CN111249283A - Pyrimidine derivatives having anticancer effect - Google Patents
Pyrimidine derivatives having anticancer effect Download PDFInfo
- Publication number
- CN111249283A CN111249283A CN201811458195.9A CN201811458195A CN111249283A CN 111249283 A CN111249283 A CN 111249283A CN 201811458195 A CN201811458195 A CN 201811458195A CN 111249283 A CN111249283 A CN 111249283A
- Authority
- CN
- China
- Prior art keywords
- substituted
- unsubstituted
- alkyl
- halogen
- compound
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
- C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
- C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
- C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
- C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
- C07D239/47—One nitrogen atom and one oxygen or sulfur atom, e.g. cytosine
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
- C07D239/70—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings condensed with carbocyclic rings or ring systems
- C07D239/72—Quinazolines; Hydrogenated quinazolines
- C07D239/86—Quinazolines; Hydrogenated quinazolines with hetero atoms directly attached in position 4
- C07D239/88—Oxygen atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a pyrimidine derivative with an anticancer effect, and belongs to the field of chemical medicine. The invention provides a compound shown as a formula I or pharmaceutically acceptable thereofUse of a salt for the manufacture of a medicament for the treatment and/or prevention of cancer. Biological experiments show that the series of compounds have good inhibition effect on histone arginine demethylase, and IC of partial compounds50At micromolar level, the compound shows good inhibition effect on various clinical common tumor cell strains, and IC50Are all less than 1 mu M. The compound of the invention also shows good effect in vivo and can obviously inhibit the tumor growth in the body of a mouse. In a word, the invention provides a new effective selection for developing a new generation of targeted drugs for tumor therapy, and has good application prospect.
Description
Technical Field
The invention relates to a pyrimidine derivative with an anticancer effect, and belongs to the field of chemical medicine.
Background
Epigenetics is a subject of study on genetic changes in gene expression without changes in the nucleotide sequence of the gene. As with traditional genetics, epigenetics influences fundamental physiological functions related to human life development by regulating the process of gene transcription in organisms. Common epigenetic modification mechanisms mainly include: DNA modification, histone modification, RNA modification, chromatin remodeling, non-coding RNA, and the like. Histone modification is the most widely studied epigenetic modification mechanism at present, and the regulatory factors can be divided into three major categories according to the difference of the histone modification regulatory factors: write factor (Writer), erase factor (Eraser), and read factor (Reader). Histone demethylases are an important class of erase factors that are primarily involved in regulating the methylation balance of histones in the organism. Histone demethylases can be divided into two families according to the mechanism of action: one class is Flavin Adenine Dinucleotide (FAD) -dependent lysine-specific demethylases, including LSD1 and LSD 2; another class is the histone demethylases containing the jumonji domain (JmjC).
JMJD6 is a histone arginine demethylase containing JmjC domain, which catalyzes the demethylation modification of the methyl group at the arginine residue R2 on histone H3 and the arginine residue R3 on histone H4. Recent studies show that the error regulation of JMJD6 is closely related to the occurrence, development, metastasis and drug resistance of various human malignant tumors such as breast cancer, non-small cell lung cancer (NCSLC), melanoma, oral cancer, glioma, ovarian cancer, pancreatic cancer and the like. For example, JMJD6 can modulate the transcriptional pause-release process within glioma cells, thereby affecting cell survival; inhibition of the activity of JMJD6 is effective in inhibiting proliferation, metastasis and invasion of glioma cells.
The 2018 global cancer statistics report shows that the incidence and mortality of ovarian cancer are ranked 8 th of the incidence and mortality of female tumors. Currently, clinical therapeutic drugs for ovarian cancer mainly include the following two types: (1) cytotoxic drugs represented by cisplatin, paclitaxel, gemcitabine and the like have poor selectivity, high toxicity and large side effect after use; (2) targeted PARP inhibitors represented by olaparib, rukapanib, and nilapanib. Although PARP inhibitors provide a revolutionary breakthrough in the treatment of ovarian cancer, the problem of drug resistance of PARP inhibitors has plagued clinical treatment of ovarian cancer. Therefore, the micromolecule inhibitor with high activity, low toxicity and effective in vivo brand new target is developed, and a brand new treatment means is provided for treating multiple malignant tumors such as ovarian cancer, pancreatic cancer and the like, so that the micromolecule inhibitor has important significance in clinic.
Disclosure of Invention
The present invention aims to provide a pyrimidine derivative having an anticancer effect.
The invention provides an application of a compound shown as a formula I or a pharmaceutically acceptable salt thereof in preparing a medicament for treating and/or preventing cancer:
wherein R is1、R2Independently selected from H, halogen, substituted or unsubstituted alkyl, or, R1And R2Linked to form a substituted or unsubstituted aryl group;
R3selected from substituted or unsubstituted aryl;
R4selected from H, substituted or unsubstituted alkyl, or, R4And R3Joined to form an alicyclic ring.
