WO2022099578A1 - Application of quinazoline derivative in preparation of drugs for preventing and/or treating gastrointestinal diseases - Google Patents

Abstract

An application of a quinazoline derivative in preparation of drugs for preventing or treating gastrointestinal diseases. The structural formula of the quinazoline derivative is shown in formula (I). Experimental results show that the quinazoline derivative has a significant therapeutic effect on peptic ulcer disease and inflammatory bowel disease, has the activity 20-80 times higher than existing clinical drugs, and can effectively improve mucosal integrity, promote tissue morphology improvement, and recover body weight of animals; moreover, in this field, the quinazoline derivative is mainly used for treating cardiovascular and cerebrovascular diseases, relating to new uses for old drugs; the quinazoline derivative has a good clinical application prospect in clinic.

Description

Application of quinazoline derivatives in the preparation of drugs for preventing and/or treating gastrointestinal diseases technical field

The invention relates to the technical field of medicine, in particular to the application of quinazoline derivatives in the preparation of medicines for preventing and/or treating digestive gastrointestinal diseases.

Background technique

Gastrointestinal diseases cover diseases of the esophagus, stomach, small intestine, colon and rectum. Common main symptoms include rhythm, periodic epigastric pain, diarrhea, hungry abdominal pain, acid regurgitation, fever, black stool, gastrointestinal bleeding and intestinal obstruction, etc. Gastrointestinal diseases are among the most common diseases in humans, the most common of which include dysphagia, gastric ulcer, peptic ulcer, gastroparesis, delayed gastric emptying, irritable bowel syndrome (IBS) and inflammatory bowel disease (IBD). Peptic ulcers mainly include gastric ulcers, duodenal ulcers and complex ulcers. At present, it is believed that the formation factors of gastric ulcers mainly focus on the weakening of the gastric mucosal barrier and the increase of gastrin secretion, while the formation of duodenal ulcers Factors are more focused on the increase in the total volume of parietal cells. In addition, exogenous factors such as excessive drinking, irregular eating, long-term mental stress and long-term use of non-steroidal anti-inflammatory drugs (such as aspirin), glucocorticoids, clopidogrel and other drugs are related to the formation of peptic ulcer disease. Inflammatory bowel disease can be caused by bacteria, fungi, viruses, parasites, protozoa and other organisms, and can also be caused by allergic reactions and physical and chemical factors. According to different etiologies, it can be divided into specific inflammatory lesions and non-specific inflammatory lesions. The former refers to infectious colitis, ischemic colitis and pseudomembranous colitis, and the latter mainly includes ulcerative colitis. , UC) and colonic Crohn's disease (Crohn's Disease, CD). UC is an inflammatory disease (Inflammatory bowel disease, IBD). As a common disease in gastroenterology, it can cause long-term inflammation and ulcers in the digestive tract. It is mainly characterized by chronic recurrent intestinal inflammation and intestinal epithelial cell damage. 20-30 years old, the main clinical symptoms are diarrhea, abdominal pain, bloody stool and intestinal obstruction. Many studies have shown that the pathogenesis of UC is related to factors such as immune factors, inflammation, environmental genetics, stress and infection. Gastritis consists of chronic gastritis and acute gastritis.

At present, the main types of western medicines for the treatment of gastrointestinal diseases include gastrointestinal motility drugs, antispasmodics, antiemetics, peptic ulcer drugs, gastric mucosal protective agents, digestive aids, probiotics, etc. Among them, the drugs for the treatment of peptic ulcer mainly include proton pump inhibitors, H2-receptor antagonists, bismuth preparations and prostaglandins, etc., which mainly neutralize gastric acid through raw materials containing calcium, although they can improve and relieve symptoms, but It is difficult to completely cure, the curative effect is unsatisfactory, and it is easy to recur. Inflammatory bowel disease is listed as one of the modern intractable diseases by the World Health Organization, and the incidence rate is on the rise in the world. The pathogenesis of colitis has not been fully elucidated. Commonly used drugs for clinical treatment include aminosalicylic acid preparations, sugar Corticosteroids and immunosuppressants, etc., short-term use can control the symptoms of colitis, but the cure rate is extremely low, long-term use can induce a variety of adverse reactions, and withdrawal can cause recurrence and other problems, and severe cases can cause cancer. Therefore, the development of The development of a new drug for the treatment of gastrointestinal diseases is a technical problem that needs to be solved urgently.

