QUINOLINE DERIVATIVE FOR TREATING RHEUMATOID ARTHRITIS

Abstract

A quinoline derivative for treating rheumatoid arthritis and a use thereof in preparing a pharmaceutical composition for treating rheumatoid arthritis. In particular, a use of the quinoline derivative 1-[[[4-(4-Fluoro-2-methyl-1H-indol-5-yl)oxy-6-methoxyquinoline-7-yl]oxy]methyl]cyclopropanamine in treating rheumatoid arthritis.

Claims

1-10. (canceled)

11. A pharmaceutical composition for treating rheumatoid arthritis, comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, ##STR00002##

12. The pharmaceutical composition according to claim 11, further comprising any other one or more disease modifying antirheumatic drugs.

13. A method for treating rheumatoid arthritis, comprising administering to a patient in need of treatment a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof ##STR00003##

14. The method according to claim 13, wherein the patient has previously received treatment with a disease modifying antirheumatic drug.

15. The method according to claim 13, wherein the patient is insufficiently responsive or intolerant to at least one disease modifying antirheumatic drug.

16. The method according to claim 13, wherein the patient does not meet target or does not reduce disease activity after having previously received treatment with a traditional disease modifying antirheumatic drug, a targeted disease modifying antirheumatic drug and/or a biological disease modifying antirheumatic drug.

17. The pharmaceutical composition according to claim 11, wherein the rheumatoid arthritis is active rheumatoid arthritis.

18. The pharmaceutical composition according to claim 11, wherein the rheumatoid arthritis is moderate-to-severe, moderate and/or severe active rheumatoid arthritis.

19. The pharmaceutical composition according to claim 11, wherein the compound of formula I or the pharmaceutically acceptable salt thereof is administered as the only active ingredient to a patient.

20. The pharmaceutical composition according to claim 11, further comprising other disease modifying antirheumatic drugs, wherein the compound of formula I or the pharmaceutically acceptable salt thereof is administered simultaneously or sequentially with the other disease modifying antirheumatic drugs to a patient with rheumatoid arthritis.

21. The pharmaceutical composition according to claim 20, wherein the other disease modifying antirheumatic drugs are selected from the group consisting of non-biological disease modifying antirheumatic drugs.

22. The method according to claim 13, wherein the compound of formula I or the pharmaceutically acceptable salt thereof is administered as the only active ingredient to a patient.

23. The method according to claim 13, further comprising administering other disease modifying antirheumatic drugs, wherein the compound of formula I or the pharmaceutically acceptable salt thereof is administered simultaneously or sequentially with the other disease modifying antirheumatic drugs to a patient with rheumatoid arthritis.

24. The method according to claim 23, wherein the other disease modifying antirheumatic drugs are selected from the group consisting of non-biological disease modifying antirheumatic drugs.

25. The method according to claim 23, wherein the other disease modifying antirheumatic drugs are selected from the group consisting of one or more of traditional disease modifying antirheumatic drugs, targeted disease modifying antirheumatic drugs and biological disease modifying antirheumatic drugs.

26. The method according to claim 23, wherein the other disease modifying antirheumatic drugs are selected from the group consisting of methotrexate.

27. The method according to claim 23, wherein the other disease modifying antirheumatic drugs are selected from the group consisting of: methotrexate+sulfasalazine; methotrexate+leflunomide; methotrexate+hydroxychloroquine; sulfasalazine+leflunomide; methotrexate+sulfasalazine+hydroxychloroquine; sulfasalazine+leflunomide+hydroxychloroquine; leflunomide+hydroxychloroquine; sulfasalazine+leflunomide+hydroxychloroquine; and tofacitinib and one or more selected from the group consisting of: methotrexate, sulfasalazine, leflunomide and hydroxychloroquine.

28. The method according to claim 13, wherein the pharmaceutically acceptable salt is a salt formed by the compound of formula I and any of the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, trifluoroacetic acid, propionic acid, hexanoic acid, heptanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, p-chlorobenzenesulfonic acid, p-toluenesulfonic acid, 3-phenylpropionic acid, trimethylacetic acid, t-butylacetic acid, dodecyl sulfuric acid, gluconic acid, glutamic acid, hydroxyl naphthoic acid, salicylic acid and stearic acid; preferably, the pharmaceutically acceptable salt is a hydrochloride salt; and more preferably, the pharmaceutically acceptable salt is a dihydrochloride salt.