Further, R1、R2Independently selected from H, halogen, unsubstituted C1-C6 alkyl or halogen substituted C1-C6 alkyl, or R1And R2Linked to form a substituted or unsubstituted 5-to 6-membered aryl group.
Preferably, R1、R2Independently selected from H, halogen, unsubstituted C1-C3 alkyl or halogen substituted C1-C3 alkyl, or R1And R2Linked to form a substituted or unsubstituted phenyl group.
Preferably, R1、R2Independently selected from H or methyl, or, R1And R2Joined to form an unsubstituted phenyl group.
Preferably, R1Selected from H or methyl, R2Is H, or R1And R2Joined to form an unsubstituted phenyl group.
Further preferably, R1Is methyl, R2Is H.
Further, R3Selected from substituted or unsubstituted 5-to 14-membered aryl.
Preferably, R3Selected from substituted or unsubstituted 5-6 membered aryl.
Preferably, the aryl group contains 0 to 2 heteroatoms selected from nitrogen, oxygen or sulfur.
Preferably, R3Selected from substituted or unsubstituted furyl, pyrrolyl, thienyl, pyrazolyl, isoxazolyl, phenyl or pyridyl.
Preferably, the substituted aryl group contains at least one substituent selected from the group consisting of: hydroxyl, cyano, substituted or unsubstituted alkyl, halogen, substituted or unsubstituted amino, nitro.
Preferably, the substituted aryl group contains at least one substituent selected from the group consisting of: hydroxyl, cyano, unsubstituted C1-C6 alkyl, halogen substituted C1-C6 alkyl, halogen, amino, C1-C6 alkyl substituted amino and nitro.
Preferably, the substituted aryl group contains at least one substituent selected from the group consisting of: hydroxyl, cyano, unsubstituted C1-C6 alkyl, halogen substituted C1-C6 alkyl, halogen, amino, C1-C6 alkyl substituted amino and nitro.
Preferably, the substituted aryl group contains at least one substituent selected from the group consisting of: hydroxyl, cyano, methyl, fluorine, chlorine, amino, diethylamino and nitro.
Preferably, R3Selected from:
further, R4Selected from H, substituted or unsubstituted C1-C6 alkyl.
Preferably, R4Selected from H, unsubstituted C1-C3 alkyl or hydroxyl substituted C1-C3 alkyl. Go to
Preferably, R4Is selected from H or-CH2OH。
Further, R4And R3Are connected to form a 5-6 membered alicyclic ring.
Preferably, R4And R3Are linked to form a 5-membered alicyclic ring.
Further, R4Is H.
Further, the compound is selected from:
further, the cancer is ovarian cancer or pancreatic cancer.
Further, the drug is a histone demethylase inhibitor drug.
Preferably, the drug is a histone arginine demethylase JMJD6 inhibitor drug.
Furthermore, the medicine is a preparation prepared by taking the compound or the pharmaceutically acceptable salt thereof as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients.
Preferably, the formulation is an oral formulation or an injectable formulation.
Definition of terms:
the compounds and derivatives provided by the present invention may be named according to the IUPAC (international union of pure and applied chemistry) or CAS (chemical abstracts service, Columbus, OH) naming system.
The term "alkyl" is a radical of a straight or branched chain saturated hydrocarbon group. C1~C6Examples of alkyl groups include, but are not limited to, methyl (C)1) Ethyl (C)2) N-propyl (C)3) Isopropyl (C)3) N-butyl (C)4) Tert-butyl (C)4) Sec-butyl (C)4) Isobutyl (C)4) N-pentyl group (C)5) 3-pentyl radical (C)5) Pentyl group (C)5) Neopentyl (C)5) 3-methyl-2-butyl (C)5) Tert-amyl (C)5) And n-hexyl (C)6)。
The term "aryl" refers to a group of a 4n +2 aromatic ring system with or without heteroatoms in the aromatic ring system, wherein the heteroatoms are selected from nitrogen, oxygen and/or sulfur.
The term "alicyclic" refers to a saturated or partially unsaturated cyclic hydrocarbon group.
The term "pharmaceutically acceptable" means that the carrier, cargo, diluent, adjuvant, and/or salt formed is generally chemically or physically compatible with the other ingredients comprising a pharmaceutical dosage form and physiologically compatible with the recipient.
