2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-n-arylacetamide derivatives, processes for their preparation and use

Abstract

The present application relates to 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidine-1(2H)-yl)-N-arylacetamide derivatives and pharmaceutically acceptable salts thereof, pharmaceutical compositions and their use in the manufacture of antitumor drugs, as shown in the chemical structural formula I: wherein R is selected from methoxy, dimethoxy, trimethoxy, 2-hydroxy-3-methoxy, 2-hydroxy-4-methoxy, 2-hydroxy-5-methoxy, 2-hydroxy-6-methoxy, 3-hydroxy-2-methoxy, 3-hydroxy-4-methoxy, 3-hydroxy-5-methoxy, 3-hydroxy-6-methoxy, 4-hydroxy-2-methoxy, 4-hydroxy-3-methoxy, 4-hydroxy-3,5-dimethoxy, 2-hydroxy-3-ethoxy, 2-hydroxy-4-ethoxy, 2-hydroxy-5-ethoxy, 2-hydroxy-6-ethoxy, 3-hydroxy-2-ethoxy, 3-hydroxy-4-ethoxy, 3-hydroxy-5-ethoxy, 3-hydroxy-6-ethoxy, 4-hydroxy-2-ethoxy, 4-hydroxy-3-ethoxy or 4-hydroxy-3,5-diethoxy.

Description

Technical Field

[0001] This invention relates to a new class of compounds, their preparation methods and applications, specifically 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivatives, their preparation methods and their applications as antitumor drugs. Background Technology

[0002] Currently, the main treatment for malignant tumors is chemotherapy (drug therapy), supplemented by radiotherapy, biological therapy and immunotherapy, but no more effective treatment methods have been found to date.

[0003] 5-Fluorouracil is an antimetabolite drug. Its mechanism is that 5-fluorouracil and its derivatives bind to deoxyribonucleotides at the N1 position, inhibiting the function of thymine synthase, which leads to the ineffective synthesis of thymine deoxyribonucleotides, thereby interfering with DNA replication and synthesis. Because its chemical structure is similar to that of pyrimidine bases, which are essential metabolites for cell proliferation in the body.

[0004] Although 5-fluorouracil has wide clinical applications, its low cell selectivity, significant toxic side effects, and the close proximity of therapeutic and toxic doses cause considerable suffering for patients. To mitigate these drawbacks, 5-fluorouracil undergoes structural modification to optimize its molecular structure, reduce toxicity, improve selectivity, enhance in vivo metabolic kinetics, and increase in vivo stability.

[0005] In 2003, Dominguez et al. [Bioorganic & Medicinal Chemistry, 2003, 11(3):315–323] described a series of 5-fluorouracil prodrugs, among which compound A1 can induce morphological and phenotypic differentiation of rhabdomyosarcoma cells at 4.5 μM, which can serve as an alternative to selectively destroying undifferentiated cells.

[0006]

[0007] In 2007, Tian et al. [Molecules, 2007, 12(11): 2450–2457] synthesized six 5-fluorouracil analogs. Preliminary in vitro antitumor activity studies showed that they had certain cytotoxicity against B16, K562 and CHO cells. Some compounds could effectively inhibit the growth of tumor cells, among which compound A2 showed high inhibitory activity.

[0008]

[0009] In 2019, Liu et al. [MedChemComm, 2019, 10: 1370–1378] described a series of pentacyclic triterpenoid-5-fluorouracil conjugates and evaluated their antitumor activity. The results showed that compound A3 possessed certain antiproliferative activity, with an IC50 of 100% against multidrug-resistant cell lines A549 / T and Bel-7402 / FU. 50 The values ​​were 20.73 and 19.77 μmol / L, respectively.

[0010]

[0011] In 2020, Mustafa et al. [Systematic Review Pharmacy, 2020, 11(3): 482–489] described methyl cinnamate derivatives of (5-fluoro-2,4-dioxo-3,4-dihydropyrimidine-1(2H)-yl)cinnamate. In vitro anticancer activity tests revealed that some compounds exhibited inhibitory effects on the proliferation of MCF-7, HeLa, AMN3, and SKG cells. Among these, compound A4 showed the highest IC50 value. 50 The values ​​were 290, 327, 298 and 325 μmol / L, respectively.

