TW424090B - Pyrimidines as folate antimetabolites with selective activity against human brain cancer - Google Patents

Abstract

A series of 2-amino-6-alkyl-5-(4-substituted-1-piperazinyl) pyrimidin-4-one were synthesized, which having the structural formula I, where R1 is hydrogen or alkyl of from one to six carbon atoms; R2 and R3 are independently hydrogen or alkyl of from one to six carbon atoms; vinyl or acety lenyl R4 and R5 are independently, (1) hydrogen, halogen, nitro, cyano, trifluoromethyl, hydroxyl, alkyl of from one to six carbon atoms, alkoxyl of from one to six carbon atoms, or alkanoyl of from one to six carbon atoms; (2) -NR6R7, where R6 and R7 are independently hydrogen, alkyl of from one to six carbon atoms, or alkanoyl of from one to six carbon atoms; (3) -COOR8, where R8 is hydrogen, a pharmaceutically acceptable metal cation, a pharmaceutically acceptable amine cation, or alkyl of from one to six carbon atoms; (4) -CONR9R10 or CONHNR9R10, where R9 and R10 are independently hydrogen, alkyl of from one to six carbon atoms, alkyl of from one to six carbon atoms substituted with one or two carboxyl groups, alkyl of from one to six carbon atoms substituted with one or two carboxyl groups and one -OH, -SH, or -NH2 group, alkyl of from one to six carbon atoms substituted with one or two carboalkoxy groups of from one to six carbon atoms, alkyl of from one to six carbon atoms substituted with one or two carboalkoxy groups of from one to six carbon atoms, and one -OH,-SH, or -NH2 group; (5) R11 is hydrogen, or alkyl of from one to six carbon atoms; (6) -SO2R12, where R12 is hydroxyl, alkyl of from one to six carbon atoms, alkoxy of from one to six carbon atoms, or -NR13, R14 where R13 and R14 are independently hydrogen or alkyl of from one to six carbon atoms or benzyl; and the pharmaceutically acceptable salts thereof. Those compounds showed selective cytotoxicity on human CNS tumor cell line with growth inhibition (GI50) ranged from 1.39 x 10<-7> M to less than 10<-8> M.

Description

4 2: 4090 Printed by A7 B7, Shellfish Consumer Cooperatives, Ministry of Economic Affairs, China. 5. Description of the invention (1) 1 Methotrexate is a dihydrofolate reductase inhibitor, long-term It has been used as an antitumor agent since then. However, according to Barakat, RR et al., 1993, Gynecol. Oncol., Vol. 51, p. 5 54-60,-the drug has fatal injuries that are resistant to the drug. Question. The production of this drug resistant branch was studied by Morris, JA et al. In 1994, Biochem. Pharmacol., Volume 47, Nos. 1207-20, due to changes in the enzymes of the article where the drug acts; due to ΙΜχοη, K.Η. et al., 1994, J. Mol. Chem. Vol. 269 vol. 17-20 K. reported that the activity of folate carrrier was reduced in 10 siblings, causing the drug The soft sensory skin of the transport deficient cell that has been damaged is reduced. The inventor has synthesized a series of folate metabolism antagonists belonging to the 15 piperazine-pyrimidines chemical structure, of which 2 ,, 4-diamino-5-hexa-JL Symptoms (2,4_d iam ino_5-pi perazine-pyriniidines), because they can freely penetrate cells to reach the target enzymes to exert the effect of inhibiting enzymes, and have fine toxicities to mouse leukemia (P388 L1210) and sarcoma (sarcoma) ( cytotoxicity) 〇Some of these have relatively high fat-soluble folate metabolism antagonists * reported by Fry, DW, et al., 1986, P. oc.Am. Assoc. Cancer Res. Vol. 27, Vol. 253 K. The penetration of the fine clinging membrane enhances absorption, so the drug has impaired folate transport mechanism and cannot absorb drug resistant cancer cells (such as Ul-L2 / Mr MTX) based on methotrexate, which can exert a good effect of 24 (Please read the precautions on the back before filling this page] Γ The size of the paper method is applicable to China National Standards (CNS) A4 (2 丨 0x297 mm) 4 424090 Employees of the Central Standards Bureau of the Ministry of Economic Affairs Cooperative printed A7 B7 V. Description of Invention (2) 1 year ^ Baldwin SW and Taylor, EC, etc. A ^ V 1991

A series of reports bought in Biochemistry Volume 30, 1997-2006, mentioning a powerful effect of 5,1 on a mammalian Glycinamide Ribonucleotide transformylase (50 servings &quot; ^ 380). -Bis-free. Nitro-5,6,7,8-tetrahydrofolic acid (DDATHF, 5,10 ~ dideaza-5,6,7,8-tetrahydrofο 1 ic ac id). This compound has become a clinical experimental drug-Lometrex Clometrexc 1). According to Benkovic, SJ, Biochem * Sci., Vol. 9, pp. 320-22, 1984, this daramine ribonucleoside methyltransferase (GAR-Tfase) 10 is synthesized in de novo One-carbon-unit transfer reaction ion in Biosynthesis) uses amidinotetra &amp; folic acid (N5, H10-methenyltetrahydrOfolic acid »KTHF) as a cofactor. Therefore, the discovery of the drug Luomentuo belongs to a new pharmacological mechanism. According to Natsumeda, Y. et al., 1984. 15 Cancer Res. Spring 44 No. 2475-79 I, cancer cells usually have a higher rate of de novo Biosynthesis than normal cells. Sexually inhibits the activity of daramine riboside glycosyltransferase (GAR-Tfase) in mammals and has become one of the cytotoxic agents for developing cancer cells. 0 20

Kelly, J. 11 Bigham * EC * Harrington ♦ PM · and Hodson, SJ · et al. In 1990 J. Med. Cheni. Vol. 33 No. 561 671 * 1992 丄 Med. Chein Vol. 35 No. 1399- 410 pages, 1992. J. med. Cheffl Vol. 35, 1109-116K, and 19944 J. med. Chem. Vol. 24, 37, pp. 2112-115 report a series of chain-like tetrahydrofolate forms (please read first (Notes on the back then fill out this page)