The term "pharmaceutically acceptable salts" refers to acid and/or base salts of the compounds of the present invention with inorganic and/or organic acids and bases, and also includes zwitterionic salts (inner salts), and also includes quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly in the final isolation and purification of the compounds. The compound may be obtained by appropriately (e.g., equivalent) mixing the above compound with a certain amount of an acid or a base. These salts may form precipitates in the solution which are collected by filtration, or they may be recovered after evaporation of the solvent, or they may be prepared by reaction in an aqueous medium followed by lyophilization. The salt in the invention can be hydrochloride, sulfate, citrate, benzene sulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.
The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.
Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or solubilizers, for example, starch, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.
Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.
Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like.
In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.
Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.
Dosage forms for topical administration of the compounds of the present invention include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.
The pharmaceutically acceptable auxiliary material of the invention refers to a substance contained in a dosage form except for an active ingredient.
The pharmaceutically acceptable auxiliary components have certain physiological activity, but the addition of the components does not change the dominant position of the pharmaceutical composition in the disease treatment process, but only plays auxiliary effects, and the auxiliary effects are only the utilization of the known activity of the components and are auxiliary treatment modes which are commonly used in the field of medicine. If the auxiliary components are used in combination with the pharmaceutical composition of the present invention, the protection scope of the present invention should still be included.
The invention provides a pyrimidine derivative with anticancer effect, which shows good tumor inhibition capability in vivo and in vitro. The invention has the beneficial effects that: firstly, the invention provides a novel histone arginine demethylase inhibitor which shows good inhibition effect on the in vitro enzyme level and the IC of partial compounds50(half maximal inhibitory concentration) on the micromolar scale; secondly, the compound shows good inhibition effect on a plurality of clinical common tumor cell strains in vitro, and IC50Are all less than 1 mu M; thirdly, the compound of the invention also shows good effect in vivo and can obviously inhibit the tumor growth in the body of a mouse. In a word, the invention provides a new effective selection for developing a new generation of targeted drugs for tumor therapy, and has good application prospect.
Drawings
FIG. 1 is a graph showing the results of immunoblotting for Compound 11 in example 4;
FIG. 2 is a graph of in vivo tumor suppression of Compound 11 of example 5;
FIG. 3 is a graph showing the change in body weight of mice in example 5;
fig. 4 is a graph of binding values of compound 11 to JMJD6 protein at each concentration in example 6.
Detailed Description
The raw materials and equipment used in the embodiment of the present invention are known products and obtained by purchasing commercially available products.
The invention provides an application of a compound shown as a formula I or a pharmaceutically acceptable salt thereof in preparing a medicament for treating and/or preventing cancer:
wherein R is1、R2Independently selected from H, halogen, substituted or unsubstituted alkyl, or, R1And R2Linked to form a substituted or unsubstituted aryl group;
R3selected from substituted or unsubstituted aryl;
R4selected from H, substituted or unsubstituted alkyl, or, R4And R3Joined to form an alicyclic ring.
As for the pyrimidine derivative containing the hydrazone functional group, the reports of the prior art on the pharmacological activity are limited to anti-inflammatory and antibacterial effects, and the invention creatively discovers that the compound also has obvious anticancer activity and provides a new choice for the development of clinical anticancer drugs.
EXAMPLE 12 preparation of (2- (2-hydroxybenzylidene) hydrazino) pyrimidin-4-ol (Compound 1)
A25 mL round bottom flask was charged with 2-hydrazinopyrimidin-4-ol (53mg, 0.5mmol) and 2-hydroxybenzaldehyde (106. mu.L, 1mmol) and 5mL methanol and refluxed at 65 ℃ overnight. The next day, the mixture is cooled to room temperature, filtered, the filter cake is washed by 10mL of absolute ethyl alcohol and 20mL of absolute ethyl ether in sequence, and dried to obtain 98mg of off-white solid with the yield of 85.2%.
1H NMR(400MHz,DMSO-d6)δ11.61(s,2H),9.01(s,1H),7.70(dd,J=7.7,1.5Hz,1H),7.43–7.37(m,2H),6.98(t,J=7.6Hz,3H),5.81(d,J=7.6Hz,1H).
ESI-ms(m/z):231.1[M+H]+
Compound 2-compound 24 can be prepared according to the preparation method of compound 1 by reacting different 2-hydrazinopyrimidine derivatives with the corresponding aldehydes or ketones, and the structure and characterization data are shown in table 1:
TABLE 1 structures of Compound 2-Compound 24,1H NMR and ESI-ms
Example 2 inhibition of histone demethylases by Compounds of the invention
Purpose of the experiment: assaying the inhibitory Activity of the Compounds of the present invention on histone demethylase in vitro Using Succinate-GloTMThe JmjC Demethyl laser/HydroxylaseAssay (Promega) kit tests the in vitro inhibitory activity of the compounds on the arginine Demethylase JMJD 6.
