IL212727A - Modulators of atp-binding cassette transporters, pharmaceutical compositions comprising them and use thereof in the preparation of medicaments for the treatment of diseases - Google Patents

Modulators of atp-binding cassette transporters, pharmaceutical compositions comprising them and use thereof in the preparation of medicaments for the treatment of diseases

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IL212727A
IL212727A IL212727A IL21272711A IL212727A IL 212727 A IL212727 A IL 212727A IL 212727 A IL212727 A IL 212727A IL 21272711 A IL21272711 A IL 21272711A IL 212727 A IL212727 A IL 212727A
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Vertex Pharma
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Description

ATP-mtinp D^irm ποορ ^iu D^.EJND MODULATORS OF ATP-BINDING CASSETTE TRANSPORTERS MODULATORS OF ATP-BINDING CASSETTE TRANSPORTERS CROSS REFERNCE TO RELATED APPLICATIONS
[0001] 'ITiis application claims priority to U.S. Provisional Application Serial No. 61/1 2, 152, which was filed November 6, 2008. The entire contents of U.S. Provisional Application Serial No. 61 /1 12, 152 is incorporated herein by reference.
TECHNICAL FIELD OF THE INVEN TION
[0002] The present invention relates to modulators of ATP-Binding Cassette ("ABC") transporters or fragments thereof, including Cystic Fibrosis Transmembrane Conductance Regulator ("CFTR"), compositions thereof and methods therewith. The present invention also relates to methods of treating ABC transporter mediated diseases using such modulators.
BACKGROUND OF THE INVENTION
[0003] ABC transporters are a family of membrane transporter proteins that regulate the transport of a wide variety of pharmacological agents, potentially toxic drugs, and xenobiotics, as well as anions. ABC transporters are homologous membrane proteins that bind and use cellular adenosine triphosphate (ATP) for their specific activities. Some of these transporters were discovered as multidrug resistance proteins (like the MDR l -P glycoprotein, or the multidrug resistance protein, MRPI ), defending malignant cancer cells against chcmothcrapeutic agents. To date, 48 ABC Transporters have been identified and grouped into 7 families based on their sequence identity and function.
[0004] ABC transporters regulate a variety of important physiological roles within the body and provide defense against harmful environmental compounds. Because of this, they represent important potential drug targets for the treatment of diseases associated with defects in the transporter, prevention of drug transport out of the target cell, and intervention in other diseases in which modulation of ABC transporter activity may be beneficial.
[0005] One member of the ABC transporter family commonly associated with disease is the cAMP/ATP-medialed anion channel, CFTR. CFTR is expressed in a variety of cells types, including absorptive and secretory epithelia cells, where it regulates anion flux across the membrane, as well as the activity of other ion channels and proteins. In epithelia cells, normal functioning of CFTR is critical for the maintenance of electrolyte transport throughout the body, including respiratory and digestive tissue. CFTR is composed of approximately 1480 amino acids that encode a protein made up of a tandem repeat of transmembrane domains, each containing six transmembrane helices and a nucleotide binding domain. The two transmembrane domains are linked by a large, polar, regulatory (R)-domain with multiple phosphorylation sites that regulate channel activity and cellular trafficking.
[0006] The gene encoding CFTR has been identified and sequenced (See Gregory, R. J. et al. ( 1990) Nature 347:382-386; Rich, D. P. et al. ( 1990) Nature 347:358-362), (Riordan, J. R. et al. (1989) Science 245: 1066-1073). Λ defect in this gene causes mutations in CFTR resulting in Cystic Fibrosis ("CF"), the most common fatal genetic disease in humans. Cystic Fibrosis affects approximately one in every 2,500 infants in the United States. Within the general United Slates population, up to 10 million people carry a single copy of the defective gene without apparent ill effects. In contrast, individuals with two copies of the CF associated gene suffer from the debilitating and fatal effects of CF, including chronic lung disease.
[0007] In patients with cystic fibrosis, mutations in CFFR endogenously expressed in respiratory epithelia leads to reduced apical anion secretion causing an imbalance in ion and fluid transport. The resulting decrease in anion transport contributes to enhanced mucus accumulation in the lung and the accompanying microbial infections that ultimately cause death in CF patients. In addition to respiratory disease, CF patients typically suffer from gastrointestinal problems and pancreatic insufficiency that, if left untreated, results in death. In addition, the majority of males with cystic fibrosis are infertile and fertility is decreased among females with cystic fibrosis. In contrast to the severe effects of two copies of the CF associated gene, individuals with a single copy of the CF associated gene exhibit increased resistance to cholera and to dehydration resulting from diarrhea - perhaps explaining the relatively high frequency of the CF' gene within the population.
[0008] Sequence analysis of the CFrR gene of CF chromosomes has revealed a variety of disease causing mutations (Cutting, G. R. et al. (1990) Nature 346:366-369; Dean, M. et al. (1990) Cell 61 :863:870; and Kerem, 13-S. et al. (1989) Science 245: 1073- 1080; Kerem, B-S el al. (1990) Proc. Natl. Acad. Sci. USA 87:8447-8451). To date, > 1000 disease causing mutations in the CF genc have been identified (http://www.genel.sickkids.on.ca/cflr/). The most prevalent mutation is a deletion of phenylalanine at position 508 of the CF R amino acid sequence, and is commonly referred to as ΔΡ'508-CFTR. This mutation occurs in approximately 70% of the cases of cystic fibrosis and is associated with a severe disease.
[0009] The deletion of residue 508 in AI7508-CFFR prevents the nascent protein from folding correctly. This results in the inability of the mutant protein to exit the ER, and traffic to the plasma membrane. As a result, (he number of channels present in the membrane is far less than observed in cells expressing wild-type CI R. In addition to impaired trafficking, the mutation results in defective channel gating. 'I'ogcther, the reduced number of channels in the membrane and the defective gating lead to reduced anion transport across epithelia leading to defective ion and fluid transport. (Quinton, P. M. (1990), FASEB J. 4: 2709-2727). Studies have shown, however, that the reduced numbers of AF508-CFrR in the membrane are functional, albeit less than wild-type CFl'R. (Dalemans et al. ( 1991 ), Nature Lond. 354: 526-528; Denning et al., supra; Pasyk and Foskett (1995), J. Cell. Biochem. 270: 12347-50). In addition to AF508-CI TR, other disease causing mutations in CFl'R that result in defective trafficking, synthesis, and/or channel gating could be up- or down-regulated to alter anion secretion and modify disease progression and/or severity.
[0010] Although CFTR transports a variety of molecules in addition to anions, it is clear that this role (the transport of anions) represents one element in an important mechanism of transporting ions and water across the epithelium. The other elements include the epithelial Na+ channel, ENaC, Na+/2C17 + co- transporter, Na+- +-ATPase pump and the basolateral membrane K+ channels, that are responsible for the uptake of chloride into the cell.
[0011] These elements work together to achieve directional transport across the epithelium via their selective expression and localization within the cell. Chloride absorption takes place by the coordinated activity of ENaC and CFl'R present on the apical membrane and the Na+-K+-ATPase pump and CI- channels expressed on the basolateral surface of the cell. Secondary active transport of chloride from the luminal side leads to the accumulation of intracellular chloride, which can then passively leave the cell via CI" channels, resulting in a vectorial transport. Arrangement of Na+/2C17 + co-transporter, Na+-K+-ATPase pump and the basolateral membrane + channels on the basolateral surface and CFl'R on the luminal side coordinate the secretion of chloride via CFl'R on the luminal side. Because water is probably never actively transported itself, its flow across epithelia depends on tiny transepithelial osmotic gradients generated by the bulk flow of sodium and chloride.
[0012] In addition to Cystic Fibrosis, modulation of CFTR activity may be beneficial for other diseases not directly caused by mutations in CFTR, such as secretory diseases and other protein folding diseases mediated by CFTR. These include, but are not limited to, chronic obstructive pulmonary disease (COPD), dry eye disease, and Sjogren's Syndrome.
[0013] COPD is characterized by airflow limitation that is progressive and not fully reversible. The airflow limitation is due to mucus hypersecretion, emphysema, and bronchiolitis. Activators of mutant or wild-type H R offer a potential treatment of mucus hypersecretion and impaired mucociliary clearance that is common in COPD. Specifically, increasing anion secretion across CFFR may facilitate fluid transport into the airway surface liquid to hydrate the mucus and optimized perici!iary fluid viscosity. This would lead to enhanced mucociliary clearance and a reduction in the symptoms associated with COPD. Dry eye disease is characterized by a decrease in tear aqueous production and abnormal tear film lipid, protein and mucin profiles. There are many causes of dry eye, some of which include age, Lasik eye surgery, arthritis, medications, chemical/thermal burns, allergies, and diseases, such as Cystic Fibrosis and SjOgrens's syndrome. Increasing anion secretion via CFl'R would enhance fluid transport from the corneal endothelial cells and secretory glands surrounding the eye to increase corneal hydration. This would help to alleviate the symptoms associated with dry eye disease. SjOgrens's syndrome is an autoimmune disease in which the immune system attacks moisture-producing glands throughout the body, including the eye, mouth, skin, respiratory tissue, liver, vagina, and gut. Symptoms, include, dry eye, mouth, and vagina, as well as lung disease. The disease is also associated with rheumatoid arthritis, systemic lupus, systemic sclerosis, and polymypositis/dermatomyositis. Defective protein trafficking is believed to cause the disease, for which treatment options are limited.
Modulalors of CFTR activity may hydrate the various organs afflicted by the disease and help to elevate the associated symptoms.
[0014] As discussed above, it is believed that the deletion of residue 508 in AF508-CFTR prevents the nascent protein from folding correctly, resulting in the inability of this mutant protein to exit the ER, and traffic to the plasma membrane. As a result, insufficient amounts of the mature protein are present at the plasma membrane and chloride transport within epithelial tissues is significantly reduced. In fact, this cellular phenomenon of defective BR processing of ABC transporters by the ER machinery has been shown to be the underlying basis not only for CF disease, but for a wide range of other isolated and inherited diseases. The two ways that the ER machinery can malfunction is either by loss of coupling to ER export of the proteins leading to degradation, or by the ER accumulation of these defeclive/misfolded proteins [Aridor M, et al., Nature Med., 5(7), pp 745- 751 ( 1999); Shastry, B.S., el al., Neurochem. International, 43, pp 1 -7 (2003); Rutishauser, J., et al., Swiss Med Wkly, 132, pp 211-222 (2002); Morello, JP et al., TIPS, 21, pp. 466- 469 (2000); Bross P., et al., Human Mut., 14, pp. 186- 1 8 ( 1999) |. The diseases associated with the first class of ER malfunction are Cystic fibrosis (due to misfolded AF508-CFfR as discussed above), Hereditary emphysema (due to a I -antitrypsin; non Piz variants), Hereditary hemochromatosis, Coagu!ation-Fibrinolysis deficiencies, such as Protein C deficiency, Type 1 hereditary angioedema, Lipid processing deficiencies, such as Familial hypercholesterolemia, Type 1 chylomicronemia, Abetalipoproteinemia, Lysosomal storage diseases, such as 1-cell disease/Pseudo- Hurler, Mucopolysaccharidoses (due to Lysosomal processing enzymes), Sandhof/Tay-Sachs (due to β-Hexosaminidase), Crigler-Najjar type II (due to UDP-glueuronyl-sialyc-transferase), Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus (due to Insulin receptor), Laron dwarfism (due to Growth hormone receptor), Myleoperoxidase deficiency, Primary hypoparathyroidism (due to Preproparathyroid hormone), Melanoma (due to Tyrosinase). The diseases associated with the latter class of ER malfunction arc Glycanosis CDG type 1 , Hereditary emphysema (due to a 1 -Antitrypsin (PiZ variant), Congenital hyperthyroidism, Osteogenesis imperfecta (due to Type 1, 11, IV procollagen), Hereditary hypofibrinogeneniia (due to Fibrinogen), ACT deficiency (due to al-Antichymotrypsin), Diabetes insipidus (DI), Neurophyseal DI (due to Vasopvessin hormone/V2-receptor), Neprogenic DI (due to Aquaporin II), Charcot-Marie Tooth syndrome (due to Peripheral myelin protein 22), Perlizaeus-Merzbachcr disease, neurodegenerative diseases such as Alzheimer's disease ( due to βΑΡΡ and presenilins), Parkinson's disease, Amyotrophic lateral sclerosis, Progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders asuch as Huntington, Spinocerebullar ataxia type I, Spinal and bulbar muscular atrophy, Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as Spongiform encephalopathies, such as Hereditary Creulzfeldt- Jakob disease (due to Prion protein processing defect), Fabry disease (due to lysosomal a-galactosidase A) and Straussler-Scheinker syndrome (due to Prp processing defect).
[0015] In addition to up-regulation of CFTR activity, reducing anion secretion by CFPR modulators may be beneficial for the treatment of secretory diarrheas, in which epithelial water transport is dramatically increased as a result of secretagogue activated chloride transport. The mechanism involves elevation of cAMP and stimulation of CFFR.
[0016] Although there are numerous causes of diarrhea, the major consequences of diarrheal diseases, resulting from excessive chloride transport are common to all, and include dehydration, acidosis, impaired growth and death.
[0017] Acute and chronic diarrheas represent a major medical problem in many areas of the world. Diarrhea is both a significant factor in malnutrition and the leading cause of death (5,000,000 deaths/year) in children less than five years old.
[0018] Secretory diarrheas are also a dangerous condition in patients of acquired immunodeficiency syndrome (AIDS) and chronic inflammatory bowel disease (IBD). 1 6 million travelers to developing countries from industrialized nations every year develop diarrhea, with the severity and number of cases of diarrhea varying depending on the country and area of travel. [0019J Diarrhea in barn animals and pels such as cows, pigs and horses, sheep, goats, cats and dogs, also known as scours, is a major cause of death in these animals. Diarrhea can result from any major transition, such as weaning or physical movement, as well as in response to a variety of bacterial or viral infections and generally occurs within the first few hours of the animal's life.
[0020] The most common diarrhea causing bacteria is enteroloxogenic E-coli (ETEC) having the 99 pilus antigen. Common viral causes of diarrhea include rotavirus and coronavirus. Other infectious agents include Cryptosporidium, giardia lamblia, and salmonella, among others.
[0021] Symptoms of rotaviral infection include excretion of watery feces, dehydration and weakness. Coronavirus causes a more severe illness in the newborn animals, and has a higher mortality rate than rotaviral infection. Often, however, a young animal may be infected with more than one virus or with a combination of viral and bacterial microorganisms at one time. This dramatically increases the severity of the disease.
[0022] Accordingly, there is a need for modulators of an ABC transporter activity, and compositions thereof, that can be used to modulate the activity of the ABC transporter in the cell membrane of a mammal.
[0023] There is a need for methods of treating ABC transporter mediated diseases using such modulators of ABC transporter activity.
[0024] There is a need for methods of modulating an ABC transporter activity in an ex vivo cell membrane of a mammal.
