Use of tyrosine kinase inhibitors

Abstract

The present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating mastocytoma in a non-human mammal, and a method for treating mastocytoma in a non-human mammal. [Formula 1] JPEG2026505329000011.jpg46170

Description

[Technical Field]

[0001] This invention claims priority to a Chinese patent application filed with the Intellectual Property Office of the People's Republic of China on February 8, 2023, bearing application number 202310100761.3 and entitled "Use of Tyrosine Kinase Inhibitors," the entire contents of which are incorporated herein by reference.

[0002] The present invention belongs to the field of pharmaceutical chemistry industry, and specifically relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the treatment of mast cell tumors. [Background technology]

[0003] As society develops, more and more people are keeping pets. According to survey data from the 2021 Pet Industry White Paper, there are 58.06 million pet cats and 54.29 million pet dogs in urban households in China. The number of elderly pets and geriatric diseases in pets is also increasing over time, with the incidence of pet tumors in particular becoming significantly higher. Among these, malignant tumors pose a direct threat to the lives and health of pets.

[0004] Animal skin tumors are among the most common tumors. Studies have shown that the incidence of canine skin tumors is approximately one-third of that of canine tumors, with malignant tumors accounting for 20% to 40% of all skin tumors. Skin tumors have a significant impact on the health of pet dogs. Among these, mast cell tumors (MCT) are the most common skin tumor in dogs and the second most common in cats. Mast cell tumors occur in dogs of all ages but are more common in older dogs. This disease does not have a gender predilection. While mixed-breed dogs are more likely to develop this disease, some breeds, such as boxers, Labrador retrievers, pugs, and beagles, are susceptible. Mast cell tumors occur primarily in the mesenchymal tissue of canine skin and present as red, hairless nodules that can occur anywhere on the body.

[0005] Neoplastic proliferation of mast cells is called mastocytoma (MCT), and mast cell tumors are characterized by abnormal proliferation and accumulation of mast cells. Neoplastic mast cell masses spontaneously degranulate, releasing bioactive molecules that can have serious and potentially fatal effects, such as allergic reactions, gastric and duodenal ulcers or perforations, glomerular disease, and bleeding. In dogs, mastocytoma (MCT) exhibits highly variable clinical symptoms and biological behavior. In other species, such as cats, the biological behavior of MCT is relatively less variable. In humans, mastocytoma (MCT) is called mastocytosis and usually manifests as a systemic disease rather than a distinct tumor. Human mastocytosis can range from spontaneously resolving skin lesions to highly invasive, multisystemic malignancies.

[0006] The incidence of canine mast cell tumors is 16% to 21% of all skin tumors, with a malignant rate of 19% to 39%. The metastatic rate of poorly differentiated mast cell tumors is 55% to 96%. They typically metastasize first to the lymph nodes, then to the spleen and liver, and finally to the bone marrow. At this stage, large numbers of mast cells are found in the peripheral blood, a condition known as mastocythemia. Dogs with malignant mast cell tumors have poor survival and quality of life. With the gradual application of molecular research methods to small animal clinical practice, research has revealed that approximately 15% to 40% of canine mast cell tumors contain c-kit gene mutations. Currently, identified mutations include point mutations, internal tandem duplications (ITDs), and base deletion mutations. A large body of literature has demonstrated that c-kit mutations are significantly associated with the development and progression of canine mast cell tumors. Mast cell tumors include cutaneous and systemic mast cell tumors. Systemic mast cell tumors are diseases that affect multiple organs or systems due to the neoplastic proliferation of mast cells, and may or may not include cutaneous mast cell tumor pathology. Examples include visceral and intestinal mast cell tumors.

[0007] The Patnaik grading system is commonly used to pathologically grade canine mast cell tumors. In the Patnaik grading system, mast cells in grade I MCT are well differentiated, neatly dispersed among the collagen fibers of the dermis, have ample cytoplasmic space and filled with granules, condensed nuclear chromatin, and no obvious nucleoli or mitotic cells. Grade II MCT mast cells are well differentiated, have ample cytoplasmic space and filled with granules, have mild cytoplasmic atypia, some porous nucleoli are evident, some nucleoli are bleb-like, and an appropriate number of mitotic cells are present. Grade III MCT mast cells have hazy cytoplasm, moderate to severe atypia, deeply stained nuclei, obvious nucleoli, and mitotic cells are present.