Tong Guo 4I4Q90 A7 ------- B7 V. Description of the Invention (3) '′ — 1 thing (Acyclic Analogue of 5,6 · 7, δ-Tetrahydrofolic

Synthesis of acid type compounds and physiological activities. These compounds are chain-shaped pyrimidine ring-opening compounds (〇pen-chain analogues) developed for the difficulty of preparing DDATHF. However, their activities are less than DDATHF. According to the previous literature, It is possible that the degree of freedom of the branched structure of this series of ring-opening compounds is too large to maintain the conformation of methylenetetrahydrofolate. In view of the above, in order to increase lipophilicity, Free penetration and absorption into the cell membrane, as well as maintaining structural elasticity, and conforming to the spatial S &amp; position of fluorenyltetrahydrofolate, the present invention is a series of pyrimidine folate metabolism antagonists with special activity on brain cancer cells. A ring-opening compound with methylenetetrahydrofolate (MTHF>), that is, 2-amino-6-carboxyl-5_ (4-substituted-1-hexahydropyridine) pyrimidine-4- (3H ) -Methyl compounds. This class of 15 compounds approximates the spatial configuration of methylenetetrahydrofolate (spacia 1 or * ie- ntation) »After preliminary anti-cancer screening, it has been shown to be quite active for human brain cells. Central Bureau of Standards, Ministry of Economic Affairs Printed by a consumer cooperative ----- „---- i-- (please read the precautions on the back before filling out this page) ^ _ The present invention has a special vermin-like pyrimidine folate metabolism on brain cancer cells Antagonist &quot; 2-Amine-based total-5- (4-substituted-1-hexahydropyridine) pyrimidine-4- 20 (3H) _fluorenyl structure compound is prepared in a polar organic solvent containing the test The reaction between the formula (I) and the formula (B0 is shown in Figure ㈡ 'Reformation of the shifted class * when necessary to form a pharmaceutically acceptable salt. Wherein Ri of the formula 0 [) is hydrogen or a branched group, and R2 * R3 respectively represent Amino group, school group, and hexyl group are formed. R 4 and 5 represent electron-extracting substituents. Dangerous organic solvents refer to alcohol solutions containing sodium hydroxide 24 and bell hydroxide, usually having less than 6 carbon atoms. ^ 1 This paper is a standard for domestic use (CNS) Α4 (2 丨 0 × 297 公 嫠 A7

[I III Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs 414090 V. Description of the invention (4) Low-grade drunk * The more appropriate low-grade 蜉 颏 is methanol, ethanol, and propionate. The physical property data of the compounds prepared according to the above methods were measured by various instruments, respectively.

H2N 10 The pyramidine-type folate metabolizing agent with special activity on brain cancer cells of the present invention &quot; 2-amino-6-amino-5- (4-substituted-1-hexahydropyrimidine-4- (3H) -Instantaneous structure compounding field is prepared as shown in Figure ，. N-benzylpiperazine is ethyl 2-oxo_3 ~ chlon &gt; propionate) Condensation &gt; 6 ^ ## 15 Compound 3 »Condensation with guanidine to obtain 1 pyrimidine-containing compound 4. Compound 4 in hydrogen and # a- Hydrogenation under the environment of “palladium on charcoal” to produce a debenzylation to obtain compound 5. Compound 5 is compounded with 4-fluorobenzonitrile and acidified with water after 2 Decomposition treatment to make compound 7. Compound 7 was compounded with diethyl glutamate by didi glutamate (EDCI, l ~ ethyl-3- (3-dimethyl arainopropyl carbodiimide)). δ. By adding saponification solution containing sodium argon oxide to saponification reaction, product 9 is obtained. _24__Comparing to the preparation method of compound 9, compound 10-14 can be obtained, in which Zhang scale appropriate national fiscal provision f (CNS) from Peru (χ 297 other male bags) 7 (please pay attention to the back of the Mtt r key and then fill in this page)

Printed by A7 B7 of the Consumers' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 5. Description of the Invention (5) 1 The main difference is only that compounds 10-14 are structurally free of short-term acid, so compounds 5 and 4-fluoronitrobenzene ( 4-fluorOnitrobenzene> is combined to obtain compound 10. Compound 5 is condensed with N, N-dibenzyl 4-fluorObenzenesulfonamide &gt; 5 (Condensation) to obtain compound 22, which Then, the compound 22 is subjected to dehydration to obtain 4 di-compound 12. The hydrazone 6 is reduced by argon hydrogenation to obtain compound 13 and compound 14 is esterified. 10 The pyrimidine-type folate metabolism antagonist of the present invention, which has special activity on brain cancer cells, is a compound of the formula (I) and a pharmaceutically acceptable compound thereof, wherein R 1 is a aryl group or ^, R2 and R3 may respectively represent hydrogen, xyl, vinyl or ethynyl; RfR5 may respectively

15 (1) .Gasyl, Nansu, Nitro, 11 &amp;, trifluorofluorenyl, light radical, CV 6 sulfo, C6 alkyl, (alkoxyl), C6 6 alkanoyl ⑵ -NR6R ", where R6 and R7 may respectively represent a hydrogen group, a ^ _6 alkyl group, C1-6 (a lkanoy 1) 20 (3). ~ C00R8, where 1 to 8 are hydrogen group, Cu school group, pharmaceutically acceptable Jinzhan ion, pharmaceutically acceptable amine ion, (4), ~ 〇0 服 91〇 and ~ 〇) 1 «9 feet 10, of which 1 ^ 9, trans: 10 can represent alkenyl, Cu school, and be C 1-6 substituted by one or two acyl groups, substituted by one or two acyl groups and a single acyl group, SH or amine group—24_ Ci-6 alkyl group, substituted by one or two Ci-6 alkyl groups Replaced Ci-6 This paper size is applicable to China Standards (CNS) A4 specifications (2 丨 0X297 mm (please read the note on the back before filling this page) -s A7 A24090 5. Description of the invention (. 6). 1. C 1-6 alkyl substituted with-or one C 6 alkane s |; g ^ single, S} {or amine group. ⑸ * 卞 ㈣ where R 11 is a hydrogen group, 0: Bu 6 5 -· -S02R12, where R12 is meridian, Cl-6, Cl-6 alkoxy; or -HR13R14, where Ri3 and R14 can be fermented, respectively. 6 silk, benzyl , 0 ‘II, —-^ ------------- '^. (Please read the precautions on the back before filling in this page)