In the prior art, the quinazoline derivatives of formula (I) are mainly used for the treatment of cardiovascular and cerebrovascular diseases (CN2013106653967), cerebrovascular diseases include cerebral thrombosis, cerebral ischemia, cerebral infarction, etc., and cardiovascular diseases include myocardial Infarction, myocardial ischemia, myocardial injury, coronary heart disease, angina pectoris, heart failure, etc. It is also used for the treatment and/or prevention of sepsis and its complications, as well as for the use of activating new targets (EP2924037A1), however, there is currently no document that discloses the use of the quinazoline derivatives for the treatment of the gastrointestinal tract disease, and there is no technical suggestion to suggest to those skilled in the art that the quinazoline derivatives can be used for the treatment of gastrointestinal diseases. During the research process of the present invention, it was unexpectedly found that terazosin has a significant curative effect in the treatment of gastrointestinal diseases, and terazosin is a classic quinazoline derivative. Therefore, the inventor deduced from this that the formula (I) The quinazoline derivatives described have significant curative effects in the treatment of gastrointestinal diseases, can be used to prepare medicines for the treatment of gastrointestinal diseases, and provide a new use for quinazoline derivatives. In order to verify the above inference, the invention People also chose prazosin among quinazoline derivatives for verification experiments. The experimental results showed that prazosin also has significant curative effect in the treatment of gastrointestinal diseases, confirming the above inference, and prazosin and terazole The substituents of the oxazine are different, but the pharmacodynamics does not change significantly. Therefore, the quinazoline derivatives have the effect of treating gastrointestinal diseases.

SUMMARY OF THE INVENTION

In view of the above technical problems, the present invention discloses the application of quinazoline derivatives in the preparation of medicines for preventing or treating gastrointestinal diseases. The structural formula of the quinazoline derivatives is shown in formula (I):

or a tautomer, meso, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof; wherein:

R ₁ and R ₂ are each independently selected from H, NH ₂ , OH, C _1-6 alkyl-, C _1-6 alkoxy-C _1-6 alkyl-, C _2-6 alkenyl, C _{2 -6} alkynyl-, C _1-6 alkoxy-, C _1-6 alkylacyl-, aryl acyl-, C _6-10 aryl- or C _5-6 cycloalkyl-, the alkyl optionally substituted with _1-3 substituents selected from; hydroxy or halogen; or R1 and R2 together with the nitrogen atom to which they are attached form a 5- or ₆ -membered ring;

R ₃ and R ₄ are each independently selected from H, halogen, C _1-6 alkyl-, halogenated C _1-6 alkyl-, C _2-6 alkenyl-, C _2-6 alkynyl-, CN, NO ₂ , NH ₂ , OH, C _1-6 alkoxy-, C _1-6 alkoxy-C _1-6 alkoxy-, C _1-6 alkanoyloxy-, C _1-6 alkanoyl Amino-, _Ci -6alkanoylamino-, arylacylamino-, saturated or unsaturated 5- or 6-membered carbocyclyl or heterocyclyl, saturated or unsaturated 5- or 6-membered carbocycle Alkyloxy or heterocyclyloxy or _C1-6 alkylacyl; or _R3 and R4 together with the ring atoms to which they are attached form a 5- or ₆ -carbocyclic or heterocyclic ring;

R ₅ and R ₆ are each independently selected from H, halogen, CN, NO ₂ , NH ₂ , OH, C _1-6 alkyl-, C _1-6 alkoxy-C _1-6 alkoxy-, C _1-6 alkanoylamino, halogenated C _1-6 alkyl-, C _2-6 alkenyl-, C _2-6 alkynyl-, C _1-6 alkanoylamino-, C _1-6 alkanoylamino- , arylacylamino-, saturated or unsaturated 5- or 6-membered carbocyclyl or heterocyclyl, saturated or unsaturated 5- or 6-membered carbocyclyloxy or heterocyclyloxy or C _1-6 alkyl acyl;

R ₇ is selected from H, OH, NH ₂ , C _1-6 alkyl-, halogen-substituted C _1-6 alkyl-, alkoxy-substituted C _1-6 alkyl-, C _3-6 cycloalkyl- , spirocycloalkyl-, bridged cycloalkyl-, phenyl-, heterocyclyl, wherein the phenyl or heterocyclyl may be unsubstituted or one or more groups selected from the following Substituted, these groups include: C _1-6 alkyl-, C _1-6 alkoxy-, halogen-substituted C _1-6 alkyl-, nitro-, amino-, nitrile-, halogen-, C _{1 -6} sulfide or its oxidation product sulfone and sulfoxide.

Preferably, the quinazoline derivative of general structural formula (I), or its tautomer, meso, racemate, enantiomer, diastereomer or mixture thereof form, or a pharmaceutically acceptable salt thereof; the R ₇ is selected from C _1-6 alkyl-, halogen-substituted C _1-6 alkyl-, alkoxy-substituted C _1-6 alkyl-, C _{3 -6} cycloalkyl-, spirocycloalkyl-, bridged cycloalkyl-, phenyl-, heterocyclyl, wherein the phenyl or heterocyclyl may be unsubstituted or selected from the following One or more groups are substituted, these groups include: C _1-6 alkyl-, C _1-6 alkoxy-, halogen-substituted C _1-6 alkyl-, nitro-, amino-, nitrile- , halogen-, C _1-6 thioether or its oxidation product sulfone, sulfoxide.

Preferably, the quinazoline derivatives of the present invention include but are not limited to the following compounds:

Preferably, the quinazoline derivatives are prazosin and pharmaceutically acceptable salts thereof or terazosin and pharmaceutically acceptable salts thereof.

Preferably, the gastrointestinal disease is peptic ulcer.

Preferably, the peptic ulcer is one or more of gastric ulcer, duodenal ulcer, retrobulbar ulcer, pyloric ulcer, compound ulcer and kiss ulcer.

