阅读说明：本技术 一种羽扇豆醇吡啶季铵盐衍生物及其制备方法与应用 (Lupeol pyridine quaternary ammonium salt derivative and preparation method and application thereof ) 是由 韩泽平 何金花 黎毓光 于 2020-07-09 设计创作，主要内容包括：本发明公开一种羽扇豆醇吡啶季铵盐衍生物及其制备方法与应用,属于药物合成领域。本发明提供一种羽扇豆醇吡啶季铵盐衍生物及其在治疗和/或预防癌症方面的应用,其分子式为C<Sub>38</Sub>H<Sub>59</Sub>IN<Sub>2</Sub>O<Sub>2</Sub>,分子量702.1。本发明以羽扇豆醇为起始原料,合成了一种水溶性羽扇豆醇吡啶季铵盐衍生物,药理实验证明该衍生物对多种肿瘤细胞株具有显著的抑制作用,抗癌效果较羽扇豆醇明显,且对正常细胞株毒性作用低,且亲水性好,易溶于水、乙醇、DMSO等溶剂,溶解性优于羽扇豆醇,故具有重大的研发潜力；有望开发成抗肿瘤药物,具有一定的医学价值和市场前景。(The invention discloses lupeol pyridine quaternary ammonium salt derivatives, and a preparation method and application thereof, and belongs to the field of medicine synthesis. The invention provides lupeol pyridine quaternary ammonium salt derivatives and application thereof in treating and/or preventing cancers, wherein the molecular formula of the derivatives is C 38 H 59 IN 2 O 2 Molecular weight 702.1. The invention takes lupeol as an initial raw material to synthesize a water-soluble lupeol pyridine quaternary ammonium salt derivative, and pharmacological experiments prove that the derivative has obvious inhibition effect on various tumor cell strains, has obvious anticancer effect compared with lupeol, has low toxicity on normal cell strains, has good hydrophilicity, is easy to dissolve in solvents such as water, ethanol and DMSO, has better solubility than lupeol, and has great research and development potential; is expected to be developed into antitumor drugs with oneDefinite medical value and market prospect.)

1. A lupeol pyridine quaternary ammonium salt derivative is characterized in that: the structural formula is shown as formula I:

2. use of lupeol pyridiniumnium salt derivative of claim 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment and/or prevention of cancer.

3. Use according to claim 2, characterized in that:

the cancer is selected from bladder cancer, renal cancer, neuroblastoma, liver cancer, nasopharyngeal cancer, prostate cancer, and colon cancer.

4. Use according to claim 2, characterized in that:

the effective concentration of the lupeol pyridine quaternary ammonium salt derivative or the pharmaceutically acceptable salt thereof is 1-80 mu mmol/L of cell suspension, wherein the concentration of the cell suspension is 2.5 × 10⁷cell/L suspension.

5. Use according to claim 4, characterized in that:

the effective concentration of the lupeol pyridine quaternary ammonium salt derivative or the pharmaceutically acceptable salt thereof is 2-80 mu mmol/L of cell suspension.

6. Use according to claim 5, characterized in that:

the effective concentration of the lupeol pyridine quaternary ammonium salt derivative or the pharmaceutically acceptable salt thereof is 2-64 mu mmol/L of cell suspension.

7. Use according to claim 2, characterized in that:

the dosage form of the medicine is oral dosage form or injection dosage form;

the medicament contains one or more pharmaceutically acceptable carriers or excipients.

8. A composition for use in the treatment and/or prevention of cancer, characterized in that: the lupeol pyridine quaternary ammonium salt derivative or the pharmaceutically acceptable salt thereof according to claim 1 as an active ingredient.

9. The process for preparing lupeol pyridinium quaternary derivatives as claimed in claim 1, wherein: the method comprises the following steps:

(1) dissolving the compound 1 in an organic solvent, adding the compound 2, reacting at room temperature, and monitoring by TLC until the compound 1 completely reacts; then adding an organic solvent to dilute the reaction solution, washing with saturated saline solution, drying the organic phase, filtering, and concentrating the filtrate to dryness to obtain a crude product of the compound 3, wherein the crude product is directly used in the next step without further purification;

(2) dissolving the crude product of the compound 3 by using an organic solvent, adding the compound 4 into the solution, and stirring the solution for reaction; and after the reaction is finished, cooling to room temperature, adding an organic solvent to dilute the reaction solution, washing with saturated saline solution, drying the organic phase, filtering, concentrating the filtrate to dryness to obtain a crude product, and purifying the crude product by using a silica gel column chromatography, wherein the eluent is a solvent with a volume ratio of 40: 1 to obtain a white solid compound 5;

