WO2010064769A1 - 2-piperazino-4,5-double substitution-1,3-thiazole derivative and method of preparing the same, and therapeutic agent containing the same as its active ingredient for diseases related to inflammation caused by spc receptor activity - Google Patents
2-piperazino-4,5-double substitution-1,3-thiazole derivative and method of preparing the same, and therapeutic agent containing the same as its active ingredient for diseases related to inflammation caused by spc receptor activity Download PDFInfo
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- WO2010064769A1 WO2010064769A1 PCT/KR2009/004058 KR2009004058W WO2010064769A1 WO 2010064769 A1 WO2010064769 A1 WO 2010064769A1 KR 2009004058 W KR2009004058 W KR 2009004058W WO 2010064769 A1 WO2010064769 A1 WO 2010064769A1
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
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- 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
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
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- C—CHEMISTRY; METALLURGY
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
Definitions
- Sphingosylphosphorylcholine is a lysophosphatidic acid (LPA) with structurally similar spingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA). lysophospholipid), which acts as an important signaling intermediate for immune functions such as cell proliferation, migration, and inflammatory responses.
- SPC is known to be rarely present or detected in very low concentrations in normal people, but it is known to be increased thousands of times in the epidermis of atopic dermatitis patients.
- SPCs also play an important role in abnormal keratinization that is characteristic of atopic diseases [Higuchi, J.
- the present invention provides a 2-piperazino-4,5-disubstituted-1,3-thiazole derivative for the treatment of inflammation-related diseases represented by the following formula (1) and pharmaceutically acceptable salts thereof to provide.
- the present invention is a derivative of the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by Formula 1, wherein Y is a C 1 to C 5 alkyl group, phenyl group or substituted phenyl group And optical isomers of 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives separated by conventional separation methods.
- the present invention contains a 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by the formula (1) or an optical isomer thereof as an active ingredient in inflammation-related diseases caused by SPC receptor activity Its useful use is provided. That is, 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by Formula 1 or optical isomers thereof are useful as therapeutic agents for inflammatory diseases caused by SPC receptor activity by containing them as active ingredients. .
- 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives or optical isomers thereof represented by Formula 1 of the present invention contain them as active ingredients to inhibit scar formation after trauma and promote wound healing. It is useful as an external preparation for skin.
- the present invention provides 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by the following formula (1).
- R 1 and R 3 is a heteroaryl group, a phenyl group or substituted phenyl group
- R 2 is an alkenyl group of C 1 ⁇ C 10 straight or branched chain, or cyclic alkyl group, C 2 ⁇ C 10 of, C 2 and one selected from ⁇ C 10 alkynyl group, a heteroaryl group, an arylalkyl group, the group consisting of amide with a heteroaryl group, a phenyl group or substituted phenyl group of the C 5 ⁇ C 10
- X is piperacillin oscillator of 3 or A carbonyl group at position 5 (C ⁇ O) or hydrogen
- Y is a C 1 to C 5 alkyl group, a phenyl group or a substituted phenyl group or hydrogen at positions 2 or 3 of the piperazine group, wherein X and Y are At the same time except hydrogen, in the above R 1 , R 2 , R 3 and Y, the substituted
- R 1 and R 3 are heteroaryl groups, phenyl groups or substituted phenyl groups
- R 2 is C 1 straight or branched chain of ⁇ C 5, or cyclic alkyl group, C 2 ⁇ C 5 alkenyl group, C 2 ⁇ C 5 of the alkynyl group, a heteroaryl group, an arylalkyl group, from the group consisting of amide with a phenyl group or a substituted phenyl group
- Y is an alkyl group or hydrogen at C 1 -C 3 at position 2 or 3 of the piperazine group
- the substituted phenyl group is a
- the present invention provides a process for preparing 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by the formula (1).
- the solvent used in the present invention uses an organic solvent having excellent swelling effect of Wang resin or Merrifield resin.
- dichloromethane is used as the solvent.
- the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative compound of the present invention was inhibited by both ear edema and MPO activity as a result of the inflammatory reaction test induced by TPA, usually as an anti-inflammatory drug It suggests a comparable effect on the widely used hydrocortisone [ Table 4 ].
- N- ⁇ 5-benzoyl-2- [4- (2) in the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by the general formula (1) of the present invention is useful as a therapeutic agent for inflammatory diseases caused by SPC receptor activity. .
- Resin (2.00 g, 1.82 mmol) in the form of cyanocarbodiimidodithianate represented by the formula (2) was added to dimeformamide (DMF; 20 ml) and stirred at room temperature for 10 minutes, followed by 2-bromoacetophenone (PhCOCH). 2 Br; 1.08 g, 5.42 mmol) and triethylamine (Et 3 N; 0.77 mL, 5.52 mmol) were added thereto, followed by reaction at 80 ° C. for 3 hours. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with H 2 O, DMF, MeOH, DCM to obtain 4-amino-1,3-thiazole resin as a yellow solid represented by the formula (3-1).
- 4-amino-1,3-thiazole resin (1.00 g, 0.90 mmol) represented by Chemical Formula 3-1 was added to acetonitrile (MeCN; 10 ml) and stirred for 10 minutes, followed by pivaloyl chloride ((CH 3 ) 3 CCOCl; 0.93 mL, 7.50 mmol) and pyridine (0.60 mL, 7.42 mmol) were added thereto, followed by reaction at room temperature for 6 hours. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with H 2 O, DMF, MeOH and DCM to obtain 4-N-pivaloyl-1,3-thiazole resin, which is a brown solid represented by Chemical Formula 4-1.
- 4-N-pivaloyl-1,3-thiazole resin (1.00 g, 0.89 mmol) connected by a sulfanyl linker represented by Chemical Formula 4-1 was added to a dichloromethane (10 mL) solution, and stirred at room temperature for 10 minutes. Then, meta-chlorobenzoic acid (m-CPBA; 500 mg, 2.23 mmol) was added at room temperature, followed by reaction at room temperature for 12 hours. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MeOH and DCM to obtain 4-N-pivaloyl-1,3-thiazole sulfonyl resin as a yellow solid represented by the formula (5-1).
- m-CPBA meta-chlorobenzoic acid
- Step 4 addition and desorption reaction of 4-N-pivaloyl-1,3-thiazole sulfonyl resin (5-1) with substituted piperazine compound (6-1)
- SPC spinopolyphosphorylcholine
- the compounds prepared in the examples of the present invention showed an antagonistic action on the selective cell proliferative response induced by SPC to the cell proliferation response. Since excessive cell division proliferation due to inflammatory reactions occurring during the wounding process when the trauma is wound, scars form, a substance that inhibits cell proliferation can be used to prevent the formation of unnecessary scars. At the same time, agents that enhance cell proliferation can be used to promote wound healing after trauma.
- Examples 1-7 and 1-30 inhibited cell division proliferation response by SPC in a dose dependent manner. It is noteworthy that this effect has a chemical structure similar to that of SPC, and FTY720, a S1P agonist that shares some membrane receptors, did not inhibit SPC-induced cell proliferation. Therefore, the cell proliferation inhibiting action is due to the intrinsic structural activity of the compound of the present invention, and when the wound is wound, the inflammatory response that occurs during the wound healing process may inhibit excessive cell division and scarring. Can be.
- the gasket was put up and the upper chamber was assembled.
- Drug treated HUVEC cells were dispensed in 5 ⁇ 10 4 (56 ⁇ l volume) of the upper chamber and incubated for 8 hours in a 37 ° C. CO 2 incubator.
- the membrane was separated, dyed with Diff-Quik dye (Sysmex Corporation), washed with deionized water, and glued to the slide glass with the shiny side facing down.
- TPA Tetradecanoyl phorbol acetate
- TPA-induced inflammation models have been widely used to test the mechanism of action of inflammatory responses and the efficacy of inhibitors (De Young LM et al., Agents and Actions , 1989, 26, 335-341).
- TPA is a substance that causes an inflammatory response, and when applied to the ear of a subject, erythema and edema occur, and this inflammatory response can be measured by increasing activity of MPO (myeloperoxidase), an essential substance for bacterial attack of white blood cells.
- MPO myeloperoxidase
- mice 40 male 6-week-old ICR mice were prepared, and 20 ⁇ l of TPA solution (Sigma Aldrich Korea) dissolved in acetone at 125 ⁇ g / ml was applied to the left ear. After 1 hour of TPA application, 20 ⁇ l of acetone, 0.3% Example Compounds dissolved in acetone, or 0.3% hydrocortisone (Sigma Aldrich Korea) dissolved in acetone was applied to the TPA application site, and the same site 6 hours after TPA application. 20 ⁇ l was reapplied. At 24 hours after TPA application, the mice were euthanized by cervical dislocation, and the left ear was collected at a constant width to measure weight and MPO activity. Reaction inhibition rate was computed according to following formula (3) , and is shown in Table 4 .
- the compounds of Examples 1-7 and 1-30 significantly inhibited the inflammatory response induced by TPA on both measures of ear edema and MPO activity, and its potency intensity was anti-inflammatory drug. It is comparable to hydrocortisone which is widely used as.
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Abstract
Description
본 발명은 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체, 그의 제조방법 및 그를 유효성분으로 함유하는 SPC 수용체 활성에 관련된 염증질환 치료제에 관한 것으로서, 더욱 상세하게는 SPC 수용체 억제 활성을 보인 신규 화합물로서, 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체, 그로부터 분리된 광학이성질체 및 그 제조방법을 제공하고, 이를 유효성분으로 함유하는 SPC 수용체 활성으로 유발되는 염증관련 질환에 유용한 그의 용도에 관한 것이다.The present invention relates to a 2-piperazino-4,5-disubstituted-1,3-thiazole derivative, a method for preparing the same, and an agent for treating inflammatory diseases related to SPC receptor activity containing the same as an active ingredient. As a novel compound exhibiting SPC receptor inhibitory activity, it provides 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives, optical isomers isolated therefrom, and a method for preparing the same, which contains the same as an active ingredient. It relates to the use thereof useful for inflammation-related diseases caused by SPC receptor activity.