ο 4 2 2 Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs. The present invention is a vocal folic acid metabolism antagonist with special activity on the purchase of S cells. Because of its considerable fat solubility, it may be able to penetrate fine buttons freely. Due to the change in the absorption of the small ship, the carbamate folic acid (methotre- this paper size applies to the Chinese national standard ((: yang) 8 4 kampan (2 丨 0 &gt; &lt; 297)) 9 Central Bureau of Standards, Ministry of Economic Affairs Printed by the Industrial and Commercial Cooperative Society 42 In 2009, the description of the invention (7) 1 xate) Cancer cells that are resistant to absorption and have drug resistance have selective cytotox icity, so they can be used for human and animal brain cancer The above formula (I) can be used to form enamels with organic acids and pharmacologically tolerable acetic acid. Various excipients such as odorants, #release agents (diluents), lubricants, and mixtures can be added. (binders), disintegrations (disinteg-rants), or colorants, agents made into lozenges or other solid preparations; and phosphate phosphate buffer solution to adjust the pH value can be made into injections or other liquids and various dosage forms .Pharmaceuticals of the compound of formula (I) of the present invention Both the acid addition M and the pharmaceutical composition which can tolerate the acid addition can produce 10 tumor cell inhibitory activity. * The general dosage can be adjusted according to the symptoms, usually 20 mg to 300 per person per time. mg, 3 times a day. The compound of the present invention, which has special activity on pyrimidine folic acid metabolism inhibitors &quot; for brain cancer cells, has undergone antitumor with 60 15 kinds of cancer cells by the National Cancer Research Center The results of the screening are shown in Table ㈠. The GI50 value in Table 代表 indicates that the growth equivalent to 50% of the cancer's embracing at that range is inhibited. Most of the compounds of the present invention are effective for cancers in the middle frame system. Cells (CNS cancer subpanel) have selective cytotoxicity, in which compounds 20, 9, 10 and 20 have a GI50 value lower than 10-8 M. Compound 11 is effective against cancer, colorectal cancer, ovary, kidney , Prostate, breast cancer and anti-brain cancer are all active. Preparation Example 1 2Α -__ ethyl 2- (4-fluorenylhexahydro pillow whistle HH) -3 Jinqi Ding (e + hvl t 用 中国 @ 家 县 （ CNS) Α4 · (210X297 read) ~ ----- {Please read the note on the back first Item and then fill in this page)

Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Health 42 ^ 09π Α7 __ _Β7 ___- V. Description of the Invention (8) 1 2 ~ (4-phenylmethylpiperazin -l-yl) -3-oxo- butyrate * 3) Will be 85.15 g ( 0.52 mmol) of 5 ethyl 3-chloro-2-oxopropionate and 6.42 g (0.49 mol (AcCN) of 7.1 g (0.52 mmol)) of potassium carbonate at 50 Stir for 24 hours at ° C. Remove acetcyanide (AcCK) under low pressure, dissolve the residue in 5% 300 mL of sodium carbonate aqueous solution and partition twice with 300 mL of ethyl acetate (EtOAC). 10 Collect the ethyl acetate The ester layer was extracted with 1 N hanging acid, and the aqueous solution was separated. The neutralized solution was neutralized to pH 9 and extracted twice with 250 mL of ethyl acetate. The ethyl acetate layer was separated, dried over sulfuric acid, and filtered. The filtrate was concentrated to obtain 132.60 g of a light yellow liquid product with a yield of δ9%. 15 Preparation Example 2 2-Amine-6 ~ Tyl-5 (4_Mentylhexahydroca-H) ring syndrome ~ 4 ( 3Η) (2-Am i no ~ 6 &quot; methy 1-5- (4-pheny 1 methy 1- piperazine-l-yl) pyrimidin-4 (3H) -〇ne 4) 20 Take 2.43 g (0.11 mmol) ) Sodium is dissolved in 150 mL of 2-nethoxyethanol, Add 30.40 g (0.10 mol) of compound 3 and · 10.03 g (0.11 mmol) of guanidine HC1 (guanidine HC1), the mixture is heated and refluxed for 13 hours. The dissolved coal is removed under a low pressure state, and the residue is Dissolved in 300 mL of water, collected the precipitate, and recrystallized with 95% ethanol to obtain 17.54 g of 24 white solids with a yield of 95%. (Jingxian Mi * Note on the back of the deposit please fill in this note) ^.

