CN114920859B - 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative and preparation method and application thereof - Google Patents
5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative and preparation method and application thereof Download PDFInfo
- Publication number
- CN114920859B CN114920859B CN202210511426.8A CN202210511426A CN114920859B CN 114920859 B CN114920859 B CN 114920859B CN 202210511426 A CN202210511426 A CN 202210511426A CN 114920859 B CN114920859 B CN 114920859B
- Authority
- CN
- China
- Prior art keywords
- fluorouracil
- cyclodextrin
- butyryl
- beta
- derivative
- 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.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
- C08B37/0012—Cyclodextrin [CD], e.g. cycle with 6 units (alpha), with 7 units (beta) and with 8 units (gamma), large-ring cyclodextrin or cycloamylose with 9 units or more; Derivatives thereof
-
- 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
- A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/716—Glucans
- A61K31/724—Cyclodextrins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The invention provides a 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative, a preparation method and application thereof, and relates to the technical field of organic synthesis, wherein 5-fluorouracil-1-butyric acid, N-carbonyl diimidazole and cyclodextrin are used as raw materials to prepare the 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative, the reaction process steps are simple to operate, and the reaction conditions are mild; the synthesized 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative is used as a prodrug of 5-fluorouracil, can improve the in-vivo dynamic process of the drug and increase the stability of the drug; after the second side derivative of 5-fluorouracil-1-butyryl-beta-cyclodextrin is taken by a patient with colon cancer, the second side derivative of 5-fluorouracil-1-butyryl-beta-cyclodextrin can always increase tumor cells, and the toxic and side effects on human bodies are weakened.
Description
Technical Field
The invention belongs to the technical field of organic synthesis, and particularly relates to a 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative, a preparation method and application thereof.
Background
5-Fluorouracil (5-Fluorouracil, 5-FU) with chemical formula C 4 H 3 FN 2 O 2 Also known as fluorouracil, is a pyrimidine analog which is a white or off-white crystalline or crystalline powder, slightly soluble in water, slightly soluble in ethanol, and almost insoluble in chloroform; dissolved in dilute hydrochloric acid or sodium hydroxide solution. 5-fluorouracil as pyrimidine fluoride, which is an antimetaboliteAntitumor agents inhibit thymine nucleotide synthase, block the conversion of deoxypyrimidine nucleotides to thymidine nuclei, and interfere with DNA synthesis. Has a certain inhibition effect on RNA synthesis. The action mechanism of the 5-FU is as follows: inhibition of deoxythymidylate synthase by its conversion to 5-fluorouracil deoxynucleotide (5F-dUMP) in vivo, prevents the conversion of deoxyuridylate (dUMP) methyl to deoxythymidylate (dTMP), and thus affects DNA synthesis; on the other hand, due to the enhancement of the stable C-F bond structure and acidity of the 5-FU, the 5-FU can be more firmly combined with enzyme, and after being ingested into cells, uracil which is an important precursor of tumor nucleic acid is replaced at the molecular level, biological macromolecules are fraudulently doped, abnormal RNA is formed, the function of the nucleic acid is affected, and the gene mutation is caused. That is, when 5-FU enters the body, it inhibits the synthesis of DNA and RNA of tumor cells, resulting in death of tumor cells in the proliferation stage.
In the prior art, the 5-FU is clinically used for treating digestive tract tumors, breast cancer, ovarian cancer, chorionic epithelial cancer, cervical cancer, liver cancer, bladder cancer, skin cancer (local smearing) and leukoplakia vulvae (local smearing) and the like. However, long-term administration of 5-FU can cause great toxic and side effects to patients, such as short half-life in vivo, nausea, emesis and other adverse gastrointestinal reactions.
Disclosure of Invention
In view of the above, the present invention provides a second side derivative of 5-fluorouracil-1-butyryl-beta-cyclodextrin to solve the technical problem that long-term administration of 5-FU in the prior art causes a large toxic and side effect to patients.
It is also necessary to provide a process for the preparation of the second side derivatives of 5-fluorouracil-1-butyryl-beta-cyclodextrin.
It is also desirable to provide the use of a second side derivative of 5-fluorouracil-1-butyryl-beta-cyclodextrin.