스핀고실포스포콜린(sphingosylphosphorylcholine, 이하 SPC)은 구조적으로 유사한 스핀고신-1-포스페이트(sphingosine-1-phosphate, S1P), 라이소포스포스파티딕산(lysophosphatidic acid, LPA) 등과 함께 라이소스핀고리피드(lysophospholipid) 계열 물질로 분류되며, 이러한 물질들은 세포의 증식, 이동, 염증반응 등의 면역기능에 중요한 신호전달 중간체로 작용한다. Sphingosylphosphorylcholine (SPC) is a lysophosphatidic acid (LPA) with structurally similar spingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA). lysophospholipid), which acts as an important signaling intermediate for immune functions such as cell proliferation, migration, and inflammatory responses.
SPC는 세포막의 구성성분인 스핀고미엘린(sphingomyelin)으로부터 스핀고미엘린 탈아실화 효소(sphingomyelin deacylase) 작용으로 생성된다[Higuchi K, Biochem J., 2000, 350, 747-56]. 또한, SPC는 다양한 세포의 성장 및 증식[Desai, Biochem. Biophys. Res. Commun., 1991, 181, 361-366], 혈관 신생[Boguslawski, Biochem. Biophys. Res. Commun., 2000, 272, 603-609], 및 세포 사멸[Jeon ES, Biochim Biophys Acta., 2005, 1734(1); 25-33]등에 깊이 관여하는 것으로 알려져 있다. SPC is produced by the action of spingomyelin deacylase from spingomyelin, a component of cell membranes [Higuchi K, Biochem J. , 2000 , 350 , 747-56]. In addition, SPC grows and proliferates various cells [Desai, Biochem. Biophys. Res. Commun ., 1991 , 181 , 361-366, Angiogenesis [Boguslawski, Biochem. Biophys. Res. Commun ., 2000 , 272 , 603-609, and cell death [Jeon ES, Biochim Biophys Acta ., 2005 , 1734 (1) ; 25-33] is known to be deeply involved.
SPC 관련 질환의 대표적 예는 아토피성 피부염이다. 아토피성 피부염은 각질층의 지질 함량 감소로 인하여 항균력이 감소되고 장벽 기능이 약화되어 외부 자극 물질에 대한 방어력이 떨어지게 되므로, 염증 반응이 일어나고 이에 따른 가려움증이 나타나게 된다. 또한, 가려움증은 2차 감염을 유발시키므로 과면역 반응으로 악순환하게 된다. Representative examples of SPC-related diseases are atopic dermatitis. Atopic dermatitis is due to a decrease in the lipid content of the stratum corneum, the antimicrobial activity is reduced and the barrier function is weakened, so that the defense against external stimulants is reduced, the inflammatory reaction occurs and itching appears accordingly. Itching also causes a secondary infection, resulting in a vicious cycle of hyperimmune reactions.
특히, SPC는 정상인의 경우에는 거의 존재하지 않거나 아주 낮은 농도로 감지되지만 아토피 피부염환자의 표피에서는 수천배 이상 증가되는 것으로 알려져 있다. [Higuchi K, Biochem. J., 2000, 350, 747-756] 및 [Reiko Okamoto, Journal of Lipid Research, 2003, 44, 93-102]. 이는 아토피성 피부염 환자의 각질층에서 세포간 지질(ceramide) 결핍 현상이 중요원인이 된다[Junko Hara, J. invest. Dermatol., 2000, 115, 406-413]. 또한, SPC는 아토피 질환에서 특징적으로 발생하는 비정상적 각질화 현상에도 중요한 역할을 담당한다[Higuchi, J. Lipid Res., 2001, 42, 1562-1570]. 이 같은 연구 결과들은 SPC가 아토피성 피부염의 대표적 증상 중 하나인 피부 장벽기능 장애의 직접적인 원인일 뿐만 아니라, 2차적으로 나타나는 염증 반응의 원인이 될 수 있음을 시사한다. 이를 바탕으로 SPC의 생성을 조절한다면, 피부염증질환의 새로운 치료제로 사용될 수 있다. In particular, SPC is known to be rarely present or detected in very low concentrations in normal people, but it is known to be increased thousands of times in the epidermis of atopic dermatitis patients. Higuchi K, Biochem. J. , 2000 , 350 , 747-756 and Reiko Okamoto, Journal of Lipid Research, 2003 , 44 , 93-102. This is caused by the lack of intercellular lipids in the stratum corneum of atopic dermatitis patients [Junko Hara, J. invest. Dermatol ., 2000 , 115 , 406-413. SPCs also play an important role in abnormal keratinization that is characteristic of atopic diseases [Higuchi, J. Lipid Res ., 2001 , 42, 1562-1570]. These findings suggest that SPC is not only a direct cause of skin barrier dysfunction, which is one of the major symptoms of atopic dermatitis, but also a secondary inflammatory response. Based on this, if you regulate the production of SPC, it can be used as a new treatment for dermatitis.
한편, 아토피성 피부염의 증상 중, 환자에게 고통과 삶의 질 저하를 가져오는 가려움증에 대해서는, SPC와 구조적으로 유사한 LPA(lysophosphatidic acid)가 가려움증을 유발시킨다는 연구 결과가 발표되었다[Hashimoto, Pharmacology, 2004, 72, 51-56]. 따라서, SPC 역시 체내에서 가려움증을 유발할 수 있음을 유추할 수 있으며, 아울러, 최근 SPC를 피내 주사하면 가려움증을 직접 유발시킬 수 있다는 사실이 입증된바 있다[국제특허 제06/049451호]. Among the symptoms of atopic dermatitis, the study found that lysophosphatidic acid (LPA), which is structurally similar to SPC, causes itching for the itch that causes pain and poor quality of life for patients [Hashimoto, Pharmacology , 2004]. , 72 , 51-56]. Therefore, it can be inferred that SPC can also cause itching in the body, and in recent years, it has been demonstrated that intradermal injection of SPC can directly cause itching [International Patent No. 06/049451].
본 발명자들에 의해 SPC 수용체 활성으로 유발되는 염증관련 질환 치료제로서, 신규한 티아졸계 유도체가 개시된 바 있다[대한민국 특허출원 제2007-97553호]. As a therapeutic agent for inflammation-related diseases caused by SPC receptor activity by the present inventors, a novel thiazole derivative has been disclosed (Korean Patent Application No. 2007-97553).
이에, 본 발명자들은 상기 티아졸계 유도체 중에서 SPC 수용체 억제 활성이 우수한 N-{5-벤조일-2-[4-(2-메톡시페닐)피페라진-1-일]티아졸-4-일}피발라마이드 화합물로부터 기초하여, 효율이 개선된 염증질환 치료제로 적용될 수 있는 신규 화합물을 탐색한 결과, 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 화합물을 설계하여 고체상 합성으로 합성공정을 개선하고, 본 발명의 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체를 사람유래 진피세포 및 마우스를 이용한 동물실험을 통하여 항염증 효과를 검색한 결과, 더욱 우수한 염증억제효과를 확인함으로써, 본 발명을 완성하였다.Accordingly, the present inventors have found that N- {5-benzoyl-2- [4- (2-methoxyphenyl) piperazin-1-yl] thiazol-4-yl} pi having excellent SPC receptor inhibitory activity among the thiazole derivatives. Based on the balamide compound, a novel compound that can be applied as an inflammatory disease treatment with improved efficiency was developed. As a result, a 2-piperazino-4,5-disubstituted-1,3-thiazole derivative compound was designed to provide a solid phase. The synthesis process was improved by synthesis, and the anti-inflammatory effects of the 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives of the present invention were examined through animal experiments using human-derived dermal cells and mice. As a result, the present invention was completed by confirming more excellent anti-inflammatory effect.
본 발명의 목적은 조합화학 합성기술을 이용하여 제조된, 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 및 그로부터 분리된 광학이성질체를 제공하는 것이다.It is an object of the present invention to provide 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives prepared from combinatorial chemical synthesis techniques and optical isomers separated therefrom.
본 발명의 다른 목적은 그를 유효성분으로 함유하는 SPC 수용체 활성으로 유발되는 염증관련 질환에 유용한 그의 용도를 제공하는 것이다.Another object of the present invention is to provide its use useful for inflammation-related diseases caused by SPC receptor activity containing it as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 하기 화학식 1로 표시되는 염증관련 질환 치료제용 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 및 약제학적으로 허용 가능한 그의 염을 제공한다. In order to achieve the above object, the present invention provides a 2-piperazino-4,5-disubstituted-1,3-thiazole derivative for the treatment of inflammation-related diseases represented by the following formula (1) and pharmaceutically acceptable salts thereof to provide.
화학식 1Formula 1
(상기 식에서, R1 및 R3는 헤테로아릴기, 페닐기 또는 치환된 페닐기이고, R2는 C1∼C10의 직쇄 또는 분쇄, 또는 고리형 알킬기, C2∼C10의 알케닐기, C2∼C10의 알키닐기, 헤테로아릴기, 아릴알킬기, C5∼C10의 헤테로아릴알킬기, 페닐기 또는 치환된 페닐기가 있는 아마이드로 이루어진 군에서 선택되는 어느 하나이고, X는 피페라진기의 3 또는 5번 위치의 카르보닐기(C=O) 이거나 수소를 나타내며, Y는 피페라진기의 2 또는 3번 위치의 C1∼C5의 알킬기, 페닐기 또는 치환된 페닐기 또는 수소이고, 이때, X와 Y가 동시에 수소인 경우는 제외한다. 상기 R1, R2, R3 및 Y에서, 치환된 페닐기는 할로겐원자, 니트로기, C1∼C5의 알킬기 및 C1∼C5의 알콕시기로 이루어진 군에서 선택된 어느 하나의 치환체이고, R1, R2 및 R3은 상기 치환체 1 내지 4개가 치환된 페닐기를 나타낸다.) (Wherein, R 1 and R 3 is a heteroaryl group, a phenyl group or substituted phenyl group, R 2 is an alkenyl group of C 1 ~C 10 straight or branched chain, or cyclic alkyl group, C 2 ~C 10 of, C 2 and one selected from ~C 10 alkynyl group, a heteroaryl group, an arylalkyl group, the group consisting of amide with a heteroaryl group, a phenyl group or substituted phenyl group of the C 5 ~C 10, X is piperacillin oscillator of 3 or A carbonyl group at position 5 (C═O) or hydrogen, and Y is a C 1 to C 5 alkyl group, a phenyl group or a substituted phenyl group or hydrogen at positions 2 or 3 of the piperazine group, wherein X and Y are At the same time except hydrogen, in the above R 1 , R 2 , R 3 and Y, the substituted phenyl group is selected from the group consisting of halogen atom, nitro group, C 1 -C 5 alkyl group and C 1 -C 5 alkoxy group and any selected one of the substituents, R 1, R 2 and R 3 is 1 to 4 wherein the substituents It represents a substituted phenyl group.)