) y »J This paper is again applicable to the Chinese National Standard (〇 奶) 六 格 (21〇297297mm) 11 42409〇Α7 B7 V. Description of the invention (9) 1 Preliminary Example 3 2-Amine- 6-fluorenyl ** 5- (4-hexahydrola-1) yrididine-4⑶-yin (2 ~ Am in〇-6-methy 1 -5- (4-pi peraz i ne-1 -y 1) pyrimidin-4 (3H) ^ one · 5) 5 Dissolve 16.56 g (5.50 mmol) of compound 4 in 200 mL of glacial acetic acid solution, add 0.5 g of 10% Pd / C catalyst to hydrogenation under 1 atm hydrogen Reaction (hydrogenolysis), complete reaction over 34 hours. The catalyst was filtered off, the dissolved coal was removed in a low pressure state, recrystallized with hot wood, and dried at δ0 in a low pressure state overnight to obtain 10.67 g of compound 5 as white needle crystals, with a yield of 92%. Example 1 15 4- [4- (2-Amino-6-methyl-4 (3H) -instantyl salivary-5-yl) liuyinggu 11 丼, 1- &amp;] phenylnitrile (4 -[4- (2-Am in〇-6-methy 1-4 (3H) -0X0-pyrimidin-5-yl) piperazin-l-yl] benzonitrile »6) 8.70 g (41.63 mol) mixed Compound 5, 5.10 g (42.15 20 mol) of 4-fluorobenzonitrile was reacted with 6.89 g (49.96 mmol of sodium perforate in 250 mL of DMF) and heated under reflux for 7 hours to complete the reaction. Remove the solvent in the low pressure state, dissolve the residue in 250 mL of water, collect the precipitate, recrystallize it in DMF, and place it in SO'C under low pressure to dry it well. 8.20 g of compound 6 is obtained as white needle crystals, with a yield of 24 It is 64%. ---------- X — (Please read the notes on the back before filling in this page) Order printed by the Central Bureau of Standards of the Ministry of Economic Affairs, Shellfish Consumer Cooperative, printed on paper, suitable for Central European countries Standard Half (CNS) Eighty-four Regulations (2 丨 0 乂 297mm) Α7 42409η V. Description of Invention (1〇) 1 Example 2 II ---- ---- IIII Order {Jing first read Note on the back, please fill in this page again) 4- [4 ~ (2-Amino-6-methyl-4 (3H) -ketoconid-5-yl) hexahydro 11 丼 -1-yl] benzoic acid (4- [4- (2-Afflino “6-methy 1 -4 (3H) -5 〇xopyrimidin-5-yl) piperazin-l-yl] benzonic acid» 7) Dissolve 6.07 ε (19.40 mmol of phosphonium compound 6 in 100 mL of concentrated citric acid and heat under reflux for 12 hours. Remove the solvent in a low pressure state. • Residue dissolved in ice water 10, collect the precipitate, and recrystallize with DMF. It was placed in a low-pressure state at 80 ° C and dried too well to obtain 4.85 g of compound 7 with white needle crystals in a yield of 75%. Example 3 15 Diethyl [4- [4- (2-amino-6- F radical-4 (3H) -fluorenyl yell -5 _yl) hexahydrocai-1-yl] stupid: histone ①iethyl H- [4- [4- (2- Amino -6- methyl -4 (3H)-oxopyrimidin -5 -yl) piperazin-l-yl] benzoyl] glutamic acid 1 8) Printed by the Central Standards Bureau of the Ministry of Economy Compound 7, 2.118 (11.00 mmol), EDCI, 1-ethy, 3- (3-dimethylaminopropyI) car * bodHmide, 2.40 s (10.00 mmol, 6 base ft gas acid cyanic acid H (diethyl glutamate hydrochloride ^ 1.36 g (10.00 24 hydroxybenzotr * iazo 1 e monohydrate, paper size Using China National Slope (CNS) Eight-four Rule Pan (2 丨 OXW7mm) I] 42409ο A7 42409ο A7 Printed by the Bayer Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs Ⅴ. Description of the Invention (11) 1 HOBt), 2.11 g (10.00 mmol of methyl morpholine (· ,. methyl morpholine) was dissolved in 150 niL of DMF, and incubated at 50 t for 28 hours. The solvent was removed in a low pressure state, and the residue was washed successively with ethyl acetate and water, and unreacted reactants were removed by ultrasonic vibration. The precipitate was collected, crystallized with 95% 5 ethanol hydrazone, and dried at 80 ° C under a low pressure condition to obtain a good yield of 4.05 ′ g of Heyang Yang 8, with a yield of 79%. Example 4 10 N- [4- [4- (2-Yenyl-6-methyl-4 (3H> -gangylhanid-5-yl) hexaoxy late β well · * 1_yl] benzyl ammonia Acid (N ~ [4_ [4- (2-amino-6 ~ methy 1 -4 (3H) -〇xopyrimidin-5-yl) piperazin-l-yl] benzoyl] glutamic acid »9) 15 2.16 g (4.20 The mmol δ δ was dissolved in 30 mL of 3 i {hydroxide, stirred for 7 hours, and then treated with 1 N acid, and the precipitate was collected by filtration. * Recrystallized with an aqueous solution containing DHF, and placed in a low pressure state 80 cC was dried over rhenium to obtain 1.63 g of compound 9 with a yield of 85%. 20 Example 5 2-Amino-6 ** methyl-5- [4- (4-acylbenzene) hexahydro 11 and 1-yl) _4 (3H) -one (2 ~ Am ino-6-inethy 1 -5- [4- (4_n i tropheny 1) pipera2ine-l-yl) pyriinidin-4 (3H) -〇ne * 10) This paper size applies to Chinese national standards (€ Yang &gt; Six Shapes 1 Cell (2 丨 0 Father 297 Gongkang 14 (Please read the note on the back-• iW 'before filling this page)]