Preferably, the gastrointestinal disease is inflammatory bowel disease.

Preferably, the inflammatory bowel disease is one or more of ulcerative colitis and Crohn's disease.

Preferably, the quinazoline derivative and one or more pharmaceutically acceptable carriers make up the pharmaceutical composition.

Preferably, the dosage form of the pharmaceutical composition is injection, tablet, capsule, granule, and pill.

The beneficial effects of the present invention are: (1) The general structural formula of the quinazoline derivatives of the present invention is shown in formula (I), and the quinazoline derivatives of formula (I) are mainly used for the treatment of cardiovascular and cerebrovascular diseases , and the present invention is used for the treatment of gastrointestinal diseases, and has achieved good therapeutic effect, which belongs to the new use of old drugs. ② The present invention provides the application of the quinazoline derivatives in the preparation of drugs for preventing or treating gastrointestinal diseases, the curative effect is remarkable, the activity is 20-80 times higher than that of the existing clinical drugs, and it can effectively improve the integrity of the mucosa and promote the improvement of tissue morphology , recovery of animal body weight, can be popularized and applied clinically.

Description of drawings

Figure 1 The effect of terazosin on body weight difference and DAI score of different strains of UC mice;

Figure 2 The effect of terazosin on colon morphology and colon length in different strains of UC mice;

Figure 3 The effect of prazosin on body weight and DAI score of C57BL/6N colitis mice;

Fig. 4 The effect of prazosin on colon morphology and colon length in C57BL/6N colitis mice;

^### p<0.001vs.Control group; *p<0.05, **p<0.01 and ***p<0.001vs.DSS group.

Figure 5 The effect of terazosin and prazosin on gastric tissue morphology in mice with gastric ulcer;

Figure 6 Effects of terazosin and prazosin on gastric ulcer index, gastric ulcer area and gastric ulcer inhibition rate;

^### p<0.001 vs. Control group; **p<0.01 and ***p<0.001 vs. Ethanol group.

Detailed ways

The protection scope of the present invention will be described in detail below with reference to specific examples, but it should be noted that the protection scope of the present invention is not limited to the following examples, and a class of compounds that protect quinazoline derivatives in all peptic ulcer diseases and Therapeutic effects in inflammatory bowel disease, including different dosage forms, doses, combination drugs, etc. All technical solutions obtained by those skilled in the art through logical analysis, inference and experimentation in the prior art according to the idea of the present invention belong to the scope of the claimed protection of the present invention.

The C57BL/6N mouse described in the present invention is one of the most widely used strains in mice, and is also most commonly used as the female parent of transgenic or gene knockout mice in genetic engineering.

The KM mice of the present invention refer to Kunming mice.

In the present invention, the alkyl group refers to straight-chain and branched-chain saturated and unsaturated alkyl groups or aliphatic groups having a specific number of carbon atoms.

The alkoxy groups refer to saturated and unsaturated alkyl-O- groups having the specified number of carbon atoms.

The carboxylate-substituted alkyl group refers to a saturated and unsaturated alkyl-COOCH _X(X=0-3) -group having a specified number of carbon atoms.

The thioethers refer to saturated and unsaturated alkyl-S- groups having the indicated number of carbon atoms.

烷基基团。 The oxidation product sulfone refers to saturated and unsaturated carbon atoms with a specific number of carbon atoms.

alkyl group.

烷基基团。 Described oxidation product sulfoxide refers to saturated and unsaturated with specific number of carbon atoms.

alkyl group.

The "halogen" or "halo" refers to fluorine, chlorine, bromine or iodine as a substituent. When a halogen atom is used as a substituent, the number of substitutions on the same carbon atom can be one, two or three.

The aryl group includes phenyl, substituted phenyl (here, substituted phenyl includes one, two or three of the following groups: C _1-6 saturated and unsaturated alkyl, C _1-6 saturated and unsaturated alkoxy, phenoxy, nitrile, nitro, amino or halogen, etc.).

The term "heterocycle" as used in the present invention refers to stable 5- to 7-membered heterocycles, which may be saturated or unsaturated, and consist of carbon atoms and 1 to 4 optionally selected from N, O and S. It consists of 6 heteroatoms, in which nitrogen and sulfur heteroatoms can be selectively oxidized, and nitrogen heteroatoms can be selectively quaternized, preferably 6-membered heterocycles, such as pyridine, piperidine, pyrazine, piperazine, morpholine or thiomorpholine, etc.

The following examples of the present invention use sodium dextran sulfate to model colitis in mice.

In the following examples of the present invention, the indications of sulfasalazine enteric-coated tablets are (1) ulcerative colitis to treat mild to moderate ulcerative colitis; it can be used as an adjuvant therapy in severe ulcerative colitis. It can also be used for maintenance treatment of ulcerative colitis in remission; (2) Crohn's disease is used to treat active Crohn's disease, especially those patients with colon involvement; (3) rheumatoid arthritis is resistant to salicylic acid or other For rheumatoid arthritis and juvenile rheumatoid arthritis (polyarticular type) where the efficacy of non-steroidal anti-inflammatory drugs is not significant, sulfasalazine enteric-coated tablets are used as positive drugs in the first and second embodiments of the present invention .