더욱 바람직하게는 본 발명의 상기 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 중에서, R1 및 R3는 헤테로아릴기, 페닐기 또는 치환된 페닐기이고, R2는 C1∼C5의 직쇄 또는 분쇄, 또는 고리형 알킬기, C2∼C5의 알케닐기, C2∼C5의 알키닐기, 헤테로아릴기, 아릴알킬기, 페닐기 또는 치환된 페닐기가 있는 아마이드로 이루어진 군에서 선택되는 어느 하나이고, X는 피페라진기의 3 또는 5번 위치의 카르보닐기(C=O) 이거나 수소를 나타내며, Y는 피페라진기의 2 또는 3번 위치의 C1∼C3의 알킬기 또는 수소이고, 이때, X와 Y가 동시에 수소인 경우는 제외하고, 상기 R1, R2, R3에서, 치환된 페닐기는 할로겐원자, 니트로기, C1∼C5의 알킬기 및 C1∼C5의 알콕시기로 이루어진 군에서 선택된 어느 하나의 치환체가 1 내지 4개 치환된 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체가 염증관련 질환 치료제용으로 유용하다. More preferably, in the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative of the present invention, R 1 and R 3 are heteroaryl groups, phenyl groups or substituted phenyl groups, and R 2 is C 1 to C 5 straight or pulverized or cyclic alkyl group, C 2 to C 5 alkenyl group, C 2 to C 5 alkynyl group, heteroaryl group, arylalkyl group, phenyl group or amide with substituted phenyl group Any one selected from the group, X is a carbonyl group (C = O) at position 3 or 5 of the piperazine group or represents hydrogen, Y is a C 1 ~ C 3 alkyl group at position 2 or 3 of the piperazine group Or hydrogen, and in this case, except that X and Y are hydrogen at the same time, in R 1 , R 2 , and R 3 , the substituted phenyl group is a halogen atom, a nitro group, a C 1 -C 5 alkyl group, and C 1- is one substituent selected from the group consisting of C 5 alkoxy of 1 to 4 substituted 2-piperazino-4,5- A weighted ring-1,3-thiazole derivative is useful as a therapeutic agent for inflammatory diseases.
나아가, 본 발명은 상기 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 중, Y가 C1∼C5의 알킬기, 페닐기 또는 치환된 페닐기인 유도체로부터 통상의 분리방법에 의하여 분리된 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체의 광학이성질체를 포함한다. Furthermore, the present invention is a derivative of the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by Formula 1, wherein Y is a C 1 to C 5 alkyl group, phenyl group or substituted phenyl group And optical isomers of 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives separated by conventional separation methods.
본 발명은 조합화학 합성기술을 이용하여 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체의 제조방법을 제공한다. The present invention provides a method for preparing a 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by Chemical Formula 1 using a combination chemical synthesis technique.
또한, 본 발명은 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 또는 그 광학이성질체를 유효성분으로 함유하여 SPC 수용체 활성으로 유발되는 염증관련 질환에 유용한 그의 용도를 제공한다. 즉, 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 또는 그 광학이성질체는 그를 유효성분으로 함유하여 SPC 수용체 활성으로 유발되는 염증질환 치료제로서 유용하다. In addition, the present invention contains a 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by the formula (1) or an optical isomer thereof as an active ingredient in inflammation-related diseases caused by SPC receptor activity Its useful use is provided. That is, 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by Formula 1 or optical isomers thereof are useful as therapeutic agents for inflammatory diseases caused by SPC receptor activity by containing them as active ingredients. .
나아가, 본 발명의 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 또는 그 광학이성질체는 그를 유효성분으로 함유하여 외상 후 흉터 형성 억제 및 상처 치유 촉진용 피부외용제로서 유용하다.Furthermore, 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives or optical isomers thereof represented by Formula 1 of the present invention contain them as active ingredients to inhibit scar formation after trauma and promote wound healing. It is useful as an external preparation for skin.
또한, 본 발명의 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 또는 그 광학이성질체는 그를 유효성분으로 함유하여, SPC에 의한 세포의 주화성 이동 억제에 의한 신생혈관생성 저해제로서 유용하다.In addition, 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by the general formula (1) of the present invention or optical isomer thereof contain them as an active ingredient, and chemotactic migration of cells by SPC It is useful as an angiogenesis inhibitor by inhibition.
특히, 신생혈관생성 저해제의 유효성분으로서, 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 중, N-5-벤조일-2-[4-(2-메톡시페닐)-3-메틸피페라진-1-일]티아졸-4-일피발라미드를 함유하는 것이다.In particular, as an active ingredient of angiogenesis inhibitors, N-5-benzoyl-2- [4- () in 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by the formula (1). 2-methoxyphenyl) -3-methylpiperazin-1-yl] thiazol-4-ylpivalamide.
본 발명은 조합화학 합성기술을 이용하여 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 및 약제학적으로 허용 가능한 그의 염을 제공하고, 상기 유도체에 대하여 사람으로부터 유래된 진피세포 및 마우스를 이용한 동물실험을 통하여 항염증 효과를 검색하여 SPC 수용체에 대한 우수한 억제활성을 규명함으로써, SPC 수용체 활성에 관련된 염증질환 치료제로서의 용도를 제공하였다.The present invention provides combinatorial chemical synthesis techniques to provide 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives and pharmaceutically acceptable salts thereof, wherein the derivative is derived from human The anti-inflammatory effect was searched through animal experiments using dermal cells and mice to identify excellent inhibitory activity on SPC receptor, thereby providing a use as a therapeutic agent for inflammatory diseases related to SPC receptor activity.
이하, 본 발명을 상세히 설명하고자 한다.Hereinafter, the present invention will be described in detail.
본 발명은 하기 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체를 제공한다. The present invention provides 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by the following formula (1).
화학식 1Formula 1
(상기 식에서, R1 및 R3는 헤테로아릴기, 페닐기 또는 치환된 페닐기이고, R2는 C1∼C10의 직쇄 또는 분쇄, 또는 고리형 알킬기, C2∼C10의 알케닐기, C2∼C10의 알키닐기, 헤테로아릴기, 아릴알킬기, C5∼C10의 헤테로아릴알킬기, 페닐기 또는 치환된 페닐기가 있는 아마이드로 이루어진 군에서 선택되는 어느 하나이고, X는 피페라진기의 3 또는 5번 위치의 카르보닐기(C=O) 이거나 수소를 나타내며, Y는 피페라진기의 2 또는 3번 위치의 C1∼C5의 알킬기, 페닐기 또는 치환된 페닐기 또는 수소이고, 이때, X와 Y가 동시에 수소인 경우는 제외한다. 상기 R1, R2, R3 및 Y에서, 치환된 페닐기는 할로겐원자, 니트로기, C1∼C5의 알킬기 및 C1∼C5의 알콕시기로 이루어진 군에서 선택된 어느 하나의 치환체이고, R1, R2 및 R3은 상기 치환체 1 내지 4개가 치환된 페닐기를 나타낸다.) (Wherein, R 1 and R 3 is a heteroaryl group, a phenyl group or substituted phenyl group, R 2 is an alkenyl group of C 1 ~C 10 straight or branched chain, or cyclic alkyl group, C 2 ~C 10 of, C 2 and one selected from ~C 10 alkynyl group, a heteroaryl group, an arylalkyl group, the group consisting of amide with a heteroaryl group, a phenyl group or substituted phenyl group of the C 5 ~C 10, X is piperacillin oscillator of 3 or A carbonyl group at position 5 (C═O) or hydrogen, and Y is a C 1 to C 5 alkyl group, a phenyl group or a substituted phenyl group or hydrogen at positions 2 or 3 of the piperazine group, wherein X and Y are At the same time except hydrogen, in the above R 1 , R 2 , R 3 and Y, the substituted phenyl group is selected from the group consisting of halogen atom, nitro group, C 1 -C 5 alkyl group and C 1 -C 5 alkoxy group and any selected one of the substituents, R 1, R 2 and R 3 is 1 to 4 wherein the substituents It represents a substituted phenyl group.)
보다 바람직하게는, 상기 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 중, R1 및 R3는 헤테로아릴기, 페닐기 또는 치환된 페닐기이고, R2는 C1∼C5의 직쇄 또는 분쇄, 또는 고리형 알킬기, C2∼C5의 알케닐기, C2∼C5의 알키닐기, 헤테로아릴기, 아릴알킬기, 페닐기 또는 치환된 페닐기가 있는 아마이드로 이루어진 군에서 선택되는 어느 하나이고, X는 피페라진기의 3 또는 5번 위치의 카르보닐기(C=O) 이거나 수소를 나타내며, Y는 피페라진기의 2 또는 3번 위치의 C1∼C3의 알킬기 또는 수소이고, 이때, X와 Y가 동시에 수소인 경우는 제외하고, 상기 R1, R2, R3에서, 치환된 페닐기는 할로겐원자, 니트로기, C1∼C5의 알킬기 및 C1∼C5의 알콕시기로 이루어진 군에서 선택된 어느 하나의 치환체가 1 내지 4개 치환된 것이다. More preferably, in the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative, R 1 and R 3 are heteroaryl groups, phenyl groups or substituted phenyl groups, and R 2 is C 1 straight or branched chain of ~C 5, or cyclic alkyl group, C 2 ~C 5 alkenyl group, C 2 ~C 5 of the alkynyl group, a heteroaryl group, an arylalkyl group, from the group consisting of amide with a phenyl group or a substituted phenyl group Either selected, X is a carbonyl group (C = O) at position 3 or 5 of the piperazine group or represents hydrogen, and Y is an alkyl group or hydrogen at C 1 -C 3 at position 2 or 3 of the piperazine group In this case, except that X and Y are hydrogen at the same time, in the R 1 , R 2 , R 3 , the substituted phenyl group is a halogen atom, a nitro group, a C 1 to C 5 alkyl group and C 1 to C 5 Any one substituent selected from the group consisting of alkoxy groups of 1 to 4 is substituted.