The technical scheme adopted for solving the technical problems is as follows:
a 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative, which has a chemical structural formula:
n＝1-7。
preferably, 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative is prepared by taking 5-fluorouracil-1-butyric acid, N-carbonyl diimidazole and cyclodextrin as raw materials, and comprises the following steps:
s1: adding an organic solvent A and N, N-carbonyl diimidazole into 5-fluorouracil-1-butyric acid, and reacting for a first preset time at room temperature to obtain a 5-fluorouracil-1-butyric acid imidazole amide mixed solution;
s2: adding cyclodextrin and a catalyst B into the 5-fluorouracil-1-butyric acid imidazole amide mixed solution prepared in the step S2, and reacting for a second preset time at room temperature to obtain a mixed solution of 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative;
s3: performing rotary evaporation on the mixed solution of the 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative, and performing chromatography on the product after rotary evaporation to obtain a second surface derivative liquid of a double-surface monosubstituted compound 5-fluorouracil-1-butyryl-beta-cyclodextrin;
s4: and (3) performing liquid rotary evaporation on the second side derivative of the dihedral monosubstituted compound 5-fluorouracil-1-butyryl-beta-cyclodextrin to obtain a solid of the second side derivative of the 5-fluorouracil-1-butyryl-beta-cyclodextrin.
Preferably, in the step S1, the organic solvent a used is N, N-Dimethylformamide (DMF).
Preferably, in the step S1, the first predetermined time is 1-2h.
Preferably, in the step S2, the catalyst B used is triethylamine (Et 3 N)。
Preferably, in the step S2, the second predetermined time is 18-24 hours.
Preferably, in the S1, the molar ratio of the 5-fluorouracil-1-butyric acid to the N, N-carbonyldiimidazole is 1: (1-2).
Preferably, the DMF is anhydrous DMF.
Preferably, in the S2, the molar ratio of the 5-fluorouracil-1-butyric acid imidazole amide to cyclodextrin and triethylamine is 1: (1-1.5): (5-10).
Use of a 5-fluorouracil-1-butyryl-beta-cyclodextrin second-side derivative as described above, in a medicament for the treatment of colon cancer.
Compared with the prior art, the invention has the beneficial effects that:
the invention takes 5-fluorouracil-1-butyric acid, N-carbonyl diimidazole and cyclodextrin as raw materials to prepare the 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative, and the reaction process has simple operation steps and mild reaction conditions; the synthesized 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative is used as a prodrug of 5-fluorouracil, and the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative has better slow release and long-acting functions, can improve the in-vivo dynamic process of a medicament and increase the stability of the medicament; and after the colon cancer patient takes the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative, the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative can inhibit the growth of tumor cells, and the toxic and side effects on human bodies are reduced.
Drawings
FIG. 1 is a nuclear magnetic resonance hydrogen spectrum of ethyl 5-fluorouracil-1-butyrate in example one.
FIG. 2 is a nuclear magnetic resonance carbon spectrum of ethyl 5-fluorouracil-1-butyrate in example one.
FIG. 3 is a mass spectrum of ethyl 5-fluorouracil-1-butyrate in example one.
FIG. 4 is a nuclear magnetic resonance hydrogen spectrum of 5-fluorouracil-1-butyric acid in example one.
FIG. 5 is a nuclear magnetic resonance carbon spectrum of 5-fluorouracil-1-butyric acid in example one.
FIG. 6 is a mass spectrum of 5-fluorouracil-1-butyric acid in example one.
FIG. 7 is a nuclear magnetic resonance hydrogen spectrum of 5-fluorouracil-1-butyryl-beta-cyclodextrin in example one.
FIG. 8 is a nuclear magnetic resonance carbon spectrum of 5-fluorouracil-1-butyryl-beta-cyclodextrin in example one.
FIG. 9 is a mass spectrum of 5-fluorouracil-1-butyryl-beta-cyclodextrin in example one.
FIG. 10 is a graph of data analysis of inhibition of HT-29 colon cancer cell proliferation by 5-fluorouracil-1-butyryl-beta cyclodextrin in example one.
FIG. 11 is a graph of data analysis of inhibition of NCM-460 normal colon epithelial cell proliferation by 5-fluorouracil-1-butyryl-beta cyclodextrin in example one.
FIG. 12 is a graph showing the growth of nude mice tumors after intervention with different compounds according to example one.
Fig. 13 shows the change in body weight of nude mice after intervention with different compounds in example one.
Detailed Description
The technical scheme and technical effects of the embodiments of the present invention are further described in detail below with reference to the accompanying drawings.
A 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative, which has a chemical structural formula:
n＝1-7。
compared with the prior art, the invention has the beneficial effects that:
the invention takes 5-fluorouracil-1-butyric acid, N-carbonyl diimidazole and cyclodextrin as raw materials to prepare the 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative, and the reaction process has simple operation steps and mild reaction conditions; the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative is selectively generated through a system formed by 5-fluorouracil-1-butyric acid, N-carbonyl diimidazole, triethylamine and DMF, and the synthesized 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative is used as a prodrug of 5-fluorouracil, and the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative has better slow release and long-acting functions, can improve the in-vivo dynamics process of a medicament and increase the stability of the medicament; and after the colon cancer patient takes the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative, the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative can inhibit the growth of tumor cells, and the toxic and side effects on human bodies are reduced.