Cimetidine in the following embodiments of the present invention, also known as cimetidine, is a histamine H2 receptor antagonist, mainly used to inhibit the secretion of gastric acid, can significantly inhibit the secretion of basal and nocturnal gastric acid, and can also inhibit the secretion of gastric acid. Gastric acid secretion caused by stimuli such as histamine, peptide gastrin, insulin and food, and reduce its acidity, prevent and protect corrosive gastritis caused by chemical stimulation Bleeding is also effective. The third and fourth embodiments of the present invention use cimetidine as a positive drug.

Embodiment 1. The therapeutic effect of terazosin on mice with different strains of colitis

1. Experimental animals, materials and sources

Experimental animals: 8-week-old SPF grade C57BL/6N male mice, Kunming (Kunming, KM) male mice and Balb/c male mice, weighing 18-22 g, without any drugs before the experiment, purchased from the Chinese Academy of Agricultural Sciences Lanzhou Veterinary Research Institute. The experimental animals were acclimatized and reared for a week in an environment with a temperature of 24-26°C and alternating daytime patterns of 12h/12h.

Drugs and reagents: Terazosin (Terazosin, TZ, HPLC≥98%), purchased from Aladdin Biotechnology Co., Ltd.; Sulfasalazine (Salazosulfapyridine, SASP) enteric-coated tablets purchased from Shanghai Xinyi Tianping Pharmaceutical Co., Ltd. ; Dextran Sulfate sodium (Dextran Sulfate sodium, DSS), MW: 40000, purchased from Aladdin Biotechnology Co., Ltd.

2. Experimental method

2.1 Preparation of ulcerative colitis model in C57BL/6N mice

40 C57BL/6N male mice were randomly divided into 5 groups, with 8 mice in each group. The groups and doses are shown in Table 1.

Table 1 Experimental grouping of C57BL/6N male mice and the dosage of each group

group Dosage Blank control group (Control) Distilled water for free drinking Blank control group+TZ group Free drinking of distilled water +4mg/kg/day DSS model control group 4%m/v DSS free drinking Positive control group (DSS+SASP) 4%m/v DSS free drink +80mg/kg/day DSS+TZ group 4%m/v DSS free drink +4mg/kg/day

4% DSS distilled water solution was prepared. DSS model control group mice freely drank DSS water solution for modeling, DSS+TZ group mice freely drank DSS water solution for modeling, and were also given 4 mg/kg/day terazosin; DSS+SASP group Mice freely drank DSS aqueous solution for modeling and were given 80 mg/kg/day of SASP; mice in blank control group drank freely distilled water, while mice in blank control group + TZ group drank freely distilled water and were given 4 mg/kg/day of terazole oxazine, all mice were fed with regular chow, and the model was continuously established for 7 days. On the seventh day, 2 hours after the last administration, the serum and colon tissue were separated for use.

2.2 Preparation of ulcerative colitis model in KM mice

Thirty KM male mice were randomly divided into 5 groups with 6 mice in each group. The groups and doses are shown in Table 2.

Table 2 Experimental grouping of KM male mice and the dosage of each group

group Dosage Blank control group (Control) Distilled water for free drinking Blank control group+TZ group Free drinking of distilled water +4mg/kg/day TZ DSS model control group 4%m/v DSS free drinking Positive control group (DSS+SASP) 4%m/v DSS free drink +80mg/kg/day SASP DSS+TZ group 4%m/v DSS free drink +4mg/kg/day TZ

4% DSS distilled water solution was prepared. DSS model control group mice freely drank DSS water solution for modeling, DSS+TZ group mice freely drank DSS water solution for modeling, and were also given 4 mg/kg/day terazosin; DSS+SASP group Mice freely drank DSS aqueous solution for modeling and were given 80 mg/kg/day of SASP; mice in blank control group drank freely distilled water, while mice in blank control group + TZ group drank freely distilled water and were given 4 mg/kg/day of terazole oxazine, all mice were fed with regular chow, and the model was continuously established for 7 days. On the seventh day, 2 hours after the last administration, the serum and colon tissue were separated for use.

2.3 Preparation of the Balb/c mouse ulcerative colitis model

Thirty Balb/c male mice were randomly divided into 5 groups, with 6 mice in each group. The groups and doses are shown in Table 3.

Table 3 Experimental groups of Balb/c male mice and the dosage of each group

group Dosage Blank control group (Control) Distilled water for free drinking Blank control group+TZ group Free drinking of distilled water +4mg/kg/day DSS model control group 4%m/v DSS free drinking Positive control group (DSS+SASP) 4%m/v DSS free drink +80mg/kg/day DSS+TZ group 4%m/v DSS free drink +4mg/kg/day

4% DSS distilled water solution was prepared. DSS model control group mice freely drank DSS water solution for modeling, DSS+TZ group mice freely drank DSS water solution for modeling, and were also given 4 mg/kg/day terazosin; DSS+SASP group Mice freely drank DSS aqueous solution for modeling and were given 80 mg/kg/day of SASP; mice in blank control group drank freely distilled water, while mice in blank control group + TZ group drank freely distilled water and were given 4 mg/kg/day of terazole oxazine, all mice were fed with regular chow, and the model was continuously established for 7 days. On the seventh day, 2 hours after the last administration, the serum and colon tissue were separated for use.