또한, 본 발명은 상기 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 중, Y가 C1∼C5의 알킬기, 페닐기 또는 치환된 페닐기인 유도체로부터 통상의 분리방법에 의하여 분리된 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체의 광학이성체를 포함한다. In addition, the present invention is a derivative of the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by the formula (1), wherein Y is a C 1 to C 5 alkyl group, phenyl group or substituted phenyl group And optical isomers of 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives separated by conventional separation methods.
나아가, 본 발명은 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체의 약제학적으로 허용 가능한 그의 염을 포함한다. 본 발명에서의 약제학적으로 허용 가능한 염은 당해 기술분야에서 통상적인 방법에 의해 제조될 수 있는 것으로, 예를 들면, 염산, 브롬화수소, 황산, 황산수소나트륨, 인산, 탄산 등의 무기산과의 염 또는 개미산, 초산, 옥살산, 벤조산, 시트르산, 타르타르산, 글루콘산, 게스티스산, 푸마르산, 락토비온산, 살리실릭산, 또는 아세틸살리실릭산(아스피린)과 같은 유기산과 함께 약제학적으로 허용 가능한 이들의 산의 염을 형성하거나, 또는 나트륨, 칼륨 등의 알칼리금속이온과 반응하여 이들의 금속염을 형성하거나, 또는 암모늄 이온과 반응하여 또 다른 형태의 약제학적으로 허용 가능한 염을 형성할 수도 있다.Furthermore, the present invention includes pharmaceutically acceptable salts of 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by the formula (1). Pharmaceutically acceptable salts in the present invention can be prepared by conventional methods in the art, for example, salts with inorganic acids such as hydrochloric acid, hydrogen bromide, sulfuric acid, sodium hydrogen sulfate, phosphoric acid, carbonic acid, and the like. Or their pharmaceutically acceptable, together with organic acids such as formic acid, acetic acid, oxalic acid, benzoic acid, citric acid, tartaric acid, gluconic acid, gestyic acid, fumaric acid, lactobionic acid, salicylic acid, or acetylsalicylic acid (aspirin) It is also possible to form salts of acids, or to react with alkali metal ions such as sodium and potassium to form their metal salts, or to react with ammonium ions to form another form of a pharmaceutically acceptable salt.
본 발명은 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체의 제조방법을 제공한다.The present invention provides a process for preparing 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by the formula (1).
본 발명의 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체는 하기 반응식 1에 의하며, 고체상 평형 합성법을 이용한 조합화학 합성기술법으로 수행되어 제조된다. 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by the general formula (1) of the present invention are prepared according to the following Scheme 1 by a combination chemical synthesis method using a solid phase equilibrium synthesis method.
반응식 1Scheme 1
(상기 식에서, R1, R2, R3, X 및 Y는 상기에서 정의한 바와 같으며, ⓟ는 폴리스티렌-디비닐벤젠, 메타아크릴산-디메틸아크릴아미드 및 히드록실 메타아크릴산로 이루어진 군에서 선택되는 고분자 중합체 형태의 고체 지지체를 나타낸다.)Wherein R 1 , R 2 , R 3 , X and Y are as defined above, ⓟ is a polymer selected from the group consisting of polystyrene-divinylbenzene, methacrylic acid-dimethylacrylamide and hydroxyl methacrylic acid Solid support in the form of a polymer.)
더욱 상세하게는, 본 발명의 제조방법은 화학식 2로 표시되는 설파닐 링커로 연결된 티오이미노카보네이트와 2-할로아세토페논 유도체를 반응시켜, R1이 도입된 화학식 3으로 표시되는 4-아미노-1,3-티아졸 설파닐 레진을 합성하는 제1단계; More specifically, the preparation method of the present invention is a 4-amino-1 represented by Formula 3 wherein R 1 is introduced by reacting a thiaminominocarbonate linked with a sulfanyl linker represented by Formula 2 with a 2-haloacetophenone derivative. A first step of synthesizing, 3-thiazole sulfanyl resin;
화학식 3으로 표시되는 화합물의 4-아미노기에 염화 카르복실산과 중합반응시켜, R2가 도입된 화학식 4로 표시되는 4-N-아실-1,3-티아졸 설파닐 레진을 합성하는 제2단계; A second step of synthesizing the 4-N-acyl-1,3-thiazole sulfanyl resin represented by the general formula (4) wherein R 2 is introduced by polymerizing with a 4-amino group of the compound represented by the general formula (3). ;
화학식 4로 표시되는 화합물의 설파닐 링커 레진 부분을 산화반응시켜 설포닐 링커 레진으로 합성하여, 화학식 5로 표시되는 4-N-아실-1,3-티아졸 설포닐 레진을 합성하는 제3단계; 및 A third step of synthesizing the sulfanyl linker resin portion of the compound represented by the formula (4) to a sulfonyl linker resin to synthesize 4-N-acyl-1,3-thiazole sulfonyl resin represented by the formula (5) ; And
화학식 5로 표시되는 화합물과 화학식 6으로 표시되는 치환된 피페라진 화합물의 반응으로 탈리 및 첨가반응시켜, 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체를 합성하는 제4단계로 이루어진다. 2-piperazino-4,5-disubstituted-1,3-thiazole represented by Formula 1 by desorption and addition reaction by reaction of the compound represented by Formula 5 with the substituted piperazine compound represented by Formula 6 The fourth step is to synthesize the derivative.
본 발명에 따른 반응공정, 용매계의 조성 및 반응조건의 선택범위를 구체적으로 설명하면 하기와 같다.Referring to the reaction process according to the invention, the composition of the solvent system and the selection range of the reaction conditions in detail.
본 발명에서 사용되는 용매는 왕 레진(Wang resin)이나 메리필드 레진(Merrifield resin)의 팽윤효과(swelling effect)가 뛰어난 유기용매를 사용한다.The solvent used in the present invention uses an organic solvent having excellent swelling effect of Wang resin or Merrifield resin.
제1단계 반응에서는 디메틸포름아미드(DMF), 아세톤, 메탄올 또는 에탄올을 용매로 사용하며, 바람직하게는 디메틸포름아미드를 사용한다. 이 반응에서 사용되는 R1 치환체와 염기는 각각 3 당량 정도 사용하는 것이 좋으며, 바람직하기로는 2 당량 내외의 범위로 사용하는 것이 경제성이 뛰어나다. 이때, 염기로는 N,N-디이소프로필에틸아민, 트리에틸아민(Et3N), 메톡시 나트륨(NaOMe), 에톡시 나트륨(NaOEt) 등이 대표적으로 사용될 수 있다. 이때, 사용되는 R1 치환체는 상기에서 정의한 바를 포함하는 알킬할라이드류를 사용할 수 있다.In the first stage reaction, dimethylformamide (DMF), acetone, methanol or ethanol is used as the solvent, preferably dimethylformamide. It is preferable to use about 3 equivalents of the R <1> substituent and the base used in this reaction, Preferably it is excellent in economy to use in about 2 equivalents. In this case, N, N-diisopropylethylamine, triethylamine (Et 3 N), methoxy sodium (NaOMe), ethoxy sodium (NaOEt) and the like can be used as a base. In this case, the R 1 substituent used may be an alkyl halide including the bar defined above.
제2단계 반응에서는 아세토나이트라일(MeCN) 또는 디클로로메탄(CH2Cl2) 용매로 사용한다. 이 반응에서는 염기와 R2 치환체를 각각 3 당량 정도 사용하는 것이 좋으며, 바람직하기로는 2 당량 내외의 범위로 사용하는 것이 경제성이 뛰어나다. 이때, 염기로는 피리딘(pyridine), 트리에틸아민 등이 대표적으로 사용될 수 있다. 이 때 사용되는 R2 치환체는 상기에서 정의한 바를 포함하는 염화 카르복실산류를 사용할 수 있다.In the second stage reaction, acetonitrile (MeCN) or dichloromethane (CH 2 Cl 2 ) is used. In this reaction, it is preferable to use about 3 equivalents of the base and the R 2 substituent, respectively, and it is preferable to use it in the range of about 2 equivalents preferably. In this case, pyridine, triethylamine, etc. may be representatively used as the base. The R 2 substituent used at this time may be used chlorinated carboxylic acid including the bar defined above.
제3단계 반응에서는 디클로로메탄을 용매로 사용한다. 이 반응에서는 메타클로로과벤조산 또는 과산화수소는 4 당량 정도 사용하는 것이 좋으며, 바람직하기로는 2.5 당량 내외의 범위로 사용하는 것이 경제성이 뛰어나다. In the third reaction, dichloromethane is used as the solvent. In this reaction, it is preferable to use about 4 equivalents of metachloroperbenzoic acid or hydrogen peroxide. Preferably, it is excellent in economy to use it in the range of about 2.5 equivalents.
제4단계 반응에서는 다이옥산 또는 디클로로메탄 용액, R3, X, 및 Y를 포함하는 피페라진 화합물 및 염기를 사용하여 탈리반응과 첨가반응을 수행하여 본 발명이 목적하는 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체를 얻는다. 이 반응에서는 염기와 R3, X, 및 Y를 포함하는 피페라진 화합물을 각각 2.5 당량 정도 사용하는 것이 좋으며, 바람직하기로는 1.5 당량 내외의 범위로 사용하는 것이 경제성이 뛰어나다. 이때, 염기로는 피리딘, 트리에틸아민 등이 대표적으로 사용될 수 있다. 이때 사용되는 R3, X, 및 Y를 포함하는 피페라진 화합물은 상기에서 정의한 바를 포함하는 화합물을 사용할 수 있다.In the fourth step reaction, a stripping reaction and an addition reaction are carried out using a dioxane or dichloromethane solution, a piperazine compound and a base including R 3 , X, and Y, and thus 2-pipe represented by the general formula (1). Ferrazino-4,5-disubstituted-1,3-thiazole derivatives are obtained. In this reaction, it is preferable to use about 2.5 equivalents of piperazine compounds each containing a base and R 3 , X, and Y, and preferably in an amount of about 1.5 equivalents. In this case, pyridine, triethylamine, etc. may be representatively used as the base. The piperazine compound containing R 3 , X, and Y used at this time may use a compound including the bar defined above.