[0012]

[0013] In 2021, Salerno et al. [Journal of Enzyme Inhibition and Medicinal Chemistry, 2021, 36(1): 1378–1386] synthesized compound A5, which interacts with a heme oxygenase 1 inhibitor. Preliminary cytotoxicity evaluation showed that compound A5 had an IC50 of 100% against human prostate cancer cells (DU145) and lung cancer cells (A549). 50 The values ​​were 46.93 and 1.45 μmol / L, respectively.

[0014] Summary of the Invention

[0015] The technical problem solved by the present invention is to provide a class of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivatives, their preparation methods, pharmaceutical compositions and uses.

[0016] To solve the technical problem of this invention, the present invention provides the following technical solution:

[0017] The first aspect of the present invention is to provide a class of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivatives as shown in structural formula I, and pharmaceutically acceptable salts thereof:

[0018]

[0019] Wherein, R is selected from: 2-methoxy, 3-methoxy, 4-methoxy, 2,3-dimethoxy, 2,4-dimethoxy, 2,5-dimethoxy, 2,6-dimethoxy, 3,4-dimethoxy, 3,5-dimethoxy, 2-hydroxy-3-methoxy, 2-hydroxy-4-methoxy, 2-hydroxy-5-methoxy, 2-hydroxy-6-methoxy, 3-hydroxy-2-methoxy, 3-hydroxy-4-methoxy, 3-hydroxy-5-methoxy, 3-hydroxy-6-methoxy, 4-hydroxy-2-methoxy Methoxy, 4-hydroxy-3-methoxy, 4-hydroxy-3,5-dimethoxy, 2-hydroxy-3-ethoxy, 2-hydroxy-4-ethoxy, 2-hydroxy-5-ethoxy, 2-hydroxy-6-ethoxy, 3-hydroxy-2-ethoxy, 3-hydroxy-4-ethoxy, 3-hydroxy-5-ethoxy, 3-hydroxy-6-ethoxy, 4-hydroxy-2-ethoxy, 4-hydroxy-3-ethoxy, 4-hydroxy-3,5-diethoxy, 2,3,4-trimethoxy, or 3,4,5-trimethoxy.

[0020] Further, preferred compounds are selected from: 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(2-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(3-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(4-methoxyphenyl)acetamide, N-(2,3-dimethoxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(2,4 ... 2H)-yl)acetamide, N-(2,5-dimethoxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(2,6-dimethoxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(3,4-dimethoxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(3,5-dimethoxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(2-hydroxy) 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(2-hydroxy-4-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(2-hydroxy-5-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(2-hydroxy-6-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(3-hydroxy-2-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(3-hydroxy-2-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(2-hydroxy-6-methoxyphenyl)acetamide, 1(2H)-yl)-N-(3-hydroxy-4-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(3-hydroxy-5-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(3-hydroxy-6-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(4-hydroxy-2-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(4-hydroxy-3-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(4-hydroxy-3-methoxyphenyl)acetamide, 2-(5-fluoro-2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(4-hydroxy-3,5-dimethoxyphenyl)acetamide, N-(3-ethoxy-2-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(3-ethoxy-2-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(4-ethoxy-2-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(4-ethoxy-2-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide N-(5-ethoxy-2-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(6-ethoxy-2-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(2-ethoxy-3-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(4 ...5-ethoxy-2-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3-hydroxyphenyl)-2-(5-fluoro-2 -3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(5-ethoxy-3-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(6-ethoxy-3-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(2-ethoxy-4-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(3-ethoxy-4-hydroxyphenyl)- 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, N-(3,5-diethoxy-4-hydroxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide, 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(2,3,4-trimethoxyphenyl)acetamide, or 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(3,4,5-trimethoxyphenyl)acetamide.

