Disclosure of Invention
The invention aims to design and synthesize a series of novel halogenated pyrimidine compounds containing substituted aniline structures. In vitro activity screening shows that the compounds have excellent inhibitory activity on lung cancer cells, lymphoma cells and leukemia cells, and are expected to be developed into antitumor drugs.
The invention provides a halogenated pyrimidine compound containing a substituted aniline structure, which has a structure shown as a formula (I),
Wherein,
R1 is selected from hydrogen,
X is selected from hydrogen or halogen;
r2 is selected from hydrogen, a 5-to 10-membered aryl or heteroaryl group,The heteroaryl group contains 1 to 3 heteroatoms selected from N, O or S. The aryl or aromatic heterocyclic group is substituted with 1 to 5 identical or different R4;
l is selected from
R3 is selected from 4-10 membered aliphatic cycloalkyl, aliphatic heterocyclic group, aryl or aromatic heterocyclic group, and is simultaneously substituted by 1-5 identical or different R4. The aliphatic heterocyclic group or heteroaryl group contains 1-3 heteroatoms selected from N, O or S.
R4 is selected from hydrogen, hydroxy, halogen, nitro, amino, cyano, morpholinyl, trifluoromethyl, C1-C6 alkyl, C4-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkyl substituted with hydroxy or amino or halogenated, C1-C6 alkylamido, ester, C1-C6 alkylsulfinyl, C1-C6 alkylsulfonyl, C1-C6 alkanoyl, carbamoyl, or carbamoyl substituted with mono or di C1-C6 alkyl.
Further, the halogenated pyrimidine compound containing the substituted aniline structure,
R1 is selected from hydrogen,
X is selected from hydrogen, fluorine, chlorine, bromine and iodine;
R2 is selected from hydrogen, 5-6 membered aryl or heteroaryl,Wherein the heteroaryl contains 1-3 heteroatoms selected from N, O or S and is optionally substituted with 1-3 identical or different R4;
l is selected from
R3 is selected from 4-6 membered aliphatic cycloalkyl, aryl or aromatic heterocycle, simultaneously substituted by 1-3 identical or different R4. The heteroaryl group contains 1 to 3 heteroatoms selected from N, O or S.
R4 is selected from hydrogen, hydroxy, halogen, nitro, amino, cyano, morpholino, trifluoromethyl, C1-C3 alkyl, C1-C6 alkoxy, C1-C6 alkyl substituted with hydroxy or amino or halogenated, C1-C6 alkylamido, ester, carbamoyl.
Furthermore, the halogenated pyrimidine compound medicine containing the substituted aniline structure,
R1 is selected from
X is selected from fluorine;
R2 is selected from hydrogen, 5-6 membered aryl or heteroaryl,Wherein the heteroaryl contains 1-2 heteroatoms selected from N or S, and is optionally substituted with 1-2 identical or different R4;
l is selected from
R3 is selected from 4-6 membered aliphatic cycloalkyl, aryl or aromatic heterocycle, simultaneously substituted by 1-2 identical or different R4. The heteroaryl group contains 1-2N atoms.
R4 is selected from hydrogen, hydroxy, halogen, amino, morpholino, methyl, trifluoromethyl, methoxy.
Further, the halogenated pyrimidine compound containing the substituted aniline structure has the following structural formula:
Compound 22 compound 23 compound 24
A pharmaceutical composition comprises the substituted aniline structure-containing halogenated pyrimidine compound and pharmaceutically acceptable salts thereof as active ingredients and pharmaceutically acceptable excipients.
Preferably, the above pharmaceutical composition, the pharmaceutically acceptable carrier is selected from one or more of filler, disintegrant, binder and lubricant.
Preferably, the pharmaceutical composition is in the form of a tablet, capsule, granule, spray or injection.
The use of a substituted aniline structure-containing halogenated pyrimidine compound or a pharmaceutical composition according to any one of the above in the preparation of a medicament for treating and/or preventing a proliferative disease.
The use of any one of the above substituted aniline structure-containing halogenated pyrimidine compounds or any one of the above pharmaceutical compositions for the preparation of a medicament for the treatment and/or prevention of cancer.