1 Α7 V. Description of the invention (12) 1 1.00 g (4.780 ^ mole of compound 5, 0. 67 S (4.780 mmol of 4-gasified nitrobenzene (4-1 * 111〇 「〇11 丨 1 ^ 〇1) 01 ^) and 0.666 8 (4.78 mmol) carbonic acid was mixed with 50 mL of ethanol and heated under reflux for 11 hours. The solvent DMF was removed under a low pressure state, and the residue was washed with 50 raL of ethyl acetate. 5 Filtration to remove soluble reactants. The solid residue was dissolved in 50 虬 of water and recrystallized with DMF to obtain 1.21 g of white needle-like crystals with a yield of 77%. Example 6 10 0-Dimethyl 4- [4- (2-Amino-6-methyl-4 (3H) &amp; pyrimidine- 5-yl) hexakiazi-1-yl] phenylacetamine (N, N-pibenzyl —WG-am ino-6-methy 卜 4 (3H) -oxopyr ί in idi n-5-y 1) pi peraz in-1 -yl] benzenesulfonainide &gt; 11) 15 Employees of the Central Bureau of Standards of the Ministry of Economic Affairs * k Cooperative print- II-I---I-I》-^ I-I-1-(Please read the precautions on the back before filling out this page) Put 3.55 9 (110.66 mmol of 2-year-based 4-fluorobenzylamine ( N, N-dibenzyl 4-fluorobenzenesulfonamide) and 2.09 g (20 10.00 mmol) of compound 5, 1.52 8 (11.00 mmol) of carbon Sodium acid is mixed with 25 raL of DMSO * heated under reflux for 9 hours. Remove the solvent DHS0 in a low temperature state, put the residue in 100 虬 ice water to help the formation of precipitate, collect the solid solution and use 2-methoxyethanol ( 2-methoxyethano 丨) was recrystallized and placed under low pressure at 80 t to dry overnight to obtain 3.41 g of white solid 24 with a yield of 63%. This paper size is applicable to China Storehouse Standard (CNS) A4iMM 210X 297 mm Central of Ministry of Economic Affairs Printed by the Bureau of Standardization of Shellfish Consumer Cooperatives 424090 A7 _B7___ V. Description of the Invention (13) 1 Example 7 4- [4- (2-Amino-6-Sub-based-4 (3H) -Hydroxy-based yell bite-5- Hexahydrogen β--1-yl] benzene continued to press ('Μ-^-βηιίηο-β-ιιίθΙ ^ Ι-ΑβίΟ-οχο-δ pyr imidin ~ 5 ~ y 1) p iperazin ~ ly 1] benzenesu 1 fonain ide * 12)

Place 1.36 g (2.500 mmol of Compound 11) in ice-cold 20 mL of Tear Sulfuric Acid, and simmer for 7.5 hours. After the reaction is complete, pour into 50 mL of 10 ice water, and partition with 50 mL of ethyl acetate (EtOAC) The water shoulder was collected to neutralize with 6 N sodium argon oxide, the precipitate was collected, the solid was washed with water and recrystallized with DMF to obtain a 0.38 s white solid with a yield of 91%. Example δ 15 2-amino-5 [4 (4-Aminomethylphenyl) hexahydropyrine-1-yl) 6 ~ methylpyrimidine-4 ~ (3H) ~ Sl ^ (2 — Amino — 5 [4 (4 ~ aminomethylphenyl) piperazine-1- yl) 6 -raethylpyrimidin- 4- (3H) -one »13) 20 * 13) Dissolve 0.10 g (0.320 mmol) of compound 6 in 20 mL of acetone solution containing 0.5 g of acetic acid, and add 0.03 g of 10% Pd / C catalyst, hydrogenolysis under 3.5 atm argon, complete reaction in 50 hours. Filter off the catalyst, remove the solvent in a low pressure state, pour the residue into a diluted aqueous solution of sodium bicarbonate, collect the sink, and recrystallize with 95% ethanol "(Please read the precautions on the back before filling this page)

T ^ Zhang Niu is suitable for the Chinese national ladder car (CNS ... Lin (2'297 public * U6 __ A7 B7) V. Description of the invention (14) 1 Place it at δ 0 ° C under low pressure to dry overnight to obtain δ2.0 g of white powder. The yield is 92%. 5 Example 9 4- [4- (2-Amino-6-methyl-4 (3H) -fluorenyl-5-yl) -hexahydro 0 ratio &quot; Well- 1-yl] benzoic acid ethyl vinegar (4- [4- (2-ainiiio-6 ~ me1; hyl-4 (3H) -oxo-pyrimidin-5-yl) piperazin-l-yl] benzoic acid ethyl ester, 14) 10 15 Mix 0.30 g (0.91 mmol) of compound 7 into 50 mL of 95% ethanol solution containing 1.0% sulfuric acid and heat under reflux for 22 hours. Remove the solvent under low pressure and pour the residue into 100 mL of ice. Water and neutralized with sodium diacetate, the precipitate was collected and washed with 50 ni of water, recrystallized with 95% ethanol, and dried at 80 ° C under low pressure overnight to obtain 4.05 g of white powder. The yield was 79¾. Example 10 I ί ·---I II tn (Please read the note f on the back of the book before filling in this page) Printed by Zhengong Consumer Cooperative of Central Bureau of Samples of the Ministry of Economic Affairs 20 24 Main ingredients 50 nig Lactose 30 mg Starch 4 mg magnesium stearate 6 mg corn flour 10 mg

ML The paper scale is applicable to Chinese family standards (CNS &gt; People 4 Rules Brigade (210X 297 mm) 17 A7 JV \ .j B7 V. Description of the invention (15) 1 According to the above formula, the main product price can be obtained Illustrative drawing of cocoons in lozenges Figure 1 ... Aminofolate 2 ... Methylenetetrahydrofolate 5 3 ... 5,10-Didezazide-5,6,7, δ-Tetra Hydrofolate 4.,. The target compound Figure 2 Reaction scheme 扃 Compound 3 preparation method Table 1 Antitumor activity 10 Table 2 Physical data (Please read the precautions on the back before filling this page)

* 1T Consumption cooperation for employees of the Central Prototype Bureau of the Ministry of Economic Affairs. Printed on China National Standard Vehicle (CNS) Α4 (210X297 mm) 424G9 A7 B7 V. Description of Invention (16) ^ 丨 a250 ί 50% s许 洳 Ai s 6 7 CC910 11 12 -cz: h!, Co§!! -CO2H- s- (nE2) 2COOEt no〇Et ί! CONH--oH- (CH2) 2COOH no〇a!: -2: 02!-, So2N (CH2ph) 2 4.8-79 55 _S02NH2_ I: (Please read the precautions on the back before filling out this page) Employees' Cooperatives of the Central Standards Bureau of the Ministry of Economics Seal 0.04 5 Λ ° .ο 1 丨 · 丨 丨-丨 <0.01 —I __ i II —Λ0.01 — in! • 68 p'-3r27 1.9-3.5 Zr37 0 '1 ° I — — — 1.39 gallium 齑 谇 谇 〇150, port 2) „

