阅读说明：本技术 糖环修饰的核苷类似物、制备方法及其在制备具有抗疲劳功能产品中的应用 (Sugar ring modified nucleoside analogue, preparation method and application thereof in preparation of product with anti-fatigue function ) 是由 徐亮 朱慧敏 曾万波 史卫国 于 2021-10-27 设计创作，主要内容包括：本发明涉及制药技术领域,具体涉及糖环修饰的核苷类似物、制备方法及其在制备具有抗疲劳功能产品中的应用。本发明首次发现了,多种现有的AICAR类似物和腺苷类似物也能够用于提升体能,发挥抗疲劳功能。本发明还进一步合成了多种新的AICAR类似物和腺苷类似物,并提供了含有上述AICAR类似物和/或腺苷类似物的药物,给出了所述药物在应用时的有效剂量为0.0094～4.7mmol/kg/d。(The invention relates to the technical field of pharmacy, in particular to a nucleoside analogue modified by sugar ring, a preparation method and application thereof in preparing products with anti-fatigue function. The invention discovers for the first time that a plurality of existing AICAR analogues and adenosine analogues can also be used for improving physical ability and playing a role in resisting fatigue. The invention further synthesizes a plurality of novel AICAR analogs and adenosine analogs, provides a medicament containing the AICAR analogs and/or adenosine analogs, and provides that the effective dose of the medicament in application is 0.0094-4.7 mmol/kg/d.)

1. The application of nucleoside analogues modified by sugar rings in preparing products with anti-fatigue functions.

2. Use according to claim 1, wherein the product comprises a medicament with anti-fatigue properties.

3. The use of claim 1, wherein the sugar ring modified nucleoside analog comprises a lipophilic nucleoside analog having the molecular structure:

wherein R is₁、R₂、R₃Is selected from-OH, -OCO (CH)₂)_nCH₃One of halogen, alkyl or aromatic group; r₄Including heterocyclic compounds; wherein n is more than or equal to 1.

4. Use according to claim 3, wherein said heterocyclic compound is selected fromQuinoline, pyrrole, furan, thiophene or indole.

5. Use according to claim 3, wherein R is₂And R₃Are connected to form a ring structure;

preferably, at least one of a halogen, an alkyl or an aromatic group is attached to the cyclic structure.

6. A sugar ring-modified nucleoside analog, comprising compound a having the molecular structural formula:

wherein R is₁、R₂And R₃All are anhydride substituents with carbon number more than or equal to 3, R₄Is selected fromOne of quinoline, pyrrole, furan, thiophene or indole;

or, compound B, having the molecular structure:

wherein R is₂And R₃Cyclic, the cyclic structure of which is linked to an aromatic group, R₁is-OH, R₄Is selected fromOne of quinoline, pyrrole, furan, thiophene or indole;

or, compound C, having the molecular structure:

wherein R is₂And R₃Form a ring, a halogenated aromatic group is connected to the ring structure, R₁is-OH, R₄Is composed ofOne of quinoline, pyrrole, furan, thiophene or indole;

preferably, the sugar ring modified nucleoside analogue comprises compound D, the molecular structural formula of which is:

wherein R is₁、R₂And R₃Is an anhydride substituent with 4 or more carbon atoms; r₄Is composed of

Preferably, the sugar ring modified nucleoside analog comprises compound E, the molecular structural formula of which is:

wherein R is₂And R₃Form a ring, an aromatic group is connected on the ring structure, R₁is-OH, R₄Is composed of

Preferably, the sugar ring modified nucleoside analogue comprises compound F, the molecular structural formula of which is:

wherein R is₂And R₃Form a ring, a halogenated aromatic group is connected to the ring structure, R₁is-OH, R₄Is composed of

7. The method for preparing a sugar ring-modified nucleoside analog according to claim 6, wherein the sugar ring-modified nucleoside analog is compound D, and the method for preparing the compound D comprises the steps of carrying out nucleophilic substitution reaction on AICAR or adenosine and acid anhydride under the condition of anhydrous pyridine, wherein every 1mol of adenosine is reacted with 3-4 mol of acid anhydride, and the acid anhydride comprises acetic anhydride, propionic anhydride, butyric anhydride or isobutyric anhydride;

or the sugar ring modified nucleoside analogue is a compound E, and the preparation method of the compound E comprises the step of preparing R in the AICAR analogue₂And R₃After cyclization, the cyclic structure and aromatic aldehyde are subjected to addition reaction, so that the cyclic structure is connected with aromatic groups, 1mol of AICAR analogue reacts with 1-2 mol of aromatic aldehyde, the aromatic aldehyde comprises p-methoxybenzaldehyde, 3' -methylbenzaldehyde, m-fluorobenzaldehyde, m-chlorobenzaldehyde or m-bromobenzaldehyde, and the addition reaction is preferably carried out in the presence of anhydrous zinc chlorideOccurs in the tetrahydrofuran reaction system;

or the sugar ring modified nucleoside analogue is compound F, and the preparation method of the compound F comprises the step of preparing R in the AICAR analogue₂And R₃After cyclization, performing addition reaction on the cyclic structure and hydrogen halide to ensure that the cyclic structure is connected with a halogen group, and reacting 1-2 mol of hydrogen halide including hydrogen fluoride or hydrogen bromide with each 1mol of AICAR analogue; the addition reaction preferably takes place in a tetrahydrofuran reaction system containing anhydrous zinc chloride.