The application of the halogenated pyrimidine compound containing a substituted aniline structure or the pharmaceutical composition in preparing medicines for treating and/or preventing lung cancer, lymphoma and leukemia.
According to some general methods in the art, the substituted aniline structure-containing halogenated pyrimidine compounds shown in the general formula (I) can form pharmaceutically acceptable salts with acids, and pharmaceutically acceptable addition salts include inorganic acid and organic acid addition salts, and salts added with hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid and benzoic acid are particularly preferred.
In addition, prodrugs of the compounds of the present invention are also encompassed by the present invention. Prodrugs of the compounds of the present invention are substituted aniline structure containing halopyrimidines of general formula (I) which may themselves have little to no activity but are converted to the corresponding biologically active form under physiological conditions (e.g., by metabolism, solvolysis or otherwise) after administration.
The beneficial effects of the invention are as follows:
The halogenated pyrimidine compound containing the substituted aniline structure and the pharmaceutically acceptable salt thereof have excellent anti-tumor activity and safety. The in vitro experiments of inhibiting human non-small cell lung cancer cells H1975, human lymphocytic cancer cells REC-1 and human chronic myelogenous leukemia cells K562 prove that the compound has remarkable inhibiting effect on human lung cancer cells, human lymphocytic cancer cells and human leukemia cells, and is a chemical raw material drug, in particular to the preparation of drugs for treating and/or preventing lung cancer, lymphocytic cancer and leukemia.
Detailed Description
The examples and preparations provided below further illustrate and exemplify the compounds of the invention and methods of preparing the same. It should be understood that the scope of the following examples and preparations is not intended to limit the scope of the present invention in any way.
The structure of the halogenated pyrimidine compound containing the substituted aniline structure is shown as a formula (I),
Wherein,
R1 is selected from hydrogen,
X is selected from hydrogen or halogen;
r2 is selected from hydrogen, a 5-to 10-membered aryl or heteroaryl group,The heteroaryl contains 1-3 heteroatoms selected from N, O or S, the aryl or aromatic heterocyclic group is substituted by 1-5 identical or different R4;
l is selected from
R3 is selected from 4-10 membered aliphatic cycloalkyl, aliphatic heterocyclyl, aryl or aromatic heterocyclyl, simultaneously substituted with 1-5 identical or different R4, said aliphatic heterocyclyl or heteroaryl containing 1-3 heteroatoms selected from N, O or S;
R4 is selected from hydrogen, hydroxy, halogen, nitro, amino, cyano, morpholinyl, trifluoromethyl, C1-C6 alkyl, C4-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkyl substituted with hydroxy or amino or halogenated, C1-C6 alkylamido, ester, C1-C6 alkylsulfinyl, C1-C6 alkylsulfonyl, C1-C6 alkanoyl, carbamoyl, or carbamoyl substituted with mono or di C1-C6 alkyl.
The halogenated pyrimidine compound containing the substituted aniline structure,
R1 is selected from hydrogen,
X is selected from hydrogen, fluorine, chlorine, bromine and iodine;
R2 is selected from hydrogen, 5-6 membered aryl or heteroaryl,Wherein the heteroaryl contains 1-3 heteroatoms selected from N, O or S and is optionally substituted with 1-3 identical or different R4;
l is selected from
R3 is selected from 4-6 membered aliphatic cycloalkyl, aryl or aromatic heterocyclic ring, simultaneously substituted by 1-3 identical or different R4, said heteroaryl containing 1-3 heteroatoms selected from N, O or S;
R4 is selected from hydrogen, hydroxy, halogen, nitro, amino, cyano, morpholino, trifluoromethyl, C1-C3 alkyl, C1-C6 alkoxy, C1-C6 alkyl substituted with hydroxy or amino or halogenated, C1-C6 alkylamido, ester, carbamoyl.
The halogenated pyrimidine compound containing the substituted aniline structure,
R1 is selected from
X is selected from fluorine;
R2 is selected from hydrogen, 5-6 membered aryl or heteroaryl,Wherein the heteroaryl contains 1-2 heteroatoms selected from N or S, and is optionally substituted with 1-2 identical or different R4;
l is selected from
R3 is selected from 4-6 membered aliphatic cycloalkyl, aryl or aromatic heterocyclic ring, simultaneously substituted by 1-2 identical or different R4, wherein the heteroaryl contains 1-2N atoms;
R4 is selected from hydrogen, hydroxy, halogen, amino, morpholino, methyl, trifluoromethyl, methoxy.