Experimental principle during histone demethylation, the cofactor α -ketoglutarate is reduced to succinate, if histone demethylase activity is inhibited, α -ketoglutarate is not reduced, therefore, the activity of histone demethylase can be indirectly reflected by determining the concentration of succinate.
The experimental method comprises the following steps: 2.5 mu L of small molecule solution with a certain concentration and 2.5 mu L of JMJD6 protein solution are added into a 384-well plate, the concentration of the JMJD6 protein solution is 2.2mg/mL, and after incubation for 10min at room temperature, 5 mu L of target protein substrate (Luc7like2 protein 267-274 amino acid polypeptide) solution is added. The components in the reaction system are mixed evenly and reacted for 8 hours at room temperature. Followed by addition of Succinate-GloTMReagent I (from Succinate-Glo) in JmjC Demethyl/Hydroxylase Assay kitTMBuffer solution, acetoacetyl coenzyme A and Succinate-GloTMThe solutions are mixed in proportion). After incubation for 1h at room temperature, reagent II from the kit (mixed proportionally with ATP detection buffer and ATP detection substrate) was added. Incubate at room temperature for 10min, and detect absorbance using a multifunctional microplate reader.
Inhibition (%) ═ 100% (control-drug treated group)/(control-blank control)%
Finally, the half Inhibitory Concentration (IC) was obtained by fitting with Graphpadprism software50)。
Through the above experimental method, the in vitro inhibitory activity of the compound of the present invention on arginine demethylase JMJD6 was tested.
Table 2 shows the in vitro inhibitory activity of the partial compounds of the present invention on histone arginine demethylase JMJD6, wherein A represents < 1. mu.M, B represents 1. mu.M-10. mu.M, C represents 10. mu.M-100. mu.M, and D represents > 100. mu.M.
TABLE 2 IC inhibition of JMJD6 by partial compounds of the invention50Value of
Compound numbering | IC50 | Compound numbering | IC50 |
1 | B | 13 | B |
2 | B | 14 | B |
3 | B | 15 | B |
4 | C | 16 | B |
5 | C | 17 | B |
6 | B | 18 | B |
7 | A | 19 | C |
8 | B | 20 | B |
9 | B | 21 | B |
10 | B | 22 | B |
11 | A | 23 | B |
12 | B | 24 | B |
As can be seen from the above table, the compound of the invention shows good inhibition effect on histone arginine demethylase JMJD6, IC50Values are on the micromolar scale.
EXAMPLE 3 inhibition of human tumor cell proliferation by Compound 11
Purpose of the experiment: the in vitro human tumor Cell proliferation inhibition activity of the compound of the invention is detected, and the Cell Counting kit 8 (CCK 8) method is adopted to test the inhibition activity of the tested compound on different tumor Cell strains. IC for tumor cell inhibitory Activity of test Compounds50(median inhibitory concentration) is shown. IC (integrated circuit)50Values can be obtained by calculating the inhibition rate of the test compound on tumor cells at a range of different concentrations.
The experimental principle is as follows: WST-8(2- (2-Methoxy-4-nitrophenyl) -3- (4-nitrophenyl) -5- (2, 4-disulfobenzene) -2H-tetrazole monosodium salt) can be reduced by dehydrogenase in mitochondria to generate a highly water-soluble orange-yellow formazan product (formazan) in the presence of an electron coupling carrier 1-Methoxy PMS, wherein the shade of color is in direct proportion to cell proliferation and in inverse proportion to cytotoxicity. The absorbance value is measured at a wavelength of 450nm by using a microplate reader, and the number of living cells is indirectly reflected.
The experimental method comprises the following steps: adjusting the cell concentration to 1-2 × 10 by using a whole cell culture solution4Cell suspension/mL, seeded into 96-well plates at 100. mu.l cell suspension per well, and cultured overnight. The next day, test compounds were added to the plates at different gradient concentrations, with a negative control group containing no drug and an equal volume of solvent control group, with a DMSO concentration of 0.1%, with 3 multiple wells per dose group, at 37 ℃ and 5% CO2Culturing under the condition. After 72 hours, 10. mu.L of CCK solution was added. Culture 1After-4 h, 150. mu.L of dimethyl sulfoxide (DMSO) was added to each well, mixed by shaking for 15min, and the absorbance was measured with a multifunctional microplate reader (. lamda. 450 nm).
Inhibition (%) ═ 100% (control-drug treated group)/(control-blank control)%
Finally, the half Inhibitory Concentration (IC) was obtained by fitting with Graphpadprism software50)。
Table 3 shows the proliferation inhibitory activity of Compound 11 against a variety of human tumor cell lines.