Specifically, the 5-fluorouracil-1-butyric acid is prepared by the following method: adding DMF into 5-fluorouracil, measuring a proper amount of triethylamine, adding into a reaction bottle, magnetically stirring for 30 minutes, slowly adding ethyl 4-bromobutyrate into the reaction liquid, reacting for 6-8 hours to obtain a mixed solution, adding a proper amount of distilled water into the mixed solution, quenching the mixed solution, reacting, extracting with dichloromethane for 3 times, washing the extract with distilled water for 3 times, purifying by silica gel column chromatography to obtain ethyl 5-fluorouracil-1-butyrate, adding water and methanol solution into the ethyl 5-fluorouracil-1-butyrate, adding NaOH, reacting for 2 hours, regulating the pH of the reaction liquid to 2, finding that solid is not precipitated, extracting the residual reaction liquid with ethyl acetate for 4-5 times after removing methanol by rotary evaporation, and spin-drying an ethyl acetate layer to obtain the target product 5-fluorouracil-1-butyric acid.
Specific: the molar ratio of 5-fluorouracil to ethyl 4-bromobutyrate is 1: (1-1.5).
Further, 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative is prepared by taking 5-fluorouracil-1-butyric acid, N-carbonyl diimidazole and cyclodextrin as raw materials, and comprises the following steps:
s1: adding an organic solvent A and N, N-carbonyl diimidazole into 5-fluorouracil-1-butyric acid, and reacting for a first preset time at room temperature to obtain a 5-fluorouracil-1-butyric acid imidazole amide mixed solution;
s2: adding cyclodextrin and a catalyst B into the 5-fluorouracil-1-butyric acid imidazole amide mixed solution prepared in the step S2, and reacting for a second preset time at room temperature to obtain a mixed solution of 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative;
s3: performing rotary evaporation on the mixed solution of the 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative, and performing chromatography on the product after rotary evaporation to obtain a second surface derivative liquid of a double-surface monosubstituted compound 5-fluorouracil-1-butyryl-beta-cyclodextrin;
s4: and (3) performing liquid rotary evaporation on the second side derivative of the dihedral monosubstituted compound 5-fluorouracil-1-butyryl-beta-cyclodextrin to obtain a solid of the second side derivative of the 5-fluorouracil-1-butyryl-beta-cyclodextrin.
Further, in the step S1, the organic solvent a is N, N-Dimethylformamide (DMF), and DMF not only plays a role of a solvent, but also plays a role of a catalyst, so that the reaction between 5-fluorouracil-1-butyric acid and N, N-carbonyl diimidazole is quicker and more severe, and the amount of DMF does not change during the reaction.
Further, in the step S1, the first predetermined time is 1-2h.
Further, in the step S2, the catalyst B used is triethylamine (Et 3 N)。
Specifically, the second side derivative of 5-fluorouracil-1-butyryl-beta-cyclodextrin is selectively generated through a system formed by 5-fluorouracil-1-butyric acid, N-carbonyl diimidazole, triethylamine and DMF.
Further, in the step S2, the second predetermined time is 18-24 hours.
Further, in the S1, a molar ratio of the 5-fluorouracil-1-butyric acid to the N, N-carbonyldiimidazole is 1: (1-2).
Further, the DMF is anhydrous super-dry DMF.
Further, in the S2, the molar ratio of the 5-fluorouracil-1-butyric acid imidazole amide to cyclodextrin and triethylamine is 1: (1-1.5): (5-10).
Use of a 5-fluorouracil-1-butyryl-beta-cyclodextrin second-side derivative as described above, in a medicament for the treatment of colon cancer.
For ease of understanding, the invention is further illustrated by the following examples:
embodiment one:
6.5g (50 mmol) of 5-fluorouracil is added into a 250mL single-neck flask, 100mL of ultra-dry DMF is added to dissolve the 5-fluorouracil, 4.17mL of triethylamine is added as a catalyst, the mixture is stirred by a magnetic stirrer, the reaction solution is fully mixed for 30 minutes, 8.59mL (60 mmol) of ethyl 4-bromobutyrate is slowly added into a reaction bottle, the reaction is continued for 6 to 8 hours at room temperature after the dripping is finished, a proper amount of distilled water is added into the reaction solution to quench the reaction after the reaction is finished, the reaction solution is extracted for 3 times by dichloromethane, the extract is washed for 3 times by water, and the extract is purified by silica gel column chromatography. Obtaining 5-fluorouracil-1-ethyl butyrate; dissolving 5-fluorouracil-1-ethyl butyrate in a mixed solution of 14mL of water and 28mL of methanol, adding 0.983g (24.57 mmol) of NaOH, starting stirring, adjusting the pH of the reaction solution to 2 after 2 hours, extracting the residual reaction solution with ethyl acetate for 4-5 times after removing methanol by rotary evaporation, obtaining a product 5-fluorouracil-1-butyric acid after spin drying an ethyl acetate layer, accurately weighing 1.2g (5.56 mmol), placing into a 250mL reaction bottle, adding 150mL of LDMF to dissolve the product, adding 1.081g (6.67 mmol) of N, N-carbonyl diimidazole into the reaction bottle, starting stirring, reacting for 1-2 hours at room temperature to obtain 5-fluorouracil-1-butyric acid imidazole amide, adding 6.31g (5.56 mmol) of beta-cyclodextrin (beta-CD) and 6.18mL of catalyst triethylamine, and stirring at room temperature to react for 18 h to obtain a second mixed solution of 5-fluorouracil-1-butyryl imidazole; and (3) performing rotary evaporation on the obtained mixed solution of the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative by a rotary evaporator to obtain about 5mL of DMF, adding acetone, stirring, precipitating a reaction product, performing suction filtration to obtain a white solid, performing C18 reversed phase column chromatography on the white solid, refining to obtain a liquid of the second side derivative of the dihomo-substituted compound 5-fluorouracil-1-butyryl-beta-cyclodextrin, and performing rotary evaporation on the liquid by the rotary evaporator to obtain a solid of the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative (5 FuBA-beta CD), wherein the substitution degree is 1.
