온에서 30분 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산 마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=2:1)로 정제하여 tert-부틸 4-(4-(3-(메틸설포닐옥시)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (tert-butyl 4-(4-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylate)를 수득하였다(662 mg, 90% 수율). Tert -butyl 4- (4- (3-hydroxyprop-1-ynyl) benzoyl) piperazine-1-carboxylate (600 mg, 1.74 mmol) obtained in step 3 was diluted with dichloromethane (8 ml). It was dissolved in and cooled on ice. Triethylamine (0.36 ml, 2.61 mmol) was added to the mixture, which was then stirred for 5 minutes. Methanesulfonyl chloride (0.15 ml, 1.92 mmol) was added at the same temperature, ice was removed, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 2: 1) to give tert -butyl 4- (4- (3- (methylsulfonyloxy) prop-1-ynyl) benzoyl) piperazin-1- A carboxylate ( tert -butyl 4- (4- (3- (methylsulfonyloxy) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (662 mg, 90% yield).
단계 5 : Step 5: terttert -- 부틸4Butyl 4 -(4-(3-(-(4- (3- ( NN -- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1--One- 이닐Inil )) 벤조일Benzoyl )피페라진-1-카복실레이트(Piperazine-1-carboxylate ( terttert -butyl 4-(4-(3-(-butyl 4- (4- (3- ( NN -- phenylpentanamidophenylpentanamido )prop-1-ynyl)benzoyl)piperazine-1-carboxylate)의 제조) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
상기 단계 1에서 수득한 N-페닐펜탄아마이드 (185 mg, 1.04 mmol)와 나트륨 하이드라이드 (NaH)(75 mg, 3.12 mmol)를 얼음에서 냉각시킨 후, 테트라하이드로퓨란 (THF)(8 ml)을 넣고 30분 동안 교반하였다. 상기 혼합물에 상기 단계 4에서 수득한 tert-부틸 4-(4-(3-(메틸설포닐옥시)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (662 mg, 1.57 mmol)를 가하고, 얼음을 제거한 뒤, 상기 혼합물을 상온에서 17시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘 (MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피 (Hex:EA=4:1)로 정제하여 최종 생성물인 tert-부틸 4-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (tert-butyl 4-(4-(3-(N-phenylpentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate)를 수득하였다(382 mg, 73% 수율).After cooling N -phenylpentanamide (185 mg, 1.04 mmol) and sodium hydride (NaH) (75 mg, 3.12 mmol) obtained in step 1 on ice, tetrahydrofuran (THF) (8 ml) was added thereto. Put and stirred for 30 minutes. To the mixture was added tert -butyl 4- (4- (3- (methylsulfonyloxy) prop-1-ynyl) benzoyl) piperazine-1-carboxylate (662 mg, 1.57 mmol) obtained in step 4. After the addition of ice, the mixture was stirred at room temperature for 17 hours. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 4: 1) to give tert -butyl 4- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) benzoyl) as a final product. Piperazine-1-carboxylate ( tert- butyl 4- (4- (3- ( N- phenylpentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (382 mg, 73% yield).
1H NMR (CDCl3, 500MHz) δ 7.40-7.20(9H, m), 4.65(2H, s), 3.62-3.32(8H, br), 2.02-1.97(2H, t), 1.52-1.48(2H, m), 1.40(9H, s), 1.19-1.12(2H, m), 0.76-0.72(3H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.40-7.20 (9H, m), 4.65 (2H, s), 3.62-3.32 (8H, br), 2.02-1.97 (2H, t), 1.52-1.48 (2H, m), 1.40 (9H, s), 1.19-1.12 (2H, m), 0.76-0.72 (3H, t).
실시예Example 2.  2. NN -페닐--Phenyl- NN -(3-(4-(피페라진-1--(3- (4- (piperazin-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )) 펜탄아마이드Pentaneamide ( ( NN -phenyl--phenyl- NN -(3-(4-(-(3- (4- ( piperazinepiperazine -1-carbonyl)phenyl)prop-2--1-carbonyl) phenyl) prop-2- ynylynyl )) pentanamidepentanamide ) (LMT-694)의 제조) (LMT-694) Preparation
상기 실시예 1에서 수득한 tert-부틸 4-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (754 mg, 1.50 mmol)를 아세토나이트릴 (15 ml)에 녹인 후, 상온에서 5분 동안 교반하였다. 상기 혼합물에 다이옥산 (dioxane)과 혼합한 하이드로클로라이드 (4N)(3.73 ml)를 가하고, 같은 온도에서 1시간 30분 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 실리카겔 컬럼 크로마토그래피 (CH2Cl2:MeOH=50:1)로 정제하여 최종 생성물인 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드(N-phenyl-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide)를 수득하였다 (363 mg, 60% 수율). Tert -butyl 4- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate (754 mg, 1.50 mmol) obtained in Example 1 was prepared. After dissolving in acetonitrile (15 ml), it was stirred for 5 minutes at room temperature. Hydrochloride (4N) (3.73 ml) mixed with dioxane was added to the mixture, which was stirred for 1 hour and 30 minutes at the same temperature. The reaction solution was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (CH 2 Cl 2 : MeOH = 50: 1) to give N -phenyl- N- (3- (4- (piperazin-1--1-) as a final product. Carbonyl) phenyl) prop-2-ynyl) pentaneamide ( N- phenyl- N- (3- (4- (piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) was obtained (363 mg). , 60% yield).
1H NMR (CDCl3, 500MHz) δ 7.48-7.30(9H, m), 4.73(2H, s), 3.73-3.39(4H, br), 2.97-2.86(4H, br), 2.09-2.06(2H, t), 1.60-1.54(2H, m), 1.25-1.19(2H, m), 0.83-0.80(3H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.48-7.30 (9H, m), 4.73 (2H, s), 3.73-3.39 (4H, br), 2.97-2.86 (4H, br), 2.09-2.06 (2H, t), 1.60-1.54 (2H, m), 1.25-1.19 (2H, m), 0.83-0.80 (3H, t).
실시예Example 3.  3. NN -(3-(4-(4--(3- (4- (4- 메틸피페라진Methylpiperazine -1--One- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜탄아마이드Phenylpentaneamide ( ( NN -(3-(4-(4--(3- (4- (4- methylpiperazinemethylpiperazine -1-carbonyl)phenyl)prop-2--1-carbonyl) phenyl) prop-2- ynylynyl )-)- NN -phenylpentanamide) (LMT-692)의 제조-phenylpentanamide) (LMT-692)
상기 실시예 2에서 수득한 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (33.3 mg, 0.072 mmol)와 수산화칼륨 (KOH)(9.09 mg, 0.108 mmol)을 N,N-다이메틸포름아마이드 (DMF)(1 ml)에 녹인 후, 상온에서 5분 동안 교반하였다. 상기 혼합물에 아이오도메탄 (iodomethane)(9 μl, 0.144 mmol)을 가한 후, 같은 온도에서 17시간 동안 교반하였다. 반응액을 감압 하에 농축한 뒤, 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산 마그네슘 (MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피 (CH2Cl2:MeOH=20:1)로 정제하여 최종 생성물인 N-(3-(4-(4-메틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드(N-(3-(4-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다 (6 mg, 20% 수율). N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (33.3 mg, 0.072 mmol) obtained in Example 2 and potassium hydroxide ( KOH) (9.09 mg, 0.108 mmol) was dissolved in N , N -dimethylformamide (DMF) (1 ml) and stirred at room temperature for 5 minutes. Iodomethane (9 μl, 0.144 mmol) was added to the mixture, followed by stirring at the same temperature for 17 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (CH 2 Cl 2 : MeOH = 20: 1) to give N- (3- (4- (4-methylpiperazin-1-carbonyl) phenyl) prop- as the final product. 2-ynyl) - N - phenyl pentane amide (N - (3- (4- ( 4-methylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) - N -phenylpentanamide a) to give (6 mg, 20% yield).
1H NMR (CDCl3, 400MHz) δ 7.40-7.21(9H, m), 4.65(2H, s), 3.71-3.34(4H, br), 2.41-2.25(4H, br), 2.25(3H, s), 2.02-1.99(2H, t), 1.54-1.46(2H, m), 1.18-1.12(2H, m), 0.76-0.71(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.40-7.21 (9H, m), 4.65 (2H, s), 3.71-3.34 (4H, br), 2.41-2.25 (4H, br), 2.25 (3H, s) , 2.02-1.99 (2H, t), 1.54-1.46 (2H, m), 1.18-1.12 (2H, m), 0.76-0.71 (3H, t).
실시예Example 4.  4. NN -(3-(4-(4--(3- (4- (4- 에틸피페라진Ethyl piperazine -1--One- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜탄아마이드Phenylpentaneamide ( ( NN -(3-(4-(4--(3- (4- (4- EthylpiperazineEthylpiperazine -1-carbonyl)phenyl)prop-2--1-carbonyl) phenyl) prop-2- ynylynyl )-)- NN -phenylpentanamide) (LMT-695)의 제조-phenylpentanamide) (LMT-695)
상기 실시예 3에서 수득한 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (24.8 mg, 0.061 mmol)와 수산화칼륨(8.62 mg, 0.154 mmol)을 N,N-다이메틸포름아마이드 (DMF)(1 ml)에 녹인 후, 상온에서 5분 동안 교반하였다. 상기 혼합물에 아이오도에탄 (iodoethane)(20 μl, 0.246 mmol)을 가하고, 같은 온도에서 17시간 동안 교반하였다. 반응액을 감압 하에 농축한 뒤, 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘 (MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피 (CH2Cl2:MeOH = 20:1)로 정제하여 최종 생성물인 N-(3-(4-(4-에틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드(N-(3-(4-(4-Ethylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다 (17.9 mg, 68% 수율). N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (24.8 mg, 0.061 mmol) obtained in Example 3 and potassium hydroxide ( 8.62 mg, 0.154 mmol) was dissolved in N , N -dimethylformamide (DMF) (1 ml) and stirred at room temperature for 5 minutes. Iodoethane (20 μl, 0.246 mmol) was added to the mixture and stirred for 17 hours at the same temperature. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (CH 2 Cl 2 : MeOH = 20: 1) to give N- (3- (4- (4-ethylpiperazin-1-carbonyl) phenyl) prop- as the final product. 2-ynyl) - N - phenyl pentane amide (N - (3- (4- ( 4-Ethylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) - N -phenylpentanamide a) was obtained (17.9 mg, 68% yield).
1H NMR (CDCl3, 400MHz) δ 7.40-7.20(9H, m), 4.65(2H,s), 3.73-3.35(4H,br), 2.44-2.31(6H,m), 2.03-1.99(2H,t), 1.54-1.46(2H,m), 1.20-1.13(2H,m), 1.05-1.01(3H,t), 0.78-0.73(3H,t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.40-7.20 (9H, m), 4.65 (2H, s), 3.73-3.35 (4H, br), 2.44-2.31 (6H, m), 2.03-1.99 (2H, t), 1.54-1.46 (2H, m), 1.20-1.13 (2H, m), 1.05-1.01 (3H, t), 0.78-0.73 (3H, t).
실시예Example 5.  5. NN -(3-(4-(4--(3- (4- (4- 아이소프로필피페라진Isopropyl piperazine -1--One- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -페닐펜탄아마이드 (-Phenylpentaneamide ( NN -(3-(4-(4--(3- (4- (4- isopropylpiperazineisopropylpiperazine -1-carbonyl)phenyl)prop-2-ynyl)--1-carbonyl) phenyl) prop-2-ynyl)- NN -phenylpentanamide) (LMT-696)의 제조-phenylpentanamide) (LMT-696)
상기 실시예 2에서 수득한 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (106 mg, 0.263 mmol)와 탄산수소나트륨 (27 mg, 0.316 mmol)을 얼음에서 냉각시킨 후, N,N-다이메틸포름아마이드 (DMF)(2 ml)를 넣고 1시간 동안 교반하였다. 상기 혼합물에 2-아이오도프로판 (2-iodopropane)(30 μl, 0.316 mmol)을 가하고, 얼음을 제거한 뒤, 60oC에서 가열, 환류하여 24시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 감압 하에 농축한 뒤, 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA:MeOH:TEA=12:12:1:0.1)로 정제하여 최종 생성물인 N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드(N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다 (66.8 mg, 57% 수율). N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (106 mg, 0.263 mmol) and sodium hydrogencarbonate obtained in Example 2 After cooling (27 mg, 0.316 mmol) on ice, N , N -dimethylformamide (DMF) (2 ml) was added thereto and stirred for 1 hour. 2-iodopropane (30 μl, 0.316 mmol) was added to the mixture, ice was removed, and the mixture was heated and refluxed at 60 ° C. for 24 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA: MeOH: TEA = 12: 12: 1: 0.1) to give N- (3- (4- (4-isopropylpiperazin-1-carbonyl) as a final product. ) phenyl) prop-2-ynyl) - N - phenyl pentane amide (N - (3- (4- ( 4-isopropylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) - N -phenylpentanamide) to give the (66.8 mg, 57% yield).
1H NMR (CDCl3, 500MHz) δ 7.47-7.30(9H, m), 4.73(2H, s), 3.78-3.40(4H, br), 2.75-2.72(1H, m), 2.59-2.44(4H, br), 2.09-2.06(2H, t), 1.59-1.56(2H, m), 1.25-1.20(2H, m), 1.06-1.04(6H, d), 0.83-0.80(3H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.47-7.30 (9H, m), 4.73 (2H, s), 3.78-3.40 (4H, br), 2.75-2.72 (1H, m), 2.59-2.44 (4H, br), 2.09-2.06 (2H, t), 1.59-1.56 (2H, m), 1.25-1.20 (2H, m), 1.06-1.04 (6H, d), 0.83-0.80 (3H, t).
실시예Example 6.  6. NN -(3-(4-(4-(2-하이드록시에틸)피페라진-1--(3- (4- (4- (2-hydroxyethyl) piperazin-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2-이닐)--2-ynyl)- NN -페닐펜탄아마이드 (-Phenylpentaneamide ( NN -(3-(4-(4-(2-hydroxyethyl)-(3- (4- (4- (2-hydroxyethyl) piperazinepiperazine -1-carbonyl)phenyl)prop-2-ynyl)--1-carbonyl) phenyl) prop-2-ynyl)- NN -phenylpentanamide) (LMT-827)의 제조-phenylpentanamide) (LMT-827)
상기 실시예 2에서 수득한 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (56 mg, 0.139 mmol)와 탄산칼륨 (77 mg, 0.556 mmol)을 아세토나이트릴 (3 ml)에 녹인 후, 상온에서 5분 동안 교반하였다. 상기 혼합물에 2-브로모에탄올 (2-bromoethanol)(99 μl, 1.39 mmol)을 가하고, 60oC에서 가열, 환류하여 17시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 여과하여 고체를 제거한 후, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(CH2Cl2:MeOH=50:1)로 정제하여 최종 생성물인 N-(3-(4-(4-(2-하이드록시에틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드(N-(3-(4-(4-(2-hydroxyethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다 (52 mg, 84% 수율). N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (56 mg, 0.139 mmol) obtained in Example 2 and potassium carbonate ( 77 mg, 0.556 mmol) was dissolved in acetonitrile (3 ml) and stirred at room temperature for 5 minutes. 2-bromoethanol (99 μl, 1.39 mmol) was added to the mixture, and the mixture was heated and refluxed at 60 ° C. for 17 hours. The reaction solution was cooled to room temperature, filtered to remove solids, and then concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (CH 2 Cl 2 : MeOH = 50: 1) to give N- (3- (4- (4- (2-hydroxyethyl) piperazin-1-carbonyl as a final product. ) phenyl) prop-2-ynyl) - N - phenyl pentane amide - prop-2-ynyl (N (3- (4- (4- (2-hydroxyethyl) piperazine-1-carbonyl) phenyl)) - N - phenylpentanamide) was obtained (52 mg, 84% yield).
1H NMR (CDCl3, 500MHz) δ 7.48-7.30(9H, m), 4.73(2H, s), 3.79(2H, br), 3.66-3.64(2H, t), 3.43(2H, br), 2.60-2.46(7H, br), 2.10-2.07(2H, t), 1.59-1.56(2H, m), 1.25-1.22(2H, m), 0.83-0.80(3H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.48-7.30 (9H, m), 4.73 (2H, s), 3.79 (2H, br), 3.66-3.64 (2H, t), 3.43 (2H, br), 2.60 -2.46 (7H, br), 2.10-2.07 (2H, t), 1.59-1.56 (2H, m), 1.25-1.22 (2H, m), 0.83-0.80 (3H, t).
실시예Example 7.  7. NN -(3-(4-(4-(-(3- (4- (4- ( 사이클로프로필메틸Cyclopropylmethyl )피페라진-1-Piperazine-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2-이닐)--2-ynyl)- NN -페닐펜탄아마이드 (-Phenylpentaneamide ( NN -(3-(4-(4-(-(3- (4- (4- ( cyclopropylmethylcyclopropylmethyl )) piperazinepiperazine -1-carbonyl)phenyl)prop-2-ynyl)--1-carbonyl) phenyl) prop-2-ynyl)- NN -phenylpentanamide) (-phenylpentanamide) ( LMTLMT -- 828)의828 제조 Produce
상기 실시예 2에서 수득한 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (50 mg, 0.124 mmol)와 탄산칼륨(51 mg, 0.372 mmol)을 N,N-다이메틸포름아마이드 (DMF)(2 ml)에 녹인 후, 상온에서 5분 동안 교반하였다. 상기 혼합물에 사이클로프로필메틸 브로마이드 (15 μl, 0.145 mmol)를 가하고, 80oC에서 가열, 환류하여 4시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 여과하여 고체를 제거한 후, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(CH2Cl2:MeOH=50:1)로 정제하여 최종 생성물인 N-(3-(4-(4-(사이클로프로필메틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드(N-(3-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다(16 mg, 28% 수율). N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) obtained in Example 2 and potassium carbonate ( 51 mg, 0.372 mmol) was dissolved in N , N -dimethylformamide (DMF) (2 ml) and then stirred at room temperature for 5 minutes. Cyclopropylmethyl bromide (15 μl, 0.145 mmol) was added to the mixture, heated at 80 ° C., refluxed and stirred for 4 hours. The reaction solution was cooled to room temperature, filtered to remove solids, and then concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (CH 2 Cl 2 : MeOH = 50: 1) to give N- (3- (4- (4- (cyclopropylmethyl) piperazin-1-carbonyl) phenyl as a final product. ) prop-2-ynyl) - N - phenyl pentane amide (N - (3- (4- ( 4- (cyclopropylmethyl) piperazine-1-carbonyl) phenyl) prop-2-ynyl) - N -phenylpentanamide) to give (16 mg, 28% yield).
1H NMR (CDCl3,500MHz) δ 7.48-7.28(9H, m), 4.73(2H, s), 3.82-3.45(4H, br), 2.63-2.49(4H, br), 2.32-2.31(2H, d), 2.09-2.06(2H, t), 1.60-1.56(2H, m), 1.25-1.20(3H, m), 0.83-0.80(3H, t), 0.55-0.53(2H, m), 0.12-0.11(2H, m). 1 H NMR (CDCl 3 , 500 MHz) δ 7.48-7.28 (9H, m), 4.73 (2H, s), 3.82-3.45 (4H, br), 2.63-2.49 (4H, br), 2.32-2.31 (2H, d), 2.09-2.06 (2H, t), 1.60-1.56 (2H, m), 1.25-1.20 (3H, m), 0.83-0.80 (3H, t), 0.55-0.53 (2H, m), 0.12- 0.11 (2H, m).
실시예Example 8.  8. NN -(3-(4-(4--(3- (4- (4- 사이클로헥실피페라진Cyclohexylpiperazine -1--One- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -페닐펜탄아마이드 (-Phenylpentaneamide ( NN -(3-(4-(4--(3- (4- (4- cyclohexylpiperazinecyclohexylpiperazine -1-carbonyl)phenyl)prop-2-ynyl)--1-carbonyl) phenyl) prop-2-ynyl)- NN -phenylpentanamide) (LMT-830)의 제조-phenylpentanamide) (LMT-830)
상기 실시예 2에서 수득한 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (50 mg, 0.124 mmol)와 아이오도사이클로헥산 (19 μl, 0.145 mmol)을 이용하여 상기 실시예 7과 동일한 방법으로 최종 생성물인 N-(3-(4-(4-사이클로헥실피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드 (N-(3-(4-(4-cyclohexylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다 (20 mg, 33% 수율). N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) obtained in Example 2 and iodocyclo The final product, N- (3- (4- (4-cyclohexylpiperazin-1-carbonyl) phenyl) prop-2, in the same manner as in Example 7 using hexane (19 μl, 0.145 mmol) - ynyl) - N-phenyl-pentane amide (N - (3- (4- ( 4-cyclohexylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) - N -phenylpentanamide) was obtained (20 mg, 33% yield ).
1H NMR (CDCl3, 500MHz) δ 7.48-7.31(9H, m), 4.73(2H, s), 3.77-3.39(4H, br), 2.63-2.49(4H, br), 2.31-2.28(1H, m), 2.09-2.06(2H, m), 1.91-1.79(4H, m), 1.65-1.54(3H, m), 1.28-1.16(6H, m), 1.13-1.08(1H, m), 0.83-0.80(3H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.48-7.31 (9H, m), 4.73 (2H, s), 3.77-3.39 (4H, br), 2.63-2.49 (4H, br), 2.31-2.28 (1H, m), 2.09-2.06 (2H, m), 1.91-1.79 (4H, m), 1.65-1.54 (3H, m), 1.28-1.16 (6H, m), 1.13-1.08 (1H, m), 0.83- 0.80 (3H, t).
실시예Example 9.  9. NN -(3-(4-(4-(-(3- (4- (4- ( 사이클로헥실메틸Cyclohexylmethyl )피페라진-1-Piperazine-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2-이닐)--2-ynyl)- NN -페닐펜탄아마이드 (-Phenylpentaneamide ( NN -(3-(4-(4-(-(3- (4- (4- ( cyclohexylmethylcyclohexylmethyl )) piperazinepiperazine -1-carbonyl)phenyl)prop-2-ynyl)--1-carbonyl) phenyl) prop-2-ynyl)- NN -phenylpentanamide) (LMT-831)의 제조-phenylpentanamide) (LMT-831)
상기 실시예 2에서 수득한 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (50 mg, 0.124 mmol)와 브로모메틸사이클로헥산 (20 μl, 0.145 mmol)을 이용하여 상기 실시예 7과 동일한 방법으로 최종 생성물인 N-(3-(4-(4-(사이클로헥실메틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드 (N-(3-(4-(4-(cyclohexylmethyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다 (35 mg, 56% 수율). N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) and bromomethyl obtained in Example 2 above Using cyclohexane (20 μl, 0.145 mmol) in the same manner as in Example 7, the final product, N- (3- (4- (4- (cyclohexylmethyl) piperazin-1-carbonyl) phenyl) prop profile-2-ynyl) - N - phenyl pentane amide (N - (3- (4- ( 4- (cyclohexylmethyl) piperazine-1-carbonyl) phenyl) prop-2-ynyl) - N -phenylpentanamide a) to give ( 35 mg, 56% yield).
1H NMR (CDCl3, 500MHz) δ 7.48-7.28(9H, m), 4.73(2H, s), 3.76-3.38(4H, br), 2.45-2.31(4H, br), 2.15-2.13(2H, m), 2.09-2.06(2H, m), 1.77-1.66(5H, m), 1.59-1.56(2H, m), 1.47-1.45(1H, m), 1.25-1.17(5H, m), 0.90-0.80(5H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.48-7.28 (9H, m), 4.73 (2H, s), 3.76-3.38 (4H, br), 2.45-2.31 (4H, br), 2.15-2.13 (2H, m), 2.09-2.06 (2H, m), 1.77-1.66 (5H, m), 1.59-1.56 (2H, m), 1.47-1.45 (1H, m), 1.25-1.17 (5H, m), 0.90- 0.80 (5H, t).
실시예Example 10.  10. NN -(3-(4-(4--(3- (4- (4- 아이소부틸피페라진Isobutyl Piperazine -1--One- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -페닐펜탄아마이드 (-Phenylpentaneamide ( NN -(3-(4-(4--(3- (4- (4- isobutylpiperazineisobutylpiperazine -1-carbonyl)phenyl)prop-2-ynyl)--1-carbonyl) phenyl) prop-2-ynyl)- NN -phenylpentanamide) (LMT-832)의 제조-phenylpentanamide) (LMT-832)
상기 실시예 2에서 수득한 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (50 mg, 0.124 mmol)와 1-아이오도-2-메틸프로판 (17 μl, 0.145 mmol)을 이용하여 상기 실시예 7과 동일한 방법으로 최종 생성물인 N-(3-(4-(4-아이소부틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드 (N-(3-(4-(4-isobutylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다(34 mg, 60% 수율). N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) obtained in Example 2 and 1-Io The final product, N- (3- (4- (4-isobutylpiperazin-1-carbonyl) phenyl), in the same manner as in Example 7 using Fig. 2-methylpropane (17 μl, 0.145 mmol) prop-2-ynyl) - N - phenyl pentane amide (N - (3- (4- ( 4-isobutylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) - N -phenylpentanamide a) was obtained (34 mg , 60% yield).
1H NMR (CDCl3, 500MHz)δ 7.47-7.28(9H, m), 4.73(2H, s), 3.76-3.39(4H, br), 2.46-2.32(4H, br), 2.11-2.07(4H, m), 1.79-1.76(1H, m), 1.59-1.56(2H, m), 1.25-1.20(2H, m), 0.91-0.89(6H, d), 0.83-0.80(3H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.47-7.28 (9H, m), 4.73 (2H, s), 3.76-3.39 (4H, br), 2.46-2.32 (4H, br), 2.11-2.07 (4H, m), 1.79-1.76 (1H, m), 1.59-1.56 (2H, m), 1.25-1.20 (2H, m), 0.91-0.89 (6H, d), 0.83-0.80 (3H, t).
실시예Example 11.  11. NN -페닐--Phenyl- NN -(3-(4-(4-(-(3- (4- (4- ( 프로프Prof -2--2- 이닐Inil )피페라진-1-Piperazine-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2-이닐)펜탄아마이드 (-2-ynyl) pentaneamide ( NN -phenyl--phenyl- NN -(3-(4-(4-(prop-2--(3- (4- (4- (prop-2- ynylynyl )) piperazinepiperazine -1-carbonyl)phenyl)prop-2-ynyl)pentanamide) (LMT-833)의 제조Preparation of -1-carbonyl) phenyl) prop-2-ynyl) pentanamide) (LMT-833)
상기 실시예 2에서 수득한 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (50 mg, 0.124 mmol)와 탄산칼륨 (51 mg, 0.372 mmol)을 N,N-다이메틸포름아마이드 (DMF)(2 ml)에 녹인 후, 상온에서 5분 동안 교반하였다. 상기 혼합물에 프로파길 브로마이드 (12 μl, 0.145 mmol)를 가하고, 상온에서 17시간 동안 교반하였다. 반응액을 여과하여 고체를 제거한 후, 감압 하에 농축한 뒤, 상기 농축액을 실리카겔 컬럼 크로마토그래피(CH2Cl2:MeOH=100:1)로 정제하여 최종 생성물인 N-페닐-N-(3-(4-(4-(프로프-2-이닐)피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (N-phenyl-N-(3-(4-(4-(prop-2-ynyl)piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide)를 수득하였다(28 mg, 51% 수율). N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) obtained in Example 2 and potassium carbonate ( 51 mg, 0.372 mmol) was dissolved in N , N -dimethylformamide (DMF) (2 ml) and then stirred at room temperature for 5 minutes. Propargyl bromide (12 μl, 0.145 mmol) was added to the mixture and stirred at room temperature for 17 hours. The reaction solution was filtered to remove solids, concentrated under reduced pressure, and the concentrate was purified by silica gel column chromatography (CH 2 Cl 2 : MeOH = 100: 1) to give N -phenyl- N- (3- as the final product. (4- (4- (prop-2-ynyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide ( N- phenyl- N- (3- (4- (4- ( prop-2-ynyl) piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) was obtained (28 mg, 51% yield).
1H NMR (CDCl3, 500MHz) δ 7.48-7.28(9H, m), 4.73(2H, s), 3.82-3.44(4H, br), 3.36(2H, s), 2.65-2.51(4H, br), 2.30(1H, s), 2.10-2.07(2H, m), 1.60-1.54(2H, m), 1.26-1.19(2H, m), 0.83-0.80(3H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.48-7.28 (9H, m), 4.73 (2H, s), 3.82-3.44 (4H, br), 3.36 (2H, s), 2.65-2.51 (4H, br) , 2.30 (1H, s), 2.10-2.07 (2H, m), 1.60-1.54 (2H, m), 1.26-1.19 (2H, m), 0.83-0.80 (3H, t).
실시예Example 12.  12. NN -(3-(4-(4--(3- (4- (4- 시아노피페라진Cyanopiperazine -1--One- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜탄아마이드Phenylpentaneamide ( ( NN -(3-(4-(4--(3- (4- (4- cyanopiperazinecyanopiperazine -1-carbonyl)phenyl)prop-2--1-carbonyl) phenyl) prop-2- ynylynyl )-)- NN -phenylpentanamide) (LMT-829)의 제조 -phenylpentanamide) (LMT-829)
상기 실시예 2에서 수득한 N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (50 mg, 0.124 mmol)와 트리메틸실릴 시아나이드 (49 μl, 0.372 mmol)를 아세토나이트릴 (2 ml)에 녹인 후, 상온에서 5분 동안 교반하였다. 상기 혼합물에 나트륨 하이포클로라이트 (43 μl, 0.620 mmol)를 가하고, 80℃ 에서 가열, 환류하여 12시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 여과하여 고체를 제거한 후, 감압 하에 농축하였다. 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘 (MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(CH2Cl2:MeOH=200:1)로 정제하여 최종 생성물인 N-(3-(4-(4-시아노피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드 (N-(3-(4-(4-cyanopiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다(11 mg, 21% 수율). N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) and trimethylsilyl cya obtained in Example 2 above Nide (49 μl, 0.372 mmol) was dissolved in acetonitrile (2 ml) and stirred at room temperature for 5 minutes. Sodium hypochlorite (43 μl, 0.620 mmol) was added to the mixture, and the mixture was heated and refluxed at 80 ° C. for 12 hours. The reaction solution was cooled to room temperature, filtered to remove solids, and then concentrated under reduced pressure. The obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (CH 2 Cl 2 : MeOH = 200: 1) to give N- (3- (4- (4-cyanopiperazin-1-carbonyl) phenyl) prop as the final product. 2-ynyl) -N -phenylpentanamide ( N- (3- (4- (4-cyanopiperazine-1-carbonyl) phenyl) prop-2-ynyl) -N- phenylpentanamide) was obtained (11 mg, 21 % Yield).
1H NMR (CDCl3,500MHz) δ 7.48-7.30(9H, m), 4.73(2H, s), 3.81-3.26(8H, br), 2.09-2.06(2H, t), 1.60-1.54(2H, m), 1.25-1.19(2H, m), 0.83-0.80(3H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.48-7.30 (9H, m), 4.73 (2H, s), 3.81-3.26 (8H, br), 2.09-2.06 (2H, t), 1.60-1.54 (2H, m), 1.25-1.19 (2H, m), 0.83-0.80 (3H, t).
실시예Example 13.  13. terttert -- 부틸 4-(4-(3-(Butyl 4- (4- (3- ( NN -(3--(3- 플루오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )벤조일)피페라진-1-카복실레이트 () Benzoyl) piperazine-1-carboxylate ( terttert -butyl 4-(4-(3-(-butyl 4- (4- (3- ( NN -(3-fluorophenyl)pentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate) (-(3-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) ( LMTLMT -884)의 제조-884)
단계 1 : Step 1: NN -(3--(3- 플루오로페닐Fluorophenyl )) 펜탄아마이드Pentaneamide ( ( NN -(3--(3- fluorophenylfluorophenyl )) pentanamidepentanamide )의 제조Manufacturing
3-플루오로아닐린(3-fluoroaniline)(0.87 ml, 9.00 mmol)과 발레로일 클로라이드(valeroyl chloride)(2.18 ml, 18.00 mmol)를 이용하여 N-(3-플루오로페닐)펜탄아마이드 (N-(3-fluorophenyl)pentanamide)를 수득하였다(1.74 g, 99% 수율).3-fluoro-aniline (3-fluoroaniline) (0.87 ml , 9.00 mmol) and using a chloride (valeroyl chloride) (2.18 ml, 18.00 mmol) in ballet N - (3-fluorophenyl) pentane amide (N - (3-fluorophenyl) pentanamide) was obtained (1.74 g, 99% yield).