The halogenated pyrimidine compound containing the substituted aniline structure has the following structural formula:
Compound 4 Compound 5 Compound 6
Compound 37 Compound 38 Compound 39
A pharmaceutical composition is prepared by combining the halogenated pyrimidine compound containing the substituted aniline structure and pharmaceutically acceptable salts thereof as active ingredients with a pharmaceutically acceptable carrier.
The above pharmaceutical composition, wherein the pharmaceutically acceptable carrier is selected from one or more of filler, disintegrant, binder and lubricant.
The medicinal composition is in the form of tablets, capsules, granules, spray or injection.
The application of any one of the halogenated pyrimidine compounds containing substituted aniline structures or any one of the medicinal compositions in preparing medicaments for treating and/or preventing proliferative diseases.
The application of any one of the halogenated pyrimidine compounds containing substituted aniline structures or any one of the medicinal compositions in preparing medicaments for treating and/or preventing cancers.
The tumor is lung cancer, lymphoma, leukemia.
The following synthetic route describes the preparation of substituted aniline structure containing halopyrimidines of general formula (I) according to the invention.
All starting materials are prepared by the manner described in the synthetic schemes below, by methods well known to those of ordinary skill in the art of organic chemistry, or are commercially available. All of the final compounds of the present invention are prepared by the methods described in the synthetic schemes below or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All the variable factors applied in the synthetic schemes one and two below are as defined below or as defined above.
General synthetic route one:
Reagents and conditions:(a)Nitroaniline,DIEA,i-PrOH;(b)R1H,KI,K2CO;(c)Fe-NH4Cl,HOAc,EtOH-H2O;(d)TFA,i-PrOH,N2;(e)Fe-NH4Cl,MeOH-H2O;(f)Acids,HATU,DIEA,DMF;(g)Aldehyde,acetic acid,ethanol;(h)Aldehyde,acetic acid,ethanol,NaBH4.
General synthetic route two:
Reagents and conditions:(a)Iodoaniline,DIEA,i-PrOH;(b)R1H,KI,K2CO3;(c)Fe-NH4Cl,HOAc,EtOH-H2O;(d)TFA,i-PrOH,N2;(e)Boracic acids,Cs2CO3,Pd(PPh3)Cl2,Dioxane;(f)Alkynes,CuI,DIPEA,Dioxane.
The examples are intended to illustrate, but not limit the scope of the invention. The nuclear magnetic resonance hydrogen spectrum of the compound is measured by Bruker ARX-600, and the mass spectrum is measured by Agilent 1100LC/MSD, and all reagents are analytically pure or chemically pure.
The synthetic route of the compounds 1-21 of the general formula (I) is as follows:
Reagents and conditions:(a)3-nitroaniline,DIEA,i-PrOH;(b)Ethylene glycol methyl ether,KI,K2CO3;(c)Fe-NH4Cl,HOAc,EtOH-H2O;(d)TFA,i-PrOH,N2;(e)Fe-NH4Cl,MeOH-H2O;(f)Acids,HATU,DIEA,DMF;(g)Aldehyde,acetic acid,ethanol;(h)Aldehyde,acetic acid,ethanol,NaBH4.
The preparation method comprises the following steps:
Step A preparation of intermediate A2
2, 4-Dichloropyrimidine (5.00 g), m-nitroaniline (4.20 g), and DIEA (5.80 g) were added sequentially to a reaction flask, isopropanol (20 mL) was added to dissolve, and the mixture was transferred to a 90℃oil bath to be reacted under stirring. The reaction was completed by TLC for 12 h. The reaction solution is placed in ice water for cooling, yellow solid is separated out, the filter cake is washed by isopropanol in vacuum filtration, a yellow solid product is obtained, the yellow solid product is transferred to a 45 ℃ oven for vacuum drying, 6.03g of pure yellow solid product intermediate A2 is obtained, and the solid can be directly used for the next reaction.