1T This paper size is applicable to the China Solid State Mou (0: Mi) 8 4 specifications (21 〇 father 297 mm &gt; 19 printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 4240SQ A7 B7 V. Description of the invention (port)

Table II a W

Ethyl 2- (4-phenylmethy) piperazin-l-yl) -3- &lt;) x〇-butyrate (3) IR (KBr): 3011, 2940, 2810, 1740, 1720, 1650, 1358, 1325, 1250, 1016 cm-1; 1H NMR (80 Mz, CDC13): δ 1.17-1.24 (t, J = 7.1 Hz, 3H, C ^ CHgOCO-), 2.05 (s, 3H, CH3CO-), 2.40-2.50 (π, 4H, piperazinyJ-H), 2.60-2.70 (id, 4H, piperazinyl-H), 3.45 (s, 2H, -CH2-Ph), 3.84 (s, 1H, CH3CO-CH-), 4.11-4.18 (qT J = 7.1 Hz, 2H, CH3CH2OCO-), 7.10-7.29 (m, 5H, Ph-H) ppm;

El, MS a / z (relative iiitensity); 304 CM +, 13), 261 (100), 231 (10), 170 (29), 91 (95). 2-Amino-6-methyI-5- (4- phenyImethylpiperazin-l-yI) pyrimidin-4 (3H) -one (4) nip: 280-281 ° C; IR (KBr): 3350 (br), 3160, 3080, 2940, 2850, 1650, 1610, 1540, 1390 cra ~ l; 1H NHR (80 MHz, DMS0-d6): &lt; 5 2.07 (s, 3H, CH3-), 2.25-2.65 (id, 4H, piperazinyl-H), 2.75-3.10 (id, 4H, piperazinyl -H), 5.9-6.4 (brs, 2H, -NH2), 7.Π-7.4 (m, 5H. Ph-H) ddbi; ---------- Wood— (Please read the Please fill in this page for the matters needing attention) The paper rule A of this edition uses the Chinese National Standard (CNS) 210X297 Public Love &gt; 2〇 Printing by the Consumers' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 414090 A7 B7 V. Description of the invention (18) , Table 2b

El MS b / z ^ relative intensity): 299 30), 208 (20), 180 (10), 165 (20), 153 (53), 146 (19), 91 (100);

Anal for C16H21N50 · 1/2 H2〇: Calcd. C, 62.32; H, 7.19; N, 22.71;

Found C, 6L99; H, 7.34; N, 22.75, 2-Ami no-6-methyl-5- (4-piperazir) ~ l-yl) pyriiDidin-4 (3H) -one (5) rap: 285-286 ° C; IR (KBr): 3350 (br), 3180, 2950, 2850, 1655, 1350, 1240 cm-J; NMR (80 MHz, D2〇 + NaOH): ¢ 5 2.59 (s, 3H, CH3-) ; 3.05-3.50 (jh, 8H, piperazinyl-H) ppm;

El MS si / z (^ relative intensity); 209 (M +, 45), 167 (100), 153 (23);

Anal for C9H15N50: Calcd. C, 49.53; H, 7.39; N, 32.09;

Found C, 49.39; H, 7.55; N, 32.03. 4- (4- (2-Am i no-6-methy1-4 (3fl) -oxopyr imidi n-5-y1) pi peraz in-1-yl) benzonitriie (6) rap: 330 ° C (deep); IR (KBr): 3125, 2900, 2860, 2220, 1680, 1620, 1580, 1515, 1440, 1380, 1320, 1275, 1240, 1180, 1100, 1040 caj -I; This paper size applies to the Chinese family rate (CMS) A4 secret (210X297 mm &gt; 21 (Jing first read the note on the back and fill in this page)% Order

4 2409Q A7 B7 V. Description of the invention (19) Table 2. 1H NMR (80 HHz, MSO-d6): 5 2.13 Cs, 3H, Cfl3-), 2.90-3.20 (id, 4H, piperazinyl-H), 3.21-3.50 (id, 4fl, piperazinyl-H), 6.12-6.45 (brs, 2H, -NH2), 6.85-7.15 &amp; 7.40-7.70 (ABq, J = 8.8 Hz, 4H, Ph-H) ppm;

El MS s / z (relative intensity): 310 CM +, 100), 295 (48). 179 C15), 166 (13), 153 (97), 129 (30), 102 (25), 73 (20); HRHS Calcd. For CigHigNgO: 310.1542; Found 310.1545. 4- [4- (2-Amino-6-methyl-4 (3H) -〇x〇pyrin] idin-5-yl) piperazin-ly]] benzoic acid (7 ) rap: 295 ^ 296 ° C (deep); IR (KBr): 3400 Cbr), 3150, 2975, 2850, 1680, 1650. 1610, 1520, 1380, 1240, 1190 cm-1; 1H NMR (80 MHz, DHSO-d6): 6 1.15-1.45 (t, J = 7.04 Hz, 3H, CH3CH2-), 2.12 Cs, 3H, -CH3) 2.8-3.5 (m, 8H, piperazinyl-H), 4.05-4.45 (q, J = 7.04 Hz, 2H, CH3q ^), 6.15-6.45 (br, 2H, -¾). '6.80-7.15 &amp; 7.40-7.70 (ABq, J = 8.92 Hz, 4H, Ph-H) ppra; Ministry of Economic Affairs Printed bags of the Central Bureau of Standards, Shellfish Consumer Cooperatives (please read the note on the back before filling this page)