TABLE 3 IC inhibition of proliferation of Compound 11 by various human tumor cell lines50Value of
Cell line | Tumor type | IC50(μM) |
SK-OV-3 | Human ovarian cancer cell | 0.362 |
BxPC-3 | Human in situ pancreatic adenocarcinoma cells | 0.336 |
Capan-2 | Human pancreatic cancer cell | 0.889 |
Panc-1 | Human pancreatic cancer cell | 0.667 |
AsPC-1 | Human metastatic pancreatic adenocarcinoma cells | 0.262 |
MiaPaCa-2 | Human pancreatic cancer cell | 0.331 |
CFPAC-1 | Human pancreatic cancer cell | 0.541 |
The experimental result shows that the compound 11 has obvious inhibition effect on the proliferation of various human tumor cell strains in vitro.
EXAMPLE 4 intracellular inhibition of histone demethylases by Compound 11
Purpose of the experiment: and (3) detecting whether the compound can inhibit methylated arginine from carrying out demethylation modification in cells.
The experimental principle is as follows: the histone arginine demethylase JMJD6 can catalyze H3R2Me2 to carry out demethylation modification. The higher the activity of JMJD6, the lower the intracellular level of H3R2Me 2. The activity of JMJD6 can be indirectly reflected by measuring the content of H3R2Me2 in cells.
The experimental procedure after digesting SK-OV-3 cells from a petri dish with pancreatin, washing the pancreatin with PBS buffer (phosphate buffered saline solution), centrifugation (room temperature, 2000rpm, 5min) followed by lysis of the cells on ice with a RIPA lysis buffer containing the protease inhibitor Cocktail for 30min, during which time the supernatant is constantly aspirated with an insulin needle, vortexed, collected, mixed with SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel) protein loading buffer and heated on a 100 ℃ metal bath for 5min, the protein sample is electrophoretically separated and transferred to a PVDF (polyvinylidene fluoride) membrane, after blocking for 2H with 5% degreasing, incubated with 1:1000 diluted antibodies and PVDF membrane overnight at 4 ℃, washed with anti-H3R2Me2 antibody (arginine R2 on dimethylated histone H3), the methylation level of histone H3R2 is detected, while using anti-I- β -actin antibody (goat anti-actin antibody) is further washed with goat anti-immunoglobulin antibody labeled with goat anti-rabbit immunoblotting reference antibody, goat anti-goat-albumin antibody (goat anti-rabbit-goat anti-immunoglobulin antibody) and goat anti-goat-rabbit anti-immunoglobulin antibody are further labeled by a luminescence method, and labeled with anti-goat anti-rabbit antibody.
The immunoblotting results of Compound 11 were shown in FIG. 1.
The experimental result shows that the compound 11 can increase the dimethylation level of an arginine residue R2 on the histone H3 in an ovarian cancer SK-OV-3 cell strain in a dose-dependent manner, which indicates that the compound can inhibit the activity of JMJD6 in the SK-OV-3 cell strain, and further prevent JMJD6 from carrying out demethylation modification on H3R2Me 2.
EXAMPLE 5 in vivo antitumor Effect of Compound 11
Purpose of the experiment: the in vivo antitumor effect of the compounds of the present invention was examined. NOD-SCID mouse subcutaneous human ovarian cancer model was used in this experiment to test the in vivo anti-tumor activity of compound 11. The cell strain is human ovarian cancer cell strain SK-OV-3.
The experimental method comprises the following steps: culturing human ovarian cancer SK-OV-3 cells until the growth reaches the logarithmic phase, digesting with pancreatin, collecting and washing with 50mLBD tube for three times by using serum-free medium, counting, and adjusting the cell concentration to 5 × 106one/mL. Cells were inoculated subcutaneously into the right dorsal side of NOD-SCID mice, 100. mu.L of each cell suspension was inoculated. When the tumor volume is increased to 150-200 mm3At the time, mice were randomly divided into 3 groups, and a blank group, a solvent control group and a drug treatment group were set, and at least 6 mice per group were administered. The administration mode is intraperitoneal administration, and the administration period is 30 days. The body weight of the mice was recorded every 3 days during the administration period, and the presence or absence of diarrhea, convulsion, rash, significant weight loss, and the like were observed.
The in vivo tumor suppression of compound 11 is shown in figure 2 and the change in body weight of mice is shown in figure 3.
Experimental results show that when the compound 11 is administrated by the abdominal cavity at a dose of 10mg/kg per day for 30 days continuously, the growth of SK-OV-3 tumor cells in a mouse body can be effectively inhibited, and the tumor size of an administration group is obviously reduced compared with that of a blank group and a solvent control group. During administration, the weight of the mice in the administration group is not obviously reduced compared with the blank group and the solvent group, and the mice have no diarrhea, convulsion, rash and obvious toxic and side effects, which indicates that the compound has low toxicity.