Step B preparation of intermediate A4
P-fluoronitrobenzene (5.00 g), KI (5.76 g) and K2CO3 (14.39 g) were added sequentially to the flask, dissolved in ethylene glycol methyl ether (20 mL) and transferred to a 110℃oil bath with stirring. The reaction was completed by TLC for 12 h. The reaction solution was cooled to room temperature, the reaction solution was extracted three times with ethyl acetate, saturated NaCl was washed once, dehydrated and dried over anhydrous sodium sulfate, and dried under reduced pressure, to give 6.69g of intermediate A4 as a white solid product.
Step C preparation of intermediate A5
The intermediate A4 (4.00 g) prepared in the step B is placed in a 50mL round bottom flask, absolute ethyl alcohol (27 mL) is added to dissolve the intermediate A4, deionized water (3 mL) and glacial acetic acid (5.00 mL) are added, the mixture is transferred to an 80 ℃ oil bath for stirring reaction, and reduced iron powder (7.95 g) is added in batches. The reaction was allowed to proceed for 4h, and tlc was complete. The reaction solution is filtered by diatomite to remove iron powder while the reaction solution is hot, filtrate is taken, saturated NaHCO3 solution is added to adjust the pH value of the reaction solution to 9-10, diatomite is added again to filter, and emulsification and floccule are removed. The reaction solution was extracted three times with methylene chloride, washed once with saturated NaCl, dehydrated and dried over anhydrous sodium sulfate, and spun-dried under reduced pressure to obtain 3.00g of solid product intermediate A5.
Step D, preparation of intermediate A6
Intermediate A2 (6.00 g) prepared in step A and intermediate A5 (4.75 g) prepared in step 3 were placed in a three-necked flask, isopropanol (20 mL) was added to dissolve, then trifluoroacetic acid (3.82 g) was added, the mixture was evacuated and nitrogen-filled 3 times, and the mixture was transferred to a 90 ℃ oil bath pot to be stirred for reaction. The reaction solution was cooled to room temperature, saturated Na2CO3 solution was added to adjust the pH of the reaction solution to 9-10, the filter cake was rinsed with water to give a yellow solid product, which was transferred to a 45 ℃ oven for vacuum drying to give 6.43g of yellow solid product intermediate A6.
Step E preparation of intermediate A7
Intermediate A6 (2.00 g) and NH4 Cl (0.54 g) prepared in step D were taken and placed in a round bottom flask, and added with absolute methanol (10 mL) to dissolve, deionized water (10 mL) was added, and the mixture was transferred to an 80 ℃ oil bath to be stirred and reacted, after which reduced iron powder (1.13 g) was added in portions. The reaction was carried out for 6h and tlc was complete. The reaction solution is filtered by diatomite to remove iron powder while the reaction solution is hot, filtrate is taken, diatomite is added again for filtering, and emulsification and floccule are removed. The reaction solution is extracted three times by ethyl acetate, saturated NaCl is washed once, anhydrous sodium sulfate is dehydrated and dried, and the crude product is obtained after decompression and spin drying. Purification by column chromatography on silica gel gave 1.67g of product A7 as a white solid.
The preparation method of examples 1 to 16 comprises:
200mg of intermediate A7 (1.0 eq), acid (1.3 eq) and HATU (1.2 eq) were added to a 25mL round bottom flask, dissolved in DMF and stirred at room temperature before triethylamine (3 eq) was added dropwise. The reaction was carried out for 2h and tlc was complete. And adding saturated Na2CO3 aqueous solution into the reaction solution, regulating the pH to 9-10, precipitating white in the reaction solution, adding DCM for extraction three times, washing with saturated saline water once, drying with anhydrous sodium sulfate for removing water, drying under reduced pressure, obtaining a crude product, and separating by column chromatography to obtain the compounds 1-16.
Examples 1 to 16 (see Table I) were each prepared according to the above-mentioned preparation method. Table one:
the preparation method of examples 17 to 18 comprises the following steps:
200mg of key intermediate A7 (1.0 eq) and aldehyde (1.2 eq) were used as raw materials, placed in a 25mL round bottom flask, dissolved in ethanol (10 mL), placed in a 60 ℃ oil bath, stirred, and 5 drops of glacial acetic acid were added dropwise. The reaction is carried out for about 3 hours, and the TCL detection reaction is finished. And adding saturated sodium carbonate aqueous solution to adjust the pH of the reaction solution to 10, precipitating yellow solid in the reaction solution, and carrying out suction filtration by using a funnel to wash the filter cake for multiple times to obtain a yellow solid product. The obtained crude product is pulped by MeOH to obtain pure yellow solid compounds 17-18.