[0021] The second aspect of the present invention provides a method for preparing a 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivative, characterized in that its preparation reaction is as follows:

[0022]

[0023] Wherein, R is selected from: 2-methoxy, 3-methoxy, 4-methoxy, 2,3-dimethoxy, 2,4-dimethoxy, 2,5-dimethoxy, 2,6-dimethoxy, 3,4-dimethoxy, 3,5-dimethoxy, 2-hydroxy-3-methoxy, 2-hydroxy-4-methoxy, 2-hydroxy-5-methoxy, 2-hydroxy-6-methoxy, 3-hydroxy-2-methoxy, 3-hydroxy-4-methoxy, 3-hydroxy-5-methoxy, 3-hydroxy-6-methoxy, 4-hydroxy-2-methoxy Methoxy, 4-hydroxy-3-methoxy, 4-hydroxy-3,5-dimethoxy, 2-hydroxy-3-ethoxy, 2-hydroxy-4-ethoxy, 2-hydroxy-5-ethoxy, 2-hydroxy-6-ethoxy, 3-hydroxy-2-ethoxy, 3-hydroxy-4-ethoxy, 3-hydroxy-5-ethoxy, 3-hydroxy-6-ethoxy, 4-hydroxy-2-ethoxy, 4-hydroxy-3-ethoxy, 4-hydroxy-3,5-diethoxy, 2,3,4-trimethoxy, or 3,4,5-trimethoxy.

[0024] A third aspect of the present invention is to provide a pharmaceutical composition comprising the compound described in the first aspect and its pharmaceutically acceptable salt, wherein the pharmaceutical composition contains a therapeutically effective amount of the 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivative of the present invention and its pharmaceutically acceptable salt, and optionally a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier refers to a pharmaceutically acceptable carrier commonly used in the pharmaceutical field; the pharmaceutical composition can be prepared according to methods known in the art. Any dosage form suitable for human or animal use can be formulated by combining the compound of the present invention and its pharmaceutically acceptable salt with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants. The content of the compound of the present invention and its pharmaceutically acceptable salt in its pharmaceutical composition is typically 0.1% to 95% by weight.

[0025] The compounds of the present invention and their pharmaceutically acceptable salts or pharmaceutical compositions containing them can be administered in unit dose form via enteral or non-enteric routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0026] Dosage forms can be liquid, solid, or semi-solid. Liquid dosage forms can include solutions (including true solutions and colloidal solutions), emulsions (including o / w, w / o, and double emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can include tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, microcapsules, pellets, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can include ointments, gels, and pastes, etc.

[0027] The compounds of this invention and their pharmaceutically acceptable salts can be formulated into conventional formulations, as well as sustained-release formulations, controlled-release formulations, targeted formulations, and various microparticle delivery systems.

[0028] To formulate the compounds of the present invention and their pharmaceutically acceptable salts into tablets, a wide variety of excipients known in the art can be used, including diluents, binders, wetting agents, disintegrants, lubricants, and flow aids. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, isopropanol, etc.; binders can be starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfonate, etc.; lubricants and flow aids can be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0029] Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0030] To formulate the drug delivery unit into capsules, the active ingredient, the compound of the present invention, and its pharmaceutically acceptable salts, can be mixed with a diluent and a gliding agent, and the mixture can be placed directly into hard or soft capsules. Alternatively, the active ingredient, the compound of the present invention, and its pharmaceutically acceptable salts, can be first formed into granules or microspheres with a diluent, binder, and disintegrant, and then placed into hard or soft capsules. The diluents, binders, wetting agents, disintegrants, and gliding agents used to prepare tablets of the compound of the present invention and its pharmaceutically acceptable salts can also be used to prepare capsules of the compound of the present invention and its pharmaceutically acceptable salts.

[0031] To prepare the compounds of this invention and their pharmaceutically acceptable salts into injectable formulations, water, ethanol, isopropanol, propylene glycol, or mixtures thereof can be used as solvents, with the addition of appropriate amounts of commonly used solubilizers, co-solvents, pH adjusters, and osmotic pressure regulators. Solubilizers or co-solvents may include poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; pH adjusters may include phosphates, acetates, hydrochloric acid, sodium hydroxide, etc.; and osmotic pressure regulators may include sodium chloride, mannitol, glucose, phosphates, acetates, etc. If preparing lyophilized powder injections, mannitol, glucose, etc., may also be added as supporting agents.