또한, 본 발명에 따른 상기 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체의 생성여부를 확인하기 위하여, 반응 후 최종 단계에서 상기 화학식 6으로 표시되는 화합물로부터 탈리된 목적화합물을 컬럼 크로마토그래피로 분리 정제한 다음, NMR 및 Mass 스펙트럼으로 구조를 분석 및 확인하였다. 반응중간체인 화학식 3, 4, 및 5로 표시되는 레진은 ATR-FTIR을 측정함으로써, 반응 진행 정도를 확인하였다.In addition, to confirm the production of 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by the formula (1) according to the present invention, it is represented by the formula (6) in the final step after the reaction The target compound separated from the resulting compound was separated and purified by column chromatography, and the structure was analyzed and confirmed by NMR and Mass spectra. The resins represented by Chemical Formulas 3, 4, and 5, which are reaction intermediates, confirmed the progress of the reaction by measuring ATR-FTIR.
본 발명은 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 또는 그 광학이성질체를 유효성분으로 함유하는 SPC 수용체 활성으로 유발되는 염증질환 치료제를 제공한다.The present invention provides a therapeutic agent for inflammatory diseases caused by SPC receptor activity containing 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by Formula 1 or an optical isomer thereof as an active ingredient. .
본 발명의 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체에 대한 세포 분열증식 반응을 관찰한 결과, SPC에 의해 유도되는 선택적인 세포증식 반응에 길항적(antagonistic) 작용을 확인함으로써[표 2], SPC에 의해 과도한 세포 분열 증식 반응으로 유도되는 아토피 피부염 등의 피부병에 유효하다. As a result of observing the cell division proliferation response to the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative of the present invention, it was antagonistic to the selective cell proliferation response induced by SPC. By confirming the action [ Table 2 ], it is effective for skin diseases such as atopic dermatitis induced by excessive cell division proliferation reaction by SPC.
또한, 과도한 세포 분열 증식은 외상을 입었을 때 상처가 아무는 과정에서 일어나는 염증 반응으로 인해 흉터가 형성되나, 본 발명의 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체가 세포 분열 증식을 억제하므로 불필요한 흉터의 형성을 예방하기 위한 피부외용제로 사용될 수 있다. 동시에, 외상 후 상처 치유를 촉진하기 위해 사용될 수 있다.In addition, excessive cell division proliferation causes scarring due to an inflammatory response that occurs when the wound is wound during trauma, but the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative of the present invention. Because it inhibits cell division proliferation, it can be used as an external skin preparation to prevent the formation of unnecessary scars. At the same time, it can be used to promote wound healing after trauma.
본 발명의 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 화합물은 TPA에 의해 유발된 염증 반응실험 결과, 귀 부종 및 MPO 활성이 모두 억제되었으며, 통상 항염증 약물로서 널리 사용되는 하이드로코티손에 대등한 효과를 제시한다[표 4].The 2-piperazino-4,5-disubstituted-1,3-thiazole derivative compound of the present invention was inhibited by both ear edema and MPO activity as a result of the inflammatory reaction test induced by TPA, usually as an anti-inflammatory drug It suggests a comparable effect on the widely used hydrocortisone [ Table 4 ].
따라서, 본 발명의 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체는 아토피성 피부염, 기타 질환에서 나타나는 염증, 소양증, 피부 감염증 등에 유효하며, 외상 후 흉터 형성 억제 및 상처 치유 촉진용 피부외용제로서 유용하게 사용될 수 있다.Accordingly, the 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives of the present invention are effective in atopic dermatitis, inflammation, pruritus, skin infections, and the like, and inhibit scar formation after trauma. It can be usefully used as an external skin preparation for promoting wound healing.
이에, 더욱 바람직하게는 본 발명의 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 중, N-{5-벤조일-2-[4-(2-메톡시페닐)-3-메틸피페라진-1-일]티아졸-4-일}피발라미드(실시예 1-7) 또는 N-{5-벤조일-2-[4-(2-클로로페닐)-3-메틸피페라진-1-일]티아졸-4-일}피발라미드(실시예 1-30)를 유효성분으로 함유한 경우, SPC 수용체 활성으로 유발되는 염증질환 치료제로서 유용하다. Thus, more preferably, N- {5-benzoyl-2- [4- (2) in the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by the general formula (1) of the present invention. -Methoxyphenyl) -3-methylpiperazin-1-yl] thiazol-4-yl} pivalamide (Example 1-7) or N- {5-benzoyl-2- [4- (2-chloro Phenyl) -3-methylpiperazin-1-yl] thiazol-4-yl} pivalamide (Examples 1-30) is useful as a therapeutic agent for inflammatory diseases caused by SPC receptor activity. .
나아가, 본 발명은 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 또는 그 광학이성질체를 유효성분으로 함유하는 세포의 주화성 이동 매개 증상 조절제로서의 용도를 제공한다.Furthermore, the present invention uses the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by the general formula (1) as a chemotactic migration-mediated symptom modulator of cells containing the optical isomer as an active ingredient. To provide.
상기 세포의 주화성 이동이라 함은 생체 내에서 특정한 사이토카인이나 케모카인 등의 물질에 끌려 내피세포 또는 면역세포가 이동하는 현상으로서, 면역 세포가 염증 부위로 이동되거나, 내피세포 이동에 의한 혈관신생(angiogenesis)이 일어난다.The chemotactic migration of the cells is a phenomenon in which endothelial cells or immune cells are attracted by a specific cytokine or chemokine, etc. in vivo, and immune cells are moved to an inflammation site or angiogenesis by endothelial cell migration ( angiogenesis) takes place.
이에, 본 발명의 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체는 SPC에 의해 유발되는 세포의 주화성 이동 반응에 대하여 측정한 결과, 생체 내에서 내피 세포 또는 면역세포의 이동 반응을 강력하게 억제한 결과를 얻을 수 있다[표 3].Accordingly, the 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by Formula 1 of the present invention were measured for chemotactic migration reaction of cells induced by SPC, Strongly inhibit the endothelial cell or immune cell migration reaction [ Table 3 ].
따라서, 본 발명의 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 또는 그 광학이성질체를 유효성분으로 함유한 세포의 주화성 이동 매개 증상 조절제는 내피세포 이동에 의한 혈관신생이 억제되었으며, 외부에서 침입한 항원에 대한 면역반응 증폭 과정을 조절할 수 있다. Accordingly, the chemotactic migration-mediated symptom modulator of cells containing 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by the general formula (1) or the optical isomer thereof as an active ingredient is endothelial. Angiogenesis caused by cell migration is suppressed, and the process of amplifying immune responses against externally invading antigens can be controlled.
이에, 본 발명은 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 또는 그 광학이성질체를 유효성분으로 함유하여, SPC에 의한 세포의 주화성 이동 억제에 의해 구현되는 신생혈관생성 저해제를 제공한다.Accordingly, the present invention contains 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by the formula (1) or an optical isomer thereof as an active ingredient to inhibit chemotactic migration of cells by SPC. It provides an angiogenesis inhibitor implemented by.
특히, 본 발명의 신생혈관생성 저해제는 상기 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 중, N-{5-벤조일-2-[4-(2-메톡시페닐)-3-메틸피페라진-1-일]티아졸-4-일}피발라미드(실시예 1-7)를 유효성분으로서 함유하는 것이다.In particular, the angiogenesis inhibitor of the present invention is N- {5-benzoyl-2- [4- in the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by Formula 1 above. (2-methoxyphenyl) -3-methylpiperazin-1-yl] thiazol-4-yl} pivalamide (Example 1-7) is contained as an active ingredient.
상기 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 또는 그 광학이성질체를 함유하여, SPC 수용체 활성으로 유발되는 염증관련 질환에 유용한 그의 용도에 있어서, 화학식 1로 표시되는 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체의 약제학적으로 허용 가능한 그의 염 역시 유효성분으로 함유될 수 있다. In the use thereof useful for inflammation-related diseases caused by SPC receptor activity, containing 2-piperazino-4,5-disubstituted-1,3-thiazole derivative represented by Formula 1 or an optical isomer thereof, Pharmaceutically acceptable salts of 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives represented by Formula 1 may also be included as active ingredients.
본 발명의 약제 조성물은 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체, 또는 그의 약제학적으로 허용 가능한 이들의 염 또는 그 광학이성질체에 통상의 무독성 약제학적으로 허용 가능한 담체, 보강제 및 부형제 등을 첨가하여 약제학적 분야에서 통상적인 제제, 예를 들면 정제, 캅셀제, 트로키제, 액제, 현탁제 등의 경구투여용 제제 또는 비경구투여용 제제로 제제화할 수 있다. Pharmaceutical compositions of the present invention are conventionally non-toxic pharmaceutically acceptable to 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives, or pharmaceutically acceptable salts thereof or optical isomers thereof. Carriers, adjuvant and excipients may be added to formulate into conventional formulations in the pharmaceutical art, for example, oral or parenteral formulations such as tablets, capsules, troches, solutions, suspensions and the like.
또한, 본 발명에 따른 화합물의 인체에 대한 투여용량은 환자의 나이, 몸무게, 성별, 투여형태, 건강상태 및 질환정도에 따라 달라질 수 있으며, 몸무게가 70kg인 성인환자를 기준으로 할 때 일반적으로 0.01∼1,000 mg/일이며, 의사 또는 약사의 판단에 따라 일정 시간간격으로 1일 1회 내지 수회로 분할 투여할 수도 있다.In addition, the dosage of the compound according to the present invention to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and is generally 0.01 based on an adult patient having a weight of 70 kg. It is -1000 mg / day, and may be divided once or several times a day at regular time intervals according to the judgment of a doctor or a pharmacist.
