TW202247844A - Use of pyrrolopyrimidine compound - Google Patents

Abstract

A use of a pyrrolopyrimidine compound. The structure of the pyrrolopyrimidine compound is represented by formula I, and the pyrrolopyrimidine compound may be used to prepare a drug for treating and/or preventing inflammatory bowel disease.

Description

Uses of pyrrolopyrimidines

This application claims the priority of Chinese patent application 2021106018102 with a filing date of 2021/5/31 and Chinese patent application 2022102181538 with a filing date of 2022/2/28. This application cites the full text of the above-mentioned Chinese patent application.

The present invention relates to the use of a pyrrolopyrimidine compound.

Inflammatory bowel disease (Inflammatory Bowel Disease, IBD) is a chronic intestinal inflammatory disease. The composition and structure of intestinal flora in IBD patients have obvious changes, that is, intestinal flora dysbiosis. Intestinal barrier function plays an important regulatory role in maintaining the stable state of human intestinal function. Studies have shown that in IBD there is impaired intestinal barrier function and increased intestinal permeability. The dysbiosis of intestinal flora leads to the impairment of intestinal barrier function, thereby destroying the normal intestinal homeostasis function of the human body and aggravating the progression of IBD.

Inflammatory bowel disease mainly includes ulcerative colitis (Ulcerative Colitis, UC) and Crohn's disease (Crohn Disease, CD), which are closely related to autoimmune dysfunction. This disease is very common in Western countries. In recent years, the number of IBD cases in my country It has become a common disease of the digestive system and the main cause of chronic diarrhea. Clinically, patients with inflammatory bowel disease manifest as repeated abdominal pain, diarrhea, mucus and bloody stools, and even various systemic complications such as blurred vision, joint pain, and skin rashes. The disease can be improved after treatment, and can also be relieved by itself. However, most patients relapse and do not heal, and a considerable number of patients require surgical treatment due to complications. So far, its etiology and specific pathogenesis are still unclear, and there is no fundamental treatment plan for the disease clinically. The World Health Organization lists the disease as one of the modern refractory diseases.

Tofacitinib was approved by the U.S. Food and Drug Administration (FDA) for the treatment of autoimmune disease rheumatoid arthritis in November 2012, and in May 2018 for the treatment of moderate to severe For the treatment of ulcerative colitis, the clinical remission rate of high dose (10mg/kg) is about 40%. However, because it acts on the upstream cursor of the message, it can cause a variety of non-specific reactions, including hyperlipidemia, severe infection, malignant tumor and pulmonary embolism and other toxic and side effects, some of which can be life-threatening, and its toxicity is proportional to the dose. In view of this, the Safety Committee of the European Medicines Agency began to conduct a comprehensive evaluation of tofacitinib in 2019. At the same time, the US FDA also issued a black box warning (a rare safety warning) that tofacitinib may cause blood coagulation and death risks, and use it with caution Or discontinue the clinical dose of 10mg/kg, and compromise the clinical dose of 5mg/kg.

In view of the low clinical remission rate and serious adverse reactions of the latest drug tofacitinib. Therefore, the research and development of drugs that can safely and effectively prevent and treat inflammatory bowel disease has important guiding significance for the clinical treatment of inflammatory bowel disease.

The technical problem to be solved by the present invention is to provide a compound that has good preventive and/or therapeutic effects on inflammatory bowel disease. Therefore, the present invention provides a use of pyrrolopyrimidine compounds.

。 The present invention provides a use of a compound represented by formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicine for treating and/or preventing inflammatory bowel disease;

.

In some embodiments, the inflammatory bowel disease is ulcerative colitis.

In some embodiments, the inflammatory bowel disease is Crohn's disease.

In some embodiments, the inflammatory bowel disease has symptoms of blood in stool and/or diarrhea.

In some embodiments, the medicament for treating and/or preventing inflammatory bowel disease comprises a pharmaceutical adjuvant.

In some embodiments, the compound represented by formula I or a pharmaceutically acceptable salt thereof is one or the only active ingredient of the drug.

In some embodiments, a therapeutically effective amount of the compound represented by formula I or a pharmaceutically acceptable salt thereof may be administered to a subject in need. The mode of administration may be any suitable mode, such as oral administration.