[0008] While canine MCTs often originate in the skin or subcutaneous tissue, feline MCTs differ from dogs in that they fall into three typical categories, each with some of the same pathology: cutaneous, splenic / visceral, and intestinal. Although the etiology is still unclear, feline MCTs also have clear c-kit activating mutations.

[0009] Regarding treatment options, a combination of surgery and radiation therapy is common for dogs with localized, solitary mast cell tumors without distant metastasis, but this treatment method is not widely accepted by many veterinary clinics and pet owners due to the high cost of radiation therapy equipment and treatment. On the other hand, for dogs with distant metastasis or highly aggressive mast cell tumors, chemotherapy is required.

[0010] Tyrosine kinases are a popular target in research on human antitumor drugs. Because abnormalities in this target pathway can also lead to tumor development in pets, tyrosine kinase inhibitors could theoretically be used to treat pet tumors. Currently, tyrosine kinase inhibitors used in pet tumor treatment, including toceranib and masitinib, show particularly pronounced efficacy in MCTs with c-kit gene mutations. The mechanism of action of these drugs is to competitively inhibit the binding of ATP to mutated tyrosine kinase receptors, thereby inhibiting phosphorylation of the tyrosine kinase receptor's activation center, thereby inhibiting its activation and ultimately inhibiting tumor cell proliferation.

[0011] WO2011153814A1 discloses that N-(5-((Z)-(5-fluoro-2-carbonylindol-3-ylidene)methyl)-2,4-dimethyl-1H-pyrrol-3-yl)-3-(4-methylpiperazin-1-yl)propionamide compounds have the potential to be developed as antitumor drugs, but their therapeutic efficacy against tumors in non-human mammals (e.g., pets) has not yet been demonstrated. Summary of the Invention

[0012] The present invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating mast cell tumors in a non-human mammal. The specific contents are as follows:

[0013] The present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating mast cell tumors in a non-human mammal, comprising: [ka] The compound represented by formula (I) is N-(5-((Z)-(5-fluoro-2-carbonylindol-3-ylidene)methyl)-2,4-dimethyl-1H-pyrrol-3-yl)-3-(4-methylpiperazin-1-yl)propionamide.

[0014] In some preferred embodiments of the present invention, the mastocytoma referred to in the above uses includes cutaneous mastocytoma and systemic mastocytoma.

[0015] In some preferred forms of the invention, the mastocytoma according to the above uses is selected from cutaneous mastocytoma.

[0016] In some preferred forms of the invention, the mastocytoma according to the above uses is selected from recurrent mastocytoma.

[0017] In some preferred forms of the invention, the mastocytoma according to the above uses is selected from unresectable, recurrent grade 2 or grade 3 cutaneous mastocytoma.

[0018] In some preferred embodiments of the present invention, the mast cell tumors described in the above uses are tumors caused by mutations in the c-kit gene.

[0019] In some preferred embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above uses is administered to a non-human mammal at a dose of 0.1 to 10 mg / kg at least once a day or once every other day.

[0020] In some preferred embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above uses is administered to a non-human mammal at a dose of 1 to 5 mg / kg at least once daily or once every other day.

[0021] In some preferred embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above uses is administered to a non-human mammal at a dose of 1.5 to 3.5 mg / kg at least once daily or once every other day.

[0022] In some preferred embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above uses is administered to a non-human mammal at a dose of 2 to 3.5 mg / kg at least once daily or once every other day.

[0023] In some preferred forms of the invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above uses is administered to a non-human mammal at least once a day or once every other day in one or more of the following dosages: 2.1 mg / kg, 2.2 mg / kg, 2.3 mg / kg, 2.4 mg / kg, 2.5 mg / kg, 2.6 mg / kg, 2.7 mg / kg, 2.8 mg / kg, 2.9 mg / kg, 3.0 mg / kg, 3.1 mg / kg, 3.2 mg / kg, 3.3 mg / kg, 3.4 mg / kg, and 3.5 mg / kg.

[0024] In some preferred embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above uses is administered to a non-human mammal by any one of the following routes of administration: oral, parenteral, rectal, transdermal, intranasal or inhalation routes, and intratumoral administration.