El MS tn / z (relative intensity): 357 OiK 35), 342 (35), 312 (11), 179 (21), 166 (20), 153 (100), this paper scale is applicable to China's national ladder (CNS ) Α4 specifications (210 × 297 mm A7 42409 ^ B7 V. Description of the invention (20) Table d 138 (10), 132 (25), 125 (21). HEMS Calcd. For C16H19N503: 329.1487; Found 329.1492. (Please first (M read the note on the back and fill in this page)

Diethyl N-C4- (4 ~ (2 &quot; Aniino-6_methy) -4 (3H) -oxopyrinidin-5_y 1) piperazin ~ ly]) benzoy] JgJutamic acid (8) rap: 290-291 ° C; IR CKBr): 3370 (br), 3150, 2980, 2850, 1738, 1.730, 1660. 1640, 1610, 1510, 1230 cra'l; 1H-NMR (400 MHz, DHS0-d6); 5 1.1-1.2 (id, 6H, 2CH3CH2 -), 1.9- 2.0 (π, 2H, -CH2CH2COOEt), 2.1 (s, 3H, CH3), 2.4 (id, 2H. -CH2C00Et), 2.9- 3.5 (id, 8H, piperazinyl-H), 4.0-4.1 («Η. 4H · 2CH3Cii2-), 4.4-4.5 (m, 1H, -CONHCH), 6.3 Cs, 2H, NH2), 6.9 &amp; 7.7 (ABq, J = 8.8 Hz, 4H, Ph-H), 8.3 (d, J = 7.6 Hz, 1H. -CONH), 10.6 (s, 1H, pyrimidine NH) ppm;

El IIS b / z (relative intensity): printed by the Central Standards Bureau of the Ministry of Economic Affairs and Consumer Cooperatives 514 01+, 10), 499 (31), 391 (10). 362 (10). 349 ¢ 30), 340 ( 25), 312 (55), 283 C7), 189 (21), 179 (31), 160 (30), 153 (100), 141 (65), 84 (80); HRMS Calcd. For C25H34N606: 514- 2539; Found 514.2535. The scale of this paper is applicable to China National Standards (CNS) 8 4 specifications (210XW7 public) 4t4Q9〇A7 B7 V. Description of the invention (21) Table 2e ^ _ [4- (4- (2-Amino -6-methyl-4 (3H) -oxopyriniidin-5-yl) PiPerazin ~ l_ yl) benzoyl) glutamic acid (9) mp: 265-266 ° C; IR (KBr): 3320, 2944, 2815, 1704, 1659 , 1606, 1510, 1231 c®-l; 1H NMR (400 MHz, DMSO-d6): δ 1.95-2.05 (m, 2H, CH3CH2CH2-), 2.12 (s, 3H, CH3-), 2.30-2.36 (t , J = 7.4 Hz, 2H, CH3CH2CH2-), 2.85-3.5 (m, 8H, piperazinyMO, 4.33-4.40 (td, J = 7.4 Hz, 7.68 Hz, 1H.-NHCH-), 6.29 Cbrs, 2H, -NH2 ), 6.90-7.00 &amp; 7.74-7.80 (ABq, J = 8.84 Hz, 4Ht Ph-H), 8.20-8.30 (d, J = 7.68 Hz, 1H, -CONH);

El MS m / z (relative intensity): 458 (M +, 70), 424 (65), 407 (70), 361 (100). 2-Anjino-6-methyl-5-C4- (4-nitrophenyl) piperazin -l-yl) pyrimidin-4 (3H) 'one (10) mp: 330 ° C; (deep); IR (KBr): 3350 (br), 3150, 2825, 2740, 1660, 1630, 1590. 1490, 1390, 1100 cm1; ^ NMR (80 MHz, DMSO-d6); δ 2.13 (s, 3H, CH3-), 2-90-3,20 (m, 4H, piperazinyl-H), 3.40-3.70 (id, 4H, piperazinyl-H), the size of the paper method is applicable to China National Standard (CNS) A4 (210X297 mm) 24 ---------- f-- (please read the note on the back of t * first) (Fill in this page again) -10 Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of Invention (22) Table 2 f 6.00-6.50 (brs, 2H, -NJ ^), 6.85-7.2 &amp; 7.89-8 : 2 (ABqf J = 9,36 Hz, 4H, Ph-H) ppra;

El MS b / z (relative intensity): 330 (M +, 85), 315 (45), 179 (20), 166 (18), 153 (100), 125 (25), 73 (20); HRMS Calcd. for C15H18O3N6: 330.1440; Found 330.1443. ί, N-Dibenzyl 4- (4- (2-Anjino-6-niethyl-4 (3H) -oxopyriiDidin-5-yI) pif) eraziii ~ ly]] beiizenesulf〇iiamicle ( 11) mp: 289-290 ° C; IR (KBr): 3400 (br), 3150 (br), 3060, 2920, 2850, 1670, 1650, 16 ^ 1590, 1510, 1315, 1307, 1150 cnj ~ i; 1H NMR (80 MHz, DMSO-dg): (5 2.14 (s, 3H, CH3-) f 2.89-3.19 (ffl, 4H, piperazinyl-H), 3,20-3.40 (ra, 4H, piperazinyl-H) , 4.22 (s, 4H, -CH2-Ph), 6.15-6.41 (brs, 2H, -NH2) f Printed by the Central Standard of the Ministry of Economy 扃 Employee Consumer Cooperative (Please read the precautions on the back before filling this page) 6.9- 7.8 (m, 14H, Ph-H) ppm;

El t ^ m / z (relative intensity): 544 (M +, 20), 529 (85), 421 (80), 379 (100), 153 (35), 91 (65);

Ana for C29H32N6O3S: Calcd. C, 63.95; H, 5.92; N, 15.43;