[0025] There is a need for modulators of CFJ'R activity that can be used to modulate the activity of CFTR in the cell membrane of a mammal.
[0026] There is a need for methods of treating CFITi-mediated diseases using such modulators of C1 R activity.
[0027] l nere is a need for methods of modulating CI R activity in an ex vivo cell membrane of a mammal.
[0028] SUMMARY OF THE INVENTION
[0029] It has now been found thut compounds of this invention, and phannaceutically acceptable compositions thereof, are useful as modulators of ABC transporter activity, particularly CTFR activity. These compounds have the general formula I: or a phannaceutically acceptable salt thereof, wherein R1 t R2, ring A, ring B, and n are defined below.
It has also now been found that compounds of this invention, and phan aceutically acceptable compositions thereof, are useful as modulators of ABC transporter activity. Ihese compounds have the general formula II : II or a phannaceutically acceptable salt thereof, wherein R, R], R2, Rj, R4, and R5 are defined below.
[0030] ITiese compounds and pharmaceutically acceptable compositions are useful for treating or lessening the severity of a variety of diseases, disorders, or conditions, including, but not limited to, cystic fibrosis, hereditary emphysema, hereditary hemochromatosis, coagulation- fibrinolysi deficiencies, such as protein C deficiency, Type I hereditary angioedema, lipid processing deficiencies, such as familial hypercholesterolemia, Type I chylomicroncmia, abetalipoproieincmia, lysosomal storage diseases, such as I-ccll disease/pseudo-Hurler, mucopolysaccharidoses, Sandhol/l ay-Sachs, Crigler-Najjar type II, polyendocrinopathy/hyperinsulemia, diabetes mellitus, laron dwarfism, myleoperoxidase deficiency, primary hypoparathyroidism, melanoma, glycanosis CDG type 1 , hereditary emphysema, congenital hyperthyroidism, osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency, diabetes insipidus, neurophysiol, nephrogenic, Charcot- Marie Tooth syndrome, Perlizaeus-Merzbachcr disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders asuch as Huntington, spinocerehullar ataxia type I, spinal and bulbar muscular atrophy, dentatorubal pallidoluysian, and myotonic dystrophy, as well as spongi form encephalopathies, such as hereditary Creutzfeldt-Jakob disease, Fabry disease, Straussler-Schcinker syndrome, COPD, dry-eye disease, and Sjogren's disease.
[0031] DETAILED DESCRIPTION OF THE INVENTION
[0032] 1. DEFINITIONS
[0033] As used herein, the following definitions shall apply unless otherwise indicated.
[0034] The term "ABC-transporter" as used herein means an AI C -transporter protein or a fragment thereof comprising at least one binding domain, wherein said protein or fragment thereof is present in vivo or in vitro. The term "binding domain" as used herein means a domain on the ABC-transporter that can bind to a modulator. See, e.g., Hwang, T. C. et al., J. Gen. Physiol. (1998): 1 1 1(3), 477-90.
[0035] The term "CFTR" as used herein means cystic fibrosis transmembrane conductance regulator or a mutation thereof capable of regulator activity, including, but not limited to, AF508 CFTR and G551 D CFTR (sec, e.g., http://www.genet.sickkids.on.ca cftr/, for CI R mutations).
[0036] The term "modulating" as used herein means increasing or decreasing, e.g. activity, by a measurable amount. Compounds that modulate ABC Transporter activity, such as CFfR activity, by increasing the activity of the ABC Transporter, e.g., a CFI'R anion channel, are called agonists. Compounds that modulate ABC- Transporter activity, such as CFI'R activity, by decreasing the activity of the ABC Transporter, e.g., CFFR anion channel, are called antagonists. An agonist interacts with an ABC Transporter, such as CFI'R anion channel, to increase the ability of the receptor to transduce an intracellular signal in response lo endogenous ligand binding. An antagonist interacts with an ABC Transporter, such as CFTR, and competes with the endogenous ligand(s) or substrate(s) for binding sile(s) on the receptor lo decrease the ability of Ihe receptor to transduce an intracellular signal in response to endogenous ligand binding.
[0037] The phrase "treating or reducing the severity of an ABC Transporter mediated disease" refers both to treatments for diseases that are directly caused by ABC Transporter and/or CFrR activities and alleviation of symptoms of diseases not directly caused by ABC Transporter and/or CFTR anion channel activities. Examples of diseases whose symptoms may be affected by ABC Transporter and/or CFTR activity include, but are not limited to, Cystic fibrosis, Hereditary emphysema, Hereditary hemochromalosis, Coagulalion-Fibrinolysis deficiencies, such as Protein C deficiency, Type 1 hereditary angioedema, Lipid processing deficiencies, such as Familial hypercholesterolemia, Type 1 chylomicronemia, Abctalipoprotcincmia, Lysosomal storage diseases, such as I-ccll discasc Pseudo- Hurler, Mucopolysaccharidoses, Sandhof' Tay-Sachs, Crigler-Najjar type II, Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus, Laron dwarfism, Myleoperoxidase deficiency, Primary hypoparathyroidism, Melanoma, Glycanosis CDG type I , Hereditary emphysema, Congenital hyperthyroidism, Osteogenesis imperfecta, Hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (Dl), Neurophysiol DI, Nephrogenic Dl, Charcoi-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, Progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders such as Huntington, Spinocerebullar ataxia type I, Spinal and bulbar muscular atrophy, Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as Spongiform encephalopathies, such as Hereditary Creutzfeldt-Jakob disease, Fabry disease, Straussler-Scheinker syndrome, COPD, dry-eye disease, and Sjogren's disease.
[0038] For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed. Additionally, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausolito: 1999, and "March's Advanced Organic Chemistry", 5th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001 , the entire contents of which are hereby incorporated by reference.
[0039] As described herein, compounds of the invention may optionally be substituted with one or more substituents, such as are illustrated generally above, or as exemplified by particular classes, subclasses, and species of the invention.
[0040] As used herein the term "aliphatic" encompasses the temis alkyl, alkenyl, alkynyl, each of which being optionally substituted as set forth below.
[0041] As used herein, an "alkyl" group refers to a saturated aliphatic hydrocarbon group containing 1 -12 (e.g., 1 -8, 1 -6, or I -4) carbon atoms. Λη alkyl group can be straight or branched. Examples of" alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-heptyl, or 2-ethylhexyl. An alkyl group can be substituted (i.e., optionally substituted) with one or more substituenLs such as halo, phospho, cycloaliphalic [e.g., cycloalkyl or cycloalkenyl], heterocycloaliphatic |e.g., heterocycloalky! or heterocycloalkenyl], aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl [e.g., (aliphatic)carbonyl, (cycloaliphatic)carbonyl, or (heterocycloaliphatic)carbonyl], nitro, cyano, amido [e.g., (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (hetcrocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylamino alkylaminocarbonyl, cycloalkylaminocarbonyl, hctcrocycloalkylaminocarbonyl, arylaminocarbonyl, or heteroarylaminocarbonyl] , ami no |e.g. , aliphaticamino, cycloaliphaticamino, or heterocycloaliphaticaminoj, sulfonyl je.g. , aliphatic-S02-j, sulfinyl, sulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, carboxy, carbamoyl, cycloaliphalicoxy, heterocycloaliphaticoxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaryl alkoxy, alkoxycarbonyl, alkylcarbonyloxy, or hydroxy. Without limitation, some examples of substituted alkyls include carboxyalkyl (such as MOOC-alkyl, alkoxycarbonylalkyl, and alkylearbonyloxyalkyl), cyanoalkyl, hydroxyalkyl, alkoxyalkyl, acylalkyl, aralkyl, (a!koxyaryl)alkyl, (sulfonylamino)alkyl (such as (alkyl-S02-amino)alkyl), aminoalkyl, amidoalkyl, (cycloaliphatic)alkyl, or haloalkyl.
[0042] As used herein, an "alkenyl" group refers to an aliphatic carbon group that contains 2-8 (e.g., 2-12, 2-6, or 2-4) carbon atoms and at least one double bond. Like an alkyl group, an alkenyl group can be straight or branched. Examples of an alkenyl group include, but are not limited to allyl, isoprenyl, 2-bulenyl, and 2-hexenyl. An alkenyl group can be optionally substituted with one or more substituents such as halo, phospho, cycloaliphatic [e.g., cycloalkyl or cycloalkenylj, heterocycloaliphatic [e.g., heterocycloalkyl or heterocycloalkenyl], aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl [e.g., (aliphalic)carbonyl, (cycloaliphatic)carbonyl, or (heterocycloaliphatic)carbonylj, nilro, cyano, amido [e.g., (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocyc loalkyl)carbony lam i no, (heterocyc loalky 1 alkyl)carbony I amino, heteroarylcarbonylamino, heteroaralkylcarbonylamino alkylaminocarbonyl, cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl, arylaminocarbonyl, or heteroarylaminocarbonyl ], amino [e.g., aliphaticamino, cycloaliphaticamino, heterocycloaliphaticamino, or aliphaticsulfonylamino], sulfonyl [e.g., alkyl-SOz-, cycloaliphatie-SCh-, or ary]-S02-], sulfinyl, sulf'anyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, carboxy, carbamoyl, cycloaliphalicoxy, hcterocycloaliphaticoxy, aryloxy, heteroaryloxy, aralky!oxy, heteroaralkoxy, alkoxycarbonyl, alkyicarbonyloxy, or hydroxy. Without limitation, some examples of substituted alkenyls include cyanoalkenyl, a!koxyalkenyl, acylalkenyl, hydroxyalkenyl, aralkenyl, (alkoxyaryl)alkenyl, (sulfony]aniino)alkenyl (such as (alkyl-S02-amino)alkenyl), aminoalkenyl, amidoulkenyl, (cyc]oaliphatic)alkenyl, or haloalkenyl.
[0043] As used herein, an "alkynyl" group refers to an aliphatic carbon group that contains 2-8 (e.g., 2-12, 2-6, or 2-4) carbon atoms and has at least one triple bond. An alkynyl group can be straight or branched. Examples of an alkynyl group include, but are not limited to, propargyl and butynyl. An alkynyl group can be optionally substituted with one or more substitucnls such as aroyl, heteroaroyl, alkoxy, cycloalkyloxy, helerocycloalkyloxy, aryloxy, heleroaryloxy, aralkyloxy, nitro, carboxy, cyano, halo, hydroxy, sulfo, mercapto, sulfanyl [e.g., aliphalicsulfanyl or cycloaliphaticsulfanyl], sulfinyl [e.g., aliphaticsulfinyl or cycloaliphau'csulfinyl], sulfonyl [e.g., aliphatic-SCV, aliphaticaniino-SC -, or cycloaliphatic-SO2-], amido [e.g., aminocarbonyl, alkylaniinocarbonyl, alkylcarbonylamino, cycloalkylaniinocarbonyl, heterocycloalkylaminocarbonyl, cycloalkylcarbonylamino, arylaniinocarbonyl, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (cycloalkylalkyl)carbonylamino, heteroaralkylcarbonylamino, heleroarylcarbonylamino or heleroarylaminocarbonylj, urea, thiourea, sulfamoyl, sulfamide, alkoxycarbonyl, alkyicarbonyloxy, cycloaliphatic, heterocycloaliphatic, aryl, heteroaryl, acyl [e.g., (cycloaliphalic)carbonyl or (heterocycloaliphadc)carbonyl], amino [e.g., aliphaticamino], sulfoxy, oxo, carboxy, carbamoyl, (cyc!oaliphatic)oxy, (heterocycloaliphatic)oxy, or (heteroaryl)alkoxy.
[0044] As used herein, an "amido" encompasses both "aminocarbonyl" and "carbonylamino". These terms when used alone or in connection with another group refer lo an amido group such as -N(R )-C(0)- Y or -C(0)-N(Rx)2, when used terminally, and -C(O)-N(RX)- or -N(Rx)-C(0)- when used internally, wherein Rx and RY are defined below.
Examples of amido groups include alkylamido (such as alkylcarbonylamino or alkylaniinocarbonyl), (heterocycloaliphatic)amido, (heteroaralkyl)amido, (heteroaryl)amido, (heterocycloalkyl)alkylamido, arylamido, aralkylamido, (cycloalkyl)alkylamido, or cycloalkylamido.
[0045] As used herein, an "amino" group refers lo -NRxRy wherein each of Rx and RY is independently hydrogen, aliphatic, cycloaliphatic, (cy loaliphatic)aliphatic, aryl, araliphatic, heterocycloaliphatic, (heierocycloalipriatic)aliphatic, heteroaryl, earboxy, sulf'anyl, sulfinyl, sulfonyl, (aliphatic)carbonyl, (cycloaliphatic)carbonyl, ((cycloaliphatic)aliphalic)carbonyl, arykarbonyl, (araliphatic)carbonyl, (heterocycloaliphatic )carbonyl, ((heterocycloaliphalic)aliphatic)carbonyl, (heteroaryl)carbonyl, or (heteroaraliphatic)carbonyl, each of which being defined herein and being optionally substituted. Examples of amino groups include alkylamino, dialkylamino, or arylamino. When the term "amino" is not the terminal group (e.g., alkylcarbonyl amino), it is represented by -NRX-. Rx has the same meaning as defined above.
[0046] As used herein, an "aryl" group used alone or as part of a larger moiety as in "aralkyl", "aralkoxy", or "aryloxyalkyl" refers to monocyclic (e.g., phenyl); bicyclic (e.g., indenyl, naphthalenyl, tetrahydronaphthyl, tetrahydroindenyl); and tricyclic (e.g., fluorenyl tetrahydrofluorenyl, or letrahydroanthracenyl, anthracenyl) ring systems in which the monocyclic ring system is aromatic or at least one of the rings in a bicyclic or tricyclic ring system is aromatic. The bicyclic and tricyclic groups include benzofused 2-3 membered carbocyclic rings. For example, a benzofused group includes phenyl fused with two or more C4-3 carbocyclic moieties. An aryl is optionally substituted with one or more substituents including aliphatic |e.g., alkyl, alkenyl, or alkynyl l; cycloaliphatic; (cycloaliphatic)aliphatic; heterocycloaliphatic; (heterocycloaliphatic)aliphatic; aryl; heteroaryl; alkoxy; (cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy; heteroaryloxy; (araliphalic)oxy; (heteroaraliphatic)oxy; aroyl; heteroaroyl; amino; oxo (on a non-aromadc carbocyclic ring of a benzofused bicyclic or tricyclic aryl); nitio; earboxy; amido; acyl [e.g., (aliphalic)carbonyl; (cycloaliphalic)carbonyl; ((cycloaliphatic)aliphatic)carbonyl; (araliphatic)carbonyl; (heterocycloaliphatic)carbonyl; ((hetcrocycloaliphatic)aliphatic)carbonyl; or (heteroaraliphatic)carbonylj; sulfonyl [e.g., aliphatic-SCV or amino-SCVJ; sulfinyl [e.g., aliphatic-S(O)- or cycloaliphatic-S(O)-]; sulfanyl [e.g., aliphalic-S-J; cyano; halo; hydroxy; mercapto; sulfoxy; urea; thiourea; sulfamoyl; sulf amide; or carbamoyl. Alternatively, an aryl can be unsubstituted.