3. Detection of clinical indicators of ulcerative colitis

3.1 Weight loss situation

Calculated by the percentage (%) of body weight loss of each rat, no weight loss in the stomach is 1 point, 1%-5% weight loss is scored as 1 point, 6%-10% body weight loss is scored as 2 points, body weight loss is 11%-15% Scored as 3 points, weight loss greater than 15% was scored as 4 points.

3.2 Stool viscosity

Normal stool was scored as 0, loose stool as 2, and diarrhea as 4.

3.3 Blood in stool and occult blood

The normal stool score was recorded as 0 points, the occult blood bleeding was recorded as 1 point (judged according to the occult blood test results), and the bloody stool with the naked eye was recorded as 3 points.

The average of the three items of weight loss, stool consistency, and blood in the stool and occult blood is the DAI. The average of the above three items is DAI.

3.4 Measurement of colon length

After the mice were sacrificed, the colons of the mice were separated, and the ileum was cut from the junction of the ileum and the colon, and then at the colon near the anus. to the length of the anus, and photographed to record.

4. Data processing

The experimental data were statistically analyzed by SPSS23.0 software, and the data were expressed as (x±s), and the comparison between groups was performed by one-way analysis of variance (One-way ANOVA) and LSD-t method. p<0.05 was considered statistically significant.

5. Analysis of results

5.1 The effect of terazosin on body weight and disease activity index of different strains of colitis mice

During the experiment, the above-mentioned mice were monitored for their body weight, fecal viscosity, blood in the stool and occult blood, and SPSS 23.0 software was used to perform one-way ANOVA for data statistics.

Significant differences in body weight differences and DAI scores of mice in each group were expressed as *p<0.05, **p<0.01, and ***p<0.001. The body weight difference and DAI score detection results of each group of mice of different strains of UC mice are shown in Figure 1. It can be seen from the above data that compared with the blank control group, the blank control + TZ administration group showed no statistical difference in body weight index and DAI score index, indicating that terazosin has no toxic and side effects on normal mice; Compared with the DSS model group, the body weight of the DSS model group decreased significantly, and the DAI score showed a significant upward trend (###p<0.001), indicating that the DSS model was successfully prepared; Significant improvements in weight loss and DAI scores were observed (*p<0.05, **p<0.01 and ***p<0.001), and the efficacy of terazosin was stronger than that of the clinical drug sulfasalazine.

5.2 The effect of terazosin on colon morphology and colon length in different strains of colitis mice

After the mice in the above groups were sacrificed, their colon tissues were quickly removed, and their morphological changes were observed, and their lengths were measured at the same time, and statistical analysis was performed on the data of each group. Significant differences in colon length of mice in each group were expressed as *p<0.05, **p<0.01 and ***p<0.001.

The colon morphology and colon length monitoring results of each group of mice of different strains of UC mice are shown in Figure 2. Compared with the blank control group, there was no obvious change in the colon length of the mice in the blank control + TZ administration group, indicating that the quinazoline derivative terazosin had no toxic and side effects on normal mice; compared with the blank control group, the DSS model The colon length of the group was significantly shortened (###p<0.001), indicating that the DSS model was successfully prepared; compared with the DSS model control group, each administration group could significantly improve the colonic edema and shortening, and it was significant Differences (*p<0.05, **p<0.01 and ***p<0.001), and the efficacy of the quinazoline derivative terazosin was stronger than that of the clinical drug sulfasalazine.

The measurement of body weight difference, DAI score, colon morphology and colon length showed that the quinazoline derivative terazosin was effective on DSS-induced colitis mice of different strains, and the curative effect was significantly stronger than the clinical drug sulfasalazine Efficacy.

During the research process of the present invention, it was unexpectedly found that terazosin has a significant curative effect in the treatment of gastrointestinal diseases. The experimental results are shown in Example 1, and terazosin is a classic quinazoline derivative. It is inferred that the quinazoline derivatives of formula (I) also have effects in the treatment of gastrointestinal diseases, and can be used to prepare medicines for the treatment of gastrointestinal diseases, providing a new use for quinazoline derivatives. In order to verify the above inference, the inventor selected prazosin in the quinazoline derivatives for verification experiments, and set up Example 2 and Example 3.

Embodiment 2. The therapeutic effect of prazosin on C57BL/6N colitis mice

1. Experimental animals, materials and sources

Experimental animals: 8-week-old SPF grade C57BL/6N male mice, weighing 18-22 g, without any drugs before the experiment, were purchased from Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The experimental animals were acclimatized and raised for a week in an environment with a temperature of 24-26 °C and alternating daytime patterns of 12h/12h.

Drugs and reagents: Prazosin (Prazosin, PZ, HPLC≥98%), purchased from Aladdin Biotechnology Co., Ltd.; Sulfasalazine (Salazosulfapyridine, SASP) enteric-coated tablets were purchased from Shanghai Xinyi Tianping Pharmaceutical Co., Ltd.; Dextran Sulfate Sodium (DSS), MW: 40000, was purchased from Aladdin Biotechnology Co., Ltd.