29. The method according to claim 13, wherein the rheumatoid arthritis is active rheumatoid arthritis.

30. The method according to claim 13, wherein the rheumatoid arthritis is moderate-to-severe, moderate and/or severe active rheumatoid arthritis.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0077] FIG. 1 illustrates the effect of drugs on the weight of the rats;

[0078] FIG. 2 illustrates the effect of the drugs on the width of the hind limb ankle joints of the rats;

[0079] FIG. 3 illustrates the effect of the drugs on the thickness of the hind limb ankle joints of the rats;

[0080] FIG. 4 illustrates the effect of the drugs on the scores of the hind limb joints of the rats;

[0081] FIG. 5 illustrates the effect of the drugs on the weight of the hind limb joints of the rats;

[0082] FIG. 6 illustrates front photographs of swelling of hind limb joints of the animals in each group; and

[0083] FIG. 7 illustrates side view photographs of swelling of hind limb joints of the animals in each group.

DETAILED DESCRIPTION

EXAMPLE 1

[0084] Cell Assay

[0085] An MTT assay for the inhibition of human rheumatoid arthritis synovial cells using the dihydrochloride salt of the compound of formula I as the test drug: the trypsinized cell suspension was seeded in a 96-well plate at a volume of 100 μL per well, with 1×10.sup.4 cells per well. The edges of the plate were wiped with sterile PBS. The plate was placed in a cell incubator and incubated at 37° C. with 5% CO.sub.2. After 24 h of incubation, the original culture solution in the wells was aspirated. Then 100 μL of culture solution with gradient concentration of drug was added to each well, and 6 concentrations were set, namely, 0 μM, 0.5 μM, 1 μM, 2 μM, 5 μM and 10 μM, respectively. The plate was placed in the incubator and incubated at 37° C. with 5% CO.sub.2. After 24 h and 48 h after dosing, 20 μL of MTS was added to each well. The plate was incubated at 37° C. for 1.5 h. The absorbance of each well at a wavelength of 540 nm was measured by a microplate reader, and the inhibition rate of cells in each group was calculated.

[0086] The calculation formula is as follows:

cell inhibition rate/%=(1−OD value of blank control group/OD value of each concentration drug group)×100%

[0087] The results showed that compared with the drug-free group at 0 μM, the drug-containing group could significantly inhibit the proliferation of human rheumatoid arthritis synovial cells, and the inhibitory effect was enhanced along with the increase of the concentration of the drug.

EXAMPLE 2

[0088] Animal Experiments

[0089] 1. Establishment of CIA Model

[0090] After adaptive feeding of 80 SPF male SD rats with the weight of 140±10 g for 5 days, 10 male SD rats were randomly selected as a normal control group, and the rest were subjected to collagen-induced arthritis model modeling. The specific methods are as follows.

[0091] A solution of the bovine type II collagen (CII) in acetic acid (containing 2.0 mg.Math.mL.sup.−1 bovine CII) and an equal volume of complete Freund's adjuvant (CFA) were completely mixed, and the mixture was fully emulsified. The emulsion was added dropwise to water without diffusing as the degree, and finally the emulsion containing 1.0 mg.Math.mL.sup.−1 bovine CII was prepared. 0.2 mL of the emulsion was taken and injected intradermally into the dorsal and the base of the tail at four points (two points on the dorsal and two points on the base of the tail) for the primary immunization (d0). After day 7 (d7) of primary immunization, the emulsion was injected again as described above to boost the immunization. Then the arthritis index (AI) was scored on day 14 (d14). The highest score of each rat was 16 points, and the arthritis induction was considered successful if there was at least 1 paw with AI score≥2.

[0092] 2. Grouping and Administration

[0093] The CIA rats successfully modeled were randomly divided into a drug medium-dose group, a drug low-dose group and a model group, with 10 rats in each group. The normal group and the model group were subjected to intragastric administration with 0.02 mL.Math.g.sup.−1 normal saline; the drug medium-dose group and the drug low-dose group were administered with the dihydrochloride salt of the compound of formula I at 20 mg/kg and 10 mg/kg, respectively, (the vehicle was normal saline). The positive drug group was subjected to intragastric administration with dexamethasone sodium phosphate with the dose of 0.05 mg/kg. The rats was administered once daily from Monday to Friday (i.e., consecutively five-day administration and then two-day interruption). The dosing period was two weeks.