이하, 본 발명을 실시예에 의하여 상세히 설명한다.Hereinafter, the present invention will be described in detail by way of examples.
하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 범위가 하기 실시예에 한정되는 것은 아니다.The following examples are merely illustrative of the present invention, but the scope of the present invention is not limited to the following examples.
<실시예 1> 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체(화학식 1-1)의 합성Example 1 Synthesis of 2-piperazino-4,5-disubstituted-1,3-thiazole derivative (Formula 1-1)
단계 1: 칼륨 시아노카르보이미도다이티아네이트 레진(2)의 4-아미노-1,3-티아졸화 반응Step 1: 4-amino-1,3-thiazoleation reaction of potassium cyanocarbodiidodithiate resin (2)
화학식 2로 표시되는 시아노카르보이미도다이티아네이트 형태의 레진(2.00 g, 1.82 mmol)을 디메포름아마이드(DMF; 20㎖)에 넣고 상온에서 10분간 교반한 후, 2-브로모아세토페논(PhCOCH2Br; 1.08g, 5.42mmol)과 트리에틸아민(Et3N; 0.77㎖, 5.52 mmol)을 가한 후, 80℃에서 3시간 동안 흔들어 반응하였다. 반응종료 후, 반응혼합물을 여과하고 H2O, DMF, MeOH, DCM으로 반복 세척하여 화학식 3-1로 표시되는 노란색 고체인 4-아미노-1,3-티아졸 레진을 얻었다.Resin (2.00 g, 1.82 mmol) in the form of cyanocarbodiimidodithianate represented by the formula (2) was added to dimeformamide (DMF; 20 ml) and stirred at room temperature for 10 minutes, followed by 2-bromoacetophenone (PhCOCH). 2 Br; 1.08 g, 5.42 mmol) and triethylamine (Et 3 N; 0.77 mL, 5.52 mmol) were added thereto, followed by reaction at 80 ° C. for 3 hours. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with H 2 O, DMF, MeOH, DCM to obtain 4-amino-1,3-thiazole resin as a yellow solid represented by the formula (3-1).
단계 2: 4-아미노-1,3-티아졸 레진(3-1)의 N-피발로일화 반응Step 2: N-pivaloylation of 4-amino-1,3-thiazole resin (3-1)
화학식 3-1로 표시되는 4-아미노-1,3-티아졸 레진(1.00g, 0.90 mmol)을 아세토나이트릴(MeCN; 10㎖)에 넣고 10분간 교반한 후, 피발로일 클로라이드((CH3)3CCOCl; 0.93㎖, 7.50 mmol)과 피리딘(pyridine; 0.60㎖, 7.42 mmol)을 가한 후, 상온에서 6시간 동안 흔들어 반응하였다. 반응종료 후, 반응혼합물을 여과하고 H2O, DMF, MeOH 및 DCM으로 반복 세척하여 화학식 4-1로 표시되는 갈색고체인 4-N-피발로일-1,3-티아졸 레진을 얻었다.4-amino-1,3-thiazole resin (1.00 g, 0.90 mmol) represented by Chemical Formula 3-1 was added to acetonitrile (MeCN; 10 ml) and stirred for 10 minutes, followed by pivaloyl chloride ((CH 3 ) 3 CCOCl; 0.93 mL, 7.50 mmol) and pyridine (0.60 mL, 7.42 mmol) were added thereto, followed by reaction at room temperature for 6 hours. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with H 2 O, DMF, MeOH and DCM to obtain 4-N-pivaloyl-1,3-thiazole resin, which is a brown solid represented by Chemical Formula 4-1.
단계 3: 4-N-피발로일-1,3-티아졸 레진(4-1)의 산화 반응Step 3: Oxidation of 4-N-pivaloyl-1,3-thiazole Resin (4-1)
화학식 4-1로 표시되는 설파닐 링커로 연결된 4-N-피발로일-1,3-티아졸 레진(1.00g, 0.89 mmol)을 디클로로메탄(10㎖)용액에 넣고 상온에서 10분간 교반한 후, 메타-클로로벤조산(m-CPBA; 500mg, 2.23 mmol)을 상온에서 가한 다음, 상온에서 12시간 흔들어 반응하였다. 반응종료 후, 반응혼합물을 여과하고 DMF, MeOH 및 DCM으로 반복 세척하여 화학식 5-1로 표시되는 노란색 고체인 4-N-피발로일-1,3-티아졸 설포닐 레진을 얻었다.4-N-pivaloyl-1,3-thiazole resin (1.00 g, 0.89 mmol) connected by a sulfanyl linker represented by Chemical Formula 4-1 was added to a dichloromethane (10 mL) solution, and stirred at room temperature for 10 minutes. Then, meta-chlorobenzoic acid (m-CPBA; 500 mg, 2.23 mmol) was added at room temperature, followed by reaction at room temperature for 12 hours. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MeOH and DCM to obtain 4-N-pivaloyl-1,3-thiazole sulfonyl resin as a yellow solid represented by the formula (5-1).
단계 4: 4-N-피발로일-1,3-티아졸 설포닐 레진(5-1)과 치환된 피페라진 화합물(6-1)의 첨가 반응 및 탈리반응Step 4: addition and desorption reaction of 4-N-pivaloyl-1,3-thiazole sulfonyl resin (5-1) with substituted piperazine compound (6-1)
화학식 5-1로 표시되는 4-N-피발로일-1,3-티아졸 설포닐 레진(200mg, 0.17 mmol)을 디옥산(dioxane; 5㎖) 용액에 넣고 상온에서 10분간 교반한 후, 화학식 6-1로 표시되는 치환된 피페라진 화합물(64mg, 0.31 mmol)과 트리에틸아민(0.056㎖, 0.40 mmol)을 가한 후, 상온에서 12시간 동안 흔들어 반응하였다. 반응종료 후, 반응혼합물을 여과하고 여과물을 디클로로메탄과 메탈알코올로 반복세척한 후, 여과액을 합하여 감압농축하였다. 농축된 혼합물을 헥산/에틸아세테이트(1/1, v/v)의 혼합 용매 하에서 실리카겔상의 컬럼 크로마토그래피로 분리 정제하여 화학식 1-1로 표시되는 오일(26mg, 수율 36%; 레진 2-1로부터 4 단계까지의 총 수율; 레진 2-1의 로딩용량=0.90 mmol/g)을 얻었다. 4-N-pivaloyl-1,3-thiazole sulfonyl resin (200 mg, 0.17 mmol) represented by Chemical Formula 5-1 was added to a dioxane (5 ml) solution, and stirred at room temperature for 10 minutes. Substituted piperazine compound represented by the formula (6-1) (64mg, 0.31 mmol) and triethylamine (0.056ml, 0.40 mmol) was added, and the mixture was shaken at room temperature for 12 hours. After completion of the reaction, the reaction mixture was filtered and the filtrate was repeatedly washed with dichloromethane and metal alcohol, and the filtrates were combined and concentrated under reduced pressure. The concentrated mixture was separated and purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (1/1, v / v) to obtain an oil represented by the formula (I-1) (26 mg, yield 36%; from resin 2-1 Total yield up to step 4; loading capacity of resin 2-1 = 0.90 mmol / g) was obtained.
화합물 1-1: 1H-NMR δ 1.39 (s, 9H), 3.80 (s, 3H), 3.83 (m, 2H), 4.22 (m, 2H), 4.26 (s, 2H), 6.93 (d, J = 9.0 Hz, 2H), 7.18 (d, J = 9.0 Hz, 2H), 7.45-7.48 (m, 2H), 7.53 (m, 1H), 7.77-7.79 (m, 2H), 12.14 (s, 1H); m/z ([M+1]+) 493.Compound 1-1: 1 H-NMR δ 1.39 (s, 9H), 3.80 (s, 3H), 3.83 (m, 2H), 4.22 (m, 2H), 4.26 (s, 2H), 6.93 (d, J = 9.0 Hz, 2H), 7.18 (d, J = 9.0 Hz, 2H), 7.45-7.48 (m, 2H), 7.53 (m, 1H), 7.77-7.79 (m, 2H), 12.14 (s, 1H) ; m / z ([M + 1] + ) 493.
<실시예 2∼30> 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체의 합성 Examples 2-30 Synthesis of 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives
하기 화학식 1로 표시되는 화합물에서, R1, R2, R3, X, 및 Y를 하기 표 1에 기재된 바와 같이 실시하는 것을 제외하고는, 상기 실시예 1과 동일한 방법으로 수행하여 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체를 합성하였다. 또한, 합성된 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체의 분석결과를 표 1에 기재하였다. In the compound represented by Formula 1, 2 -P was carried out in the same manner as in Example 1, except that R 1 , R 2 , R 3 , X, and Y were carried out as described in Table 1 below . Ferrazino-4,5-disubstituted-1,3-thiazole derivatives were synthesized. In addition, the analysis results of the synthesized 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives are shown in Table 1.
화학식 1Formula 1
<실험예 1> 세포주 분열증식 반응 조절 Experimental Example 1 Control of Cell Line Division Proliferation Response
스핀고실포스포릴콜린(SPC)을 세포에 처리하면, 과도한 세포 분열 증식 반응을 유도하여, 아토피피부염 등의 피부병의 병리적 증상을 유발할 수 있다[Desai, Biochem. Biophys. Res. Commun., 1991, 181, 361-366]. 이에, 상기 실시예에서 제조된 화합물들에 대하여, SPC에 의해 유발되는 세포의 분열증식 반응에 대한 효과를 시험하였다. Treatment of spinopolyphosphorylcholine (SPC) with cells can induce excessive cell division and proliferative responses, leading to pathological symptoms of skin diseases such as atopic dermatitis [Desai, Biochem. Biophys. Res. Commun. , 1991 , 181 , 361-366. Thus, for the compounds prepared in the above example, the effect on the proliferative response of cells induced by SPC was tested.