(3) taking the compound 5 and dry acetonitrile, stirring and dissolving, adding methyl iodide, and reacting at room temperature after the addition is finished; at the moment, more solids are separated out, the filtration is carried out, and a filter cake is recrystallized by acetonitrile; preparing a tan solid compound A, namely a lupeol pyridine quaternary ammonium salt derivative;

wherein the structural formulas of the compound 1, the compound 3 and the compound 5 are respectively as follows:

the structural formulas of the compound 2 and the compound 4 are respectively as follows:

10. the method of claim 9, wherein:

the organic solvent is dichloromethane;

the concentration of the compound 1 in the step (1) is 100 mmol/L;

the molar ratio of the compound 1 to the compound 2 in the step (1) is 1: 1.1;

the volume ratio of the amount of the organic solvent used for diluting the reaction solution in the step (1) to the amount of the organic solvent used for dissolving the compound 1 is 1: 1;

the dosage of the compound 4 in the step (2) is determined according to the molar ratio of the compound 1 to the compound 4, namely 1: 1.2, adding;

the volume ratio of the amount of the organic solvent used for diluting the reaction solution in the step (2) to the amount of the organic solvent used for dissolving the compound 3 is 1: 1;

the molar ratio of the compound 5 to the methyl iodide in the step (3) is 1: 7.3;

the room-temperature reaction time in the step (1) is 2 hours;

the stirring reaction condition in the step (2) is that stirring reaction is carried out for 17 hours at 40 ℃;

the reaction time at room temperature in the step (3) is 17 h.

Technical Field

The invention belongs to the field of drug synthesis, and relates to lupeol pyridine quaternary ammonium salt derivatives, and a preparation method and application thereof.

Background

Cancer, i.e., malignant tumor, is a serious disease that endangers human health caused by multigenic mutations, multifactorial causes, and multistage development. Recent data show 18078957 cases of new cancer cases and 9555027 cases of cancer death in 2018 worldwide. Therefore, various works for preventing and treating cancer are actively carried out, and how to effectively treat cancer has become a urgent necessity. The malignant tumor treatment method mainly comprises surgical treatment, radiotherapy, chemotherapy, biological treatment and the like, the chemotherapy is still one of the main means for treating the tumor at present and in a long term in the future, but the side effect caused by chemotherapy seriously reduces the life quality of patients. In the face of a plurality of tumor diseases at present, the existing antitumor drugs far from meeting the requirements are urgently needed to be researched and developed.

Lupeol (lupeol) is a pentacyclic triterpene compound derived from herbal plants such as Lupeol, fructus Mangifera Indicae and herba Taraxaci, and has a molecular formula of C₃₀H₅₀O, molecular weight 426.7174, has effects of anti-inflammatory, antioxidant, anti-infection, anti-hyperglycemia, anti-asthma, antirheumatic, protecting heart, protecting nerve, protecting liver and regulating immunity, etc., and suggests that lupeol has wide pharmacological activity and multiple action mechanisms and targets. In the anti-tumor research, lupeol has good inhibitory effect on various tumor cells, including colorectal cancer, prostate cancer, liver cancer, lung cancer, bladder cancer and the like, and the mechanism of lupeol relates to the change of various intracellular signal pathways. However, lupeol is insoluble in water and, although it is miscible with diethyl ether, benzene, petroleum ether, and hot ethanol, its solubility is not good. In research experiments, warm absolute ethanol and Dimethylsulfoxide (DMSO) were often used in a 1: 1 as a solvent, but the dissolution effect is still not ideal, drug crystals are easy to precipitate, the research on the anti-tumor aspect of the drug is limited, and the development of the drug to clinical application is further hindered.

Disclosure of Invention

In order to overcome the disadvantages and shortcomings of the prior art, the invention provides a lupeol pyridine quaternary ammonium salt derivative. The derivative is prepared by connecting lupeol and hydrophilic group, and has molecular formula of C₃₈H₅₉IN₂O₂Molecular weight 702.1; the compound has good hydrophilicity, and the compound is tested for anti-tumor activity, thereby providing the lupeol derivative with certain medicinal value.

The invention also aims to provide application of the lupeol pyridine quaternary ammonium salt derivative.

The invention carries out the activity detection experiment of human tumor cells and normal epithelial cells on the synthesized lupeol pyridine quaternary ammonium salt derivative, and the result shows that the lupeol pyridine quaternary ammonium salt derivative can obviously inhibit the proliferation of various tumor cells, has higher inhibition rate than lupeol, has small toxicity to the normal epithelial cells of human, is easy to dissolve in solvents such as water, ethanol, DMSO and the like, has better solubility than lupeol, and has great research and development potential.