[0047] Non-limiting examples of substituted aryls include haloaryl [e.g., mono-, di (such as .m-dihaloaryl), and (trihalo)aryl|; (carboxy)aryl [e.g., (alkoxycarbonyl)aryl, ((aralkyl)carbonyloxy)aryl, and (alkoxycarbonyl)arylj; (amido)aryl [e.g., (aminocarbonyl)aryl, (((alkylamino)alkyl)aminocarbonyl)aryl, (alkylcarbonyl)aminoaryl, (arylaminocarbonyl)aryl, and (((heteroaryl)amino)carbonyl)arylj; aminoaryl [e.g., ((alkylsulf'onyl)amino)aryl or ((dialkyl)amino)arylJ; (cyanoalkyl)aryl; (alkoxy)aryl; (sulfamoyl)aryl (e.g., (aminosulfonyl)aryl]; (alk lsulfonyl)aryl; (cyano)aryl; (hydroxyalkyl)aryl; ((alkoxy)alkyl)aryl; (hydroxy)aryl, ({carboxy)alky])aryl; (((dialky])amino)alkyl)aryl; (nitroalkyl)aryl; (((a]kylsulfonyl)amino)alkyl)aryl; ((heterocycloaIiphatic)carbonyl)aryI; ((alkylsu]fonyl)alkyl)aryl; (cyanoalkyl)aryl; (hydr xyalkyl)aryl; (alkylcarbonyl)aryl; alkylaryl; (trihaloalkyl)ary!; p-amino-m-alkoxycarbonylaryl; p-amino-/n-cyanoaryl; j-halo-in-aminoaryl; or (w-(heterocycloaliphatic)-o-(alkyl))aryl.
[0048] As used herein, an "araliphatic" such as an "aralkyl" group refers to an aliphatic group (e.g., a C alkyl group) thai is substituted with an aryl group. "Aliphatic," "alkyl," and "aryl" are defined herein. An example of an araliphatic such as an aralkyl group is benzyl.
[0049] As used herein, an "aralkyl" group refers to an alkyl group (e.g., a CM alkyl group) that is substituted with an aryl group. Both "alkyl" and "aryl" have been defined above. An example of an aralkyl group is benzyl. An aralkyl is optionally substituted with one or more substituents such as aliphatic [e.g., alkyl, alkenyl, or alkynyl, including carboxyalkyl, hydroxyalkyl, or haloalkyl such as tri luoroniethyl], cycloaliphatic [e.g., cycloalkyl or cycloalkcnyl], (cycloalkyl)alkyl, heterocycloalkyl, (heterocycloa!kyl)alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy, hetcrocycloalkyloxy, aryloxy, hcleroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heleroaroyl, nilro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aniido [e.g., aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkylalkyl)carbonylamino, ary!carbonylamino, aralkylcarbonylamino, (heterocyc]oalkyl)carbonylaniino, (hcierocycloalkylalkyl)carbonylaiiiino, heteroarylcarbonylamino, or heteroaralkylcarbonylanrino], cyano, halo, hydroxy, acyl, mercaplo, alkylsulfanyl, sul oxy, urea, thiourea, sulfamoyl, sulfamidc, xo, or carbamoyl.
[0050] As used herein, a "bicyclic ring system" includes 8- 12 (e.g., 9, 10, or 1 1 ) nienibered structures that form two rings, wherein the two rings have at least one atom in common (e.g., 2 atoms in common). Bicyclic ring systems include bicycloaliphatics (e.g., bicycloalkyl or bicycloalkenyl), bicycloheleroaliphatics, bicyclic aryls, and bicyclic heleroaryls.
[0051] As used herein, a "carbocycle" or "cycloaliphatic" group encompasses a "cycloalkyl" group and a "cycloalkcnyl" group, each of which being optionally substituted as set forth below.
[0052] As used herein, a "eycloalkyl" group refers to a saturated carbocyclic mono- or bicyclic (fused or bridged) ring of 3- 10 (e.g., 5-10) carbon atoms. Examples of eycloalkyl groups include cyclopropyl, cyclobulyl, cyclopentyl, cyelohexyl, cycloheptyl, adamantyl, norbornyl, cubyl, octahydro-indenyl, decahydro-naphlhyl, bicyclo[3.2.1]octyl, bicyclo[2.2.21octyl, bicyclo[ 3.3.1 ]nonyl, bicyclo| 3.3.2.1deeyl, bicyclo[2.2.2|octyl, adamantyl, or (( m i nucarbony l)cy c loalk I )cyc 1 oalky 1.
[0053] Λ "cycloalkenyl" group, as used herein, refers to a non-aromatic carbocyclic ring of 3- 10 (e.g., 4-8) carbon atoms having one or more double bonds. Examples of cycloalkenyl groups include cyclopentenyl, 1 ,4-cyclohexa-di-enyl, cycloheptenyl, cyclooctenyl, he ahydro-indenyl, octahydro-naphthyl, cyclohexenyl, cyclopentenyl, bicyclo[2.2.2]octenyl, or bicyclo|3.3.1 Jnonenyl.
[0054] A eycloalkyl or cycloalkenyl group can be optionally substituted with one or more substitucnts such as phosphor, aliphatic [e.g., alky], alkcnyl, or alkynylj, cycloaliphatic, (cycloaliphatic) aliphatic, helcrocycloaliphatic, (helerocycloaliphatic) aliphatic, aryl, heteroaryl, alkoxy, (cycloaiiphalic)oxy, (heterocycloaliphatic)oxy, aryloxy, heteroaryloxy, (araliphatic)oxy, (hcteroaraliphattc)oxy, aroyl, hctcroaroyl, amino, amido [e.g., (aliphatic)carbonylamino, (cycloaliphalic)carbonylamino, ((cycloaliphalic)aliphatic)carbonylamino, (aryl)carbonylamino, (araliphatic)carbonylamino, (hcterocycloaliphatic)carbonylamino, ((hctcrocycloaliphalic)aliphaiic)carbonylaniino, (heteroaryl)carbonylamino, or (heteroaraliphalic)carbonylauiinoj, nitro, carboxy |e.g., HOOC-, alkoxycarbonyl, or alkylcarbonyloxy], acyl [e.g., (cycloaliphatic)carbonyl, ((cycloaliphatic) aliphatic)carbonyl, (araliphatic)carbonyl, (heterocycloaliphatic)carbonyl, ((heterocycloaliphatic)aliphatic)carbonyl, or (heteroaraliphatic)carbonyl], cyano, halo, hydroxy, mercapto, sulfonyl le.g., alkyl-SC>2- and aryl-S02-], sulfinyl [e.g., alkyl-S(0)- |, sulfanyl [e.g., alkyl-S-], sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, or carbamoyl.
[0055] As used herein, the term "heterocycle" or "helerocycloaliphatic" encompasses a heterocycloalkyl group and a heterocycloalkenyl group, each of which being optionally substituted as set forth below.
[0056] As used herein, a "heterocycloalkyl" group refers to a 3-10 membered mono- or bicylic (fused or bridged) (e.g., 5- to 10-membered mono- or bicyclic) saturated ring structure, in which one or more of the ring atoms is a heteroalom (e.g., N, O, S, or combinations thereof). Examples of a heterocycloalkyl group include piperidyl, piperazyl, tetrahydropyranyl, tetrahydrofuryl, l ,4-dioxo3anyl, 1 ,4-dithianyl, 1 ,3-dioxolanyl, oxazolidyl, isoxazolidyl, tra^holinyl, thiomoφhol l) octahydrobenzofuryl, octahydrochromenyl, octahydrothiochroinenyl, oclahydroindolyl, octahydropyrindinyl, decahydroquinoliny], octahydrobenzo[i>]thiopheneyl, 2-oxa-bicyclo[2.2.2]octyl, 1 -aza-bicyclo[2.2.2]octyl, 3-aza-bicyclo[3.2.1]octyl, and 2,6-dioxa-tricyclo[3.3.1.03'7]nonyl. A monocyclic heterocycloalkyl group can be fused with a phenyl moiety to form structures, such as tetrahydroisoquinoline, which would be categorized as heteroaryls.
[0057] A "hetcrocycloalkcnyl" group, as used herein, refers to a mono- or bicylic (e.g., 5-to 10-nienibered mono- or bicyclic) non-aromatic ring structure having one or more double bonds, and wherein one or more of the ring atoms is a heteroatom (e.g., N, O, or S).
Monocyclic and bicyclic heterocycloaliphatics are numbered according to standard chemical nomenclature.
[0058] A heterocycloalkyl or heterocycloalkenyl group can be optionally substituted with one or more substituents such as phosphor, aliphatic [e.g., alkyl, alkenyl, or alkynyl], cycloaliphatic, (cycloaliphalic)aliphatic, heterocycloaliphalic, (heterocycloalipha-ic)aliphatie, aryl, heteroary], alkoxy, (cycloaliphatic)oxy, (heierocycloaliphatic)oxy, aryloxy, hctcroaryloxy, (araliphatic)oxy, (hctcroaraliphatic)oxy, aroyl, hetcroaroyl, amino, amido [e.g., (aliphalic)carbonylamino, (cycloaliphatic)carbonylamino, ((cycloaliphatic) aliphatic)carbonylamino, (aryl)carbonylamino, (araliphatic)carbonylamino, (hctcrocycloaliphatic)carbonylamino, ((hcterocyclo aliphatic) aliphatic)carbonylamino, (heteroaryi)carbonylamino, or (heleroaraliphatic)carbonylaminoj, nitro, carboxy [e.g., HOOC-, alkoxycarbonyl, or alkylcarbonyloxy], acyl [e.g., (cycloaliphatic)carbonyl, ((cycloaliphatic) aliphatic)carbonyl, (araliphatic)carbonyl, (heterocycloaliphatic)carbonyl, ((heterocycloaliphatic)aliphatic)carbonyl, or (heteroaraliphatic)carbonylj, nitro, cyano, halo, hydroxy, mercapto, sulfonyl [e.g.. alkylsulfonyl or arylsulfonyl], sulfinyl [e.g., alkylsulfinyl], sulfanyl [e.g., alkylsulfanyl], sul foxy, urea, thiourea, sulfamoyl, sulfamide, oxo, or carbamoyl.
[0059] A "heteroaryl" group, as used herein, refers to a monocyclic, bicyclic, or tricyclic ring system having 4 to 15 ring atoms wherein one or more of the ring atoms is a heteroatom (e.g., N, O, S, or combinations thereof) and in which the monocyclic ring system is aromatic or at least one of the rings in the bicyclic or tricyclic ring systems is aromatic. A heteroaryl group includes a benzofused ring system having 2 to 3 rings. For example, a benzofused group includes benzo fused with one or two 4 to 8 membered heterocycloaliphatic moieties (e.g., indolizyl, indolyl, isoindolyl, 311-indolyl, indolinyl, benzo[&]furyl, benzo[6]thiophenyl, quinolinyl, or isoquinolinyl). Some examples of heteroaryl are azetidinyl, pyridyi, 1 H-i ndazolyl, furyl, pyrrolyl, ihicnyl, thiazolyl, oxazoiyl, imidazolyl, letrazolyl, benzo fur l, isoquinolinyl, benzlhiazolyl, xanlhene, thioxanihene, phenothiazine, dihydroindole, benzol 1 ,3 jdioxole, benzo[b]furyl, benzo[b]thiophenyl, indazolyl, benzimidazolyl, benzlhiazolyl, puryl, cinnolyl, quinolyl, quinazolyl,cinnolyl, phlhalazyl, quinazolyl, quinoxalyl, isoquinolyl, 4H-quinolizyl, benzo- l,2,5-lhiadiazolyl, or 1 ,8-naphthyridyl.
[0060] Without limitation, monocyclic helcroaryls include furyl, ihiophcnyl, 211-pyrrolyl, pyrrolyl, oxazoiyl, thazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1,3,4-lhiadiazolyl, 2H-pyranyl, 4-H-pranyl, pyridyi, pyridazyl, pyrimidyl, pyrazolyl, pyrazyl, or 1,3,5-triazyl. Monocyclic helcroaryls are numbered according to standard chemical nomenclature.
[0061] Without limitation, bicyclic heteroaryls include indolizyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo[/?]furyl, benzo[&]thiophenyl, quinolinyl, isoquinolinyl, indolizinyl, isoindolyl, indolyl, bcnzo[b] furyl, bcxo[fr]thiophcnyl, indazolyl, bcnzimidazyl, benzlhiazolyl, purinyl, 4H-quinolizyl, quinolyl, isoquinolyl, cinnolyl, phlhalazyl, quinazolyl, quinoxalyl, 1 ,8-naphthyridyl, or pteridyl. Bicyclic heteroaryls are numbered according to standard chemical nomenclature.
[0062] A heteroaryl is optionally substituted with one or more substituents such as aliphatic [e.g., alkyl, alkenyl, or alkynylj; cycloaliphatic; (cycloaliphatic)aliphatic; heterocycloaliphatic; (heterocycloaliphatic)aliphatic; aryl; heteroaryl; alkoxy; (cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy; heteroaryloxy; (araliphalic)oxy; (heteroaraliphatic)oxy; aroyl; heteroaroyl; amino; oxo (on a non-aromatic carbocyclic or heterocyclic ring of a bicyclic or tricyclic heteroaryl); carboxy; amido; acyl [ e.g., aliphaticcarbonyl; (cycloaliphatic)carbonyl; ((cycloaliphatic)aliphatic)carbonyl; (arali ph atic)carbony 1 ; (he terocycloaliphati c)carbony 1 ; ((heterocycloaliphatic)aliphatic)carbonyl; or (heteroaraliphatic)carbonyl]; sulfonyl [e.g., aliphaticsulfonyl or aminosulfonylj; sulfinyl [e.g., aliphaticsulfinylj; sulfanyl [e.g., aliphaticsulfanyl]; nitro; cyano; halo; hydroxy; mercapto; sulfoxy; urea; thiourea; sulfamoyl; sulfamidc; or carbamoyl. Alternatively, a heteroaryl can be unsubstitutcd.
