USOO7718809B2

(12) United States Patent (10) Patent No.: US 7,718,809 B2 Koike et al. (45) Date of Patent: *May 18, 2010

(54) CHROMANE SUBSTITUTED WO WO 2004.054984 T 2004 BENNY PRIVATIVES ASACD OTHER PUBLICATIONS Thomas A. Godwin, Gastrointestinal Diseases, http://edcenter.med. (75) Inventors: Hiroki Koike, Chita-gun (JP); Sachiko cornell.edu/CUMC PathNotes/Gastrointestinal/Gastrointestinal. Sakakibara, Chita-gun (JP) html, Aug. 2004, 51 pages.” International Search Report for PCT/IB2006/001567, 6 pages, Aug. (73) Assignee:rsr rr RaGualia Pharma Inc., Aichi-kenrhi- (JP) Written2006. Opinion by International Searching Authority for PCT/ (*) Notice: Subject to any disclaimer, the term of this IB2006/001567, 6 pages, Dec. 2007. patent is extended or adjusted under 35 * cited by examiner U.S.C. 154(b)(b) by 185 days.ayS Primary Examiner Laura L. Stockton This patent is Subject to a terminal dis- (74) Attorney, Agent, or Firm Scully, Scott, Murphy & claimer. Presser, P.C. (21) Appl. No.: 11/916,671 (57) ABSTRACT 1-1. This invention relates to compounds of the formula (I): or a (22) PCT Filed: Jun. 2, 2006 pharmaceutically acceptable salt thereof, wherein: A, B, X, (86). PCT No.: PCT/B2O06/OO1567 R", R. R. R. Rand R. R. RandR are each as described herein or a pharmaceutically acceptable salt, and composi S371 (c)(1) tions containing Such compounds and the use of Such com (2), (4) Date: Dec. 6, 2007 pounds in the treatment of a condition mediated by acid pump s 9 antagonistic activity Such as, but not limited to, as gastrointes (87) PCT Pub. No.: WO2006/134460 tinal disease, gastroesophageal disease, gastroesophageal reflux disease (GERD), peptic ulcer, gastric ulcer, duodenal PCT Pub. Date: Dec. 21, 2006 ulcer, NSAID-induced ulcers, gastritis, infection of Helico bacter pylori, dyspepsia, functional dyspepsia, Zollinger-El (65) Prior Publication Data lison syndrome, non-erosive reflux disease (NERD), visceral pain, heartburn, nausea, esophagitis, dysphagia, hypersaliva US 2008/0214519 A1 Sep. 4, 2008 tion, airway disorders or asthma. Related U.S. Application Data (60) Provisional application No. 60/690,626, filed on Jun. O (I) 14, 2005. RI R3 \ (51) Int. Cl. NN A6 IK 3/484 (2006.01) l, X-R CO7D 405/2 (2006.01) R R9 N (52) U.S. Cl...... 548/304.4: 514/394 (58) Field of Classification Search ...... 548/304.4 X See application file for complete search history. A. (56) References Cited B R5 U.S. PATENT DOCUMENTS 5,106,862 A 4, 1992 Briving et al...... 514,394 R8 R6 6,465,505 B1 10/2002 Amin et al...... 514,394 R7 FOREIGN PATENT DOCUMENTS EP O266326 3, 1993 WO WO9747603 12/1997 5 Claims, No Drawings US 7,718,809 B2 1. 2 CHROMANE SUBSTITUTED visceral pain, heartburn, nausea, esophagitis, dysphagia, BENZMIDAZOLE DERVATIVES ASACD hypersalivation, airway disorders or asthma (hereinafter, PUMP ANTAGONSTS referred as “APA Diseases”). The present invention provides a compound of the follow BACKGROUND OF THE INVENTION 5 ing formula (I):

