TLR7 AGONIST FOR TREATING COLORECTAL CANCER AND PHARMACEUTICAL COMBINATION THEREOF

Abstract

Disclosed are a compound of formula I as a toll-like receptor 7 (TLR7) agonist or a pharmaceutically acceptable salt thereof for treating colorectal cancer, a pharmaceutical combination for treating colorectal cancer comprising the TLR7 agonist and a tyrosine kinase inhibitor, and the use of the compound of formula I or the pharmaceutically acceptable salt thereof and the pharmaceutical combination for treating colorectal cancer.

##STR00001##

Claims

1. A method for treating colorectal cancer, comprising administering to a subject in need thereof an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof: ##STR00059## wherein, L.sub.1 and L.sub.2 are each independently selected from the group consisting of —O—, —CH.sub.2—, —S—, —NH—, —NHC(═O)—, —C(═O)—, —C(═O)NH—, —S(═O)—, —S(═O).sub.2—, —NHS(═O).sub.2— and —S(═O).sub.2NH—, wherein the —CH.sub.2—, —NH—, —NHC(═O)—, —C(═O)NH—, —NHS(═O).sub.2— or —S(═O).sub.2NH— is optionally substituted with one or more R.sub.4; R.sub.1 is selected from the group consisting of hydrogen, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, 3-10 membered aryl and 3-10 membered heteroaryl, wherein the C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, 3-10 membered aryl and 3-10 membered heteroaryl are optionally substituted with one or more R.sub.4; R.sub.2 is selected from the group consisting of hydrogen, halogen, cyano, hydroxyl, sulfydryl, amino, —COOH, —CONH.sub.2, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, 3-10 membered aryl and 3-10 membered heteroaryl, wherein the hydroxyl, sulfydryl, amino, —COOH, —CONH.sub.2, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, 3-10 membered aryl and 3-10 membered heteroaryl are optionally substituted with one or more R.sub.4; B is selected from the group consisting of C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, 3-10 membered aryl and 3-10 membered heteroaryl; L.sub.3 is selected from the group consisting of a bond, C.sub.0-6 alkylene, imino, —O—, —S—, —S(═O)— and —S(═O).sub.2—, wherein the C.sub.0-6 alkylene and imino are optionally substituted with one or more R.sub.4; R.sub.3 is selected from the group consisting of hydrogen, amino, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, 3-10 membered aryl and 3-10 membered heteroaryl, wherein the amino, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 cyclohydrocarbyl, 3-10 membered heterocyclohydrocarbyl, 3-10 membered aryl and 3-10 membered heteroaryl are optionally substituted with one or more R.sub.4, or R.sub.3 and L.sub.3, together with an adjacent atom on ring B, form a saturated or unsaturated 5-8 membered ring optionally substituted with one or more R.sub.4; n is 0, 1, 2, 3, 4 or 5; and each R.sub.4 is independently selected from the group consisting of halogen, cyano, —R, —OR, ═O, —SR, —NR.sub.2, ═NR, —C(halogen).sub.3, —CR(halogen).sub.2, —CR.sub.2(halogen), —OCN, —SCN, —N═C═O, —NCS, —NO, —NO.sub.2, —NRC(═O)R, —NRC(═O)OR, —NRC(═O)NRR, —C(═O)NRR, —C(═O)OR, —OC(═O)NRR, —OC(═O)OR, —C(═O)R, —S(═O).sub.2OR, —S(═O).sub.2R, —OS(═O).sub.2OR, —S(═O).sub.2NRR, —S(═O)R, —NRS(═O).sub.2R, —NRS(═O).sub.2NRR, —NRS(═O).sub.2OR, —OP(═O)(OR).sub.2, —P(═O)(OR).sub.2, —C(═O)R, —C(═S)R, —C(═O)OR, —C(═S)OR, —C(═O)SR, —C(═S)SR, —C(═O)NRR, —C(═S)NRR, —C(═NR)NRR and —NRC(═NR)NRR; each R is independently selected from the group consisting of H, C.sub.1-8 alkyl, C.sub.3-8 cyclohydrocarbyl, 3-8 membered heterocyclohydrocarbyl, 3-8 membered aryl, 3-8 membered heteroaryl, 3-8 membered arylalkyl and 3-8 membered heteroarylalkyl; and when L.sub.1 is —CH.sub.2— or —NH—, R.sub.3 is not H.