The therapeutically effective amount administered may be 4-100 mg/kg/d, preferably 6-60 mg/kg/d, for example 6 mg/kg/d, 20 mg/kg/d or 60 mg/kg/d.

The administration frequency may be 1-3 times/day, for example, 1-2 times/day.

In the present invention, the compound represented by formula I or a pharmaceutically acceptable salt thereof can be administered to a subject by any suitable route, such as orally.

The present invention also provides a pharmaceutical composition for treating and/or preventing inflammatory bowel disease in a subject, which comprises: a compound represented by formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable adjuvant.

In some embodiments, the inflammatory bowel disease is ulcerative colitis or Crohn's disease.

The present invention also provides a method for treating and/or preventing inflammatory bowel disease in a subject, which comprises: administering to the subject a therapeutically or preventively effective amount of a compound represented by formula I or a pharmaceutically acceptable salt thereof .

In some embodiments, the inflammatory bowel disease is ulcerative colitis or Crohn's disease. Definition and Description

Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A specific term or phrase should not be considered indeterminate or unclear if it is not specifically defined, but should be understood according to its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding trade name or its active ingredient.

In the present invention, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms, which are suitable for use in contact with human and animal tissues within the scope of reliable medical judgment , without undue toxicity, irritation, allergic reaction or other problems or complications, commensurate with a reasonable benefit/risk ratio.

In the present invention, the term "pharmaceutically acceptable salt" may be a salt prepared from a compound of the present invention and a relatively non-toxic, pharmaceutically acceptable acid or base. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base, either neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts may include, but are not limited to: lithium salts, sodium salts, potassium salts, calcium salts, aluminum salts, magnesium salts, zinc salts, bismuth salts, ammonium salts, diethanolamine salts and the like. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compound with a sufficient amount of the acid, either neat solution or in a suitable inert solvent. The pharmaceutically acceptable acid may include inorganic acid, which may include but not limited to: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, sulfuric acid and the like. The pharmaceutically acceptable acid may include organic acids, which may include, but are not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, Diacid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acid citric acid, Oleic acid, tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentisic acid, butenedioic acid, gluconic acid, sugar acid, formic acid, ethanesulfonic acid, pamoic acid (ie 4,4'-methylene - bis(3-hydroxy-2-naphthoic acid)), amino acids (eg glutamic acid, arginine) and the like. When the compounds of the present invention contain relatively acidic and relatively basic functional groups, they can be converted into base addition salts or acid addition salts. See Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66: 1-19 (1977), or, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl and Camille G. Wermuth, ed., Wiley-VCH, 2002).

In the present invention, the medicament for treating and/or preventing inflammatory bowel disease can be in conventional dosage forms in the art, such as tablets, capsules, intravenous injections, intraperitoneal injections, inhalants, nebulizers, lyophilized agents, Patches, gels, sprays or suppositories, etc.

The term "pharmaceutical adjuvant" refers to the excipients and additives used in the production of medicines and formulation of prescriptions, which may be all substances contained in pharmaceutical preparations except active ingredients. Please refer to the Pharmacopoeia of the People's Republic of China (2020 Edition), Volume Four, or Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009 Sixth Edition).

The term "treatment" refers to therapeutic therapy. In relation to a specific condition, treatment means: (1) amelioration of one or more biological manifestations of the disease or condition, (2) interference with (a) one or more points in the biological cascade leading to or causing the condition or (b) the condition (3) ameliorate one or more symptoms, effects or side effects associated with the condition, or one or more symptoms, effects or side effects associated with the condition or its treatment, or (4) slow down the condition or the condition The development of one or more biological manifestations.

The term "prevention" refers to a reduction in the risk of acquiring or developing a disease or disorder.

The term "therapeutically effective amount" refers to an amount of a compound sufficient to effectively treat a disease or condition described herein when administered to a subject. A "therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, but can be adjusted as necessary by those skilled in the art.

The term "prophylactically effective amount" refers to an amount sufficient to prevent a disease or disorder, or to prevent one or more symptoms associated with a disease or disorder, or to prevent recurrence of a disease or disorder.