Referring to fig. 1, 2 and 3, the ethyl 5-fluorouracil-1-butyrate prepared in the above examples was subjected to nmr and mass spectrometry, respectively; wherein the method comprises the steps of 1 H NMR spectrum (FIG. 1), 13 C NMR spectrum (fig. 2), mass spectrum (fig. 3):
please refer to fig. 1 and 2, it can be seen that: 1 H NMR(400MHz，DMSO-d6)δ11.75(d，1H，J＝5.2Hz，N 3 -H)，8.06(d，1H，J＝6.9Hz)，4.02(q，2H，J＝7.1Hz，-OCH 2 CH 3 )，3.64(t，2H，J＝6.8Hz，N-CH 2 -CH 2 -CH 2 )，2.33(t，2H，J＝14.7Hz，-CH 2 CH 2 COO)，1.84(m，2H，N-CH 2 -CH 2 -CH 2 )，1.16(t，3H，J＝7.1Hz，-COOCH 2 CH 3 )。
13 C NMR(100MHz，DMSO-d6)δ172.74，158.05，157.79，150.11，141.21，138.94，130.67，130.34，60.37，47.51，30.85，23.96，14.51。
mass spectrometry data: ESI-MS (m/z): 243.2[ M-H ] + ].
As can be seen from the nuclear magnetic hydrogen spectrum data of FIG. 1, the chemical shift value 11.75 is the hydrogen atom on the pyrimidine ring N3, and the chemical shift value is 8.06 is the hydrogen atom on the 6-position of the pyrimidine ring. The chemical shift value of the hydrogen atom at the N1 position on the pyrimidine ring of 5-fluorouracil is 10.73, and no hydrogen atom with the chemical shift value of about 10.73 is found in the nuclear magnetic resonance hydrogen spectrum of ethyl 5-fluorouracil-1-butyrate, which indicates that the hydrogen atom at the N1 position is completely replaced by ethyl butyrate. Further, the ion peak of 243.2 was found to be a signal peak of ethyl 5-fluorouracil-1-butyrate in negative ion mode by detection by electrospray mass spectrometry. The above results all confirm that ethyl 4-bromobutyrate is linked to the N1 position of 5-fluorouracil, forming the intermediate ethyl 5-fluorouracil-1-butyrate.
Performing nuclear magnetic resonance detection on the 5-fluorouracil-1-butyric acid prepared in the previous example; wherein the method comprises the steps of 1 H NMR spectrum (FIG. 4), 13 C NMR spectrum (fig. 5), mass spectrum (fig. 6).
Please refer to fig. 5 and 6, it can be seen that: 1 H NMR(400MHz，DMSO-d 6 )δ11.73(d，1H，J＝5.2Hz，N 3 -H)，10.51(s，1H，-CH 2 -CH 2 -CH 2 -COOH)，8.06(d，1H，J＝6.8Hz)，3.64(t，2H，J＝6.9Hz，N-CH 2 -CH 2 -CH 2 )，2.25(t，2H，J＝7.4Hz，-CH 2 CH 2 COO)，1.81(q，2H，J＝7.2Hz，N-CH 2 -CH 2 -CH 2 )。
13 C NMR(101MHz，DMSO-d 6 )δ174.28，158.07，157.81，150.11，141.18，138.91，130.74，130.41，47.59，31.01，24.08。
mass spectrometry data: ESI-MS (m/z): 215.1[ M-H ] + ].