Example 6K between JMJD6 protein and Compound 11DValue of
Purpose of the experiment: compound 11 was tested for its ability to bind to JMJD6 protein. Selection of biomacromolecule interaction analyzer (Biacore) for determination of K of small molecules and proteinsDThe value is obtained.
The experimental method comprises the following steps: diluting the expressed and purified JMJD6 protein to 20 mu g/mL with sodium acetate buffer solution with pH 5.5, pH 5.0, pH 4.5 and pH 4.0, performing pre-enrichment experiment using CM5 chip, selecting proper pH condition from the enrichment amount of JMJD6 on the chip surface, and adding JMJD6 under the pH condition1Proteins are covalently coupled to the chip surface. Compound 11 was subjected to a kinetic/affinity test, and six concentrations of compound 11 were formulated: 5 μ M, 2.5 μ M, 1.25 μ M, 0.625 μ M, 0.312 μ M, 0.156 μ M, binding values were determined at various concentrations following procedure autoinjection, and K for Compound 11 was obtained using fitDThe value is obtained.
K of Compound 11 with JMJD6 proteinDThe values are shown in FIG. 4.
The experimental result shows that the combination of the compound 11 and the JMJD6 protein has obvious concentration dependence, and the K of the compound isDIt was 0.755. mu.M. From the data, it is clear that compound 11 is capable of specifically binding to JMJD6 protein.
Claims (10)
1. Use of a compound of formula i or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment and/or prophylaxis of cancer:
wherein R is1、R2Independently selected from H, halogen, substituted or unsubstituted alkyl, or, R1And R2Linked to form a substituted or unsubstituted aryl group;
R3selected from substituted or unsubstituted aryl;
R4selected from H, substituted or unsubstituted alkyl, or, R4And R3Joined to form an alicyclic ring.
2. Use according to claim 1, characterized in that:
R1、R2independently selected from H, halogen, unsubstituted C1-C6 alkyl or halogen substituted C1-C6 alkyl, or R1And R2Linked to form a substituted or unsubstituted 5-to 6-membered aryl group;
preferably, R1、R2Independently selected from H, halogen, unsubstituted C1-C3 alkyl or halogen substituted C1-C3 alkyl, or R1And R2Linked to form a substituted or unsubstituted phenyl group;
preferably, R1、R2Independently selected from H or methyl, or, R1And R2Are linked to form an unsubstituted phenyl group;
preferably, R1Selected from H or methyl, R2Is H, or R1And R2Are linked to form an unsubstituted phenyl group;
further preferably, R1Is methyl, R2Is H.
3. Use according to claim 1 or 2, characterized in that:
R3selected from substituted or unsubstituted 5-to 14-membered aryl;
preferably, R3Selected from substituted or unsubstituted 5-to 6-membered aryl;
preferably, the aryl group contains 0 to 2 heteroatoms selected from nitrogen, oxygen or sulfur;
preferably, R3Selected from substituted or unsubstituted furyl, pyrrolyl, thienyl, pyrazolyl, isoxazolyl, phenyl or pyridyl;
preferably, the substituted aryl group contains at least one substituent selected from the group consisting of: hydroxy, cyano, substituted or unsubstituted alkyl, halogen, substituted or unsubstituted amino, nitro;
preferably, the substituted aryl group contains at least one substituent selected from the group consisting of: hydroxyl, cyano, unsubstituted C1-C6 alkyl, halogen substituted C1-C6 alkyl, halogen, amino, C1-C6 alkyl substituted amino and nitro;
preferably, the substituted aryl group contains at least one substituent selected from the group consisting of: hydroxyl, cyano, unsubstituted C1-C6 alkyl, halogen substituted C1-C6 alkyl, halogen, amino, C1-C6 alkyl substituted amino and nitro;
preferably, the substituted aryl group contains at least one substituent selected from the group consisting of: hydroxy, cyano, methyl, fluoro, chloro, amino, diethylamino, nitro;
preferably, R3Selected from:
4. use according to any one of claims 1 to 3, characterized in that: r4Selected from H, substituted or unsubstituted C1-C6 alkyl; preferably, R4Selected from H, unsubstituted C1-C3 alkyl or hydroxyl substituted C1-C3 alkyl; further preferably, R4Is selected from H or-CH2OH。
6. Use according to any one of claims 1 to 5, characterized in that: r4Is H.
8. use according to any one of claims 1 to 7, characterized in that: the cancer is ovarian cancer or pancreatic cancer.
9. Use according to any one of claims 1 to 8, characterized in that: the drug is a histone demethylase inhibitor drug; preferably, the drug is a histone arginine demethylase JMJD6 inhibitor drug.