2. The method according to claim 1, wherein L.sub.1 and L.sub.2 are each independently selected from the group consisting of —O—, —CH.sub.2—, —S—, —NH—, —C(═O)—, —S(═O)— and —S(═O).sub.2—, wherein the —CH.sub.2— and —NH— are optionally substituted with one or more R.sub.4.

3. The method according to claim 1, wherein R.sub.1 is selected from the group consisting of hydrogen, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cyclohydrocarbyl, 3-6 membered heterocyclohydrocarbyl, 3-6 membered aryl and 3-6 membered heteroaryl, wherein the C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cyclohydrocarbyl, 3-6 membered heterocyclohydrocarbyl, 3-6 membered aryl and 3-6 membered heteroaryl are optionally substituted with one or more R.sub.4.

4. The method according to claim 1, wherein R.sub.2 is selected from the group consisting of hydrogen, halogen, cyano, hydroxyl, sulfydryl, amino, —COOH, —CONH.sub.2, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cyclohydrocarbyl, 3-6 membered heterocyclohydrocarbyl, 3-6 membered aryl and 3-6 membered heteroaryl, wherein the hydroxyl, sulfydryl, amino, —COOH, —CONH.sub.2, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cyclohydrocarbyl, 3-6 membered heterocyclohydrocarbyl, 3-6 membered aryl and 3-6 membered heteroaryl are optionally substituted with one or more R.sub.4.

5. The method according to claim 1, wherein B is selected from the group consisting of 3-10 membered aryl and 3-10 membered heteroaryl.

6. The method according to claim 1, wherein L.sub.3 is selected from the group consisting of a bond and C.sub.0-6 alkylene, wherein the C.sub.0-6 alkylene is optionally substituted with one or more R.sub.4.

7. The method according to claim 1, wherein R.sub.3 is selected from the group consisting of hydrogen, amino, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-8 cyclohydrocarbyl, 3-8 membered heterocyclohydrocarbyl, 3-8 membered aryl and 3-8 membered heteroaryl, wherein the amino, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-8 cyclohydrocarbyl, 3-8 membered heterocyclohydrocarbyl, 3-8 membered aryl and 3-8 membered heteroaryl are optionally substituted with one or more R.sub.4; or R.sub.3 and L.sub.3, together with an adjacent atom on ring B, form a saturated or unsaturated 5-8 membered ring optionally substituted with one or more R.sub.4.

8. The method according to claim 1, wherein R.sub.4 is selected from the group consisting of halogen, cyano, —R, —OR, ═O, —SR, —NR.sub.2, ═NR, —C(halogen).sub.3, —CR(halogen).sub.2, —CR.sub.2(halogen), —OCN, —SCN, —N═C═O, —NCS, —NO, —NO.sub.2, —NRC(═O)R, —C(═O)NRR, —C(═O)OR, —OC(═O)NRR, —C(═O)R, —S(═O).sub.2OR, —S(═O).sub.2R, —OS(═O).sub.2OR, —S(═O).sub.2NRR, —S(═O)R, —NRS(═O).sub.2R, —C(═O)R, —C(═O)OR and —C(═O)NRR; and each R is independently selected from the group consisting of H, C.sub.1-8 alkyl, C.sub.3-8 cyclohydrocarbyl, 3-8 membered heterocyclohydrocarbyl, 3-8 membered aryl, 3-8 membered heteroaryl, 3-8 membered arylalkyl and 3-8 membered heteroarylalkyl.