As can be seen from the nuclear magnetic resonance hydrogen spectrum data of FIG. 4, the chemical shift values 4.02 and 1.16 are the hydrogen signals disappeared. Further, it was found that the ion peak of 215.1 was a signal peak of 5-fluorouracil-1-butyric acid in the negative ion mode by electrospray detection, and the above results confirmed that ethyl 5-fluorouracil-1-butyrate was hydrolyzed to 5-fluorouracil-1-butyric acid.
Referring to fig. 7, 8 and 9, the second side derivatives of 5-fluorouracil-1-butyryl-beta-cyclodextrin prepared in the above examples were subjected to nmr and mass spectrometry, respectively; wherein the method comprises the steps of 1 H NMR spectrum (FIG. 7), 13 C NMR spectrum (fig. 8), mass spectrum (fig. 9):
please refer to fig. 7 and 8, it can be seen that: 1 H NMR(400MHz，DMSO-d 6 )δ8.01(dd，1H，J＝11.8Hz，6.5Hz)，5.88(s，1H)，5.81(s，2H)，5.74(d，6H，J＝22.5Hz)，4.83(s，7H)，4.65(s，1H)，4.52(t，6H，J＝9.1Hz)，4.45-4.34(m，2H)，3.85(d，1H，J＝9.8Hz)，3.73-3.59(m，20H)，3.38-3.25(m，11H)，2.41-2.30(m，1H)，1.83(m，2H)。
13 C NMR(101MHz，DMSO-d 6 )δ172.93，172.5，150.70，150.45，130.47，130.14，102.37-101.75(C-1)，98.78(C-1’)，82.16-81.01(C-4)，78.65(C-4’)，73.49，72.48，69.72，60.35，47.65，31.22，30.71，30.11，23.97，21.13。
fig. 9, mass spectrometry data: ESI-MS (m/z): 243.2[ M-H ] + ].
From the examination of FIGS. 7 to 9, it was found by the analysis of the nuclear magnetic resonance hydrogen spectrum that the chemical shift value of the 6-position hydrogen atom on the pyrimidine ring was 8.01, the chemical shift value of the 1-position hydrogen atom on the beta-cyclodextrin was 4.83, and the number ratio of the 6-position hydrogen atom on the pyrimidine ring to the 1-position hydrogen atom on the beta-cyclodextrin was 1:7, so that it was confirmed that the 5-fluorouracil derivative was monosubstituted with the beta-cyclodextrin. The chemical shift of the carbon atom at the 1-position of the beta-cyclodextrin is found to be respectively shifted from 102.37 to 98.78 by nuclear magnetic resonance carbon spectrum analysis. Meanwhile, the chemical shift value of the carbon atom at the 4-position of the beta-cyclodextrin is respectively shifted from 82.16 to 78.65, which indicates that the 5-fluorouracil derivative is connected with the secondary hydroxyl of the second surface of the beta-cyclodextrin through an ester bond. In addition, the ion peak of 1332.8 was found to be the signal peak of 5-fluorouracil-1-butyryl-beta-cyclodextrin in the negative ion mode by mass spectrometry detection, and it was further confirmed that the 5-fluorouracil derivative and beta-cyclodextrin were monosubstituted, and the above result confirmed that the second side derivative of 5-fluorouracil-1-butyryl-beta-cyclodextrin was synthesized.
It was thus possible to determine that the solid 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative of the final product prepared was consistent with the final product in the pre-established synthetic route.
The following experiment was performed on the prepared 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative.
1. Antitumor Activity assay
The inhibition of proliferation of colon cancer cells was detected by CCK-8 method using 5-fluorouracil-1-butyryl-beta cyclodextrin (5 FUBA-beta CD). Taking logarithmic growth cycle, digesting and blowing human colon cancer HT-29 cells with good growth state into single cell suspension, inoculating 5×103 cells per well into 96-well plate, placing at 37deg.C, 5% CO 2 Culturing for 24h. Cells were subjected to a synchronization treatment by serum starvation, a blank group (without cells and drugs), a control group (without drugs) and a dosing experimental group were set, 4 compound wells each, 200 μl of drug-containing medium was added in the concentration order of 0, 5, 10, 20, 40, 80, 160 μΜ, and then 96 well plates were placed in a cell incubator for 24, 48, 72 hours, respectively. Finally, according to the cell proliferation and toxicity detection kit (CCK-8), the old culture medium is sucked and removed, 100 μl of serum-free culture medium containing 10% CCK-8 solution is added to each well, the mixture is incubated at 37deg.C for 0.5h, and the absorbance (450 nm) of each well is measured by an enzyme-labeled instrument and repeated 3 times. Cell viability for each group of cell proliferation was calculated according to the following formula, cell viability (%) = (OD value of dosing experimental group-OD value of blank group)/(OD value of control group-OD value of blank group) ×100%, and experimental results are shown in fig. 10.