[0025] In some preferred forms of the invention, the pharmaceutically acceptable salts of the compounds of formula (I) described in the above uses are selected from dimaleate salts.

[0026] In some preferred forms of the invention, the non-human mammals described in the above uses are selected from dogs, cats, rats and horses.

[0027] Another aspect of the present invention further provides a method for treating mastocytoma in a non-human mammal, comprising administering to said non-human mammal a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. [ka]

[0028] In some preferred embodiments of the present invention, the mastocytoma described in the above treatment methods includes cutaneous mastocytoma and systemic mastocytoma.

[0029] In some preferred forms of the invention, the mastocytoma described in the above methods of treatment is selected from cutaneous mastocytoma.

[0030] In some preferred forms of the invention, the mastocytoma described in the above methods of treatment is selected from recurrent mastocytoma.

[0031] In some preferred forms of the invention, the mastocytoma described in the above methods of treatment is selected from unresectable, recurrent grade 2 or grade 3 cutaneous mastocytoma.

[0032] In some preferred embodiments of the present invention, the mastocytoma described in the above treatment methods is a tumor caused by a mutation in the c-kit gene.

[0033] In some preferred embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above method of treatment is administered to a non-human mammal at a dose of 0.1 to 10 mg / kg at least once a day or once every other day.

[0034] In some preferred embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above method of treatment is administered to a non-human mammal at a dose of 1 to 5 mg / kg at least once daily or once every other day.

[0035] In some preferred embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above-mentioned method of treatment is administered to a non-human mammal at a dose of 1.5 to 3.5 mg / kg at least once a day or once every other day. In some preferred embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above-mentioned method of treatment is administered to a non-human mammal at a dose of 2 to 3.5 mg / kg at least once a day or once every other day.

[0036] In some preferred embodiments of the present invention, the compound of Formula (I) or a pharmaceutically acceptable salt thereof described in the above methods of treatment is administered to a non-human mammal at least once a day or once every other day at one or more of the following dosages: 2.1 mg / kg, 2.2 mg / kg, 2.3 mg / kg, 2.4 mg / kg, 2.5 mg / kg, 2.6 mg / kg, 2.7 mg / kg, 2.8 mg / kg, 2.9 mg / kg, 3.0 mg / kg, 3.1 mg / kg, 3.2 mg / kg, 3.3 mg / kg, 3.4 mg / kg, and 3.5 mg / kg.

[0037] In some preferred embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the above method of treatment is administered to a non-human mammal by any one of the following routes of administration: oral, parenteral, rectal, transdermal, nasal or inhalation routes, and intratumoral administration.

[0038] In some preferred forms of the invention, the pharmaceutically acceptable salts of the compounds of formula (I) described in the above methods of treatment are selected from dimaleate salts.

[0039] In some preferred forms of the invention, the non-human mammal described in the above methods of treatment is selected from dogs, cats, rats and horses.

[0040] Another aspect of the present invention further provides a compound of formula (I) below, or a pharmaceutically acceptable salt thereof, for use in the treatment of mast cell tumors in non-human mammals. [ka]

[0041] Definitions and Explanations Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings: A particular term or phrase, unless specifically defined, should not be understood as being indefinite or uncertain, but rather should be understood in its ordinary sense.

[0042] The term "pharmaceutically acceptable" means that those compounds, materials, compositions and / or dosage forms are, within the scope of sound medical judgment, suitable for use in contact with animal tissues without excessive toxicity, irritation, allergic response, or other problem or complication, and are consistent with a reasonable benefit / risk ratio.