2. Experimental method

2.1 Preparation of ulcerative colitis model in C57BL/6N mice

A total of 40 C57BL/6N male mice were randomly divided into 5 groups, with 8 mice in each group. The groups and doses are shown in Table 4.

Table 1 Experimental grouping of C57BL/6N male mice and the dosage of each group

group Dosage Blank control group (Control) Distilled water for free drinking Blank control group+PZ group Free drinking of distilled water + 2mg/kg/day PZ DSS model control group 4%m/v DSS free drinking Positive control group (DSS+SASP) 4%m/v DSS free drink +80mg/kg/day SASP DSS+PZ group 4% m/v DSS free drink + 2mg/kg/day PZ

4% DSS distilled water solution was prepared. DSS model control group mice freely drank DSS water solution for modeling, DSS+PZ group mice freely drank DSS water solution for modeling and were given 2 mg/kg/day prazosin; DSS+SASP group mice Rats in the blank control group were given free access to distilled water, while the mice in the blank control group and PZ group were given free access to distilled water and given 2 mg/kg/day of sulfasalazine. Zosin, all mice were kept on a regular chow diet, and the model was continuously established for 7 days. On the seventh day, 2 hours after the last administration, the serum and colon tissue were separated for use.

3. Detection of clinical indicators of ulcerative colitis

3.1 Weight loss situation

Calculated according to the percentage (%) of the body weight of each mouse, no weight loss is 1 point, 1%-5% weight loss is 1 point, 6%-10% weight loss is 2 points, 11%-15% body weight loss Scored as 3 points, weight loss greater than 15% was scored as 4 points.

3.2 Stool viscosity

Normal stool was scored as 0, loose stool as 2, and diarrhea as 4.

3.3 Blood in stool and occult blood

The normal stool score was recorded as 0 points, the occult blood bleeding was recorded as 1 point (judged according to the occult blood test results), and the bloody stool with the naked eye was recorded as 3 points.

The average of the three items of weight loss, stool consistency, and blood in the stool and occult blood is the DAI.

3.4 Measurement of colon length

After the mice were sacrificed, the colons of the mice were separated, and the ileum was cut from the junction of the ileum and the colon, and then at the colon near the anus. to the length of the anus, and photographed to record.

4. Data processing

The experimental data were statistically analyzed by SPSS23.0 software, and the data were expressed as (x±s), and the comparison between groups was performed by one-way analysis of variance (One-way ANOVA) and LSD-t method. p<0.05 was considered statistically significant.

5. Analysis of results

5.1 Effects of PZ compounds on body weight and disease activity index of C57BL/6N colitis mice

During the experiment, the above-mentioned mice were monitored for their body weight, fecal viscosity, blood in the stool and occult blood, and SPSS 23.0 software was used for one-way analysis of variance for data statistics.

Significant differences in body weight differences and DAI scores of mice in each group were expressed as *p<0.05, **p<0.01, and ***p<0.001. The body weight difference and DAI score detection results of each group of UC mice are shown in Figure 3. It can be seen from the above data that compared with the blank control group, the blank control + PZ administration group showed no statistical difference in body weight index and DAI score index, indicating that the quinazoline derivative prazosin has no toxic and side effects on normal mice. ; Compared with the blank control group, the body weight of the DSS model group decreased significantly, and the DAI score showed a significant upward trend (###p<0.001), indicating that the DSS model was successfully prepared; Both the drug groups could significantly improve body weight loss and DAI score (*p<0.05, **p<0.01 and ***p<0.001), and in the case of a 40-fold difference in the doses, the quinazoline derivatives pipette The efficacy of zosin is still stronger than that of the clinical drug sulfasalazine.

5.2 Effects of quinazoline derivative prazosin on colon morphology and colon length in different strains of colitis mice

After the mice in the above groups were sacrificed, their colon tissues were quickly removed, and their morphological changes were observed, and their lengths were measured at the same time, and statistical analysis was performed on the data of each group. Significant differences in colon length of mice in each group were expressed as *p<0.05, **p<0.01 and ***p<0.001.

The colon morphology and colon length monitoring results of each group of UC mice are shown in Figure 4 . Compared with the blank control group, the colon length of the mice in the blank control + PZ administration group did not change significantly, indicating that the quinazoline derivative prazosin had no toxic and side effects on normal mice; compared with the blank control group, the DSS model group The colon length was significantly shortened (###p<0.001), indicating that the DSS model was successfully prepared; compared with the DSS model control group, each administration group could significantly improve the colonic edema and shortening, and there were significant differences (*p<0.05, **p<0.01 and ***p<0.001), and the efficacy of the quinazoline derivative prazosin was stronger than that of the clinical drug sulfasalazine.

The measurement of body weight difference, DAI score, colon morphology and colon length showed that the quinazoline derivative prazosin was effective against DSS-induced colitis mice of different strains, and the curative effect was significantly stronger than that of clinical drug sulfasalazine. curative effect.