As can be seen from FIG. 10 (inhibition rate of HT-29 colon cancer cell proliferation by 5-fluorouracil-1-butyryl-beta cyclodextrin), the in vitro antitumor activity of the synthesized 5-fluorouracil-1-butyryl-beta cyclodextrin was evaluated by CCK-8 method. Cell viability at different concentrations and at different time points of action was studied with 5Fu as positive control. The experimental results show that the compound has concentration dependence and time dependence on the inhibition of colorectal cancer HT-29 cell proliferation, and the 5-fluorouracil-1-butyryl-beta cyclodextrin has not shown good anti-tumor activity, and can be an in vitro cell culture environment unfavorable for release and in vitro lack of enzymes required for metabolism.
2. Cytotoxicity test
Toxicity of 5-fluorouracil-1-butyryl-beta cyclodextrin to normal human colon epithelial cells was detected by CCK-8 method. Taking logarithmic growth cycle, digesting and blowing normal human colon epithelial NCM-460 cells with good growth state into single cell suspension, inoculating 1×104 cells per well into a 96-well plate, and culturing at 37 ℃ for 24h in 5% CO 2. Cells were subjected to a synchronization treatment by serum starvation, a blank group (without cells and drugs), a control group (without drugs) and a dosing experimental group were set, 4 compound wells each, 200 μl of drug-containing medium was added in the concentration order of 0, 5, 10, 20, 40, 80, 160 μΜ, and then 96 well plates were placed in a cell incubator for 24, 48, 72 hours, respectively. Finally, according to the cell proliferation and toxicity detection kit (CCK-8), the old culture medium is sucked and removed, 100 μl of serum-free culture medium containing 10% CCK-8 solution is added to each well, the mixture is incubated at 37deg.C for 1.5h, and the absorbance (450 nm) of each well is measured by an enzyme-labeled instrument and repeated 3 times. Calculation of cell proliferation activity referring to the above formula, the experimental results are shown in fig. 11.
As can be seen from FIG. 11 (inhibition rate of NCM-460 normal colon epithelial cell proliferation by 5-fluorouracil-1-butyryl-beta cyclodextrin), we tested the toxicity of 5-fluorouracil-1-butyryl-beta cyclodextrin on human normal colon epithelial NCM-460 cells at different concentrations and for different times of action by CCK-8 method. Compared with the positive control 5Fu, the inhibition of 5-fluorouracil-1-butyryl-beta cyclodextrin on cells does not have obvious increase along with the increase of concentration and the extension of action time, which can indicate that the 5-fluorouracil-1-butyryl-beta cyclodextrin has weaker toxicity on normal colon cells, and therefore, the 5-fluorouracil-1-butyryl-beta cyclodextrin still has certain specificity and selectivity on tumor cells.
3. Establishment of nude mouse humanized colon cancer model and administration scheme
Amplifying and culturing HT-29 humanized colon cancer cells with good growth state in logarithmic growth cycle, re-suspending with serum-free culture medium to obtain cell suspension, and regulating cell count to 3×10 7 After local skin disinfection of the nude mice with 75% alcohol per ml, 100ul of cell suspension was injected into the right armpit of each nude mouse with a 1ml sterile syringe. After 10 days of inoculation, the nude mice have good growth state of subcutaneous tumor, the tumor forming rate is 100 percent, and the average tumor volume of the nude mice is more than 100mm 3 The patients were divided into 4 groups according to a random number method, 8 groups were administered with corresponding drug solutions by intraperitoneal injection every other day, and continuous observation was carried out for 24 days, wherein the model control group was administered with an equal volume of physiological saline, and the administration schedule is shown in table 1.
TABLE 1
And (5) observing body surface indexes of the nude mice.
The weight of the nude mice is measured before the administration every other day in the administration period, and a weight change curve is drawn after recording; the tumor volume of the nude mice is measured by a vernier caliper, a tumor growth curve is drawn, the nude mice are sacrificed after the end of the administration, spleens of the nude mice are weighed, spleen indexes are calculated according to a formula, the growth curve of the tumor of the nude mice is obtained after different compounds are interfered, and the body weight of the nude mice is changed after the different compounds are interfered, as shown in fig. 12.
Fig. 12 (growth curve of nude mice tumor after intervention of different compounds) shows the increasing trend of tumor volume of each group of nude mice, and it can be judged from the tumor growth curve that the tumor growth speed is the fastest, and the tumor growth speed of each other experimental group is relatively slow. There was no significant difference in 5-fluorouracil compared to 5 Fu. Therefore, the 5-fluorouracil-butyryl-beta cyclodextrin second side derivative plays a certain role in tumor inhibition effect.
Fig. 13 (weight change of nude mice after intervention of different compounds) shows that the weight of each group of nude mice after administration has a small-amplitude increasing trend in the initial period of administration, the weight of the nude mice in the model group and the 5 FuBA-beta CD group has obvious reduction along with the extension of the administration days, and death occurs on the 18 th day and the 20 th day, so that the effect on the weight of the nude mice is the greatest, and the 5-fluorouracil-butyryl-beta cyclodextrin has a certain attenuation effect.