[0094] 3. Detection and Results

[0095] After day 28 of the experiment, the blood was collected from the abdominal aorta of the rats in each group, and the serum was separated. The tumor necrosis factor-α (TNF-α) and the interleukin-1β (IL-1β) were detected by enzyme-linked immunosorbent assay, and the specific method was carried out according to the specifications of the kit. Statistical analysis was performed using SPSS 20.0 statistical software.

[0096] Determination of the rat paw swelling degree: before the primary immunization (d0), every 7 d (d7, d14) after the primary immunization and every 4 d after treatment, the thickness of the right hind paw of the rats was measured with a vernier caliper at the same position. Statistical analysis was performed using SPSS 20.0 statistical software.

[0097] After the animal blood collection was finished, ankle joints were fixed in 10% formalin. The ankle joints were subjected to decalcification by 10% acetic acid and then were subjected to routine paraffin embedding, slicing and HE staining. The slices were observed by pathology professionals, and then the damage degree of joint synovial tissue, joint bones and cartilage tissue was observed and scored for comparison.

[0098] Observation of lesions and scoring: 1. whether the joint synovial cells are proliferated or eroded (denatured, necrotic), and whether the inflammatory cells infiltrate or not; 2. whether the pannus is formed (composed of blood vessels and fibroblasts); 3. whether the bony erosion (joint bone and cartilage) or hyperplasia exists; 5. whether the joint cavity has stenosis, disappearance and other lesions. The observation indexes were classified into 3 grades according to the severity of the lesions, which were respectively recorded as: 1 point for slight, 2 points for moderate, and 3 points for severe. 0 points means substantially normal. All scores were accumulated to obtain a total score, and the average score (mean±SD) of each animal in each group was calculated, and the higher the score, the more severe the lesion.

[0099] The results showed that the joint surface of the rats in the normal group was covered with a thin layer of hyaline cartilage, the cells were neatly arranged without degeneration or necrosis, and the joint cavity was clear. The joint capsule was composed of loose connective tissue, and the surface of the tissue was covered with 1-4 layers of synovial cells. The cells have no degeneration, necrosis and hyperplasia. In the model group, the synovial cells were eroded, and there were different degrees of inflammatory cell infiltration and pannus in the synovial tissue. The bone was eroded, and the joint cavity was stenosed. The lesions in the dexamethasone sodium phosphate group were significantly reduced as compared to those in the model group. The joints of the administration group were improved in different degrees, and were improved in a dose-dependent manner.

[0100] In addition, the results also showed that compared with the model group, each group containing the test drugs could reduce the contents of TNF-α and IL-1β in the serum of rats, and the paw swelling degree was significantly reduced. The joint inflammatory infiltration was significantly reduced, the synovium hyperplasia was significantly relieved, and the joint cavity stenosis was significantly improved. It showed that the drug of the present invention can block inflammatory reaction, improve joint function and delay the progress of rheumatoid arthritis, and thus can be used for preparing the anti-rheumatoid arthritis drugs.

EXAMPLE 3

[0101] 1. Laboratory Animals

[0102] 15 SPF healthy male rats from Wistar with the weight of 160-180 g and purchased from Shanghai Institute of Planned Parenthood Research, with Certificate No. 20180006022473 in an SPF feeding environment.

[0103] 2. Experimental Reagents and Preparation Thereof

[0104] (2.1) Experimental Reagents

[0105] Bovine type II collagen (CII): Chondrex Inc.

[0106] Complete Freund's adjuvant (CFA): Chondrex Inc., CFA contains Bacille Calmette-Guerin vaccine (BCG) (BCG=1 mg/mL)

[0107] (2.2) Experimental Drugs

[0108] Anlotinib hydrochloride (i.e., the dihydrochloride salt of the compound of formula I). Dexamethasone sodium phosphate injection with the specification of 1 mL:5 mg.

