阅读说明：本技术 一类甲硝唑接肌氨酰-组氨酸二肽化合物及其制备与应用 (Metronidazole-sarcosyl-histidine dipeptide compound and preparation and application thereof ) 是由 马烨 张旭萍 于 2019-09-29 设计创作，主要内容包括：本发明公开了一类甲硝唑接肌氨酰-组氨酸二肽化合物：2-(2-甲基-5-硝基-1H-咪唑基)乙基肌氨酰组氨酸,分子式为C<Sub>15</Sub>H<Sub>21</Sub>N<Sub>7</Sub>O<Sub>5</Sub>,其综合利用组氨酸和肌氨酸的优势,将组氨酸和肌氨酸合成为Sar-His二肽,并修饰到甲硝唑上,降低了幽门杆菌的耐药性,增强甲硝唑类药物对幽门螺旋杆菌的抑制作用,优化甲硝唑对幽门螺旋杆菌的抑制效果,降低甲硝唑对人体的副反应,提升人体免疫力,且原料易获取,合成工艺简单,易操作,大大增加了甲硝唑修饰物的市场前景,具有很强的实用性和广泛适用性。(The invention discloses a metronidazole-sarcosyl-histidine dipeptide compound: 2- (2-methyl-5-nitro-1H-imidazolyl) ethylsarcosyl histidine with the molecular formula C 15 H 21 N 7 O 5 The advantages of histidine and sarcosine are comprehensively utilized, the histidine and the sarcosine are synthesized into Sar-His dipeptide and are modified on metronidazole, the drug resistance of helicobacter pylori is reduced, the inhibiting effect of metronidazole drugs on the helicobacter pylori is enhanced, the inhibiting effect of the metronidazole on the helicobacter pylori is optimized, the side reaction of the metronidazole on a human body is reduced, the immunity of the human body is improved, the raw materials are easy to obtain, the synthesis process is simple, the operation is easy, and the advantages of the histidine and the sarcosine are greatly increasedThe nitre modifier has market prospect, strong practicability and wide applicability.)

1. A metronidazole-sarcosyl-histidine dipeptide compound is characterized in that the molecular formula is as follows: c₁₅H₂₁N₇O₅The structural formula is as follows:

2. the preparation of a metronidazole sarcosyl-histidine dipeptide compound according to claim 1, characterised in that the reaction is as follows:

3. the preparation of a metronidazole sarcosyl-histidine dipeptide compound according to claim 1, comprising the steps of:

s1, dissolving metronidazole (compound 1) and sarcosyl-histidine dipeptide (compound 2) in a proper amount of dichloromethane solution, simultaneously adding a certain amount of DCC and DMAP, stirring for reaction, and then distilling under reduced pressure to obtain a crude product;

s2, purifying the crude product by column chromatography to obtain white powder (compound 3).

4. The preparation of a class of metronidazole sarcosyl-histidine dipeptide compounds according to claim 3, wherein the molar ratio of metronidazole (compound 1) and Phe-Lys dipeptide (compound 2) in step S1 is 1: 1.1.

5. The preparation of a metronidazole sarcosyl-histidine dipeptide compound according to claim 3 where the DCC is 1.5equiv and the DMAP is 0.3 equiv.

6. The preparation of a class of metronidazole sarcosyl-histidine dipeptide compounds according to claim 3, wherein in step S1 the reaction is followed by TLC.

7. The preparation of a metronidazole-sarcosyl-histidine dipeptide compound according to claim 2 or 3, wherein the metronidazole-sarcosyl-histidine dipeptide is synthesized by a polypeptide synthesizer;

the Fmoc/tBu solid-phase polypeptide synthesis method is utilized, then 20 wt% of piperidine and dimethylformamide are used for removing Fmoc, and the prepared crude product is purified by column chromatography to obtain a pure product Sar-His.

8. The use of a class of metronidazole sarcosyl-histidine dipeptide compounds according to claim 1 in the inhibition of helicobacter pylori.

Technical Field

The invention relates to a metronidazole-sarcosyl-histidine dipeptide compound, in particular to a metronidazole-sarcosyl-histidine dipeptide compound and preparation and application thereof.