8. A pharmaceutical comprising the sugar ring-modified nucleoside analog according to claim 6, or the sugar ring-modified nucleoside analog obtained by the production method according to claim 7, and a pharmaceutically acceptable carrier or excipient.

9. The medicament of claim 8, wherein the medicament is in a dosage form selected from the group consisting of an injection, a tablet, a pill, a capsule, a suspension, a granule, a spray, and an emulsion.

10. The effective dose of the medicine of claim 8 or 9 is 0.00784-1.96 mmol/kg/d;

preferably, it is 0.0196 mmol/kg/d.

Technical Field

The invention relates to the technical field of food and medicine, in particular to a sugar ring modified nucleoside analogue, a preparation method and application thereof in preparing a product with an anti-fatigue function.

Background

Skeletal muscle is an adaptive tissue composed of a plurality of muscle fibers that vary in their metabolic and contractile properties, including the oxidative slow muscle (type I), the mixed oxidative/glycolytic fast muscle (type IIa) and the glycolytic fast muscle (type IIb). Type I fibers preferentially express enzymes that oxidize lipids, contain slow substitutes for contractile proteins, and are more fatigue resistant than glycolytic fibers. Type IIa fibres preferentially metabolize glucose and express the fast isoform of contractile protein. Endurance exercise training triggers skeletal muscle remodeling programs that gradually enhance performance of athletes, such as marathons, mountain climbers, and bicyclists. These performance-improving indications may also prevent obesity and related metabolic disorders of skeletal muscle, and are resistant to muscle wasting.

The increasing sedentary lifestyle and unprecedented excessive caloric food intake in modern society has strongly driven the prevalence of many diseases, including obesity, cardiovascular disease, metabolic syndrome, and other chronic diseases. The development of drugs associated with these lifestyle-related diseases is extremely attractive: these diseases affect a large number of patients and require long-term administration of drugs for prevention and treatment over a period of years or even decades. The underlying mechanisms of therapeutic effect in physical activity remain unclear as exercise induces many different plasticity changes in skeletal muscle and other sites. While physical exercise remains the best solution, the development of muscle-targeted "exercise mimics" will soon provide a medicinal alternative to overcome the increasingly sedentary lifestyle. Narkar et al show that some phenotypic adaptation of muscles important for human health can be mimicked by drug treatment of mice.

Exercise training activates many transcriptional regulators in skeletal muscle as well as serine/threonine kinases, thereby facilitating reprogramming of the metabolism. Serine/threonine kinases are the most widely known ones, AMP-activated protein kinase (AMPK), a major regulator of cellular and body metabolism, and their function is conserved in all eukaryotes. In mammals, AMPK has been shown to contribute to glucose homeostasis, appetite and motor physiology. Peroxisome proliferator-activated receptor gamma coactivator 1(PGC-1), a cotranscription regulator, has been identified as a major regulator of Brown Adipose Tissue (BAT) and mitochondrial biogenesis. In recent years, activation of AMP-activated protein kinase (AMPK) has been shown to modulate the expression and activity of PGC-1 α. The AMPK is activated, the transcription activity of PGC-1 alpha mRNA can be increased, the transcription of genes related to the fiber of the slow muscle of skeletal muscle is promoted, and the improvement of the exercise endurance is promoted. Narkar et al showed that activation of the downstream substrate PGC-1 with the AMPK agonist adenosine analog AICAR resulted in significant muscle reprogramming, leading to a more robust oxidative phenotype with a 44% increase in tolerance 4 weeks after dosing in untrained naive mice. This result suggests that the AMPK-PGC-1. alpha. pathway shows important significance in motor simulation.

AICAR, collectively known as 5-amino imidazole-4-carboxamide1- β -D-ribofuranoside, also known as AICariboside, is an activator of AMP-activated protein kinase (AMPK) which can permeate cell membranes. AMPK is a key protein for metabolic regulation, and when the ratio of AMP to ATP is up-regulated when the energy supply is insufficient, AMPK is activated and anabolism is inhibited. AICAR can activate AMPK, but does not affect the levels of ATP, ADP, and AMP. At the cellular or animal level, AICAR can promote non-insulin dependent glucose uptake in skeletal muscle by activating AMPK. AICAR-induced skeletal muscle glucose uptake cannot be blocked by inhibitors of PI 3K.

Adenosine is a cellular biological term, and refers to a compound in which N-9 of adenine and C-1 of D-ribose are linked by a β -glycosidic bond, and its phosphate is adenylic acid. Adenosine is an endogenous nucleoside distributed throughout human cells, can directly enter cardiac muscle to generate adenosine through phosphorylation, participates in cardiac muscle energy metabolism, and also participates in dilating coronary vessels to increase blood flow.