[0032] In addition, colorants, preservatives, flavorings, tasters or other additives may be added to pharmaceutical preparations if necessary.

[0033] To achieve the purpose of medication and enhance the therapeutic effect, the drug or drug composition of the present invention can be administered using any known method of administration.

[0034] The fourth aspect of the present invention is to provide the use of the 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivatives of the present invention and their pharmaceutically acceptable salts, as well as the pharmaceutical compositions of the third aspect, in the preparation of antitumor drugs.

[0035] Beneficial technical effects:

[0036] The 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivative of the present invention is a novel class of compounds with antitumor activity. Detailed Implementation

[0037] The following examples are intended to illustrate the invention and not to further limit it.

[0038] Example 1

[0039] A 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivative has the following molecular formula:

[0040]

[0041] Wherein, R is selected from: 2-methoxy, 3-methoxy, 4-methoxy, 2,3-dimethoxy, 2,4-dimethoxy, 2,5-dimethoxy, 2,6-dimethoxy, 3,4-dimethoxy, 3,5-dimethoxy, 2-hydroxy-3-methoxy, 2-hydroxy-4-methoxy, 2-hydroxy-5-methoxy, 2-hydroxy-6-methoxy, 3-hydroxy-2-methoxy, 3-hydroxy-4-methoxy, 3-hydroxy-5-methoxy, 3-hydroxy-6-methoxy, 4-hydroxy-2-methoxy Methoxy, 4-hydroxy-3-methoxy, 4-hydroxy-3,5-dimethoxy, 2-hydroxy-3-ethoxy, 2-hydroxy-4-ethoxy, 2-hydroxy-5-ethoxy, 2-hydroxy-6-ethoxy, 3-hydroxy-2-ethoxy, 3-hydroxy-4-ethoxy, 3-hydroxy-5-ethoxy, 3-hydroxy-6-ethoxy, 4-hydroxy-2-ethoxy, 4-hydroxy-3-ethoxy, 4-hydroxy-3,5-diethoxy, 2,3,4-trimethoxy, or 3,4,5-trimethoxy.

[0042] Example 2

[0043] Preparation of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetic acid

[0044]

[0045] 62.5 mmol of potassium hydroxide was dissolved in 15 mL of aqueous solution. The solution was stirred at room temperature until dissolved, followed by the addition of 15.4 mmol of 5-fluorouracil. After stirring until dissolved, the solution was heated to 80 °C, and 21.6 mmol of bromoacetic acid was slowly added dropwise. The reaction was continued with stirring for 11 h, and the reaction process was monitored by TLC. The reaction solution was cooled to room temperature, and the pH was adjusted to 2-3 with concentrated hydrochloric acid. Water was removed from the reaction solution by rotary evaporation under reduced pressure. The crude product was recrystallized from an appropriate amount of ethanol, filtered, and dried to give 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetic acid, a white solid, mp >250 °C, with a yield of 97%.

[0046] Example 3

[0047] Preparation of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(3-methoxyphenyl)acetamide (M1)

[0048]

[0049] 1.0 mmol of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetic acid, 1.2 mmol of 3-methoxyaniline, 1.2 mmol of HOBt, 0.2 mmol of DMAP, and 2.0 mmol of EDCI were dissolved in 10 mL of DMF and reacted with stirring at room temperature for 48 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was quenched with saturated NaHCO3 solution, diluted with EtOAc, and extracted with EtOAc (2×50 mL). The combined organic extracts were dried over anhydrous Na2SO4, filtered, concentrated by rotary evaporation under reduced pressure, purified by silica gel column chromatography, and dried to give 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(3-methoxyphenyl)acetamide (M1), a white powder, mp 220–221 °C, yield 91%. 1 H NMR(500 MHz, DMSO-d6) δ: 11.88 (s, 1H, NH), 10.31 (s, 1H, CONH), 8.09 (d, J = 6.8Hz, 1H, C4HN2), 7.29 (s, 1H, C6H4), 7.22 (m, 1H, C6H4), 7.08 (d, J = 8.1 Hz,1H, C6H4), 6.65 (d, J = 8.1 Hz, 1H, C6H4), 4.49 (s, 2H, COCH2), 3.72 (s, 3H,OCH3), 13 C NMR (126 MHz, DMSO-d6) δ: 165.38, 158.98, 157.64, 157.43, 150.59,139.70, 131.28, 129.16, 111.30, 109.12, 105.15, 54.97, 50.21.