Examples 17 to 18 (see Table II) were each prepared according to the above-mentioned preparation method.
And (II) table:
the preparation method of examples 19 to 21 comprises:
200mg of key intermediate A7 (1.0 eq) and aldehyde (1.2 eq) were used as raw materials, placed in a 25mL round bottom flask, dissolved in ethanol (10 mL), placed in a 60 ℃ oil bath, stirred, and 5 drops of glacial acetic acid were added dropwise. The reaction is carried out for about 3 hours, and the TCL detection reaction is finished. Ethanol (10 mL) was added again to dissolve the mixture after spin-drying, naBH4 (4 eq) was added to the ice bath at 0 ℃ and the mixture was stirred at room temperature to react. The reaction was carried out for 2h and tlc was complete. The reaction solution is extracted three times by methylene dichloride, saturated NaCl is washed once, anhydrous sodium sulfate is dehydrated and dried, and the crude product is obtained by decompression and spin drying. And purifying the obtained solid product through silica gel column chromatography to obtain white solid compounds 19-21.
The compounds of examples 19-21 (see Table III) were prepared separately according to the preparation procedure described above.
Table three:
the synthetic route of the compounds 22-40 of the general formula (I) is as follows:
Reagents and conditions:(a)Iodoaniline,DIEA,i-PrOH;(b)A5,TFA,i-PrOH,N2;(c)Boracic acids,Cs2CO3,Pd(PPh3)Cl2,Dioxane;(d)Alkynes,CuI,DIPEA,Dioxane.
Step F preparation of intermediate A8
2, 4-Dichloro-5-fluoropyrimidine (3.00 g), metaiodoaniline (3.71 g) was placed in a 250mL round bottom flask, DIEA (4.65 g), isopropanol (10 mL) was added, and the mixture was placed in a 90℃oil bath to stir the reaction. After about 6.0 hours of reaction, the TCL test reaction was completed. The reaction solution was cooled to room temperature and then placed in ice water for cooling, a yellow solid was precipitated, after cooling about 0.5h, the mixture was extracted three times with DCM, the organic phases were combined, washed once with saturated NaCl solution, dried over anhydrous Na2SO4, the organic phase was distilled off under reduced pressure, a white solid was precipitated, and 5.02g of pure white solid product intermediate A8 was obtained by suction filtration, which was directly used in the next reaction.
Step G preparation of intermediate A9
Intermediate A8 (6.00 g) prepared in step F and intermediate A5 (4.75 g) prepared in step 3 were placed in a three-necked flask, isopropanol (10 mL) was added to dissolve, then trifluoroacetic acid (3.82 g) was added, the mixture was evacuated and nitrogen-filled 3 times, and the mixture was transferred to a 90 ℃ oil bath pot to be stirred for reaction. The reaction solution is cooled to room temperature, saturated Na2CO3 solution is added to adjust the pH of the reaction solution to 9-10, white solid is separated out from the reaction solution, a filter cake is pumped and filtered by a funnel, the obtained filter cake is washed by water, and the obtained filter cake is placed in a vacuum drying oven at 50 ℃ for drying, so that 2.38g of pure white solid product key intermediate A9 is obtained.
The preparation method of examples 22 to 35 comprises:
200mg of key intermediate A9 (1.0 eq) and boric acid (1.2 eq) are taken as raw materials, the raw materials are placed in a 25mL three-necked flask, dioxane (8 mL) and deionized water (2 mL) are used for dissolving the raw materials, then Cs2CO3 (3.0 eq) are added, N2 is bubbled for 0.5h under normal temperature stirring, pd (PPh3)Cl2(0.1eq),N2) is added and placed in a 100 ℃ oil bath kettle for stirring under the protection of the PPh3)Cl2(0.1eq),N2, the reaction is about 3h, TCL detects that the raw materials are reacted, diatomite is added for suction filtration, the three times of filter cakes are washed by the dioxane, the organic phases are combined, the dioxane is dried in a spinning mode, and the obtained crude product is pulped and purified by MeOH to obtain pure white solid compounds 22-35.