Embodiment three, the therapeutic effect of quinazoline derivatives terazosin and prazosin on KM gastric ulcer mice

1. Experimental animals, materials and sources

Experimental animals: 8-week-old SPF grade Kunming (Kunming, KM) male mice, weighing 18-22 g, without any drugs before the experiment, were purchased from Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The experimental animals were acclimatized and raised for a week in an environment with a temperature of 24-26 °C and alternating daytime patterns of 12h/12h.

Drugs and reagents: Terazosin (Terazosin, TZ, HPLC≥98%), purchased from Aladdin Biotechnology Co., Ltd.; Prazosin (Prazosin, PZ, HPLC≥98%), purchased from Aladdin Biotechnology Co., Ltd. ; Anhydrous ethanol, Tianjin Damao Chemical Reagent Company; Cimetidine Capsules (CIM) were purchased from Teyi Pharmaceutical Group Co., Ltd.

2. Preparation of KM mouse model of alcoholic gastric ulcer

50 KM male mice (20-25g) were raised in the experimental animal room of Gansu University of Traditional Chinese Medicine. After the mice were adaptively reared for one week, they were randomly divided into 5 groups with 10 mice in each group. As shown in Table 5;

Table 5 Experimental grouping of KM male mice and the dosage of each group

group Dosage Blank control group 0.3mL/gavage of normal saline Ethanol model group 0.3mL/only anhydrous ethanol by gavage Cimetidine positive group 80mg/kg/day cimetidine was administered by intragastric administration, and 0.3mL/only anhydrous ethanol was administered by intragastric administration after 2 hours

TZ administration group 1mg/kg/day TZ was administered by intragastric administration, and 0.3mL/piece was intragastrically administered with anhydrous ethanol after 2h PZ administration group 2mg/kg/day TZ was administered by intragastric administration, and 0.3mL/piece was intragastrically administered with anhydrous ethanol after 2h

The mice in the above groups were stimulated for 2 h to end the experiment, and the serum and gastric tissue were separated to determine various indicators.

3. Ulcer area, ulcer index and inhibition rate

Take out the stomach, cut it along the greater curvature of the stomach, rinse the contents, observe the gastric mucosal ulcer, measure the transverse diameter and longitudinal diameter of the ulcer with a ruler, and the product of the two is the ulcer area (mm ² ), as shown in formula 1 ; and then calculate the ulcer area of the entire gastric tissue, thereby calculating the ulcer inhibition rate (%), see formula 2. At the same time, the mean of the sum of the ulcer points in each group was used as the ulcer index (0 points for healing, 1 point for superficial mucosal erosion, 2 points for deep ulcers or transmural necrosis, and 3 points for perforated or penetrating ulcers). Minute)

Ulcer area (mm2) = maximum long diameter of ulcer * maximum wide diameter perpendicular to the maximum long diameter (1)

4. Data processing

The experimental data were statistically analyzed by SPSS23.0 software, and the data were expressed as (x±s), and the comparison between groups was performed by one-way analysis of variance (One-way ANOVA) and LSD-t method. p<0.05 was considered statistically significant.

5. Effects of quinazoline derivatives terazosin and prazosin on gastric tissue morphology, ulcer index, ulcer area and ulcer inhibition rate in gastric ulcer mice

After the mice in the above groups were sacrificed, the gastric tissue was quickly taken to observe the morphological changes and ulcer damage, and the ulcer area and ulcer index were statistically analyzed. The experimental results are shown in Figures 5 and 6. It can be seen from Figure 5 that the macroscopic morphology of the gastric tissue of the mice in the normal group was normal, and no obvious hemorrhagic lesions appeared, while the gastric tissue of the mice in the ethanol model group had obvious hemorrhagic lesions and the most serious ulcers, indicating that the ethanol gastric ulcer model was successfully prepared; Compared with the quinazoline derivative terazosin, quinazoline derivative prazosin and cimetidine, each administration group can significantly improve the degree of gastric mucosal damage, reduce the ulcer area, and quinazoline The ulcer inhibition rates of derivatives terazosin and quinazoline derivatives prazosin (81.57% and 77.35%) were higher than those of clinical drug cimetidine (CIM) (69.91%).

The above-mentioned ulcer area, ulcer index and inhibition rate, and gastric histomorphological measurements show that the quinazoline derivative terazosin and the quinazoline derivative prazosin are effective in preventing and treating gastric ulcer, and the curative effect is significantly stronger than clinical medication Efficacy of cimetidine.

The experimental results of Example 2 and Example 3 show that prazosin also has a significant curative effect in treating gastrointestinal diseases, confirming that quinazoline derivatives have the effect of treating gastrointestinal diseases, verifying the inventor's inference, and The substituents of prazosin and terazosin are different, but the efficacy shown in Example 3 does not change significantly. Therefore, the quinazoline derivatives have the effect of treating gastrointestinal diseases.