With the commercial popularization of AICAR and adenosine, the prospect of the anti-fatigue function of various analogs of AICAR and adenosine is gradually shown, but at present, no clear report exists on which analogs can be used in anti-fatigue products as substitute products of AICAR and adenosine.

In view of this, the invention is particularly proposed.

Disclosure of Invention

The invention aims to provide application of a sugar ring modified nucleoside analogue in preparing a product with an anti-fatigue function, and the sugar ring modified nucleoside analogue with higher anti-fatigue capability replaces the currently adopted AICAR and adenosine products so as to meet the requirement of anti-fatigue products in the field of daily commodities.

The second object of the present invention is to provide various sugar ring lipophilically modified nucleoside analogs, which have higher fatigue resistance and can be commercially circulated, compared to the existing AICAR and adenosine.

The third purpose of the invention is to provide a preparation method of the nucleoside analogues modified by multiple sugar rings, wherein the preparation method is simple and feasible and is suitable for industrial popularization.

It is a final object of the present invention to provide a pharmaceutical or functional food which can achieve a significant anti-fatigue effect by using a lower effective dose of a sugar ring-modified nucleoside analog, thereby greatly expanding the existing application fields of AICAR and adenosine to meet the needs of more groups.

In order to solve the technical problems and achieve the purpose, the invention provides the following technical scheme:

in a first aspect, the invention provides an application of nucleoside analogues modified by sugar rings in preparation of products with anti-fatigue functions.

In an alternative embodiment, the product comprises a medicament with anti-fatigue properties.

In alternative embodiments, the sugar ring modified nucleoside analog comprises a lipophilic nucleoside analog having the molecular structure:

wherein R is₁、R₂、R₃including-OH, -OCO (CH)₂)_nCH₃One of halogen, alkyl or aromatic group; r₄Including heterocyclic compounds; wherein n is more than or equal to 1.

In an alternative embodiment, the heterocyclic compound is selected from Quinoline, pyrrole, furan, thiophene or indole.

In an alternative embodiment, said R₂And R₃The connection between them forms a ring structure.

Preferably, at least one of a halogen, an alkyl or an aromatic group is attached to the cyclic structure.

In a second aspect, the present invention provides a sugar ring modified nucleoside analog, which includes compound a, having a molecular structural formula:

wherein R is₁、R₂And R₃All are anhydride substituents with carbon number more than or equal to 3, R₄Is selected fromOne of quinoline, pyrrole, furan, thiophene or indole;

or, compound B, having the molecular structure:

wherein R is₂And R₃Cyclic, the cyclic structure of which is linked to an aromatic group, R₁is-OH, R₄Is selected fromOne of quinoline, pyrrole, furan, thiophene or indole;

or, compound C, having the molecular structure:

wherein R is₂And R₃Form a ring, a halogenated aromatic group is connected to the ring structure, R₁is-OH, R₄Is composed ofQuinoline, pyrrole, furan, thiophene or indole.

Preferably, the sugar ring modified nucleoside analogue comprises compound D, the molecular structural formula of which is:

wherein R is₁、R₂、R₃Is an anhydride substituent with 4 or more carbon atoms; r₄Is composed of

Preferably, the sugar ring modified nucleoside analog comprises compound E, the molecular structural formula of which is:

wherein R is₂And R₃Form a ring, an aromatic group is connected on the ring structure, R₁is-OH, R₄Is composed of

Preferably, the sugar ring modified nucleoside analogue comprises compound F, the molecular structural formula of which is:

wherein R is₂And R₃Form a ring, a halogenated aromatic group is connected to the ring structure, R₁is-OH, R₄Is composed of

In a third aspect, the invention provides a preparation method of the sugar ring modified nucleoside analogue, wherein the sugar ring modified nucleoside analogue is a compound D, and the preparation method of the compound D comprises the steps of enabling AICAR or adenosine to undergo nucleophilic substitution reaction with acid anhydride under the condition of anhydrous pyridine, and enabling every 1mol of adenosine to react with 3-4 mol of acid anhydride, wherein the acid anhydride comprises acetic anhydride, propionic anhydride, butyric anhydride or isobutyric anhydride;

or the sugar ring modified nucleoside analogue is a compound E, and the preparation method of the compound E comprises the step of preparing R in the AICAR analogue₂And R₃After cyclization, performing addition reaction on the cyclic structure and aromatic aldehyde to ensure that the cyclic structure is connected with aromatic groups, wherein 1mol of AICAR analogue reacts with 1-2 mol of aromatic aldehyde, the aromatic aldehyde comprises p-methoxybenzaldehyde, 3' -methylbenzaldehyde, m-fluorobenzaldehyde, m-chlorobenzaldehyde or m-bromobenzaldehyde, and the addition reaction is preferably performed in a tetrahydrofuran reaction system containing anhydrous zinc chloride;

or the sugar ring modified nucleoside analogue is compound F, and the preparation method of the compound F comprises the step of preparing AICAR analogueIn R₂And R₃After cyclization, performing addition reaction on the cyclic structure and hydrogen halide to ensure that the cyclic structure is connected with a halogen group, and reacting 1-2 mol of hydrogen halide including hydrogen fluoride or hydrogen bromide with each 1mol of AICAR analogue; the addition reaction preferably takes place in a tetrahydrofuran reaction system containing anhydrous zinc chloride.