9. The method according to claim 1, wherein the compound of formula I is selected from: 2-butoxy-7-(3-((4-methylpiperazin-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(3-(morpholinomethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 7-(3-(aminomethyl)benzyl)-2-butoxy-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(3-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((3,3-difluoropyrrolidin-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((3-fluoropyrrolidin-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 1-(4-((4-amino-2-butoxy-5H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl)benzyl)pyrrolidin-3-ol, 2-butoxy-7-(4-(piperidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-(morpholinomethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((4-methylpiperazin-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((dimethylamino)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((diethylamino)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((dipropylamino)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 7-(4-(azetidin-1-ylmethyl)benzyl)-2-butoxy-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((3-methoxyazetidin-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((4-methyl-1,4-diazepan-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((2,6-dimethylmorpholino)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 7-(4-(((1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl)methyl)benzyl)-2-butoxy-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((4-methoxypiperidin-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-((4-isopropylpiperazin-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-((6-(pyrrolidin-1-ylmethyl)pyridin-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(3-(2-(pyrrolidin-1-yl)ethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-(1-(pyrrolidin-1-yl)ethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-(1-methylpiperidin-4-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-(4-(1-methylpyrrolidin-2-yl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 1-(4-((4-amino-2-butoxy-5H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl)phenyl)-4-methylpiperazin-2-one, 7-benzyl-2-(2-methoxyethoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-(2-methoxyethoxy)-7-((6-methylpyridin-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 7-((5-chloropyridin-2-yl)methyl)-2-(2-methoxyethoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-(2-methoxyethoxy)-7-((6-(pyrrolidin-1-ylmethyl)pyridin-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 1-(4-((4-amino-2-(2-methoxyethoxy)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl)phenyl)-4-methylpiperazin-2-one, 2-butoxy-7-((5-(pyrrolidin-1-ylmethyl)pyridin-2-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 4-amino-2-butoxy-7-((6-(pyrrolidin-1-ylmethyl)pyridin-3-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-6-carbonitrile, 4-amino-2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-6-carbonitrile, 4-amino-2-butoxy-7-(4-(morpholinomethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-6-carbonitrile, 4-amino-2-butoxy-7-(4-((4-methylpiperazin-1-yl)methyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-6-carbonitrile, 4-amino-2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-6-carboxamide, 2-butoxy-7-((1,2,3,4-tetrahydroisoquinolin-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-((2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-((2-ethyl-1,2,3,4-tetrahydroisoquinolin-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-((2-isopropyl-1,2,3,4-tetrahydroisoquinolin-7-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-((1,2,3,4-tetrahydroisoquinolin-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-((2-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, 2-butoxy-7-((2-ethyl-1,2,3,4-tetrahydroisoquinolin-6-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine, or 2-butoxy-7-((2-(pyrrolidin-1-ylmethyl)thiazol-5-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine; or a pharmaceutically acceptable salt thereof.

10. (canceled)

11. (canceled)

12. A method for treating colorectal cancer, comprising administering to a subject in need thereof an effective amount of a pharmaceutical combination comprising one or more TLR7 agonists and anlotinib.

13. (canceled)

14. The method according to claim 12, wherein the TLR7 agonist is selected from one or more of imiquimod, GSK-2245035, resiquimod, vesatolimod, telratolimod, TMX-202, DSP-0509, RG-7854, loxoribine and the compound of formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1-9, or the TLR7 agonist is selected from one or more of GSK-2245035, vesatolimod, and 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine or 2-butoxy-7-((2-(pyrrolidin-1-ylmethyl)thiazol-5-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine or a pharmaceutically acceptable salt thereof, or the TLR7 agonist is selected from 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine or 2-butoxy-7-((2-(pyrrolidin-1-ylmethyl)thiazol-5-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine or a pharmaceutically acceptable salt thereof, or the TLR7 agonist is selected from 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine or a pharmaceutically acceptable salt thereof.

15. The method according to claim 12, wherein the pharmaceutical combination is selected from the group consisting of a fixed combination and a non-fixed combination.

16. The method according to claim 1, wherein the colorectal cancer is selected from the group consisting of colon cancer and rectal cancer.

17. The method according to claim 12, wherein the colorectal cancer is selected from the group consisting of colon cancer and rectal cancer.

18. The method according to claim 12, wherein anlotinib is anlotinib dihydrochloride.

19. The method according to claim 1, further comprising administering to the subject anlotinib or a pharmaceutically acceptable salt thereof.

20. The method according to claim 14, wherein the TLR7 agonist is administered in a dosage of 0.0001 mg/kg/d to 20 mg/kg/d, and anlotinib or a pharmaceutically acceptable salt thereof is administered in a unit dosage of 6 mg/d to 16 mg/d.

21. The method according to claim 12, wherein the TLR7 agonist and anlotinib are administered separately.

22. The method according to claim 15, wherein the fixed combination is in the form of a solid pharmaceutical composition, and the non-fixed combination are each in the form of a solid pharmaceutical composition.