단계 2 : Step 2: terttert -- 부틸 4-(4-(3-(Butyl 4- (4- (3- ( NN -(3--(3- 플루오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )벤조일)피페라진-1-카복실레이트 () Benzoyl) piperazine-1-carboxylate ( terttert -butyl 4-(4-(3-(-butyl 4- (4- (3- ( NN -(3-fluorophenyl)pentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate)의 제조Preparation of-(3-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
상기 단계 1에서 수득한 N-(3-플루오로페닐)펜탄아마이드 (101 mg, 0.519 mmol)와 상기 실시예 1의 단계 4에서 수득한 tert-부틸 4-(4-(3-(메틸설포닐옥시)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (329 mg, 0.779 mmol)를 이용하여 상기 실시예 1의 단계 5와 동일한 방법으로 최종 생성물인 tert -부틸 4-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (tert-butyl 4-(4-(3-(N-(3-fluorophenyl)pentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate)를 수득하였다 (197 mg, 73% 수율). N- (3-fluorophenyl) pentaneamide (101 mg, 0.519 mmol) obtained in step 1 and tert -butyl 4- (4- (3- (methylsulfonyl) obtained in step 4 of Example 1 Oxy) prop-1-ynyl) benzoyl) piperazine-1-carboxylate (329 mg, 0.779 mmol) in the same manner as in step 5 of Example 1 above, the final product tert - butyl 4- (4- (3- ( N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate ( tert -butyl 4- (4- (3- ( N- (3 -fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (197 mg, 73% yield).
1H NMR (CDCl3, 400MHz) δ 7.44-7.32(5H, m), 7.12-7.06(3H, m), 4.72(2H, s), 3.73-3.38(8H, br), 2.10-2.07(2H, t), 1.59-1.57(2H, m), 1.47(9H, s), 1.23-1.20(2H, m), 0.83-0.80(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.44-7.32 (5H, m), 7.12-7.06 (3H, m), 4.72 (2H, s), 3.73-3.38 (8H, br), 2.10-2.07 (2H, t), 1.59-1.57 (2H, m), 1.47 (9H, s), 1.23-1.20 (2H, m), 0.83-0.80 (3H, t).
실시예Example 14.  14. NN -(3--(3- 플루오로페닐Fluorophenyl )-)- NN -(3-(4-(피페라진-1--(3- (4- (piperazin-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2-이닐)펜탄아마이드 (-2-ynyl) pentaneamide ( NN -(3--(3- fluorophenylfluorophenyl )-)- NN -(3-(4-(-(3- (4- ( piperazinepiperazine -1-carbonyl)phenyl)prop-2-ynyl)pentanamide) (LMT-885)의 제조Preparation of -1-carbonyl) phenyl) prop-2-ynyl) pentanamide) (LMT-885)
상기 실시예 13에서 수득한 tert -부틸 4-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (200 mg, 0.383 mmol)를 이용하여 상기 실시예 12와 동일한 방법으로 최종 생성물인 N-(3-플루오로페닐)-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (N-(3-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide)를 수득하였다 (94 mg, 58% 수율). Tert - butyl 4- (4- (3- ( N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate (200) obtained in Example 13 mg, 0.383 mmol), example 12, and the final product in the same manner using N - (3-fluorophenyl) - N - (3- (4- (piperazine-1-carbonyl) phenyl) prop- 2-ynyl) pentane amide (N - (3-fluorophenyl) - N - (3- (4- (piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide a) was obtained (94 mg, 58% yield).
1H NMR (CDCl3, 400MHz) δ 7.45-7.32(5H, m), 7.15-7.07(3H, m), 4.72(2H, s), 3.75-3.37(4H, br), 2.94-2.80(4H, br), 2.10-2.07(2H, t), 1.89(1H, br), 1.60-1.57(2H, m), 1.25-1.24(2H, m), 0.84-0.82(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.45-7.32 (5H, m), 7.15-7.07 (3H, m), 4.72 (2H, s), 3.75-3.37 (4H, br), 2.94-2.80 (4H, br), 2.10-2.07 (2H, t), 1.89 (1H, br), 1.60-1.57 (2H, m), 1.25-1.24 (2H, m), 0.84-0.82 (3H, t).
실시예Example 15.  15. NN -(3--(3- 플루오로페닐Fluorophenyl )-)- NN -(3-(4-(4--(3- (4- (4- 아이소프로필피페라진Isopropyl piperazine -1--One- 카보닐Carbonyl )페닐)프로프-2-이닐)펜탄아마이드 () Phenyl) prop-2-ynyl) pentaneamide ( NN -(3--(3- fluorophenylfluorophenyl )-)- NN -(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide) (-(3- (4- (4-isopropylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) ( LMTLMT -- 886)의886 제조 Produce
상기 실시예 14에서 수득한 N-(3-플루오로페닐)-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (46 mg, 0.109 mmol)와 2-아이오도프로판 (2-iodopropane)(0.375 ml, 3.77 mmol)을 이용하여 최종 생성물인 N-(3-플루오로페닐)-N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드(N-(3-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide)를 수득하였다 (20 mg, 40% 수율).A N obtained in Example 14 - (3-fluorophenyl) - N - (3- (4- (piperazine-1-carbonyl) phenyl) prop-2-ynyl), pentane amide (46 mg, 0.109 mmol) and 2-iodopropane (0.375 ml, 3.77 mmol) to give the final product N- (3-fluorophenyl) -N- (3- (4- (4-isopropylpipepe) piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide (N - (3-fluorophenyl) - N - (3- (4- (4-isopropylpiperazine-1-carbonyl) prop-2- phenyl) ynyl) pentanamide) was obtained (20 mg, 40% yield).
1H NMR (CDCl3, 400MHz) δ 7.44-7.32(5H, m), 7.15-7.06(3H, m), 4.72(2H, s), 3.78-3.41(4H, br), 2.75-2.72(1H, m), 2.59-2.45(4H, br), 2.11-2.09(2H, t), 1.60-1.57(2H, m), 1.25-1.22(2H, m), 1.06-1.05(6H, d), 0.85-0.82(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.44-7.32 (5H, m), 7.15-7.06 (3H, m), 4.72 (2H, s), 3.78-3.41 (4H, br), 2.75-2.72 (1H, m), 2.59-2.45 (4H, br), 2.11-2.09 (2H, t), 1.60-1.57 (2H, m), 1.25-1.22 (2H, m), 1.06-1.05 (6H, d), 0.85- 0.82 (3H, t).
실시예Example 16.  16. terttert -부틸 4-(4-(3-(-Butyl 4- (4- (3- ( NN -(4--(4- 플루오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )벤조일)피페라진-1-카복실레이트((Benzoyl) piperazine-1-carboxylate ( terttert -butyl 4-(4-(3-(-butyl 4- (4- (3- ( NN -(4-fluorophenyl)pentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate) (-(4-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) ( LMTLMT -839)의 제조-839)
단계 1 : Step 1: NN -- (4-플루오로페닐)펜탄아마이드((4-fluorophenyl) pentaneamide ( NN -(4-fluorophenyl)pentanamide-(4-fluorophenyl) pentanamide )의 제조Manufacturing
4-플루오로아닐린 (4-fluoroaniline)(85 μl, 0.90 mmol)과 발레로일 클로라이드(valeroyl chloride)(0.22 ml, 1.80 mmol)를 이용하여 상기 실시예 1의 단계 1과 동일한 방법으로 N-(4-플루오로페닐)펜탄아마이드(N-(4-fluorophenyl)pentanamide)를 수득하였다 (174 mg, 99% 수율).With an aniline (4-fluoroaniline) (85 μl , 0.90 mmol) and ballet chloride (valeroyl chloride) (0.22 ml, 1.80 mmol) with 4-fluoro in the same manner as in Step 1 of Example 1 N - ( 4-fluorophenyl) pentanamide ( N- (4-fluorophenyl) pentanamide) was obtained (174 mg, 99% yield).
단계 2 : Step 2: terttert -부틸 4-(4-(3-(-Butyl 4- (4- (3- ( NN -(4--(4- 플루오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )벤조일)피페라진-1-카복실레이트 () Benzoyl) piperazine-1-carboxylate ( terttert -butyl 4-(4-(3-(-butyl 4- (4- (3- ( NN -(4-fluorophenyl)pentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate)의 제조Preparation of-(4-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
상기 단계 1에서 수득한 N-(4-플루오로페닐)펜탄아마이드(1.17 g, 6.00 mmol)와 상기 실시예 1의 단계 4에서 수득한 tert-부틸 4-(4-(3-(메틸설포닐옥시)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (3.80 g, 8.99 mmol)를 이용하여 상기 실시예 1의 단계 5와 동일한 방법으로 최종 생성물인 tert-부틸 4-(4-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트(tert-butyl 4-(4-(3-(N-(4-fluorophenyl)pentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate)를 수득하였다 (2.28 g, 73% 수율). N- (4-fluorophenyl) pentaneamide (1.17 g, 6.00 mmol) obtained in step 1 above and tert -butyl 4- (4- (3- (methylsulfonyl) obtained in step 4 of Example 1 Oxy) prop-1-ynyl) benzoyl) piperazine-1-carboxylate (3.80 g, 8.99 mmol) in the same manner as in step 5 of Example 1 above The final product tert -butyl 4- (4- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate ( tert -butyl 4- (4- (3- ( N- (4-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (2.28 g, 73% yield).
1H NMR (CDCl3, 400MHz) δ 7.39-7.29(6H, m), 7.17-7.14(2H, m), 4.71(2H, s), 3.73-3.38(8H, br), 2.07-2.04(2H, t), 1.60-1.54(2H, m), 1.47(9H, s), 1.25-1.19(2H, m), 0.84-0.81(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.39-7.29 (6H, m), 7.17-7.14 (2H, m), 4.71 (2H, s), 3.73-3.38 (8H, br), 2.07-2.04 (2H, t), 1.60-1.54 (2H, m), 1.47 (9H, s), 1.25-1.19 (2H, m), 0.84-0.81 (3H, t).
실시예Example 17.  17. N-N- (4-(4- 플루오로페닐Fluorophenyl )-)- NN -(3-(4-(피페라진-1--(3- (4- (piperazin-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2-이닐)펜탄아마이드 (-2-ynyl) pentaneamide ( N-N- (4-(4- fluorophenylfluorophenyl )-)- NN -(3-(4-(-(3- (4- ( piperazinepiperazine -1-carbonyl)phenyl)prop-2-ynyl)pentanamide) (-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) ( LMTLMT -- 840)의840 제조 Produce
상기 실시예 16에서 수득한 tert-부틸 4-(4-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (2.28 g, 4.37 mmol)를 이용하여 상기 실시예 12와 동일한 방법으로 최종 생성물인 N-(4-플루오로페닐)-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드(N-(4-fluorophenyl)-N-(3-(4-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide)를 수득하였다 (1.06 g, 58% 수율). Tert -butyl 4- (4- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate (2.28) g, 4.37 mmol) of the above-described N final product in the same manner as in example 12 using (4-fluorophenyl) - N - (3- (4- (piperazine-1-carbonyl) phenyl) prop- 2-ynyl) pentane amide (N - (4-fluorophenyl) - N - (3- (4- (piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) was obtained (1.06 g, 58% yield).
1H NMR (CDCl3, 400MHz) δ 7.38-7.30(6H, m), 7.17-7.15(2H, m), 4.71(2H, s), 3.77-3.40(4H, br), 2.96-2.79(5H, br), 2.06-2.03(2H, t), 1.57-1.54(2H, m), 1.25-1.22(2H, m), 0.84-0.82(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.38-7.30 (6H, m), 7.17-7.15 (2H, m), 4.71 (2H, s), 3.77-3.40 (4H, br), 2.96-2.79 (5H, br), 2.06-2.03 (2H, t), 1.57-1.54 (2H, m), 1.25-1.22 (2H, m), 0.84-0.82 (3H, t).
실시예Example 18.  18. NN -(4--(4- 플루오로페닐Fluorophenyl )-)- NN -(3-(4-(4--(3- (4- (4- 아이소프로필피페라진Isopropyl piperazine -1--One- 카보닐Carbonyl )페닐)프로프-2-이닐)펜탄아마이드 () Phenyl) prop-2-ynyl) pentaneamide ( NN -(4--(4- fluorophenylfluorophenyl )-)- NN -(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide) (-(3- (4- (4-isopropylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) ( LMTLMT -- 841)의841) 제조 Produce
상기 실시예 17에서 수득한 N-(4-플루오로페닐)-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (1.06 g, 2.51 mmol)와 2-아이오도프로판 (2-iodopropane)(0.375 ml, 3.77 mmol)을 이용하여 상기 실시예 15와 동일한 방법으로 최종 생성물인 N-(4-플루오로페닐)-N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드(N-(4-fluorophenyl)-N-(3-(4-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide)를 수득하였다 (465 mg, 40% 수율).The embodiment one N obtained in Example 17 - (4-fluorophenyl) - N - (3- (4- (piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide (1.06 g, 2.51 mmol) and 2-iodopropane (0.375 ml, 3.77 mmol) in the same manner as in Example 15 above, where the final product was N- (4-fluorophenyl) -N- (3- ( 4- (4-isopropyl-piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentane amide (N - (4-fluorophenyl) - N - (3- (4- (4-isopropylpiperazine-1- carbonyl) phenyl) prop-2-ynyl) pentanamide) was obtained (465 mg, 40% yield).
1H NMR (CDCl3, 400MHz) δ 7.37-7.28(6H, m), 7.17-7.13(2H, m), 4.71(2H, s), 3.78-3.40(4H, br), 2.75-2.72(1H, m), 2.59-2.45(4H, br), 2.07-2.04(2H, t), 1.58-1.55(2H, m), 1.25-1.21(2H, m), 1.06-1.04(6H, d), 0.84-0.81(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.37-7.28 (6H, m), 7.17-7.13 (2H, m), 4.71 (2H, s), 3.78-3.40 (4H, br), 2.75-2.72 (1H, m), 2.59-2.45 (4H, br), 2.07-2.04 (2H, t), 1.58-1.55 (2H, m), 1.25-1.21 (2H, m), 1.06-1.04 (6H, d), 0.84- 0.81 (3H, t).
실시예Example 19.  19. NN -(3-(4-(-(3- (4- ( 몰폴린Morpholine -4--4- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜탄아마이드Phenylpentaneamide ( ( NN -(3-(4-(-(3- (4- ( morpholinemorpholine -4-carbonyl)phenyl)prop-2--4-carbonyl) phenyl) prop-2- ynylynyl )-)- NN -- phenylpentanamidephenylpentanamide ) (LMT-682)의 제조) (LMT-682) Preparation
단계 1 : Step 1: (4-브로모페닐)(몰폴리노)메타논(4-bromophenyl) (morpholino) methanone ((4-bromophenyl)(morpholino)methanone)((4-bromophenyl) (morpholino) methanone)
4-브로모벤조산(4-bromobenzoic acid) (340 mg, 1.70 mmol)과 몰폴린 (0.18 ml, 2.04 mmol)을 이용하여 상기 실시예 1의 단계 2와 동일한 방법으로 (4-브로모페닐)(몰폴리노)메타논 ((4-bromophenyl)(morpholino)methanone)을 수득하였다 (455 mg, 수율 99%).(4-bromophenyl) (4-bromobenzoic acid) (340 mg, 1.70 mmol) and morpholine (0.18 ml, 2.04 mmol) in the same manner as in step 2 of Example 1 above Morpholino) methanone ((4-bromophenyl) (morpholino) methanone) was obtained (455 mg, 99% yield).
단계 2 : Step 2: (4-(3-하이드록시프로프-1-이닐)페닐)(몰폴리노)메타논(4- (3-hydroxyprop-1-ynyl) phenyl) (morpholino) methanone ((4-(3-hydroxyprop-1-ynyl)phenyl)(morpholino)methanone)의 제조Preparation of ((4- (3-hydroxyprop-1-ynyl) phenyl) (morpholino) methanone)
상기 단계 1에서 수득한 (4-브로모페닐)(몰폴리노)메타논 (455 mg, 1.68 mmol)과 프로파길 알코올 (propagyl alcohol)(0.196 ml, 3.36 mmol)을 이용하여 상기 실시예 1의 단계 3과 동일한 방법으로 (4-(3-하이드록시프로프-1-이닐)페닐)(몰폴리노)메타논((4-(3-hydroxyprop-1-ynyl)phenyl)(morpholino)methanone)을 수득하였다(371 mg, 수율 90%).Example 4 was prepared using (4-bromophenyl) (morpholino) methanone (455 mg, 1.68 mmol) and propagyl alcohol (0.196 ml, 3.36 mmol) obtained in step 1. (4- (3-hydroxyprop-1-ynyl) phenyl) (morpholino) methanone ((4- (3-hydroxyprop-1-ynyl) phenyl) (morpholino) methanone) Was obtained (371 mg, yield 90%).
단계 step 3 : 33: 3 -(4-(-(4-( 몰폴린Morpholine -4--4- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil 메탄설포네이트Methanesulfonate (3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl methanesulfonate)의 제조Preparation of (3- (4- (morpholine-4-carbonyl) phenyl) prop-2-ynyl methanesulfonate)
상기 단계 2에서 수득한 (4-(3-하이드록시프로프-1-이닐)페닐)(몰폴리노)메타논 (371 mg, 1.51 mmol)과 메탄설포닐 클로라이드 (0.128 ml, 1.66 mmol)를 이용하여 상기 실시예 1의 단계 4와 동일한 방법으로 3-(4-(몰폴린-4-카보닐)페닐)프로프-2-이닐 메탄설포네이트(3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl methanesulfonate)를 수득하였다 (391 mg, 수율 80%).(4- (3-hydroxyprop-1-ynyl) phenyl) (morpholino) methanone (371 mg, 1.51 mmol) and methanesulfonyl chloride (0.128 ml, 1.66 mmol) obtained in step 2 were prepared. 3- (4- (morpholine-4-carbonyl) phenyl) prop-2-ynyl methanesulfonate (3- (4- (morpholine-4-carbonyl) in the same manner as in Step 4 of Example 1 above ) phenyl) prop-2-ynyl methanesulfonate) was obtained (391 mg, yield 80%).
단계 4 : Step 4: NN -(3-(4-(-(3- (4- ( 몰폴린Morpholine -4--4- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜탄아마이Phenylpentane amy 드(De ( NN -(3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl)--(3- (4- (morpholine-4-carbonyl) phenyl) prop-2-ynyl)- NN -phenylpentanamide)의 제조-phenylpentanamide)
상기 실시예 1의 단계 1에서 수득한 N-페닐펜탄아마이드(143 mg, 0.81 mmol)와 상기 단계 3에서 수득한 3-(4-(몰폴린-4-카보닐)페닐)프로프-2-이닐 메탄설포네이트(391 mg, 1.21 mmol)를 이용하여 상기 실시예 1의 단계 5와 동일한 방법으로 최종 생성물인 N-(3-(4-(몰폴린-4-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드 (N-(3-(4-(morpholine-4-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다 (371 mg, 수율 90%). N -phenylpentaneamide (143 mg, 0.81 mmol) obtained in step 1 of Example 1 and 3- (4- (morpholine-4-carbonyl) phenyl) prop-2- obtained in step 3 above. The final product N- (3- (4- (morpholin-4-carbonyl) phenyl) prop- in the same manner as in Step 5 of Example 1 using inyl methanesulfonate (391 mg, 1.21 mmol) 2-ynyl) -N -phenylpentanamide ( N- (3- (4- (morpholine-4-carbonyl) phenyl) prop-2-ynyl) -N- phenylpentanamide) was obtained (371 mg, yield 90%). .
1H NMR (400 MHz, CDCl3) δ 7.48-7.28(m, 9H), 4.73(s, 2H), 3.74-3.66(br, 6 H), 3.43(br, 2H), 2.08(m, 2H), 1.57(m, 2H), 1.22 (m, 2H), 0.81(t, 3H). 1 H NMR (400 MHz, CDCl 3 ) δ 7.48-7.28 (m, 9H), 4.73 (s, 2H), 3.74-3.66 (br, 6H), 3.43 (br, 2H), 2.08 (m, 2H) , 1.57 (m, 2H), 1.22 (m, 2H), 0.81 (t, 3H).
실시예Example 20.  20. NN -페닐--Phenyl- NN -(3-(4-(피페리딘-1--(3- (4- (piperidine-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )) 펜탄아마이드Pentaneamide ( ( NN -phenyl--phenyl- NN -(3-(4-(-(3- (4- ( piperidinepiperidine -1-carbonyl)phenyl)prop-2-ynyl)pentanamide) (LMT-683)의 제조Preparation of -1-carbonyl) phenyl) prop-2-ynyl) pentanamide) (LMT-683)
단계 1 : Step 1: (4-브로모페닐)(피페리딘-1-일)메타논(4-bromophenyl) (piperidin-1-yl) methanone ((4-((4- bromophenylbromophenyl )() ( piperidinpiperidin -1-yl)methanone))의 제조-1-yl) methanone))
4-브로모벤조산 (4-bromobenzoic acid)(318 mg, 1.58 mmol)과 피페리딘 (piperidine)(0.21 ml, 1.90 mmol)을 이용하여 상기 실시예 1의 단계 2와 동일한 방법으로 (4-브로모페닐)(피페리딘-1-일)메타논 ((4-bromophenyl)(piperidin-1-yl)methanone))을 수득하였다 (458.2 mg, 100% 수율).Using 4-bromobenzoic acid (318 mg, 1.58 mmol) and piperidine (0.21 ml, 1.90 mmol) in the same manner as in step 2 of Example 1 (4-bromobenzoic acid) (4-bromobenzoic acid) Mophenyl) (piperidin-1-yl) methanone ((4-bromophenyl) (piperidin-1-yl) methanone)) was obtained (458.2 mg, 100% yield).
단계 2 : (4-(3-Step 2: (4- (3- 하이드록시프로프Hydroxyprop -1--One- 이닐Inil )페닐)피리딘-1-일)) Phenyl) pyridin-1-yl) 메타논Metanon ((4-(3-hydroxyprop-1-ynyl)phenyl)(piperidin-1-yl)methanone)의 제조Preparation of ((4- (3-hydroxyprop-1-ynyl) phenyl) (piperidin-1-yl) methanone)
상기 단계 1에서 수득한 (4-브로모페닐)(피페리딘-1-일)메타논 (458.2 mg, 1.71 mmol)과 프로파길 알코올 (propagyl alcohol)(0.199 ml, 3.42 mmol)을 이용하여 상기 실시예 1의 단계 3과 동일한 방법으로 (4-(3-하이드록시프로프-1-이닐)페닐)피리딘-1-일)메타논 ((4-(3-hydroxyprop-1-ynyl)phenyl)(piperidin-1-yl)methanone)을 수득하였다(374 mg, 수율 90%).Using (4-bromophenyl) (piperidin-1-yl) methanone (458.2 mg, 1.71 mmol) and propagyl alcohol (0.199 ml, 3.42 mmol) obtained in step 1, (4- (3-hydroxyprop-1-ynyl) phenyl) pyridin-1-yl) methanone ((4- (3-hydroxyprop-1-ynyl) phenyl) in the same manner as in step 3 of Example 1 (piperidin-1-yl) methanone) was obtained (374 mg, yield 90%).
단계 step 3 : 33: 3 -(4-(피리딘-1--(4- (pyridine-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil 메탄설포네이트Methanesulfonate (3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl methanesulfonate)의 제조Preparation of (3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl methanesulfonate)
상기 단계 2에서 수득한 (4-(3-하이드록시프로프-1-이닐)페닐)피리딘-1-일)메타논 (374 mg, 1.54 mmol)과 메탄설포닐 클로라이드 (0.131 ml, 1.69 mmol)를 이용하여 상기 실시예 1의 단계 4와 동일한 방법으로 3-(4-(피리딘-1-카보닐)페닐)프로프-2-이닐 메탄설포네이트 (3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl methanesulfonate)를 수득하였다 (396 mg, 수율 80%).(4- (3-hydroxyprop-1-ynyl) phenyl) pyridin-1-yl) methanone (374 mg, 1.54 mmol) and methanesulfonyl chloride (0.131 ml, 1.69 mmol) obtained in step 2 above. 3- (4- (pyridine-1-carbonyl) phenyl) prop-2-ynyl methanesulfonate (3- (4- (piperidine-1-carbonyl) in the same manner as in Step 4 of Example 1 above using ) phenyl) prop-2-ynyl methanesulfonate) was obtained (396 mg, yield 80%).
단계 4 : Step 4: NN -페닐--Phenyl- NN -(3-(4-(피페리딘-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드(-(3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide ( NN -phenyl--phenyl- NN -(3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide)의 제조Preparation of-(3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide)
상기 실시예 1의 단계 1에서 수득한 N-페닐펜탄아마이드 (146 mg, 0.82 mmol)와 상기 단계 3에서 수득한 3-(4-(피리딘-1-카보닐)페닐)프로프-2-이닐 메탄설포네이트 (396 mg, 1.23 mmol)를 이용하여 상기 실시예 1의 단계 5와 동일한 방법으로 최종 생성물인 N-페닐-N-(3-(4-(피페리딘-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (N-phenyl-N-(3-(4-(piperidine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide)를 수득하였다 (50 mg, 15% 수율). N -phenylpentanamide (146 mg, 0.82 mmol) obtained in step 1 of Example 1 and 3- (4- (pyridine-1-carbonyl) phenyl) prop-2-ynyl obtained in step 3 above. The final product, N -phenyl- N- (3- (4- (piperidine-1-carbonyl) phenyl, in the same manner as in Step 5 of Example 1 above using methanesulfonate (396 mg, 1.23 mmol) ) Prop-2-ynyl) pentaneamide ( N- phenyl- N- (3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) was obtained (50 mg, 15% yield). ).
1H NMR (CDCl3, 400MHz) δ 7.40-7.13(9H, m), 4.65(2H, s, CH2), 3.62-3.24(4H, br), 2.02-1.99(2H,t), 1.60(4H, br), 1.52-1.46(2H, m), 1.44 (2H, br), 1.20-1.08 (2H, m), 0.78-0.73(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.40-7.13 (9H, m), 4.65 (2H, s, CH 2 ), 3.62-3.24 (4H, br), 2.02-1.99 (2H, t), 1.60 (4H , br), 1.52-1.46 (2H, m), 1.44 (2H, br), 1.20-1.08 (2H, m), 0.78-0.73 (3H, t).
실시예Example 21.  21. NN ,, NN -- 다이에틸Diethyl -4-(3-(-4- (3- ( NN -- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1--One- 이닐Inil )) 벤즈아마이Benz Amai 드 (De ( NN ,, NN -diethyl-4-(3-(-diethyl-4- (3- ( NN -- phenylpentanamidophenylpentanamido )prop-1-prop-1- ynylynyl )) benzamidebenzamide ) () ( LMTLMT -- 883)의883) 제조 Produce
단계 step 1 : 41: 4 -- 브로모Bromo -- NN ,, NN -- 다이에틸벤즈아마이드Diethylbenzamide (4-(4- bromobromo -- NN ,, NN -diethylbenzamide)의 제조-diethylbenzamide)
4-브로모벤조산(4-bromobenzoic acid)(800 mg, 3.98 mmol)과 다이에틸아민(diethylamine)(0.49 ml, 4.78 mmol)을 이용하여 상기 실시예 1의 단계 2와 동일한 방법으로 4-브로모-N,N-다이에틸벤즈아마이드 (4-bromo-N,N-diethylbenzamide)를 수득하였다 (700 mg, 69% 수율).4-bromobenzoic acid (4-bromobenzoic acid) (800 mg, 3.98 mmol) and diethylamine (0.49 ml, 4.78 mmol) using 4-bromo in the same manner as in step 2 of Example 1 above N , N -diethylbenzamide (4-bromo- N , N- diethylbenzamide) was obtained (700 mg, 69% yield).
단계 2 : Step 2: NN ,, NN -- 다이에틸Diethyl -4--4- (3-하이드록시프로프-1-이닐)벤즈아마이드(3-hydroxyprop-1-ynyl) benzamide (( NN ,, NN -diethyl-4-(3-hydroxyprop-1-ynyl)benzamide)의 제조Preparation of -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide)
상기 단계 1에서 수득한 4-브로모-N,N-다이에틸벤즈아마이드 (700 mg, 2.73 mmol)와 프로파길 알코올 (propagyl alcohol)(0.32 ml, 5.47 mmol)을 이용하여 상기 실시예 1의 단계 3과 동일한 방법으로 N,N-다이에틸-4-(3-하이드록시프로프-1-이닐)벤즈아마이드(N,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzamide)를 수득하였다 (425 mg, 67% 수율). Step 1 of Example 1 using 4-bromo- N , N -diethylbenzamide (700 mg, 2.73 mmol) and propagyl alcohol (0.32 ml, 5.47 mmol) obtained in step 1 above. In the same manner as 3, N , N -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide was obtained. ( N , N- diethyl-4- (3-hydroxyprop-1-ynyl) benzamide) (425 mg, 67% yield).
단계 step 3 : 33: 3 -(4-(-(4-( 다이에틸카바모일Diethylcarbamoyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil 메탄설포네이트Methanesulfonate (3-(4-(diethylcarbamoyl)phenyl)prop-2-ynyl methanesulfonate)의 제조Preparation of (3- (4- (diethylcarbamoyl) phenyl) prop-2-ynyl methanesulfonate)
상기 단계 2에서 수득한 N,N-다이에틸-4-(3-하이드록시프로프-1-이닐)벤즈아마이드 (425 mg, 1.84 mmol)와 메탄설포닐 클로라이드 (0.16 ml, 2.02 mmol)를 이용하여 상기 실시예 1의 단계 4와 동일한 방법으로 3-(4-(다이에틸카바모일)페닐)프로프-2-이닐 메탄설포네이트 (3-(4-(diethylcarbamoyl)phenyl)prop-2-ynyl methanesulfonate)를 수득하였다(483 mg, 85% 수율).Using N , N -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide (425 mg, 1.84 mmol) and methanesulfonyl chloride (0.16 ml, 2.02 mmol) obtained in step 2 above. 3- (4- (diethylcarbamoyl) phenyl) prop-2-ynyl methanesulfonate (3- (4- (diethylcarbamoyl) phenyl) prop-2-ynyl in the same manner as in Step 4 of Example 1 above methanesulfonate) was obtained (483 mg, 85% yield).
단계 4 : Step 4: NN ,, NN -- 다이에틸Diethyl -4--4- (3-((3- ( NN -페닐펜탄아미도)프로프-1-이닐)벤즈아마이-Phenylpentaneamido) prop-1-ynyl) benzamy 드(De ( NN ,, NN -diethyl-4-(3-(-diethyl-4- (3- ( NN -phenylpentanamido)prop-1-ynyl)benzamide)의 제조-phenylpentanamido) prop-1-ynyl) benzamide)
상기 실시예 1의 단계 1에서 수득한 N-페닐펜탄아마이드 (94 mg, 0.530 mmol)와 상기 단계 3에서 수득한 수득한 3-(4-(다이에틸카바모일)페닐)프로프-2-이닐 메탄설포네이트(246 mg, 0.795 mmol)를 이용하여 상기 실시예 1의 단계 5와 동일한 방법으로 최종 생성물인 N,N-다이에틸-4-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤즈아마이드 (N,N-diethyl-4-(3-(N-phenylpentanamido)prop-1-ynyl)benzamide)를 수득하였다 (81 mg, 39% 수율). N -phenylpentanamide (94 mg, 0.530 mmol) obtained in step 1 of Example 1 and 3- (4- (diethylcarbamoyl) phenyl) prop-2-ynyl obtained in step 3 above. Using methanesulfonate (246 mg, 0.795 mmol) in the same manner as in Step 5 of Example 1, the final product, N , N -diethyl-4- (3- ( N -phenylpentaneamido) prop- 1-ynyl) benzamide ( N , N- diethyl-4- (3- ( N- phenylpentanamido) prop-1-ynyl) benzamide) was obtained (81 mg, 39% yield).
1H NMR (CDCl3, 400MHz) δ 7.48-7.30(9H, m), 4.73(2H, s), 3.53-3.23(4H, br), 2.10-2.07(2H, t), 1.59-1.57(2H, m), 1.23-1.10(8H, m), 0.83-0.80(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.48-7.30 (9H, m), 4.73 (2H, s), 3.53-3.23 (4H, br), 2.10-2.07 (2H, t), 1.59-1.57 (2H, m), 1.23-1.10 (8H, m), 0.83-0.80 (3H, t).
