(12) United States Patent (10) Patent No.: US 9.415,047 B2 Zhou Et Al

US0094.15047B2

(12) United States Patent (10) Patent No.: US 9.415,047 B2 Zhou et al. (45) Date of Patent: Aug. 16, 2016

(54) USE OF BENZO FIVE-MEMBERED 31/4545; A61K 31/506; C07D 209/08; C07D NITROGEN HETEROCYCLIC PPERAZINE 231/56; C07D 235/08; C07D 235/14: CO7D OR PIPERDINEDERVATIVES 235/22; C07D401/06; C07D 401/12 (71) Applicants: Liaoning Emmy Biological USPC ...... 514/252.13, 254 Pharmaceutical Co., Ltd., Liaoning See application file for complete search history. (CN); Shenyang Emmy Pharmaceutical Research Institute (56) References Cited Co., Ltd., Liaoning (CN) U.S. PATENT DOCUMENTS (72) Inventors: Yan Zhou, Liaoning (CN): Lirong Zhang, Liaoning (CN); Jie Zhou, 3,362.956 A * 1/1968 Archer ...... CO7D 209/60 Liaoning (CN), Xin Zhou, Liaoning 544,225 3,472,854. A * 10/1969 Archer ...... CO7D 209/60 (CN) 514,906 (73) Assignees: Liaoning Emmy Biological (Continued) Pharmaceutical Co., Ltd., Liaoning (CN); Shenyang Emmy FOREIGN PATENT DOCUMENTS Pharmaceutical Research Institute Co., Ltd., Liaoning (CN) CN 1154693. A 7/1997 CN 1944404 A 4/2007 (*) Notice: Subject to any disclaimer, the term of this CN 101759693. A 6, 2010 patent is extended or adjusted under 35 U.S.C. 154(b) by 0 days. OTHER PUBLICATIONS (21) Appl. No.: 14/647,408 Alessandro Boido et al., “Synthesis and pharmacological evaluation (22) PCT Fled: Nov. 25, 2013 of aryl/heteroaryl piperazinyl alkyl benzotriazoles as ligands for some serotonin and dopamine receptor subtypes.” IL FARMACO, (86) PCT NO.: PCT/CN2O13/OO1442 2001, vol. 56, pp. 263-275.* S371 (c)(1), (Continued) (2) Date: May 26, 2015 (87) PCT Pub. No.: WO2O14/07915S Primary Examiner — Kamal Saeed PCT Pub. Date: May 30, 2014 Assistant Examiner — Janet L. Coppins (74) Attorney, Agent, or Firm — Hultquist, PLLC; Steven J. (65) Prior Publication Data Hultquist US 2015/O297586 A1 Oct. 22, 2015 (30) Foreign Application Priority Data (57) ABSTRACT Nov. 26, 2012 (CN) ...... 2012 1 0486619 This invention relates to the use of a compound informula (I) Nov. 26, 2012 (CN) ...... 2012 1 0486967 and its salts acceptable pharmaceutically in preparation of Nov. 26, 2012 (CN) ...... 2012 1 04871 28 vasodilative drugs: (51) Int. Cl. (I) A6 IK3I/496 (2006.01) A6 IK3 L/454 (2006.01) / x (Continued) (52) U.S. Cl. CPC ...... A6 IK3I/496 (2013.01); A61 K3I/454 R-X N-Y-N-A (2013.01); A61 K3I/4545 (2013.01); (Continued) Wherein, R. R. X, Y, A and B are defined in the invention. (58) Field of Classification Search CPC ...... A61K 31/496; A61K 31/454: A61 K 19 Claims, 11 Drawing Sheets

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(51) Int. Cl. (56) References Cited A6 IK3I/498 (2006.01) C07D 209/08 (2006.01) U.S. PATENT DOCUMENTS CO7D 23L/56 (2006.01) 2010/0329978 A1 12/2010 McCurdy et al. CO7D 403/2 (2006.01) 2011/0306638 A1 12/2011 Li et al. CO7D 40/4 (2006.01) CO7D 405/4 (2006.01) OTHER PUBLICATIONS CO7D 413/4 (2006.01) PubMed abstract of Dabire et al., “Central 5-hydroxytyptamine A6 IK3 L/4545 (2006.01) (5-HT) receptors in blood pressure regulation.” Therapie, 1991, vol. CO7D 235/08 (2006.01) 46(6), pp. 421-429.* CO7D 40/06 (2006.01) Boido, A., et al., “Synthesis and pharmacological evaluation of aryl/ heteroaryl piperazinyl alkyl benzotriazoles as ligands for some CO7D 40/12 (2006.01) serotonin and dopamine receptor Subtypes”. “IL Farmaco'. Apr. CO7D 405/2 (2006.01) 2001, pp. 263-275, vol. 56. CO7D 413/2 (2006.01) Boido, A., et al., “Alpha1- and alpha2-adrenoreceptor antagonist C07D 417/12 (2006.01) profiles of 1- and 2-omega-(4-arylpiperazin-1-yl)alkyl-1,2,3- CO7D 417/4 (2006.01) benzotriazoles”, “Chemistry & Biodiversity, Oct. 2005, pp. 1290 1304, vol. 2. CO7D 249/8 (2006.01) Caliendo, G., et al., “Synthesis and biological activity of A6 IK3I/506 (2006.01) benzotriazole derivatives structurally related to trazodone”, “Euro C07D 235/10 (2006.01) pean Journal of Medicinal Chemistry”. Jan. 1, 1995, pp. 77-84, vol. CO7D 235/4 (2006.01) 30. C07D 235/22 (2006.01) Caliendo, G., et al., “Structureaffinity relationship studies on A6 IK 45/06 (2006.01) benzotriazole derivatives binding to 5-HT receptor subtypes”, “Euro 52) U.S. C pean Journal of Medicinal Chemistry”, 1996, pp. 207-213, vol. 31. (52) AV e. we Hoyer, D., et al., “International Union of Pharmacology classification CPC ...... A61 K3I/498 (2013.01); A61 K3I/506 of receptors for 5-hydroxytryptamine (Serotonin)”, “Pharmacologi (2013.01); A61K 45/06 (2013.01); C07D cal Reviews”, Jun. 1994, pp. 157-203, vol. 46, No. 2. 209/08 (2013.01); C07D 231/56 (2013.01); Mesangeau, C., et al., “Synthesis and pharmacological evaluation of C07D 235/08 (2013.01); C07D 235/10 indole-based sigma receptor ligands”, “European Journal of Medici nal Chemistry”, Aug. 29, 2011, pp. 5154-5161, vol. 46. (2013.01); C07D 235/14 (2013.01); C07D Mokrosz, M., et al., “Structure-activity relationship studies of CNS 235/22 (2013.01); C07D 249/18 (2013.01): agents, Part32: Effect of structural modifications in 1-arylpiperazine C07D401/06 (2013.01); C07D401/12 derivatives on alpha(1) -adrenoreceptor affinity”, “Arch. Pharm. (2013.01); C07D401/14 (2013.01); C07D (Weinheim).”. Jun. 1997, pp. 177-180, vol. 330. 403/12 (2013.01); C07D405/12 (2013.01); Co-pending Unpublished U.S. Appl. No. 147647.378, filed May 25, C07D405/14 (2013.01); C07D 413/12 2015. (2013.01); C07D 413/14 (2013.01); C07D 417/12 (2013.01); C07D 417/14 (2013.01) * cited by examiner U.S. Patent Aug. 16, 2016 Sheet 1 of 11 US 9.415,047 B2

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-log C (5-HT, moll-1 Figare 24 US 9,415,047 B2 1. 2 USE OF BENZO FIVE-MEMBERED (C-C alkyl), alkyl parts in the above said groups are substi NITROGEN HETEROCYCLIC PPERAZINE tuted by anyone or more halogen atoms. OR PIPERDINEDERVATIVES A, B and X respectively indicate CH or N. R indicates H. halogen, CN, C-C alkyl, C-C alkoxyl, CROSS-REFERENCE TO RELATED 5 CHO, CO(C-C alkyl). COO(C-C alkyl), COOH, NO, APPLICATIONS NH, NH(C-C alkyl), N(C-C alkyl), SH, S(C-C alkyl). —S(O) (C-C alkyl). —S(O)H or —S(O)(C-C alkyl). This application is a U.S. national phase under the provi alkyl parts in the above said groups are substituted by anyone sions of 35 U.S.C. S371 of International Patent Application or more halogenatoms. If R indicates poly Substituted group, No. PCT/CN13/01442 filed Nov. 25, 2013, which in turn 10 claims priority of Chinese Patent Application No. R is independently chosen from halogen, CN, C-C alkyl, 201210487128.6 filed Nov. 26, 2012, Chinese Patent Appli C-C alkoxyl, CHO, CO(C-C alkyl). COO(C-C alkyl). cation No. 201210486967.6 filed Nov. 26, 2012, and Chinese COOH, NO, NH, NH(C-C alkyl), N(C-C alkyl), SH, Patent Application No. 2012.10486619.9 filed Nov. 26, 2012. S(C-C alkyl). —S(O) (C-C alkyl). —S(O)H or —S(O) The disclosures of Such international patent application and 15 (C-C alkyl), alkyl parts in the above said groups are substi Chinese priority patent applications are hereby incorporated tuted by anyone or more halogen atoms. herein by reference in their respective entireties, for all pur Y indicates Saturated or unsaturated Straight or branched poses. hydrocarbon chain composed of 1-8 carbonatoms substituted by anyone or more halogen atoms, in which anyone or more FIELD OF THE INVENTION 2O carbonatoms are Substituted by hetero atoms Such as oxygen, Sulfur or nitrogen. The present invention relates to use of benzo five-mem bered nitrogen heterocyclic piperazine or piperidine deriva ILLUSTRATION BY ACCOMPANYING tives in preparation of vasodilative drugs. FIGURES 25 BACKGROUND OF THE INVENTION FIG. 1 illustrates the accumulated concentration effect curve of vasodilative effects of compound II-2 (10-10' Currently there are various categories of vasodilative drugs mol-L") versus vasoconstrictive effects of adrenaline (10 available in clinical field, e.g., C. receptor blockers, including mol-L") on excised blood vessels from rabbits. furazosin, doxazosin and teraZosin, etc., which have obvious 30 FIG. 2 illustrates the accumulated concentration effect first dose effects or orthostatic hypotension, so their extensive curve of vasodilative effects of compound II-2 (10-10' application is limited inclinical practice. Ca" channel block mol-L") versus vasoconstrictive effects of high potassium ers, including amlodipine, nifedipine and felodipine, etc. cur concentration (60 mmol-L") on excised blood vessels from rently, which are still extensively applied in clinical practice, rabbits. but also with risks of heart Suppression. 35 FIG. 3 illustrates the accumulated concentration effect Therefore, it is still necessary to develop new vasodilative curve of resistance of compound II-2 (3x107 mol/L) to vaso drugs, improve efficacy, reduce drug resistance or minimize constrictive effects of noradrenaline NA (10-10" mol/L) drug toxicity, to satisfy clinical demands of different patients. on excised blood vessels from rabbits. FIG. 4 illustrates the accumulated concentration effect CONTENTS OF THE INVENTION 40 curve of resistance of positive reference drug doxazosin (107 mol/L) to vasoconstrictive effects of noradrenaline NA This invention offers the use of a compound in formula (I) (10-6x10 mol/L) on excised blood vessels from rabbits. and its salts acceptable pharmaceutically in preparation of FIG. 5 illustrates the accumulated concentration effect vasodilative drugs: curve of resistance of compound II-2 (10 mol/L) to vaso 45 constrictive effects of CaCl (10-10° mol/L) on excised blood vessels from rabbits. (I) FIG. 6 illustrates the accumulated concentration effect curve of resistance of amlodipine (107 mol/L) to vasocon strictive effects of CaCl, (10-10° mol/L) on excised blood 50 vessels from rabbits. FIG. 7 illustrates the accumulated concentration effect R-X N-Y-N( A w n2 curve of resistance of compound II-2 (3x10 mol/L) to vaso constrictive effects of 5-hydroxytryptamine (107-3x10' mol/L) on excised blood vessels from rabbits. Wherein, 55 FIG. 8 illustrates the accumulated concentration effect R indicates aromatic groups oralicyclic groups with mono curve of vasodilative effects of compound II-3 (10-3x10 or polysubstituted by R, wherein, mol-L") versus vasoconstrictive effects of adrenaline (10 R indicates H. halogen, CN, C-C alkyl, C-C alkoxyl, mol-L") on excised blood vessels from rabbits. CHO, CO(C-C alkyl), COO(C-C alkyl), COOH, NO, FIG. 9 illustrates the accumulated concentration effect NH, NH(C-C alkyl), N(C-C alkyl), SH, S(C-C alkyl), 60 curve of vasodilative effects of compound II-3 (10-3x10 —S(O) (C-C alkyl). —S(O)H or —S(O), (C-C alkyl). mol-L") versus vasoconstrictive effects of high potassium alkyl parts in the above said groups are Substituted by anyone concentration (60 mmol-L") on excised blood vessels from or more halogenatoms. If R indicates polysubstituted group, rabbits. R is independently chosen from halogen, CN, C-C alkyl, FIG. 10 illustrates the accumulated concentration effect C-C alkoxyl, CHO, CO(C-C alkyl), COO(C-C alkyl), 65 curve of vasodilative effects of compound II-31 (10-10 COOH, NO, NH, NH(C-C alkyl), N(C-C alkyl), SH, mol-L") versus vasoconstrictive effects of adrenaline AD S(C-C alkyl). —S(O) (C-C alkyl). —S(O)H or —S(O) (10 mol-L") on excised blood vessels from rabbits. US 9,415,047 B2 3 FIG. 11 illustrates the accumulated concentration effect curve of vasodilative effects of compound II-31 (3x107-3x (I) 10 mol-L) versus vasoconstrictive effects of high potas sium concentration (60 mmol-L") on excised blood vessels from rabbits. FIG. 12 illustrates the accumulated concentration effect curve of resistance of compound II-31 (3x10" mol/L) and positive reference drug doxazosin (107 mol/L) to vasocon strictive effects of noradrenaline NA (3x107-10 mol/L) on excised blood vessels from rabbits. 10 Wherein, FIG. 13 illustrates the accumulated concentration effect R indicates aromatic groups oralicyclic groups with mono curve of resistance of compound II-31 (10 mol/L) and or polysubstituted by R, wherein, amlodipine (107 mol/L) to vasoconstrictive effects of CaCl, R indicates H. halogen, CN, C-C alkyl, C-C alkoxyl, (10-3x10" mol/L) on excised blood vessels from rabbits. 15 CHO, CO(C-C alkyl). COO(C-C alkyl), COOH, NO, FIG. 14 illustrates the accumulated concentration effect NH, NH(C-C alkyl), N(C-C alkyl), SH, S(C-C alkyl). curve of resistance of compound II-31 (3x10" mol/L) to —S(O) (C-C alkyl). —S(O)H or —S(O)(C-C alkyl). vasoconstrictive effects of 5-hydroxytryptamine (10-3x alkyl parts in the above said groups are substituted by anyone 10 mol/L) on excised blood vessels from rabbits. or more halogenatoms. If R indicates poly Substituted group, FIG. 15 illustrates the accumulated concentration effect R is independently chosen from halogen, CN, C-C alkyl, curve of vasodilative effects of compound II-29 (10-3x10 C-C alkoxyl, CHO, CO(C-C alkyl). COO(C-C alkyl). mol-L") versus vasoconstrictive effects of adrenaline AD COOH, NO, NH, NH(C-C alkyl), N(C-C alkyl), SH, (10 mol-L") on excised blood vessels from rabbits. S(C-C alkyl). —S(O) (C-C alkyl). —S(O)H or —S(O) FIG. 16 illustrates the accumulated concentration effect (C-C alkyl), alkyl parts in the above said groups are substi curve of vasodilative effects of compound II-29 (107-3x10 25 tuted by anyone or more halogen atoms. mol-L") versus vasoconstrictive effects of high potassium A, B and X respectively indicate CH or N. concentration (60 mmol-L") on excised blood vessels from R indicates H. halogen, CN, C-C alkyl, C-C alkoxyl, rabbits. CHO, CO(C-C alkyl), COO(C-C alkyl), COOH, NO, FIG. 17 illustrates the accumulated concentration effect NH, NH(C-C alkyl), N(C-C alkyl), SH, S(C-C alkyl). curve of vasodilative effects of compound II-85 (10'-10° 30 —S(O) (C-C alkyl) or —S(O)2(C-C alkyl), alkyl parts in mol-L") versus vasoconstrictive effects of adrenaline (10 the above said groups are substituted by anyone or more mol-L") on excised blood vessels from rabbits. halogen atoms. If R indicates polysubstituted group, R is FIG. 18 illustrates the accumulated concentration effect independently chosen from halogen, CN, C-C alkyl, C-C, curve of vasodilative effects of compound II-85 (10'-10° alkoxyl, CHO, CO(C-C alkyl), COO(C-C alkyl), COOH, mol-L") versus vasoconstrictive effects of noradrenaline 35 (10 mol-L") on excised blood vessels from rabbits. NO, NH, NH(C-C alkyl), N(C-C alkyl), SH, S(C-C, FIG. 19 illustrates the accumulated concentration effect alkyl). —S(O) (C-C alkyl). —S(O)H or —S(O)(C-C, curve of vasodilative effects of compound II-85 (109-10 alkyl), alkyl parts in the above said groups are substituted by mol-L") versus vasoconstrictive effects of high potassium anyone or more halogen atoms. concentration (60 mmol-L") on excised blood vessels from 40 Y indicates Saturated or unsaturated Straight or branched rabbits. hydrocarbon chain composed of 1-8 carbonatoms substituted FIG. 20 illustrates the accumulated concentration effect by anyone or more halogen atoms, in which anyone or more curve of resistance of compound II-85 (10 mol/L) to vaso carbonatoms are Substituted by hetero atoms Such as oxygen, constrictive effects of phenephrine (10-6x10 mol/L) on Sulfur or nitrogen. excised blood vessels from rabbits. 45 Preferably, R informula (I) in this invention is the mono or FIG. 21 illustrates the accumulated concentration effect polysubstituted group in the said benzo five-membered nitro curve of resistance of doxazosine mesylate (10 mol/L) to gen heterocyclic ring, e.g., R indicates mono-, di- or trisub vasoconstrictive effects of phenephrine (10-3x10 mol/L) stituted group, etc. Rindicates the group linked to any carbon on blood vessels from rabbits. atoms on benzo five-membered nitrogen heterocyclic ring, FIG. 22 illustrates the accumulated concentration effect 50 e.g., when A (or B) is CH atom, to which R can also be linked curve of resistance of compound II-85 (10 mol/L) to vaso It should be understood that, in term “aromatic group' used constrictive effects of CaCl (10-10° mol/L) on excised in this document, at least one ring should be a Cs2 hydro blood vessels from rabbits. carbon mono orbicyclic ring of aromatic rings, in which one FIG. 23 illustrates the accumulated concentration effect or more carbon atoms are substituted by hetero atoms of curve of resistance of amlodipine (107 mol/L) to vasocon 55 oxygen, Sulfur and nitrogen. The examples of aromatic strictive effects of CaCl (10-10° mol/L) on excised blood groups include aryl and hetero aryl groups, such as phenyl, vessels from rabbits. naphthyl, benzothiazolyl, benzisothiazolyl, benzoxazolyl, FIG. 24 illustrates the accumulated concentration effect benzisoxazolyl, benzimidazoyl, benzopyrazol, benzofuranyl. curve of resistance of compound II-85 (107 mol/L) to vaso benzopyrimidinyl, benzopyridyl, quinoxalinyl, furyl, pyridyl constrictive effects of 5-hydroxytryptamine (10-10 mol/ 60 or pyrimidinyl groups. L) on excised blood vessels from rabbits. It should be understood that, in term “alicyclic group' used in this document, C-2 Saturated hydrocarbon monocycle or SPECIFIC OPERATING PROCEDURE bicycle, in which one or more carbonatoms are substituted by hetero atoms of oxygen, Sulfur and nitrogen. The examples of This invention offers the use of a compound in formula (I) 65 alicyclic group include cyclopropyl, cyclobutyl, cyclopentyl, and its salts acceptable pharmaceutically in preparation of cyclohexyl, cycloheptyl, tetrahydrofuranyl, piperidyl or pip vasodilative drugs: eraZidinyl, etc. US 9,415,047 B2 5 6 Except for otherwise specified, the term “halogen used in COOCH in which alkyl parts are substituted 1-3 halogen this document indicates fluorine, chlorine, bromine or iodine. atoms. Further preferably, R indicates H. F. Cl, COCH, Term “alkyl used in this document includes straight or C-C alkyl or C-C alkoxyl in which alkyl parts are substi branch alkyl. Examples of the said "C-C alkyl group tuted 1-3 halogen atoms. Further more preferably, R indi include methyl, ethyl, n-propyl, isopropyl. n-butyl, isobutyl, cates H. F. C1, CN, CF, CH, OCH or COCH. Again pref teriary butyl, n-pentyl, isopentyl, teriary pentyl, n-hexyl, iso erably, R indicates H. F. C1, CN, CF, CH or OCH. Most hexyl, etc. preferably, when X is N, R indicates H. F. C1 or OCH. Term “alkoxyl used in this document indicates —O-alkyl, Particularly preferably, when X is CH, R indicates H. F or wherein alkyl groups include Straight or branchalkyl groups. CF, R is more preferably F. If R indicates polysubstituted Examples of the said "C-C alkoxyl groups include meth 10 group, R is independently chosen from the above said oxyl, ethoxyl, propoxyl, butoxyl, pentyloxyl and hexaoxyl, groups. etc. A, B and X respectively indicate CH or N. Preferably, A As used herein, definitions including general, preferable, and Ball indicate N. more preferable, further preferable, particularly preferable 15 R indicates H. halogen, CN, C-C alkyl, C-C alkoxyl, and most preferable can be mutually combined. CHO, CO(C-C alkyl), COO(C-C alkyl), COOH, NO, In an embodiment, this invention offers the use of a com NH, NH(C-C alkyl), N(C-C alkyl), SH, S(C-C alkyl). pound informula (I) and its salts acceptable pharmaceutically —S(O) (C-C alkyl). —S(O)H or - S(O), (C-C alkyl). in preparation of vasodilative drugs: alkyl parts in the above said groups are substituted by anyone or more halogen atoms. Preferably, R indicates H. halogen, CN, C-C alkyl, C-C alkoxyl, CHO, CO(C-C alkyl). (I) COO(C-C alkyl), COOH or NO, alkyl parts in the above said groups are Substituted by anyone or more (e.g., 1-3 atoms) halogenatoms. More preferably, R indicates H, F, Cl, 25 Br, CN, NO, C-C alkyl or C-C alkoxyl, CHO, COCH or COOCH in which alkyl parts are substituted 1-3 halogen atoms. Further preferably, R indicates H. F. C1, CN, CHO, COCH, COOCH or C-C alkyl or C-C alkoxyl in which alkyl parts are substituted 1-3 halogen atoms. Further more Wherein, 30 R indicates aromatic groups oralicyclic groups with mono preferably, R indicates H, F, C1, CN, CF, CH, OCH, CHO, or polysubstituted by R, wherein, COCH or COOCH. Particularly preferably, when X indi The said aromatic groups preferably indicates phenyl, cates N, R indicates H. FC1, CN, CH or COOCH R more naphthyl, benzo five or six membered heterocyclic rings with preferably indicates H. F. C1 or CH. Particularly preferably, heteroatoms of N, S or O, or five or six membered unsaturated 35 when X indicates CH, R indicates H or OCH. Most prefer heterocyclic rings. More preferably phenyl, naphthyl, ben ably, R indicates H. If R indicates polysubstituted group, R Zothiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, is independently chosen from the above said groups. benzimidazoyl, benzopyrazol, benzofuranyl, benzopyrimidi Y indicates Saturated or unsaturated Straight or branched nyl, benzopyridyl, quinoxalinyl, furyl, pyridyl or pyrimidinyl hydrocarbon chain composed of 1-8 carbonatoms substituted groups. Further preferably phenyl, benzisoxazolyl, ben 40 by 1-3 halogenatoms, in which anyone or more carbonatoms Zisothiazolyl, benzopyrazol, benzofuranyl, naphthyl, furyl, are substituted by hetero atoms Such as oxygen, Sulfur or pyridyl or pyrimidinyl, quinoxalinyl groups. Particularly nitrogen. Preferably, Yindicates unsubstituted saturated alkyl preferably phenyl, benzisoxazolyl, benzisothiazolyl, benzo group composed of 1-8 carbon atoms, or unsubstituted Satu furanyl, quinoxalinyl, pyrimidinyl groups. Particularly pref rated alkyl group composed of 1-8 carbonatoms in which one erably phenyl, benzisoxazolyl groups. Most preferably, when 45 carbon atom is replaced by oxygen or sulfur, e.g., —C, the said aromatic group is phenyl, X indicates N. Most pref alkylidene-O . More preferably, Yindicates methylene, eth erably, when the said aromatic group is benzisoxazolyl. X ylidene, propylidene, butylidene, pentylidene, hexylidene, indicates CH. Further preferably, when the said aromatic heptylidene, caprylidene, oxymethylene, oxyethylidene, group is benzisoxazolyl, A indicates N. oxypropylidene, oxybutylidene, oxypentylidene, oxyhexy The said alicyclic groups preferably indicates five or six 50 lidene, oxyheptylidene, methyleneoxyl, ethylideneoxyl, pro membered saturated cycloalkyl groups, or five or six mem pylideneoxyl, butylideneoxyl, pentylideneoxyl, hexylide bered saturated heterocyclic groups with hetero atoms of N, neoxyl or heptylideneoxyl. Further more preferably, Y S. O. More preferably cyclopentyl, cyclohexyl, tetrahydrofu indicates methylene, ethylidene, propylidene, butylidene, ryl, piperidyl or piperazinyl groups. Further preferably cyclo pentylidene, hexylidene, heptylidene, caprylidene, methyl hexyl, piperidyl or piperazinyl groups. Particularly prefer 55 ably cyclohexyl group. eneoxyl, ethylideneoxyl, propylideneoxyl, butylideneoxyl, R indicates H. halogen, CN, C-C alkyl, C-C alkoxyl, pentylideneoxyl, hexylideneoxyl or heptylideneoxyl. Par CHO, CO(C-C alkyl), COO(C-C alkyl), COOH, NO, ticularly preferably, Y indicates ethylidene, propylidene, NH, NH(C-C alkyl), N(C-C alkyl), SH, S(C-C alkyl). butylidene, ethylideneoxyl or propylideneoxyl. Further par —S(O) (C-C alkyl). —S(O)H or —S(O)(C-C alkyl). 60 ticularly preferably, Y indicates propylidene, butylidene or alkyl parts in the above said groups are Substituted by anyone propylideneoxyl. Again particularly preferably, Y indicates or more halogen atoms. Preferably, R indicates H. halogen, propylidene or butylidene. Most preferably, when Yindicates CN, C-C alkyl, C-C alkoxyl, CHO, CO(C-C alkyl). butylidene, Xindicates N. Most preferably, when Yindicates COO(C-C alkyl) or COOH, alkyl parts in the above said propylidene, X indicates CH. groups are Substituted by anyone or more (e.g., 1-3 atoms) 65 In another embodiment, this invention offers the use of a halogen atoms. More preferably, R indicates H. F. Cl, Br, compound in formula (I) and its salts acceptable pharmaceu CN, C-C alkyl or C-C alkoxyl, CHO, COCH or tically in preparation of vasodilative drugs: US 9,415,047 B2 8 butylidene or propylideneoxyl. Again preferably, Y indicates (I) propylidene or butylidene. Most preferably, Y indicates pro pylidene. In another embodiment, this invention offers the use of a compound in formula (I) and its salts acceptable pharmaceu tically in preparation of vasodilative drugs:

(I) Wherein, 10 R indicates aromatic groups oralicyclic groups with mono or bisubstituted by R, wherein, The said aromatic groups indicate phenyl, benzisoxazolyl, benzofuranyl, benzisothiazolyl, benzopyranyl, benzoprya Zolyl or pyrimidinyl groups. The said alicyclic group indi 15 cates cyclohexyl. Preferably, if X indicates N, R indicates aromatic or ali cyclic groups mono or bisubstituted by R. and the said aro Wherein, matic groups indicate phenyl, benzisoxazolyl, benzofuranyl. R indicates aromatic groups mono orbisubstituted by R. benzisothiazolyl, benzopyranyl or pyrimidinyl groups. The and the said aromatic groups indicate phenyl, benzisoxazolyl, said alicyclic group indicates cyclohexyl. benzofuranyl, benzopryazolyl or benzisothiazolyl groups. Preferably, if X indicates CH, R indicates phenyl, ben Preferably, when X indicates N, the said aromatic group Zisoxazolyl, benzisothiazolyl, benzofuranyl or benzoprya indicates phenyl, benzisoxazolyl or benzisothiazolyl groups. Zolyl groups mono or bisubstituted by R. More preferably, 25 Further preferably, when X indicates CH, the said aromatic R indicates aromatic groups monosubstituted by R. and the group indicates phenyl, benzisoxazolyl, benzisothiazolyl, said aromatic groups preferably indicate phenyl or benzisoX benzofuranyl or benzopryazolyl group. azolyl. More preferably, when the said aromatic group is benzisoxazolyl, A indicates N. More preferably, when X indicates CH, R indicates aro R indicates H. F. Cl, Br, CN, C-C alkyl or C-C alkoxyl, 30 matic groups monosubstituted by R. and the said aromatic CHO, COCH, COOCH in which alkyl parts are substituted groups preferably indicate phenyl or benzisoxazolyl. Prefer 1-3 halogenatoms. Preferably, R indicates H.F, C1, CN, CF, ably, when the said aromatic group is benzisoxazolyl, A indi unsubstituted C-C alkyl or C-C alkoxyl, CHO, COCH or cates N. COOCH. More preferably, R indicates H, F, C1, CH, 35 R indicates H. F. Cl, Br, CN, C-C alkyl or C-C alkoxyl OCH COCH or CF. Further preferably, R indicates H, F, in which alkyl parts are substituted 1-3 halogen atoms. Pref Cl, CF, CN, CH or OCH. Particularly preferably, R indi erably, R indicates H. F. C1, CN, CF, unsubstituted C-C, cates H. F. Cl, CF, CH or OCH. Again preferably, R3 alkyl or C-C alkoxyl. More preferably, R indicates H. F. Cl, indicates H. For CF. Most preferably, R3 indicates F. If Rs. CF, CN, CH, or OCH. More preferably, R indicates H, F, indicates poly Substituted group, R is independently chosen 40 C1, CH, OCH or CF. Particularly preferably, R indicates from the above said groups. H. F or CF. Most preferably, R indicates F. If R indicates A, B and X respectively indicate CH or N. Preferably, A polysubstituted group, R is independently chosen from the and Ball indicate N. above said groups. R indicates H, F, Cl, Br, CN, CHO, COCH, COOCH or A, B and X respectively indicate CH or N. Preferably, A C-C alkyl or C-C alkoxyl in which alkyl parts are substi 45 and Ball indicate N. tuted 1-3 halogen atoms. Preferably, R indicates H. F. Cl, CN, CF, CH, OCH, CHO, COCH or COOCH. Further Rindicates H, F, Cl, Br, CN, CHO, COCH or C-C alkyl preferably, when Xindicates N, R indicates H. F. Clor CH. or C-C alkoxyl in which alkyl parts are substituted 1-3 Particularly preferably, when Xindicates CH, R indicates H. halogen atoms. More preferably, R indicates H. F. C1, CN, F C1, CN, CH, OCH or CHO, R more preferably indicates 50 CF, CH, OCH or COCH. Particularly preferably, R indi H or OCH R most preferably indicates H. If R indicates cates H, or OCH. Most preferably, R2 indicates H. poly Substituted group, R is independently chosen from the Y indicates Saturated or unsaturated Straight or branched above said groups. hydrocarbon chain composed of 2-8 carbon atoms, in which Y indicates Saturated or unsaturated Straight or branched 55 anyone or more carbonatoms are Substituted by hetero atoms hydrocarbon chain composed of 1-8 carbonatoms substituted Such as oxygen, Sulfur or nitrogen. Preferably, Y indicates by 2-3 halogenatoms, in which anyone or more carbonatoms unsubstituted Saturated alkyl group composed of 2-8 carbon are substituted by hetero atoms Such as oxygen, Sulfur or atoms, or unsubstituted Saturated alkyl group composed of nitrogen. Preferably, Yindicates unsubstituted saturated alkyl 2-8 carbon atoms in which one carbon atom is replaced by group composed of 2-8 carbon atoms, or unsubstituted Satu 60 rated alkyl group composed of 2-8 carbonatoms in which one oxygen or sulfur, e.g., —C, alkylidene-O-. More prefer carbon atom is replaced by oxygen or Sulfur, e.g., —C-7 ably, Y indicates ethylidene, propylidene, butylidene or eth alkylidene-O . More preferably, Y indicates ethylidene, ylideneoxyl. Particularly preferably, Y indicates propylidene propylidene, butylidene, pentylidene, hexylidene, ethylide or butylidene. Most preferably, Y indicates propylidene. neoxyl, propylideneoxyl or butylideneoxyl. Particularly pref 65 In another embodiment, this invention offers the use of a erably, Y indicates ethylidene, propylidene, butylidene or compound in formula (I) and its salts acceptable pharmaceu propylideneoxyl. Further preferably, Yindicates propylidene, tically in preparation of vasodilative drugs: US 9,415,047 B2 10

(I) (I)