Background

Metronidazole (metronidazo1e), also known as metronidazole, is a nitroimidazole antibiotic, has powerful bactericidal action on gram-positive and gram-negative anaerobic bacteria and bacteroides fragilis, and is a clinically common anti-infection basic drug. With the clinical wide application of metronidazole and the continuous and deep research on the pharmacological mechanism of metronidazole, various documents report that the adverse reaction of metronidazole is rare.

Helicobacter pylori (Hp) is a gram-negative, sigmoidal or arcuately curved bacterium, and it has been shown through epidemiological, clinical and pathological studies that Hp can induce chronic gastritis, peptic ulcer, gastric mucosa-associated lymphoid tissue (muco-associated lymphoma) lymphoma, intestinal gastric cancer in humans. The world health organization has listed helicobacter pylori as a class I carcinogen, which is a major causative factor of gastric cancer.

Therefore, there is a need for metronidazole modifications that improve its pharmacological action against gram-positive and gram-negative anaerobes, especially against helicobacter pylori, and reduce its adverse effects.

Disclosure of Invention

In order to solve the defects of the prior art, the invention aims to provide a metronidazole sarcosyl-histidine dipeptide compound capable of effectively enhancing the inhibition effect of metronidazole drugs on helicobacter pylori and a preparation method thereof.

In order to achieve the above object, the present invention adopts the following technical solutions:

a metronidazole-sarcosyl-histidine dipeptide compound has a molecular formula as follows: c₁₅H₂₁N₇O₅The structural formula is as follows:

the preparation of the metronidazole-sarcosyl-histidine dipeptide compound has the following reaction formula:

the preparation method of the metronidazole-sarcosyl-histidine dipeptide compound comprises the following steps:

s1, dissolving metronidazole (compound 1) and sarcosyl-histidine dipeptide (compound 2) in a proper amount of dichloromethane solution, simultaneously adding a certain amount of DCC and DMAP, stirring for reaction, and then distilling under reduced pressure to obtain a crude product;

s2, purifying the crude product by column chromatography to obtain white powder (compound 3).

Further, in the above step S1, the molar ratio of metronidazole (compound 1) to Phe-Lys dipeptide (compound 2) is 1: 1.1.

Further, the DCC was 1.5equiv, and DMAP was 0.3 equiv.

Further, in the above step S1, the reaction was followed by TLC.

The oxazole-linked sarcosyl-histidine dipeptide compound is synthesized by a polypeptide synthesizer;

the Fmoc/tBu solid-phase polypeptide synthesis method is utilized, then 20 wt% of piperidine and dimethylformamide are used for removing Fmoc, and the prepared crude product is purified by column chromatography to obtain a pure product Sar-His.

The metronidazole-sarcosyl-histidine dipeptide compound is applied to inhibiting helicobacter pylori.

The invention has the advantages that:

the metronidazole-sarcosyl-histidine dipeptide compound comprehensively utilizes the advantages of histidine and sarcosine, synthesizes the histidine and the sarcosine into Sar-His dipeptide, modifies the Sar-His dipeptide on the metronidazole, enhances the antibacterial effect of the metronidazole, particularly enhances the inhibition effect of the Sar dipeptide on helicobacter pylori, reduces the side effect of the metronidazole on a human body, and improves the immunity of the human body.

Histidine is an imidazolyl structure, can form a coordination compound with ferrous ions, promotes the absorption of iron, and can be used for preventing and treating anemia. More importantly, the histidine can reduce the acidity of gastric juice, relieve the pain of gastrointestinal surgery, reduce the burning sensation of the stomach and inhibit the digestive tract ulcer; it has great curative effect on relieving gastric ulcer and enteritis caused by helicobacter pylori. Creatine exists in human body in the form of phosphocreatine, and can continuously promote synthesis of ATP required by human body.

The metronidazole sarcosyl-histidine dipeptide compound has the advantages of easily obtained raw materials, simple synthesis process and easy operation, effectively reduces the drug resistance of helicobacter pylori, enhances the inhibition effect of metronidazole drugs on the helicobacter pylori, optimizes the inhibition effect of metronidazole on the helicobacter pylori, greatly increases the market prospect of metronidazole modifiers, and has strong practicability and wide applicability.