실시예Example 22.  22. NN -페닐--Phenyl- NN -(3-(3-(피페라진-1--(3- (3- (piperazin-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )) 펜탄아마이드Pentaneamide ( ( NN -phenyl--phenyl- NN -(3-(3-(-(3- (3- ( piperazinepiperazine -1-carbonyl)phenyl)prop-2-ynyl)pentanamide) (LMT-837)의 제조 Preparation of -1-carbonyl) phenyl) prop-2-ynyl) pentanamide) (LMT-837)
단계 1 : Step 1: terttert -부틸 4-(3--Butyl 4- (3- 브로모벤조일Bromobenzoyl )피페라진-1-Piperazine-1- 카복실레이트(Carboxylate ( terttert -butyl -butyl 4-(3-bromobenzoyl)piperazine-1-carboxylate)의 제조Preparation of 4- (3-bromobenzoyl) piperazine-1-carboxylate)
3-브로모벤조산 (3-bromobenzoic acid)(1.50 g, 7.46 mmol)과 tert-부틸 피페라진-1-카복실레이트 (tert-butyl piperazine-1-carboxylate)(1.67 g, 8.95 mmol)를 이용하여 상기 실시예 1의 단계 2와 동일한 방법으로 tert-부틸 4-(3-브로모벤조일)피페라진-1-카복실레이트 (tert-butyl 4-(3-bromobenzoyl)piperazine-1-carboxylate)를 수득하였다 (2.75 g, 99% 수율).3-bromo-benzoate (3-bromobenzoic acid) (1.50 g, 7.46 mmol) and tert - using butyl piperazine-1-carboxylate (tert -butyl piperazine-1-carboxylate ) (1.67 g, 8.95 mmol) the In the same manner as in step 2 of Example 1, tert -butyl 4- (3-bromobenzoyl) piperazine-1-carboxylate was obtained ( tert- butyl 4- (3-bromobenzoyl) piperazine-1-carboxylate) ( 2.75 g, 99% yield).
단계 2 : Step 2: terttert -부틸 4-(3-(3--Butyl 4- (3- (3- 하이드록시프로프Hydroxyprop -1--One- 이닐Inil )) 벤조일Benzoyl )피페라진-1-Piperazine-1- 카복실레이트Carboxylate (( terttert -butyl 4-(3-(3--butyl 4- (3- (3- hydroxyprophydroxyprop -1--One- ynylynyl )) benzoylbenzoyl )) piperazinepiperazine -1-carboxylate)의 제조-1-carboxylate)
상기 단계 1에서 수득한 tert-부틸 4-(3-브로모벤조일)피페라진-1-카복실레이트 (2.75 g, 7.45 mmol)와 프로파길 알코올 (propagyl alcohol)(0.87 ml, 14.89 mmol)을 이용하여 상기 실시예 1의 단계 3과 동일한 방법으로 tert-부틸 4-(3-(3-하이드록시프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (tert-butyl 4-(3-(3-hydroxyprop-1-ynyl)benzoyl)piperazine-1-carboxylate)를 수득하였다 (2.32 g, 90% 수율). Tert -butyl 4- (3-bromobenzoyl) piperazine-1-carboxylate (2.75 g, 7.45 mmol) and propagyl alcohol (0.87 ml, 14.89 mmol) obtained in step 1 were used. in the same manner as in step 3 in example 1 tert - butyl 4- (3- (3-hydroxy-prop-1-ynyl) benzoyl) piperazine-l-carboxylate (tert -butyl 4- (3- ( 3-hydroxyprop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (2.32 g, 90% yield).
단계 3 : Step 3: terttert -부틸 4-(3-(3-(-Butyl 4- (3- (3- ( 메틸설포닐옥시Methylsulfonyloxy )) 프로프Prof -1--One- 이닐Inil )) 벤조일Benzoyl )피페라진-1-카복실레이트 (Piperazine-1-carboxylate ( terttert -butyl 4-(3-(3-(-butyl 4- (3- (3- ( methylsulfonyloxymethylsulfonyloxy )prop-1-ynyl)benzoyl)piperazine-1-carboxylate)의 제조) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
상기 단계 2에서 수득한 tert-부틸 4-(3-(3-하이드록시프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (2.32 g, 6.75 mmol)와 메탄설포닐 클로라이드 (0.58 ml, 7.42 mmol)를 이용하여 상기 실시예 1의 단계 4와 동일한 방법으로 tert-부틸 4-(3-(3-(메틸설포닐옥시)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (tert-butyl 4-(3-(3-(methylsulfonyloxy)prop-1-ynyl)benzoyl)piperazine-1-carboxylate)를 수득하였다 (1.78 g, 63% 수율). Tert -butyl 4- (3- (3-hydroxyprop-1-ynyl) benzoyl) piperazine-1-carboxylate (2.32 g, 6.75 mmol) and methanesulfonyl chloride (0.58 ml) obtained in step 2 above. , 7.42 mmol) in the same manner as in Step 4 of Example 1, tert -butyl 4- (3- (3- (methylsulfonyloxy) prop-1-ynyl) benzoyl) piperazine-1-carboxyl Yield ( tert- butyl 4- (3- (3- (methylsulfonyloxy) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (1.78 g, 63% yield).
단계 4 : Step 4: terttert -부틸 4-(3-(3-(-Butyl 4- (3- (3- ( NN -- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1--One- 이닐Inil )) 벤조일Benzoyl )피페라진-1-카복실레이트(Piperazine-1-carboxylate ( terttert -butyl 4-(3-(3-(-butyl 4- (3- (3- ( NN -- phenylpentanamidophenylpentanamido )prop-1-ynyl)benzoyl)piperazine-1-carboxylate)의 제조) prop-1-ynyl) benzoyl) piperazine-1-carboxylate)
상기 실시예 1의 단계 1에서 수득한 N-페닐펜탄아마이드 (275 mg, 1.55 mmol)와 상기 단계 3에서 수득한 tert-부틸 4-(3-(3-(메틸설포닐옥시)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (982 mg, 2.33 mmol)를 이용하여 상기 실시예 1의 단계 5와 동일한 방법으로 최종 생성물인 tert-부틸 4-(3-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트(tert-butyl 4-(3-(3-(N-phenylpentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate)를 수득하였다 (323 mg, 70% 수율). N -phenylpentaneamide (275 mg, 1.55 mmol) obtained in step 1 of Example 1 and tert -butyl 4- (3- (3- (methylsulfonyloxy) prop-1 obtained in step 3) -Inyl) benzoyl) piperazine-1-carboxylate (982 mg, 2.33 mmol) in the same manner as in Step 5 of Example 1, tert -butyl 4- (3- (3- ( N- Phenylpentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate ( tert -butyl 4- (3- (3- ( N -phenylpentanamido) prop-1-ynyl) benzoyl) piperazine-1- carboxylate) (323 mg, 70% yield).
단계 5: Step 5: NN -페닐--Phenyl- NN -(3-(3-(피페라진-1--(3- (3- (piperazin-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )) 펜탄아마이드Pentaneamide ( ( NN -phenyl--phenyl- NN -(3-(3-(-(3- (3- ( piperazinepiperazine -1-carbonyl)phenyl)prop-2--1-carbonyl) phenyl) prop-2- ynylynyl )) pentanamidepentanamide ) (LMT-837)의 제조 ) (LMT-837) Preparation
상기 단계 4에서 수득한 tert-부틸 4-(3-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (323 mg, 0.62 mmol)를 이용하여 상기 실시예 2과 동일한 방법으로 최종 생성물인 N-페닐-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드(N-phenyl-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide)를 수득하였다 (152mg, 58% 수율). Tert -butyl 4- (3- (3- ( N -phenylpentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate (323 mg, 0.62 mmol) obtained in step 4 was used. of the final product in the same manner as example 2 N - phenyl - N - (3- (3- (piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide (N -phenyl- N -(3- (3- (piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) was obtained (152 mg, 58% yield).
1H NMR (CDCl3, 400MHz) δ 7.45-7.26(9H, m), 4.69(2H, s), 3.75-3.36(4H, br), 2.94-2.80(4H, br), 2.59(1H, br), 2.07-2.04(2H, t), 1.56-1.53(2H, m), 1.22-1.18(2H, m), 0.80-0.77(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.45-7.26 (9H, m), 4.69 (2H, s), 3.75-3.36 (4H, br), 2.94-2.80 (4H, br), 2.59 (1H, br) , 2.07-2.04 (2H, t), 1.56-1.53 (2H, m), 1.22-1.18 (2H, m), 0.80-0.77 (3H, t).
실시예Example 23.  23. NN -(3-(3-(4-메틸피페라진-1-카보닐)페닐)프로프-2-이닐)--(3- (3- (4-methylpiperazin-1-carbonyl) phenyl) prop-2-ynyl)- NN -페닐펜탄아마이드(-Phenylpentaneamide ( NN -(3-(3-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)--(3- (3- (4-methylpiperazine-1-carbonyl) phenyl) prop-2-ynyl)- NN -phenylpentanamide) (LMT-838)의 제조-phenylpentanamide) (LMT-838)
상기 실시예 22에서 수득한 N-페닐-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (50 mg, 0.124 mmol)와 포름알데하이드(37% in H2O)(1.5ml)를 포름산(2.0 ml)에 녹인 후, 100oC에서 가열, 환류하여 4시간 동안 교반하였다. 반응액을 감압 하에 농축한 뒤, 수산화나트륨 수용액(2.0 M)을 가하여 적정하였다. 이 후, 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산 마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(CH2Cl2:MeOH=20:1)로 정제하여 N-(3-(3-(4-메틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드(N-(3-(3-(4-methylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다(24 mg, 46% 수율). N -phenyl- N- (3- (3- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (50 mg, 0.124 mmol) obtained in Example 22 and formaldehyde ( 37% in H 2 O) (1.5 ml) was dissolved in formic acid (2.0 ml), heated at 100 ° C., refluxed and stirred for 4 hours. The reaction solution was concentrated under reduced pressure, and then titrated by addition of aqueous sodium hydroxide solution (2.0 M). Thereafter, the mixture was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (CH 2 Cl 2 : MeOH = 20: 1) to obtain N- (3- (3- (4- (methylpiperazin-1-carbonyl) phenyl) prop-2-ynyl ) -N -phenylpentanamide ( N- (3- (3- (4-methylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) -N- phenylpentanamide) was obtained (24 mg, 46% yield).
1H NMR (CDCl3, 400MHz) δ 7.46-7.29(9H, m), 4.70(2H, s), 3.79-3.39(4H, br), 2.48-2.32(7H, br), 2.08-2.05(2H, t), 1.57-1.54(2H, m), 1.23-1.19(2H, m), 0.81-0.78(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.46-7.29 (9H, m), 4.70 (2H, s), 3.79-3.39 (4H, br), 2.48-2.32 (7H, br), 2.08-2.05 (2H, t), 1.57-1.54 (2H, m), 1.23-1.19 (2H, m), 0.81-0.78 (3H, t).
실시예Example 24.  24. NN -(3-(3-(4--(3- (3- (4- 아이소프로필피페라진Isopropyl piperazine -1--One- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -페닐펜탄아마이드 (-Phenylpentaneamide ( NN -(3-(3-(4--(3- (3- (4- isopropylpiperazineisopropylpiperazine -1-carbonyl)phenyl)prop-2-ynyl)--1-carbonyl) phenyl) prop-2-ynyl)- NN -phenylpentanamide) (LMT-842)의 제조-phenylpentanamide) (LMT-842)
상기 실시예 22에서 수득한 N-페닐-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (152 mg, 0.36 mmol)와 2-아이오도프로판 (2-iodopropane)(90 μl, 0.90 mmol)을 이용하여 상기 실시예 5와 동일한 방법으로 최종 생성물인 N-(3-(3-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드 (N-(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다 (67 mg, 40% 수율). N -phenyl- N- (3- (3- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide (152 mg, 0.36 mmol) obtained in Example 22 and 2-Io The final product, N- (3- (3- (4-isopropylpiperazin-1-carbonyl) phenyl, was prepared in the same manner as in Example 5 using 2-iodopropane (90 μl, 0.90 mmol). ) prop-2-ynyl) - N - phenyl pentane amide (N - (3- (3- ( 4-isopropylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) - N -phenylpentanamide a) to yield (67 mg, 40% yield).
1H NMR (CDCl3, 400MHz) δ 7.47-7.29(9H, m), 4.71(2H, s), 3.79-3.40(4H, br), 2.78-2.75(1H, m), 2.60-2.46(4H, br), 2.09-2.06(2H, t), 1.58-1.55(2H, m), 1.24-1.20(2H, m), 1.07-1.05(6H, d), 0.82-0.79(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.47-7.29 (9H, m), 4.71 (2H, s), 3.79-3.40 (4H, br), 2.78-2.75 (1H, m), 2.60-2.46 (4H, br), 2.09-2.06 (2H, t), 1.58-1.55 (2H, m), 1.24-1.20 (2H, m), 1.07-1.05 (6H, d), 0.82-0.79 (3H, t).
실시예Example 25.  25. terttert -부틸-4-(3-(3-(-Butyl-4- (3- (3- ( NN -(4--(4- 플루오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )벤조일)피페라진-1-카복실레이트 () Benzoyl) piperazine-1-carboxylate ( terttert -butyl 4-(3-(3-(-butyl 4- (3- (3- ( NN -(4-fluorophenyl)pentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate) (-(4-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) ( LMTLMT -887)의 제조-887)
상기 실시예 16의 단계 1에서 수득한 N-(4-플루오로페닐)펜탄아마이드 (275 mg, 1.41 mmol)와 상기 실시예 22의 단계 3에서 수득한 tert-부틸 4-(3-(3-(메틸설포닐옥시)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (894 mg, 2.12 mmol)를 이용하여 상기 실시예 1의 단계 5와 동일한 방법으로 최종 생성물인 tert-부틸-4-(3-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (tert-butyl 4-(3-(3-(N-(4-fluorophenyl)pentanamido)prop-1-ynyl)benzoyl)piperazine-1-carboxylate)를 수득하였다 (625 mg, 85% 수율). N- (4-fluorophenyl) pentaneamide (275 mg, 1.41 mmol) obtained in Step 1 of Example 16 and tert -butyl 4- (3- (3- (Methylsulfonyloxy) prop-1-ynyl) benzoyl) piperazine-1-carboxylate (894 mg, 2.12 mmol) in the same manner as in Step 5 of Example 1 above, the final product tert -butyl- 4- (3- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazin-1-carboxylate ( tert -butyl 4- (3- (3- ( N- (4-fluorophenyl) pentanamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate) was obtained (625 mg, 85% yield).
1H NMR (CDCl3, 400MHz) δ 7.33-7.20(6H, m), 7.09-7.06(2H, m), 4.62(2H, s), 3.66-3.31(8H, br), 2.00-1.97(2H, t), 1.52-1.49(2H, m), 1.47(9H, s), 1.18-1.13(2H, m), 0.76-0.73(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.33-7.20 (6H, m), 7.09-7.06 (2H, m), 4.62 (2H, s), 3.66-3.31 (8H, br), 2.00-1.97 (2H, t), 1.52-1.49 (2H, m), 1.47 (9H, s), 1.18-1.13 (2H, m), 0.76-0.73 (3H, t).
실시예Example 26.  26. NN -(4--(4- 플루오로페닐Fluorophenyl )-)- NN -(3-(3-(피페라진-1--(3- (3- (piperazin-1- 카보닐Carbonyl )페닐)) Phenyl) 프로프Prof -2-이닐)펜탄아마이드 (-2-ynyl) pentaneamide ( NN -(4--(4- fluorophenylfluorophenyl )-)- NN -(3-(3-(-(3- (3- ( piperazinepiperazine -1-carbonyl)phenyl)prop-2-ynyl)pentanamide) (LMT-888)의 제조 Preparation of -1-carbonyl) phenyl) prop-2-ynyl) pentanamide) (LMT-888)
상기 실시예 25에서 수득한 tert-부틸-4-(3-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트 (762 mg, 1.46 mmol)를 이용하여 상기 실시예 2과 동일한 방법으로 최종 생성물인 N-(4-플루오로페닐)-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (N-(4-fluorophenyl)-N-(3-(3-(piperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide)를 수득하였다 (356mg, 58% 수율). Tert -butyl-4- (3- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate obtained in Example 25 ( 762 mg, 1.46 mmol) in example 2 with the final product the N in the same manner by using the - (4-fluorophenyl) - N - (3- (3- (piperazine-1-carbonyl) phenyl) pro P -2-ynyl) pentaneamide ( N- (4-fluorophenyl) -N- (3- (3- (piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) was obtained (356 mg, 58%). yield).
1H NMR (CDCl3, 400MHz) δ 7.39-7.29(6H, m), 7.17-7.14(2H, m), 4.70(2H, s), 3.77-3.39(4H, br), 2.98-2.85(4H, br), 2.08-2.05(2H, t), 1.60-1.54(2H, m), 1.25-1.19(2H, m), 0.84-0.81(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.39-7.29 (6H, m), 7.17-7.14 (2H, m), 4.70 (2H, s), 3.77-3.39 (4H, br), 2.98-2.85 (4H, br), 2.08-2.05 (2H, t), 1.60-1.54 (2H, m), 1.25-1.19 (2H, m), 0.84-0.81 (3H, t).
실시예Example 27.  27. NN -(4--(4- 플루오로페닐Fluorophenyl )-)- NN -(3-(3-(4--(3- (3- (4- 아이소프로필피페라진Isopropyl piperazine -1--One- 카보닐Carbonyl )페닐)프로프-2-이닐)펜탄아마이드 () Phenyl) prop-2-ynyl) pentaneamide ( NN -(4--(4- fluorophenylfluorophenyl )-)- NN -(3-(3-(4-isopropylpiperazine-1-carbonyl)phenyl)prop-2-ynyl)pentanamide) (-(3- (3- (4-isopropylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) pentanamide) ( LMTLMT -- 889)의889) 제조 Produce
상기 실시예 26에서 수득한 N-(4-플루오로페닐)-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드 (150 mg, 0.356 mmol)와 2-아이오도프로판 (2-iodopropane)(53 μl, 0.534 mmol)을 이용하여 상기 실시예 5와 동일한 방법으로 최종 생성물인 N-(4-플루오로페닐)-N-(3-(3-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드를 수득하였다 (66 mg, 40% 수율).A N obtained in Example 26 - (4-fluorophenyl) - N - (3- (3- (piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide (150 mg, 0.356 mmol) and 2-iodopropane (53 μl, 0.534 mmol) in the same manner as in Example 5, where the final product, N- (4-fluorophenyl) -N- (3- ( 3- (4-isopropylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide was obtained (66 mg, 40% yield).
1H NMR (CDCl3, 400MHz) δ 7.37-7.29(6H, m), 7.16-7.13(2H, m), 4.70(2H, s), 3.79-3.39(4H, br), 2.75-2.73(1H, m), 2.59-2.45(4H, br), 2.07-2.04(2H, t), 1.58-1.55(2H, m), 1.25-1.21(2H, m), 1.06-1.05(6H, d), 0.84-0.81(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.37-7.29 (6H, m), 7.16-7.13 (2H, m), 4.70 (2H, s), 3.79-3.39 (4H, br), 2.75-2.73 (1H, m), 2.59-2.45 (4H, br), 2.07-2.04 (2H, t), 1.58-1.55 (2H, m), 1.25-1.21 (2H, m), 1.06-1.05 (6H, d), 0.84- 0.81 (3H, t).
실시예Example 28.  28. NN -(3-(4--(3- (4- 하이드록시페닐Hydroxyphenyl )) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜탄아마이드Phenylpentaneamide ( ( NN -(3-(4-hydroxyphenyl)prop-2-ynyl)--(3- (4-hydroxyphenyl) prop-2-ynyl)- NN -phenylpentanamide) (LMT-890)의 제조-phenylpentanamide) (LMT-890)
단계 1 : Step 1: NN -(프로프-2-이닐)아닐린 (-(Prop-2-ynyl) aniline ( NN -(prop-2-ynyl)aniline)의 제조Preparation of-(prop-2-ynyl) aniline)
아닐린 (2.94 ml, 32.21 mmol)과 탄산칼륨 (4.90 g, 35.43 mmol)을 아세토나이트릴 (40 ml)에 녹인 후, 5분 동안 교반하였다. 상기 혼합물에 프로파길 브로마이드 (propargyl bromide)(3.05 ml, 35.43 mmol)를 가하고, 상온에서 17시간 동안 교반하였다. 반응액을 여과하여 고체를 제거한 후, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피 (Hex)로 정제하여 N-(프로프-2-이닐)아닐린(N-(prop-2-ynyl)aniline)를 수득하였다 (2.23 g, 53% 수율).Aniline (2.94 ml, 32.21 mmol) and potassium carbonate (4.90 g, 35.43 mmol) were dissolved in acetonitrile (40 ml) and stirred for 5 minutes. Propargyl bromide (3.05 ml, 35.43 mmol) was added to the mixture, followed by stirring at room temperature for 17 hours. The reaction solution was filtered to remove solids and then concentrated under reduced pressure. Purification of the concentrate by silica gel column chromatography (Hex) N - (prop-2-ynyl) aniline (N - (prop-2- ynyl) aniline) was obtained (2.23 g, 53% yield).
단계 2 : Step 2: NN -페닐--Phenyl- NN -- (프로프-2-이닐)펜탄아마이드(Prop-2-ynyl) pentaneamide (( NN -phenyl--phenyl- NN -(prop-2-ynyl)pentanamide)의 제조Preparation of-(prop-2-ynyl) pentanamide)
상기 단계 1에서 수득한 N-(프로프-2-이닐)아닐린 (828 mg, 6.31 mmol)과 발레로일 클로라이드 (valeroyl chloride)(1.53 ml, 12.62 mmol)를 이용하여 상기 실시예 1의 단계 1과 동일한 방법으로 N-페닐-N-(프로프-2-이닐)펜탄아마이드 (N-phenyl-N-(prop-2-ynyl)pentanamide)를 수득하였다 (1.29 g, 95% 수율). Step 1 of Example 1 using N- (prop-2-ynyl) aniline (828 mg, 6.31 mmol) and valeroyl chloride (1.53 ml, 12.62 mmol) obtained in step 1 above. in the same manner as N - phenyl - N - (prop-2-ynyl) pentane amide (N -phenyl- N - (prop- 2-ynyl) pentanamide) was obtained (1.29 g, 95% yield).
단계 3 : Step 3: NN -(3-(4--(3- (4- 하이드록시페닐Hydroxyphenyl )) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜탄아마이드Phenylpentaneamide (( NN -(3-(4-hydroxyphenyl)prop-2-ynyl)--(3- (4-hydroxyphenyl) prop-2-ynyl)- NN -phenylpentanamide)의 제조-phenylpentanamide)
상기 단계 2에서 수득한 N-페닐-N-(프로프-2-이닐)펜탄아마이드(550 mg, 2.55 mmol)와 4-아이오도페놀 (4-iodophenol)(422 mg, 1.92 mmol)을 트리에틸아민(15 ml)에 녹인 후, 5분 동안 교반하였다. 상기 혼합물에 비스(트리페닐포스핀)팔라듐(II) 다이클로라이드(89 mg, 0128 mmol)와 요오드화구리(I) (49 mg, 0.255 mmol)를 가하고 50oC에서 가열, 환류하여 5시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 감압 하에 농축한 뒤, 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산 마그네슘 (MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=10:1)로 정제하여 최종 생성물인 N-(3-(4-하이드록시페닐)프로프-2-이닐)-N-페닐펜탄아마이드(N-(3-(4-hydroxyphenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다 (590 mg, 75% 수율).Triethyl N -phenyl- N- (prop-2-ynyl) pentaneamide (550 mg, 2.55 mmol) and 4-iodophenol (422 mg, 1.92 mmol) obtained in step 2 were obtained. Dissolved in amine (15 ml) and stirred for 5 minutes. Bis (triphenylphosphine) palladium (II) dichloride (89 mg, 0128 mmol) and copper iodide (I) (49 mg, 0.255 mmol) were added to the mixture, heated at 50 ° C., refluxed and stirred for 5 hours. It was. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 10: 1 ) to give the final product of N - (3- (4-hydroxyphenyl) prop-2-ynyl) - N-phenyl-pentane amide (N - (3- (4-hydroxyphenyl) prop-2-ynyl) -N- phenylpentanamide) was obtained (590 mg, 75% yield).
1H NMR (CDCl3, 400MHz) δ 8.15(1H, br), 7.47-6.83(9H, m), 4.66(2H, s), 2.13-2.10(2H, t), 1.60-1.54(2H, m), 1.22-1.17(2H, m), 0.80-0.77(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 8.15 (1H, br), 7.47-6.83 (9H, m), 4.66 (2H, s), 2.13-2.10 (2H, t), 1.60-1.54 (2H, m) , 1.22-1.17 (2H, m), 0.80-0.77 (3H, t).
실시예Example 29. 2-(4-(3-( 29. 2- (4- (3- ( NN -- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1--One- 이닐Inil )) 페녹시Phenoxy )아세트산 (2-(4-(3-(Acetic acid (2- (4- (3- ( NN -phenylpentanamido)prop-1-ynyl)phenoxy)acetic acid) (LMT-891)의 제조-phenylpentanamido) prop-1-ynyl) phenoxy) acetic acid) (LMT-891)
단계 1 : 에틸 2-(4-(3-(Step 1: ethyl 2- (4- (3- ( NN -- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1--One- 이닐Inil )) 페녹시Phenoxy )아세테이트 (ethyl (2-(4-(3-(Acetate (ethyl (2- (4- (3- ( NN -phenylpentanamido)prop-1-ynyl)phenoxy)acetate)의 제조-phenylpentanamido) prop-1-ynyl) phenoxy) acetate)
상기 실시예 28에서 수득한 N-(3-(4-하이드록시페닐)프로프-2-이닐)-N-페닐펜탄아마이드 (590 mg, 1.92 mmol)와 탄산칼륨 (796 mg, 5.76 mmol)을 아세토나이트릴(15 ml)에 녹인 후, 30분 동안 교반하였다. 상기 혼합물에 에틸 브로모아세테이트 (ethyl bromoacetate)(0.23 ml, 2.11 mmol)를 가하고, 상온에서 17시간 동안 교반하였다. 반응액을 여과하여 고체를 제거한 후, 감압 하에 농축한 뒤, 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=10:1)로 정제하여 에틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)페녹시)아세테이트 (ethyl 2-(4-(3-(N-phenylpentanamido)prop-1-ynyl)phenoxy)acetate)를 수득하였다 (530 mg, 70% 수율). N- (3- (4-hydroxyphenyl) prop-2-ynyl) -N -phenylpentanamide (590 mg, 1.92 mmol) and potassium carbonate (796 mg, 5.76 mmol) obtained in Example 28 were prepared. It was dissolved in acetonitrile (15 ml) and stirred for 30 minutes. Ethyl bromoacetate (0.23 ml, 2.11 mmol) was added to the mixture, followed by stirring at room temperature for 17 hours. The reaction solution was filtered to remove solids, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 10: 1) to obtain ethyl 2- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) phenoxy) acetate (ethyl 2 -(4- (3- ( N- phenylpentanamido) prop-1-ynyl) phenoxy) acetate) was obtained (530 mg, 70% yield).
단계 step 2: 22: 2 -(4-(3-(-(4- (3- ( NN -- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1--One- 이닐Inil )) 페녹시Phenoxy )아세트산 (2-(4-(3-(Acetic acid (2- (4- (3- ( NN -phenylpentanamido)prop-1-ynyl)phenoxy)acetic acid)의 제조-phenylpentanamido) prop-1-ynyl) phenoxy) acetic acid)
상기 단계 1에서 수득한 에틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)페녹시)아세테이트 (530 mg, 1.35 mmol)를 에탄올 (15 ml)에 녹인 후, 상온에서 5분 동안 교반하였다. 상기 혼합물에 2N 수산화나트륨 (NaOH)(0.50 ml)을 가하고, 80oC에서 가열, 환류하여 3시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 감압 하에 농축한 뒤, 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(CH2Cl2:MeOH=100:1)로 정제하여 최종 생성물인 2-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)페녹시)아세트산(2-(4-(3-(N-phenylpentanamido)prop-1-ynyl)phenoxy)acetic acid)을 수득하였다 (246 mg, 50% 수율).The ethyl 2- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) phenoxy) acetate (530 mg, 1.35 mmol) obtained in step 1 was dissolved in ethanol (15 ml). , And stirred at room temperature for 5 minutes. To the mixture was added 2N sodium hydroxide (NaOH) (0.50 ml), heated at 80 ° C., refluxed and stirred for 3 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (CH 2 Cl 2 : MeOH = 100: 1) to give 2- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) phenoxy as a final product. ) Acetic acid (2- (4- (3- ( N- phenylpentanamido) prop-1-ynyl) phenoxy) acetic acid) was obtained (246 mg, 50% yield).
1H NMR (CDCl3, 400MHz) δ 7.47-6.78(9H, m), 4.68(2H, s), 4.58(2H, s), 2.11-2.08(2H, t), 1.58-1.53(2H, m), 1.23-1.18(2H, m), 0.81-0.78(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 7.47-6.78 (9H, m), 4.68 (2H, s), 4.58 (2H, s), 2.11-2.08 (2H, t), 1.58-1.53 (2H, m) , 1.23-1.18 (2H, m), 0.81-0.78 (3H, t).
실시예Example 30.  30. terttert -부틸 4-(5-(3-((-Butyl 4- (5- (3-(( NN -- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1-인-1-일)-1-yn-1-yl) 피콜리노일Picolinoyl )피페라진-1-카복실레이트 (Piperazine-1-carboxylate ( terttert -butyl 4-(5-(3-(N--butyl 4- (5- (3- (N- phenylpentanamidophenylpentanamido )prop-1-yn-1-yl)picolinoyl)piperazine-1-carboxylate) (LMT-834)의 제조) prop-1-yn-1-yl) picolinoyl) piperazine-1-carboxylate) (LMT-834)
상기 실시예 28의 단계 2에서 수득한 N-페닐-N-(프로프-2-이닐)펜탄아마이드와 tert-부틸 4-(5-브로모피콜리노일)피페라진-1-카복실레이트 (86 mg, 0.4 mmol)를 이용하여 실시예 28의 단계 3과 동일한 방법을 이용하여 최종 생성물인 tert-부틸 4-(5-(3-((N-페닐펜탄아미도)프로프-1-인-1-일)피콜리노일)피페라진-1-카복실레이트 (tert-butyl 4-(5-(3-(N-phenylpentanamido)prop-1-yn-1-yl)picolinoyl)piperazine-1-carboxylate)를 수득하였다 (36mg, 35% 수율). N -phenyl- N- (prop-2-ynyl) pentaneamide and tert -butyl 4- (5-bromopicolinoyl) piperazine-1-carboxylate (86 mg) obtained in step 2 of Example 28 above , 0.4 mmol) using the same method as in Step 3 of Example 28, the final product tert -butyl 4- (5- (3-(( N -phenylpentaneamido) prop-1-yn-1 -Yl) picolinoyl) piperazine-1-carboxylate (tert-butyl 4- (5- (3- (N-phenylpentanamido) prop-1-yn-1-yl) picolinoyl) piperazine-1-carboxylate) Obtained (36 mg, 35% yield).
1H NMR (400MHz, CDCl3) δ 8.43(s, 1H), 7.64(dd, 1H),7.54(dd, 1H), 7.36(dd, 3H), 7.18(m, 2H), 4.68(s, 2H), 3.69(br, 2H), 3.53-3.38(br, 6H), 2.01(m, 2H), 1.52(m, 2H), 1.39(s, 9H), 1.18(m, 2H), 0.73(t, 3H). 1 H NMR (400 MHz, CDCl 3 ) δ 8.43 (s, 1H), 7.64 (dd, 1H), 7.54 (dd, 1H), 7.36 (dd, 3H), 7.18 (m, 2H), 4.68 (s, 2H ), 3.69 (br, 2H), 3.53-3.38 (br, 6H), 2.01 (m, 2H), 1.52 (m, 2H), 1.39 (s, 9H), 1.18 (m, 2H), 0.73 (t, 3H).