To sum up, the quinazoline derivatives terazosin and prazosin of the present invention, under different feeding conditions, were subjected to DAI score and colon morphometry in different strains of mice, and the results showed that quinazoline The derivative terazosin has the effect of treating ulcerative colitis in different genetic backgrounds, and the quinazoline derivative prazosin also has a certain therapeutic effect on ulcerative colitis. At the same time, under different feeding conditions, the ulcer index, inhibition rate and histomorphological examination of mice were carried out. The effect is that the quinazoline derivative prazosin of the present invention and the quinazoline derivative terazosin have the effect of preventing and treating ulcerative colitis and gastric ulcer, and the preventing and treating effect is significantly stronger than that of cimetidine. The quinazoline derivative of the invention has the effect of preventing and treating ulcerative colitis and gastric ulcer, and has a good clinical application prospect.

Claims (9)

The application of quinazoline derivatives in the preparation of drugs for preventing or treating gastrointestinal diseases, characterized in that the general structural formula of the quinazoline derivatives is shown in formula (I):

or a tautomer, meso, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof; wherein: R ₁ and R ₂ are each independently selected from H, NH ₂ , OH, C _1-6 alkyl-, C _1-6 alkoxy-C _1-6 alkyl-, C _2-6 alkenyl, C _{2 -6} alkynyl-, C _1-6 alkoxy-, C _1-6 alkylacyl-, aryl acyl-, C _6-10 aryl- or C _5-6 cycloalkyl-, the alkyl optionally substituted with _1-3 substituents selected from; hydroxy or halogen; or R1 and R2 together with the nitrogen atom to which they are attached form a 5- or ₆ -membered ring; R ₃ and R ₄ are each independently selected from H, halogen, C _1-6 alkyl-, halogenated C _1-6 alkyl-, C _2-6 alkenyl-, C _2-6 alkynyl-, CN, NO ₂ , NH ₂ , OH, C _1-6 alkoxy-, C _1-6 alkoxy-C _1-6 alkoxy-, C _1-6 alkanoyloxy-, C _1-6 alkanoyl Amino-, _Ci -6alkanoylamino-, arylacylamino-, saturated or unsaturated 5- or 6-membered carbocyclyl or heterocyclyl, saturated or unsaturated 5- or 6-membered carbocycle Alkyloxy or heterocyclyloxy or _C1-6 alkylacyl; or _R3 and R4 together with the ring atoms to which they are attached form a 5- or ₆ -carbocyclic or heterocyclic ring; R ₅ and R ₆ are each independently selected from H, halogen, CN, NO ₂ , NH ₂ , OH, C _1-6 alkyl-, C _1-6 alkoxy-C _1-6 alkoxy-, C _1-6 alkanoylamino, halogenated C _1-6 alkyl-, C _2-6 alkenyl-, C _2-6 alkynyl-, C _1-6 alkanoylamino-, C _1-6 alkanoylamino- , arylacylamino-, saturated or unsaturated 5- or 6-membered carbocyclyl or heterocyclyl, saturated or unsaturated 5- or 6-membered carbocyclyloxy or heterocyclyloxy or C _1-6 alkyl acyl; R ₇ is selected from H, OH, NH ₂ , C _1-6 alkyl-, halogen-substituted C _1-6 alkyl-, alkoxy-substituted C _1-6 alkyl-, C _3-6 cycloalkyl- , spirocycloalkyl-, bridged cycloalkyl-, phenyl-, heterocyclyl, wherein the phenyl or heterocyclyl may be unsubstituted or one or more groups selected from the following Substituted, these groups include: C _1-6 alkyl-, C _1-6 alkoxy-, halogen-substituted C _1-6 alkyl-, nitro-, amino-, nitrile-, halogen-, C _{1 -6} sulfide or its oxidation product sulfone and sulfoxide. The application according to claim 1, characterized in that, the quinazoline derivative of general structural formula (I), or its tautomer, meso, racemate, or enantiomer , diastereomer or its mixture form, or its pharmaceutically acceptable salt; said R ₇ is selected from C _1-6 alkyl-, halogen substituted C _1-6 alkyl-, alkoxy substituted C _1-6 alkyl-, C _3-6 cycloalkyl-, spirocycloalkyl-, bridged cycloalkyl-, phenyl-, heterocyclyl, wherein the phenyl or heterocyclyl can be unidentified is substituted or is substituted by one or more groups selected from the following groups, including: C _1-6 alkyl-, C _1-6 alkoxy-, halogen-substituted C _1-6 alkyl-, Nitro-, amino-, nitrile-, halogen-, C _1-6 thioether or its oxidation product sulfone, sulfoxide. The application according to claim 2, wherein the quinazoline derivative is prazosin and pharmaceutically acceptable salts thereof or terazosin and pharmaceutically acceptable salts thereof. The application according to any one of claims 1-3, wherein the gastrointestinal disease is peptic ulcer. The application according to claim 4, wherein the peptic ulcer is one or more of gastric ulcer, duodenal ulcer, retrobulbar ulcer, pyloric canal ulcer, compound ulcer and kiss ulcer . The use according to any one of claims 1-3, wherein the gastrointestinal disease is inflammatory bowel disease. The application according to claim 6, wherein the inflammatory bowel disease is one or more of ulcerative colitis and Crohn's disease. The use according to any one of claims 1-3, wherein the quinazoline derivative and one or more pharmaceutically acceptable carriers form a pharmaceutical composition. The application according to claim 8, wherein the pharmaceutical composition is in the form of injection, tablet, capsule, granule or pill.

Images