This invention relates to chromane substituted benzimida (I) Zole derivatives. These compounds have selective acid pump inhibitory activity. The present invention also relates to a 10 3 ,' pharmaceutical composition, method of treatment and use, R N N comprising the above derivatives for the treatment of disease R2 conditions mediated by acid pump modulating activity; in X particular acid pump inhibitory activity. R9 N It has been well established that proton pump inhibitors 15 X (PPIs) are prodrugs that undergo an acid-catalyzed chemical A. rearrangement that permits them to inhibit H/K"-ATPase by B R5 covalently biding to its Cystein residues (Sachs, G. et. al., Digestive Diseases and Sciences, 1995, 40, 3S-23S: Sachset. al., Annu Rev Pharmacol Toxicol, 1995, 35,277-305.). How R8 R6 ever, unlike PPIs, acid pump antagonists inhibit acid secretion via reversible potassium-competitive inhibition of H/K"- ATPase. SCH28080 is one of Such reversible inhibitors and has been studied extensively. Other newer agents (revapra Zan, Soraprazan, AZD-0865 and CS-526) have entered in 25 or a pharmaceutically acceptable salt thereof, wherein: clinical trials confirming their efficacy in human (Pope, A.; -A-B-represents —O—CH2—, —S—CH2—, —CH2 Parsons, M., Trends in Pharmacological Sciences, 1993, 14, O— or —CH2—S-; 323-5; Vakil, N., Alimentary Pharmacology and Therapeu X represents an oxygen atom or NH; tics, 2004, 19, 1041-1049). In general, acid pump antagonists R" and R independently represent a C-C alkyl group are found to be useful for the treatment of a variety of dis 30 being unsubstituted or substituted with 1 to 2 substitu eases, including gastrointestinal disease, gastroesophageal ents independently selected from the group consisting of disease, gastroesophageal reflux disease (GERD), peptic a hydroxy group and a C-C alkoxy group: ulcer, gastric ulcer, duodenal ulcer, non-steroidal anti-inflam Rand R' independently represent a hydrogen atom, a matory drug(NSAID)-induced ulcers, gastritis, infection of C-C alkyl group or a C-C-7 cycloalkyl group, said Helicobacter pylori, dyspepsia, functional dyspepsia, 35 C-C alkyl group and said C-C, cycloalkyl group Zollinger-Ellison syndrome, non-erosive reflux disease being unsubstituted or substituted with 1 to 3 substitu (NERD), visceral pain, heartburn, nausea, esophagitis, dys ents independently selected from the group consisting of phagia, hypersalivation, airway disorders or asthma(Kiljan a halogenatom, a hydroxy group, a C-C alkoxy group der, Toni O, American Journal of Medicine, 2003, 115(Suppl. and a C-C, cycloalkyl group; or R and R' taken 3A), 65S-71S.). 40 together with the nitrogen atom to which they are WOO4/054984 discloses compounds reported to be acid attached form a 4 to 6 membered heterocyclic group pump antagonists. They refer to certain compounds having a being unsubstituted or substituted with 1 to 3 substitu benzimidazole structure. ents selected from the group consisting of a hydroxy There is a need to provide new acid pump antagonists that 45 group, a C-C alkyl group, a C-C acyl group and a are good drug candidates and address unmet needs by PPIs for hydroxy-C-C alkyl group; treating diseases. In particular, preferred compounds should R. R. R. and R independently represent a hydrogen bind potently to the acid pump whilst showing little affinity atom, a halogen atom, a hydroxy group, a C-C alkyl for other receptors and show functional activity as inhibitors group or a C-C alkoxy group; and of acid-secretion in stomach. They should be well absorbed R represents a hydrogenatom, a hydroxy group or a C-C, from the gastrointestinal tract, be metabolically stable and 50 alkoxy group. possess favorable pharmacokinetic properties. They should Also, the present invention provides the use of a compound be non-toxic. Furthermore, the ideal drug candidate will exist of formula (I) or a pharmaceutically acceptable salt thereof, in a physical form that is stable, non-hygroscopic and easily each as described herein, for the manufacture of a medica formulated. 55 ment for the treatment of a condition mediated by acid pump modulating activity; in particular, acid pump inhibitory activ SUMMARY OF THE INVENTION ity. Preferably, the present invention also provides the use of a In this invention, it has now been found out that the new compound of formula (I) or a pharmaceutically acceptable class of compounds having a chromane moiety show acid 60 salt thereof, each as described herein, for the manufacture of pump inhibitory activity and favorable properties as drug a medicament for the treatment of diseases selected from APA candidates, and thus are useful for the treatment of disease Diseases. conditions mediated by acid pump inhibitory activity Such as Also, the present invention provides a pharmaceutical gastrointestinal disease, gastroesophageal disease, GERD. composition comprising a compound of formula (I) or a phar peptic ulcer, gastric ulcer, duodenal ulcer, NSAID-induced 65 maceutically acceptable salt thereof, each as described ulcers, gastritis, infection of Helicobacter pylori, dyspepsia, herein, together with a pharmaceutically acceptable carrier functional dyspepsia, Zollinger-Ellison syndrome, NERD, for said compound. US 7,718,809 B2 3 4 Also, the present invention provides a pharmaceutical propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy and tert composition comprising a compound of formula (I) or a phar butoxy group. Of these, a C-C alkoxy group is preferred; a maceutically acceptable salt thereof, each as described methoxy group is more preferred. herein, together with a pharmaceutically acceptable carrier Where R. R. R. Rand the substituent of R and Rare for said compound and other pharmacologically active the halogenatom, this may be a fluorine, chlorine, bromine or agent(s). iodine atom. Of these, a fluorine atom is preferred. Further, the present invention provides a method of treat Where-A-B is —O CH or —S CH , -A-cor ment of a condition mediated by acid pump inhibitory activ responds —O— or —S—and —B- corresponds —CH2—. ity, in a mammalian Subject including a human, which com Where-A-B is —CH O— or —CH, S -A-cor prises administering to a mammal in need of such treatment a 10 responds —CH2—and —B- corresponds—O— or —S—. therapeutically effective amount of a compound of formula The term “treating and “treatment’, as used herein, refers (I) or a pharmaceutically acceptable salt thereof, each as to curative, palliative and prophylactic treatment, including described herein. reversing, alleviating, inhibiting the progress of, or prevent Examples of conditions mediated by acid pump inhibitory ing the disorder or condition to which Such term applies, or activity include, but are not limited to, APA Diseases. 15 one or more symptoms of Such disorder or condition. The compounds of the present invention may show good Preferred compound of the present invention are those bioavailability, less toxicity, good absorption, distribution, compounds of formula (I) or a pharmaceutically acceptable good solubility, less protein binding affinity other than acid salt thereof, each as described herein, in which: pump, less drug-drug interaction, and good metabolic stabil (A) -A-B - is —O—CH2—, —S-CH2—, —CH2—O— or ity. —CH2 S : X is an oxygen atom; R' and R are inde pendently a C-C alkyl group being unsubstituted or sub DETAILED DESCRIPTION OF THE INVENTION stituted with 1 to 2 substituents independently selected from the group consisting of a hydroxy group and a C-C, In the compounds of the present invention: alkoxy group; R and R are independently a hydrogen Where R',R,R,R,R,R,R7 and Rare the C-C alkyl 25 atom, a C-C alkyl group or a C-C cycloalkyl group, said group, this C-C alkyl group may be a straight or branched C-C alkyl group and said C-C-7 cycloalkyl group being chain group having one to four carbon atoms, and examples unsubstituted or substituted with 1 to 3 substituents inde include, but are not limited to, a methyl, ethyl, propyl, iso pendently selected from the group consisting of a halogen propyl, butyl, isobutyl, sec-butyl and tert-butyl group. Of atom, a hydroxy group, a C-C alkoxy group and a C-C, these, a C-C alkyl group is preferred; a C-C alkyl group is 30 cycloalkyl group; or R and R taken together with the further preferred; a methyl group is particularly preferred. nitrogenatom to which they are attached formanaZetidinyl Where R and R and the subsistent of R and Rare the group, a pyrrolidinyl group or a piperazinyl group, said C-C cycloalkyl group, this represents a cycloalkyl group aZetidinyl group, said pyrrolidinyl group and said piperazi having three to seven carbon atoms, and examples include a nyl group being unsubstituted or substituted with 1 to 3 cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cyclo 35 Substituents selected from the group consisting of a heptyl group. Of these, a C-C cycloalkyl group is preferred; hydroxy group, a C-C alkyl group, a C-C acyl group a cyclopropyl group is more preferred. and a hydroxy-C-C alkyl group; RandR are indepen Where RandR' taken together with the nitrogenatom to dently a hydrogenatom, a halogenatom, a hydroxy group, which they are attached form a 4 to 6 membered heterocyclic a C-C alkyl group or a C-C alkoxy group; and R. R group, this 4 to 6 membered heterocyclic group represents a 40 and Rare each a hydrogen atom. saturated heterocyclic group having three to five ring atoms (B)-A-B - is —O—CH2— or —CH2—O—; X is an oxygen selected from carbon atom, nitrogen atom, Sulfur atom and atom; R' and Rare independently a C-C alkyl group; R oxygenatom other than the said nitrogenatom, and examples and Rare independently a hydrogen atom, a C-C alkyl include, but are not limited to, a pyrrolidinyl, imidazolidinyl, group being unsubstituted or Substituted with a hydroxy pyrazolidinyl, piperidinyl, piperazinyl, morpholino, thiomor 45 group; or R and R taken together with the nitrogenatom pholino, azetidinyl. Of these, a pyrrolidinyl, azetidinyl and to which they are attached form an aZetidinyl group, a piperazinyl group are more preferred. pyrrolidinyl group or a piperazinyl group, said aZetidinyl Where the substituent of the 4 to 6 membered heterocyclic group, said pyrrolidinyl group and said piperazinyl group group is the hydroxy-C-C alkyl group, this represents the being unsubstituted or substituted with 1 to 2 substituents said C-C alkyl group Substituted with a hydroxy group, and 50 selected from the group consisting of a hydroxy group, a examples include, but are not limited to, a hydroxymethyl, C-C alkyl group, a C-C acyl group and a hydroxy-C- 2-hydroxyethyl, 1-hydroxyethyl, 3-hydroxypropyl, 2-hy C. alkyl group; R and Rare independently a hydrogen droxypropyl, 2-hydroxy-1-methylethyl, 4-hydroxybuthyl, atom, a halogen atom or a C-C alkyl group; and R. R. 3-hydroxybuthyl, 2-hydroxybuthyl, 3-hydroxy-2-methylpro and Rare each a hydrogen atom. pyl and 3-hydroxy-1-methylpropyl group. Of these, a 55 (C) -A-B - is —O—CH2—, —S-CH2—, —CH2—O— or hydroxy-C-C alky group is preferred; a hydroxymethyl —CH S : X is an oxygen atom; R' and R are inde group is more preferred. pendently a C-C alkyl group; R and R are indepen Where the substituent of the 4 to 6 membered heterocyclic dently a C-C alkyl group being unsubstituted or Substi group is the C-C acyl group, this represents a carbonyl tuted with 1 to 3 substituents independently selected from group Substituted with the said C-C alkyl group, and 60 the group consisting of a halogenatom, a hydroxy group, a examples include, but are not limited to, a formyl, acetyl, C-C alkoxy group and a C-C, cycloalkyl group; or R propionyl, butyryl group. Of these, an acetyl group is pre and R' taken together with the nitrogenatom to which they ferred. are attached form a pyrrolidinyl group, an azetidinyl group Where R. R. R. R. Rand the substituents of R. R. R. or a piperazinyl group, said pyrrolidinyl group, said aze and Rare the C1-C4 alkoxy group, this represents an oxygen 65 tidinyl group and said piperazinyl group being unsubsti atom Substituted with the said C-C alkyl group, and tuted or substituted with 1 to 3 substituents selected from examples include, but are not limited to, a methoxy, ethoxy, the group consisting of a hydroxy group, a C-C alkyl US 7,718,809 B2 5 6 group, a C-C acyl group and a hydroxy-C-C alkyl stituted or substituted with 1 to 3 substituents indepen group; R. and Rare independently a hydrogen atom, a dently selected from the group consisting of a hydroxy halogen atom, a hydroxy group, a C-C alkyl group or a group and a C-C alkoxy group; R is a hydrogen atom or C-C alkoxy group; and R. RandR are eachahydrogen a halogenatom; Risahalogenatom; and R. RandR are atOm. each a hydrogen atom. (D)-A-B - is —O—CH2— or —CH2—O : X is an oxygen (J)-A-B is —CH2—O : X is an oxygenatom; R', R. R. atom; R and Rare independently a C-C alkyl group; R and R are each a methyl group; R and R7 are indepen and Rare independently a C-C alkyl group being unsub dently a hydrogenatom, a halogenatom or a methyl group; stituted or substituted with 1 to 3 substituents indepen and R. RandR are each a hydrogen atom. dently selected from the group consisting of a hydroxy 10 (K)-A-B is —CH2—O : X is an oxygen atom; R' and group and a C-C alkoxy group; or R and R' taken Rare each a methyl group; R and R are each a methyl together with the nitrogenatom to which they are attached group; or Rand R taken together with the nitrogenatom to form a pyrrolidinyl group being unsubstituted or Substi which they are attached form an azetidin-1-yl group, a pyr tuted with 1 to 2 substituents selected from the group rolidin-1-yl group, a 3-hydroxy-pyrrolidin-1-yl group or a consisting of a hydroxy group and a hydroxy-C-C alkyl 15 4-acetyl-piperazin-1-yl group; RandR are independently a group; R and R are independently a hydrogen atom, a hydrogenatom, a halogenatom or a methyl group; and R. R halogen atom, a hydroxy group a C-C alkyl group or a and Rare each a hydrogen atom. C-C alkoxy group; and R. RandR are eachahydrogen Preferred classes of compound of the present invention are atOm. those compounds of formula (I) or a pharmaceutically accept (E)-A-B - is —O—CH2— or—CH2—O—; X is an oxygen able salt thereof, each as described herein, in which: atom; R and Rare independently a C-C alkyl group; R (a) -A-B - is —O—CH2—, or —CH2—O—: and Rare independently a C-C alkyl group being unsub (b) R' is a C-C alkyl group; stituted or substituted with 1 to 3 substituents indepen (c) R' is a C-C alkyl group; dently selected from the group consisting of a hydroxy (d) R' is a methyl group; group and a C-C alkoxy group; or R and R' taken 25 (e) R is a C-C alkyl group; together with the nitrogenatom to which they are attached (f) R is a C-C alkyl group; form a pyrrolidinyl group being unsubstituted or Substi (g) R is a methyl group; tuted with 1 to 3 substituents selected from the group (h) R is a C-C alkyl group being unsubstituted or substi consisting of a hydroxy group and a hydroxy-C-C alkyl tuted with 1 to 2 substituents independently selected from group; R and R are independently a hydrogen atom, a 30 the group consisting of a hydroxy group and a C-C, halogen atom, a hydroxy group or C-C alkyl group; and alkoxy group; R. RandR are each a hydrogenatom. (i) R is a C-C alkyl group being unsubstituted or substi (F)-A-B - is —CH2—O— or —CH2—S : X is an oxygen tuted with a substituent independently selected from the atom; R' and R are independently a C-C alkyl group group consisting of a hydroxy group and a C-C alkoxy being unsubstituted or substituted with 1 to 2 substituents 35 group; independently selected from the group consisting of a (j) R is a C-C alkyl group being unsubstituted or substi hydroxy group and a C-C alkoxy group; R and Rare tuted with 1 to 2 hydroxy groups; independently a C-C alkyl group; or R and R' taken (k) R is a C-C alkyl group being unsubstituted or substi together with the nitrogenatom to which they are attached tuted with a hydroxy group; form a pyrrolidinyl group being unsubstituted or Substi 40 (1) R' is a C-C alkyl group being unsubstituted or substi tuted with 1 to 2 substituents selected from the group tuted with 1 to 2 substituents independently selected from consisting of a hydroxy group and a hydroxy-C-C alkyl the group consisting of a hydroxy group and a C-C, group; R and R are independently a hydrogen atom, a alkoxy group; halogen atom, a hydroxy group or C-C alkyl group; and (m) R is a methyl group or 2-hydroxyethyl group; R. R. and Rare each a hydrogenatom. 45 (n) R' is a hydrogen atom or a C-C alkyl group; (G)-A-B - is —O—CH2— or —CH2—O : X is an oxygen (o) R' is a C-C alkyl group; atom; R' and R are independently a C-C alkyl group (p) R' is a C-C alkyl group; being unsubstituted or substituted with 1 to 2 substituents (q) R' is a methyl group; independently selected from the group consisting of a (r) RandR taken together with the nitrogen atom to which hydroxy group and a C-C alkoxy group; R and Rare 50 they are attached form a 5 to 6 membered heterocyclic independently a C-C alkyl group being unsubstituted or group optionally having a further a nitrogen atom, said substituted with 1 to 3 substituents independently selected heterocyclic group being unsubstituted or Substituted with from the group consisting of a hydroxy group and a C-C, 1 to 3 Substituents selected from the group consisting of a alkoxy group; R is a C-C alkyl group; and R. R. Rand hydroxy group, a C-C alkyl group, a C-C acyl group Rare each a hydrogen atom. 55 and a hydroxy-C-C alkyl group; (H)-A-B - is —O—CH2— or —CH2—O—: X is an oxygen (s) RandR taken together with the nitrogen atom to which atom; RandR are independently a C-C alkyl group; R they are attached form a pyrrolidinyl group, an aZetidinyl and Rare independently a C-C alkyl group being unsub group or a piperazinyl group, said pyrrolidinyl group, said stituted or substituted with 1 to 3 substituents indepen aZetidinyl group and said piperazinyl group being unsub dently selected from the group consisting of a halogen 60 stituted or substituted with 1 to 3 substituents selected from atom, a hydroxy group, a C-C alkoxy group and a C-C, the group consisting of a hydroxy group, a C-C alkyl cycloalkyl group; R is a hydrogenatom or a halogenatom; group, a C-C acyl group and a hydroxy-C-C alkyl R’ is a halogenatom; and R. Rand Rare each a hydro group; gen atom. (t) RandR' taken together with the nitrogen atom to which (I)-A-B - is —O—CH2—, or—CH2—O—; X is an oxygen 65 they are attached form a pyrrolidinyl group or a piperazinyl atom; R and Rare independently a C-C alkyl group; R group, said pyrrolidinyl group and said piperazinyl group and Rare independently a C-C alkyl group being unsub being unsubstituted or substituted with 1 to 3 substituents US 7,718,809 B2 7 8 Selected from the group consisting of a hydroxy group, a phosphate/dihydrogen phosphate, pyroglutamate, saccha C-C alkyl group, a C-C acyl group and a hydroxy-C- rate, Stearate. Succinate, tannate, tartrate, tosylate, trifluoro C2 alkyl group; acetate and Xinofoate salts. (u) RandR' taken together with the nitrogenatom to which Suitable base salts are formed from bases which form they are attached form a pyrrolidinyl group being unsub non-toxic salts. Examples include the aluminium, arginine, stituted or substituted with 1 to 3 substituents selected from benZathine, calcium, choline, diethylamine, diolamine, gly the group consisting of a hydroxy group and a hydroxy cine, lysine, magnesium, meglumine, olamine, potassium, C-C alkyl group; Sodium, tromethamine and Zinc salts. (v) RandR taken together with the nitrogenatom to which For a review on suitable salts, see "Handbook of Pharma they are attached form a pyrrolidinyl group being unsub 10 ceutical Salts: Properties, Selection, and Use' by Stahl and stituted or substituted with a substituent selected from the Wermuth (Wiley-VCH. Weinheim, Germany, 2002). A phar group consisting of a hydroxy group and a 2-hydroxyethyl maceutically acceptable salt of a compound of formula (I) group; may be readily prepared by mixing together solutions of the (w) RandR' taken together with the nitrogenatom to which compound of formula (I) and the desired acid or base, as they are attached formanaZetidin-1-yl group, a 3-hydroxy 15 appropriate. The salt may precipitate from solution and be 3-methyl-aZetidin-1-yl group, a pyrrolidin-1-yl group, a collected by filtration or may be recovered by evaporation of 3-hydroxy-pyrrolidin-1-yl group, a 2-hydroxymethyl-pyr the solvent. The degree of ionization in the salt may vary from rolidin-1-yl group, a 4-acetyl-piperazin-1-yl group or a completely ionized to almost non-ionized. 4-methyl-piperazin-1-yl group; Pharmaceutically acceptable salts of the compounds of (x) RandR' taken together with the nitrogenatom to which formula (I) thereof include both unsolvated and solvated they are attached formanaZetidin-1-yl group, a 3-hydroxy forms. The term "solvate” is used hereinto describe a molecu pyrrolidin-1-yl group or a 4-acetyl-piperazin-1-yl group; lar complex comprising a compound of the invention and one (y) R is a hydrogen atom, a halogen atom, a hydroxy group, or more pharmaceutically acceptable solvent molecules, for a C-C alkyl group or a C-C alkoxy group: example, ethanol. The term hydrate is employed when said (Z) R is a hydrogen atom, a halogen atom or a methyl group; 25 solvent is water. (aa) R is a hydroxy group, a C-C alkyl group or a C-C, Pharmaceutically acceptable Solvates in accordance with alkoxy group; the invention include hydrates and solvates wherein the sol (bb) R is a hydrogen atom, a chlorine atom or a fluorine vent of crystallization may be isotopically Substituted, e.g. atom; D.O, de-acetone, d-DMSO. (cc) R is a hydrogen atom or a fluorine atom; 30 The term “compound of the invention” or “compounds of (dd) R is a C-C alkyl group; the invention” refers to, unless indicated otherwise, a com (ee) R is a methyl group; pound of formula (I) as hereinbefore defined, isomers thereof (f) R' is a hydrogen atom or a halogen atom; (including optical, geometric and tautomeric isomers) as (gg) R' is a hydrogen atom; hereinafter defined and isotopically-labeled compounds of (hh) R' is a chlorine atom or a fluorine atom; 35 formula (I). (ii) R' is a fluorine atom; Compounds of formula (I) containing one or more asym (ii) RandR are independently a hydrogen atom or halogen metric carbon atoms can exist as two or more stereoisomers. atom; Where the compound contains a keto moiety, tautomeric (kk) R' and Rare each a hydrogenatom; isomerism (“tautomerism') can occur. It follows that a single (l)R’ is a hydrogenatom, a hydroxy group or a C-C alkoxy 40 compound may exhibit more than one type of isomerism. group; Included within the scope of the present invention are all (mm) R is a hydrogen atom. Stereoisomers, geometric isomers and tautomeric forms of Of these classes of compounds, any combination among the compounds of formula (I), including compounds exhib (a) to (mm) is also preferred. 45 iting more than one type of isomerism, and mixtures of one or One embodiment of the invention provides a compound more thereof. Also included are acid addition or base salts selected from the group consisting of 4-(7-Fluoro-3,4-dihy wherein the counterion is optically active, for example, dro-2H-chromen-4-yl)oxy-N,N,1,2-tetramethyl-1H-benz D-lactate or L-lysine, or racemic, for example, DL-tartrate or imidazole-6-carboxamide: 4-(5,7-Difluoro-3,4-dihydro DL-arginine. 2H-chromen-4-yl)oxy-N,N,1,2-tetramethyl-1H 50 The present invention includes all pharmaceutically benzimidazole-6-carboxamide; N.N.1.2-Tetramethyl-4-(5- acceptable isotopically-labeled compounds of formula (I) methyl-3,4-dihydro-2H-chromen-4-yl)oxy-1H wherein one or more atoms are replaced by atoms having the benzimidazole-6-carboxamide; or a pharmaceutical same atomic number, but an atomic mass or mass number acceptable salt thereof. different from the atomic mass or mass number usually found Pharmaceutically acceptable salts of a compound of for 55 in nature. mula (I) include the acid addition salts and base salts (includ Examples of isotopes suitable for inclusion in the com ing disalts) thereof. pounds of the invention include isotopes of hydrogen, Such as Suitable acid addition salts are formed from acids which *H and H. carbon, such as 'C, 'Cand ''C, chlorine, such as form non-toxic salts. Examples include the acetate, adipate, Cl, fluorine, such as 'F, iodine, such as 'I and 'I, aspartate, benzoate, besylate, bicarbonate/carbonate, bisul 60 nitrogen, such as 'N and 'N, oxygen, such as 'O, ''O and phate? Sulphate, borate, camsylate, citrate, cyclamate, edisy 'O, phosphorus, such as 'P and sulphur, such as S. late, esylate, formate, fumarate, gluceptate, gluconate, glucu Certain isotopically-labeled compounds of formula (I), for ronate, hexafluorophosphate, hibenzate, hydrochloride/ example, those incorporating a radioactive isotope, are useful chloride, hydrobromide/bromide, hydroiodide/iodide, in drug and/or substrate tissue distribution studies. The radio isethionate, lactate, malate, maleate, malonate, mesylate, 65 active isotopes tritium, i.e. H, and carbon-14, i.e. ''C, are methylsulphate, naphthylate, 2-napsylate, nicotinate, nitrate, particularly useful for this purpose in view of their ease of orotate, oxalate, palmitate, pamoate, phosphate/hydrogen incorporation and ready means of detection. US 7,718,809 B2 10 Substitution with heavier isotopes such as deuterium, i.e. fH, may afford certain therapeutic advantages resulting from -continued O greater metabolic stability, for example, increased in vivo R half-life or reduced dosage requirements, and hence may be M preferred in some circumstances. RS N Substitution with positron emitting isotopes, such as 'C, X R2 'F, 'O and 'N, can be useful in Positron Emission Topog R9 N raphy (PET) studies for examining substrate receptor occu X pancy. 10 A Isotopically-labeled compounds of formula (I) can gener B R5 ally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying examples and preparations using an R8 R6 appropriate isotopically-labeled reagents in place of the non 15 labeled reagent previously employed. All of the compounds of the formula (I) can be prepared by (I) the procedures described in the general methods presented below or by the specific methods described in the examples In Reaction Scheme A. R. R. R. R. R. R. R7, R, R, section and the preparations section, or by routine modifica A, B and X are each as defined above; LV is a leaving group; R" is R' as defined above or R' wherein hydroxy group is tions thereof. The present invention also encompasses any protected by a hydroxy-protecting group; R is Ras defined one or more of these processes for preparing the compounds above or R wherein hydroxy group is protected by a of formula (I), in addition to any novel intermediates used 25 hydroxy-protecting group; R is Ras defined above or R therein. wherein hydroxy group is protected by a hydroxy-protecting group; R is R as defined above or R' wherein hydroxy General Synthesis group is protected by a hydroxy-protecting group; R is Ras The compounds of the present invention may be prepared defined above or R wherein hydroxy group is protected by a by a variety of processes well known for the preparation of 30 hydroxy-protecting group; R' is Ras defined above or R compounds of this type, for example as shown in the follow wherein hydroxy group is protected by a hydroxy-protecting ing Method A to D. group; R is R as defined above or R' wherein hydroxy All starting materials in the following general syntheses group is protected by a hydroxy-protecting group; R is Ras may be commercially available or obtained by the following defined above or R wherein hydroxy group is protected by a Method E to G or conventional methods known to those 35 hydroxy-protecting group; and R” is Ras defined above or skilled in the art, such as WO 2000078751 and WO R wherein hydroxy group is protected by a hydroxy-protect 2004.054984 and the disclosures of which are incorporated ing group; and the same shall apply hereinafter. herein by references. The term “leaving group', as used herein, signifies a group capable of being Substituted by nucleophilic groups, such as Method A 40 a hydroxy group or amines and examples of Such leaving This illustrates the preparation of compounds of formula groups include a halogen atom, a alkylsulfonyloxy group, a (I). halogenoalkylsulfonyloxy group and a phenylsulfonyloxy group. Of these, a bromine atom, a chlorine atom, a methyl Sulfonyloxy group, a trifluoromethylsulfonyloxy group and a 45 Reaction Scheme A 4-methylphenylsulfonyloxy group are preferred. O The term "hydroxy-protecting groups', as used herein, R 3. signifies a protecting group capable of being cleaved by vari 3 A ous means to yield a hydroxy group. Such as hydrogenolysis, RS N hydrolysis, electrolysis or photolysis, and Such hydroxy-pro 50 R4a X- R2a + tecting groups are described in Protective Groups in Organic N Synthesis edited by T. W. Greene et al. (John Wiley & Sons, 1999). Such as for example, C-C alkoxycarbonyl, C-C, XH alkylcarbonyl, tri-C-C alkylsilyl or tri-C-C alkylarylsilyl groups, and C-C alkoxy-C-C alkyl groups. Suitable (II) 55 hydroxy-protecting groups include acetyl and tert-butyldim R9a ethylsilyl. Lw A (Step A1) In the Step A1, the compound of formula (I) is prepared by B R5a Step Al 60 nucleophilic substitution with LV of the compound of formula (III), which is commercially available or may be prepared by the method described in WO 2000078751, US 20050038032 R8a R6a or the following Method E to F, with the compound of for R7a mula (II), which is commercially available or may be pre 65 pared by the methods described in WO 2004.054984. The reaction is normally and preferably effected in the presence of solvent. There is no particular restriction on the US 7,718,809 B2 11 12 nature of the solvent to be employed, provided that it has no described in detail by T. W. Greene et al., Protective Groups in adverse effect on the reaction orthereagents involved and that Organic Synthesis, 369-453, (1999), the disclosures of which it can dissolve reagents, at least to some extent. Examples of are incorporated herein by reference. The following exempli Suitable solvents include: halogenated hydrocarbons, such as dichloromethane, chloroform, carbon tetrachloride and 1,2- fies a typical reaction involving the protecting group tert dichloroethane; ethers, such as diethyl ether, diisopropyl butyldimethylsilyl. ether, tetrahydrofuran and dioxane; aromatic hydrocarbons, The deprotection of the hydroxyl groups is carried out with Such as benzene, toluene and nitrobenzene, amides, such as formamide, N,N-dimethylformamide, N,N-dimethylaceta an acid, such as acetic acid, hydrogen fluoride, hydrogen mide and hexamethylphosphoric triamide; amines, such as 10 fluoride-pyridine complex, or fluoride ion, such as tetrabuty N-methylmorpholine, triethylamine, tripropylamine, tributy lammonium fluoride (TBAF). lamine, diisopropylethylamine, dicyclohexylamine, N-meth The deprotection reaction is normally and preferably ylpiperidine, pyridine, 4-pyrrolidinopyridine, N,N-dimethy laniline and N,N-diethylaniline; alcohols, such as methanol, effected in the presence of solvent. There is no particular ethanol, propanol, 2-propanol and butanol; nitriles, such as 15 restriction on the nature of the solvent to be employed, pro acetonitrile and benzonitrile; sulfoxides, such as dimethyl vided that it has no adverse effect on the reaction or the Sulfoxide and Sulfolane, ketones, such as acetone and dieth reagents involved and that it can dissolve reagents, at least to ylketone; or mixed solvents thereof. Of these solvents, tet some extent. Examples of suitable solvents include, but are rahydrofuran, N,N-dimethylformamide or ethanol is pre ferred. not limited to: alcohol. Such as methanol, ethanol or mixed The reaction is carried out in the presence of a base. There solvents thereof. is likewise no particular restriction on the nature of the bases The deprotection reaction can take place over a wide range used, and any base commonly used in reactions of this type of temperatures, and the precise reaction temperature is not may equally be used here. Examples of Such bases include: critical to the invention. The preferred reaction temperature alkali metal hydroxides, such as lithium hydroxide, sodium 25 will depend upon Such factors as the nature of the solvent, and hydroxide and potassium hydroxide; alkali metal hydrides, the starting materials. However, in general, it is convenient to Such as lithium hydride, Sodium hydride and potassium carry out the reaction at a temperature of from about 0°C. to hydride; alkali metal alkoxides, such as Sodium methoxide, about 100° C. The time required for the reaction may also Sodium ethoxide and potassium tert-butoxide; alkali metal 30 vary widely, depending on many factors, notably the reaction carbonates, such as lithium carbonate, Sodium carbonate and potassium carbonate; alkali metal hydrogencarbonates, such temperature and the nature of the starting materials and sol as lithium hydrogencarbonate, sodium hydrogencarbonate vent employed. However, provided that the reaction is and potassium hydrogencarbonate; amines, such as N-meth effected under the preferred conditions outlined above, a ylmorpholine, triethylamine, tripropylamine, tributylamine, 35 period of from about 10 minutes to about 24 hours, will diisopropylethylamine, dicyclohexylamine, N-methylpiperi usually suffice. dine, pyridine, 4-pyrrolidinopyridine, picoline, 4-(N,N-dim ethylamino)pyridine, 2,6-di(tert-butyl)-4-methylpyridine, Method B quinoline, N,N-dimethylaniline, N,N-diethylaniline, 1,5-di 40 This illustrates the preparation of compounds of formula azabicyclo4.3.0non-5-ene (DBN), 1,4-diazabicyclo[2.2.2 (I). octane (DABCO) and 1,8-diazabicyclo5.4.0]undec-7-ene (DBU); alkali metal amides, such as lithium amide, sodium amide, potassium amide, lithium diisopropyl amide, potas Reaction Scheme B sium diisopropyl amide, Sodium diisopropyl amide, lithium 45 bis(trimethylsilyl)amide and potassium bis(trimethylsilyl)a- O R1a M mide. Of these, sodium hydride, potassium carbonate or RS N potassium tert-butoxide is preferred. X- R2a -- The reaction can take place over a wide range of tempera 50 N tures, and the precise reaction temperature is not critical to the invention. The preferred reaction temperature will depend XH upon Such factors as the nature of the solvent, and the starting materials. However, in general, it is convenient to carry out (IV) the reactionata temperature of from about 0°C. to about 120° 55 R9a C. The time required for the reaction may also vary widely, Lw depending on many factors, notably the reaction temperature A and the nature of the starting materials and solvent employed. B R5a Step B1 However, provided that the reaction is effected under the 60 preferred conditions outlined above, a period of from about 30 minutes to about 48 hours, will usually suffice. R8a R6a Deprotecting Step R7a In the case where R, R2, R,R,R,R, R7, R or 65 R" has a protected hydroxy group, the deprotection reaction will follow to yield a hydroxy group. This reaction is US 7,718,809 B2 14 (Step B2) -continued In this step, the compound (VI) is prepared by hydrolysis of the ester group of the compound of formula (V) with a base or O R" an acid. The reaction is normally and preferably effected in the RS N presence of solvent. There is no particular restriction on the X- R2a nature of the solvent to be employed, provided that it has no R9a N adverse effect on the reaction orthereagents involved and that it can dissolve reagents, at least to some extent. Examples of X Step B2 10 suitable solvents include: ethers, such as diethyl ether, diiso A HO propyl ether, tetrahydrofuran and dioxane; amides, such as formamide, N,N-dimethylformamide, N,N-dimethylaceta B R5a mide and hexamethylphosphoric triamide; alcohols, such as methanol, ethanol, propanol, 2-propanol and butanol; nitriles, Such as acetonitrile and benzonitrile; Sulfoxides, such as dim R8a R6a 15 ethyl sulfoxide and sulfolane; water; or mixed solvents thereof. Of these solvents, methanol, ethanol or tetrahydro R7a furan is preferred. (V) The reaction may be carried out in the presence of a base. O R1a There is likewise no particular restriction on the nature of the M bases used, and any base commonly used in reactions of this N type may equally be used here. Examples of Such bases HO include: alkali metal hydroxides, such as lithium hydroxide, A R2a Sodium hydroxide and potassium hydroxide; alkali metal car R9a N bonates, such as lithium carbonate, sodium carbonate and 25 potassium carbonate. Of these, lithium hydroxide or sodium X Step B3 hydroxide is preferred. A R3 R4 n NH1 The reaction may be carried out in the presence of an acid. B R5a There is likewise no particular restriction on the nature of the (VII) acids used, and any acid commonly used in reactions of this 30 type may equally be used here. Examples of Such acids include: carboxylic acids, such as acetic acid or propionic R8a R6a acid; acids, such as hydrochloric acid, Sulfuric acid or hydro bromic acid. Of these, hydrochloric acid is preferred. R7a The reaction can take place over a wide range of tempera (VI) tures, and the precise reaction temperature is not critical to the 35 invention. The preferred reaction temperature will depend O R1a upon Such factors as the nature of the solvent, and the starting 3 M materials. However, in general, it is convenient to carry out R N N the reactionata temperature of from about 0°C. to about 120° C. The time required for the reaction may also vary widely, R4 X- R2a 40 R9 N depending on many factors, notably the reaction temperature and the nature of the starting materials and solvent employed. X However, provided that the reaction is effected under the A preferred conditions outlined above, a period of from about 60 minutes to about 24 hours, will usually suffice. B R5 45 (Step B3) In this step, the compound (I) is prepared by amidation of R8 R6 the compound of formula (VI) with the compound of formula (VII), which is commercially available. If the compound of R7 formula (VI) or (VII) has hydroxy-protecting groups, the 50 deprotection reaction described in Method A will be applied (I) in an appropriate step. The reaction is normally and preferably effected in the In Reaction Scheme B. R is a C-C alkyl group or benzyl presence of solvent. There is no particular restriction on the nature of the solvent to be employed, provided that it has no group. 55 adverse effect on the reaction orthereagents involved and that (Step B1) it can dissolve reagents, at least to some extent. Examples of Suitable solvents include: halogenated hydrocarbons. Such as In this step, the compound of formula (V) is prepared by dichloromethane, chloroform, carbon tetrachloride and 1,2- nucleophilic substitution with LV of the compound of formula dichloroethane; aromatic hydrocarbons, such as benzene, (III) which is commercially available or may be prepared by 60 toluene and nitrobenzene, amides, such as formamide, N.N- the method described in WO 2000078751 or the following dimethylformamide, N,N-dimethylacetamide and hexameth ylphosphoric triamide; nitriles, such as acetonitrile and ben Method E to F, with the compound of formula (IV), which is Zonitrile; sulfoxides, such as dimethyl sulfoxide and commercially available or may be prepared by the methods sulfolane; or mixed solvents thereof. Of these, N,N-dimeth described in WO 2004.054984. The reaction may be carried 65 ylformamide is preferred. out under the same condition as described in Step A1 of The reaction is carried out in the presence of a base. There Method A. is likewise no particular restriction on the nature of the bases US 7,718,809 B2 15 used, and any base commonly used in reactions of this type may equally be used here. Examples of Such bases include: -continued amines, such as N-methylmorpholine, triethylamine, tripro R9a pylamine, tributylamine, diisopropylethylamine, dicyclo OH hexylamine, N-methylpiperidine, pyridine, 4-pyrrolidinopy A ridine, picoline, 4-(N,N-dimethylamino)pyridine, 2,6-di (tert-butyl)-4-methylpyridine, quinoline, N,N- B R5a Step C1 dimethylaniline, N,N-diethylaniline, DBN, DABCO and DBU. Of these, triethylamine or diisopropylethylamine is preferred. 10 R8a R6a The reaction is carried out in the presence of a condensing R7a agent. There is likewise no particular restriction on the nature of the condensing agents used, and any condensing agent (VIII) O RI commonly used in reactions of this type may equally be used 15 3 A here. Examples of Such condensing agents include: 2-halo-1- R NN N lower alkyl pyridinium halides, such as 2-chloro-1-methy 2 pyridinium iodide and 2-bromo-1-ethylpyridinium tetrafluo 4. 2 R roborate (BEP); diarylphosphorylazides, such as diphe R9 N nylphosphorylazide (DPPA); chloroformates, such as ethyl O chloroformate and isobutyl chloroformate; phosphorocyani A dates, such as diethyl phosphorocyanidate (DEPC); imida Zole derivatives, such as N,N'-carbonyldiimidazole (CDI): B R5 carbodiimide derivatives, such as N,N'-dicyclohexylcarbodi imide (DCC) and 1-(3-dimethylaminopropyl)-3-ethylcarbo 25 diimide hydrochloride (EDCI); iminium salts, such as 2-(1H R8 R6 benzotriazol-1-yl)-1,1,3,3-tetramethyluronium R7 hexafluorophosphate (HBTU) and tetramethyl fluoroforma midinium hexafluoro phosphate (TFFH); and phosphonium salts, such as benzotriazol-1-yloxytris(dimethylamino)phos 30 phonium hexafluorophosphate (BOP) and bromo-tris-pyrro lidino-phosphonium hexafluorophosphate (PyBrop). Of (Step C1) these, EDC1 or HBTU is preferred. In this step, the compound (Ia) is prepared by ether forma Reagents, such as 4-(N,N-dimethylamino)pyridine 35 tion reaction of the compound of formula (IIa), which is commercially available or may be prepared by the methods (DMAP), and 1-hydroxybenztriazole (HOBt), may be described in WO 2004.054984, with the compound (VIII), employed for this step. Of these, HOBt is preferred. which is commercially available or may be prepared by the The reaction can take place over a wide range of tempera method described in the following Method E to F. If the tures, and the precise reaction temperature is not critical to the compound of formula (IIa) or (VIII) has a hydroxy-protecting invention. The preferred reaction temperature will depend 40 group, the deprotection reaction described in Method A will upon Such factors as the nature of the solvent, and the starting be applied. materials. However, in general, it is convenient to carry out The reaction is normally and preferably effected in the the reaction at a temperature of from about 0°C. to about 80° presence of solvent. There is no particular restriction on the C. The time required for the reaction may also vary widely, 45 nature of the solvent to be employed, provided that it has no depending on many factors, notably the reaction temperature adverse effect on the reaction orthereagents involved and that and the nature of the starting materials and solvent employed. it can dissolve reagents, at least to some extent. Examples of However, provided that the reaction is effected under the Suitable solvents include: halogenated hydrocarbons. Such as preferred conditions outlined above, a period of from about dichloromethane, chloroform, carbon tetrachloride and 1,2- 30 minutes to about 48 hours, will usually suffice. 50 dichloroethane; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran and dioxane; aromatic hydrocarbons, Method C Such as benzene, toluene and nitrobenzene, amides, such as This illustrates the preparation of compounds of formula formamide, N,N-dimethylformamide, N,N-dimethylaceta (Ia) wherein X is oxygen atom. mide and hexamethylphosphoric triamide: nitriles, such as 55 acetonitrile and benzonitrile; or mixed solvents thereof. Of these, tetrahydrofuran or toluene is preferred. Reaction Scheme C The reaction is carried out in the presence of a condensing O agent. There is likewise no particular restriction on the nature R 3. 3 M of the condensing agents used, and any condensing agent R N N 60 commonly used in reactions of this type may equally be used here. Examples of Such condensing agents include: aZodicar R4a X-R- -- boxylic acid di-lower alkyl esters, such as diethylazodicar N boxylate (DEAD), diisopropylazodicarboxylate (DIAD) and di-tert-butyl azodicarboxylate (DTAD); azodicarboxamides, OH 65 such as N.N.N',N'-tetraisopropylazodicarboxamide (TIPA), (IIa) 1,1'-(azodicarbonyl)dipiperidine (ADDP) and N.N.N',N'-tet ramethylazodicarboxamide (TMAD); phosphoranes, such as US 7,718,809 B2 17 18 (cyanomethylene)tributylphosphorane (CMBP) and (cya (II), which is commercially available or may be prepared by nomethylene)trimethylphosphorane (CMMP). Of these, the methods described in WO 2004.054984, with the com DIAD is preferred. pound (IX), which is commercially available or can be pre Phosphine reagents. Such as triphenylphosphine, and tribu pared according to following Method G. After this reaction, tylphosphine, may be employed for this step. Of these, triph alkylation of hydroxy group (D1-b) with the compound of enylphosphine is preferred. formula (X) may follows. If the compound of formula (II) or The reaction can take place over a wide range of tempera (IX) has a hydroxy-protecting group, the deprotection reac tures, and the precise reaction temperature is not critical to the tion described in Method A will be applied. invention. The preferred reaction temperature will depend (D1-a) Epoxy Opening Reaction upon Such factors as the nature of the solvent, and the starting 10 The reaction is normally and preferably effected in the materials. However, in general, it is convenient to carry out presence of solvent. There is no particular restriction on the the reactionata temperature of from about 0°C. to about 120° nature of the solvent to be employed, provided that it has no C. The time required for the reaction may also vary widely, adverse effect on the reaction orthereagents involved and that depending on many factors, notably the reaction temperature it can dissolve reagents, at least to some extent. Examples of and the nature of the starting materials and solvent employed. 15 suitable solvents include: ethers, such as diethyl ether, diiso However, provided that the reaction is effected under the propyl ether, tetrahydrofuran and dioxane; amides, such as preferred conditions outlined above, a period of from about formamide, N,N-dimethylformamide, N,N-dimethylaceta 60 minutes to about 48 hours, will usually suffice. mide and hexamethylphosphoric triamide; alcohols, such as Method D methanol, ethanol, propanol, 2-propanol and butanol; nitriles, This illustrates the preparation of compounds of formula Such as acetonitrile and benzonitrile; Sulfoxides, such as dim (Ib) wherein R is a hydroxy group or a C-C alkoxy group. ethyl sulfoxide and Sulfolane, ketones, such as acetone and diethylketone; water; or mixed solvents thereof. Of these, ethanol in corporate with water is preferred. Reaction Scheme D 25 The reaction is carried out in the presence of a base. There O is likewise no particular restriction on the nature of the bases J. 3. used, and any base commonly used in reactions of this type R3 N N may equally be used here. Examples of Such bases include: alkali metal hydroxides, such as lithium hydroxide, sodium R4a X- R2a + hydroxide and potassium hydroxide; alkali metal carbonates, N Such as lithium carbonate, sodium carbonate and potassium carbonate; amines, such as N-methylmorpholine, triethy XH lamine, tripropylamine, tributylamine, diisopropylethy (II) lamine, dicyclohexylamine, N-methylpiperidine, pyridine, O 4-pyrrolidinopyridine, picoline, 4-(N,N-dimethylamino)py ridine, 2,6-di(tert-butyl)-4-methylpyridine, quinoline, N.N- dimethylaniline, N,N-diethylaniline, DBN, DABCO and B R5a Step D1 DBU. Of these, triethylamine is preferred. R-Hal The reaction can take place over a wide range of tempera tures, and the precise reaction temperature is not critical to the (X) 40 invention. The preferred reaction temperature will depend R8a R6a upon Such factors as the nature of the solvent, and the starting R7a materials. However, in general, it is convenient to carry out the reaction at a temperature of from about 20° C. to about (IX) 120° C. The time required for the reaction may also vary O 45 widely, depending on many factors, notably the reaction tem 3 J. perature and the nature of the starting materials and solvent R NN N employed. However, provided that the reaction is effected R4A 2 R2 under the preferred conditions outlined above, a period of R9b N 50 from about 60 minutes to about 48 hours, will usually suffice. X (D1-b) Alkylation A The reaction is normally and preferably effected in the presence of solvent. There is no particular restriction on the B R5 nature of the solvent to be employed, provided that it has no adverse effect on the reaction orthereagents involved and that it can dissolve reagents, at least to some extent. Examples of R8 R6 suitable solvents include: ethers, such as diethyl ether, diiso propyl ether, tetrahydrofuran and dioxane; aromatic hydro R7 carbons, such as benzene, toluene and nitrobenzene, amides, (Ib) 60 such as formamide, N,N-dimethylformamide, N,N-dimethy lacetamide and hexamethylphosphoric triamide; alcohols, In Reaction Scheme D. Halisahalogenatom, R is a C-C, Such as methanol, ethanol, propanol. 2-propanol and butanol: alkyl group and R’ is a hydroxy group or R O . nitriles, such as acetonitrile and benzonitrile; Sulfoxides, such as dimethyl sulfoxide and sulfolane; or mixed solvents (Step D1) 65 thereof. Of these, N,N-dimethylformamide is preferred. In this step, the compound of formula (Ib) is prepared by The reaction is carried out in the presence of a base. There epoxy opening reaction (D1-a) of the compound of formula is likewise no particular restriction on the nature of the bases US 7,718,809 B2 19 used, and any base commonly used in reactions of this type may equally be used here. Examples of Such bases include: -continued alkali metal hydroxides, such as lithium hydroxide, sodium OH Hal hydroxide and potassium hydroxide; alkali metal hydrides, B R5a B R5a Such as lithium hydride, Sodium hydride and potassium Step E4 hydride; alkali metal alkoxides, such as Sodium methoxide, He Sodium ethoxide and potassium tert-butoxide; alkali metal amides, such as lithium amide, Sodium amide, potassium R8a R6a R8a R6a amide, lithium diisopropyl amide, potassium diisopropyl 10 R7a R7a amide, Sodium diisopropylamide, lithium bis(trimethylsilyl) amide and potassium bis(trimethylsilyl)amide. Of these, (XVIa) (IIIa) sodium hydride is preferred. The reaction can take place over a wide range of tempera In Reaction Scheme E. Hal is a halogenatom and the same tures, and the precise reaction temperature is not critical to the 15 shall apply hereinafter. invention. The preferred reaction temperature will depend (Step E1) upon Such factors as the nature of the solvent, and the starting In this step, the compound of formula (XIV) is prepared by materials. However, in general, it is convenient to carry out Michael reaction (E1-a) of the compound of formula (XI) the reactionata temperature of from about 0°C. to about 100° with the compound of formula (XII) or by alkylation reaction C. The time required for the reaction may also vary widely, (E1-lb) of the compound of formula (XI) with the compound depending on many factors, notably the reaction temperature of formula (XIII). The compound of formula (XI), (XII) and and the nature of the starting materials and solvent employed. (XIII) are commercially available. However, provided that the reaction is effected under the (E1-a) Michael Reaction preferred conditions outlined above, a period of from about 25 The reaction is normally and preferably effected in the 30 minutes to about 48 hours, will usually suffice. presence or the absence of solvent. There is no particular restriction on the nature of the solvent to be employed, pro Method E vided that it has no adverse effect on the reaction or the This illustrates the preparation of compounds of formula reagents involved and that it can dissolve reagents, at least to (IIIa) wherein A is CH. 30 some extent. Examples of suitable solvents include: ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran and dioxane: amides, such as formamide, N,N-dimethylforma Reaction Scheme E mide, N,N-dimethylacetamide and hexamethylphosphoric HB R5a triamide; alcohols, such as methanol, ethanol, propanol, 35 2-propanol and butanol; nitriles, such as acetonitrile and ben Step E1 Zonitrile; sulfoxides, such as dimethyl sulfoxide and sul folane; or mixed solvents thereof. Of these, the reaction in the R8a R6a a) 21 NoN absence of solvent is preferred. R7a (XII) The reaction is carried out in the presence of a base. There 40 is likewise no particular restriction on the nature of the bases (XI) O used, and any base commonly used in reactions of this type may equally be used here. Examples of Such bases include: b) Hal --- OH alkali metal hydroxides, such as lithium hydroxide, sodium (XIII) hydroxide and potassium hydroxide; alkali metal hydrides, HO O 45 Such as lithium hydride, Sodium hydride and potassium hydride; alkali metal alkoxides, such as Sodium methoxide, Sodium ethoxide and potassium tert-butoxide; alkali metal carbonates, such as lithium carbonate, sodium carbonate and potassium carbonate; amines, such as N-methylmorpholine, B Sa Step E2 He triethylamine, tripropylamine, tributylamine, diisopropyl ethylamine, dicyclohexylamine, N-methylpiperidine, pyri dine, 4-pyrrolidinopyridine, picoline, 4-(N,N-dimethy lamino)pyridine, 2,6-di(tert-butyl)-4-methylpyridine, R8a R6a quinoline, N,N-dimethylaniline, N,N-diethylaniline, DBN, R7a DABCO, DBU and benzyltrimethylammonium hydroxide: (XIV) alkali metal amides, such as lithium amide, sodium amide, O potassium amide, lithium diisopropyl amide, potassium diisopropyl amide, sodium diisopropyl amide, lithium bis (trimethylsilyl)amide and potassium bis(trimethylsilyl) B R5a Step E3 60 amide. Of these, benzyltrimethylammonium hydroxide or Her sodium methoxide is preferred. The reaction can take place over a wide range of tempera R8a R6a tures, and the precise reaction temperature is not critical to the R7a invention. The preferred reaction temperature will depend 65 upon Such factors as the nature of the solvent, and the starting (XVa) materials. However, in general, it is convenient to carry out the reaction at a temperature of from about 20° C. to about US 7,718,809 B2 21 22 120° C. The time required for the reaction may also vary Introduction of the Hydroxy-Protecting Group widely, depending on many factors, notably the reaction tem In the case of the compound of formula (IIIa) having a perature and the nature of the starting materials and solvent hydroxy group, if necessary, the reaction may be accom employed. However, provided that the reaction is effected plished by protecting the hydroxy group. under the preferred conditions outlined above, a period of 5 The introduction of the hydroxy-protecting group can be from about 60 minutes to about 48 hours, will usually suffice. carried out at an appropriate step before the reaction affected After the above procedure, hydrolysis is carried out by by the hydroxy group. adding an acid in a solvent to produce the compound of This reaction is described in detail by T. W. Greene et al., formula (XIV), and may be carried out in a usual hydrolysis Protective Groups in Organic Synthesis, 369-453, (1999), the condition. The acid may include, for example, inorganic acids 10 disclosures of which are incorporated herein by reference. Such as hydrochloric acid, hydrobromic acid and Sulfuric The following exemplifies a typical reaction involving the acid. It is preferably hydrochloric acid. The solvent may protecting group tert-butyldimethylsilyl. include, for example, water, alcohols such as methanol, etha For example, when the hydroxy-protecting group is a “tert nol, propanol and tert-butanol; ethers such as diethyl ether, butyldimethylsilyl', this step is conducted by reacting with a dimethoxyethane, tetrahydrofuran, diethoxymethane and 15 desired hydroxy-protecting group halide in an inert Solvent in dioxane; or mixed solvents thereof. It is preferably water. The the presence of a base. reaction temperature varies depending on the starting com Examples of suitable solvents include: halogenated hydro pound, the reagent and the solvent, however, it is usually from carbons, such as dichloromethane, chloroform, carbon tetra 20° C. to the reflux temperature. The reaction time varies chloride and 1,2-dichloroethane; ethers, such as diethyl ether, depending on the starting compound, the reagent, the solvent diisopropyl ether, tetrahydrofuran and dioxane; aromatic and the reaction temperature, however, it is usually from 60 hydrocarbons, such as benzene, toluene and nitrobenzene: minutes to 24 hours. amides, such as formamide, N,N-dimethylformamide, N.N- dimethylacetamide and hexamethylphosphoric triamide; or (E1-lb) Alkylation Reaction mixed solvents thereof. Of these, tetrahydrofuran or N.N- The reaction is normally and preferably effected in the 25 dimethylformamide is preferred. presence of solvent. There is no particular restriction on the Examples of the hydroxy-protecting group halide usable in nature of the solvent to be employed, provided that it has no the above reaction include trimethylsilyl chloride, triethylsi adverse effect on the reaction orthereagents involved and that lyl chloride, tert-butyldimethylsilyl chloride, tert-butyldim it can dissolve reagents, at least to some extent. Examples of ethylsilyl bromide, acetyl chloride are preferred. suitable solvents include: ethers, such as diethyl ether, diiso 30 Examples of the base include alkali metal hydroxides such propyl ether, tetrahydrofuran and dioxane; aromatic hydro as lithium hydroxide, sodium hydroxide and potassium carbons, such as benzene, toluene and nitrobenzene: amides, hydroxide, alkali metal carbonates such as lithium carbonate, such as formamide, N,N-dimethylformamide, N,N-dimethy Sodium carbonate and potassium carbonate, and organic lacetamide and hexamethylphosphoric triamide; alcohols, amines such as triethylamine, tributylamine, N-methylmor Such as methanol, ethanol, propanol. 2-propanol and butanol: 35 pholine, pyridine, imidazole, 4-dimethylaminopyridine, nitriles, such as acetonitrile and benzonitrile; Sulfoxides, such picoline, lutidine, collidine, DBN and DBU. Out of these, as dimethylsulfoxide and Sulfolane; ketones, such as acetone triethylamine, imidazole, or pyridine is preferred. Upon use and diethylketone; water; or mixed solvents thereof. Of these, of an organic amine in the liquid form, it also serves as a water is preferred. Solvent when used in large excess. The reaction is carried out in the presence of a base. There 40 Although the reaction temperature differs with the nature is likewise no particular restriction on the nature of the bases of the starting compound, the halide and the solvent, it usually used, and any base commonly used in reactions of this type ranges from 0°C. to 80° C. (preferably 0 to 30° C.). Although may equally be used here. Examples of Such bases include: the reaction time differs with the reaction temperature or the alkali metal hydroxides, such as lithium hydroxide, sodium like, it ranges from 10 minutes to 2 days (preferably 30 hydroxide and potassium hydroxide; alkali metal hydrides, 45 minutes to 1 day). Such as lithium hydride, Sodium hydride and potassium (Step E2) hydride; alkali metal alkoxides, such as Sodium methoxide, In this step, the compound of formula (XVa) is prepared by Sodium ethoxide and potassium tert-butoxide; alkali metal Friedel Crafts reaction (E2-a) after halogenation (E2-b) or by carbonates, such as lithium carbonate, Sodium carbonate and cyclization (E2-c) of the compound of formula (XIV). potassium carbonate; alkali metal amides. Such as lithium 50 amide, Sodium amide, potassium amide, lithium diisopropyl (E2-a) Friedel Crafts Reaction amide, potassium diisopropyl amide, sodium diisopropyl The reaction is normally and preferably effected in the amide, lithium bis(trimethylsilyl)amide and potassium bis presence or the absence of solvent. There is no particular (trimethylsilyl)amide. Of these, sodium hydroxide is pre restriction on the nature of the solvent to be employed, pro ferred. 55 vided that it has no adverse effect on the reaction or the The reaction can take place over a wide range of tempera reagents involved and that it can dissolve reagents, at least to tures, and the precise reaction temperature is not critical to the Some extent. Examples of Suitable solvents include: haloge invention. The preferred reaction temperature will depend nated hydrocarbons, such as dichloromethane, chloroform, upon Such factors as the nature of the solvent, and the starting carbon tetrachloride, 1.1.2.2-tetrachloroehane and 1.2- materials. However, in general, it is convenient to carry out 60 dichloroethane; aromatic hydrocarbons, such as benzene, the reaction at a temperature of from about 20° C. to about toluene and nitrobenzene: carbon disulfide; or mixed solvents 100° C. The time required for the reaction may also vary thereof. Of these, dichloromethane or carbon disulfide is pre widely, depending on many factors, notably the reaction tem ferred. perature and the nature of the starting materials and solvent The reaction is carried out in the presence of an acid. There employed. However, provided that the reaction is effected 65 is likewise no particular restriction on the nature of the acids under the preferred conditions outlined above, a period of used, and any acid commonly used in reactions of this type from about 60 minutes to about 24 hours, will usually suffice. may equally be used here. Examples of such acids include: US 7,718,809 B2 23 24 Lewis acids, such as BFs. AlCl, AlBrs. FeCls, AgCl, ZnI, used, and any acid commonly used in reactions of this type ZnCl2, Fe(NO), CFSO Si (CH), Yb(CFSO), and may equally be used here. Examples of Such acids include: SnCla. Of these, AlCl is preferred. acids, Such as hydrochloric acid, Sulfuric acid, or hydrobro The reaction can take place over a wide range of tempera mic acid; acids, such as trifluoro acetic acid, or polyphospho tures, and the precise reaction temperature is not critical to the 5 ric acid. Of these, polyphosphoric acid is preferred. invention. The preferred reaction temperature will depend The reaction can take place over a wide range of tempera upon Such factors as the nature of the solvent, and the starting tures, and the precise reaction temperature is not critical to the materials. However, in general, it is convenient to carry out invention. The preferred reaction temperature will depend the reactionata temperature of from about 0°C. to about 150° upon Such factors as the nature of the solvent, and the starting C. The time required for the reaction may also vary widely, 10 materials. However, in general, it is convenient to carry out depending on many factors, notably the reaction temperature the reaction at a temperature of from about 20° C. to about and the nature of the starting materials and solvent employed. 150° C. The time required for the reaction may also vary However, provided that the reaction is effected under the widely, depending on many factors, notably the reaction tem preferred conditions outlined above, a period of from about perature and the nature of the starting materials and solvent 30 minutes to about 24 hours, will usually suffice. 15 employed. However, provided that the reaction is effected (E2-b) Halogenation under the preferred conditions outlined above, a period of The reaction is normally and preferably effected in the from about 30 minutes to about 24 hours, will usually suffice. presence of solvent. There is no particular restriction on the (Step E3) nature of the solvent to be employed, provided that it has no In this step, the compound (XVIa) is prepared by reduction adverse effect on the reaction orthereagents involved and that of the carbonyl group of the compound of formula (XVa). it can dissolve reagents, at least to some extent. Examples of The reaction is normally and preferably effected in the Suitable solvents include: halogenated hydrocarbons, such as presence of solvent. There is no particular restriction on the dichloromethane, chloroform, carbon tetrachloride and 1,2- nature of the solvent to be employed, provided that it has no dichloroethane; ethers, such as diethyl ether, diisopropyl adverse effect on the reaction orthereagents involved and that ether, tetrahydrofuran and dioxane; amides, such as forma 25 it can dissolve reagents, at least to some extent. Examples of mide, N,N-dimethylformamide, N,N-dimethylacetamide and Suitable solvents include: halogenated hydrocarbons. Such as hexamethylphosphoric triamide; amines, such as nitriles, dichloromethane, chloroform, carbon tetrachloride and 1,2- Such as acetonitrile and benzonitrile; or mixed solvents dichloroethane; ethers, such as diethyl ether, diisopropyl thereof. Of these, 1,2-dichloroethane or dichloromethane is ether, tetrahydrofuran and dioxane; aromatic hydrocarbons, preferred. 30 Such as benzene, toluene and nitrobenzene: Sulfoxides. Such The reaction is carried out in the presence of a halogenating as dimethylsulfoxide and sulfolane, alcohols, such as metha agent. There is likewise no particular restriction on the nature nol, ethanol, propanol, 2-propanol and butanol; or mixed of the halogenating agents used, and any halogenating agent solvents thereof. Of these, methanol or tetrahydrofuran is commonly used in reactions of this type may equally be used preferred. here. Examples of such halogenating agents include: thionyl 35 The reaction is carried out in the presence of a reducing chloride, oxalyl chloride and phosphorus oxychloride. Of agent. There is likewise no particular restriction on the nature these, thionyl chloride is preferred. of the reducing agents used, and any reducing agent com The reaction can take place over a wide range of tempera monly used in reactions of this type may equally be used here. tures, and the precise reaction temperature is not critical to the Examples of Such reducing agents include: metal borohy invention. The preferred reaction temperature will depend 40 drides, such as sodium borohydride, lithium borohydride and upon Such factors as the nature of the solvent, and the starting Sodium cyanoborohydride; hydride compounds, such as materials. However, in general, it is convenient to carry out lithium aluminum hydride and diisobutyl aluminum hydride. the reaction at a temperature of from about 0°C. to about 80° Of these, sodium borohydride is preferred. C. The time required for the reaction may also vary widely, The reaction can take place over a wide range of tempera depending on many factors, notably the reaction temperature 45 tures, and the precise reaction temperature is not critical to the and the nature of the starting materials and solvent employed. invention. The preferred reaction temperature will depend However, provided that the reaction is effected under the upon Such factors as the nature of the solvent, and the starting preferred conditions outlined above, a period of from about materials. However, in general, it is convenient to carry out 10 minutes to about 8 hours will usually suffice. the reaction at a temperature of from about 0°C. to about 80° 50 C. The time required for the reaction may also vary widely, (E2-c) Cyclization depending on many factors, notably the reaction temperature The reaction is normally and preferably effected in the and the nature of the starting materials and solvent employed. presence or absence of solvent. There is no particular restric However, provided that the reaction is effected under the tion on the nature of the solvent to be employed, provided that preferred conditions outlined above, a period of from about it has no adverse effect on the reaction or the reagents 55 involved and that it can dissolve reagents, at least to some 10 minutes to about 8 hours will usually suffice. extent. Examples of Suitable solvents include: halogenated (Step E4) hydrocarbons, such as dichloromethane, chloroform, carbon In this step, the compound of formula (IIIa) is prepared by tetrachloride and 1,2-dichloroethane; ethers, such as diethyl halogenation of the hydroxy group of the compound of for ether, diisopropyl ether, tetrahydrofuran and dioxane; aro 60 mula (XVIa). matic hydrocarbons. Such as benzene, toluene and nitroben The reaction is normally and preferably effected in the Zene; amides, such as formamide, N,N-dimethylformamide, presence or the absence of solvent. There is no particular N,N-dimethylacetamide and hexamethylphosphoric tria restriction on the nature of the solvent to be employed, pro mide; or mixed solvents thereof. Of these, dichloromethane vided that it has no adverse effect on the reaction or the or the absence of solvent is preferred. 65 reagents involved and that it can dissolve reagents, at least to The reaction is carried out in the presence of an acid. There Some extent. Examples of Suitable solvents include: haloge is likewise no particular restriction on the nature of the acids nated hydrocarbons, such as dichloromethane, chloroform, US 7,718,809 B2 25 26 carbon tetrachloride and 1,2-dichloroethane; ethers, such as The reaction is carried out in the presence of a halogenating diethyl ether, diisopropyl ether, tetrahydrofuran and dioxane: agent. There is likewise no particular restriction on the nature aromatic hydrocarbons, such as benzene, toluene and of the halogenating agents used, and any halogenating agent nitrobenzene: amides, such as formamide, N,N-dimethylfor commonly used in reactions of this type may equally be used mamide, N,N-dimethylacetamide and hexamethylphospho here. Examples of such halogenating agents include: thionyl ric triamide; amines, such as N-methylmorpholine, triethy chloride, oxalyl chloride, phosphorus pentachloride and lamine, tripropylamine, tributylamine, phosphorus oxychloride. Of these, thionyl chloride is pre diisopropylethylamine, dicyclohexylamine, N-methylpiperi ferred. dine, pyridine, 4-pyrrolidinopyridine, N,N-dimethylaniline The reaction can take place over a wide range of tempera and N,N-diethylaniline; nitriles, such as acetonitrile and ben 10 tures, and the precise reaction temperature is not critical to the Zonitrile; sulfoxides, such as dimethyl sulfoxide and sul invention. The preferred reaction temperature will depend folane; or mixed solvents thereof. Of these, diethyl ether or upon Such factors as the nature of the solvent, and the starting tetrahydrofuran is preferred. materials. However, in general, it is convenient to carry out The reaction may be carried out in the presence of a base. the reactionata temperature of from about 0°C. to about 100° There is likewise no particular restriction on the nature of the 15 C. The time required for the reaction may also vary widely, bases used, and any base commonly used in reactions of this depending on many factors, notably the reaction temperature type may equally be used here. Examples of Such bases and the nature of the starting materials and solvent employed. include: amines. Such as N-methylmorpholine, triethylamine, However, provided that the reaction is effected under the tripropylamine, tributylamine, diisopropylethylamine, dicy preferred conditions outlined above, a period of from about clohexylamine, N-methylpiperidine, pyridine, 4-pyrrolidi 20 10 minutes to about 8 hours will usually suffice. nopyridine, picoline, 4-(N,N-dimethylamino)pyridine, 2,6-di (tert-butyl)-4-methylpyridine, quinoline, N,N- Method F dimethylaniline, N,N-diethylaniline, DBN, DABCO and This illustrates the preparation of compounds of formula DBU. Of these, pyridine is preferred. (IIIb) wherein B is CH.

Reaction Scheme F CORC Hal CORC CORC H3C R5a R5a 1N R5a Step F1 Step F2 ROC A. Step F3 ------O R8a R6a R8a R6a --- AH R8a R6a R7a R7a Rdo R7a (XVII) (XVIII) (XIX) (XX) CORd O O OH Hal A. A A A

R5a --Step F4 R5a ---Step F5 R5a --Step F6 R5a

R8a R6a R8a R6a R8a R6a R8a R6a R7a R7a R7a R7a (XXI) (XVb) (XVIb) (IIIb) Step p

Step r US 7,718,809 B2 27 28

-continued

Hal R5a R5a Step F7 Roc1NA Hs O 8a 6a 8a. 6a R R --- AH R R R7a Rdo R7a (XXII) (XXIII) (XXIV)

In Reaction Scheme F. R and Rindependently represent (Step F2) a C-C alkyl group. In this step, the compound of formula (XX) is prepared by ether formation reaction of the compound of formula (XVIII) (Step F1) with the compound of formula (XIX), which is commercially In this step, the compound of formula (XVIII) is prepared available. by halogenation of the methyl group of the compound of 25 The reaction is normally and preferably effected in the formula (XVII). presence of solvent. There is no particular restriction on the The reaction is normally and preferably effected in the nature of the solvent to be employed, provided that it has no presence of solvent. There is no particular restriction on the adverse effect on the reaction orthereagents involved and that nature of the solvent to be employed, provided that it has no 30 it can dissolve reagents, at least to some extent. Examples of adverse effect on the reaction orthereagents involved and that suitable solvents include: ethers, such as diethyl ether, diiso it can dissolve reagents, at least to some extent. Examples of propyl ether, tetrahydrofuran and dioxane: aromatic hydro Suitable solvents include: halogenated hydrocarbons, such as carbons, such as benzene, toluene and nitrobenzene, amides, dichloromethane, chloroform, carbon tetrachloride and 1,2- such as formamide, N,N-dimethylformamide, N,N-dimethy dichloroethane; ethers, such as diethyl ether, diisopropyl 35 lacetamide and hexamethylphosphoric triamide; nitriles, ether, tetrahydrofuran and dioxane, nitriles, such as acetoni Such as acetonitrile and benzonitrile; Sulfoxides, such as dim trile and benzonitrile; sulfoxides, such as dimethyl sulfoxide ethyl sulfoxide and sulfolane; or mixed solvents thereof. Of and sulfolane; or mixed solvents thereof. Of these, carbon these, N,N-dimethylformamide or tetrahydrofuran is pre tetrachloride or 1,2-dichloroethane is preferred. 40 ferred. The reaction is carried out in the presence of a halogenating The reaction is carried out in the presence of a base. There agent. There is likewise no particular restriction on the nature is likewise no particular restriction on the nature of the bases of the halogenating agents used, and any halogenating agent used, and any base commonly used in reactions of this type 45 may equally be used here. Examples of Such bases include: commonly used in reactions of this type may equally be used alkali metal hydroxides, such as lithium hydroxide, sodium here. Examples of such halogenating agents include: Succin hydroxide and potassium hydroxide; alkali metal hydrides, imides, such as N-bromosuccinimide (NBS), N-chlorosuc Such as lithium hydride, Sodium hydride and potassium cinimide (NCS); bromine. Of these, NBS is preferred. hydride; alkali metal alkoxides, such as Sodium methoxide, Reagents, such as benzoyl peroxide and 2,2'-aZobis(isobu 50 Sodium ethoxide and potassium tert-butoxide; alkali metal tyronitrile) (AlBN) may be employed for this step. Of these, amides, such as lithium amide, Sodium amide, potassium benzoyl peroxide is preferred. amide, lithium diisopropyl amide, potassium diisopropyl amide, sodium diisopropylamide, lithium bis(trimethylsilyl) The reaction can take place over a wide range of tempera 55 amide and potassium bis(trimethylsilyl)amide. Of these, tures, and the precise reaction temperature is not critical to the sodium hydride is preferred. invention. The preferred reaction temperature will depend The reaction can take place over a wide range of tempera upon Such factors as the nature of the solvent, and the starting tures, and the precise reaction temperature is not critical to the materials. However, in general, it is convenient to carry out invention. The preferred reaction temperature will depend the reactionata temperature of from about 0°C. to about 100° 60 upon Such factors as the nature of the solvent, and the starting C. The time required for the reaction may also vary widely, materials. However, in general, it is convenient to carry out depending on many factors, notably the reaction temperature the reaction at a temperature of from about 20° C. to about and the nature of the starting materials and solvent employed. 150° C. The time required for the reaction may also vary However, provided that the reaction is effected under the 65 widely, depending on many factors, notably the reaction tem preferred conditions outlined above, a period of from about perature and the nature of the starting materials and solvent 30 minutes to about 24 hours will usually suffice. employed. However, provided that the reaction is effected US 7,718,809 B2 29 30 under the preferred conditions outlined above, a period of include: alkali metal hydroxides, such as lithium hydroxide, from about 60 minutes to about 48 hours, will usually suffice. Sodium hydroxide and potassium hydroxide; alkali metal car (Step F3) bonates, such as lithium carbonate, sodium carbonate and potassium carbonate. Of these, sodium hydroxide is pre In this step, the compound of formula (XXI) is prepared by 5 cyclization (Dieckmann Condensation) of the compound of ferred. formula (XX). The reaction may be carried out in the presence of an acid. The reaction is normally and preferably effected in the There is likewise no particular restriction on the nature of the presence of solvent. There is no particular restriction on the acids used, and any acid commonly used in reactions of this nature of the solvent to be employed, provided that it has no 10 type may equally be used here. Examples of Such acids adverse effect on the reaction orthereagents involved and that include: carboxylic acids, such as acetic acid or propionic it can dissolve reagents, at least to some extent. Examples of acid; acids, such as hydrochloric acid, Sulfuric acid, or hydro suitable solvents include: ethers, such as diethyl ether, diiso bromic acid. Of these, hydrochloric acid is preferred. propyl ether, tetrahydrofuran and dioxane; aromatic hydro 15 The reaction can take place over a wide range of tempera carbons, such as benzene, toluene and nitrobenzene: alco tures, and the precise reaction temperature is not critical to the hols, such as methanol, ethanol, propanol, 2-propanol and invention. The preferred reaction temperature will depend butanol; or mixed solvents thereof. Of these, toluene is pre upon Such factors as the nature of the solvent, and the starting ferred. materials. However, in general, it is convenient to carry out the reaction at a temperature of from about 20° C. to about The reaction is carried out in the presence of a base. There 120° C. The time required for the reaction may also vary is likewise no particular restriction on the nature of the bases widely, depending on many factors, notably the reaction tem used, and any base commonly used in reactions of this type perature and the nature of the starting materials and solvent may equally be used here. Examples of Such bases include: employed. However, provided that the reaction is effected alkali metal. Such as lithium and sodium; alkali metal 25 under the preferred conditions outlined above, a period of hydrides, such as lithium hydride, Sodium hydride and potas from about 60 minutes to about 48 hours, will usually suffice. sium hydride; alkali metal amides, such as lithium amide, Sodium amide, potassium amide, lithium diisopropyl amide, (Step F5) potassium diisopropyl amide, Sodium diisopropyl amide, In this step, the compound of formula (XVIb) is prepared lithium bis(trimethylsilyl)amide and potassium bis(trimeth 30 by reduction of the compound of formula (XVb). The reac ylsilyl)amide. Of these, sodium is preferred. tion may be carried out under the same condition as described The reaction can take place over a wide range of tempera in Step E3 of Method E. tures, and the precise reaction temperature is not critical to the invention. The preferred reaction temperature will depend (Step F6) 35 In this step, the compound of formula (IIIb) is prepared by upon Such factors as the nature of the solvent, and the starting halogenation of the compound of formula (XVIb). The reac materials. However, in general, it is convenient to carry out tion may be carried out under the same condition as described the reactionata temperature of from about 0°C. to about 150° in Step E4 of Method E. If the compound of formula (IIIb) has C. The time required for the reaction may also vary widely, hydroxy groups, the reaction for introducing the hydroxy depending on many factors, notably the reaction temperature 40 protecting group described in Method E will be applied in an and the nature of the starting materials and solvent employed. appropriate step. However, provided that the reaction is effected under the preferred conditions outlined above, a period of from about (Step F7) 30 minutes to about 24 hours, will usually suffice. In this step, the compound of formula (XXIV) is prepared 45 by ether formation reaction of the compound of formula (Step F4) (XXII) with the compound of formula (XXIII), which is In this step, the compound of formula (XVb) is prepared by commercially available. The reaction may be carried out decarboxylation of the compound of formula (XXI). under the same condition as described in Step F2 of Method The reaction is normally and preferably effected in the F. 50 presence of solvent. There is no particular restriction on the (Step F8) nature of the solvent to be employed, provided that it has no In this step, the compound of formula (XXV) is prepared adverse effect on the reaction orthereagents involved and that by hydrolysis of the compound of formula (XXIV). The reac it can dissolve reagents, at least to some extent. Examples of tion may be carried out under the same condition as described suitable solvents include: ethers, such as diethyl ether, diiso 55 in Step B2 of Method B. propyl ether, tetrahydrofuran and dioxane, amides, such as formamide, N,N-dimethylformamide, N,N-dimethylaceta (Step F9) mide and hexamethylphosphoric triamide; alcohols, such as In this step, the compound of formula (XVb) is prepared by methanol, ethanol, propanol. 2-propanol, ethylene glycol and cyclization (F9-a) of the compound of formula (XXIV) or by 60 formation of acid halide (F9-b) followed by Friedel Crafts butanol; nitriles, such as acetonitrile and benzonitrile; sulfox reaction (F9-c) of the compound of formula (XXV). The ides, such as dimethyl Sulfoxide and Sulfolane; water, or reaction may be carried out under the same condition as mixed solvents thereof. Of these, ethanol is preferred. described in Step E2 of Method E. The reaction may be carried out in the presence of a base. Method G There is likewise no particular restriction on the nature of the 65 bases used, and any base commonly used in reactions of this This illustrates the preparation of compounds of formula type may equally be used here. Examples of Such bases (IX). US 7,718,809 B2

Reaction Scheme G Step G2

Hal O OH OH A. A. N A. A B R5a R5a B R5a B R5a Step G1 Step G3 Step G4 -- -3- --- HO R8a R6a R8a R6a R8a R6a R8a R6a R7a R7a R7a R7a (XXVI) (XXVII) (XXVIII) (IX)

(Step G1) nature of the solvent to be employed, provided that it has no In this step, the compound of formula (XXVII) is prepared adverse effect on the reaction orthereagents involved and that by intermolecular dehydration of compound of formula it can dissolve reagents, at least to some extent. Examples of (XXVI), which is commercially available or may be prepared 25 Suitable solvents include: halogenated hydrocarbons. Such as by the foregoing method E or F. dichloromethane, chloroform, carbon tetrachloride and 1,2- The reaction is normally and preferably effected in the dichloroethane; ethers, such as diethyl ether, diisopropyl presence of solvent. There is no particular restriction on the ether, tetrahydrofuran and dioxane; aromatic hydrocarbons, nature of the solvent to be employed, provided that it has no Such as benzene, toluene and nitrobenzene, amides, such as adverse effect on the reaction orthereagents involved and that 30 formamide, N,N-dimethylformamide, N,N-dimethylaceta it can dissolve reagents, at least to some extent. Examples of mide and hexamethylphosphoric triamide: nitriles, such as suitable solvents include: ethers, such as diethyl ether, diiso acetonitrile and benzonitrile; sulfoxides, such as dimethyl propyl ether, tetrahydrofuran and dioxane; aromatic hydro Sulfoxide and Sulfolane; ketones, such as acetone and dieth carbons, such as benzene, toluene and nitrobenzene, amides, ylketone; water; or mixed solvents thereof. Of these, dichlo such as formamide, N,N-dimethylformamide, N,N-dimethy 35 romethane is preferred. lacetamide and hexamethylphosphoric triamide; nitriles, The reaction is carried out in the presence of a base. There Such as acetonitrile and benzonitrile; Sulfoxides, such as dim is likewise no particular restriction on the nature of the bases ethyl sulfoxide and sulfolane; or mixed solvents thereof. Of used, and any base commonly used in reactions of this type these, toluene is preferred. may equally be used here. Examples of Such bases include: The reaction is carried out in the presence of an acid. There 40 alkali metal hydroxides, such as lithium hydroxide, sodium is likewise no particular restriction on the nature of the acids hydroxide and potassium hydroxide; alkali metal carbonates, used, and any acid commonly used in reactions of this type Such as lithium carbonate, sodium carbonate and potassium may equally be used here. Examples of Such acids include: carbonate; alkali metal hydrogencarbonates, such as lithium carboxylic acids, such as acetic acid, trifluoro acetic acid or hydrogencarbonate, Sodium hydrogencarbonate and potas propionic acid; acids, such as hydrochloric acid, hydrochloric 45 sium hydrogencarbonate; amines, such as N-methylmorpho acid, Sulfuric acid, or p-toluenesulfonic acid; Lewis acids, line, triethylamine, tripropylamine, tributylamine, diisopro such as BF, AlCls. FeCl, AgCl, ZnI, Fe(NO), CFSOSi pylethylamine, dicyclohexylamine, N-methylpiperidine, (CH), Yb(CFSO) and SnCla. Of these, p-toluenesulfonic pyridine, 4-pyrrolidinopyridine, picoline, 4-(N,N-dimethy acid is preferred. lamino)pyridine, 2,6-di(tert-butyl)-4-methylpyridine, quino The reaction can take place over a wide range of tempera 50 line, N,N-dimethylaniline, N,N-diethylaniline, DBN, tures, and the precise reaction temperature is not critical to the DABCO and DBU. Of these, sodium hydrogencarbonate is invention. The preferred reaction temperature will depend preferred. upon Such factors as the nature of the solvent, and the starting materials. However, in general, it is convenient to carry out The reaction is carried out in the presence of an oxidizing the reactionata temperature of from about 0°C. to about 150° 55 agent. There is likewise no particular restriction on the nature C. The time required for the reaction may also vary widely, of the oxidizing agents used, and any oxidizing agent com depending on many factors, notably the reaction temperature monly used in reactions of this type may equally be used here. and the nature of the starting materials and solvent employed. Examples of Such oxidizing agents include: peroxy acids, However, provided that the reaction is effected under the such as 3-chloroperbenzoic acid (MCPBA), perbenzoic acid, 60 peracetic acid and trifluoroperacetic acid; peroxides. Such as preferred conditions outlined above, a period of from about hydrogen peroxide and tert-butyl hydroperoxide. Of these, 10 minutes to about 24 hours, will usually suffice. MCPBA is preferred. (Step G2) The reaction can take place over a wide range of tempera In this step, the compound of formula (IX) is prepared by tures, and the precise reaction temperature is not critical to the epoxydation of the compound of formula (XXVII). 65 invention. The preferred reaction temperature will depend The reaction is normally and preferably effected in the upon Such factors as the nature of the solvent, and the starting presence of solvent. There is no particular restriction on the materials. However, in general, it is convenient to carry out US 7,718,809 B2 33 34 the reaction at a temperature of from about -78°C. to about acetonitrile and benzonitrile; sulfoxides, such as dimethyl 100° C. The time required for the reaction may also vary sulfoxide and sulfolane; or mixed solvents thereof. Of these, widely, depending on many factors, notably the reaction tem diethyl ether is preferred. perature and the nature of the starting materials and solvent The reaction is carried out in the presence of a base. There employed. However, provided that the reaction is effected 5 is likewise no particular restriction on the nature of the bases under the preferred conditions outlined above, a period of used, and any base commonly used in reactions of this type from about 10 minutes to about 24 hours, will usually suffice. may equally be used here. Examples of Such bases include: (Step G3) alkali metal hydroxides, such as lithium hydroxide, sodium In this step, the compound of formula (XXVIII) is prepared hydroxide and potassium hydroxide; alkali metal carbonates, 10 Such as lithium carbonate, sodium carbonate and potassium by electrophilic addition of the compound of formula (XX carbonate; alkali metal hydrogencarbonates, such as lithium VII). hydrogencarbonate, Sodium hydrogencarbonate and potas The reaction is normally and preferably effected in the sium hydrogencarbonate; amines, such as N-methylmorpho presence of solvent. There is no particular restriction on the line, triethylamine, tripropylamine, tributylamine, diisopro nature of the solvent to be employed, provided that it has no 15 pylethylamine, dicyclohexylamine, N-methylpiperidine, adverse effect on the reaction orthereagents involved and that pyridine, 4-pyrrolidinopyridine, picoline, 4-(N,N-dimethy it can dissolve reagents, at least to some extent. Examples of Suitable solvents include: halogenated hydrocarbons, such as lamino)pyridine, 2,6-di(tert-butyl)-4-methylpyridine, quino dichloromethane, chloroform, carbon tetrachloride and 1,2- line, N,N-dimethylaniline, N,N-diethylaniline, DBN, dichloroethane; ethers, such as diethyl ether, diisopropyl DABCO and DBU. Of these, potassium hydroxide is pre ether, tetrahydrofuran and dioxane; aromatic hydrocarbons, ferred. Such as benzene, toluene and nitrobenzene, amides, such as The reaction can take place over a wide range of tempera formamide, N,N-dimethylformamide, N,N-dimethylaceta tures, and the precise reaction temperature is not critical to the mide and hexamethylphosphoric triamide: nitriles, such as invention. The preferred reaction temperature will depend acetonitrile and benzonitrile; sulfoxides, such as dimethyl upon Such factors as the nature of the solvent, and the starting Sulfoxide and Sulfolane, ketones, such as acetone and dieth 25 materials. However, in general, it is convenient to carry out ylketone; water; or mixed solvents thereof. Of these, dimethyl the reactionata temperature of from about 0°C. to about 100° sulfoxide or water is preferred. C. The time required for the reaction may also vary widely, The reaction is carried out in the presence of a halogenating depending on many factors, notably the reaction temperature agent. There is likewise no particular restriction on the nature and the nature of the starting materials and solvent employed. of the halogenating agents used, and any halogenating agent 30 However, provided that the reaction is effected under the commonly used in reactions of this type may equally be used preferred conditions outlined above, a period of from about here. Examples of Such halogenating agents include:Succin 60 minutes to about 48 hours, will usually suffice. imides, such as NBS, NCS; bromine. Of these, NBS is pre The compounds of formula (I), and the intermediates in the ferred. above-mentioned preparation methods can be isolated and 35 purified by conventional procedures, such as distillation, The reaction can take place over a wide range of tempera recrystallization or chromatographic purification. tures, and the precise reaction temperature is not critical to the invention. The preferred reaction temperature will depend Compounds of the invention intended for pharmaceutical upon Such factors as the nature of the solvent, and the starting use may be administered as crystalline or amorphous prod materials. However, in general, it is convenient to carry out ucts. They may be obtained, for example, as solid plugs, the reactionata temperature of from about 0°C. to about 100° 40 powders, or films by methods such as precipitation, crystal C. The time required for the reaction may also vary widely, lization, freeze-drying, spray drying, or evaporative drying. depending on many factors, notably the reaction temperature Microwave or radio frequency drying may be used for this and the nature of the starting materials and solvent employed. purpose. However, provided that the reaction is effected under the Conventional techniques for the preparation/isolation of 45 individual enantiomers include chiral synthesis from a Suit preferred conditions outlined above, a period of from about able optically pure precursor or resolution of the racemate (or 60 minutes to about 24 hours, will usually suffice. the racemate of a salt or derivative) using, for example, chiral (Step G4) high-pressure liquid chromatography (HPLC). In this step, the compound of formula (IX) is prepared by Alternatively, a method of optical resolution of a racemate epoxydation of the compound of formula (XXVIII) 50 (or a racemic precursor) can be appropriately selected from The reaction is normally and preferably effected in the conventional procedures, for example, preferential crystalli presence of solvent. There is no particular restriction on the Zation, or resolution of diastereomeric salts between a basic nature of the solvent to be employed, provided that it has no moiety of the compound of formula (I) and a suitable opti adverse effect on the reaction orthereagents involved and that cally active acid such as tartaric acid. it can dissolve reagents, at least to some extent. Examples of 55 Compounds of the invention intended for pharmaceutical Suitable solvents include: halogenated hydrocarbons, such as use may be administered as crystalline or amorphous prod dichloromethane, chloroform, carbon tetrachloride and 1,2- ucts. They may be obtained, for example, as solid plugs, dichloroethane; ethers, such as diethyl ether, diisopropyl powders, or films by methods such as precipitation, crystal ether, tetrahydrofuran and dioxane; aromatic hydrocarbons, lization, freeze-drying, spray drying, or evaporative drying. Such as benzene, toluene and nitrobenzene, amides, such as 60 Microwave or radio frequency drying may be used for this formamide, N,N-dimethylformamide, N,N-dimethylaceta purpose. mide and hexamethylphosphoric triamide; amines, such as They may be administered alone or in combination with N-methylmorpholine, triethylamine, tripropylamine, tributy one or more other compounds of the invention or in combi lamine, diisopropylethylamine, dicyclohexylamine, N-meth nation with one or more other drugs (or as any combination ylpiperidine, pyridine, 4-pyrrolidinopyridine, N,N-dimethy 65 thereof). Generally, they will be administered as a pharma laniline and N,N-diethylaniline; alcohols, such as methanol, ceutical composition or formulation in association with one ethanol, propanol, 2-propanol and butanol; nitriles, such as or more pharmaceutically acceptable carriers or excipients. US 7,718,809 B2 35 36 The term “carrier' or “excipient' is used herein to describe Tablets also generally contain lubricants such as magne any ingredient other than the compound(s) of the invention. sium Stearate, calcium Stearate, Zinc Stearate, sodium Stearyl The choice of carrier or excipient will to a large extent depend fumarate, and mixtures of magnesium Stearate with sodium on factors such as the particular mode of administration, the lauryl Sulphate. Lubricants generally comprise from about effect of the excipient on solubility and stability, and the 5 0.25 wt % to about 10 wt %, preferably from about 0.5 wt % nature of the dosage form. to about 3 wt % of the tablet. Pharmaceutical compositions suitable for the delivery of Other possible ingredients include anti-oxidants, colou compounds of the present invention and methods for their rants, flavouring agents, preservatives and taste-masking preparation will be readily apparent to those skilled in the art. agents. Such compositions and methods for their preparation may be 10 Exemplary tablets contain up to about 80% drug, from found, for example, in Remington's Pharmaceutical Sci about 10 wt % to about 90 wt % binder, from about 0 wt % to ences, 19th Edition (Mack Publishing Company, 1995). about 85 wt % diluent, from about 2 wt % to about 10 wt % disintegrant, and from about 0.25 wt % to about 10 wt % Oral Administration lubricant. The compounds of the invention may be administered 15 Tablet blends may be compressed directly or by roller to orally. Oral administration may involve Swallowing, so that form tablets. Tablet blends or portions of blends may alterna the compound enters the gastrointestinal tract, or buccal or tively be wet-, dry-, or melt-granulated, melt congealed, or Sublingual administration may be employed by which the extruded before tabletting. The final formulation may com compound enters the blood stream directly from the mouth. prise one or more layers and may be coated or uncoated; it Formulations suitable for oral administration include solid may even be encapsulated. formulations such as, for example, tablets, capsules contain The formulation of tablets is discussed in "Pharmaceutical ing particulates, liquids, or powders, lozenges (including liq Dosage Forms. Tablets, Vol. 1, by H. Lieberman and L. uid-filled), chews, multi- and nano-particulates, gels, Solid Lachman, Marcel Dekker, N.Y., N.Y., 1980 (ISBN 0-8247 Solution, liposome, films (including muco-adhesive), ovules, 6918-X). sprays and liquid formulations. 25 Solid formulations for oral administration may be formu Liquid formulations include, for example, Suspensions, lated to be immediate and/or modified release. Modified Solutions, syrups and elixirs. Such formulations may be release formulations include delayed-, Sustained-, pulsed employed as fillers in soft or hard capsules and typically controlled-, targeted and programmed release. comprise a carrier, for example, water, ethanol, polyethylene 30 Suitable modified release formulations for the purposes of glycol, propylene glycol, methylcellulose, or a suitable oil, the invention are described in U.S. Pat. No. 6,106,864. Details and one or more emulsifying agents and/or Suspending of other suitable release technologies such as high energy agents. Liquid formulations may also be prepared by the dispersions and osmotic and coated particles are to be found reconstitution of a solid, for example, from a sachet. in Verma et al. Pharmaceutical Technology On-line, 25(2), The compounds of the invention may also be used in fast 35 1-14 (2001). The use of chewing gum to achieve controlled dissolving, fast-disintegrating dosage forms such as those release is described in WOOO/35298. described in Expert Opinion in Therapeutic Patents, 11 (6), 981-986 by Liang and Chen (2001). Parenteral Administration For tablet dosage forms, depending on dose, the drug may The compounds of the invention may also be administered make up from about 1 wt % to about 80 wt % of the dosage directly into the blood stream, into muscle, or into an internal form, more typically from about 5 wt % to about 60 wt % of 40 organ. Suitable means for parenteral administration include the dosage form. In addition to the drug, tablets generally intravenous, intraarterial, intraperitoneal, intrathecal, intra contain a disintegrant. Examples of disintegrants include Ventricular, intraurethral, intrasternal, intracranial, intramus Sodium starch glycolate, sodium carboxymethyl cellulose, cular and subcutaneous. Suitable devices for parenteral calcium carboxymethyl cellulose, croScarmellose Sodium, administration include needle (including microneedle) injec crospovidone, polyvinylpyrrolidone, methyl cellulose, 45 tors, needle-free injectors and infusion techniques. microcrystalline cellulose, lower alkyl-substituted hydrox Parenteral formulations are typically aqueous solutions ypropyl cellulose, starch, pregelatinized starch and sodium which may contain excipients such as salts, carbohydrates alginate. Generally, the disintegrant will comprise from about and buffering agents (preferably to a pH of from about 3 to 1 wt % to about 25 wt %, preferably from about 5 wt % to about 9), but, for some applications, they may be more Suit about 20 wt % of the dosage form. 50 ably formulated as a sterile non-aqueous Solution or as a dried Binders are generally used to impart cohesive qualities to a form to be used in conjunction with a suitable vehicle such as tablet formulation. Suitable binders include microcrystalline sterile, pyrogen-free water. cellulose, gelatin, Sugars, polyethylene glycol, natural and The preparation of parenteral formulations under sterile synthetic gums, polyvinylpyrrolidone, pregelatinized starch, 55 conditions, for example, by lyophilisation, may readily be hydroxypropyl cellulose and hydroxypropyl methylcellu accomplished using standard pharmaceutical techniques well lose. Tablets may also contain diluents, such as lactose known to those skilled in the art. (monohydrate, spray-dried monohydrate, anhydrous and the The solubility of compounds of formula (I) used in the like), mannitol, Xylitol, dextrose, Sucrose, Sorbitol, microc preparation of parenteral Solutions may be increased by the rystalline cellulose, starch and dibasic calcium phosphate 60 use of appropriate formulation techniques, such as the incor dihydrate. poration of Solubility-enhancing agents. Tablets may also optionally comprise Surface-active Formulations for parenteral administration may be formu agents, such as Sodium lauryl Sulfate and polysorbate 80, and lated to be immediate and/or modified release. Modified glidants such as silicon dioxide and talc. When present, Sur release formulations include delayed-, Sustained-, pulsed face active agents may comprise from about 0.2 wt % to about 65 controlled-, targeted and programmed release. Thus com 5 wt % of the tablet, and glidants may comprise from about pounds of the invention may be formulated as a solid, semi 0.2 wt % to about 1 wt % of the tablet. Solid, or thixotropic liquid for administration as an implanted US 7,718,809 B2 37 38 depot providing modified release of the active compound. water, ethanol and sodium chloride. Alternative solvents Examples of Such formulations include drug-coated Stents which may be used instead of propylene glycol include glyc and PGLA microspheres. erol and polyethylene glycol. Topical Administration Suitable flavours, such as menthol and levomenthol, or The compounds of the invention may also be administered Sweeteners, such as Saccharin or saccharin Sodium, may be topically to the skin or mucosa, that is, dermally or transder added to those formulations of the invention intended for mally. Typical formulations for this purpose include gels, inhaled/intranasal administration. Formulations for inhaled/ hydrogels, lotions, solutions, creams, ointments, dusting intranasal administration may be formulated to be immediate powders, dressings, foams, films, skin patches, wafers, and/or modified release using, for example, poly(DL-lactic implants, sponges, fibres, bandages and microemulsions. 10 coglycolic acid) (PGLA). Modified release formulations Liposomes may also be used. Typical carriers include alco include delayed-, Sustained-, pulsed-, controlled-, targeted hol, water, mineral oil, liquid petrolatum, white petrolatum, and programmed release. glycerin, polyethylene glycol and propylene glycol. Penetra In the case of dry powder inhalers and aerosols, the dosage tion enhancers may be incorporated—see, for example, J unit is determined by means of a valve which delivers a Pharm Sci, 88 (10),955-958 by Finnin and Morgan (October 15 metered amount. Units in accordance with the invention are 1999). typically arranged to administer a metered dose or “puff Other means of topical administration include delivery by containing from about 1 to about 100 ug of the compound of electroporation, iontophoresis, phonophoresis, Sonophoresis formula (I). The overall daily dose will typically be in the and microneedle or needle-free (e.g. PowderjectTM, range about 50 ug to about 20 mg which may be administered BiojectTM, etc.) injection. in a single dose or, more usually, as divided doses throughout Formulations for topical administration may beformulated the day. to be immediate and/or modified release. Modified release Rectal/Intravaginal Administration formulations include delayed-, Sustained-, pulsed-, con The compounds of the invention may be administered rec trolled-, targeted and programmed release. tally or vaginally, for example, in the form of a Suppository, 25 pessary, or enema. Cocoa butter is a traditional Suppository Inhaled/Intranasal Administration base, but various alternatives may be used as appropriate. The compounds of the invention can also be administered Formulations for rectal/vaginal administration may be for intranasally or by inhalation, typically in the form of a dry mulated to be immediate and/or modified release. Modified powder (either alone, as a mixture, for example, in a dry blend release formulations include delayed-, Sustained-, pulsed with lactose, or as a mixed component particle, for example, 30 mixed with phospholipids, such as phosphatidylcholine) controlled-, targeted and programmed release. from a dry powder inhaler or as an aerosol spray from a Ocular/Aural Administration pressurized container, pump, spray, atomiser (preferably an The compounds of the invention may also be administered atomiser using electrohydrodynamics to produce a fine mist), directly to the eye or ear, typically in the form of drops of a or nebuliser, with or without the use of a suitable propellant, 35 micronised Suspension or solution in isotonic, pH-adjusted, such as 1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluo sterile saline. Otherformulations suitable for ocular and aural ropropane. For intranasal use, the powder may comprise a administration include ointments, biodegradable (e.g. bioadhesive agent, for example, chitosan or cyclodextrin. absorbable gel sponges, collagen) and non-biodegradable The pressurized container, pump, spray, atomizer, or nebu (e.g. silicone) implants, wafers, lenses and particulate or liser contains a solution or Suspension of the compound(s) of 40 vesicular systems. Such as niosomes or liposomes. A polymer the invention comprising, for example, ethanol, aqueous etha Such as crossed-linked polyacrylic acid, polyvinylalcohol, nol, or a suitable alternative agent for dispersing, solubilising, hyaluronic acid, a cellulosic polymer, for example, hydrox or extending release of the active, a propellant(s) as solvent ypropylmethylcellulose, hydroxyethylcellulose, or methyl and an optional Surfactant, such as Sorbitan trioleate, oleic cellulose, or a heteropolysaccharide polymer, for example, acid, or an oligolactic acid. 45 gelangum, may be incorporated together with a preservative, Prior to use in a dry powder or Suspension formulation, the Such as benzalkonium chloride. Such formulations may also drug product is micronised to a size suitable for delivery by be delivered by iontophoresis. inhalation (typically less than 5 microns). This may be Formulations for ocular/aural administration may be for achieved by any appropriate comminuting method, such as mulated to be immediate and/or modified release. Modified spiral jet milling, fluid bed jet milling, Supercritical fluid 50 release formulations include delayed-, Sustained-, pulsed processing to form nanoparticles, high pressure homogeniza controlled-, targeted, or programmed release. tion, or spray drying. Other Technologies Capsules (made, for example, from gelatin or HPMC), The compounds of the invention may be combined with blisters and cartridges for use in an inhaler or insufflator may soluble macromolecular entities, such as cyclodextrin and be formulated to contain a powder mix of the compound of 55 suitable derivatives thereof or polyethylene glycol-contain the invention, a Suitable powder base Such as lactose or starch ing polymers, in order to improve their solubility, dissolution and a performance modifier Such as 1-leucine, mannitol, or rate, taste-masking, bioavailability and/or stability for use in magnesium Stearate. The lactose may be anhydrous or in the any of the aforementioned modes of administration. form of the monohydrate, preferably the latter. Other suitable Drug-cyclodextrin complexes, for example, are found to be excipients include dextran, glucose, maltose, Sorbitol, Xylitol, 60 generally useful for most dosage forms and administration fructose. Sucrose and trehalose. routes. Both inclusion and non-inclusion complexes may be A Suitable solution formulation for use in anatomiser using used. As an alternative to direct complexation with the drug, electrohydrodynamics to produce a fine mist may contain the cyclodextrin may be used as an auxiliary additive, i.e. as from about 1 ug to about 20 mg of the compound of the a carrier, diluent, or solubiliser. Most commonly used for invention per actuation and the actuation Volume may vary 65 these purposes are alpha-, beta- and gamma-cyclodextrins, from about 1 ul to about 100 ul. Atypical formulation may examples of which may be found in. WO 91/11 172, WO comprise a compound of formula (I), propylene glycol, Sterile 94/O2518 and WO 98/55148. US 7,718,809 B2 39 40 Kit-Of-Parts (ix) GABA agonists, e.g. baclofen and AZD-3355; Inasmuch as it may be desirable to administer a combina (x) GABA antagonists, e.g. GAS-360 and SGS-742: tion of active compounds, for example, for the purpose of (xi) calcium channel blockers, e.g. aranidipine, lacidipine, treating aparticular disease or condition, it is within the scope falodipine, azelnidipine, clinidipine, lomerizine, dilt of the present invention that two or more pharmaceutical 5 iazem, gallopamil, efonidipine, nisoldipine, amlodipine, compositions, at least one of which contains a compound in lercanidipine, bevantolol, nicardipine, isradipine, benid accordance with the invention, may conveniently be com ipine, Verapamil, nitrendipine, barnidipine, propafenone, bined in the form of a kit suitable for coadministration of the manidipine, bepridil, nifedipine, nilvadipine, nimodipine compositions. and fasudil; Thus the kit of the invention comprises two or more sepa 10 (xii) dopamine antagonists, e.g. metoclopramide, domperi rate pharmaceutical compositions, at least one of which con done and levosulpiride: tains a compound of formula (I) in accordance with the inven (xiii) Tachykinin (NK) antagonists, particularly NK-3, NK-2 tion, and means for separately retaining said compositions, and NK-1 antagonists, e.g.: nepadutant, Saredutant, talnet such as a container, divided bottle, or divided foil packet. An ant, (O.R.9R)-7-3,5-bis(trifluoromethyl)benzyl-8.9.10, example of such a kit is the familiar blister pack used for the 15 11-tetrahydro-9-methyl-5-(4-methylphenyl)-7H-1,4-dia packaging of tablets, capsules and the like. Zocino.2.1-g1.7naphthridine-6-13-dione (TAK-637), The kit of the invention is particularly suitable for admin 5-(2R.3S)-2-[(1R)-1-3,5-bis(trifluoromethyl)phenyl istering different dosage forms, for example, oral and ethoxy-3-(4-fluoropheny)-4-morpholinyl)methyl-1,2-di parenteral, for administering the separate compositions at hydro-3H-1,2,4-triazol-3-one (MK-869), lanepitant, dapi different dosage intervals, or for titrating the separate com tant and 3-2-methoxy-5-(trifluoromethoxy)phenyl positions against one another. To assist compliance, the kit methylamino-2-phenyl-piperidine (2S,3S); typically comprises directions for administration and may be (xiv) Helicobacter pylori infection agents, e.g., clarithromi provided with a so-called memory aid. cyn, roXithromycin, rokitamycin, flurithromycin, tellithro Dosage mycin, amoxicillin, amplicillin, temocillin, bacampicillin, For administration to human patients, the total daily dose 25 aspoxicillin, Sultamicillin, piperacillin, lenampicillin, tet of the compounds of the invention is typically in the range of racycline, metronidazole, bithmuth citrate and bithmuth about 0.05 mg to about 100 mg depending, of course, on the Subsalicylate. mode of administration, preferred in the range of about 0.1 (XV) nitric oxide synthase inhibitors, e.g. GW-274150, tillargi mg to about 50 mg and more preferred in the range of about nine, P54, guanidioethyldisulfide and nitroflurbiprofen; 0.5 mg to about 20 mg. For example, oral administration may 30 (xvi) vanilloid receptor 1 antagonists, e.g. AMG-517 and require a total daily dose of from about 1 mg to about 20 mg. GW-705498: while an intravenous dose may only require from about 0.5 (Xvii) muscarinic receptor antagonists, e.g. trospium, Solif mg to about 10 mg. The total daily dose may be administered enacin, tolterodine, tiotropium, cimetropium, OXitropium, in single or divided doses. ipratropium, tiquizium, dalifenacin and imidafenacin; These dosages are based on an average human Subject 35 (Xviii) calmodulinantagonists, e.g. squalamine and DY-9760; having a weight of about 65 kg to about 70 kg. The physician (xix) potassium channel agonists, e.g. pinacidil, tilisolol, nic will readily be able to determine doses for subjects whose orandil, NS-8 and retigabine; weightfalls outside this range. Such as infants and the elderly. (XX) beta-1 agonists, e.g. dobutamine, denopamine, Xamot An acid pump antagonist of the present invention may be erol, denopamine, docarpamine and Xamoterol; usefully combined with another pharmacologically active 40 (XXi) beta-2 agonists, e.g. salbutamol, terbutaline, arformot compound, or with two or more other pharmacologically erol, meluadrine, mabuterol, ritodrine, fenoterol, clen active compounds, particularly in the treatment of gastroe buterol, formoterol, procaterol, tulobuterol, pirbuterol, sophageal reflux disease. For example, an acid pump antago bambuterol, tulobuterol, dopexamine and levosalbutamol; nist, particularly a compound of the formula (I), or a pharma 45 (XXii) beta agonists, e.g. isoproterenol and terbutaline; ceutically acceptable salt thereof, as defined above, may be (XXiii) alpha 2 agonists, e.g. clonidine, medetomidine, lofexi administered simultaneously, sequentially or separately in dine, moxonidine, tizanidine, guanfacine, guanabenz, tal combination with one or more agents selected from: ipexole and dexmedetomidine, (i) histamine H receptor antagonists, e.g. ranitidine, lafuti (XXiv) endthelin A antagonists, e.g. bonsetan, atrasentan, dine, nizatidine, cimetidine, famotidine and roXatidine; 50 ambrisentan, claZosentan, sitaxsentan, fandoSentan and (ii) proton pump inhibitors, e.g. omeprazole, esomeprazole, darusentan; pantoprazole, rabeprazole, tenatoprazole, ilaprazole and (XXV) opioid Lagonists, e.g. morphine, fentanyl and lopera lansoprazole; mide; (iii) oral antacid mixtures, e.g. Maalox R, AludroxR) and (XXvi) opioid Lantagonists, e.g. naloxone, buprenorphine and Gaviscon R; 55 alvimopan: (iv) mucosal protective agents, e.g. polaprezinc, ecabet (XXvii) motilinagonists, e.g. erythromycin, mitemcinal, SLV Sodium, rebamipide, teprenone, cetraxate, Sucralfate, chlo 305 and atilmotin; ropylline-copper and plaunotol; (XXviii) ghrelin agonists, e.g. capromorelin and TZP-101; (v) anti-gastric agents, e.g. Anti-gastrin vaccine, itriglumide (xxix) AchE release stimulants, e.g. Z-338 and KW-5092; and Z-360; 60 (XXX) CCK-B antagonists, e.g. itriglumide, YF-476 and (vi) 5-HT antagonists, e.g. dolasetron, palonosetron, alos S-0509; etron, aZasetron, ramosetron, mitrazapine, granisetron, tro (xxxi) glucagon antagonists, e.g. NN-2501 and A-77.0077. pisetron, E-3620, ondansetron and indisetron; (vii) 5-HT agonists, e.g. tegaserod, mosapride, cinitapride Method for Assessing Biological Activities: and OXtriptane; 65 The acid pump inhibitory activity and other biological (viii) laxatives, e.g. TrifybaR), Fybogel(R), Konsyl(R), Isogel(R), activities of the compounds of this invention were determined Regulan R., Celevac(R) and Normacol R; by the following procedures US 7,718,809 B2 41 42 Preparation of Gastric Vesicles from Fresh Porcine Stomachs The porcine gastric vesicles for Porcine gastric H/K"- TABLE 1-continued ATPase inhibition assays were prepared from mucous mem brane in fresh porcine stomachs by homogenization with a Example No. ICso (IM) tight-fitted polytetrafluoroethylene (Teflone(R) homogenizer 11 O43 in 0.25 M sucrose at 4°C. The crude pellet was removed with 12 O.67 13 O.12 centrifugation at 20,000 g for 30 min. Then supernatant was 14 O.98 centrifuged at 100,000 g for 30 min. The resulting pellet was 15 O.95 re-suspended in 0.25 M sucrose, and then subjected to density 16 O.14 gradient centrifugation at 132,000 g for 90 min. The gastric 10 17 O.34 vesicles were collected from interface on 0.25 M sucrose 18 O.24 19 O.68 layer containing 7% FicollTM PM400(Amersham Bio 2O O.61 Sciences). This procedure was performed in a cold room. 21 O49 22 O.18 Ion-Leaky Porcine Gastric H+/K+-ATPase Inhibition 15 23 0.44 Ion-leaky porcine gastric H/K-ATPase inhibition was 24 O.62 measured according to the modified method described in 25 O.S9 Biochemical Pharmacology, 1988, 37, 2231-2236. 26 O48 The isolated vesicles were lyophilized, and then kept in All the tested compounds showed acid pump antagonistic activity. deep-freezeruntiluse. Forenzyme assay, lyophilized vesicles were reconstituted with 3 mM MgSO containing 40 mM Canine Kidney Na/K"-ATPase Inhibition Bis-tris (pH 6.4 at 37° C.). The powdered canine kidney Na/K-ATPase (Sigma) was Enzyme reaction was performed incubating 5 mM KC1, 3 reconstituted with 3 mM MgSO containing 40 mM Tris (pH mM NaATP, 3 mM MgSO and 1.0 g of reconstituted 7.4 at 37° C.). Enzyme reaction was performed incubating vesicles for 30 minutes at 37°C. in a final 60 ul of reaction 25 100 mMNaCl, 2 mMKC1, 3 mMNaATP.3 mMMgSO and mixture (40 mM Bis-tris, pH 6.4) with or without the test 12 lug of enzyme for 30 minutes at 37° C. in a final 60 ul of compound. Enzyme reaction was stopped by adding 10% reaction mixture (40 mM Tris, pH 7.4) with or without the test sodium dodecyl sulphate (SDS). Released inorganic phos compound. Enzyme reaction was stopped by adding 10% phate from ATP was detected by incubation with mixture of 1 SDS. Released inorganic phosphate from ATP was detected part of 35 mMammonium molybdate tetrahydrate in 15 mM 30 by incubating with mixture of 1 part of 35 mM ammonium Zinc acetate hydrate and 4 parts of 10% ascorbic acid (pH molybdate tetrahydrate in 15 mM Zinc acetate hydrate and 4 5.0), resulting in phosphomolybdate, which has optical den parts of 10% ascorbic acid (pH 5.0), resulting in phosphomo sity at 750 nm. All example compounds showed potent inhibi lybdate, which has optical density at 750 nm. tory activity. Inhibition of Acid Secretion in the Gastric Lumen-Perfused Ion-tight Porcine Gastric H+/K+-ATPase Inhibition 35 Rat Ion-tight porcine gastric H/K"-ATPase inhibition was Acid secretion in the gastric lumen-perfused rat was mea measured according to the modified method described in sured according to Watanabe et al. Watanabe K et al., J. Biochemical Pharmacology, 1988, 37, 2231-2236. Physiol. (Paris) 2000; 94: 111-116). The isolated vesicles were kept in deep-freezer until use. Male Sprague-Dawley rats, 8 weeks old, deprived of food For enzyme assay, vesicles were diluted with 3 mM MgSO 40 for 18 hours before the experiment with free access to water, containing 5 mM Tris (pH 7.4 at 37° C). were anesthetized with urethane (1.4 g/kg, i.p.) and trache Enzyme reaction was performed incubating 150 mM KCl, otomized. After a middle abdominal incision, a dual polyeth 3 mMNaATP.3 mMMgSO 15uM valinomycin and 3.0 ug ylene cannula was inserted into the forestomachand the stom of vesicles for 30 minutes at 37°C. in a final 60 ul of reaction ach was perfused with saline (37° C. pH 5.0) at a rate of 1 mixture (5 mM Tris, pH 7.4) with or without the test com 45 ml/min. The acid output in the perfusate was determined at 5 pound. Enzyme reaction was stopped by adding 10% SDS. minutes interval by titration with 0.02 N. NaOH to pH 5.0. Released inorganic phosphate from ATP was detected by After the determination of basal acid secretion for 30 min, the incubating with mixture of 1 part of 35 mM ammonium acid secretion was stimulated by a continuous intravenous molybdate tetrahydrate in 15 mM Zinc acetate hydrate and 4 infusion of pentagastrin (16 g/kg/h). The test compounds parts of 10% ascorbic acid (pH 5.0), resulting in phosphomo 50 were administered by an intravenous bolus injection or lybdate, which has optical density at 750 nm. intraduodenal administration after the stimulated acid secre The results of ICs values of the inhibitory activity for the tion reached a plateau phase. The acid secretion was moni compounds of following examples are shown in Table 1. tored after the administration. 55 The activity was evaluated either inhibition of total acid TABLE 1. secretion from 0 hours to 1.5 or 3.5 hours after administration or the maximum inhibition after administration. Example No. ICso (LIM) Inhibition of Gastric Acid Secretion in the Heidenhain Pouch O.S6 Dog O.069 0.44 60 Male Beagle dogs weighing 7-15 kg with Heidenhain 0.44 pouch Heidenhain R: Arch Ges Physiol. 1879; 19: 148-167 O.O90 were used. The animals were allowed to recover from surgery O.21 O.22 for at least three weeks before the experiments. The animals O.18 were kept at a 12 hour light-dark rhythm, housed singly. They O.96 65 received standard food once daily at 11:00 a.m. and tap water 1 O.32 adlibitum, and were fasted overnight prior to the experiment, with free access to water. Gastric juice samples were col US 7,718,809 B2 43 44 lected throughout the experiment by gravity drainage every ml apical buffer and 1.0 ml basolateral buffer for 0.5 hour at 15 min. Acidity in the gastric juice was measured by titration 37° C. in a shaker water bath at 50 cycles/min. The apical to the end point of pH 7.0. Acid secretion was stimulated by buffer consisted of Hanks Balanced Salt Solution, 25 mM a continuous intravenous infusion of histamine (80 ug/kg/h). D-glucose monohydrate, 20 mM 2-morpholinoethanesul Oral or intravenous bolus administration of the test com phonic acid (MES) Biological Buffer, 1.25 mM CaCl and 0.5 pounds was done 90 minutes after commencement of the mM MgCl (pH 6.5). The basolateral buffer consisted of histamine infusion. The acid secretion was monitored after Hanks Balanced Salt Solution, 25 mM D-glucose monohy the administration. The activity was evaluated by the maxi drate, 20 mM 2-4-(2-hydroxyethyl)-1-piperazinylethane mum inhibition relative to the corresponding control value. sulfonic acid (HEPES) Biological Buffer, 1.25 mM CaCl, The compound of Example 6 showed a good inhibitory 10 and 0.5 mM MgCl (pH 7.4). At the end of the preincubation, activity. the media was removed and test compound solution (10 uM) Human Dofetilide Binding in buffer was added to the apical compartment. The inserts Human ether a-go-go related gene (HERG) transfected were moved to wells containing fresh basolateral buffer at 1 HEK293S cells were prepared and grown in-house. Cell paste hour. Drug concentration in the buffer was measured by of HEK-293 cells expressing the HERG product can be sus 15 LC/MS analysis. pended in 10-fold volume of 50 mM Tris buffer adjusted at pH Flux rate (F, mass/time) was calculated from the slope of 7.5 at 25°C. with 2M HCl containing 1 mM MgCl, 10 mM cumulative appearance of Substrate on the receiver side and apparent permeability coefficient (P)cipp was calculated from KC1. The cells were homogenized using a Polytron homog the following equation. enizer (at the maximum power for 20 seconds) and centri fuged at 48,000 g for 20 minutes at 4°C. The pellet was resuspended, homogenized and centrifuged once more in the same manner. The resultant Supernatant was discarded and where SA is surface area for transport (0.3 cm), VD is the the final pellet was resuspended (10-fold volume of 50 mM donor volume (0.3 ml), MD is the total amount of drug on the Tris buffer) and homogenized at the maximum power for 20 donor side at t=0. All data represent the mean of 2 inserts. seconds. The membrane homogenate was aliquoted and 25 Monolayer integrity was determined by Luciferyellow trans stored at -80° C. until use. An aliquot was used for protein port. concentration determination using a Protein Assay Rapid Kit Half-life in Human Liver Microsomes (HLM) (wako) and Spectra max plate reader (Wallac). All the Test compounds (1 uM) were incubated with 3.3 mM manipulation, stock solution and equipment were kept on ice 30 MgCl, and 0.78 mg/mL HLM (HL 101) in 100 mM potassium at all times. For Saturation assays, experiments were con phosphate buffer (pH 7.4) at 37°C. on the 96-deep well plate. ducted in a total volume of 200ul. Saturation was determined The reaction mixture was split into two groups, a non-P450 by incubating 36 ul of H-dofetilide, and 160 ul of mem and a P450 group. NADPH was only added to the reaction brane homogenates (20-30 ug protein per well) for 60 min mixture of the P450 group. An aliquot of samples of P450 utes at room temperature in the absence or presence of 10M 35 group was collected at 0, 10, 30, and 60 minutes time point, dofetilide at final concentrations (4 ul) for total or nonspecific where 0 minutes time point indicated the time when NADPH binding, respectively. All incubations were terminated by was added into the reaction mixture of P450 group. An aliquot rapid vacuum filtration over PEI soaked glass fiber filter of samples of non-P450 group was collected at -10 and 65 papers using Skatron cell harvester followed by two washes minutes time point. Collected aliquots were extracted with with 50 mM Tris buffer (pH 7.4 at 25° C). Receptor-bound 40 acetonitrile solution containing an internal standard. The pre radioactivity was quantified by liquid Scintillation counting cipitated protein was spun down in centrifuge (2000 rpm, 15 using Packard LS counter. min). The compound concentration in Supernatant was mea For the competition assay, compounds were diluted in 96 sured by LC/MS/MS system. well polypropylene plates as 4-point dilutions in semi-log The half-life value was obtained by plotting the natural format. All dilutions were performed in DMSO first and then 45 logarithm of the peak area ratio of compounds/internal stan transferred into 50 mM Tris buffer (pH 7.4 at 25°C.) contain dard versus time. The slope of the line of best fit through the ing 1 mM MgCl, 10 mM KCl so that the final DMSO con points yields the rate of metabolism (k). This was converted to centration became equal to 1%. Compounds were dispensed a half-life value using following equations: in triplicate in assay plates (4 Jul). Total binding and nonspe cific binding wells were set up in 6 wells as vehicle and 10LM 50 dofetilide at final concentration, respectively. The radioligand In vitro Drug-Drug Interaction Studies for Five Major CYPs was prepared at 5.6x final concentration and this solution was (fDDI) added to each well (36 ul). The assay was initiated by addition CYP1A2 Test compounds (3 uM) were pre-incubated with of YSipoly-L-lysine SPA beads (50 ul, 1 mg/well) and mem recombinant CYP1A2 (Baculosome lot #21198 Invitro branes (110 ul, 20 g/well). Incubation was continued for 60 55 gen, 50 pmol P450/ml) in 100 mM K"Phosphate Buffer minutes at room temperature. Plates were incubated for a (pH 7.4) and 10 uMVivid blue 1A2 probe (Invitrogen) as a further 3 hours at room temperature for beads to settle. Recep substrate for 5 minutes at 30° C. Reaction was initiated by tor-bound radioactivity was quantified by counting Wallac adding a solution of a warmed NADPH-regenerating sys MicroBeta plate counter. tem A, which consists of 0.50 mM NADP and 10 mM Caco-2 Permeability 60 MgCl, 6.2 mM DL-Isocitric acid and 0.5 U/ml Isocitric Caco-2 permeability was measured according to the Dehydrogenase (ICD). Plates were placed in the plate method described in Shiyin Yee, Pharmaceutical Research, reader at 30°C. and were taken readings every 1.5 minutes, 763 (1997). with a 10 second shake in between each reading for 15 Caco-2 cells were grown on filter supports (Falcon HTS cycles. Wavelengths of excitation/emission were 408/465 multiwell insert system) for 14 days. Culture medium was 65 nm, respectively. removed from both the apical and basolateral compartments CYP2C9 Test compounds (3 uM) were pre-incubated with and the monolayers were preincubated with pre-warmed 0.3 recombinant CYP2C9 (Baculosome lot #20967 Invitro US 7,718,809 B2 45 46 gen, 50 pmol P450/ml) in 100 mM K"Phosphate Buffer and patch pipettes which have a resistance of 1-3 MOhm (pH 7.4) and 30 uMMFC probe (Gentest) as a substrate for when filled with the standard internal solution of the follow 5 minutes at 37° C. Reaction was initiated by adding a ing composition; (mM); KC1, 130; MgATP, 5: MgCl, 1: solution of the warmed NADPH-regenerating system A. HEPES, 10: EGTA5, pH 7.2 with KOH. Only those cells with Plates were placed in the plate reader at 37° C. and were access resistances below 10 MOhm and seal resistances over taken readings every 2.0 minutes, with a 10 second shake in 1 GOhm are accepted for further experimentation. Series between each reading for 15 cycles. Wavelengths of exci resistance compensation is applied up to a maximum of 80% tation/emission were 408/535 nm, respectively. without any leak subtraction. Following the achievement of CYP2C19 Test compounds (3 uM) were pre-incubated with whole cell configuration and sufficient time for cell dialysis recombinant CYP2C19 (Baculosome lot #20795 Invitro 10 with pipette solution (>5 min), the membrane is depolarized gen, 5 pmol P450/ml) in 100 mMK"Phosphate Buffer (pH from a holding potential of -80 mV to +30 mV for 1000 ms 7.4) and 10 uMVivid blue 2C19 probe (Invitrogen) as a followed by a descending voltage ramp (rate 0.5 mV msec') substrate for 5 minutes at 37°C. Reaction was initiated by back to the holding potential. This depolarization and ramp is adding a solution of the warmed NADPH-regenerating applied to the cells continuously every 4 seconds (0.25 Hz). system A. Plates were placed in the plate reader at 37° C. 15 and were taken readings every 1.5 minutes with a 10 sec The amplitude of the peak current elicited around -40 mV ond shake in between each reading for 15 cycles. Wave during the ramp is measured. Once stable evoked current lengths of excitation/emission were 408/465 nm, respec responses of minimal changes in the amplitude are obtained tively. in the external Solution, the test compound is applied for CYP2D6 Test compounds (3 uM) were pre-incubated with 10-20 minutes with multiple dosing in a single cell. The cells recombinant CYP2D6 (Baculosome lot #21248 Invitro are also exposed to high dose of dolfetilide (5uM), a specific gen, 20 pmol P450/ml) in 100 mM K"Phosphate Buffer IKr blocker, to evaluate the insensitive endogenous current. (pH 7.4) and 1 uM 3-2-(N,N-diethyl-N-methylammo All experiments are performed at 23+/-1° C. Evoked nium)ethyl-7-methoxy-4-methylcoumarin (AMMC) membrane currents are recorded online on a computer, fil probe (Gentest) as a substrate for 5 minutes at 37° C. 25 tered at 500-1000Hz (Bessel-3 dB) and sampled at 1-2 KHZ. Reaction was initiated by adding a solution of a warmed Osmolarity and pH change induced by the test compound in NADPH-regenerating system B, which consists of 0.03 external Solution will be examined at the highest concentra mM NADP and 10 mM MgCl, 6.2 mMDL-Isocitric acid tion. and 0.5 U/mL ICD. Plates were placed in the plate reader at The arithmetic mean of these ten values of peak current is 37° C. and were taken readings every 2.0 minutes with a 10 30 calculated under control conditions and in the presence of second shake in between each reading for 15 cycles. Wave drug. Percent decrease of Iyin each experiment is obtained by lengths of excitation/emission were 400/465 nm, respec the normalized current value using the following formula: tively. Iy (I-I,)/(I-I)x100, where I is the mean current value CYP3A4 Test compounds (3 uM) were pre-incubated with under control conditions, I is the mean current value in the recombinant CYP3A4 (Baculosome lot #20814 Invitro 35 presence of test compound and I is the mean current value gen, 5 pmol P450/ml) in 100 mMK"Phosphate Buffer (pH in dolfetilide application. Separate experiments are performed 7.4) and 2 uMVivid Red probe (Invitrogen) as a substrate and pooled data of arithmetic mean from each experiment is for 5 minutes at 30° C. Reaction was initiated by adding a defined as the result of the study. solution of the warmed NADPH-regenerating system A. Plates were placed in the plate reader at 30° C. and were 40 hERG Patch Clamp Assay taken readings minimum intervals with a 10 second shake To determine the potential of compounds to inhibit the in between each reading for 15 cycles. Wavelengths of hERG channel, the cloned counterpart of the rapidly inacti excitation/emission were 530/595 nm, respectively. Vating delayed rectifier potassium current (IKr). Drug-drug interaction was evaluated by the rate of metabo HEK293 cells stably expressing the hERG channel were lite formation calculated with a slope (Time vs. Fluorescence 45 used in whole-cell patch clamp electrophysiology studies at units) in the linear region or the percentage of inhibition by ambient temperature (26.5-28.5° C.). The methodology for test compounds calculated by the following equation. stable transfection of this channel in HEK293 cells can be Inhibition%={(1-1)/v}* 100, wherein v is a rate of found elsewhere (Zhou et al 1998, Biophysical Journal, 74, control reaction (no test compounds) and v, is a pp230-241). The solutions used for experimentation were rate of reaction in the presence of test compound. 50 standard extracellular solution of the following composition (mM); NaCl, 137: KC1, 4: CaCl, 1.8; MgCl, 1: Glucose, 10; Irrero, Assay HEPES, 10; pH 7.4+0.05 with NaOH/HC1; and standard Human ether a-go-go related gene (HERG) transfected intracellular solution of the following composition (mM); HEK293 cells are prepared and cultured in-house. The meth KC1, 130; MgCl, 1: HEPES, 10; EGTA, 5; MgATP, 5; pH odology for stable transfection of this channel in HEK cells 55 7.2+0.05 with KOH. The voltage protocol applied was can be found elsewhere (Z.Zhou et al., 1998, Biophysical designed to activate the hERG channel and allow the mea journal, 74, 230-241). On the day of experimentation, the surement of drug block of the channel and is as follows. First cells are harvested from culture flasks and stored as cell the membrane potential was stepped from a holding potential Suspension in a standard external solution (see below of its of -80 mV to +30 mV for 1 s. This was followed by a composition). in the room atmosphere of 23° C. Cells are 60 descending Voltage ramp at a rate of 0.5 mV/ms back to studied between 0.5-5 hours after harvest. holding potential of -80 mV and the peak outward current HERG currents are studied using a standard patch clamp observed during the repolarizing ramp was measured. This technique of the whole-cell mode. During the experiment, the protocol was evoked repeatedly every 4 seconds (0.25 Hz). cells are superfused with a standard external solution of the After establishing a stable baseline period in the presence of following composition:(mM) NaCl, 130; KC1, 4: CaCl2, 2: 65 vehicle (0.1% V/v DMSO), four increasing concentrations of MgCl, 1: Glucose, 10: HEPES, 5; pH 7.4 with NaOH. test compound were then bath-applied sequentially until the Whole-cell recordings is made using a patch clamp amplifier response reached steady-state or 10 minutes (whichever US 7,718,809 B2 47 48 occurred first). 10 micromol/L dofetilide was used at the end taurocholate (NaTC) and 1.06 mg of 1-palmitoyl-2-oleyl-L- of each experiment as an internal positive control and to phosphatidylcholine (POPC) in 1 mL of PBS (pH 6.5). define maximum block. Estimation of Hepatic Clearance Using the Metabolic Stabil Bioavailability in Rat ity in Human Hepatocytes Adult rats of the Sprague-Dawley strain were used. One to Tested compounds (1 uM) were incubated statically with two days prior to the experiments all rats were prepared by hepatocytes from human at 37°C. in a 95% air/5% CO, with cannulation of the right jugular vein under anesthesia. The target cell density of 0.5x10 cells/mlanda total volume of 50 cannula was exteriorized at the nape of the neck. Blood LL. Incubation was stopped at each time point by the addition samples (0.2-0.3 mL) were drawn from the jugular vein at 10 of ice-cold acetonitrile (ACN). Aliquots of samples were intervals up to 24 hours after intravenous or oral administra mixed with 10% ACN containing an internal standard for tions of the test compound. The samples were frozen until LC/MS/MS analysis. After samples were sonicated for 10 analysis. Bioavailability was assessed by calculating the quo minutes, samples were centrifuged at 2,000 rpm for 15 min utes, and then the Supernatant was transferred to the other tient between the area under plasma concentration curve plates for analysis. The compound concentrations in Super (AUC) following oral administration or intravenous admin 15 istration. natant were measured by LC/MS/MS system. The disappearance rates of tested compounds were Bioavailability in Dog obtained by plotting the common logarithm of the peak area Adult Beagle dogs were used. Blood samples (0.2-0.5 mL) ratio of compounds/internal standard versus time. The slope were drawn from the cephalic vein at intervals up to 24 hours of the line of best fit through the points yielded the rate of after intravenous or oral administrations of the test com metabolism (k). This value was scaled to take hepatocellu pound. The samples were frozen until analysis. Bioavailabil larity, liver and body weight into account to give an intrinsic ity was assessed by calculating the quotient between the area clearance value (CL) in ml/min/kg as illustrated in Equa under plasma concentration curve (AUC) following oral tion 1. Hepatic clearance (CL) was predicted from this administration or intravenous administration. intrinsic clearance value using the parallel tube model as 25 shown in Equation 2. The predicted clearance divided by the Plasma Protein Binding hepatic blood flow (Q) afforded the extraction ratio (E) Plasma protein binding of the test compound (1 uM) was (Equation 3). measured by the method of equilibrium dialysis using kx (g liver/kg body weight)x(ml incubation number 96-well plate type equipment. Spectra-Por(R), regenerated cel of cells in incubation)x(cellsig liver) Equation 1: lulose membranes (molecular weight cut-off 12,000-14,000, 30 22 mmx120 mm) were soaked for over night in distilled CL-9x{1-exp(-CL/Q)} Equation 2: water, then for 20 minutes in 30% ethanol, and finally for 15 minutes in dialysis buffer (Dulbecco's phosphate buffered E-CL/9, Equation 3: saline, pH 7.4). Frozen plasma of human, Sprague-Dawley rats, and Beagle dogs were used. The dialysis equipment was 35 Wherein, “gliver weight/kg body weight’ is 21, “Cells/g assembled and added 150 uL of compound-fortified plasma liver is 1.2x10, “ml incubation/number of cells in incuba to one side of each well and 150 uL of dialysis buffer to the tion” is 2.0x10, and Q, is 20 ml/min/kg. other side of each well. After 4 hours incubation at 37° C. for Supposing that hepatic metabolism is the main route of 150 rp.m. aliquots of plasma and buffer were sampled. The drug elimination, systemic exposure (AUC) after oral compound in plasma and buffer were extracted with 300 uL of 40 administration is calculated using Equation 4. acetonitrile containing internal standard compounds for AUC Dosex(1-E)/CL, Equation 4 analysis. The concentration of the compound was determined with LC/MS/MS analysis. EXAMPLES The fraction of the compound unbound was calculated by 45 the following equation: The invention is illustrated in the following non-limiting fu=1-{(plasmal-buffer)/(plasma)} examples in which, unless stated otherwise: all operations were carried out at room or ambient temperature, that is, in the wherein plasma, and buffer, are the concentrations of range of 18-25° C.; evaporation of solvent was carried out the compound in plasma and buffer, respectively. 50 using a rotary evaporator under reduced pressure with a bath temperature of up to 60°C.; reactions were monitored by thin Aqueous Solubility layer chromatography (TLC) and reaction times are given for Aqueous solubility in the mediums (a)-(c) was determined illustration only; melting points (mp) given are uncorrected by following method: (polymorphism may result in different melting points); the Whatman mini-UniPrep chambers (Clifton, N.J., USA) 55 structure and purity of all isolated compounds were assured containing more than 0.5 mg of compound and 0.5 mL of each by at least one of the following techniques: TLC (Merck silica medium were shaken overnight (over 8 hours) at room tem gel 60 F2s precoated TLC plates or Merck NH2 gel (an amine perature. All samples were filtered through a 0.45 um Poly coated silica gel) F.s. precoated TLC plates), mass spec vinylidene Difluoride (PVDF) membrane into the Whatman trometry, nuclear magnetic resonance spectra (NMR), infra mini-UniPrep plunger before analysis. The filtrates were 60 red absorption spectra (IR) or microanalysis. Yields are given assayed by HPLC. for illustrative purposes only. Workup with a cation-exchange (a) Simulated gastric fluid with no enzyme column was carried out using SCX cartridge (Varian Bond (SGN) at pH 1.2: Dissolve 2.0 g of NaCl in 7.0 mL of 10 N Elute), which was preconditioned with methanol. Flash col HCl and sufficient water to make 1000 mL.; (b) Phosphate umn chromatography was carried out using Merck silica gel buffer saline (PBS) at pH 6.5: Dissolve 6.35 g of KHPO, 65 60 (63-200 um), Wako silica gel 300HG (40-60 um), Fuji 2.84 g of NaHPO and 5.50 g of NaCl in sufficient water to Silysia NH gel (an amine coated silica gel) (30-50 um), make 1000 mL, adjusting the pH to 6.5; (c)3.94 mg of sodium Biotage KP-SIL (32-63 um) or Biotage AMINOSILICA (an US 7,718,809 B2 49 50 amine coated silica gel) (40-75 um). Preparative TLC was Example 2 carried out using Merck silica gel 60 Fasa precoated TLC plates (0.5 or 1.0 mm thickness). Low-resolution mass spec 4-(7-Fluoro-3,4-dihydro-2H-chromen-4-yl)oxy-N, tral data (EI) were obtained on an Integrity (Waters) mass N, 1,2-tetramethyl-1H-benzimidazole-6-carboxamide spectrometer. Low-resolution mass spectral data (ESI) were obtained on ZMDTM or ZQTM (Waters) and mass spectrom O eter. NMR data were determined at 270 MHz (JEOL JNM LA 270 spectrometer), 300 MHz (JEOL JNM-LA300 spec trometer) or 600 MHz (Bruker AVANCE 600 spectrometer) using deuterated chloroform (99.8% D) or dimethylsulfoxide 10 (99.9% D) as solvent unless indicated otherwise, relative to tetramethylsilane (TMS) as internal standard in parts per million (ppm); conventional abbreviations used are: S-singlet, d=doublet, t-triplet, q quartet, quint quintet, m-multiplet, bris-broad singlet, etc. IR spectra were mea 15 sured by a Fourier transform infrared spectrophotometer (Shimazu FTIR-8300). Optical rotations were measured using a JASCO DIP-370 Digital Polarimeter (Japan Spectro scopic CO., Ltd.). Chemical symbols have their usual mean ings; bp (boiling point), mp (melting point). L (liter(s)), mL (milliliter(s)), g (gram(s)), mg (milligram(s)), mol (moles), mmol (millimoles), eq. (equivalent(s)), quant. (quantitative Step 1: 7-Fluorochroman-4-ol yield), mm (millimeter(s)), min (minute(s)). To a solution of 7-fluoro-2,3-dihydro-4H-chromen-4-one 25 (6.25 g, 37.6 mmol, US 20050038032) in methanol (60 mL) Example 1 was added sodium borohydride (1.57 g., 41.4 mmol) at 0°C. The reaction mixture was stirred at the same temperature for 4-(3,4-Dihydro-2H-chromen-4-yloxy)-N.N., 1,2-tet 1 hour, and evaporated to remove methanol. The residue was ramethyl-1H-benzimidazole-6-carboxamide quenched with water, and extracted with ethyl acetate. The extract was washed with brine, dried over magnesium sulfate, 30 and concentrated in vacuum. The residue was purified by column chromatography on silica gel (hexane:ethyl acetate=3:1 as an eluent) to afford the title compound as a pale gray solid (4.70 g, 74%): 'HNMR (CDC1)8: 7.35-7.22 (m. 1H), 6.70-6.50 (m, 2H), 35 4.83-4.72 (m, 1H), 4.27 (dd, J–7.3.3.7 Hz, 2H), 2.18-1.90 (m, 3H) ppm. Step 2: 4-Chloro-7-fluorochromane 40 To a solution of 7-fluorochroman-4-ol (1.80 g, 10.7 mmol. STEP 1) and pyridine (0.35 mL) in diethyl ether (14 mL) was added thionyl chloride (3.9 mL, 53.5 mmol) at 0°C. The reaction mixture was stirred at the same temperature for 3 hours, and evaporated to remove excess thionyl chloride. The 45 residue was quenched with water, and extracted with ethyl acetate. The extract was washed with brine, dried over mag To a stirred suspension of 4-hydroxy-N,N,1,2-tetramethyl nesium sulfate, and concentrated in vacuum to afford the title 1H-benzimidazole-6-carboxamide (50 mg, 0.21 mmol. WO compound as a pale brown solid (2.0 g, quant.): 2004.054984) in N,N-dimethylformamide (2 mL) was added H NMR (CDC1) 8: 7.24 (dd, J=8.6, 5.9 Hz, 1H), 6.68 sodium hydride (60% dispersion in mineral oil, 11 mg, 0.27 6.58 (m, 1H), 6.53 (dd, J=9.9, 2.6 Hz, 1H), 5.25-5.18 (m, 1H), mmol) at room temperature. After stirring for 20 minutes, a 50 4.56-4.28 (m, 2H), 2.55-2.39 (m, 1H), 2.34-2.24 (m, 1H) solution of 4-chlorochromane (71 mg, 0.42 mmol, WO ppm. 2000078751) in N,N-dimethylformamide (1 mL) was added at room temperature. The reaction mixture was warmed to Step 3: 4-(7-Fluoro-3,4-dihydro-2H-chromen-4-yl) 70° C., and stirred for 6 hours at the same temperature. The oxy-N,N,1,2-tetramethyl-1H-benzimidazole-6-car reaction mixture was quenched with sodium hydrogencar 55 boxamide bonate aqueous Solution, and extracted with ethyl acetate (20 The title compound was prepared as a white solid in 45% mLx2). The combined extracts were washed with water and yield (0.96 g) from 4-hydroxy-N,N,1,2-tetramethyl-1H-ben brine, dried over magnesium sulfate, and concentrated in Zimidazole-6-carboxamide (1.3 g, 5.5 mmol. WO vacuum. The residue was purified by column chromatogra 60 2004.054984) and 4-chloro-7-fluorochromane (2.0 g, 11 phy on silica gel (ethylacetate:methanol-10:1 as an eluent) to mmol, STEP 2) by the same manner in STEP 1 of Example 1: afford the title compound as a white solid (40 mg, 52%): H NMR (CDC1) 8: 733-7.23 (m, 1H), 7.11 (d. J=1.3 Hz, H NMR (CDC1)8: 7.34-7.27 (m. 1H), 7.25-7.16 (m, 1H), 1H), 6.83 (d. J=1.3 Hz, 1H), 6.62-6.52 (m, 2H), 5.98 (t, J=3.3 7.11 (s, 1H), 6.91-6.80 (m, 3H), 5.99 (t, J=3.3 Hz, 1H), HZ, 1H), 4.52-4.39 (m. 1H), 4.32-4.22 (m. 1H), 3.73 (s.3H), 4.52-440 (m. 1H), 4.32-4.22 (m. 1H), 3.73 (s.3H), 3.20-2.90 65 3.11 (s, 3H), 3.02 (s, 3H), 2.62 (s, 3H), 2.42-2.30 (m, 1H), (m, 6H), 2.62 (s.3H), 2.43-2.13 (m. 2H) ppm. 2.25-2.10 (m. 1H) ppm. MS (ESI):366 (M+H), 364 (M-H). MS (ESI): 384 (M+H)". US 7,718,809 B2 51 52 Example 3 Step 1: 5,7-Difluorochroman-4-ol (-)-4-(7-Fluoro-3,4-dihydro-2H-chromen-4-yl) The title compound was prepared as a white solid in 68% oxy-N,N,1,2-tetramethv1-1H-benzimidazole-6-car yield (9.6 g) from 5,7-difluoro-2,3-dihydro-4H-chromen-4- boxamide (fraction-1) and one (14g, 77 mmol, US 20050038032) by the same manner in STEP 1 of Example 2: Example 4 'HNMR (CDC1) 8: 6.47-6.36 (m, 2H), 5.05-4.97 (m. 1H), 4.36-4.20 (m. 2H), 2.16-1.92 (m, 3H) ppm. (+)-4-(7-Fluoro-3,4-dihydro-2H-chromen-4-yl) oxy-N,N,1,2-tetramethyl-1H-benzimidazole-6-car 10 Step 2: 4-Chloro-5,7-difluorochromane boxamide (fraction-2) The title compound was prepared as a green oil in 94% The fraction-1 (337 mg) and fraction-2 (372 mg) were yield (9.0 g) from 5,7-difluorochroman-4-ol (8.6 g., 46 mmol. prepared from racemic 4-(7-fluoro-3,4-dihydro-2H STEP 1) by the same manner in STEP 2 of Example 2: chromen-4-yl)oxy-N,N,1,2-tetramethyl-1H-benzimida 15 'HNMR (CDC1)8: 6.47-6.35 (m,2H), 5.36-5.31 (m. 1H), Zole-6-carboxamide (960 mg. STEP 3 of Example 2) by 4.56-4.36 (m. 2H), 2.48-2.23 (m. 2H) ppm. HPLC as follows. Isolation Condition STEP3: 4-(5,7-Difluoro-3,4-dihydro-2H-chromen Column: CHIRALCELOJ-H (20 mmx250mm, DAICEL) 4-yl)oxy-N,N,1,2-tetramethyl-1H-benzimidazole-6- Mobile phase: n-Hexane/Ethanol/Diethylamine (85/15/ carboxamide 0.1) The title compound was prepared as a white solid in 49% Flow rate: 18.9 mL/min yield (0.12 g) from 4-hydroxy-N,N,1,2-tetramethyl-1H-ben (-)-4-(7-Fluoro-3,4-dihydro-2H-chromen-4-yl) 25 Zimidazole-6-carboxamide (0.14 g., 0.61 mmol, WO oxy-N,N,1,2-tetramethyl-1H-benzimidazole-6-car 2004.054984) and 4-chloro-5,7-difluorochromane (0.15 g. 0.73 mmol, STEP 2) by the same manner in STEP 1 of boxamide (fraction-1) Example 1: "H NMR (CDC1,) 8: 7.08 (s, 1H), 6.90 (s, 1H), 6.48-6.32 NMR: spectrum data were identical with those of the race (m. 2H), 6.08-6.00 (m. 1H), 4.55-4.42 (m. 1H), 4.35-4.24 (m, mate 30 1H), 3.71 (s, 3H), 3.09 (s, 6H), 2.60 (s, 3H), 2.45-2.33 (m, optical rotation: C-129.4° (C=1.11, Methanol) 1H), 2.17-1.97 (m. 1H) ppm: retention time: 16.7 min MS (ESI): 402 (M+H)". (+)-4-(7-Fluoro-3,4-dihydro-2H-chromen-4-yl) Example 6 oxy-N,N,1,2-tetramethyl-1H-benzimidazole-6-car 35 boxamide (fraction-2) (-)-4-(5,7-Difluoro-3,4-dihydro-2H-chromen-4-yl) oxy-N,N,1,2-tetramethyl-1H-benzimidazole-6-car NMR: spectrum data were identical with those of the race boxamide and mate optical rotation: C-124.0° (C=1.07, Methanol) 40 Example 7 retention time: 21.0 min (+)-4-(5,7-Difluoro-3,4-dihydro-2H-chromen-4-yl) Example 5 oxy-N,N,1,2-tetramethyl-1H-benzimidazole-6-car boxamide 4-(5,7-Difluoro-3,4-dihydro-2H-chromen-4-yl) 45 oxy-N,N,1,2-tetramethyl-1H-benzimidazole-6-car The fraction-1 (1.13 g) and fraction-2 (1.09 g) were pre boxamide pared from racemic 4-(5,7-difluoro-3,4-dihydro-2H chromen-4-yl)oxy-N,N,1,2-tetramethyl-1H-benzimida 50 Zole-6-carboxamide (3.05 g, STEP3 of Example 5) by HPLC as follows. O Isolation Condition N N Column: CHIRALPAK AD-H (20 mmx250 mm, DAICEL) 2 55 Mobile phase: n-Hexane/Ethanol/Diethylamine (80/20/ N 0.1) O Flow rate: 18.9 mL/min

60 (-)-4-(5,7-Difluoro-3,4-dihydro-2H-chromen-4-yl) O F oxy-N,N,1,2-tetramethyl-1H-benzimidazole-6-car boxamide (fraction-1) NMR: spectrum data were identical with those of the race 65 mate optical rotation: C-119.3° (C=1.00, Methanol) retention time: 9.4 min

US 7,718,809 B2 55 56 Step 1: 4-Chloro-3,4-dihydro-1H-isochromene reaction mixture was quenched with water and extracted with ethyl acetate (250 mLx2). The combined extracts were The title compound was prepared as a yellow oil in 99% washed with sodium hydrogencarbonate aqueous solution, yield (6.6 g) from 3,4-dihydro-1H-isochromen-4-ol (5.9 g, 39 ammonium chloride aqueous Solution and brine, dried over mmol, WO 2004024081) by the same manner in STEP 2 of 5 Sodium Sulfate, and concentrated in vacuum. The resulting Example 2: solid was suspended with ethyl acetate and diethyl ether, and 'HNMR (CDC1,)8: 7.54-7.42(m, 1H), 7.35-7.22 (m, 2H), collected by filtration to give the title compound as a white 7.08-6.97 (m, 1H), 5.18-5.08 (m, 1H), 4.91 (d. J=15.0 Hz, solid (2.95 g, 61%): 1H), 4.78 (d. J=15.2 Hz, 1H), 4.27-4.08 (m, 2H) ppm. 10 H NMR (CDC1) 8: 7.75 (s, 1H), 7.55 (s, 1H), 7.36 (dd, J=7.9, 1.3 Hz, 1H), 7.27-7.15 (m, 1H), 6.90-6.80 (m, 2H), Step 2: 4-(3,4-Dihydro-1H-isochromen-4-yloxy)-N, 5.96-5.90 (m. 1H), 4.52-440 (m, 1H), 4.32-4.25 (m. 1H), N, 1,2-tetramethyl-1H-benzimidazole-6-carboxamide 3.95 (s, 3H), 3.77 (s, 3H), 2.63 (s, 3H), 2.42-2.20 (m, 2H) ppm, The title compound was prepared as a white solid in 12% 15 yield (38 mg) from 4-hydroxy-N,N,1,2-tetramethyl-1H-ben MS (ESI): 353 (M+H)". Zimidazole-6-carboxamide (0.20 g, 0.85 mmol, WO 2004.054984) and 4-chloro-3,4-dihydro-1H-isochromene Step 2: 4-(3,4-Dihydro-2H-chromen-4-yloxy)-1,2- (0.20 g, 1.2 mmol, STEP 1) by the same manner in STEP 1 of dimethyl-1H-benzimidazole-6-carboxylic acid Example 1: "H NMR (CDC1) 8: 7.50 (d. J=7.3 Hz, 1H), 7.34-7.17 (m, To a stirred solution of methyl 4-(3,4-dihydro-2H 2H), 7.14 (s, 1H), 7.06 (d. J–7.3 Hz, 1H), 6.88 (s, 1H), chromen-4-yloxy)-1,2-dimethyl-1H-benzimidazole-6-car 6.09-6.00 (m. 1H), 4.91 (d. J=15.1 Hz, 1H), 4.77 (d. J=15.1 boxylate (1.0g, 2.8 mmol, STEP 1) in methanol (20 mol) and HZ, 1H), 4.32 (dd, J=12.5, 4.0 Hz, 1 H), 4.11 (dd, J=12.5.3.3 25 tetrahydrofuran (30 mL) was added 4M lithium hydroxide HZ, 1H), 3.72 (s.3H), 3.09 (s.3H), 3.02 (s.3H), 2.61 (s.3H) aqueous solution (4 mL, 16 mmol) at room temperature. After ppm, stirring for 5 hours at 60° C., the reaction mixture was con MS (ESI):366 (M+H), 364 (M-H). centrated in vacuum. The residue was dissolved with water Example 11 and acidified (pH-3) with 2M hydrochloric acid. The result 30 ing precipitate was filtrated, and dried in vacuum to afford the 4-(3,4-Dihydro-2H-chromen-4-yloxy)-N-(2-hy title compound as a white solid (0.85g. 89%): droxyethyl)-N,1,2-trimethyl-1H-benzimidazole-6- H NMR (CDC1) 8: 7.81 (s, 1H), 7.62 (s, 1H), 7.38-7.34 carboxamide (m. 1H), 7.27-7.18 (m, 1H), 6.90-6.83 (m, 2H), 5.93-5.90 (m, 35 1H), 4.53-4.50 (m. 1H), 4.32-4.26 (m. 1H), 3.80 (s.3H), 2.68 (s, 3H), 2.40-2.10 (m. 2H) ppm ( OH was not observed); O MS (ESI): 339 (M+H)",337 (M-H). / HO N-1SN N Step 3: 4-(3,4-Dihydro-2H-chromen-4-yloxy)-N-(2- 40 hydroxyethyl)-N,1,2-trimethyl-1H-benzimidazole-6- N carboxamide cr;O To a stirred mixture of 4-(3,4-dihydro-2H-chromen-4- 45 yloxy)-1,2-dimethyl-1H-benzimidazole-6-carboxylic acid (70 mg, 0.21 mmol, STEP 2) and 2-(methylamino)ethanol (31 mg, 0.41 mmol) in dimethylformamide (2 mL) were added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) (67 mg, 0.35 mmol) and 1-hydroxy 50 benzotriazole hydrate (HOBt) (60 mg, 0.41 mmol) at room temperature. After stirring at room temperature for 18 hours, Step 1: Methyl 4-(3,4-dihydro-2H-chromen-4- the reaction mixture was quenched with sodium hydrogen yloxy)-1,2-dimethyl-1H-benzimidazole-6-carboxy carbonate aqueous solution (5 mL), and extracted with ethyl late 55 acetate (50 mLx2). The combined extracts were washed with brine, dried over sodium sulfate, and concentrated in vacuum. To a suspension of sodium hydride (60% dispersion in The residue was purified by column chromatography on silica mineral oil, 0.71 g, 18 mmol) in N,N-dimethylformamide (40 gel (ethyl acetate:methanol=10:1 as an eluent). The residue mL) was added a Suspension of methyl 4-hydroxy-1,2-dim was suspended with diethyl ether, and collected by filtration ethyl-1H-benzimidazole-6-carboxylate (3.0 g, 14 mmol. WO 60 to afford the title compound as a white solid (24 mg, 24%): 2004.054984) in N,N-dimethylformamide (40 mL) dropwise under nitrogen atmosphere at room temperature. After stir H NMR (CDC1) 8: 7.30-7.16 (m,3H), 6.92-6.82 (m,3H), ring for 20 minutes, a solution of 4-chlorochromane (4.6 g. 27 6.02-5.97 (m. 1H), 4.50-4.43 (m, 1H), 4.33-4.23 (m. 1H), mmol, WO 2000078751) in N,N-dimethylformamide (10 3.95-3.88 (m, 2H), 3.75-3.50 (m, 2H), 3.73 (s, 3H), 3.07 (s, mL) was added at room temperature. The reaction mixture 65 3H), 2.62 (s.3H), 2.40-2.34 (m. 1H), 2.28-2.18 (m. 1H) ppm was warmed to 70° C. for 2.5 hours, cooled to room tempera (—OH was not observed): ture, and stirred for 18 hours at the same temperature. The MS (ESI): 396 (M+H)".

US 7,718,809 B2 63 64 (+)-6-(AZetidin-1-ylcarbonyl)-4-(5,7-difluoro-3,4- Step 1: 3-Methylazetidin-3-ol hydrochloride dihydro-2H-chromen-4-yl)oxyl-1,2-dimethyl-1H benzimidazole (fraction-2) 1-(Diphenylmethyl)-3-methylazetidin-3-ol (0.48 g, 1.9 mmol) and 10% palladium on activated carbon (0.20 g) in NMR: spectrum data were identical with those of the race- 5 methanol (4 mL) was stirred under hydrogen gas (4 atmo mate spheres) for 10 hours. The resulted mixture was filtered through a pad of Celite. 4M hydrogen chloride in dioxane (1 optical rotation: O’=+119.8° (C=0.49, Methanol) mL) was added to the filtrate, and the mixture was concen retention time: 26 min trated in vacuum to afford the title compound as crude oil 10 (0.38 g). Example 25 Step2: 1-(4-(5,7-Difluoro-3,4-dihydro-2H 4-(5,7-Difluoro-3,4-dihydro-2H-chromen-4-yl) chromen-4-yl)oxyl-1,2-dimethyl-1H-benzimidazol oxy-1,2-dimethyl-6-(pyrrolidin-1-ylcarbonyl)-1H 6-y1}carbonyl)-3-methylazetidin-3-ol benzimidazole 15 The title compound was prepared as a colorless amorphous in 76% yield (71 mg) from 4-(5,7-difluoro-3,4-dihydro-2H chromen-4-yl)oxyl-1,2-dimethyl-1H-benzimidazole-6-car boxylic acid (80 mg, 0.21 mmol, STEP 2 of Example 21) and 3-methylazetidin-3-ol hydrochloride (crude 135 mg, STEP 1) by the same manner in STEP 3 of Example 11. H NMR (CDC1) 8: 7.25-7.15 (m. 1H), 7.11 (brs, 1H), 6.48-6.33 (m, 2H), 6.05-5.98 (m, 1H), 4.53-4.41 (m. 1H), 4.35-4.13 (m, 5H), 3.67 (s.3H), 2.58 (s, 3H), 2.43-2.32 (m, 25 1H), 2.16-2.02 (m. 1H), 1.56 (s, 3H) ppm ( OH was not observed.); MS (ESI): 444 (M+H)". All publications, including but not limited to, issued pat ents, patent applications, and journal articles, cited in this application are each herein incorporated by reference in their entirety. Although the invention has been described above with reference to the disclosed embodiments, those skilled in the The title compound was prepared as a colorless amorphous art will readily appreciate that the specific experiments in 39% yield (45 mg) from 4-(5,7-difluoro-3,4-dihydro-2H detailed are only illustrative of the invention. It should be chromen-4-yl)oxyl-1,2-dimethyl-1H-benzimidazole-6-car understood that various modifications can be made without boxylic acid (100 mg, 0.27 mmol, STEP 2 of Example 19) departing from the spirit of the invention. Accordingly, the and pyrrolidine (38 mg, 0.53 mmol) by the same manner in invention is limited only by the following claims. STEP 3 of Example 11. H NMR (CDC1) 8: 7.20 (s, 1H), 7.01 (s, 1H), 6.48-6.32 40 The invention claimed is: (m. 2H), 6.07-6.00 (m, 1H), 4.55-4.42 (m, 1H), 4.35-4.25 (m, 1. A compound of the formula (I): 1H), 3.78-3.63 (m, 2H), 3.72 (s.3H), 3.55-3.42 (m, 2H), 2.60 (s, 3H), 2.45-2.34 (m. 1H), 2.15-1.80 (m, 5H) ppm. (I) MS (ESI): 428 (M+H)". 45 Example 26 1-(4-(5,7-Difluoro-3,4-dihydro-2H-chromen-4-yl) oxy-1,2-dimethyl-1H-benzimidazol-6-y1}carbonyl)- 3-methylazetidin-3-ol

O

N N HO 2 N

O 60 O F

X is an oxygen atom or NH; 65 R" and Rare each independently C-C alkyl each being unsubstituted or substituted with 1 to 2 substituents independently selected from hydroxy or C-C alkoxy; US 7,718,809 B2 65 66 Rand Rare each independently hydrogen, C-C alkyl, R. R. and Rare each hydrogen; or C-C, cycloalkyl, said C-C alkyl and said C-C, or a pharmaceutically acceptable salt thereof. cycloalkyl being unsubstituted or substituted with 1 to 3 3. The compound of claim 1, wherein: Substituents independently selected from halogen, -A-B - is —CH2—O—; hydroxy, C-C alkoxy, or C-C, cycloalkyl; or R and X is oxygen; R' taken together with the nitrogen atom to which they R" and Rare each methyl: are attached form a 4 to 6 membered heterocyclic group RandR are each methyl; or RandR' taken together with being unsubstituted or substituted with 1 to 3 substitu the nitrogen atom to which they are attached form an ents selected from hydroxy, C-C alkyl, C-C acyl, or aZetidin-1-yl, pyrrolidin-1-yl, 3-hydroxy-pyrrolidin-1- hydroxy-C-C alkyl; 10 yl, or 4-acetyl-piperazin-1-yl group; R. R. R. and Rare each independently hydrogen, halo RandR are independently hydrogen, halogen, or methyl; gen, hydroxy, C-C alkyl, or C-C alkoxy; and and R is hydrogen, hydroxy, or C1-C4 alkoxy; or R. RandR are each a hydrogen atom; a pharmaceutically acceptable salt thereof. or a pharmaceutically acceptable salt thereof. 15 4. The compound of claim 1, which is: 2. The compound of claim 1, wherein: 4-(7-Fluoro-3,4-dihydro-2H-chromen-4-yl)oxy-N,N.1, X is an oxygen atom; 2-tetramethyl-1H-benzimidazole-6-carboxamide: R and Rare each independently hydrogen, C-C alkyl, 4-(5,7-Difluoro-3,4-dihydro-2H-chromen-4-yl)oxy-N, or C-C cycloalkyl, said C-C alkyl and said C-C, N, 1,2-tetramethyl-1H-benzimidazole-6-carboxamide: cycloalkyl being unsubstituted or substituted with 1 to 3 O Substituents independently selected from hydrogen, N.N.1.2-Tetramethyl-4-(5-methyl-3,4-dihydro-2H hydroxyl, C-C alkoxy, or C-C, cycloalkyl; or R and chromen-4-yl)oxy-1H-benzimidazole-6-carboxamide: R' taken together with the nitrogen atom to which they or a pharmaceutically acceptable salt thereof. are attached form an aZetidinyl, pyrrolidinyl, or piper 5. A pharmaceutical composition comprising the com azinyl group, said aZetidinyl, pyrrolidinyl group, and 25 pound or the pharmaceutically acceptable salt thereof of piperazinyl groups being unsubstituted or Substituted claim 1, and at least one pharmaceutically acceptable carrier. with 1 to 3 substituents selected from hydroxyl, C-C, alkyl, C-C acyl, or hydroxyl-C-C alkyl, and k k k k k