The compounds of examples 22-35 (see Table IV) were prepared separately according to the preparation method described above.
Table four:
the preparation method of examples 36 to 40 comprises the following steps:
200mg of intermediate 9 (1.0 eq) was placed in a 25mL three-necked flask, dissolved in dioxane (10 mL), N2 was bubbled for 0.5h, then ferrocene palladium dichloride (0.1 eq) and DIPEA (3.0 eq) were added, and the mixture was stirred in an 80 ℃ oil bath under the protection of N2, alkyne (1.4 eq) was added, the reaction was allowed to react for about 6h, and the TCL detection was completed. Adding diatomite, carrying out suction filtration, washing the filter cake for three times by using dioxane, carrying out spin drying, and purifying the obtained crude product by column chromatography to obtain pure solid compounds 36-40. The compounds of examples 36-40 (see Table five) were prepared separately according to the preparation method described above.
Table five:
example 41 Compounds 1-40 in vitro anti-tumor cell Activity
The halogenated pyrimidine compounds containing substituted aniline structures and having the general formula (I) according to the present invention were screened for MTT inhibitory activity in vitro against H1975 (non-small cell lung cancer cells), REC-1 (lymphoma cells) and K562 (leukemia cells) cell lines.
(1) After resuscitating and passaging 2-3 times for stabilization, the cells were digested from the bottom of the flask with trypsin solution (0.25%). After pouring the cell digests into a centrifuge tube, the culture broth is then added to terminate digestion. Centrifuging the centrifuge tube at 800r/min for 10min, removing the supernatant, adding 5mL of culture solution, blowing and beating uniformly mixed cells, sucking 10 mu L of cell suspension, adding into a cell counting plate for counting, and adjusting the cell concentration to 104 cells/hole. Except that the A1 well was blank, no cells were added, and 100. Mu.L of the cell suspension was added to the 96-well plate. The 96-well plate was placed in an incubator for cultivation for 24 hours.
(2) The sample was dissolved with 50. Mu.L of dimethyl sulfoxide, then an appropriate amount of culture solution was added to dissolve the sample into 2mg/mL of a liquid medicine, and then the sample was diluted to 20,4,0.8,0.16,0.032. Mu.g/mL in a 24-well plate.
(3) The medicated culture solution in the 96-well plate was discarded, the cells were washed twice with Phosphate Buffered Saline (PBS), 100. Mu.L of MTT (tetrazolium) (0.5 mg/mL) was added to each well, and after 4 hours in the incubator, the MTT solution was discarded, and 100. Mu.L of dimethyl sulfoxide was added. And (3) oscillating on a magnetic oscillator to enable the living cells and MTT reaction product formazan to be fully dissolved, and putting the dissolved cells and MTT reaction product formazan into an enzyme-labeled instrument to measure the result. Drug IC50 values were determined by the Bliss method.
The results of the inhibition of the compounds against H1975 (non-small cell lung cancer cells), REC-1 (lymphoma cells) and K562 (leukemia cells) cell lines are shown in Table six. IC50 in Table six is less than or equal to 2. Mu.M, denoted by A, 10. Mu.M is less than or equal to IC50 > 2. Mu.M, denoted by B, and IC50 is more than 10. Mu.M, denoted by C.
Table six:
As shown in the test results in Table six, the halogenated pyrimidine compounds containing substituted aniline structures in the compounds 1-40 have good inhibitory activity on human non-small cell lung cancer cells H1975, human lymphoma cells REC-1 and human chronic myelogenous leukemia cells K562 in vitro. The compound has good development and application prospects of anti-tumor medicaments.
While the preferred embodiments of the present invention have been described in detail, the present invention is not limited to the specific details of the above embodiments, and various equivalent modifications can be made to the technical solutions of the present invention within the scope of the technical concept of the present invention, and various possible combinations will not be described further in order to avoid unnecessary repetition. Any modification, equivalent replacement or improvement made within the technical concept of the present invention is included in the protection scope of the present invention.