The term "subject" refers to any animal that is about to or has received the administration of the compound according to the embodiments of the present invention, preferably a mammal, and most preferably a human. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., with humans being most preferred.

On the basis of not violating common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.

The reagents and raw materials used in the present invention are all commercially available.

The positive progress effect of the present invention is that compound I or its pharmaceutically acceptable salt has good preventive or therapeutic effects on inflammatory bowel diseases such as ulcerative colitis and Crohn's disease.

detailed description

The present invention is further illustrated below by means of examples, but the present invention is not limited to the scope of the examples. For the experimental methods that do not specify specific conditions in the following examples, select according to conventional methods and conditions, or according to the product instructions. The solvent used in the present invention and the reference substance can all be commercially available.

Compound I was provided by Guangzhou Jiayue Pharmaceutical Technology Co., Ltd. and prepared according to Example 1 in WO2020244614A1;

Tofacitinib was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

Cyclosporin A, trade name Sandiming, was purchased from NOVARTIS.

Dextran sulfate sodium (Dextran sulfate sodium , DSS, MW=36,000-50,000): Cat. No.: 160110; Example 1 1. Experimental animals

Animal species and strain: C57BL/6 mice; female, weighing about 18-22 g, 6-8 weeks old; purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.; Table 1 Animal grouping and administration methods group group name number of animals compound Method of administration Route of administration Dosing frequency G1 normal control group ^a 8 solvent oral twice a day G2 Model control group ^a 12 solvent oral twice a day G3 Compound I group ^a 12 Compound Ⅰ oral twice a day G4 Cyclosporin A group ^b 12 cyclosporine A oral once a day G5 Tofacitinib group ^a 12 Tofacitinib oral twice a day G6 Compound I group ^a 12 Compound Ⅰ oral twice a day G7 Compound I group ^a 12 Compound Ⅰ oral twice a day a: The vehicle is 0.5% (w/v) hydroxypropylmethylcellulose (HPMC E5), 0.5% (w/v) polyvinylpyrrolidone (PVP K30) and 0.2% (w/v) sodium lauryl sulfate (SDS) mixed aqueous solution; b: The vehicle is physiological saline.

The mice were randomly divided into normal control group (G1), model control group (G2), compound I group (G3), positive drug cyclosporine A group (G4), tofacitinib group (G5), compound I group (G6) and compound I group (G7), animal grouping and administration methods are shown in Table 1 for details. Model control group (G2) and model administration group animals (G3-G7) were given drinking water containing 2% DSS (dextran sodium sulfate) continuously for 6 days to establish the model, and then replaced with normal drinking water, and samples were taken after 4 days . Mice in the normal control group were not subjected to model establishment and drank normal drinking water all the time. Samples were taken from the day the model was established to the end point. Animals in compound I group (G3) received oral administration of compound I (twice a day, 6 mg/kg/d), and animals in cyclosporine A group (G4) received oral administration of cyclosporine A Administration (once a day, 25mg/kg/d), animals in tofacitinib group (G5) received oral administration of tofacitinib (twice a day, 60mg/kg/d), animals in compound I group (G6 ) received oral administration of compound I (twice a day, 20mg/kg/d), compound I group animals (G7) received oral administration of compound I (twice a day, 60mg/kg/d), and animals in the control group were orally administered Give vehicle (vehicle is 0.5% (w/v) hydroxypropyl methylcellulose (HPMC E5), 0.5% (w/v) polyvinylpyrrolidone (PVP K30) and 0.2% (w/v) lauryl sulfate Sodium (SDS) mixed aqueous solution). All animals were dosed at a volume of 10 mL/kg. Animal body weight and clinical disease scores were recorded daily during the experiment. The body weight, diarrhea and hematochezia of mice in all groups were recorded every day. Daily scoring is done according to the scoring system below.