[0109] (2.3) Preparation of Modeling Agent

[0110] Preparation of CII/CFA (BCG=1 mg/mL) emulsion: the vehicle of collagen solution was 0.05 M acetic acid, and the concentration of the collagen was 2 mg/mL The emulsion was prepared by emulsification with CFA (BCG=1 mg/mL) in equal volume ratio in a three-way tube. The emulsification operation was performed on ice for 30 min, and the final concentration of collagen was 1 mg/mL

[0111] (2.4) Preparation of Drugs

[0112] Dexamethasone solution: the commercially available dexamethasone sodium phosphate injection was diluted with normal saline to a concentration of 0.01 mg/mL for later use.

[0113] Anlotinib solution: the anlotinib hydrochloride was dissolved with normal saline, and diluted to a concentration of 1 mg/mL for later use.

[0114] 3. The Specific Experimental Operations Comprise the Followings.

[0115] (3.1) Establishment of Rheumatoid Arthritis CIA Model

[0116] 15 rats were randomly divided into two groups according to the weight, blank control group (n=3) and modeling group (n=12).

[0117] Modeling group: on day 1 of the experiment, the primary immunization was performed. The animals in the modeling group were anesthetized with isoflurane. Four points were taken to inject the modeling agent on both sides of the spinal cord skin at the base of the tail of the rats. A total of 0.2 mL of CII/CFA molding emulsion was injected intradermally. On day 8 of the experiment, the second immunization was performed, and the operations were the same as those on day 1 of the experiment. Blank control group: equal volume of normal saline was injected.

[0118] After modeling was completed, the animals were normally observed and housed. After the hind limb joints of the animals were diseased, the animals were grouped according to the joint swelling degree and the joint score, wherein the scoring criteria were as follows:

[0119] clinical diagnostic scoring criteria for joint soft tissue swelling and joint mobility:

TABLE-US-00001 Scoring Clinical symptoms 0 normal 1 slight, but significant redness and swelling or near redness and swelling of the ankle joints or knee joints, spread to the tips of the fingers (no matter how many) 2 severe redness and swelling of the ankle joints or knee joints 3 severe redness and swelling of the entire soles of foot, including the tips of fingers 4 cannot bear a weight

[0120] (3.2) Animal Grouping and Administration

[0121] Animals in the blank control group (n=3) were all entered Group-1 and administered with normal saline by intragastric administration.

[0122] The animals in the modeling group were divided into 3 groups according to the joint swelling degree and the joint scores: group-2 (n=4, model group), group-3 (n=4, positive control group), and group-4 (n=4, test drug group).

[0123] After grouping, oral gavage administration was started according to the grouping requirements, and the administration volume was 10 mL/kg. The administration was carried out once daily for 14 days (from day 1 to day 14). On day 15, the animals were dissected after collection of relevant experimental data. Specific types and modes of administration are shown in the following table:

TABLE-US-00002 Number of Group animals Administration regimen Group-1 3 Blank control Administration of normal group saline Group-2 4 Model group Administration of normal saline Group-3 4 Positive control Administration of group dexamethasone solution (0.1 mg/kg) Group-4 4 Test drug group Administration of anlotinib solution (10 mg/kg)

[0124] (3.3) Monitoring Items

[0125] 1) Weight Monitoring

[0126] During the experiment, the weight of the rats in each group was measured once daily from the day of grouping and administration (day 1) to day 15.

[0127] 2) Hind Limb Joint Size Monitoring

[0128] During the experiment, the width and thickness of the hind limb ankle joints of the rats in each group were measured with a vernier caliper on day 1, day 4, day 8, day 11 and day 15.

[0129] 3) Hind Limb Joint Lesion Scoring

[0130] During the experiment, hind limb ankle joints of the rats in each group were scored for joint soft tissue swelling and joint mobility on day 1, day 4, day 8, day 11 and day 15.

[0131] 4) Hind Limb Joint Weight Determination

[0132] On day 8, two rats in each group were dissected, and the weight of the hind limb joints was measured. The remaining animals were subjected to normal experiments according to the grouping schedule. On day 15, the remaining animals in each group were dissected, and the weight of the hind limb joints was measured.

[0133] 5) Joints Photographing

[0134] One rat in each group was selected, and the hind limb ankle joints thereof were photographed on day 4, day 8, day 11 and day 15.