실시예Example 31.  31. NN -페닐--Phenyl- NN -(3-(6-(피페라진-1--(3- (6- (piperazin-1- 카보닐Carbonyl )피리딘-3-일)Pyridin-3-yl) 프로프Prof -2-인-1-일)펜탄아마이드 (-2-yn-1-yl) pentaneamide ( NN -phenyl-N-(3-(6-(-phenyl-N- (3- (6- ( piperazinepiperazine -1-carbonyl)-1-carbonyl) pyridinpyridin -3--3- ylyl )prop-2-yn-1-yl)pentanamide) (LMT-835)의 제조) prop-2-yn-1-yl) pentanamide) (LMT-835)
상기 실시예 30에서 수득한 tert-부틸 4-(5-(3-((N-페닐펜탄아미도)프로프-1-인-1-일)피콜리노일)피페라진-1-카복실레이트 (35 mg, 0.69 mmol)를 아세토나이트릴 (15 ml)에 녹인 후, 상온에서 5분 동안 교반하였다. 상기 혼합물에 다이옥산 (dioxane)과 혼합한 하이드로클로라이드 (4N)(3.73 ml)를 가하고, 같은 온도에서 1시간 30분 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 실리카겔 컬럼 크로마토그래피 (CH2Cl2:MeOH=50:1)로 정제하여 최종 생성물인 N-페닐-N-(3-(6-(피페라진-1-카보닐)피리딘-3-일)프로프-2-인-1-일)펜탄아마이드 (N-phenyl-N-(3-(6-(piperazine-1-carbonyl)pyridin-3-yl)prop-2-yn-1-yl)pentanamide)를 수득하였다 (18 mg, 64% 수율). Tert -butyl 4- (5- (3-(( N -phenylpentaneamido) prop-1-yn-1-yl) picolinoyl) piperazine-1-carboxylate obtained in Example 30 ( 35 mg, 0.69 mmol) was dissolved in acetonitrile (15 ml) and stirred for 5 minutes at room temperature To the mixture was added hydrochloride (4N) (3.73 ml) mixed with dioxane and the same temperature The reaction mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (CH 2 Cl 2 : MeOH = 50: 1) to give N -phenyl- N- (3- as the final product. (6- (piperazin-1-carbonyl) pyridin-3-yl) prop-2-yn-1-yl) pentaneamide (N-phenyl-N- (3- (6- (piperazine-1-carbonyl ) pyridin-3-yl) prop-2-yn-1-yl) pentanamide) was obtained (18 mg, 64% yield).
1H NMR (400MHz, CDCl3) δ 8.43(s, 1H), 7.66(dd, 1H), 7.49(dd, 1H), 7.36(dd, 3H), 7.21(m, 2H), 4.67(s, 2H), 3.70(br, 2H), 3.48(br, 2H), 2.90(br, 2H), 2.81(br, 2H), 2.01(m, 2H), 1.49(m, 2H), 1.17(m, 2H), 0.73(t, 3H). 1 H NMR (400 MHz, CDCl 3 ) δ 8.43 (s, 1H), 7.66 (dd, 1H), 7.49 (dd, 1H), 7.36 (dd, 3H), 7.21 (m, 2H), 4.67 (s, 2H ), 3.70 (br, 2H), 3.48 (br, 2H), 2.90 (br, 2H), 2.81 (br, 2H), 2.01 (m, 2H), 1.49 (m, 2H), 1.17 (m, 2H) , 0.73 (t, 3 H).
실시예Example 32.  32. NN -(3-(6--(3- (6- 아이소프로필피페라진Isopropyl piperazine -1--One- 카보닐Carbonyl )피리딘-3-일)Pyridin-3-yl) 프로프Prof -2-인-1일)펜탄아마이드 (N-(3-(6-(4--2-yn-1 yl) pentaneamide (N- (3- (6- (4- isopropylpiperazineisopropylpiperazine -1-carbonyl)-1-carbonyl) pyridinpyridin -3-yl)prop-2-yn-1-yl)-N-phenylpentanamide) (LMT-836)의 제조Preparation of -3-yl) prop-2-yn-1-yl) -N-phenylpentanamide) (LMT-836)
상기 실시예 31에서 수득한 N-페닐-N-(3-(6-(피페라진-1-카보닐)피리딘-3-일)프로프-2-인-1-일)펜탄아마이드 (57.7 mg, 0.14 mmol)와 탄산수소나트륨 (27 mg, 0.316 mmol)을 얼음에서 냉각시킨 후, N,N-다이메틸포름아마이드 (DMF)(2 ml)를 넣고 1시간 동안 교반하였다. 상기 혼합물에 2-아이오도프로판 (2-iodopropane)(30 μl, 0.316 mmol)을 가하고, 얼음을 제거한 뒤, 60oC에서 가열, 환류하여 24시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 감압 하에 농축한 뒤, 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA:MeOH:TEA=12:12:1:0.1)로 정제하여 최종 생성물인 N-(3-(6-아이소프로필피페라진-1-카보닐)피리딘-3-일)프로프-2-인-1일)펜탄아마이드 (N-(3-(6-(4-isopropylpiperazine-1-carbonyl)pyridin-3-yl)prop-2-yn-1-yl)-N-phenylpentanamide)를 수득하였다 (32.3 mg, 51% 수율). N -phenyl- N- (3- (6- (piperazin-1-carbonyl) pyridin-3-yl) prop-2-yn-1-yl) pentaneamide (57.7 mg) obtained in Example 31 above. , 0.14 mmol) and sodium hydrogencarbonate (27 mg, 0.316 mmol) were cooled in ice, and N , N -dimethylformamide (DMF) (2 ml) was added thereto, followed by stirring for 1 hour. 2-iodopropane (30 μl, 0.316 mmol) was added to the mixture, ice was removed, and the mixture was heated and refluxed at 60 ° C. for 24 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA: MeOH: TEA = 12: 12: 1: 0.1) to give N- (3- (6-isopropylpiperazin-1-carbonyl) pyridine- as the final product. 3-yl) prop-2-yn-1yl) pentaneamide (N- (3- (6- (4- (4-isopropylpiperazine-1-carbonyl) pyridin-3-yl) prop-2-yn-1-yl) -N-phenylpentanamide) was obtained (32.3 mg, 51% yield).
1H NMR (400MHz, CDCl3) δ 8.44(s, 1H), 7.65(dd, 1H), 7.51(dd, 1H), 7.39(dd, 3H), 7.22(m, 2H), 4.68(s, 2H), 3.74(br, 2H), 3.52(br, 2H), 2.67(m, 1H), 2.55(br, 2H), 2.41(br, 2H), 2.02(m, 2H), 1.50(m, 2H), 1.15(m, 2H), 0.98(d, 6H), 0.74(t,3H). 1 H NMR (400 MHz, CDCl 3 ) δ 8.44 (s, 1H), 7.65 (dd, 1H), 7.51 (dd, 1H), 7.39 (dd, 3H), 7.22 (m, 2H), 4.68 (s, 2H ), 3.74 (br, 2H), 3.52 (br, 2H), 2.67 (m, 1H), 2.55 (br, 2H), 2.41 (br, 2H), 2.02 (m, 2H), 1.50 (m, 2H) , 1.15 (m, 2H), 0.98 (d, 6H), 0.74 (t, 3H).
실시예Example 33.  33. N,NN, N -- 다이에틸Diethyl -4-(3-(-4- (3- ( NN -(3--(3- 플루오로페닐Fluorophenyl )펜타아미도)Pentaamido) 프로프Prof -1-인-1-일)벤즈아마이드 (-1-yn-1-yl) benzamide ( N,NN, N -diethyl-4-(3-(N-(3--diethyl-4- (3- (N- (3- fluorophenylfluorophenyl )) pentanamidopentanamido )prop-1-prop-1- ynyn -1-yl)benzamide)의 제조(LMT-926)-1-yl) benzamide) (LMT-926)
단계 step 1 : 41: 4 -- 브로모Bromo -- N,NN, N -- 다이에틸벤즈아미드Diethylbenzamide (4- (4- bromobromo -- N,NN, N -- diethylbenzamidediethylbenzamide )의 제조Manufacturing
4-브로모벤조익산 (4-Bromobenzoic acid)(5.00g, 24.9 mmol)을 N,N-다이메틸포름아마이드(100.00 ml)에 녹인 후 다이아이소프로필아민 (13ml, 74.6 mmol)을 가하였다. 상기 혼합물에 1-하이드록시벤보트리아졸 하이드레이트(1-hydroxybenzotriazole hydrate)(7.15 mg, 37.30 mmol)와 1-에틸-3-(3-다이베틸아미노프로필)칼보다이이마이드 하이드로클로라이드 (1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) (5.04 mg, 37.30 mmol)을 가한 뒤 5분 동안 교반하였다. 상기 혼합물에 다이에틸아민 (diethylamine)(3.1ml, 37.3 mmol)을 가한 후, 상기 혼합물을 상온에서 12시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=3:1)로 정제하여 4-브로모-N,N-다이에틸벤즈아마이드 (4-bromo-N,N-diethylbenzamide)를 수득하였다 (5.70 g, 89% 수율).4-Bromobenzoic acid (4-gromobenzoic acid) (5.00 g, 24.9 mmol) was dissolved in N, N -dimethylformamide (100.00 ml) and then diisopropylamine (13 ml, 74.6 mmol) was added. To the mixture was added 1-hydroxybenzotriazole hydrate (7.15 mg, 37.30 mmol) and 1-ethyl-3- (3-dibutylaminopropyl) calvoimide hydrochloride (1-Ethyl -3- (3-dimethylaminopropyl) carbodiimide hydrochloride) (5.04 mg, 37.30 mmol) was added and stirred for 5 minutes. Diethylamine (3.1 ml, 37.3 mmol) was added to the mixture, and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 3: 1 ) to give 4-bromo-benzamide to give diethyl (4-bromo-N, N -diethylbenzamide) (5.70 g, - N, N 89% yield).
단계 2 : Step 2: N,NN, N -- 다이에틸Diethyl -4-(3--4- (3- 하이드록시프로프Hydroxyprop -1--One- 이닐Inil )) 벤즈아마이드Benzamide ( ( N,NN, N -diethyl-4-(3-hydroxyprop-1-ynyl)benzamide)의 제조Preparation of -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide)
상기 단계 1에서 수득한 4-브로모-N,N-다이에틸벤즈아마이드(5.70 mg, 22.3 mmol)와 프로파질 알코올(propagyl alcohol)(2.60 ml, 44.5 mmol)을 트리에틸아민(100.00 ml)에 녹인 후, 5분 동안 교반하였다. 상기 혼합물에 비스(트리페닐포스핀)팔라듐(II) 다이클로라이드(1.60 mg, 2.23 mmol)와 요오드화 구리(I)(1.60 mg, 2.23 mmol)를 가하고 60oC에서 가열, 환류하여 17시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 감압 하에 농축한 뒤, 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=1:1)로 정제하여 N,N-다이에틸-4-(3-하이드록시프로프-1-이닐)벤즈아마이드 (N,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzamide)를 수득하였다(5.16 mg, 99.9% 수율).4-Bromo- N, N -diethylbenzamide (5.70 mg, 22.3 mmol) and propagyl alcohol (2.60 ml, 44.5 mmol) obtained in step 1 were added to triethylamine (100.00 ml). After thawing, it was stirred for 5 minutes. Bis (triphenylphosphine) palladium (II) dichloride (1.60 mg, 2.23 mmol) and copper iodide (I) (1.60 mg, 2.23 mmol) were added to the mixture, heated at 60 ° C., refluxed and stirred for 17 hours. It was. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 1: 1) to obtain N, N -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide ( N, N- diethyl-4 -(3-hydroxyprop-1-ynyl) benzamide) was obtained (5.16 mg, 99.9% yield).
단계 step 3 : 33: 3 -(4-(-(4-( N,NN, N -- 다이에틸카르바모일Diethylcarbamoyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil 메탄설포네이트Methanesulfonate (3-(4-(diethylcarbamoyl)phenyl)prop-2-yn-1-yl methanesulfonate)의 제조 Preparation of (3- (4- (diethylcarbamoyl) phenyl) prop-2-yn-1-yl methanesulfonate)
N,N-다이에틸-4-(3-하이드록시프로프-1-이닐)벤즈아마이드 (5.16 mg, 22.3 mmol)을 다이클로로메탄(100 ml)에 녹인 후 얼음에서 냉각시켰다. 상기 혼합물에 트리에틸아민(4.80 ml, 34.4 mmol)을 가한 뒤 5분 동안 교반하였다. 같은 온도에서 메탄설포닐클로라이드(Methanesulfonyl chloride)(1.95 ml, 25.2 mmol)를 가하고, 얼음을 제거한 뒤, 상기 혼합물을 상온에서 30분 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=2:1)로 정제하여 3-(4-(N,N-다이에틸카르바모일)페닐)프로프-2-이닐 메탄설포네이트 (3-(4-(diethylcarbamoyl)phenyl)prop-2-yn-1-yl methanesulfonate)를 수득하였다(4.2 mg, 59% 수율). N, N -diethyl-4- (3-hydroxyprop-1-ynyl) benzamide (5.16 mg, 22.3 mmol) was dissolved in dichloromethane (100 ml) and cooled on ice. Triethylamine (4.80 ml, 34.4 mmol) was added to the mixture, which was then stirred for 5 minutes. Methanesulfonyl chloride (1.95 ml, 25.2 mmol) was added at the same temperature, ice was removed, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 2: 1) to give 3- (4- (N, N-diethylcarbamoyl) phenyl) prop-2-ynyl methanesulfonate (3- ( 4- (diethylcarbamoyl) phenyl) prop-2-yn-1-yl methanesulfonate) was obtained (4.2 mg, 59% yield).
단계 4 : Step 4: N,NN, N -- 다이에틸Diethyl -4-(3-(N-(3--4- (3- (N- (3- 플루오로페닐Fluorophenyl )펜타아미도)Pentaamido) 프로프Prof -1-인-1-일)벤즈아마이드 (-1-yn-1-yl) benzamide ( N,NN, N -diethyl-4-(3-(-diethyl-4- (3- ( NN -(3--(3- fluorophenylfluorophenyl )) pentanamidopentanamido )prop-1-prop-1- ynyn -1-yl)benzamide)의 제조-1-yl) benzamide)
N-(3-플루오로페닐)펜탄아마이드 (200mg, 1.02 mmol)을 테트라하이드로퓨란(10.00 ml)에 녹인 후 얼음에서 냉각시켰다. 상기 혼합물에 수소화나트륨(73 mg, 3.06 mmol)을 가한 뒤 1시간 동안 교반하였다. 같은 온도에서 상기 단계 3에서 수득한 3-(4-(N,N-다이에틸카르바모일)페닐)프로프-2-이닐 메탄설포네이트 (475mg, 1.54 mmol)을 가하고, 상기 혼합물을 상온에서 4시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=2:1)로 정제하여 N,N-다이에틸-4-(3-(N-(3-플루오로페닐)펜타아미도)프로프-1-인-1-일)벤즈아마이드(N,N-diethyl-4-(3-(N-(3-fluorophenyl)pentanamido)prop-1-yn-1-yl)benzamide)를 수득하였다(291.7 mg, 70% 수율). N- (3-fluorophenyl) pentaneamide (200 mg, 1.02 mmol) was dissolved in tetrahydrofuran (10.00 ml) and cooled on ice. Sodium hydride (73 mg, 3.06 mmol) was added to the mixture, followed by stirring for 1 hour. At the same temperature, 3- (4- ( N, N -diethylcarbamoyl) phenyl) prop-2-ynyl methanesulfonate (475 mg, 1.54 mmol) obtained in step 3 was added thereto, and the mixture was allowed to stand at room temperature. Stir for 4 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 2: 1) to give N, N -diethyl-4- (3- (N- (3-fluorophenyl) pentamido) prop-1- In-1-yl) benzamide ( N, N- diethyl-4- (3- ( N- (3-fluorophenyl) pentanamido) prop-1-yn-1-yl) benzamide) was obtained (291.7 mg, 70 % Yield).
1H NMR (CDCl3, 500MHz) δ 7.43 (1H, t, J = 7.5 Hz and 15.0 Hz, aromatic), 7.33 (4H, m, aromatic), 7.09 (3H, m, aromatic), 4.71 (2H, s, CH2), 3.53 (2H, s, CH2), 3.23 (2H, s, CH2), 2.10 (2H, m, CH2), 1.59 (2H, m, CH2), 1.24 (2H, m, CH2), 1.10 (6H, m, (CH3)2), 0.83 (3H, t, J = 7.5 Hz and 15.0 Hz, CH3).1 H NMR (CDCl 3 , 500 MHz) δ 7.43 (1H, t, J = 7.5 Hz and 15.0 Hz, aromatic), 7.33 (4H, m, aromatic), 7.09 (3H, m, aromatic), 4.71 (2H, s, CH 2 ), 3.53 (2H, s, CH 2 ), 3.23 (2H, s, CH 2 ), 2.10 (2H, m, CH 2 ), 1.59 (2H, m, CH 2 ), 1.24 (2H, m, CH 2 ), 1.10 (6H, m, (CH 3 ) 2 ), 0.83 (3H, t, J = 7.5 Hz and 15.0 Hz, CH 3 ).
실시예Example 34.  34. N,NN, N -다이에틸-4-(3-(-Diethyl-4- (3- ( NN -(4-플루오로페닐)펜타아미도)프로프-1-인-1-일)벤즈아마이드(-(4-fluorophenyl) pentamido) prop-1-yn-1-yl) benzamide ( N,NN, N -diethyl-4-(3-(-diethyl-4- (3- ( NN -(4-fluorophenyl)pentanamido)prop-1-yn-1-yl)benzamide)의 제조(LMT-927)-(4-fluorophenyl) pentanamido) prop-1-yn-1-yl) benzamide) (LMT-927)
N-(4-플루오로페닐)펜탄아마이드 (200mg, 1.02 mmol)과 3-(4-(N,N-다이에틸카르바모일)페닐)프로프-2-이닐 메탄설포네이트 (3-(4-(diethylcarbamoyl)phenyl)prop-2-yn-1-yl methanesulfonate)을 이용하여 상기 실시예 33의 단계 4의 방법을 이용하여 N,N-다이에틸-4-(3-(N-(4-플루오로페닐)펜타아미도)프로프-1-인-1-일)벤즈아마이드(N,N-diethyl-4-(3-(N-(4-fluorophenyl)pentanamido)prop-1-yn-1-yl)benzamide)를 수득하였다(291.7 mg, 70% 수율). N- (4-fluorophenyl) pentaneamide (200 mg, 1.02 mmol) and 3- (4- ( N, N -diethylcarbamoyl) phenyl) prop-2-ynyl methanesulfonate (3- (4 -(diethylcarbamoyl) phenyl) prop-2-yn-1-yl methanesulfonate) using N, N-diethyl-4- (3- (N- (4- Fluophenyl) pentaamido) prop-1-yn-1-yl) benzamide ( N, N -diethyl-4- (3- ( N- (4-fluorophenyl) pentanamido) prop-1-yn-1 -yl) benzamide) was obtained (291.7 mg, 70% yield).
1H NMR (CDCl3, 500MHz) δ 7.31 (2H, d, J = 3.5 Hz, aromatic), 7.29 (4H, d, J = 3.0 Hz, aromatic), 7.15 (2H, t, J = 8.5 Hz and 17.0 Hz, aromatic), 4.71 (2H, s, CH2), 3.53 (2H, s, CH2), 3.23 (2H, s, CH2), 2.06 (2H, t, J = 7.5 Hz and 15.0 Hz, CH2), 1.57 (2H, m, CH2), 1.22 (2H, m, CH2), 1.10 (6H, s, (CH3)2), 0.82 (3H, t, J = 7.5 Hz and 15.0 Hz, CH3). 1 H NMR (CDCl 3 , 500 MHz) δ 7.31 (2H, d, J = 3.5 Hz, aromatic), 7.29 (4H, d, J = 3.0 Hz, aromatic), 7.15 (2H, t, J = 8.5 Hz and 17.0 Hz, aromatic), 4.71 (2H, s, CH 2 ), 3.53 (2H, s, CH 2 ), 3.23 (2H, s, CH 2 ), 2.06 (2H, t, J = 7.5 Hz and 15.0 Hz, CH 2 ), 1.57 (2H, m, CH 2 ), 1.22 (2H, m, CH 2 ), 1.10 (6H, s, (CH 3 ) 2 ), 0.82 (3H, t, J = 7.5 Hz and 15.0 Hz, CH 3 ).
실시예Example 35.  35. NN -(3-(4-(-(3- (4- ( N,NN, N -- 다이에틸설파모일Diethylsulfamoyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜타아마이드Phenylpentamide ( ( NN -(3-(4-(-(3- (4- ( N,NN, N -- diethylsulfamoyldiethylsulfamoyl )phenyl)prop-2-) phenyl) prop-2- ynylynyl )-)- NN -phenylpentanamide)의 제조(LMT-946)-phenylpentanamide) (LMT-946)
단계 step 1 : 41: 4 -- 브로모Bromo -- N,NN, N -- 다이에틸벤젠설폰아마이드Diethylbenzenesulfonamide (4- (4- bromobromo -- N,NN, N -diethylbenzenesulfonamide)의 제조-diethylbenzenesulfonamide)
4-브로모벤젠설포닐 클로라이드(4-Bromobenzenesulfonyl chloride)(1.00g, 3.91 mmol)을 다이클로로메탄(30.00 ml)에 녹인 후 얼음에서 냉각시켰다. 상기 혼합물에 다이에틸아민(1.19 ml, 11.54 mmol)을 가한 뒤 5분 동안 교반하고, 얼음을 제거한 뒤, 상기 혼합물을 상온에서 12시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=3:1)로 정제하여 4-브로모-N,N-다이에틸벤젠설폰아마이드 (4-bromo-N,N-diethylbenzenesulfonamide)를 수득하였다 (1.10 g, 96% 수율).4-Bromobenzenesulfonyl chloride (1.00 g, 3.91 mmol) was dissolved in dichloromethane (30.00 ml) and cooled on ice. Diethylamine (1.19 ml, 11.54 mmol) was added to the mixture, followed by stirring for 5 minutes. After removing ice, the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 3: 1) to give 4-bromo- N, N -diethylbenzenesulfonamide (4-bromo- N, N- diethylbenzenesulfonamide) (1.10 g , 96% yield).
1H NMR (CDCl3, 500MHz) δ 7.67 (4H, m), 3.23 (4H, q, J = 7.0 Hz), 1.13 (6H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.67 (4H, m), 3.23 (4H, q, J = 7.0 Hz), 1.13 (6H, t).
단계 2 : Step 2: N,NN, N -- 다이에틸Diethyl -4-(3--4- (3- 하이드록시프로프Hydroxyprop -1--One- 이닐Inil )) 벤젠설폰아마이드Benzenesulfonamide ( ( N,NN, N -diethyl-4-(3-hydroxyprop-1-ynyl)benzenesulfonamide)의 제조Preparation of -diethyl-4- (3-hydroxyprop-1-ynyl) benzenesulfonamide)
상기 단계 1에서 수득한 4-브로모-N,N-다이에틸벤젠설폰아마이드(500.00 mg, 1.71 mmol)와 프로파질 알코올(propagyl alcohol)(0.20 ml, 3.42 mmol)을 트리에틸아민(10.00 ml)에 녹인 후, 5분 동안 교반하였다. 상기 혼합물에 비스(트리페닐포스핀)팔라듐(II) 다이클로라이드(119.32 mg, 0.17 mmol)와 요오드화 구리(I)(32.37 mg, 0.17 mmol)를 가하고 60oC에서 가열, 환류하여 17시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 감압 하에 농축한 뒤, 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=1:1)로 정제하여 N,N-다이에틸-4-(3-하이드록시프로프-1-이닐)벤젠설폰아마이드 (N,N-diethyl-4-(3-hydroxyprop-1-ynyl)benzenesulfonamide)를 수득하였다(286.00 mg, 63% 수율).4-Bromo- N, N -diethylbenzenesulfonamide (500.00 mg, 1.71 mmol) and propagyl alcohol (0.20 ml, 3.42 mmol) obtained in step 1 were added with triethylamine (10.00 ml). It was dissolved in and stirred for 5 minutes. Bis (triphenylphosphine) palladium (II) dichloride (119.32 mg, 0.17 mmol) and copper iodide (I) (32.37 mg, 0.17 mmol) were added to the mixture, which was heated and refluxed at 60 ° C. for 17 hours. It was. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 1: 1) to obtain N, N -diethyl-4- (3-hydroxyprop-1-ynyl) benzenesulfonamide ( N, N -diethyl- 4- (3-hydroxyprop-1-ynyl) benzenesulfonamide) was obtained (286.00 mg, 63% yield).
1H NMR (CDCl3, 500MHz) δ 7.66 (2H, d, J = 8.0 Hz), 7.42 (2H, d, J = 8.0 Hz), 4.44 (2H, s), 3.16 (4H, q, J = 7.0 Hz), 1.05 (6H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.66 (2H, d, J = 8.0 Hz), 7.42 (2H, d, J = 8.0 Hz), 4.44 (2H, s), 3.16 (4H, q, J = 7.0 Hz), 1.05 (6H, t).
단계 step 3 : 33: 3 -(4-(-(4-( N,NN, N -- 다이에틸설파모일Diethylsulfamoyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil 메탄설포네이트Methanesulfonate (3-(4-( (3- (4- ( N,NN, N -diethylsulfamoyl)phenyl)prop-2-ynyl methanesulfonate)의 제조-diethylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate)
상기 단계 2에서 수득한 N,N-다이에틸-4-(3-하이드록시프로프-1-이닐)벤젠설폰아마이드 (273.00 mg, 1.02 mmol)을 다이클로로메탄(10 ml)에 녹인 후 얼음에서 냉각시켰다. 상기 혼합물에 트리에틸아민(0.21 ml, 1.53 mmol)을 가한 뒤 5분 동안 교반하였다. 같은 온도에서 메탄설포닐클로라이드(Methanesulfonyl chloride)(0.09 ml, 1.12 mmol)를 가하고, 얼음을 제거한 뒤, 상기 혼합물을 상온에서 30분 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산 마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=2:1)로 정제하여 3-(4-(N,N-다이에틸설파모일)페닐)프로프-2-이닐 메탄설포네이트 (3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynyl methanesulfonate)를 수득하였다(285.00 mg, 80% 수율). N, N -diethyl-4- (3-hydroxyprop-1-ynyl) benzenesulfonamide (273.00 mg, 1.02 mmol) obtained in step 2 was dissolved in dichloromethane (10 ml), and then dried on ice. Cooled. Triethylamine (0.21 ml, 1.53 mmol) was added to the mixture, which was then stirred for 5 minutes. Methanesulfonyl chloride (0.09 ml, 1.12 mmol) was added at the same temperature, ice was removed, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 2: 1) to give 3- (4- ( N, N -diethylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate (3- (4 -( N, N- diethylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate) was obtained (285.00 mg, 80% yield).
1H NMR (CDCl3, 500MHz) δ 7.73 (2H, d, J = 8.5 Hz), 7.53 (2H, d, J = 8.5 Hz), 5.05 (2H, s), 3.19 (4H, q, J = 7.0 Hz), 3.12 (3H, s), 1.07 (6H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.73 (2H, d, J = 8.5 Hz), 7.53 (2H, d, J = 8.5 Hz), 5.05 (2H, s), 3.19 (4H, q, J = 7.0 Hz), 3.12 (3H, s), 1.07 (6H, t).
단계 step 4 : 44: 4 -(3--(3- 브로모프로프Bromoprop -1--One- 이닐Inil )-)- N,NN, N -- 다이에틸벤젠설폰아마이드Diethylbenzenesulfonamide (4-(3-bromoprop-1-ynyl)- (4- (3-bromoprop-1-ynyl)- N,NN, N -diethylbenzenesulfonamide)의 제조-diethylbenzenesulfonamide)
상기 단계 3에서 수득한 3-(4-(N,N-다이에틸설파모일)페닐)프로프-2-이닐 메탄설포네이트 (260.00 mg, 0.75 mmol)을 테트라하이드로퓨란(Tetrahydrofuran)(20.00 ml)에 녹인 후 얼음에서 냉각시켰다. 같은 온도에서 상기 혼합물에 리튬브로마이드(Lithium bromide)(196.28 mg, 2.26 mmol)을 가한 뒤 4시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산 마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 필터로 정제하여 4-(3-브로모프로프-1-이닐)-N,N-다이에틸벤젠설폰아마이드 (4-(3-bromoprop-1-ynyl)-N,N-diethylbenzenesulfonamide)를 수득하였다.(240.00 mg, 97 %)3- (4- ( N, N -diethylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate (260.00 mg, 0.75 mmol) obtained in step 3 was added to tetrahydrofuran (20.00 ml). It was dissolved in and cooled on ice. Lithium bromide (196.28 mg, 2.26 mmol) was added to the mixture at the same temperature, followed by stirring for 4 hours. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered, and concentrated under reduced pressure. The concentrate was purified by a filter to afford 4- (3-bromoprop-1-ynyl) -N, N -diethylbenzenesulfonamide (4- (3-bromoprop-1-ynyl) -N, N -diethylbenzenesulfonamide). Obtained (240.00 mg, 97%)
단계 5 : Step 5: N,NN, N -- 다이에틸Diethyl -4-(3-(-4- (3- ( 페닐아미노Phenylamino )) 프로프Prof -1--One- 이닐Inil )) 벤젠설폰아마이드Benzenesulfonamide ( ( N,NN, N -diethyl-4-(3-(phenylamino)prop-1-ynyl)benzenesulfonamide)의 제조Preparation of -diethyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide)
상기 단계 4에서 수득한 4-(3-브로모프로프-1-이닐)-N,N-다이에틸벤젠설폰아마이드(248.00mg, 0.75 mmol)와 포타시움카보네이트(Potassium carbonate)(93.98 mg, 0.68 mmol)를 아세토나이트릴(Acetonitrile)(15.00 ml)에 녹여 30분 동안 교반하였다. 상기 혼합물에 아닐린(aniline)(0.06ml, 0.68 mmol)을 가한 뒤 상온에서 9시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=2:1)로 정제하여 N,N-다이에틸-4-(3-(페닐아미노)프로프-1-이닐)벤젠설폰아마이드 (N,N-diethyl-4-(3-(phenylamino)prop-1-ynyl)benzenesulfonamide)를 수득하였다(190.00 mg, 81% 수율).4- (3-bromoprop-1-ynyl) -N, N -diethylbenzenesulfonamide (248.00 mg, 0.75 mmol) and Potassium carbonate (93.98 mg, 0.68 mmol) obtained in step 4 above. Was dissolved in acetonitrile (15.00 ml) and stirred for 30 minutes. Aniline (0.06ml, 0.68 mmol) was added to the mixture, followed by stirring at room temperature for 9 hours. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 2: 1) to give N, N -diethyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide ( N, N − diethyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide) was obtained (190.00 mg, 81% yield).
1H NMR (CDCl3, 500MHz) δ 7.71 (2H, d, J = 8.5 Hz), 7.46 (2H, d, J = 8.5 Hz), 7.23 (2H, t), 6.80 (1H, t), 6.73(2H, d, J = 7.5 Hz), 4.15 (2H, s), 3.21 (4H, m), 1.10 (6H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.71 (2H, d, J = 8.5 Hz), 7.46 (2H, d, J = 8.5 Hz), 7.23 (2H, t), 6.80 (1H, t), 6.73 ( 2H, d, J = 7.5 Hz), 4.15 (2H, s), 3.21 (4H, m), 1.10 (6H, t).
단계 6 : Step 6: NN -(3-(4-(-(3- (4- ( N,NN, N -- 다이에틸설파모일Diethylsulfamoyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜타아마이드Phenylpentamide ( ( NN -(3-(4-(-(3- (4- ( N,NN, N -- diethylsulfamoyldiethylsulfamoyl )phenyl)prop-2-) phenyl) prop-2- ynylynyl )-)- NN -phenylpentanamide)의 제조-phenylpentanamide)
상기 단계 5에서 수득한 N,N-다이에틸-4-(3-(페닐아미노)프로프-1-이닐)벤젠설폰아마이드 (174.00mg, 0.51 mmol)을 다이클로로메탄(15.00 ml)에 녹인 후 얼음에서 냉각시켰다. 상기 혼합물에 트리에틸아민(0.14 ml, 1.02 mmol)을 가한 뒤 5분 동안 교반하였다. 같은 온도에서 발레로일 클로라이드(valeroyl chloride)(0.06 ml, 0.53 mmol)를 가하고, 얼음을 제거한 뒤, 상기 혼합물을 상온에서 4시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=2:1)로 정제하여 N-(3-(4-(N,N-다이에틸설파모일)페닐)프로프-2-이닐)-N-페닐펜타아마이드 (N-(3-(4-(N,N-diethylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다(153.00 mg, 70% 수율). N, N -diethyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide (174.00 mg, 0.51 mmol) obtained in step 5 was dissolved in dichloromethane (15.00 ml). Cooled on ice. Triethylamine (0.14 ml, 1.02 mmol) was added to the mixture, which was then stirred for 5 minutes. Valeroyl chloride (0.06 ml, 0.53 mmol) was added at the same temperature, ice was removed, and the mixture was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 2: 1) to give N- (3- (4- ( N, N -diethylsulfamoyl) phenyl) prop-2-ynyl) -N -phenyl penta amide (N - (3- (4- ( N, N -diethylsulfamoyl) phenyl) prop-2-ynyl) - N -phenylpentanamide) was obtained (153.00 mg, 70% yield).