In a fourth aspect, the present invention provides a medicament comprising the sugar ring-modified nucleoside analog according to the foregoing embodiment, or comprising the sugar ring-modified nucleoside analog obtained by the preparation method according to the foregoing embodiment, and a pharmaceutically acceptable carrier or excipient, in combination with the second and third aspects.

In alternative embodiments, the pharmaceutical dosage form comprises an injection, a tablet, a pill, a capsule, a suspension, a granule, a spray, or an emulsion.

In a fifth aspect, the invention provides, in combination with the fourth aspect, the effective dose of the medicament according to the previous embodiment or the functional food according to the previous embodiment in application is 0.00784-1.96 mmol/kg/d.

Preferably, it is 0.0196 mmol/kg/d.

The invention discovers for the first time that besides AICAR and adenosine, a plurality of existing AICAR analogues and adenosine analogues can be used for improving physical ability and playing a role in resisting fatigue.

The invention further synthesizes a plurality of novel AICAR analogs and adenosine analogs, wherein the AICAR analogs and the adenosine analogs contain acid anhydride groups, or cyclic structures connected with aromatic groups, or cyclic structures connected with halogenated aromatic groups, and the three functional groups can increase the lipophilicity of adenosine compounds, improve the stability of systemic circulation, change the hydrophilic-lipophilic balance value of the drugs, and better permeate cell membranes, thereby improving the bioavailability of the drugs.

The invention also provides a medicament and a functional food containing the AICAR analogue and/or adenosine analogue, and provides that the effective dose of the medicament and the functional food is 0.00784-1.96 mmol/kg/d when the medicament and the functional food are applied. Is far less than the effective dose of the existing AICAR and adenosine products, has more remarkable anti-fatigue effect, realizes the aim of small amount and high efficiency, is more suitable for commercial promotion, and has higher safety.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a molecular structural formula of a compound provided in embodiments 1 to 6 of the present invention;

FIG. 2 is a molecular structural formula of a compound provided in example 7 of the present invention;

FIG. 3 is a molecular structural formula of the compound provided in examples 8 to 13 of the present invention;

FIG. 4 is a molecular structural formula of a compound provided in examples 14 and 15 of the present invention;

FIG. 5 is a molecular structural formula of a compound provided in examples 16 and 17 of the present invention;

FIG. 6 shows the distance to exhaustion of each group of mice in Experimental example 1.1 of the present invention;

FIG. 7 shows the distance to exhaustion of each group of mice in Experimental example 1.2 of the present invention;

FIG. 8 is a comparison of the distance to exhaustion for each group of mice in Experimental example 1.3 of the present invention;

FIG. 9 shows the time to exhaustion of the swimming test for each group of mice in Experimental example 2.1 of the present invention;

FIG. 10 shows the time to exhaustion of the swimming test for each group of mice in Experimental example 2.2 of the present invention;

FIG. 11 is a graph showing the average maximum heights of the groups of mice in Experimental example 3 of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.

Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.

Some embodiments of the invention are described in detail below with reference to the accompanying drawings. The embodiments described below and the features of the embodiments can be combined with each other without conflict.

Examples 1 to 6

Embodiments 1-6 of this group each provide an AICAR analog having the general structural formula:specific molecular structural formulas of examples 1 to 6 are shown as 1a to 1f in FIG. 1, respectively.

Example 1 provides AICAR analogs wherein R₁is-OH, R₂And R₃Form a ring, R₄Is composed ofAs shown at 1a in fig. 1.

Example 2 provides AICAR analogs wherein R₁、R₂And R₃Are all acetic anhydride, R₄Is composed ofAs shown at 1b in fig. 1.

AICAR like that given in example 3In which R is₁、R₂And R₃Are all propionic anhydride, R₄Is composed ofAs shown at 1c in fig. 1.

Example 4 provides AICAR analogs wherein R₁is-OH, R₂And R₃Form a ring to form a structureR₄Is composed ofAs shown at 1d in fig. 1. Example 5 provides AICAR analogs wherein R₁is-OH, R₂And R₃Form a ring to form a structureR₄Is composed ofAs shown at 1e in fig. 1.

Example 6 provides AICAR analogs wherein R₁is-OH, R₂And R₃Form a ring to form a structureR₄Is composed ofAs shown at 1f in fig. 1.

Examples 7 to 11

Examples 7 to 11 of this group each provide an adenosine analogue having a general molecular structure:

example 7 provides adenosine analogues in which R₁is-OH, R₂And R₃Form a ring, R₄Is composed ofAs shown at 2a in figure 2.