10. Use according to any one of claims 1 to 9, characterized in that: the medicine is a preparation prepared by taking the compound or the pharmaceutically acceptable salt thereof as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients; preferably, the formulation is an oral formulation or an injectable formulation.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201811458195.9A CN111249283A (en) | 2018-11-30 | 2018-11-30 | Pyrimidine derivatives having anticancer effect |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201811458195.9A CN111249283A (en) | 2018-11-30 | 2018-11-30 | Pyrimidine derivatives having anticancer effect |
Publications (1)
Publication Number | Publication Date |
---|---|
CN111249283A true CN111249283A (en) | 2020-06-09 |
Family
ID=70942825
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201811458195.9A Pending CN111249283A (en) | 2018-11-30 | 2018-11-30 | Pyrimidine derivatives having anticancer effect |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN111249283A (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111808074A (en) * | 2020-07-23 | 2020-10-23 | 河南中医药大学 | Hydrazone bond-containing pyrimidinehydrazine derivative transition metal cobalt (II) complex and preparation method and application thereof |
CN113133999A (en) * | 2021-04-28 | 2021-07-20 | 广州医科大学 | Application of osthole and its derivatives in inhibiting aldehyde ketone reductase |
CN113149972A (en) * | 2021-04-09 | 2021-07-23 | 四川大学 | 2- (aryl (azacycloalkane-1-yl) methyl) phenol derivatives and use thereof |
CN116410203A (en) * | 2021-12-29 | 2023-07-11 | 四川大学 | Compound with TRK and drug-resistant mutation inhibition activity and application thereof |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005037257A2 (en) * | 2003-10-15 | 2005-04-28 | Imtm Gmbh | Novel alanyl-amino peptidase inhibitors for functionally influencing different cells and treating immunological, inflammatory, neuronal, and other diseases |
CN103570723A (en) * | 2012-07-27 | 2014-02-12 | 四川大学 | Pyrazolopyrimidine derivative, its preparation method, and its use in preparation of medicines |
WO2014083095A1 (en) * | 2012-11-29 | 2014-06-05 | Universitaet Basel | Combinations of metformin with other compounds for the treatment of cancer and for immunosuppression |
CN105017159A (en) * | 2014-04-28 | 2015-11-04 | 四川大学 | 5-fluorine-2,4-bis-substituted aminopyrimidine derivative and preparation method and application thereof |
WO2015182625A1 (en) * | 2014-05-26 | 2015-12-03 | 国立大学法人京都大学 | Ras ACTIVITY INHIBITOR AND USE THEREOF |
WO2017117430A1 (en) * | 2015-12-29 | 2017-07-06 | The Scripps Research Institute | Regulators of the endoplasmic reticulum proteostasis network |
US20180230105A1 (en) * | 2017-01-13 | 2018-08-16 | Regents Of The University Of Minnesota | Therapeutic compounds |
CN108503623A (en) * | 2018-05-11 | 2018-09-07 | 四川大学 | A kind of compound and the preparation method and application thereof inhibiting PRMT7 |
-
2018
- 2018-11-30 CN CN201811458195.9A patent/CN111249283A/en active Pending
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005037257A2 (en) * | 2003-10-15 | 2005-04-28 | Imtm Gmbh | Novel alanyl-amino peptidase inhibitors for functionally influencing different cells and treating immunological, inflammatory, neuronal, and other diseases |
CN1897928A (en) * | 2003-10-15 | 2007-01-17 | Imtm股份有限公司 | Novel alanyl-amino peptidase inhibitors for functionally influencing different cells and treating immunological, inflammatory, neuronal, and other diseases |
CN103570723A (en) * | 2012-07-27 | 2014-02-12 | 四川大学 | Pyrazolopyrimidine derivative, its preparation method, and its use in preparation of medicines |
WO2014083095A1 (en) * | 2012-11-29 | 2014-06-05 | Universitaet Basel | Combinations of metformin with other compounds for the treatment of cancer and for immunosuppression |
CN105017159A (en) * | 2014-04-28 | 2015-11-04 | 四川大学 | 5-fluorine-2,4-bis-substituted aminopyrimidine derivative and preparation method and application thereof |
WO2015182625A1 (en) * | 2014-05-26 | 2015-12-03 | 国立大学法人京都大学 | Ras ACTIVITY INHIBITOR AND USE THEREOF |
WO2017117430A1 (en) * | 2015-12-29 | 2017-07-06 | The Scripps Research Institute | Regulators of the endoplasmic reticulum proteostasis network |
US20180230105A1 (en) * | 2017-01-13 | 2018-08-16 | Regents Of The University Of Minnesota | Therapeutic compounds |
CN108503623A (en) * | 2018-05-11 | 2018-09-07 | 四川大学 | A kind of compound and the preparation method and application thereof inhibiting PRMT7 |