The invention also aims to provide a preparation method of the lupeol pyridine quaternary ammonium salt derivative.

The purpose of the invention is realized by the following technical scheme:

the invention provides a lupeol pyridine quaternary ammonium salt derivative, which has a structural formula shown as a formula I:

the invention also provides application of the lupeol pyridine quaternary ammonium salt derivative or the pharmaceutically acceptable salt thereof in preparing a medicament for treating and/or preventing cancer.

Preferably, the cancer is selected from bladder cancer, kidney cancer, neuroblastoma, liver cancer, nasopharyngeal cancer, prostate cancer, colon cancer, etc.;

the effective concentration of the lupeol pyridine quaternary ammonium salt derivative or the pharmaceutically acceptable salt thereof is 1-80 mu mmol/L of cell suspension, further 2-80 mu mmol/L of cell suspension, and further 2-64 mu mmol/L of cell suspension, wherein the concentration of the cell suspension is 2.5 × 10⁷cell/L suspension.

The dosage form of the medicine is oral dosage form or injection dosage form;

the medicament contains one or more pharmaceutically acceptable carriers or excipients.

A composition for treating and/or preventing cancer, wherein the active ingredient is the lupeol pyridine quaternary ammonium salt derivative or the pharmaceutically acceptable salt thereof.

The invention also provides a preparation method of the lupeol pyridine quaternary ammonium salt derivative, which comprises the following steps:

(1) dissolving the compound 1 in an organic solvent, adding the compound 2, reacting at room temperature, and monitoring by TLC until the compound 1 completely reacts; then adding an organic solvent to dilute the reaction solution, washing with saturated saline solution, drying the organic phase, filtering, and concentrating the filtrate to dryness to obtain a crude product of the compound 3, wherein the crude product is directly used in the next step without further purification;

(2) dissolving the crude product of the compound 3 by using an organic solvent, adding the compound 4 into the solution, and stirring the solution for reaction; cooling to room temperature after the reaction is finished, adding an organic solvent to dilute the reaction solution, washing with saturated saline solution, drying the organic phase, filtering, concentrating the filtrate to dryness to obtain a crude product, purifying the crude product by using a silica gel column chromatography, and obtaining a white-like solid compound 5 by using a dichloromethane-methanol (40: 1) eluent, wherein the total yield of the two steps is 36.86%;

(3) taking the compound 5 and dry acetonitrile, stirring and dissolving, adding methyl iodide, and reacting at room temperature after the addition is finished; at the moment, more solids are separated out, the filtration is carried out, and a filter cake is recrystallized by acetonitrile; preparing a tan solid compound A, namely a lupeol pyridine quaternary ammonium salt derivative; the yield of this step was 36.65%.

Wherein the structural formulas of the compound 1, the compound 3 and the compound 5 are respectively as follows:

the structural formulas of the compound 2 and the compound 4 are respectively as follows:

preferably, the organic solvent is dichloromethane;

preferably, the concentration of the compound 1 in the step (1) is 100 mmol/L;

preferably, the molar ratio of compound 1 to compound 2 in step (1) is 1: 1.1;

preferably, the reaction time at room temperature in the step (1) is 2 h;

preferably, the volume ratio of the amount of the organic solvent used for diluting the reaction solution in the step (1) to the amount of the organic solvent used for dissolving the compound 1 is 1: 1.

preferably, the compound 4 in the step (2) is used in an amount of 1: 1.2, adding;

preferably, the stirring reaction condition in the step (2) is that the stirring reaction is carried out for 17 hours at 40 ℃;

preferably, the volume ratio of the amount of the organic solvent used for diluting the reaction solution in the step (2) to the amount of the organic solvent used for dissolving the compound 3 is 1: 1.

preferably, the molar ratio of the compound 5 to the methyl iodide in the step (3) is 1: 7.3.

preferably, the reaction time at room temperature in the step (3) is 17 h.

Compared with the prior art, the invention has the following advantages and effects:

the invention provides lupeol pyridine quaternary ammonium salt derivatives shown as a formula I and application thereof in treating and/or preventing cancers. The invention takes lupeol as a starting material to synthesize a water-soluble lupeol pyridine quaternary ammonium salt derivative, and pharmacological experiments prove that the derivative has obvious inhibition effect on various tumor cell strains, has obvious anticancer effect compared with lupeol, has low toxicity on normal cell strains, is expected to be developed into an antitumor drug, and has certain medical value and market prospect.

Detailed Description

The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.

The experimental procedures for specific experimental conditions not specified in the following examples are generally performed according to conventional experimental conditions or according to experimental conditions recommended by the manufacturers. The materials, reagents and the like used are, unless otherwise specified, reagents and materials obtained from commercial sources.