Example 8 provides adenosine analogues in which R₁、R₂And R₃Are all acetic anhydride, R₄Is composed ofAs shown at 2b in fig. 3.

Example 9 provides adenosine analogs in which R₁、R₂And R₃Are all propionic anhydride, R₄Is composed ofAs shown at 2c in fig. 3.

Example 10 provides adenosine analogs in which R₁、R₂And R₃Are each butyric anhydride, R₄Is composed ofAs shown at 2d in fig. 2.

Example 11 provides adenosine analogues in which R₁、R₂And R₃Are each isobutyric anhydride, R₄Is composed ofAs shown at 2e in figure 3.

Examples 12 to 17

Examples 12 to 17 in this group each provide an adenosine analogue having a general molecular structure:

example 12 provides the adenosine analogues in which R₁is-OH, R₂And R₃Form a ring to form a structureR₄Is composed ofAs shown at 2f in fig. 3.

Example 13 provides adenosine analogues in which R₁is-OH, R₂And R₃Form a ring to form a structureR₄Is composed ofAs shown at 2g in fig. 3.

Example 14 provides adenosine analogues in which R₁is-OH, R₂And R₃Form a ring to form a structureR₄Is composed ofAs shown at 2h in fig. 4.

Example 15 provides adenosine analogues in which R₁is-OH, R₂And R₃Form a ring to form a structureR₄Is composed ofAs shown at 2i in fig. 4.

Example 16 adenosine analogues in which R₁is-OH, R₂And R₃Form a ring to form a structureR₄Is composed ofAs shown at 2j in fig. 5.

Example 17 adenosine analogues in which R₁is-OH, R₂And R₃Form a ring to form a structureR₄Is composed ofAs shown at 2k in fig. 5.

Example 18

This example illustrates the preparation of example 1 by dissolving p-toluene sulfonic acid (31.25 g, 0.164mol triethyl orthoformate (39 ml, 0.234mol), AICAR (10g, 0.037mol) in acetone (1.5L) to give a mixture, stirring the resulting mixture at room temperature (20-25 deg.C) for 12h, then neutralizing with a saturated solution of carbonic acid, vacuum concentrating, cooling to crystallize, and filtering to give 11.4g white solid.

Example 19

This example shows the preparation of example 2, adding 10mL of anhydrous pyridine to AICAR (1g,0.0037mol), adding 1.12mL of acetic anhydride, stirring for 6h, detecting by thin layer chromatography that the reaction is complete, adding ice-cold anhydrous ethanol, stopping the reaction, vacuum concentrating to obtain a pale yellow oil, and performing column chromatography (50% -100%, ethyl acetate/petroleum ether); 1.32g of a white solid was obtained.

Example 20

This example shows the preparation of example 3, adding 10mL of anhydrous pyridine to adenosine (1g,0.0037mol), adding 1.94mL of propionic anhydride, stirring for 6h, detecting the reaction by thin layer chromatography, adding ice-cold anhydrous ethanol, and stopping the reaction. Vacuum-pumping and concentrating to obtain light yellow oily matter, and making column chromatographic separation (30% -100%, ethyl acetate/petroleum ether) to obtain white solid.

Example 21

This example shows the preparation of example 4 by adding 2mL of ultra dry tetrahydrofuran, anhydrous zinc chloride (3.5g, 0.257mol), and 4mL of 3' -methoxybenzaldehyde to adenosine (1g,0.0037mol), stirring at room temperature for 24h, detecting by thin layer chromatography that the reaction is complete, and stopping the reaction. 20mL of ethyl acetate was added thereto, and 6mL of a saturated solution of sodium hydrogencarbonate was added dropwise and then filtered. The organic phase in the filtrate was extracted with 30mL × 3 of ethyl acetate and washed with 100mL of water. Vacuum-pumping and concentrating to obtain light yellow oily matter, and making column chromatographic separation (50% -100%, ethyl acetate/petroleum ether); 1.249g of a white solid was obtained.

Example 22

This example illustrates the preparation of example 5 by adding 2mL of extra dry tetrahydrofuran to adenosine (1g,0.0037mol), adding anhydrous zinc chloride (3.5g, 0.257mol), 4mL3' -fluorobenzaldehyde, stirring at room temperature for 24h, detecting by thin layer chromatography that the reaction is complete, stopping the reaction, adding 20mL ethyl acetate, adding 6mL saturated sodium bicarbonate solution dropwise, filtering, extracting the organic phase from the filtrate with 30mL x 3 ethyl acetate, and washing the organic phase with 100mL water. Vacuum-pumping and concentrating to obtain light yellow oily matter, and making column chromatographic separation (50% -100%, ethyl acetate/petroleum ether); 0.53g of a white solid was obtained.