[0063] Non-limiting examples of substituted heteroaryls include (halo)heteroaryl [e.g., mono- and di-(halo)heteroaryl]; (carboxy)heleroaryi [e.g., (alkoxycarbonyl)heleroaryl]; cyanoheteroaryl; aminoheleroaryl |e.g., ((alkylsulfonyl)amino)heteroaryl and ((dialkyl)amino)heteroaryl |; (amido)heteroaryl [e.g., aminocarbonylheteroaryl, ((alkylcarbonyl)amino)heteroaryl, ((((alkyl)amino)alkyl)aminocarbonyl)heleroaryl, (((heteroaryl)amino)carbonyl)heteroaryl, ((helerocycloaliphatic)carbonyl)heteroaryl, and ((alkylcarbonyDamino)heleroaryl I; (cyanoalk l)hetcroaryl; (alkoxy)heteroaryl; (sulfamoyl)heteroaryl [e.g., (aminosulfonyl)heteroaryl]; (sulfonyl)heteroaryl [e.g., (alkylsulfonyl)heleroaryl]; (hydroxyalkyl)heteroaryl; (alkoxyalkyl)heteroaryl; (hydroxy)heteroaryl; ((carboxy)alkyl)hetcroaryl; (((dialkyl)aniino)alkyl]heteroaryl; (heterocycloaliphatic)heteroaryl; (cycloaliphatic)heteroaryl; (nitroalkyl)heteroaryl; (((alkylsulfonyl)amino)alkyl)heteroaryl; ((alky]sulfonyl)alkyl)heteroaryl; (cyanoalkyl) heteroaryl; (acy!)heteroaryl [e.g., (alkylcarbonyl)heteroarylj; (alkyl)heteroaryl, and (haloalkyl)hctcroaryl [e.g., trihaloalkylhctcroarylj.
[0064] A "heteroaraliphatic" (such as a heteroaralkyl group) as used herein, refers to an aliphatic group (e.g., a C alkyl group) that is substituted with a heteroaryl group.
"Aliphatic," "alkyl," and "heteroaryl" have been defined above.
[0065] A "hctcroaralkyr group, as used herein, refers to an alky! group (e.g., a C alkyl group) that is substituted with a heteroaryl group. Both "alkyl" and "heteroaryl" have been defined above. A heteroaralkyl is optionally substituted with one or more subslituents such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl such as trifluoromethyl), alkenyl, alkynyl, cycloalkyl, (cyclo lky l)a Iky 1, heterocycloalkyl, (heterocycloalkyl)alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy, helerocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, hcteroaralkyloxy, aroyl, hctcroaroyl, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylaniino, (cycloa Iky 1 alk l)c arbon y lam i no, ar lcarbony lam i no , aral k lcarbony 1 am i no , (heterocycloalkyl)carbonylamino, (heteroeycloalkylalkyl)carbony]amino, heteroarylcarbonylaniino, heteroaralkylcarbonylamino, cyano, halo, hydroxy, acyl, mercapto, alkylsulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, or carbamoyl.
[0066] As used herein, "cyclic moiety" and "cyclic group" refer to mono-, bt-, and tricyclic ring systems including cycloaliphatic, heterocycloaliphalic, ary], or heteroaryl, each of which has been previously defined.
[0067] As used herein, a "bridged bicyciic ring system" refers to a bicyciic heterocyclicaliphatic ring system or bicyciic cycloaliphatic ring system in which the rings are bridged. Examples of bridged bicyciic ring systems include, but are not limited to, adamantanyl, norbornanyl, bicyclo[3.2.1 |octyl, bicyclo[2.2.2]octyl, bicyclo[3.3.1 jnonyl, bicyclo[3.2.3]nony], 2-oxabicyclo|2.2.2]octyl, l -azabicyclo[2.2.2]octyl, 3-azabicyclo|3.2. Ijoctyl, and 2,6-dioxa-tricyclo[3.3. 1.03,7]nonyl. A bridged bicyciic ring system can be optionally substituted with one or more subsliluenls such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl such as trifluoromelhyl), alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy, helcrocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, hctcroaroyl, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino, heleroaralkylcarbonylamino, cyano, halo, hydroxy, acyl, mcrcapto, alk lsulf any 1, sulfoxy, urea, thiourea, sullamoyl, sulfamide, oxo, or carbamoyl.
[0068] As used herein, an "acyl" group refers lo a formyl group or Rx-C(0)- (such as alkyl-C(O)-, also referred to as "alkylcarbonyl") where R and "alkyl" have been defined previously. Acetyl and pivaloyl are examples of acyl groups.
[0069] As used herein, an "aroyl" or "hctcroaroyl" refers to an aryl-C(O)- or a heleroaryl-C(O)-. The aryl and heteroaryl portion of the aroyl or heteroaroyl is optionally substituted as previously defined.
[0070] As used herein, an "alkoxy" group refers to an alkyl-O- group where "alkyl" has been defined previously.
[0071] As used herein, a "carbamoyl" group refers to a group having the structure -0-CO-NR RY or -NR -CO-0-Rz, wherein Rx and RY have been defined above and Rz can be aliphatic, aryl, araliphatic, heterocycloaliphatic, heteroaryl, or heleroaraliphatic.
[0072] As used herein, a "carboxy" group refers to -COOH, -COORx, -OC(0)H, -OC(0)Rx, when used as a terminal group; or -OC(O)- or -C(0)0- when used as an internal group.
[0073] As used herein, a "haloaliphalic" group refers to an aliphatic group substituted with 1 -3 halogen. For instance, the term haloalkyl includes the group -CF3.
[0074] As used herein, a "mercapto" group refers to -S1 I.
[0075] As used herein, a "sulfo" group refers to -SO3H or -S03RX when used terminally or -S(0)3- when used internally.
[0076] As used herein, a "sulfamide" group refers to the structure -NRX-S(0)2-NRYRZ when used terminally and -NR -S(0)2-NRY- when used internally, wherein Rx, RY, and Rz have been defined above.
[0077] As used herein, a "sulfonamide" group refers to the structure -S(0)2-NRxRY or -NR -S(0)2-Rz when used teniiinally; or -S(0)2-NRx- or -NRX -S(0)2- when used internally, wherein R , RY, and Rz are defined above.
[0078] As used herein a "sulfanyl" group refers to -S-Rx when used terminally and -S-when used internally, wherein R has been defined above. Examples of sulfanyls include aliphatic-S-, cycloaliphatic-S-, aryl-S-, or the like.
[0079] As used herein a "sulfinyl" group refers to -S(0)-Rx when used terminally and -S(O)- when used internally, wherein Rx has been defined above. Exemplary sulfinyl groups include aliphatic-S(O)-, aryl-S(O)-, (cycloaliphatic(aliphatic)>S(0)-, cycloalkyl-S(O)-, heterocycloaliphatic-S(O)-, heleroaryl-S(O)-, or the like.
[0080] As used herein, a "sulfonyl" group refers to-S(0)2-Rx when used tenninally and -S(0)2- when used internally, wherein R has been defined above. Exemplary sulfonyl groups include aliphatic-S(0)2-, aryl-S(0)2-, (cycloaliphatic(aliphatic))-S(0)2-, cycloaliphatic-S(0)2-, heterocycloaliphatic-S(0)2-, heteroaryl-S(0)2-, (cycloaliphatic(ainido(aliphatic)))-S(0)2-or the like.
[0081] As used herein, a "sulfoxy" group refers to -0-SO-Rx or -SO-0-Rx, when used teniiinally and -O-S(O)- or -S(0)-0- when used internally, where Rx has been defined above.
[0082] As used herein, a "halogen" or "halo" group refers to fluorine, chlorine, bromine or iodine.
[0083] As used herein, an "alkoxycarbonyl," which is encompassed by the term carboxy, used alone or in connection with another group refers to a group such as alkyl-O-C(O)-.
[0084] As used herein, an "alkoxyalkyl" refers to an alkyl group such as alkyl-O-alkyl-, wherein alkyl has been defined above.
[0085] As used herein, a "carbonyl" refer to -C(O)-.
[0086] As used herein, an "oxo" refers to =0.
[0087] As used herein, the term "phospho" refers to phosphinates and phosphonates. Examples of phosphinates and phosphonates include -P(0)(Rp)2, wherein R is aliphatic, alkoxy, aryloxy, heteroaryloxy, (cycloaliphatic)oxy, (heterocycloaliphatic)oxy aryl, heteroaryl, cycloaliphatic or amino.
[0088] As used herein, an "aminoalkyl" refers to the structure (Rx)2N-alkyI-.
[0089] As used herein, a "cyanoalkyl" refers to the structure (NC)-alkyl-.
[0090] As used herein, a "urea" group refers to the structure -NRx-CO-NRYRz and a "thiourea" group refers to the structure -NRX-CS-NRYRZ when used terminally and -NRX-CO-NRY- or -NR -CS-NRY- when used internally, wherein Rx, RY, and Rz have been defined above,
[0091] As used herein, a "guanidine" group refers to the structure -N=C(N(RXRY))N(R RY) or -NRX-C(=NRX)NRXRY wherein Rx and RY have been defined above.
[0092] As used herein, the term "amidino" group refers to the structure -0(NRx)N(RxRY) wherein R and RY have been defined above.
[0093] In general, the term "vicinal" refers to the placement of substiluents on a group that includes two or more carbon atoms, wherein the substitucnts are attached to adjacent carbon atoms.
[0094] In general, the term "geminal" refers to the placement of substitucnts on a group that includes two or more carbon atoms, wherein the substituents are attached to the same carbon atom.
[0095] 'J ie terms "terminally" and "internally" refer to the location of a group within a substituent. A group is terminal when the group is present at the end of the substituent not further bonded to the rest of the chemical structure. CarboxyalkyI, i.e., R O(0)C-alkyI is an example of a carboxy group used terminally. A group is internal when the group is present in the middle of a substituent of the chemical structure. Alkylcarboxy (e.g., alkyl-C(0)0- or alkyl-OC(O)-) and alkylcarboxyaryl (e.g., alkyl-C(0)0-aryl- or alkyl-O(CO)-aryl-) are examples of carboxy groups used internally.
[0096] As used herein, an "aliphatic chain" refers to a branched or straight aliphatic group (e.g., alky] groups, alkenyl groups, or alkynyl groups). A straight aliphatic chain has the structure - [CH2]v-, where v is 1-12. A branched aliphatic chain is a straight aliphatic chain that is substituted with one or more aliphatic groups. A branched aliphatic chain has the structure -I QQIv- where each Q is independently a hydrogen or an aliphatic group; however, Q shall be an aliphatic group in at least one instance. The term aliphatic chain includes alkyl chains, alkenyl chains, and alkynyl chains, where alkyl, alkenyl, and alkynyl are defined above.
[0097] rITie phrase "optionally substituted" is used interchangeably with the phrase "substituted or unsubstitutcd." As described herein, compounds of the invention can opu'onally be substituted with one or more substituents, such us are illustrated generally above, or as exemplified by particular classes, subclasses, and species of the invention. As described herein, the variables Ri, R2, and R3, and other variables contained in formulae described herein encompass specific groups, such as alkyl and aryl. Unless otherwise noted, each of the specific groups for the variables R], R2, and R3, and other variables contained therein can be optionally substituted with one or more substituents described herein. Each substituent of a specific group is further optionally substituted with one to three of halo, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl, cycloaliphatic, heterocycloaliphatic, hcteroary], haloalkyl, and alkyl. For instance, an alkyl group can be substituted with alkylsulfanyl and the alkylsulfanyl can be optionally substituted with one to three of halo, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl, haloalkyl, and alkyl. As an additional example, the cycloalkyl portion of a (cycloalkyl)carbonylamino can be optionally substituted with one to three of halo, cyano, alkoxy, hydroxy, nitro, haloalkyl, and alkyl. When two alkoxy groups are bound to the same atom or adjacent atoms, the two alkxoy groups can form a ring together with the atom(s) to which they are bound.
[0098] In general, the term "substituted," whether preceded by the term "optionally" or not, refers to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. Specific substituents are described above in the definitions and below in the description of compounds and examples thereof. Unless otherwise indicated, an optionally substituted group can have a substituent at each substilutable position of the group, and when more than one position in any given structure can be substituted with more than one substituent selected from a specified group, the substituent can be either the same or different at every position. A ring substituent, such as a heterocycloalkyl, can be bound to another ring, such as a cycloalkyl, to form a spiro-bicyclic ring system, e.g., both rings share one common atom. As one of ordinary skill in the art will recognize, combinations of substituents envisioned by this invention are those combinations that result in the formation of stable or chemically feasible compounds.
[0099] The phrase "stable or chemically feasible," as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and preferably their recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, a stable compound or chemically feasible compound is one that is not substantially altered when kept at a temperature of 40 °C or less, in the absence of moisture or other chemically reactive conditions, for at least a week.
[00100] As used herein, an "effective amount" is defined as the amount required to confer a therapeutic effect on the treated patient, and is typically determined based on age, surface area, weight, and condition of the patient. rI¾e interrelationship of dosages for animals and humans (based on milligrams per meter squared of body surface) is described by Freireich et al., Cancer Chemother. Rep., 50: 219 ( 1966). Body surface area may be approximately determined from height and weight of the patient. See, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley, New York, 537 (1970). As used herein, "patient" refers to a mammal, including a human.
[00101] Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and S configurations for each as mmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. 'ITierefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a , 3C- or 14C-enriched carbon are within the scope of this invention. Such compounds are useful, for example, as analytical tools or probes in biological assays, or as therapeutic agents.
[00102] Compounds of the present invention are useful modulators of ABC transporters and are useful in the treatment of ABC transporter mediated diseases.
[00103] 11. COMPOUNDS
[00104] A. Generic Compounds
[00105] The present invention relates to compounds of formula I useful as modulators of ABC transporter activity: I or a pharm ceuticall acceptable salt thereof.
[00106] R) is -ZAR-!, wherein each ZA is independently a bond or an optionally substituted branched or straight Ci.6 aliphatic chain wherein up to two carbon units of ZA are optionally and independently replaced by -CO-, -CS-, -CONRA-, -CONRANRA-, -COa-, -OCO-, -NRAC02-, -0-, -NRACONRA-, -OCONRA-, -NRANRA-, -NRACO-, -S-, -SO-, -S02-, -NRA-, -S02NRA-, -NRAS02-, or -NRAS02NRA-. Each 4 is independently RA, halo, -OH, -NH2, -N02, -CN, or -OCF3. Each RA is independently hydrogen, an optionally substituted aliphatic, an optionally substituted cycloaliphatic, an optionally substituted hetcrocycloaliphatic, an optionally substituted aryl, or an optionally substituted heteroaryl.
[00107] R2 is -ZnR5, wherein each ZB is independently a bond or an optionally substituted branched or straight C 1.6 aliphatic chain wherein up to two carbon units of Z are optionally and independently replaced by -CO-, -CS-, -CONRB-, -CONRBNRB-, -C02-, -OCO-, -NRHC02-, -0-, -NRBCONRB-, -OCONR8-, -NRBNRB-, -NRBCO-, -S-, -SO-, -S02-, -NRB-, -S02NRB-, -NRBS02-, or -NRBS02NRB-. Each R5 is independently RB, halo, -OH, -NH2, -N02, -CN, -CF3, or -OCE3. Each RB is independently hydrogen, an optionally substituted aliphatic, an optionally substituted cycloaliphatic, an optionally substituted helerocycloaJiphatic, an optionally substituted aryl, or an optionally substituted heteroaryl. Alternati ely, any two adjacent R2 groups together with the atoms to which they are attached fomi an optionally substituted carbocycle or an optionally substituted heterocycle.
[00108] Ring A is an optionally substituted 3-7 mcinbcrcd monocyclic ring having 0-3 heteroatonis selected from N, O, and S.