[0043] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention prepared from a compound discovered in this invention having a specific substituent and a relatively non-toxic acid or base. When a compound of the present invention has a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of base in pure solution or in a suitable inert solvent. When a compound of the present invention has a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of acid in pure solution or in a suitable inert solvent. Pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds having acidic or basic groups by conventional chemical methods. Such salts are generally prepared by reacting the free acid or free base form of such compounds with the stoichiometrically appropriate base or acid in water, an organic solvent, or a mixture of both. Specifically, pharmaceutically acceptable salts are described in detail in, for example, S.M. Berge, et al., J. Pharmaceutical Sciences, 1977, 66:1. Representative acid addition salts include, but are not limited to, acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, sulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, caproate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate (isothiosulfate), lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, palmitate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, phosphate, malate, glutamate, bicarbonate, p-toluenesulfonate, and undecanoate salts.Similarly, basic nitrogen-containing groups can be quaternized with materials such as lower alkyl halides, such as chlorides, bromides, and iodides of methyl, ethyl, propyl, and butyl groups; dialkyl sulfates, such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate, and dipentyl sulfate; long chain halides, such as chlorides, bromides, and iodides of decyl, dodecyl, tetradecyl, and octadecyl groups; aryl alkyl halides, such as benzyl bromide, phenylethyl bromide, and the like.

[0044] The term "therapeutically effective amount" refers to a sufficient amount of a compound of the present invention or a pharmaceutically acceptable salt thereof for any medical treatment and / or prophylaxis, suitable for treating a disorder at a reasonable benefit / risk ratio. However, it should be recognized that the total daily usage of the pharmaceutically acceptable salts of the compound of formula (I) and compositions of the present invention should be determined by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend on a variety of factors. Such factors include the disorder to be treated and its severity; the activity of the specific compound used; the specific composition used; age, weight, general health, sex, and diet; the time, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or simultaneously with the specific compound used; and similar factors well known in the medical field.

[0045] The term "administration" refers to the physical introduction of a composition containing a therapeutic agent into a subject using any of a number of methods and delivery systems known to those skilled in the art. Routes of administration include oral, parenteral, intraperitoneal, intravenous, intraarterial, transdermal, sublingual, intramuscular, rectal, buccal, intranasal, inhalation, vaginal, intraocular, topical administration (e.g., via a catheter or stent), subcutaneous, intraadipose, intraarticular, intraperitoneal, and intrathecal. In some embodiments of the invention, "administration" refers to delivery to a subject via the oral route.

[0046] As used herein, the term "non-human mammal" refers to a mammal other than a human at any stage of development. In some embodiments, the non-human mammal includes, but is not limited to, a rodent (e.g., a mouse, a rat, a rabbit), a dog, a cat, a sheep, a cow, a horse, a primate (e.g., a monkey), and / or a pig.

[0047] As used herein, "recurrent grade 2 or grade 3 cutaneous mast cell tumor" refers to recurrent Patnaik grade II or grade III cutaneous mast cell tumor.

[0048] Tumor recurrence generally refers to the phenomenon in which a similar tumor appears at the primary site or its surrounding area, or at another site in the body, after a confirmed tumor has been cured by radical treatment (including surgery, radiation therapy, chemotherapy, and other treatments). Tumor recurrence includes local recurrence and distant metastasis.

[0049] Beneficial effects of the present invention: The compound of formula (I) or its pharmaceutically acceptable salt of the present invention can significantly inhibit the growth of mast cell tumors in non-human mammals, prolong the survival time of the animals, and reduce the types and incidence of side effects. The compound of formula (I) or its pharmaceutically acceptable salt of the present invention has relatively good efficacy and safety in treating mast cell tumors in non-human mammals. [Brief explanation of the drawings]

[0050] The drawings described herein are intended to provide a further understanding of the present invention and constitute a part of the present invention. The illustrative embodiments of the present invention and the description thereof are intended to interpret the present invention and are not to be construed as an undue limitation of the present invention. [Figure 1] Figure 1 shows a photograph (2.5 x 2.2 cm) of the baseline tumor measurement on October 3rd in the golden retriever case in Experimental Example 3. [Figure 2]FIG. 2 is a photograph (2.2×2.1 cm) of the therapeutic effect of the compound represented by formula (I) on the golden retriever case in Experimental Example 3 on October 17th. [Figure 3] FIG. 3 is a photograph showing the therapeutic effect of the compound represented by formula (I) on November 1st in the golden retriever case in Experimental Example 3 (the white circle indicates the tumor). DETAILED DESCRIPTION OF THE INVENTION

[0051] The present invention will be described in detail below through examples, but these examples are for illustrative purposes only and do not limit the scope of the present invention. Likewise, the present invention is not limited to any specific preferred embodiments described herein. Those skilled in the art should understand that equivalent substitutions and corresponding improvements made to the technical features of the present invention still fall within the scope of protection of the present invention. Unless otherwise specified, all examples are carried out according to normal experimental conditions.