Example 23

This example shows the preparation of example 6, adding 2mL of ultra dry tetrahydrofuran, anhydrous zinc chloride (3.5g, 0.257mol), 4mL3' -chlorobenzaldehyde to adenosine (1g,0.0037mol), stirring at room temperature for 24h, detecting by thin layer chromatography that the reaction is complete, and stopping the reaction. 20mL of ethyl acetate was added thereto, and 6mL of a saturated solution of sodium hydrogencarbonate was added dropwise and then filtered. The organic phase in the filtrate was extracted with 30mL × 3 of ethyl acetate and washed with 100mL of water. Vacuum-pumping and concentrating to obtain light yellow oily matter, and making column chromatographic separation (50% -100%, ethyl acetate/petroleum ether); 0.21g of a white solid was obtained.

Example 24

This example shows the preparation of example 8, adding 10mL of anhydrous pyridine to adenosine (1g,0.0037mol), adding 1.12mL of acetic anhydride, stirring for 16h, detecting the reaction by thin layer chromatography, adding ice-cold anhydrous ethanol, and stopping the reaction. Vacuum-pumping and concentrating to obtain light yellow oily matter, and making column chromatographic separation (50% -100%, ethyl acetate/petroleum ether); 1.41g of a white solid was obtained.

Example 25

This example shows the preparation of example 9 by adding 10mL of anhydrous pyridine to adenosine (1g,0.0037mol), adding 1.53mL of propionic anhydride, stirring for 6h, detecting the reaction by thin layer chromatography, adding ice-cold anhydrous ethanol, and stopping the reaction. Vacuum-pumping and concentrating to obtain light yellow oily matter, and making column chromatographic separation (50% -100%, ethyl acetate/petroleum ether); 1.46g of a white solid was obtained.

Example 26

This example shows the preparation of example 10 by adding 10mL of anhydrous pyridine to adenosine (1g,0.0037mol), adding 1.94mL of butyric anhydride, stirring for 6h, detecting the reaction by thin layer chromatography, adding ice-cold anhydrous ethanol, and stopping the reaction. Vacuum-pumping and concentrating to obtain light yellow oily matter, and making column chromatographic separation (30% -100%, ethyl acetate/petroleum ether); 1.46g of a white solid was obtained.

Example 27

This example shows the preparation of example 11 by adding 10mL of anhydrous pyridine to adenosine (1g,0.0037mol), adding 1.96mL of isobutyric anhydride, stirring for 6h, detecting by thin layer chromatography that the reaction is complete, adding ice-cold anhydrous ethanol, and stopping the reaction. Vacuum-pumping and concentrating to obtain light yellow oily matter, and making column chromatographic separation (30% -100%, ethyl acetate/petroleum ether); 1.43g of a white solid was obtained.

Example 28

This example shows the preparation of example 12, adding 2mL of ultra dry tetrahydrofuran, anhydrous zinc chloride (3.5g, 0.257mol), 4mL of 3' -methoxybenzaldehyde to adenosine (1g,0.0037mol), stirring at room temperature for 24h, detecting the completion of the reaction by thin layer chromatography, and stopping the reaction. 20mL of ethyl acetate was added thereto, and 6mL of a saturated solution of sodium hydrogencarbonate was added dropwise and then filtered. The organic phase in the filtrate was extracted with 30mL × 3 of ethyl acetate and washed with 100mL of water. Vacuum-pumping and concentrating to obtain light yellow oily substance, and performing column chromatography (50% -100%, ethyl acetate/petroleum ether); 1.157g of a white solid were obtained.

Example 29

This example illustrates the preparation of example 13, adding 2mL of extra dry tetrahydrofuran to adenosine (1g,0.0037mol), adding anhydrous zinc chloride (3.5g, 0.257mol), 4mL3' -methylbenzaldehyde. After stirring for 24h at normal temperature, the reaction is detected to be complete by thin layer chromatography, and the reaction is stopped. 20mL of ethyl acetate was added thereto, and 6mL of a saturated solution of sodium hydrogencarbonate was added dropwise and then filtered. The organic phase in the filtrate was extracted with 30mL × 3 of ethyl acetate and washed with 100mL of water. Vacuum-pumping and concentrating to obtain light yellow oily substance, and performing column chromatography (50% -100%, ethyl acetate/petroleum ether); 1.378g of a white solid were obtained.

Example 30

This example shows the preparation of example 14, adding 2mL of ultra dry tetrahydrofuran, anhydrous zinc chloride (3.5g, 0.257mol), 4mL of p-tolualdehyde to adenosine (1g,0.0037mol), stirring at room temperature for 24h, detecting the completion of the reaction by thin layer chromatography, and stopping the reaction. Ethyl acetate (20 mL) was added, a saturated solution of sodium hydrogencarbonate (6 mL) was added dropwise, and the mixture was filtered, and the organic phase in the filtrate was extracted with ethyl acetate (30 mL. times.3), and then washed with 100mL of water. Vacuum-pumping and concentrating to obtain light yellow oily substance, and performing column chromatography (50% -100%, ethyl acetate/petroleum ether); 0.950g of a white solid was obtained.