1H NMR (CDCl3, 500MHz) δ 7.74 (2H, d, J = 8.0 Hz), 7.51 (4H, m), 7.43 (3H, m), 4.73 (2H, s), 3.21 (4H, s), 2.11 (2H, t), 1.52 (2H, m), 1.21 (2H, m), 0.10 (6H, t), 0.80 (3H, t). 1 H NMR (CDCl 3 , 500 MHz) δ 7.74 (2H, d, J = 8.0 Hz), 7.51 (4H, m), 7.43 (3H, m), 4.73 (2H, s), 3.21 (4H, s), 2.11 (2H, t), 1.52 (2H, m), 1.21 (2H, m), 0.10 (6H, t), 0.80 (3H, t).
실시예Example 36.  36. NN -(3-(4-(-(3- (4- ( NN -- 아이소프로필설파모일Isopropyl sulfamoyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜타아마이드Phenylpentamide ( ( NN -(3-(4-(-(3- (4- ( NN -- isopropylsulfamoylisopropylsulfamoyl )phenyl)prop-2-) phenyl) prop-2- ynylynyl )-)- NN -phenylpentanamide)의 제조(LMT-947)-phenylpentanamide) (LMT-947)
단계 step 1 : 41: 4 -- 브로모Bromo -- NN -- 아이소프로필벤젠설폰아마이드Isopropylbenzenesulfonamide (4- (4- bromobromo -- NN -isopropylbenzenesulfonamide)의 제조-isopropylbenzenesulfonamide)
4-브로모벤젠설포닐 클로라이드 (4-Bromobenzenesulfonyl chloride)(1.00g, 3.91 mmol)을 다이클로로메탄(10.00 ml)에 녹인 후 상기 혼합물에 아이소프로필아민(0.40 ml, 4.69 mmol)과 피리딘(0.41 ml, 5.09 mmol)을 가한 뒤 상온에서 2시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=3:1)로 정제하여 4-브로모-N-아이소프로필벤젠설폰아마이드 (4-bromo-N-isopropylbenzenesulfonamide)를 수득하였다. (730.00 mg, 67% 수율).4-Bromobenzenesulfonyl chloride (1.00 g, 3.91 mmol) was dissolved in dichloromethane (10.00 ml) and isopropylamine (0.40 ml, 4.69 mmol) and pyridine (0.41 ml) were added to the mixture. , 5.09 mmol) was added, followed by stirring at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 3: 1) to give 4-bromo- N -isopropylbenzenesulfonamide (4-bromo- N- isopropylbenzenesulfonamide). (730.00 mg, 67% yield).
1H NMR (CDCl3, 500 MHz) δ 7.76-7.64 (4H, m), 3.47 (1H, m), 1.09 (6H, d, J = 7.0 Hz). 1 H NMR (CDCl 3 , 500 MHz) δ 7.76-7.64 (4H, m), 3.47 (1H, m), 1.09 (6H, d, J = 7.0 Hz).
단계 step 2 : 42: 4 -(3--(3- 하이드록시프로프Hydroxyprop -1--One- 이닐Inil )-)- NN -- 아이소프로필벤젠설폰아마이드Isopropylbenzenesulfonamide (4-(3-hydroxyprop-1-ynyl)- (4- (3-hydroxyprop-1-ynyl)- NN -isopropylbenzenesulfonamide)의 제조-isopropylbenzenesulfonamide)
상기 단계 1에서 수득한 4-브로모-N-아이소프로필벤젠설폰아마이드(500.00 mg, 1.80 mmol)와 프로파질 알코올(propagyl alcohol)(0.21 ml, 3.59 mmol)을 이용하여 상기 실시예 35의 단계 2와 동일한 방법으로 4-(3-하이드록시프로프-1-이닐)-N-아이소프로필벤젠설폰아마이드 (4-(3-hydroxyprop-1-ynyl)-N-isopropylbenzenesulfonamide)를 수득하였다(420.00 mg, 92% 수율). Step 2 of Example 35 using 4-bromo- N -isopropylbenzenesulfonamide (500.00 mg, 1.80 mmol) and propagyl alcohol (0.21 ml, 3.59 mmol) obtained in step 1 above. In the same manner as in 4- (3-hydroxyprop-1-ynyl) -N -isopropylbenzenesulfonamide (4- (3-hydroxyprop-1-ynyl) -N- isopropylbenzenesulfonamide) was obtained (420.00 mg, 92% yield).
1H NMR (CDCl3, 500MHz) δ 7.81 (2H, d, J = 8.0 Hz), 7.49 (2H, d, J = 8.5 Hz), 4.51 (2H, s), 3.44 (1H, m), 1.06 (6H, d, J = 6.5 Hz). 1 H NMR (CDCl 3 , 500 MHz) δ 7.81 (2H, d, J = 8.0 Hz), 7.49 (2H, d, J = 8.5 Hz), 4.51 (2H, s), 3.44 (1H, m), 1.06 ( 6H, d, J = 6.5 Hz).
단계 step 3 : 33: 3 -(4-(-(4-( NN -- 아이소프로필설파모일Isopropyl sulfamoyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil 메탄설포네이트Methanesulfonate (3-(4-( (3- (4- ( NN -isopropylsulfamoyl)phenyl)prop-2-ynyl methanesulfonate)의 제조-isopropylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate)
상기 단계 2에서 수득한 4-(3-하이드록시프로프-1-이닐)-N-아이소프로필벤젠설폰아마이드 (410.00 mg, 1.62 mmol)와 메탄설포닐클로라이드(Methanesulfonyl chloride)(0.14 ml, 1.78 mmol)을 이용하여 상기 실시예 35의 단계 3와 동일한 방법으로 3-(4-(N-아이소프로필설파모일)페닐)프로프-2-이닐 메탄설포네이트 (3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynyl methanesulfonate)를 수득하였다( 330.00 mg, 61 % 수율).4- (3-hydroxyprop-1-ynyl) -N -isopropylbenzenesulfonamide (410.00 mg, 1.62 mmol) and methanesulfonyl chloride (0.14 ml, 1.78 mmol) obtained in step 2; In the same manner as in step 3 of Example 35, using 3- (4- ( N -isopropylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate (3- (4- ( N- isopropylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate) was obtained (330.00 mg, 61% yield).
1H NMR (CDCl3, 500MHz) δ 7.86 (2H, d, J = 8.5 Hz), 7.59 (2H, d, J = 8.5 Hz), 5.10 (2H, s), 3.47 (1H, m) 3.17 (3H, s), 1.08 (6H, d, J = 6.5 Hz). 1 H NMR (CDCl 3 , 500 MHz) δ 7.86 (2H, d, J = 8.5 Hz), 7.59 (2H, d, J = 8.5 Hz), 5.10 (2H, s), 3.47 (1H, m) 3.17 (3H , s), 1.08 (6H, d, J = 6.5 Hz).
단계 step 4 : 44: 4 -(3--(3- 브로모프로프Bromoprop -1--One- 이닐Inil )-)- NN -- 아이소프로필벤젠설폰아마이드Isopropylbenzenesulfonamide (4-(3-bromoprop-1-ynyl)- (4- (3-bromoprop-1-ynyl)- NN -isopropylbenzenesulfonamide)의 제조-isopropylbenzenesulfonamide)
상기 단계 3에서 수득한 3-(4-(N-아이소프로필설파모일)페닐)프로프-2-이닐 메탄설포네이트 (210.00 mg, 0.63 mmol)와 리튬브로마이드(Lithium bromide)(165.00 mg, 1.90 mmol)을 이용하여 상기 실시예 35의 단계 4와 동일한 방법으로 4-(3-브로모프로프-1-이닐)-N-아이소프로필벤젠설폰아마이드 (4-(3-bromoprop-1-ynyl)-N-isopropylbenzenesulfonamide)를 수득하였다(190.00 mg, 95% 수율).3- (4- ( N -isopropylsulfamoyl) phenyl) prop-2-ynyl methanesulfonate (210.00 mg, 0.63 mmol) and lithium bromide (165.00 mg, 1.90 mmol) obtained in step 3 above. ) to the same manner as in step 4 of example 35 using 4- (3-bromo-1-ynyl-morph rope) - N - isopropyl-benzenesulfonamide (4- (3-bromoprop-1 -ynyl) -N -isopropylbenzenesulfonamide) was obtained (190.00 mg, 95% yield).
단계 5 : Step 5: NN -- 아이소프로필Isopropyl -4-(3-(-4- (3- ( 페닐아미노Phenylamino )) 프로프Prof -1--One- 이닐Inil )) 벤젠설폰아마이드Benzenesulfonamide ( ( NN -isopropyl-4-(3-(phenylamino)prop-1-ynyl)benzenesulfonamide)의 제조Preparation of -isopropyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide)
상기 단계 4에서 수득한 4-(3-브로모프로프-1-이닐)-N-아이소프로필벤젠설폰아마이드 (199.00 mg, 0.63 mmol)와 아닐린(aniline)(0.05 ml, 0.57 mmol)을 이용하여 상기 실시예 35의 단계 5와 동일한 방법으로 N-아이소프로필-4-(3-(페닐아미노)프로프-1-이닐)벤젠설폰아마이드 (N-isopropyl-4-(3-(phenylamino)prop-1-ynyl)benzenesulfonamide)를 수득하였다(157 mg, 83 % 수율).4- (3-bromoprop-1-ynyl) -N -isopropylbenzenesulfonamide (199.00 mg, 0.63 mmol) and aniline (0.05 ml, 0.57 mmol) obtained in step 4 above were used. N -isopropyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide in the same manner as in Step 5 of Example 35 ( N- isopropyl-4- (3- (phenylamino) prop-1 -ynyl) benzenesulfonamide) was obtained (157 mg, 83% yield).
1H NMR (CDCl3, 500MHz) δ 7.80 (2H, d, J = 9.0 Hz), 7.50 (2H, d, J = 8.5 Hz), 7.25 (2H, m), 6.82 (1H, t), 6.76 (2H, d, J = 8.0 Hz), 4.64 (1H, d, J = 8.0 Hz), 4.19 (2H, s), 3.45 (1H, m), 1.06 (6H, d, J = 6.0 Hz). 1 H NMR (CDCl 3 , 500 MHz) δ 7.80 (2H, d, J = 9.0 Hz), 7.50 (2H, d, J = 8.5 Hz), 7.25 (2H, m), 6.82 (1H, t), 6.76 ( 2H, d, J = 8.0 Hz), 4.64 (1H, d, J = 8.0 Hz), 4.19 (2H, s), 3.45 (1H, m), 1.06 (6H, d, J = 6.0 Hz).
단계 6 : Step 6: NN -(3-(4-(-(3- (4- ( NN -- 아이소프로필설파모일Isopropyl sulfamoyl )페닐)) Phenyl) 프로프Prof -2--2- 이닐Inil )-)- NN -- 페닐펜탄아마이드Phenylpentaneamide ( ( NN -(3-(4-(-(3- (4- ( NN -- isopropylsulfamoylisopropylsulfamoyl )phenyl)prop-2-) phenyl) prop-2- ynylynyl )-)- NN -phenylpentanamide)의 제조-phenylpentanamide)
상기 단계 2에서 수득한 N-아이소프로필-4-(3-(페닐아미노)프로프-1-이닐)벤젠설폰아마이드(150.00 mg, 0.46 mmol)와 발레로일 클로라이드(valeroyl chloride)(0.06 ml, 0.48 mmol)을 이용하여 상기 실시예 35의 단계 6와 동일한 방법으로 N-(3-(4-(N-아이소프로필설파모일)페닐)프로프-2-이닐)-N-페닐펜탄아마이드 (N-(3-(4-(N-isopropylsulfamoyl)phenyl)prop-2-ynyl)-N-phenylpentanamide)를 수득하였다.(94.00 mg, 49.5 % 수율). N -isopropyl-4- (3- (phenylamino) prop-1-ynyl) benzenesulfonamide (150.00 mg, 0.46 mmol) and valeroyl chloride (0.06 ml, obtained in step 2) 0.48 mmol) to the same method as in step 6 of example 35 using the N - (3- (4- (N - isopropyl-sulfamoyl) phenyl) prop-2-ynyl) - N - phenyl pentane amide (N -(3- (4- ( N- isopropylsulfamoyl) phenyl) prop-2-ynyl) -N- phenylpentanamide) was obtained (94.00 mg, 49.5% yield).
1H NMR (CDCl3, 500MHz) δ 7.78 (2H, d, J = 8.5 Hz), 7.51-7.37 (7H, m), 4.72 (2H, s), 3.33 (1H, m), 2.10 (2H, t), 1.51 (2H, m), 1.19 (2H, m), 0.99 (6H, d, J = 6.5 Hz), 0.78 (3H, t) 1 H NMR (CDCl 3 , 500 MHz) δ 7.78 (2H, d, J = 8.5 Hz), 7.51-7.37 (7H, m), 4.72 (2H, s), 3.33 (1H, m), 2.10 (2H, t ), 1.51 (2H, m), 1.19 (2H, m), 0.99 (6H, d, J = 6.5 Hz), 0.78 (3H, t)
실시예Example 37.  37. terttert -부틸 4--Butyl 4- (3-((3- ( NN -페닐펜탄아미도)프로프Phenylpentane amido) prop -1-인-1-일)-1-yn-1-yl) 벤조에이Benzoa 트(tert-butyl 4-(3-(Tert-butyl 4- (3- ( NN -- phenylpentanamidophenylpentanamido )prop-1-prop-1- ynyn -1--One- ylyl )benzoate)의 제조() benzoate) LMTLMT -1012)-1012)
단계 1 : Step 1: NN -페닐펜탄아마이드(N-phenylpentanamide)의 제조Preparation of N-phenylpentanamide
아닐린(aniline)(10.00 ml, 107.40 mmol)을 다이클로로메탄(150 ml)에 녹인 후 얼음에서 냉각시켰다. 상기 혼합물에 트리에틸아민(30.00 ml, 214.80 mmol)을 가한 뒤 5분 동안 교반하였다. 같은 온도에서 발레로일 클로라이드(valeroyl chloride)(16.00 ml, 128.90 mmol)를 가하고, 얼음을 제거한 뒤, 상기 혼합물을 상온에서 2시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=10:1)로 정제하여 N-페닐펜탄아마이드(N-phenylpentanamide)를 수득하였다(19.1 g, 99.9% 수율).Aniline (10.00 ml, 107.40 mmol) was dissolved in dichloromethane (150 ml) and then cooled on ice. Triethylamine (30.00 ml, 214.80 mmol) was added to the mixture and stirred for 5 minutes. Valeroyl chloride (16.00 ml, 128.90 mmol) was added at the same temperature, ice was removed, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the obtained residue was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. To give a phenyl-pentane amide (N -phenylpentanamide) (19.1 g, 99.9% yield) to give the N in the concentrate was purified by silica gel column chromatography (1 Hex:: EA = 10 ).
1H NMR (CDCl3, 500MHz) δ 7.55 (2H, d, J = 8.0 Hz, aromatic), 7.29 (2H, t, J = 7.5 Hz and 15.0 Hz, aromatic), 7.09 (1H, t, J = 7.0 Hz and 14.5 Hz, aromatic), 2.35 (2H, t, J = 8.0 Hz and 15.5 Hz, CH2), 1.70 (2H, m, CH2), 1.37 (2H, m, CH2), 0.92 (3H, t, J = 7.0 Hz and 14.5 Hz, CH3). 1 H NMR (CDCl 3 , 500 MHz) δ 7.55 (2H, d, J = 8.0 Hz, aromatic), 7.29 (2H, t, J = 7.5 Hz and 15.0 Hz, aromatic), 7.09 (1H, t, J = 7.0 Hz and 14.5 Hz, aromatic), 2.35 (2H, t, J = 8.0 Hz and 15.5 Hz, CH 2 ), 1.70 (2H, m, CH 2 ), 1.37 (2H, m, CH 2 ), 0.92 (3H, t, J = 7.0 Hz and 14.5 Hz, CH 3 ).
단계 2 : Step 2: NN -페닐--Phenyl- NN -- (프로-2인-1-일)펜탄아마이드(Pro-2yn-1-yl) pentaneamide (( NN -phenyl--phenyl- NN -(prop-2--(prop-2- ynyn -1-yl)pentanamide)의 제조-1-yl) pentanamide)
상기 단계 1에서 수득한 N-페닐펜탄아마이드(19.10 g, 107.40 mmol)를 N,N-다이메틸포름아마이드(DMF)(100 ml)에 녹이고 반응계를 질소 치환한 후 수소화 나트륨(sodium hydride)(5.20 g, 214.80 mmol)을 영하에서 가한 후, 2시간 동안 교반하였다. 상기 혼합물에 브로민화 프로파르길(propagyl bromide)(18.10 ml, 214.80 mmol) 을 가하고, 영하에서 2시간 동안 교반하였다. 반응액에 영하에서 물을 가한 후, 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=9:1)로 정제하여 N-페닐-N-(프로-2인-1-일)펜탄아마이드(N-phenyl-N-(prop-2-yn-1-yl)pentanamide)(19.20 g, 83% 수율). N -phenylpentaneamide (19.10 g, 107.40 mmol) obtained in step 1 was dissolved in N, N -dimethylformamide (DMF) (100 ml), the reaction system was nitrogen-substituted, and sodium hydride (5.20). g, 214.80 mmol) was added below freezing and then stirred for 2 hours. Propagyl bromide (18.10 ml, 214.80 mmol) was added to the mixture, followed by stirring for 2 hours at -40 ° C. Water was added to the reaction solution below freezing, diluted with ethyl acetate, and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. Silica gel, the concentrate was purified by column chromatography (Hex: EA = 9: 1 ) to give the N - phenyl - N - (Pro -2 a-1-yl) pentane amide (N -phenyl- N - (prop- 2-yn -1-yl) pentanamide) (19.20 g, 83% yield).
1H NMR (CDCl3, 500MHz) δ 7.42 (2H, m, aromatic), 7.37 (1H, d, J = 7.0 Hz, aromatic), 7.26 (2H, m, aromatic), 4.47 (2H, d, J = 2.0 Hz, CH2), 2.03 (2H, t, J = 7.0 Hz and 15.5 Hz, CH2), 1.53 (2H, m, CH2), 1.20 (2H, m, CH2), 0.77 (3H, t, J = 7.5 Hz and 15 Hz, CH3). 1 H NMR (CDCl 3 , 500 MHz) δ 7.42 (2H, m, aromatic), 7.37 (1H, d, J = 7.0 Hz, aromatic), 7.26 (2H, m, aromatic), 4.47 (2H, d, J = 2.0 Hz, CH 2 ), 2.03 (2H, t, J = 7.0 Hz and 15.5 Hz, CH 2 ), 1.53 (2H, m, CH 2 ), 1.20 (2H, m, CH 2 ), 0.77 (3H, t , J = 7.5 Hz and 15 Hz, CH 3 ).
단계 3 : Step 3: terttert -부틸 4-아이도벤조에이트(-Butyl 4-idobenzoate ( terttert -butyl 4-iodobenzoate)의 제조-butyl 4-iodobenzoate)
4-요오도벤조산(4-iodobenzoic acid)(1.00 g, 4.00 mmol)에 염화 티오닐(thionyl chloride)(2.30 ml, 32.30 mmol)과 N,N-다이메틸포름아마이드(DMF)(15 ml)을 N,N-다이메틸포름아마이드(DMF)(0.02 ml, 0.20mmol)를 가한 후, 반응계를 질소 치환한 후, 75℃로 가열 환류하여 1시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 테트라하이드로퓨란(tetrahydrofuran)(5ml)에 녹인 후 테트라하이드로퓨란에 녹인 포타슘 tert-부톡사이드 1몰 용액(potassium tert-butoxide 1M solution in THF)(4.5 ml)을 영하에서 천천히 가하고 같은 온도에서 반시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=9:1)로 정제하여 2-(트리메틸시릴)에틸 4-아이오도벤조에이트(2-(trimethylsilyl)ethyl 4-iodobenzoate)를 수득하였다(14.00 g, 99.9% 수율).In 4-iodobenzoic acid (1.00 g, 4.00 mmol), thionyl chloride (2.30 ml, 32.30 mmol) and N, N -dimethylformamide (DMF) (15 ml) were added. After adding N, N -dimethylformamide (DMF) (0.02 ml, 0.20 mmol), the reaction system was nitrogen-substituted and heated to reflux at 75 ° C. for 1 hour. The residue was The reaction mixture was concentrated under reduced pressure and the resulting tetrahydrofuran (tetrahydrofuran) (5ml), potassium tert- butoxide was dissolved in tetrahydrofuran, was dissolved in 1 molar solution of (1M potassium tert -butoxide solution in THF) (4.5 ml) of Slowly added below freezing and stirred at the same temperature for half hour. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 9: 1) to give 2- (trimethylsilyl) ethyl 4-iodobenzoate (14.00 g, 99.9% yield).
1H NMR (CDCl3, 400 MHz) δ 7.77 (2H, d, J = 7.5 Hz, aromatic), 7.69 (2H, d, J = 8.0 Hz, aromatic), 1.59 (9H, s, (CH3)3). 1 H NMR (CDCl 3 , 400 MHz) δ 7.77 (2H, d, J = 7.5 Hz, aromatic), 7.69 (2H, d, J = 8.0 Hz, aromatic), 1.59 (9H, s, (CH 3 ) 3 ).
단계 4 : Step 4: terttert -부틸 4--Butyl 4- (3-((3- ( NN -페닐펜탄아미도)프로프Phenylpentane amido) prop -1-인-1-일)-1-yn-1-yl) 벤조에이Benzoa 트(( terttert -butyl 4-(3-(-butyl 4- (3- ( NN -phenylpentanamido)prop-1-yn-1-yl)benzoate)의 제조-phenylpentanamido) prop-1-yn-1-yl) benzoate)
상기 단계 3에서 수득한 tert-부틸 4-아이도벤조에이트(4.90 g, 16.00 mmol)를 테트라하이드로퓨란(tetrahydrofuran)(30ml)에 녹인 용액에 상기 단계 2에서 수득한 N-페닐-N-(프로-2인-1-일)펜탄아마이드(2.30 g, 10.70 mmol)을 가하고 반응계를 질소 치환한 후 상온에서 5분간 교반했다. 상기 혼합물에 트리에틸아민(24 ml), 비스(트리페닐포스핀)팔라듐(II) 다이클로라이드(75.00 mg, 0.10 mmol)와 요오드화 구리(I)(41.00 mg, 0.21 mmol)를 가하고 상온에서 16시간 동안 교반하였다. 감압 하에 농축하고 얻어진 잔사를 다이클로로메탄으로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=4:1)로 정제하여 tert-부틸 4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤조에이트(tert-butyl 4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzoate) (3.30 g, 78.3% 수율). N -phenyl- N- (pro) obtained in step 2 in a solution of tert -butyl 4-idbenzoate (4.90 g, 16.00 mmol) obtained in step 3 in tetrahydrofuran (30 ml) -2yn-1-yl) pentaneamide (2.30 g, 10.70 mmol) was added thereto, the reaction system was replaced with nitrogen, and stirred at room temperature for 5 minutes. Triethylamine (24 ml), bis (triphenylphosphine) palladium (II) dichloride (75.00 mg, 0.10 mmol) and copper iodide (I) (41.00 mg, 0.21 mmol) were added to the mixture for 16 hours at room temperature. Was stirred. The residue obtained by concentration under reduced pressure was diluted with dichloromethane and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 4: 1) to give tert -butyl 4- (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzoate (tert -butyl 4- (3- ( N- phenylpentanamido) prop-1-yn-1-yl) benzoate) (3.30 g, 78.3% yield).
1H-NMR (500MHz, CDCl3): δ 7.88 (2H, d, J = 8.0 Hz, aromatic), 7.45 (2H, m, aromatic), 7.39 (1H, d, J = 7.0 Hz, aromatic), 7.35 (2H, d, J = 8.0 Hz, aromatic), 7.30 (2H, d, J = 5.0 Hz, aromatic), 4.72 (2H, s, CH2), 2.07 (2H, t, J = 7.5 Hz and 15 Hz, CH2), 1.56 (2H, m, CH2), 1.51 (9H, s, (CH3)3), 1.22 (2H, m, CH2), 0.80 (3H, t, 7.5 Hz and 15 Hz7.5 Hz and 15 Hz, CH3). 1 H-NMR (500 MHz, CDCl 3 ): δ 7.88 (2H, d, J = 8.0 Hz, aromatic), 7.45 (2H, m, aromatic), 7.39 (1H, d, J = 7.0 Hz, aromatic), 7.35 (2H, d, J = 8.0 Hz, aromatic), 7.30 (2H, d, J = 5.0 Hz, aromatic), 4.72 (2H, s, CH 2 ), 2.07 (2H, t, J = 7.5 Hz and 15 Hz , CH 2 ), 1.56 (2H, m, CH 2 ), 1.51 (9H, s, (CH 3 ) 3 ), 1.22 (2H, m, CH 2 ), 0.80 (3H, t, 7.5 Hz and 15 Hz 7. 5 Hz and 15 Hz, CH 3 ).
실시예Example 38. 4- 38. 4- (3-((3- ( NN -페닐펜탄아미도)프로-1-핀-1-일)벤조익산-Phenylpentaneamido) prop-1-pin-1-yl) benzoic acid (4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzoic acid)의 제조(LMT-1013)Preparation of (4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzoic acid) (LMT-1013)
상기 실시예 37에서 수득한 tert-부틸 4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤조에이트(tert-butyl 4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzoate)(2.00 g, 5.10 mmol)를 아세토나이트릴(48 ml)에 녹인 후, 영하에서 5분 동안 교반하였다. 상기 용액에 트리프루오로아세틱산(trifluoroacetic acid)(12ml)를 천천히 가하고, 상온에서 48시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산 마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기농축액을 실리카겔 컬럼 크로마토그래피(HEX:EA=2:1)로 정제하여 4-(3-(N-페닐펜탄아미도)프로-1-핀-1-일)벤조익산(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzoic acid)을 수득하였다(1.6 g, 95% 수율).Tert-butyl 4- (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzoate obtained in Example 37 (tert-butyl 4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzoate) (2.00 g, 5.10 mmol) was dissolved in acetonitrile (48 ml) and stirred for 5 minutes at zero. Trifluoroacetic acid (12 ml) was slowly added to the solution, followed by stirring at room temperature for 48 hours. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (HEX: EA = 2: 1) to give 4- (3- (N-phenylpentaneamido) prop-1-pin-1-yl) benzoic acid (4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzoic acid) was obtained (1.6 g, 95% yield).
1H-NMR (500 MHz, MeOD): δ 7.95 (2H, d, J = 8.5 Hz, aromatic), 7.51 (2H, m, aromatic), 7.45 (1H, m, aromatic), 7.44 (2H, d, J = 8.5 Hz, aromatic), 7.39 (2H, d, J = 8.0 Hz, aromatic), 4.73 (2H, s, CH2), 2.11 (2H, t, CH3), 1.21 (2H, m, CH2), 0.83 (3H, t, CH3). 1 H-NMR (500 MHz, MeOD): δ 7.95 (2H, d, J = 8.5 Hz, aromatic), 7.51 (2H, m, aromatic), 7.45 (1H, m, aromatic), 7.44 (2H, d, J = 8.5 Hz, aromatic), 7.39 (2H, d, J = 8.0 Hz, aromatic), 4.73 (2H, s, CH2), 2.11 (2H, t, CH3), 1.21 (2H, m, CH2), 0.83 (3H, t, CH 3).
실시예Example 39. N-에틸-4- 39.N-ethyl-4- (3-(N-페닐펜탄아미도)프로프(3- (N-phenylpentaneamido) prop -1-인-1-일)-1-yn-1-yl) 벤즈아마이Benz Amai 드(N-ethyl-4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamide)의 제조 (Preparation of de (N-ethyl-4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamide) LMTLMT -1017)-1017)
상기 실시예 38에서 수득한 4-(3-(N-페닐펜탄아미도)프로-1-핀-1-일)벤조익산(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzoic acid)(80.00 mg, 0.24 mmol)를 N,N-다이메틸포름아마이드(DMF)(0.70 ml)에 녹이고 트리에틸아민(0.70 ml)에서 1시간 동안 교반시킨 에틸아민하이드로클로라이드(ethylamine ydrochloride)(29.20 mg, 0.36mmol)와 1-하이드록시벤보트리아졸 하이드레이트(1-hydroxybenzotriazole hydrate)(48.4 mg, 0.36 mmol)와 1-에틸-3-(3-다이베틸아미노프로필)칼보다이이마이드 하이드로클로라이드(1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride)(55.5 mg, 0.36 mmol)를 상기 용액에 가하고, 상온애서 16시간 동안 교반하였다. 반응액을 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산 마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=1:1)로 정제하여 N-에틸-4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아마이드(N-ethyl-4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamide)를 수득하였다(56.8 mg, 66% 수율).4- (3- (N-phenylpentaneamido) prop-1-pin-1-yl) benzoic acid obtained in Example 38 (4- (3- (N-phenylpentanamido) prop-1-yn-1 -yl) benzoic acid) (80.00 mg, 0.24 mmol) was dissolved in N, N-dimethylformamide (DMF) (0.70 ml) and stirred in triethylamine (0.70 ml) for 1 hour. ydrochloride) (29.20 mg, 0.36 mmol) and 1-hydroxybenzotriazole hydrate (48.4 mg, 0.36 mmol) and 1-ethyl-3- (3-dibutylaminopropyl) Amide hydrochloride (1-Ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride) (55.5 mg, 0.36 mmol) was added to the solution and stirred for 16 hours at room temperature. The reaction solution was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 1: 1) to obtain N-ethyl-4- (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzamide ( N-ethyl-4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamide) was obtained (56.8 mg, 66% yield).
1H NMR (CDCl3, 500 MHz) δ 7.69 (2H, d, J = 8.5 Hz, aromatic), 7.39 (7H, m, aromatic), 4.72 (2H, s, CH2), 3.49 (2H, t, J = 6.0 Hz and 13.0 Hz, CH2), 2.08 (2H, t, J =7.5 Hz and 15.0 Hz, CH2), 1.57 (2H, m, CH2), 1.23 (2H, m, CH2), 0.81 (3H, t, J = 7.5 Hz and 14.5 Hz, CH3). 1 H NMR (CDCl 3 , 500 MHz) δ 7.69 (2H, d, J = 8.5 Hz, aromatic), 7.39 (7H, m, aromatic), 4.72 (2H, s, CH 2), 3.49 (2H, t, J = 6.0 Hz and 13.0 Hz, CH2), 2.08 (2H, t, J = 7.5 Hz and 15.0 Hz, CH2), 1.57 (2H, m, CH2), 1.23 (2H, m, CH2), 0.81 (3H, t , J = 7.5 Hz and 14.5 Hz, CH3).
실시예Example 40. N-(2-(다이에틸아미노)에틸)-4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아마이드(N-(2-(dimethylamino)ethyl)-4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamide)의 제조(LMT-1016) 40.N- (2- (diethylamino) ethyl) -4- (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzamide (N- (2- (dimethylamino) Preparation of ethyl) -4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamide) (LMT-1016)
상기 실시예 38에서 수득한 4-(3-(N-페닐펜탄아미도)프로-1-핀-1-일)벤조익산(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzoic acid)(80.00 mg, 0.24 mmol)에 N,N-다이메틸에테인-1,2-다이아민(N,N-dimethylethane-1,2-diamine)(0.04 ml, 0.36 mmol)을 이용하여 실시예 39와 동일한 방법으로 N-(2-(다이에틸아미노)에틸)-4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아마이드(N-(2-(dimethylamino)ethyl)-4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamide)를 수득하였다(71.72 mg, 74% 수율).4- (3- (N-phenylpentaneamido) prop-1-pin-1-yl) benzoic acid obtained in Example 38 (4- (3- (N-phenylpentanamido) prop-1-yn-1 -yl) benzoic acid) (80.00 mg, 0.24 mmol) using N, N-dimethylethane-1,2-diamine (0.04 ml, 0.36 mmol) N- (2- (diethylamino) ethyl) -4- (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzamide (N- (2- (dimethylamino) ethyl) -4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamide) was obtained (71.72 mg, 74% yield).