[0052] The compound of formula (I) of the present invention or its salt can be prepared by referring to the methods described in patents WO2011153814A1, CN104119321A, or other known methods in the art.

[0053] Experimental Example 1 Growth inhibitory effect of the compound of formula (I) of the present invention on tumor cells Test sample: dimaleate of the compound of formula (I) of the present invention. The inhibitory effect of the dimaleate salt of the compound represented by formula (I) on the growth of mouse mastocytoma cells P815 (ATCC-TIB-64) was detected by MTT method (thiazolyl blue colorimetric method), and the IC 50 (50% inhibitory concentration) was calculated.

[0054] [Table 1A]

[0055] Experimental Example 2 Evaluation of in vivo efficacy of the compound of formula (I) of the present invention on DBA / 2 mouse model of subcutaneous syngeneic transplant tumor of mouse mastocytoma P815 cells Test samples: dimaleate of the compound of formula (I) of the present invention (content: 99.7%), toceranib phosphate (content: 98.2%, commercially available). The concentrations and dosages of the test samples prepared in the following examples were all calculated based on the free base. Test animals: The species were mice, the strain was DBA / 2 mice, the age was 6-8 weeks, the weight was 19-24 grams, and the sex was female.

[0056] Experimental Methods and Steps: Mouse mastocytoma P815 cells (ATCC-TIB-64) were cultured in vitro as adherent cells in DMEM medium, 10% fetal bovine serum, and 1% bispecific antibody in a 37°C, 5% CO2 cell culture incubator. Cells were passaged twice a week using trypsin-EDTA digestion. When the cell confluency reached 80%-90% and the required number of cells was met, the cells were harvested, counted, and seeded.

[0057] Each mouse received 5 × 10 subcutaneous injections into the axilla of the right forelimb. 5 P815 cells were subcutaneously inoculated in 0.1 mL of PBS. Eight days after cell inoculation, the tumor volume reached an average of 100 mm. 3 Once the serotonin concentration reached 100 mg / kg, the mice were divided into groups and treatment began. The experiment consisted of a negative control group (containing no active ingredient), a treatment group, and a positive control group (toceranib as the active ingredient), each consisting of eight mice. The negative control (solvent), the dimaleate salt of compound of formula (I), and toceranib phosphate were orally administered once daily for eight days. The specific preparation method is shown in Table 1, and the dosing schedule is shown in Table 2. The animals were monitored daily for health and mortality. Regular inspections included observing daily behavior, such as activity, food and water intake (visual observation only), changes in body weight (measured twice a week), and the effects of tumor growth and drug treatment on external appearance, or any other abnormalities.

[0058] [Table 1]

[0059] [Table 2]

[0060] The experimental endpoint was to consider whether tumor growth was inhibited, delayed, or cured. The diameter of the tumor was measured with a caliper. The formula for calculating tumor volume was: V = 0.5a × b 2 where a and b represent the long and short diameters of the tumor, respectively. The tumor inhibitory effect of a compound is expressed as TGI (%). TGI (%) represents the tumor growth inhibition rate. The calculation formula for TGI (%) is TGI (%) = [1 - (mean tumor volume at the end of administration for a treatment group - mean tumor volume at the start of administration for that treatment group) / (mean tumor volume at the end of treatment for the solvent control group - mean tumor volume at the start of treatment for the solvent control group)] × 100%. The results of the experiment are shown in Table 3.

[0061] [Table 3] On the 8th day after administration, the mean tumor volume of the tumor-bearing mice in the vehicle control group was 825 mm 3 Compared with the vehicle control group, the average tumor volume was 483 mm when the test substance, the compound represented by formula (I), was administered at a dose of 20 mg / kg and when toceranib was administered at a dose of 40 mg / kg. 3 (TGI=47.25%, p=0.003) and 510mm 3 (TGI=43.38%, p<0.001).

[0062] As can be seen from the experimental results, even when the compound represented by formula (I) of the present invention is administered at half the dose of toceranib, it can achieve the same therapeutic effect as toceranib, reduce the therapeutically effective dose and administration cost, and is advantageous in reducing the side effects of the drug, improving its safety.