Example 31

This example shows the preparation of example 15, adding 2mL of ultra dry tetrahydrofuran, anhydrous zinc chloride (3.5g, 0.257mol), 4mL3' -fluorobenzaldehyde to adenosine (1g,0.0037mol), stirring at room temperature for 24h, detecting by thin layer chromatography that the reaction is complete, and stopping the reaction. 20mL of ethyl acetate was added thereto, and 6mL of a saturated solution of sodium hydrogencarbonate was added dropwise and then filtered. The organic phase in the filtrate was extracted with 30mL × 3 of ethyl acetate and washed with 100mL of water. Vacuum-pumping and concentrating to obtain light yellow oily substance, and performing column chromatography (50% -100%, ethyl acetate/petroleum ether); 1.249g of a white solid was obtained.

Example 32

This example shows the preparation of example 16, adding 2mL of ultra dry tetrahydrofuran, anhydrous zinc chloride (3.5g, 0.257mol), 4mL3' -chlorobenzaldehyde to adenosine (1g,0.0037mol), stirring at room temperature for 24h, detecting the completion of the reaction by thin layer chromatography, and stopping the reaction. 20mL of ethyl acetate was added thereto, and 6mL of a saturated solution of sodium hydrogencarbonate was added dropwise and then filtered. The organic phase in the filtrate was extracted with 30mL × 3 of ethyl acetate and washed with 100mL of water. Vacuum-pumping and concentrating to obtain light yellow oily substance, and performing column chromatography (50% -100%, ethyl acetate/petroleum ether); 1.145g of a white solid was obtained.

Example 33

This example illustrates the preparation of example 17 by adding 2mL of extra dry THF to adenosine (1g,0.0037mol), adding anhydrous zinc chloride (3.5g, 0.257mol), 4mL3' -bromobenzaldehyde, stirring at room temperature for 24h, detecting by thin layer chromatography that the reaction is complete, and stopping the reaction. 20mL of ethyl acetate was added thereto, and 6mL of a saturated solution of sodium hydrogencarbonate was added dropwise and then filtered. The organic phase in the filtrate was extracted with 30mL × 3 of ethyl acetate and washed with 100mL of water. Vacuum-pumping and concentrating to obtain light yellow oily matter, and making column chromatographic separation (50% -100%, ethyl acetate/petroleum ether); 0.981g of a white solid was obtained.

Experimental example 1 running wheel test

In this experimental example, running wheel experiments for the sugar ring modified nucleoside analogs provided in the above examples were performed to characterize, and seven nucleoside analogs were selected from the existing nucleoside analogs for comparison, wherein CAS numbers corresponding to the seven selected nucleoside analogs are shown in the following table.

TABLE 1 seven existing sugar ring-modified nucleoside analogs selected in Experimental example 1

Numbering	CAS number
		2l	958-09-8
2m	168427-74-5
		2n	146-78-1
2o	10414-81-0
		2p	2140-79-6
2q	14365-44-7
		2r	892-48-8

1.1 against untrained mice

Blank untrained mice are adopted to simulate common people, 24 groups of experimental examples are set, the compounds provided in examples 1-17 and the selected seven existing nucleoside analogs are injected into the abdominal cavity respectively, 0.0196mmol/kg/d is set, meanwhile, a blank group injected with 0.1mL/d of physiological saline is set, and a red bull group injected with 0.5mL/d of the stomach-perfused red bull functional beverage is set. After 30 minutes, adjusting the rotating wheel type fatigue instrument of the mouse to 16r/min, and performing adaptive movement for 10 min; then adjusting the mouse wheel type fatigue instrument to 18r/min, and performing adaptive movement for 5 min; adjusting the rotating wheel type fatigue instrument of the mouse to 20r/min, and performing adaptive movement for 5 min; the wheel type fatigue instrument of the mice is adjusted to 22r/min, adaptive movement is carried out for 10min, and finally, the wheel type fatigue instrument of the mice is adjusted to 24r/min to measure the exhaustion distance of each group of mice at regular time, and the result is shown in figure 6.

As can be seen from fig. 6, the red cattle showed a certain physical fitness enhancing effect; the synthesized compound is administrated once, most of the compounds show clear physical ability improving effect, and half of the compounds are better than those of a red cattle control group; the optimal compound 2a is 17 times that of the blank group and 3.4 times that of the positive red cattle group.

1.2 for training mice

Firstly, training a mouse, adjusting a mouse wheel type fatigue instrument to 16r/min and performing adaptive movement for 10min after 30 minutes by adopting the same experimental group setting mode as that of the experimental example 1.1; then adjusting the mouse wheel type fatigue instrument to 18r/min, and performing adaptive movement for 5 min; adjusting the rotating wheel type fatigue instrument of the mouse to 20r/min, and performing adaptive movement for 5 min; the wheel type fatigue instrument of the mouse is adjusted to 22r/min, the mouse is exercised for 30 minutes, and the training is carried out for 5 days.