[00109] Ring B is a group having formula la: la or a phamiaceutically acceptable salt thereof, wherein p is 0-3 and each R3 and R^ is independently -ZCR6, where each Zc is independently a bond or an optionally substituted branched or straight Cj.6 aliphatic chain wherein up to two carbon units of Zc are optionally and independently replaced by -CO-, -CS-, -CONRc-, -CONRcNRc-, -C02-, -OCO-, -NRcC02-, -0-, -NRcCONRc-, -OCONRc-, -NRCNRC-, -NRcCO-, -S-, -SO-, -SO2-, -NRC-, -S02NRc-, -NRcS02-, or -NR S02NR -. Each R6 is independently R , halo, -OH, -NH2, -N02, -CN, or -OCF3. Each Rc is independently hydrogen, an optionally substituted aliphatic, an optionally substituted cycloaliphatic, an optionally substituted heterocycloaliphatic, an optionally substituted aryl, or an optionally substituted hetcroaryl. Alternatively, any two adjacent R3 groups together with the atoms to which they are attached form an optionally substituted carbocycle or an optionally substituted heterocycle, Furthermore, 3 and an adjacent R3 group, together with the atoms to which they arc attached, form an optionally substituted heterocycle.
[00110] n is 1 -3.
[00111] However, in several embodiments, when ring A is unsubstituted cyclopentyl, n is 1 , R2 is 4-chloro, and i is hydrogen, then ring B is not 2-(tcrtbutyl)indol-5-yl, or (2,6-dichloropheny](carbonyl))-3-melhyl-lH-indol-5-yl; and when ring A is unsubstituted cyclopentyl, n is 0, and Ri is hydrogen, then ring B is not
[00112] B. Specific Compounds
[00113] 1. Rj Group
[00114] | is -ZAR4, wherein each ZA is independently a bond or an optionally substituted branched or straight Ci^ aliphatic chain wherein up to two carbon units of ZA are optionally and independently replaced by -CO-, -CS-, -CONRA-, -CONRANRA-, -CO2-, -OCO-, -NRAC02-, -Ο-, -NRACONRA-, -OCONRA-, -NRANRa-, -NRACO-, -S-, -SO-, -SO2-, -NRA-, -S02NRA-, -NRAS02-, or -NRAS02NRA-. Each R4 is independently RA, halo, -OH, -NH2l -N02, -CN, or -OCF3. Each RA is independently hydrogen, an optionally substituted aliphatic, an optionally substituted cycloaliphatic, an optionally substituted heterocycloaliphatic, an optionally substituted aryl, or an optionally substituted heteroaryi.
[00115] In several embodiments, R] is -ZAR_i, wherein each ZA is independently a bond or an optionally substituted branched or straight Ci^ aliphatic chain and each R4 is hydrogen.
[00116] In other embodiments, Ri is -ZAR4, wherein each ZA is a bond and each R4 is hydrogen.
[00117] 2. R2 Grou
[00118] Each R2 is independently -Z Rs, wherein each Zu is independently a bond or an optionally substituted branched or straight C aliphatic chain wherein up to two carbon units of ZB are optionally and independently replaced by -CO-, -CS-, -C0NR8-, -CONRBNRB-, -CO , -OCO-, -NRuC02-, -0-, -NRBCONRB-, -OCONR1'-, -NRBNRU-, -NRBCO-, -S-, -SO-, -S02-, -NRB-, -S02NRB-, -NRBS02-, or -NRBS02NRB-. Each R5 is independently RB, halo, -OH, -NH2, -N 2, -CN, -CF3, or -OCF3. Each Rn is independently hydrogen, an optionally substituted aliphatic, an optionally substituted cycloaliphatic, an optionally substituted heterocycloaliphatic, an optionally substituted aryl, or an optionally substituted heteroaryi. Alternatively, any two adjacent R2 groups together with the atoms to which they are attached fonn an optionally substituted carbocycle or an optionally substituted heterocycle.
[00119] In several embodiments, R2 is an optionally substituted aliphatic. For example, R2 is an optionally substituted branched or straight Cj.6 aliphatic chain. In other examples, R2 is an optionally substituted branched or straight Ci-s alkyl chain, an optionally substituted branched or straight C2-0 alkenyl chain, or an optionally substituted branched or straight C2-6 alkynyl chain. In alternative embodiments, R2 is a branched or straight Ci^ aliphatic chain that is optionally substituted with 1 -3 of halo, hydroxy, cyano, cycloaliphatic, heterocycloaliphatic, aryl, heteroaryi, or combinations thereof. For example, R2 is a branched or straight alkyl that is optionally substituted with 1 -3 of halo, hydroxy, cyano, cycloaliphatic, heterocycloaliphatic, aryl, heteroaryi, or combinations thereof. In still other examples, R2 is a methyl, ethyl, propyl, butyl, isopropyl, or tert-butyl, each of which is optionally substituted with 1-3 of halo, hydroxy, cyano, aryl, heteroaryi, cycloaliphatic, or heterocycloaliphatic. In still other examples, R2 is a methyl, ethyl, propyl, butyl, isopropyl, or tert-butyl, each of which is unsubstituted.
[00120] In several other embodiments, R2 is an optionally substituted branched or straight C]-5 alkoxy. For example, R2 is a Cj.5 alkoxy that is optionally substituted with 1 -3 of hydroxy, aryl, heteroaryl, cycloaliphatic, heterocycloaliphatic, or combinations thereof. In other examples, R2 is a methoxy, ethoxy, propoxy, butoxy, or pentoxy, each of which is optionally substituted with 1-3 of hydroxy, aryl, heteroaryl, cycloaliphatic, heterocycloaliphatic, or combinations thereof.
[00121] In other embodiments, R2 is hydroxy, halo, or cyano.
[00122] In several embodiments, R2 is -ZBRs, and ZB is independently a bond or an optionally substituted branched or straight C M aliphatic chain wherein up to two carbon units of ZB are optionally and independently replaced by -C(O)-, -0-, -S-, -S(0)2-, or -NH-, and R5 is RB, halo, -OH, -NH2, -NO2, -CN, -CF3, or -OCF.,, and RB is hydrogen or aryl.
[00123] In several embodiments, two adjacent R2 groups form an optionally substituted carbocycle or an optionally substituted heterocycle. For example, two adjacent R2 groups form an optionally substituted carbocycle or an optionally substituted heterocycle, either of which is fused to the phenyl of I'onnula 1, wherein the carbocycle or heterocycle has formula lb: lb
[00124] Each of Zi, Z2, Z3( Z4, and Z5 is independently a bond, -CR7RV, -NR7-, or -0-; each R7 is independently -ZDRs, wherein each ZD is independently an optionally substituted branched or straight C1. aliphatic chain wherein up to two carbon units of ZD are optionally and independently replaced by -CO-, -CS-, -CONR0-, -CO2-, -OCO-, -NRDC02-, -0-, -NRDCONR -, -OCONR0-, -NR NRD-, -NR CO-, -S-, -SO-, -S02-, -NRD-, -S02NRD-, -NRDS02-, or -NRDS02NRD-. Each Re is independently RD, halo, -OH, -NH2, -N02, -CN, -CF3, or -OCF3. Each RD is independently hydrogen, an optionally substituted cycloaliphatic, an optionally substituted heterocycloaliphatic, an optionally substituted aryl, or an optionally substituted heteroaryl. Each R'7 is independently hydrogen, optionally substituted Ci-e aliphatic, hydroxy, halo, cyano, nitro, or combinations thereof. Alternatively, any two adjacent R7 groups together with the atoms to which they are attached form an optionally substituted 3-7 membered carbocyclic ring, such as an optionally substituted cyclobutyl ring, or any two R? and R'7 groups together with the atom or atoms to which they are attached fonn an optionally substituted 3-7 membered carbocyclic ring or a heterocarbocyclic ring.
[00125] In several other examples, two adjacent R2 groups form an optionally substituted carbocycle. For example, two adjacent 2 groups fonn an optionally substituted 5-7 membered carbocycle that is optionally substituted with 1-3 of halo, hydroxy, cyano, oxo, cyano, alkoxy, alkyl. or combinations thereof. In another example, two adjacent R2 groups form a 5-6 membered carbocycle that is optionally substituted with 1-3 of halo, hydroxy, cyano, oxo, cyano, alkoxy, alkyl, or combinations thereof. In still another example, two adjacent R2 groups fonn an unsubstituted 5-7 membered carbocycle.
[00126] In alternative examples, two adjacent 2 groups form an optionally substituted heterocycle. For instance, two adjacent R2 groups form an optionally substituted 5-7 membered heterocycle having 1 -3 heleroatoms independently selected from N, 0, and S. In several examples, two adjacent R2 groups form an optionally substituted 5-6 membered heterocycle having 1 -2 oxygen atoms. In other examples, two adjacent R2 groups form an unsubstituted 5-7 membered heterocycle having 1 -2 oxygen atoms. In other embodiments, two adjacent 2 groups fonn a heterocyclic ring selected from: XA7 XA8 XA9 XA10 XA11 XA12 XA19 XA20 XA21
[00127] In alternative examples, two adjacent R2 groups form an optionally substituted carbocycle or an optionally substituted heterocyclc, and a third R2 group is attached to any chemically feasible position on the phenyl of formula I. For instance, an optionally substituted carbocycle or an optionally substituted heterocycle, both of which is formed by two adjacent R2 groups ; a third R2 group; and the phenyl of formula I form a group having formula Ic: Ic
[00128] Zi , Z2, Z3, Z,, and Z5 has been defined above in fonnula lb, and R2 has been defined above in formula I .
[00129] In several embodiments, each R2 group is independently selected from hydrogen, halo, -OCH3, -OH, -CH2OI-I, -CI I3, and -OCF3, and/or two adjacent R2 groups together with the atoms to which they are attached form XA1 XA2 XA3 XA4 XA5 XA6 XA19 XA20 XA21
[00130] In other embodiments, R2 is at least one selected from hydrogen, halo, mcthoxy, phenylniethoxy, hydroxy, hydroxymethyl, trifluoromethoxy, and methyl.
[00131] In some embodiments, two adjacent R2 groups, together with the atoms to which they are attached, form XA1 XA2
[00132] 3. Ring A
[00133] Ring A is an optionally substituted 3-7 membered monocyclic ring having 0-3 heteroatoms selected from N, O, and S.
[00134] In several embodiments, ring A is an optionally substituted 3-7 membered monocyclic cycloaliphatic. For example, ring A is a cyclopropyl, cyclobutyl, cyclopenlyl, cyclohexyl, or cycloheplyl, each of which is optionally substituted with 1 -3 of halo, hydroxy, Ci_5 aliphatic, or combinations thereof.
[00135] In other embodiments, ring A is an optionally substituted 3-7 membered monocyclic heterocycloahphatic . For example, ring A is an optionally substituted 3-7 membered monocyclic heterocycloahphatic having 1-2 heteroatoms independently selected from N, O, and S. In other examples, ring Λ is tetrahydroliiran-yl, telrahydro-2H-pyran-yl, pyrrolidone-yl, or piperidine-yl, each of which is optionally substituted.
[00136] Ln still other examples, ring Λ is selected from
[00137] Each R« is independently -Z^Rg, wherein each ZL is independently a bond or an optionally substituted branched or straight Cj-s aliphatic chain wherein up to two carbon units of ZE are optionally and independently replaced by -CO-, -CS-, -CONRE-, -C02-, -OCO-, -NR¾02-, -0-, -NR¾()NRK-, -OCONRE-, -NRBNRE-, -NRECO-. -S-, -SO-, -SO2-, -NRB-, -S02NRE-, -NR¾02-, or -NRFlS02NRB-, each R9 is independently RB, -OH, -NH2> -N02, -CN, -CF3, oxo, or -OCF3. Each RFC is independently hydrogen, an optionally substituted cycloaliphalic, an optionally substituted heterocycioaliphatic, an optionally substituted aryl, or an optionally substituted heteroaryl.
[00138] q is 0-5.
[00139] In other embodiments, ring Λ is one selected from
[00140] In several embodiments, ring A is
[00141] 4. Ring B
[00142] Ring B is a group having for la or a pharmaceutically acceptable salt thereof, wherein p is 0-3.
[00143] Each R and R'3 is independently - CR6, where each Zc is independently a bond or an optionally substituted branched or straight Ci-& aliphatic chain wherein up to two carbon units of Zc are optionally and independently replaced by -CO-, -CS-, -CONR -, -CONRcNRc-, -CO2-, -OCX)-, -NRcC02-, -0-, -NRcCONRc-, -OCONRc-, -NRCNRC-, -NRcCO-, -S-, -SO-, -S02-, -NRC-, -S02NRc-, -NRcS02-, or -NRcS02NRc-. Each R6 is independently Rc, halo, -OH, -NH2, -NO2, -CN, or -OCF3. Each Rc is independently hydrogen, an optionally substituted aliphatic, an optionally substituted cycloaliphatic, an optionally substituted heterocycloaliphatic, an optionally substituted aryl, or an optionally substituted heleroaryl. Alternatively, any two adjacent R3 groups together with the atoms to which they are attached form an optionally substituted carbocycle or an optionally substituted heterocycle, or R'3 and an adjacent R3, i.e., attached to the 2 position of the indole of formula la, together with the atoms to which they are attached form an optionally substituted heterocycle.
[00144] In several embodiments, ring B is
[00145] wherein q is 0-3 and each R2o is -ZGR2i , where each ZG is independently a bond or an optionally substituted branched or straightd.s aliphatic chain wherein up to two carbon units of ZG are optionally and independently replaced by -CO-, -CS-, -CONR0-, -C02-, -OCO-, -NRGC02-, -0-, -OCONR0-, -NRGNRG-, -NRGC -, -S-, -SO-, -S02-, -NRG-, -S02NR0-, -NRGS02-, or -NRGS02NRG-. Each R2] is independently RG, halo, -OH, -NH2, -N02, -CN, or -OCF3. Each RG is independently hydrogen, an optionally substituted aliphatic, an optionally substituted cycloaliphatic, an optionally substituted heterocycloaliphatic, an optionally substituted aryl, or an optionally substituted heteroaryl. [001 6] For example, ring 13 is
[00147] In several embodiments, R's is hydrogen and R3 is attached to the 2, 3, 4, 6, or 7 position of the indole of formula la. In several other examples, R3 is attached to the 2 or 3 position of the indole of formula la, and R3 is independently an optionally substituted aliphatic. For instance, R3 is an optionally substituted acyl group. In several instances, R3 is an optionally substituted (alkoxy)carbonyl. In other instances, R3 is (melhoxy)carbonyl, (ethoxy)carbon l, (propoxy)carbonyl, or (butoxy)carbonyl, each of which is optionally substituted with 1 -3 of halo, hydroxy, or combinations thereof. In other instances, R3 is an optionally substituted (aliphatic)carbonyl. For example, R3 is an optionally substituted (alkyl)carbonyl that is optionally substituted with 1 -3 of halo, hydroxy, or combinations thereof . In other examples, R3 is (methyl)carbonyl, (ethyl)carbonyl, (propyl)carbonyl, or (butyl)carbonyl, each of which is optionally substituted with 1 -3 of halo, hydroxy, or combinations thereof.