1H NMR (CDCl3, 500 MHz) δ 7.71 (2H, d, J = 8.0 Hz, aromatic), 7.38 (7H, m, aromatic), 4.70 (2H, s, CH2), 3.48 (2H, m, CH2), 2.50 (2H, t, J = 6.0 Hz and 11.5 Hz, CH2), 2.24 (6H, s, (CH3)2), 2.05 (2H, t, J = 7.5 Hz and 15.0 Hz, CH2), 1.54 (2H, m, CH2), 1.20 (2H, m, CH2), 0.78 (3H, t, J = 7.0 Hz and 14.0 Hz, CH3). 1 H NMR (CDCl 3 , 500 MHz) δ 7.71 (2H, d, J = 8.0 Hz, aromatic), 7.38 (7H, m, aromatic), 4.70 (2H, s, CH 2), 3.48 (2H, m, CH 2 ), 2.50 (2H, t, J = 6.0 Hz and 11.5 Hz, CH2), 2.24 (6H, s, (CH3) 2), 2.05 (2H, t, J = 7.5 Hz and 15.0 Hz, CH2), 1.54 ( 2H, m, CH 2), 1.20 (2H, m, CH 2), 0.78 (3H, t, J = 7.0 Hz and 14.0 Hz, CH 3).
실시예Example 41. 에틸 2-(4-(3-(N- 41. Ethyl 2- (4- (3- (N- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1-인-1-일)-1-yn-1-yl) 벤즈아미도Benzamido )아세테이트(ethyl 2-(4-(3-(N-Acetate (ethyl 2- (4- (3- (N-) phenylpentanamidophenylpentanamido )prop-1-prop-1- ynyn -1-yl)benzamido)acetate)의 제조 (LMT-1014)-1-yl) benzamido) acetate) (LMT-1014)
상기 실시예 38에서 수득한 4-(3-(N-페닐펜탄아미도)프로-1-핀-1-일)벤조익산(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzoic acid)(150.00 mg, 0.45 mmol)에 글리신에틸에스터 하이드로클로라이드(glycineethylester hydrochloride)(93.70 mg, 0.67 mmol)을 이용하여 실시예 39와 동일한 방법으로 에틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)아세테이트(ethyl 2-(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamido)acetate)를 수득하였다(102.18 mg, 54% 수율).4- (3- (N-phenylpentaneamido) prop-1-pin-1-yl) benzoic acid obtained in Example 38 (4- (3- (N-phenylpentanamido) prop-1-yn-1 -yl) benzoic acid) (150.00 mg, 0.45 mmol) using glycineethylester hydrochloride (93.70 mg, 0.67 mmol) in the same manner as in Example 39 to obtain ethyl 2- (4- (3- ( N-phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetate (ethyl 2- (4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamido) acetate) was obtained (102.18 mg, 54% yield).
1H NMR (CDCl3, 500 MHz) δ 7.74 (2H, d, J = 8.0 Hz, aromatic), 7.37 (7H, m, aromatic), 4.22 (4H, m, (CH2)2), 2.07 (2H, t, J = 7.5 Hz and 15.0 Hz, CH2), 1.56 (2H, m, CH2), 1.30 (5H, m, CH3, CH2), 1.22 (2H, m, CH2), 0.80 (3H, t, J = 7.5 Hz and 14.5 Hz, CH3). 1 H NMR (CDCl 3 , 500 MHz) δ 7.74 (2H, d, J = 8.0 Hz, aromatic), 7.37 (7H, m, aromatic), 4.22 (4H, m, (CH 2) 2), 2.07 (2H, t, J = 7.5 Hz and 15.0 Hz, CH2), 1.56 (2H, m, CH2), 1.30 (5H, m, CH3, CH2), 1.22 (2H, m, CH2), 0.80 (3H, t, J = 7.5 Hz and 14.5 Hz, CH 3).
실시예Example 42. 2- 42.2- (4-(3-(N-페닐펜탄아미도)프로프(4- (3- (N-phenylpentaneamido) prop -1-인-1-일)-1-yn-1-yl) 벤즈아미도Benzamido )) 아세틱Acetic 산(2-(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamido)acetic acid)의 제조(LMT-1015)Preparation of acid (2- (4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamido) acetic acid) (LMT-1015)
상기 실시예 41에서 수득한 에틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)아세테이트(ethyl 2-(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamido)acetate)(46.20 mg, 0.10 mmol)와 나트륨 하이드록사이드 2몰 수용액(0.07 ml, 0.14 mmol)을 메탄올(0.1ml)에 녹인 후 상온에서 30분간 교반하였다. 염산을 이용해서 반응액의 산성도를 높인 후 에틸아세테이트로 희석하고, 물과 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산 마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 헥센과 에틸아세테이트를 이용하여 재결정한 후 2-(4(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)아세틱 산(2-(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamido)acetic acid)를 수득하였다(20.80 mg, 53% 수율).Ethyl 2- (4- (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetate obtained in Example 41 (ethyl 2- (4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamido) acetate) (46.20 mg, 0.10 mmol) and a 2-mol aqueous solution of sodium hydroxide (0.07 ml, 0.14 mmol) were dissolved in methanol (0.1 ml). Stirred at room temperature for 30 minutes. The acidity of the reaction solution was increased using hydrochloric acid, diluted with ethyl acetate, and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4), filtered and concentrated under reduced pressure. The concentrate was recrystallized with hexene and ethyl acetate and then 2- (4 (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetic acid (2- ( 4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamido) acetic acid) was obtained (20.80 mg, 53% yield).
1H NMR (CDCl3 , 500 MHz) δ 7.71 (2H, d, J = 8.5 Hz, aromatic), 7.40 (7H, m, aromatic), 4.71 (2H, s, CH2), 4.23 (2H, d, J = 5.0 Hz, CH2), 2.10 (2H, t, J = 7.5 Hz and 15.5 Hz, CH2), 1.56 (2H, m, CH2), 1.22 (2H, m, CH2), 0.80 (3H, t, J = 7.0 Hz and 14.5 Hz, CH3). 1 H NMR (CDCl 3 , 500 MHz) δ 7.71 (2H, d, J = 8.5 Hz, aromatic), 7.40 (7H, m, aromatic), 4.71 (2H, s, CH 2), 4.23 (2H, d, J = 5.0 Hz, CH2), 2.10 (2H, t, J = 7.5 Hz and 15.5 Hz, CH2), 1.56 (2H, m, CH2), 1.22 (2H, m, CH2), 0.80 (3H, t, J = 7.0 Hz and 14.5 Hz, CH 3).
실시예Example 43.  43. 메틸methyl 2-(4-(3-(N- 2- (4- (3- (N- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1-인-1-일)-1-yn-1-yl) 벤즈아미도Benzamido )프로파노에이트(methyl 2-(4-(3-(N-Propanoate (methyl 2- (4- (3- (N-) phenylpentanamidophenylpentanamido )prop-1-prop-1- ynyn -1-yl)benzamido)propanoate)의 제조 (LMT-1018)-1-yl) benzamido) propanoate) (LMT-1018)
상기 실시예 38에서 수득한 4-(3-(N-페닐펜탄아미도)프로-1-핀-1-일)벤조익산(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzoic acid)(100.00 mg, 0.30 mmol)에 L-알라닌메틸에스터 하이드로클로라이드(L-alaninemethylester hydrochloride)(83.70 mg, 0.60 mmol)을 이용하여 실시예 39와 동일한 방법으로 메틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)프로파노에이트(methyl 2-(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamido)propanoate)를 수득하였다(110.00 mg, 42% 수율).4- (3- (N-phenylpentaneamido) prop-1-pin-1-yl) benzoic acid obtained in Example 38 (4- (3- (N-phenylpentanamido) prop-1-yn-1 L-alaninemethylester hydrochloride (83.70 mg, 0.60 mmol) in -yl) benzoic acid) (100.00 mg, 0.30 mmol) in the same manner as in Example 39. (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzamido) propanoate (methyl 2- (4- (3- (N-phenylpentanamido) prop-1-yn- 1-yl) benzamido) propanoate) was obtained (110.00 mg, 42% yield).
1H NMR (CDCl3 , 500 MHz) δ 7.72 (2H, d, J = 8.5 Hz, aromatic), 7.37 (7H, m, aromatic), 4.76 (1H, t, J = 7.5 Hz and 14.5 Hz CH), 4.71 (2H, s, CH2), 3.76 (3H, s, CH3), 2.06 (2H, t, J = 7.5 Hz and 15.5 Hz, CH2), 1.53 (2H, m, CH2), 1.50 (3H, d, J = 7.5 Hz, CH3), 1.20 (2H, m, CH2), 0.79 (3H, t, J = 7.5 Hz and 15.0 Hz, CH3). 1 H NMR (CDCl 3 , 500 MHz) δ 7.72 (2H, d, J = 8.5 Hz, aromatic), 7.37 (7H, m, aromatic), 4.76 (1H, t, J = 7.5 Hz and 14.5 Hz CH), 4.71 (2H, s, CH2), 3.76 (3H, s, CH3), 2.06 (2H, t, J = 7.5 Hz and 15.5 Hz, CH2), 1.53 (2H, m, CH2), 1.50 (3H, d, J = 7.5 Hz, CH 3), 1.20 (2H, m, CH 2), 0.79 (3H, t, J = 7.5 Hz and 15.0 Hz, CH 3).
실시예Example 44. 2-(4-(3-(N- 44. 2- (4- (3- (N- 페닐펜탄아미도Phenylpentaneamido )) 프로프Prof -1-인-1-일)-1-yn-1-yl) 벤즈아미도Benzamido )) 프로피오닉산Propionic acid (2-(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamido)propanoic acid)의 제조(LMT-1019)Preparation of (2- (4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamido) propanoic acid) (LMT-1019)
상기 실시예 43에서 수득한 메틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)프로파노에이트(methyl 2-(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamido)propanoate)(86.50 mg, 0.20 mmol)를 이용하여 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)프로피오닉 산(2-(4-(3-(N-phenylpentanamido)prop-1-yn-1-yl)benzamido)propanoic acid)를 수득하였다(45.00 mg, 55% 수율).Methyl 2- (4- (3- (N-phenylpentaneamido) prop-1-yn-1-yl) benzamido) propanoate obtained in Example 43 (methyl 2- (4- ( 2- (4- (3- (N-phenylpentaneamido) prop- with 3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamido) propanoate) (86.50 mg, 0.20 mmol) 1-yn-1-yl) benzamido) propionic acid (2- (4- (3- (N-phenylpentanamido) prop-1-yn-1-yl) benzamido) propanoic acid) was obtained (45.00 mg) , 55% yield).
1H NMR (CDCl3, 500 MHz) δ 7.71 (2H, d, J = 8.0 Hz, aromatic), 7.40 (7H, m, aromatic), 4.76 (1H, t, J = 7.5 Hz and 14.5 Hz, CH), 4.72 (2H, s, CH2), 2.10 (2H, t, J = 7.5 Hz and 15.5 Hz, CH2), 1.57 (5H, m, CH3, CH2), 1.22 (2H, m, CH2), 0.80 (3H, t, J = 7.5 Hz and 15.0 Hz, CH3). 1 H NMR (CDCl 3 , 500 MHz) δ 7.71 (2H, d, J = 8.0 Hz, aromatic), 7.40 (7H, m, aromatic), 4.76 (1H, t, J = 7.5 Hz and 14.5 Hz, CH) , 4.72 (2H, s, CH2), 2.10 (2H, t, J = 7.5 Hz and 15.5 Hz, CH2), 1.57 (5H, m, CH3, CH2), 1.22 (2H, m, CH2), 0.80 (3H , t, J = 7.5 Hz and 15.0 Hz, CH3).
실시예Example 45. 2-(4-(3-(N-(3- 45. 2- (4- (3- (N- (3- 플루오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )) 페녹시Phenoxy )아세트산(2-(4-(3-(N-(3-fluorophenyl)pentanamido)prop-1-ynyl)phenoxy)acetic acid)의 제조(LMT-1009)) Preparation of acetic acid (2- (4- (3- (N- (3-fluorophenyl) pentanamido) prop-1-ynyl) phenoxy) acetic acid) (LMT-1009)
단계 1 : N-(3-Step 1: N- (3- 플루오로페닐Fluorophenyl )) 펜탄아마이드Pentaneamide (N-(3- (N- (3- fluorophenylfluorophenyl )) pentanamidepentanamide )의 제조Manufacturing
3-플루오로아닐린(3-fluoroaniline)(200.00 mg, 1.79 mmol)를 이용하여 실시예 37의 단계 1과 동일한 방법으로 N-(3-플루오로페닐)펜탄아마이드 (N-(3-fluorophenyl)pentanamide)를 수득하였다(349.00 mg, 99% 수율).N- (3-fluorophenyl) pentanamide (N- (3-fluorophenyl) pentanamide in the same manner as in Step 1 of Example 37 using 3-fluoroaniline (200.00 mg, 1.79 mmol) ) Was obtained (349.00 mg, 99% yield).
1H NMR (CDCl3, 500 MHz) δ 8.10 (1H, s), 7.51 (1H, d, J = 11.0 Hz), 7.20 (2H, m), 6.79 (1H, m), 2.36 (2H, t), 1.68 (2H, m), 1.36 (2H, m), 0.91 (3H, t) 1 H NMR (CDCl 3 , 500 MHz) δ 8.10 (1H, s), 7.51 (1H, d, J = 11.0 Hz), 7.20 (2H, m), 6.79 (1H, m), 2.36 (2H, t) , 1.68 (2H, m), 1.36 (2H, m), 0.91 (3H, t)
단계 2 : N-(3-Step 2: N- (3- 플루오로페틸FluoroPetyl )-N-() -N- ( 프로프Prof -2--2- 이닐Inil )) 펜탄아마이드Pentaneamide (N-(3-fluorophenyl)-N-(prop-2-ynyl)pentanamide)의 제조 Preparation of (N- (3-fluorophenyl) -N- (prop-2-ynyl) pentanamide)
상기 단계 1에서 수득한 N-(3-플루오로페닐)펜탄아마이드 (400.00 mg, 2.05 mmol)와 포타시움하이드록사이드(Potassium hydroxide)(230.61 mg, 4.11 mmol), 테트라부틸암모니움아이오다이드(tetrabutyl ammonium Iodide)(37.87 mg, 0.20 mmol)을 테트라하이드로퓨란(Tetrahydrofuran)(20.00 ml)에 녹인 후 20분 동안 교반하였다. 같은 온도에서 상기 혼합물에 프로파질브로마이드(propargyl bromide)(0.19 ml, 2.30 mmol)을 가한 뒤 20시간 동안 교반하였다. 반응액을 감압 하에 농축하고 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 실리카겔 컬럼 크로마토그래피(Hex:EA=10:1)로 정제하여 N-(3-플루오로페틸)-N-(프로프-2-이닐)펜탄아마이드 (N-(3-fluorophenyl)-N-(prop-2-ynyl)pentanamide)를 수득하였다(450.00 mg, 94% 수율).N- (3-fluorophenyl) pentaneamide (400.00 mg, 2.05 mmol) and Potassium hydroxide (230.61 mg, 4.11 mmol) obtained in step 1, tetrabutylammonium iodide (tetrabutyl) ammonium Iodide) (37.87 mg, 0.20 mmol) was dissolved in Tetrahydrofuran (20.00 ml) and stirred for 20 minutes. At the same temperature, propargyl bromide (0.19 ml, 2.30 mmol) was added to the mixture, followed by stirring for 20 hours. The reaction solution was concentrated under reduced pressure and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4), filtered and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (Hex: EA = 10: 1) to obtain N- (3-fluoropetyl) -N- (prop-2-ynyl) pentaneamide (N- (3-fluorophenyl)- N- (prop-2-ynyl) pentanamide) was obtained (450.00 mg, 94% yield).
1H NMR (CDCl3, 500 MHz) δ 7.42 (1H, m), 7.11 (2H, m), 7.20 (1H, d, J = 9.0 Hz ), 4.46 (2H, s), 2.07 (2H, t), 1.56 (2H, m), 1.22 (2H, m), 0.82 (3H, t)1 H NMR (CDCl 3, 500 MHz) δ 7.42 (1H, m), 7.11 (2H, m), 7.20 (1H, d, J = 9.0 Hz), 4.46 (2H, s), 2.07 (2H, t), 1.56 (2H, m), 1.22 (2H, m), 0.82 (3H, t)
단계 3 : N-(3-Step 3: N- (3- 플루오로페닐Fluorophenyl )-N-(3-(4-) -N- (3- (4- 하이드록시페닐Hydroxyphenyl )) 프로프Prof -2--2- 이닐Inil )) 펜탄아Pentanah 마이드 (N-(3-Amide (N- (3- fluorophenylfluorophenyl )-N-(3-(4-) -N- (3- (4- hydroxyphenylhydroxyphenyl )prop-2-prop-2- ynylynyl )) pentanamidepentanamide )의 제조Manufacturing
상기 단계 2에서 수득한 N-(3-플루오로페틸)-N-(프로프-2-이닐)펜탄아마이드 (270.00 mg, 1.16 mmol)와 4-아이오도페놀(4-Iodophenol)(127.60 mg, 0.58 mmol)을 이용하여 N-(3-플루오로페닐)-N-(3-(4-하이드록시페닐)프로프-2-이닐)펜탄아마이드 (N-(3-fluorophenyl)-N-(3-(4-hydroxyphenyl)prop-2-ynyl)pentanamide)를 수득하였다(128.00 mg, 67.8% 수율).N- (3-fluorofetyl) -N- (prop-2-ynyl) pentaneamide (270.00 mg, 1.16 mmol) and 4-iodophenol (127.60 mg, obtained in step 2) 0.58 mmol) with N- (3-fluorophenyl) -N- (3- (4-hydroxyphenyl) prop-2-ynyl) pentaneamide (N- (3-fluorophenyl) -N- (3 -(4-hydroxyphenyl) prop-2-ynyl) pentanamide) was obtained (128.00 mg, 67.8% yield).
1H NMR (CDCl3, 500 MHz) δ 7.51 (1H, m), 7.21 (5H, m), 6.71 (2H, d, J = 9.0 Hz), 7.65 (2H, s), 2.13 (2H, t), 1.54 (2H, m), 1.26 (2H, m), 0.82 (3H, t) 1 H NMR (CDCl 3 , 500 MHz) δ 7.51 (1H, m), 7.21 (5H, m), 6.71 (2H, d, J = 9.0 Hz), 7.65 (2H, s), 2.13 (2H, t) , 1.54 (2H, m), 1.26 (2H, m), 0.82 (3H, t)
단계 4 : 에틸 2-(4-(3-(N-(3-Step 4: ethyl 2- (4- (3- (N- (3- 플루오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )) 페녹시Phenoxy )아세테이트 (ethyl 2-(4-(3-(N-(3-Acetate (ethyl 2- (4- (3- (N- (3- fluorophenylfluorophenyl )) pentanamidopentanamido )prop-1-ynyl)phenoxy)acetate)의 제조) prop-1-ynyl) phenoxy) acetate)
상기 단계 3에서 수득한 N-(3-플루오로페닐)-N-(3-(4-하이드록시페닐)프로프-2-이닐)펜탄아마이드 (120.00mg, 0.37 mmol)와 포타시움카보네이트(Potassium carbonate)(153.41 mg, 1.11 mmol)를 에틸 2-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세테이트 (ethyl 2-(4-(3-(N-(3-fluorophenyl)pentanamido)prop-1-ynyl)phenoxy)acetate)를 수득하였다(113.00 mg, 74% 수율).N- (3-fluorophenyl) -N- (3- (4-hydroxyphenyl) prop-2-ynyl) pentaneamide (120.00 mg, 0.37 mmol) and Potassium carbonate obtained in step 3 above ) (153.41 mg, 1.11 mmol) in ethyl 2- (4- (3- (N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy) acetate (ethyl 2- (4- (3- (N- (3-fluorophenyl) pentanamido) prop-1-ynyl) phenoxy) acetate) was obtained (113.00 mg, 74% yield).
1H NMR (CDCl3, 500 MHz) δ 7.42 (1H, m), 7.28 (2H, d, J = 9.0 Hz), 7.09 (3H, m), 6.81 (2H, d, J = 9.0 Hz), 4.68 (2H, s), 4.61 (2H, s), 4.27 (2H, m), 2.05 (2H, t), 1.58 (2H, m), 1.26 (5H, m), 0.82 (3H, t) 1 H NMR (CDCl 3 , 500 MHz) δ 7.42 (1H, m), 7.28 (2H, d, J = 9.0 Hz), 7.09 (3H, m), 6.81 (2H, d, J = 9.0 Hz), 4.68 (2H, s), 4.61 (2H, s), 4.27 (2H, m), 2.05 (2H, t), 1.58 (2H, m), 1.26 (5H, m), 0.82 (3H, t)
단계 step 5 : 25: 2 -(4-(3-(N-(3--(4- (3- (N- (3- 플루오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )) 페녹시Phenoxy )) 아세틱산Acetic acid (2-(4-(3-(N-(3-fluorophenyl)pentanamido)prop-1-ynyl)phenoxy)acetic acid)의 제조Preparation of (2- (4- (3- (N- (3-fluorophenyl) pentanamido) prop-1-ynyl) phenoxy) acetic acid)
상기 단계 4에서 수득한 에틸 2-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세테이트 (100.00 mg, 0.24 mmol)을 에탄올(Ethanol)(9.00 ml)에 녹인 후, 5분 동안 교반하였다. 상기 혼합물에 소듐하이드록사이드 2M (Sodium hydroxide 2M)(0.30 ml)를 가하고 80oC에서 가열, 환류하여 3시간 동안 교반하였다. 반응액을 상온에서 냉각 후, 감압 하에 농축한 뒤, 얻어진 잔사를 에틸 아세테이트로 희석하고, 물 및 소금물로 세척하였다. 유기용매 층을 모아 무수의 황산마그네슘(MgSO4)으로 수분을 제거하고 여과한 뒤, 감압 하에 농축하였다. 상기 농축액을 ODS 컬럼 크로마토그래피(MeOD:H2O=2:1)로 정제하여 2-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세트산 (2-(4-(3-(N-(3-fluorophenyl)pentanamido)prop-1-ynyl)phenoxy)acetic acid)를 를 수득하였다(15.00 mg, 16% 수율)Ethyl 2- (4- (3- (N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy) acetate (100.00 mg, 0.24 mmol) obtained in step 4 was added to ethanol ( Ethanol) (9.00 ml) and stirred for 5 minutes. Sodium hydroxide 2M (0.30 ml) was added to the mixture, and the mixture was heated and refluxed at 80 ° C. for 3 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was diluted with ethyl acetate and washed with water and brine. The organic solvent layers were combined, the water was removed with anhydrous magnesium sulfate (MgSO 4), filtered and concentrated under reduced pressure. The concentrate was purified by ODS column chromatography (MeOD: H 2 O = 2: 1) to give 2- (4- (3- (N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy ) Acetic acid (2- (4- (3- (N- (3-fluorophenyl) pentanamido) prop-1-ynyl) phenoxy) acetic acid) was obtained (15.00 mg, 16% yield)
1H NMR (CDCl3, 500 MHz) δ 7.52 (1H, d, J = 7.0 Hz), 7.24 (5H, m), 6.88 (2H, d, J = 8.5 Hz), 4.68 (2H, s), 4.66 (2H, s), 2.14 (2H, t), 1.55 (2H, m), 1.24 (2H, m), 0.83 (3H, t) 1 H NMR (CDCl 3 , 500 MHz) δ 7.52 (1H, d, J = 7.0 Hz), 7.24 (5H, m), 6.88 (2H, d, J = 8.5 Hz), 4.68 (2H, s), 4.66 (2H, s), 2.14 (2H, t), 1.55 (2H, m), 1.24 (2H, m), 0.83 (3H, t)
실시예Example 46. 2-(4-(3-(N-(4- 46. 2- (4- (3- (N- (4- 플루오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )) 페녹시Phenoxy )아세트산(2-(4-(3-(N-(4-fluorophenyl)pentanamido)prop-1-ynyl)phenoxy)acetic acid)의 제조(LMT-1010)) Preparation of acetic acid (2- (4- (3- (N- (4-fluorophenyl) pentanamido) prop-1-ynyl) phenoxy) acetic acid) (LMT-1010)
단계 1 : N-(4-Step 1: N- (4- 플로오로페닐Fluorophenyl )) 펜탄아마이드Pentaneamide (N-(4- (N- (4- fluorophenylfluorophenyl )) pentanamidepentanamide )의 제조Manufacturing
4-플루오로아닐린(4-fluoroaniline)(500.00 mg, 4.49 mmol)과 발레로일 클로라이드(valeroyl chloride)(1.10 ml, 8.99 mmol)을 이용하여 상기 실시예 37의 단계 1와 동일한 방법으로 N-(4-플로오로페닐)펜탄아마이드 (N-(4-fluorophenyl)pentanamide)를 수득하였다(870.00 mg, 99 % 수율).In the same manner as in Step 1 of Example 37, 4-fluoroaniline (500.00 mg, 4.49 mmol) and valeroyl chloride (1.10 ml, 8.99 mmol) were used. 4-Fluorophenyl) pentaneamide (N- (4-fluorophenyl) pentanamide) was obtained (870.00 mg, 99% yield).
1H NMR (CDCl3, 400 MHz) δ 8.16(1H, br), 7.48-7.45(2H, m), 6.97-6.94(2H, m), 2.34-2.31(2H, t), 1.68-1.65(2H, m), 1.38-1.33(2H, m), 0.92-0.89(3H, t). 1 H NMR (CDCl 3 , 400 MHz) δ 8.16 (1H, br), 7.48-7.45 (2H, m), 6.97-6.94 (2H, m), 2.34-2.31 (2H, t), 1.68-1.65 (2H m), 1.38-1.33 (2H, m), 0.92-0.89 (3H, t).
단계 2 : N-(4-Step 2: N- (4- 플로오로페닐Fluorophenyl )-N-() -N- ( 프로프Prof -2--2- 이닐Inil )) 펜탄아마이드Pentaneamide (N-(4-fluorophenyl)-N-(prop-2-ynyl)pentanamide)의 제조 Preparation of (N- (4-fluorophenyl) -N- (prop-2-ynyl) pentanamide)
상기 단계 1에서 수득한 N-(4-플로오로페닐)펜탄아마이드 (500.00 mg, 2.56mmol)과 프로파질브로마이드(propargyl bromide) (0.24 ml, 2.81 mmol)을 이용하여 상기 실시예 45의 단계 2와 동일한 방법으로 N-(4-플로오로페닐)-N-(프로프-2-이닐)펜탄아마이드 (N-(4-fluorophenyl)-N-(prop-2-ynyl)pentanamide)를 수득하였다(347.60 mg, 57 % 수율).Example 2 of Example 45 using N- (4-fluorophenyl) pentaneamide (500.00 mg, 2.56 mmol) and propargyl bromide (0.24 ml, 2.81 mmol) obtained in step 1; In the same manner, N- (4-fluorophenyl) -N- (prop-2-ynyl) pentaneamide (N- (4-fluorophenyl) -N- (prop-2-ynyl) pentanamide) was obtained (347.60). mg, 57% yield).
1H NMR (CDCl3, 500 MHz) δ 7.27 (2H, m), 7.14 (2H, m), 4.46 (2H, s), 2.04 (2H, t), 1.55 (2H, m), 1.21 (2H, m), 0.81 (3H, t)1 H NMR (CDCl 3, 500 MHz) δ 7.27 (2H, m), 7.14 (2H, m), 4.46 (2H, s), 2.04 (2H, t), 1.55 (2H, m), 1.21 (2H, m) , 0.81 (3H, t)
단계 3 : N-(4-Step 3: N- (4- 플로오로페닐Fluorophenyl )-N-(3-(4-) -N- (3- (4- 하이드록시페닐Hydroxyphenyl )) 프로프Prof -2--2- 이닐Inil )) 펜탄아Pentanah 마이드 (N-(4-Amide (N- (4- fluorophenylfluorophenyl )-N-(3-(4-) -N- (3- (4- hydroxyphenylhydroxyphenyl )prop-2-prop-2- ynylynyl )) pentanamidepentanamide )의 제조Manufacturing
상기 단계 3에서 수득한 N-(4-플로오로페닐)-N-(프로프-2-이닐)펜탄아마이드 (300.00 mg, 1.29 mmol)과 4-아이오도페놀(4-Iodophenol)(140.80 mg, 0.64 mmol)을 이용하여 N-(4-플로오로페닐)-N-(3-(4-하이드록시페닐)프로프-2-이닐)펜탄아마이드 (N-(4-fluorophenyl)-N-(3-(4-hydroxyphenyl)prop-2-ynyl)pentanamide)를 수득하였다(117.00 mg, 56 % 수율).N- (4-fluorophenyl) -N- (prop-2-ynyl) pentaneamide (300.00 mg, 1.29 mmol) and 4-iodophenol (140.80 mg, obtained in step 3) N- (4-fluorophenyl) -N- (3- (4-hydroxyphenyl) prop-2-ynyl) pentaneamide (N- (4-fluorophenyl) -N- (3 -(4-hydroxyphenyl) prop-2-ynyl) pentanamide) was obtained (117.00 mg, 56% yield).
1H NMR (CDCl3, 500 MHz) δ 7.22 (6H, m), 6.84 (2H, d, J = 8.0 Hz), 4.65 (2H, s), 2.06 (2H, t), 1.55 (2H, m), 1.21 (2H, m), 0.79 (3H, t) 1 H NMR (CDCl 3 , 500 MHz) δ 7.22 (6H, m), 6.84 (2H, d, J = 8.0 Hz), 4.65 (2H, s), 2.06 (2H, t), 1.55 (2H, m) , 1.21 (2H, m), 0.79 (3H, t)
단계 4 : ethyl 2-(4-(3-(N-(4-Step 4: ethyl 2- (4- (3- (N- (4- 플로오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )) 페녹시Phenoxy )아세테이트 (ethyl 2-(4-(3-(N-(4-Acetate (ethyl 2- (4- (3- (N- (4- fluorophenylfluorophenyl )) pentanamidopentanamido )prop-1-ynyl)phenoxy)acetate)의 제조) prop-1-ynyl) phenoxy) acetate)
상기 단계 4에서 수득한 N-(4-플로오로페닐)-N-(3-(4-하이드록시페닐)프로프-2-이닐)펜탄아마이드(110.00 mg, 0.34 mmol)와 에틸브로모아세테이트(Ethyl bromoacetate)(0.04 ml, 0.37 mmol)을 이용하여 상기 실시예 27의 단계 1과 동일한 방법으로 ethyl 2-(4-(3-(N-(4-플로오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세테이트 (ethyl 2-(4-(3-(N-(4-fluorophenyl)pentanamido)prop-1-ynyl)phenoxy)acetate)를 수득하였다( 113.00 mg, 81 % 수율).N- (4-fluorophenyl) -N- (3- (4-hydroxyphenyl) prop-2-ynyl) pentaneamide (110.00 mg, 0.34 mmol) and ethyl bromoacetate obtained in step 4 Ethyl 2- (4- (3- (N- (4-fluorophenyl) pentaneamido) prop- in the same manner as in Step 1 of Example 27 using Ethyl bromoacetate) (0.04 ml, 0.37 mmol) 1-ynyl) phenoxy) acetate (ethyl 2- (4- (3- (N- (4-fluorophenyl) pentanamido) prop-1-ynyl) phenoxy) acetate) was obtained (113.00 mg, 81% yield).
1H NMR (CDCl3, 500 MHz) δ 7.21 (6H, m), 6.79 (2H, d, J = 9.0 Hz), 4.65 (2H, s), 4.59 (2H, s), 4.24 (2H, m), 2.01 (2H, t), 1.52 (2H, m), 1.21 (5H, m), 0.79 (3H, t) 1 H NMR (CDCl 3 , 500 MHz) δ 7.21 (6H, m), 6.79 (2H, d, J = 9.0 Hz), 4.65 (2H, s), 4.59 (2H, s), 4.24 (2H, m) , 2.01 (2H, t), 1.52 (2H, m), 1.21 (5H, m), 0.79 (3H, t)
단계 step 5 : 25: 2 -(4-(3-(N-(4--(4- (3- (N- (4- 플로오로페닐Fluorophenyl )) 펜탄아미도Pentanamido )) 프로프Prof -1--One- 이닐Inil )) 페녹시Phenoxy )) 아세틱산Acetic acid (2-(4-(3-(N-(4-fluorophenyl)pentanamido)prop-1-ynyl)phenoxy)acetic acid)의 제조Preparation of (2- (4- (3- (N- (4-fluorophenyl) pentanamido) prop-1-ynyl) phenoxy) acetic acid)
상기 단계 5에서 수득한 ethyl 2-(4-(3-(N-(4-플로오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세테이트(100.00 mg, 0.24 mmol)를 이용하여 상기 실시예 28의 단계 5와 동일한 방법으로 2-(4-(3-(N-(4-플로오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세틱산(2-(4-(3-(N-(4-fluorophenyl)pentanamido)prop-1-ynyl)phenoxy)acetic acid)를 수득하였다(25.00 mg, 27 % 수율).Using ethyl 2- (4- (3- (N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy) acetate (100.00 mg, 0.24 mmol) obtained in step 5 above In the same manner as in Step 5 of Example 28, 2- (4- (3- (N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy) acetic acid (2- (4 -(3- (N- (4-fluorophenyl) pentanamido) prop-1-ynyl) phenoxy) acetic acid) was obtained (25.00 mg, 27% yield).