[0063] Experimental Example 3 Evaluation of the antitumor effect and side effects of the compound represented by formula (I) of the present invention on canine mast cell tumors Research method: From May 2021, a total of 14 dogs with mastocytoma were collected, grouped, and treated with the dimaleate salt of the compound of formula (I).

[0064] [Table 4] Canine patients with mast cell tumors were orally administered 2.0 mg / kg to 3.25 mg / kg of the dimaleate salt of the compound of formula (I) (all doses calculated based on the free base; the same applies below) once every other day. A baseline assessment was performed on canine patients with evaluable solid tumors before the first administration of the study drug. After the first administration of the study drug, tumor measurements and general blood tests were performed every two weeks to evaluate tumor response to treatment, and blood biochemistry and imaging tests were performed every four weeks until progression, discontinuation due to intolerance to treatment-related side effects, or death of the dog in the group. The therapeutic efficacy and safety of treatment with the compound of formula (I) for canine mast cell tumors and mammary tumors were evaluated. The main observational endpoints were overall response rate (ORR), disease control rate (DCR), progression-free survival (PFS) and overall survival (OS); complete response (CR), partial response (PR), stable disease (SD) and progressive disease (PD).

[0065] (1) A case of mast cell tumor in a golden retriever The golden retriever presented on September 26, 2021, with a tumor in the hock of its right hind leg. During physical examination, palpation revealed that the popliteal lymph nodes in the right hind leg were larger than those on the left, and tumor cytology revealed mast cell tumors. On October 3, blood routine tests, blood biochemistry tests, abdominal ultrasound, and liver and spleen FNA were performed. No liver or spleen metastases were detected, and the clinical stage was stage II. Sampling from the tumor site revealed no mutations in the 11th exon of the c-kit gene. The baseline measurement of the tumor in the right hind leg was 2.5 × 2.2 cm (shown in Figure 1). The patient dog began receiving the dimaleate salt of the compound of formula (I) on October 3rd. After 14 days of treatment at an initial dose of 3.07 mg / kg, the dog was re-examined (October 17th). The tumor measured 2.2 x 2.1 cm (shown in Figure 2), a 12% decrease compared to baseline, and the tumor response was evaluated as unchanged (SD).

[0066] The patient was reexamined on November 1st, and the tumor on the right hind leg measured 2 x 1 cm (see Figure 3). The tumor thickness had decreased significantly, and the tumor response was assessed as partial response (PR). Ultrasound examination of the popliteal lymph nodes revealed that the right popliteal lymph node (0.78 x 0.52 cm) was slightly larger than the left (0.68 x 0.29 cm). Because the tumor had decreased significantly, the owner considered surgical removal. The patient underwent resection of the tumor in the hock of the right hind leg and removal of the popliteal lymph nodes in the right hind leg on November 3rd. Pathological examination confirmed a definitive diagnosis of mast cell tumor with popliteal lymph node metastasis, and the disease was staged as Grade II (a three-grade classification method). The postoperative tumor margin was assessed as a margin of deeply infiltrated tumor cells, and the patient began treatment with the dimaleate salt of compound of formula (I) at a dose of 2.63 mg / kg on December 5th.

[0067] The patient dog was re-examined on days 47, 82, 117, and 145 after surgery, and no recurrence of the tumor was found on the skin. As of March 28, 2022, the patient dog had received a total of 66 doses of the drug, was in good mental condition, had a good appetite, and survived for over 176 days.

[0068] (2) A case of mast cell tumor in a Tibetan mastiff The patient presented with a tumor in the right hind leg and was diagnosed cytologically as mast cell tumor, clinically staged as stage II, without c-kit mutation.

[0069] On May 24, 2021, the patient was treated with the dimaleate salt of compound of Formula (I) at a dose of 2.84 mg / kg. The baseline tumor size was 7.1 x 4 x 4.5 cm. On June 21, the patient was reexamined, revealing a tumor size of 6.6 x 3.9 x 4.4 cm. Repeated blood tests and serum biochemistry tests revealed no significant abnormalities compared to baseline, and treatment was continued at the 2.84 mg / kg dose. On July 19, the patient was reexamined, revealing a tumor size of 5.5 x 3.5 x 4.5 cm. Treatment was continued at the 2.84 mg / kg dose. The patient died on September 18, 2021, with a total survival time of 117 days.