[00148] In several embodiments, R3 is an optionally substituted (cycloaliphutic)carbonyl or an optionally substituted (heterocycloaliphatic)carbonyl. In several examples, R3 is an optionally substituted (C3-7 cycloaliphatic)carbonyl. For example, R3 is a (cyclopropyl)carbonyl, (cyclobutyl)carbonyl, (cyclopentyl)carbonyl, (cyclohexyl)carbonyl, or (cyc]oheplyl)carbonyl, each of which is optionally substituted with aliphatic, halo, hydroxy, nitro, cyano, or combinations thereof. In several alternative examples, R3 is an optionally substituted (heterocycloaliphatic)carbonyl. For example, R3 is an optionally substituted (helerocycloaliphalic)carbonyl having 1-3 heteroatonis independently selected from N, O, and S. In other examples, R3 is an optionally substituted (heterocycloaliphatic)carbonyl having 1 -3 heteroatonis independently selected from N and O. In still other examples, 3 is an optionally substituted 4-7 membered monocyclic (heterocycloaliphatic)carbonyl having 1 -3 heteroatonis independently selected from N and O. Alternatively, R3 is (piperidine- 1-yl,)carbonyl, (pyrrolidine-l -yl)carbonyl, or (morpholine-4-yl)carbonyl, (piperazine- 1 -yl)carbonyl, each of which is optionally substituted with 1 -3 of halo, hydroxy, cyano, nitro, or aliphatic.
[00149] In still other instances, R3 is optionally substituted (aliphatic)amido such as (aliphatic(amino(carbonyl)) that is attached to the 2 or 3 position on the indole ring of formula la. In some embodiments, R3 is an optionally substituted (alkyl(aniino))carbonyl that is attached to the 2 or 3 position on the indole ring of fonnula la. In other cmbodinienls, R3 is an optionally substituted straight or branched (aliphatic(amino))carbonyl that is attached to the 2 or 3 position on the indole ring of formula la. In several examples, R is (N,N-dimethyl(amino))carbonyl, (methyl(ainino))carbonyl, (elhyl(amino))carbonyl, (propyl(amino))carbonyl, (prop-2-yl(amino))carbonyl, (dimethyl(but-2-yl(amino)))carbonyl, (tertbutyl(amino))carbonyl, (butyl(amino))carbonyl, each of which is optionally substituted with 1 -3 of halo, hydroxy, cycloaliphatic, heterocycloaliphatic, aryl, heteroaryl, or combinations thereof.
[00150] In other embodiments, R3 is an optionally substituted (alkoxy)carbonyl. For example, R3 is (methoxy)carbonyl, (ethoxy)carbonyl, (propoxy)carbonyl, or (butoxy)carbonyl, each of which is optionally substituted with 1 -3 of halo, hydroxy, or combinations thereof. In several instances, R3 is an optionally substituted straight or branched Cj.6 aliphatic. For example, R3 is an optionally substituted straight or branched Ci-6 alkyl. In other examples, R3 is independently an optionally substituted methyl, ethyl, propyl, butyl, isopropyl, or tertbutyl, each of which is optionally substituted with 1 -3 of halo, hydroxy, cyano, nitro, or combination thereof. In other embodiments, R3 is an optionally substituted C3-6 cycloaliphatic. Exemplary embodiments include cyclopropyl, 1 -methyl-cycloprop- l -yl, etc. In other examples, p is 2 and the two R3 substituenls are attached to the indole of formula la at the 2,4- or 2,6- or 2,7- positions. Iixemptary embodiments include 6-F, 3-(optionally substituted Ci^ aliphatic or C3.6 cycloaliphatic); 7-F-2-(-(oplionally substituted Ci.6 aliphatic or C3-6 cycloaliphatic)), 4F-2-(optionally substituted Ct^ ali hatic or cycloaliphatic); 7-CN-2-(optionally substituted Cj-6 aliphatic or C3.6 cycloaliphatic); 7-Me-2-(oplionally substituted C]_6 aliphatic or C3.G cycloaliphatic) and 7-OMc-2-(optionally substituted Cj^ aliphatic or C3-6 cycloaliphatic).
[00151] In several embodiments, R3 is hydrogen.
[00152] In several embodiments, R3 is one selected from: -1-1, -CI-I 3 , -CH2OH, -CH2CH3, -CH2CH2OH, -CH2CH2CH3, -NH2, halo, -OCI I3, -CN, -CI¾, -C(0)0CH2C1-I3, -S(0)2Cl l3, -CI I2NH2, -C(0)NI I2)
[00153] In another embodiment, two adjacent R3 groups form
[00154] In several embodiments, is independently -ZcRe, where each Zc is independently a bond or an optionally substituted branched or straight Ci-6 aliphatic chain wherein up to two carbon units oi Ζ° are optionally and independently replaced by -CO-, -CS-, -CONRc-, -CONRcNRc-, -CO2-, -OCO-, -NRcC02-, -0-, -NRcCONRc-, -OCONRc-, -NRCNRC-, NR CO-, -S-, -SO-, -SO2-, -NR -, -S02NRc-, -NR S02-, or -NRcS02NRc-. Bach R6 is independently Rc, halo, -OH, -NH2, -NO2, -CN, or -OCF3. Each Rc is independently hydrogen, an optionally substituted aliphatic, an optionally substituted cycloaliphatic, an optionally substituted heterocycloaliphatic, or an optionally substituted heteroaryl. In one embodiment, each Rc is hydrogen, Ci-s aliphatic, or C3.6 cycloaliphatic, wherein either of the aliphatic or cycloaliphatic is optionally substituted with up to 4 -OH subslituents. In another embodiment, Rc is hydrogen, or i-e alkyl optionally substituted with up to 4 -OH subslituents.
[00155] For example, in many embodiments, R'3 is is independently -Z Re, where each Z is independently a bond or an optionally substituted branched or straight Ci_6 aliphatic chain wherein up to two carbon units of Z0 are optionally and independently replaced by -C(O)-, -C(0)NRc-, -C(0)0-, -NRcC(0)0-( -0-, -NRcS(0)2-, or -NRC-. Each R6 is independently R c -OH, or -NH2. Each Rc is indepcndenlly hydrogen, an optionally substituted cycloaliphatic, an optionally substituted heterocycloaliphatic, or an optionally substituted heteroaryl. In one embodiment, each Rc is hydrogen, aliphatic, or€3.6 cycloaliphatic, wherein either of the aliphatic or cycloaliphalic is optionally substituted with up to 4 -OH subslituents. In another embodiment, Rc is hydrogen, or Ci^ alkyl optionally substituted with up to 4 -OH subslituents.
[00156] In other embodiments, R'3 is hydrogen or
[00157] wherein R3] is H or a d-2 aliphatic that is optionally substituted with 1 -3 of halo, -OH, or combinations thereof. R.i2 is -L-R33, wherein L is a bond, -Cll2-> -CH20-, -CH2NHS(0)r, -CH2C(0)-, -CH2NHC(0)-, or -CH2NH-; and R33 is hydrogen, or d.2 aliphatic, cycloaliphatic, heterocycloaliphatic, or heteroaryl, each of which is optionally subsitututed with 1 of -OH, -NH2, or -CN. For example, in one embodiment, R31 is hydrogen and R32 is C].2 aliphatic optionally substituted with -OH, -NH2, or -CN.
[00158] In several embodiments, R'3 is independently selected from one of the following:
[00159] In several embodiments, ring B is one selected from: 38 39
[00160] 5. n term
[00161] n is 1 -3.
[00162] In several embodiments, n is 1. In other embodiments, n is 2. In still other embodiments, n is 3.
In one aspect, the present invention relates to compounds of formula 11 aselul as modulators of ABC transporter activity: occurrence: R is H, OH, OCH3 or two R taken together form -OCH20- or -OCF2O-; Ri is H or alkyl; R2 is H or F; R3 is H or CN; R4 is H, -CM 2OC I I2CH (OH)CH 20 I I , -CI^CHjNCQhfc, or -CH2CH2OI I ; R5 is I I, OH, -CH2OCH2CH(OH)CH20H, -CH2OH( or R4 and R5 taken together form a five membered ring.
In one embodiment, the present invention provides compounds of formula II, wherein the compounds set forth below are excluded: In one embodiment of the compounds, two R taken together form -OCF20-, Rt is I I, and R2 is F. In another embodiment, two R taken together form -OCF20-, R] is H, R2 is F, and R3 is H. In another embodiment, two R taken together form -OCF20-, Ri is H, R2 is F, R3 is 11, and R4 is H. In another embodiment, two R taken together form -OCF2O-, Rj is H, R2 is F, R3 is H, and 4 is -CI l2CH2N+(Cl l3) . In another embodiment, two R taken together form -0CF2O, R, is H, R2 is F, R3 is H, and R4 is -CH2OCH2CH(OH)CH20H. In another embodiment, two R taken together form -OCF20-, | is H, R2 is F, R is I I, and R4 and R5 taken together form a five membered ring.
In one embodiment of the compounds, two R taken together form -OCH2O-, Rj is H, and R2 is F. In another embodiment, two R taken together form -OCII20-, Ri is H, R2 is F, and R3 is H. In another embodiment, two R taken together form -OCH20-, Rt is H, R2 is F, R3 is 11, and R4 is -CI I20CI12CJ 1(0H)CI l2OH.
In one embodiment of the compounds, R is OH, Ri is I I, R2 is H, R3 is H, and R4 is -CH2OCH2CH(OH)CH2OH.
In one embodiment of the compounds, at least one R is OCI-I3, at least two Ri arc methyl, R2 is I I, R3 is H, and R4 is H. In another embodiment, at least one R is OCH3, at least two R, are methyl, R2 is II, R3 is I I, and R4 is -CI I2OCH2CII(01 l)CI I2OII.
In one embodiment of the compounds, two R taken together form -CH2CH2CH2-, Ri is H, R2 is H, R3 is H, and R4 is -CH2OCH2CH(OH)CH2OH.
In one embodiment, the compound is represented by formula Ha: WO 2010/054138 PCTYUS200!>/063475 Ila or a pharmaceutically acceptable salt thereof, wherein: R4 is I I, -CH2OCH2CI I(OH)CIJ201 I, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is I I, Oi l, -CH2OCH2CH(01 I)Cl'I2OH, -CH2OI-I, or R4 and R5 taken together form a five membered ring.
[00163] In one embodiment of the compounds, R4 is -CI !2OCH2CH(OH)CH20H, -CI I2CI I2N+(Clli).¾, or -CI I2C1 I2011. In another embodiment, R5 is OM, -CH2OCH2CH(OH)CH20H, or -CH20I 1. In another embodiment, R„ is -CH20CH2C1 I(0H)CI I20H, -CU2CH2N+(CH3)3, or -CH2CH2OH; and R5 is OH, -CH2OCH2CI I(OH)CH2OH, or -CH2OH.
[00164] C. Exemplary compounds of the present invention
[00165] Exemplary compounds of the present invention include, but are not limited to, those illustrated in Table I below.
Table 1 : F.xeniplary compounds of the present invention. 51 52 55 56 58 59 60 61 62 65 66 In another aspect, the present invention relates to a pharmaceutical composition comprising (i) a compound of the present invention; and (ii) a pharmaceutically acceptable carrier. In another embodiment, the composition further comprises an additional agent selected from a mucolytic agent, bronchodialator, an anti-biotic, an anti -infective agent, an anti -inflammatory agent, CFPR corrector, or a nutritional agent. In another embodiment, the composition further comprises an additional agent selected from compounds disclosed in U.S. Patent Application Serial No. 1 1/165,818, published as U.S. Published Patent Application No. 2006/0074075, filed June 24, 2005, and hereby incorporated by reference in its entirety. In another embodiment, the composition further comprises N-(5-hydroxy-2,4-ditert-butyl-phenyl)-4-oxo- l H-quinoline-3-carboxamide. These compositions are useful for treating the diseases described below including cystic fibrosis. These compositions are also useful in the kits described below.
In another aspect, the present invention relates to a method of increasing the number of functional ABC transporters in a membrane of a cell, comprising the step of contacting said ceil with a compound of formula II: II wherein independently for each occurrence: R is I I, OH, OCI l or two R taken together form -CH2CH2CH2-, -OC1120- or -OCH2O-; Ri is H or alkyl; R2 i 1-1 or F; R3 is H or CN; R4 is H, -CH2OCH2CH(OH)CH2OH, -O-hCHzN^CI , or -CH2CH2OH; and R5 is I I, Oi l, -CI I2OCH2CI I(OI I)CI I2OH, -CH2OH, or R4 and R5 taken together form a five membercd ring.
In one embodiment of this method, the ABC transporter is CFrR.
In one embodiment of this method, two R taken together form -OCF20-, Rj is II, and R2 is F. In another embodiment, two R taken together form -OCF20-, R i is H, R2 is F, and R3 is H. In another embodiment, two R taken together form -OCF20-, R] is H, R2 is F, R3 is 11, and R4 is I I. In another embodiment, two R taken together fonn -0CF2O-, Rj is 11, R2 is F, R3 is H, and R4 is -CH2CH2N+(CH3)3. In another embodiment, two R taken together fonn -OCF20-, i is H, R2 is F, R3 is H, and R4 is -CH2OCH2CH(OH)CH2OH. In another embodiment, two R taken together fonn -OCF20-, R] is I I, R2 is F, R3 is H, and R4 and R5 taken together fonn a five membered ring.
In one embodiment of this method, two R taken together fonn -OCH20-, Rj is H, and R2 is F. In another embodiment, two R taken together form -OCH20-, i is H, R2 is F, and R3 is H. In another embodiment, two R taken together fonn -OCTI20-, R | is H, R2 is F, R3 is 11, and R4 is -CH2OCH2CH(OH)CH2OH.
In one embodiment of this method, R is OH, Ri is I I, R2 is H, 3 is II, and R4 is -CH2OCI I2CH(OH)CH2OH.
In one eiiibodiinenl ol' this method, at least one R is OCH3, at least two Ri are methyl, R2 is H, R3 is H, and R is H. In another embodiment, at least one R is OCH3, at least two R i are methyl, R2 is H, R3 is H, and R4 is -CH2OCH2CI l(OH)CH2OH.
In one embodiment of this method, two R taken together form -CH2CH2CH2-, Ri is H, R2 is H, R3 is H, and R is -CH2OCH2CH(OH)CH2OH.
In one embodiment of this method, the compound is represented by formula Ila: Ila or a pharmaceutically acceptable salt thereof, wherein: R4 is I I, -CH2OCI-I2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH ; and R5 is I I , OH, -CH2OCH2CH(OH)CI-l2OH, -CH2OH, or 4 and R taken together form a five membered ring.