Found C, 63.60; fl, 5.91; N, 15.20. This paper size applies to China's national standard (CNS) A4jyU $ ((2 丨 0X297 mm) 25 A7 B7 g Printed by the Central Laboratories of the Ministry of Economic Affairs 42 ^ 090 5. Description of the invention (23) Table · 4- [4- (2-Amino-6-DiethyI-4 (3H) -〇x〇pyrimidin-5-yl) pif) erazin-l-rObenzenesulfonamide (12) rap: 300 ° C; (deep) IR (KBr): 3436, 2750 (br), 1651, 1596, 1117, 1110 curk 1H NMR (80MHz, DMS0-d6): 5 2.12 (s, 3H, CH3), 3.9 -3.5 (ra, 8H, piperazinyl-H), 6.27 (s, 2H, N§2), 6.9- 7.2 (m, 4H, Ph-H, S02NH2), 7.4-7.7 (d, 2H, J = 8.2 Hz , Ph-H) ppm;

El MS m / z (relative intensity): 364 (M +, 20), 349 (50), 153 (100) HRMS Calcd. For Ci5H2〇N6〇3S: 364.1317: Found 364.1313. 2-Amino-5-C4- ( 4-aminomethylphenyl) -piperazin-l-yl) 6 ~ iDethyl-3yrimidine-4-〇l (13) nip: 281-282.5 ° C; (deep); IR (KBr) · 3435, 3145, 2918, 1651, 1558, 1517 cur !; 1HNMR (80 MHz, D2〇 ™): δ 1.99 (s, 3H, CH3-), 2.90-3.50 (jb, 8H, piperazinyl-H), 3.86 (s, 2H, -CH2 NH2) , 7.1-7.4 (m, 4H, Ph-H) ppm; This paper is a standard for China National Standards (CNS) A4 (210X297 cm) 26 (Please read the precautions on the back before filling this page)

424090 Printed by the Consumers' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs Α7 Β7 V. Description of Invention (24) Table 2h

El MS m / z (relative intensity): 314 (M +, 52), 299 (90), 179 (21), 166 (30), 153 (100), 133 125 (40), 118 (30), 106 ( 42), 77 (20); HJ? HS for C16H22NgO: Calcd: 314.1855; Found: 314.1850. 4- [4- (2-Amino-4-hydroxy-6-methyl-pyriiDidin-5-yl) -piperazin-1 -yl) benzoic acid ethyl ester (14) nip: 330 ° C; (deep); rap: 290-291 ° C; IR (KBr): 3400 (br), 3180 (br), 2980, 2840, 1680, 1650 , 1610, 1380, 1330, 1275, 1235, 1190, 1160 cni-l; 1H NMR (80 MHz, DMS0 ~ d6): δ 2.15 (s, 3H, CH3-), 2.80- 3.60 (m, 8B, piperazinyl- H), 6.40-6.78 (brs, 2H, -NH2), 6.80- 7.10 (d, 2H, J = 8.84 Hz, Ph-H), 7.60-7.90 (d, 2H, J = 8.84 Hz, Ph-H) ppm; HRMS for C18% N503: Calcd 357.1800; Found: 357.1802. This paper size is applicable to national sample (CNS) A4 specification (210X297 mm) 27 (Please read the precautions on the back before filling in this I)

T

Claims (1)

6. Scope of patent application 1. A compound having the structure shown by formula (I) and a pharmaceutically acceptable salt thereof, wherein I is a hydrogen group or a Cm alkyl group, and R2 and R3 may respectively represent a hydrogen group and a Cm alkyl group; R4 And R5 may respectively represent fluorenyl, cyano, C〗 -3 alkanoyl, nitro (Nitro), and (2) -COOR8, where R8 is hydrogen, alkyl, and pharmaceutically acceptable metal ion, A pharmaceutically acceptable amine ion, (3) S02R12, wherein R12 represents a hydroxyl group, a C 6 alkyl group, or a C w alkoxy group; or -NR13 R14, wherein R 13 and R 14 may be a hydrogen group and C i_6 ------- --T- (Please read the notes on the back before filling in this page> Employees' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, Yin Fan2. For example, if the scope of the patent application is the first structural compound of formula (I) and its pharmaceutically acceptable salts, where K is hydrogen or alkynyl 114, 115 can represent ) Cyano, Cw alkanoyl, (2) -COOR8 where R8 is hydrogen, Cw alkyl, pharmaceutically acceptable amine ion, (3) S02R12 where R12 is -NR13R14 where R13R14 can be hydrogen, C, _6 alkyl, benzyl. 3. According to the scope of the patent application, the structural compound of formula (I) and its pharmaceutically acceptable salts, in which R1 is hydrogen or CU3, and R5 can respectively represent (1) hydrogen and C〗 -3. (Alkanoyl), this paper size applies to Chinese national standard (CNS &gt; A4 now (210X297 mm) 28 ABCD 42A090 6. Application scope (2) S02NR13R14 Rn, Ri4 can respectively represent hydrogen radical, c 4_A pharmaceutical composition for anti-brain cancer cells, which is selected from an effective dose of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the first patent application range, mixed with a pharmaceutically acceptable excipient 5. · —A method for preparing a structural compound as shown in formula (1) of the scope of patent application, which uses an organic solvent containing any of the experimental ingredients of sodium hydroxide, potassium hydroxide, sodium carbonate, and potassium carbonate. The formula (π) reacts with the formula (m) to form salts, if necessary, to form a pharmaceutically acceptable salt; and the organic solvent is selected from the group consisting of nitrogen nitrogen difluorenyl (DNF) difluorenyl oxygen sulfur compound (DMSO) Either (Please read the notes on the back before filling this page) (II) (ΠΙ) Order printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs 6. If the method of preparation of the scope of patent application for item 5 is prepared, it is the reaction of the formula (Π) with the formula I substitution (dish) Salts are formed; wherein R2 and R3 of formula (Π) represent a hydrogen group and a methyl group, respectively. The size of this paper is Chinese National Standard (CNS) A4 (210X297 mm) 29