1H NMR (CDCl3, 500 MHz) δ 7.41 (2H, m), 7.26 (4H, m), 6.88 (2H, d, J = 7.0 Hz), 4.66 (2H, s), 4.60 (2H, s), 2.10 (2H, t), 1.52 (2H, m), 1.23 (2H, m), 0.82 (3H, t) 1 H NMR (CDCl 3 , 500 MHz) δ 7.41 (2H, m), 7.26 (4H, m), 6.88 (2H, d, J = 7.0 Hz), 4.66 (2H, s), 4.60 (2H, s) , 2.10 (2H, t), 1.52 (2H, m), 1.23 (2H, m), 0.82 (3H, t)
[실험예]Experimental Example
실험예 1. BLT2가 발현된 세포 또는 BLT2가 발현되지 않은 세포의 준비Experimental Example 1. Preparation of cells expressing BLT2 or cells not expressing BLT2
본 실험을 위하여, BLT2가 발현되지 않은 세포 및 BLT2가 발현된 세포 (CHO-BLT2 cells)를 하기와 같은 방법으로 준비하였다. For this experiment, cells without BLT2 expression and cells with BLT2 expression (CHO-BLT2 cells) were prepared in the following manner.
CHO 세포는 한국세포주은행 (KCLB, 10061)으로부터 얻었으며, 이를 10%의 FBS (fetal bovine serum; Life technologies, Inc.), 페니실린 (50 units/mL) 및 antibiotic antimycotic solution (Life technologies, Inc.)이 포함된 RPMI 1640 medium (Invitrogen) 에서 37 ℃, 5% CO2 조건에서 배양하였다. 상기 세포를 3일간 각각 Trypsin-EDTA를 사용하여 나누어 (splitting) 성장 단계로 유지시켰으며, PBS (phosphate-buffered saline; 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 2m MKH2PO4)로 세척하고, 이 후 새로운 배지에 첨가하여 BLT2가 발현되지 않은 세포를 준비하였다. CHO cells were obtained from the Korea Cell Line Bank (KCLB, 10061), which contained 10% FBS (fetal bovine serum; Life technologies, Inc.), penicillin (50 units / mL), and antibiotic antimycotic solution (Life technologies, Inc.). It was incubated at 37 ℃, 5% CO 2 conditions in RPMI 1640 medium (Invitrogen) containing the. The cells were maintained in the growth phase by splitting with Trypsin-EDTA for 3 days, respectively, with PBS (phosphate-buffered saline; 137 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 2m MKH 2 PO 4 ) And then added to fresh medium to prepare cells without BLT2 expression.
또한, 안정된 CHO/BLT2 클론 (stable CHO/BLT2 clones)의 제조를 위해, CHO-K1 세포를 HA-tagged human BLT2를 코딩하는 pcDNA3-long form BLT2로 형질전환하고, 0.4 mg/ml의 G418 (Invitrogen, Carlsbad, CA, USA)로 선별하였다. BLT2 발현을 스크리닝하기 위해, 상기 선별된 클론을 인간-특이적 BLT2 프라이머를 사용하는 RT-PCR로 분석하였고, 대표적인 클론을 BLT2가 발현된 세포(CHO-BLT2)로 실험에 사용하였다. In addition, for the preparation of stable CHO / BLT2 clones, CHO-K1 cells were transformed with pcDNA3-long form BLT2 encoding HA-tagged human BLT2, and 0.4 mg / ml of G418 (Invitrogen). , Carlsbad, CA, USA). To screen for BLT2 expression, the selected clones were analyzed by RT-PCR using human-specific BLT2 primers, and representative clones were used in the experiments with cells expressing BLT2 (CHO-BLT2).
실험예 2. BLT2가 발현된 세포에 대한 성장 억제효과 확인 Experimental Example 2. Confirmation of growth inhibitory effect on cells expressing BLT2
상기 실시예에서 제조한 화합물의 처리에 따른 세포 생존율을 3-(4,5-디메틸티아졸-2-일)-2,5-디페닐테트라졸륨 브로마이드 (MTT) 방법으로 측정하였다.Cell viability according to the treatment of the compound prepared in the above Example was measured by the 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) method.
보다 구체적으로, 상기 실험예 1에서 준비한 1 × 104개의 BLT2가 발현되지 않은 세포 (CHO-pcDNA3.1 cells) 및 BLT2가 발현된 세포 (CHO-BLT2 cells)를 96 mm 배양 접시 (culture dish)에 분주하고 24시간 동안 세포를 배양시켰다. 이 후, 배양액을 제거하고 무혈청 RPMI 배지를 첨가하였으며. 2시간 후, 상기 실시예에서 준비한 화합물 10 μM, 대조군인 DMSO (화합물 용매) 10 μM, 양성 대조군인 LY255283 ((1-[5-ethyl-2-hydroxy-4-[[6-methyl-6-(1H-tetrazol-5-yl)heptyl]oxy] phenyl]-ethanone) (Cayman) 10 μM을 각각의 세포에 1시간 전 처리하였다. 이 후, LTB4 (300nM)를 처리한 후 24시간 동안 배양하였다. 20μL의 MTT 용액 (5 mg/mL, Sigma-Aldrich)을 각 웰에 가하고 습한 CO2 인큐베이터에서 37 ℃, 4시간 동안 배양한 후, 상층액을 제거하고, 200μL의 DMSO를 각 웰에 가해 불용성 보라색 포마잔 결정을 용해시켰다. 550 nm에서의 흡광도를 마이크로플레이트 리더(Molecular Devices, Sunnyvale, CA)를 이용하여 측정하였으며, 모든 측정은 3회 반복 수행하다.More specifically, 1 × 10 4 prepared in Experimental Example 1 Cells without BLT2 expression (CHO-pcDNA3.1 cells) and cells with BLT2 expression (CHO-BLT2 cells) were dispensed into a 96 mm culture dish and cultured for 24 hours. Thereafter, the culture was removed and serum-free RPMI medium was added. After 2 hours, 10 μM of the compound prepared in the above example, 10 μM of DMSO (compound solvent) as a control, LY255283 ((1- [5-ethyl-2-hydroxy-4-[[6-methyl-6-) as a positive control 10 μM of (1H-tetrazol-5-yl) heptyl] oxy] phenyl] -ethanone) (Cayman) was treated 1 hour before each cell, followed by incubation for 24 hours after treatment with LTB 4 (300nM). 20 μL of MTT solution (5 mg / mL, Sigma-Aldrich) was added to each well and incubated for 4 hours at 37 ° C. in a humid CO 2 incubator, then the supernatant was removed and 200 μL of DMSO was added to each well. Insoluble purple formazan crystals were dissolved The absorbance at 550 nm was measured using a microplate reader (Molecular Devices, Sunnyvale, Calif.) And all measurements were repeated three times.
그 결과, 도 1a 내지 도 1e에 나타낸 바와 같이, BLT2가 발현된 세포(CHO-BLT2 cells)에 BLT2의 리간드인 LTB4 (300nM)를 처리한 경우 (DMSO+), 에탄올을 처리한 경우 (DMSO-)와 비교하여, 세포 성장이 20%에서 35%까지 증가하였고, BLT2가 발현된 세포 (CHO-BLT2)에서, 양성 대조군인 LY255283을 전 처리한 경우, 대조군인 DMSO를 처리한 경우와 비교하여 약 90%의 세포 성장을 나타냈으며, 상기 실시예의 화합물 처리에 따른 세포 성장 억제 효과를 확인하였다. 구체적으로, 본 발명의 화합물 (LMT-692, LMT-694, LMT-696, LMT-1013)을 10μM 전 처리한 경우, 대조군인 DMSO와 비교하여 각각 88.0%, 16.7%, 56.6%, 96.3%의 세포 성장을 나타내었는바, 성장 억제효과를 확인하였다. 이와 마찬가지로 LMT-837 (65%), LMT-841 (60%), LMT-842 (70%), LMT-883 (99%), LMT-886 (99%), LMT-1016 (99%), LMT-1018 (71.6%), LMT-1019 (99%)의 화합물에서도 성장 억제효과를 확인하였다.As a result, as shown in FIGS. 1A to 1E, LTB 4 which is a ligand of BLT2 to BLT2-expressing cells (CHO-BLT2 cells) (300 nM) treated (DMSO +) and ethanol treated (DMSO-) cell growth increased from 20% to 35%, and in BLT2 expressed cells (CHO-BLT2), positive control Phosphorus LY255283 pre-treated, showed about 90% cell growth compared to the control group treated with DMSO, confirming the effect of inhibiting cell growth according to the compound treatment of the Example. Specifically, when the compound of the present invention (LMT-692, LMT-694, LMT-696, LMT-1013) 10μM pretreatment, compared to the control group DMSO of 88.0%, 16.7%, 56.6%, 96.3% respectively Cell growth was shown, confirming the growth inhibitory effect. Similarly, LMT-837 (65%), LMT-841 (60%), LMT-842 (70%), LMT-883 (99%), LMT-886 (99%), LMT-1016 (99%), LMT-1018 (71.6%), LMT-1019 (99%) also confirmed the growth inhibitory effect.
상기 실험결과는, 본 발명의 화합물 (LMT-692, LMT-696, LMT-837, LMT-841, LMT-842, LMT-883, LMT-886, LMT-1013, LMT-1016, LMT-1018, LMT-1019)은 BLT2로 유도된 세포 증식을 매우 우수한 효율로 억제할 수 있으며, 상기 화합물은 항암, 항천식 또는 다른 형태의 BLT2 관련 염증성 질환의 억제를 위한 치료제로 활용 가능한 약학적 성분 (BLT2-blocking pharmacological molecules)으로 이용될 수 있음을 의미한다.The experimental results, the compounds of the present invention (LMT-692, LMT-696, LMT-837, LMT-841, LMT-842, LMT-883, LMT-886, LMT-1013, LMT-1016, LMT-1018, LMT-1019) can inhibit BLT2-induced cell proliferation with very good efficiency, and the compound can be used as a pharmaceutical ingredient (BLT2-) as a therapeutic agent for the inhibition of anticancer, anti-asthma or other forms of BLT2-related inflammatory diseases. blocking pharmacological molecules).
실험예 3. LTB4로 유도된 BLT2 의존적인 주화성 저해효과 확인Experimental Example 3. Confirmation of BLT2 dependent chemotaxis inhibitory effect induced by LTB 4
주화성 (Chemotactic motility)은 6.5-mm 직경의 폴리카보네이트 필터 (8-μm 의 공극 크기, Corning Costar)를 구비한 Transwell 챔버를 이용하여 분석하였다. 구체적으로, 필터의 아래쪽 표면을 37 ℃에서 1시간 동안 무혈청 RPMI 1640 배지 중의 10 μg/mL 파이브로넥틴으로 코팅하였다. 다양한 양의 LTB4를 포함한 RPMI 1640 배지와 함께 건조, 코팅된 필터를 Transwell 챔버의 아래쪽 웰에 두고, 무혈청 RPMI 1640 배지에 BLT1 및 BLT2를 안정적으로 발현하는 CHO 세포를 최종적으로 2 × 104cells/100μL로 윗쪽 웰에 로딩하여 실험하였다. 저해제들의 효과를 평가할 때 세포들은 분주 전 30분 동안 각각의 저해제로 전 처리하였다. 37℃, 5% CO2에서 3시간 동안 배양한 후, 필터들을 메탄올로 3분 동안 고정시키고, 헤마톡실린 및 에오신으로 10분 동안 염색했다. 본 실험에서, 세포는 BLT2가 발현된 세포(CHO-BLT2 cells) 및 BLT1이 발현된 세포(CHO-BLT1)를 이용하였으며, 양성 대조군으로 각각 LY255283 및 U75302를, 비교 대조군으로 BLT2의 리간드인 LTB4 , (300 nM), BLT1의 리간드인 LTB4 (10nM), LPA (lysophosphatidicacid;100nM)를 이용하였다. 주화성은 광학 현미경 하에서 (배율, ×200), 필터의 아래쪽 측면 상의 세포를 계수함으로써 정량하였다. 각 분석에서 6개의 필드를 계수하였고, 각각의 샘플은 2회씩 분석하였으며, 상기 분석은 3회씩 반복 수행하였다. Chemotactic motility was analyzed using a Transwell chamber equipped with a 6.5-mm diameter polycarbonate filter (8-μm pore size, Corning Costar). Specifically, the bottom surface of the filter was coated with 10 μg / mL fibronectin in serum free RPMI 1640 medium at 37 ° C. for 1 hour. A dry, coated filter with RPMI 1640 medium containing varying amounts of LTB 4 is placed in the bottom well of the Transwell chamber and finally 2 × 10 4 cells of CHO cells stably expressing BLT1 and BLT2 in serum-free RPMI 1640 medium. Experiments were performed by loading into the upper wells at / 100 μL. When evaluating the effects of the inhibitors, the cells were pretreated with each inhibitor for 30 minutes before dispensing. After 3 hours of incubation at 37 ° C., 5% CO 2 , the filters were fixed with methanol for 3 minutes and stained with hematoxylin and eosin for 10 minutes. In this experiment, the cells used BLT2-expressing cells (CHO-BLT2 cells) and BLT1-expressing cells (CHO-BLT1), LY255283 and U75302 respectively as a positive control, LTB 4 ligand of BLT2 as a comparative control , (300 nM), LTB 4 , the ligand of BLT1 (10 nM) and LPA (lysophosphatidic acid; 100 nM) were used. Chemotaxis was quantified by counting cells on the lower side of the filter under an optical microscope (magnification, x200). Six fields were counted in each assay, each sample analyzed twice, and the assay was repeated three times.
그 결과, 도 2a 및 도 2b 및 하기 표 1에 나타낸 바와 같이, BLT2를 발현하는 세포(CHO-BLT2 cells)에서, 본 발명의 화합물 (LMT-692, LMT-696)의 농도가 증가함에 따라 (10-4, 10-3, 10-2, 10-1, 1, 10 및 102), 무 혈청 조건 하에서 CHO-BLT2 세포의 주화성이 억제됨을 확인하였으며, LMT-692 및 LMT-696의 화합물의 IC50 (50% 억제 농도)는 각각 7.566 μM 및 2.003 μM 이었다.As a result, as shown in Figures 2a and 2b and Table 1 below, in the BLT2 expressing cells (CHO-BLT2 cells), as the concentration of the compounds of the present invention (LMT-692, LMT-696) increases ( 10 -4 , 10 -3 , 10 -2 , 10 -1 , 1, 10 and 10 2 ), it was confirmed that chemotaxis of CHO-BLT2 cells under serum-free conditions, compounds of LMT-692 and LMT-696 IC 50 (50% inhibitory concentration) was 7.566 μM and 2.003 μM, respectively.
Figure PCTKR2016008069-appb-T000001
Figure PCTKR2016008069-appb-T000001
또한, 하기 표 2에 나타낸 바와 같이, BLT2를 발현하는 세포(CHO-BLT2 cells)에서, 본 발명의 화합물 LMT-1013의 농도가 증가함에 따라, 무 혈청 조건 하에서 CHO-BLT2 세포의 주화성이 억제됨을 확인하였으며, LMT-1013 화합물의 IC50 (50% 억제 농도)는 62.35 nM 이였다.In addition, as shown in Table 2 below, in the cells expressing BLT2 (CHO-BLT2 cells), as the concentration of the compound LMT-1013 of the present invention increases, chemotaxis of CHO-BLT2 cells is suppressed under serum-free conditions. The IC 50 (50% inhibitory concentration) of the LMT-1013 compound was 62.35 nM.
마찬가지로 BLT1을 발현하는 세포(CHO-BLT1 cells)에서, 본 발명의 화합물 LMT-1013의 농도가 증가함에 따라, 무 혈청 조건 하에서 CHO-BLT2 세포의 주화성이 억제됨을 확인하였으며, LMT-1013 화합물의 IC50 (50% 억제 농도)는 및 10 μM 이상이였다.Likewise, in the cells expressing BLT1 (CHO-BLT1 cells), as the concentration of the compound LMT-1013 of the present invention was increased, it was confirmed that chemotaxis of CHO-BLT2 cells was suppressed under serum-free conditions. IC 50 (50% inhibitory concentration) and was at least 10 μΜ.
Figure PCTKR2016008069-appb-T000002
Figure PCTKR2016008069-appb-T000002
또한, 도 3a 및 도 3b에 나타낸 바와 같이, BLT2가 발현된 세포 (CHO-BLT2 cells)에 BLT2의 리간드인 LTB4 (300nM)를 처리한 경우 (DMSO+), 에탄올을 처리한 경우 (DMSO-)와 비교하여, 세포 주화성이 2.4배 증가 되었으며, 양성 대조군으로 사용된 LY255283를 전처리 (10μM)한 경우, 리간드인 LTB4를 처리한 경우와 비교하여 대비 90%의 주화성을 나타냄을 확인하였다. 이와 마찬가지로 BLT1이 발현된 세포 (CHO-BLT1)에서, 리간드인 LTB4 (10nM)를 처리한 경우 (DMSO+), 에탄올을 처리한 경우 (DMSO-)와 비교하여, 세포 주화성이 2.2배 증가되었으며, 양성 대조군으로 사용된 U75302를 전처리 (10μM)한 경우, 리간드인 LTB4를 처리한 경우와 비교하여 90%의 주화성을 나타냄을 확인하였다. 다만, 본 발명의 화합물 (LMT-692, LMT-694, LMT-696)의 경우, BLT2가 발현된 세포에 10 μM 전처리한 경우, 리간드인 LTB4 처리 (DMSO+)에 비해 주화성이 각각 66%, 90%, 70.3% 억제됨을 확인한 반면, BLT1가 발현된 세포(CHO-BLT1)에서는 리간드인 LTB4 처리 (DMSO+)에 비해 주화성이 억제되지 않음을 확인하였다.In addition, as shown in FIGS. 3A and 3B, LTB 4 which is a ligand of BLT2 in BLT2-expressing cells (CHO-BLT2 cells) When (300nM) treated (DMSO +) and ethanol treated (DMSO-), the cell chemotaxis was increased by 2.4 times, and when LY255283 used as a positive control (10μM), the ligand LTB Compared to the case of 4 treated, it was confirmed that the coinability of 90%. Similarly, in the cells expressing BLT1 (CHO-BLT1), the ligand LTB 4 (10 nM) treated (DMSO +), ethanol treated compared to (DMSO-), the cell chemotaxis was increased 2.2 times, and U75302 used as a positive control pretreated (10 μM), the ligand LTB It was confirmed that 90% of chemotaxis was shown in comparison with the case where 4 was treated. However, in the case of the compound of the present invention (LMT-692, LMT-694, LMT-696), when the BLT2 expressed cells 10 μM pretreatment, LTB 4 is a ligand Compared with treatment (DMSO +), chemotaxis was 66%, 90%, and 70.3% inhibited, whereas LTB 4 , a ligand, was expressed in BLT1-expressing cells (CHO-BLT1). It was confirmed that chemotaxis was not suppressed compared to the treatment (DMSO +).
상기 결과는, BLT2가 안정하게 발현되고 있는 세포 (CHO-BLT2)에서, 주화성 활성은 LTB4 자극에 인해 증가되며, 본 발명의 화합물 (LMT-692, LMT-696, LMT-1013)은 이러한 주화성을 현저히 저해시킬 수 있는바, LTB4에 의해 유도된 BLT2-의존적 주화성을 저해시키기 위한 약학적 성분으로 이용될 수 있음을 의미한다.The above results indicate that chemotactic activity of LTB 4 in cells stably expressing BLT2 (CHO-BLT2). Increased due to stimulation, the compounds of the present invention (LMT-692, LMT-696, LMT-1013) can significantly inhibit this chemotaxis, to inhibit the BLT2-dependent chemotaxis induced by LTB 4 It can be used as a pharmaceutical ingredient.
실험예 4. LTB4와 BLT2 결합 저해효과 확인Experimental Example 4. Confirmation of the inhibitory effect of LTB 4 and BLT2 binding
LTB4와 BLT2 결합 (ligand binding affinity) 저해는 동위원소 트리튬(H3)이 표기표 LTB4([3H]LTB4, ARC)(specific activity 160.0 Ci/mmol)를 사용하여 분석하였다. 실험방법은 CHO-BLT2 세포 2 × 106 개를 100 mm 배양접시에 깔고 48시간 동안 배양한 후 다음 과정을 진행한다. 수확한 세포를 균질기(homogenizer)로 1분씩 총 5회 사용하여 세포막의 단백질들을 분리한다. 그 후 4℃에서 45,000 RPM으로 40 분간 원심분리를 진행하여 세포막의 단백질만 수확하고 이를 40 μg/45 μL 농도로 정량하였다. 동일하게 정량된 BLT2가 포함된 세포막 단백질에 각각 동일한 양의 [3H]LTB4(5 nM)를 처리하고 농도별(10-9, 10-8, 10-7, 10-6 및 10-5 M)로 화합물질을 처리하였을 때, 트리튬이 표기된 LTB4와 BLT2의 결합 억제정도를 Hidex 300sL 액체섬광계수기를 사용하여 측정하였다.LTB 4 and BLT2 binding (ligand binding affinity) inhibition was analyzed by isotope tritium (H3) using the label LTB 4 ([3H] LTB 4, ARC) (specific activity 160.0 Ci / mmol). Experimental method is to put 2 × 10 6 CHO-BLT2 cells in a 100 mm culture dish and incubate for 48 hours before proceeding. The harvested cells are used five times for one minute in a homogenizer to separate the proteins of the cell membrane. Thereafter, centrifugation was performed for 40 minutes at 45,000 RPM at 4 ° C to harvest only proteins of the cell membrane and quantitate them at a concentration of 40 μg / 45 μL. Cell membrane proteins containing equally quantified BLT2 were treated with the same amount of [3H] LTB 4 (5 nM), respectively, and at different concentrations (10 -9 , 10 -8 , 10 -7 , 10 -6 and 10 -5 M). ), The degree of inhibition of the binding of tritium-labeled LTB 4 to BLT2 was measured using a Hidex 300sL liquid scintillation counter.
그 결과, 도 4a 및 도 4b에 나타낸 바와 같이, BLT2를 발현하는 세포(CHO-BLT2 cells)에서, 본 발명의 화합물 (LMT-696 및 LMT-1013)의 농도가 증가함에 따라 (10-9, 10-8, 10-7, 10-6 및 10-5) LTB4와 BLT2의 결합이 억제됨을 확인하였으며, LMT-696 및 LMT-1013의 화합물의 IC50 (50% 결합 억제 농도)는 각각 5.6 nM 및 30.74 nM이었다.As a result, as shown in Figs. 4A and 4B, in the cells expressing BLT2 (CHO-BLT2 cells), as the concentration of the compounds of the present invention (LMT-696 and LMT-1013) increased (10 -9 , 10 -8 , 10 -7 , 10 -6 and 10 -5 ) showed that the binding of LTB 4 to BLT2 was inhibited, and the IC 50 (50% binding inhibition concentration) of the compounds of LMT-696 and LMT-1013 was 5.6, respectively. nM and 30.74 nM.
실험예 5. BLT2 억제를 통한 항암효과 확인Experimental Example 5. Confirmation of anticancer effect through inhibition of BLT2
본 발명자들은 이전 연구에서 BLT2가 유방암 세포인 MDA-MB-231과 MDA-MB-453의 세포 내 활성산소 (ROS; reactive oxygen species)의 생성과 싸이토카인인 IL-8 (interlukin-8)의 생성을 조절하여 결과적으로 암세포의 침투(invasion)와 전이(metastasis)가 조절됨을 보고한 바 있다. 따라서 유방암 세포인 MDA-MB-231과 MDA-MB-453 세포에서, 본 발명의 화합물 처리에 따른 세포 내 활성산소 생성 억제, 및 IL-8 발현 억제 등을 확인하고자 하였다.In a previous study, we studied the production of reactive oxygen species (ROS) and the cytokine IL-8 (interlukin-8) of BLT2 in breast cancer cells MDA-MB-231 and MDA-MB-453. As a result, it has been reported that the invasion and metastasis of cancer cells is regulated. Therefore, MDA-MB-231 and MDA-MB-453 cells, which are breast cancer cells, were intended to confirm the inhibition of the production of free radicals and IL-8 expression in the cells according to the compound treatment of the present invention.
5-1. 유방암 세포의 준비5-1. Preparation of Breast Cancer Cells
유방암 세포인 MDA-MB-231 세포는 한국세포주은행 (Seoul, Korea)으로부터, MDA-MB-435 세포는 J.-H. Lee (Asan Medical Center, Seoul, Korea)로부터 얻었다. 이를 10%의 FBS(fetal bovine serum; Life technologies, Inc.), 1% 페니실린 (50 units/mL), 및 antibiotic antimycotic solution (Life technologies, Inc.)이 포함된 RPMI 1640 medium (Invitrogen)에서 37℃, 5% CO2의 조건에서 배양하였다.MDA-MB-231 cells, breast cancer cells, were obtained from Korea Cell Line Bank (Seoul, Korea), and MDA-MB-435 cells were obtained from J.-H. Obtained from Lee (Asan Medical Center, Seoul, Korea). This was 37 ° C. in RPMI 1640 medium (Invitrogen) containing 10% FBS (fetal bovine serum; Life technologies, Inc.), 1% penicillin (50 units / mL), and antibiotic antimycotic solution (Life technologies, Inc.). , And cultured under the condition of 5% CO 2 .
5-2. 세포 내 활성산소 생성 억제효과 확인5-2. Confirmation of inhibitory effect on free radical production in cells
본 발명의 화합물 (LMT-696) 처리에 따른 세포 내 활성산소 (H2O2 )를 DCF 형광의 함수로 측정하였다. 구체적으로, 활성산소 (ROS)의 측정 전 2 × 105개의 세포를 60-mm 웰에서 키우고, FBS가 보충된 RPMI 1640 배지 중에서 24시간 동안 배양하였다. 본 발명의 화합물의 효과를 평가하기 위해, 세포들을 화합물 (LMT-696)로 각각 30분 동안 전 처리하였다. 세포 내 활성산소를 측정하기 위해, 세포들을 H2O2-민감성 형광물질인 H2DCFDA [MolecularProbes(Eugene,OR)](20μM)와 함께 37℃의 어둡고 습한 CO2 인큐베이터 내에서 20분 동안 배양하였다. H2DCFDA는 세포 내에서 DCF로 가수분해되어 세포 내에 존재하며, H2O2의 존재하에서 높은 형광을 나타내는 DCF로 산화되는바, 이러한 성질을 이용하여 활성산소의 양을 측정하였다. 또한, 탐지기에서의 활성산소 생성의 확인을 위하여 세포는 트립신-EDTA를 이용하여 수확하고, 페놀레드가 함유되어 있지 않은 무혈청 RPMI 1640 중에 재현탁시켰다. DCF 형광도는 FACS Calibur flow cytometer (Becton Dickinson, NJ)를 이용하여 각각 488 및 530 nm의 여기 및 방출 파장으로 측정하였다.Intracellular reactive oxygen (H 2 O 2 ) following compound (LMT-696) treatment of the present invention was measured as a function of DCF fluorescence. Specifically, before the measurement of free radicals (ROS) 2 × 10 5 Cells were grown in 60-mm wells and incubated for 24 hours in RPMI 1640 medium supplemented with FBS. To assess the effect of the compounds of the invention, the cells were pretreated with compound (LMT-696) for 30 minutes each. To measure intracellular free radicals, cells were incubated for 20 minutes in a dark, humid CO 2 incubator at 37 ° C. with H 2 O 2 -sensitive fluorescent substance H 2 DCFDA [Molecular Probes (Eugene, OR)] (20 μM). It was. H 2 DCFDA is hydrolyzed to DCF in the cell and is present in the cell, and oxidized to DCF showing high fluorescence in the presence of H 2 O 2 , and thus the amount of active oxygen was measured using this property. In addition, cells were harvested using trypsin-EDTA and resuspended in serum-free RPMI 1640 without phenol red for confirmation of free radical production in the detector. DCF fluorescence was measured with excitation and emission wavelengths of 488 and 530 nm, respectively, using a FACS Calibur flow cytometer (Becton Dickinson, NJ).
그 결과, 도 5a 및 도 5b에 나타낸 바와 같이, 본 발명의 화합물 (LMT-696)을 처리한 경우, 유방암 세포인 MDA-MB-231 세포 및 MDA-MB-435 세포에서 활성 산소의 생성이 유의적으로 억제됨을 확인하였다.As a result, as shown in FIGS. 5A and 5B, when treated with the compound of the present invention (LMT-696), production of free radicals in MDA-MB-231 cells and MDA-MB-435 cells, which are breast cancer cells, was significant. It was confirmed that it is inhibited by.
5-3. IL-8 발현 억제효과 확인5-3. IL-8 expression inhibitory effect confirmed
본 발명의 화합물 처리에 따른 IL-8의 발현을 확인하기 위해 세포에서 Easy Blue (Intron, Sungnam, Korea)로 모든 RNA를 분리하여 260 nm 흡광도에서 정량하였다. RNA (1.25 μg)를 중합효소연쇄반응(PCR) 기법으로 역전사과정을 통해 cDNA로 합성하였다. IL-8와 glyceraldehydes-3-phosphate dehydrogenase (GAPDH)에 특이적으로 결합하는 프라이머를 사용하여 발현량을 확인하였다.In order to confirm the expression of IL-8 according to the compound treatment of the present invention, all RNAs were isolated from cells by Easy Blue (Intron, Sungnam, Korea) and quantified at 260 nm absorbance. RNA (1.25 μg) was synthesized as cDNA by reverse transcription by polymerase chain reaction (PCR) technique. Expression levels were confirmed using primers that specifically bind to IL-8 and glyceraldehydes-3-phosphate dehydrogenase (GAPDH).
그 결과, 도 6a 및 도 6b에 나타낸 바와 같이, 본 발명의 화합물 (LMT-696)을 처리한 경우, 유방암 세포인 MDA-MB-231 세포 및 MDA-MB-435 세포에서 IL-8 발현량이 유의적으로 억제됨을 확인하였다.As a result, as shown in Figs. 6A and 6B, when the compound of the present invention (LMT-696) was treated, the amount of IL-8 expression was significant in MDA-MB-231 cells and MDA-MB-435 cells, which are breast cancer cells. It was confirmed that it is inhibited by.
5-4. 유방암 세포의 침투 억제효과 확인5-4. Confirmation of Inhibition Effect of Breast Cancer Cells
본 발명의 화합물 처리에 따른 유방암 세포의 침투를 측정하는데 BioCoat Matrigel Invasion Chambers (BD Biosciences, Bedford, MA)를 사용하였다. 5 × 104개의 유방암 세포를 trypsin-EDTA로 수확한 후, 0.5% 혈청의 포함된 RPMI 1640에 재현탁시키고, Matrigel inserts에 옮겼다. 5% 혈청이 포함된 RPMI 1640를 아래 chamber에 넣고 세포를 37 ℃ 에서 36 시간 배양하였다. 각 필터들을 메탄올로 3분 동안 고정시키고, 헤마톡실린 및 에오신으로 10분 동안 염색했다. 암세포의 침투성은 광학 현미경 하에서(배율 ×200), 필터의 아래쪽 측면 상의 세포수로 정량하였다. 각 분석마다 6개의 필드를 정량하였다. 각각의 샘플은 2회씩 분석하였으며, 상기분석은 3회씩 반복 수행하였다.BioCoat Matrigel Invasion Chambers (BD Biosciences, Bedford, Mass.) Was used to measure the penetration of breast cancer cells following the compound treatment of the present invention. 5 × 10 4 breast cancer cells were harvested with trypsin-EDTA, then resuspended in RPMI 1640 included in 0.5% serum and transferred to Matrigel inserts. RPMI 1640 containing 5% serum was placed in the lower chamber and the cells were incubated at 37 ° C. for 36 hours. Each filter was fixed with methanol for 3 minutes and stained with hematoxylin and eosin for 10 minutes. Permeability of cancer cells was quantified by the number of cells on the lower side of the filter under an optical microscope (magnification x 200). Six fields were quantified for each assay. Each sample was analyzed twice, and the analysis was repeated three times.