The chemical equation of the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative prepared by the invention is as follows:
the foregoing disclosure is illustrative of the preferred embodiments of the present invention, and is not to be construed as limiting the scope of the invention, as it is understood by those skilled in the art that all or part of the above-described embodiments may be practiced with equivalents thereof, which fall within the scope of the invention as defined by the appended claims.
Claims (5)
1. A 5-fluorouracil-1-butyryl- β -cyclodextrin second side derivative, characterized by: the chemical structural formula is as follows:
n＝1-7；
taking 5-fluorouracil-1-butyric acid, N-carbonyl diimidazole and cyclodextrin as raw materials to prepare a 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative, which comprises the following steps:
s1: adding an organic solvent A and N, N-carbonyl diimidazole into 5-fluorouracil-1-butyric acid, and reacting for a first preset time at room temperature to obtain a 5-fluorouracil-1-butyric acid imidazole amide mixed solution;
s2: adding cyclodextrin and a catalyst B into the 5-fluorouracil-1-butyric acid imidazole amide mixed solution prepared in the step S1, and reacting for a second preset time at room temperature to obtain a mixed solution of 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative;
s3: performing rotary evaporation on the mixed solution of the 5-fluorouracil-1-butyryl-beta-cyclodextrin second surface derivative, and performing chromatography on the product after rotary evaporation to obtain a second surface derivative liquid of a double-surface monosubstituted compound 5-fluorouracil-1-butyryl-beta-cyclodextrin;
s4: performing liquid rotary evaporation on a second side derivative of a second side monosubstituted compound 5-fluorouracil-1-butyryl-beta-cyclodextrin to prepare a second side derivative solid of 5-fluorouracil-1-butyryl-beta-cyclodextrin, wherein the second side derivative of 5-fluorouracil-1-butyryl-beta-cyclodextrin has low toxicity to normal colon epithelial cells;
in the step S1, the organic solvent A is N, N-Dimethylformamide (DMF), and the first preset time is 1-2h;
in the step S2, the catalyst B used is triethylamine (Et 3 N), the second predetermined time is 18-24h.
2. The method for preparing a second side derivative of 5-fluorouracil-1-butyryl- β -cyclodextrin according to claim 1, wherein in S1, the molar ratio of 5-fluorouracil-1-butyric acid to N, N-carbonyldiimidazole is 1: (1-2).
3. The method of preparing a second side derivative of 5-fluorouracil-1-butyryl- β -cyclodextrin according to claim 1 wherein the DMF is anhydrous DMF.
4. The method for preparing a second side derivative of 5-fluorouracil-1-butyryl-beta-cyclodextrin according to claim 1, wherein in S2, the molar ratio of 5-fluorouracil-1-butyryl-beta-cyclodextrin imidazole amide to cyclodextrin and triethylamine is 1: (1-1.5): (5-10).
5. Use of a second side derivative of 5-fluorouracil-1-butyryl- β -cyclodextrin according to claim 1 or 2, characterized in that: the application of the 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative in preparing a medicament for treating colon cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210511426.8A CN114920859B (en) | 2022-05-11 | 2022-05-11 | 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210511426.8A CN114920859B (en) | 2022-05-11 | 2022-05-11 | 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114920859A CN114920859A (en) | 2022-08-19 |
| CN114920859B true CN114920859B (en) | 2023-09-26 |
Family
ID=82808599
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210511426.8A Active CN114920859B (en) | 2022-05-11 | 2022-05-11 | 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114920859B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5631244A (en) * | 1993-05-13 | 1997-05-20 | L'oreal | mono(6-amino-6-deoxy)cyclodextrin derivatives substituted in the 6-position by an α-amino acid residue, process for their preparation and their uses |
| CN101657203A (en) * | 2007-03-02 | 2010-02-24 | 卧龙岗大学 | Compositions and methods for delivery of anti-cancer agents |
| EP2422818A2 (en) * | 2010-08-27 | 2012-02-29 | Tecnimede-Sociedade Tecnico-Medicinal, S.A. | Process to produce a diclofenac cyclodextrin conjugate |
| CN102525933A (en) * | 2010-12-27 | 2012-07-04 | 上海高科联合生物技术研发有限公司 | Poly-L-lactic acid and polyethyleneglycol block copolymer (PLLA-PEG) supported 5-fluorouracil nanoparticles and preparation method thereof |
| CN108976318A (en) * | 2017-06-01 | 2018-12-11 | 首都医科大学 | Mono- 6-(biotin amido group) -6- deoxidation-beta-cyclodextrin and its preparation method and application |