[0135] 4. Experimental Results

[0136] 1) Effect of Drugs on the Weight of Rats

[0137] FIG. 1 shows the change (mean±sem) in the weight of the rats in each group over time during the experiment (note: model group vs. blank control group *P<0.05, **P<0.01, ***P<0.001). As shown in FIG. 1, after the onset of arthritis, the rats began to lose weight due to the immobility and loss of appetite caused by pain. In the positive control group, the rats had significant weight loss at the early stage, which was caused by the side effect of the hormone drug dexamethasone. After treatment with the drugs, the weight of the rats in the positive control group and the test drug group began to regain from day 7.

[0138] 2) Effect of Drugs on the Size of Hind Limb Joints of Rats

[0139] FIG. 2 shows the change (mean±sem) in the width of the hind limb ankle joints of the rats in each group over time during the experiment (note: model group vs. blank control group *P<0.05, ***P<0.001; positive control group vs. model group .sup.#P<0.05; test drug group vs. model group .sup.+P<0.05, .sup.+P<0.01). FIG. 3 shows the change (mean±sem) in the thickness of the hind limb ankle joints of the rats in each group over time during the experiment (note: model group vs. blank control group **P<0.01, ***P<0.001; positive control group vs. model group .sup.#P<0.05; test drug group vs. model group .sup.++P<0.01).

[0140] As shown in FIGS. 2 and 3, after treatment with the positive control drug, the width of the ankle joints of the rats in the positive control group was lower than that of the model group on day 15 (P<0.05), and the thickness of the ankle joints of the rats in the positive control group was lower than that of the model group on day 8 (P<0.05); after the treatment with the test drug, the width of the hind limb ankle joints of the rats in the test drug group was lower than that of the model group on day 4 (P<0.01), day 8 (P<0.01) and day 15 (P<0.05), and the thickness of the hind limb ankle joints of the rats in the test drug group was lower than that of the model group on day 4 and day 8 (P<0.01). It can be seen that the joint swelling degree of the positive control group and the test drug group has been improved to a certain extent as compared to that of the model group.

[0141] 3) Effect of Drugs on the Lesion Scores of Hind Limb Joints of Rats

[0142] FIG. 4 shows the change (mean±sem) in the lesion scores of the hind limb joints of the rats in each group over time during the experiment (note: model group vs. blank control group **P<0.01, ***P<0.001; positive control group vs. model group .sup.#P<0.05, .sup.##P<0.01; test drug group vs. model group .sup.+P<0.05, .sup.++P<0.01). As shown in FIG. 4, the scores of the hind limb ankle joints of the animals in the model group were increased as compared to those of the blank control group on day 4, day 8, day 11 and day 15, and the scores of the model group were significantly different as compared to the blank control group (P<0.001). The scores of the ankle joints in the positive control group were lower than those of the model group on day 8 (P<0.01), day 11 (P<0.05) and day 15 (P<0.05). The scores of the hind limb ankle joints in the test drug group were lower than those of the model group on day 4 (P<0.01), day 8 (P<0.01), day 11 (P<0.01) and day 15 (P<0.01).

[0143] 4) Effect of Drugs on the Weight of Hind Limb Joints of Rats

[0144] FIG. 5 shows the weight (mean±sem) of the hind limb joints of the rats in each group on day 8 (note: model group vs. blank control group ***P<0.001; positive control group vs. model group .sup.#P<0.05; test drug group vs. model group .sup.++P<0.01). As shown in FIG. 5, the weight of the hind limb joints of the rats in the positive control group was lower than that of the model group on day 8 (P<0.05), and the weight of the hind limb joints of the rats in the test drug group was also lower than that of the model group on day 8 (P<0.01).

[0145] 5) Effect of Drugs on the Size of Hind Limb Joints of Rats

[0146] As shown in FIGS. 6 and 7, during the experiment, the hind limb joints in the model group were significantly red and swollen, and the joint swelling degree of the positive control group and the test drug group was improved to a certain extent as compared to that of the model group.

[0147] In conclusion, the test drug showed significant drug efficacy on the rat rheumatoid arthritis model. It can improve joint function and delay the progress of rheumatoid arthritis.