Detailed Description

The present invention will be described in detail with reference to the following embodiments.

The reagents used in the examples of the present invention are all commercially available.

DCC is dicyclohexylcarbodiimide and DMAP is 4-dimethylaminopyridine.

A metronidazole-sarcosyl-histidine dipeptide compound, namely 2- (2-methyl-5-nitro-1H-imidazolyl) ethylsarcosyl histidine (compound 3), has a molecular formula as follows: c₁₅H₂₁N₇O₅The structural formula is as follows:

the reaction formula is as follows:

the preparation method comprises the following steps:

s1, dissolving metronidazole (compound 1)200mg and sarcosyl-histidine dipeptide (compound 2)291mg in a proper amount of dichloromethane solution according to a molar ratio of 1:1.1, simultaneously adding 1.5equiv DCC and 0.3equiv DMAP, stirring at room temperature for reaction, tracking the reaction by TLC, and after the reaction is completed, distilling a crude product under reduced pressure;

s2, purification of the crude product by column chromatography gave a white powder (compound 3) (798mg, 88% yield).¹H NMR(500MHz,CDCl₃)δ7.94(d,J＝12.8Hz,1H),7.91(s,1H),7.54(dd,J＝7.6,1.7Hz,1H),6.99(dd,J＝6.6,1.8Hz,1H),6.90(dd,J＝7.5,6.6Hz,1H),4.65–4.53(m,3H),4.47(ddd,J＝7.7,7.1,1.8Hz,2H),4.38(tq,J＝6.9,3.3Hz,1H),3.31–3.09(m,4H),2.50(d,J＝3.3Hz,3H),2.43(s,3H)。

Oxazole-linked sarcosyl-histidine dipeptide (Sar-His) compounds, which can be synthesized by a polypeptide synthesizer; the Fmoc/tBu solid-phase polypeptide synthesis method is utilized, then 20 wt% of piperidine and dimethylformamide are used for removing Fmoc, and the prepared crude product is purified by column chromatography to obtain a pure product Sar-His.

Detecting the effect of the metronidazole-sarcosyl-histidine dipeptide compound on inhibiting helicobacter pylori:

(I) test materials

Culture medium: nutrient agar;

fresh defibrinated horse blood;

starch;

mixing antibiotics: vancomycin, a xanthamine synergist TMP, amphotericin, polymyxin;

experimental strains: helicobacter pylori;

the medicine solvent is as follows: and (3) ethanol.

(II) culturing helicobacter pylori

The culture conditions are as follows: the temperature is 37 ℃, the pH value is 7.0-7.2, and the oxygen content is 2-8%;

culture medium: adding a proper amount of horse blood, a proper amount of nutrient agar culture medium mixed with antibiotics and 1% of starch;

culturing time: 3-5 days.

(III) the result of the detection

The experimental components were divided into four groups: filter paper without any drug, filter paper with metronidazole (10mg/mL), filter paper with compound 2(10mg/mL dipeptide complex), filter paper with compound 3(10mg/mL metronidazole-sarcosyl-histidine dipeptide complex).

After the bacteria are cultured, the size of the inhibition zone is measured by a conventional filter paper diffusion method to carry out a drug sensitivity experiment, and the experiment results are shown in the following table 1:

TABLE 1 inhibitory Effect of different Compounds on helicobacter pylori

Components	Has no drug effect	Compound 2	Metronidazole	Compound 3
					Size/ratio of zone of inhibition	0	0.4	1	1.5

As can be seen from the experimental results of table 1 above, compound 3, i.e., the metronidazole sarcosyl-histidine dipeptide compound of the present invention, has an obvious inhibitory effect on helicobacter pylori than metronidazole.

The foregoing illustrates and describes the principles, general features, and advantages of the present invention. It should be understood by those skilled in the art that the above embodiments do not limit the present invention in any way, and all technical solutions obtained by using equivalent alternatives or equivalent variations fall within the scope of the present invention.