[0070] (3) A case of mast cell tumor in a Labrador retriever One year prior, the patient presented with a tumor in his right armpit. Cytological findings indicated a mast cell tumor, and ultrasound revealed a large tumor at the tip of the spleen. Resection of the tumor and lymph nodes in his right armpit, as well as splenectomy, were performed. Pathologically, the splenic tumor was diagnosed as angiosarcoma, and the tumor in his armpit was diagnosed as a mast cell tumor. The pathological classification of the mast cell tumor was grade III (a classification system with three grades). Five cycles of doxorubicin chemotherapy were administered one month after surgery.

[0071] The patient returned to the clinic seven months after surgery (December 13, 2021) due to the development of new tumors on her back and vaginal orifice. Cytological examination revealed that both tumors were mast cell tumors. No other obvious metastatic lesions were found, and the clinical stage was classified as stage III. Sampling of the tumor sites on the back and vaginal orifice revealed a 45-bp ITD mutation at the end of exon 11 of the c-kit gene. The back tumor was designated the target lesion, and the baseline measurement was 1.5 cm.

[0072] The patient dog was treated with the dimaleate salt of the compound of formula (I) at an initial dose of 2.42 mg / kg for 14 days from December 13th. On December 26th, the dog returned to the hospital for a reexamination. The tumor on the back had disappeared and was no longer measurable, and the tumor response was assessed as CR. The tumor on the vaginal orifice side had clearly decreased.

[0073] The patient dogs were reexamined on days 42 and 77 of treatment, and no recurrence of the dorsal tumor was observed in either case, resulting in a CR. No growth of the tumor on the vaginal orifice was observed.

[0074] The patient returned for reexamination on day 126 of treatment. No dorsal tumor recurrence was noted, and the tumor proximal to the vagina had become red and enlarged, measuring 1.74 × 1.9 cm. Target lesion tumor response was assessed as CR, and non-target lesion response was assessed as PD.

[0075] The patient is still undergoing treatment, and no recurrence of the tumor on the back has been observed, nor has any new lesion been observed. The overall therapeutic effect of the target lesion was assessed as CR. As of April 17, 2022, the patient dog had received a total of 51 doses of the drug. The patient's mental state and appetite were good, and the dog's survival period exceeded 126 days.

[0076] (4) A case of mast cell tumor in a golden retriever The patient presented to our hospital on January 2, 2022, with a nasal skin tumor. Histological findings revealed a mast cell tumor, clinical stage I, and no c-kit mutations. Treatment was administered with the dimaleate salt of compound (I) at a dose of 2.4 mg / kg, with no apparent adverse events. The patient was reexamined on February 13. Repeated blood and serum biochemistry tests revealed no obvious abnormalities. Treatment was continued at a dose of 2.4 mg / kg, with symptoms of loose stools and diarrhea appearing after administration. The patient was reexamined on April 10. Repeated blood and serum biochemistry tests revealed no obvious abnormalities. Treatment was continued at a dose of 2.3 mg / kg, with no apparent adverse events after administration. The patient was reexamined on June 6. Nasal ultrasound revealed possible invasion of the nasal cavity by a subcutaneous tumor. Repeated blood and serum biochemistry tests revealed no obvious abnormalities. Treatment was continued at a dose of 2.3 mg / kg, with no apparent side effects observed after administration. The patient was re-examined on August 7th. Nasal ultrasound revealed a subcutaneous tumor with invasive changes to the adjacent nasal turbinates. Repeated blood tests revealed no obvious abnormalities. Treatment was continued at a dose of 2.4 mg / kg. The patient is still being treated with the dimaleate drug of the compound of formula (I), and has survived for over 259 days.