Weight change (0, <1%; 1, 1-5%; 2, 6-10%; 3, 11-20%;);

Bleeding score (0, occult blood negative; 1, occult blood positive; 2, trace blood visible to the naked eye in stool; 3, obvious rectal bleeding);

Stool score (0, normal; 1, mild stool; 2, very soft, wet stool; 3, watery diarrhea)

The scores of the three parts are added to obtain the daily disease index value DAI. 2. Histopathological examination

After sampling, the colon was fixed in 10% formalin solution. The fixed intestinal tissues were embedded in paraffin, sectioned, stained with hematoxylin and eosin, and graded using the following scoring criteria. Table 2 Histopathological scoring criteria of enteritis inflammatory cell infiltration 0 occasional or no infiltration 1 lamina propria 2 infiltration into the submucosa 3 transmural infiltration Table 3 Histopathological scoring criteria of enteritis tissue damage 0 no mucosal damage 1 focal lesion 2 mucosal erosion or ulceration 3 Extensive damage affecting the submucosa 3. Test results a. Disease activity index (DAI) (Table 3, Fig. 1) Table 3 The impact of continuous administration of test substance compound I on the disease activity index (DAI) score of mice group DAI total score AUC Weight Change AUC Bleeding AUC Stool AUC G1 (normal control) Mean 1.63 1.63 0.00 0.00 Standard Error (SEM) 0.80 0.80 0.00 0.00 G2 (model control) Mean 30.67 8.17 10.21 12.29 Standard Error (SEM) 2.47 0.87 0.88 0.92 G3 (Compound I) Mean 25.67 8.67 7.42 9.58 Standard Error (SEM) 1.26 0.75 0.48 0.62 G4 (Cyclosporine A) Mean 21.63 4.67 6.29 10.67 Standard Error (SEM) 1.74 1.08 0.54 0.43 G5 (tofacitinib) Mean 29.67 9.33 9.29 11.04 Standard Error (SEM) 2.00 1.06 0.73 0.68

The DAI total score is the sum of the three parameter scores of blood in the stool, diarrhea and weight loss. Most of the animals in the model control group had clinical symptoms such as blood in the stool and watery diarrhea, which indicated that the enteritis model in this experiment was successfully established. Compared with the model control group, the positive control cyclosporine A can significantly improve the clinical symptoms, and the area under the curve (AUC) of the total score of the disease activity index DAI was significantly reduced. The test compound I can also significantly reduce the AUC of the total DAI score at a dose of 6 mg/kg/d. The positive control cyclosporine A significantly improved the symptoms of hematochezia in animals, and the test compound Ⅰ also significantly relieved hematochezia at a dose of 6 mg/kg/d, while the reference drug tofacitinib did not significantly improve hematochezia. The AUC analysis of the area under the curve of the DAI stool score showed that the test compound Ⅰ significantly relieved the diarrhea symptoms at 6 mg/kg/d, while the positive controls cyclosporine A and tofacitinib had no significant improvement on the diarrhea symptoms. Therefore, compound I is better than tofacitinib in improving enteritis DAI. B. Pathological evaluation (Table 4, Table 5, Fig. 2) Table 4 Effect of continuous administration of test substance Compound I on pathological score (inflammation) of mice group inflamed G1 (normal control) Mean 0.00 Standard Error (SEM) 0.00 G2 (model control) Mean 2.42 Standard Error (SEM) 0.15 G3 (Compound I, 6mg/kg/d) Mean 2.08 Standard Error (SEM) 0.08 G4 (Cyclosporine A) Mean 2.08 Standard Error (SEM) 0.08 G5 (tofacitinib) Mean 1.92 Standard Error (SEM) 0.15 G6 (Compound I, 20mg/kg/d) Mean 2.00 Standard Error (SEM) 0.00 G7 (Compound I, 60mg/kg/d) Mean 1.83 Standard Error (SEM) 0.11 Table 5 The impact of continuous administration of test substance compound I on the pathological score (injury) of mice group damage G1 (normal control) Mean 0.00 Standard Error (SEM) 0.00 G2 (model control) Mean 2.00 Standard Error (SEM) 0.12 G3 (Compound I) Mean 1.25 Standard Error (SEM) 0.22 G4 (Cyclosporine A) Mean 1.33 Standard Error (SEM) 0.31 G5 (tofacitinib) Mean 1.58 Standard Error (SEM) 0.15