그 결과, 도 7a 및 도 7b에 나타낸 바와 같이, 본 발명의 화합물 (LMT-696)을 처리한 경우, 암세포 침투는 MDA-MB-231에서 70%, MDA-MB-435에서는 56% 억제됨을 확인하였다. As a result, as shown in Figure 7a and 7b, when treated with the compound of the present invention (LMT-696), it was confirmed that cancer cell infiltration is suppressed 70% in MDA-MB-231, 56% in MDA-MB-435 It was.
5-5. 유방암 세포의 전이 억제효과 확인5-5. Confirmation of metastasis inhibitory effect of breast cancer cells
본 발명의 화합물 처리에 따른 유방암세포의 전이실험은 고려대학교 윤리위원회에 의해 승인되었으며, 본 실험에 사용된 모든 실험동물은 고려대학교 동물관리 및 사용위원회의 승인된 지침에 따라 수행하였다. 6주령 암컷 누드 마우스 (Charles River, Wilmington, MA)에 암세포를 주입하여 암세포 전이를 확인하였다. 유방암 세포에 본 발명의 화합물(LMT-696, 10 μM), LY255283, U75302 및 DMSO를 24시간 전처리한 후 trypsin-EDTA로 수확하여 PBS에 재현탁시켜 zoletil (50 mg/kg)로 마취시킨 마우스에 복강주사로 2 × 106 유방암 세포를 주입하였다. 5일 후부터 매 5일 간격으로 3회 본 발명의 화합물 (LMT-696, 2.5 mg/kg), LY255283 (2.5 mg/kg), U75302 (0.25 mg/kg) 및 DMSO를 복강주사 하였다. 유방암세포를 주입한 시기로부터 15주 후 마우스를 해부하여 암세포 전이를 관찰하였다.The metastasis experiment of breast cancer cells according to the compound treatment of the present invention was approved by Korea University Ethics Committee, and all experimental animals used in this experiment were performed according to the approved guidelines of Korea University Animal Care and Use Committee. Cancer cells were injected into 6-week-old female nude mice (Charles River, Wilmington, Mass.) To confirm cancer cell metastasis. After 24 hours pretreatment of the compound of the present invention (LMT-696, 10 μM), LY255283, U75302 and DMSO in breast cancer cells, harvested with trypsin-EDTA, resuspended in PBS, and anesthetized with zoletil (50 mg / kg) in mice. 2 x 10 6 breast cancer cells were injected by intraperitoneal injection. Compounds of the invention (LMT-696, 2.5 mg / kg), LY255283 (2.5 mg / kg), U75302 (0.25 mg / kg) and DMSO were intraperitoneally injected three times every five days after 5 days. Mice were dissected 15 weeks after the injection of breast cancer cells to observe cancer cell metastasis.
그 결과, 도 8, 도 9a 및 도 9b에 나타낸 바와 같이, 암세포(MDA-MB-231)의 전이는 대조군과 비교하여 본 발명의 화합물 (LMT-696)을 처리에 의해 40% 억제되었고, 양성 대조군인 LY255283를 처리에 의해 36% 억제된 반면, U75302 처리에 의해서는 암세포인 MDA-MB-231의 전이가 억제되지 않음을 확인하였다.As a result, as shown in Fig. 8, 9A and 9B, metastasis of cancer cells (MDA-MB-231) was inhibited 40% by treatment of the compound of the present invention (LMT-696) compared to the control, positive The control group LY255283 was suppressed 36% by treatment, while the U75302 treatment did not inhibit the metastasis of cancer cells MDA-MB-231.
상기 결과는, 본 발명의 화합물 (LMT-696)은 암세포의 세포 내 활성산소와 IL-8 생성을 억제하고 결과적으로 암세포의 침투와 전이를 억제시킬 수 있는바, 우수한 항암효능을 가진 약학적 성분으로 이용될 수 있음을 의미한다.The result is that the compound of the present invention (LMT-696) can inhibit the production of free radicals and IL-8 in the cells of the cancer cells and consequently inhibit the penetration and metastasis of the cancer cells, a pharmaceutical component having excellent anticancer efficacy It can be used as.
실험예 6. BLT2 억제를 통한 항천식 효과 확인Experimental Example 6. Confirmation of anti-asthma effect through BLT2 inhibition
천식의 초기 반응에서는 mast cell이 중요한 역할을 하며, 외부에서 allergen이 기도를 통해 체내에 들어오면 mast cell이 활성화되어 여러 가지 싸이토카인 (interlukin-4, interlukin-13)들을 분비하게 된다. 이러한 싸이토카인에 의해 염증세포들의 유입, 점액의 생성, 기도가 수축하는 등의 현상이 나타나게 된다. 본 발명자들은 항 천식효과를 확인하기 위해 7주령(18-20g)이 된 암컷 BALB/c 마우스를 오리엔트 (Seoungnam, Korea)로부터 공급받아 실험에 사용하였으며, 천식을 유도하였다. 1 및 14일에 암컷 C57BL/6 마우스에 보조제인 알루미늄 하이드로퍼 겔 (alum)(Pierce, Rockford, IL) 2.5 mg을 20 mg의 난백 알부민(OVA)에 포함시켜 복강 내 감작시켰다. 두 번의 초기 감작 후 21, 22 및 23 일에 초음파 분무기를 사용하여 1% OVA를 마우스에 분사하였다. 본 발명의 화합물 (LMT-696)(5 ㎎/㎏), LY255283 (5 ㎎/㎏, Cayman) 또는 DMSO는 1% OVA를 분사하기 1 시간 전에 복강주사 하였다. 초기 감작 후 24일에 기도과민성 (AHR)를 측정하였고, 25일에 천식표현형인 염증성 싸이토카인 IL-4 발현 및 염증세포(호중구; neutrophil)의 유입을 관찰하기 위해 마우스를 해부하였다. LPS(Lipopolysaccharide)로 유도한 중증천식 동물모델은 0, 1, 2 및 7일에 Balb/c 마우스에 75 μg의 난백 알부민(OVA)과 1 mg의 LPS를 코에 넣어 감작시켰다. 14, 15, 21 및 22일에 50 μg의 난백 알부민(OVA)을 코에 넣어 challenge 시켰다. 본 발명의 화합물 (LMT-1013) (1, 3, 10, 30 ㎎/㎏), montelukast (10 ㎎/㎏, DRS) 또는 control buffer (10% DMA, 5% Tween 80, 85% saline)는 50 μg의 난백 알부민(OVA)을 코에 넣어 challenge하기 1 시간 전에 먹였다. 초기 감작 후 23일에 기도과민성 (AHR)을 측정하였고 24일에 중증천식표현형인 염증세포(호중구; neutrophil)의 유입을 관찰하기 위해 마우스를 해부하였다. 또한, 기도과민성 측정은 마우스에 기도수축제인 metacholine (조건에 따라 6.25 mg/ml ~ 50 mg/ml까지)을 투여한 후 수행하였다. 기도 수축제 투여는 초음파 분무기를 사용하여 3분 동안 챔버의 입구를 통해 분사하였다. 기도의 과민성은 enhanced pause를 천식현상의 지표로 사용하여 분석하였다. 기도세척액 세포 개수는 광학 현미경 하에서 (배율, ×200), 세포를 계수함으로써 정량하였다. 각 분석에서 4개의 필드를 계수하였고, 각각의 샘플은 2회씩 분석하였으며, 상기 분석은 3회씩 반복 수행하였다. In the early reaction of asthma, mast cells play an important role. When allergen enters the body through the airway, mast cells are activated to release various cytokines (interlukin-4 and interlukin-13). The cytokine causes phenomena such as influx of inflammatory cells, mucus production, and airway contraction. To confirm the anti-asthma effect, the present inventors received female BALB / c mice that were 7 weeks old (18-20 g) from Orient (Seoungnam, Korea) and used them in the experiments to induce asthma. On day 1 and 14 2.5 mg of aluminum hydropergel (alum) (Pierce, Rockford, IL), a supplement to female C57BL / 6 mice, was intraperitoneally sensitized to 20 mg of egg white albumin (OVA). Mice were sprayed with 1% OVA using an ultrasonic nebulizer on days 21, 22 and 23 after two initial sensitizations. Compounds of the invention (LMT-696) (5 mg / kg), LY255283 (5 mg / kg, Cayman) or DMSO were intraperitoneally injected 1 hour prior to 1% OVA injection. Airway hypersensitivity (AHR) was measured 24 days after initial sensitization, and mice were dissected on day 25 to observe the asthma phenotype of inflammatory cytokine IL-4 expression and the influx of inflammatory cells (neutrophils). LPS (Lipopolysaccharide) -induced severe asthma animal model was sensitized by adding 75 μg of egg white albumin (OVA) and 1 mg of LPS to Balb / c mice on days 0, 1, 2 and 7. At 14, 15, 21 and 22 days, 50 μg of egg white albumin (OVA) was added to the nose for challenge. Compounds of the invention (LMT-1013) (1, 3, 10, 30 mg / kg), montelukast (10 mg / kg, DRS) or control buffer (10% DMA, 5% Tween 80, 85% saline) were 50 μg of egg white albumin (OVA) was placed in the nose and fed 1 hour before challenge. Airway hypersensitivity (AHR) was measured on day 23 after initial sensitization, and mice were dissected on day 24 to observe the influx of inflammatory cells (neutrophils), a severe asthmatic phenotype. In addition, airway hypersensitivity measurement was performed after the administration of metacholine (up to 6.25 mg / ml to 50 mg / ml depending on conditions), which is an airway constrictor. Airway constrictor administration was sprayed through the inlet of the chamber for 3 minutes using an ultrasonic nebulizer. Airway hypersensitivity was analyzed using enhanced pause as an indicator of asthma. Airway wash cell numbers were quantified by counting cells under an optical microscope (magnification, x 200). Four fields were counted in each assay, each sample analyzed twice, and the assay was repeated three times.
또한, 도 10 및 도 11에 나타낸 바와 같이, 양성 대조군인 LY255283을 10 μM 전 처리한 경우, 기도수축제 50 mg/ml를 투여한 중증천식이 유도된 마우스에서 기도과민성이 69.2% 감소되었으며, 마우스 복강에서 분리된 세포들의 IL-4 생성이 67.2% 감소됨을 확인하였다. 나아가, 본 발명의 화합물(LMT-696)을 10 μM 전처리한 경우, 기도수축제 50 mg/ml를 투여한 중증천식이 유도된 마우스에서 기도 과민성이 70% 감소되었으며, 마우스 복강에서 분리된 세포들의 IL-4 생성이 70% 감소됨을 확인하였다.In addition, as shown in FIGS. 10 and 11, when 10 μM of the positive control LY255283 was pretreated, airway hypersensitivity was reduced by 69.2% in severe asthma-induced mice administered with 50 mg / ml of the airway contractor. It was confirmed that IL-4 production of cells isolated from the abdominal cavity was reduced by 67.2%. Furthermore, when 10 μM of the compound of the present invention (LMT-696) was pretreated, airway hypersensitivity was reduced by 70% in severe asthma-induced mice administered with 50 mg / ml of airway constrictor. It was confirmed that IL-4 production was reduced by 70%.
또한, 도 12에 나타낸 바와 같이, 천식이 유도된 마우스 (OVA+LPS)에서 천식이 유도 되지 않은 마우스 (Normal)에 비해 13배가량 기도 저항성이 증가되었으며, 본 발명의 화합물 (LMT-1013)을 1, 3, 10 및 30 mg/kg 전처리한 경우, 기도수축제 50 mg/ml를 투여한 마우스와 비교하여, 기도 과민성이 각각 48.6%, 52.9%, 83.2%, 87.3% 감소됨을 확인하였다. 반대로 비교 물질인 montelukast 10 mg/kg을 전처리한 경우, 기도수축제 50 mg/ml를 투여한 마우스와 비교하여, 기도 과민성이 64% 감소됨을 확인하였다.In addition, as shown in FIG. 12, airway resistance was increased by about 13 times compared to asthma-induced mouse (OVA + LPS) compared to non-asthma-induced mouse (Normal), and the compound of the present invention (LMT-1013) When pretreatment with 1, 3, 10 and 30 mg / kg, it was confirmed that airway hypersensitivity was reduced by 48.6%, 52.9%, 83.2%, 87.3%, respectively, compared to mice administered 50 mg / ml airway constrictor. In contrast, pretreatment with 10 mg / kg of montelukast, a comparative substance, resulted in a 64% reduction in airway hypersensitivity compared to mice administered 50 mg / ml of airway constrictor.
또한, 도 13a, 도 13b 및 도 13c에 나타낸 바와 같이, 본 발명의 화합물 (LMT-1013)을 1 및 10 mg/kg 전처리한 경우, 마우스 복강에서 유입된 전체 세포와 호중구가 감소됨을 확인하였고, 특히 면역세포인 호중구가 각각 51.6%, 90.3% 감소됨을 확인하였다. In addition, as shown in Figure 13a, Figure 13b and Figure 13c, when the compound of the present invention (LMT-1013) 1 and 10 mg / kg pretreatment, it was confirmed that the total cells and neutrophils introduced into the mouse abdominal cavity, In particular, it was confirmed that neutrophils, which are immune cells, were reduced by 51.6% and 90.3%, respectively.
또한, 도 14a 및 도 14b에 나타낸 바와 같이, 본 발명의 화합물 (LMT-1013)을 1, 3, 10, 및 30 mg/kg 전처리한 경우, 마우스 복강에서 유입된 전체 세포와 호중구가 감소됨을 확인하였고, 특히 유입된 호중구가 각각 42.2%, 48.8%, 71.8%, 88.3% 감소됨을 확인하였다. 반대로 비교 물질인 montelukast 10 mg/kg을 전처리한 경우, 마우스 복강에서 유입된 면역세포인 호중구가 감소되지 않음을 확인하였다.In addition, as shown in Figures 14a and 14b, when the compound of the present invention (LMT-1013) 1, 3, 10, and 30 mg / kg pretreatment, it was confirmed that the total cells and neutrophils introduced from the mouse abdominal cavity In particular, the inflow of neutrophils was found to decrease by 42.2%, 48.8%, 71.8%, 88.3%, respectively. In contrast, pretreatment with 10 mg / kg of montelukast, a comparative substance, did not reduce neutrophils, which are immune cells introduced from the abdominal cavity of the mouse.
상기 결과는, 본 발명의 화합물(LMT-696 및 LMT-1013)은 천식 동물모델에서 기도과민성을 억제하고 화합물(LMT-696)은 염증성 싸이토카인 IL-4 생성을 억제시키며 화합물(LMT-1013)은 복강으로의 면역세포의 유입을 억제시켜 결과적으로 천식의 증상을 완화시킬 수 있는바, 항천식 효과를 갖는 약학적 성분으로 이용될 수 있음을 의미한다.The results indicate that the compounds of the present invention (LMT-696 and LMT-1013) inhibit airway hyperresponsiveness in the asthma animal model and the compound (LMT-696) inhibits the production of inflammatory cytokines IL-4 and the compound (LMT-1013) By inhibiting the influx of immune cells into the abdominal cavity, as a result, it is possible to alleviate the symptoms of asthma, which means that it can be used as a pharmaceutical component having an anti-asthmatic effect.
전술한 본 발명의 설명은 예시를 위한 것이며, 본 발명이 속하는 기술분야의 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다.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.
본 발명은 BLT2 (Leukotriene B4 receptor 2) 억제 활성을 나타내는 신규 화합물 및 이를 포함하는 염증성 질환 예방 또는 치료용 약학적 조성물에 관한 것이다. 본 발명자들은 종래의 염증성 질환 치료 물질의 문제점인 생체 내 불안정성 및 대량 생산의 어려움을 해소하기 위하여 BTL2 억제 활성을 나타내는 신규 화합물을 규명하였으며, 상기 화합물의 우수한 암세포 사멸 증진 및 전이 억제 효과, 주화성 억제 효과, 및 항 천식 효과 등을 실험적으로 확인하였는바, 염증성 질환을 치료하기 위한 약학적 조성물로 유용하게 사용될 수 있을 것으로 기대된다.The present invention relates to a novel compound that exhibits Leukotriene B4 receptor 2 (BLT2) inhibitory activity and a pharmaceutical composition for preventing or treating an inflammatory disease comprising the same. The present inventors have identified a novel compound that exhibits BTL2 inhibitory activity in order to solve in vivo instability and difficulty in mass production, which are problems of conventional inflammatory disease treatment substances, and has excellent cancer cell death enhancement and metastasis suppression effect and chemotaxis inhibition of the compound. The effects, and anti-asthma effects have been confirmed experimentally, it is expected that it can be usefully used as a pharmaceutical composition for treating inflammatory diseases.

Claims (7)

  1. 하기 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용 가능한 염:A compound represented by formula (1) or a pharmaceutically acceptable salt thereof:
    [화학식 1][Formula 1]
    Figure PCTKR2016008069-appb-I000039
    Figure PCTKR2016008069-appb-I000039
    상기 화학식 1에서, In Chemical Formula 1,
    R1은 C1~C10의 알킬이고,R 1 is C 1 -C 10 is alkyl,
    R2는
    Figure PCTKR2016008069-appb-I000040
    ,
    Figure PCTKR2016008069-appb-I000041
    ,
    Figure PCTKR2016008069-appb-I000042
    ,
    Figure PCTKR2016008069-appb-I000043
    또는
    Figure PCTKR2016008069-appb-I000044
    이며,     R 2 is
    Figure PCTKR2016008069-appb-I000040
    ,
    Figure PCTKR2016008069-appb-I000041
    ,
    Figure PCTKR2016008069-appb-I000042
    ,
    Figure PCTKR2016008069-appb-I000043
    or
    Figure PCTKR2016008069-appb-I000044
    Is,
    Ra는
    Figure PCTKR2016008069-appb-I000045
    ,
    Figure PCTKR2016008069-appb-I000046
    ,
    Figure PCTKR2016008069-appb-I000047
    ,
    Figure PCTKR2016008069-appb-I000048
    ,
    Figure PCTKR2016008069-appb-I000049
    ,
    Figure PCTKR2016008069-appb-I000050
    ,
    Figure PCTKR2016008069-appb-I000051
    ,
    Figure PCTKR2016008069-appb-I000052
    ,
    Figure PCTKR2016008069-appb-I000053
    ,
    Figure PCTKR2016008069-appb-I000054
    ,
    Figure PCTKR2016008069-appb-I000055
    ,
    Figure PCTKR2016008069-appb-I000056
    ,
    Figure PCTKR2016008069-appb-I000057
    ,
    Figure PCTKR2016008069-appb-I000058
    ,
    Figure PCTKR2016008069-appb-I000059
    ,
    Figure PCTKR2016008069-appb-I000060
    ,
    Figure PCTKR2016008069-appb-I000061
    ,
    Figure PCTKR2016008069-appb-I000062
    ,
    Figure PCTKR2016008069-appb-I000063
    ,
    Figure PCTKR2016008069-appb-I000064
    ,
    Figure PCTKR2016008069-appb-I000065
    ,
    Figure PCTKR2016008069-appb-I000066
    또는 하이드록시이고,    R a is
    Figure PCTKR2016008069-appb-I000045
    ,
    Figure PCTKR2016008069-appb-I000046
    ,
    Figure PCTKR2016008069-appb-I000047
    ,
    Figure PCTKR2016008069-appb-I000048
    ,
    Figure PCTKR2016008069-appb-I000049
    ,
    Figure PCTKR2016008069-appb-I000050
    ,
    Figure PCTKR2016008069-appb-I000051
    ,
    Figure PCTKR2016008069-appb-I000052
    ,
    Figure PCTKR2016008069-appb-I000053
    ,
    Figure PCTKR2016008069-appb-I000054
    ,
    Figure PCTKR2016008069-appb-I000055
    ,
    Figure PCTKR2016008069-appb-I000056
    ,
    Figure PCTKR2016008069-appb-I000057
    ,
    Figure PCTKR2016008069-appb-I000058
    ,
    Figure PCTKR2016008069-appb-I000059
    ,
    Figure PCTKR2016008069-appb-I000060
    ,
    Figure PCTKR2016008069-appb-I000061
    ,
    Figure PCTKR2016008069-appb-I000062
    ,
    Figure PCTKR2016008069-appb-I000063
    ,
    Figure PCTKR2016008069-appb-I000064
    ,
    Figure PCTKR2016008069-appb-I000065
    ,
    Figure PCTKR2016008069-appb-I000066
    Or hydroxy,
    Rb는
    Figure PCTKR2016008069-appb-I000067
    ,
    Figure PCTKR2016008069-appb-I000068
    ,
    Figure PCTKR2016008069-appb-I000069
    또는
    Figure PCTKR2016008069-appb-I000070
    이고,    R b is
    Figure PCTKR2016008069-appb-I000067
    ,
    Figure PCTKR2016008069-appb-I000068
    ,
    Figure PCTKR2016008069-appb-I000069
    or
    Figure PCTKR2016008069-appb-I000070
    ego,
    RC는
    Figure PCTKR2016008069-appb-I000071
    ,
    Figure PCTKR2016008069-appb-I000072
    또는
    Figure PCTKR2016008069-appb-I000073
    이고,     R C is
    Figure PCTKR2016008069-appb-I000071
    ,
    Figure PCTKR2016008069-appb-I000072
    or
    Figure PCTKR2016008069-appb-I000073
    ego,
    Rd는 수소 또는
    Figure PCTKR2016008069-appb-I000074
    이고,    R d is hydrogen or
    Figure PCTKR2016008069-appb-I000074
    ego,
    Re는
    Figure PCTKR2016008069-appb-I000075
    또는
    Figure PCTKR2016008069-appb-I000076
    이고,    R e is
    Figure PCTKR2016008069-appb-I000075
    or
    Figure PCTKR2016008069-appb-I000076
    ego,
    R3는 수소 또는 플루오르이다.R 3 is hydrogen or fluorine.
  2. 제 1항에 있어서,The method of claim 1,
    상기 화학식 1로 표시되는 화합물은 하기 화합물들로 이루어진 군으로부터 선택되는 것을 특징으로 하는, 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용 가능한 염:The compound represented by Formula 1 is selected from the group consisting of the compound, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof:
    tert-부틸 4-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; tert -butyl 4- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) benzoyl) piperazin-1-carboxylate;
    N-페닐-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N -phenyl- N- (3- (4- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide;
    N-(3-(4-(4-메틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4- (4-methylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide;
    N-(3-(4-(4-에틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4- (4-ethylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide;
    N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4- (4-isopropylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide;
    N-(3-(4-(4-(2-하이드록시에틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4- (4- (2-hydroxyethyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide;
    N-(3-(4-(4-(사이클로프로필메틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4- (4- (cyclopropylmethyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide;
    N-(3-(4-(4-사이클로헥실피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4- (4-cyclohexylpiperazine-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide;
    N-(3-(4-(4-(사이클로헥실메틸)피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4- (4- (cyclohexylmethyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide;
    N-(3-(4-(4-아이소부틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4- (4-isobutylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide;
    N-페닐-N-(3-(4-(4-(프로프-2-이닐)피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N -phenyl- N- (3- (4- (4- (prop-2-ynyl) piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide;
    N-(3-(4-(4-시아노피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4- (4-cyanopiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide;
    tert-부틸 4-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; tert -butyl 4- (4- (3- ( N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate;
    N-(3-플루오로페닐)-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N - (3-fluorophenyl) - N - (3- (4- ( piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide;
    N-(3-플루오로페닐)-N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N - (3-fluorophenyl) - N - (3- (4- (4- isopropyl-piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide;
    tert-부틸 4-(4-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; tert -butyl 4- (4- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate;
    N-(4-플루오로페닐)-N-(3-(4-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N - (4-fluorophenyl) - N - (3- (4- ( piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide;
    N-(4-플루오로페닐)-N-(3-(4-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N - (4-fluorophenyl) - N - (3- (4- (4-isopropyl-piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide;
    N-(3-(4-(몰폴린-4-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4- (morpholin-4-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide;
    N-페닐-N-(3-(4-(피페리딘-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N -phenyl- N- (3- (4- (piperidine-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide;
    N,N-다이에틸-4-(3-(N-페닐펜탄아미도)프로프-1-이닐)벤즈아마이드; N , N -diethyl-4- (3- ( N -phenylpentaneamido) prop-1-ynyl) benzamide;
    N-페닐-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N -phenyl- N- (3- (3- (piperazin-1-carbonyl) phenyl) prop-2-ynyl) pentaneamide;
    N-(3-(3-(4-메틸피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (3- (4-methylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentanamide;
    N-(3-(3-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (3- (4-isopropylpiperazin-1-carbonyl) phenyl) prop-2-ynyl) -N -phenylpentaneamide;
    tert-부틸-4-(3-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)벤조일)피페라진-1-카복실레이트; tert -butyl-4- (3- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) benzoyl) piperazine-1-carboxylate;
    N-(4-플루오로페닐)-N-(3-(3-(피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N - (4-fluorophenyl) - N - (3- (3- ( piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide;
    N-(4-플루오로페닐)-N-(3-(3-(4-아이소프로필피페라진-1-카보닐)페닐)프로프-2-이닐)펜탄아마이드; N - (4- fluorophenyl) - N - (3- (3- (4- isopropyl-piperazine-1-carbonyl) phenyl) prop-2-ynyl) pentane amide;
    N-(3-(4-하이드록시페닐)프로프-2-이닐)-N-페닐펜탄아마이드; N- (3- (4-hydroxyphenyl) prop-2-ynyl) -N -phenylpentanamide;
    2-(4-(3-(N-페닐펜탄아미도)프로프-1-이닐)페녹시)아세트산;2- (4- (3- ( N -phenylpentaneamido) prop-1-ynyl) phenoxy) acetic acid;
    tert-부틸 4-(5-(3-((N-페닐펜탄아미도)프로프-1-인-1-일)피콜리노일)피페라진-1-카복실레이트; tert -butyl 4- (5- (3-(( N -phenylpentaneamido) prop-1-yn-1-yl) picolinoyl) piperazin-1-carboxylate;
    N-페닐-N-(3-(6-(피페라진-1-카보닐)피리딘-3-일)프로프-2-인-1-일)펜탄아마이드; N -phenyl- N- (3- (6- (piperazin-1-carbonyl) pyridin-3-yl) prop-2-yn-1-yl) pentanamide;
    N-(3-(6-아이소프로필피페라진-1-카보닐)피리딘-3-일)프로프-2-인-1일)펜탄아마이드; N- (3- (6-isopropylpiperazin-1-carbonyl) pyridin-3-yl) prop-2-yn-1 yl) pentaneamide;
    N,N-다이에틸-4-(3-(N-(3-플루오로페닐)펜타아미도)프로프-1-인-1-일)벤즈아마이드; N, N -diethyl-4- (3- ( N- (3-fluorophenyl) pentamido) prop-1-yn-1-yl) benzamide;
    N,N-다이에틸-4-(3-(N-(4-플루오로페닐)펜타아미도)프로프-1-인-1-일)벤즈아마이드; N, N -diethyl-4- (3- ( N- (4-fluorophenyl) pentamido) prop-1-yn-1-yl) benzamide;
    N-(3-(4-(N,N-다이에틸설파모일)페닐)프로프-2-이닐)-N-페닐펜타아마이드; N- (3- (4- ( N, N -diethylsulfamoyl) phenyl) prop-2-ynyl) -N -phenylpentamide;
    N-(3-(4-(N-아이소프로필설파모일)페닐)프로프-2-이닐)-N-페닐펜타아마이드; N- (3- (4- ( N -isopropylsulfamoyl) phenyl) prop-2-ynyl) -N -phenylpentamide;
    tert-부틸 4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤조에이트; tert -butyl 4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzoate;
    4-(3-(N-페닐펜탄아미도)프로-1-핀-1-일)벤조익산;4- (3- ( N -phenylpentanamido) pro-1-pin-1-yl) benzoic acid;
    N-에틸-4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아마이드; N -ethyl-4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamide;
    N-(2-(다이에틸아미노)에틸)-4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아마이드; N- (2- (diethylamino) ethyl) -4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamide;
    에틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)아세테이트;Ethyl 2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetate;
    2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)아세틱산;2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) acetic acid;
    메틸 2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)프로파노에이트;Methyl 2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) propanoate;
    2-(4-(3-(N-페닐펜탄아미도)프로프-1-인-1-일)벤즈아미도)프로피오닉산;2- (4- (3- ( N -phenylpentaneamido) prop-1-yn-1-yl) benzamido) propionic acid;
    2-(4-(3-(N-(3-플루오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세트산; 및 2- (4- (3- ( N- (3-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy) acetic acid; And
    2-(4-(3-(N-(4-플루오로페닐)펜탄아미도)프로프-1-이닐)페녹시)아세트산.2- (4- (3- ( N- (4-fluorophenyl) pentaneamido) prop-1-ynyl) phenoxy) acetic acid.
  3. 제 1항 또는 제 2항의 화합물 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는, 염증성 질환 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating inflammatory diseases, comprising the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient.
  4. 제 3항에 있어서,The method of claim 3, wherein
    상기 염증성 질환은 천식, 죽상경화증, 암, 피부가려움증, 류마티스 관절염 및 염증성 장 질환으로 이루어진 군으로부터 선택되는 것을 특징으로 하는, 조성물.Wherein said inflammatory disease is selected from the group consisting of asthma, atherosclerosis, cancer, itching of the skin, rheumatoid arthritis and inflammatory bowel disease.
  5. 제 3항에 있어서,The method of claim 3, wherein
    상기 조성물은 BLT2 (Leukotriene B4 receptor 2) 활성을 저해시키는 것을 특징으로 하는, 조성물.Said composition is characterized in that it inhibits Leukotriene B4 receptor 2 (BLT2) activity.
  6. 제 1항 또는 제 2항의 화합물 또는 이의 약학적으로 허용가능한 염을 개체에 투여하는 단계를 포함하는, 염증성 질환의 예방 또는 치료 방법.A method of preventing or treating an inflammatory disease comprising administering to a subject a compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof.
  7. 제 1항 또는 제 2항의 화합물 또는 이의 약학적으로 허용가능한 염의 염증성 질환의 예방 또는 치료 용도.Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof for the prophylaxis or treatment of an inflammatory disease.
PCT/KR2016/008069 2015-07-24 2016-07-23 Novel compound having blt inhibitory activity and composition, for preventing or treating inflammatory diseases, comprising same as active ingredient WO2017018750A1 (en)

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ES16830783T ES2834549T3 (en) 2015-07-24 2016-07-23 Novel compound that has BLT inhibitory activity and composition, to prevent or treat inflammatory diseases, which comprises the same as active principle
EP16830783.3A EP3327000B1 (en) 2015-07-24 2016-07-23 Novel compound having blt inhibitory activity and composition, for preventing or treating inflammatory diseases, comprising same as active ingredient
CN201680055514.4A CN108349874B (en) 2015-07-24 2016-07-23 Compound having BLT inhibitory activity and composition for preventing or treating inflammatory disease comprising the same as active ingredient
US15/745,337 US10179764B2 (en) 2015-07-24 2016-07-23 Compound having BLT inhibitory activity and composition, for preventing or treating inflammatory diseases, comprising same as active ingredient
US16/181,953 US10766857B2 (en) 2015-07-24 2018-11-06 Compound having BLT inhibitory activity and composition, for preventing or treating inflammatory diseases, comprising same as active ingredient

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WO2002040466A2 (en) * 2000-10-27 2002-05-23 Ortho-Mcneil Pharmaceutical, Inc. Amidoalkyl-piperidine and amidoalkyl-piperazine derivatives for treating of nervous systems disorders
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CN104045552A (en) * 2013-03-13 2014-09-17 上海先声药物研究有限公司 Medicinal compound as neuroprotective agent
KR20150080428A (en) * 2013-12-30 2015-07-09 이화여자대학교 산학협력단 Novel amide derivatives or pharmaceutically acceptable salts thereof, preparation method thereof and pharmaceutical composition for prevention or treatment of pain containing the same as an active ingredient

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WO2002040466A2 (en) * 2000-10-27 2002-05-23 Ortho-Mcneil Pharmaceutical, Inc. Amidoalkyl-piperidine and amidoalkyl-piperazine derivatives for treating of nervous systems disorders
US20060264415A1 (en) * 2005-04-01 2006-11-23 Methylgene Inc. Inhibitors of histone deacetylase
KR20130017073A (en) * 2011-08-05 2013-02-19 동국대학교 산학협력단 New biphenyl derivative or pharmaceutically acceptable salt thereof, and pharmaceutical composition for treating or preventing inflammatory diseases or autoimmune diseases containing the same as an active ingredient
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