[0077] (5) A case of mast cell tumor in a golden retriever The patient had a tumor on the skin of the left hind leg that was cytologically diagnosed as mast cell tumor. The baseline tumor size was 3.5 x 3.0 cm and the clinical stage was classified as stage III. On September 3, 2022, the patient was treated with the dimaleate salt of compound of formula (I) at a dose of 3.25 mg / kg, with one episode of loose stools following administration. On September 7, the dose was adjusted to 2.75 mg / kg, and treatment continued without any significant side effects. On October 15, the tumor size measured 3.0 x 2.9 cm. During this period, loose and watery stools appeared. The dose was adjusted to 2.25 mg / kg, and treatment continued without any significant side effects following administration. On November 6, the tumor size measured 2.3 x 2.5 cm. The patient is still undergoing treatment. As can be seen from the results of the above animal clinical trials, the compound of formula (I) of the present invention and its salts can effectively inhibit the growth of mast cell tumors and prolong the survival time of animals.

[0078] (6) Occurrence of side effects All treatment-related clinical side effects in this study were grade 1 or 2 and included diarrhea (n=9, 64.3%), bloody stool (n=4, 28.6%), loss of appetite (n=4, 28.6%), vomiting (n=3, 21.4%), and occasional hematuria (n=1, 7.1%). No grade 3 or 4 side effects were observed. (The criteria for evaluating adverse events are based on the Veterinary Cooperative Oncology Group-Common Terminology Criteria for Adverse Events (VCOG-CTCAE) following chemotherapy or biological antineoplastic therapy in dogs and cats v1.1. Vet Comp Oncol. 2016.)

[0079] However, according to the summary of side effects provided in the Palladia® instructions for toceranib phosphate tablets, the total incidence of diarrhea of ​​all grades was 46%, of which Grade 3 or Grade 4 side effects accounted for 6.9%; the total incidence of anorexia of all grades was 39.1%, of which Grade 3 or Grade 4 side effects accounted for 6.9%; the total incidence of vomiting of all grades was 32.2%, of which Grade 3 or Grade 4 side effects accounted for 9.2%; and the total incidence of bloody stool / gastrointestinal bleeding / bloody diarrhea of ​​all grades was 12.6%, of which Grade 3 or Grade 4 side effects accounted for 2.3%.

[0080] As can be seen from the above results, the gastrointestinal side effects of the compound represented by formula (I) and its salts are all grade 1 to 2, whereas toceranib exhibits side effects of grade 3 to 4. It has been revealed that the compound represented by formula (I) and its salts have a lower level of side effects and are more safe.

[0081] The above are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made without departing from the spirit and principle of the present invention are all within the scope of protection of the present invention.

Claims

1. 1. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating mast cell tumors in a non-human mammal, comprising: 【Chemistry 1】 The compound represented by formula (I) is N-(5-((Z)-(5-fluoro-2-carbonylindol-3-ylidene)methyl)-2,4-dimethyl-1H-pyrrol-3-yl)-3-(4-methylpiperazin-1-yl)propionamide.

2. The use according to claim 1 , wherein the mastocytoma includes cutaneous mastocytoma and systemic mastocytoma.

3. The use according to claim 1 or 2, wherein the mastocytoma is selected from cutaneous mastocytoma.

4. The use according to any one of claims 1 to 3, wherein the mastocytoma is selected from recurrent mastocytoma.

5. The use according to any one of claims 1 to 4, wherein the mastocytoma is selected from unresectable, recurrent grade 2 or grade 3 cutaneous mastocytoma.

6. The use according to any one of claims 1 to 5, wherein the mastocytoma is a tumor caused by a mutation in the c-kit gene.

7. The use according to any one of claims 1 to 6, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered to a non-human mammal at a dose of 0.1 to 10 mg / kg at least once a day or once every other day.

8. The use according to any one of claims 1 to 7, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered to a non-human mammal at a dose of 1 to 5 mg / kg at least once a day or once every other day.

9. The use according to any one of claims 1 to 8, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered to a non-human mammal at a dose of 1.5 to 3.5 mg / kg at least once a day or once every other day.

10. The use according to any one of claims 1 to 9, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered to a non-human mammal at a dose of 2 to 3.5 mg / kg at least once a day or once every other day.

11. The use according to any one of claims 1 to 10, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered to a non-human mammal by any one of oral, parenteral, rectal, transdermal, nasal or inhalation routes, and intratumoral administration.

12. The use according to any one of claims 1 to 11, wherein the pharmaceutically acceptable salt of the compound of formula (I) is selected from the dimaleate salt.

13. The use according to any one of claims 1 to 12, wherein the non-human mammal is selected from dogs, cats, rats and horses.

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