In one embodiment of this method, 4 is -CH2OCI I2CI I(OU)CU2OI I, -CH2CI I2N+(CH3)3, or -CI I2C1 I20H. In another embodiment, R5 is OH, -CH20CH2C1 I(0H)C1 I201I, or -CH2OI 1. In another embodiment, R4 is -CH2OCH2CH(OH)CH2OI I, -CH2CH2N+(CH3)3, or -CH2CI I2OH; and R5 is OH, -CH2()C:H2CH(C)H)CI-I2()H, or -CH2OH.
In one embodiment of this method, the compound is selected from Table 1.
In another aspect, the present invention relates to a method of treating a condition, disease, or disorder in a patient implicated by ABC transporter activity, comprising the step of administering to said patient a compound having formula II: II or ii pharmaceutically acceptable salt thereof, wherein independently for each occurrence: R is H, OH, OCH3 or two R taken together form -CH2CI l2CH2-, -OCH20- or -OCF20-; Rj is 11 or alkyl; R2 i s I I or F; R3 is H or CN; R4 is I I, -CH20CH2CH(OH)CH20H, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is I I, OH, -CHiOCH2CH(OH)CH20H, -CH2OH, or R4 and R5 taken together form a five memhercd ring.
Ln one embodiment of this method, two R taken together form -OCF20-, | is H, and R2 is J7. Γη another embodiment, two R taken together form -OCF20-, ] is 11, R2 is I7, and R3 is H. In another embodiment, two R taken together fonn -OCF20-, Rj is H, R2 is F, R3 is H, and R4 is H. In another embodiment, two R taken together form -OCF20-, R j is I I , R2 is F, R3 is II, and R4 is -CH2CI-l2N+(CH3)3. In another embodiment, two R taken together fonn -OCF2O-, R, is H, R2 is F, R3 is H, and R4 is -CII2OCH2C] !(OH)CH2OH. In another embodiment, two R taken together fonn -OCF20-, Ri is 11, R2 is F, R3 is H, and R4 and R5 taken together form a five membered ring.
In one embodiment of this method, two R taken together fonn -OCH20-, Rj is H, and R2 is F. In another embodiment, two R taken together form -OCH20-, Rt is H, R2 is F, and R3 is H. In another embodiment, two R taken together fonn -OCH2O-, R| is H, R2 is F, R3 is H, and R4 is -CH2OCH2CH(OH)CH2OH.
In one embodiment of this method, R is Oi l, Ri is H, R2 is H, 3 is H, and R4 is -CH2OCH2CH(OH)CH2OH.
In one embodiment of this method, at least one R is OCI I3, at least two Ri are methyl, R2 is I I, R3 is II, and R4 is I I. In another embodiment, at least one R is OCII3, at least two Ri are methyl, R2 is H, R3 is H, and R4 is -CH2OCI-I2CH(01-I)CH2OH.
In one embodiment of this method, two R taken together fon -CH2CH2CH2-, Ri is 11, R2 is H, R3 is H, and R4 is -CH20CH2CH(OH)CH2OH.
In one embodiment of this method, the compound is represented by formula I la: l la or a pharmaceutically acceptable salt thereof, wherein: ¾ is I I , -CH2OCH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is I I , OH, -Cl l2OCI l2CH(01 l)CI l2OII, -C1 I20H, or R4 and R5 taken together form a five niembered ring.
In one embodiment of this method, R4 is -CH2OCH2CH( H)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH. In another embodiment, R5 is OH, -CH2OCH2CI l(OH)CH2OH, or -CH2OH. In another embodiment, R4 is -CH2OCH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is OH, -CI l2OCH2CH(OH)CI l20H, or -CH2OH.
In one embodiment of this method, the compound is selected from Table 1.
In one embodiment of this method, said condition, disease, or disorder is selected from cystic fibrosis, hereditary emphysema, hereditary hemochromatosis, coagulalion-fibrinolysis deficiencies, such as protein C deficiency, Type 1 hereditary angioedema, lipid processing deficiencies, such as familial hypercholesterolemia, Type 1 chyloniicronemia, abetalipoproteinemia, lysosomal storage diseases, such as I-cell disease/pseudo-Hurler, mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II, polyendocrinopathy hyperi nsulemia, diabetes mellitus, laron dwarfism, myleoperoxidase deficiency, primary hypoparathyroidism, melanoma, glycanosis CDG type 1 , hereditary emphysema, congenital hyperthyroidism, osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency, diabetes insipidus (di), neurophyseal di, neprogenic Dl, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson' s disease, amyotrophic lateral sclerosis, progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders asuch as Huntington, spinocerebullar ataxia type I, spinal and bulbar muscular atrophy, dcntatorubal pallidoluysian, and myotonic dystrophy, as well as spongiform encephalopathies, such as hereditary Creulzfeldt-Jakob disease, Fabry disease, Straussler-Scheinker syndrome, COPD, dry-eye disease, and Sjogren's disease.
In another aspect, the present invention relates to a kit for use in measuri ng the activity of a ABC transporter or a fragment thereof in a biological sample in vitro or in vivo, comprising: (i) a first composition comprising a compound of formula II: II wherein independently for each occurrence: R is H, OH, OCH3 or t wo R taken together form -CHzCH2CH2-, -OCH20- or -OCF20-; R i is H or alkyl; R2 i H or F; R-, is I I or CN; R, is II, -CII2OCI l2Cl l(OI l)CH2OII, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is II, OH, -CH2OCH2CH(OH)CH2OH, -CH2OH, or R4 and R5 taken together form a five menibered ring; and (ii) instructions for: a) contacting the composition with the biological sample; and b) measuring activity of said ABC transporter or a fragment thereof.
In one embodiment, the kit further comprises instructions for a) contacting an additional composition with the biological sample; b) measuring the activity of said ABC transporter or a fragment thereof in the presence of said additional compound, and c) comparing the activity of the ABC transporter in the presence of the additional compound with the density of the ABC transporter in the presence of said first composition.
In one embodiment, the kit is used to measure the density of CFfR.
In one embodiment of this kit, two R taken together form -OCF20-, Ri is H, and R2 is F. In another embodiment, two R taken together form -OCF20-, R| is 1-1, R2 is F, and R3 is H. In another embodiment, two R taken together form -OCF20-, R] is H, R2 is F, R3 is H, and R4 is H. In another embodiment, two R taken together form -OCF2O-, Ri is H, R2 is F, Rj is H, and R4 is -CH2CH2N+(CH3)3. In another embodiment, two R taken together form - 0CF2O-, R ) is H, R2 is F, R3 is H, and R4 is -CH2OCH2CH(OH)CH2OH. In another embodiment, two R taken together form -OCF20-, R] is H, R2 is F, R3 is H, and R4 and R5 taken together form a five membered ring, In one embodiment of this kit, two R taken together form -OCH2O-, Ri is H, and R2 is F. In another embodiment, two R taken together form -OCH20-, R i is H, R2 is F, and R3 is H. In another embodiment, two R taken together form -OCH20-, R i is H, R2 is F, R3 is H, and R4 is -CH2OCH2CI I(OI-I)CH2OH. In another embodiment, R is OH, , is H, R2 is H, R3 is H, and is -CH20CH2CH(OH)CH2OM. In another embodiment, at least one R is OCH3, at least two R i are methyl, R2 is H, R3 is I I, and R4 is I I. In another embodiment, at least one R is OCH3, at least two R i are methyl, R2 is I I, R3 is H, and R4 i s -CH2OCH2CH(OH)CH2OH. In another embodiment, two R taken together form -CH2CH2CH2-, Ri is I I, R2 is I I, R3 is H, and R4 is -CI^OCr^CI OIDCI ^OI I.
In one embodiment of this kit, the compound is represented by formula lla: Ila or a pharmaceutically acceptable salt thereof, wherein: R4 is I I, -CH2OCH2CH(OH)CH20H, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is 11, OH, -CH2OCH2CI ICOH)CH2OH, -CH20I1, or R4 and R5 taken together form a five membered ring.
In one embodiment of this kit, R is - H2OCH2CH(OH)CH20H, -CH2CH2N+(CH3)3, or -CH2CH2OH. In another embodiment, R5 is OH, -CH20CH2CH(OH)CH2OH, or -CH2OH. In another embodiment, R4 is -CH2OCH2CH(OH)CH20H, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is OH, -CH20CH2CH(0H)CH2OH, or -CH2OH.
In one embodiment of this kit, the compound is selected from Table 1.
[00166] HI. SUBGENFRIC COMPOUNDS OF THE PRESENT INVENTION
[00167] Another aspect of the present invention provides a compound that is useful for modulating ABC transporter activity. 'I"he compound has formula Id: Ic or a pharmaceutically acceptable salt thereof.
[00168] Ri, R2, and ring A are defined above in formula 1, and ring B, R3 and p are defined in fomiula la. Furthermore, when ring A is unsubstituted cyclopentyl, n is 1 , R2 is 4-chloro, and Ri is hydrogen, then ring B is not 2-(tertbutyl)indol-5-yl, or (2,6-dichlorophenyl(carbonyl))-3-methyl- lH-indol-5-yl; and when ring A is unsubstituted cyclopentyl, n is 0, and R, is hydrogen, then ring B is not.
[00169] Another aspect of the present invention provides a compound that is useful for modulating ABC transporter activity. The compound has formula Id: Id or a pharmaceutically acceptable salt thereof.
[00170] R), R2, and ring A are defined above in formula I, and ring B, R3 and p are defined in fomiula la.
[00171] However, when Ri is I I, n is 0, ring A is an unsubstituted cyclopentyl, and ring B is an indole-5-yl substituted with 1-2 of R3, then each R3 is independently -ZaRi2, where each 7? is independently a bond or an unsubstituted branched or straight Ci^ aliphatic chain wherein up to two carbon units of ZG are optionally and independently replaced by -CS-, -CONRGNRG-, -CO2-, -OCO-, -NRGC02-, -0-, -NRGCONRG-, -OCONR0-, -NRGNRG-, -S-, -SO-, -SO2-, -NRG-, -S02NRG-, -NRGS02-, or -NR S02NRc-, each R12 is independently R°, halo, -OH, -NH2, -N02, -CN, or -OCF3, and each RG is independently hydrogen, an unsubstituted aliphatic, an optionally substituted cycloaliphalic, an optionally substituted heterocycloaliphatic, an unsubstituted aryl, or an optionally substituted heteroaryl; or any two adjacent R3 groups together with the atoms to which they are attached form an optionally substituted hctcrocycle. Fuihemiore, when Rj is H, n is 1 , R2 is 4-chloro, ring A is an unsubstituted cyclopenlyl, and ring B is an indole-5-yl substituted with 1-2 of R3, then each R3 is independently -ZHR22> where each ZH is independently a bond or an unsubstitutcd branched or straight Cj.3 aliphatic chain wherein up to two carbon units of ZH are optionally and independently replaced by -CS-, -CONRHNR", -CO2-, -OCO-, -NRHC02-, -0-, -NRHCONR"-, -OCONRH-, -NRHNRh-, -S-, -SO-, -SO2-, -NRH-, -S02NR -, -NRHS02-, or -NRHS02NRH-, each R22 is independently R1!, halo, -OH, -NH2) -N02, -CN, or -OCF3, and each R11 is independently hydrogen, a substituted C4 alkyl, an optionally substituted C2-6 alkenyl, an optionally substituted C2 alkynyl, an optionally substituted C4 alkenyl, an optionally substituted C alkynyl, an optionally substituted cycloaliphalic, an optionally substituted heterocycloaliphatic, an optionally substituted heteroaryl, an unsubstituted phenyl, or a mono-substituted phenyl, or any two adjacent 3 groups together with the atoms to which they are attached form an optionally substituted heterocycle.
[00172] Another aspect of the present invention provides a compound that is useful for modulating ABC transporter activity. The compound has formula II: II or a pharmaceutically acceptable salt thereof.
[00173] Rj , R2, and ring A are defined above in formula I; R3, R'3, and p are defined above in formula la; and Z Z2, Z3, Z4, and Z5 are defined above in formula lb.
[00174] Another aspect of the present invention provides a compound that is useful for modulating ABC transporter activity. rV e compound has formula Ila: Ha or a pharmaceutically acceptable salt thereof.
[00175] R|, R2, and ring A are defined above in formula I; R3, R'3, and p are defined above in formula la; and Zi, Z2, Z3, Z , and Z5 are defined above in formula lb.
[00176] Another aspect of the present invention provides a compound that is useful for modulating ABC transporter activity. The compound has formula lib: lib or a pharmaceutically acceptable salt thereof.
[00177] R), R2, and ring A, are defined above in formula I; R3, R 3, and p are defined above in fomiula la; and Zj, Z2, Zi, ¾, and Z5 arc defined above in formula lb.
[00178] Another aspect of the present invention provides a compound that is useful for modulating ABC transporter activity. The compound has formula lie: lie or a pharmaceutically acceptable salt thereof.
[00179] R], R2 and n are defined above in formula I; and R3) R'3) and p are defined in formula la.
[00180] Another aspect of the present invention provides a compound that is useful for modulating ABC transporter activity. rl"he compound has fomiula lid: or a pharmaceutically acceptable salt thereof.
[00181] Both R2 groups, together with the atoms to which they are attached form a group selected from: XA19 XA20 XA21
[00182] R'3 is independently selected from one of the following: and each R3 is independently selected from -H, -CH3, -CH2OH, -CH2CH3, -CH2CH2OH, -CH2CH2CH3, -NH2, halo, -O JCUHIj3,, --CI:N,, --CL*F33., •C(0)oci i2cu3, -s(0)2cn3, -CH2NII2, -C(0)NH2,
[00183] IV. GENERIC SYNTHETIC SCHEMES
[00184] ITie compounds of fonnulae (I, Ic, Id, II, Ila, lib, lie, and lid) may be readily synthesized from commercially available or known starting materials by known methods. Exemplary synthetic routes to produce compounds of formulae (I, Ic, Id, II, Ila, lib, lie, and lid) are provided below in Schemes 1-22 below.
[00185] Preparation of the compounds of the invention is achieved by the coupling of a ring B amine with a ring Λ earboxylic acid as illustrated in Scheme 1 .
[00186] Scheme 1 : DCM DMF.
[00187] Referring Lo Scheme 1 , the acid 1 a may be converted to the corresponding acid chloride lb using thionyl chloride in the presence of a catalystic amount of dimethylformamidc. Reaction of the acid chloride with the amine Ri N~\J provides compounds of the invention I. Alternatively, the acid l may be directly coupled to the amine using known coupling reagents such as, for example, HATU in the presence of triethylamine.
[00188] Preparation of the acids la may be achieved as illustrated in Scheme 2.
Scheme 2: a) NaOII, BTEAC; b) NaOH, Δ
[00189] Referring to Scheme 2, the nitrile 2a reacts with a suitable bromochloroalkane in the presence of sodium hydroxide and a phase tranfer catalyst such as butyltriethylammonium chloride to provide the intermediate 2b. Hydrolysis of the nitrilc of 2b provides the acid l a. In some instances, isolation of the intermediate 2b is unnecessary.