일차적으로 NIH 3T3 세포(American Type Culture Collection, Manassas, VA, USA) 1×105개 (보통 1×104∼106개)를 배양 플레이트에 분주하여 배양한 후, 24시간 동안 우혈청이 포함되지 않은 RPMI 배지에서 배양하여 혈청을 고갈(serum starvation)시켰다. 여기에, 상기 실시예에서 제조된 화합물 및 대조 화합물로서, 스핀고신-1-포스페이트(sphingosine-1-phosphate; S1P)의 작용제(agonist)인 FTY720을 0.001 μM, 0.01 μM, 0.1 μM, 및 1 μM으로 처리하여 30분간 배양한 후, SPC(Biomol, Plymouth Meeting, PA, USA)를 7 μM 농도로 첨가하여 24시간 동안 37℃에서 배양하였다. 세포 증식량을 세포분열 때 복제되는 DNA 가닥에 삽입되는 [3H]-티미딘(thymidine) 반응법을 이용하여 측정하고[Beales IL, Life Sci., 2004, 75, 83-95] 증식율(%)을 하기 수학식 1에 의해 산출하여, 그 결과를 표 2에 나타내었다. Primarily in NIH 3T3 cells (American Type Culture Collection, Manassas, VA, USA), 1 × 10 5 pieces (usually 1 × 10 4 ~10 6 pieces) and then seeded in culture plates incubated, containing the bovine serum for 24 hours Serum was depleted (serum starvation) by culturing in RPMI medium. Herein, as the compound prepared in the above examples and the control compound, FTY720, which is an agonist of spingosine-1-phosphate (S1P), is 0.001 μM, 0.01 μM, 0.1 μM, and 1 μM. After incubation for 30 minutes, SPC (Biomol, Plymouth Meeting, PA, USA) was added to 7 μM concentration and incubated at 37 ℃ for 24 hours. Cell proliferation was measured using the [3H] -thymidine reaction method inserted into the DNA strand replicated during cell division [Beales IL, Life Sci ., 2004, 75, 83-95]. Was calculated by the following Equation 1 , and the results are shown in Table 2 .
수학식 1
상기 표 2에서 보는 바와 같이, 본 발명의 실시예에서 제조된 화합물은 세포 분열증식 반응에 대해 SPC에 의해 유도되는 선택적인 세포 증식 반응에 길항적 (antagonistic) 작용을 나타내었다. 외상을 입었을 때 상처가 아무는 과정에서 일어나는 염증 반응으로 인한 과도한 세포 분열 증식이 흉터를 형성시키므로, 세포 분열증식을 억제하는 물질은 불필요한 흉터의 형성을 예방하기 위한 용도로 사용될 수 있다. 동시에, 세포 분열증식을 증대시키는 물질은 외상 후 상처 치유를 촉진하기 위해 사용될 수 있다.As shown in Table 2, the compounds prepared in the examples of the present invention showed an antagonistic action on the selective cell proliferative response induced by SPC to the cell proliferation response. Since excessive cell division proliferation due to inflammatory reactions occurring during the wounding process when the trauma is wound, scars form, a substance that inhibits cell proliferation can be used to prevent the formation of unnecessary scars. At the same time, agents that enhance cell proliferation can be used to promote wound healing after trauma.
특히, 실시예 1-7 및 1-30은 SPC에 의한 세포 분열증식 반응을 용량 의존적으로 억제하였다. 특기할 만한 점은 이러한 효과가 SPC와 유사한 화학 구조를 가지며, 일부 막수용체를 공유하는 S1P 작용제인 FTY720에 의해서는 SPC에 의한 세포 분열증식에 대한 억제하는 결과가 나타나지 않았다. 따라서, 세포 분열증식 억제 작용은 본 발명의 화합물만의 고유한 구조 활성에 의한 것이며, 외상을 입었을 때 상처가 아무는 과정에서 일어나는 염증 반응으로 인해 과도한 세포 분열증식이 일어나 흉터가 형성되는 현상을 억제할 수 있다. In particular, Examples 1-7 and 1-30 inhibited cell division proliferation response by SPC in a dose dependent manner. It is noteworthy that this effect has a chemical structure similar to that of SPC, and FTY720, a S1P agonist that shares some membrane receptors, did not inhibit SPC-induced cell proliferation. Therefore, the cell proliferation inhibiting action is due to the intrinsic structural activity of the compound of the present invention, and when the wound is wound, the inflammatory response that occurs during the wound healing process may inhibit excessive cell division and scarring. Can be.
<실험예 2> SPC에 의해 유발된 주화성 세포이동반응에 대한 억제 효과Experimental Example 2 Inhibitory Effect on Chemotactic Cell Migration Response Induced by SPC
최근 SPC가 혈관내피세포 성장인자(VEGF)와 유사하게 주화성 세포 이동에도 중요한 역할을 수행한다는 연구 결과가 보고되었다[Boguslawski 등, Biochem Biophys Res Commun., 2000, 272, 603-609]. 생체 내에서 특정한 사이토카인이나 케모카인 등의 물질에 끌려 세포가 이동하는 현상은 면역세포가 염증 부위로 이동하거나, 내피세포 이동에 의한 혈관신생(angiogenesis) 같은 과정에서 핵심적인 단계이다. 이에 SPC에 의해 유발되는 세포의 주화성 이동 반응에 대하여, 상기 실시예에서 제조된 화합물들의 효과를 보이든 챔버(boyden chamber) 분석으로 시험하였다.Recently, studies have shown that SPC plays an important role in chemotactic cell migration similarly to vascular endothelial growth factor (VEGF) (Boguslawski et al ., Biochem Biophys Res Commun ., 2000, 272, 603-609). The movement of cells by attracting certain cytokines or chemokines, etc. in vivo is a key step in the process of immune cells moving to inflammation sites or angiogenesis by endothelial cell migration. In response to chemotactic migration of cells induced by SPC, the effects of the compounds prepared in the above examples were tested by a Boyden chamber assay.
먼저, 8㎛ 포어를 가진 25×80mm 폴리카보네이트 막(polycarbonate membrane; Neuro Probe, Inc.)을 0.01% 젤라틴, 0.1% 아세트산 용액에 담가 하룻밤 동안 코팅한 다음 실온에서 자연 건조시켜 준비하였다. First, a 25 × 80 mm polycarbonate membrane (Neuro Probe, Inc.) having an 8 μm pore was immersed in a 0.01% gelatin, 0.1% acetic acid solution and coated overnight, followed by natural drying at room temperature.
한편, 2% 우태아 혈청을 포함한 완전 EBM-2 배지에서 배양한 HUVEC(Human umbilical vein endothelial cell)을 4시간 동안 우혈청이 포함되지 않은 EBM-2 배지(Cambrex, 카탈로그 넘버 CC-3121)에 배양하여 혈청을 고갈(serum starvation)시킨 후, 트립신/EDTA 용액으로 수확하였다. HUVEC 세포를 0.1% 우혈청 알부민이 포함된 EBM-2 배지에 현탁시킨 후, 실리콘 코팅된 에펜도르프 튜브에 분주하여 실험 약물로서 실시예 1-7을 0, 0.1, 1 내지 10 μg/㎖의 양으로 가하여 37℃에서 30분간 처리하였다. 보이든 챔버의 아랫쪽 챔버의 각 웰에 10 μM SPC를 포함한 EBM-2 배지 또는 포함하지 않은 EBM-2 배지를 27 μL씩 분주하고, 젤라틴 코팅해 준비한 막을 윤기 있는 면이 아래로 향하게 올려놓고 그 위에 개스킷 (gasket)을 올려놓은 다음 위쪽 챔버를 조립하였다. 위쪽 챔버에 약물처리한 HUVEC 세포를 5×104(부피 56㎕)씩 분주하고 37℃ CO2 배양기에서 8시간 배양하였다. 막을 분리해 내어 Diff-Quik 염색약(Sysmex Corporation)으로 염색하고 탈이온수로 세척한 다음, 윤기 있는 면이 아래로 향하도록 슬라이드 글라스에 붙였다. 막 위쪽에 붙어있는 세포들을 킴와이퍼나 면봉으로 조심스럽게 닦아낸 다음, 각 웰당 임의로 5 필드씩 사진을 찍어서(배율 200배), 세포 수를 측정하였다. 반응 억제율은 하기의 수학식 2에 따라 산출하고, 그 결과를 표 3에 나타내었다. Meanwhile, human umbilical vein endothelial cells (HUVEC) cultured in complete EBM-2 medium containing 2% fetal bovine serum were cultured in EBM-2 medium (Cambrex, catalog number CC-3121) containing no serum for 4 hours. The serum was depleted (serum starvation) and then harvested with trypsin / EDTA solution. HUVEC cells were suspended in EBM-2 medium containing 0.1% bovine serum albumin and then dispensed into silicone coated Eppendorf tubes to give Examples 1-7 as 0, 0.1, 1 to 10 μg / ml as experimental drug. The solution was added at 37 ° C. for 30 minutes. Dispense 27 μL of EBM-2 medium or 10 μM SPC or EBM-2 medium into each well of the lower chamber of the Boyden chamber and place the membrane prepared with gelatinized coating with the glossy side facing down. The gasket was put up and the upper chamber was assembled. Drug treated HUVEC cells were dispensed in 5 × 10 4 (56 μl volume) of the upper chamber and incubated for 8 hours in a 37 ° C. CO 2 incubator. The membrane was separated, dyed with Diff-Quik dye (Sysmex Corporation), washed with deionized water, and glued to the slide glass with the shiny side facing down. The cells adhered to the top of the membrane were carefully wiped with a Kim wiper or a cotton swab, and then photographed randomly 5 fields per well (200 times magnification) to measure the number of cells. The reaction inhibition rate was calculated according to the following Equation 2 , and the results are shown in Table 3 .
수학식 2
표 3에서 보는 바와 같이, 본 발명의 실시예 1-7의 화합물은 SPC에 의한 세포의 주화성 이동 반응을 강력하게 억제함을 알 수 있었다. 이는 생체 내에서 내피 세포 또는 면역세포의 이동을 억제함으로써, 종양 내부의 혈관 신생, 외부에서 침입한 항원에 대한 면역반응 증폭 등의 과정을 조절할 수 있음을 시사한다. As shown in Table 3, it was found that the compounds of Examples 1-7 of the present invention strongly inhibit the chemotactic migration of cells by SPC. This suggests that by inhibiting the migration of endothelial cells or immune cells in vivo, it is possible to regulate processes such as angiogenesis inside tumors and amplification of immune responses against externally invading antigens.