[0050] Example 4

[0051] Preparation of N-(3,4-dimethoxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide (M2)

[0052]

[0053] 1.0 mmol of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetic acid, 1.2 mmol of 3,4-dimethoxyaniline, 1.2 mmol of HOBt, 0.2 mmol of DMAP, and 2.0 mmol of EDCI were dissolved in 10 mL of DMF and reacted with stirring at room temperature for 48 h. The reaction was monitored by TLC. After the reaction was complete, it was quenched with saturated NaHCO3 solution, and the reaction mixture was diluted with EtOAc and extracted with EtOAc (2×50 mL). The combined organic extracts were dried on anhydrous Na2SO4, filtered, concentrated by rotary evaporation under reduced pressure, purified by silica gel column chromatography, and dried to give N-(3,4-dimethoxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide (M2), a white powder, mp 245~246℃, yield 92%. 1 H NMR (500 MHz, DMSO-d6) δ: 11.80 (s, 1H, NH), 10.37 (s, 1H, CONH), 8.11 (d, J = 6.7Hz, 1H, C4HN2), 7.35 (s, 1H, C6H3), 7.06 (d, J = 8.7 Hz, 1H, C6H3), 6.89 (d, J= 8.7 Hz, 1H, C6H3), 4.49 (s, 2H, COCH2), 3.71 (s, 3H, OCH3), 3.71 (s, 3H,OCH3); 13 C NMR (126 MHz, DMSO-d6) δ: 164.86, 157.65, 157.45, 156.06, 150.19,144.93, 132.23, 130.47, 112.07, 110.52, 104.18, 55.69, 55.32, 50.11.

[0054] Example 5

[0055] Preparation of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(4-methoxyphenyl)acetamide (M3)

[0056]

[0057] 1.0 mmol of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetic acid, 1.2 mmol of 4-methoxyaniline, 1.2 mmol of HOBt, 0.2 mmol of DMAP, and 2.0 mmol of EDCI were dissolved in 10 mL of DMF and reacted with stirring at room temperature for 48 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was quenched with saturated NaHCO3 solution, diluted with EtOAc, and extracted with EtOAc (2 × 50 mL). The combined organic extracts were dried over anhydrous Na2SO4, filtered, concentrated by rotary evaporation under reduced pressure, purified by silica gel column chromatography, and dried to give 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(4-methoxyphenyl)acetamide (M3), a white powder, mp 237–238 °C, yield 89%. 1 H NMR (400 MHz, DMSO-d6) δ: 11.89 (s, 1H, NH), 10.12 (s, 1H, CONH), 8.09 (d, J = 6.8Hz, 1H, C4HN2), 7.52~7.43 (m, 2H, C6H4), 6.93~6.85 (m, 2H, C6H4), 4.46 (s, 2H,COCH2), 3.72 (s, 3H, OCH3).

[0058] Example 6

[0059] Antitumor activity of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivatives

[0060] 1. Principle of antitumor activity experiment

[0061] In living cells, mitochondria contain succinate dehydrogenase, which reduces exogenous MTT to insoluble blue-purple crystalline formazan, which then deposits within the cell. Dead cells lack this ability. Dimethyl sulfoxide (DMSO) can dissolve the purple crystals in cells; the absorbance at 490 nm using an enzyme-linked immunosorbent assay (ELISA) scanner indirectly reflects cell number. Within a certain cell number range, the amount of MTT crystals formed is directly proportional to the cell number.

[0062] 2. Experimental methods for antitumor activity

[0063] Cell lines: 4T1 (mouse breast cancer cells) and TPC-1 (human papilloma thyroid cancer cells), donated by the Laboratory of Modern Engineering and Applied Science, Nanjing University.