On the sixth day, the trained mouse wheel type fatigue instrument is adjusted to 16r/min, and adaptive movement is carried out for 10 min; then adjusting the mouse wheel type fatigue instrument to 18r/min, and performing adaptive movement for 5 min; adjusting the rotating wheel type fatigue instrument of the mouse to 20r/min, and performing adaptive movement for 5 min; the rotating wheel type fatigue instrument of the mouse is adjusted to 22r/min, and adaptive movement is carried out for 10 min. Finally, the pressure was adjusted to 24r/min to start timing and the exhaustion distance was measured, and the results are shown in FIG. 7.

As can be seen from fig. 7, the red cattle had limited effect on enhancing physical performance of mice trained for one week; the synthesized compound is administrated once, most of the compounds show clear physical ability improving effect, and half of the compounds are better than those of a red cattle control group; the optimal compound 2 is 3.7 times of that of the blank group and 1.7 times of that of the positive red cattle group.

1.3 Pre-and post-training comparison of mice

Taking the intersection of the compound preference of the blank mouse and the mouse moving one week in the experimental examples 1.1 and 1.2, 2a,2i,2m,2r are selected, as shown in fig. 8, it can be seen that the training can improve the exhaustion distance; the exhaustion distance of the blank group after one week of training is far lower than that of the untrained mice of the blank group with single administration, in other words, the amplification effect obtained by the single administration is far better than that of the training; preferred compounds can greatly increase the distance to exhaustion for the blank; the compound 2a can improve the effect by a single administration, which is higher than the known literature data.

Experimental example 2 swimming test

2.1 against untrained mice

The untrained mice are adopted to simulate the general population, the compound obtained by optimization in experimental example 1.3 is adopted to set the experimental group, after the mice are injected with the corresponding dose in the abdominal cavity for 30 minutes, the water temperature is 25 ℃, the water depth is 30 cm, the swimming test exhaustion time starts to bear the weight (6% of the weight, and the tin wire is loaded at the tail), the calculated exhaustion is realized without floating out of the water surface within 7s, and the result is shown in figure 9. The preferred compounds were further tested in the swimming model and all performed better than the blank.

2.2 for training mice

After the mice are trained for one week by adopting the method of the experimental example 2.1, the test time of swimming test is continuously 5 days, the water temperature is 25 ℃, the water depth is 30 cm, no load is applied, the swimming time is usually 50min, after the corresponding dose is injected into the abdominal cavity for 30 minutes on the 6 th day, the water temperature is 25 ℃, the water depth is 30 cm, the load begins to be applied (6% of the weight, the tin wire load is at the tail part), the result is shown in figure 10, and the preferable compound is further verified in the swimming model, and the effect is better than that of the blank group.

Experimental example 3 high jump test

The compound preferably obtained in example 1.3 was used to set the experimental group and the maximum average jump height was measured using the jump model. The water is heated at the outer edge of the bottom of the transparent water tank, the temperature of the hot water is 65 ℃, the temperature of the post surface is maintained at 50 ℃, the mouse jumps due to scorching of feet, the average maximum height is tested at the moment, and the result is shown in figure 11, so that the preferable compound can not only improve the movement endurance of the mouse, but also improve the explosive power of the mouse.

Experimental example 4

AutoDockVina is open source molecule docking software developed by Scripps laboratories, local optimization is performed by a quasi-newton method, a scoring function combines the advantages of scoring functions based on experience and knowledge, and affinity of receptor-ligand complexes is evaluated by calculating equivalent comprehensive scores such as space effect, repulsion, hydrogen bond, hydrophobic interaction and molecule flexibility of the receptor-ligand complexes, and the affinity is used as an important index for measuring whether ligands can be effectively combined with receptor molecules.

The invention takes AMPK protein (PDB Code: 4CFF) obtained from PDB database as a receptor, and the nucleoside analogue and the AMPK receptor protein provided by the embodiment are subjected to molecular docking analysis. The results are shown in table 2, with target compound binding energies scored as follows:

TABLE 2 binding energy of nucleoside derivatives to AMPK receptor protein

	AICAR	1a	1b	1c	1d	1e	1f
								Binding energy	-5.8	-7.8	-7.6	-7.5	-7.4	-7.4	-7.3
	Adenosine (I)	2a	2b	2c	2d	2e	2f
								Binding energy	-6.2	-8.9	-7.3	-7.3	-7.1	-6.8	-6.4
		2g	2h	2i	2j	2k	2l
										-7.5	-7.6	-8.6	-8.1	-8.2	-8.0
Binding energy	2m	2n	2o	2p	2q	2r
									-8.7	-8.3	-8.1	-8.4	-7.9	-8.8

As can be seen from table 1, the binding energy of the nucleoside analogues provided by the present invention to AMPK receptor protein is lower than that of AICAR and adenosine, demonstrating that the binding energy of selected compounds to AMPK receptor protein is better than that of AICAR and adenosine, especially 2a, and that docking with AMPK is the highest and far better than other molecules, indicating that it may be optimal for AMPK agonistic activity and may have potential energy enhancing effects.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.