| CN112940152A (en) * | 2021-01-31 | 2021-06-11 | 宁夏医科大学 | 5-fluorouracil-1-methoxycarbonyl-type cyclodextrin derivative and preparation method thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8057821B2 (en) * | 2004-11-03 | 2011-11-15 | Egen, Inc. | Biodegradable cross-linked cationic multi-block copolymers for gene delivery and methods of making thereof |
-
2022
- 2022-05-11 CN CN202210511426.8A patent/CN114920859B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5631244A (en) * | 1993-05-13 | 1997-05-20 | L'oreal | mono(6-amino-6-deoxy)cyclodextrin derivatives substituted in the 6-position by an α-amino acid residue, process for their preparation and their uses |
| CN101657203A (en) * | 2007-03-02 | 2010-02-24 | 卧龙岗大学 | Compositions and methods for delivery of anti-cancer agents |
| EP2422818A2 (en) * | 2010-08-27 | 2012-02-29 | Tecnimede-Sociedade Tecnico-Medicinal, S.A. | Process to produce a diclofenac cyclodextrin conjugate |
| CN102525933A (en) * | 2010-12-27 | 2012-07-04 | 上海高科联合生物技术研发有限公司 | Poly-L-lactic acid and polyethyleneglycol block copolymer (PLLA-PEG) supported 5-fluorouracil nanoparticles and preparation method thereof |
| CN108976318A (en) * | 2017-06-01 | 2018-12-11 | 首都医科大学 | Mono- 6-(biotin amido group) -6- deoxidation-beta-cyclodextrin and its preparation method and application |
| CN112940152A (en) * | 2021-01-31 | 2021-06-11 | 宁夏医科大学 | 5-fluorouracil-1-methoxycarbonyl-type cyclodextrin derivative and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| Regioselective synthesis, stability and release behaviors of the secondary 5-fluorouracil acetic acid/b-cyclodextrin conjugate for colon delivery;Shi Jie Wei等;《J Incl Phenom Macrocycl Chem》;20151207;第84卷;第43-47页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114920859A (en) | 2022-08-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111559991B (en) | Preparation method and application of naphthylamine compound and salt thereof | |
| CN114315929B (en) | Geniposide derivative with xanthine oxidase inhibitory activity, and preparation method and application thereof | |
| CN113444035B (en) | Naphthyl urea compounds with anticancer effect and preparation method and application thereof | |
| CN112979528B (en) | Tegaserod water-soluble organic acid salt and preparation method and application thereof | |
| CN114907274A (en) | 5-fluorouracil-1-alkyl acid derivative, preparation method and application thereof | |
| CN114920859B (en) | 5-fluorouracil-1-butyryl-beta-cyclodextrin second side derivative and preparation method and application thereof | |
| CN110294764B (en) | Azo bond-connected podophyllotoxin derivative and preparation method thereof | |
| CN114671751B (en) | O-hydroxyphenylketone compound and preparation method and application thereof | |
| CN111518546A (en) | Hypoxic microenvironment response fluorescent probe and preparation method and application thereof | |
| CN114315855B (en) | Curcumenol derivatives, preparation method and application thereof in preparation of anti-inflammatory drugs | |
| CN112940152B (en) | 5-fluorouracil-1-methoxycarbonyl-type cyclodextrin derivative and preparation method thereof | |
| CN111592498B (en) | [2- (5' -fluorouracil) acetic acid-diethyl dithiocarbamic acid ] anhydride and application thereof in preparation of anti-cancer drugs | |
| CN115057831A (en) | Thiadiazole compound and application thereof in preparation of KKLC1 protein inhibitor | |
| CN111153946B (en) | N-azido acetyl-D-mannosamine derivative, preparation method thereof and application thereof in detection of nitroreductase | |
| CN112472683A (en) | Nano diagnosis and treatment agent and preparation method and application thereof | |
| CN114933619B (en) | Thioglycoside column-like analogues and preparation method and application thereof | |
| CN109824655B (en) | Andrographolide compound and preparation method and application thereof | |
| CN102643293A (en) | Cefdinir ternary complex and method for preparing cefdinir by using same | |
| CN107698631B (en) | Lithium hydroxide-based vorinostat derivative and preparation method and application thereof | |
| CN114656477A (en) | SN-38 prodrug responding to hydrogen sulfide as well as preparation method and application thereof | |
| CN101012196A (en) | Fragrant geldanamycin derivative and its preparing method and use | |
| CN115124464A (en) | Quinolinedione sulfonyl piperazine heterozygote and preparation method and application thereof | |
| KR101713678B1 (en) | Novel compounds and pharmaceutical composition for using anticancer drug containing thereof | |
| CN114989214A (en) | Shikonin phosphoramidate heterozygote and synthesis method and application thereof | |
| CN116410203A (en) | Compound with TRK and drug-resistant mutation inhibition activity and application thereof |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |