Analogs of pentamidine and uses therefor

Abstract

To provide safe and effective, non-toxic pentamidine analogs that exhibit increased organ targeting that may allow for oncological clinical development designed for specific types of cancer.SOLUTION: The present disclosure provides a group of aromatic (e.g., pyridinyl, pyrimidinyl, pyrazinyl, or phenyl) diamidine analogs and pharmaceutically acceptable salts that are useful for treating a proliferative disease. The proliferative disease may include solid cancer or blood cancer. Compositions, methods of synthesizing the same and methods for treating various cancer using the analogs are disclosed herein. The present disclosure also provides pharmaceutical formulations comprising at least one of the compounds with a pharmaceutically acceptable carrier, diluent or excipient therefor.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] Cross - reference to related applications This application claims priority to U.S. Provisional Application No. 62 / 782,351, filed on December 20, 2018, the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to compounds useful for the treatment or prevention of mammals, particularly for the treatment of cancer.

Background Art

[0003] Pentamidine, 1,5 - bis(4 - amidinophenoxy)pentane, has been used for medical purposes since 1937 and is included in the World Health Organization's list of essential medicines as an antiprotozoal / antifungal agent for treating various infectious diseases (e.g., African trypanosomiasis, leishmaniasis, babesionsis, and Pneumocystis carinii pneumonia). Although the exact mode of its drug effect has not yet been elucidated, pentamidine is known to preferentially bind to the DNA in the minor groove of the AT - rich domain, and it has been proposed to exhibit anticancer activity through its inhibitory effect on PRL (phosphatase of the regenerating liver family), endo - exonuclease activity, and the interaction between S100B and p53.

[0004] Despite the fact that pentamidine has been used as an active therapeutic compound for decades, many side effects have severely limited the use of this drug for parasitic infections. Most of the treatments using this compound require careful monitoring of adverse events and dose - response because they can cause diabetes and harmful effects on the central nervous system. Among its side effects in particular, patients undergoing pentamidine treatment generally show a transient increase in serum liver transaminases (e.g., ALT and AST liver damage markers), indicating liver damage. Due to these potentially harmful results on important organ(s), the development of this compound as an anticancer agent, which often requires increased dosages because it is used for microbial infections, is severely restricted.

[0005] Pentamidine can be administered intramuscularly (IM) or intravenously (IV). However, only IV administration is recommended for the treatment of infectious diseases. This is because the oral bioavailability of this compound is low. Some studies have shown that the toxicity side effects of this drug can be controlled when given by aerosol administration. However, this particular administration method is limited to the treatment of pneumonia. Various approaches, such as pentamidine prodrugs, have been taken to overcome the drawbacks of this compound in oral bioavailability, but pentamidine analogs that provide safe and effective exposure at therapeutic levels, particularly via orally administered reduced toxicity, have not been reported to date.

[0006] Given the toxic side effects of pentamidine, there is an urgent need for safe, effective, and non-toxic pentamidine analogs that show increased organ targeting that could enable oncological clinical development designed for specific types of cancer.

Brief Description of the Drawings

[0007]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Summary of the Invention

[0008] The present disclosure relates to a group of aromatic (e.g., pyridinyl, pyrimidinyl, pyrazinyl, or phenyl) diamidine analogs and pharmaceutically acceptable salts useful for the treatment of proliferative diseases. Proliferative diseases may include solid cancers or blood cancers. Compositions, methods for synthesizing them, and methods for treating various cancers using the analogs are disclosed herein. The present disclosure also provides a pharmaceutical formulation comprising at least one of the compounds having a pharmaceutically acceptable carrier, diluent, or excipient therefor.

[0009] The present invention is based on the discovery that pentamidine analogs are useful for treating various types of cancer including, but not limited to, liver cancer, lung cancer, colon cancer, bile duct cancer, kidney cancer, stomach cancer, melanoma, ovarian cancer, breast cancer, and pancreatic cancer. These aromatic diamidine compounds exhibit similar or increased cytotoxicity against cancer cells compared to pentamidine, and also show improved pharmacokinetics and pharmacodynamics in the liver, as well as a significant improvement in oral bioavailability, and are significantly safer than pentamidine or other standard therapeutic molecules. In summary, due to these properties, the compounds of the present invention are highly desirable for clinical development for cancer treatment.

[0010] In one aspect, the present invention relates to a composition of a pentamidine analog having the formula (A)

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chem.

[0011] In some variants of formula (A),

Chem.

[0012] In some embodiments, Y1 -Y 10 At least one of which is N. In some embodiments, Y 1 -Y 5 The ring formed by is Y 6 -Y 10 is different from the ring formed by. In some embodiments, Y 1 -Y 10 The two amidine-substituted rings of are different so that the compound is not symmetric.

[0013] Therefore, formula (A) is the following compound 6,6'-(Pentane-1,5-diylbis(oxy))dinicotinimidamide; 5,5'-(Pentane-1,5-diylbis(oxy))dipicolimidamide; 4,4'-(Pentane-1,5-diylbis(oxy))dipicolimidamide; 6,6'-(Pentane-1,5-diylbis(oxy))dipicolimidamide; 6,6'-(Cyclohexane-1,3-diylbis(oxy))dinicotinimidamide; and 5,5'-(1,4-Phenylenebis(oxy))dipicolimidamide, or a pharmaceutically acceptable salt thereof is understood to be excluded.

[0014] In some embodiments, the present invention relates to a composition of pentamidine analogs having formula (I):

Chemical formula

Chemical formula

[0015] In some embodiments, Y 1 -Y 10 at least one of is N. In some embodiments, Y 1 -Y 5 The ring formed by is different from the ring formed by Y 6 -Y 10 In some embodiments, Y 1 -Y 10 The two amidine-substituted rings of are different such that the compound is not symmetric.

[0016] In one embodiment, m is 1 and n is 1. In another embodiment, m is 1 and n is 0. In another embodiment, m is 0 and n is 1. In another embodiment, m is 1 and n is 2. In another embodiment, m is 2 and n is 1. In one embodiment, m is 2 and n is 2. In another embodiment, m is 0 and n is 0.

[0017] In one embodiment, Z 1 or Z 2 is independently O and may be substituted. In another embodiment, Z 1 or Z 2 is independently S and may be substituted. In yet another embodiment, Z 1 or Z 2 is independently NR 3 wherein R 3 is hydrogen. In one embodiment, Z 1 or Z 2 is independently NR 3 wherein R 3 is alkyl, cycloalkyl, aryl, or heteroaryl. In another embodiment, Z 1 or Z 2 is independently NR 3 or CR 5 R 6 In another embodiment, Z 1 is NR 3and wherein R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, and Z 2 is CR 5 R 6 and wherein R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring.

[0018] In one embodiment, the amidine is independently bonded by Y 3 and Y 8 In another embodiment, the amidine is independently bonded by Y 3 and Y 7 In yet another embodiment, the amidine is independently bonded by Y 2 and Y 7 In yet another embodiment, the amidine is independently bonded by Y 2 and Y 7

[0019] In one embodiment, Y 1、2、4、5、6、8 is CR 7 (e.g., -CH), Y 2 is N, and Y 3 and Y 7 are bonded to the amidine. In another embodiment, Y 1、4、5、6、及び7 is -CH, Y 2 is N, Y 3 and Y 8 are CR 7 wherein R 7 is the amidine. In another embodiment, Y 1、4、5、6、及び8 is -CH, Y 3 is N, Y 2 and Y 7 are CR 7 wherein R 7 is the amidine. In another embodiment, Y 1、4、5、6、及び8 is -CH, Y 3 is N, Y 2 and Y​7 is CR 7 wherein R 7 is amidine, m is 1, and n is 0. In another embodiment, Y 1、4、5、及び6 is -CH, Y 3 and Y 8 are N, Y 2 and Y 7 are CR 7 wherein R 7 is amidine, m is 1, and n is 0.

[0020] In one embodiment, R 1 and R 2 are independently hydrogen. In another embodiment, R 1 is R 2 which together with R forms a saturated, unsaturated, or partially unsaturated 3- to 9-membered cyclic group (e.g.,

Chemical formula

Mode for Carrying Out the Invention

[0021] Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings commonly understood by those of ordinary skill in the art. The meanings and scopes of the terms should be clear, but if potential ambiguity arises, the definitions provided herein shall take precedence over dictionary or external definitions. The term "including", as well as other forms such as "include" and "included", is not limiting.

[0022] As used herein, "a" or "an" means "at least one" or "one or more".

[0023] As used herein, "or" means "and / or".

[0024] As used herein, the term "alkyl" refers to saturated hydrocarbon groups in linear, branched, or cyclic arrangements, or any combination thereof, and particularly contemplated alkyl groups include those having 10 or fewer carbon atoms, particularly those having 1 to 6 or fewer carbon atoms, and lower alkyl groups having 1 to 4 carbon atoms. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, pentyl, isopentyl, hexyl, cyclopropylmethyl, and the like. The alkyl group may be unsubstituted or substituted to the extent such substitution is chemically feasible. Typical substituents include halo, =O, =N-CN, =N-OR a 、=NR a 、-OR a 、-NR a 2 、-SR a 、-SO 2 R a 、-SO 2 NR a 2 、-NR a SO 2 R a 、-NR a CONR a 2 、-NR a COOR a 、-NR a COR a 、-NO 2 、-CN、-COOR a 、-CONR a 2 、-OOCR a 、-COR a 、and R a are included, but are not limited thereto. Here, each R a is independently H, C 1 -C 8 alkyl, C2 -C 8 Heteroalkyl, C 3 -C 8 Heterocyclyl, C 4 -C 10 Heterocycloalkyl, C 1 -C 8 Acyl, C 2 -C 8 Heteroacyl, C 2 -C 8 Alkenyl, C 2 -C 8 Heteroalkenyl, C 2 -C 8 Alkynyl, C 2 -C 8 Heteroalkynyl, C 6 -C 10 Aryl, or C 5 C 10 is heteroaryl, and each R a is halo, =O, =N-CN, =N-OR b 、=NR b 、-OR b 、-NR b 2 、-SR b 、-SO 2 R b 、-SO 2 NR b 2 、-NR b SO 2 R b 、-NR b CONR b 2 、-NR b COOR b 、-NR b COR b 、-NO 2 、-N、-COOR b 、-CONR b 2 、-OOCR b 、-COR b 、and R b may be substituted, where each R bは is independently H, C 1 -C 8 alkyl, C 2 -C 8 heteroalkyl, C3 -C 8 Heterocyclyl, C 4 -C 10 Heterocyclyl, C 1 -C 8 Acyl, C 2 -C 8 Heteroacyl, C 2 -C 8 Alkenyl, C 2 -C 8 Heteroalkenyl, C 2 -C 8 Alkynyl, C 2 -C 8 Heteroalkynyl, C 6 -C 10 Aryl or C 5 -C 10 is heteroaryl. The alkyl, alkenyl, and alkynyl groups are C 1 -C 8 Acyl, C 2 -C 8 Heteroacyl, C 6 -C 10 Aryl, or C 5 -C 10 can also be substituted with heteroaryl, and each can be substituted with a substituent appropriate for the particular group. When the substituent contains two R a or R b groups on the same or adjacent atoms (e.g., -NR b 2 , or NR b -C(O)R b ), the two R a or R b groups can optionally combine with the atoms of the attached substituent to form a ring having 5 to 8 ring members, and this ring can be substituted as permitted for R a or R b itself and can include additional heteroatoms (N, O, or S) as ring members.

[0025] As used herein, the term "alkenyl" refers to a hydrocarbon chain having at least two carbon atoms and at least one carbon-carbon double bond, and includes straight-chain, branched, or cyclic alkenyl groups having from 2 to 10 carbon atoms. Non-limiting examples of "alkenyl" include ethenyl, propenyl, butenyl, pentenyl, and cyclic alkenyl groups. The alkenyl group can be unsubstituted or substituted with one or more suitable substituents.

[0026] As used herein, the term "alkynyl" refers to unbranched and branched hydrocarbon moieties having at least two (preferably three) carbon atoms and at least one carbon-carbon triple bond, and includes ethynyl, propynyl, butynyl, cyclopropyl ethynyl, and the like. The alkynyl group can be unsubstituted or substituted with one or more suitable substituents.

[0027] As used herein, the term "alkoxy" refers to the above alkyl group bonded through oxygen, examples of which include methoxy, ethoxy, propyloxy, isopropoxy, tert-butoxy, methoxyethoxy, benzyloxy, allyloxy, and the like. Also, alkoxy means polyethers such as -O-(CH 2 ) 2 -O-CH 3 . Alkoxy can be any hydrocarbon group connected through an oxygen atom, where the hydrocarbon moiety can have any number of carbon atoms, typically from 1 to 10 carbon atoms, can further include double or triple bonds, and can include one or two oxygen, sulfur, or nitrogen atoms in the alkyl chain. The alkoxy group can be unsubstituted or substituted with one or more suitable substituents, such as aryl, heteroaryl, cycloalkyl, and / or heterocyclyl.

[0028] As used herein, the term "cycloalkyl" refers to a cyclic alkane in which the carbon atoms of the hydrocarbon form a ring, and includes monocyclic or polycyclic hydrocarbon ring groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, adamantyl, norpinanyl, decalinyl, norbornyl, housanyl, etc. Further, cycloalkyl can also include one or two double bonds that form a "cycloalkenyl" group (such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, norbornenyl, norbornadienyl, etc.). Cycloalkyl can also contain one or more heteroatoms and is called "cycloheteroalkyl", and can include, for example, piperazinyl piperidinyl, morpholinyl, thiomorpholinyl, oxanyl, dioxanyl (such as 1,4-dioxanyl), thianyl, dithianyl, hexahydro-1,3,5-triazinyl, trioxanyl, trithianyl, pyrrolidinyl, imidazolidinyl, pyranyl, tetrahydropyranyl, pyrazolidinyl, oxolanyl, oxazolidinyl, thiolanyl, thiazolidinyl, pyrrolinyl, pyrazolinyl, imidazolinyl, tetrahydrofuranyl, etc. The cycloalkyl or cycloheteroalkyl group can be unsubstituted or substituted with one or more suitable substituents.

[0029] As used herein, the term "amidine" or "Am" refers to a -CNH 2 NH group as shown in the following structure.

Chemical formula

[0030] The term "hetero", as used herein, refers to an atom of any element other than carbon or hydrogen. As used herein, the term "heteroatom" means nitrogen (N), oxygen (O), or sulfur (S).

[0031] As used herein, the terms "heterocyclic" or "heterocyclyl" include all restrictions of "cycloheteroalkyl" and "heteroaryl" groups to the extent chemically practicable. The terms "heterocyclic" or "heterocyclyl" refer to any compound in which a plurality of atoms form a ring via a plurality of covalent bonds, where the ring contains at least one atom other than a carbon atom as a ring member. The heterocyclic ring may be saturated, unsaturated, or partially unsaturated. The unsaturated heterocyclic ring may be an aromatic aryl. Non-limiting examples of heterocyclic rings include 3-, 4-, 5-, 6-, 7-, 8-, and 9-membered monocyclic rings containing one or more N, O, or S as non-carbon member(s) (plural possible), as follows. (1) Saturated 3-atom heterocyclic rings can be, for example, aziridinyl, diaziridinyl, oxiranyl, dioxiranyl, oxaziridinyl, thiiranyl, etc., and unsaturated 3-atom heterocyclic rings can be, for example, azirinyl, oxirenyl, thiirenyl, diazirinyl, etc.; (2) Saturated 4-atom heterocyclic rings can be, for example, azetidinyl, diazetidinyl, oxetanyl, dioxetanyl, thietanyl, dithietanyl, etc., and unsaturated 4-atom heterocyclic rings can be, for example, azetyl, diazetyl, oxetyl, dioxetyl, thietyl, dithietyl, etc.; (3) Saturated 5-atom heterocyclic rings can be, for example, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxolanyl, oxazolidinyl, thiolanyl, thiazolidinyl, etc., and unsaturated and partially unsaturated 5-atom heterocyclic rings can be, for example, pyrrolyl, pyrrolinyl, pyrazolyl, pyrazolinyl, imidazolyl, imidazolinyl, triazolyl, tetrazolyl, thiophenyl, thiazolyl, dithiazolyl, thiazolinyl, isothiazolyl, thiadiazolyl, furanyl, furazanyl, oxazolyl, isoxazolyl, oxazolyl, etc.; (4) Saturated 6-atom heterocyclic rings can be, for example, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, oxanyl, dioxanyl (e.g., 1,4-dioxane cyclohexane), thianyl, dithianyl, hexahydro-1,3,5-triazinyl, trioxanyl, trithianyl, etc., and unsaturated 6-atom heterocyclic rings can be, for example, pyridinyl, diazinyl (e.g., pyrimidinyl, or pyridazinyl), pyranyl, oxazinyl (e.g., 1,2-oxazinyl; 1,3-oxazinyl, or 1,(4 - oxazinyl), thiazinyl, 1,4 - dioxinyl, dithiinyl, triazinyl (e.g., 1,2,3 - triazinyl, 1,2,4 - triazinyl, or 1,3,5 - triazinyl), tetrazinyl, pentazinyl, thiopyranyl, etc.; (5) The saturated 7 - atom heterocyclic ring can be, for example, azepanyl, diazepanyl, oxepanyl, thiepanyl, etc., and the unsaturated 7 - atom heterocyclic ring can be, for example, azepinyl, diazepinyl, oxepinyl, thiepinyl, thiazepinyl, etc.; (6) The saturated 8 - atom heterocyclic ring can be, for example, azocanyl, oxocanyl, thiocanyl, etc., and the unsaturated 8 - atom heterocyclic ring can be, for example, azocinyl, oxocinyl, thiocinyl, etc.; (7) The saturated 9 - atom heterocyclic ring can be, for example, azonanyl, oxonanyl, thionanyl, etc., and the unsaturated 9 - atom heterocyclic ring can be, for example, azoninyl, oxoninyl, thioninyl, etc. Further contemplated heterocyclic rings can, for example, covalently bond to two atoms on a first non - heterocyclic ring group (e.g., phenyl) to condense with one or two heterocyclic rings (e.g., 1,4 - dioxanyl, 1,4 - dioxinyl, and tetrahydropyranyl), or covalently bond to two atoms on a first heterocyclic ring (e.g., pyrrolyl, imidazolyl, thiazolyl, pyrimidinyl, and pyridinyl) to condense with one or two non - heterocyclic rings or heterocyclic ring groups (e.g., 1,4 - dioxanyl, 1,4 - dioxinyl, and morpholinyl), and collectively, are referred to herein as "condensed heterocyclic ring" or "condensed heterocyclic ring moiety" or "heteroaryl - fused cycloheteroalkyl". The condensed heterocyclic ring can be, for example, a saturated or unsaturated (e.g., aromatic) bicyclic or tricyclic compound. Non - limiting examples of the condensed heterocyclic ring include dihydrobenzodioxinyl, dihydrodioxinopyridinyl, dihydrodioxinopyridazinyl, dihydrodioxinopyrimidinyl, dihydrodioxinopyrazinyl, dihydropyrrolopyridinyl, tetrahydronaph, tetrahydropyridopyridazinyl, tetrahydropyridopyrazinyl, tetrahydropyridopyrimidinyl, chromanyl, indolyl, purinyl, isoindolyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl, quinolidinyl, 1,8 - naphthyridinyl, pyrido[3,2 - d]pyrimidinyl, pyrido[4,3-d] pyrimidinyl, pyrido[3,4-b]pyrazinyl, pyrido[2,3-b]pyrazinyl, pteridinyl, acridinyl, cinnolinyl, phthalazinyl, benzimidazolyl, phenazinyl, phenoxazinyl, phenothiazinyl, phenoxathiinyl, benzazepinyl, benzodiazepinyl, benzofuranyl, dibenzofuranyl, isobenzofuranyl, benzothiophenyl, benzoxazinyl, quinolin-2(1H)-onyl, isoquinolin-1(2H)-onyl, indazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, dibenzazepinyl, dibenzoxepinyl, dibenzothiazepinyl, dibenzothiepinyl, carbazolyl, fluorenyl and the like. When the heterocyclic ring is aromatic, it can also be referred to as "heteroaryl" or "heteroaromatic" as further described below. The non-aromatic heterocyclic ring can be substituted with any group suitable for the substituents of the above alkyl groups.,

[0032] As used herein, the term "aryl" refers to an unsubstituted or substituted aromatic monocyclic or polycyclic group, which may further contain one or more non-carbon atoms. The term "aryl" also includes a non-aromatic carbocyclic ring or an aromatic ring fused to a heterocyclyl group having 1 to 7 heteroatoms. The term "aryl" can be used interchangeably with "aryl ring", "aromatic group", and "aromatic ring". An aryl group may generally contain 1 to 9 heteroatoms, which are generally called "heteroaryl". A heteroaryl group usually has 4 to 14 atoms, of which 1 to 9 are independently selected from the group consisting of N, O, and S. For example, in a 5- to 8-membered aromatic group, a heteroaryl group can contain 1 to 4 heteroatoms. An aryl or heteroaryl group can be unsubstituted or substituted with one or more suitable substituents.,

[0033] Aryl or heteroaryl can be a monocyclic or polycyclic (e.g., bicyclic) aromatic group. Typical aryl groups include, for example, phenyl and naphthalenyl. Typical heteroaryl groups include, for example, quinolinyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, thiophenyl, thiazolyl, dithiazolyl, thiazolinyl, isothiazolyl, thiadiazolyl, furanyl, furazanyl, oxazolyl, isoxazolyl, oxadiazolyl, pyridinyl, diazinyl (e.g., pyrazinyl, pyrimidinyl, or pyridazinyl), triazinyl (e.g., 1,2,3-triazinyl, 1,2,4-triazinyl, or 1,3,5-triazinyl), pyranyl, oxazinyl (e.g., 1,2-oxazinyl; 1,3-oxazinyl, or 1,4-oxazinyl), thiazinyl, dioxinyl, dithiinyl, triazinyl, tetrazinyl, pentazinyl, thiopyranyl, azepinyl, diazepinyl, oxepinyl, thiepinyl, thiazepinyl, azocinyl, oxocinyl, thiocinyl, azoninyl, oxoninyl, thioninyl, indolyl, indazolyl, purinyl, isoindolyl, quinolinyl, isoquinolinyl, quinoxalinyl, acridinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzimidazolyl, benzofuranyl, isobenzofuranyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, and the like. The polycyclic aryl or polycyclic heteroaryl group can be formed by condensing (i.e., covalently bonding) at least one carbon cyclic or heterocyclic group to two atoms on the first aryl or heteroaryl ring, and is thus called "condensed aryl" or "heteroaryl condensed cycloheteroalkyl".

[0034] As used herein, the term "heteroaryl-fused cycloheteroalkyl" refers to a heterocyclyl moiety consisting of a monocyclic heteroaryl group such as pyridinyl or furanyl fused to a cycloheteroalkyl group, wherein the heteroaryl and cycloheteroalkyl moieties are as defined herein. Exemplary heteroaryl-fused heterocycloalkyl groups include dihydrodioxinopyridinyl, dihydrodioxinopyridazinyl, dihydrodioxinopyrimidinyl, dihydrodioxinopyrazinyl, dihydrodioxinotriazinyl, dihydropyrrolopyridinyl, dihydrofuranylpyridinyl, and dioxolopyridinyl. The heteroaryl-fused heterocycloalkyl group can be attached to the remainder of the molecule by any available carbon or nitrogen atom.

[0035] Typical heteroaryl groups include 5- or 6-membered monocyclic aromatic groups such as pyridinyl, pyrimidyl, pyrazinyl, pyridazinyl, thienyl, furanyl, pyrrolyl, pyrazolyl, thiazolyl, oxazolyl, isothiazolyl, isoxazolyl, thiophenyl, triazolyl (1,2,4-triazolyl and 1,2,3-triazolyl), tetrazolyl, furazanyl, oxadiazolyl (1,2,5-oxadiazolyl and 1,2,3-oxadiazolyl), and imidazolyl, and fused bicyclic moieties formed by condensing one of the heterocyclic groups with either a phenyl ring or a heteroaromatic monocyclic group include indolyl, benzimidazolyl, indazolyl, benzotriazolyl, isoquinolyl, quinolyl, benzothiazolyl, benzofuranyl, pyrazolopyridinyl, pyrazolopyrimidyl, quinazolinyl, quinoxalinyl, cinnolinyl, imidazopyrimidinyl, and the like.

[0036] As used herein, the term "monocyclic" refers to an unsubstituted or substituted monocyclic structure. As used herein, the terms "polycyclic" and "bicyclic" refer to an unsubstituted or substituted polycyclic structure comprising at least two ring structures fused by any two adjacent atoms. A bicyclic ring can be an aryl or heteroaryl ring fused to an aromatic ring, or a non-aromatic carbocyclic ring such as cycloalkyl or cycloheteroalkyl. A bicyclic ring can also be a non-aromatic carbocyclic ring fused to another non-aromatic carbocyclic ring such as cycloalkyl or cycloheteroalkyl. Non-limiting examples of bicyclic rings include dihydrobenzodioxinyl, dihydrodioxinopyridinyl, dihydrodioxinopyridazinyl, dihydrodioxinopyrimidinyl, dihydrodioxinopyrazinyl, dihydropyrrolopyridinyl, tetrahydronaphthyridinyl, tetrahydropyridopyridazinyl, tetrahydropyridopyrazinyl, tetrahydropyridopyrimidinyl, chromanyl, decalinyl, purinyl, indolyl, isoindolyl, quinolyl, quinazolinyl, benzimidazolyl, imidazopyridinyl, cinnolinyl, phthalazinyl, imidazopyrimidinyl, and the like. Any monocyclic or fused bicyclic system having aromatic characteristics with respect to the overall electronic distribution of the ring system is included in this definition. It also includes bicyclic groups in which at least the ring directly bonded to the remainder of the molecule has aromatic characteristics.

[0037] When permitted, aryl and heteroaryl groups can be substituted. Suitable substituents include halo, R a , -OR a , -NR a 2 , -SR a , -SO 2 R a , -SO 2 , -SO a 2 , -NR a , -NR 2 SO a , -NR a , -NR a 2 , -NR a , -NR a , -NRa COR a 、 -CN, -COOR a 、 -CONR a 2 、 -OOCR a 、 -COR a 、 and -NO 2 is included, but not limited to these. Here, each R a is independently H, C 1 -C 8 alkyl, C 2 -C 8 heteroalkyl, C 3 -C 8 heterocyclyl, C 4 -C 10 heterocycloalkyl, C 1 -C 8 acyl, C 2 -C 8 heteroacyl, C 2 -C 8 alkenyl, C 2 -C 8 heteroalkenyl, C 2 -C 8 alkynyl, C 2 -C 8 heteroalkynyl, C 6 -C 10 aryl, or C 5 -C 10 heteroaryl, and each R a is halo, =O, =N-CN, =N-OR b 、 =NR b 、 -OR b 、 -NR b 2 、 -SR b 、 -SO 2 R b 、 -SO 2 NR b 2 、 -NR b SO 2 R b 、 -NR b CONR b 2 、 -NR b COOR b 、 -NR b COR b 、 -CN, -COORb , -CONR b 2 , -OOCR b , -COR b , and NO 2 may be replaced by, wherein each R b is independently H, C 1 -C 8 alkyl, C 2 -C 8 heteroalkyl, C 3 -C 8 heterocyclyl, C 4 -C 10 heterocycloalkyl, C 1 -C 8 acyl, C 2 -C 8 heteroacyl, C 2 -C 8 alkenyl, C 2 -C 8 heteroalkenyl, C 2 -C 8 alkynyl, C 2 -C 8 heteroalkynyl, C 6 -C 10 aryl, or C 5 -C 10 heteroaryl. Alkyl, alkenyl, and alkynyl groups may be substituted with C 1 -C 8 acyl, C 2 -C 8 heteroacyl, C 6 -C 10 aryl, or C 5 -C 10 heteroaryl, and each may be substituted with a substituent appropriate for the particular group. When a substituent contains two R a or R b groups on the same or adjacent atoms (e.g., -NR b 2 , or NR b -C(O)R b ), the two R a or R b groups may optionally combine with the atoms of the attached substituent to form a ring having 5 to 8 ring members, and this ring is an Ra or R b It may be replaced as permitted by itself and may include additional heteroatoms (N, O, or S) as ring members.

[0038] "Sulfonyl" means SO 2 -alkyl, SO 2 -substituted alkyl, SO 2 -alkenyl, SO 2 -substituted alkenyl, SO 2 -cycloalkyl, SO 2 -substituted cycloalkyl, SO 2 -cycloalkenyl, SO 2 -substituted cycloalkenyl, SO 2 -aryl, SO 2 -substituted aryl, SO 2 -heteroaryl, SO 2 -substituted heteroaryl, SO 2 -heterocyclic ring, and SO 2 -substituted heterocyclic group, and alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic ring, and substituted heterocyclic ring are each as defined herein.

[0039] As used herein, the term "acyl" when used without the modifier "substituted" refers to a -C(O)R group, where R is hydrogen, alkyl, aryl, halide, aralkyl or heteroaryl, and these terms are defined herein.

[0040] As used herein, the term "acyloxy" refers to a straight-chain or branched alkanoyl group having 1 to 6 carbon atoms such as formyl, acetyl, propanoyl, butyryl, valeryl, pivaloyl and hexanoyl, as well as an arylcarbonyl group described below, or a heteroarylcarbonyl group described below. The aryl moiety of the arylcarbonyl group means a group having 6 to 16 carbon atoms such as phenyl, biphenyl, naphthyl, or pyrenyl. The heteroaryl moiety of the heteroarylcarbonyl group contains at least one heteroatom from O, N, and S, such as pyridinyl, pyrimidyl, pyrrolyl, furyl, benzofuryl, thienyl, benzothienyl, imidazolyl, triazolyl, quinolyl, isoquinolyl, benzimidazolyl, thiazolyl, benzothiazolyl, oxazolyl, and indolyl.

[0041] As used herein, the term "carboxylic acid" refers to a -C(O)OH group.

[0042] As used herein, the term "ester" as used herein refers to a -C(O)O- group.

[0043] As used herein, the term "nitro" refers to -NO 2 and means.

[0044] As used herein, the term "cyano" means -CN.

[0045] As used herein, the term "azide" means related to a monovalent group containing -N 3 and means.

[0046] As used herein, the term "sulfhydryl" means thiol, -SH.

[0047] As used herein, the term "amine" refers to primary, secondary and tertiary amines, -R-NH 2、 -R-NH-R', and R-N-(R")R'.

[0048] As used herein, the term "amide" means a primary, secondary, and tertiary amide, -R-C(O)NH 2 、 -RC(O)NH-R', and R-C(O)NR'R".

[0049] As used herein, the term "carbonate" means an ester of carbonic acid having a group containing C(=O)(O-) 2 is meant.

[0050] As used herein, the term "carbamate" means a group containing NH 2 COOH.

[0051] As used herein, the term "hydroxyl" means -OH.

[0052] As used herein, the terms "halo", "halogen", and "halide" mean fluoro (-F), chloro (-Cl), bromo (-Br), and iodo (-I).

[0053] As used herein, the term "haloalkyl" refers to any alkyl in which one or more hydrogen atoms are replaced by one or more halogen atoms. Non-limiting examples of haloalkyl include

[0054] -CF 3 、 -CFH 2 、 -CF 2 H, etc.

[0055] As used herein, the term "arylalkyl" refers to any alkyl in which one or more hydrogen atoms are replaced by an aryl or heteroaryl group. Examples of arylalkyl include benzyl (C 6 H 5 CH 2 -), etc.

[0056] As used herein, the term "hydroxyalkyl" refers to any hydroxy derivative of an alkyl and includes any alkyl in which one or more hydrogen atoms are replaced by an -OH group.

[0057] The term "haloalkyl" refers to an alkyl group as described above having one or more hydrogen atoms on an alkyl group substituted with a halo group. Examples of such groups include, but are not limited to, fluoroalkyl groups such as fluoroethyl, difluoromethyl, trifluoromethyl, trifluoroethyl, and the like.

[0058] The term "haloalkoxy" refers to an alkyl - O - group in which one or more hydrogen atoms on the alkyl group are replaced by a halo group (e.g., -F, -Cl, -Br, and -I), and examples of such groups include groups such as trifluoromethoxy.

[0059] As used herein, the term "substituted" refers to the replacement of a hydrogen atom of an unsubstituted group with a functional group, and the particularly contemplated functional groups include nucleophilic groups (e.g., -NH 2 , -OH, -SH, -CN, etc.), electrophilic groups (e.g., C(O)OR, C(X)OH, etc.), polar groups (e.g., -OH), non - polar groups (e.g., heterocyclic ring, aryl, alkyl, alkenyl, alkynyl, etc.), ionic groups (e.g., -NH 3 + ), and halogen (e.g., -F, -Cl), NHCOR, NHCONH 2 , OCH 2 COOH, OCH 2 CONH 2 , OCH 2 CONHR, NHCH 2 COOH, NHCH 2 CONH 2 , NHSO 2 R, OCH 2 heterocyclic ring, PO 3 H, SO 3H, amino acids, and all chemically reasonable combinations thereof are included. Further, the term "substituted" also includes multiple degrees of substitution, and when multiple substituents are disclosed or claimed, the substituted compound may be independently substituted by one or more of the disclosed or claimed substituent moieties.

[0060] Unless otherwise specified, the nomenclature of substituents not explicitly defined herein is obtained by naming the functional group adjacent to the point of attachment towards the point of attachment following the terminal portion of the functional group. For example, the substituent "alkylaryl-oxycarbonyl" refers to the (alkyl)-(aryl)-O-C(O)- group.

[0061] With respect to any of the groups disclosed herein that contain one or more substituents, it is understood that such groups do not include any substitution or substitution pattern that is sterically unrealistic and / or synthetically infeasible. Further, the compounds of the present invention include all stereochemical isomers resulting from the substitution of these compounds.

[0062] In addition to the disclosure herein, in certain embodiments, the group to be substituted has one substituent, one or two substituents, one, two, or three substituents, one, two, three, or four substituents, or four substituents.

[0063] As used herein, the term "administering" or "administration" of a subject compound refers to providing the compound of the present invention to a subject in need of treatment.

[0064] As used herein, the term "acceptable" with respect to a formulation, composition, or component used herein means that it does not have a continuing harmful effect on the general health of the subject being treated.

[0065] The term "about" when referring to a numerical value or range of numerical values means that the recited numerical value or range of numerical values is an approximation within experimental variability (or within statistical experimental error), and thus the numerical value or range of numerical values can vary, for example, by 1% to 10% of the specified numerical value or range of numerical values.

[0066] As used herein, the term "carrier" refers to a compound or agent that facilitates the incorporation of the compounds described herein into cells or tissues.

[0067] As used herein, the terms "comprise", "have", and "include" are open-ended conjunctive verbs. Any one or more of the verb forms or tenses such as "comprises", "comprising", "has", "having", "includes", "including", etc. are open-ended. For example, any method of "comprises", "has", or "includes" one or more parts is not limited to having only those one or more parts, but also includes other unlisted parts.

[0068] "Pharmaceutically acceptable salts" are salts formed from acidic and basic groups of pentamidine analogs. Examples of such salts include acid addition salts or base addition salts such as hydrochloride, dihydrochloride, sulfate, citrate, hydrobromide, hydroiodide, nitrate, bisulfate, phosphate, metaphosphate, isonicotinate, acetate, lactate, salicylate, tartrate, pantothenate, ascorbate, succinate, maleate, fumarate, gluconate, saccharinate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate (pamoate)), etc., and salts of aluminum, calcium, lithium, magnesium, calcium, sodium, zinc, and diethanolamine. References to pentamidine analogs or pharmaceutically acceptable salts thereof should be understood to include pharmaceutically acceptable salts of the compounds disclosed herein. Examples of such pharmaceutically acceptable salts include, but are not limited to, isethionate, gluconate, and mesylate.

[0069] As used herein, the term "hydrogen" means a hydrogen atom (-H) and deuterium (heavy hydrogen, a non-radioactive isotope of hydrogen, D or 2 H). It should be understood that the present invention contemplates deuterated compound versions of all molecules of the present disclosure that can be synthesized by converting hydrogen atoms to 2 H wherever hydrogen atoms are present.

[0070] Pentamidine analogs In one aspect, formula (A):

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0071] In some variants of formula (A), [Chemical formula] , m, n, p, Z 1 , Z 2 , Y 1 -Y 10 , R 3 , R 4 , R 5 , R 6 , and R 7 are defined as above, and R 1 and R 2 are each independently hydrogen or halo, or R 1 is R 2 together with R 1 is R 2 together with R 1 and Z 2 both being O, one of Y 1 -Y 5 being N, one of Y 8 -Y 10 being N and the remaining Y 1 -Y 10 each being CH, and Y 1 -Y 5 together with the amidine substituent forms an amidine-substituted pyridine ring formed by Y 8 -Y 10 .

[0072] In some embodiments, at least one of Y 1 -Y 10 is N. In some embodiments, the ring formed by Y 1 -Y 5 is formed by Y6 -Y 10 is different from the ring formed by. In some embodiments, Y 1 -Y 10 The two amidine-substituted rings of are different so that the compound is not symmetric.

[0073] In some embodiments,

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0074] In some embodiments,

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0075] In some embodiments, [Chemical formula] The part [Chemical formula] , [Chemical formula] , [Chemical formula] , [Chemical formula] , and [Chemical formula] is selected from the group consisting of, [Chemical formula] The part [Chemical formula] , [Chemical formula] , [Chemical formula] , [Chemical formula] , and [Chemical formula] is selected from the group consisting of. In some embodiments, [Chemistry] is [Chemistry] , [Chemistry] , [Chemistry] , and [Chemistry] selected from the group consisting of. In some embodiments, [Chemistry] is [Chemistry] , [Chemistry] , [Chemistry] , and [Chemistry] selected from the group consisting of. In some embodiments, Y 1 -Y 10 The two amidine-substituted rings are different.

[0076] In some embodiments, [Chemistry] The moiety is [Chemistry] ,

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0077] In some embodiments,

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0078] In the description herein, all descriptions, modifications, embodiments or aspects of a part can be combined with all descriptions, modifications, embodiments or aspects of other parts as if all combinations of the descriptions were specifically and individually recited. For example, all descriptions, modifications, embodiments or aspects provided herein regarding R of formula (A) or (I)-(VIII) can be combined with all descriptions, modifications, embodiments or aspects of Y, X, n, m, and / or p as if all combinations were specifically and individually listed. It is understood that all descriptions, modifications, embodiments or aspects of formula (A) or (I)-(VIII) are equally applicable to and equally described by other formulas detailed herein as if the description, modification, embodiment, or aspect were specifically and individually listed separately for all formulas. For example, it is also understood that all descriptions, modifications, embodiments or aspects of formula (A) are equally applicable to and equally described by any applicable formula detailed herein such as formula (A)-(VIII) as if the description, modification, embodiment, or aspect were specifically and individually listed separately for all formulas.

[0079] In some embodiments, formula (I):

Chemical formula

Chemical formula

[0080] In some embodiments, at least one of Y 1 -Y 10 is N. In some embodiments, the ring formed by Y 1 -Y 5 is different from the ring formed by Y 6 -Y 10 . In some embodiments, the two amidine-substituted rings of Y 1 -Y 10 are different such that the compound is not symmetric.

[0081] In one embodiment, m is 1 and n is 1. In another embodiment, m is 1 and n is 0. In another embodiment, m is 0 and n is 1. In another embodiment, m is 1 and n is 2. In another embodiment, m is 2 and n is 1. In one embodiment, m is 2 and n is 2. In another embodiment, m is 0 and n is 0.

[0082] In one embodiment, Z 1 or Z 2 is independently selected from the group consisting of O, N, and S, each of which may be substituted. In one embodiment, Z 1 or Z 2 is independently O and may be substituted. In another embodiment, Z 1 or Z 2 is independently S and may be substituted. In yet another embodiment, Z 1 or Z 2 is independently NR 3 where R 3 is hydrogen. In one embodiment, Z 1 or Z 2 is independently NR 3 wherein R 3is alkyl, cycloalkyl, aryl, or heteroaryl. In another embodiment, Z 1 or Z 2 is, independently, NR 3 or CR 5 R 6 In another embodiment, Z 1 is NR 3 wherein R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, and Z 2 is CR 5 R 6 wherein R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R 5 together with R 6 forms a saturated or partially unsaturated 3- to 9-membered ring.

[0083] In one embodiment, the amidine is bonded independently by Y 3 and Y 8 In another embodiment, the amidine is bonded independently by Y 3 and Y 7 In yet another embodiment, the amidine is bonded independently by Y 2 and Y 7 In yet another embodiment, the amidine is bonded independently by Y 2 and Y 7

[0084] In one embodiment, Y 1、2、4、5、6、8 is CR 7 (e.g., -CH), Y 2 is N, and Y 3 and Y 7 are bonded to the amidine. In another embodiment, Y 1、4、5、6、及び7 is -CH, Y 2 is N, and Y 3 and Y 8 are CR 7 wherein R 7 is the amidine. In another embodiment, Y 1、4、5、6、及び8 is -CH, Y​3 is N, Y 2 and Y 7 is CR 7 wherein R 7 is amidine. In another embodiment, Y 1、4、5、6、及び8 is -CH, Y 3 is N, Y 2 and Y 7 is CR 7 wherein R 7 is amidine, m is 1, and n is 0. In another embodiment, Y 1、4、5、及び6 is -CH, Y 3 and Y 8 is N, Y 2 and Y 7 is CR 7 wherein R 7 is amidine, m is 1, and n is 0.

[0085] In one embodiment, R 1 and R 2 are independently hydrogen. In another embodiment, R 1 is R 2 which together with R forms a saturated, unsaturated, or partially unsaturated 3- to 9-membered cyclic group (e.g.,

Chemical formula

[0086] In one embodiment, R 3 is hydrogen. In another embodiment, R 3 is alkyl. For example, R 3 can be methyl or ethyl. In another embodiment, R 3is cycloalkyl. In another embodiment, R 3 is aryl. In yet another embodiment, R 3 is heteroaryl.

[0087] In one embodiment, R 4 is hydrogen. In another embodiment, R 4 is halo. In yet another embodiment, R 4 is cycloalkyl. In yet another embodiment, R 4 is aryl. In yet another embodiment, R 4 is heteroaryl. In one particular embodiment, R 4 is phenyl. In one embodiment, R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, amino, or, R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring. For example, R 5 and R 6 can be hydrogen. In one embodiment, R 7 is independently hydrogen or halo.

[0088] In some embodiments, the present invention relates to a compound having the formula (II):

Chemical formula

Chemical formula

[0089] In some embodiments, at least one X is N. In some embodiments, the two amidine-substituted rings are different such that the compound is not symmetric.

[0090] In one embodiment, m is 1 and n is 1. In another embodiment, m is 1 and n is 0. In another embodiment, m is 0 and n is 1. In another embodiment, m is 1 and n is 2. In another embodiment, m is 2 and n is 1. In one embodiment, m is 2 and n is 2. In another embodiment, m is 0 and n is 0.

[0091] In one embodiment, both X's are N. In another embodiment, one X is N or the other X is CR 7 is.

[0092] In one embodiment, Z 1 or Z 2 is independently selected from the group consisting of O, N, and S, each of which may be substituted. In one embodiment, Z 1 or Z 2 is independently O and may be substituted. In another embodiment, Z 1 or Z 2 is independently S and may be substituted. In yet another embodiment, Z 1 or Z 2 is independently NR 3 and R 3 is hydrogen. In one embodiment, Z 1 or Z 2 is independently NR 3 wherein R 3 is alkyl, cycloalkyl, aryl, or heteroaryl. In another embodiment, Z 1 or Z 2 is independently NR 3 or CR 5 R 6 is. In another embodiment, Z 1 is NR 3and wherein R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, and Z 2 is CR 5 R 6 and wherein R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring.

[0093] In one embodiment, R 1 and R 2 are independently hydrogen. In another embodiment, R 1 and R 2 together form a saturated, unsaturated, or partially unsaturated 3- to 9-membered cyclic group (e.g.,

Chemical formula

[0094] In some embodiments, the present invention relates to a compound having the formula (III):

Chemical formula

Chemical formula

[0095] In some embodiments, at least one X is N. In some embodiments, the two amidine-substituted rings are different such that the compound is not symmetric.

[0096] In one embodiment, m is 1 and n is 1. In another embodiment, m is 1 and n is 0. In another embodiment, m is 0 and n is 1. In another embodiment, m is 1 and n is 2. In another embodiment, m is 2 and n is 1. In one embodiment, m is 2 and n is 2. In another embodiment, m is 0 and n is 0.

[0097] In one embodiment, both X's are N. In another embodiment, one X is N or the other X is CR 7 is.

[0098] In one embodiment, Z 1 or Z 2 is independently selected from the group consisting of O, N, and S, each of which may be substituted. In one embodiment, Z 1 or Z 2 is independently O, which may be substituted. In another embodiment, Z 1 or Z 2 is independently S, which may be substituted. In yet another embodiment, Z 1 or Z 2 is independently NR 3 and R 3 is hydrogen. In one embodiment, Z 1 or Z 2 is independently NR 3 where R 3 is alkyl, cycloalkyl, aryl, or heteroaryl. In another embodiment, Z 1 or Z 2 is independently NR 3 or CR 5 R 6 In another embodiment, Z 1 is NR 3 where R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, and Z 2 is CR 5 R 6 where R 5 or R 6is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R 5 is R 6 together with forms a saturated or partially unsaturated 3- to 9-membered ring.

[0099] In one embodiment, R 1 and R 2 are independently hydrogen. In another embodiment, R 1 is R 2 together with forms a saturated, unsaturated, or partially unsaturated 3- to 9-membered cyclic group (e.g.,

Chemical formula

[0100] In some embodiments, the present invention relates to a compound having the formula (IV):

Chemical formula

Chemical formula

[0101] In some embodiments, the two amidine-substituted rings are different such that the compound is not symmetric.

[0102] In one embodiment, m is 1 and n is 1. In another embodiment, m is 1 and n is 0. In another embodiment, m is 0 and n is 1. In another embodiment, m is 1 and n is 2. In another embodiment, m is 2 and n is 1. In one embodiment, m is 2 and n is 2. In another embodiment, m is 0 and n is 0.

[0103] In one embodiment, both X's are N. In another embodiment, one X is N or the other X is CR 7 is.

[0104] In one embodiment, Z 1 or Z 2 is independently selected from the group consisting of O, N, and S, each of which may be substituted. In one embodiment, Z 1 or Z 2 is independently O and may be substituted. In another embodiment, Z 1 or Z 2 is independently S and may be substituted. In yet another embodiment, Z 1 or Z 2 is independently NR 3 and R 3 is hydrogen. In one embodiment, Z 1 or Z 2 is independently NR 3 where R 3 is alkyl, cycloalkyl, aryl, or heteroaryl. In another embodiment, Z 1 or Z 2 is independently NR 3 or CR 5 R 6 is. In another embodiment, Z 1 is NR 3 where R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl and Z 2 is CR 5 R 6 where R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R 5 together with R 6 forms a saturated or partially unsaturated 3- to 9-membered ring.

[0105] In one embodiment, R 1 and R 2 are independently hydrogen. In another embodiment, R1 is R 2 together with R forms a saturated, unsaturated, or partially unsaturated 3- to 9-membered cyclic group (e.g.,

Chemical formula

[0106] In some embodiments, the present invention relates to a compound having the formula (V):

Chemical formula

[0107] In some embodiments, at least one of Y 1 -Y 10 is N. In some embodiments, the ring formed by Y 1 -Y 5 is different from the ring formed by Y 6 -Y 10 . In some embodiments, the two amidine-substituted rings of Y 1 -Y 10 are different so that the compound is not symmetric.

[0108] In one embodiment, Z 1 or Z 2 is independently selected from the group consisting of O, N, and S and may be substituted. In one embodiment, Z 1 or Z 2 is independently O and may be substituted. In another embodiment, Z 1 or Z 2 is independently S and may be substituted. In yet another embodiment, Z 1 or Z 2 is independently NR 3 and R 3 is hydrogen. In one embodiment, Z 1 or Z2 is independently NR 3 wherein R 3 is alkyl, cycloalkyl, aryl, or heteroaryl. In another embodiment, Z 1 or Z 2 is independently NR 3 or CR 5 R 6 wherein in another embodiment, Z 1 is NR 3 wherein R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, and Z 2 is CR 5 R 6 wherein R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring. In one embodiment, Z 1 and Z 2 are CR 5 R 6 wherein R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, or amino, or R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring.

[0109] In one embodiment, the amidine is independently bonded by Y 3 and Y 8 In another embodiment, the amidine is independently bonded by Y 3 and Y 7 In yet another embodiment, the amidine is independently bonded by Y 2 and Y 7 In yet another embodiment, the amidine is independently bonded by Y 2 and Y 7

[0110] In one embodiment, Y​1、2、4、5、6、8 is CR 7 (for example, -CH), and Y 2 is N, and Y 3 and Y 7 is bonded to amidine. In another embodiment, Y 1、4、5、6、及び7 is -CH, and Y 2 is N, and Y 3 and Y 8 is CR 7 wherein R 7 is amidine. In another embodiment, Y 1、4、5、6、及び8 is -CH, and Y 3 is N, and Y 2 and Y 7 is CR 7 wherein R 7 is amidine. In another embodiment, Y 1、4、5、6、及び8 is -CH, and Y 3 is N, and Y 2 and Y 7 is CR 7 wherein R 7 is amidine. In another embodiment, Y 1、4、5、及び6 is -CH, and Y 3 and Y 8 is N, and Y 2 and Y 7 is CR 7 wherein R 7 is amidine.

[0111] In one embodiment, R 3 is hydrogen. In another embodiment, R 3 is alkyl. For example, R 3 can be methyl or ethyl. In another embodiment, R 3 is cycloalkyl. In another embodiment, R 3 is aryl. In yet another embodiment, R 3 is heteroaryl. In one embodiment, R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, amino, or R 5 where R 6Together, they form a saturated or partially unsaturated 3- to 9-membered ring. For example, R 5 and R 6 can be hydrogen. In one embodiment, R 7 is independently hydrogen or halo. In one embodiment, R 8 is independently hydrogen, halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino. In one embodiment, R 8 is hydrogen.

[0112] In some embodiments, the present invention relates to a compound having the formula (VI):

Chemical formula

[0113] In some embodiments, at least one X is N. In some embodiments, the two amidine-substituted rings are different such that the compound is not symmetric.

[0114] In one embodiment, Z 1 or Z 2 is independently selected from the group consisting of O, N, and S and may be substituted. In one embodiment, Z 1 or Z 2 is independently O and may be substituted. In another embodiment, Z 1 or Z 2 is independently S and may be substituted. In yet another embodiment, Z 1or Z 2 is independently NR 3 and R 3 is hydrogen. In one embodiment, Z 1 or Z 2 is independently NR 3 wherein R 3 is alkyl, cycloalkyl, aryl, or heteroaryl. In another embodiment, Z 1 or Z 2 is independently NR 3 or CR 5 R 6 In another embodiment, Z 1 is NR 3 wherein R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, and Z 2 is CR 5 R 6 wherein R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring. In one embodiment, Z 1 and Z 2 are CR 5 R 6 wherein R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, or amino, or R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring.

[0115] In one embodiment, R 3 is hydrogen. In another embodiment, R 3 is alkyl, for example, R 3 can also be methyl or ethyl. In another embodiment, R 3 is cycloalkyl. In another embodiment, R 3 is aryl. In yet another embodiment, R 3is heteroaryl. In one embodiment, R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, amino, or R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring. For example, R 5 and R 6 can be hydrogen. In one embodiment, R 7 is independently hydrogen or halo. In one embodiment, R 8 is independently hydrogen, halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino. In one embodiment, R 8 is hydrogen.

[0116] In some embodiments, the present invention relates to a compound having the formula (VII):

Chemical formula

[0117] In some embodiments, at least one X is N. In some embodiments, the two amidine-substituted rings are different such that the compound is not symmetric.

[0118] In one embodiment, Z 1 or Z 2 is independently selected from the group consisting of O, N, and S and may be substituted. In one embodiment, Z1 or Z 2 is independently O and may be substituted. In another embodiment, Z 1 or Z 2 is independently S and may be substituted. In yet another embodiment, Z 1 or Z 2 is independently NR 3 and R 3 is hydrogen. In one embodiment, Z 1 or Z 2 is independently NR 3 wherein R 3 is alkyl, cycloalkyl, aryl, or heteroaryl. In another embodiment, Z 1 or Z 2 is independently NR 3 or CR 5 R 6 In another embodiment, Z 1 is NR 3 wherein R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, and Z 2 is CR 5 R 6 wherein R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring. In one embodiment, R 3 is hydrogen. In another embodiment, R 3 is alkyl, for example, R 3 can also be methyl or ethyl. In another embodiment, R 3 is cycloalkyl. In another embodiment, R 3 is aryl. In yet another embodiment, R 3 is heteroaryl. In one embodiment, R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, amino, or R 5 and R 6Together they form a saturated or partially unsaturated 3- to 9-membered ring. For example, R 5 and R 6 can be hydrogen. In one embodiment, R 7 is independently hydrogen or halo. In one embodiment, R 8 is independently hydrogen, halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino. In one embodiment, R 8 is hydrogen.

[0119] In some embodiments, the present invention relates to a compound having the formula (VIII):

Chemical formula

[0120] In some embodiments, at least one X is N. In some embodiments, the two amidine-substituted rings are different such that the compound is not symmetric.

[0121] In one embodiment, Z 1 or Z 2 is independently selected from the group consisting of O, N, and S and may be substituted. In one embodiment, Z 1 or Z 2 is independently O and may be substituted. In another embodiment, Z 1 or Z 2is independently S and may be substituted. In yet another embodiment, Z 1 or Z 2 is independently NR 3 and R 3 is hydrogen. In one embodiment, Z 1 or Z 2 is independently NR 3 wherein R 3 is alkyl, cycloalkyl, aryl, or heteroaryl. In another embodiment, Z 1 or Z 2 is independently NR 3 or CR 5 R 6 In another embodiment, Z 1 is NR 3 wherein R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, and Z 2 is CR 5 R 6 wherein R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring. In one embodiment, R 3 is hydrogen. In another embodiment, R 3 is alkyl, for example, R 3 can also be methyl or ethyl. In another embodiment, R 3 is cycloalkyl. In another embodiment, R 3 is aryl. In yet another embodiment, R 3 is heteroaryl. In one embodiment, R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, amino, or R 5 and R 6 together form a saturated or partially unsaturated 3- to 9-membered ring. For example, R 5 and R 6 can be hydrogen. In one embodiment, R7 is independently hydrogen or halo. In one embodiment, R 8 is independently hydrogen, halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino. In one embodiment, R 8 is hydrogen.

[0122] Exemplary compounds include compounds of the following structures, or pharmaceutically acceptable salts thereof:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0123] method Compounds of any formula provided herein, or salts and pharmaceutically acceptable carriers or excipients thereof, such as pharmaceutical compositions, the compounds and compositions detailed herein can be used in the methods of administration and treatment provided herein. It is understood that any of the methods or pharmaceutical compositions detailed herein can use a compound of any formula detailed herein, or a variant thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, for example, any of the methods or pharmaceutical compositions detailed herein, such as a method involving administering to a subject a compound of formula (A) or a pharmaceutically acceptable salt thereof, uses a compound of formula (A), or a pharmaceutically acceptable salt thereof. In one embodiment, any of the methods or pharmaceutical compositions detailed herein uses a compound of formula (I), or a pharmaceutically acceptable salt thereof. In one embodiment, any of the methods or pharmaceutical compositions detailed herein uses a compound of formula (II), or a pharmaceutically acceptable salt thereof. In one embodiment, any of the methods or pharmaceutical compositions detailed herein uses a compound of formula (III), or a pharmaceutically acceptable salt thereof. In one embodiment, any of the methods or pharmaceutical compositions detailed herein uses a compound of formula (IV), or a pharmaceutically acceptable salt thereof. In one embodiment, any of the methods or pharmaceutical compositions detailed herein uses a compound of formula (V), or a pharmaceutically acceptable salt thereof. In one embodiment, any of the methods or pharmaceutical compositions detailed herein uses a compound of formula (VI), or a pharmaceutically acceptable salt thereof. In one embodiment, any of the methods or pharmaceutical compositions detailed herein uses a compound of formula (VII), or a pharmaceutically acceptable salt thereof. In one embodiment, any of the methods or pharmaceutical compositions detailed herein uses a compound of formula (VIII), or a pharmaceutically acceptable salt thereof. In one embodiment, any of the methods or pharmaceutical compositions detailed herein uses a compound of Table I, or a pharmaceutically acceptable salt thereof. Thus, in one embodiment, any method or pharmaceutical composition detailed herein is of the formula

Chemical formula

[0124] There is provided a method of treating cancer comprising administering the compound described herein to a subject in need thereof. In some embodiments, the method comprises treating a solid tumor. In some embodiments, the method comprises treating a cancer selected from the group consisting of liver cancer, cholangiocarcinoma, colon cancer, hepatobiliary duct cancer, and renal cancer. In some embodiments, the subject is a human.

[0125] The compounds and compositions described herein can be administered to a subject in need of treatment for a cell proliferation disorder such as cancer, particularly cancer selected from the group consisting of liver cancer, cholangiocarcinoma, osteosarcoma, melanoma, breast cancer, renal cancer, prostate cancer, gastric cancer, colorectal cancer, thyroid cancer, head and neck cancer, ovarian cancer, pancreatic cancer, neuroblastoma, lung cancer, uterine cancer, leukemia, or lymphoma. The subject is typically a mammal diagnosed as in need of treatment for one or more of such proliferative disorders, and in many cases, the subject is a human. The method comprises administering an effective amount of at least one compound of the invention. Optionally, the compound can be administered in combination with one or more additional therapeutic agents, particularly therapeutic agents known to be useful for treating the cancer or proliferative disorder afflicting a particular subject. It will be understood by those skilled in the art that colorectal cancer and colon cancer are used interchangeably herein, and renal cancer and kidney cancer are used interchangeably herein.

[0126] The compounds of the present disclosure or their pharmaceutically acceptable salts are generally administered in a therapeutically effective amount. The term "therapeutically effective amount" can refer to that amount (or dosage) of the compound or other treatment that is necessary and sufficient to prevent, mitigate, ameliorate, treat, or eliminate a medical condition or the risk thereof when administered to a subject in need of the compound or other treatment. The amount of the compound actually administered to a subject can be determined by a physician or caregiver according to relevant circumstances including the medical condition being treated, the selected route of administration, the compound being administered and its relative activity, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like. Accordingly, a therapeutically effective amount can vary, for example, depending on the medical condition of the subject, the weight and age of the subject, the severity of the medical condition, the method of administration, and the like.

[0127] The compounds of the present disclosure can be administered by any of the acceptable modes of administration of agents having similar utility, such as, for example, oral, dermal, topical, intradermal, intrathecal, intravenous, subcutaneous, intramuscular, intra-articular, intraspinal or intramedullary, nasal, epidural, rectal, vaginal, or intradermal / transmucosal routes. The appropriate route varies depending on the nature and severity of the medical condition being treated. Oral administration can be the primary route of administration of the compounds of the present disclosure as it generally exhibits increased oral bioavailability, enhanced organ targeting in combination with reduced in vivo toxicity. However, intravenous (IV) administration can also be a route of administration of the compounds of the present disclosure. Intramuscular (IM) administration can be a route of administration of the compounds of the present disclosure. Subcutaneous, sublingual, or transdermal administration can also be contemplated as routes of administration of the compounds of the present disclosure. Sublingual administration can be carried out using an appropriate formulation of the compound. Inhalation administration can also be used as a route of administration using an appropriate formulation for compounds and cancer types (e.g., lung cancer) that can benefit from this route.

[0128] In certain examples, the pharmaceutical compositions provided herein can be orally administered to a human patient at a dose of from about 0.1 mg / kg to about 300 mg / kg, or even 500 mg / kg. In another embodiment, the pharmaceutical compositions provided herein can be orally administered daily to a human patient at a dose of from about 1 mg / kg to about 300 mg / kg. In another specific example, the pharmaceutical compositions provided herein can be orally administered to a human patient at a dose of from about 1 mg / kg to about 100 mg / kg.

[0129] The subject may be at risk of having cancer. The subject can be a mammal. The subject may be a human patient suffering from cancer. Examples of cancers include, but are not limited to, adrenal cancer, anal cancer, bile duct cancer, bladder cancer, blood cancer, bone cancer, brain tumor, breast cancer, cardiovascular cancer, cervical cancer, colon cancer, digestive system cancer, endocrine system cancer, endometrial cancer, esophageal cancer, eye cancer, gallbladder cancer, gastrointestinal tumor, kidney cancer, laryngeal cancer, leukemia, liver cancer, lung cancer, bile duct cancer, lymphoma, mesothelioma, muscle system cancer, myelodysplastic syndrome, multiple myeloma, nasal cancer, nasopharyngeal cancer, nervous system cancer, lymphatic system cancer, oral cancer, oropharyngeal cancer, ovarian cancer, pancreatic cancer, penile cancer, pituitary tumor, prostate cancer, genital system cancer, respiratory system cancer, sarcoma, salivary gland cancer, skeletal system cancer, skin cancer, small intestine cancer, stomach cancer, testicular cancer, thymus cancer, thyroid cancer, bladder cancer, or vaginal cancer. In one embodiment, the subject is suffering from liver cancer. In another embodiment, the subject is suffering from bile duct cancer. In another embodiment, the subject is suffering from hepatobiliary duct cancer. In yet another embodiment, the subject is suffering from kidney cancer. In yet another embodiment, the solid tumor is suffering from colon cancer. In yet another embodiment, the subject is suffering from lung cancer (e.g., small cell lung cancer or non-small cell lung cancer). In yet another embodiment, the subject is suffering from breast cancer. In yet another embodiment, the subject is suffering from ovarian cancer.

[0130] Examples of cancers include cancers that cause solid tumors and cancers that do not cause solid tumors. Further, any of the cancers referred to herein can be a primary cancer (e.g., a cancer named after the part of the body where it first began to grow) or a secondary or metastatic cancer (e.g., a cancer that originated from another part of the body).

[0131] In some embodiments, provided herein is a method of inhibiting cancer cell growth in an individual, comprising administering to the individual a compound provided herein. In some embodiments, at least about 10% (including any of, for example, at least about 20%, about 30%, about 40%, about 60%, about 70%, about 80%, about 90%, or about 100%) of cell growth is inhibited. In some embodiments, the growth of solid tumors is inhibited. In some embodiments, the growth of liver cancer cells is inhibited. In some embodiments, the growth of colon cancer cells is inhibited. In some embodiments, the growth of kidney cancer cells is inhibited. In some embodiments, the growth of cholangiocarcinoma cells is inhibited.

[0132] Also provided herein is a method of inhibiting tumor metastasis in an individual, comprising administering to the individual a compound provided herein. In some embodiments, at least about 10% (including any of, for example, at least about 20%, about 30%, about 40%, about 60%, about 70%, about 80%, about 90%, or about 100%) of the translocation is inhibited. In some embodiments, the metastasis of liver cancer is inhibited. In some embodiments, the metastasis of colon cancer cells is inhibited. In some embodiments, the metastasis of kidney cancer cells is inhibited. In some embodiments, the metastasis of cholangiocarcinoma is inhibited. In any of the above embodiments, metastasis to lymph nodes, lungs, bones, or the brain is inhibited. In any of the above embodiments, tumor metastasis can be inhibited for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks after treatment.

[0133] In some embodiments, the method comprises reducing the tumor size and / or tumor burden of the individual. In some embodiments, the tumor size is reduced by at least about 10% (including any of, for example, at least about 20%, about 30%, about 40%, about 60%, about 70%, about 80%, about 90%, or about 100%). In some embodiments, the tumor is liver cancer. In some embodiments, the tumor is kidney cancer. In some embodiments, the tumor is colon cancer. In some embodiments, the tumor is cholangiocarcinoma.

[0134] In some embodiments, the method includes prolonging the progression-free survival of an individual. In some embodiments, the method prolongs the progression-free disease period by any of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks. In some embodiments, the individual has a solid tumor. In some embodiments, the individual has liver cancer. In some embodiments, the individual has kidney cancer. In some embodiments, the individual has colon cancer. In some embodiments, the individual has cholangiocarcinoma.

[0135] In some embodiments, the method includes alleviating one or more symptoms of an individual having cancer. In some embodiments, the individual has a solid tumor. In some embodiments, the individual has liver cancer. In some embodiments, the individual has kidney cancer. In some embodiments, the individual has colon cancer. In some embodiments, the individual has cholangiocarcinoma.

[0136] In some embodiments, the method includes improving the quality of life of an individual having cancer. In some embodiments, the individual has a solid tumor. In some embodiments, the individual has liver cancer. In some embodiments, the individual has kidney cancer. In some embodiments, the individual has colon cancer. In some embodiments, the individual has cholangiocarcinoma.

[0137] In some embodiments, the method results in an objective response (such as a partial response or a complete response) in a patient having cancer. In some embodiments, the individual has a solid tumor. In some embodiments, the individual has liver cancer. In some embodiments, the individual has kidney cancer. In some embodiments, the individual has colon cancer. In some embodiments, the individual has cholangiocarcinoma.

[0138] In some embodiments, the compounds of the invention are not metabolized by cytochrome P-450 and result in a reduction in toxicity, particularly hepatotoxicity, compared to existing therapies. Thus, in some embodiments, provided herein is a method of treating cancer in an individual with reduced liver function. In some embodiments, the individual has a Child-Pugh score of class B or class C.

[0139] In some embodiments, the method results in a decrease in one or more markers of liver injury or tumor burden in an individual having liver cancer. In some embodiments, the method results in a decrease in the level of one or more of alanine aminotransferase (ALT), aspartate aminotransferase (AST), or alkaline phosphatase (ALP). In some embodiments, the level of the liver injury marker decreases by at least about 5% (e.g., about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90%).

[0140] The compounds of the present disclosure or pharmaceutically acceptable salts thereof can be administered to a subject (e.g., a human patient) suffering from cancer, for example, orally, intravenously or subcutaneously, at a dose of, for example, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, about 100 mg / kg, about 110 mg / kg, about 120 mg / kg, about 130 mg / kg, about 140 mg / kg, about 150 mg / kg, about 160 mg / kg, about 170 mg / kg, about 180 mg / kg, about 190 mg / kg, about 200 mg / kg, about 210 mg / kg, about 220 mg / kg, about 230 mg / kg, about 240 mg / kg, about 250 mg / kg, about 260 mg / kg, about 270 mg / kg, about 280 mg / kg, about 290 mg / kg, about 300 mg / kg, about 350 mg / kg, about 400 mg / kg, about 450 mg / kg, about 500 mg / kg, or about 600 mg / kg.

[0141] In one embodiment, the compound of the present disclosure or a pharmaceutically acceptable salt thereof can be orally administered, for example, at a dose of about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, about 100 mg / kg, about 110 mg / kg, about 120 mg / kg, about 130 mg / kg, about 140 mg / kg, about 150 mg / kg, about 160 mg / kg, about 170 mg / kg, about 180 mg / kg, about 190 mg / kg, about 200 mg / kg, about 210 mg / kg, about 220 mg / kg, about 230 mg / kg, about 240 mg / kg, about 250 mg / kg, about 260 mg / kg, about 270 mg / kg, about 280 mg / kg, about 290 mg / kg, about 300 mg / kg, about 350 mg / kg, about 400 mg / kg, about 450 mg / kg, about 500 mg / kg, or about 600 mg / kg.

[0142] In one embodiment, the compound of the present disclosure or a pharmaceutically acceptable salt thereof can be administered intravenously at a dose of, for example, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 35 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, about 100 mg / kg, about 110 mg / kg, about 120 mg / kg, about 130 mg / kg, about 140 mg / kg, about 150 mg / kg, about 160 mg / kg, about 170 mg / kg, about 180 mg / kg, about 190 mg / kg, about 200 mg / kg, about 210 mg / kg, about 220 mg / kg, about 230 mg / kg, about 240 mg / kg, about 250 mg / kg, about 260 mg / kg, about 270 mg / kg, about 280 mg / kg, about 290 mg / kg, or about 300 mg / kg.

[0143] In one embodiment, the compound of the present disclosure or a pharmaceutically acceptable salt thereof can be administered subcutaneously at a dose of, for example, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, about 100 mg / kg, about 110 mg / kg, about 120 mg / kg, about 130 mg / kg, about 140 mg / kg, about 150 mg / kg, about 160 mg / kg, about 170 mg / kg, about 180 mg / kg, about 190 mg / kg, about 200 mg / kg, about 210 mg / kg, about 220 mg / kg, about 230 mg / kg, about 240 mg / kg, about 250 mg / kg, about 260 mg / kg, about 270 mg / kg, about 280 mg / kg, about 290 mg / kg, or about 300 mg / kg.

[0144] In one embodiment, the compound of the present disclosure or a pharmaceutically acceptable salt thereof can be administered orally to a subject suffering from liver cancer at a dose of, for example, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, about 100 mg / kg, about 110 mg / kg, about 120 mg / kg, about 130 mg / kg, about 140 mg / kg, about 150 mg / kg, about 160 mg / kg, about 170 mg / kg, about 180 mg / kg, about 190 mg / kg, or about 200 mg / kg.

[0145] Administration can be three times a day, twice a day, once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every ten days, or once every two weeks. Administration can also include a dosing holiday (s) of about one day to about seven days between administrations.

[0146] Combination therapy The disclosure provided herein describes a method of treating cancer in a subject by administering at least one compound of the disclosure to the subject. The methods disclosed herein can further include administering a combination of a compound of formula (I)-(VIII) or a pharmaceutically acceptable salt thereof and at least one additional anti-cancer agent, wherein the combined composition can be administered as a co-formulation or separately.

[0147] In some particular embodiments, two or more compounds of the disclosure can be administered to a subject at once. In some embodiments, the combination of two compounds of the disclosure can act synergistically or additively, and either compound can be used in a lesser amount than when administered alone.

[0148] In some embodiments, the compounds and / or pharmaceutical compositions thereof disclosed herein are administered concurrently with the administration of another therapeutic agent. For example, the compounds and / or pharmaceutical compositions thereof disclosed herein can be administered together with another therapeutic agent. In other embodiments, the compounds and / or pharmaceutical compositions thereof disclosed herein are administered before or after the administration of another therapeutic agent.

[0149] In combination therapy, the additional therapeutic agents used in the combination therapy can be anti-cancer agents such as etoposide, cisplatin, oxaliplatin, gemcitabine, irinotecan, anthracycline, and taxol.

[0150] As used herein, the term "therapeutically effective amount" means an amount of a pentamidine analog such as formula (I)-(VIII) or a pharmaceutically acceptable salt thereof.

[0151] The dosage of the compound administered in accordance with the present invention is determined taking into account the specific circumstances surrounding the case, including, for example, the compound administered, the route of administration, the patient's medical condition, the stage of the cancer, and the physical properties of the anticancer agent used in combination therapy.

[0152] For administration to a subject, the pentamidine analog, or a pharmaceutically acceptable salt thereof, is generally formulated in a pharmaceutical composition and a pharmaceutically acceptable carrier. The therapeutic composition is generally sterile and sufficiently stable under the conditions of manufacture and storage.

[0153] Many types of anticancer approaches can be used in combination with a pentamidine analog, or a pharmaceutically acceptable salt thereof, including, for example, chemotherapeutic agents, biological agents, radiation, and surgical therapies. The methods of the present invention can use these approaches to treat the same types of cancer as those used in the art. Further, these approaches can be carried out according to parameters (e.g., regimens and dosages) similar to those known in the art for their use.

[0154] For example, several different types of chemotherapeutic agents, including antimetabolites, antibiotics, alkylating agents, plant alkaloids, hormonal agents, anticoagulants, antithrombotic agents, and other natural products, can be used in combination with the pentamidine analogs disclosed herein.

[0155] Many approaches for administering anti-cancer agents are known in the art and can be readily adapted for use in the present invention. The preferred route of administration is oral administration for combination therapy. In the case of systemic administration, the drug can be administered, for example, by intravenous injection or infusion (continuous or bolus). The appropriate schedule and dosage of such administration can be readily determined by those skilled in the art based on, for example, preclinical studies in animals and clinical studies in humans (e.g., Phase I studies). Many regimens used for administering chemotherapeutic agents include, for example, intravenous administration of one or more agents followed by repeating this treatment after a period (e.g., 1 to 4 weeks) during which the patient recovers from the harmful side effects of the treatment. It may be desirable to use both agents in each administration or to include only one agent in part (or all) of the treatment.

[0156] Pharmaceutical formulation The compounds of the present disclosure can be administered by any of the acceptable modes of administration of agents having similar utility, such as, for example, oral, dermal, topical, intradermal, intrathecal, intravenous, subcutaneous, intramuscular, intra-articular, intraspinal or intramedullary, nasal, epidural, or inhalable via intradermal / transmucosal routes.

[0157] In one specific example, the pharmaceutical composition can be administered orally to a patient. In another specific example, a pharmaceutical composition containing pentamidine or a pharmaceutically acceptable salt thereof can be administered intravenously to a patient (e.g., by injection or infusion). In another specific example, the pharmaceutical composition can be administered intramuscularly to a patient. In one specific example, the pharmaceutical composition can be administered nasally to a patient. The pharmaceutical composition (e.g., for oral administration or inhalation, injection, infusion, subcutaneous delivery, intramuscular delivery, intraperitoneal delivery, sublingual delivery, or other methods) can be in liquid form. The liquid pharmaceutical composition can include, for example, water, physiological saline, preferably physiological saline, Ringer's solution, isotonic sodium chloride, a fixed oil that can function as a solvent or suspension medium, polyethylene glycol, glycerin, propylene glycol, or other solvents; an antibacterial agent; an antioxidant; a chelating agent; a buffer for adjusting tonicity such as sodium chloride or dextrose; and one or more of the agents. The parenteral composition can be enclosed in an ampoule, a disposable syringe, or a multiple-dose vial made of glass or plastic. The use of physiological saline is preferred, and the injectable pharmaceutical composition is preferably sterile. The liquid pharmaceutical composition can be delivered orally.

[0158] A pharmaceutical composition containing a compound of formula (I) - (VIII) or a pharmaceutically acceptable salt thereof can be formulated for sustained release or controlled release (also referred to as extended release or slow release). Such compositions can be prepared using well-known techniques and can be administered, for example, orally, rectally, intradermally, or subcutaneously, or by implantation at the desired target site. The controlled release formulation can include a compound dispersed in a carrier matrix and / or contained within a reservoir surrounded by a rate-controlling membrane. The excipients used in such formulations can be biocompatible and can be biodegradable. Preferably, the formulation provides a relatively constant level of active ingredient release. Non-limiting examples of excipients include water, alcohol, glycerol, chitosan, alginate, chondroitin, vitamin E, mineral oil, and dimethyl sulfoxide (DMSO). The amount of the compound included in the controlled release formulation is determined by the implantation site, the rate of release and the expected duration, as well as the nature of the condition, disease or disorder to be treated or prevented.

[0159] A pharmaceutical composition comprising one or more pentamidine analogs or a pharmaceutically acceptable salt thereof may be effective over time. In some cases, the pharmaceutical composition may be effective for one day or more. In some cases, the effective period of the pharmaceutical composition may be over a long period. In some cases, the effectiveness of the pharmaceutical composition may exceed 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 1 month.

[0160] When preparing a pharmaceutical composition comprising one or more pentamidine analogs or a pharmaceutically acceptable salt thereof, the active ingredient can be diluted with an excipient. Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, PEG, polyvinylpyrrolidone, cellulose, water, sterile saline, syrup, and methylcellulose. The compositions of the present disclosure can be formulated to provide rapid, sustained, or delayed release of the active ingredient after administration to a patient by using procedures known in the art. In some cases, a pharmaceutical composition comprising pentamidine or a pharmaceutically acceptable salt thereof may include an excipient that provides long-term storage, increases the bulk of a formulation containing a potent active ingredient, promotes drug absorption, reduces viscosity, adds flavor, or enhances the solubility of the pharmaceutical composition.

[0161] In some cases, a pharmaceutical composition comprising a pentamidine analog or a pharmaceutically acceptable salt thereof may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers include any physiologically compatible solvent, dispersion medium, coating, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. Preferably, the carrier is suitable for oral administration. The active compound may be coated in a substance to protect the compound from the action of acids and other natural conditions that may inactivate the compound. The carrier may be suitable for parenteral (e.g., intravenous, intramuscular, subcutaneous, intrathecal) administration (e.g., by injection or infusion).

[0162] The present invention also contemplates formulating pharmaceutically acceptable salts of the compounds of formulas (I)-(VIII). Generally, pharmaceutical salts include, but are not limited to, salts and base addition salts (e.g., hydrochloride, dihydrochloride, sulfate, citrate, hydrobromide, hydroiodide, nitrate, bisulfate, phosphate, superphosphate, isonicotinate, acetate, lactate, salicylate, tartrate, pantothenate, ascorbate, succinate, maleate, fumarate, gluconate, saccharinate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate (pamoate)), and salts of aluminum, calcium, lithium, magnesium, calcium, sodium, zinc, and diethanolamine.

[0163] The present invention is further illustrated by the following examples, which should not be construed as further limitations. The contents of all references, patents, and published patent applications cited throughout this application, as well as the sequence diagrams and appendices provided herein, are hereby expressly incorporated by reference in their entirety.

Example

[0164] Analogs of pentamidine were designed and synthesized using the synthetic methods described further below (see Table 1).

Chemical formula

[0165] General Information 1 1H NMR spectra and 13 13C NMR spectra were recorded on a Varian 400 MHz or Bruker Avance III 500 MHz spectrometer. Unless otherwise noted, spectra were referenced to residual chloroform (δ 7.26, 1 1H), DMSO (δ 2.54, 1 1H) or methanol (δ 3.34, 1It is based on (H). Chemical shifts are reported in ppm (δ). Multiplicities are indicated by s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), sext (sextet), m (multiplet), and br (broad). Coupling constants J are reported in Hertz. Silica gel chromatography was performed using a Teledyne Isco ComniFlash® Rf+ instrument with a Hi-Purit silica flash cartridge (National Chromatography Inco) or a RediSep Rf Gold C18 cartridge (Teledyne Isco). Analytical HPLC was performed on a Waters ACQUITY UPLC equipped with a photodiode array detector using a Waters ACQUITY BEH Shield RPC18 (2.1×50 mm, 1.7 μm) column. Analytical LCMS was performed on a Waters ACQUITY UPLC equipped with a Waters 3100 mass detector. Chiral HPLC was performed on a Waters Alliance e2695 equipped with a photodiode array detector using a Daicel Chiralpak® AD-H, Chiralpak® IA, Chiralpak® IB, Chiralpak® IC, Chiralcel® OD-H, or Chiralcel® OJ-H column. Optical rotations were obtained with a Jasco P-2000 digital polarimeter, [□] D T The temperature (T), concentration (c = g / 100 mL), and solvent are reported. Unless otherwise specified, commercially available reagents and solvents were used as received.

Table 1

[0166] Example 1: Preparation of 5-(5-(4-carbamimidoylphenoxy)pentyloxy)picolylimidamide

Chem.

[0167] Project 1 To a stirred solution of 4-hydroxybenzonitrile (10 g, 0.08 mol, 1 equiv) in acetone (120 mL) was added 1,5-dibromopentane (95.72 g, 0.42 mol, 5 equiv) and potassium carbonate (23.21 g, 0.16 mol, 2 equiv). Next, the reaction mixture was stirred at 7 °C for 2 h. The reaction mixture was monitored by TLC-LC-MS. The reaction mixture was diluted with water (500 mL) and extracted with EtOAC (2 × 800 mL). The separated organic layer was dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure to give a crude product. The crude product was purified by a glass column to give 4-((5-bromopentyl)benzonitrile (15 g, 66.94%). Analysis data LCMS: 268 [M+1] +

[0168] Project 2 To a stirred solution of 4-((5-bromopentyl)benzonitrile (0.52 g, 4.33 mmol, 1 equiv) in acetone (5 mL) was added compound 5-hydroxypicolinonitrile (1.15 g, 4.33 mmol, 1 equiv) and potassium carbonate (1.19 g, 8.66 mmol, 2 equiv) at 70 °C for 2 h. The reaction mixture was monitored by TLC and LC-MS. The reaction mixture was diluted with water (80 ml) and extracted with EtOAC (400 ml). The separated organic layer was dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure to give a crude product. The crude product was purified by combiflash chromatography to give 5-((5-(cyanophenoxy)pentyl)oxy)picolinonitrile (0.6 g, 45.11%). Analysis data LCMS: 308 ([M+1]) +

[0169] Project 3 NH in toluene (5 mL) 4To a stirred suspension of Cl (0.27 g, 0.65 mmol, 8 eq) at 0 °C, trimethylaluminum (0.27 g, 5.21 mmol, 8 eq) was added. The reaction mixture was stirred at 0 °C for 10 minutes and then at room temperature for 15 minutes. To this solution, 5-((5-(4-cyanophenoxy)pentyl)oxy)picolyl nitrile (0.2 g, 0.65 mmol 1 eq) was added and the reaction mixture was stirred at room temperature for 15 minutes. Next, the reaction mixture was stirred under reflux for 18 hours. The reaction mixture was cooled to room temperature and methanol (5 mL) was added thereto under ice-cooling conditions, and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was diluted with 1N HCl (20 mL) and washed with ethyl acetate (20 mL). The aqueous layer was basified with 1N NaOH solution (15 mL) and extracted with ethanol-ethyl acetate (20%, 3 × 20 mL). The separated organic layer was dried over anhydrous Na 2 SO 4 and concentrated under vacuum to obtain a crude product, which was purified by reverse-phase HPLC to obtain 5-(5-(4-carbamimidoyl phenoxy)pentyl oxy)picolyl imidoamide as the free base. The free base material was dissolved in 1.25N HCl in ethanol (5 mL). Removal of ethanol under reduced pressure gave a solid, and after lyophilization, white solid 5-(5-(4-carbamimidoyl phenoxy)pentyl oxy)picolyl imidoamide was obtained as the dihydrochloride (0.18 g, 84.68%). Analysis data LCMS: 342 [M+1] + 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.35 (s, 2H), 9.18 (s, 2H), 9.10 (s, 2H), 8.80 (s, 2H), 8.48 (s, 1H), 8.30 (d, 1H), 7.80 (d, 2H), 7.74 (d, 1H), 7.18 (d, 2H), 4.18 (t, 2H), 4.22 (t, 2H), 1.83 (m, 4H), 1.61 (m, 2H).

[0170] Example 2: Preparation of 6-((5-(4-carbamimidoyl phenoxy)pentyl)oxy)nicotinimidamide [Chemical]

[0171] Project 1 To a stirred suspension of NaH (0.28 g, 7.21 mmol, 2.0 equiv) in DMF (6 mL) was added pentane-1,5-diol (0.37 g, 3.62 mmol, 1 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 15 minutes. Next, 6-chloronicotinonitrile (0.35 g, 2.52 mmol, 1 equiv) was added to the reaction mixture. The reaction mixture was stirred at 0 °C for 3 hours. The progress of the reaction was monitored by TLC and LCMS. After the starting material was consumed, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 × 50 mL). The separated organic layer was dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure to obtain the crude product. The crude product was purified by combiflash chromatography to give 6-((5-hydroxypentyl)oxy)nicotinonitrile (0.50 g, 67.02%), which was used in the next step. Analysis data LCMS: 207 [M+1] +

[0172] Project 2 To a stirred solution of 6-((5-hydroxypentyl)oxy)nicotinonitrile (0.15 g, 0.72 mmol, 1.0 equiv) in DMF (5 mL) was added NaH (0.005 g, 1.45 mmol), and the mixture was stirred at 0 °C for 10 minutes. To this mixture was added 4-fluorobenzonitrile (0.10 g, 0.87 mmol, 1.2 equiv) at 0 °C, and the reaction mixture was stirred at room temperature for 2 hours. The progress of the reaction was monitored by TLC and LCMS. After the starting material was consumed, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure to obtain the crude product 6-((5-(4-cyanophenoxy)pentyl)oxy)nicotinonitrile (0.18 g, 80.71%), which was used in the next step without further purification. Analysis data LCMS: 308 [M+1] +

[0173] Step 3 NH in toluene (5 mL) 4 To a stirred suspension of Cl (0.19 g, 3.66 mmol, 8.0 eq) in toluene (5 mL) at 0 °C was added trimethylaluminum (1.83 mL, 3.66 mmol, 8.0 eq). The reaction mixture was stirred at 0 °C for 10 minutes and then at room temperature for 15 minutes. To this solution was added 6-((5-(4-cyanophenoxy)pentyl)oxy)nicotinonitrile (0.14 g, 0.45 mmol, 1.0 eq), and the reaction mixture was stirred at room temperature for 15 minutes. Next, the reaction mixture was stirred under reflux for 18 hours. The reaction mixture was cooled to room temperature, methanol (5 mL) was added under ice-cooling conditions, and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was diluted with 1N HCl (20 mL) and washed with ethyl acetate (20 mL). The aqueous layer was basified with 1N NaOH solution (15 mL) and extracted with ethanol-ethyl acetate (3 × 20 mL, 20%). The separated organic layer was dried over anhydrous Na 2 SO 4 and concentrated under vacuum to obtain a crude product (0.11 g), which was purified by reverse-phase HPLC to obtain 6-((5-(4-carbamimidoylphenoxy)pentyl)oxy)nicotinimidamide as the di-formate salt. The solid was dissolved in 1.25 M HCl in ethanol (8 mL), the solvent was evaporated under reduced pressure to obtain a solid, which was freeze-dried to obtain 6-((5-(4-carbamimidoylphenoxy)pentyl)oxy)nicotinimidamide (0.02 g, 11.17%) as the dihydrochloride salt. Analysis data LCMS: 342 [M+1] + 11H NMR (400 MHz, DMSO-d6) δ 9.30 (brs, 2H), 9.18 (brs, 2H), 8.95 (brs, 2H), 8.78 (brs, 2H), 8.62 (s, 1H), 8.10 (d, 1H), 7.80 (d, 2H), 7.18 (d, 2H), 7.03 (d, 1H), 4.40 (t, 2H), 4.11 (t, 2H), 1.70 - 1.90 (m, 4H), 1.50 - 1.65 (m, 2H).

[0174] Example 3: Preparation of 5 - ((5 - (4 - carbamimidoyl phenoxy) pentyl) oxy) pyrimidine - 2 - carboximidamide

Chemical formula

[0175] Step 1 To a stirred solution of 4 - hydroxybenzonitrile (10 g, 0.08 mol, 1 equiv) in acetone (120 mL), 1,5 - dibromopentane (95.72 g, 0.42 mol, 5.0 equiv) and potassium carbonate (23.21 g, 0.16 mol, 2.0 equiv) were added. The reaction mixture was stirred at 70 °C for 2 h and monitored by TLC and LC - MS. The reaction mixture was diluted with water (2 × 500 mL) and extracted with EtOAC (2 × 800 mL). The separated organic layer was dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure to obtain the crude product. The crude product was purified by a glass column to give 4 - ((5 - bromopentyl) oxy) benzonitrile (15 g, 66.94%), which was used in the next step without further purification. Analysis data LCMS: 268 [M + 1]] +

[0176] Step 2 To a stirred solution of 5-hydroxypyrimidine-2-carbonitrile (0.3 g, 32.47 mmol, 1.0 eq) in acetone (10 mL), a compound of 4-((5-bromopentyl)oxy)benzonitrile (0.79 g, 2.97 mmol, 1.2 eq) and potassium carbonate (0.68 g, 4.95 mmol, 2.0 eq) was added at 70 °C for 2 h. The reaction mixture was monitored by TLC and LCMS. The reaction mixture was diluted with water (50 mL) and extracted with EtOAC (3 × 40 mL). The separated organic layer was dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure to obtain the crude product. The crude product was purified by combiflash chromatography to give 5-((5-(4-cyanophenoxy)pentyl)oxy)pyrimidine-2-carbonitrile (0.2 g, 32.76%) which was used in the next step without further purification. Analysis data LCMS: 309 [M+1] +

[0177] Step 3 To a stirred suspension of NH 4 Cl (0.34 g, 6.49 mmol, 8.0 eq) in toluene (5 mL) at 0 °C, trimethylaluminum (3.2 mL, 6.49 mmol, 8.0 eq) was added. The reaction mixture was stirred at 0 °C for 10 min and then at room temperature for 15 min. To this solution, 5-((5-(4-cyanophenoxy)pentyl)oxy)pyrimidine-2-carbonitrile (0.25 g, 0.81 mmol, 1.0 eq) was added and the reaction mixture was stirred at room temperature for 15 min. Next, the reaction mixture was stirred under reflux for 18 h. The reaction mixture was cooled to room temperature and methanol (5 mL) was added thereto under ice-cooling conditions, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was diluted with 1N HCl (20 mL) and washed with ethyl acetate (20 mL). The aqueous layer was basified with 1N NaOH solution (15 mL) and extracted with ethanol-ethyl acetate (3 × 20 mL, 20%). The separated organic layer was dried over anhydrous Na 2 SO 4It was dried and concentrated under vacuum to obtain a crude product (0.260 g). This crude product was purified by reverse-phase HPLC to obtain 4-((5-((3-aminobenzo[d]isoxazol-6-yl)oxy)pentyl)oxy)benzamide as the di-formate salt. The solid was dissolved in 1.25 M HCl in ethanol (8 mL), and the solvent was evaporated under reduced pressure to obtain a solid. After freeze-drying, 5-((5-(4-carbamimidoylphenoxy)pentyl)oxy)pyrimidine-2-carboximidamide (0.01 g, 5.36%) was obtained as the dihydrochloride salt. Analysis data LCMS: 343 ([M+1]) + 1 1H NMR (400 MHz, DMSO-d 6 ) δ 9.53 (br.s., 2H), 9.30 (br.s., 2H), 9.15 (br.s., 2H), 8.82 (s, 2H), 8.78 (br.s., 2H), 7.82 (d, J = 8.33 Hz, 2H), 7.16 (d, J = 8.33 Hz, 2H), 4.35 (t, J = 6.14 Hz, 2H), 4.13 (t, J = 6.36 Hz, 2H), 1.76 - 1.91 (m, 4H), 1.61 (br.s., 2H)

[0178] Example 4: Preparation of 5-((5-(4-carbamimidoylphenoxy)pentyl)oxy)pyrazine-2-carboximidamide

Chemical formula

[0179] Step 1 A solution of pentane-1,5-diol (0.86 g, 8.26 mmol, 1 equiv) in THF (10 mL) was added to NaH (0.33 g, 8.26 mmol, 1 equiv), and the reaction mixture was stirred at 0 °C for 20 min. 4-Fluorobenzonitrile (1 g, 8.26 mmol, 1 equiv) was added to this solution, and the reaction mixture was stirred at 60 °C for 2 h. The progress of the reaction was monitored by TLC. After the starting material was consumed, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (2 × 200 mL). The combined organic layers were washed with water (3 × 50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 4-((5-hydroxypentyl)oxy)benzonitrile (0.8 g, 47.33%). Analysis data LCMS: 206 [M+1] +

[0180] Step 2 To a solution of 4-((5-hydroxypentyl)oxy)benzonitrile (0.5 g, 2.43 mmol, 1 equiv) in THF (10 mL) was added NaH (0.117 g, 2.92 mmol, 1.2 equiv). The reaction mixture was stirred at 0 °C for 20 min, then 5-chloropyrazine-2-carbonitrile (0.306 g, 2.19 mmol, 0.9 equiv) was added, and the reaction mixture was stirred at room temperature for 2 h. The progress of the reaction was monitored by TLC. After the starting material was consumed, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (2 × 200 mL). The combined organic layers were washed with water (3 × 50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 5-((5-(4-cyanophenoxy)pentyl)oxy)pyrazine-2-carbonitrile (0.4 g, 53.26%). Analysis data LCMS: 309 [M+1] +

[0181] Step 3 A suspension of ammonium chloride (555 mg, 10.39 mmol, 8 equiv) in toluene (5 mL) was added dropwise with trimethylaluminum (5.2 mL, 10.39 mmol, 8 equiv) at 0 °C. The mixture was stirred at the same temperature for 10 minutes and then at room temperature for 15 minutes. To this mixture was added 5-((5-(4-cyanophenoxy)pentyl)oxy)pyrazine-2-carbonitrile (400 mg, 1.30 mmol, 1 equiv), and the reaction mixture was stirred at room temperature for 15 minutes. Next, the reaction mixture was stirred under reflux for 18 hours. The reaction mixture was cooled to room temperature, diluted with methanol (5 mL), and stirred at room temperature for 30 minutes. The reaction mixture was diluted with 3M aqueous HCl (25 mL) and washed with ethyl acetate (20 mL). The aqueous layer was basified with 5N NaOH (20 mL) and extracted with a 1:5 mixture of ethanol-ethyl acetate (3 × 25 mL). The combined organic layers were dried over anhydrous sodium sulfate. Removal of the solvent gave a crude material, which was purified by reverse-phase HPLC to give 5-((5-(4-carbamimidoyl phenoxy)pentyl)oxy)pyrazine-2-carboximidamide as the free base. The solid was dissolved in 1.25M HCl (5 mL), the solution was concentrated under vacuum, and lyophilized to give 5-((5-(4-carbamimidoyl phenoxy)pentyl)oxy)pyrazine-2-carboximidamide as the dihydrochloride (50 mg, 11.26%). Analysis data LCMS: 343 ([M+1]) + 1 H NMR (400 MHz, DMSO-d 6 ) 9.45 (bs, 2H), 9.20 (bs, 2H), 9.15 (bs, 2H), 9.07 (s, 1H), 8.80 (bs, 2H), 8.49 (s, 1H), 7.80 (d, 4H), 7.17 (d, 4H), 4.42 (t, 2H), 4.08 (t, 4H), 1.70 - 1.90 (m, 4H), 1.50 - 1.65 (m, 2H).

[0182] Example 5: Preparation of 5-(4-(4-carbamimidoyl phenoxy)butoxy)picolimidamide

Chemical formula

[0183] Project 1 To a solution of 4-hydroxybenzonitrile (1 g, 8.39 mmol, 1 equiv) in acetone (10 mL) was added K 2 CO 3 (2.32 g, 16.78 mmol, 2 equiv) and 1,4-dibromobutane (7.25 g, 33.56 mmol, 4 equiv), and the reaction mixture was stirred under reflux for 2 h. The progress of the reaction was monitored by TLC. After the starting material was consumed, the reaction mixture was extracted with ethyl acetate (2 × 200 mL). The combined organic layers were washed with water (3 × 50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude material was purified by combiflash using ethyl acetate - hexane to give 4-(4-bromobutoxy)benzonitrile (1.5 g, 70.42%). Analysis data LCMS: 255 [M + 1] +

[0184] Project 2 To a solution of 6-hydroxynicotinonitrile (0.2 g, 1.65 mmol, 1 equiv) in acetone (10 mL) was added K 2 CO 3 (0.57 g, 4.162 mmol, 2.5 equiv) and 4-(4-bromobutoxy)benzonitrile (0.50 g, 1.99 mmol, 1.2 equiv), and the reaction mixture was stirred at 70 °C for 2 h. The progress of the reaction was monitored by TLC. The reaction mixture was cooled to room temperature, diluted with water (150 mL), and stirred at room temperature for 30 min. The precipitate was filtered and dried under vacuum to give 5-(4-(4-cyanophenoxy)butoxy)picolinonitrile (0.3 g, 61.47%), which was used in the next step without further purification. Analysis data LCMS: 294 [M + 1] +

[0185] Project 3 A suspension of ammonium chloride (370 mg, 6.81 mmol, 8 eq) in toluene (8 mL) was added dropwise with trimethylaluminum (3.41 mL, 6.81 mmol, 8 eq) at 0 °C. The mixture was stirred at the same temperature for 10 minutes and then at room temperature for 15 minutes. To this mixture was added 5-(4-(4-cyanophenoxy)butoxy)picolylnitrile (250 mg, 0.85 mmol), and the reaction mixture was stirred at room temperature for 15 minutes. Next, the reaction mixture was stirred under reflux for 18 hours. The reaction mixture was cooled to room temperature, diluted with methanol (5 mL), and stirred at room temperature for 30 minutes. The reaction mixture was diluted with 3M aqueous HCl (25 mL) and washed with ethyl acetate (20 mL). The aqueous layer was basified with 5N NaOH (20 mL) and extracted with a 1:5 mixture of ethanol-ethyl acetate (3 × 25 mL). The combined organic layers were dried over anhydrous sodium sulfate. Removal of the solvent gave a crude product, which was purified by reverse-phase HPLC to give 5-(4-(4-carbamimidoylphenoxy)butoxy)picolylimidamide as the free base. The solid was dissolved in 1.25M HCl (5 mL), the solution was concentrated under vacuum, and lyophilized to give 5-(4-(4-carbamimidoylphenoxy)butoxy)picolylimidamide as the dihydrochloride (80 mg, 28.77%). Analysis data LCMS: 328 [M+1] 1 H NMR (400 MHz, DMSO-d6) δ 9.41 (brs, 2H), 9.20 (brs, 2H), 9.16 (brs, 2H), 8.85 (brs, 2H), 8.46 (s, 1H), 8.32 (d, 1H), 7.82 (d, 2H), 7.71 (d, 1H), 7.18 (d, 2H), 4.20 - 4.28 (m, 2H), 4.10 - 4.19 (m, 2H), 1.85 - 1.96 (m, 4H).

[0186] Example 6: Preparation of 5-(4-(4-carbamimidoylphenoxy)butoxy)picolylimidamide

Chemical formula

[0187] Project 1 To a solution of 4-hydroxybenzonitrile (1.0 g, 8.40 mmol, 1 equiv) in acetone (20 mL) under an inert atmosphere, K 2 CO 3 (2.3 g, 16.80 mmol, 2 equiv) and (E)-1,4-dibromobut-2-ene (5.4 g, 25.21 mmol, 3 equiv) were sequentially added at room temperature. The resulting mixture was stirred at reflux temperature for 4 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (3 × 300 mL). The organic layer was dried over anhydrous sodium sulfate. Removal of the solvent under reduced pressure gave a solid, which was triturated with ether and pentane to give (E)-4-((4-bromobut-2-en-1-yl)oxy)benzonitrile (1.68 g, 80%). Analysis data LCMS: 253 [M+1] +

[0188] Project 2 To a solution of 5-hydroxypicolinonitrile (0.2 g, 1.66 mmol, 1 equiv) in acetone (20 mL) under an inert atmosphere, K 2 CO 3 (0.46 g, 3.2 mmol, 2 equiv) and (E)-4-((4-bromobut-2-en-1-yl)oxy)benzonitrile (0.5 g, 1.99 mmol, 0.5 equiv) were sequentially added at room temperature. The resulting mixture was stirred at reflux temperature for 4 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (3 × 100 mL). The organic layer was dried over anhydrous sodium sulfate. Removal of the solvent under reduced pressure gave a solid, which was triturated with ether and pentane to give (E)-5-((4-(4-isocyanophenoxy)but-2-en-1-yl)oxy)picolinonitrile (0.25 g, 51%). Analysis data LCMS: 292 [M+1] +

[0189] Project 3 A suspension of ammonium chloride (0.54 g, 10.13 mmol, 8 eq) in toluene (5 mL) was added dropwise with trimethylaluminum (5 mL, 10.13 mmol, 8 eq) at 0 °C. The mixture was stirred at the same temperature for 10 minutes and then at room temperature for 15 minutes. To this mixture was added (E)-5-((4-(4-isocyanophenoxy)but-2-en-1-yl)oxy)picolyl nitrile (0.37 g, 1.267 mmol, 1 eq), and the reaction mixture was stirred at room temperature for 15 minutes. Next, the reaction mixture was stirred under reflux for 18 hours. The reaction mixture was cooled to room temperature, diluted with methanol (5 mL), and stirred at room temperature for 30 minutes. The reaction mixture was diluted with 3M aqueous HCl (20 mL) and washed with ethyl acetate (20 mL). The aqueous layer was basified with 5N NaOH solution (15 mL) and extracted with ethanol-ethyl acetate (20%, 3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate. Removal of the solvent gave a crude product, which was purified by reverse-phase HPLC to give 5-(4-(4-carbamimidoylphenoxy)butoxy)picolyl amidine as the free base. The solid was dissolved in 1.25M HCl (5 mL), the solution was concentrated under vacuum, and lyophilized to give 5-(4-(4-carbamimidoylphenoxy)butoxy)picolyl amidine dihydrochloride (0.05 g, 12.07%). Analysis data LCMS: 326 [M+1] 1 H NMR (400 MHz, DMSO-d6) δ 9.40 (brs, 2H), 9.08 (brs, 2H), 8.82 (brs, 2H), 8.53 (brs, 2H), 8.30 (d, 1H), 7.82 (d, 2H), 7.74 (d, 1H), 7.19 (d, 2H), 6.17 (brs, 2H), 4.82 (brs, 2H), 4.74 (brs, 2H).

[0190] Example 7: Preparation of 5,5'-(pentane-1,5-diylbis(oxy))bis(pyrazine-2-carboximidamide)

Chemical formula

[0191] Project 1 To a stirred solution of pentane-1,5-diol (0.22 g, 2.15 mmol, 1 equiv) in THF (10 mL) at 0 °C was added NaH (0.26 g, 6.44 mmol, 3.0 equiv), and the reaction mixture was stirred at the same temperature for 10 min. To this solution was added 5-chloropyrazine-2-carbonitrile (0.30 g, 2.14 mmol, 3.0 equiv), and the resulting mixture was stirred at room temperature for 3 h. The progress of the reaction was monitored by TLC and LCMS. After completion, the reaction mixture was diluted with water (50 ml) and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure to give the crude product. The crude product was purified by combiflash on silica gel using an ethyl acetate-hexane system as the eluent to give 5,5'-(pentane-1,5-diylbis(oxy))bis(pyrazine-2-carbonitrile) (0.36 g, 53.81%). Analytical data LCMS: 311 [M+1]] +

[0192] Project 2 To a stirred suspension of NH 4 Cl (1.10 g, 20.64 mmol, 16 equiv) in toluene (10 mL) at 0 °C was added trimethylaluminum (10.32 mL, 20.64 mmol, 16 equiv). The reaction mixture was stirred at 0 °C for 10 min and then at room temperature for 15 min. To this solution was added 5,5'-(pentane-1,5-diylbis(oxy))bis(pyrazine-2-carbonitrile) (0.40 g, 1.29 mmol, 1.0 equiv), and the reaction mixture was stirred at room temperature for 15 min. Next, the reaction mixture was stirred under reflux for 18 h. The reaction mixture was cooled to room temperature, methanol (5 mL) was added under ice-cooling conditions, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was diluted with 1N HCl (20 mL) and washed with ethyl acetate (20 mL). The aqueous layer was basified with 1N NaOH solution (15 mL) and extracted with ethanol-ethyl acetate (20%, 3 × 20 mL). The separated organic layers were dried over anhydrous Na 2 SO4 It was dried and concentrated under vacuum to obtain a crude product (0.35 g). This crude product was purified by reverse-phase HPLC to obtain 5,5’-(pentane-1,5-diylbis(oxy))bis(pyrazine-2-carboximidamide) 5-5-(hepta-1,6-diyne-1,7-diyl) as the di-formate. The solid was dissolved in 1.25 M HCl in ethanol (8 mL), and then the solvent was evaporated under reduced pressure. The obtained solid was freeze-dried to obtain 5,5’-(pentane-1,5-diylbis(oxy))bis(pyrazine-2-carboximidamide) as the dihydrochloride (0.08 g, 13.25%). Analysis data LCMS: 345 [M+1] + 1 1H NMR (400 MHz, DMSO-d 6 ) δ 9.52 (brs, 4H), 9.24 (brs, 4H), 9.05~9.12 (brs, 2H), 8.52 (brs, 2H), 4.47 (t, 4H), 1.92 - 1.80 (m, 4H), 1.55~1.65 (m, 2H)

[0193] Example 8: Preparation of 6,6’-(heptane-1,7-diyl)dipicolylimidamide

Chemical formula

[0194] Step 1 To a stirred solution of hepta-1,6-diyne (0.30 g, 3.26 mmol, 1.0 equiv) in THF (20 mL), 6-bromopicolinonitrile (1.8 g, 9.38 mmol, 3.0 equiv), triethylamine (1.37 mL, 9.38 mmol, 3.0 equiv) and CuI (62 mg, 0.32 mmol, 0.1 equiv) were added. The resulting reaction mixture was deoxygenated by purging with nitrogen for 20 minutes. To this mixture, (Ph 3 P) 4Pd (0.188 g, 0.163 mmol, 0.05 eq) was added, and the reaction mixture was deoxygenated by purging with nitrogen again for 10 minutes. The reaction mixture was stirred at 60 °C for 4 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with brine, dried over sodium sulfate, and evaporated under reduced pressure to obtain a crude product, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to obtain 6,6'-(hepta-1,6-diyne-1,7-diyl)dipicolinonitrile (400 mg, 40.40%). Analysis data LCMS: 297 [M+1] +

[0195] Step 2 To a stirred suspension of 6,6'-(hepta-1,6-diyne-1,7-diyl)dipicolinonitrile (0.3 g, 1.01 mmol) in a solution of ethyl acetate (10 mL) and methanol (10 mL) was added Pt / O 2 (40 mg). The reaction mixture was stirred at room temperature for 2 hours under a hydrogen atmosphere. The progress of the reaction was monitored by TLC and 1 1H NMR. After completion, the reaction mixture was filtered through a Celite pad, and the pad was washed with ethyl acetate (20 mL). The filtrate was evaporated under reduced pressure to obtain a crude product, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to obtain 6,6'-(heptane-1,7-diyl)dipicolinonitrile (250 mg, 82.50%). Analysis data LCMS: 305 [M+1] +

[0196] Step 3 NH in toluene (8 mL) 4To a stirred suspension of Cl (0.32 g, 6.052 mmol, 8 eq) at 0 °C, a 2 M solution of trimethylaluminum in toluene (3 mL, 6.052 mmol, 8 eq) was added. The reaction mixture was stirred at 0 °C for 10 minutes and then at room temperature for 15 minutes. To this solution, 6,6’-(heptane-1,7-diyl)dipicolinonitrile (0.230 g, 0.75 mmol, 1.0 eq) was added and the reaction mixture was stirred at room temperature for 15 minutes. Next, the reaction mixture was stirred under reflux for 18 hours. The reaction mixture was cooled to room temperature, methanol (5 mL) was added under ice-cooling conditions and then stirred at room temperature for 30 minutes. The reaction mixture was diluted with 1 N HCl (20 mL) and washed with ethyl acetate (20 mL). The aqueous layer was basified with 1 N NaOH solution (15 mL) and extracted with ethanol-ethyl acetate (20%, 3 × 20 mL). The separated organic layer was dried over anhydrous Na 2 SO 4 and concentrated under vacuum to give a crude product (0.3 g). This crude product was purified by reverse-phase HPLC to give 6,6’-(heptane-1,7-diyl)dipicolinimide amide as the di-formate salt. The solid was dissolved in 1.25 M HCl in ethanol (8 mL), the solvent was evaporated under reduced pressure, and the resulting material was lyophilized to give 6,6’-(heptane-1,7-diyl)dipicolinimide amide as the dihydrochloride salt (0.06 g, 21.89%). Analysis data LCMS: 339 [M+1] + 1 H NMR (400 MHz, DMSO-d6) δ 9.50 (brs, 8H), 8.20 (d, 2H), 8.04 (t, 2H), 7.62 (d, 2H), 6.60 (brs, 2H), 2.82 (t, 4H), 1.78 - 1.63 (m, 4H), 1.40 - 1.20 (m, 6H).

[0197] Example 9: Preparation of 5,5’-(heptane-1,7-diyl)dinicotinimide amide

Chemical formula

[0198] Step 1 A stirred solution of hepta-1,6-diyne (0.1 g, 1.089 mmol, 1.0 equiv) in THF (10 mL) was added with methyl 5-bromonicotinate (0.69 g, 3.62 mmol, 3.0 equiv), triethylamine (0.45 mL, 3.26 mmol, 3.0 equiv), and CuI (20 mg, 0.108 mmol, 0.1 equiv). The resulting reaction mixture was deoxygenated by purging with N 2 for 20 min. Next, Pd(PPh 3 ) 4 (62 mg, 0.0544 mmol, 0.05 equiv) was added and the reaction mixture was deoxygenated again by purging with nitrogen for 10 min. The reaction mixture was stirred at 70 °C for 4 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with brine (20 mL), dried over sodium sulfate, and evaporated under reduced pressure to give a crude material, which was purified by silica gel column chromatography using an ethyl acetate-hexane system as the eluent to afford 5,5’-(hepta-1,6-diyne-1,7-diyl)dinicotinate dimethyl (180 mg, 45.80%). Analytical data LCMS: 343.4 [M+1] +

[0199] Step 2 Pd-C (150 mg) was added to a stirred suspension of 5,5’-(hepta-1,6-diyne-1,7-diyl)dinicotinate dimethyl (0.18 g, 0.593 mmol) in methanol (5 mL). The reaction mixture was stirred at room temperature for 2 h under a hydrogen atmosphere. The progress of the reaction was monitored by TLC and 1 1H NMR. After completion of the reaction, the mixture was filtered through a Celite pad, the pad was washed with ethyl acetate (20 mL), and the filtrate was evaporated under reduced pressure to give a crude material, which was purified by silica gel column chromatography using an ethyl acetate-hexane system as the eluent to afford 5,5’-(heptane-1,7-diyl)dinicotinate dimethyl (120 mg, 65.57%).

[0200] Step 3 NH in toluene (5 mL) 4 To a stirred suspension of NH Cl (0.15 g, 6.052 mmol, 8 eq) in toluene (5 mL) at 0 °C was added a 2 M solution of trimethylaluminum in toluene (1.45 mL, 2.909 mmol, 8 eq). The reaction mixture was stirred at 0 °C for 10 minutes and then at room temperature for 15 minutes. To this solution was added 5,5'-(heptane-1,7-diyl)dinicotinate dimethyl (0.12 g, 0.363 mmol, 1.0 eq), and the reaction mixture was stirred at room temperature for 15 minutes. Next, the reaction mixture was stirred under reflux for 18 hours. The reaction mixture was cooled to room temperature, methanol (3 mL) was added under ice-cooling conditions, and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was diluted with 1 N HCl (20 mL) and washed with ethyl acetate (20 mL). The aqueous layer was basified with 1 N NaOH solution (15 mL) and extracted with ethanol-ethyl acetate (20%, 3 × 20 mL). The separated organic layer was dried over anhydrous Na 2 SO 4 and concentrated under vacuum to give a crude product (0.3 g). This crude product was purified by reverse-phase HPLC to give 5,5'-(heptane-1,7-diyl)dinicotinimide amide as the di-formate. The solid was dissolved in 1.25 M HCl in ethanol (8 mL), the solvent was evaporated under reduced pressure to give a solid, and this was lyophilized to give 5,5'-(heptane-1,7-diyl)dinicotinimide amide as the dihydrochloride (0.022 g, 14.59%). Analysis data LCMS: 339.3 [M+1] + 1 H NMR (400 MHz, DMSO-d6) δ 9.55 (brs, 4H), 9.25 (brs, 4H), 8.81 (s, 2H), 8.75 (s, 2H), 8.08 (s, 2H), 2.70 - 2.60 (m, 4H), 1.70 - 1.55 (m, 4H), 1.40 - 1.22 (m, 6H).

[0201] Example 10: Preparation of 6,6'-(heptane-1,7-diyl)dinicotinimide amide

Chemical formula

[0202] Project 1 To a stirred solution of hepta-1,6-diyne (0.25 g, 2.71 mmol, 1.0 equiv) in THF (20 mL) were added methyl 6-bromonicotinate (1.46 g, 6.79 mmol, 2.5 equiv), triethylamine (1.14 mL, 8.13 mmol, 3.0 equiv) and CuI (52 mg, 0.271 mmol, 0.1 equiv), and the resulting mixture was deoxygenated by purging with N 2 for 20 min. To this mixture was added Pd(PPh 3 ) 4 (156 mg, 0.135 mmol, 0.05 equiv), and the reaction mixture was deoxygenated again by purging with N 2 for 10 min. The reaction mixture was stirred at 60 °C for 4 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, diluted with water (30 mL), and extracted with ethyl acetate (3 × 35 mL). The combined organic layers were washed with brine (30 mL), dried over sodium sulfate, and evaporated under reduced pressure to give a crude product, which was purified by silica gel column chromatography using an ethyl acetate–hexane system as the eluent to give 6,6’-(hepta-1,6-diyne-1,7-diyl)dinicotinate dimethyl (0.95 g, 96.93%). Analysis data LCMS: 363.3 [M+1] +

[0203] Project 2 To a stirred suspension of 6,6’-(hepta-1,6-diyne-1,7-diyl)dinicotinate dimethyl (0.95 g, 2.76 mmol) in methanol (15 mL) and ethyl acetate (5 mL) was added Pd-C (1 g). The reaction mixture was stirred at room temperature under a hydrogen atmosphere for 2 h. TLC and 1The progress of the reaction was monitored by ¹H NMR. After completion, the mixture was filtered through a Celite pad, and the pad was washed with ethyl acetate (50 mL). The filtrate was evaporated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography using an ethyl acetate - hexane system as the eluent to obtain 6,6'-(heptane-1,7-diyl)dinicotinate dimethyl (0.6 g, 61.79%).

[0204] Step 3 NH 4 Cl (0.19 g, 3.51 mmol, 10 equiv) in toluene (7 mL) was added to a stirred suspension at 0 °C, and a 2 M solution of trimethylaluminum in toluene (1.75 mL, 3.51 mmol, 10 equiv) was added. The reaction mixture was stirred at 0 °C for 10 minutes and then at room temperature for 15 minutes. To this solution, 6,6'-(heptane-1,7-diyl)dinicotinate dimethyl (0.13 g, 0.351 mmol, 1.0 equiv) was added, and the reaction mixture was stirred at room temperature for 15 minutes. Next, the reaction mixture was stirred under reflux for 18 hours. The reaction mixture was cooled to room temperature, methanol (5 mL) was added under ice-cooling conditions, and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was diluted with 1 N HCl (30 mL) and washed with ethyl acetate (20 mL). The aqueous layer was basified with 1 N NaOH solution (25 mL) and extracted with ethanol - ethyl acetate (20%, 3 × 60 mL). The separated organic layer was dried over anhydrous Na 2 SO 4 and concentrated under vacuum to obtain a crude product (0.3 g), which was purified by reverse-phase HPLC to obtain 5,5'-(heptane-1,7-diyl)dinicotinimide amide as the di-formate salt. The solid was dissolved in 1.25 M HCl in ethanol (8 mL), the solvent was evaporated under reduced pressure to obtain a solid, which was lyophilized to obtain 6,6'-(heptane-1,7-diyl)dinicotinimide amide as the dihydrochloride salt (0.012 g, 110.16%). Analysis data LCMS: 339.2 [M+1] + 1 ¹H NMR (400 MHz, DMSO-d 6)δ 9.30 - 9.44 (brs, 4H), 8.97 - 9.13 (brs, 4H), 8.81 - 8.92 (s, 2H), 8.01 - 8.23 (m, 2H), 7.39 - 7.63 (m, 2H), 2.75 - 2.93 (m, 4H), 1.60 - 1.78 (m, 4H), 1.23 - 1.46 (m, 6H).

[0205] Example 11: Preparation of 5-(5-(3-carbamimidoylphenoxy) pentyloxy) picolinimidamide [Chemical formula]

[0206] Step 1 To a stirred solution of 3-hydroxybenzonitrile (2.0 g, 16.78 mmol, 1.0 equivalent) in acetone (20 mL), 1,5-dibromopentane (11.58 g, 3.0 equivalents) and K 2 CO 3 (4.41 g, 31.95 mmol, 2 equivalents) were added, and the reaction mixture was stirred at 80 °C for 2 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was filtered and the solid was washed with acetone (20 mL). Removal of acetone under reduced pressure gave an oily residue, which was purified by silica gel column chromatography using an ethyl acetate - hexane system as the eluent to give 3-(5-bromopentyloxy) benzonitrile (3 g, 66.66%). Analysis data LCMS: 268 [M + 1] +

[0207] Step 2 To a stirred solution of 5-hydroxypyridine-2-carbonitrile (0.2 g, 1.66 mmol, 1 equivalent) in DMF (10 mL), 3-(5-bromopentyloxy) benzonitrile (0.49 g, 1.83 mmol, 1.1 equivalents) and K 2 CO 3(0.343 g, 2.49 mmol, 1.5 equiv) was added, and the reaction mixture was stirred at 80 °C for 2 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 × 150 mL). The combined organic layers were dried over sodium sulfate, filtered, and evaporated to dryness under vacuum to obtain a residue, which was purified by combi-flash on silica gel using an ethyl acetate - hexane system as the eluent to give 5-(5-(3-cyanophenoxy) pentyloxy) picolinonitrile (0.3 g, 58.61%). Analysis data LCMS: 308 [M + 1] +

[0208] Step 3 NH in toluene (10 mL) 4To a stirred suspension of Cl (0.522 g, 9.76 mmol, 10 eq) at 0 °C, trimethylaluminum (4.88 mL, 9.76 mmol, 10 eq) was added dropwise under nitrogen. The reaction mixture was stirred at 0 °C for 10 minutes and then at room temperature for 15 minutes. To this mixture, 5-{[5-(4-cyanophenoxy)pentyl]oxy}pyridine-2-carbonitrile (1.0 g, 3.25 mmol, 1.0 eq) was added at 0 °C, and the reaction mixture was stirred at room temperature for 15 minutes. The reaction mixture was stirred at 120 °C for 18 hours. The reaction mixture was cooled to room temperature, quenched by dropwise addition of methanol (5 mL) at 0 °C, and then stirred at room temperature for 30 minutes. The reaction mixture was acidified with 3 M aqueous HCl (50 mL) and extracted with ethyl acetate (20 mL). The organic layer was separated, the aqueous layer was basified using 5 N NaOH solution (50 mL), and extracted with 20% ethanol-ethyl acetate (3 × 200 mL). The combined organic layers were dried over sodium sulfate, filtered, and evaporated to dryness. The residue was dried completely with a toluene azeotrope. The residue was triturated with 50% ethanol-ethyl acetate mixture (2 × 50 mL), and the solid was removed by filtration. The filtrate was evaporated to give the crude product, which was purified by reverse-phase HPLC to give the free base of 5-(5-(3-carbamimidoylphenoxy)pentyl)picolinimidamide. This solid was dissolved in 1.25 M HCl in ethanol (5 mL) at 0 °C, the ethanol was removed, and the residue was lyophilized to give 5-(5-(3-carbamimidoylphenoxy)pentyl)picolinimidamide dihydrochloride (130 mg, 39%). Analysis data LCMS: 342 [M+1] + 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.43 (d, 4H), 9.25 (d, 4H), 8.45 (d, 1H), 8.40 (d, 1H), 7.73 (d, 1H), 7.50 (t, 1H), 7.43 - 7.35 (m, 2H), 7.27 (d, 1H), 4.22 (t, 2H), 4.10 (t, 1H), 1.90 - 1.78 (m, 4H), 1.66 - 1.55 (m, 2H).

[0209] Example 12: Preparation of 4-({5-[(6-Cyanopyridin-3-yl)oxy]pentyl}oxy)pyridine-2-carbonitrile [Chemical formula]

[0210] Step 1 To a stirred solution of pentane-1,5-diol (500 mg, 3.62 mmol, 1.0 equivalent) in DMF (15 mL) at 0 °C was added NaH (217 mg, 5.43 mmol, 1.5 equivalents), and the resulting reaction mixture was stirred at 0 °C for 15 minutes, followed by the addition of 4-chloropyridine-2-carbonitrile (754 mg, 7.24 mmol, 2.0 equivalents). The reaction mixture was stirred at room temperature for 12 hours. The progress of the reaction was monitored by TLC. After 4-chloropyridine-2-carbonitrile was consumed, the reaction mixture was diluted with ethyl acetate (50 mL) and washed with water (3 × 20 mL). The organic layer was dried over sodium sulfate. Removal of ethyl acetate under reduced pressure gave a crude oil, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to give 4-[(5-hydroxypentyl)oxy]pyridine-2-carbonitrile (350 mg, 46.41%).

[0211] Step 2 A stirred solution of 4-[(5-hydroxypentyl)oxy]pyridine-2-carbonitrile (350 mg, 3.62 mmol, 1.0 equiv) in DMF (10 mL) at 0 °C was treated with NaH (102 mg, 2.53 mmol, 1.5 equiv), and the resulting reaction mixture was stirred at 0 °C for 15 min, followed by the addition of 5-fluoropyridine-2-carbonitrile (413 mg, 3.39 mmol, 2.0 equiv). The reaction mixture was stirred at room temperature for 12 h. The progress of the reaction was monitored by TLC. After consumption of 4-[(5-hydroxypentyl)oxy]pyridine-2-carbonitrile, the reaction mixture was diluted with ethyl acetate (50 mL) and washed with water (3 × 20 mL). The organic layer was dried over sodium sulfate and concentrated under reduced pressure to afford a crude oil, which was purified by combiflash on silica gel using an ethyl acetate–hexane system as eluent to give 4-({5-[(6-cyanopyridin-3-yl)oxy]pentyl}oxy)pyridine-2-carbonitrile (185 mg, 35.37%). Analysis data LCMS: 309 [M+1] +

[0212] Step 3 NH in toluene (10 mL) 4To a stirred suspension of Cl (257 mg, 4.80 mmol, 8 eq) at 0 °C, trimethylaluminum (2.40 mL, 4.80 mmol, 8 eq) was added dropwise under nitrogen. The reaction mixture was stirred at 0 °C for 10 minutes and then at room temperature for 15 minutes. The reaction mixture was cooled to 0 °C and 4-({5-[(6-cyanopyridin-3-yl)oxy]pentyl}oxy)pyridine-2-carbonitrile (187 mg, 0.60 mmol, 1 eq) was added. The reaction mixture was stirred at room temperature for 15 minutes and then at 120 °C for 16 hours. The reaction mixture was cooled to 0 °C, methanol (5 mL) was added dropwise, and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was acidified with 2M HCl solution (150 mL) and extracted with ethyl acetate (50 mL). The organic layer was separated, the aqueous layer was basified using 5N NaOH solution (50 mL), and extracted with 20% ethanol-ethyl acetate (5 x 200 mL). The combined organic layers were dried over sodium sulfate and evaporated to dryness. A trace amount of water was removed with a toluene azeotrope to obtain a solid residue. The solid was triturated with 1:1 ethanol-ethyl acetate (2 x 200 mL) and filtered. The filtrate was evaporated to dryness and then the residue was purified by reverse phase HPLC to obtain 4-({5-[(6-cyanopyridin-3-yl)oxy]pentyl}oxy)pyridine-2-carbonitrile as the free base. This solid was dissolved in 1.25M HCl in ethanol (5 mL) at 0 °C, the solvent was evaporated to dryness, and freeze-dried to obtain 4-({5-[(6-cyanopyridin-3-yl)oxy]pentyl}oxy)pyridine-2-carbonitrile hydrochloride (20 mg, 7.00%). Analysis data LCMS: 342 [M+1] + 1 1H NMR (400 MHz, DMSO-d6) δ d9.60 (s, 2H), 9.40 (d, 4H), 9.16 (s., 2H), 8.60 (d, 1H), 8.48 (d, 1H), 8.33 (d, 1H), 7.98 (d, 1H), 7.83 - 7.55 (m, 1H), 7.47 - 7.27 (m, 1H), 4.23 (t, 4H), 1.97 - 1.83 (m, 4H), 1.65 - 1.55 (m, 2H).

[0213] Example 13: Preparation of 5 - ((((1R,4R)-4-(4-carbamimidoyl phenoxy) cyclohexyl) oxy) picolinimidamide [Chemical formula]

[0214] Step 1 To a solution of trans - cyclohexane - 1,4 - diol (1 g, 8.60 mmol, 1.0 equivalent) in DMSO (10 mL) at 0 °C was added NaH (60% in mineral oil) (104 mg, 4.30 mmol, 0.5 equivalent) under an inert atmosphere, and the resulting mixture was stirred at the same temperature for 15 minutes. To this solution was added 4 - fluorobenzonitrile (522 mg, 4.30 mmol, 0.5 equivalent) in DMSO (2 mL), and the resulting reaction mixture was stirred at room temperature for 1 hour. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with ice - cold water (50 ml) and extracted with ethyl acetate (3 × 200 mL). The combined organic layers were washed with water (5 × 100 mL), followed by brine (50 mL), and dried over anhydrous sodium sulfate. Removal of the solvent under reduced pressure gave a crude material, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to give 4 - ((((1R,4R)-4 - hydroxycyclohexyl) oxy) benzonitrile (900 mg, 50%).

[0215] Step 2 Under an inert atmosphere, to a solution of 4-(((1r,4r)-4-hydroxycyclohexyl)oxy)benzonitrile (300 mg, 1.38 mmol, 1.0 eq) in DMSO (5 mL) at 0 °C was added NaH (60% in mineral oil) (49.68 mg, 2.07 mmol, 1.5 eq), and the resulting mixture was stirred at the same temperature for 15 minutes. To this solution was added a solution of 5-fluoropicolinonitrile (202.3 mg, 1.65 mmol, 1.2 eq) in DMSO (2 mL), and the resulting reaction mixture was stirred at room temperature for 1 hour. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with ice-cold water (50 mL) and extracted with ethyl acetate (3 × 150 mL). The combined organic layers were washed with water (5 × 100 mL), followed by brine (50 mL), and dried over anhydrous sodium sulfate. Removal of the solvent under reduced pressure gave a crude material, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to give 5-(((1r,4r)-4-(4-cyanophenoxy)cyclohexyl)oxy)picolinonitrile (200 mg, 45.35%).

[0216] Step 3 To a suspension of ammonium chloride (267.3 mg, 5.0 mmol, 8 eq) in toluene (5 mL), trimethylaluminum (2 M) (721 mg, 2.5 mL, 5.0 mmol, 8 eq) was added dropwise at 0 °C. The mixture was stirred at the same temperature for 10 minutes and then at room temperature for 15 minutes. To this mixture, 5-(((1r,4r)-4-(4-cyanophenoxy)cyclohexyl)oxy)picolyl nitrile (200 mg, 0.62 mmol, 1.0 eq) was added and the reaction mixture was stirred at room temperature for an additional 15 minutes. Next, the reaction mixture was stirred under reflux for 16 hours. The reaction mixture was cooled to room temperature, diluted with methanol (5 mL), and stirred at room temperature for 30 minutes. The reaction mixture was diluted with 1N aqueous HCl (25 mL) and washed with ethyl acetate (50 mL). The aqueous layer was basified with 5N NaOH (20 mL) and extracted with a 1:5 mixture of ethanol-ethyl acetate (3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate. Removal of the solvent gave a crude product, which was purified by reverse-phase HPLC to give 5-(((1r,4r)-4-(4-carbamimidoyl phenoxy)cyclohexyl)oxy)picolyl amidine as the free base. The solid was dissolved in 1.25 M HCl in ethanol (5 mL), the solution was concentrated under vacuum, and lyophilized to give 5-(((1r,4r)-4-(4-carbamimidoyl phenoxy)cyclohexyl)oxy)picolyl amidine dihydrochloride (5 mg, 4.95%). Analysis data LCMS: 354 [M+1] + 1 H NMR (400 MHz, DMSO-d6) δ 8.80 - 8.54 (m, 6H), 8.46 (d, 1H), 8.27 (d, 1H), 7.82 (d, 2H), 7.77 (d, 1H), 7.20 (d, 2H), 4.76 (brs, 1H), 4.69 (brs, 1H), 2.15 - 2.00 (m, 4H), 1.75 - 1.60 (m, 4H).

[0217] Example 14: Preparation of 5-(((1s,4s)-4-(4-carbamimidoyl phenoxy)cyclohexyl)oxy)picolyl amidine [Chemistry]

[0218] Project 1 To a solution of trans-cyclohexane-1,4-diol (300 mg, 2.58 mmol, 1.0 equiv) in DMSO (5 mL) at 0 °C was added NaH (60% in mineral oil) (30.98 mg, 1.29 mmol, 0.5 equiv) under an inert atmosphere, and the resulting mixture was stirred at the same temperature for 15 minutes. To this solution was added 4-fluorobenzonitrile (312.9 mg, 2.58 mmol, 1.0 equiv) in DMSO (2 mL), and the resulting reaction mixture was stirred at room temperature for 1 hour. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with ice-cold water (50 mL) and extracted with ethyl acetate (3 × 200 mL). The combined organic layers were washed with water (5 × 100 mL) and brine, and dried over anhydrous sodium sulfate. Removal of the solvent under reduced pressure gave a crude material, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to give 4-(((1s,4s)-4-hydroxycyclohexyl)oxy)benzonitrile (200 mg, 71.4%).

[0219] Project 2 Under an inert atmosphere, to a solution of 4-(((1s,4s)-4-hydroxycyclohexyl)oxy)benzonitrile (180 mg, 0.82 mmol, 1.0 equiv) in DMSO (5 mL) at 0 °C was added NaH (60% in mineral oil) (29.52 mg, 1.23 mmol, 1.5 equiv), and the resulting mixture was stirred at the same temperature for 15 minutes. To the mixture was added a solution of 5-fluoropicolinonitrile (121.4 mg, 0.99 mmol, 1.2 equiv) in DMSO (2 mL), and the resulting reaction mixture was stirred at room temperature for 1 hour. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with ice-cold water (50 mL) and extracted with ethyl acetate (3 × 150 mL). The combined organic layers were washed with water (5 × 100 mL), brine, and dried over anhydrous sodium sulfate. Removal of the solvent under reduced pressure gave a crude material, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to give 5-(((1s,4s)-4-(4-cyanophenoxy)cyclohexyl)oxy)picolinonitrile (220 mg, 83.3%).

[0220] Step 3 A suspension of ammonium chloride (294.2 mg, 5.5 mmol, 8 equiv) in toluene (6 mL) was added dropwise with trimethylaluminum (2 M) (793.1 mg, 2.75 mL, 5.5 mmol, 8 equiv) at 0 °C. The mixture was stirred at the same temperature for 10 minutes and then at room temperature for 15 minutes. 5-(((1s,4s)-4-(4-Cyanophenoxy)cyclohexyl)oxy)picolyl nitrile (220 mg, 0.68 mmol, 1.0 equiv) was added to this mixture, and the reaction mixture was stirred at room temperature for an additional 15 minutes. Next, the reaction mixture was stirred under reflux for 16 hours. The reaction mixture was cooled to room temperature, diluted with methanol (5 mL), and stirred at room temperature for 30 minutes. The reaction mixture was diluted with 1N aqueous HCl (25 mL) and washed with ethyl acetate (50 mL). The aqueous layer was basified with 5N NaOH (20 mL) and extracted with a 1:5 mixture of ethanol-ethyl acetate (3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate. Removal of the solvent gave a crude product, which was purified by reverse-phase HPLC to give 5-(((1s,4s)-4-(4-carbamimidoyl phenoxy)cyclohexyl)oxy)picolyl imidoamide as the free base. The solid was dissolved in 1.25 M HCl in ethanol (5 mL), the solution was concentrated under vacuum, and lyophilized to give 5-(((1s,4s)-4-(4-carbamimidoyl phenoxy)cyclohexyl)oxy)picolyl imidoamide dihydrochloride (5 mg, 4.95%). Analysis data LCMS: 354 [M+1] + 1 1H NMR (400 MHz, DMSO-d6) δ 11.33 - 10.49 (m, 6H), 8.48 (brs, 1H), 8.22 (brs, 1H), 7.81 - 7.74 (m, 3H), 7.19 (d, 2H), 2.00~1.75 (m, 8H).

[0221] Example 15: Preparation of 4-(5-(3-Carbamimidoyl phenoxy)pentyl oxy)picolyl imidoamide

Chemical formula

[0222] Project 1 To a stirred solution of 1,5-pentanediol (2.25 g, 2.16 mmol, 3.0 equiv) in dimethylformamide (10 mL) was added sodium hydride (1.5 g, 1.08 mmol, 1.5 equiv) at 0 °C. The reaction mixture was stirred at room temperature for 15 minutes. To this mixture was added 4-chloro-pyridine-2-carbonitrile (1.0 g, 7.20 mmol, 1.0 equiv), and the reaction mixture was stirred at room temperature for 15 hours. The progress of the reaction was monitored by TLC. After the 4-chloropyridine-2-carbonitrile was consumed, the reaction mixture was diluted with ethyl acetate (50 mL) and washed with water (3 × 20 mL). The organic layer was dried over sodium sulfate and concentrated under reduced pressure to give a crude oil, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to give 4-[(5-hydroxypentyl)oxy]pyridine-2-carbonitrile (600 mg, 40.54%).

[0223] Project 2 To a stirred solution of 4-(5-hydroxypentyloxy)picolinitrile (0.450 g, 2.18 mmol, 1 equiv) in dichloromethane (5 mL) was added TEA (0.33 g, 3.27 mmol, 1.5 equiv) at 0 °C, and the reaction mixture was stirred at 0 °C for 10 minutes. Next, to this solution was added methanesulfonyl chloride (0.299 g, 2.61 mmol, 1.2 equiv) at 0 °C. The reaction mixture was stirred at room temperature for 60 minutes. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with water (50 mL) and extracted with dichloromethane (3 × 50 mL). The combined organic layers were dried over sodium sulfate and concentrated in vacuo to give a crude residue, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to give 5-(2-cyanopyridin-4-yloxy)pentyl methanesulfonate (0.4 g, 68.4%).

[0224] Project 3 To a stirred solution of 5-(2-cyanopyridin-4-yloxy)pentyl methanesulfonate (0.4 g, 1.49 mmol, 1 equiv) in DMF (5 mL), K 2 CO 3 (0.61 g, 4.47 mmol, 3 equiv) was added at room temperature and the reaction mixture was stirred at room temperature for 10 min. To this mixture was added 3-hydroxybenzonitrile (0.23 g, 1.93 mmol, 1.3 equiv) at room temperature and the reaction mixture was stirred at room temperature for 2 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were dried over sodium sulfate and concentrated to dryness under vacuum to give the residual crude product, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as eluent to give 4-(5-(3-cyanophenoxy) pentyloxy) picolinonitrile (0.2 g, 43.67%). Analysis data LCMS: 308 [M + 1] +

[0225] Step 4 NH in toluene (6 mL) 4To a stirred suspension of Cl (0.348 g, 6.51 mmol, 10 equiv) at 0 °C, trimethylaluminum (3.25 mL, 6.51 mmol, 10 equiv) was added dropwise under nitrogen. The reaction mixture was stirred at 0 °C for 10 minutes and then at room temperature for 15 minutes. To this mixture, 4-(5-(3-cyanophenoxy) pentyloxy) picolinonitrile (0.2 g, 0.65 mmol, 1.0 equiv) was added at 0 °C, and the reaction mixture was stirred at room temperature for 15 minutes. The reaction mixture was stirred at 120 °C for 18 hours. The reaction mixture was cooled to room temperature, quenched by dropwise addition of methanol (5 mL) at 0 °C, and then stirred at room temperature for 30 minutes. The reaction mixture was acidified with 3M HCl aqueous solution (50 mL) and extracted with ethyl acetate (20 mL). The organic layer was separated, the aqueous layer was basified using 5N NaOH solution (50 mL), and extracted with 20% ethanol-ethyl acetate (3 × 200 mL). The combined organic layers were dried over sodium sulfate, filtered, and evaporated to dryness. The residue was completely dried with a toluene azeotrope. The residue was triturated with 50% ethanol-ethyl acetate mixture (2 × 50 mL), and the solid was removed by filtration. The filtrate was evaporated to give the crude product, which was purified by reverse-phase HPLC to give 4-(5-(3-carbamimidoylphenoxy) pentyloxy) picolinimidamide as the free base. This solid was dissolved in 1.25M HCl in ethanol (5 mL) at 0 °C, the ethanol was removed, and the residue was lyophilized to give the dihydrochloride salt of 4-(5-(3-carbamimidoylphenoxy) pentyloxy) picolinimidamide (30 mg, 13.49%). Analysis data LCMS: 342 [M+1] + 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.65 (brs, 2H), 9.43 (s, 2H), 9.47 (s, 2H), 9.26 (brs, 2H), 8.60 (d, 1H), 8.06 (brs, 1H), 7.51 (m, 1H), 7.41 (m, 2H), 7.31 (m, 2H), 4.24 (t, 2H), 4.10 (m, 2H), 1.84 (d, 4H), 1.59 (brs, 2H).

[0226] Example 16: Preparation of 5,5'-(butane-1,4-diylbis(oxy))dipicolinimidamide [Chemical formula]

[0227] Step 1 To a solution of 1,4-dibromobutane (500 mg, 2.31 mmol, 1.0 equiv) in DMF (5 mL) were added K 2 CO 3 (960 mg, 6.93 mmol, 3.0 equiv) and 5-hydroxypicolinonitrile (612.35 mg, 5.09 mmol, 2.2 equiv) at room temperature. Next, the reaction mixture was stirred at 80 °C for 3 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with ice-cold water (50 mL) and extracted with ethyl acetate (3 × 200 mL). The combined organic layers were washed with water (5 × 50 mL), followed by 1N NaOH solution (3 × 30 mL), and then brine (50 mL), and dried over anhydrous sodium sulfate. Removal of the solvent under reduced pressure gave a crude material, which was purified by combi-flash on silica gel using an ethyl acetate - hexane system as the eluent to give 5,5'-(butane-1,4-diylbis(oxy))dipicolinonitrile (250 mg, 36.7%).

[0228] Step 2 A suspension of ammonium chloride (334.5 mg, 6.25 mmol, 8.0 equiv) in toluene (10 mL) was added dropwise with trimethylaluminum (2 M) (901.5 mg, 3.13 mL, 6.25 mmol, 8.0 equiv) at 0 °C. The mixture was stirred at the same temperature for 10 minutes and then at room temperature for 15 minutes. To this mixture, 5,5’-(butane-1,4-diylbis(oxy))dipicolylnitrile (230 mg, 0.78 mmol, 1.0 equiv) was added and the reaction mixture was stirred at room temperature for an additional 15 minutes. Next, the reaction mixture was stirred under reflux for 16 hours. The reaction mixture was cooled to room temperature, diluted with methanol (5 mL), and stirred at room temperature for 30 minutes. The reaction mixture was diluted with 1N aqueous HCl (20 mL) and washed with ethyl acetate (50 mL). The aqueous layer was basified with 5N NaOH (15 mL) and extracted with a 1:5 mixture of ethanol-ethyl acetate (5 × 80 mL). The combined organic layers were dried over anhydrous sodium sulfate. Removal of the solvent gave a crude material, which was purified by reverse-phase HPLC to give 5,5’-(butane-1,4-diylbis(oxy))dipicolylimidamide as the free base. The solid was dissolved in 1.25 M HCl in EtOH (5 mL), the solution was concentrated under vacuum, and lyophilized to give 5,5’-(butane-1,4-diylbis(oxy))dipicolylimidamide dihydrochloride (20 mg, 7.8%). Analysis data LCMS: 329 [M+1] + 1 HNMR (400 MHz, DMSO-d 6 ) δ 9.40 (brs, 4H), 9.15 (brs, 1H), 8.49 (d, 2H), 8.30 (dd, 2H), 7.75 (dd, 2H), 4.26 (brs, 4H), 1.90 (brs, 4H).

[0229] Example 17: Preparation of 5-(3-(4-carbamimidoylphenoxy)propoxy)picolinimidamide

Chemical formula

[0230] Step 1 To a stirred solution of benzyl alcohol (5.35 g, 49.5 mmol, 1.2 equiv) in DMSO (30 mL) at 0 °C was added sodium hydride (1.28 g, 53.5 mmol, 1.3 equiv) portionwise, and the resulting mixture was stirred at the same temperature for 15 minutes. To this mixture was added 4-fluorobenzonitrile (5 g, 41.2 mmol, 1 equiv), and then the reaction mixture was stirred at room temperature for 2 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was poured into water (200 mL), the precipitate was filtered off and dried under vacuum to give 4-(benzyloxy)benzonitrile (6.97 g, 80%), which was used in the next step without further purification.

[0231] Step 2 To a stirred solution of 4-(benzyloxy)benzonitrile (6.9 g, 32.9 mmol, 1 equiv) in a solution of MeOH and dioxane (1:1, 80 mL) at 0 °C was added thionyl chloride (39.23 g, 329.7 mmol, 10 equiv) dropwise, and the reaction mixture was stirred at room temperature overnight. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with diethyl ether (500 mL) and stirred for 15 minutes. The precipitate was filtered off and dried under vacuum to give methyl 4-(benzyloxy)benzimidate hydrochloride (5 g, 62%), which was used in the next step without further purification.

[0232] Step 3 To a stirred solution of methyl 4-(benzyloxy)benzimidate hydrochloride (5 g, 20.7 mmol, 1 equiv) in methanol (100 mL) was added 7M ammonia in methanol (50 mL), and the reaction mixture was stirred at 70 °C for 2 hours. The progress of the reaction was monitored by TLC. Next, the methanol was completely evaporated under reduced pressure to give 4-(benzyloxy)benzimidamide (4.4 g, 94%), which was used in the next step without further purification.

[0233] Step 4 To a stirred solution of 4-(benzyloxy)benzimidamide (2 g, 8.8 mmol, 1 equiv) in THF (30 mL) was added a solution of sodium hydroxide (1.05 g, 26.5 mmol, 3 equiv) in water (10 mL), followed by addition of Boc anhydride (5.78 g, 26.5 mmol, 3 equiv). The reaction mixture was stirred at room temperature for 1 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with water (50 ml) and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude material, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as eluent to give tert-butyl (4-(benzyloxy)phenyl)(imino)methylcarbamate (1.5 g, 71%).

[0234] Step 5 To a stirred solution of tert-butyl (4-(benzyloxy)phenyl)(imino)methylcarbamate (1.5 g, 4.6 mmol, 1 equiv) in methanol (100 mL) was added Pd - C (300 mg). The reaction mixture was stirred under a hydrogen atmosphere for 1 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was filtered through a celite pad and washed with methanol (30 mL). The filtrate was concentrated under reduced pressure to give tert-butyl (4-hydroxyphenyl)(imino)methylcarbamate (1.3 g, 86%), which was used in the next step without further purification.

[0235] Step 6 A stirred solution of tert-butyl (4-hydroxyphenyl)(imino)methylcarbamate (0.500 g, 2.1 mmol, 1 equiv) and 1,3-dibromopropane (1.28 g, 6.3 mmol, 3 equiv) in acetone (15 mL) was added potassium carbonate (0.434 g, 3.1 mmol, 1.5 equiv), and the reaction mixture was stirred at 60 °C for 2 h. The progress of the reaction was monitored by TLC. After completion, the solid was removed by filtration, and the filtrate was concentrated to give an oily crude material, which was purified by column chromatography on silica gel to give tert-butyl (4-(3-bromopropoxy)phenyl)(imino)methylcarbamate (340 mg, 45%).

[0236] Step 7 A stirred solution of (Z)-tert-butyl (5-hydroxypyridin-2-yl)methanediylidenedicarbamate (0.280 mg, 0.8 mmol, 1 equiv) and tert-butyl (4-(3-bromopropoxy)phenyl)(imino)methylcarbamate (325 mg, 0.9 mmol, 1.1 equiv) in DMF (10 mL) was added potassium carbonate (0.331 mg, 2.4 mmol, 3 equiv), and the reaction mixture was stirred at 60 °C for 2 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with water (5 × 50 mL), followed by brine (20 mL), and dried over anhydrous sodium sulfate. Removal of the solvent under reduced pressure gave a crude material, which was purified by combiflash on silica gel to give tribock-5-(3-(4-carbamimidoylphenoxy)propoxy)picolinimidamide (350 mg, 69%).

[0237] Step 8 A solution of tribock-5-(3-(4-carbamimidoylphenoxy)propoxy)picolinimidamide (0.350 mg, 0.5 mmol, 1 equiv) in 4 M solution of HCl in dioxane was stirred at room temperature for 5 h. 1The progress of the reaction was monitored by \(^1\)H NMR. After completion, the reaction mixture was triturated with ethyl acetate, the filtrate was separated, and dried under vacuum to obtain 5-(3-(4-carbamimidoylphenoxy)propoxy)picolinimidamide dihydrochloride (152 mg, 85%). Analysis data LCMS: 313.15 [M+1] + 1 \(^1\)H NMR (400 MHz, DMSO-d 6 ) δ ppm 9.40 (brs., 2H) 9.15 (brs, 2H) 9.20 (brs, 2H) 8.91 (brs, 2H) 8.51 (d, 1H) 8.33 (d, 1H) 7.84 (d, 2H) 7.76 (dd, 1H) 7.18 (m, 2H) 4.37 (t, 2H) 4.28 (t, 2H) 2.24 - 2.31 (m, 2H).

[0238] Example 18: Preparation of 5-{2-[(1R,3S)-3-[2-(4-carbamimidoylphenyl)ethyl]cyclohexyl]ethyl}pyridine-2-carboximidamide

Chemical formula

[0239] Step 1 To a solution of 5-methylpicolinonitrile (5 g, 42.30 mmol, 1.0 eq) in CHCl 3 (80 mL) was added AIBN (3.47 g, 21.15 mmol, 0.5 eq) at room temperature, followed by NBS (15.05 g, 84.60 mmol, 2.0 eq), and the mixture was stirred at 50 °C for 3 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with water (150 mL) and extracted with dichloromethane (3 × 300 mL). The combined organic layers were washed with brine and dried over anhydrous sodium sulfate. Removal of the solvent under reduced pressure gave a crude product, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to obtain 5-(bromomethyl)picolinonitrile (2.5 g, 30%) as a brown solid.

[0240] Project 2 A mixture of 5-(bromomethyl)picolinonitrile (2.5 g, 12.69 mmol, 1.0 eq) and triethyl phosphite (2.7 mL, 15.22 mmol, 1.2 eq) was stirred at 140 °C for 4 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with ice-cold water (150 ml) and extracted with ethyl acetate (3 × 200 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous sodium sulfate. Removal of the solvent under reduced pressure gave a crude product, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to give diethyl (6-cyanopyridin-3-yl)methylphosphonate (2.5 g, 83%) as a viscous liquid.

[0241] Project 3 To a solution of diethyl (6-cyanopyridin-3-yl)methylphosphonate (0.316 g, 1.24 mmol, 1.5 eq) in THF (10 mL) at 0 °C was added dropwise a 1 M solution of potassium tert-butoxide in THF (1.24 mL, 1.24 mmol, 1.5 eq), and the reaction mixture was stirred at the same temperature for 15 min. To this solution was added a solution of 4-(2-((1S,3S)-3-formylcyclohexyl)ethyl)benzonitrile (0.2 g, 0.828 mmol, 1 eq) in THF (5 mL), and the reaction mixture was stirred at room temperature for 45 min. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with an aqueous ammonium chloride solution (40 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with brine (20 mL) and dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude material, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to give 5-((E)-2-((1S,3S)-3-(4-cyanophenethyl)cyclohexyl)vinyl)picolinonitrile (0.180 g, 63.8%).

[0242] Project 4 To a solution of 5-((E)-2-((1S,3S)-3-(4-cyanophenethyl)cyclohexyl)vinyl)picolinonitrile (0.130 g, 0.380 mmol, 1 equiv) in methanol (20 mL) was added Pd-C (7 mg). The reaction mixture was stirred at room temperature for 25 minutes under a hydrogen atmosphere. The progress of the reaction was monitored by TLC and 1 1H NMR. After completion, the reaction mixture was filtered through a Celite pad, and the pad was washed with methanol (20 mL). The filtrate was concentrated under reduced pressure to give 5-(2-((1R,3S)-3-(4-cyanophenethyl)cyclohexyl)ethyl)picolinonitrile (100 mg), which was used in the next step without further purification.

[0243] Step 5 To a suspension of NH 4 Cl (161 mg, 3.02 mmol, 8 equiv) in toluene (5 mL) at 0 °C was added dropwise 2M trimethylaluminum in toluene (1.5 mL, 3.02 mmol, 8 equiv), and the mixture was stirred at the same temperature for 15 minutes. The mixture was brought to room temperature and stirred for an additional 10 minutes. To this mixture was added a solution of 5-(2-((1R,3S)-3-(4-cyanophenethyl)cyclohexyl)ethyl)picolinonitrile (130 mg, 0.378 mmol, 1 equiv) dissolved in toluene (5 mL), and the reaction mixture was stirred at room temperature for an additional 10 minutes and then at 120 °C for 18 hours. The reaction mixture was cooled to room temperature, diluted with methanol (5 mL), and stirred at room temperature for 15 minutes. This reaction mixture was diluted with 3M aqueous HCl (15 mL) and washed with ethyl acetate (30 mL). The aqueous layer was basified with 3M aqueous NaOH solution. NaOH solution was added, and the mixture was extracted with 20% ethanol-ethyl acetate solution (3 × 50 mL). The combined organic layers were dried over sodium sulfate and concentrated under vacuum to give a crude material, which was purified by reverse-phase HPLC to give the desired product as the free base. The solid was dissolved in 1.25M HCl in ethanol (3 mL), concentrated to give a solid, which was lyophilized to give the desired compound as the dihydrochloride (20 mg, 11.7%). Analytical data LCMS: 378.3 [M+1] + 1 H NMR (400 MHz, CD 3 OD) δ 8.68 (d, 1H) 8.10 (d, 1H) 7.94 (dd, 1H) 7.73 (d, 2H) 7.45 (d, 2H) 2.85 - 2.70 (m, 4H) 1.95 - 1.75 (m, 4H), 1.70 - 1.50 (m, 4H), 1.40 - 1.20 (m, 4H) 1.00 - 0.77 (m, 2H).

[0244] Example 19: Preparation of 4-{[5-(4-carbamimidoylphenoxy)pentyl]oxy}pyridine-2-carboximidamide

Chemical formula

[0245] Step 1 To a stirred solution of pentane-1,5-diol (4.5 g, 43.47 mmol, 3.0 equiv) in DMF (15 mL) at 0 °C was added NaH (360 mg, 21.6 mmol, 1.5 equiv), and the resulting mixture was stirred at 0 °C for 15 minutes. To this mixture was added 4-chloropyridine-2-carbonitrile (2 g, 14.4 mmol, 1.0 equiv), and the reaction mixture was stirred at room temperature for 12 hours. The progress of the reaction was monitored by TLC. After 4-chloropyridine-2-carbonitrile was consumed, the reaction mixture was diluted with ethyl acetate (100 mL) and washed with water (3 × 50 mL). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure to give a crude oil, which was purified by combiflash on silica gel using an ethyl acetate - hexane system as the eluent to give 4-[(5-hydroxypentyl)oxy]pyridine-2-carbonitrile (1 g, 34.38%). Analysis data LCMS: 206 ([M + 1]) +

[0246] Step 2 To a stirred solution of 4-[(5-hydroxypentyl)oxy]pyridine-2-carbonitrile (500 mg, 2.42 mmol, 1.0 equiv) in DMF (10 mL) at 0 °C was added NaH (291 mg, 7.27 mmol, 1.5 equiv), and the resulting mixture was stirred at 0 °C for 15 min. To this mixture was added 4-fluorobenzonitrile (588 mg, 4.85 mmol, 2.0 equiv), and the reaction mixture was stirred at room temperature for 12 h. The progress of the reaction was monitored by TLC. After 4-[(5-hydroxypentyl)oxy]pyridine-2-carbonitrile was consumed, the reaction mixture was diluted with ethyl acetate (100 mL) and washed with water (3 × 20 mL). The organic layer was dried over sodium sulfate. Removal of ethyl acetate under reduced pressure gave a crude oil, which was purified by combiflash on silica gel using an ethyl acetate–hexane system as eluent to afford 4-{[5-(4-cyanophenoxy)pentyl]oxy}pyridine-2-carbonitrile (300 mg, 40.26%).

[0247] Step 3 NH in toluene (10 mL) 4To a stirred suspension of Cl (418 mg, 7.81 mmol, 8 eq) at 0 °C, trimethylaluminum (4.0 mL, 7.81 mmol, 8 eq) was added dropwise under nitrogen. The reaction mixture was stirred at 0 °C for 10 minutes and then at room temperature for 15 minutes. The reaction mixture was cooled to 0 °C and 4-{[5-(4-cyanophenoxy)pentyl]oxy}pyridine-2-carbonitrile (300 mg, 0.97 mmol, 1 eq) was added. The reaction mixture was stirred at room temperature (RT) for 15 minutes and then at 120 °C for 16 hours. The reaction mixture was cooled to 0 °C, methanol (5 mL) was added dropwise, and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was acidified with 2M HCl solution (150 mL) and extracted with ethyl acetate (50 mL). The organic layer was separated, the aqueous layer was basified using 5N NaOH solution (50 mL), and extracted with 20% ethanol-ethyl acetate (5 × 200 mL). The combined organic layers were dried over sodium sulfate, filtered, and evaporated to dryness. A trace amount of water was removed with a toluene azeotrope to obtain a solid residue. The solid was triturated with 1:1 ethanol-ethyl acetate (2 × 200 mL) and filtered. After the filtrate was evaporated to dryness, the residue was purified by reverse-phase HPLC to obtain 4-{[5-(4-carbamimidoylphenoxy)pentyl]oxy}pyridine-2-carboximidamide as the free base. This solid was dissolved in 1.25M HCl in ethanol (5 mL) at 0 °C, the material was evaporated to dryness, and then lyophilized to obtain 4-{[5-(4-carbamimidoylphenoxy)pentyl]oxy}pyridine-2-carboximidamide as the hydrochloride salt (10 mg, 2.5%). Analysis data LCMS: 342 [M+1] + 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.58 (brs, 2H), 9.38 (brs, 2H), 9.21 (brs, 2H), 8.95 (brs, 2H), 8.60 (d, 1H), 7.99 (s, 1H), 7.84 (d, 2H), 7.34 (d, 1H), 7.15 (d, 2H), 4.23 (t, 2H), 4.12 (t, 2H), 1.90 - 1.70 (m, 4H), 1.65 - 1.50 (m, 2H).

[0248] Example 20: Cytotoxicity of Pyridinyl Analogue Compounds The purpose of this study was to investigate the potential cytotoxic effects of Compounds 1 - 15 against three cancer cell lines. The 50% inhibitory concentration (IC 50 ) of these compounds in various cancer cell lines was determined using the CellTiter - Glo™ Luminescent Cell Viability Assay at different compound concentrations. Each cell line (e.g., NCI - H209; NCI - H69; and SW1271) was treated with Compounds 1 - 15, and the medium contained a 0.2% [v / v] DMSO vehicle control. All cells were cultured in medium supplemented with 10 - 20% fetal bovine serum at 37°C, 5% CO 2 , and 95% humidity. The IC 50 values for the NCI - H209; NCI - H69; and SW1271 cell lines are shown in Table 2 below. [Table 2]

[0249] Example 21. Cytotoxicity of Pyridinyl Compounds The purpose of this study was to investigate the effect of Compound 1 on cytotoxicity against 61 different types of cancer cell lines. The IC 50 values were obtained using CellTiter - Glo™ as described above. Each cell line shown in Table 3 was treated with Compound 1 and cisplatin, a standard chemotherapeutic agent as a reference control, and the medium contained a 0.2% [v / v] DMSO vehicle control. All cancer cell lines were cultured in medium supplemented with 10 - 20% fetal bovine serum at 37°C, 5% CO 2 , and 95% humidity. Table 3 shows the IC 50 values of Compound 1 against various types of liver cancer, cholangiocarcinoma, gallbladder cancer, renal cancer, prostate cancer, lung cancer, brain cancer, ovarian cancer, gastric cancer, colon cancer, and bone cancer. [Table 3] TIFF2025081626000144.tif135118

[0250] Hepatocyte cell line The purpose of this study was to investigate the effect of Compound 1 on the cell viability of various types of liver cancer. In this study, 10 different hepatocellular carcinoma cell lines were used. Each hepatocyte cell line shown in Table 4 was treated with Compound 1, pentamidine, and standard treatment (cisplatin) as a reference control, and the medium contained a 0.2% [v / v] DMSO vehicle control. The IC 50 values were obtained as described above. The raw data values from the CellTiter-Glo™ cell viability assay, expressed in relative light units, were normalized to the vehicle for each individual plate, and the decrease in luminescence indicated a decrease in viability (%). The data were analyzed using GraphPad PRISM with a non-linear sigmoid plot using a variable slope (asymmetric 4-point linear regression), and the IC 50 values for each compound were generated. In the experiment, pentamidine, Compound 1, and the cisplatin standard treatment control were tested in complete growth medium for 8 days. The cells were first treated with the test compounds on day 0, and then the cells were replenished with fresh compound dilutions on day 3. Pentamidine, Compound 1, and cisplatin were tested at 9 concentration points: 100, 33.33, 11.11, 3.70, 1.23, 0.41, 0.14, 0.05, and 0.02 μM (final DMSO concentration = 0.5%). One independent experiment was performed, and the IC 50 values are summarized in Table 4 below. In particular, the IC 50 values for pentamidine and Compound 1 were 0.8 and 0.6 μM, respectively, in Hep3B-luc. The IC 50 value for cisplatin was 5.1 μM, which was consistent with past data.

Table 4

[0251] Furthermore, the IC 50 values of Compounds 4, 5, and 19 were also evaluated in a similar experiment and showed strong cytotoxicity against hepatocyte cell lines as shown in Table 5.

Table 5

Table 6

[0252] Example 22. Orthotopic In Vivo Analysis In this study, the in vivo therapeutic effect of Compound 1 against liver cancer was evaluated in an orthotopic mouse model using the Hep3B2.1-7-Luc cell line. Three groups of an orthotopic model of BALB / c nude mice were treated orally (po) Q3D for 1 week followed by QD for 3 weeks with vehicle, 10 mg / kg of Compound 1, and 20 mg / kg of Compound 1. The results of the total flux measured in units of photons / second / 10 6 are shown in Figure 1. The average total flux (photons / second / 10 6 ) of the mice in the vehicle control group reached 1223.01 photons / second / 10 6 on day 28 after randomization. The average total flux (photons / second / 10 6 ) on day 28 after randomization was 464.74 and 306.74 in the Compound 1 (10 mg / kg) group and the Compound 1 (20 mg / kg) group, respectively. Compared with the vehicle group, the Compound 1 (10 mg / kg) group showed a significant antitumor effect from day 8 to day 18 and day 28, and the Compound 1 (20 mg / kg) group showed a significant antitumor effect from day 8 to day 28 (Figure 1). Compound 1 was highly resistant in both the 10 mg / kg group and the 20 mg / kg group, and there was no obvious weight loss. Both Compound 1 groups steadily increased their body weight until day 28, while the mice in the vehicle-treated group showed weight loss on day 28. The weights of the whole liver (including the tumor) are shown in Table 7 and Figure 2. [Table 7]

[0253] The tumor size (indicated by signal intensity) of the Compound 1 (10 mg / kg) group showed a significant difference on day 25 and a highly significant difference on day 28. The tumor size of Compound 1 (20 mg / kg) showed a highly significant difference from day 22 to day 28 compared with the vehicle control group based on two-way ANOVA and Bonferroni post-test.

[0254] The tumor sizes in the compound 1 (10 mg / kg) group did not show significant differences during the treatment days. The tumor sizes of compound 1 (20 mg / kg) showed significant differences on day 28 and very significant differences on day 25 compared with the vehicle control group based on statistical analysis using non-parametric analysis of variance followed by the Kruskal Wallis test.

[0255] One-way analysis of variance and Dunnett's post-test were combined to compare the results of serum-derived blood chemistry (including ALT, AST, ALP, TP, ALB, UA, UREA, Glu, TC, TG, Ca, Mg, P, CK, LDH, GLB, A / G, and CREA) between the vehicle and treatment groups. Non-parametric analysis of variance followed by the Kruskal Wallis test was performed to compare serum-derived blood chemistry (including ALT, AST, and ALP) between the vehicle group and the treatment groups. Both treatment groups (10 mg / kg and 20 mg / kg) showed significant decreases in the levels of liver injury / damage markers, namely ALT and AST, in a dose-dependent manner, suggesting that the treatment with compound 1 enhanced liver function by reducing the burden of liver tumors (Figure 4).

[0256] In summary, compound 1 was highly resistant at both dose levels tested. Furthermore, compound 1 administered at 20 mg / kg showed significant in vivo antitumor activity against the Hep3B2.1-7-Luc liver orthotopic model in BALB / c nude mice on days 25 and 28 (Figures 1-4).

[0257] Example 23. Resistance of Compound 1 in BALB / c Nude Mice The purpose of this study was to evaluate the resistance of compound 1 in non-tumor-bearing BALB / c nude mice. The exposure of compound 1 in plasma, liver, kidney, colon, bladder, and serum-derived blood chemistry was tested.

[0258] Materials Thirty-six 6- to 8-week-old female BALB / c nude mice weighing approximately 19 to 23 g were housed in individual ventilated cages with three animals per cage at a constant temperature (20 to 26 °C) and humidity (40 to 70%). The animals had free access to irradiated sterilized dry pellet feed and sterilized drinking water throughout the study period. The details of the research plan are shown in Table 8.

Table 8

[0259] After grouping, the animals were checked daily for morbidity and mortality. At the time of regular monitoring, the animals were measured for any effects on behavior, such as movement, food and water consumption, and weight gain / loss. Body weight was measured daily. Deaths and other clinical signs were recorded.

[0260] Animals in each group were tested and samples were collected at two different time points for blood chemistry tests and compound concentrations in plasma, kidney, liver, colon, and bladder. The first sampling was performed immediately before the last dose (0 h) on day 21. The second sampling was conducted 1 h after the last dose on day 21 (1 h). The detailed description of the sampling method is as follows.

[0261] Serum collection: Approximately 500 μL of blood was collected into 1.5 mL tubes. Before centrifuging all samples, they were placed at room temperature for 30 min, and then the blood was centrifuged at 6,000 rpm at 4 °C for 5 min to obtain serum. Serum samples were transferred to an -80 °C freezer for storage for blood routine tests.

[0262] Plasma collection: Approximately 200 μL of blood was collected into 1.5 mL tubes containing the plasma anticoagulant 2K-EDTA. Plasma was transferred to an -80 °C freezer for storage for exposure analysis.

[0263] Kidney collection: The left kidney of each mouse was collected, weighed, snap-frozen in dry ice, and transferred to an 80 °C freezer for storage for exposure analysis. For paraffin embedding, H&E staining, and image analysis, the right kidney was fixed in neutral formalin for 24 hours and then in 70% EtOH.

[0264] Liver collection: The left liver lobe of each mouse was collected and divided into two parts. One part was weighed, snap-frozen in dry ice, and transferred to an 80 °C freezer for storage for subsequent exposure analysis. For paraffin embedding, H&E staining, and image analysis, the other part was fixed in neutral formalin for 24 hours and then in 70% EtOH.

[0265] Colon collection: The entire colon was collected, and then the entire colon was manually perfused with cold PBS solution to remove fecal matter. Finally, the colon was opened horizontally and gently blotted. The entire colon of each mouse was collected, weighed, snap-frozen in dry ice, and then transferred to an -80 °C freezer for storage. The entire colon of Group 5 (Compound 1, 20 mg / kg, oral administration, QD*21 group) was stored for exposure analysis.

[0266] Bladder collection: The bladder of each mouse was collected, weighed, snap-frozen in dry ice, and then transferred to an -80 °C freezer for storage. The bladders of Group 5 (Compound 1, 20 mg / kg, oral administration, QD*21 group) were stored for exposure analysis and subjected to bioassay analysis. The others were undetermined.

[0267] Results The body weight changes of mice and the percentage of relative change in body weight (RCBW) are shown in Figures 5 and 6. Overall, in the 5 mg / kg QD, 10 mg / kg QD, 10 mg / kg Q2D, and 20 mg / kg QD groups, Compound 1 had high tolerance and no obvious weight loss was observed. However, in the Compound 1 (40 mg / kg QD) group, compared with the vehicle control group, the body weight was statistically significantly lower on the 20th and 21st days (Figures 5 and 6). No mice died in all experimental groups during the study period.

[0268] Example 24. Pharmacokinetics of Compound 1 and Compound 5 in the Liver The PK of compound 1 (20 mg / kg), compound 5 (10 mg / kg), and pentamidine (20 mg / kg) was tested. Tissue samples were collected as described in Example 23 above. Briefly, blood samples (about 50 - 60 μL) were collected from the posterior orbital plexus of mice under light isoflurane anesthesia at 0.5, 1, 3, 8, 48, and 72 hours. Plasma samples were separated by centrifugation at 2,000 xg for 6 minutes and stored at less than -70 ± 10 °C until bioanalysis. Immediately after blood collection, excess CO 2 Asphyxiation was used to euthanize the animals, and samples were collected from a set of 5 mice at each time point. The collected tissue samples were immediately rinsed 3 times in ice-cold PBS (5 - 10 seconds / rinse, using 5 - 10 mL of fresh PBS in a disposable Petri dish for each rinse) and dried with absorbent paper. The tissue samples were homogenized using ice-cold phosphate-buffered saline (pH 7.4), and the homogenate was stored at less than -70 ± 10 °C until analysis. The total volume of the homogenate was 3 times the tissue weight excluding the liver sample, and the total volume of the homogenate was 10 times the liver weight. The bioanalysis process was determined by an LC-MS / MS method suitable for the purpose.

[0269] Calibration standards were prepared by spiking the test compound into blank plasma. 10 μL of the working calibration standard was spiked into 190 μL of blank mouse plasma or tissue homogenate to generate linearly spiked calibration standards. The calibrator concentrations were 5,000, 2,000, 1,000, 200, 100, 20, 10, 2, and 1 ng / mL. The calibration standard samples were processed together with the test samples. A 25 μL aliquot of the plasma or tissue homogenate test sample was treated with 100 μL of acetonitrile containing an internal standard (500 ng / mL glypidide). The sample was vortexed for 5 minutes. The sample was centrifuged at 4000 rpm for 10 minutes at 4 °C. After centrifugation, 100 μL of the clear supernatant was transferred to a 96-well plate and analyzed using LC-MS / MS. Chromatographic separation was achieved using a Kintex Polar column (C18, 100X4.6 mm, 5 μ) and a column oven temperature of 45 °C. The mobile phase A for compound 1 diHCl was water containing 0.1% formic acid in acetonitrile, and the mobile phase B was 10 mM ammonium formate. The mobile phase A for the pentamidine compound and compound 5 was water with 0.1% formic acid, and the mobile phase B was acetonitrile with 0.1% formic acid.

[0270] The gradient program for Compound 1 was 5% B (1 - 2.40 minutes), 90% B (2.60 - 3.00 minutes), with the initial value of B being 90%. The retention rate was 1.47 / minute, and the internal standard glypidide was 1 / 9 minute. The column was maintained at 45 °C. For pentamidine and Compound 5, the analysis was performed using 233 MassSpec with a Turbo Ion Spray interface operating in positive ionization mode and an AB Sciex API5000. Quantification was carried out using the multiple reaction monitoring (MRM) method, with the transition of (m / z) 328a (a is grave) (m / z) 311 for Compound 5 and (m / z) 548a (a is grave) (m / z) 366 for edoxaban (internal standard). On the other hand, for pentamidine, it was (m / z) 341a (a is grave) (m / z) 324, and for verapamil (internal standard), it was (m / z) 548a (a is grave) (m / z) 366.

[0271] The gradient program for Compound 5 was 10% B (0 - 0.2 minutes), 95% B (1.5 - 2 minutes), with the initial value of B being 10% and stopping at 2.6 minutes. The flow rate was 0.5 mL / minute. The gradient program for pentamidine was 10% B (0 - 0.2 minutes), 95% B (1.4 - 2 minutes), with the initial value of B being 10% and stopping at 2.5 minutes. The flow rate was 0.5 mL / minute.

[0272] As shown in Figure 7, Compound 1 (20 mpk) and Compound 5 (10 mpk) showed greater exposure to the liver than pentamidine (20 mpk) after oral administration (p.o.). When C max was normalized for the dose of Compound 1 (20 mpk, p.o.), it was approximately 7 times higher than that of pentamidine (20 mpk, p.o.). Compound 5 administered at 10 mpk, p.o. showed an exposure rate similar to that of Compound 1 administered at 20 mpk, p.o. Both Compound 1 and 5 showed a longer half-life than pentamidine. Compound 5 showed a longer half-life (about 3 times) than Compound 1. Figure 8 shows the exposure rate of Compound 1 in the liver / kidney / small intestine / ileum / plasma.

[0273] In summary, compounds 1 and 5 showed increased exposure in the liver, small intestine, and ileum, but had lower exposure to plasma compared to other tissues.

[0274] Example 25. In Vitro Evaluation of the Cytotoxicity of Compound 1 In the experiment, pentamidine, compound 1, and the standard treatment control cisplatin were tested in complete growth medium for 8 days. Hep-3b cells were first treated with the test compounds on day 0, and then the cells were replenished with fresh compound dilutions on day 3. Compounds 1, pentamidine, and cisplatin were tested at nine concentration points: 100 μM, 33.33 μM, 11.11 μM, 3.70 μM, 1.23 μM, 0.41 μM, 0.14 μM, 0.05 μM, and 0.02 μM (final DMSO concentration = 0.5%). The raw data values from the CellTiter-Glo™ cell viability assay, expressed in relative light units, were normalized to the vehicle for each individual plate, and the decrease in luminescence indicated a decrease in viability (%). The data were analyzed in GraphPad PRISM using a non-linear sigmoid plot with a variable slope (asymmetric four-parameter linear regression), and the IC 50 values for each compound were generated. The dose-response curves are shown in Figure 7. The IC 50 values were generated based on the normalized dose-response curves. The IC 50 values for pentamidine and compound 1 were 0.8 and 0.6 μM, respectively, in Hep-3b lux. The IC 50 value for cisplatin (5.1 μM) was consistent with past data.

[0275] Example 26: Orthotopic Colorectal In Vivo Study This example shows the resistance and effect of compound 1 on tumor growth in an orthotopic colorectal cancer study.

[0276] Method Approximately 2.0 × luciferase stably expressing colorectal cancer tumor cells (COLO205-Luc) cells suspended in 30 μl of DPBS were injected into the cecal wall of BALB / c nude mice.

[0277] Animals were selected to be grouped when the bioluminescence intensity increased in three consecutive measurements on the 20th day after tumor transplantation. This indicated that the tumor was in the growth phase (the average bioluminescence measurement reached 2.13x10 7 photons / sec). Animals were assigned to groups using Excel-based randomization software that performed stratified randomization based on bioluminescence intensity. Treatment was initiated according to a predetermined regimen as shown in the experimental schedule.

[0278] Mice were administered Compound 1 at 10 mg / k per day, 20 mg / kg per day, 10 mg / kg twice a day, 20 mg / kg twice a day, or 40 mg / kg per day.

Table 9

[0279] The primary efficacy endpoint was bioluminescence (intensity value and change from baseline). The body weights of surgically inoculated mice were measured, and luciferin was administered intraperitoneally at a dose of 150 mg / kg. Ten minutes after the luciferin injection, the animals were pre-anesthetized with a mixed gas of oxygen and isoflurane. Once the animals were fully anesthetized, they were transferred to the imaging chamber for bioluminescence measurement using an IVIS (Lumina II) imaging system. The whole-body bioluminescence of animals including primary and metastatic tumors was measured and recorded once a week.

[0280] Tumor growth inhibition (TGI) was calculated for each group using the formula: TGI (%) = [1 - (Ti - T0) / (Vi - V0)] × 100. Ti represents the average tumor bioluminescence value of the treatment group on a specific day, T0 represents the average tumor bioluminescence value of the treatment group on day 0, Vi represents the average tumor bioluminescence value of the vehicle control group on the same day as Ti, and V0 represents the average tumor bioluminescence value of the vehicle group on day 0.

[0281] Tumor weight was measured at the end of the study. T / C 重量 value (percent) was calculated using the formula: T / C 重量 % = T 重量 / C 重量Calculated using × 100%, where T 重量 and C 重量 were the mean tumor weights of the treatment group and the vehicle control group, respectively.

[0282] To compare between three or more groups, one-way analysis of variance was performed. When a non-significant F statistic (the ratio of treatment variance to error variance) was obtained, the comparison between groups was performed using Dunnett's (two-sided). All data were analyzed using SPSS 17.0. p < 0.05 was considered statistically significant.

[0283] Results The body weights of the animals were regularly monitored as an indirect measure of toxicity. There was no group in which body weight clearly decreased as a result of the administration of Compound 1 (Table 10 and Figure 10). Figure 10 shows the relative change in body weight (%) from the first day of administration. The data points represent the mean change rate of body weight for the group. The error bars represent the standard error of the mean (SEM). There were no deaths or morbidity. Therefore, there is no obvious toxicity associated with the administration of Compound 1 to BALB / c nude mice bearing tumors. [Table 10]

[0284] Tumor size in response to treatment with Compound 1 was measured using bioluminescence. [Table 11]

[0285] Figure 11 is a plot of relative bioluminescence over time as a measure of tumor growth.

[0286] This study indicates that Compound 1 has no significant toxicity and is highly resistant. Furthermore, a non-statistically significant trend was observed indicating that Compound 1 administered at a dose of 20 mg / kg or 40 mg / kg daily may be effective in reducing tumor growth. Additional studies can be conducted to evaluate a daily dosing schedule of 40 mg / kg according to Example 27.

[0287] Example 27: Follow-up of a Colon Orthotopic In Vivo Study at a 1-Day Dose of 40 mg / kg Methods Approximately 2.0 x 10 6 Colo205-luc2 cells suspended in 30 μl of DPBS are injected into the cecal wall of BALB / c nude mice.

[0288] Animals are selected on day 20 after tumor transplantation for grouping when the bioluminescence intensity increases in three consecutive measurements. This indicates that the tumor was in the growth phase (the average bioluminescence measurement reached 2.13x10 7 photons / sec). Animals are assigned to groups using Excel-based randomization software that performs stratified randomization based on bioluminescence intensity. Compound 1 is administered to the mice at 40 mg / kg daily.

[0289] The primary efficacy endpoint is bioluminescence (intensity value and change from baseline). The body weight of surgically inoculated mice is measured, and luciferin is administered intraperitoneally at a dose of 150 mg / kg. Ten minutes after the luciferin injection, the animals are pre-anesthetized with a mixed gas of oxygen and isoflurane. Once the animals are fully anesthetized, they are transferred to the imaging chamber for bioluminescence measurement using an IVIS (Lumina II) imaging system. The whole-body bioluminescence of animals, including primary and metastatic tumors, is measured and recorded once a week.

[0290] Tumor growth inhibition (TGI) is calculated for each group using the formula: TGI (%) = [1 - (Ti - T0) / (Vi - V0)] × 100. Ti represents the average tumor bioluminescence value of the treatment group on a specific day, T0 represents the average tumor bioluminescence value of the treatment group on day 0, Vi represents the average tumor bioluminescence value of the vehicle control group on the same day as Ti, and V0 represents the average tumor bioluminescence value of the vehicle group on day 0.

[0291] The tumor weight is measured at the end of the test. T / C 重量 The value (percentage) is calculated using the formula: T / C 重量 % = T 重量 / C 重量 × 100%, where T 重量 and C 重量 are the average tumor weights of the treatment group and the vehicle control group, respectively.

[0292] To compare between three or more groups, one-way analysis of variance is performed. If a non-significant F statistic (the ratio of treatment variance to error variance) is obtained, the comparison between groups is performed using Dunnett's (two-sided). All data are analyzed using SPSS 18.0.

[0293] Example 28: In Vivo Study of Kidney Homology Balb / c nude mice are injected with 4 x 10^6 luciferases that stably express renal carcinoma tumor cells (ACHN-Luc) in 40 ul of DPBS.

[0294] The whole-body bioluminescence of animals containing primary and metastatic tumors is measured and the images are recorded.

[0295] Before starting the treatment, the body weights of all animals are measured and assigned to two groups using Excel-based randomization software that performs stratified randomization based on body weight. This makes all groups comparable at the baseline. The test animals are administered 40 mg / kg of the compound once a day for 21 days. [Table 12]

[0296] Bioluminescence and the animal's body weight are measured over time.

[0297] Enumerated embodiments

[0298] 1. Formula (I):

Chemical formula

Chemical formula

Chemical formula

Chem.

[0299] 2. Formula (II):

Chem.

Chem.

[0300] 3. Formula (III):

Chem.

Chem.

[0301] 4. Formula (IV):

Chem.

Chem.

[0302] 5.

Chem.

Chem.

Chem.

[0303] 6.

Chem.

Chem.

Chemical formula

[0304] 7.

Chemical formula

Chemical formula

Chem.

[0305] 8.

Chem.

Chem.

Chemical formula

[0306] 9.5-(5-(4-Carbamimidoyl phenoxy) pentyl oxy) picolinimidamide; 6-((5-(4-Carbamimidoyl phenoxy) pentyl) oxy) nicotinimidamide; 5-((5-(4-Carbamimidoyl phenoxy) pentyl) oxy) pyrimidine-2-carboximidamide; 5-((5-(4-Carbamimidoyl phenoxy) pentyl) oxy) pyrazine-2-carboximidamide; 5-(4-(4-Carbamimidoyl phenoxy) butoxy) picolinimidamide; 5-(4-(4-Carbamimidoyl phenoxy) butoxy) picolinimidamide; 5,5’-(Pentane-1,5-diylbis(oxy)) bis(pyrazine-2-carboximidamide); 6,6’-(Heptane-1,7-diyl) dipicolinimidamide; 5,5’-(Heptane-1,7-diyl) dini cotinimidamide; 6,6’-(Heptane-1,7-diyl) dini cotinimidamide; 5-(5-(3-Carbamimidoyl phenoxy) pentyl oxy) picolinimidamide; 4-({5-[(6-Cyanopyridin-3-yl) oxy] pentyl} oxy) pyridine-2-carbonitrile; 5-(((1r,4r)-4-(4-Carbamimidoyl phenoxy) cyclohexyl) oxy) picolinimidamide; 5-(((1S,4S)-4-(4-carbamimidoyl phenoxy) cyclohexyl) oxy) picolinimidamide; 4-(5-(3-carbamimidoyl phenoxy) pentyloxy) picolinimidamide; 5,5’-(butane-1,4-diylbis(oxy)) dipicolinimidamide; 5-(3-(4-carbamimidoyl phenoxy) propoxy) picolinimidamide; 5-{2-[(1R,3S)-3-[2-(4-carbamimidoyl phenyl) ethyl] cyclohexyl] ethyl} pyridine-2-carboximidamide; 4-{[5-(4-carbamimidoyl phenoxy) pentyl] oxy} pyridine-2-carboximidamide; 5-({5-[(6-carbamimidoyl pyridin-3-yl) oxy] pentyl} oxy) pyridine-2-carboximidamide; and A compound selected from the group consisting of 4-({5-[(2-carbamimidoyl pyridin-4-yl) oxy] pentyl} oxy) pyridine-2-carboximidamide.