[0064] Reagents: Thiazol blue (MTT), RPMI 1640 culture medium containing antibiotics (Nanjing Kaiji Biotechnology Development Co., Ltd.), fetal bovine serum (Zhejiang Tianhang Biotechnology Co., Ltd.), trypsin (Nanjing Kaiji Biotechnology Development Co., Ltd.), 96-well culture plate (Wuxi Nais Life Science Co., Ltd.), dimethyl sulfoxide (Sigma (Shanghai) Co., Ltd.).

[0065] Instruments: 1300 series A2 biosafety cabinet (Thermo Scientific, USA), CLM-170B-8-CN CO2 incubator (ESCO), BDS400 inverted microscope (Chongqing Aote Optical Instrument Co., Ltd.), Multiskan GO microplate reader (Thermo Scientific, USA), ultrapure water preparation system (Milli-Q, USA).

[0066] Experimental Procedure: The samples were tested against 4T1 (mouse breast cancer cells) and TPC-1 (human papilloma-thyroid cancer cells). In one experiment, seven concentration gradients were set for each sample (2500 ng / L, 1250 ng / L, 625 ng / L, 312.5 ng / L, 156 ng / L, 78 ng / L, and 39 ng / L). The drugs were dissolved in DMSO and incubated for 72 hours. Four parallel samples were prepared for each concentration, and each experiment was repeated in triplicate. A blank control was used for reference. The OD value of each well was measured using a microplate reader at a wavelength of 490 nm.

[0067] 3. Samples to be tested: Compounds 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(3-methoxyphenyl)acetamide (M1) and N-(3,4-dimethoxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide (M2).

[0068] 4. Evaluation of antitumor activity

[0069] 4T1 (mouse breast cancer cells) and TPC-1 (human papillomathyroid carcinoma cells) cells were incubated with DMSO to dissolve the drug. After incubation for 72 hours, cell viability was detected by the MTT assay. The results showed that compounds 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-(3-methoxyphenyl)acetamide (M1) and N-(3,4-dimethoxyphenyl)-2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide (M2) had significant cytotoxicity against cancer cells. Compounds M1 and M2 showed high IC50 values ​​against 4T1 (mouse breast cancer cells) and TPC-1 (human papillomathyroid carcinoma cells). 50 The values ​​are listed in Table 1.

[0070] Table 1. IC50 of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivatives on 4T1 and TPC-1 cells. 50

[0071]

[0072]

[0073] Activity test results showed that the 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivative had good inhibitory activity against breast cancer 4T1 cells and human papillomatous thyroid carcinoma TPC-1 cells, and could be used to prepare antitumor drugs.

Claims

1. A class of 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivatives of chemical structural formula I and their pharmaceutically acceptable salts: in, R is selected from: 3-methoxy, 2,3-dimethoxy, 2,4-dimethoxy, 2,5-dimethoxy, 2,6-dimethoxy, 3,4-dimethoxy, 3,5-dimethoxy, 2-hydroxy-3-methoxy, 2-hydroxy-4-methoxy, 2-hydroxy-5-methoxy, 2-hydroxy-6-methoxy, 3-hydroxy-2-methoxy, 3-hydroxy-4-methoxy, 3-hydroxy-5-methoxy, 3-hydroxy-6-methoxy, 4-hydroxy-2-methoxy, 4-hydroxy -3-methoxy, 4-hydroxy-3,5-dimethoxy, 2-hydroxy-3-ethoxy, 2-hydroxy-4-ethoxy, 2-hydroxy-5-ethoxy, 2-hydroxy-6-ethoxy, 3-hydroxy-2-ethoxy, 3-hydroxy-4-ethoxy, 3-hydroxy-5-ethoxy, 3-hydroxy-6-ethoxy, 4-hydroxy-2-ethoxy, 4-hydroxy-3-ethoxy, 4-hydroxy-3,5-diethoxy, 2,3,4-trimethoxy, or 3,4,5-trimethoxy.

2. The use of the 2-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-N-arylacetamide derivative of claim 1 and its pharmaceutically acceptable salt in the preparation of antitumor drugs.

3. A pharmaceutical composition comprising at least one compound of claim 1 and a pharmaceutically usable carrier.

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