In this study, samples were taken at the end of the trial, and the pathological analysis of the colon was performed to evaluate inflammatory cell infiltration and tissue damage. The pathological results showed that the animals in the model control group had obvious pathological changes, with inflammatory cells infiltrating into the submucosa and even transmural infiltration, and intestinal mucosal erosion or ulceration. Compared with the model control group, the positive control cyclosporine A significantly improved inflammatory cell infiltration and tissue damage. Representative images are shown in Figure 3. The continuous administration of the test compound Ⅰ for 10 days significantly improved the inflammatory response of the colon, and the dose group 6mg/kg/d, 20mg/kg/d and 60mg/kg/d significantly reduced the inflammatory infiltration, and the dose group 6mg/kg/d had a significant effect on colon inflammation. Tissue damage was also significantly improved. Tofacitinib is only effective against inflammatory infiltrates. Therefore, the test compound I is better than tofacitinib in improving the histopathology of enteritis.

In conclusion, compound I significantly improved the disease activity index and histopathology of DAI in the mouse enteritis model, and was superior to tofacitinib.

Figure 1 is the effect of continuous administration of the test compound I on the disease activity index (DAI) of mice. Among them, A is the total score of the disease activity index, B is the weight change score, C is the bleeding score, and D is the stool score; Figure 2 is the effect of continuous administration of the test substance Compound I on the pathological scores of mice. Among them, A is inflammatory cell infiltration, B is tissue damage; Figure 3 is a representative pathological picture of the test compound I. Among them, A is the representative picture of G1 (normal control) (hematoxylin-eosin staining), inflammation: 0, occasional or no infiltration; injury: 0, no mucosal damage; B is the representative picture of G2 (model control) (Hematoxylin-eosin staining), inflammation: 3, transmural infiltration; injury: 2, mucosal erosion or ulceration; C is a representative picture of G3 (compound I) (hematoxylin-eosin staining), inflammation: 2. Infiltration into the submucosa; damage: 1, focal lesion; D is a representative image of G4 (cyclosporine A) (hematoxylin-eosin staining), inflammation: 2, infiltration into the submucosa; damage: 1 , focal lesion; E is a representative image of G5 (tofacitinib) (hematoxylin-eosin staining), inflammation: 2, infiltrating into the submucosa; injury: 1, focal lesion; #, P<0.05; ##, P<0.01; ###, P<0.001; compared between normal control group and model control group; *, P<0.05; **, P<0.01; ***, P<0.001; compared with model control group.

Claims (10)

Use of a compound represented by formula I or a pharmaceutically acceptable salt thereof in the preparation of medicines for treating and/or preventing inflammatory bowel disease;

. The use according to claim 1, characterized in that the inflammatory bowel disease is ulcerative colitis. The use according to claim 1, characterized in that the inflammatory bowel disease is Crohn's disease. The use according to at least one of claims 1-3, characterized in that the inflammatory bowel disease has symptoms of blood in the stool and/or diarrhea. The use according to at least one of claims 1-4, characterized in that the medicine for treating and/or preventing inflammatory bowel disease contains a pharmaceutical adjuvant. The use according to at least one of claims 1-5, characterized in that the compound represented by formula I or a pharmaceutically acceptable salt thereof is one or the only active ingredient of the drug. The use according to at least one of claims 1-6, characterized in that a therapeutically effective amount of the compound represented by formula I or a pharmaceutically acceptable salt thereof is administered to a subject in need. The use as described in claim 7, characterized in that, the administration method is oral administration; And/or, the therapeutically effective amount of administration is 4-100 mg/kg/d, preferably 6-60 mg/kg/d, such as 6 mg/kg/d, 20 mg/kg/d or 60 mg/kg/d ; And/or, the administration frequency is 1-3 times/day, such as 1-2 times/day. A pharmaceutical composition for treating and/or preventing inflammatory bowel disease in a subject, characterized by comprising: a compound represented by formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutical adjuvant;

. The pharmaceutical composition for treating and/or preventing inflammatory bowel disease in a subject according to claim 9, wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease; And/or, the compound represented by formula I or a pharmaceutically acceptable salt thereof is in a therapeutically effective amount.

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