23. The method according to claim 14, wherein the TLR7 agonist is selected from 2-butoxy-7-(4-(pyrrolidin-1-ylmethyl)benzyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine or a pharmaceutically acceptable salt thereof.

Description

DETAILED DESCRIPTION

[0145] For clarity, the present application is further described with the following examples, which are, however, not intended to limit the scope of the present application. All reagents used in the present application are commercially available and can be used without further purification.

[0146] A method for preparing the compound of formula I of the present application and its in vitro binding activity to a toll-like receptor 7 can be seen in WO2016023511 and WO2017076346.

Example 1 Anti-Tumor Experiment in MC-38 Mouse Model

[0147] In Example 1, the TLR7 agonist is selected from

##STR00058##

and anlotinib is selected fromanlotinib dihydrochloride.

[0148] The MC-38 is a colon cancer cell of mouse from Jiangsu Cyto Biotechnology Co., Ltd.

[0149] Subcutaneously grafted tumor cells of MC-38 colon cancer (concentration: 2×10.sup.6/mL×0.2 mL/mouse) were inoculated into female C57BL/6 mice at the right side armpit (the site to be inoculated was shaved before inoculating) in a sterile environment, and passaged using a block insertion method. The C57BL/6 mice were 16-18 g in weight, and the breeding environment was SPF level. The diameter of the C57BL/6 xenograft tumor was measured using a vernier caliper, and the mice were randomized into 4 groups when the tumors grew to 100-300 mm.sup.3 (12 mice for control group and 6 mice each for other groups):

[0150] i. Control group (blank control);

[0151] ii. Monotherapy group: anlotinib, with normal saline as vehicle;

[0152] iii. Monotherapy group: TLR7 agonist, with ethanol+tween 80+normal saline (v:v:v=5:5:90) as vehicle;

[0153] iv. Combination therapy group: TLR7 agonist+anlotinib.

TABLE-US-00002 TABLE 1 Administration regimen Route of Adminis- Adminis- Dosage adminis- tration tration Group mg/kg/time tration interval period i. — i.g. qd 2 w ii.  1 i.g. qd 2 w iii. 20 i.g. tiw 2 w iv. 1 (Anlotinib) i.g. qd 2 w 20 (TLR7 agonist) i.g. tiw 2 w i.g.: intragastric administration; qd: administration once daily; tiw: thrice a week.

[0154] The administration method of the combination therapy group is consistent with that of the monotherapy group.

[0155] Weights and diameter of the tumor were measured every three days, and the behavior of mice was observed daily. After the experiment was completed, the tumors were removed, weighed and photographed.

[0156] The tumor volume and the tumor growth inhibition were calculated using the following formulas:

Tumor volume (TV)=(L×W.sup.2)/2.

Tumor growth inhibition (TGI)(%)=(1−tumor weight in treatment group/tumor weight in control group)×100%.

[0157] The results as shown in Table 2 indicate that the TLR7 agonist can inhibit the growth of MC-38 tumor, and its combined use with anlotinib dihydrochloride can also inhibit the growth of MC-38 tumor with a synergistic effect.

TABLE-US-00003 TABLE 2 Experiment results for subcutaneous xenograft tumor of MC-38 colon cancer in mice TV (mm.sup.3) TV (mm.sup.3) TV (mm.sup.3) on day 0 on day 12 on day 18 TGI Group Mean ± SD Mean ± SD Mean ± SD (%) i. 143.06 ± 35.90 717.08 ± 362.31 1325.98 ± 595.27  — ii. 143.28 ± 45.46 427.30 ± 135.63 900.32 ± 346.06 33 iii. 143.93 ± 47.74 427.88 ± 161.31 747.63 ± 347.13 39 iv. 143.76 ± 46.04 191.00 ± 88.72  415.74 ± 183.33 69

[0158] The TGI (%), as a value of efficacy, was substituted into the Jin zhengjun's formula q=Eab/(Ea+Eb−Ea×Eb), where Ea and Eb represent the efficacy of medicaments used alone, and Eab represents the efficacy of combined use. The obtained q value is more than 1.15, showing synergistic effect of combined use.