Non-Patent Citations (3)
Title |
---|
JING YANG等: "Jumonji domain-containing protein 6 protein and its role in cancer", 《CELL PROLIFERATION》 * |
XIANGXIAN ZHANG等: "JMJD3 in the regulation of human diseases", 《PROTEIN & CELL》 * |
XIAOAI WU等: "Discovery of KDM5A inhibitors: Homology modeling, virtual screening and structure–activity relationship analysis", 《BIOORGANIC & MEDICINAL CHEMISTRY LETTERS》 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111808074A (en) * | 2020-07-23 | 2020-10-23 | 河南中医药大学 | Hydrazone bond-containing pyrimidinehydrazine derivative transition metal cobalt (II) complex and preparation method and application thereof |
CN111808074B (en) * | 2020-07-23 | 2023-04-14 | 河南中医药大学 | Hydrazone bond-containing pyrimidinehydrazine derivative transition metal cobalt (II) complex as well as preparation method and application thereof |
CN113149972A (en) * | 2021-04-09 | 2021-07-23 | 四川大学 | 2- (aryl (azacycloalkane-1-yl) methyl) phenol derivatives and use thereof |
CN113149972B (en) * | 2021-04-09 | 2022-09-13 | 四川大学 | 2- (aryl (azacycloalkane-1-yl) methyl) phenol derivatives and use thereof |
CN113133999A (en) * | 2021-04-28 | 2021-07-20 | 广州医科大学 | Application of osthole and its derivatives in inhibiting aldehyde ketone reductase |
CN116410203A (en) * | 2021-12-29 | 2023-07-11 | 四川大学 | Compound with TRK and drug-resistant mutation inhibition activity and application thereof |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN111249283A (en) | Pyrimidine derivatives having anticancer effect | |
Hannoun et al. | Synthesis and antibacterial evaluation of a novel library of 2-(thiazol-5-yl)-1, 3, 4-oxadiazole derivatives against methicillin-resistant Staphylococcus aureus (MRSA) | |
US9255107B2 (en) | Heteroaryl alkyne compound and use thereof | |
CN108578412A (en) | A kind of taxol and diaza * and carbazole compound drug combination compositions | |
CN107445896B (en) | Phenyl hydroxamic acid compound with anti-tumor activity and application thereof | |
EP4116302A1 (en) | Compound and preparation method thereof and application thereof in preparation of therapeutic anti-cancer drug | |
EP2826780B1 (en) | Thieno[2,3-d]pyridazine derivatives and therapeutic use thereof for protein kinase inhibition | |
Zhan et al. | Design, synthesis, and biological evaluation of novel highly selective polo-like kinase 2 inhibitors based on the tetrahydropteridin chemical scaffold | |
US20110166191A1 (en) | 3-(2-amino-ethyl)-5-(3-cyclohexyl-propylidene)-thiazolidine-2,4-dione and its derivatives as multiple signaling pathway inhibitors and for the treatment of cancer | |
Ragab et al. | Design, synthesis, molecular dynamic simulation studies, and antibacterial evaluation of new spirocyclic aminopyrimidines | |
CN101058573B (en) | 2-hydrazone substituted triazine compound, preparation method thereof, and medicinal composition and use using same as active component | |
CN108752412B (en) | Boswellic acid derivatives and their use | |
EP3805193A1 (en) | Novel biphenyl derivative compound and use thereof | |
CN109293588B (en) | Small molecular compound with IDO1/TDO dual-target, and preparation method and application thereof | |
CN111662271B (en) | Compound with IDH mutant inhibitory activity and preparation method and application thereof | |
CN109438279B (en) | Small molecule compound for overcoming EGFR drug-resistant mutation and preparation method and application thereof | |
CN108239081B (en) | Compound for inhibiting ROCK and application thereof | |
CN112250675B (en) | Tyrosine kinase inhibitor containing amide structure and preparation method thereof | |
CN116239594B (en) | 6- (imidazo [1,2-a ] pyridin-6-yl) quinazoline derivatives and uses thereof | |
CN108210508A (en) | A kind of antineoplastic pharmaceutical compositions | |
CN112300138B (en) | Tyrosine kinase inhibitor containing ester group structure and preparation method thereof | |
CN114105958B (en) | Hydroximic acid-containing 2-phenylamino pyrimidine derivatives and application thereof | |
CN108658874B (en) | Thiopyrimidine heterocyclic antitumor compound and preparation method and application thereof | |
CN108245515A (en) | The synthesis and its application of multi-substituted isoquinoline derivative | |
CN102688232B (en) | Synthesis and application of isoindole-1, 3-dione derivative as RSK2 inhibitor |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200609 |