10 Wherein, Wherein: when A and Ball indicateN, and Xindicates CH, R does not indicate benzoisoxazolyl substituted by 6-fluo R indicates aromatic groups mono orbisubstituted by R. rine wherein, R does not indicate H or Cl. And The said aromatic group indicates phenyl, benzisoxazolyl 15 Y dose not indicate ethylideneoxyl or propylideneoxyl. or benzisothiazolyl groups. Further preferably, when X indi In another embodiment, this invention offers the use of a cates N, the said aromatic group indicates phenyl or ben compound in formula (I) or its salts acceptable pharmaceuti Zisothiazolyl groups. Again preferably, when Xindicates CH, cally: the said aromatic group indicates phenyl or benzisoxazolyl group, more preferably benzisoxazolyl group. R indicates H. F. Cl, Br, OCH or CF. Particularly pref (I) erably, R indicates H. F. Clor CF. Further preferably, when X indicates N, R indicates H. C1 or CF. Again preferably, when X indicates CH, R indicates H. For CF. More pref erably, R3 indicates F. If R indicates polysubstituted group, 25 R is independently chosen from the above said groups. A, B and X respectively indicate CH or N. Preferably, A and Ball indicate N. Wherein: when A, B and X all indicate CH, R indicates H, F, Cl, Br, CN, CH or OCH. Particularly 30 R does not indicate benzoisoxazolyl substituted by 6-fluo preferably, R indicates H, or OCH. Most preferably, R2 rine indicates H. R does not indicate H, F, CN, COOCH or Cl. And Y indicates saturated straight or branched hydrocarbon Y dose not indicate ethylidene, propylidene, butylidene, pentylidene, ethylideneoxyl or propylideneoxyl. chain composed of 2-8 carbon atoms, in which anyone or 35 more carbonatoms are Substituted by hetero atoms of oxygen, In another embodiment, this invention offers the use of a Sulfur or nitrogen, e.g., —C, alkylidene-O-. Preferably, Y compound in formula (I) or its salts acceptable pharmaceuti indicates ethylidene, propylidene, butylidene or ethylide cally: neoxyl. More preferably, Y indicates propylidene or butyl idene. Further preferably, Y indicates butylidene, Xindicates 40 (I) N. Again preferably, if Y indicates propylidene, X indicates V-R2 CH. In another embodiment, this invention offers the use of a compound informula (I) and its salts acceptable pharmaceu ( s tically in preparation of vasodilative drugs: 45 R-X N-Y- \ / NN a (I) w 2 Wherein: when only A or B indicates CH, the other indi 50 cates N, and X indicates CH, R does not indicate benzoisoxazolyl substituted by 6-fluo ( s rine R-X N-Y- NN a R does not indicate H, For CN. And \ / Y does not indicate propylidene or butylidene. 55 In another embodiment, this invention offers the com Wherein, when A, B and X all indicate N. pound in formula (I) or its salts acceptable pharmaceutically R does not indicate phenyl group monosubstituted by Hor OCH and (I) R does not indicate H or OCH, OCH is substituted on 60 V-R2 benzo five-membered nitrogen heterocyclic ring. And Y dose not indicate ethylidene, propylidene, butylidene or ( s pentylidene. R-X N-Y- Ne In another embodiment, this invention offers the use of a 65 \ / B compound in formula (I) or its salts acceptable pharmaceuti cally: US 9,415,047 B2 11 12 Wherein, I-1 1-(4-(4-(3-chlorophenyl)piperazine-1-yl)butyl)-1H-ben R indicates phenyl or benzothiazol group monosubsti Zimidazole, tuted by R, wherein, I-2 1-(4-(4-(3-trifluoromethylphenyl)piperazine-1-yl)butyl)- R indicates H or Cl. If R indicates polysubstituted group, 1H-benzimidazole, R is independently chosen from the above said groups. I-3 1-(4-(4-(2,3-dichlorophenyl)piperazine-1-yl)butyl)-1H A and B independently indicate CH. benzimidazole, Xindicates N. I-4 1-(4-(4-(2-methoxyphenyl)piperazine-1-yl)butyl)-1H R indicates H or CN. benzimidazole, Y indicates butylidene. I-5 2-methyl-1-(4-(4-(3-trifluoromethylphenyl)piperazine In another embodiment, this invention offers the use of a 10 1-yl)butyl)-1H-benzimidazole, compound in formula (I) or its salts acceptable pharmaceuti I-6 6-fluoro-1-(4-(4-(3-trifluoromethylphenyl)piperazine-1- cally in preparation of vasodilative drugs: yl)butyl)-1H-benzimidazole, I-7 1-(3-(4-phenylpiperazine-1-yl)propyl)-1H-benzimida 15 Zole, (I) I-8 1-(3-(4-(3-fluorophenyl)piperazine-1-yl)propyl)-1H V-2 benzimidazole, I-9 2-methyl-1-(3-(4-(3-fluorophenyl)piperazine-1-yl)pro ( s pyl)-1H-benzimidazole, R-X N-Y- I-10 1-(4-(4-(3-cyanophenyl)piperazine-1-yl)butyl)-1H NN a benzimidazole, \ / I-11 1-(4-(4-(4-methylphenyl)piperazine-1-yl)butyl)-1H benzimidazole, Wherein, I-12 1-(4-(4-(2-furyl)piperazine-1-yl)butyl)-1H-benzimida R indicates benzisoxazolyl mono Substituted by R. 25 Zole, wherein R indicates H, F, Cl, Br, OCH or CF. Preferably, I-13 1-(4-(4-(4-pyridyl)piperazine-1-yl)butyl)-1H-benzimi R indicates H. For CF. More preferably, R indicates F. dazole, A, B and X respectively indicate CH or N. Preferably, A I-14 1-(4-(4-(2-pyrimidinyl)piperazine-1-yl)butyl)-1H-ben and Ball indicate N. X indicates CH. Zimidazole, R indicates H, OCH. Preferably, R2 indicates H. 30 I-15 1-(4-(4-(1-cyclohexyl)piperazine-1-yl)butyl)-1H-benz Y indicates saturated straight or branched hydrocarbon imidazole, chain composed of 2-8 carbon atoms, in which anyone or I-16 1-(4-(4-(1-naphthyl)piperazine-1-yl)butyl)-1H-benz more carbonatoms are Substituted by hetero atoms of oxygen, imidazole, Sulfur or nitrogen, e.g., —C, alkylidene-O-. Preferably I-171-(4-(4-(2-quinoxalinyl)piperazine-1-yl)butyl)-1H-ben 35 Zimidazole, ethylidene, propylidene or butylidene. More preferably, Y I-18 1-(4-(4-(3-(6-fluoro benzisoxazolyl))piperazine-1-yl) indicates propylidene or butylidene. Most preferably propy butyl)-1H-benzimidazole, lidene. I-19 1-(4-(4-(3-(6-fluoro benzisothiazolyl))piperazine-1-yl) In another embodiment, this invention offers the use of a butyl)-1H-benzimidazole, compound in formula (I) or its salts acceptable pharmaceuti 40 I-20 1-(4-(4-(3-benzimidazoyl)piperazine-1-yl)butyl)-1H cally in preparation of vasodilative drugs: benzimidazole, I-21 1-(4-(4-(3-(6-fluoro benzofuranyl))piperazine-1-yl)bu tyl)-1H-benzimidazole, (I) I-22 1-(3-(4-(3-(6-fluoro benzisoxazolyl))piperazine-1-yl) V 2 45 propoxyl)-1H-benzimidazole, I-23 1-(4-(4-(3-trifluoromethylphenyl)piperazine-1-yl)pro poxyl)-1H-benzimidazole, ( s I-24 1-(4-(4-(3-chlorphenyl)piperazine-1-yl)propoxyl)-1H R-X N-Y- NN a benzimidazole, \ / 50 I-25 6-chloro-1-(4-(4-(3-trifluoromethylphenyl)piperazine 1-yl)butyl)-1H-benzimidazole, Wherein, I-26 6-cyano-1-(4-(4-(3-trifluoromethylphenyl)piperazine R indicates benzisoxazolyl mono Substituted by R. 1-yl)butyl)-1H-benzimidazole, wherein R indicates H, F, Cl, Br or OCH. Preferably, R I-27 6-methoxycarbonyl-1-(4-(4-(3-trifluoromethylphenyl) indicates H or F. 55 piperazine-1-yl)butyl)-1H-benzimidazole, A and X indicate CH, B indicates CH or N. I-28 2-chloro-1-(5-(4-(3-trifluoromethylphenyl)piperazine R indicates H, F, Cl, Br, CN or OCH. 1-yl)pentyl)-1H-benzimidazole, Y indicates saturated straight or branched hydrocarbon I-29 1-(4-(4-(3-chlorophenyl)piperazine-1-yl)butyl)-1H chain composed of 2-8 carbon atoms, in which anyone or benzotriazole, more carbonatoms are Substituted by hetero atoms of oxygen, 60 I-30 1-(4-(4-(3-fluorophenyl)piperazine-1-yl)butyl)-1H Sulfur or nitrogen, e.g., —C, alkylidene-O-. Preferably benzotriazole, ethylidene, propylidene, butylidene, pentylidene, ethylide I-31 1-(4-(4-(3-trifluoromethylphenyl)piperazine-1-yl)bu neoxyl, propylideneoxyl or butylideneoxyl. Particularly pref tyl)-1H-benzotriazole, erably propylidene or butylidene. Most preferably propy I-32 6-fluoro-1-(4-(4-(3-trifluoromethylphenyl)piperazine lidene. 65 1-yl)butyl)-1H-benzotriazole, The said benzo five-membered nitrogen heterocyclic pip I-33 5,6-dimethyl-1-(4-(4-(3-trifluoromethylphenyl)pipera erazine or piperidine compounds include: Zine-1-yl)butyl)-1H-benzotriazole,

US 9,415,047 B2 17 18 -continued -continued

Code Chemical structure Code Chemical structure

C)s»©©OOCÓ?ý?#Š~ ,KOO\,, O)ZZZ*~,O CZ]()!}-\, US 9,415,047 B2 19 20 -continued -continued

Code Chemical structure e Chemical structure

I-18

I-26 I-19

I-27 25 I-20

I-21

I-28 O Cr- 35

I-22 I-29

I-23 I-30

I-24 US 9,415,047 B2 21 22 -continued -continued

Code Chemical structure e Chemical structure

I-32

I-40 I-33 15

I-41

I-34

I-42 30

I-35

35 I-43

I-36 I-44

50 I-45

I-37

I-46 I-38

US 9,415,047 B2 25 26 -continued -continued

Code Chemical structure Code Chemical structure

I-62 I-69

I-63 I-70 15

I-64

I-65 35 I-72

I-66

I-68 60

US 9,415,047 B2 29 30 -continued -continued

Code Chemical structure Code Chemical structure

I-88 5 I-94

N 1N N-N N

I-95

25 I-90 CH3

I-96 NC 30

35 OCH3

I-97 NC

I-92

55 I-98 OCH3 I-93 OCH

65 US 9,415,047 B2

-continued -continued

Code Chemical structure Code Chemical structure

I-99 5 /N N-ON I-105 C O N N N Nuo I 1 S N NS O N N I 10 NSN

I-100 HCO F I-106 C N S- \

NN N N Né 2O N N-N o1/ N o-N-NeN N F I-101 H3CO 25

N F HN1 N NN-N I-107 NeN 30 N C

N N F N-Nu Nea I-102 HCO 35

I-108

O N NN- C N N. N. 40 N-Nu! N N22 YN 2

F I-109 I-103 45 N N-N FC o1 N N N-Nu N N I Na NS 50 I-110 F FC I-104 NC 55 N-Nu N YN 2 N

o1NN N-Nu Na 60 I-111 N F N N N

65 US 9,415,047 B2 33 34 -continued -continued

Code Chemical structure Code Chemical structure

I-112 5 I-119

OCH N-Nu

I-120 I-113 F 2) 15 CyN Cr-C N 20 I-121

I-114 H3CO - N RN 25 Né

FC N-Nu I-122

I-115 NC N 35 I-123

FC N N N n

N 40 2

FC 45 C 3 N-N -N N N-Nu N N

I-117 I-125 C () FC

3 N-N e N N 55 N-Nu NS 2N O Na N

I-126 I-118 60

Na US 9,415,047 B2

-continued -continued

Code Chemical structure Code Chemical structure

I-127 I-134

FC FC N-Nu N N?'N 10 3 N-N O-NeN

I-128 I-135 N F 15 / NN N-N N-Nu N Na N O Ns

2O I-129 HC OCH N I-136 NNa N 25 o1 NN NN-N N

NSN I-130 F 30

HCO FC N-Nu NN 2N 35 I-137 N F N O 1. NN N-N N

I-131 HCO Ns 40

F FC 3 N-Nu NQ 2N 45 I-138 C N N o1 N N-N N I-132 NC I 50 NSNN

FC 3 N-Nu F NeNN 55 I-139 CH3

N I-133 o1 N NN-N

65 F

US 9,415,047 B2 46

II-34 II-2

FC -N NN- N-Nu NaN HCI 10

Hydrochlorate is preferred for salt of compound I-3, i.e., compound II-3. The corresponding chemical structure is In an embodiment of this invention, especially the prefer illustrated as II-3. 15 able compound I-85 and its pharmaceutically acceptable salts, e.g., hydrochlorate, i.e., compound II-85. The corre sponding chemical structure is illustrated as follows.

II-3 2O

C N N N-N C -\ o1 N N NN- N-Nu NeN 25 HCI s HCI

F Of which compound I-29 is a preferable hydrochlorate, 30 i.e., compound II-29. The corresponding chemical structure In vitro animal experiment Suggested that, compound II-2 is illustrated as II-3. could relax vascular Smooth muscles constricted by adrena line and high potassium concentration, with -log ECso values of relaxation of 5.73+0.03 and 5.34+0.02, respectively (FIG. 35 1 and FIG. 2). Compound II-3 could relax vascular smooth II-29 muscles constricted by adrenaline and high potassium con centration, with -log ECso values of relaxation of 6.01 +0.05 and 5.49+0.05, respectively (FIG. 8 and FIG.9). 40 It was indicated in a study on dilating mechanism of com pound II-2 on vascular Smooth muscle that, the compound C -N could competitively resist the vasoconstrictive effects of Nu N-Nu N.N? 2 N. noradrenaline, calcium ions and hydroxytryptamine, move HCI the dose effect curve of above said agonist transversally to 45 right, while maximal response was not reduced, suggesting that PA values for the compound to resist vasoconstrictive effects of noradrenaline (NA), calcium ions and hydrox Hydrochlorate is preferred for salt of compound I-31, i.e., ytryptamine (5-HT) were 7.37+0.08 (7.52+0.04 for dox compound II-31. The corresponding chemical structure is azosin), 5.61+0.04 (6.99+0.05 for amlodipine) and 5.71+0.08 50 (FIGS. 3, 4, 5, 6 and 7). The results indicated that, compound illustrated as II-31. II-2 produced vasodilative effects by blocking C. receptor, Ca" ion channel and vascular 5-HT, receptor. In vitro animal experiment Suggested that, compound II-29 could relax vascular Smooth muscles constricted by adrena II-31 55 line and high potassium concentration, with -log ECso values of relaxation of 6.01+0.02 and 5.64+0.01, respectively (FIG. 15 and FIG. 16). Compound II-31 could relax vascular Smooth muscles constricted by adrenaline and high potas sium concentration, with -log ECso values of relaxation of FC N -NN-Nu 60 6.19+0.03 and 5.55+0.03, respectively (FIG.10 and FIG.11). N- NaN It was indicated in a study on dilating mechanism of com pound II-31 on vascular Smooth muscle that, the compound could competitively resist the vasoconstrictive effects of noradrenaline, calcium ions and hydroxytryptamine, move Hydrochlorate is preferred for salt of compound I-34, i.e., 65 the dose effect curve of above said agonist transversally to compound II-34. The corresponding chemical structure is right, while maximal response was not reduced, suggesting illustrated as II-34. that PA values for the compound to resist vasoconstrictive US 9,415,047 B2 47 48 effects of noradrenaline, calcium ions and hydrox hypofunction resulted by renal vasospasm and diseases ytryptamine were 6.02+0.13 (7.76+0.24 for doxazosin), related with peripheral vascular spasm. 6.56+0.032 (7.51+0.288 for amlodipine) and 6.726+0.089 Subjects described in this invention should be mammals (FIGS. 12, 13 and 14). These results indicated that, compound preferably, especially human. II-31 produced vasodilative effects by blocking C. receptor, This invention offers compound in formula (I) or its salts Ca" ion channel and vascular 5-HT, receptor. acceptable pharmaceutically preferably for prevention, alle In in Vivo bulk testing on rats, compound II-2 showed good viation or treatment against hypertension, angina pectoris, hypotensive effects, good oral absorption, mild toxicity, great heart failure, coronary heart disease, cerebral ischemia and therapeutic index, negative marrow micronucleus test, with peripheral vascular spasmodic diseases, such as thromboan protential value in development of multiple target vasodila 10 gitis obliterans and raynauds disease, etc. tive drugs. Compound in formula (I) and its salts acceptable pharma The vivo bulk testing on hypotensive effects in rats indi cological in this invention can be prepared into appropriate cated that, compound II-85 had obvious hypotensive effects, complexes for oral, parenteral, nasal spraying, rectal, intra good oral absorption, relatively mild acute toxicity, greater nasal, Sublingual, intrabuccal, percutaneous or implantable therapeutic index, negative marrow micronucleus test, with 15 administrations, and the said parenteral administration protential values in development of vasodilative drugs, espe includes Subcutaneous, intradermal, intravneous, intramus cially new hypotensive drugs. cular, intraarticular, intrasynovial, intrasternal, intrathecal, The inventor found that, the said compound in formula (I) focal, intracranial or instillation techniques. Compound and and its salts acceptably pharmaceutically in this invention its salts acceptable pharmaceutically in this invention (I) are have obvious relaxing effects on vascular Smooth muscles of preferred to be administered by oral, Sublingual, intravenous, Subjects. The said compound in formula (I) and its salts intramuscular and Subcutaneous approaches. The said drug acceptable pharmaceutically in this invention may produce complexes may contain one or more routine medicinal car relaxing effects on vascular Smooth muscles by antagonism rier, adjuvant or media, e.g., diluent, excipient like water, etc.; against a receptors (especially C. receptors). In addition, adhesives like fibrin derivatives, gelatin, poly Vidone, etc.; compound in formula (I) and its salts acceptably pharmaceu 25 bulking agent like starch, etc.; disintegrant like calcium car tically in this invention may achieve relaxing effects on vas bonate, sodium bicarbonate, etc.; lubricant like calcium Stear cular Smooth muscles by acting on other targets or ate or magnesium Stearate, etc.; and other adjuvants like fla approaches, e.g., Ca" channel blockade or antagonism Vor and Sweetener. against 5-HT2 receptors. Compounds with multiple targets The said drug complexes containing compound informula in this invention are particularly preferable, e.g., compound 30 (I) and its salts acceptable pharmaceutically in this invention I-2 or II-2, compound I-85 or II-85. With multiple targets, may be in the form of sterile injection Solution, e.g., sterile compounds in this invention are particularly effective against aqueous or oily suspension. This suspension may be prepared diseases related with persistent and pathological vascular by using appropriate dispersing agent or lubricant (e.g., constriction or spasm of vascular Smooth muscle. When the Tween 80) and Suspending agent based on known techniques compounds are used in combination with one or more single 35 in this field. Sterile injection solution may also be sterile target drugs against the said diseases, higher efficacy may be injection solution or Suspension applicable to nontoxic dilu achieved, or drug resistance or undesireble side effects may ent or solvent for parenteral medications, e.g., Solution in be effectively reduced, thus improving safety. Specifically, 1,3-butylene glycol. Applicable media and solvents may be because of multiple target effects, when C. receptor of vas mannitol, water, Ringer's Solution and isotonic sodium chlo cular Smooth muscle is incompletely blocked, good hypoten 40 ride solution. In addition, sterile nonvolatile oil may be rou sive efficacy may be produced by Synergistically blocking tinely used as solvent or Suspension media. Therefore, any Ca" channel and/or 5-HT, receptor, then the remaining C. mild nonvolatile oil may be used, including synthetic receptors could still participate in pressor reflex, which may monoglyceride and diacylglycerol. Fatty acids, e.g., oleinic prevent and reduce the occurrence of orthostatic hypotension. acid and its glyceride derivatives, may be used in injection And/or by blocking Ca" channel, in addition to synergistic 45 preparation, or natural medicinal oil may also be used, e.g., hypotensive effects, effects may be produced to resist myo olive oil or castor oil, especially its polyoxyethylene com cardial hypertrophy, protect vascular endothelia, resist ath pound. The said oily solution or Suspension also includes long erosclerosis, inhibit hyperplasia of vascular Smooth muscle, chain alcohol or dispersing agent (including those described and improve cerebral blood circulation, and prevent occur in Ph. Helv) or similar alcohol. rence of first dose effect by reducing heart rate, effectively 50 The said drug complexes containing compound informula preventing tachycardia and palpitation. And/or by blocking (I) and its salts acceptable pharmaceutically in this invention 5-HT2 receptors, in addition to synergistic hypotensive may be orally administered in any oral dosage form, and the effects, the drug combination may effectively improve blood said dosage forms include, but not limit to, capsules, tablets, Supply to patients with occlusion vascular diseases, then the powders, granules, aqueous Suspension and solution. The compound may be used to hypertension patients with athero 55 said dosage form is prepared by using mature techniques in Sclerosis and endothelial injuries. the field of pharmaceutical preparation. For oral tablets, car Therefore, compound in formula (I) and the salts accept riers usually used include lactose and corn starch. Lubricant is able pharmaceutically in this invention may be used to pre usually added (e.g., magnesium Stearate). For oral adminis vent, alleviate or treat Subjects with diseases or symptoms tration in the form of capsules, the applicable diluents include related with persistent and pathological constriction or vas 60 lactose and dry corn starch. When aqueous Suspension is cular spasm. The said compound in formula (I) and the salts orally administered, active components can bind to emulsi acceptable pharmaceutically may be specially used to pre fying and Suspending agents. If necessary, Some Sweeteners vent, alleviate or treat hypertension, heart failure, angina pec and/or flavors and/or colorants can be added. toris and coronary heart diseases, etc. The compound and its The said drug complexes containing compound informula salts may be used to treat cerebral ischemic diseases, myo 65 (I) and its salts acceptable pharmaceutically in this invention cardial ischemic diseases, shock, etc. induced by vascular may be used in the form of nasal aerosol or inhaled. These spasm. The compound and its salts may be used to treat renal complexes can be prepared by using mature techniques in the US 9,415,047 B2 49 50 field of pharmaceutical preparation into Saline Solutions by using benzoic alcohol or other appropriate antiseptics, absor -continued FVR befacient to improve bioavailability, fluorocarbon and/or -NN 1Nu-N \ 2 M other known solubilizing agents or dispersants in this field. X -e- The said drug complexes containing compound informula R- N- s (I) and its salts acceptable pharmaceutically in this invention may be used in the form of rectal Suppository. The said com (I) plexes are prepared by mixing compound in this invention and appropriate non-irritant excipient, and the said excipient is solid at ambient temperature but liquid at rectal tempera 10 ture, and the complex will release active components after dissolution in rectum. These substance include but not limit to, cacao butter, bees wax and polyethylene glycol. (II) As deduced based on results of rat experiment, the daily 15 dose of compound in formula (I) in this invention should be Wherein, R. R. A. Band Xare described in the above text. less than the daily dose of amlodipine. In this field, the daily n=0-7. M indicates medical drugs, e.g., HCl, 2HCl, HBr, dose is known for amlodipine to relax blood vessels or treat 2HBr, HSO CHSOH, etc. hypotension, e.g., 10 mg/day. The specific dose of this com In sodium hydroxide solution, substituted 1H-benzo five pound in formula (I) in this invention may be determined membered nitrogen heterocyclic ring is used as raw material based on results of clinical trial, patient’s conditions and age, to condense with chloro-alkyl bromide to prepare N-chloro etc. alkyl substituted benzo five-membered nitrogen heterocyclic The said drug complexes containing compound informula compound, and condense with Substituted piperazine and (I) and its salts acceptable pharmaceutically in this invention piperidine to prepare the indicated compound in formula (I), may be prepared by using routine method in medical field, 25 finally corresponding salt will be prepared by acidification to with 0.1%-99.5% w/w of active ingredients, which may be produce compound informula (II). The above method may be determined by diseases to be treated or prevented, and con used to prepare compound I-1 to I-21, I-25 to I-51, I-54. to ditions of Subjects to whom the said compound may be I-60, I-84 to I-87, I-100 to I-102, I-124 to I-132, I-135 to administered. Dosage regimen of the administered com I-146, I-149 to I-157, and their salts. pound can be easily determined by technicians in the field 30 Synthetic Approach 2: based on contents publicized in this document. In another embodiment, the compound informula (I) or its salts acceptable pharmaceutically in this invention may be OH combined with one or more other active pharmaceutical R2 ingredients. This drug combination may be a single complex 35 N 2/ NaH containing compound or its salts acceptable pharmaceutically A. C in this invention and one or more other active ingredients, or \, S N-1NB combination of two or more complexes, where compound in this invention is contained in one complex, while one or more 40 C / y R2 R-N NH other active ingredients are contained in one or more separate complexes. Compound in formula (I) or the salts acceptable N-r W \ / pharmaceutically in this invention may be combined with As o FVR other active ingredients, such as antispasmodic against O X Smooth muscle spasm, preferably Sertraline, captopril, 45 benazepril, Valsartan, inderal and diuretics, to prevent, alle ?\-hu- NN \ / M viate or treat subjects with diseases or symptoms related with R-\u? s persistent and pathological constriction or vascular spasm. Except otherwise specified, the embodiments described in (I) this application, or regimens with different preferabilities, 50 O = y.R. may be freely combined. Compound in this invention can be synthesized by adopt X ing the following methods: R- N- M As Synthetic Approach 1: 55 (II)

H R2 N 2y. NaOHIHO Wherein, R, R, A, B and Xare described in the above text. A. C n=0-7. M indicates medical drugs, e.g., HCl, 2HCl, HBr, \, N pi Br 60 2HBr, HSO CHSOH, etc. Substituted benzo five-membered nitrogen heterocyclic-1- ol is used as raw material to exchange active hydrogen with / yW R2 R -N NH Sodium hydride to produce corresponding salt, and react with chloro-alkyl bromide to produce corresponding chloride, and 65 condense with piperazine and piperidine to prepare the indi cated compound informula (I), finally corresponding salt will be prepared by acidification to produce compound informula US 9,415,047 B2 51 52 (II). The above method may be used to prepare compound Anhydrous magnesium Sulfate is added to dry organic phase, I-22 to I-24, I-52 to I-53, I-98 to I-99, I-133 to I-134, I-147 to filtered, with solvent evaporated. Oily products are analyzed I-148, and their salts. by chromatography with neutral Al2O, or separated and puri Common method one for synthesis: preparation of N-(4- fied by using HPLC to prepare 1-(2-chloroethoxyl)-substi chlorobutyl)-substituted benzo five-membered nitrogen het tuted benzo five-membered nitrogen heterocyclic compound, erocyclic compound with a yield range of 75.0%-85.0%. 1H-substituted benzo five-membered nitrogen heterocy The following examples are combined to illustrate this clic compound (0.10 mol) is dissolved into 100 ml of 30% wt. invention. sodium hydroxide, 4-chlorobromobutane (34.0 g, 0.10 mol), tetrabutyl ammonium bromide (0.8 g) are added, and mixed 10 Example 1 for 5 min. The reaction solution is gradually heated to 60°C., stirred for reaction for 2 hours. Then the reaction solution is Preparation of 1-(4-(4-(3-chlorophenyl)piperazine-1- cooled down to ambient temperature, 100 ml of dichlo yl)butyl)-1H-benzimidazole (I-1) romethane is added for extraction and liquid separation. To the aqueous phase, 100 of dichloromethane is added for 15 extraction. Organic phases are mixed, washed with 100 ml of 1H-benzimidazole (11.8 g., 0.10 mol) was dissolved into saturated Saline. Liquid is separated, and organic phase is 200 ml of 20% wt. sodium hydroxide, 4-chlorobromobutane evaporated to dryness to produce oily product. Oily products (34.3 g, 0.20 mol) and tetrabutyl ammonium bromide (1.0 g) are analyzed by chromatography with neutral Al2O, or sepa were added, and mixed for 5 min. The mixture was heated to rated and purified by using HPLC to prepare N-(4-chlorobu 60° C., stirred to react for 2 hours. Then the reaction solution tyl)-substituted benzo five-membered nitrogen heterocyclic was cooled down to ambient temperature, 100 ml of dichlo compound, with a yield range of 30.0%-85.0%. romethane was added for extraction and liquid separation. To Common method two for synthesis: preparation of N-(3- the aqueous phase, 100 of dichloromethane was added for chloropropyl)-substituted benzo five-membered nitrogen extraction. Organic phases were mixed, washed with 100 ml heterocyclic compound 25 of Saturated Saline. Liquid was separated, and organic phase 1H-substituted benzo five-membered nitrogen heterocy was evaporated to dryness to produce oily product. Oily prod clic compound (0.10 mol) is dissolved into 100 ml of 30% wt. ucts were separated and purified by chromatography with sodium hydroxide, 3-chlorobromopropane (31.4 g 0.10 neutral Al-O to produce 12.5 g of 1-(4-chlorobutyl)-1H mol), tetrabutyl ammonium bromide (0.8 g) are added, and benzimidazole, with a yield of 60.0%. mixed for 5 min. The reaction solution is gradually heated to 30 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) 60° C., stirred for reaction for 2 hours. Then the reaction was dissolved into 100 ml of acetonitrile, 3-trichlorophe solution is cooled down to ambient temperature, 100 ml of nylpiperazine (5.9 g, 0.03 mol), diisopropylethylamine (15.5 dichloromethane is added for extraction and liquid separa g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were tion. To the aqueous phase, 100 of dichloromethane is added respectively added. The mixture was stirred for 10 min at for extraction. Organic phases are mixed, washed with 100 ml 35 ambient temperature, and then heated and refluxed to react for of Saturated Saline. Liquid is separated, and organic phase is 15 hours. The mixture was cooled down to ambient tempera evaporated to dryness to produce oily product. Oily products ture and filtered. The filtrate was concentrated to produce oily are analyzed by chromatography with neutral Al2O, or sepa products, and treated by chromatography with neutral Al-O. rated and purified by using HPLC to prepare N-(3-chloropro eluted with dichloromethane/methanol mixture to produce pyl)-substituted benzo five-membered nitrogen heterocyclic 40 6.8g compound (I-1) with a yield of 61.4%. ESI-MSM--H": compound, with a yield range of 30.0%-85.0%. m/Z 369.2. Common method three for synthesis: preparation of N-(3- substituted benzo five-membered nitrogen heterocyclic) Example 2 proply-4-Substituted piperidine N-(3-chloropropyl)-substituted benzo five-membered 45 Preparation of 1-(4-(4-(3-trifluoromethylphenyl) nitrogen heterocyclic compound (0.06 mol) into 150 ml of piperazine-1-yl)butyl)-1H-benzimidazole (I-2) and acetonitrile, 4-substituted piperidine (0.05 mol), diisopropy 1-(4-(4-(3-trifluoromethylphenyl)piperazine-1-yl) lethylamine (0.2 mol) and potassium iodide (0.05 mol) are butyl)-1H-benzimidazole hydrochlorate (II-2) respectively added. The mixture is mixed at ambient tempera ture for 10 min, then heated and refluxed for reaction for 50 The method described in Example 1 was adopted to pre 10-20hours. The mixture is cooled downto ambient tempera pare 1-(4-chlorobutyl)-1H-benzimidazole. ture and filtered. The filtrate is concentrated to produce oily 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) products, and treated by chromatography with neutral Al2O, was dissolved into 100 ml of acetonitrile, 3-trichloromethyl eluted with dichloromethane/methanol mixture to produce phenylpiperazine (6.91 g, 0.03 mol), diisopropylethylamine compound in formula (I) with a yield range of 65.0%-72.0%. 55 (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) Common method four for synthesis: preparation of N-(2- were respectively added. The mixture was stirred for 10 min chloroethoxyl)-substituted benzo five-membered nitrogen at ambient temperature, and then heated and refluxed to react heterocyclic compound for -20 hours. The mixture was cooled down to ambient Substituted N-hydroxyl benzo five-membered nitrogen temperature and filtered. The filtrate was concentrated to heterocyclic compound (0.01 mol) is dissolved in 10 ml of 60 produce oily products, and treated by chromatography with NMP solid paraffin mixture containing 50% (w/w) hydrogen neutral Al-O, eluted with dichloromethane/methanol mix and oxygen is added in different times, stirred to react for 0.5 ture to produce 7.6 g compound (I-2) with a yield of 62.8%. h. Meanwhile, 3-bromochloropropane (0.015 mol) is dis Compound (I-2) (6.04g, 0.015 mol) was dissolved in 80 ml solved in 5 ml of NMP and added into the above said solution, of ethyl acetate and 8 ml of ethanol. Under cooling conditions and stirred to react for 12 h. Reaction solution is poured into 65 of icy water bath, 3 mol/L hydrogen chloride/ethyl acetate 50 ml of water, extracted with ethyl acetate (3x50 mL). solution is dripped, and the pH value is adjusted to 3. The Organic phases are mixed and washed with 30 ml of water. mixture is heated to 50° C. and stirred for 20 min, cooled US 9,415,047 B2 53 54 down for recrystallization, filtered and dried to produce 5.9 g, romethane was added for extraction. Organic phases were solid compound (II-2) with a yield of 89.7%. ESI-MS mixed, washed with 100 ml of saturated saline. Liquid was M+H": m/z 403.2. separated, and organic phase was evaporated to dryness to produce oily product. Oily products were separated and puri Example 3 fied by chromatography with neutral Al-O to produce 13.7g of 1-(4-chlorobutyl)-2-methyl-1H-benzimidazole, with a Preparation of 1-(4-(4-(2,3-dichlorophenyl)pipera yield of 61.5%. Zine-1-yl)butyl)-1H-benzimidazole (I-3) and 1-(4-(4- 1-(4-chlorobutyl)-2-methyl-1H-benzimidazole (8.02 g, (2,3-dichlorophenyl)piperazine-1-yl)butyl)-1H-benz 0.036 mol) was dissolved into 100 ml of acetonitrile, imidazole hydrochlorate (II-3) 10 3-trichloromethyl phenylpiperazine (6.91 g, 0.03 mol), diiso propylethylamine (15.5 g., 0.12 mol) and potassium iodide The method described in Example 1 was adopted to pre (5.0 g, 0.03 mol) were respectively added. The mixture was pare 1-(4-chlorobutyl)-1H-benzimidazole. stirred for 10 minatambient temperature, and then heated and 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) refluxed to react for 15 hours. The mixture was cooled down was dissolved into 100 ml of acetonitrile, 2,3-dicholorophe 15 nylpiperazine (6.93 g, 0.03 mol), diisopropylethylamine to ambient temperature and filtered. The filtrate was concen (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) trated to produce oily products, and treated by chromatogra were respectively added. The mixture was stirred for 10 min phy with neutral Al-O, eluted with dichloromethane/metha at ambient temperature, and then heated and refluxed to react nol mixture to produce 8.1 g compound (I-5) with a yield of for 10-20 hours. The mixture was cooled down to ambient 64.9%. ESI-MSM--H: m/z 417.2. temperature and filtered. The filtrate was concentrated to produce oily products, and treated by chromatography with Example 6 neutral Al-O, eluted with dichloromethane/methanol mix ture to produce 7.5 g compound (I-3) with a yield of 62.0%. Preparation of 6-fluoro-1-(4-(4-(3-trifluorometh Compound (I-3) (6.05 g, 0.015 mol) was dissolved in 80 ml 25 ylphenyl)piperazine-1-yl)butyl)-1H-benzimidazole of ethyl acetate and 8 ml of ethanol. Under cooling conditions (I-6) of icy water bath, 3 mol/L hydrogen chloride/ethyl acetate solution is dripped, and the pH value is adjusted to 3. The 6-fluoro-1H-benzimidazole (13.2 g, 0.10 mol) was dis mixture is heated to 50° C. and stirred for 20 min, cooled solved into 200 ml of 20% wt. sodium hydroxide, 4-chloro down for recrystallization, filtered and dried to produce 6.0 g 30 bromobutane (34.3 g, 0.20 mol) and tetrabutyl ammonium solid compound (II-3) with a yield of 90.9%. ESI-MS bromide (1.0 g) were added, and mixed for 5 min. The mix M+H": m/z 403.1. ture was heated to 60° C., stirred to react for 2 hours. Post treatment was performed based on common method one for Example 4 synthesis. Oily products were separated and purified by chro 35 matography with neutral Al-O to produce 14.2 g of 1-(4- Preparation of 1-(4-(4-(2-methoxyphenyl)piperazine chlorobutyl)-6-fluoro-1H-benzimidazole, with a yield of 1-yl)butyl)-1H-benzimidazole (I-4) 62.6% 1-(4-chlorobutyl)-6-fluoro-1H-benzimidazole (8.16 g. The method described in Example 1 was adopted to pre 0.036 mol) was dissolved into 100 ml of acetonitrile, pare 1-(4-chlorobutyl)-1H-benzimidazole. 40 3-trichloromethyl phenylpiperazine (6.91 g, 0.03 mol), diiso 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) propylethylamine (15.5 g., 0.12 mol) and potassium iodide was dissolved into 100 ml of acetonitrile, 2-methoxyphenyl (5.0 g, 0.03 mol) were respectively added. The mixture was piperazine (5.77g, 0.03 mol), diisopropylethylamine (15.5 g. stirred for 10 minatambient temperature, and then heated and 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec refluxed to react for 15 hours. The mixture was cooled down tively added. The mixture was stirred for 10 min at ambient 45 to ambient temperature and filtered. The filtrate was concen temperature, and then heated and refluxed to react for 10-15 trated to produce oily products, and treated by chromatogra hours. The mixture was cooled down to ambient temperature phy with neutral Al-O, eluted with dichloromethane/metha and filtered. The filtrate was concentrated to produce oily nol mixture to produce 8.5 g compound (I-6) with a yield of products, and treated by chromatography with neutral Al2O, 67.4%. ESI-MSM--H: m/z 421.2. eluted with dichloromethane/methanol mixture to produce 50 7.7 g compound (I-4) with a yield of 70.6%. ESI-MSM--HI": Example 7 m/Z 365.2. Preparation of 1-(3-(4-phenylpiperazine-1-yl)pro Example 5 pyl)-1H-benzimidazole (I-7) 55 Preparation of 2-methyl-1-(4-(4-(3-trifluorometh 1H-benzimidazole (11.8 g., 0.10 mol) was dissolved into ylphenyl)piperazine-1-yl)butyl)-1H-benzimidazole 200 ml of 20% wt. sodium hydroxide, 3-chlorobromopropane (I-5) (31.4g, 0.20 mol) and tetrabutyl ammonium bromide (1.0 g) were added, and mixed for 5 min. The mixture was heated to 2-methyl-1H-benzimidazole (13.2 g, 0.10 mol) was dis 60 60° C., stirred to react for 2 hours. Then the reaction solution solved into 200 ml of 20% wt. sodium hydroxide, 4-chloro was cooled down to ambient temperature, 100 ml of dichlo bromobutane (34.3 g, 0.20 mol) and tetrabutyl ammonium romethane was added for extraction and liquid separation. To bromide (1.0 g) were added, and mixed for 5 min. The mix the aqueous phase, 100 of dichloromethane was added for ture was heated to 60°C., stirred to react for 2 hours. Then the extraction. Organic phases were mixed, washed with 100 ml reaction solution was cooled down to ambient temperature, 65 of Saturated Saline. Liquid was separated, and organic phase 100 ml of dichloromethane was added for extraction and was evaporated to dryness to produce oily product. Oily prod liquid separation. To the aqueous phase, 100 of dichlo ucts were separated and purified by chromatography with US 9,415,047 B2 55 56 neutral Al-O to produce 12.0 g of 1-(3-chloropropyl)-1H Example 10 benzimidazole, with a yield of 62.0%. 1-(3-chloropropyl)-1H-benzimidazole (6.98 g., 0.036 mol) Preparation of 1-(4-(4-(3-cyanophenyl)piperazine-1- was dissolved into 100 ml of acetonitrile, phenyl piperazine yl)butyl)-1H-benzimidazole (I-10) (4.9 g, 0.03 mol), diisopropylethylamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were respectively The method described in Example 1 was adopted to pre added. The mixture was stirred for 10 min at ambient tem pare 1-(4-chlorobutyl)-1H-benzimidazole. perature, and then heated and refluxed to react for 10-15 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) hours. The mixture was cooled down to ambient temperature was dissolved into 100 ml of acetonitrile, 3-cyanophenyl and filtered. The filtrate was concentrated to produce oily 10 piperazine (5.6 g., 0.03 mol), diisopropylethylamine (15.5 g. products, and treated by chromatography with neutral Al2O, 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec eluted with dichloromethane/methanol mixture to produce tively added. The mixture was stirred for 10 min at ambient 6.1 g compound (I-7) with a yield of 63.2%. ESI-MSM--HI": temperature, and then heated and refluxed to react for 10-15 m/Z 321.2. hours. The mixture was cooled down to ambient temperature 15 and filtered. The filtrate was concentrated to produce oily Example 8 products, and treated by chromatography with neutral Al2O, eluted with dichloromethane/methanol mixture to produce Preparation of 1-(3-(4-(3-fluorophenyl)piperazine-1- 6.7 g compound (I-10) with a yield of 62.4%. ESI-MS yl)propyl)-1H-benzimidazole (I-8) M+H: m/z. 360.2. The method described in Example 7 was adopted to pre Example 11 pare 1-(3-chloropropyl)-1H-benzimidazole. 1-(3-chloropropyl)-1H-benzimidazole (6.98 g., 0.036 mol) Preparation of 1-(4-(4-(4-methylphenyl)piperazine was dissolved into 100 ml of acetonitrile, 3-fluorophenyl 1-yl)butyl)-1H-benzimidazole (I-11) piperazine (6.91 g, 0.03 mol), diisopropylethylamine (15.5 g. 25 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec The method described in Example 1 was adopted to pre tively added. The mixture was stirred for 10 min at ambient pare 1-(4-chlorobutyl)-1H-benzimidazole. temperature, and then heated and refluxed to react for 10-15 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) hours. The mixture was cooled down to ambient temperature was dissolved into 100 ml of acetonitrile, 4-methylphenyl and filtered. The filtrate was concentrated to produce oily 30 piperazine (5.3.g., 0.03 mol), diisopropylethylamine (15.5 g. products, and treated by chromatography with neutral Al-O. 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec eluted with dichloromethane/methanol mixture to produce tively added. The mixture was stirred for 10 min at ambient 6.4g compound (I-8) with a yield of 63.1%. ESI-MSM--H": temperature, and then heated and refluxed to react for 10-15 m/Z 339.2. hours. The mixture was cooled down to ambient temperature 35 and filtered. The filtrate was concentrated to produce oily Example 9 products, and treated by chromatography with neutral Al-O. eluted with dichloromethane/methanol mixture to produce Preparation of 2-methyl-1-(3-(4-(3-fluorophenyl) 6.4 g compound (I-11) with a yield of 60.7%. ESI-MS piperazine-1-yl)propyl)-1H-benzimidazole (I-9) M+H: m/z 349.2. 40 2-methyl-1H-benzimidazole (13.2 g, 0.10 mol) was dis Example 12 solved into 200 ml of 20% wt. sodium hydroxide, 3-chloro bromopropane (31.4g, 0.20 mol) and tetrabutyl ammonium Preparation of 1-(4-(4-(2-furyl)piperazine-1-yl)bu bromide (1.0 g) were added, and mixed for 5 min. The mix tyl)-1H-benzimidazole (I-12) ture was heated to 60°C., stirred to react for 2 hours. Then the 45 reaction solution was cooled down to ambient temperature, The method described in Example 1 was adopted to pre 100 ml of dichloromethane was added for extraction and pare 1-(4-chlorobutyl)-1H-benzimidazole. liquid separation. To the aqueous phase, 100 of dichlo 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) romethane was added for extraction. Organic phases were was dissolved into 100 ml of acetonitrile, 4-(2-furyl) pipera mixed, washed with 100 ml of saturated saline. Liquid was 50 zine (4.6 g., 0.03 mol), diisopropylethylamine (15.5 g., 0.12 separated, and organic phase was evaporated to dryness to mol) and potassium iodide (5.0g, 0.03 mol) were respectively produce oily product. Oily products were separated and puri added. The mixture was stirred for 10 min at ambient tem fied by chromatography with neutral Al-O to produce 12.9 g, perature, and then heated and refluxed to react for 20 hours. of 1-(3-chloropropyl)-2-methyl-1H-benzimidazole, with a The mixture was cooled down to ambient temperature and yield of 62.1%. 55 filtered. The filtrate was concentrated to produce oily prod 1-(3-chloropropyl)-2-methyl-1H-benzimidazole (7.49 g, ucts, and treated by chromatography with neutral Al2O, 0.036 mol) was dissolved into 100 ml of acetonitrile, 3-tri eluted with dichloromethane/methanol mixture to produce cholo phenylpiperazine (4.9 g, 0.03 mol), diisopropylethy 6.0 g compound (I-12) with a yield of 61.5%. ESI-MS lamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 M+H: m/z 325.2. mol) were respectively added. The mixture was stirred for 10 60 min at ambient temperature, and then heated and refluxed to Example 13 react for 15 hours. The mixture was cooled down to ambient temperature and filtered. The filtrate was concentrated to Preparation of 1-(4-(4-(4-pyridyl)piperazine-1-yl) produce oily products, and treated by chromatography with butyl)-1H-benzimidazole (I-13) neutral Al-O, eluted with dichloromethane/methanol mix 65 ture to produce 6.67 g compound (I-9) with a yield of 63.1%. The method described in Example 1 was adopted to pre ESI-MSM--H": m/z. 353.2. pare 1-(4-chlorobutyl)-1H-benzimidazole. US 9,415,047 B2 57 58 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) filtered. The filtrate was concentrated to produce oily prod was dissolved into 100 ml of acetonitrile, 4-(4-pyridyl) pip ucts, and treated by chromatography with neutral Al-O. erazine (4.9 g, 0.03 mol), diisopropylethylamine (15.5g, 0.12 eluted with dichloromethane/methanol mixture to produce mol) and potassium iodide (5.0 g, 0.03 mol) were respectively 6.8 g compound (I-16) with a yield of 59.1%. ESI-MS added. The mixture was stirred for 10 min at ambient tem M+H: m/z 385.2. perature, and then heated and refluxed to react for 20 hours. The mixture was cooled down to ambient temperature and Example 17 filtered. The filtrate was concentrated to produce oily prod ucts, and treated by chromatography with neutral Al2O, Preparation of 1-(4-(4-(2-quinoxalinyl)piperazine-1- eluted with dichloromethane/methanol mixture to produce 10 yl)butyl)-1H-benzimidazole (I-17) 6.3 g compound (I-13) with a yield of 62.1%. ESI-MS M+H: m/z 336.2. The method described in Example 1 was adopted to pre Example 14 pare 1-(4-chlorobutyl)-1H-benzimidazole. 15 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) Preparation of 1-(4-(4-(2-pyrimidinyl)piperazine-1- was dissolved into 100 ml of acetonitrile, 4-(2-quinoxalinyl) yl)butyl)-1H-benzimidazole (I-14) piperazine (6.4g, 0.03 mol), diisopropylethylamine (15.5 g. 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec The method described in Example 1 was adopted to pre tively added. The mixture was stirred for 10 min at ambient pare 1-(4-chlorobutyl)-1H-benzimidazole. temperature, and then heated and refluxed to react for 20 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) hours. The mixture was cooled down to ambient temperature was dissolved into 100 ml of acetonitrile, 4-(2-pyrimidinyl) and filtered. The filtrate was concentrated to produce oily piperazine (4.9 g, 0.03 mol), diisopropylethylamine (15.5 g. products, and treated by chromatography with neutral Al-O. 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec eluted with dichloromethane/methanol mixture to produce tively added. The mixture was stirred for 10 min at ambient 25 6.9 g compound (I-17) with a yield of 59.6%. ESI-MS temperature, and then heated and refluxed to react for 20 M+H": m/z 387.2. hours. The mixture was cooled down to ambient temperature and filtered. The filtrate was concentrated to produce oily Example 18 products, and treated by chromatography with neutral Al-O. eluted with dichloromethane/methanol mixture to produce 30 Preparation of 1-(4-(4-(3-(6-fluorobenzisoxazolyl)) 6.1 g compound (I-14) with a yield of 60.1%. ESI-MS piperazine-1-yl)butyl)-1H-benzimidazole (I-18) M+H: m/z 337.2. Example 15 The method described in Example 1 was adopted to pre 35 pare 1-(4-chlorobutyl)-1H-benzimidazole. Preparation of 1-(4-(4-(1-cyclohexyl)piperazine-1- 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) yl)butyl)-1H-benzimidazole (I-15) was dissolved into 100 ml of acetonitrile, 6-fluoro-3-(pipera Zine-4-yl)benzisoxazolyl (6.6 g., 0.05 mol), diisopropylethy The method described in Example 1 was adopted to pre lamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 pare 1-(4-chlorobutyl)-1H-benzimidazole. 40 mol) were respectively added. The mixture was stirred and 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) mixed, and then heated and refluxed to react for 15 hours. The was dissolved into 100 ml of acetonitrile, 4-(1-cyclohexyl) mixture was cooled down to ambient temperature and fil piperazine (5.1 g, 0.03 mol), diisopropylethylamine (15.5 g. tered. The filtrate was concentrated to produce oily products, 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec and treated by chromatography with neutral Al-O and puri tively added. The mixture was stirred for 10 min at ambient 45 fied, eluted with dichloromethane to produce 7.7 g compound temperature, and then heated and refluxed to react for 20 (I-18) with a yield of 65.6%. ESI-MS M+H": m/z 394.2. hours. The mixture was cooled down to ambient temperature and filtered. The filtrate was concentrated to produce oily Example 19 products, and treated by chromatography with neutral Al2O, eluted with dichloromethane/methanol mixture to produce 50 Preparation of 1-(4-(4-(3-(6-fluorobenzisothiazolyl)) 6.4 g compound (I-15) with a yield of 62.9%. ESI-MS piperazine-1-yl)butyl)-1H-benzimidazole (I-19) M+H": m/z 341.3. Example 16 The method described in Example 1 was adopted to pre 55 pare 1-(4-chlorobutyl)-1H-benzimidazole. Preparation of 1-(4-(4-(1-naphthyl)piperazine-1-yl) 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) butyl)-1H-benzimidazole (I-16) was dissolved into 100 ml of acetonitrile, 6-fluoro-3-(pipera Zine-4-yl)benzisothiazolyl (7.1 g, 0.05 mol), diisopropyl The method described in Example 1 was adopted to pre ethylamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, pare 1-(4-chlorobutyl)-1H-benzimidazole. 60 0.03 mol) were respectively added. The mixture was stirred 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) and mixed, and then heated and refluxed to react for 15 hours. was dissolved into 100 ml of acetonitrile, 4-(1-naphthyl)pip The mixture was cooled down to ambient temperature and erazine (6.4g, 0.03 mol), diisopropylethylamine (15.5g, 0.12 filtered. The filtrate was concentrated to produce oily prod mol) and potassium iodide (5.0 g, 0.03 mol) were respectively ucts, and treated by chromatography with neutral Al2O and added. The mixture was stirred for 10 min at ambient tem 65 purified, eluted with dichloromethane to produce 7.9 g com perature, and then heated and refluxed to react for 20 hours. pound (I-19) with a yield of 64.6%. ESI-MS M+H": m/z. The mixture was cooled down to ambient temperature and 410.2. US 9,415,047 B2 59 60 Example 20 ucts, and treated by chromatography with neutral Al-O puri fied, eluted with dichloromethane/methanol mixture to pro Preparation of 1-(4-(4-(3-benzimidazoyl)piperazine duce 13.7 g 1-(3-(4-(3-(6-fluorobenzisoxazolyl)) piperazine 1-yl)butyl)-1H-benzimidazole (I-20) 1-yl)propoxyl)-1H-benzimidazole (I-22) with a yield of 69.1%. ESI-MSM--H: m/z 396.2. The method described in Example 1 was adopted to pre pare 1-(4-chlorobutyl)-1H-benzimidazole. Example 23 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, 3-(piperazine-4-yl) Preparation of 1-(4-(4-(3-trifluoromethylphenyl) benzimidazole (6.1 g, 0.05 mol), diisopropylethylamine 10 piperazine-1-yl)propoxyl)-1H-benzimidazole (I-23) (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were respectively added. The mixture was stirred and mixed, The method described in Example 22 was adopted to pre and then heated and refluxed to react for 15 hours. The mix pare 1-(3-chloropropoxyl)benzimidazole. ture was cooled down to ambient temperature and filtered. 1-(3-chloropropoxyl)benzimidazole (0.06 mol) was dis The filtrate was concentrated to produce oily products, and 15 solved into 150 ml of acetonitrile, 4-(3-(3-trifluorometh treated by chromatography with neutral Al-O and purified, ylphenyl) piperazine (0.05 mol), diisopropylethylamine (0.2 eluted with dichloromethane to produce 6.9 g compound mol) and potassium iodide (0.05 mol) were respectively (I-20) with a yield of 61.5%. ESI-MSM--HI": m/z. 375.2. added. The mixture was stirred for 10 min at ambient tem perature, and then heated and refluxed to react for 15 hours. Example 21 The mixture was cooled down to ambient temperature and filtered. The filtrate was concentrated to produce oily prod Preparation of 1-(4-(4-(3-(6-fluorobenzofuranyl)) ucts, and treated by chromatography with neutral Al-O puri piperazine-1-yl)butyl)-1H-benzimidazole (I-21) fied, eluted with dichloromethane/methanol mixture to pro duce 13.7 g 1-(4-(4-(3-trifluoromethylphenyl)piperazine-1- The method described in Example 1 was adopted to pre 25 yl)propoxyl)-1H-benzimidazole (I-23) with a yield of 67.9%. pare 1-(4-chlorobutyl)-1H-benzimidazole. ESI-MSM--H: m/z 405.2. 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, 6-fluoro-3-(pipera Example 24 Zine-4-yl)benzofuranyl (6.6 g., 0.05 mol), diisopropylethy lamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 30 Preparation of 1-(4-(4-(3-chlorphenyl)piperazine-1- mol) were respectively added. The mixture was stirred and yl)propoxyl)-1H-benzimidazole (I-24) mixed, and then heated and refluxed to react for 15 hours. The mixture was cooled down to ambient temperature and fil The method described in Example 22 was adopted to pre tered. The filtrate was concentrated to produce oily products, pare 1-(3-chloropropoxyl)benzimidazole. and treated by chromatography with neutral Al-O and puri 35 1-(3-chloropropoxyl)benzimidazole (0.06 mol) was dis fied, eluted with dichloromethane to produce 7.5 g compound solved into 150 ml of acetonitrile, 4-(3-(3-chlorophenyl) pip (I-21) with a yield of 63.6%. ESI-MS M+H": m/z 393.2. erazine (0.05 mol), diisopropylethylamine (0.2 mol) and potassium iodide (0.05 mol) were respectively added. The Example 22 mixture was stirred for 10 min at ambient temperature, and 40 then heated and refluxed to react for 15 hours. The mixture Preparation of 1-(3-(4-(3-(6-fluorobenzisoxazolyl)) was cooled down to ambient temperature and filtered. The piperazine-1-yl)propoxyl)-1H-benzimidazole (I-22) filtrate was concentrated to produce oily products, and treated by chromatography with neutral Al2O, purified, eluted with Preparation of 1-(3-chloro propoxyl)benzimidazole dichloromethane/methanol mixture to produce 12.2 g 1-(4- 45 (4-(3-fluorophenyl)piperazine-1-yl)propoxyl)-1H-benzimi Substituted 1-hydroxylbenzimidazole (0.01 mol) was dis dazole (I-24) with a yield of 66.1%. ESI-MS M+H": m/z solved in 10 ml of NMP solid paraffin mixture containing 3712. 50% (w/w) hydrogen and oxygen was added in different times, stirred to react for 0.5 h. Meanwhile, 3-bromochloro Example 25 propane (0.015 mol) was dissolved in 5 ml of NMP and added 50 into the above said solution, and stirred to react for 12 h. Preparation of 6-chloro-1-(4-(4-(3-trifluorometh Reaction solution was poured into 50 ml of water, extracted ylphenyl)piperazine-1-yl)butyl)-1H-benzimidazole with ethyl acetate (3x50 mL). Organic phases were mixed and (I-25) washed with 30 ml of water. Anhydrous magnesium sulfate was added to dry organic phase, filtered, with solvent evapo 55 6-chloro-1H-benzimidazole (15.2 g, 0.10 mol) was dis rated. Oily products were analyzed by chromatography with solved into 200 ml of 20% wt. sodium hydroxide, 4-chloro neutral Al-O, or separated and purified by using HPLC to bromobutane (34.3 g, 0.20 mol) and tetrabutyl ammonium prepare 1-(3-chloropropoxyl)benzimidazole, with a yield of bromide (1.0 g) were added, and mixed for 5 min. The mix 75.0%. ture was heated to 60° C., stirred to react for 2 hours. Post 1-(3-chloropropoxyl)benzimidazole (0.06 mol) was dis 60 treatment was performed based on common method one for solved into 150 ml of acetonitrile, 4-(3-(6-fluorobenzisox synthesis. Oily products were separated and purified by chro azolyl)) piperazine (0.05 mol), diisopropylethylamine (0.2 matography with neutral Al-O to produce 15.1 g of 1-(4- mol) and potassium iodide (0.05 mol) were respectively chlorobutyl)-6-chloro-1H-benzimidazole, with a yield of added. The mixture was stirred for 10 min at ambient tem 62.3%. perature, and then heated and refluxed to react for 15 hours. 65 1-(4-chlorobutyl)-6-chloro-1H-benzimidazole (8.71 g, The mixture was cooled down to ambient temperature and 0.036 mol) was dissolved into 100 ml of acetonitrile, filtered. The filtrate was concentrated to produce oily prod 3-trichloromethyl phenylpiperazine (6.91 g, 0.03 mol), diiso US 9,415,047 B2 61 62 propylethylamine (15.5 g., 0.12 mol) and potassium iodide nol mixture to produce 8.8 g compound (I-27) with a yield of (5.0 g, 0.03 mol) were respectively added. The mixture was 63.7%. ESI-MSM--H: m/z 461.2. stirred for 10 minatambient temperature, and then heated and refluxed to react for 15 hours. The mixture was cooled down Example 28 to ambient temperature and filtered. The filtrate was concen 5 trated to produce oily products, and treated by chromatogra Preparation of 2-chloro-1-(5-(4-(3-trifluorometh phy with neutral Al-O, eluted with dichloromethane/metha ylphenyl)piperazine-1-yl)pentyl)-1H-benzimidazole nol mixture to produce 8.6 g compound (I-25) with a yield of (I-28) 65.8%. ESI-MSM--H: m/z 437.2. 10 2-chloro-1H-benzimidazole (15.2 g, 0.10 mol) was dis Example 26 solved into 200 ml of 20% wt. sodium hydroxide, 5-chloro bromopentane (36.8 g., 0.20 mol) and tetrabutyl ammonium Preparation of 6-cyano-1-(4-(4-(3-trifluorometh bromide (1.0 g) were added, and mixed for 5 min. The mix ylphenyl)piperazine-1-yl)butyl)-1H-benzimidazole ture was heated to 60°C., stirred to react for 2 hours. Then the (I-26) 15 reaction Solution was cooled down to ambient temperature, 100 ml of dichloromethane was added for extraction and 6-cyano-1H-benzimidazole (14.3 g, 0.10 mol) was dis liquid separation. To the aqueous phase, 100 of dichlo solved into 200 ml of 20% wt. sodium hydroxide, 4-chloro romethane was added for extraction. Organic phases were bromobutane (34.3 g, 0.20 mol) and tetrabutyl ammonium mixed, washed with 100 ml of saturated saline. Liquid was bromide (1.0 g) were added, and mixed for 5 min. The mix separated, and organic phase was evaporated to dryness to ture was heated to 60° C., stirred to react for 2 hours. Post produce oily product. Oily products were separated and puri treatment was performed based on common method one for fied by chromatography with neutral Al-O to produce 16.0g synthesis. Oily products were separated and purified by chro of 1-(5-chloropentyl)-2-chloro-1H-benzimidazole, with a matography with neutral Al-O to produce 14.7 g of 1-(4- 25 yield of 62.5%. chlorobutyl)-6-cyano-1H-benzimidazole, with a yield of 1-(5-chloropentyl)-2-chloro-1H-benzimidazole (9.22 g, 63.1%. 0.036 mol) was dissolved into 100 ml of acetonitrile, 1-(4-chlorobutyl)-6-cyano-1H-benzimidazole (8.39 g, 3-trichloromethyl phenylpiperazine (6.91 g, 0.03 mol), diiso 0.036 mol) was dissolved into 100 ml of acetonitrile, propylethylamine (15.5 g., 0.12 mol) and potassium iodide 3-trichloromethyl phenylpiperazine (6.91 g, 0.03 mol), diiso 30 (5.0 g, 0.03 mol) were respectively added. The mixture was propylethylamine (15.5 g., 0.12 mol) and potassium iodide stirred for 10 minatambient temperature, and then heated and (5.0 g, 0.03 mol) were respectively added. The mixture was refluxed to react for 15 hours. The mixture was cooled down stirred for 10 minatambient temperature, and then heated and to ambient temperature and filtered. The filtrate was concen refluxed to react for 15 hours. The mixture was cooled down trated to produce oily products, and treated by chromatogra to ambient temperature and filtered. The filtrate was concen 35 phy with neutral Al-O, eluted with dichloromethane/metha trated to produce oily products, and treated by chromatogra nol mixture to produce 8.8 g compound (I-28) with a yield of phy with neutral Al-O, eluted with dichloromethane/metha 65.2%. ESI-MSM--H: m/z 451.2. nol mixture to produce 8.6 g compound (I-26) with a yield of 66.9%. ESI-MSM--H: m/z 428.2. 40 Example 29 Example 27 Preparation of 1-(4-(4-(3-chlorophenyl)piperazine-1- yl)butyl)-1H-benzotriazole (I-29) Preparation of 6-methoxycarbonyl-1-(4-(4-(3-trifluo romethylphenyl)piperazine-1-yl)butyl)-1H-benzimi 45 Benzotriazole (11.9 g, 0.10 mol) is dissolved into 100 ml of dazole (I-27) 30% wt. sodium hydroxide, 4-chlorobromobutane (34.3 g, 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are added, 6-methoxycarbonyl-1H-benzimidazole (17.6 g., 0.10 mol) and mixed for 5 min. The reaction solution is gradually heated was dissolved into 200 ml of 20% wt. sodium hydroxide, to 60° C., stirred for reaction for 2 hours. Then the reaction 4-chlorobromobutane (34.3 g, 0.20 mol) and tetrabutyl 50 solution was cooled down to ambient temperature, 100 ml of ammonium bromide (1.0 g) were added, and mixed for 5 min. dichloromethane was added for extraction and liquid separa The mixture was heated to 60°C., stirred to react for 2 hours. tion. To the aqueous phase, 100 of dichloromethane was Post treatment was performed based on common method one added for extraction. Organic phases were mixed, washed for synthesis. Oily products were separated and purified by with 100 ml of saturated saline. Liquid was separated, and chromatography with neutral Al-O to produce 16.9 g of 55 organic phase was evaporated to dryness to produce oily 1-(4-chlorobutyl)-6-methoxycarbonyl-1H-benzimidazole, product. Oily products were separated and purified by chro with a yield of 63.4%. matography with neutral Al-O, and eluted with dichlo 1-(4-chlorobutyl)-6-methoxycarbonyl-1H-benzimidazole rmethane to produce 17.0 g of 1-(4-chlorobutyl)-1H-benzot (9.58 g., 0.036 mol) was dissolved into 100 ml of acetonitrile, riazole, with a yield of 81.0%. 3-trichloromethyl phenylpiperazine (6.91 g, 0.03 mol), diiso 60 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) propylethylamine (15.5 g., 0.12 mol) and potassium iodide was dissolved into 100 ml of acetonitrile, 3-trichlorophe (5.0 g, 0.03 mol) were respectively added. The mixture was nylpiperazine (5.9 g, 0.03 mol), diisopropylethylamine (15.5 stirred for 10 minatambient temperature, and then heated and g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were refluxed to react for 15 hours. The mixture was cooled down respectively added. The mixture was stirred and mixed, then to ambient temperature and filtered. The filtrate was concen 65 heated and refluxed to react for 15 hours. The mixture was trated to produce oily products, and treated by chromatogra cooled down to ambient temperature and filtered. The filtrate phy with neutral Al-O, eluted with dichloromethane/metha was concentrated to produce oily products, and treated by US 9,415,047 B2 63 64 chromatography with neutral Al-O and purified, eluted with Post treatment was performed based on common method one dichloromethane to produce 7.8 g. compound (I-29) with a for synthesis. The solution was separated and purified by yield of 70.3%. HPLC to produce 8.9 g of 1-(4-chlorobutyl)-6-fluoro-1H Compound (I-29) (5.55 g, 0.015 mol) was dissolved in 50 benzotriazole, with a yield of 39.0%. ml of ethyl acetate. Under cooling conditions of icy water 5 1-(4-chlorobutyl)-6-fluoro-1H-benzotriazole (8.2g, 0.036 bath, 3 mol/L hydrogen chloride/ethyl acetate solution is mol) was dissolved into 100 ml of acetonitrile, 3-trifluoro dripped, and the pH value is adjusted to 2. The mixture is methylphenylpiperazine (6.9 g, 0.03 mol), diisopropylethy stirred for 10 min, filtered and dried to produce 5.4 g solid lamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 compound (II-29) with a yield of 88.0%. ESI-MS M+H": mol) were respectively added. The mixture was stirred and m/Z 370.1. 10 mixed, then heated and refluxed to react for 15 hours. The mixture was cooled down to ambient temperature and fil Example 30 tered. The filtrate was concentrated to produce oily products, Preparation of 1-(4-(4-(3-fluorophenyl)piperazine-1- and treated by chromatography with neutral Al-O and puri 15 fied, eluted with dichloromethane to produce 8.3 g compound yl)butyl)-1H-benzotriazole (I-30) (I-32) with a yield of 65.7%. ESI-MS M+H": m/z 422.2. The method described in Example 29 was adopted to pre pare 1-(4-chlorobutyl)-1H-benzotriazole. Example 33 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, 3-trifluorophe Preparation of 5,6-dimethyl-1-(4-(4-(3-trifluorom nylpiperazine (5.4g, 0.03 mol), diisopropylethylamine (15.5 ethylphenyl)piperazine-1-yl)butyl)-1H-benzotriazole g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were (I-33) respectively added. The mixture was stirred and mixed, then heated and refluxed to react for 15 hours. The mixture was 5,6-dimethyl-1H-benzotriazole (14.7 g., 0.10 mol) is dis cooled down to ambient temperature and filtered. The filtrate 25 solved into 100 ml of 30% wt. sodium hydroxide, 4-chloro was concentrated to produce oily products, and treated by bromobutane (34.3 g, 0.20 mol), tetrabutyl ammonium bro chromatography with neutral Al-O and purified, eluted with mide (0.8 g) are added, and mixed for 5 min. The reaction dichloromethane to produce 7.3 g compound (I-30) with a solution is gradually heated to 60°C., stirred for reaction for yield of 68.9%. ESI-MS M+H: m/z. 354.2. 2 hours. Post treatment was performed based on common 30 method one for synthesis. The solution was separated and Example 31 purified by HPLC to produce 17.4 g of 1-(4-chlorobutyl)-5, 6-dimethyl-1H-benzotriazole, with a yield of 73.2%. Preparation of 1-(4-(4-(3-trifluoromethylphenyl) 1-(4-chlorobutyl)-5,6-dimethyl-1H-benzotriazole (8.56 g. piperazine-1-yl)butyl)-1H-benzotriazole (I-31) and 0.036 mol) was dissolved into 100 ml of acetonitrile, 3-trif 1-(4-(4-(3-trifluoromethylphenyl)piperazine-1-yl) 35 luoro methylphenylpiperazine (6.9 g, 0.03 mol), diisopropy butyl)-1H-benzotriazole (II-31) lethylamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) 0.03 mol) were respectively added. The mixture was stirred was dissolved into 100 ml of acetonitrile, 3-trifluoro meth and mixed, then heated and refluxed to react for 15 hours. The ylphenylpiperazine (6.9 g, 0.03 mol), diisopropylethylamine 40 mixture was cooled down to ambient temperature and fil (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) tered. The filtrate was concentrated to produce oily products, were respectively added. The mixture was stirred and mixed, and treated by chromatography with neutral Al-O and puri then heated and refluxed to react for 15 hours. The mixture fied, eluted with dichloromethane to produce 9.1 g compound was cooled down to ambient temperature and filtered. The (I-33) with a yield of 70.3%. ESI-MS M+H": m/z. 432.2. filtrate was concentrated to produce oily products, and treated 45 by chromatography with neutral Al2O and purified, eluted Example 34 with dichloromethane to produce 7.8 g. compound (I-31) with a yield of 64.5%. Preparation of 3-(4-(4-(1H-benzotriazole-1-yl)butyl) Compound (I-31) (6.05 g, 0.015 mol) was dissolved in 50 piperazine-1-yl)benzisothiazole (I-34) ml of ethyl acetate. Under cooling conditions of icy water 50 bath, 3 mol/L hydrogen chloride/ethyl acetate solution is The method described in Example 29 was adopted to pre dripped, and the pH value is adjusted to 2. The mixture is pare 1-(4-chlorobutyl)-1H-benzotriazole. stirred for 10 min, filtered and dried to produce 5.6 g solid 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) compound (II-31) with a yield of 84.8%. ESI-MS M+H": was dissolved into 100 ml of acetonitrile, 3-(piperazine-1-yl) m/z, 404.2. 55 benzisothiazole (6.58 g., 0.03 mol), diisopropylethylamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) Example 32 were respectively added. The mixture was stirred and mixed, then heated and refluxed to react for 15 hours. The mixture Preparation of 6-fluoro-1-(4-(4-(3-trifluorometh was cooled down to ambient temperature and filtered. The ylphenyl)piperazine-1-yl)butyl)-1H-benzotriazole 60 filtrate was concentrated to produce oily products, and treated (I-32) by chromatography with neutral Al-O and purified, eluted with dichloromethane to produce 8.2 g compound (I-34) with 6-fluoro-1H-benzotriazole (13.7g, 0.10 mol) is dissolved a yield of 69.6%. into 100 ml of 30% wt. sodium hydroxide, 4-chlorobromobu Compound (I-34) (5.89 g, 0.015 mol) was dissolved in 50 tane (34.3g, 0.20 mol), tetrabutyl ammonium bromide (0.8 g) 65 ml of ethyl acetate and 5 ml of ethanol. Under cooling con are added, and mixed for 5 min. The reaction solution is ditions of icy water bath, 3 mol/L hydrogen chloride/ethyl gradually heated to 60° C., stirred for reaction for 2 hours. acetate solution is dripped, and the pH value is adjusted to 2. US 9,415,047 B2 65 66 The mixture is stirred for 10 min, filtered and dried to produce mixed, then heated and refluxed to react for 15 hours. The 5.5g solid compound (II-34) with a yield of 85.5%. ESI-MS mixture was cooled down to ambient temperature and fil M+H: m/z 393.2. tered. The filtrate was concentrated to produce oily products, and treated by chromatography with neutral Al-O and puri Example 35 fied, eluted with dichloromethane to produce 7.9 g compound (I-37) with a yield of 69.3%. ESI-MSM--HI: m/z 380.2. Preparation of 3-(4-(4-(1H-benzotriazole-1-yl)butyl) piperazine-1-yl)benzisoxazolyl (I-35) Example 38 Preparation of 1-(3-(4-(2,3-dichlorophenyl)pipera The method described in Example 29 was adopted to pre 10 Zine-1-yl)butyl)-1H-benzotriazole (I-38) pare 1-(4-chlorobutyl)-1H-benzotriazole. 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) The method described in Example 37 was adopted to pre was dissolved into 100 ml of acetonitrile, 3-(piperazine-1-yl) pare 1-(3-chloropropyl)-1H-benzotriazole. benzisoxazole (6.1 g, 0.03 mol), diisopropylethylamine (15.5 1-(3-chloropropyl)-1H-benzotriazole (7.02 g, 0.036 mol) g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were 15 was dissolved into 100 ml of acetonitrile, 2,3-dicholorophe respectively added. The mixture was stirred and mixed, then nylpiperazine (6.9 g, 0.03 mol), diisopropylethylamine (15.5 heated and refluxed to react for 15 hours. The mixture was g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were cooled down to ambient temperature and filtered. The filtrate respectively added. The mixture was stirred and mixed, then was concentrated to produce oily products, and treated by heated and refluxed to react for 15 hours. The mixture was chromatography with neutral Al-O and purified, eluted with cooled down to ambient temperature and filtered. The filtrate dichloromethane to produce 8.0 g compound (I-35) with a was concentrated to produce oily products, and treated by chromatography with neutral Al-O and purified, eluted with yield of 70.9%. ESI-MS M+H: m/z. 377.2. dichloromethane to produce 8.2 g compound (I-38) with a Example 36 yield of 70.2%. ESI-MS M+H": m/z 389.1. 25 Preparation of 6-fluoro-3-(4-(4-(1H-benzotriazole-1- Example 39 yl)butyl)piperazine-1-yl)benzisoxazole (I-36) Preparation of 1-(3-(4-(3-methylphenyl)piperazine-1-yl) butyl)-1H-benzotriazole (I-39) The method described in Example 29 was adopted to pre The method described in Example 37 was adopted to pre pare 1-(4-chlorobutyl)-1H-benzotriazole. 30 pare 1-(3-chloropropyl)-1H-benzotriazole. 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) 1-(3-chloropropyl)-1H-benzotriazole (7.02 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, 6-fluoro-3-(pipera was dissolved into 100 ml of acetonitrile, 3-methyl phe Zine-1-yl)benzisoxazole (6.1 g, 0.03 mol), diisopropylethy nylpiperazine (5.3.g., 0.03 mol), diisopropylethylamine (15.5 lamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were mol) were respectively added. The mixture was stirred and 35 respectively added. The mixture was stirred and mixed, then mixed, then heated and refluxed to react for 15 hours. The heated and refluxed to react for 15 hours. The mixture was mixture was cooled down to ambient temperature and fil cooled down to ambient temperature and filtered. The filtrate tered. The filtrate was concentrated to produce oily products, was concentrated to produce oily products, and treated by and treated by chromatography with neutral Al-O and puri chromatography with neutral Al-O and purified, eluted with fied, eluted with dichloromethane to produce 8.3 g compound 40 dichloromethane to produce 7.5 g compound (I-39) with a (I-36) with a yield of 70.0%. ESI-MSM--HI": m/z 395.2. yield of 74.6%. ESI-MS M+H: m/z 335.2. Example 37 Example 40 Preparation of 1-(4-(4-(3-methoxyphenyl)piperazine Preparation of 6-fluoro-3-(4-(3-(1H-benzotriazole-1- 45 yl)propyl)piperazine-1-yl)benzisoxazole (I-37) 1-yl)butyl)-1H-benzotriazole (I-40) The method described in Example 29 was adopted to pre Benzotriazole (11.9 g, 0.10 mol) is dissolved into 100 ml of pare 1-(4-chlorobutyl)-1H-benzotriazole. 30% wt. sodium hydroxide, 3-chlorobromopropane (30.2g, 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are added, 50 was dissolved into 100 ml of acetonitrile, 3-methyl phe and mixed for 5 min. The reaction solution is gradually heated nylpiperazine (5.8 g., 0.03 mol), diisopropylethylamine (15.5 to 60° C., stirred for reaction for 2 hours. Then the reaction g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were solution was cooled down to ambient temperature, 100 ml of respectively added. The mixture was stirred and mixed, then dichloromethane was added for extraction and liquid separa heated and refluxed to react for 15 hours. The mixture was tion. To the aqueous phase, 100 of dichloromethane was 55 cooled down to ambient temperature and filtered. The filtrate added for extraction. Organic phases were mixed, washed was concentrated to produce oily products, and treated by with 100 ml of saturated saline. Liquid was separated, and chromatography with neutral Al-O and purified, eluted with organic phase was evaporated to dryness to produce oily dichloromethane to produce 7.6 g compound (I-40) with a product. Oily products were separated and purified by chro yield of 69.4%. ESI-MS M+H: m/z. 365.2. matography with neutral Al-O, and eluted with dichlo 60 rmethane to produce 15.6 g of 1-(3-chloropropyl)-1H-benzo Example 41 triazole, with a yield of 80.0%. 1-(3-chloropropyl)-1H-benzotriazole (7.02 g, 0.036 mol) Preparation of 1-(4-(4-(3-cyanophenyl)piperazine-1- was dissolved into 100 ml of acetonitrile, 6-fluoro-3-(pipera yl)butyl)-1H-benzotriazole (I-41) Zine-1-yl)benzisoxazole (6.6 g., 0.03 mol), diisopropylethy 65 lamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 The method described in Example 29 was adopted to pre mol) were respectively added. The mixture was stirred and pare 1-(4-chlorobutyl)-1H-benzotriazole. US 9,415,047 B2 67 68 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, 3-cyano phenylpip was dissolved into 100 ml of acetonitrile, 4-(4-pyridyl)pip erazine (5.6 g., 0.03 mol), diisopropylethylamine (15.5g, 0.12 erazine (4.9 g, 0.03 mol), diisopropylethylamine (15.5g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were respectively mol) and potassium iodide (5.0g, 0.03 mol) were respectively added. The mixture was stirred and mixed, then heated and refluxed to react for 15 hours. The mixture was cooled down added. The mixture was stirred and mixed, then heated and to ambient temperature and filtered. The filtrate was concen refluxed to react for 20 hours. The mixture was cooled down trated to produce oily products, and treated by chromatogra to ambient temperature and filtered. The filtrate was concen phy with neutral Al-O and purified, eluted with dichlo trated to produce oily products, and treated by chromatogra romethane to produce 7.6 g compound (I-41) with a yield of phy with neutral Al-O and purified, eluted with dichlo 70.5%. ESI-MSM--H": m/z 360.2. 10 romethane to produce 6.6 g compound (I-44) with a yield of 65.3%. ESI-MSM--H: m/z 336.2. Example 42 Example 45 Preparation of 1-(5-(4-(3-trifluoromethylphenyl) piperazine-1-yl)pentyl)-1H-benzotriazole (I-42) 15 Preparation of 1-(4-(4-(2-pyrimidinyl)piperazine-1- yl)butyl)-1H-benzotriazole (I-45) Benzotriazole (11.9 g, 0.10 mol) is dissolved into 100 ml of 30% wt. sodium hydroxide, 5-chlorobromopentane (36.8 g. 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are added, The method described in Example 29 was adopted to pre and mixed for 5 min. The reaction solution is gradually heated pare 1-(4-chlorobutyl)-1H-benzotriazole. to 60° C., stirred for reaction for 2 hours. Then the reaction 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) solution was cooled down to ambient temperature, 100 ml of was dissolved into 100 ml of acetonitrile, 4-(2-pyrimidinyl) dichloromethane was added for extraction and liquid separa piperazine (4.9 g, 0.03 mol), diisopropylethylamine (15.5 g. tion. To the aqueous phase, 100 of dichloromethane was 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec added for extraction. Organic phases were mixed, washed with 100 ml of saturated saline. Liquid was separated, and 25 tively added. The mixture was stirred and mixed, then heated organic phase was evaporated to dryness to produce oily and refluxed to react for 20 hours. The mixture was cooled product. Oily products were separated and purified by chro down to ambient temperature and filtered. The filtrate was matography with neutral Al-O, and eluted with dichlo concentrated to produce oily products, and treated by chro rmethane to produce 15.8g of 1-(5-chloropentyl)-1H-benzo matography with neutral Al-O and purified, eluted with triazole, with a yield of 71.0%. 30 dichloromethane to produce 6.8 g compound (I-45) with a 1-(5-chloropentyl)-1H-benzotriazole (8.0 g, 0.036 mol) yield of 67.1%. ESI-MS M+H: m/z 337.2. was dissolved into 100 ml of acetonitrile, 3-trifluoro meth ylphenylpiperazine (6.9 g, 0.03 mol), diisopropylethylamine Example 46 (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were respectively added. The mixture was stirred and mixed, 35 Preparation of 1-(4-(4-cyclohexyl piperazine-1-yl) then heated and refluxed to react for 15 hours. The mixture butyl)-1H-benzotriazole (I-46) was cooled down to ambient temperature and filtered. The filtrate was concentrated to produce oily products, and treated The method described in Example 29 was adopted to pre by chromatography with neutral Al2O and purified, eluted pare 1-(4-chlorobutyl)-1H-benzotriazole. with dichloromethane to produce 7.7 g compound (I-42) with 40 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) a yield of 61.5%. ESI-MSM--HI": m/z. 417.2. was dissolved into 100 ml of acetonitrile, 4-(1-cyclohexyl) piperazine (5.1 g, 0.03 mol), diisopropylethylamine (15.5 g. Example 43 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec Preparation of 1-(4-(4-(2-furyl)piperazine-1-yl)bu tively added. The mixture was stirred and mixed, then heated tyl)-1H-benzotriazole (I-43) 45 and refluxed to react for 20 hours. The mixture was cooled down to ambient temperature and filtered. The filtrate was The method described in Example 29 was adopted to pre concentrated to produce oily products, and treated by chro pare 1-(4-chlorobutyl)-1H-benzotriazole. matography with neutral Al-O and purified, eluted with 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) dichloromethane to produce 6.5 g compound (I-46) with a was dissolved into 100 ml of acetonitrile, 4-(2-furyl)pipera 50 yield of 63.7%. ESI-MS M+H: m/z 341.2. zine (4.6 g., 0.03 mol), diisopropylethylamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were respectively Example 47 added. The mixture was stirred and mixed, then heated and refluxed to react for 20 hours. The mixture was cooled down Preparation of 1-(4-(4-(1-naphthyl)piperazine-1-yl) to ambient temperature and filtered. The filtrate was concen 55 butyl)-1H-benzotriazole (I-47) trated to produce oily products, and treated by chromatogra phy with neutral Al-O and purified, eluted with dichlo The method described in Example 29 was adopted to pre romethane to produce 7.0 g compound (I-43) with a yield of pare 1-(4-chlorobutyl)-1H-benzotriazole. 71.3%. ESI-MSM--H: m/z 325.2. 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) 60 was dissolved into 100 ml of acetonitrile, 4-(1-naphthyl)pip Example 44 erazine (6.4g, 0.03 mol), diisopropylethylamine (15.5g, 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respectively Preparation of 1-(4-(4-(4-pyridyl)piperazine-1-yl) added. The mixture was stirred and mixed, then heated and butyl)-1H-benzotriazole (I-44) refluxed to react for 20 hours. The mixture was cooled down 65 to ambient temperature and filtered. The filtrate was concen The method described in Example 29 was adopted to pre trated to produce oily products, and treated by chromatogra pare 1-(4-chlorobutyl)-1H-benzotriazole. phy with neutral Al-O and purified, eluted with dichlo US 9,415,047 B2 69 70 romethane to produce 6.9 g compound (I-47) with a yield of 1-(3-chloropropyl)-1H-benzotriazole (7.55g, 0.036 mol) 60.1%. ESI-MSM--H: m/z 385.2. was dissolved into 100 ml of acetonitrile, 6-fluoro-3-(pipera Zine-4-yl)benzofuran (6.6 g., 0.05 mol), diisopropylethy Example 48 lamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were respectively added. The mixture was stirred and Preparation of 1-(4-(4-(2-quinoxalinyl)piperazine-1- mixed, then heated and refluxed to react for 20 hours. The yl)butyl)-1H-benzotriazole (I-48) mixture was cooled down to ambient temperature and fil tered. The filtrate was concentrated to produce oily products, The method described in Example 29 was adopted to pre and treated by chromatography with neutral Al-O and puri pare 1-(4-chlorobutyl)-1H-benzotriazole. 10 fied, eluted with dichloromethane to produce 7.9 g compound 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) (I-51) with a yield of 69.1%. ESI-MSM--HI": m/z. 379.2. was dissolved into 100 ml of acetonitrile, 4-(2-quinoxalinyl) piperazine (6.4g, 0.03 mol), diisopropylethylamine (15.5 g. Example 52 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec tively added. The mixture was stirred and mixed, then heated 15 Preparation of 1-(4-(4-(3-(6-fluorobenzisoxazolyl)) and refluxed to react for 20 hours. The mixture was cooled piperazine-1-yl)propoxyl)-1H-benzotriazole (I-52) down to ambient temperature and filtered. The filtrate was concentrated to produce oily products, and treated by chro Preparation of 1-(3-chloro propoxyl)benzotriazole matography with neutral Al-O and purified, eluted with dichloromethane to produce 7.3 g compound (I-48) with a Substituted 1-hydroxyl benzotriazole (0.01 mol) was dis yield of 62.7%. ESI-MS M+H": m/z 387.2. solved in 10 ml of NMP solid paraffin mixture containing 50% (w/w) hydrogen and oxygen was added in different Example 49 times, stirred to react for 0.5 h. Meanwhile, 3-bromochloro propane (0.015 mol) was dissolved in 5 ml of NMP and added Preparation of 1-(4-(4-(3-(6-fluorobenzisothiazolyl)) 25 into the above said solution, and stirred to react for 12 h. piperazine-1-yl)butyl)-1H-benzotriazole (I-49) Reaction solution was poured into 50 ml of water, extracted with ethyl acetate (3x50 mL). Organic phases were mixed and The method described in Example 29 was adopted to pre washed with 30 ml of water. Anhydrous magnesium sulfate pare 1-(4-chlorobutyl)-1H-benzotriazole. was added to dry organic phase, filtered, with solvent evapo 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) 30 rated. Oily products were analyzed by chromatography with was dissolved into 100 ml of acetonitrile, 6-fluoro-3-(pipera neutral Al-O, or separated and purified by using HPLC to zine-4-yl)benzisothiazole (6.6 g., 0.05 mol), diisopropylethy prepare 1-(3-chloropropoxyl)benzotriazole, with a yield of lamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 75.0%. mol) were respectively added. The mixture was stirred and 1-(3-chloropropoxyl)benzotriazole (0.06 mol) was dis mixed, then heated and refluxed to react for 20 hours. The 35 solved into 150 ml of acetonitrile, 4-(3-(6-fluorobenzisox mixture was cooled down to ambient temperature and fil azolyl)) piperazine (0.05 mol), diisopropylethylamine (0.2 tered. The filtrate was concentrated to produce oily products, mol) and potassium iodide (0.05 mol) were respectively and treated by chromatography with neutral Al-O and puri added. The mixture was stirred for 10 min at ambient tem fied, eluted with dichloromethane to produce 8.2 g compound perature, and then heated and refluxed to react for 15 hours. (I-49) with a yield of 66.5%. ESI-MS M+H": m/z 410.2. 40 The mixture was cooled down to ambient temperature and filtered. The filtrate was concentrated to produce oily prod Example 50 ucts, and treated by chromatography with neutral Al-O puri fied, eluted with dichloromethane/methanol mixture to pro Preparation of 1-(4-(4-(3-benzimidazoyl)piperazine duce 13.4 g 1-(4-(4-(3-(6-fluorobenzisoxazolyl)) piperazine 1-yl)butyl)-1H-benzotriazole (I-50) 45 1-yl)propoxyl)-1H-benzotriazole (I-52) with a yield of 67.6%. ESI-MSM--H: m/z 396.2. The method described in Example 29 was adopted to pre pare 1-(4-chlorobutyl)-1H-benzotriazole. Example 53 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, 3-(piperazine-4-yl) 50 Preparation of 6-fluoro-1-(4-(4-(3-(6-fluoro-ben benzopyrazole (6.1 g, 0.05 mol), diisopropylethylamine Zisothiazolyl)piperazine-1-yl) propoxyl)-1H-benzot (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) riazole (I-53) were respectively added. The mixture was stirred and mixed, then heated and refluxed to react for 20 hours. The mixture Preparation of 6-fluoro-1-(2-chloro was cooled down to ambient temperature and filtered. The 55 propoxyl)benzotriazole filtrate was concentrated to produce oily products, and treated by chromatography with neutral Al2O and purified, eluted Substituted 6-fluoro-1-hydroxyl benzotriazole (0.01 mol) with dichloromethane to produce 7.7 g compound (I-50) with was dissolved in 10 ml of NMP solid paraffin mixture (0.01 a yield of 68.2%. ESI-MSM--H: m/z. 375.2. mol) containing 50% (w/w) hydrogen and oxygen was added 60 in different times, stirred to react for 0.5h. Meanwhile, 3-bro Example 51 mochloropropane (0.015 mol) was dissolved in 5 ml of NMP and added into the above said solution, and stirred to react for Preparation of 1-(3-(4-(3-(6-fluorobenzofuranyl)) 12 h. Reaction solution was poured into 50 ml of water, piperazine-1-yl)propyl)-1H-benzotriazole (I-51) extracted with ethyl acetate (3x50 mL). Organic phases were 65 mixed and washed with 30 ml of water. Anhydrous magne The method described in Example 37 was adopted to pre sium sulfate was added to dry organic phase, filtered, with pare 1-(3-chloropropyl)-1H-benzotriazole. solvent evaporated. Oily products were analyzed by chroma US 9,415,047 B2 71 72 tography with neutral Al-O, or separated and purified by produce oily product. Oily products were separated and puri using HPLC to prepare 6-fluoro-1-(3-chloropropoxyl)benzo fied by chromatography with neutral Al-O, and eluted with triazole, with a yield of 75.0%. dichlormethane to produce 17.8g of 6-cyano-1-(4-chlorobu 6-fluoro-1-(3-chloropropoxyl)benzotriazole (0.06 mol) tyl)-1H-benzotriazole, with a yield of 76.0%. was dissolved into 150 ml of acetonitrile, 4-(3-(6-fluoro 6-cyano-1-(4-chlorobutyl)-1H-benzotriazole (8.42 g, benzisothiazolyl)) piperazine (0.05 mol), diisopropylethy 0.036 mol) was dissolved into 100 ml of acetonitrile, 3-trif lamine (0.2 mol) and potassium iodide (0.05 mol) were luoro methylphenylpiperazine (6.9 g, 0.03 mol), diisopropy respectively added. The mixture was stirred for 10 min at lethylamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, ambient temperature, and then heated and refluxed to react for 0.03 mol) were respectively added. The mixture was stirred 15 hours. The mixture was cooled down to ambient tempera 10 and mixed, then heated and refluxed to react for 15 hours. The ture and filtered. The filtrate was concentrated to produce oily mixture was cooled down to ambient temperature and fil products, and treated by chromatography with neutral Al-O. tered. The filtrate was concentrated to produce oily products, purified, eluted with dichloromethane/methanol mixture to and treated by chromatography with neutral Al-O and puri produce 14.1 g 6-fluoro-1-(4-(4-(3-(6-fluoro-benzisothiaz fied, eluted with dichloromethane to produce 8.5 g compound olyl)piperazine-1-yl)propoxyl)-1H-benzotriazole (I-53) 15 (I-55) with a yield of 66.4%. ESI-MS M+H": m/z 428.2. with a yield of 65.6%. ESI-MSM-i-HI": m/z 430.1. Example 56 Example 54 Preparation of 6-methoxycarbonyl-1-(4-(4-(3-trifluo Preparation of 6-chloro-1-(4-(4-(3-trifluorometh romethylphenyl)piperazine-1-yl)butyl)-1H-benzot ylphenyl)piperazine-1-yl)butyl)-1H-benzotriazole riazole (I-56) (I-54) 6-methoxycarbonyl-benzotriazole (17.7 g., 0.10 mol) is 6-chloro-benzotriazole (15.3 g 0.10 mol) is dissolved into dissolved into 100 ml of 30% wt. sodium hydroxide, 4-chlo 100 ml of 30% wt. sodium hydroxide, 4-chlorobromobutane 25 robromobutane (34.3 g, 0.20 mol), tetrabutyl ammonium bro (34.3 g, 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are mide (0.8 g) are added, and mixed for 5 min. The reaction added, and mixed for 5 min. The reaction solution is gradually solution is gradually heated to 60°C., stirred for reaction for heated to 60° C., stirred for reaction for 2 hours. Then the 2 hours. Then the reaction solution was cooled downto ambi reaction solution was cooled down to ambient temperature, ent temperature, 100 ml of dichloromethane was added for 100 ml of dichloromethane was added for extraction and 30 extraction and liquid separation. To the aqueous phase, 100 of liquid separation. To the aqueous phase, 100 of dichlo dichloromethane was added for extraction. Organic phases romethane was added for extraction. Organic phases were were mixed, washed with 100 ml of saturated saline. Liquid mixed, washed with 100 ml of saturated saline. Liquid was was separated, and organic phase was evaporated to dryness separated, and organic phase was evaporated to dryness to to produce oily product. Oily products were separated and produce oily product. Oily products were separated and puri 35 purified by chromatography with neutral Al-O, and eluted fied by chromatography with neutral Al-O, and eluted with with dichlormethane to produce 19.5g of 6-methoxycarbo dichlormethane to produce 19.2 g of 6-chloro-1-(4-chlorobu nyl-1-(4-chlorobutyl)-1H-benzotriazole, with a yield of tyl)-1H-benzotriazole, with a yield of 79.0%. 73.0%. 6-chloro-1-(4-chlorobutyl)-1H-benzotriazole (8.75 g, 6-methoxycarbonyl-1-(4-chlorobutyl)-1H-benzotriazole 0.036 mol) was dissolved into 100 ml of acetonitrile, 3-trif 40 (9.61 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, luoro methylphenylpiperazine (6.9 g, 0.03 mol), diisopropy 3-trifluoro methylphenylpiperazine (6.9 g, 0.03 mol), diiso lethylamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, propylethylamine (15.5 g., 0.12 mol) and potassium iodide 0.03 mol) were respectively added. The mixture was stirred (5.0 g, 0.03 mol) were respectively added. The mixture was and mixed, then heated and refluxed to react for 15 hours. The stirred and mixed, then heated and refluxed to react for 15 mixture was cooled down to ambient temperature and fil 45 hours. The mixture was cooled down to ambient temperature tered. The filtrate was concentrated to produce oily products, and filtered. The filtrate was concentrated to produce oily and treated by chromatography with neutral Al-O and puri products, and treated by chromatography with neutral Al-O fied, eluted with dichloromethane to produce 8.5 g compound and purified, eluted with dichloromethane to produce 8.8 g. (I-54) with a yield of 64.7%. ESI-MS M+H": m/z. 437.2. compound (I-56) with a yield of 63.4%. ESI-MS M+H": 50 m/z. 461.2. Example 55 Example 57 Preparation of 6-cyano-1-(4-(4-(3-trifluorometh ylphenyl)piperazine-1-yl)butyl)-1H-benzotriazole Preparation of 1-(4-(4-(3-trifluoromethylphenyl) (I-55) 55 piperazine-1-yl)butyl)-1H-indole (I-57) 6-cyano-benzotriazole (14.4g, 0.10 mol) is dissolved into 1H-indole (11.7g, 0.10 mol) was dissolved into 200 ml of 100 ml of 30% wt. sodium hydroxide, 4-chlorobromobutane 20% wt. sodium hydroxide, 4-chlorobromobutane (34.3 g, (34.3 g, 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are 0.20 mol) and tetrabutyl ammonium bromide (1.0 g) were added, and mixed for 5 min. The reaction solution is gradually 60 added, and mixed for 5 min. The mixture was heated to 60°C., heated to 60° C., stirred for reaction for 2 hours. Then the stirred to react for 2 hours. Then the reaction solution was reaction solution was cooled down to ambient temperature, cooled down to ambient temperature, 100 ml of dichlo 100 ml of dichloromethane was added for extraction and romethane was added for extraction and liquid separation. To liquid separation. To the aqueous phase, 100 of dichlo the aqueous phase, 100 of dichloromethane was added for romethane was added for extraction. Organic phases were 65 extraction. Organic phases were mixed, washed with 100 ml mixed, washed with 100 ml of saturated saline. Liquid was of Saturated Saline. Liquid was separated, and organic phase separated, and organic phase was evaporated to dryness to was evaporated to dryness to produce oily product. Oily prod US 9,415,047 B2 73 74 ucts were separated and purified by chromatography with 1-(3-chloropropyl)-1H-benzopyrazole (6.98 g., 0.036 mol) neutral Al-O to produce 12.5 g of 1-(4-chlorobutyl)-1H was dissolved into 100 ml of acetonitrile, 3-trifluoromethyl indole, with a yield of 60.1%. phenyl piperazine (6.9 g, 0.03 mol), diisopropylethylamine 1-(4-chlorobutyl)-1H-indole (7.45 g, 0.036 mol) was dis (15.5 g., 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) solved into 100 ml of acetonitrile, 3-trichloromethyl phe were respectively added. The mixture was stirred for 10 min nylpiperazine (6.9 g, 0.03 mol), diisopropylethylamine (15.5 at ambient temperature, and then heated and refluxed to react g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were for 10-15 hours. The mixture was cooled down to ambient respectively added. The mixture was stirred for 10 min at temperature and filtered. The filtrate was concentrated to ambient temperature, and then heated and refluxed to react for produce oily products, and treated by chromatography with 15 hours. The mixture was cooled down to ambient tempera 10 neutral Al-O, eluted with dichloromethane/methanol mix ture and filtered. The filtrate was concentrated to produce oily ture to produce 7.6 g compound (I-59) with a yield of 63.1%. products, and treated by chromatography with neutral Al-O. ESI-MSM--H": m/z 389.2. eluted with dichloromethane/methanol mixture to produce 7.3 g compound (I-57) with a yield of 60.6%. ESI-MS Example 60 M+H": m/z 402.2. 15 Preparation of 6-cyano-1-(3-(4-(2,3-fluorophenyl) Example 58 piperazine-1-yl)propyl)-1H-benzopyrazole (I-60) Preparation of 6-cyano-1-(4-(4-(3-chlorophenyl) 6-cyano-1H-benzopyrazole (14.3 g, 0.10 mol) was dis piperazine-1-yl)butyl)-1H-indole (I-58) solved into 200 ml of 20% wt. sodium hydroxide, 3-chloro bromopropane (31.2g, 0.20 mol) and tetrabutyl ammonium 6-cyano-1H-indole (14.2 g, 0.10 mol) was dissolved into bromide (1.0 g) were added, and mixed for 5 min. The mix 200 ml of 20% wt. sodium hydroxide, 4-chlorobromobutane ture was heated to 60°C., stirred to react for 2 hours. Then the (34.3 g, 0.20 mol) and tetrabutyl ammonium bromide (1.0 g) reaction Solution was cooled down to ambient temperature, were added, and mixed for 5 min. The mixture was heated to 25 100 ml of dichloromethane was added for extraction and 60° C., stirred to react for 2 hours. Then the reaction solution liquid separation. To the aqueous phase, 100 of dichlo was cooled down to ambient temperature, 100 ml of dichlo romethane was added for extraction. Organic phases were romethane was added for extraction and liquid separation. To mixed, washed with 100 ml of saturated saline. Liquid was the aqueous phase, 100 of dichloromethane was added for separated, and organic phase was evaporated to dryness to extraction. Organic phases were mixed, washed with 100 ml 30 produce oily product. Oily products were separated and puri of Saturated Saline. Liquid was separated, and organic phase fied by chromatography with neutral Al-O to produce 13.9 g, was evaporated to dryness to produce oily product. Oily prod of 6-cyano-1-(3-chloropropyl)-1H-benzopyrazole, with a ucts were separated and purified by chromatography with yield of 63.8%. neutral Al-O to produce 13.9 g of 6-cyano-1-(4-chlorobu 6-cyano-1-(3-chloropropyl)-1H-benzimidazole (7.88 g. tyl)-1H-indole, with a yield of 60.3%. 35 0.036 mol) was dissolved into 100 ml of acetonitrile, 2.3- 6-cyano-1-(4-chlorobutyl)-1H-indole (8.35 g, 0.036 mol) dicholoro phenylpiperazine (6.9 g, 0.03 mol), diisopropyl was dissolved into 100 ml of acetonitrile, 3-trichlorophe ethylamine (15.5 g., 0.12 mol) and potassium iodide (5.0 g, nylpiperazine (6.9 g, 0.03 mol), diisopropylethylamine (15.5 0.03 mol) were respectively added. The mixture was stirred g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were for 10 min at ambient temperature, and then heated and respectively added. The mixture was stirred for 10 min at 40 refluxed to react for 15 hours. The mixture was cooled down ambient temperature, and then heated and refluxed to react for to ambient temperature and filtered. The filtrate was concen 15 hours. The mixture was cooled down to ambient tempera trated to produce oily products, and treated by chromatogra ture and filtered. The filtrate was concentrated to produce oily phy with neutral Al-O, eluted with dichloromethane/metha products, and treated by chromatography with neutral Al2O, nol mixture to produce 7.6 g compound (I-60) with a yield of eluted with dichloromethane/methanol mixture to produce 45 61.1%. ESI-MSM--H: m/z 414.1. 6.9 g compound (I-58) with a yield of 62.3%. ESI-MS M+H: m/z 393.2. Example 61 Example 59 The said compounds (I-61 to I-65) in this invention were 50 prepared according to the method described in applying Preparation of 1-(3-(4-(3-trifluoromethylphenyl) patent US20100329978A1. piperazine-1-yl)propyl)-1H-benzopyrazole (I-59) Example 62 1H-benzopyrazole (11.8 g., 0.10 mol) was dissolved into 200 ml of 20% wt. sodium hydroxide, 3-chlorobromopropane 55 The said compounds (I-66 to I-83) in this invention were (31.2g, 0.20 mol) and tetrabutyl ammonium bromide (1.0 g) prepared according to the method described in China patent were added, and mixed for 5 min. The mixture was heated to 20061OO97269.1. 60° C., stirred to react for 2 hours. Then the reaction solution was cooled down to ambient temperature, 100 ml of dichlo Example 63 romethane was added for extraction and liquid separation. To 60 the aqueous phase, 100 of dichloromethane was added for Preparation of N-(3-(1H-benzotriazole-1-yl)propyl)- extraction. Organic phases were mixed, washed with 100 ml 4-(3-benzisoxazolyl)piperidine (I-84) of Saturated Saline. Liquid was separated, and organic phase was evaporated to dryness to produce oily product. Oily prod Benzotriazole (11.9 g, 0.10 mol) is dissolved into 100 ml of ucts were separated and purified by chromatography with 65 30% wt. sodium hydroxide, 3-chlorobromopropane (31.4g, neutral Al-O to produce 11.5 g of 1-(3-chloropropyl)-1H 0.10 mol), tetrabutyl ammonium bromide (0.8 g) are added, benzopyrazole, with a yield of 59.3%. and mixed for 5 min. The reaction solution is gradually heated US 9,415,047 B2 75 76 to 60° C., stirred for reaction for 2 hours. Then the reaction peridine-4-yl)benzisoxazole (11.6 g., 0.05 mol), diisopropyl solution was cooled down to ambient temperature, 100 ml of ethylamine (25.8 g., 0.02 mol) and potassium iodide (8.3 g, dichloromethane was added for extraction and liquid separa 0.05 mol) were respectively added. The mixture was stirred tion. To the aqueous phase, 100 of dichloromethane was and mixed for 10 minatambient temperature, then heated and added for extraction. Organic phases were mixed, washed refluxed to react for 15 hours. After treatment according to with 100 ml of saturated saline. Liquid was separated, and common method three for synthesis produced 13.3 g com organic phase was evaporated to dryness to produce oily pound (I-87), with a yield of 67.7%. ESI-MS M+H": m/z. product. Oily products were separated and purified by chro 392.2. matography with neutral Al-O, and eluted and separated with dichlormethane to produce 16.0 g of 1-(3-chloropropyl)- Example 67 1H-benzotriazole, with a yield of 82%. 10 1-(3-chloropropyl)-1H-benzotriazole (11.7 g., 0.06 mol) Preparation of N-(3-(6-fluoro-1H-benzotriazole-1-yl) was dissolved into 150 ml of acetonitrile, 3-(piperidine-4-yl) propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine benzisoxazole (10.1 g, 0.05 mol), diisopropylethylamine (I-88) (25.8 g., 0.02 mol) and potassium iodide (8.3 g, 0.05 mol) were respectively added. The mixture was stirred and mixed, 15 Preparation of 1-(3-chloro then heated and refluxed to react for 15 hours. The mixture propyl)-6-fluoro-1H-benzotriazole was cooled down to ambient temperature and filtered. The filtrate was concentrated to produce oily products, and treated 6-fluoro-1H-benzotriazole (13.7 g., 0.10 mol) is dissolved by chromatography with neutral Al2O and purified, eluted into 100 ml of 30% wt. sodium hydroxide, 3-chlorobro with dichloromethane to produce 12.6 g compound (I-84) mopropane (31.4g, 0.10 mol), tetrabutyl ammonium bromide with a yield of 69.7%. ESI-MSM-i-HI": m/z. 362.2. (0.8 g) are added, and mixed for 5 min. The reaction solution is gradually heated to 60°C., stirred for reaction for 2 hours. Example 64 Post treatment was performed based on common method one for synthesis. The solution was separated and purified by Preparation of N-(3-(1H-benzotriazole-1-yl)propyl) 25 HPLC to produce 6.9 g of 1-(3-chloropropyl)-6-fluoro-1H 4-(3-(6-fluorobenzisoxazolyl)piperidine (I-85) benzotriazole, with a yield of 32.3%. The method described in Example 63 was adopted to pre Preparation of N-(3-(6-fluoro-1H-benzotriazole-1-yl) pare 1-(3-chloropropyl)-1H-benzotriazole. propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine 1-(3-chloropropyl)-1H-benzotriazole (11.7 g., 0.06 mol) 30 (I-88) was dissolved into 150 ml of acetonitrile, 6-fluoro-3-(piperi dine-4-yl)benzisoxazole (11.0 g, 0.05 mol), diisopropylethy 1-(3-chloropropyl)-6-fluoro-1H-benzotriazole (6.41 g, lamine (25.8 g., 0.02 mol) and potassium iodide (8.3 g 0.05 0.03 mol) was dissolved into 150 ml of acetonitrile, 6-fluoro mol) were respectively added. The mixture was stirred and 3-(piperidine-4-yl)benzisoxazole (5.5 g., 0.025 mol), diiso mixed for 10 min at ambient temperature, then heated and 35 propylethylamine (12.9 g, 0.1 mol) and potassium iodide refluxed to react for 15 hours. The mixture was cooled down (4.15g, 0.025 mol) were respectively added. The mixture was to ambient temperature and filtered. The filtrate was concen stirred and mixed for 10 min at ambient temperature, then trated to produce oily products, and treated by chromatogra heated and refluxed to react for 16 hours. After treatment phy with neutral Al-O, purified, eluted with dichlo according to common method three for synthesis produced romethane/methanol mixture to produce 13.47 g N-(3-(1H 40 8.3 g compound (I-88), with a yield of 69.6%. ESI-MS benzotriazole-1-yl)propyl)4-(3-(6-fluoro benzisoxazolyl)) M+H: m/z 398.2. piperidine (I-85) with a yield of 71.0%. ESI-MSM--HI": m/z. 380.2. Example 68

Example 65 45 Preparation of N-(3-(6-chloro-1H-benzotriazole-1- yl)propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine Preparation of N-(3-(1H-benzotriazole-1-yl)propyl)- (I-89) 4-(3-(6-methylbenzisoxazolyl))piperidine (I-86) Preparation of 1-(3-chloro 1-(3-chloropropyl)-1H-benzotriazole (11.7 g., 0.06 mol) 50 propyl)-6-chloro-1H-benzotriazole was dissolved into 150 ml of acetonitrile, 6-methyl-3-(piperi dine-4-yl)benzisoxazole (10.8 g., 0.05 mol), diisopropylethy 6-chloro-1H-benzotriazole (15.4g, 0.10 mol) is dissolved lamine (25.8 g., 0.02 mol) and potassium iodide (8.3 g 0.05 into 100 ml of 30% wt. sodium hydroxide, 3-chlorobro mol) were respectively added. The mixture was stirred and mopropane (31.4g, 0.10 mol), tetrabutyl ammonium bromide mixed for 10 min at ambient temperature, then heated and 55 (0.8 g) are added, and mixed for 5 min. The reaction solution refluxed to react for 15 hours. After treatment according to is gradually heated to 60°C., stirred for reaction for 2 hours. common method three for synthesis produced 12.4 g com Post treatment was performed based on common method two pound (I-86), with a yield of 66.1%. ESI-MS M+H": m/z. for synthesis. The solution was separated and purified by 376.2. HPLC to produce 7.3 g of 1-(3-chloropropyl)-6-chloro-1H 60 benzotriazole, with a yield of 31.7%. Example 66 Preparation of N-(3-(6-chloro-1H-benzotriazole-1- Preparation of N-(3-(1H-benzotriazole-1-yl)propyl)- yl)propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine 4-(3-(6-methoxyl benzisoxazolyl)piperidine (I-87) (I-89) 65 1-(3-chloropropyl)-1H-benzotriazole (11.7 g., 0.06 mol) 1-(3-chloropropyl)-6-chloro-1H-benzotriazole (6.90 g, was dissolved into 150 ml of acetonitrile, 6-methoxyl-3-(pi 0.03 mol) was dissolved into 150 ml of acetonitrile, 6-fluoro US 9,415,047 B2 77 78 3-(piperidine-4-yl)benzisoxazole (5.5 g., 0.025 mol), diiso 3-(piperidine-4-yl)benzisoxazole (5.5 g., 0.025 mol), diiso propylethylamine (12.9 g, 0.1 mol) and potassium iodide propylethylamine (12.9 g, 0.1 mol) and potassium iodide (4.15g, 0.025 mol) were respectively added. The mixture was (4.15g, 0.025 mol) were respectively added. The mixture was stirred and mixed for 10 min at ambient temperature, then heated and refluxed to react for 16 hours. After treatment stirred and mixed for 10 min at ambient temperature, then according to common method three for synthesis produced heated and refluxed to react for 16 hours. After treatment 8.1 g compound (I-89), with a yield of 65.2%. ESI-MS according to common method three for synthesis produced M+H": m/z. 414.1. 8.6 g compound (I-91), with a yield of 70.0%. ESI-MS M+H": m/z 410.2. Example 69 10 Example 71 Preparation of N-(3-(6-methyl-1H-benzotriazole-1- yl)propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine Preparation of N-(3-(6-formoxyl-1H-benzotriazole (I-90) 1-yl)propyl)-4-(3-(6-fluorobenzisoxazolyl))piperi dine (I-92) Preparation of 1-(3-chloro 15 propyl)-6-methyl-1H-benzotriazole Preparation of 1-(3-chloro propyl)-6-formoxyl-1H-benzotriazole 6-methyl-1H-benzotriazole (13.3 g, 0.10 mol) is dissolved into 100 ml of 30% wt. sodium hydroxide, 3-chlorobro 6-formoxyl-1H-benzotriazole (16.2 g, 0.10 mol) is dis mopropane (31.4g, 0.10 mol), tetrabutyl ammonium bromide solved into 100 ml of 30% wt. sodium hydroxide, 3-chloro (0.8 g) are added, and mixed for 5 min. The reaction solution bromopropane (31.4g, 0.10 mol), tetrabutyl ammonium bro is gradually heated to 60°C., stirred for reaction for 2 hours. mide (0.8 g) are added, and mixed for 5 min. The reaction Post treatment was performed based on common method two solution is gradually heated to 60°C., stirred for reaction for for synthesis. The solution was separated and purified by 2 hours. Post treatment was performed based on common HPLC to produce 7.2 g of 1-(3-chloropropyl)-6-methyl-1H 25 method two for synthesis. The Solution was separated and benzotriazole, with a yield of 34.3%. purified by HPLC to produce 7.9 g of 1-(3-chloropropyl)-6- formoxyl-1H-benzotriazole, with a yield of 33.2%. Preparation of N-(3-(6-methyl-1H-benzotriazole-1- yl)propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine Preparation of N-(3-(6-formoxyl-1H-benzotriazole (I-90) 30 1-yl)propyl)-4-(3-(6-fluorobenzisoxazolyl))piperi dine (I-92) 1-(3-chloropropyl)-6-methyl-1H-benzotriazole (6.29 g, 0.03 mol) was dissolved into 150 ml of acetonitrile, 6-fluoro 1-(3-chloropropyl)-6-formoxyl-1H-benzotriazole (7.13 g, 3-(piperidine-4-yl)benzisoxazole (5.5 g., 0.025 mol), diiso 0.03 mol) was dissolved into 150 ml of acetonitrile, -6-fluoro propylethylamine (12.9 g, 0.1 mol) and potassium iodide 35 3-(piperidine-4-yl)benzisoxazole (5.5 g., 0.025 mol), diiso (4.15g, 0.025 mol) were respectively added. The mixture was propylethylamine (12.9 g, 0.1 mol) and potassium iodide stirred and mixed for 10 min at ambient temperature, then (4.15g, 0.025 mol) were respectively added. The mixture was heated and refluxed to react for 16 hours. After treatment stirred and mixed for 10 min at ambient temperature, then according to common method three for synthesis produced heated and refluxed to react for 15 hours. After treatment 8.5 g compound (I-90), with a yield of 71.9%. ESI-MS 40 according to common method three for synthesis produced M+H": m/z 394.2. 7.5 g compound (I-92), with a yield of 73.6%. ESI-MS M+H: m/z 408.2. Example 70 Example 72 Preparation of N-(3-(6-methoxyl-1H-benzotriazole 45 1-yl)propyl)-4-(3-(6-fluorobenzisoxazolyl))piperi Preparation of N-(3-(1H-benzotriazole-1-yl) propyl)- dine (I-91) 4-(3-(6-fluorobenzisoxazolyl)) piperidine hydro chlorate (II-85) Preparation of 1-(3-chloro propyl)-6-methoxyl-1H-benzotriazole 50 Compound (I-85) (11.38g, 0.03 mol) was dissolved in 100 ml of ethyl acetate and 10 ml of anhydrous ethanol. Under 6-methoxyl-1H-benzotriazole (14.9 g, 0.10 mol) is dis cooling conditions of icy water bath, 3 mol/L hydrogen chlo solved into 100 ml of 30% wt. sodium hydroxide, 3-chloro ride/ethyl acetate solution is dripped, and the pH value is bromopropane (31.4g, 0.10 mol), tetrabutyl ammonium bro adjusted to 2. The mixture is stirred for 10 min, filtered and mide (0.8 g) are added, and mixed for 5 min. The reaction 55 dried to produce 11.4 g solid compound (II-85) with a yield of solution is gradually heated to 60°C., stirred for reaction for 91.2%. 2 hours. Post treatment was performed based on common method two for synthesis. The Solution was separated and Example 73 purified by HPLC to produce 7.7 g of 1-(3-chloropropyl)-6- methoxyl-1H-benzotriazole, with a yield of 34.1%. 60 Preparation of N-(3-(6-methoxyl benzotriazolyl) propyl)-4-(3-benzisoxazolyl)piperidine (I-93) Preparation of N-(3-(6-methoxyl-1H-benzotriazole 1-yl)propyl)-4-(3-(6-fluorobenzisoxazolyl))piperi Preparation of N-(3-chloro dine (I-91) propyl)-6-methoxyl-benzotriazole 65 1-(3-chloropropyl)-6-methoxyl-1H-benzotriazole (6.77g, 6-methoxyl-1H-benzotriazole (14.9 g, 0.10 mol) is dis 0.03 mol) was dissolved into 150 ml of acetonitrile, -6-fluoro solved into 100 ml of 30% wt. sodium hydroxide, 3-chloro US 9,415,047 B2 79 80 bromopropane (31.4g, 0.10 mol), tetrabutyl ammonium bro was cooled down to ambient temperature and filtered. The mide (0.8 g) are added, and mixed for 5 min. The reaction filtrate was concentrated to produce oily products, and treated solution is gradually heated to 60°C., stirred for reaction for by chromatography with neutral Al2O, purified, eluted with 2 hours. Post treatment was performed based on common dichloromethane/methanol mixture to produce 13.96 g N-(4- method two for synthesis. The Solution was separated and (1-benzotriazolyl) butyl)-4-(3-(6-benzisoxazolyl)) piperi purified by HPLC to produce 7.7 g of N-(3-chloropropyl)-6- dine (I-95) with a yield of 71.0%. ESI-MS M+H": m/z methoxyl-benzotriazole, with a yield of 34.1%. 3932. N-(3-chloropropyl)-6-methoxyl benzotriazole (0.06 mol) Example 76 was dissolved into 150 ml of acetonitrile, 4-(3-benzisox azolyl) piperidine (0.05 mol), diisopropylethylamine (0.2 10 Preparation of N-(4-(6-cyanobenzotriazolyl)butyl)- mol) and potassium iodide (0.05 mol) were respectively 4-(3-(6-fluorobenzisoxazolyl)piperidine (I-96) added. The mixture was stirred for 10 min at ambient tem perature, and then heated and refluxed to react for 15 hours. Preparation of 1-(3-chloro The mixture was cooled down to ambient temperature and butyl)-6-cyano-1H-benzotriazole filtered. The filtrate was concentrated to produce oily prod 15 ucts, and treated by chromatography with neutral Al-O puri 6-cyano-1H-benzotriazole (15.9 g, 0.10 mol) is dissolved fied, eluted with dichloromethane/methanol mixture to pro into 100 ml of 30% wt. sodium hydroxide, 3-chlorobromobu duce 13.14 g N-(3-(6-methoxyl benzotriazolyl)propyl)-4-(3- tane (32.6 g., 0.10 mol), tetrabutyl ammonium bromide (0.8 g) benzisoxazolyl)piperidine (I-93) with a yield of 67.2%. ESI are added, and mixed for 5 min. The reaction solution is MS M+H": m/z 3912. gradually heated to 60° C., stirred for reaction for 2 hours. Post treatment was performed based on common method one Example 74 for synthesis. The solution was separated and purified by HPLC to produce 9.1 g of 1-(3-chlorobutyl)-6-cyano-1H Preparation of N-(2-(1-benzotriazolyl)ethyl)-4-(3-(6- benzotriazole, with a yield of 32.6%. fluorobenzisoxazolyl))piperidine (I-94) 25 N-(3-chlorobutyl)-6-cyano benzotriazole (0.06 mol) was dissolved into 150 ml of acetonitrile, 4-(3-(6-fluorobenzisox azolyl)) piperidine (0.05 mol), diisopropylethylamine (0.2 The method described in Example 63 was adopted to pre mol) and potassium iodide (0.05 mol) were respectively pare 1-(3-chloropropyl)-1H-benzotriazole. added. The mixture was stirred for 10 min at ambient tem 1-(3-chloropropyl)benzotriazole (0.06 mol) was dissolved perature, and then heated and refluxed to react for 15 hours. into 150 ml of acetonitrile, 4-(3-(6-fluoro benzisoxazolyl)) 30 The mixture was cooled down to ambient temperature and piperidine (0.05 mol), diisopropylethylamine (0.2 mol) and filtered. The filtrate was concentrated to produce oily prod potassium iodide (0.05 mol) were respectively added. The ucts, and treated by chromatography with neutral Al-O puri mixture was stirred for 10 min at ambient temperature, and fied, eluted with dichloromethane/methanol mixture to pro then heated and refluxed to react for 15 hours. The mixture duce 15.07 g N-(4-(6-cyano benzotriazolyl)butyl)-4-(3-(6- was cooled down to ambient temperature and filtered. The 35 benzisoxazolyl)) piperidine (I-96) with a yield of 72.1%. filtrate was concentrated to produce oily products, and treated ESI-MSM--H: m/z 418.2. by chromatography with neutral Al-O purified, eluted with dichloromethane/methanol mixture to produce 12.67 g N-(2- Example 77 (1-benzotriazolyl)ethyl)-4-(3-(6-benzisoxazolyl) piperidine (I-94) with a yield of 69.4%. ESI-MS M+H": m/z. 365.2. 40 Preparation of N-(4-(6-cyanobenzotriazolyl)butyl)- 4-(3-(6-methoxyl benzisoxazolyl)piperidine (I-97) Example 75 The method described in Example 76 was adopted to pre Preparation of N-(4-(1-benzotriazolyl)butyl)-4-(3-(6- pare N-(3-chlorobutyl)-6-cyanobenzotriazole. fluorobenzisoxazolyl))piperidine (I-95) 45 N-(3-chlorobutyl)-6-cyano benzotriazole (0.06 mol) was dissolved into 150 ml of acetonitrile, 4-(3-(6-methoxyl ben Benzotriazole (11.9 g, 0.10 mol) is dissolved into 100 ml of Zisoxazolyl)) piperidine (0.05 mol), diisopropylethylamine 30% wt. sodium hydroxide, 4-chlorobromobutane (34.3 g, (0.2 mol) and potassium iodide (0.05 mol) were respectively 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are added, added. The mixture was stirred for 10 min at ambient tem and mixed for 5 min. The reaction solution is gradually heated 50 perature, and then heated and refluxed to react for 15 hours. to 60° C., stirred for reaction for 2 hours. Then the reaction The mixture was cooled down to ambient temperature and solution was cooled down to ambient temperature, 100 ml of filtered. The filtrate was concentrated to produce oily prod dichloromethane was added for extraction and liquid separa ucts, and treated by chromatography with neutral Al-O puri tion. To the aqueous phase, 100 of dichloromethane was fied, eluted with dichloromethane/methanol mixture to pro added for extraction. Organic phases were mixed, washed 55 duce 15.01 g N-(4-(6-cyano benzotriazolyl)butyl)-4-(3-(6- with 100 ml of saturated saline. Liquid was separated, and methoxyl benzisoxazolyl)piperidine (I-97) with a yield of organic phase was evaporated to dryness to produce oily 69.8%. ESI-MSM--H: m/z 430.2. product. Oily products were separated and purified by chro matography with neutral Al-O, and eluted with dichlo Example 78 rmethane to produce 17.0 g of 1-(4-chlorobutyl)-1H-benzot 60 riazole, with a yield of 81.0%. Preparation of N-(2-(6-methoxyl benzotriazolyl) 1-(4-chlorobutyl)benzotriazole (0.06 mol) was dissolved ethoxyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine into 150 ml of acetonitrile, 4-(3-(6-fluoro benzisoxazolyl)) (I-98) piperidine (0.05 mol), diisopropylethylamine (0.2 mol) and potassium iodide (0.05 mol) were respectively added. The 65 The method described in common method four for synthe mixture was stirred for 10 min at ambient temperature, and sis was adopted to prepare N-hydroxyl-methoxyl benzotria then heated and refluxed to react for 15 hours. The mixture Zole. US 9,415,047 B2 81 82 The compound was prepared by using the methods for N-(3-chloropropyl)-6-methoxyl benzotriazole (0.06 mol) synthesis and after treatment with N-hydroxyl-6-methoxyl was dissolved into 150 ml of acetonitrile, 4-(3-benzopyrazol) benzotriazole as the material. N-(2-chloro ethoxyl-6-meth piperidine (0.05 mol), diisopropylethylamine (0.2 mol) and oxyl benzotriazole (0.06 mol) was dissolved into 150 ml of potassium iodide (0.05 mol) were respectively added. The acetonitrile, 4-(3-(6-fluorobenzisoxazolyl)) piperidine (0.05 5 mol), diisopropylethylamine (0.2 mol) and potassium iodide mixture was stirred for 10 min at ambient temperature, and (0.05 mol) were respectively added. The mixture was stirred then heated and refluxed to react for 15 hours. The mixture for 10 min at ambient temperature, and then heated and was cooled down to ambient temperature and filtered. The refluxed to react for 15 hours. The mixture was cooled down filtrate was concentrated to produce oily products, and treated to ambient temperature and filtered. The filtrate was concen by chromatography with neutral Al2O, purified, eluted with trated to produce oily products, and treated by chromatogra 10 dichloromethane/methanol mixture to produce 12.11g N-(3- phy with neutral Al-O, purified, eluted with dichlo (6-methoxyl benzotriazolyl)propyl)-4-(3-benzopyrazol)pip romethane/methanol mixture to produce 14.21 g N-(2-(6- eridine (I-101) with a yield of 66.5%. ESI-MS M+H": m/z methoxyl benzotriazolyl)ethoxyl)-4-(3-(6-fluoro 4O92. benzisoxazolyl))piperidine (I-98) with a yield of 69.1%. ESI MS M+H": m/z. 411.2. 15 Example 82 Example 79 Preparation of N-(3-(6-methoxyl benzotriazolyl) propyl)-4-(3-benzofuranyl)piperidine (I-102) Preparation of N-(2-(1-benzotriazolyl)ethoxyl)-4-(3- (6-fluorobenzisoxazolyl))piperidine (I-99) The method described in Example 73 was adopted to pre pare N-(3-chloropropyl)-6-methoxyl benzotriazole. The method described in common method four for synthe N-(3-chloropropyl)-6-methoxyl benzotriazole (0.06 mol) sis was adopted to prepare N-hydroxyl benzotriazole. was dissolved into 150 ml of acetonitrile, 4-(3-benzofuranyl) The compound was prepared by using the methods for piperidine (0.05 mol), diisopropylethylamine (0.2 mol) and synthesis and after treatment with N-hydroxyl benzotriazole 25 potassium iodide (0.05 mol) were respectively added. The as the material. N-(2-chloroethoxylbenzotriazole (0.06 mol) mixture was stirred for 10 min at ambient temperature, and was dissolved into 150 ml of acetonitrile, 4-(3-(6-fluoroben then heated and refluxed to react for 15 hours. The mixture Zisoxazolyl)) piperidine (0.05 mol), diisopropylethylamine was cooled down to ambient temperature and filtered. The (0.2 mol) and potassium iodide (0.05 mol) were respectively filtrate was concentrated to produce oily products, and treated added. The mixture was stirred for 10 min at ambient tem 30 by chromatography with neutral Al2O, purified, eluted with perature, and then heated and refluxed to react for 15 hours. dichloromethane/methanol mixture to produce 12.40 g N-(3- The mixture was cooled down to ambient temperature and (6-methoxyl benzotriazolyl)propyl)-4-(3-benzofuranyl)pip filtered. The filtrate was concentrated to produce oily prod eridine (I-102) with a yield of 68.1%. ESI-MS M+H": m/z ucts, and treated by chromatography with neutral Al-O puri 4O92. fied, eluted with dichloromethane/methanol mixture to pro 35 Known synthesis approach from current techniques could duce 12.88 g N-(2-(1-benzotriazolyl)ethoxyl)-4-(3-(6-fluoro be referred to prepare relevant compound, e.g., method benzisoxazolyl))piperidine (I-99) with a yield of 67.6%. ESI described in China patent 200810207606.7 could be used to MS M+H: m/z 381.2. prepare compound (I-103 to I-106). Example 80 40 Example 83 Preparation of N-(3-(6-methoxyl benzotriazolyl) Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- propyl)-4-(3-benzisothiazolyl)piperidine (I-100) 4-(3-chlorophenyl)piperidine (I-107)

The method described in Example 73 was adopted to pre 45 1H-benzimidazole (11.8 g., 0.10 mol) was dissolved into pare N-(3-chloropropyl)-6-methoxyl benzotriazole. 200 ml of 20% wt. sodium hydroxide, 4-chlorobromobutane N-(3-chloropropyl)-6-methoxyl benzotriazole (0.06 mol) (34.3 g, 0.20 mol) and tetrabutyl ammonium bromide (1.0 g) was dissolved into 150 ml of acetonitrile, 4-(3-benzisothiaz were added, and mixed for 5 min. The mixture was heated to olyl) piperidine (0.05 mol), diisopropylethylamine (0.2 mol) 60° C., stirred to react for 2 hours. Then the reaction solution and potassium iodide (0.05 mol) were respectively added. 50 was cooled down to ambient temperature, 100 ml of dichlo The mixture was stirred for 10 min at ambient temperature, romethane was added for extraction and liquid separation. To and then heated and refluxed to react for 15 hours. The mix the aqueous phase, 100 of dichloromethane was added for ture was cooled down to ambient temperature and filtered. extraction. Organic phases were mixed, washed with 100 ml The filtrate was concentrated to produce oily products, and of Saturated Saline. Liquid was separated, and organic phase treated by chromatography with neutral Al-O purified, 55 was evaporated to dryness to produce oily product. Oily prod eluted with dichloromethane/methanol mixture to produce ucts were separated and purified by chromatography with 13.17 g N-(3-(6-methoxyl benzotriazolyl)propyl)-4-(3-ben neutral Al-O to produce 12.5 g of 1-(4-chlorobutyl)-1H Zisothiazolyl)piperidine (I-100) with a yield of 69.1%. ESI benzimidazole, with a yield of 60.0%. MS M+H": m/z 426.2. 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) 60 was dissolved into 100 ml of acetonitrile, 3-trichlorophe Example 81 nylpiperidine (5.9 g, 0.03 mol), diisopropylethylamine (15.5 g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were Preparation of N-(3-(6-methoxyl benzotriazolyl) respectively added. The mixture was stirred for 10 min at propyl)-4-(3-benzopyrazol)piperidine (I-101) ambient temperature, and then heated and refluxed to react for 65 15 hours. The mixture was cooled down to ambient tempera The method described in Example 73 was adopted to pre ture and filtered. The filtrate was concentrated to produce oily pare N-(3-chloropropyl)-6-methoxyl benzotriazole. products, and treated by chromatography with neutral Al2O, US 9,415,047 B2 83 84 eluted with dichloromethane/methanol mixture to produce stirred for 10 min, filtered and dried to produce 5.4 g solid 7.3 g compound (I-107) with a yield of 66.4%. ESI-MS compound (II-109) with a yield of 89.0%. M+H": m/z. 368.2. Example 86 Example 84 5 Preparation of N-(4-(1H-benzotriazole-1-yl)butyl)-4- Preparation of N-(4-(1H-benzotriazole-1-yl)butyl)-4- (3-chlorophenyl)piperidine (I-108) (3-trifluoromethylphenyl)piperidine (I-110) Benzotriazole (11.9 g, 0.10 mol) is dissolved into 100 ml of 10 The method described in Example 64 was adopted to pre 30% wt. sodium hydroxide, 4-chlorobromobutane (34.3 g, pare 1-(4-chlorobutyl)-1H-benzotriazole. 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are added, 1-(4-chlorobutyl)-1H-benzotriazole (0.06 mol) was dis and mixed for 5 min. The reaction solution is gradually heated solved into 150 ml of acetonitrile, 4-(3-(3-trifluorometh to 60° C., stirred for reaction for 2 hours. Then the reaction ylphenyl) piperidine (0.05 mol), diisopropylethylamine (0.2 15 mol) and potassium iodide (0.05 mol) were respectively solution was cooled down to ambient temperature, 100 ml of added. The mixture was stirred for 10 min at ambient tem dichloromethane was added for extraction and liquid separa perature, and then heated and refluxed to react for 15 hours. tion. To the aqueous phase, 100 of dichloromethane was The mixture was cooled down to ambient temperature and added for extraction. Organic phases were mixed, washed filtered. The filtrate was concentrated to produce oily prod with 100 ml of saturated saline. Liquid was separated, and ucts, and treated by chromatography with neutral Al-O puri organic phase was evaporated to dryness to produce oily fied, eluted with dichloromethane/methanol mixture to pro product. Oily products were separated and purified by chro duce 13.6 g N-(4-(1H-benzotriazole-1-yl)butyl)-4-(3- matography with neutral Al-O, and eluted with dichlo trifluoromethylphenyl)piperidine (I-110) with a yield of rmethane to produce 17.0 g of 1-(4-chlorobutyl)-1H-benzot 67.8%. ESI-MSM--H: m/z 403.2. riazole, with a yield of 81.0%. 25 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, 3-trichlorophe Example 87 nylpiperidine (5.9 g, 0.03 mol), diisopropylethylamine (15.5 g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- respectively added. The mixture was stirred and mixed, then 30 4-(3-fluorophenyl)piperidine (I-111) heated and refluxed to react for 15 hours. The mixture was cooled down to ambient temperature and filtered. The filtrate The method described in Example 63 was adopted to pre was concentrated to produce oily products, and treated by pare 1-(4-chlorobutyl)-1H-benzimidazole. chromatography with neutral Al-O and purified, eluted with 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) dichloromethane to produce 7.8 g compound (I-108) with a 35 was dissolved into 100 ml of acetonitrile, 3-trifluorophe yield of 70.3%. ESI-MS M+H": m/z. 369.2. nylpiperidine (5.9 g, 0.03 mol), diisopropylethylamine (15.5 Compound (I-108) (5.55g, 0.015 mol) was dissolved in 50 g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were ml of ethyl acetate. Under cooling conditions of icy water respectively added. The mixture was stirred for 10 min at bath, 3 mol/L hydrogen chloride/ethyl acetate solution is ambient temperature, and then heated and refluxed to react for dripped, and the pH value is adjusted to 2. The mixture is 40 15 hours. The mixture was cooled down to ambient tempera stirred for 10 min, filtered and dried to produce 5.4 g solid ture and filtered. The filtrate was concentrated to produce oily compound (II-108) with a yield of 88.0%. products, and treated by chromatography with neutral Al-O. Example 85 eluted with dichloromethane/methanol mixture to produce 45 7.1 g compound (I-111) with a yield of 67.2%. ESI-MS Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- M+H": m/z. 352.2. 4-(3-trifluoromethylphenyl)piperidine (I-109) Example 88 The method described in Example 63 was adopted to pre pare 1-(4-chlorobutyl)-1H-benzimidazole. 50 Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- 1-(4-chlorobutyl)-1H-benzimidazole (0.06 mol) was dis 4-(2-methoxylphenyl)piperidine (I-112) solved into 150 ml of acetonitrile, 4-(3-(3-trifluorometh ylphenyl) piperidine (0.05 mol), diisopropylethylamine (0.2 mol) and potassium iodide (0.05 mol) were respectively The method described in Example 63 was adopted to pre added. The mixture was stirred for 10 min at ambient tem 55 pare 1-(4-chlorobutyl)-1H-benzimidazole. perature, and then heated and refluxed to react for 15 hours. 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) The mixture was cooled down to ambient temperature and was dissolved into 100 ml of acetonitrile, 2-methoxyphenyl filtered. The filtrate was concentrated to produce oily prod piperidine (5.7g, 0.03 mol), diisopropylethylamine (15.5 g. ucts, and treated by chromatography with neutral Al-O puri 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec fied, eluted with dichloromethane/methanol mixture to pro 60 tively added. The mixture was stirred for 10 min at ambient duce 11.0 g N-(4-(1H-benzimidazoyl-1-yl)butyl)-4-(3- temperature, and then heated and refluxed to react for 10-15 trifluoromethylphenyl)piperidine (I-109) with a yield of hours. The mixture was cooled down to ambient temperature 64.9%. ESI-MSM--H: m/z 402.2. and filtered. The filtrate was concentrated to produce oily Compound (I-109) (6.02g, 0.015 mol) was dissolved in 50 products, and treated by chromatography with neutral Al2O, ml of ethyl acetate. Under cooling conditions of icy water 65 eluted with dichloromethane/methanol mixture to produce bath, 3 mol/L hydrogen chloride/ethyl acetate solution is 6.7 g compound (I-112) with a yield of 61.3%. ESI-MS dripped, and the pH value is adjusted to 2. The mixture is M+H: m/z. 364.2. US 9,415,047 B2 85 86 Example 89 produce 14.0 g N-(4-(1H-benzotriazole-1-yl)butyl)-4-(3-tri fluoromethylphenyl)piperidine (I-114) with a yield of 64.6%. Preparation of N-(4-(6-fluoro-1H-benzotriazole-1-yl) ESI-MSM--H+: m/z. 433.2. butyl)-4-(3-trifluoromethylphenyl)piperidine (I-113) Example 91 6-fluoro-benzotriazole (15.3g, 0.10 mol) is dissolved into 100 ml of 30% wt. sodium hydroxide, 4-chlorobromobutane Preparation of N-(4-(6-cyano-1H-benzotriazole-1-yl) (34.3 g, 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are butyl)-4-(3-trifluoromethylphenyl)piperidine (I-115) added, and mixed for 5 min. The reaction solution is gradually heated to 60° C., stirred for reaction for 2 hours. Then the 10 6-cyano-benzotriazole (14.4g, 0.10 mol) is dissolved into reaction solution was cooled down to ambient temperature, 100 ml of 30% wt. sodium hydroxide, 4-chlorobromobutane 100 ml of dichloromethane was added for extraction and (34.3 g, 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are added, and mixed for 5 min. The reaction solution is gradually liquid separation. To the aqueous phase, 100 of dichlo heated to 60° C., stirred for reaction for 2 hours. Then the romethane was added for extraction. Organic phases were 15 reaction Solution was cooled down to ambient temperature, mixed, washed with 100 ml of saturated saline. Liquid was 100 ml of dichloromethane was added for extraction and separated, and organic phase was evaporated to dryness to liquid separation. To the aqueous phase, 100 of dichlo produce oily product. Oily products were separated and puri romethane was added for extraction. Organic phases were fied by chromatography with neutral Al-O, and eluted with mixed, washed with 100 ml of saturated saline. Liquid was dichlormethane to produce 17.0 g of 6-fluoro-1-(4-chlorobu separated, and organic phase was evaporated to dryness to tyl)-1H-benzotriazole, with a yield of 77.0%. produce oily product. Oily products were separated and puri 6-fluoro-1-(4-chlorobutyl)-1H-benzotriazole (0.06 mol) fied by chromatography with neutral Al-O, and eluted with was dissolved into 150 ml of acetonitrile, 4-(3-(3-trifluorom dichlormethane to produce 17.3 g of 6-cyano-1-(4-chlorobu ethylphenyl) piperidine (0.05 mol), diisopropylethylamine tyl)-1H-benzotriazole, with a yield of 74.0%. (0.2 mol) and potassium iodide (0.05 mol) were respectively 25 6-cyano-1-(4-chlorobutyl)-1H-benzotriazole (0.06 mol) added. The mixture was stirred for 10 min at ambient tem was dissolved into 150 ml of acetonitrile, 4-(3-(3-trifluorom perature, and then heated and refluxed to react for 15 hours. ethylphenyl) piperidine (0.05 mol), diisopropylethylamine The mixture was cooled down to ambient temperature and (0.2 mol) and potassium iodide (0.05 mol) were respectively filtered. The filtrate was concentrated to produce oily prod added. The mixture was stirred for 10 min at ambient tem ucts, and treated by chromatography with neutral Al-O puri 30 perature, and then heated and refluxed to react for 15 hours. fied, eluted with dichloromethane/methanol mixture to pro The mixture was cooled down to ambient temperature and duce 13.5 g N-(4-(1H-benzotriazole-1-yl)butyl)-4-(3- filtered. The filtrate was concentrated to produce oily prod trifluoromethylphenyl)piperidine (I-113) with a yield of ucts, and treated by chromatography with neutral Al-O puri fied, eluted with dichloromethane/methanol mixture to pro 64.1%. ESI-MSM--H: m/z 421.2. 35 duce 13.5 g N-(4-(1H-benzotriazole-1-yl)butyl)-4-(3- Example 90 trifluoromethylphenyl)piperidine (I-115) with a yield of 63.1%. ESI-MSM--H: m/z 427.2. Preparation of N-(4-(6-methoxyl-1H-benzotriazole Example 92 1-yl)butyl)-4-(3-trifluoromethylphenyl)piperidine 40 (I-114) Preparation of N-(4-(1H-benzotriazole-1-yl)pro poxyl)-4-(3-trifluoromethylphenyl)piperidine (I-116) 6-methoxyl-benzotriazole (14.9 g, 0.10 mol) is dissolved into 100 ml of 30% wt. sodium hydroxide, 4-chlorobromobu Preparation of N-(2-chloro propoxyl)benzotriazole tane (34.3g, 0.20 mol), tetrabutyl ammonium bromide (0.8 g) 45 are added, and mixed for 5 min. The reaction solution is Substituted 1-hydroxyl benzotriazole (0.01 mol) was dis gradually heated to 60° C., stirred for reaction for 2 hours. solved in 10 ml of NMP solid paraffin mixture containing Then the reaction solution was cooled down to ambient tem 50% (w/w) hydrogen and oxygen was added in different perature, 100 ml of dichloromethane was added for extraction times, stirred to react for 0.5 h. Meanwhile, 3-bromochloro and liquid separation. To the aqueous phase, 100 of dichlo 50 propane (0.015 mol) was dissolved in 5 ml of NMP and added romethane was added for extraction. Organic phases were into the above said solution, and stirred to react for 12 h. mixed, washed with 100 ml of saturated saline. Liquid was Reaction solution was poured into 50 ml of water, extracted separated, and organic phase was evaporated to dryness to with ethyl acetate (3x50 mL). Organic phases were mixed and produce oily product. Oily products were separated and puri washed with 30 ml of water. Anhydrous magnesium sulfate fied by chromatography with neutral Al-O, and eluted with 55 was added to dry organic phase, filtered, with solvent evapo dichlormethane to produce 17.9 g of 6-methoxyl-1-(4-chlo rated. Oily products were analyzed by chromatography with robutyl)-1H-benzotriazole, with a yield of 75.0%. neutral Al-O, or separated and purified by using HPLC to 6-methoxyl-1-(4-chlorobutyl)-1H-benzotriazole (0.06 prepare 1-(3-chloropropoxyl)benzotriazole, with a yield of mol) was dissolved into 150 ml of acetonitrile, 4-(3-(3-trif 75.0%-85.0%. luoromethylphenyl) piperidine (0.05 mol), diisopropylethy 60 1-(3-chloropropoxyl)benzotriazole (0.06 mol) was dis lamine (0.2 mol) and potassium iodide (0.05 mol) were solved into 150 ml of acetonitrile, 4-(3-(6-fluorobenzisox respectively added. The mixture was stirred for 10 min at azolyl)) piperidine (0.05 mol), diisopropylethylamine (0.2 ambient temperature, and then heated and refluxed to react for mol) and potassium iodide (0.05 mol) were respectively 15 hours. The mixture was cooled down to ambient tempera added. The mixture was stirred for 10 min at ambient tem ture and filtered. The filtrate was concentrated to produce oily 65 perature, and then heated and refluxed to react for 15 hours. products, and treated by chromatography with neutral Al2O, The mixture was cooled down to ambient temperature and purified, eluted with dichloromethane/methanol mixture to filtered. The filtrate was concentrated to produce oily prod US 9,415,047 B2 87 88 ucts, and treated by chromatography with neutral Al-O puri filtered. The filtrate was concentrated to produce oily prod fied, eluted with dichloromethane/methanol mixture to pro ucts, and treated by chromatography with neutral Al-O. duce 13.2 g N-(4-(1H-benzotriazole-1-yl)propoxyl)-4-(3- eluted with dichloromethane/methanol mixture to produce trifluoromethylphenyl)piperidine (I-116) with a yield of 6.0 g compound (I-118) with a yield of 61.6%. ESI-MS 65.3%. ESI-MSM--H: m/z 405.2. 5 M+H": m/z 324.2. Example 93 Example 95 Preparation of N-(4-(1H-benzimidazole-1-yl)pro poxyl)-4-(3-trifluoromethylphenyl)piperidine (I-117) 10 Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- 4-(4-pyridyl)piperidine (I-119) Preparation of N-(2-chloro propoxyl)benzimidazole The method described in Example 94 was adopted to pre Substituted 1-hydroxylbenzimidazole (0.01 mol) was dis pare 1-(4-chlorobutyl)-1H-benzimidazole. solved in 10 ml of NMP solid paraffin mixture containing 15 50% (w/w) hydrogen and oxygen was added in different 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) times, stirred to react for 0.5 h. Meanwhile, 3-bromochloro was dissolved into 100 ml of acetonitrile, 4-(4-pyridyl) pip propane (0.015 mol) was dissolved in 5 ml of NMP and added eridine (4.9 g, 0.03 mol), diisopropylethylamine (15.5g, 0.12 into the above said solution, and stirred to react for 12 h. mol) and potassium iodide (5.0g, 0.03 mol) were respectively Reaction solution was poured into 50 ml of water, extracted added. The mixture was stirred for 10 min at ambient tem with ethyl acetate (3x50 mL). Organic phases were mixed and perature, and then heated and refluxed to react for 20 hours. washed with 30 ml of water. Anhydrous magnesium sulfate The mixture was cooled down to ambient temperature and was added to dry organic phase, filtered, with solvent evapo filtered. The filtrate was concentrated to produce oily prod rated. Oily products were analyzed by chromatography with ucts, and treated by chromatography with neutral Al-O. neutral Al-O, or separated and purified by using HPLC to 25 eluted with dichloromethane/methanol mixture to produce prepare 1-(3-chloropropoxyl)benzimidazole, with a yield of 6.3 g compound (I-119) with a yield of 62.1%. ESI-MS 75.0%. M+H: m/z 335.2. 1-(3-chloropropoxyl)benzimidazole (0.06 mol) was dis solved into 150 ml of acetonitrile, 4-(3-(6-fluorobenzisox Example 96 azolyl)) piperidine (0.05 mol), diisopropylethylamine (0.2 30 mol) and potassium iodide (0.05 mol) were respectively added. The mixture was stirred for 10 min at ambient tem Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- perature, and then heated and refluxed to react for 15 hours. 4-(2-pyrimidinyl)piperidine (I-120) The mixture was cooled down to ambient temperature and filtered. The filtrate was concentrated to produce oily prod 35 The method described in Example 94 was adopted to pre ucts, and treated by chromatography with neutral Al-O puri pare 1-(4-chlorobutyl)-1H-benzimidazole. fied, eluted with dichloromethane/methanol mixture to pro 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) duce N-(4-(1H-benzimidazole-1-yl)propoxyl)-4-(3- was dissolved into 100 ml of acetonitrile, 4-(2-pyrimidinyl) trifluoromethylphenyl)piperidine (I-117) with a yield of piperidine (4.9 g, 0.03 mol), diisopropylethylamine (15.5 g. 67.1%. ESI-MSM--H: m/z 404.2. 40 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec tively added. The mixture was stirred for 10 min at ambient Example 94 temperature, and then heated and refluxed to react for 20 hours. The mixture was cooled down to ambient temperature Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- and filtered. The filtrate was concentrated to produce oily 4-(2-furyl)piperidine (I-118) 45 products, and treated by chromatography with neutral Al2O, eluted with dichloromethane/methanol mixture to produce 1H-benzimidazole (11.8 g., 0.10 mol) was dissolved into 6.1 g compound (I-120) with a yield of 60.1%. ESI-MS 200 ml of 20% wt. sodium hydroxide, 4-chlorobromobutane M+H": m/z 336.2. (34.3 g, 0.20 mol) and tetrabutyl ammonium bromide (1.0 g) were added, and mixed for 5 min. The mixture was heated to 50 60° C., stirred to react for 2 hours. Then the reaction solution Example 97 was cooled down to ambient temperature, 100 ml of dichlo romethane was added for extraction and liquid separation. To Preparation of N-(4-(1H-benzotriazole-1-yl)butyl)-4- the aqueous phase, 100 of dichloromethane was added for (4-cyclohexyl)piperidine (I-121) extraction. Organic phases were mixed, washed with 100 ml 55 of Saturated Saline. Liquid was separated, and organic phase was evaporated to dryness to produce oily product. Oily prod The method described in Example 64 was adopted to pre ucts were separated and purified by chromatography with pare 1-(4-chlorobutyl)-1H-benzotriazole. neutral Al-O to produce 12.5 g of 1-(4-chlorobutyl)-1H 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) benzimidazole, with a yield of 60.0%. 60 was dissolved into 100 ml of acetonitrile, 4-(1-cyclohexyl) 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) piperidine (5.1 g, 0.03 mol), diisopropylethylamine (15.5 g. was dissolved into 100 ml of acetonitrile, 4-(2-furyl) piperi 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec dine (4.6 g., 0.03 mol), diisopropylethylamine (15.5 g., 0.12 tively added. The mixture was stirred and mixed, then heated mol) and potassium iodide (5.0 g, 0.03 mol) were respectively and refluxed to react for 20 hours. The mixture was cooled added. The mixture was stirred for 10 min at ambient tem 65 down to ambient temperature and filtered. The filtrate was perature, and then heated and refluxed to react for 20 hours. concentrated to produce oily products, and treated by chro The mixture was cooled down to ambient temperature and matography with neutral Al-O and purified, eluted with US 9,415,047 B2 89 90 dichloromethane to produce 6.5 g compound (I-121) with a ture and filtered. The filtrate was concentrated to produce oily yield of 63.7%. ESI-MS M+H: m/z 341.3. products, and treated by chromatography with neutral Al-O. eluted with dichloromethane/methanol mixture to produce Example 98 7.3 g compound (I-124) with a yield of 66.4%. ESI-MS M+H": m/z. 368.2. Preparation of N-(4-(1H-benzotriazole-1-yl)butyl)-4- (1-naphthyl)piperidine (I-122) Example 101 The method described in Example 64 was adopted to pre Preparation of N-(4-(1H-benzotriazole-1-yl)butyl)-4- pare 1-(4-chlorobutyl)-1H-benzotriazole. 10 (3-chlorophenyl)piperidine (I-125) 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, 4-(1-naphthyl)pip Benzotriazole (11.9 g, 0.10 mol) is dissolved into 100 ml of eridine (6.4g, 0.03 mol), diisopropylethylamine (15.5g, 0.12 30% wt. sodium hydroxide, 4-chlorobromobutane (34.3 g, mol) and potassium iodide (5.0 g, 0.03 mol) were respectively 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are added, added. The mixture was stirred and mixed, then heated and 15 and mixed for 5 min. The reaction solution is gradually heated refluxed to react for 20 hours. The mixture was cooled down to 60° C., stirred for reaction for 2 hours. Then the reaction to ambient temperature and filtered. The filtrate was concen solution was cooled down to ambient temperature, 100 ml of trated to produce oily products, and treated by chromatogra dichloromethane was added for extraction and liquid separa phy with neutral Al-O, and purified, eluted with dichlo tion. To the aqueous phase, 100 of dichloromethane was romethane to produce 6.9 g compound (I-122) with a yield of added for extraction. Organic phases were mixed, washed 60.1%. ESI-MSM--H: m/z 385.3. with 100 ml of saturated saline. Liquid was separated, and organic phase was evaporated to dryness to produce oily Example 99 product. Oily products were separated and purified by chro matography with neutral Al-O, and eluted with dichlo Preparation of N-(4-(1H-benzotriazole-1-yl)butyl)-4- 25 rmethane to produce 17.0 g of 1-(4-chlorobutyl)-1H-benzot (2-quinoxalinyl)piperidine (I-123) riazole, with a yield of 81.0%. 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) The method described in Example 64 was adopted to pre was dissolved into 100 ml of acetonitrile, 3-trichlorophe pare 1-(4-chlorobutyl)-1H-benzotriazole. nylpiperidine (5.9 g, 0.03 mol), diisopropylethylamine (15.5 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) 30 g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were was dissolved into 100 ml of acetonitrile, 4-(2-quinoxalinyl) respectively added. The mixture was stirred and mixed, then piperidine (6.4g, 0.03 mol), diisopropylethylamine (15.5 g. heated and refluxed to react for 15 hours. The mixture was 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec cooled down to ambient temperature and filtered. The filtrate tively added. The mixture was stirred and mixed, then heated was concentrated to produce oily products, and treated by and refluxed to react for 20 hours. The mixture was cooled 35 chromatography with neutral Al-O and purified, eluted with down to ambient temperature and filtered. The filtrate was dichloromethane to produce 7.8 g compound (I-125) with a concentrated to produce oily products, and treated by chro yield of 70.3%. ESI-MS M+H": m/z. 369.2. matography with neutral Al-O and purified, eluted with Compound (I-124) (5.55g, 0.015 mol) was dissolved in 50 dichloromethane to produce 7.3 g compound (I-123) with a ml of ethyl acetate. Under cooling conditions of icy water yield of 62.7%. ESI-MS M+H: m/z 387.2. 40 bath, 3 mol/L hydrogen chloride/ethyl acetate solution is dripped, and the pH value is adjusted to 2. The mixture is Example 100 stirred for 10 min, filtered and dried to produce 5.4 g solid compound (II-125) with a yield of 88.0%. Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- 4-(3-chlorophenyl)piperidine (I-124) 45 Example 102 1H-benzimidazole (11.8 g., 0.10 mol) was dissolved into Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- 200 ml of 20% wt. sodium hydroxide, 4-chlorobromobutane 4-(3-trifluoromethylphenyl)piperidine (I-126) (34.3 g, 0.20 mol) and tetrabutyl ammonium bromide (1.0 g) were added, and mixed for 5 min. The mixture was heated to 50 The method described in Example 100 was adopted to 60° C., stirred to react for 2 hours. Then the reaction solution prepare 1-(4-chlorobutyl)-1H-benzimidazole. was cooled down to ambient temperature, 100 ml of dichlo 1-(4-chlorobutyl)-1H-benzimidazole (0.06 mol) was dis romethane was added for extraction and liquid separation. To solved into 150 ml of acetonitrile, 4-(3-(3-trifluorometh the aqueous phase, 100 of dichloromethane was added for ylphenyl) piperidine (0.05 mol), diisopropylethylamine (0.2 extraction. Organic phases were mixed, washed with 100 ml 55 mol) and potassium iodide (0.05 mol) were respectively of Saturated Saline. Liquid was separated, and organic phase added. The mixture was stirred for 10 min at ambient tem was evaporated to dryness to produce oily product. Oily prod perature, and then heated and refluxed to react for 15 hours. ucts were separated and purified by chromatography with The mixture was cooled down to ambient temperature and neutral Al-O to produce 12.5 g of 1-(4-chlorobutyl)-1H filtered. The filtrate was concentrated to produce oily prod benzimidazole, with a yield of 60.0%. 60 ucts, and treated by chromatography with neutral Al-O puri 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) fied, eluted with dichloromethane/methanol mixture to pro was dissolved into 100 ml of acetonitrile, 3-trichlorophe duce 11.0 g N-(4-(1H-benzimidazoyl-1-yl)butyl)-4-(3- nylpiperidine (5.9 g, 0.03 mol), diisopropylethylamine (15.5 trifluoromethylphenyl)piperidine (I-126) with a yield of g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were 64.9%. ESI-MSM--H: m/z 402.2. respectively added. The mixture was stirred for 10 min at 65 Compound (I-126) (6.02g, 0.015 mol) was dissolved in 50 ambient temperature, and then heated and refluxed to react for ml of ethyl acetate. Under cooling conditions of icy water 15 hours. The mixture was cooled down to ambient tempera bath, 3 mol/L hydrogen chloride/ethyl acetate solution is US 9,415,047 B2 91 92 dripped, and the pH value is adjusted to 2. The mixture is Example 106 stirred for 10 min, filtered and dried to produce 5.4 g solid compound (II-126) with a yield of 89.0%. Preparation of N-(4-(6-fluoro-1H-benzotriazole-1-yl) butyl)-4-(3-trifluoromethylphenyl)piperidine (I-130) Example 103 6-fluoro-benzotriazole (15.3 g, 0.10 mol) is dissolved into Preparation of N-(4-(1H-benzotriazole-1-yl)butyl)-4- 100 ml of 30% wt. sodium hydroxide, 4-chlorobromobutane (3-trifluoromethylphenyl)piperidine (I-127) (34.3 g, 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are 10 added, and mixed for 5 min. The reaction solution is gradually The method described in Example 101 was adopted to heated to 60° C., stirred for reaction for 2 hours. Then the prepare 1-(4-chlorobutyl)-1H-benzotriazole. reaction Solution was cooled down to ambient temperature, 1-(4-chlorobutyl)-1H-benzotriazole (0.06 mol) was dis 100 ml of dichloromethane was added for extraction and solved into 150 ml of acetonitrile, 4-(3-(3-trifluorometh liquid separation. To the aqueous phase, 100 of dichlo ylphenyl)piperidine (0.05 mol), diisopropylethylamine (25.8 15 romethane was added for extraction. Organic phases were g, 0.2 mol) and potassium iodide (0.05 mol) were respectively mixed, washed with 100 ml of saturated saline. Liquid was added. The mixture was stirred for 10 min at ambient tem separated, and organic phase was evaporated to dryness to perature, and then heated and refluxed to react for 15 hours. produce oily product. Oily products were separated and puri The mixture was cooled down to ambient temperature and fied by chromatography with neutral Al-O, and eluted with filtered. The filtrate was concentrated to produce oily prod dichlormethane to produce 17.0 g of 6-fluoro-1-(4-chlorobu ucts, and treated by chromatography with neutral Al-O puri tyl)-1H-benzotriazole, with a yield of 77.0%. fied, eluted with dichloromethane/methanol mixture to pro 6-fluoro-1-(4-chlorobutyl)-1H-benzotriazole (0.06 mol) duce 13.6 g N-(4-(1H-benzotriazole-1-yl)butyl)-4-(3- was dissolved into 150 ml of acetonitrile, 4-(3-(3-trifluorom trifluoromethylphenyl)piperidine (I-127) with a yield of ethylphenyl) piperidine (0.05 mol), diisopropylethylamine 67.8%. ESI-MSM--H: m/z 403.2. 25 (0.2 mol) and potassium iodide (0.05 mol) were respectively added. The mixture was stirred for 10 min at ambient tem Example 104 perature, and then heated and refluxed to react for 15 hours. The mixture was cooled down to ambient temperature and Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- filtered. The filtrate was concentrated to produce oily prod 4-(3-fluorophenyl)piperidine (I-128) 30 ucts, and treated by chromatography with neutral Al-O puri fied, eluted with dichloromethane/methanol mixture to pro The method described in Example 100 was adopted to duce 13.5 g N-(4-(1H-benzotriazole-1-yl)butyl)-4-(3- prepare 1-(4-chlorobutyl)-1H-benzimidazole. trifluoromethylphenyl)piperidine (I-130) with a yield of 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) 35 64.1%. ESI-MSM--H: m/z 421.2. was dissolved into 100 ml of acetonitrile, 3-trifluorophe nylpiperidine (5.9 g, 0.03 mol), diisopropylethylamine (15.5 Example 107 g, 0.12 mol) and potassium iodide (5.0 g, 0.03 mol) were respectively added. The mixture was stirred for 10 min at Preparation of N-(4-(6-methoxyl-1H-benzotriazole ambient temperature, and then heated and refluxed to react for 40 1-yl)butyl)-4-(3-trifluoromethylphenyl)piperidine 15 hours. The mixture was cooled down to ambient tempera (I-131) ture and filtered. The filtrate was concentrated to produce oily products, and treated by chromatography with neutral Al2O, 6-methoxyl-benzotriazole (14.9 g, 0.10 mol) is dissolved eluted with dichloromethane/methanol mixture to produce into 100 ml of 30% wt. sodium hydroxide, 4-chlorobromobu 7.1 g compound (I-128) with a yield of 67.2%. ESI-MS 45 tane (34.3g, 0.20 mol), tetrabutyl ammonium bromide (0.8 g) M+H": m/z. 352.2. are added, and mixed for 5 min. The reaction solution is gradually heated to 60° C., stirred for reaction for 2 hours. Example 105 Then the reaction solution was cooled down to ambient tem perature, 100 ml of dichloromethane was added for extraction 50 and liquid separation. To the aqueous phase, 100 of dichlo Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- romethane was added for extraction. Organic phases were 4-(2-methoxylphenyl)piperidine (I-119) mixed, washed with 100 ml of saturated saline. Liquid was separated, and organic phase was evaporated to dryness to The method described in Example 100 was adopted to produce oily product. Oily products were separated and puri prepare 1-(4-chlorobutyl)-1H-benzimidazole. 55 fied by chromatography with neutral Al-O, and eluted with 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) dichlormethane to produce 17.9 g of 6-methoxyl-1-(4-chlo was dissolved into 100 ml of acetonitrile, 2-methoxyphenyl robutyl)-1H-benzotriazole, with a yield of 75.0%. piperidine (5.7g, 0.03 mol), diisopropylethylamine (15.5 g. 6-methoxyl-1-(4-chlorobutyl)-1H-benzotriazole (0.06 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec mol) was dissolved into 150 ml of acetonitrile, 4-(3-(3-trif tively added. The mixture was stirred for 10 min at ambient 60 luoromethylphenyl) piperidine (0.05 mol), diisopropylethy temperature, and then heated and refluxed to react for 10-15 lamine (0.2 mol) and potassium iodide (0.05 mol) were hours. The mixture was cooled down to ambient temperature respectively added. The mixture was stirred for 10 min at and filtered. The filtrate was concentrated to produce oily ambient temperature, and then heated and refluxed to react for products, and treated by chromatography with neutral Al2O, 15 hours. The mixture was cooled down to ambient tempera eluted with dichloromethane/methanol mixture to produce 65 ture and filtered. The filtrate was concentrated to produce oily 6.7 g compound (I-129) with a yield of 61.3%. ESI-MS products, and treated by chromatography with neutral Al2O, M+H: m/z. 364.2. purified, eluted with dichloromethane/methanol mixture to US 9,415,047 B2 93 94 produce 14.0 g N-(4-(1H-benzotriazole-1-yl)butyl)-4-(3-tri ucts, and treated by chromatography with neutral Al-O puri fluoromethylphenyl)piperidine (I-131) with a yield of 64.6%. fied, eluted with dichloromethane/methanol mixture to pro ESI-MSM--H: m/z. 433.2. duce 13.2 g N-(4-(1H-benzotriazole-1-yl)propoxyl)-4-(3- trifluoromethylphenyl)piperidine (I-133) with a yield of Example 108 65.3%. ESI-MSM--H: m/z 405.2. Preparation of N-(4-(6-cyano-1H-benzotriazole-1-yl) Example 110 butyl)-4-(3-trifluoromethylphenyl)piperidine (I-132) Preparation of N-(4-(1H-benzimidazole-1-yl)pro 6-cyano-benzotriazole (14.4g, 0.10 mol) is dissolved into 10 poxyl)-4-(3-trifluoromethylphenyl)piperidine (I-134) 100 ml of 30% wt. sodium hydroxide, 4-chlorobromobutane (34.3 g, 0.20 mol), tetrabutyl ammonium bromide (0.8 g) are Preparation of N-(2-chloro propoxyl)benzimidazole added, and mixed for 5 min. The reaction solution is gradually heated to 60° C., stirred for reaction for 2 hours. Then the Substituted 1-hydroxyl benzimidazole (0.01 mol) was dis reaction solution was cooled down to ambient temperature, 15 solved in 10 ml of NMP solid paraffin mixture containing 100 ml of dichloromethane was added for extraction and 50% (w/w) hydrogen and oxygen was added in different liquid separation. To the aqueous phase, 100 of dichlo times, stirred to react for 0.5 h. Meanwhile, 3-bromochloro romethane was added for extraction. Organic phases were propane (0.015 mol) was dissolved in 5 ml of NMP and added mixed, washed with 100 ml of saturated saline. Liquid was into the above said solution, and stirred to react for 12 h. separated, and organic phase was evaporated to dryness to Reaction solution was poured into 50 ml of water, extracted produce oily product. Oily products were separated and puri with ethyl acetate (3x50 mL). Organic phases were mixed and fied by chromatography with neutral Al-O, and eluted with washed with 30 ml of water. Anhydrous magnesium sulfate dichlormethane to produce 17.3 g of 6-cyano-1-(4-chlorobu was added to dry organic phase, filtered, with solvent evapo tyl)-1H-benzotriazole, with a yield of 74.0%. rated. Oily products were analyzed by chromatography with 6-cyano-1-(4-chlorobutyl)-1H-benzotriazole (0.06 mol) 25 neutral Al-O, or separated and purified by using HPLC to was dissolved into 150 ml of acetonitrile, 4-(3-(3-trifluorom prepare 1-(3-chloropropoxyl)benzimidazole, with a yield of ethylphenyl) piperidine (0.05 mol), diisopropylethylamine 75.0%. (0.2 mol) and potassium iodide (0.05 mol) were respectively 1-(3-chloropropoxyl)benzimidazole (0.06 mol) was dis added. The mixture was stirred for 10 min at ambient tem solved into 150 ml of acetonitrile, 4-(3-(6-fluorobenzisox perature, and then heated and refluxed to react for 15 hours. 30 azolyl)) piperidine (0.05 mol), diisopropylethylamine (0.2 The mixture was cooled down to ambient temperature and mol) and potassium iodide (0.05 mol) were respectively filtered. The filtrate was concentrated to produce oily prod added. The mixture was stirred for 10 min at ambient tem ucts, and treated by chromatography with neutral Al-O puri perature, and then heated and refluxed to react for 15 hours. fied, eluted with dichloromethane/methanol mixture to pro The mixture was cooled down to ambient temperature and duce 13.5 g N-(4-(1H-benzotriazole-1-yl)butyl)-4-(3- 35 filtered. The filtrate was concentrated to produce oily prod trifluoromethylphenyl)piperidine (I-132) with a yield of ucts, and treated by chromatography with neutral Al-O puri 63.1%. ESI-MSM--H: m/z 427.2. fied, eluted with dichloromethane/methanol mixture to pro duce 13.6 g N-(4-(1H-benzimidazole-1-yl)propoxyl)-4-(3- Example 109 trifluoromethylphenyl)piperidine (I-134) with a yield of 40 67.1%. ESI-MSM--H: m/z 404.2. Preparation of N-(4-(1H-benzotriazole-1-yl)pro poxyl)-4-(3-trifluoromethylphenyl)piperidine (I-133) Example 111 Preparation of N-(2-chloro propoxyl)benzotriazole Preparation of N-(3-(1H-benzotriazole-1-yl)propyl)- 45 4-(3-(6-methylbenzisoxazolyl))piperidine (I-135) Substituted 1-hydroxyl benzotriazole (0.01 mol) was dis solved in 10 ml of NMP solid paraffin mixture containing 1-(3-chloropropyl)-1H-benzotriazole (11.7 g., 0.06 mol) 50% (w/w) hydrogen and oxygen was added in different was dissolved into 150 ml of acetonitrile, 6-methyl-3-(piperi times, stirred to react for 0.5 h. Meanwhile, 3-bromochloro dine-4-yl)benzisoxazole (10.8 g., 0.05 mol), diisopropylethy propane (0.015 mol) was dissolved in 5 ml of NMP and added 50 lamine (25.8 g., 0.02 mol) and potassium iodide (8.3 g, 0.05 into the above said solution, and stirred to react for 12 h. mol) were respectively added. The mixture was stirred and Reaction solution was poured into 50 ml of water, extracted mixed for 10 min at ambient temperature, then heated and with ethyl acetate (3x50 mL). Organic phases were mixed and refluxed to react for 15 hours. After treatment according to washed with 30 ml of water. Anhydrous magnesium sulfate common method three for synthesis produced 12.4 g com was added to dry organic phase, filtered, with solvent evapo 55 pound (I-135), with a yield of 66.1%. ESI-MSM--HI": m/z. rated. Oily products were analyzed by chromatography with 3762. neutral Al-O, or separated and purified by using HPLC to prepare 1-(3-chloropropoxyl)benzotriazole, with a yield of Example 112 75.0%-85.0%. 1-(3-chloropropoxyl)benzotriazole (0.06 mol) was dis 60 Preparation of N-(3-(1H-benzotriazole-1-yl)propyl)- solved into 150 ml of acetonitrile, 4-(3-(6-fluorobenzisox 4-(3-(6-methylbenzisoxazolyl))piperidine (I-136) azolyl)) piperidine (0.05 mol), diisopropylethylamine (0.2 mol) and potassium iodide (0.05 mol) were respectively 1-(3-chloropropyl)-1H-benzotriazole (11.7 g., 0.06 mol) added. The mixture was stirred for 10 min at ambient tem was dissolved into 150 ml of acetonitrile, 6-methoxyl-3-(pi perature, and then heated and refluxed to react for 15 hours. 65 peridine-4-yl)benzisoxazole (11.6 g., 0.05 mol), diisopropyl The mixture was cooled down to ambient temperature and ethylamine (25.8 g., 0.02 mol) and potassium iodide (8.3 g, filtered. The filtrate was concentrated to produce oily prod 0.05 mol) were respectively added. The mixture was stirred US 9,415,047 B2 95 96 and mixed for 10 minatambient temperature, then heated and (4.15g, 0.025 mol) were respectively added. The mixture was refluxed to react for 15 hours. After treatment according to stirred and mixed for 10 min at ambient temperature, then common method three for synthesis produced 13.3 g com heated and refluxed to react for 16 hours. After treatment pound (I-136), with a yield of 67.7%. ESI-MSM--HI": m/z. according to common method three for synthesis produced 392.2. 8.1 g compound (I-138), with a yield of 65.2%. ESI-MS M+H: m/z. 414.1. Example 113 Example 115 Preparation of N-(3-(6-fluoro-1H-benzotriazole-1-yl) propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine 10 Preparation of N-(3-(6-methyl-1H-benzotriazole-1- (I-137) yl)propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine (I-139) Preparation of 1-(3-chloro propyl)-6-fluoro-1H-benzotriazole Preparation of 1-(3-chloro 15 propyl)-6-methyl-1H-benzotriazole 6-fluoro-1H-benzotriazole (13.7g, 0.10 mol) is dissolved into 100 ml of 30% wt. sodium hydroxide, 3-chlorobro 6-methyl-1H-benzotriazole (13.3 g, 0.10 mol) is dissolved mopropane (31.4g, 0.10 mol), tetrabutyl ammonium bromide into 100 ml of 30% wt. sodium hydroxide, 3-chlorobro (0.8 g) are added, and mixed for 5 min. The reaction solution mopropane (31.4g, 0.10 mol), tetrabutyl ammonium bromide is gradually heated to 60°C., stirred for reaction for 2 hours. (0.8 g) are added, and mixed for 5 min. The reaction solution Post treatment was performed based on common method two is gradually heated to 60°C., stirred for reaction for 2 hours. for synthesis. The solution was separated and purified by Post treatment was performed based on common method two HPLC to produce 6.9 g of 1-(3-chloropropyl)-6-fluoro-1H for synthesis. The solution was separated and purified by benzotriazole, with a yield of 32.3%. HPLC to produce 7.2 g of 1-(3-chloropropyl)-6-methyl-1H 25 benzotriazole, with a yield of 34.3%. Preparation of N-(3-(6-fluoro-1H-benzotriazole-1-yl) propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine Preparation of N-(3-(6-methyl-1H-benzotriazole-1- (I-137) yl)propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine (I-139) 1-(3-chloropropyl)-6-fluoro-1H-benzotriazole (6.41 g, 30 0.03 mol) was dissolved into 150 ml of acetonitrile, 6-fluoro 1-(3-chloropropyl)-6-methyl-1H-benzotriazole (6.29 g, 3-(piperidine-4-yl)benzisoxazole (5.5 g., 0.025 mol), diiso 0.03 mol) was dissolved into 150 ml of acetonitrile, 6-fluoro propylethylamine (12.9 g, 0.1 mol) and potassium iodide 3-(piperidine-4-yl)benzisoxazole (5.5 g., 0.025 mol), diiso (4.15g, 0.025 mol) were respectively added. The mixture was propylethylamine (12.9 g, 0.1 mol) and potassium iodide stirred and mixed for 10 min at ambient temperature, then 35 (4.15g, 0.025 mol) were respectively added. The mixture was heated and refluxed to react for 16 hours. After treatment stirred and mixed for 10 min at ambient temperature, then according to common method three for synthesis produced heated and refluxed to react for 16 hours. After treatment 8.3 g compound (I-137), with a yield of 69.6%. ESI-MS according to common method three for synthesis produced M+H": m/z 398.2. 8.5 g compound (I-139), with a yield of 71.9%. ESI-MS 40 M+H: m/z 394.2. Example 114 Example 116 Preparation of N-(3-(6-chloro-1H-benzotriazole-1- yl)propyl)-4-(3-(6-fluorobenzisoxazolyl)piperidine Preparation of N-(3-(6-methoxyl-1H-benzotriazole (I-138) 45 1-yl)propyl)-4-(3-(6-fluorobenzisoxazolyl))piperi dine (I-140) Preparation of 1-(3-chloro propyl)-6-chloro-1H-benzotriazole Preparation of N-(3-chloro propyl)-6-methoxyl-benzotriazole 6-chloro-1H-benzotriazole (15.4g, 0.10 mol) is dissolved 50 into 100 ml of 30% wt. sodium hydroxide, 3-chlorobro 6-methoxyl-1H-benzotriazole (14.9 g, 0.10 mol) is dis mopropane (31.4g, 0.10 mol), tetrabutyl ammonium bromide solved into 100 ml of 30% wt. sodium hydroxide, 3-chloro (0.8 g) are added, and mixed for 5 min. The reaction solution bromopropane (31.4g, 0.10 mol), tetrabutyl ammonium bro is gradually heated to 60°C., stirred for reaction for 2 hours. mide (0.8 g) are added, and mixed for 5 min. The reaction Post treatment was performed based on common method one 55 solution is gradually heated to 60°C., stirred for reaction for for synthesis. The solution was separated and purified by 2 hours. Post treatment was performed based on common HPLC to produce 7.3 g of 1-(3-chloropropyl)-6-chloro-1H method two for synthesis. The Solution was separated and benzotriazole, with a yield of 31.7%. purified by HPLC to produce 7.7 g of N-(3-chloropropyl)-6- methoxyl-benzotriazole, with a yield of 34.1%. Preparation of N-(3-(6-chloro-1H-benzotriazole-1- 60 yl)propyl)-4-(3-(6-fluorobenzisoxazolyl) piperidine Preparation of N-(3-(6-methoxyl-1H-benzotriazole (I-138) 1-yl)propyl)-4-(3-(6-fluorobenzisoxazolyl))piperi dine (I-140) 1-(3-chloropropyl)-6-chloro-1H-benzotriazole (6.90 g, 0.03 mol) was dissolved into 150 ml of acetonitrile, 6-fluoro 65 1-(3-chloropropyl)-6-methoxyl-1H-benzotriazole (6.77 g. 3-(piperidine-4-yl)benzisoxazole (5.5 g., 0.025 mol), diiso 0.03 mol) was dissolved into 150 ml of acetonitrile, 6-fluoro propylethylamine (12.9 g, 0.1 mol) and potassium iodide 3-(piperidine-4-yl)benzisoxazole (5.5 g., 0.025 mol), diiso US 9,415,047 B2 97 98 propylethylamine (12.9 g, 0.1 mol) and potassium iodide 0.10 mol), tetrabutyl ammonium bromide (0.8 g) are added, (4.15g, 0.025 mol) were respectively added. The mixture was and mixed for 5 min. The reaction solution is gradually heated stirred and mixed for 10 min at ambient temperature, then to 60° C., stirred for reaction for 2 hours. Then the reaction heated and refluxed to react for 16 hours. After treatment solution was cooled down to ambient temperature, 100 ml of according to common method three for synthesis produced dichloromethane was added for extraction and liquid separa 8.6 g compound (I-140), with a yield of 70%. ESI-MS tion. To the aqueous phase, 100 of dichloromethane was M+H: m/z 410.2. added for extraction. Organic phases were mixed, washed Example 117 with 100 ml of saturated saline. Liquid was separated, and organic phase was evaporated to dryness to produce oily Preparation of N-(3-(6-formoxyl-1H-benzotriazole 10 product. Oily products were separated and purified by chro 1-yl)propyl)-4-(3-(6-fluorobenzisoxazolyl))piperi matography with neutral Al-O, and eluted and separated dine (I-141) with dichlormethane to produce 16.0 g of 1-(3-chloropropyl)- 1H-benzotriazole, with a yield of 82.0%. Preparation of 1-(3-chloro 1-(3-chloropropyl)benzotriazole (0.06 mol) was dissolved propyl)-6-formoxyl-1H-benzotriazole 15 into 150 ml of acetonitrile, 4-(3-(6-fluoro benzisoxazolyl)) piperidine (0.05 mol), diisopropylethylamine (0.2 mol) and 6-formoxyl-1H-benzotriazole (16.2 g, 0.10 mol) is dis potassium iodide (0.05 mol) were respectively added. The solved into 100 ml of 30% wt. sodium hydroxide, 3-chloro mixture was stirred for 10 min at ambient temperature, and bromopropane (31.4g, 0.10 mol), tetrabutyl ammonium bro then heated and refluxed to react for 15 hours. The mixture mide (0.8 g) are added, and mixed for 5 min. The reaction was cooled down to ambient temperature and filtered. The solution is gradually heated to 60°C., stirred for reaction for filtrate was concentrated to produce oily products, and treated 2 hours. Post treatment was performed based on common method two for synthesis. The Solution was separated and by chromatography with neutral Al-O, purified, eluted with purified by HPLC to produce 7.9 g of 1-(3-chloropropyl)-6- dichloromethane/methanol mixture to produce 12.67 g N-(2- formoxyl-1H-benzotriazole, with a yield of 33.2%. (1-benzotriazolyl)ethyl)-4-(3-(6-fluorobenzisoxazolyl))pip 25 eridine (I-143) with a yield of 69.4%. ESI-MS M+H: m/z Preparation of N-(3-(6-formoxyl-1H-benzotriazole 3652. 1-yl)propyl)-4-(3-(6-fluorobenzisoxazolyl))piperi dine (I-141) Example 120 1-(3-chloropropyl)-6-formoxyl-1H-benzotriazole (7.13 g, 30 Preparation of N-(4-(1-benzotriazolyl)butyl)-4-(3-(6- 0.03 mol) was dissolved into 150 ml of acetonitrile, 6-fluoro fluorobenzisoxazolyl))piperidine (I-144) 3-(piperidine-4-yl)benzisoxazole (5.5 g., 0.025 mol), diiso propylethylamine (12.9 g, 0.1 mol) and potassium iodide The method described in Example 101 was adopted to (4.15g, 0.025 mol) were respectively added. The mixture was prepare 1-(4-chlorobutyl)-1H-benzotriazole. stirred and mixed for 10 min at ambient temperature, then 35 1-(4-chlorobutyl)benzotriazole (0.06 mol) was dissolved heated and refluxed to react for 15 hours. After treatment into 150 ml of acetonitrile, 4-(3-(6-fluoro benzisoxazolyl)) according to common method three for synthesis produced piperidine (0.05 mol), diisopropylethylamine (0.2 mol) and 7.5 g compound (I-141), with a yield of 73.6%. ESI-MS potassium iodide (0.05 mol) were respectively added. The M+H": m/z 408.2. mixture was stirred for 10 min at ambient temperature, and 40 then heated and refluxed to react for 15 hours. The mixture Example 118 was cooled down to ambient temperature and filtered. The Preparation of N-(3-(6-methoxyl benzotriazolyl) filtrate was concentrated to produce oily products, and treated propyl)-4-(3-benzisoxazolyl)piperidine (I-142) by chromatography with neutral Al2O, purified, eluted with dichloromethane/methanol mixture to produce 13.96 g N-(4- The method described in Example 116 was adopted to 45 (1-benzotriazolyl)butyl)-4-(3-(6-fluorobenzisoxazolyl))pip prepare N-(3-chloropropyl)-6-methoxyl benzotriazole. eridine (I-144) with a yield of 71.0%. ESI-MS M+H: m/z N-(3-chloropropyl)-6-methoxyl benzotriazole (0.06 mol) 3932. was dissolved into 150 ml of acetonitrile, 4-(3-benzisox azolyl) piperidine (0.05 mol), diisopropylethylamine (0.2 Example 121 mol) and potassium iodide (0.05 mol) were respectively 50 added. The mixture was stirred for 10 min at ambient tem Preparation of N-(4-(6-cyanobenzotriazolyl)butyl)- perature, and then heated and refluxed to react for 15 hours. 4-(3-(6-fluorobenzisoxazolyl)piperidine (I-145) The mixture was cooled down to ambient temperature and filtered. The filtrate was concentrated to produce oily prod Preparation of 1-(3-chloro ucts, and treated by chromatography with neutral Al-O puri 55 butyl)-6-cyano-1H-benzotriazole fied, eluted with dichloromethane/methanol mixture to pro duce 13.14 g N-(3-(6-methoxyl benzisoxazolyl) propyl)-4- 6-cyano-1H-benzotriazole (15.9 g, 0.10 mol) is dissolved (3-benzisothiazolyl)piperidine (I-142) with a yield of 67.2%. into 100 ml of 30% wt. sodium hydroxide, 3-chlorobromobu ESI-MSM--H: m/z 391.2. tane (32.6 g., 0.10 mol), tetrabutyl ammonium bromide (0.8 g) 60 are added, and mixed for 5 min. The reaction solution is Example 119 gradually heated to 60° C., stirred for reaction for 2 hours. Post treatment was performed based on common method one Preparation of N-(2-(1-benzotriazolyl)ethyl)-4-(3-(6- for synthesis. The solution was separated and purified by fluorobenzisoxazolyl)piperidine (I-143) HPLC to produce 9.1 g of 1-(3-chlorobutyl)-6-cyano-1H 65 benzotriazole, with a yield of 32.6%. Benzotriazole (11.9 g, 0.10 mol) is dissolved into 100 ml of 1-(3-chlorobutyl)-6-cyano benzotriazole (0.06 mol) was 30% wt. sodium hydroxide, 3-chlorobromopropane (31.4g, dissolved into 150 ml of acetonitrile, 4-(3-(6-fluorobenzisox US 9,415,047 B2 99 100 azolyl)) piperidine (0.05 mol), diisopropylethylamine (0.2 was dissolved into 150 ml of acetonitrile, 4-(3-benzisox mol) and potassium iodide (0.05 mol) were respectively azolyl) piperidine (0.05 mol), diisopropylethylamine (0.2 added. The mixture was stirred for 10 min at ambient tem mol) and potassium iodide (0.05 mol) were respectively perature, and then heated and refluxed to react for 15 hours. added. The mixture was stirred for 10 min at ambient tem The mixture was cooled down to ambient temperature and perature, and then heated and refluxed to react for 15 hours. filtered. The filtrate was concentrated to produce oily prod The mixture was cooled down to ambient temperature and ucts, and treated by chromatography with neutral Al-O puri filtered. The filtrate was concentrated to produce oily prod fied, eluted with dichloromethane/methanol mixture to pro ucts, and treated by chromatography with neutral Al-O puri duce 15.07 g N-(4-(6-cyanobenzotriazolyl)butyl)-4-(3-(6- fied, eluted with dichloromethane/methanol mixture to pro fluoro benzisoxazolyl))piperidine (I-145) with a yield of 10 duce 12.88 g N-(2-(1-benzisoxazolyl) ethoxyl)-4-(3-fluoro 72.1%. ESI-MSM--H": m/z 418.2. benzisothiazolyl) piperidine (I-148) with a yield of 67.6%. ESI-MSM--H": m/z. 364.2. Example 122 Example 125 Preparation of N-(4-(6-cyanobenzotriazolyl)butyl)- 15 4-(3-(6-methoxyl benzisoxazolyl))piperidine (I-146) Preparation of N-(3-(6-methoxyl benzotriazolyl) propyl)-4-(3-(6-fluorobenzisothiazolyl))piperidine The method described in Example 121 was adopted to (I-149) prepare 1-(3-chlorobutyl)-6-cyanobenzotriazole. 1-(3-chlorobutyl)-6-cyano benzotriazole (0.06 mol) was The method described in Example 116 was adopted to dissolved into 150 ml of acetonitrile, 4-(3-(6-methoxyl ben prepare N-(3-chloropropyl)-6-methoxyl benzotriazole. Zisoxazolyl)) piperidine (0.05 mol), diisopropylethylamine N-(3-chloropropyl)-6-methoxyl benzotriazole (0.06 mol) (0.2 mol) and potassium iodide (0.05 mol) were respectively was dissolved into 150 ml of acetonitrile, 4-(3-(6-fluoroben added. The mixture was stirred for 10 min at ambient tem Zisothiazolyl)) piperidine (0.05 mol), diisopropylethylamine perature, and then heated and refluxed to react for 15 hours. 25 (0.2 mol) and potassium iodide (0.05 mol) were respectively The mixture was cooled down to ambient temperature and added. The mixture was stirred for 10 min at ambient tem filtered. The filtrate was concentrated to produce oily prod perature, and then heated and refluxed to react for 15 hours. ucts, and treated by chromatography with neutral Al-O puri The mixture was cooled down to ambient temperature and fied, eluted with dichloromethane/methanol mixture to pro filtered. The filtrate was concentrated to produce oily prod duce 15.01 g N-(4-(6-cyanobenzotriazolyl)butyl)-4-(3-(6- 30 ucts, and treated by chromatography with neutral Al-O puri methoxyl benzisoxazolyl)piperidine (I-146) with a yield of fied, eluted with dichloromethane/methanol mixture to pro 69.8%. ESI-MS IM+H: m/z 430.2. duce 13.17 g N-(3-(6-methoxyl benzotriazolyl)propyl)-4-(3- (6-fluorobenzisothiazolyl)piperidine (I-149) with a yield of Example 123 69.1%. ESI-MSM--H: m/z 426.1. 35 Preparation of N-(2-(6-methoxyl benzotriazolyl) Example 126 ethoxyl)-4-(3-benzisoxazolyl)piperidine (I-147) Preparation of N-(3-(6-methoxyl benzotriazolyl) The method described in common method four for synthe propyl)-4-(3-(6-fluorobenzopyrazol))piperidine sis was adopted to prepare N-hydroxyl-methoxyl benzotria 40 (I-150) Zole. The compound was prepared by using the methods for The method described in Example 116 was adopted to synthesis and after treatment with N-hydroxyl-6-methoxyl prepare N-(3-chloropropyl)-6-methoxyl benzotriazole. benzotriazole as the material. N-(2-chloro ethoxyl-6-meth N-(3-chloropropyl)-6-methoxyl benzotriazole (0.06 mol) oxyl benzotriazole (0.06 mol) was dissolved into 150 ml of 45 was dissolved into 150 ml of acetonitrile, 4-(3-(6-fluoroben acetonitrile, 4-(3-benzisoxazolyl) piperidine (0.05 mol), Zisothiazole)) (0.05 mol), diisopropylethylamine (0.2 mol) diisopropylethylamine (0.2 mol) and potassium iodide (0.05 and potassium iodide (0.05 mol) were respectively added. mol) were respectively added. The mixture was stirred for 10 The mixture was stirred for 10 min at ambient temperature, min at ambient temperature, and then heated and refluxed to and then heated and refluxed to react for 15 hours. The mix react for 15 hours. The mixture was cooled down to ambient 50 ture was cooled down to ambient temperature and filtered. temperature and filtered. The filtrate was concentrated to The filtrate was concentrated to produce oily products, and produce oily products, and treated by chromatography with treated by chromatography with neutral Al-O, purified, neutral Al-O, purified, eluted with dichloromethane/metha eluted with dichloromethane/methanol mixture to produce nol mixture to produce 14.21 g N-(2-(6-methoxylbenzisox 12.11 g N-(3-(6-methoxyl benzotriazolyl)propyl)-4-(3-(6- azolyl) ethoxyl)-4-(3-benzisothiazolyl) piperidine (I-147) 55 fluoro benzopyrazol)piperidine (I-150) with a yield of with a yield of 69.1%. ESI-MSM-i-HI": m/z 394.2. 66.5%. ESI-MSM--H: m/z 409.2. Example 124 Example 127

Preparation of N-(2-(1-benzotriazolyl)ethyl)-4-(3- 60 Preparation of N-(3-(6-methoxyl benzotriazolyl) fluorobenzisoxazolyl) piperidine (I-148) propyl)-4-(3-(6-fluorobenzofuranyl))piperidine (I-151) The method described in common method four for synthe sis was adopted to prepare N-hydroxyl benzotriazole. The method described in Example 116 was adopted to The compound was prepared by using the methods for 65 prepare N-(3-chloropropyl)-6-methoxyl benzotriazole. synthesis and after treatment with N-hydroxyl benzotriazole N-(3-chloropropyl)-6-methoxyl benzotriazole (0.06 mol) as the material. N-(2-chloroethoxylbenzotriazole (0.06 mol) was dissolved into 150 ml of acetonitrile, 4-(3-(6-fluoroben US 9,415,047 B2 101 102 Zisothiazole)) (0.05 mol), diisopropylethylamine (0.2 mol) products, and treated by chromatography with neutral Al2O, and potassium iodide (0.05 mol) were respectively added. eluted with dichloromethane/methanol mixture to produce The mixture was stirred for 10 min at ambient temperature, 6.1 g compound (I-154) with a yield of 60.1%. ESI-MS and then heated and refluxed to react for 15 hours. The mix M+H: m/z 336.2. ture was cooled down to ambient temperature and filtered. The filtrate was concentrated to produce oily products, and Example 131 treated by chromatography with neutral Al-O purified, eluted with dichloromethane/methanol mixture to produce Preparation of N-(4-(1H-benzotriazole-1-yl)butyl)-4- 12.40 g N-(3-(6-methoxyl benzotriazolyl)propyl)-4-(3-(6- cyclohexyl piperidine (I-155) fluoro benzofuranyl)piperidine (I-151) with a yield of 68.1%. ESI-MSM--H: m/z 409.2. The method described in Example 101 was adopted to prepare 1-(4-chlorobutyl)-1H-benzotriazole. Example 128 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, 4-(1-cyclohexyl) Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- 15 piperidine (5.1 g, 0.03 mol), diisopropylethylamine (15.5 g. 4-(2-furyl)piperidine (I-152) 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec tively added. The mixture was stirred and mixed, then heated The method described in Example 100 was adopted to and refluxed to react for 20 hours. The mixture was cooled prepare 1-(4-chlorobutyl)-1H-benzimidazole. down to ambient temperature and filtered. The filtrate was 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) concentrated to produce oily products, and treated by chro was dissolved into 100 ml of acetonitrile, 4-(2-furyl) piperi matography with neutral Al-O and purified, eluted with dine (4.6 g., 0.03 mol), diisopropylethylamine (15.5 g., 0.12 dichloromethane to produce 6.5 g compound (I-155) with a mol) and potassium iodide (5.0 g, 0.03 mol) were respectively yield of 63.7%. ESI-MS M+H": m/z 341.3. added. The mixture was stirred for 10 min at ambient tem perature, and then heated and refluxed to react for 20 hours. 25 Example 132 The mixture was cooled down to ambient temperature and filtered. The filtrate was concentrated to produce oily prod Preparation of N-(4-(1H-benzotriazole-1-yl)butyl)-4- ucts, and treated by chromatography with neutral Al2O, (1-naphthyl)piperidine (I-156) eluted with dichloromethane/methanol mixture to produce 6.0 g compound (I-152) with a yield of 61.6%. ESI-MS 30 The method described in Example 101 was adopted to M+H": m/z 324.2. prepare 1-(4-chlorobutyl)-1H-benzotriazole. 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) Example 129 was dissolved into 100 ml of acetonitrile, 4-(1-naphthyl)pip eridine (6.4g, 0.03 mol), diisopropylethylamine (15.5g, 0.12 Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- 35 mol) and potassium iodide (5.0g, 0.03 mol) were respectively 4-(4-pyridyl)piperidine (I-153) added. The mixture was stirred and mixed, then heated and refluxed to react for 20 hours. The mixture was cooled down The method described in Example 100 was adopted to to ambient temperature and filtered. The filtrate was concen prepare 1-(4-chlorobutyl)-1H-benzimidazole. trated to produce oily products, and treated by chromatogra 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) 40 phy with neutral Al-O and purified, eluted with dichlo was dissolved into 100 ml of acetonitrile, 4-(4-pyridyl) pip romethane to produce 6.9 g compound (I-156) with a yield of eridine (4.9 g, 0.03 mol), diisopropylethylamine (15.5g, 0.12 60.1%. ESI-MSM--H: m/z 385.3. mol) and potassium iodide (5.0 g, 0.03 mol) were respectively added. The mixture was stirred for 10 min at ambient tem Example 133 perature, and then heated and refluxed to react for 20 hours. 45 The mixture was cooled down to ambient temperature and Preparation of N-(4-(1H-benzotriazole-1-yl)butyl)-4- filtered. The filtrate was concentrated to produce oily prod (2-quinoxalinyl)piperidine (I-157) ucts, and treated by chromatography with neutral Al2O, eluted with dichloromethane/methanol mixture to produce The method described in Example 101 was adopted to 6.3 g compound (I-153) with a yield of 62.1%. ESI-MS 50 prepare 1-(4-chlorobutyl)-1H-benzotriazole. M+H": m/z 335.2. 1-(4-chlorobutyl)-1H-benzotriazole (7.55 g, 0.036 mol) was dissolved into 100 ml of acetonitrile, 4-(2-quinoxalinyl) Example 130 piperidine (6.4g, 0.03 mol), diisopropylethylamine (15.5 g. 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec Preparation of N-(4-(1H-benzimidazoyl-1-yl)butyl)- 55 tively added. The mixture was stirred and mixed, then heated 4-(2-pyrimidinyl)piperidine (I-154) and refluxed to react for 20 hours. The mixture was cooled down to ambient temperature and filtered. The filtrate was The method described in Example 100 was adopted to concentrated to produce oily products, and treated by chro prepare 1-(4-chlorobutyl)-1H-benzimidazole. matography with neutral Al-O and purified, eluted with 1-(4-chlorobutyl)-1H-benzimidazole (7.51 g, 0.036 mol) 60 dichloromethane to produce 7.3 g compound (I-157) with a was dissolved into 100 ml of acetonitrile, 4-(2-pyrimidinyl) yield of 62.7%. ESI-MS M+H: m/z 387.2. piperidine (4.9 g, 0.03 mol), diisopropylethylamine (15.5 g. 0.12 mol) and potassium iodide (5.0g, 0.03 mol) were respec Example 134 tively added. The mixture was stirred for 10 min at ambient temperature, and then heated and refluxed to react for 20 65 Relaxing effects of compound II-1 to II-60 and II-84 to hours. The mixture was cooled down to ambient temperature II-157 on vascular smooth muscle constricted by convulsant and filtered. The filtrate was concentrated to produce oily in vitro US 9,415,047 B2 103 104 1. Experimental Animals: Surrounding fatty tissues (for hydroxytryptamine receptor Rabbits, male or female, 2.0-3.0 kg, were provided by antagonism test, endothelia should be removed with Smooth Experimental Animal Center, China Medical university. stainless steel rod) removed, the aorta was cut into 3-5 mm 2. Drugs and Reagents vascular rings which were penetrated together with a steel Compound II-1 to II-60 and II-84 to II-157, i.e., salts (hy wire with one end fixed on ventilation hook, another end on drochlorates) prepared by using the above methods, were pressure tonotransducer. The aortarings were put into a bath used for the following experiments. tube with 20 ml of nutritional solution, and the tension Sodium chloride (NaCl): bought from Tianjin Damao changes were recorded by using the recorder. The tempera Chemical Reagent Factory, batch number 20120413. ture of bathtube was maintained at 37+0.5°C., and mixed gas Kalium chloride (NaCl): bought from Tianjin Damao 10 Chemical Reagent Factory, batch number 2011 1123. (95% O+5% CO) was ventilated at a rate of I-2 bubbles/ Anhydrous magnesium Sulfate (MgSO4): bought from second. The initial load of the sample was 1.5 g, nutritional Tianjin Damao Chemical Reagent Factory, batch number Solution was changed for every 20 min. The sample was 20101029. balanced for 2 hours, and the experiment would be started when baseline became stable. Anhydrous calcium chloride (CaCl): bought from Tianjin 15 Damao Chemical Reagent Factory, batch number 2011 0314. 6. Specific Experimental Procedure and Results Sodium bicarbonate (NaHCO): bought from Tianjin 6.1 Relaxing Effects of Compound II-1 to II-60 and II-84 to Damao Chemical Reagent Factory, batch number 20120507. Glucose: bought from Tianjin Damao Chemical Reagent II-157 on Vascular Smooth Muscle Constricted by Convul Factory, batch number 20120512. sant Adrenaline Hydrochloride (AD) In Vitro Kalium dihydrogen phosphate (KHPO): bought from After sample tension became stable, a piece of waveform Tianjin Damao Chemical Reagent Factory, batch number was recorded. Adrenaline hydrochloride (AD) (10 mol/L) 20110928. was added into a bath tube to induce constriction, when Sodium chloride injection (NaCl): bought from Shenyang maximal constriction was achieved, the sample was com Zhiying Pharmaceutical Factory, batch number: 12021001. 25 pletely flushed, K-H solution was changed for every 20 min. Epinephrine hydrochloride injection: strength: 1 mg/1 ml, Contents in the tube was balanced for 60 min, when baseline bought from Grandpharma (China) Co., Ltd., batch number: recovered to be stable, the convulsant was added at the same 12O105. concentration to induce constriction. When the later maximal Norepinephrine bitartrate injection: strength: 2 mg/1 ml, constriction response was basically consistent with the bought from Grandpharma (China) Co., Ltd., batch number: 30 former one, the prepared compound solutions were Subse 12O3O4. quently added, including compound II-1 to II-60 solutions 3. Experimental Instruments HSS-1 (B) thermostat bath: Chengdu Instrument Factory. (1x10-1x10 mol/L), compound II-84 to II-123 solutions RM6240B multi-channel physiological signal collection (1x10'-1x10° mol/L) and compound II-124 to II-157 & processing system: Chengdu Instrument Factory. 35 solutions (1x10'-1x10 mol/L), to record the waveforms. JZJO1 muscular strength transducer: Chengdu Instrument Relaxing percentage of the compound was used as Y axis, Factory. maximal relaxation response was 100%, negative logarithm YPJO1 pressure transducer: Chengdu Instrument Factory. of different concentration was used as X axis, to draw dose TG-328A photoelectric analytical balance: Shanghai Bal effect curve (expressed in meant-SEM (--), n=5). Com ance Factory. 40 pounds II-2 and II-3 had the most obvious relaxing effects T-500 electronic balance: Chuangshu Shuangjie Test (FIG. 1 and FIG. 8). Instrument Factory. From FIG. 1 and FIG. 8, compounds II-2 and II-3 had Micropipette: Shanghai Rongtai Biochemical Engineering relaxing effects on samples constricted by AD in certain dose Co., Ltd. dependent manner. The -log ECso value was 5.73+0.03 for electrical heated thermostatic water bath: Tianjin Taisite 45 compound II-2 to relax adrenaline constricted rabbit aorta, Instrument Co., Ltd. and 6.01 +0.05 for compound II-3. 4. Preparation of Nutrient Solution Krebs-Henseleit (K-H) normal saline: NaCl 6.92 (concen Compound II-31 also had relatively obvious relaxing tration unit), KCl 0.35, MgSO, 0.29, KHPO, 0.16, CaCl, effects, negative logarithms of different concentrations were 0.28, NaHCO, 2.1, Glucose 2.0 (g/L), pH 7.2. 50 used as X axis to draw dose effective curves, which were High kalium solution: removing equal mole of NaCl from shown in FIG. 10. II-29 also had obvious relaxing effects, and K-H solution, adding KCl to prepare modified K-H solution its dose effect curve was shown in FIG. 15. From FIG. 10, containing K"60 mmol/L. compound II-31 had relaxing effects on samples constricted K-H free solution: removing CaCl from K-H solution, by AD in certain dose dependent manner. The -log ECso adding equal mole of KCl and EDTA'Na'0.1 mmol/L, other 55 value was 6.19+0.03 for compound II-31 to relax AD con components not changed. stricted rabbit aorta. Similarly, the relaxing effects of com Calcium free high kalium solution: removing CaCl2 from pound II-29 against AD was also in a dose dependent manner. high kalium solution, adding equal mole of KCl and EDTA The -log ECso value was 6.01+0.02 for compound II-29 to Na"0.1 mmol/L, other components not changed. relax AD constricted rabbit aorta. Preparation of compound II-1 to II-60 and II-84 to II-157 60 Solutions: weigh appropriate amount of compound sample, From FIG. 17, compound II-85 had relaxing effects on dilute to solutions of series concentrations with distilled vascular samples constricted by AD in certain dose dependent water (109-10 mol/L) for later use. manner. The -log ECso value was 7.30+0.05 for compound 5. Preparation of Excised Vascular Smooth Muscle Sample II-85 to relax AD constricted rabbit aorta. from Rabbits 65 Relaxing effects of compound II-1 to II-60, II-84 to II-123 Rabbits were hit to be dizzy, with thoracic cavity quickly and II-124 to II-157 on vascular smooth muscle constricted cut open, descending aorta exposed, connective tissues and by AD were shown in table 1: US 9,415,047 B2 105 106 TABLE 1. TABLE 1-continued Compound -logECso Compound -logECso 5.03 - 0.04 -102 4.26 - 0.06 5.73 O.O3 -103 5.21 O.OS 6.01 - 0.05 -104 5.02 O.04 4.96 - 0.03 -105 5.18 O.O3 4.78 O.04 -106 5.21 O.OS 4.63 - 0.06 -107 5.03 O.O2 4.29 O.OS -108 5.16 O.O3 4.71 - 0.04 10 -109 6.21 - 0.04 4.37 - O.O3 -110 6.36 0.03 4.26 - 0.05 -111 489 - O.O2 4.05 - 0.04 -112 4.76 O.O3 4.35 - 0.06 -113 5.31 O.04 4.41 - 0.05 -114 4.86 - 0.03 4.22 O.04 15 -115 479 0.02 4.47 0.04 -116 S.S6 O.OS 4.29 O.O3 -117 5.31 OO6 4.53 - 0.03 -118 4.43 O.OS 4.86 - 0.06 -119 4.86 - 0.04 4.18 O.04 -12O 4.72, O.O3 4.23 - 0.05 -121 4.39 O.OS 4.05 - 0.03 -122 4.22 OO6 4.550.04 -123 4.83 - 0.05 4.72, O.O3 -124 5.03 O.O2 4.52 0.04 -12S 5.16 O.O3 4.79 0.05 -126 6.21 - 0.04 419 0.04 -127 6.36 0.03 4.31 - 0.04 25 -128 489 - O.O2 3.99, O.O3 -129 4.76 O.O3 6.01 - O.O2 -130 5.31 O.04 S.S2 O.O3 -131 4.86 - 0.03 6.19 O.O3 -132 479 0.02 541 O.O3 -133 S.S6 O.OS 4.39 O.04 30 -134 5.31 OO6 8.07 - 0.06 -13S 5.45 O.04 4.89 0.05 -136 5.34 O.O3 5.31 O.04 -137 5.61 O.OS S.S6 O.O3 -138 5.42 O.04 5.72 O.OS -139 5.38 O.O3 547 O.04 35 -140 S.23 O.OS 5.35 0.06 -141 5.56 O.04 4.51 - 0.05 -142 6.11 - O.O7 4.39 O.04 -143 5.92 O.OS 4.45 0.04 -144 5.96 O.04 415 0.03 -145 S.S3 O.O7 4.33 - 0.03 -146 5.23 OO6 4.26 - 0.06 40 -147 4.03 - 0.05 3.88 O.04 -148 4.26 - 0.04 3.83 O.OS -149 4.01 - 0.03 4.05 - 0.03 -1SO 4.13 - 0.05 4.35 - 0.04 -151 4.26 - 0.06 4.52 0.03 -152 4.43 O.OS 4.88 0.04 45 -153 4.86 - 0.04 4.28 0.05 -154 4.72, O.O3 5.21 O.04 -155 4.39 O.OS 4.01 - 0.03 -156 4.22 OO6 4.26 - 0.05 -157 4.83 - 0.05 4.21 O.O3 4.17 O.04 50 4.53 - 0.05 6.2 Relaxing Effects of Compound II-84 to II-157 on Vas 4.05 - 0.04 6.23 - 0.04 cular Smooth Muscle Constricted by Noradrenaline (NA) In 7.30 O.OS Vitro 5.45 O.04 After sample tension became stable, a piece of waveform 5.34 O.O3 55 5.61 O.OS was recorded. Adrenaline hydrochloride (AD) (10 mol/L) 542, O.O4 was added into a bath tube to induce constriction, when 5.38 O.O3 maximal constriction was achieved, the sample was com S.23 O.OS pletely flushed, K-H solution was changed for every 20 min. 5.56 O.04 6.11 - O.O7 Contents in the tube was balanced for 60 min, when baseline 5.92 O.OS 60 recovered to be stable, noradrenaline bitartrate (NA) (10 5.96 O.04 mol/L) was added to induce constriction. When the later S.S3 O.O7 maximal constriction response was basically consistent with 5.23 OO6 4.03 - 0.05 the former one, the prepared compound solutions were Sub 4.26 - 0.04 sequently added, including compound II-84 to II-123 solu 4.01 - 0.03 65 tions (1x10'-1x10° mol/L), compound II-124 to II-157 4.13 - 0.05 solutions (1x10'-1x10 mol/L), to record the waveforms. Relaxing percentage of the compound was used as Y axis, US 9,415,047 B2 107 108 maximal relaxation response was 100%, negative logarithm TABLE 2-continued of different concentration was used as X axis, to draw dose effect curve (expressed in meant-SEM (--), n=5). Compound -logECso From FIG. 18, compound II-85 had relaxing effects on I-148 4.04 O.O7 vascular samples constricted by NA in certain dose dependent 5 I-149 4.19 O.O3 I-1SO 4.43 - 0.04 manner. The -log ECso value was 7.51+0.05 for compound I-151 4.06 - 0.06 II-85 to relax NA constricted rabbit aorta. I-152 4.23 O.O3 Relaxing effects of compound II-84 to II-157 on vascular I-153 4.66 O.O2 smooth muscle constricted by NA in vitro were shown in table I-154 4.52 0.04 2: 10 I-155 4.44 - 0.05 I-156 4.34 - 0.04 I-157 4.53 - 0.05 TABLE 2 Compound -logECso 6.3 Relaxing Effects of Compound II-1 to II-60 and II-84 to 15 -84 6.17 O.O3 II-157 on Vascular Smooth Muscle Constricted by High -85 7.51 O.OS Kalium Concentration. In Vitro -86 5.26 O.04 -87 5.04 O.O3 After sample tension became stable, a piece of waveform -88 5.73 O.OS was recorded. Adrenaline hydrochloride (AD) (10 mol/L) -89 5.22 O.04 -90 5.35 O.O3 2O was added into a bath tube to induce constriction, when -91 5.31 O.OS maximal constriction was achieved, the sample was com -92 5.73 O.04 pletely flushed, K-H solution was changed for every 20 min. -93 6.07 - 0.04 -94 5.81 - O.O3 Contents in the tube was balanced for 60 min, when baseline -95 5.73 O.04 recovered to be stable, K-H solution in the bath tube was -96 5.31 OO6 25 replaced with high kalium concentration solution to induce -97 5.09 OO4 -98 4.31 - 0.05 constriction. When the later maximal constriction response -99 4.04 O.O7 was basically consistent with the former one, the prepared -100 4.19 O.O3 compound solutions were Subsequently added, including -101 4.43 - 0.04 compound II-1 to II-60 solutions (1x10-1x10 mol/L), -102 4.06 - 0.06 30 -103 5.11 - O.OS compound II-84 to II-123 solutions (1x10'-1x10 mol/L) -104 5.02 O.04 and compound II-124 to II-157 solutions (1x10'-1x10 -105 5.11 - O.O3 mol/L), to record the waveforms. Relaxing percentage of the -106 5.10 O.OS -107 5.11 - O.O2 compound was used as Y axis, maximal relaxation response -108 S.27 O.O3 35 was 100%, negative logarithm of different concentration was -109 6.32 0.04 used as X axis, to draw dose effect curve (expressed in -110 6.45 0.03 -111 4.67 + O.O2 meant-SEM (--), n=5). Compounds II-2 and II-3 had the -112 4.55 0.03 most obvious relaxing effects (FIG. 2 and FIG. 9). -113 5.21 O.04 -114 4.77 0.03 From FIG. 2 and FIG. 9, compounds II-2 and II-3 had -115 4.53 O.O2 40 obvious relaxing effects on Samples constricted by high -116 5.36 O.OS kalium concentration Solution in certain dose dependent man -117 5.15 OO6 -118 4.23 O.O3 ner. The -log ECso value was 5.34+0.02 for compound II-2 to -119 4.66 O.O2 relax high kalium concentration constricted rabbit aorta, and -12O 4.52 0.04 5.49+0.05 for compound II-3. Of these compounds, com -121 4.44 - 0.05 45 pound II-31 also had relatively obvious relaxing effects, -122 4.34 - 0.04 -123 4.53 - 0.05 negative logarithms of different concentrations were used as -124 5.11 - O.O2 X axis to draw dose effective curves, which were shown in -12S S.27 O.O3 FIG. 11. II-29 also had obvious relaxing effects, and its dose -126 6.32 0.04 -127 6.45 0.03 50 effect curve was shown in FIG. 16. From FIG. 11, compound -128 4.67 + O.O2 II-31 had relaxing effects on samples constricted by high -129 4.55 0.03 kalium concentration in certain dose dependent manner. The -130 5.21 O.04 -131 4.77 0.03 -log ECs value was 5.55+0.03 for compound II-31 to relax -132 4.53 O.O2 high kalium concentration constricted rabbit aorta. Similarly, -133 5.36 O.OS 55 the relaxing effects of compound II-29 against high kalium -134 5.15 OO6 -13S 5.26 O.04 concentration was also in a dose dependent manner. The -log -136 5.04 O.O3 ECso value was 5.64+0.01 for compound II-29 to relax high -137 5.73 O.OS kalium concentration constricted rabbit aorta. -138 5.22 O.04 -139 5.35 O.O3 From FIG. 19, compound II-85 had relaxing effects on -140 5.31 O.OS 60 vascular samples constricted by high kalium concentration in -141 5.73 O.04 -142 6.07 - 0.04 certain dose dependent manner. The -log ECso value was -143 5.81 - O.O3 6.21+0.03 for compound II-85 to relax high kalium concen -144 5.73 O.04 tration constricted rabbit aorta. -145 5.31 OO6 -146 5.09 OO4 65 Relaxing effects of compound II-1 to II-60 and II-84 to -147 4.31 - 0.05 II-157 on vascular smooth muscle constricted by high kalium concentration in vitro were shown in table 3: US 9,415,047 B2 109 110 TABLE 3 TABLE 3-continued Compound -logECso Compound -logECso S.O.S. O.O3 -102 3.46 O.O3 5.34 O.O2 5 -103 3.21 O.OS 549 O.OS -104 3.42 O.04 4.79 0.05 -105 3.38 O.O3 4.53 - 0.03 -106 3.23 O.OS 4.41+0.04 -107 3.69 O.O2 3.79 O.O3 -108 3.82 O.O3 4.41 - 0.05 10 -109 5.01 - O.04 4.28 0.03 -110 5.12, O.O3 3.96 O.OS -111 3.44 O.O2 3.85 O.04 -112 3.38 O.O3 415 0.06 -113 4.03 - 0.04 4.52 0.05 -114 3.56 O.O3 4.05 - 0.04 15 -115 3.23 O.O2 4.52 0.05 -116 4.22 O.OS 4.19 O.O3 -117 4.17 O.O6 4.31 - 0.04 -118 3.53 O.O3 4.74 - 0.06 -119 3.26 O.04 4.06 - 0.03 -12O 3.32. O.O2 3.93 + O.O2 -121 3.14 O.OS 3.75 O.O3 2O -122 3.04 O.O3 4.64 - 0.04 -123 3.13 - 0.04 4.42 0.05 -124 4.69 O.O2 4.52 0.04 -12S 4.82 0.03 4.53 - 0.03 -126 6.01 - 0.04 3.99, O.OS -127 6.12 O.O3 4.06 - 0.04 25 -128 4.44 + 0.02 3.85 O.04 -129 4.38 0.03 5.64 OO1 -130 5.03 - 0.04 5.13 - O.O3 -131 4.56 0.03 S.S.S. O.O3 -132 4.23 O.O2 461 O.O3 -133 5.22 O.OS 3.94 O.04 30 -134 5.17 OO6 4.77 O.O2 -13S 5.09 OO4 449 0.05 -136 5.12, O.O3 5.31 O.04 -137 5.72 O.OS 5.43 O.O3 -138 5.12 O.04 5.33 O.04 -139 5.28, O.O3 5.22 O.04 35 -140 S.O2 O.OS 5.29 O.04 -141 5.32 O.04 461 - 0.05 -142 5.11 - O.O3 3.93 + O.04 -143 3.92 O.O2 3.85 O.04 -144 3.96 O.O3 3.73 O.O3 -145 3.53 O.O2 4.09, O.O3 -146 4.23 - 0.04 3.92 O.O2 40 -147 3.53 O.O3 3.54 O.O3 -148 4.26 - 0.04 3.43 O.04 -149 3.31 O.O3 3.85 O.O3 -1SO 3.63 O.04 3.79 0.04 -151 3.46 O.O3 4.46 - 0.03 -152 4.53 - 0.03 4.58 0.04 45 -153 4.26 - 0.04 3.88 O.O2 -154 4.32 O.O2 4.91 - 0.04 -155 4.14 - 0.05 3.71 - O.O3 -156 4.04 O.O3 3.51 O.O2 -157 4.13 - 0.04 3.58 O.O2 3.75 O.04 50 4.21 O.O3 3.81 - O.O2 Example 135 4.27- 0.04 5.21 O.O3 4.09 0.04 Study on Mechanism of Relaxing Effects of 4.12 O.O3 55 Compound II-2 and II-85 on Vascular Smooth 4.72 0.05 Muscle In Vitro 4.12 0.04 4.28 0.03 4.02 0.05 1. Experimental Animals: 4.32 0.04 4.11 - 0.03 Rabbits, male or female, 2.0-3.0 kg, were provided by 3.92 O.O2 60 Experimental Animal Center, China Medical university. 3.96 O.O3 2. Drugs and Reagents 3.53 O.O2 Methods described in Example 2 and 72 were adopted to 3.23 O.04 3.53 O.O3 prepare compound II-2 and II-85. 3.26 O.04 Sodium chloride (NaCl): bought from Tianjin Damao 3.31 O.O3 65 Chemical Reagent Factory, batch number 20120413. 3.63 O.04 Kalium chloride (NaCl): bought from Tianjin Damao Chemical Reagent Factory, batch number 2011 1123. US 9,415,047 B2 111 112 Anhydrous magnesium Sulfate (MgSO4): bought from dilute to solutions of series concentrations with distilled Tianjin Damao Chemical Reagent Factory, batch number water (109-10 mol/L) for later use. 20101029. 5. Preparation of Excised Vascular Smooth Muscle Sample Anhydrous calcium chloride (CaCl): bought from Tianjin from Rabbits Damao Chemical Reagent Factory, batch number 2011 0314. Rabbits were hit to be dizzy, with thoracic cavity quickly Sodium bicarbonate (NaHCO): bought from Tianjin cut open, descending aorta exposed, connective tissues and Damao Chemical Reagent Factory, batch number 20120507. Surrounding fatty tissues (for hydroxytryptamine receptor Glucose: bought from Tianjin Damao Chemical Reagent antagonism test, endothelia should be removed with Smooth Factory, batch number 20120512. stainless steel rod) removed, the aorta was cut into 3-5 mm Kalium dihydrogen phosphate (KHPO): bought from 10 vascular rings which were penetrated together with a steel Tianjin Damao Chemical Reagent Factory, batch number wire with one end fixed on ventilation hook, another end on 20110928. pressure tonotransducer. The aortarings were put into a bath Sodium chloride injection (NaCl): bought from Shenyang tube with 20 ml of nutritional solution, and the tension Zhiying Pharmaceutical Factory, batch number: 12021001. changes were recorded by using the recorder. The tempera Epinephrine hydrochloride injection: strength: 1 mg/1 ml, 15 ture of bathtube was maintained at 37+0.5°C., and mixed gas bought from Grandpharma (China) Co., Ltd., batch number: (95% O+5% CO) was ventilated at a rate of I-2 bubbles/ 12O105. second. The initial load of the sample was 1.5 g, nutritional Norepinephrine bitartrate injection: strength: 2 mg/1 ml, Solution was changed for every 20 min. The sample was bought from Grandpharma (China) Co., Ltd., batch number: balanced for 2 hours, and the experiment would be started 12O3O4. when baseline became stable. Doxazosin mesylate: bought from Suizhou hake Pharma 6. Experimental Procedure and Results ceutical and Chemical Industry Co., Ltd., batch number: 6.1 Antagonism of Compound II-2 and II-85 on C.-Recep 2011 O305. tor Agonist of Vascular Smooth Muscle of Rabbits Amlodipine besylate tablets: bought from Pfizer, strength: 6.1.1 Effects of Compound II-2 on Dose Effective Curve of 5 mg/tablet, batch number: 1205018. Epinephrine hydrochlo 25 Accumulative Constriction by Noradrenaline ride injection: strength: 1 mg/1 ml, bought from Grandp After sample tension became stable, a piece of waveform harma (China) Co., Ltd., batch number: 120105. was recorded, noradrenaline (NA) (10-10" mol/L) was (R)-phenylephrine hydrochloride, bought from Tokyo added into the bath tube until maximal response, then wave Chemical Industry (Shanghai), batch number: GJ01-TESP. form was recorded. Then K-H solution was used to flush the Serotonin creatinine sulfate monohydrate (5-HT), bought 30 sample repeatedly, after balanced for 1 hour, compound II-2 from Tokyo Chemical Industry, batch number: AZ01-TBKD. (3x107 mol/L) was added. NA was also added by using the Heparin sodium injection: bought from China Wanbang, same method 20 min later. The maximal response was con strength: 2 ml/12500 unit, batch number: 101115. sidered 100%, NA constriction percentage was used as Yaxis, Urethane: Shanghai Chemical Reagent Company, China negative logarithms of different concentrations were used as National Pharmaceutical Group, batch number: C30191228. 35 X axis to draw dose effective curve. The curve was shown in EDTA: bought from Tianjin Damao Chemical Reagent FIG.3 after compound II-2 (3x107 mol/L) was added. Factory, batch number 20050809. NA dose effective curve was obviously moved to right in 3. Experimental Instruments parallel, with maximal response nearly unchanged. Percent HSS-1 (B) thermostat bath: Chengdu Instrument Factory. ages at different concentrations were statistically analyzed by RM6240B multi-channel physiological signal collection 40 using t test, most P values <0.01, Suggesting significant dif & processing system: Chengdu Instrument Factory. ference. The PA value was 7.37+0.08 for compound II-2 to JZJO1 muscular strength transducer: Chengdu Instrument resist constriction of rabbit aorta by NA. Factory. 6.1.2 Effects of Positive Reference Drug Doxazosin on YPJO1 pressure transducer: Chengdu Instrument Factory. Dose Effective Curve of Accumulative Constriction by Nora TG-328A photoelectric analytical balance: Shanghai Bal 45 drenaline ance Factory. After sample tension became stable, a piece of waveform T-500 electronic balance: Chuangshu Shuangjie Test was recorded, noradrenaline (NA) (10-10 mol/L) (10 Instrument Factory. 3x10 mol/L) was added into the bath tube until maximal Micropipette: Shanghai Rongtai Biochemical Engineering response, then waveform was recorded. Then samples were Co., Ltd. 50 flushed with K-H solution repeatedly, K-H solution was electrical heated thermostatic water bath: Tianjin Taisite changed for every 20 min, and the samples were balanced for Instrument Co., Ltd. 60 min. After baseline recovered to be stable, doxazosin (107 4. Preparation of Nutrient Solution mol/L) was added, and noradrenaline (NA) (10-6x10 Krebs-Henseleit (K-H) normal saline: NaCl 6.92 (concen mol/L) was added by using the same method 15 min later. The tration unit), KCl 0.35, MgSO, 0.29, KHPO, 0.16, CaCl, 55 maximal response was considered 100%, NA constriction 0.28, NaHCO, 2.1, Glucose 2.0 (g/L), pH 7.2. percentage was used as Y axis, negative logarithms of differ High kalium solution: removing equal mole of NaCl from ent concentrations were used as X axis to draw dose effective K-H solution, adding KCl to prepare modified K-H solution curve. The curve was shown in FIG. 4 after doxazosin (107 containing K"60 mmol/L. mol/L) was added. Dose effective curve of NA was obviously K-H free solution: removing CaCl from K-H solution, 60 moved to right in parallel, with maximal response nearly adding equal mole of KCl and EDTA'Na'0.1 mmol/L, other unchanged. Percentages at different concentrations were sta components not changed. tistically analyzed, most P values <0.01, Suggesting signifi Calcium free high kalium solution: removing CaCl from cant difference. The PA value was 7.52+0.04 for positive high kalium Solution, adding equal mole of KCl and drug doxazosin to resist constriction of rabbit aorta by NA. EDTA'Na'0.1 mmol/L, other components not changed. 65 Statistical t test showed that, for PA values of compound Preparation of compound II-2 and II-85 solutions: weigh II-2 and positive drug doxazosin against NA, P-0.05, Sug appropriate amount of compound II-2 and II-85 samples, gesting no significant difference between them, which meant US 9,415,047 B2 113 114 that compound II-2 and doxazosin had similar resisting solution for 40 min. Calcium free high kalium solution was effects against a receptor agonist. added into the sample for depolarization for 20 min, then 6.1.3 Effects of Compound II-85 on Dose Effective Curve compound II-2 (3x10" mol/L) was added into bath tube, of Accumulative Constriction by Phenephrine which was incubated for 20 min, then CaCl (10-10° mol/ After sample tension became stable, a piece of waveform 5 L) was added by using the same method till maximal response was recorded, (R)-phenylephrine hydrochloride (PE) (10 was achieved, and then waveform was recorded. The maxi 6x10 mol/L) was added into the bath tube until maximal mal response was considered 100%, CaCl constriction per response, then waveform was recorded. Then K-H solution centage was used as Y axis, negative logarithms of different was used to flush the sample repeatedly, after balanced for 1 concentrations were used as X axis to draw dose effective hour, compound II-85 (10 mol/L) was added. PE was also 10 curve. The curve was shown in FIG. 5 after compound II-2 added by using the same method 20 min later. The maximal (3x10" mol/L) was added. Dose effective curve of CaCl, was response was considered 100%, PE constriction percentage obviously moved to right in parallel, with maximal response was used as Y axis, negative logarithms of different concen nearly unchanged. Percentages at different concentrations trations were used as X axis to draw dose effective curve (data were statistically analyzed, most P values <0.01, Suggesting were expressed in meant-SEM (--), n=5). The curve was 15 significant difference. The PA value was 5.61+0.04 for com shown in FIG. 20 after compound II-85 (10 mol/L) was pound II-2 to resist constriction of rabbit aorta by CaCl. added. 6.2.2 Effects of Positive Reference Drug Amlodipine on From FIG. 20, PE dose effective curve was obviously Dose Effective Curve of Accumulative Constriction by CaCl moved to right in parallel, with maximal response nearly After sample tension became stable, calcium K-H solution unchanged. Percentages at different concentrations were sta was used to flush the sample for 3 times, and incubated with tistically analyzed by using t test, most P values <0.01, Sug calcium free K-H solution for 40 min. Calcium free high gesting significant difference. The PA value was 8.62+0.11 kalium solution was added into the sample for depolarization for compound II-85 to resist constriction of rabbit aorta by for 20 min, then CaCl, (10-10 mol/L) was added into bath PE. tube till maximal response was achieved, and then waveform 6.1.4 Effects of Positive Reference Drug Doxazosine 25 was recorded. Then K-H solution was used to flush the sample Mesylate on Dose Effective Curve of Accumulative Constric repeatedly, the K-H solution was changed for every 20 min, tion by Phenylephrine and the sample was balanced for 60 min. After sample tension After sample tension became stable, a piece of waveform became stable, calcium K-H solution was used to flush the was recorded, (R)-phenylephrine hydrochloride (PE) (10 sample for 3 times, and incubated with calcium free K-H 3x10 mol/L) was added into the bath tube until maximal 30 solution for 40 min. Calcium free high kalium solution was response, then waveform was recorded. Then K-H solution added into the sample for depolarization for 20 min, then was used to flush the sample repeatedly, after balanced for 1 amlodipine (107 mol/L) was added into bathtube, which was hour, doxazosine mesylate (10 mol/L) was added. PE was incubated for 15 min, then CaCl (10-10 mol/L) was also added by using the same method 15 min later. The added by using the same method till maximal response was maximal response was considered 100%, PE constriction 35 achieved, and then waveform was recorded. The maximal percentage was used as Y axis, negative logarithms of differ response was considered 100%, CaCl constriction percent ent concentrations were used as X axis to draw dose effective age was used as Y axis, negative logarithms of different con curve (data were expressed in meant-SEM (--), *P-0.05, centrations were used as X axis to draw dose effective curve. **P<0.01, n=7). The curve was shown in FIG. 21 after posi The curve was shown in FIG. 6 after amlodipine (3x10 tive drug doxazosine mesylate (107 mol/L) was added. 40 mol/L) was added. Dose effective curve of CaCl was obvi From FIG. 21, PE dose effective curve was obviously ously moved to right in parallel, with maximal response moved to right in parallel, with maximal response nearly nearly unchanged. Percentages at different concentrations unchanged. Percentages at different concentrations were sta were statistically analyzed, most P values <0.01, Suggesting tistically analyzed by using t test, most P values <0.01, Sug significant difference. The PA value was 6.99+0.05 for amlo gesting significant difference. The PA value was 7.43+0.12 45 dipine to resist constriction of rabbit aorta by CaCl. for positive drug doxazosin to resist constriction of rabbit 6.2.3 Effects of Compound II-85 on Dose Effect Curve of aorta by PE. CaCl on Accumulative Constriction of Rabbit Blood Vessels Statistical t test showed that, for PA values of compound After sample tension became stable, calcium K-H solution II-85 and positive drug doxazosin against PE, PK0.01, Sug was used to flush the sample for 3 times, and incubated with gesting very significant difference between them, which 50 calcium free K-H solution for 40 min. Calcium free high meant that compound II-85 had stronger resisting effects kalium solution was added into the sample for depolarization against a receptor agonist than doxazosin. for 20 min, then CaCl (10-10° mol/L) was added into bath 6.2 Antagonism of Compound II-2 and II-85 on Calcium tube till maximal response was achieved, and then waveform Channel (Ca") of Vascular Smooth Muscle of Rabbits was recorded. Then K-H solution was used to flush the sample 6.2.1 Effects of Compound II-2 on Dose Effect Curve of 55 repeatedly, the K-H solution was changed for every 20 min, CaCl on Accumulative Constriction of Rabbit Blood Vessels and the sample was balanced for 60 min. After sample tension After sample tension became stable, calcium K-H solution became stable, calcium K-H solution was used to flush the was used to flush the sample for 3 times, and incubated with sample for 3 times, and incubated with calcium free K-H calcium free K-H solution for 40 min. Calcium free high solution for 40 min. Calcium free high kalium solution was kalium solution was added into the sample for depolarization 60 added into the sample for depolarization for 20 min, then for 20 min, then CaCl, (10-10 mol/L) was added into bath compound II-85 (10 mol/L) was added into bath tube, tube till maximal response was achieved, and then waveform which was incubated for 20 min, then CaCl (10-10 mol/ was recorded. Then K-H solution was used to flush the sample L) was added by using the same method till maximal response repeatedly, the K-H solution was changed for every 20 min, was achieved, and then waveform was recorded. The maxi and the sample was balanced for 60 min. After sample tension 65 mal response was considered 100%, CaCl constriction per became stable, calcium K-H solution was used to flush the centage was used as Y axis, negative logarithms of different sample for 3 times, and incubated with calcium free K-H concentrations were used as X axis to draw dose effective US 9,415,047 B2 115 116 curve (data were expressed in meant-SEM (--), *P-0.05, bath tube until maximal response, then waveform was **P<0.01, n=7). The results were shown in FIG. 22 after recorded. Then K-H solution was used to flush the sample compound II-85 (10 mol/L) was added. repeatedly, after balanced for 1.5 hour, compound II-85 (107 From FIG.22, dose effective curve of CaCl was obviously mol/L) was added. 5-HT was also added by using the same moved to right in parallel, with maximal response nearly method 20 min later. The maximal response was considered unchanged. Percentages at different concentrations were sta 100%, 5-HT constriction percentage was used as Y axis, tistically analyzed, most P values <0.01, Suggesting signifi negative logarithms of different concentrations of 5-HT were cant difference. The PA value was 6.10+0.13 for compound used as X axis to draw dose effective curve (data were II-85 to resist constriction of rabbit aorta by CaCl. expressed in mean+SEM (--), *P-0.05, **P<0.01, n=7). 6.2.4 Effects of Amlodipine on Dose Effect Curve of CaCl 10 The curve was shown in FIG. 8 after compound II-85 (107 on Accumulative Constriction of Rabbit Blood Vessels mol/L) was added. After sample tension became stable, calcium K-H solution From FIG. 24, dose effective curve of 5-HT was obviously was used to flush the sample for 3 times, and incubated with moved to right in parallel, with maximal response nearly calcium free K-H solution for 40 min. Calcium free high unchanged. Percentages at different concentrations were sta kalium solution was added into the sample for depolarization 15 tistically analyzed, most P values <0.01, Suggesting signifi for 20 min, then CaCl, (10-10 mol/L) was added into bath cant difference. The PA value was 9.06+0.07 for compound tube till maximal response was achieved, and then waveform II-85 to resist constriction of rabbit aorta by 5-HT. was recorded. Then K-H solution was used to flush the sample repeatedly, the K-H solution was changed for every 20 min, Example 136 and the sample was balanced for 60 min. After sample tension became stable, calcium K-H solution was used to flush the Study on Mechanism of Relaxing Effects of sample for 3 times, and incubated with calcium free K-H Compound II-31 on Vascular Smooth Muscle In solution for 40 min. Calcium free high kalium solution was Vitro 25 added into the sample for depolarization for 20 min, then 1.1 Antagonism of Compound II-31 on C.-Receptor Ago amlodipine (107 mol/L) was added into bathtube, which was nist of Vascular Smooth Muscle of Rabbits incubated for 15 min, then CaCl (10-10° mol/L) was 1.1.1 Effects of Compound II-31 on Dose Effective Curve added by using the same method till maximal response was of Accumulative Constriction by Noradrenaline achieved, and then waveform was recorded. The maximal 30 After sample tension became stable, a piece of waveform response was considered 100%, CaCl constriction percent was recorded, noradrenaline (NA) (3x107-6x10 mol/L) age was used as Y axis, negative logarithms of different con was added into the bath tube until maximal response, then centrations were used as X axis to draw dose effective curve waveform was recorded. Then K-H solution was used to flush (data were expressed in meant-SEM (--), P-0.05, the sample repeatedly, after balanced for 1 hour, compound **P<0.01, n=5), as shown in FIG. 23. 35 II-31 (3x10 mol/L) was added. NA (3x107-3x10" mol/L) From FIG. 23, dose effective curve of CaCl was obviously was also added by using the same method 20 min later. The moved to right in parallel, with maximal response nearly maximal response was considered 100%, NA constriction unchanged. Percentages at different concentrations were sta tistically analyzed, most P values <0.01, Suggesting signifi percentage was used as Y axis, negative logarithms of differ cant difference. The PA value was 6.99+0.05 for amlodipine 40 ent concentrations were used as X axis to draw dose effective to resist constriction of rabbit aorta by CaCl. curve. The curve was shown in FIG. 10 after compound II-31 6.3 Antagonism of Compound II-2 on Hydroxytryptamine (3x10 mol/L) was added. Dose effective curve of NA was (5-HT) Receptor Agonist of Vascular Smooth Muscle of Rab obviously moved to right in parallel, with maximal response bits nearly unchanged. Percentages at different concentrations After sample tension became stable, a piece of waveform 45 were statistically analyzed by using t test, most P values was recorded, 5-HT (107-3x10" mol/L) was added into the <0.01, suggesting significant difference. The PA value was bath tube until maximal response, then waveform was 6.02+0.13 for compound II-31 to resist constriction of rabbit recorded. Then K-H solution was used to flush the sample aorta by NA. repeatedly, after balanced for 1.5 hour, compound II-2 50 1.1.2 Effects of Positive Reference Drug Doxazosin on (3x10 mol/L) was added. 5-HT was also added by using the Dose Effective Curve of Accumulative Constriction by Nora same method 20 min later. The maximal response was con drenaline sidered 100%, 5-HT constriction percentage was used as Y Based on the last step, K-H solution was used to flush the axis, negative logarithms of different concentrations of 5-HT sample repeatedly, after balanced for 1 hour, doxazosin (107 were used as X axis to draw dose effective curve. The curve 55 mol/L) was added. NA was also added by using the same was shown in FIG.7 after compound II-2(3x10" mol/L) was method 15 min later. The maximal response was considered added. Dose effective curve of 5-HT was obviously moved to 100%. NA constriction percentage was used as Y axis, nega right in parallel, with maximal response nearly unchanged. tive logarithms of different concentrations of NA (3x10'-3x Percentages at different concentrations were statistically ana 10 mol/L) were used as X axis to draw dose effective curve. lyzed, P value <0.01, suggesting significant difference. The 60 The curve was shown in FIG. 12 after doxazosin (107 mol/L) PA value was 5.71+0.08 for compound II-2 to resist constric was added. Dose effective curve of NA was obviously moved tion of rabbit aorta by 5-HT. to right in parallel, with maximal response nearly unchanged. 6.4 Antagonism of Compound II-85 on 5-HT Receptor Percentages at different concentrations were statistically ana Agonist of Vascular Smooth Muscle of Rabbits 65 lyzed, most P values <0.01, Suggesting significant difference. After sample tension became stable, a piece of waveform The PA value was 7.76+0.24 for positive drug doxazosin to was recorded, 5-HT (10-10 mol/L) was added into the resist constriction of rabbit aorta by NA. US 9,415,047 B2 117 118 Statistical t test showed that, for PA values of compound 1.3 Antagonism of Compound II-31 on Hydrox II-31 and positive drug doxazosin against NA, P-0.01, Sug ytryptamine (5-HT) Receptor Agonist of Vascular Smooth gesting very significant difference between them, which Muscle of Rabbits meant that compound II-31 had weaker resisting effects After sample tension became stable, a piece of waveform against a receptor agonist than doxazosin. was recorded, 5-HT (10-3x10" mol/L) was added into the 1.2 Antagonism of Compound II-31 on Calcium Channel bath tube until maximal response, then waveform was (Ca") of Vascular Smooth Muscle of Rabbits recorded. Then K-H solution was used to flush the sample 1.2.1 Effects of Compound II-31 on Dose Effect Curve of repeatedly, after balanced for 1.5 hour, compound II-31 CaCl on Accumulative Constriction of Rabbit Blood Vessels (3x10" mol/L) was added. 5-HT was also added by using the After sample tension became stable, calcium K-H solution 10 same method 20 min later. The maximal response was con was used to flush the sample for 3 times, and incubated with sidered 100%, 5-HT constriction percentage was used as Y calcium free K-H solution for 40 min. Calcium free high axis, negative logarithms of different concentrations of 5-HT kalium solution was added into the sample for depolarization were used as X axis to draw dose effective curve. The curve 15 was shown in FIG. 14 after compound II-31 (3x10 mol/L) for 20 min, then CaCl (10-3x10° mol/L) was added into was added. Dose effective curve of 5-HT was obviously bathtube till maximal response was achieved, and then wave moved to right in parallel, with maximal response nearly form was recorded. Then K-H solution was used to flush the unchanged. Percentages at different concentrations were sta sample repeatedly, the K-H solution was changed for every 20 tistically analyzed, P value <0.01, Suggesting significant dif min, and the sample was balanced for 60 min. After sample ference. The PA value was 6.726+0.089 for compound II-31 tension became stable, calcium K-H Solution was used to to resist constriction of rabbit aorta by 5-HT. flush the sample for 3 times, and incubated with calcium free Example 137 K-H solution for 40 min. Calcium free high kalium solution was added into the sample for depolarization for 20 min, then Experiment on Acute Toxicity of Compound II-2 compound II-31 (10 mol/L) was added into bath tube, 25 which was incubated for 20 min, then CaCl (10-3x10' Kunming mice (Experimental Animal Center, China Medi mol/L) was added by using the same method till maximal cal University) were used, half male and half female, 18-22g, response was achieved, and then waveform was recorded. brief probability unit method was used to test the acute tox icity of the compound. The compound LDso for gastric gav The maximal response was considered 100%, CaCl constric 30 age was 361.88 mg/kg (95% confidential interval 302.96 tion percentage was used as Y axis, negative logarithms of 420.80 mg/kg). different concentrations were used as X axis to draw dose effective curve. The curve was shown in FIG. 13 after com Experiment on Acute Toxicity of Compound II-85 pound II-31 (10 mol/L) was added. Dose effective curve of 35 CaCl was obviously moved to right in parallel, with maximal Ten Kunming mice (Experimental Animal Center, China response nearly unchanged. Percentages at different concen Medical University) were used, half male and half female, 18-22 g, brief probability unit method was used to test the trations were statistically analyzed, most P values <0.01, acute toxicity of the compound. The compound LDso was Suggesting significant difference. The PA value was 221.72 mg/kg (95% confidential interval 204.11-239.33 6.56+0.032 for compound II-31 to resist constriction of rabbit 40 mg/kg) for gastric gavage, and 108.32 mg/kg (95% confiden aorta by CaCl. tial interval 102.41-114.23 mg/kg) for intraabdominal injec 1.2.2 Effects of Positive Reference Drug Amlodipine on tion. Dose Effective Curve of Accumulative Constriction by CaCl Example 138 Based on the last step, K-H solution was used to flush the 45 sample repeatedly, the K-H solution was changed for every 20 Marrow Micronucleus Test of Compound II-2 and min, and the sample was balanced for 60 min. After sample II-85 on Mice tension became stable, calcium K-H Solution was used to flush the sample for 3 times, and incubated with calcium free Ten Kunming mice (Experimental Animal Center, China K-H solution for 40 min. Calcium free high kalium solution 50 Medical University), half male and half female, were used. was added into the sample for depolarization for 20 min, then Compound II-2 was administered via gastric gavage at 120 amlodipine (107 mol/L) was added into bathtube, which was mg/kg/day, and II-85 was administered to 74 mg/kg/day. The drug was administered for 4 days continuously. Marrow incubated for 15 min, then CaCl (10-3x10° mol/L) was micronucleus test was performed on day 5. added by using the same method till maximal response was 55 Mice in positive control group were administered cyclo achieved, and then waveform was recorded. The maximal phosphane at 60 mg/kg/day. Mice in negative control group response was considered 100%, CaCl constriction percent were administered normal saline at 0.1 ml/10 g/day. The drug age was used as Y axis, negative logarithms of different con was administered for 4 days continuously. Marrow micro centrations were used as X axis to draw dose effective curve. nucleus test was performed on day 5. The curve was shown in FIG. 13 after amlodipine (10 60 The mice were killed by Snapping neck, and then femur and mol/L) was added. Dose effective curve of CaCl was obvi Sternum were excised, with blood and muscles removed, epi ously moved to right in parallel, with maximal response physis cut off. Then marrow from Sternum was Squeezed to nearly unchanged. Percentages at different concentrations clean glass.slide with bovine serum by using a pair of hemo were statistically analyzed, most P values <0.01, Suggesting 65 static forceps, or marrow on femur was flushed directly with significant difference. The PA value was 7.51+0.288 for bovine serum onto clean glassslide. Then the Substance on amlodipine to resist constriction of rabbit aorta by CaCl. glass slide were mixed well and Smeared. Then the prepared US 9,415,047 B2 119 120 and dried marrow Smears were put into staining tank with polychromatic erythrocytes in blank group, micronuclei cells methanol, fixed for 15 min, picked out to dry in the open air. accounted for 1%; in 1000 polychromatic erythrocytes in After marrow smears were dried, it would be stained in cyclophosphane group, micronuclei cells accounted for 12%. freshly prepared Giemsa solution (1 volume of Giemsa stock The results suggested that, compound II-85 had negative mar solution and 9 volumes of pH 6.8 phosphate buffer) for 10 row micronucleus test. min, staining solution was flushed away with little stream. Then the smear was dried in the open air and observed under Example 139 a microscope. Experimental Results Suggested that: Effects of Compound II-2 on Blood Pressure of SD In 1000 polychromatic erythrocytes in compound II-2 10 Rats group, micronuclei cells accounted for 2.0+0.333%; in 1000 polychromatic erythrocytes in blank group, micronuclei cells Four SD rats were anesthetized with urethane (1.25 accounted for 1%; in 1000 polychromatic erythrocytes in mg/kg), after vital signs became stable, blood pressure was cyclophosphane group, micronuclei cells accounted for 12%. measured by common carotid intubation. After blood pres The results suggested that, compound II-2 had negative mar 15 Sure became stable, compound II-2 was administered at 4.0 row micronucleus test. mg/kg via gastric gavage, blood pressure changes with time In 1000 polychromatic erythrocytes in compound II-85 were observed and recorded. The experimental results were group, micronuclei cells accounted for 2.5+0.373%; in 1000 shown in table 4, 5 and 6. TABLE 4

effects of compound II-2 on diastolic blood pressure (DBP, mmHg) of rats anesthetized with urethane (n = 4)

Drug dose Post dosing (min)

Groups (mg/kg) O 15 30 60 90 120 150 18O 210

Compound 4.0 75.92 62.21 61.84 58.04 53.86 69.10 - 70.79 71.08 - 75.26 II-2 26.19 19.99** 24.65** 18.49** 20.22** 27.71** 27.81* 29.22* 33.42

Note: *P<0.05, **P.<0.01

TABLE 5 effects of compound II-2 on systolic blood pressure (SBP, mmHg) of rats anesthetized with urethane

Drug dose Post dosing (min Groups (mg/kg) O 15 30 60 90 120 150 18O 210 Compound 4.0 122.66 95.77 99.88 - 102.22 98.71 - 111.16 11182 112.34 - 115.12 II-2 20.73 16.29** 22.77:8: 16.29** 13.68** 20.37** 15.75** 15.26** 18.81:

Note: *P<0.05, **P<0.01

TABLE 6 effects of compound II-2 on mean artery pressure (MAP, mmHg) of rats anesthetized with urethane

Drug dose Post dosing (min Groups (mg/kg) O 15 30 60 90 120 150 18O 210 Compound 4.0 91.50 7340 74.52 72.77 68.81 - 83.12 - 84.47 - 84.83 88.54 II-2 24.15 17.80** 23.67** 17.52** 17.92** 25.17** 23.17** 24.50** 28.238

Note: *P<0.05, US 9,415,047 B2 121 122 Experimental results indicated that, compound II-2 had Example 140 obvious hypotensive effects in SD rats anesthetized with ure thane (1.25 mg/kg), and blood pressures recovered to those Effects of Compound II-85 on Blood Pressure of SD before drug dosing 3.5 hours later. Rats In Summary, the above results indicated that, in animal experiments in vitro, compound II-2 had obvious relaxing effects of vascular Smooth muscle. Compound II-2 had equal Five SD rats (provided by Experimental Animal Center, resisting effects to doxazosin on adrenalinea receptor, and the PA value was 7.37+0.08 for compound II to resist noradrena China Medical University) were anesthetized with urethane line (NA), 7.52+0.04 for doxazosin to resist NA, 5.61+0.04 10 (1.25 mg/kg), after vital signs became stable, blood pressure for compound II-2 to resist CaCl2, and 5.71+0.08 to resist was measured by common carotid intubation. After blood 5-HT. In in vivo bulk testing on rats, compound II-2 showed pressure became stable, compound II-85 was administered at good hypotensive effects, good oral absorption, mild toxicity, great therapeutic index, negative marrow micronucleus test, 1.5 mg/kg via gastric gavage, blood pressure changes with with protential value in development of multiple target is time were observed and recorded. The experimental results vasodilative drugs, especially as new hypotensive drugs. were shown in table 7, 8 and 9. TABLE 7

effects of compound II-85 on diastolic blood pressure (DBP, mmHg) of rats anesthetized with urethane (n = 5)

Drug dose Post dosing (min)

Groups (mg/kg) O 15 30 60 90 120 150 18O 210 240

Compound 1.5 103.06 75.36 68.68 - 75.49. 83.32 83.71 - 95.06 - 86.22 97.96 - 102.84 II-85 12.20 23.59* 21.71** 15.19** 23.28* 23.71: 2003 23.24. 21.62. 20.23

Note: *P < 0.05, **P<0.01

TABLE 8 effects of compound II-85 on systolic blood pressure (SBP, mmHg) of rats anesthetized with urethane (n = 5)

Dose Post dosing (min)

Groups (mg/kg) O 15 30 60 90 120 150 18O 210 240

Compound 1.5 143.77 - 125.22 115.90 - 122.75 12548 h 122.89 136.08 - 127.90 - 137.13 - 142.19 II-85 12.69 16.24** 18.14** 1429* 22.07: 21.94 12.15 1981 1948 17.04

Note:

TABLE 9 effects of compound II-85 on mean artery pressure (MAP, mmHg) of rats anesthetized with urethane

Drug dose Post dosing (min

Groups (mg/kg) O 15 30 60 90 120 150 18O 210 240 Compound 1.5 116.63 91.98 84.42 91.24 97.37 96.77 108.73 100.11 111.02 115.95 II-85 12.09 20.84* 20.26** 1481** 21.70% 22.13% 16.96 21.82 20.71 18.84

Note: *P < 0.05, US 9,415,047 B2 123 124 Experimental results indicated that, compound II-85 had when R are groups for poly Substitution, each R is inde obvious hypotensive effects in SD rats anesthetized with ure pendently selected from the group consisting of H, halo thane (1.25 mg/kg), and blood pressures recovered to those gen, CN, C-C alkyl, C-C alkoxyl, CHO, CO(C-C, before drug dosing 6 hours later. alkyl), COO(C-C alkyl), COOH, NO, NH, NH(C- In Summary, the above results indicated that, in animal C alkyl), N(C-C alkyl), SH, S(C-C alkyl). —S(O) experiments in vitro, compound I-85 had obvious relaxing (C-C alkyl). —S(O)H or —S(O) (C-C alkyl), each effects of vascular smooth muscle. Compound II-85 had of which alkyl moiety is optionally substituted with one stronger antagonism against a receptor of adrenaline com or more halogen atoms; pared to doxazosin, and its PA value against PE was A, B and X each independently represents CH or N: 8.62+0.11. The PA value was 6.10+0.13 for compound II-85 10 to resist CaCl. Compound II-85 had stronger antagonism R represents H, halogen, CN, C-C alkyl, C-C alkoxyl, against 5-HT2 receptor, and its PA value against 5-HT was CHO, CO(C-C alkyl), COO(C-C alkyl), COOH, 9.06+0.07. Therefore, compound II-85 can achieve triple NO, NH, NH(C-C alkyl), N(C-C alkyl), SH, effects including obvious C. receptor resistance, Ca" chan S(C-C alkyl). —S(O)(C-C alkyl). —S(O)H or nel blockade, and 5-HT2 receptor resistance, thus achieve 15 —S(O)(C-C alkyl), each of which alkyl moiety is higher efficacy or less side effects compared to single and optionally Substituted with one or more halogenatoms; combined medication to single target drug. In in vivo bulk when R are groups for poly Substitution group, each R testing on rats, compound II-85 showed good hypotensive is independently selected from the group consisting of effects, good oral absorption, mild toxicity, great therapeutic H, halogen, CN, C-C alkyl, C-C alkoxyl, CHO, index, and negative marrow micronucleus test. CO(C-C alkyl), COO(C-C alkyl), COOH, NO, NH, NH(C-C alkyl), N(C-C alkyl), SH, S(C-C, REFERENCES alkyl). —S(O)(C-C alkyl). —S(O)H or —S(O)(C- Calkyl), each of which alkyl moiety is optionally Sub 1. Internal Medicine, Ye Rengao, Lu Zaiying, ed. People's stituted with one or more halogen atoms; and Medical Publishing House, version 6, 2007 April. 25 Y represents a saturated or unsaturated, straight or 2. Pharmacology, Li Rui, ed. People's Medical Publishing branched, Cs hydrocarbon chain optionally substituted House, version 6, 2007 August. with one or more halogen atoms, in which one or more 3. Sanders-Bush E. Mayer S. E. 5-Hydroxytryptamine (sero carbonatoms are optionally replaced with heteroatom(s) tonin): Receptor agonists and antagonists. Selected from oxygen, Sulfur, and nitrogen. In: Brunton L. L. Lazo J. S. Parker K L (eds). Goodman & 30 Gilman's The Pharmacological Basis of Therapeutics 2. The method of claim 1, wherein the pharmaceutically (11th ed), Philadelphia: The McGraw-Hill Companies, acceptable salt is hydrochloride salt, hydrobromide salt, sul 2006:158 phate salt, trifluoroacetate salt, mesylate salt, tartrate salt, 4. Hoyer D. Clarke DE, Fozard J. R. et al. Pharmacol Rev. malate salt, Succinate salt, maleate, citrate salt, phosphate 1994, 46(2): 158 35 salt, lactate salt, pyruvate salt, acetate salt, fumarate salt, 5. Martin G. R. 5-Hydroxytryptamine receptors. In: The oxaloacetate salt, esylate salt, oxalate salt, besylate salt or IUPHAR Committee on Receptor Nomenclature and Drug isethionate salt. Classification (ed), The IUPHAR Compendium of Recep 3. The method of claim 1, wherein the aromatic group is tor Characterization and Classification-, London: phenyl, naphthyl, a benzo-fused five-membered or six-mem IUPHAR Media, 1998: p. 167. 40 bered heterocyclic ring containing heteroatom(s) selected from N, S and O, or a five-membered or six-membered unsat The invention claimed is: urated heterocyclic ring. 1. A method of mediating vasodilation in a Subject in need 4. The method of claim 1, wherein R is H. halogen, CN, thereof, said method comprising administering to said subject C-C alkyl, C-C alkoxyl, CHO, CO(C-C alkyl), COO a pharmaceutical composition effective for mediating said 45 (C-C alkyl) or COOH, each of whichalkyl moiety is option vasodilation, said pharmaceutical composition comprising a ally Substituted with one or more halogen atoms. compound of formula (I) or a pharmaceutically acceptable 5. The method of claim 1, wherein R is H. halogen, CN, salt thereof: C-C alkyl, C-C alkoxyl, CHO, CO(C-C alkyl), COO (C-C alkyl), COOH or NO, each of which alkyl moiety is 50 (I) optionally substituted with one or more halogen atoms. 6. The method of claim 1, wherein Y is an unsubstituted saturated Cls hydrocarbon group, or an unsubstituted Satu rated Cls hydrocarbon group in which one carbon atom is replaced with oxygen or Sulfur. 55 7. The method of claim 1, wherein both A and B represent N. 8. The method of claim 1, wherein wherein: R is an aromatic group mono- or disubstituted with R. R represents an aromatic group or analicyclic group, each 60 wherein of which is mono- or polysubstituted with R, wherein, the aromatic group is phenyl, benzisoxazolyl or ben R represents H. halogen, CN, C-C alkyl, C-C alkoxyl, Zisothiazolyl group; CHO, CO(C-C alkyl), COO(C-C alkyl), COOH, R is H, F, Cl, Br, OCH or CF; NO, NH, NH(C-C alkyl), N(C-C alkyl), SH, A, B and X each independently represents CH or N: S(C-C alkyl). —S(O)(C-C alkyl). —S(O)H or 65 R represents H. F. Cl, Br, CN, CH or OCH; and —S(O)(C-C alkyl), each of which alkyl moiety is Y represents ethylidene, propylidene, butylidene, or eth optionally Substituted with one or more halogen atoms; ylideneoxy.

US 9,415,047 B2 129 130 I-133 N-(4-(1H-benzotriazol-1-yl)propoxyl)-4-(3-trifluo 14. The method of claim 3, wherein the aromatic group is romethylphenyl)piperidine, phenyl or benzisoxazolyl group, wherein (i) the aromatic I-134 N-(4-(1H-benzimidazol-1-yl)propoxyl)-4-(3-trif group is phenyl, and X is N, or (ii) the aromatic group is luoromethylphenyl)piperidine, benzisoxazolyl. X is CH, and A is N. I-135 N-(3-(1H-benzotriazol-1-yl)propyl)-4-(3-(6-meth 15. The method of claim 1, wherein the alicyclic group is ylbenzisoxazolyl)piperidine, cyclopentyl, cyclohexyl, tetrahydrofuryl, piperidyl or piper I-136 N-(3-(1H-benzotriazol-1-yl)propyl)-4-(3-(6-meth aZinyl group. oxylbenzisoxazolyl))piperidine, 16. The method of claim 4, wherein (i) X is N, and R is H. I-137 N-(3-(6-fluoro-1H-benzotriazol-1-yl)propyl)-4-(3- F, Cl, or OCH or (ii) X is CH, and R is H, F, or CF. (6-fluorobenzisoxazolyl))piperidine, 10 I-138 N-(3-(6-chloro-1H-benzotriazol-1-yl)propyl)-4-(3- 17. The method of claim 5, wherein (i) X is N, and R is H. (6-fluorobenzisoxazolyl))piperidine, F C1, CN, CH, or COOCH, or (ii) X is CH, and R is Hor I-139 N-(3-(6-methyl-1H-benzotriazol-1-yl)propyl)-4-(3- OCH. (6-fluorobenzisoxazolyl))piperidine, 18. The method of claim 6, wherein (i)Y is butylidene, and I-140 N-(3-(6-methoxyl-1H-benzotriazol-1-yl)propyl)-4- 15 X is N, or (ii)Y is propylidene, and X is CH. (3-(6-fluorobenzisoxazolyl)piperidine, 19. A method of inducing in a subject in need thereof, I-141 N-(3-(6-formyl-1H-benzotriazol-1-yl)propyl)-4-(3- relaxation of vascular Smooth muscle, said method compris (6-fluorobenzisoxazolyl))piperidine, ing administering to the Subject a pharmaceutical composi I-142 N-(3-(6-methoxylbenzotriazolyl)propyl)-4-(3-ben tion effective for Such relaxation, said pharmaceutical com Zisoxazolyl)piperidine, position comprising a compound of formula (I) or a I-143 N-(2-(1-benzotriazolypethyl)-4-(3-(6-fluoroben pharmaceutically acceptable salt thereof: Zisoxazolyl))piperidine, I-144 N-(4-(1-benzotriazolyl)butyl)-4-(3-(6-fluoroben Zisoxazolyl))piperidine, (I) I-145 N-(4-(6-cyanobenzotriazolyl)butyl)-4-(3-(6-fluo 25 robenzisoxazolyl))piperidine, I-146 N-(4-(6-cyanobenzotriazolyl)butyl)-4-(3-(6-meth oxylbenzisoxazolyl))piperidine, I-147 N-(2-(6-methoxylbenzotriazolyl)ethoxyl)-4-(3-ben Zisoxazolyl)piperidine, 30 I-148 N-(2-(1-benzotriazolyl)ethoxyl)-4-(3-fluoroben Zisoxazolyl)piperidine, wherein: I-149 N-(3-(6-methoxylbenzotriazolyl)propyl)-4-(3-(6- R represents an aromatic group or analicyclic group, each fluorobenzisothiazolyl)piperidine, of which is mono- or polysubstituted with R, wherein, I-150 N-(3-(6-methoxylbenzotriazolyl)propyl)-4-(3-(6- 35 fluorobenzopyrazol))piperidine, R represents H, halogen, CN, C-C alkyl, C-C alkoxyl, I-151 N-(3-(6-methoxylbenzotriazolyl)propyl)-4-(3-(6- CHO, CO(C-C alkyl), COO(C-C alkyl), COOH, fluorobenzofuranyl)piperidine, NO, NH, NH(C-C alkyl), N(C-C alkyl), SH, I-152 N-(4-(1H-benzimidazol-1-yl)butyl)-4-(2-furyl)pip S(C-C alkyl). —S(O)(C-C alkyl). —S(O)H or eridine, 40 —S(O)2(C-C alkyl), each of which alkyl moiety is I-153 N-(4-(1H-benzimidazol-1-yl)butyl)-4-(4-pyridyl) optionally Substituted with one or more halogenatoms; piperidine, when R are groups for poly Substitution, each R is inde I-154 N-(4-(1H-benzimidazol-1-yl)butyl)-4-(2-pyrimidi pendently selected from the group consisting of H, halo nyl)piperidine, gen, CN, C-C alkyl, C-C alkoxyl, CHO, CO(C-C, I-155 N-(4-(1H-benzotriazol-1-yl)butyl)-4-cyclohexylpi 45 alkyl), COO(C-C alkyl), COOH, NO, NH, NH(C- peridine, C alkyl), N(C-C alkyl), SH, S(C-C alkyl). —S(O) I-156 N-(4-(1H-benzotriazol-1-yl)butyl)-4-(1-naphthyl) (C-C alkyl). —S(O)H or —S(O) (C-C alkyl), each piperidine, or of which alkyl moiety is optionally substituted with one I-157 N-(4-(1H-benzotriazol-1-yl)butyl)-4-(2-quinoxali or more halogen atoms; nyl)piperidine. 50 A, B and X each independently represents CH or N: 10. The method of claim 1, wherein the pharmaceutical R represents H, halogen, CN, C-C alkyl, C-C alkoxyl, composition is administered for treatment of hypertension, CHO, CO(C-C alkyl), COO(C-C alkyl), COOH, heart failure, angina pectoris, coronary heart disease, cerebral NO, NH, NH(C-C alkyl), N(C-C alkyl), SH, ischemic disease induced by vascular spasm, myocardial S(C-C alkyl). —S(O)(C-C alkyl). —S(O)H or ischemic disease, shock, renal ischemia, renal dysfunction 55 —S(O)2(C-C alkyl), each of which alkyl moiety is due to renal vascular spasm, or peripheral vascular spasmodic optionally Substituted with one or more halogenatoms; disease. when R are groups for poly Substitution group, each R 11. The method of claim 2, wherein the pharmaceutically is independently selected from the group consisting of acceptable salt contains crystal water. H, halogen, CN, C-C alkyl, C-C alkoxyl, CHO, 12. The method of claim 2, wherein the pharmaceutically 60 CO(C-C alkyl), COO(C-C alkyl), COOH, NO, acceptable salt is hydrochloride salt, hydrobromide salt, sul NH, NH(C-C alkyl), N(C-C alkyl), SH, S(C-C, fate salt or mesylate salt. alkyl). —S(O)(C-C alkyl). —S(O)H or —S(O)(C- 13. The method of claim 3, wherein the aromatic group is Calkyl), each of which alkyl moiety is optionally sub phenyl, naphthyl, benzothiazolyl, benzisothiazolyl, benzox stituted with one or more halogen atoms; and azolyl, benzisoxazolyl, benzimidazoyl, benzopyrazol, ben 65 Y represents a saturated or unsaturated, straight or Zofuryl, benzopyrimidinyl, benzopyridyl, quinoxalinyl, branched, Cs hydrocarbon chain optionally Substituted furyl, pyridyl or pyrimidinyl group. with one or more halogen atoms, in which one or more US 9,415,047 B2 131 132 carbonatoms are optionally replaced with heteroatom(s) Selected from oxygen, Sulfur, and nitrogen, wherein the compound of formula (I) or a pharmaceuti cally acceptable salt thereof is at least one of an O. receptor antagonist, a Ca" channel blocker, and a 5 5-HT2 receptor antagonist. k k k k k UNITED STATES PATENT AND TRADEMARK OFFICE CERTIFICATE OF CORRECTION

PATENT NO. : 9,415,047 B2 Page 1 of 1 APPLICATION NO. : 14/647408 DATED : August 16, 2016 INVENTOR(S) : Yan Zhou et al. It is certified that error appears in the above-identified patent and that said Letters Patent is hereby corrected as shown below:

In The Specification Column 47, line 24: "against a receptors' should be --against O. receptors--.

Column 104, lines 10-11: “of I-2 bubbles/second. Should be --of 1-2 bubbles/second.--.

Column 111, line 21 : “Suizhou hake should be --Suizhou Jiake--.

Column 112, lines 16-17: “of I-2 bubbles/second. Should be --of 1-2 bubbles/second.--. Column 113, line 2: "against a receptor should be --against O. receptor--. Column 113, line 52: "against a receptor should be --against O. receptor--. Column 117, line 5: "against a receptor should be --against O. receptor--. Column 121, line 8: “adrenaline a receptor, should be -adrenaline O. receptor.--.

Signed and Sealed this Eighth Day of November, 2016 74-4-04- 2% 4 Michelle K. Lee Director of the United States Patent and Trademark Office