<실험예 3> 마우스 TPA 유발성 귀 염증 모델에서의 염증 반응 조절실험Experimental Example 3 Inflammation Response Control Experiment in Mouse TPA-Induced Ear Inflammation Model
염증 반응의 억제 효능을 확인하기 위하여, 테트라데카노일 포볼 아세테이트 (Tetradecanoyl phorbol acetate; 이하 TPA) 유발성 염증 모델을 사용하여 다음과 같은 실험을 수행하였다. In order to confirm the inhibitory effect of the inflammatory response, the following experiment was performed using a Tetradecanoyl phorbol acetate (TPA) -induced inflammation model.
TPA 유발성 염증 모델은 염증 반응의 작용기전과 억제물질의 효능을 시험하기 위해 널리 사용되어 온 실험방법이다[De Young LM 등, Agents and Actions, 1989, 26, 335-341]. TPA는 염증 반응을 일으키는 물질로서 피실험체의 귀에 도포하게 되면 홍반과 부종이 발생하게 되는데, 이러한 염증 반응은 백혈구의 세균 공격에 필수 물질인 MPO(myeloperoxidase)의 활성 증가로 측정 가능하다. TPA-induced inflammation models have been widely used to test the mechanism of action of inflammatory responses and the efficacy of inhibitors (De Young LM et al., Agents and Actions , 1989, 26, 335-341). TPA is a substance that causes an inflammatory response, and when applied to the ear of a subject, erythema and edema occur, and this inflammatory response can be measured by increasing activity of MPO (myeloperoxidase), an essential substance for bacterial attack of white blood cells.
6 주령의 ICR 마우스 수컷 40 마리를 준비하여 아세톤에 125 ㎍/㎖ 농도로 녹인 TPA 용액 (시그마 알드리치 코리아)을 왼쪽 귀에 20 ㎕씩 도포하였다. TPA 도포 1시간 후에 아세톤, 아세톤에 녹인 0.3% 실시예의 화합물들, 또는 아세톤에 녹인 0.3% 하이드로코티손(hydrocortisone; 시그마 알드리치 코리아)을 TPA 도포 부위에 20㎕씩 도포하였고, TPA 도포 6시간 후에 동일 부위에 20㎕씩 재도포하였다. TPA 도포 후 24시간이 경과한 시점에 마우스를 경추 탈골법으로 안락사시키고, 왼쪽 귀를 일정한 넓이로 채취하여 무게 및 MPO의 활성을 측정하였다. 반응 억제율은 하기의 수학식 3에 따라 산출하고, 표 4에 나타내었다. 40 male 6-week-old ICR mice were prepared, and 20 μl of TPA solution (Sigma Aldrich Korea) dissolved in acetone at 125 μg / ml was applied to the left ear. After 1 hour of TPA application, 20 μl of acetone, 0.3% Example Compounds dissolved in acetone, or 0.3% hydrocortisone (Sigma Aldrich Korea) dissolved in acetone was applied to the TPA application site, and the same site 6 hours after TPA application. 20 μl was reapplied. At 24 hours after TPA application, the mice were euthanized by cervical dislocation, and the left ear was collected at a constant width to measure weight and MPO activity. Reaction inhibition rate was computed according to following formula (3) , and is shown in Table 4 .
수학식 3
표 4에 나타난 바와 같이, 실시예 1-7 및 1-30의 화합물은 TPA에 의해 유발된 염증 반응을 귀 부종 및 MPO 활성이라는 두 가지 척도 모두에서 유의하게 억제하였으며, 그 효능 강도는 항염증 약물로서 널리 사용되는 하이드로코티손에 대등하였다. 상기 실험결과는 실시예 1-7 및 1-30의 화합물의 피부 항염증 효능이 특히 염증을 일으킨 귀 부위에 호중구가 침윤되는 것을 억제함으로써 이루어졌음을 확인하였다.As shown in Table 4, the compounds of Examples 1-7 and 1-30 significantly inhibited the inflammatory response induced by TPA on both measures of ear edema and MPO activity, and its potency intensity was anti-inflammatory drug. It is comparable to hydrocortisone which is widely used as. The experimental results confirmed that the skin anti-inflammatory efficacy of the compounds of Examples 1-7 and 1-30 was achieved by inhibiting the infiltration of neutrophils into the particularly inflamed ear region.
<제제예 1> 정제(가압 방식)의 제조Preparation Example 1 Preparation of Tablet (Pressure Type)
활성성분으로서, 본 발명의 화학식 1로 표시되는 화합물 5.0 mg을 체로 친 후, 락토오스 14.1 mg, 크로스포비돈 USNF 0.8 mg 및 마그네슘 스테아레이트 0.1 mg을 혼합하고 가압하여 정제로 만들었다.As an active ingredient, 5.0 mg of the compound represented by Chemical Formula 1 of the present invention was sieved, and then tableted by mixing and pressing 14.1 mg of lactose, 0.8 mg of crospovidone USNF and 0.1 mg of magnesium stearate.
<제제예 2> 정제(습식 조립)의 제조Preparation Example 2 Preparation of Tablet (Wet Granulation)
활성성분으로서, 본 발명의 화학식 1로 표시되는 화합물 5.0 mg을 체로 친 후, 락토오스 16.0mg과 녹말 4.0mg을 섞었다. 0.3㎎의 폴리솔베이트80을 순수한 물에 녹인 후 이 용액의 적당량을 첨가한 다음, 미립화하였다. 건조 후에 미립을 체질한 후 콜로이달 실리콘디옥사이드 2.7mg 및 마그네슘 스테아레이트 2.0mg과 섞었다. 미립을 가압하여 정제로 만들었다.As an active ingredient, 5.0 mg of the compound represented by the formula (1) of the present invention was sieved, followed by mixing 16.0 mg of lactose and 4.0 mg of starch. 0.3 mg of Polysorbate 80 was dissolved in pure water and then an appropriate amount of this solution was added and then granulated. After drying, the fine particles were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The granules were pressed into tablets.
<제제예 3> 분말 및 캡슐제의 제조Preparation Example 3 Preparation of Powder and Capsule
활성성분으로서, 본 발명의 화학식 1로 표시되는 화합물 5.0mg을 체로 친 후에, 락토오스 14.8mg, 폴리비닐 피롤리돈 10.0mg, 마그네슘 스테아레이트 0.2mg와 함께 섞었다. 혼합물을 적당한 장치를 사용하여 단단한 No. 5 젤라틴 캡슐에 채웠다. As an active ingredient, 5.0 mg of the compound represented by Formula 1 of the present invention was sieved, and then mixed together with 14.8 mg of lactose, 10.0 mg of polyvinyl pyrrolidone, and 0.2 mg of magnesium stearate. No. solid the mixture using a suitable device. Filled in 5 gelatin capsules.
<제제예 4> 주사제의 제조Preparation Example 4 Preparation of Injection
활성성분으로서, 본 발명의 화학식 1로 표시되는 화합물 100mg을 함유시키고, 그밖에도 만니톨 180mg, Na2HPO4ㆍ12H2O 26mg 및 증류수 2,974mg를 함유시켜 주사제를 제조하였다.As an active ingredient, 100 mg of the compound represented by Formula 1 of the present invention was contained, and in addition, 180 mg of mannitol, 26 mg of Na 2 HPO 4 .12H 2 O, and 2,974 mg of distilled water were used to prepare an injection.
상기에서 살펴본 바와 같이, 본 발명은 As described above, the present invention
첫째, 고체상 조합화학 합성기술을 이용하여 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체, 약제학적으로 허용 가능한 그의 염 또는 통상의 방법으로 분리된 그 광학이성질체를 제공하였고, First, a 2-piperazino-4,5-disubstituted-1,3-thiazole derivative, a pharmaceutically acceptable salt thereof, or an optical isomer thereof separated by conventional methods using solid phase combinatorial chemical synthesis techniques And
둘째, 상기 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체가 사람으로부터 유래된 진피세포 및 마우스를 이용한 동물실험을 통하여 항염증 효과를 검색하여 SPC 수용체에 대한 우수한 억제활성을 규명하였고,Second, the 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives were tested for anti-inflammatory effects through animal experiments using dermal cells and mice derived from humans, and thus excellent inhibition of SPC receptors. Activity was identified,
셋째, 상기 2-피페라지노-4,5-이중치환-1,3-티아졸 유도체 또는 그로부터 분리된 광학이성질체를 유효성분으로 함유하는 SPC 수용체 활성으로 유발되는 염증관련 질환에 유용한 그의 용도를 제공하였다.Third, the use of the 2-piperazino-4,5-disubstituted-1,3-thiazole derivative or an optical isomer isolated therefrom as an active ingredient, which is useful for inflammation-related diseases caused by SPC receptor activity. It was.
이상에서 본 발명은 기재된 실시예에 대해서만 상세히 기술되었지만, 본 발명의 기술사상 범위 내에서 다양한 변형 및 수정이 가능함은 당업자에게 있어서 명백한 것이며, 이러한 변형 및 수정이 첨부된 특허청구범위에 속함은 당연한 것이다.Although the present invention has been described in detail only with respect to the embodiments described, it will be apparent to those skilled in the art that various modifications and variations are possible within the technical spirit of the present invention, and such modifications and modifications belong to the appended claims. .
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| WO2007028999A1 (en) * | 2005-09-09 | 2007-03-15 | Argenta Discovery Limited | Thiazole compounds and their use as pgd2 antagonists |
| WO2007104557A2 (en) * | 2006-03-15 | 2007-09-20 | 4Sc Ag | Novel heterocyclic nf-kb inhibitors |
| WO2007146066A2 (en) * | 2006-06-06 | 2007-12-21 | Critical Therapeutics, Inc. | Novel piperazines, pharmaceutical compositions and methods of use thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20100064952A (en) | 2010-06-15 |
| KR101051077B1 (en) | 2011-07-21 |
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