[0307] 10. A compound having the following structure:

Chemical formula

[0308] 11. A compound having the following structure:

Chemical formula

[0309] 12. A method for treating cancer, comprising administering to a subject suffering from cancer an effective amount of a compound of the following formula (I):

Chemical formula

Chemical formula

Chem.

Chem.

[0310] 13. The method of embodiment 12, wherein m is 1 and n is 1.

[0311] The method of Embodiment 12, where m is 1 and n is 0.

[0312] The method of Embodiment 12, where m is 0 and n is 1.

[0313] The method of Embodiment 12, where m is 1 and n is 2.

[0314] The method of Embodiment 12, where m is 2 and n is 1.

[0315] The method of Embodiment 12, where m is 2 and n is 2.

[0316] The method of Embodiment 12, where m is 0 and n is 0.

[0317] 20. Z 1 or Z 2 is independently selected from the group consisting of N, O, and S, each of which may be substituted, in the method of Embodiment 12.

[0318] 21. Z 1 or Z 2 is independently S, which may be substituted, in the method of Embodiment 12.

[0319] 22. Z 1 or Z 2 is independently O, which may be substituted, in the method of Embodiment 12.

[0320] 23. Z 1 or Z 2 is independently N, which may be substituted, in the method of Embodiment 12.

[0321] 24. Z 1 or Z 2 is independently NR 3 and R 3 is hydrogen, in the method of Embodiment 12.

[0322] 25. Z 1 or Z 2 is independently NR3 and R 3 is selected from the group consisting of alkyl, cycloalkyl, aryl, and heteroaryl, the method of embodiment 12.

[0323] 26. Z 1 or Z 2 is independently NR 3 or CR 5 R 6 is the method of embodiment 12.

[0324] 27. Z 1 is NR 3 and R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, Z 2 is CR 5 R 6 and R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or, R 5 is R 6 together with forms a saturated or partially unsaturated 3- to 9-membered ring, the method of embodiment 12.

[0325] 28. Z 1 or Z 2 is independently CR 5 R 6 and R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, or amino, or, R 5 is R 6 together with forms a saturated or partially unsaturated 3- to 9-membered ring, the method of embodiment 12.

[0326] 29. R 5 or R 6 is independently hydrogen, the method of embodiment 12.

[0327] 30. Z 1 or Z 2 is independently NR 3 and R 3The method of embodiment 12, wherein is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl.

[0328] 31.Z 1 or Z 2 is independently O, S, or SO 2 The method of embodiment 12.

[0329] 32.Y 3 and Y 8 are bonded to the amidine. The method of embodiment 12.

[0330] 33.Y 3 and Y 7 are bonded to the amidine. The method of embodiment 12.

[0331] 34.Y 3 and Y 7 are bonded to the amidine. The method of embodiment 12.

[0332] 35.R 1 and R 2 are independently hydrogen. The method of embodiment 12.

[0333] 36.R 1 is R 2 together with forms a saturated, unsaturated, or partially unsaturated 3- to 9-membered cyclic group (e.g.,

Chemical formula

[0334] 37.R 1 is R 2 together with forms a 5-, 6-, or 7-membered cycloalkyl. The method of embodiment 12.

[0335] 38.R 1 is R 2 together with forms a 6-membered cycloalkyl. The method of embodiment 12.

[0336] 39.R 1 is R2 The method of Embodiment 12 that forms a 7-membered cycloalkyl together.

[0337] 40.Y 1、2、4、5、6、及び8 is CR 7 (e.g., -CH), and Y 2 is N, and Y 3 and Y 7 is bonded to the amidine. The method of Embodiment 12.

[0338] 41.Y 1、4、5、6、及び7 is -CH, and Y 2 is N, and Y 3 and Y 8 is CR 7 and R 7 is amidine. The method of Embodiment 12.

[0339] 42.Y 1、4、5、6、及び8 is -CH, and Y 3 is N, and Y 2 and Y 7 is CR 7 and R 7 is amidine. The method of Embodiment 12.

[0340] 43.Y 1、4、5、6、及び8 is -CH, and Y 3 is N, and Y 2 and Y 7 is CR 7 and R 7 is amidine, m is 1, and n is 0. The method of Embodiment 12.

[0341] 44.Y 1、4、5、及び6 is -CH, and Y 3 and Y 8 is N, and Y 2 and Y 7 is CR 7 and R 7 is amidine, m is 1, and n is 0. The method of Embodiment 12.

[0342] 45. The method of embodiment 12, wherein the cancer is selected from the group consisting of liver cancer, bile duct cancer, osteosarcoma, melanoma, breast cancer, renal cancer, prostate cancer, gastric cancer, colorectal cancer, thyroid cancer, head and neck cancer, ovarian cancer, pancreatic cancer, nerve cancer, lung cancer, uterine cancer, leukemia, and lymphoma.

[0343] 46. The method of embodiment 45, wherein the cancer is liver cancer.

[0344] 47. The method of embodiment 45, wherein the cancer is bile duct cancer.

[0345] 48. The method of embodiment 45, wherein the cancer is prostate cancer.

[0346] 49. The method of embodiment 45, wherein the cancer is pancreatic cancer.

[0347] 50. The method of embodiment 45, wherein the cancer is lung cancer.

[0348] 51. The method of embodiment 45, wherein the cancer is small cell lung cancer.

[0349] 52. The method of embodiment 45, wherein the cancer is non-small cell lung cancer.

[0350] 53. The method of embodiment 45, wherein the cancer is breast cancer.

[0351] 54. The method of embodiment 45, wherein the cancer is colorectal cancer.

[0352] 55. The method of embodiment 45, wherein the cancer is renal cancer.

[0353] 56. The compound of formula (I) or a pharmaceutically acceptable salt thereof is administered to the subject (e.g., a human patient) orally, intravenously or subcutaneously at a dose of about 0.5 mg / kg, 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, about 100 mg / kg, about 110 mg / kg, about 120 mg / kg, about 130 mg / kg, about 140 mg / kg, about 150 mg / kg, about 160 mg / kg, about 170 mg / kg, about 180 mg / kg, about 190 mg / kg, about 200 mg / kg, about 210 mg / kg, about 220 mg / kg, about 230 mg / kg, about 240 mg / kg, about 250 mg / kg, about 260 mg / kg, about 270 mg / kg, about 280 mg / kg, about 290 mg / kg, about 300 mg / kg, about 350 mg / kg, about 400 mg / kg, about 450 mg / kg, about 500 mg / kg, or about 600 mg / kg, according to the method of embodiment 12.

[0354] 57. The method according to embodiment 12, wherein the subject is a human patient.

[0355] 58. The method according to embodiment 12, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered orally to the human patient.

[0356] 59. The method according to embodiment 12, wherein the subject is administered from about 1 mg / kg to about 200 mg / kg per day.

[0357] 60. The method according to embodiment 12, wherein the subject is administered from about 1 mg / kg to about 100 mg / kg per day.

[0358] 61. The method according to embodiment 12, wherein the subject is administered from about 1 mg / kg to about 50 mg / kg per day.

[0359] 62. The method of embodiment 12, wherein the subject is administered from about 0.5 mg / kg to about 50 mg / kg daily.

[0360] 63. The method of embodiment 12, wherein the subject is administered about 2 mg / kg daily.

[0361] 64. A method of treating cancer, comprising administering an effective amount of a compound of formula (V).

Chem.

Chem.

Chem.

[0362] 65.Z 1 or Z 2 is independently selected from the group consisting of O, N, and S, each of which may be substituted, the method of embodiment 64.

[0363] 66.Z 1 or Z 2 is independently O and may be substituted, the method of embodiment 64.

[0364] 67.Z 1 or Z 2 is independently S and may be substituted, the method of embodiment 64.

[0365] 68.Z 1 or Z 2 is independently NR 3 and R 3 is hydrogen, the method of embodiment 64.

[0366] 69.Z 1 or Z 2 is independently NR 3 and R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, the method of embodiment 64.

[0367] 70.Z 1 or Z 2 is independently CR 5 R 6 and is the method of embodiment 64.

[0368] 71.Z 1 is NR 3 and R 3 is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl, and Z 2 is CR 5 R 6 and R 5 or R 6 is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R 5 is R 6The method of Embodiment 64 that together forms a saturated or partially unsaturated 3- to 9-membered ring.

[0369] 72.Y 3 and Y 8 are independently amidine in the method of Embodiment 64.

[0370] 73.Y 3 and Y 7 are independently amidine in the method of Embodiment 64.

[0371] 74.Y 2 and Y 7 are independently amidine in the method of Embodiment 64.

[0372] 75.Y 1、2、4、5、6、及び8 is CR 7 (e.g., -CH), Y 2 is N, Y 3 and Y 7 are bonded to amidine in the method of Embodiment 64.

[0373] 76.Y 1、4、5、6、及び7 is -CH, Y 2 is N, Y 3 and Y 8 is CR 7 and R 7 is amidine in the method of Embodiment 64.

[0374] 77.Y 1、4、5、6、及び8 is -CH, Y 3 is N, Y 2 and Y 7 is CR 7 and R 7 is amidine in the method of Embodiment 64.

[0375] 78.Y 1、4、5、6、及び8 is -CH, Y 3 is N, Y 2 and Y 7 is CR 7 and R 7 is amidine in the method of Embodiment 64.

[0376] 79.Y 1、4、5、及び6 is -CH, and Y 3 3 and Y 8 is N, and Y 2 and Y 7 is CR 7 and R 7 is amidine, the method of embodiment 64.

[0377] 80.R 3 is hydrogen, the method of embodiment 64.

[0378] 81.R 3 is alkyl, the method of embodiment 64.

[0379] 82.R 3 is methyl, the method of embodiment 64.

[0380] 83.R 3 is cycloalkyl, the method of embodiment 64.

[0381] 84.R 3 is aryl, the method of embodiment 64.

[0382] 85.R 3 is heteroaryl, the method of embodiment 64.

[0383] 86.R 5 or R 6 is, independently, hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, amino, or, R 5 is R 6 together with R forms a saturated or partially unsaturated 3 - to 9 - membered ring, the method of embodiment 64.

[0384] 87.R 5 and R 6 are hydrogen, the method of embodiment 64.

[0385] 88.R 7 is independently hydrogen or halo, the method of embodiment 64.

[0386] 89.R 8 The method of embodiment 64, wherein R is independently hydrogen, halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino.

[0387] 90.R 8 The method of embodiment 64, wherein R is hydrogen.

[0388] As will be apparent to those skilled in the art, many modifications and variations of the present invention can be made without departing from its spirit and scope. The specific embodiments described herein are provided by way of example only, and the present invention is limited only by the requirements of the appended claims and the full scope of equivalents of the rights given by such claims. Such modifications are intended to be included within the scope of the appended claims.

[0389] All references, patents, and non-patents cited herein are hereby incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference for all purposes.

Claims

1. Formula (A) 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof, 【Chemistry 2】 represents a single bond or a double bond; m or n is independently an integer of 0, 1, 2, or 3; p is 0 or 1; Z １ Or Z ２ are independently O, S, and SO ２ , N.R. ３ , or C.R. ５ R ６ and Y １ -Y １０ each independently represents N or CR ７ and Y １ -Y １０ is N, provided that 【Chemistry 3】 The parts come together 【Chemistry 4】 forming a portion Y １ -Y ５ One of the is N and the other is Y ８ -Y １０ One of the is N and the remaining Y １ -Y １０ When each of Y is CH, １ -Y ５ together with the amidine substituent, Y ８ -Y １０ forming an amidine-substituted pyridine ring different from the amidine-substituted pyridine ring formed by R １ and R ２ are each independently hydrogen or halo; Or R １ R ２ together with R form a saturated, unsaturated or partially unsaturated 3- to 9-membered cyclic group, which is optionally substituted with halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino, with the proviso that R １ R ２ together with Z to form a phenyl group; １ and Z ２ is O and Y １ -Y ５ One of the is N and the other is Y ８ -Y １０ One of the is N and the remaining Y １ -Y １０ When each of Y is CH, １ -Y ５ together with the amidine substituent, Y ８ -Y １０ forming an amidine-substituted pyridine ring different from the amidine-substituted pyridine ring formed by R ３ is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl; R ４ is hydrogen, halo, cycloalkyl, aryl, or heteroaryl; R ５ Or R ６ are independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, or amino; Or, R ５ R ６ together with R ７ are independently hydrogen, halo, or amidine (-Am) 【Chemistry 5】 (It is.)

2. The compound is represented by formula (I) 【Chemistry 6】 or a pharma- ceutically acceptable salt thereof.

3. The compound has formula (II): 【Chemistry 7】 or a pharma- ceutically acceptable salt thereof, X is independently N or CR ７ with the proviso that at least one X is N; R ７ are independently hydrogen or halo.

4. The compound has formula (III): 【Chemistry 8】 or a pharma- ceutically acceptable salt thereof, X is independently N or CR ７ with the proviso that at least one X is N; R ７ are independently hydrogen or halo.

5. The compound has formula (IV): 【Chemistry 9】 or a pharma- ceutically acceptable salt thereof, X is independently N or CR ７ with the proviso that at least one X is N; R ７ are independently hydrogen or halo.

6. The compound has formula (V): 【Chemistry 10】 or a pharma- ceutically acceptable salt thereof, ８ is hydrogen, halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino.

7. The compound has formula (VI): 【Chemistry 11】 or a pharma- ceutically acceptable salt thereof, X is independently N or CR ７ with the proviso that at least one X is N; R ８ is hydrogen, halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino.

8. The compound has formula (VII): 【Chemistry 12】 or a pharma- ceutically acceptable salt thereof, X is independently N or CR ７ with the proviso that at least one X is N; R ８ is hydrogen, halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino.

9. The compound has formula (VIII): 【Chemistry 13】 or a pharma- ceutically acceptable salt thereof, X is independently N or CR ７ with the proviso that at least one X is N; R ８ is hydrogen, halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino.

10. The compound according to any one of claims 1 to 5, wherein m is 1 and n is 1, or a pharma- ceutically acceptable salt thereof.

11. The compound according to any one of claims 1 to 5, wherein m is 1 and n is 0, or a pharma- ceutically acceptable salt thereof.

12. The compound according to any one of claims 1 to 5, wherein m is 0 and n is 1, or a pharma- ceutically acceptable salt thereof.

13. 6. The compound according to any one of claims 1 to 5, wherein m is 1 and n is 2, or a pharma- ceutically acceptable salt thereof.

14. The compound according to any one of claims 1 to 5, wherein m is 2 and n is 1, or a pharma- ceutically acceptable salt thereof.

15. 6. The compound according to any one of claims 1 to 5, wherein m is 2 and n is 2, or a pharma- ceutically acceptable salt thereof.

16. The compound according to any one of claims 1 to 5, wherein m is 0 and n is 0, or a pharma- ceutically acceptable salt thereof.

17. R ８ 10. The compound of any one of claims 1 and 6-9, wherein is independently hydrogen, halo, cyano, alkyl, cycloalkyl, aryl, heteroaryl, or amino, or a pharma- ceutically acceptable salt thereof.

18. R ８ 18. The compound of claim 17, or a pharma- ceutically acceptable salt thereof, wherein is hydrogen.

19. Z １ Or Z ２ is independently selected from the group consisting of N, O, and S, or a pharma- ceutically acceptable salt thereof.

20. Z １ Or Z ２ is independently S; or a pharma- ceutically acceptable salt thereof.

21. Z １ Or Z ２ is independently O; or a pharma- ceutically acceptable salt thereof.

22. Z １ Or Z ２ is independently N; or a pharma- ceutically acceptable salt thereof.

23. Z １ Or Z ２ became independent and became NR ３ and R ３ 20. The compound of claim 19, or a pharma- ceutically acceptable salt thereof, wherein is hydrogen.

24. Z １ Or Z ２ became independent and became NR ３ and R ３ 20. The compound of claim 19, or a pharma- ceutically acceptable salt thereof, wherein is selected from the group consisting of alkyl, cycloalkyl, aryl, and heteroaryl.

25. Z １ Or Z ２ is independent, NR ３ Or CR ５ R ６ 19. The compound of any one of claims 1 to 18, or a pharma- ceutically acceptable salt thereof.

26. Z １ is NR ３ and R ３ is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl; Z ２ is CR ５ R ６ and R ５ Or R ６ are independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or amino, or R ５ R ６ and R 1 and R 2 are together formed a saturated or partially unsaturated 3- to 9-membered ring, or a pharma- ceutically acceptable salt thereof.

27. Z １ Or Z ２ became independent and ５ R ６ and R ５ Or R ６ are independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, or amino, or R ５ R ６ and R 1 and R 2 are together formed a saturated or partially unsaturated 3- to 9-membered ring, or a pharma- ceutically acceptable salt thereof.

28. R ５ Or R ６ 26. The compound of claim 25, or a pharma- ceutically acceptable salt thereof, wherein: is independently hydrogen.

29. Z １ Or Z ２ became independent and became NR ３ and R ３ The compound according to any one of claims 1 to 18, or a pharma- ceutically acceptable salt thereof, wherein is hydrogen, alkyl, cycloalkyl, aryl, or heteroaryl.

30. Z １ Or Z ２ are independently O, S, or SO ２ 19. The compound according to any one of claims 1 to 18, wherein:

31. Y ３ and Y ８ or a pharma- ceutically acceptable salt thereof.

31. The compound of claim 1, wherein:

32. Y ３ and Y ７ or a pharma- ceutically acceptable salt thereof.

31. The compound of claim 1, wherein:

33. Y ２ and Y ７ or a pharma- ceutically acceptable salt thereof.

31. The compound of claim 1, wherein:

34. R １ and R ２ or a pharma- ceutically acceptable salt thereof. The compound according to any one of claims 1 to 33, wherein:

35. R １ R ２ together with a saturated, unsaturated or partially unsaturated 3-9 membered cyclic group, for example 【Chemistry 14】 34. The compound according to any one of claims 1 to 33, or a pharma- ceutically acceptable salt thereof, which forms:

36. R １ R ２ or a pharma- ceutically acceptable salt thereof, which is combined with R to form a 5-, 6-, or 7-membered cycloalkyl.

37. R １ R ２ or a pharma- ceutically acceptable salt thereof, which is combined with

38. R １ R ２ or a pharma- ceutically acceptable salt thereof, which is combined with

39. Y １、２、４、５、６、及び８ is CR ７ (e.g., —CH), and Y ２ is N and Y ３ and Y ７ or a pharma- ceutically acceptable salt thereof, wherein:

40. Y １、４、５、６、及び７ is -CH, and Y ２ is N and Y ３ and Y ８ is CR ７ and R ７ The compound according to any one of claims 1 to 38, or a pharma- ceutically acceptable salt thereof, wherein is an amidine.

41. Y １、４、５、６、及び８ is -CH, and Y ３ is N and Y ２ and Y ７ is CR ７ and R ７ The compound according to any one of claims 1 to 38, or a pharma- ceutically acceptable salt thereof, wherein is an amidine.

42. Y １、４、５、６、及び８ is -CH, and Y ３ is N and Y ２ and Y ７ is CR ７ and R ７ is amidine, m is 1, and n is 0; or a pharma- ceutically acceptable salt thereof.

43. Y １、４、５、及び６ is -CH, and Y ３ and Y ８ is N and Y ２ and Y ７ is CR ７ and R ７ is amidine, m is 1, and n is 0; or a pharma- ceutically acceptable salt thereof.

44. The compound is 5-(5-(4-carbamimidoylphenoxy)pentyloxy)picolinimidamide; 6-((5-(4-carbamimidoylphenoxy)pentyl)oxy)nicotinimidamide; 5-((5-(4-carbamimidoylphenoxy)pentyl)oxy)pyrimidine-2-carboximidamide; 5-((5-(4-carbamimidoylphenoxy)pentyl)oxy)pyrazine-2-carboximidamide; 5-(4-(4-carbamimidoylphenoxy)butoxy)picolinimidamide; 5-(4-(4-carbamimidoylphenoxy)butoxy)picolinimidamide 5,5'-(pentane-1,5-diylbis(oxy))bis(pyrazine-2-carboximidamide); 6,6'-(heptane-1,7-diyl)dipicolinimidamide; 5,5'-(heptane-1,7-diyl)dinicotinimidamide; 6,6'-(heptane-1,7-diyl)dinicotinimidamide; 5-(5-(3-carbamimidoylphenoxy)pentyloxy)picolinimidamide; 4-({5-[(6-cyanopyridin-3-yl)oxy]pentyl}oxy)pyridine-2-carbonitrile; 5-(((1r,4r)-4-(4-carbamimidoylphenoxy)cyclohexyl)oxy)picolinimidamide; 5-(((1s,4s)-4-(4-carbamimidoylphenoxy)cyclohexyl)oxy)picolinimidamide; 4-(5-(3-carbamimidoylphenoxy)pentyloxy)picolinimidamide; 5,5'-(butane-1,4-diylbis(oxy))dipicolinimidamide; 5-(3-(4-carbamimidoylphenoxy)propoxy)picolinimidamide; 5-{2-[(1R,3S)-3-[2-(4-carbamimidoylphenyl)ethyl]cyclohexyl]ethyl}pyridine-2-carboximidamide; 4-{[5-(4-carbamimidoylphenoxy)pentyl]oxy}pyridine-2-carboximidamide; 5-({5-[(6-carbamimidoylpyridin-3-yl)oxy]pentyl}oxy)pyridine-2-carboximidamide; and 2. The compound of claim 1, or a pharma- ceutically acceptable salt thereof, selected from the group consisting of 4-({5-[(2-carbamimidoylpyridin-4-yl)oxy]pentyl}oxy)pyridine-2-carboximidamide.

45. A compound having the following structure: 【Chemistry 15】 or a pharma- ceutically acceptable salt thereof.

46. A compound having the following structure: 【Chemistry 16】 or a pharma- ceutically acceptable salt thereof.

47. 47. A pharmaceutical composition comprising a compound according to any one of claims 1 to 46, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

48. A method for treating cancer, comprising administering to a subject suffering from cancer an effective amount of a compound according to any one of claims 1 to 46 or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to claim 47.

49. 49. The method of claim 48, wherein the cancer is selected from the group consisting of liver cancer, bile duct cancer, osteosarcoma, melanoma, breast cancer, renal cancer, prostate cancer, gastric cancer, colorectal cancer, thyroid cancer, head and neck cancer, ovarian cancer, pancreatic cancer, neurological cancer, lung cancer, uterine cancer, leukemia, and lymphoma.

50. 50. The method of claim 49, wherein the cancer is liver cancer.

51. 50. The method of claim 49, wherein the cancer is cholangiocarcinoma.

52. 50. The method of claim 49, wherein the cancer is prostate cancer.

53. 50. The method of claim 49, wherein the cancer is pancreatic cancer.

54. 50. The method of claim 49, wherein the cancer is lung cancer.

55. 50. The method of claim 49, wherein the cancer is small cell lung cancer.

56. 50. The method of claim 49, wherein the cancer is non-small cell lung cancer.

57. 50. The method of claim 49, wherein the cancer is breast cancer.

58. 50. The method of claim 49, wherein the cancer is colorectal cancer.

59. 50. The method of claim 49, wherein the cancer is renal cancer.

60. 50. The method of claim 49, wherein the cancer is a solid tumor.

61. The compound of formula (A) or a pharma- ceutically acceptable salt thereof may be administered orally, intravenously, or subcutaneously to the subject (e.g., a human patient) at a dose of about 0.5 mg / kg, 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, about 100 mg / kg 50. The method of claim 49, wherein the medicament is administered at a dose of about 110 mg / kg, about 120 mg / kg, about 130 mg / kg, about 140 mg / kg, about 150 mg / kg, about 160 mg / kg, about 170 mg / kg, about 180 mg / kg, about 190 mg / kg, about 200 mg / kg, about 210 mg / kg, about 220 mg / kg, about 230 mg / kg, about 240 mg / kg, about 250 mg / kg, about 260 mg / kg, about 270 mg / kg, about 280 mg / kg, about 290 mg / kg, about 300 mg / kg, about 350 mg / kg, about 400 mg / kg, about 450 mg / kg, about 500 mg / kg, or about 600 mg / kg.

62. 50. The method of claim 49, wherein the subject is a human patient.

63. 50. The method of claim 49, wherein the compound of formula (A) or a pharma- ceutically acceptable salt thereof is administered orally to the human patient.

64. 50. The method of claim 49, wherein the subject is administered about 1 mg / kg to about 200 mg / kg daily.

65. 50. The method of claim 49, wherein the subject is administered about 1 mg / kg to about 100 mg / kg daily.

66. 49. The method of claim 48, wherein the subject is administered about 1 mg / kg to about 50 mg / kg daily.

67. 50. The method of claim 49, wherein the subject is administered about 0.5 mg / kg to about 50 mg / kg daily.

68. 50. The method of claim 49, wherein the subject is administered about 2 mg / kg daily.

Images