[00190] The phenylacetonitriles 2a are commercially available or may be prepared as illustrated in Scheme 3.
Scheme 3 3d 2a a) Pd(PPh3)„, CO, MeOH; b) LiAlH4, THF; c) SOCl2; d) NaCN
[00191] Referring to Scheme 3, reaction of an aryl bromide 3a with carbon monoxide in presence of methanol and telrakis(triphenylphosphine)palladium (0) provides the ester 3b.
Reduction of 3b with lithium aluminum hydride provides the alcohol 3c which is converted to the halide 3d with thionyl chloride. Reaction of 3d with sodium cyanide provides the nitrite 2a.
[00192] Other methods of producing the nitrile 2a are illustrated in schemes 4 and 5 below. Scheme 4 a) TosMIC; b) NaBH4, THF; c) SOCl2; d) NaCN Scheme 5 5a 5b 2a a) NBS, AIBN, CCU; b) NaCN, ElOH
[00193] l^reparalion of components is illustrated in the schemes that follow.
A number of methods for preparing ring B compounds wherein ring B is an indole have been reported. See for example Angew. Chem. 2005, 44, 606; J . Am. Chem. Soc. 2005, 127, 5342,); J. Comb. Chem. 2005, 7, 130; Tetrahedron 2006, 62, 3439; J. Chem. Soc. Perkin Trans. 1, 2000, 1045. 11 _ "-\
[00194] One method (or preparing Rr' -^jj^ is i llustrated in Scheme 6.
Scheme 6 a) NaN02, HC1, SnCl2; b) NaOI I, R3CH2C(0)R3, EtOH; c) Ι Ι3ΙΌ4, toluene; d) H2, Pd-C, EtOH
[00195] Referring to Scheme 6, a nitroaniline 6a is converted Lo the hydrazine 6b using nitrous acid in the presence of HC1 and stannous chloride. Reaction of 6b with an aldehyde or ketone CH3C(0)R3 provides the hydrazone 6c which on treatment with phophoric acid in toluene leads to a mixture of nitro indoles 6d and 6e. Catalytic hydrogenalion in the presence of palladium on carbon provides a mixture of the amino indoles 6f and 6g which may be separated using know methods such as, for example, chromatography.
[00196] An alternative method is illustrated in scheme 7.
[00197] Scheme 7 a) R3aCOCl, I¾N, CU2C12; b) n-BuLi, TI IF; c) NaBl , AcOII; d) KN03, H2S04; e) DDQ, 1 ,4-dioxane; f) NaN02, HCl, SnCl2.2H20, H20; g) eCOR3, EtOH; h) PPA; i) Pd/C, ElOH or H2, Raney Ni, EtOH or MeOH
[00198]
[00199] Scheme 8 a) HN03, H2S04; b) Me2NCH(OMc)2, DMF; c) H2, Rancy Ni, EtOH Scheme 9 a) HN03, I I2SO4; b) SOCl2; EtOll; c) DMA, DMF; d) Raney Ni, H2, MeOII
[00202] Scheme 11 a) DMA, DMF; b) Raney Ni, H2, MeOH
[00203] Scheme 12 a) R3llCH2C0R3b, AcOH, ElOH; b) H3P0 , toluene; c) H2, Pd/C, EtOH
[00204] Scheme 14 a) NaliH3CN; b) When PG= S()2Ph: PhS02Cl, Et3N, DMAP, C112C12; When PG= Ac: AcCI, NaI IC03l CH2C12; c) When R = RCO: (RCO)20, A1C13( CH2C12; When Rv=Br: Br2, AcOH; d) I I Br or HCl; e) KN03, H2S04; f) Mn02, CH2a2 or DDQ, 1 ,4-dioxane; g) H2, Raney Ni, EtOH.
[00205] Scheme 14 a) NaBH3CN; b) RS02C1, DMAP, Et3N, CH2C12; c) RDC(0)CI, A1C13, CH2C12; d) NaBRi, THF; e) HBr; 0 KN03, H2S02; g) Mn()2; g) Raney Ni, H2, EtOH
[00206] Scheme 15 a) R3X (X=Br, I), zinc Lriflate, TBAI, DIEA, toluene; b) 1-12, Rancy Ni, EtOH or H2) Pd/C, EtOH or SnCl2.2H20, EtOH; c) C1S02NC0, DMF, CH3CN
[00207] Scheme 16 a) when X=C1, Br, I, or OTs: R'3X, K2C03, DMF or CH3CN; b) H2, Pd/C, EtOH or SnCl2.2l l20, EtOH or SnCl2.2H20, DIEA, EtOH.
[00208] Scheme 17 a) Br2, AcOM; b) RC(0)C1, I¾N, CH2C12; c) HC≡€R3a, Pd(PPh3) 2Cl2, Cul, El3N; d) TBAF, TI-IF or tBuOK, DMF or Pd(PPh3) 2Cl2, Cul, DMF; e) H2, Pd/C, EtOH or SnCl2, MeOH or HCO2NH4, Pd C, EtOH
[00209] Scheme 18 a) Br2, AcOH, CIIC13; b) R3aCaCH, Cul, I¾N, Pd(PPh3) 2Cl2; c) RCOC1, I¾N, CH2C12; d) TBAF, DMF; e) Rancy Ni, H2, MeOH; f ROK, DMF
[00210] Scheme 19 a) 13r2, AcOH; b) HC=CR3n, Pd(PPh3) 2C12, Cul, I¾N; c) Pd(PPh3)2Cl2, Cul, DMF; d) H2, Pd/C, BtOH or SnCl2, MeOH or HC02NH4, Pd/C, EtOH
[00211] Scheme 20 a) H2NR'3; b) X=Br: Br2, HOAc; X=I: NIS; c) HC≡CR3, Pd(PPh3)2Cl2, Cul, Et3N; d) Cul, DMF or TBAF, THF; e) 112, Pd/C, EtOH or SnCI2, MeOH or HC02NH4, Pd C, EtOH
[00212] Scheme 21 a) R'3NH2, DMSO; b) Br2, AcOH; c) TMS-C≡CH, Cul, TEA, Pd(PPh3) 2Cl2; d) Cul, DMSO; e) Raney Ni, H2, MeOH
[00213] Scheme 22 a) R¾C=CH, Cut, TEA, Pd(PPh3) 2C12; b) TBAF, THF; c) Raney Ni, MeOH
[00214] Scheme 23 a) NaBH4, NiCl2, MeOH; b) RC(0)C1; c) Pd(PPh3)Cl2, HG≡C-R3, Cul, ¾N; d) IBuOK, DMF; e) N0 , H2S04; f) NaB¾, NiCI2, MeOH
[00215] Scheme 24 a) SnCl2, ElOH or Pd/C, HCO2NH4 or H2, Pd/C, EtOH or Raney Ni, H2, ElOH
[00216] Scheme 25 a) PPh3, HBr; h) Cl(0)CCH2C02Et; c) tBuOK; d) (Boc) 20, DMAP; e) KHMDS, R-X; KHMDS, R-X; f) TFA; g) NaNC , H2S04; h) LiAlH4, THE; i) SnCl2, EtOH
[00217] Scheme 26 a) LiOH; b) EDC, HOBt, Et3N, HNRyRz; c) BH3-THF; d) if Rz=H, RC(0)C1 (Z=RC(0)-) or RS02C1 (S^RSOi-) or RO(CO)Cl (Z=RO(CO)-) or (RO(CO)) 20 (Z= Z=RO(CO)-). 1¾N, CH2C12
[00218] Scheme 27 a) R'3-X (X=Br, I, or Ol s), base (K2C03 or Cs2C03), DMF or CU3CN; b) H2, Pd/C, EtOH or Pd/C, HCO2NH4
[00219] Scheme 28 a) R3aX (X=C1, Br, I), A1C13, CH2C12; b) Rancy Ni, H2, McOH
[00220] Scheme 29 a) HCl/MeOH; Pl02, H2; b) (Boc) 20, Et3N, THF
[00221] Scheme 30 a) NaOl-1 or LiOl l; b) ROl l, 11C1; c) NaBH4 or LiAlIL- or D1BAL-H, I'HF; d) HNRyRz, HATU, EtaN, ELOH or DMF; e) LiAlI , THF or I3H3 THF; f) H202, H20 (Ry=Rz=Il); g) H2, Pd/C
[00222] Scheme 31 a) R0-X, NaH; Rb-X, Nai l; b) PC15, CH2CI2; c) NaOH; d) NaNH2) DMSO; e) CH2N2; 0 Pd(PPh3)4, Cul, Et3N; g) RC(0)C1, pyr, CH2C12; h) Pd(CH3CN) 2C12, CH3CN; i) Raney Ni, H2, MeOH
[00223] Scheme 32 a) LiOH, THF H20; b) I INRyRz, HATU, TEA, DMF/CH2Cl2
[00224] Scheme 33 a) LiBH4, THF/H20 or LiAlH4, THF; b) Ra-Li, THF
[00225] Scheme 34 a) NaBH3CN; b) R'3CHO, NaHB(OAc) 3. TFA, DCE; c) chloranil or CDCI3, light or DDQ
[00227] Scheme 36 a) NBS; b) Ar-B(OR)2, Pd-FibreCat 1007, K2CO„ EtOH
[00229] Scheme 38 a) RSO2CI, NaH, THF-DMF; b) R3-X (X=Br, i, or O'l's), NaH, THF-DMF; c) ethylene dioxide, Ιη(¾; d) POCl3, DM1'; e) H2N-OH, CH2C12; Ac20
[00230] Scheme 39 a) NaH, THF-DMF; epirfilorohydrin; b) ROH; c) HNRyR:
[00231] Scheme 40 a) TsCl, Et3N, CH2C12; b) NaCN, DMF; c) NaOH, MeOH; d) NaN3, NH4C1; e) NaN3, DMF; 0 Pd/C, H2, MeOH (R=H); h) RxC(0)CI (Z=RxC(0)-) or RxSOzCl (Z=R*S02-) or R*0(C0)Cl (Z=R*0(CO)-) or (R"0(CO)) 20 ( Z=RxO(CO)-), Ei3N, CH2C12
[00232] Scheme 41 a) C1CH2CH0, NaHB(OAc)3, CH2C12; CDC13, light; b) NaN3, Nal, DMF; c) H2, IWC, MeOH, AcOH; d) RC(0)C1 (Z=RC(0)-) or RSO.Cl (Z=RS02-) or RO(CO)CI (Z=RO(CO)-) or (RO(CO)) 20 (Z= RO(CO)-), Et3N, CH2C12.
Scheme 42 b) Ra-X, Nai l; Rh-X, NaH; b) PC15, CH2C12; c) NaOH; d) NaNH2, DMSO; e) R-OH, DCC; 0 PdCPPh3)2Cl2, Cul, }¾N; g) PdCl2, CIl3CN Scheme 43 n= 0 or 1 a) DIBAL-H; b) P-LG; P= proLecLing group like TBDMS and LG= leaving group like CI; c) R4-LG, base likeCs2C03; R4 is alkyl and LG is tosylate, Rc=H or R4; d) reducing conditions like Pd/C, Hz or ammonium formate.
Scheme 43 R4-LG, base likeCs2C03; R4 is alkyl and LG is tosylate; b) L1AIH4; c) reducing conditions like Pd/C, H2 or ammonium formate.
[00233] In the schemes above, the radical R employed therein is a substituent, e.g., RW as defined hereinabove. One of skill in the art will readily appreciate that synthetic routes suitable for various substituents of the present invention are such that the reaction conditions and steps employed do not modify the intended substituents.
[00234] V. FORMULATIONS, ADMINISTRATIONS, AND USES
[00235] Accordingly, in another aspect of the present invention, pharmaceutically acceptable compositions are provided, wherein these compositions comprise any of the compounds as described herein, and optionally comprise a pharmaceutically acceptable carrier, adjuvant or vehicle. In certain embodiments, these compositions optionally further comprise one or more additional therapeutic agents.
[00236] It will also be appreciated that certain of the compounds of present invention can exist in free form for treatment, or where appropriate, as a pharmaceutically acceptable derivative or a prodrug thereof. According to the present invention, a pharmaceutically acceptable derivative or a prodrug includes, but is not limited to, pharmaceutically acceptable salts, esters, salts of such esters, or any other adducl or derivative which upon administration to a patient in need is capable of providing, directly or indirectly, a compound as otherwise described herein, or a metabolite or residue thereof.
[00237] As used herein, the term "pharmaceutically acceptable salt" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. A "pharmaceutically acceptable salt" means any non-toxic salt or salt of an ester of a compound of this invention that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention or an inhibitorily active metabolite or residue thereof.
[00238] Pharmaceutically acceptable salts are well known in the art. For example, S. . Berge, et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1 77, 66, 1 - 19, incorporated herein by relerence. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrale, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hcmisulfate, heptanoatc, hcxanoate, hydroiodidc, 2-hydroxy-ethanesulfonatc, lactobionale, lactate, laurale, lauryl sulfate, malale, maleate, malonate, methanesulfonate, 2-naphthalcnesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phcnylpropionatc, phosphate, picrate, pivalate, propionate, stcarate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(Ci-4alky!)4 salts. This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersible products may be obtained by such quaternization. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.
[00239] As described above, the pharmaceutically acceptable compositions of the present invention additionally comprise a pharmaceutically acceptable carrier, adjuvant, or vehicle, which, as used herein, includes any and all solvents, diluents, or other liquid vehicle, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired. Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., ( 980) discloses various carriers used in formulating pharmaceutically acceptable compositions and known techniques for the preparation thereof. Except insofar as any conventional carrier medium is incompatible with the compounds of the invention, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other componenl(s) of the pharmaceutically acceptable composition, its use is contemplated to be within the scope of this invention. Some examples of materials which can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, or potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymeihyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols; such a propylene glycol or polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulalor.
[00240] In yet another aspect, the present invention provides a method of treating a condition, disease, or disorder implicated by ABC transporter activity. In certain embodiments, the present invention provides a method of treating a condition, disease, or disorder implicated by a deficiency of ABC transporter activity, the method comprising administering a composition comprising a compound of formulae (I, Ic, Id, II, Ila, lib, lie, and lid) to a subject, preferably a mammal, in need thereof.
[00241] In certain preferred embodiments, the present invention provides a method of treating Cystic fibrosis, Hereditary emphysema, Hereditary hemochromatosis, Coagulation-Fibrinolysis deficiencies, such as Protein C deficiency, Type 1 hereditary angioedema, Lipid processing deficiencies, such as Familial hypercholesterolemia, Ί ype 1 chylomicronemia, Abctalipoproteincmia, Lysosomal storage diseases, such as I-cell disease/Pscudo-Hurlcr, Mucopolysaccharidoses, Sandhof/Tay-Saehs, Crigler-Najjar type II, Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus, Laron dwarfism, Myleoperoxidase deficiency, Primary hypoparathyroidism, Melanoma, Glycanosis CDG type 1 , Hereditary emphysema, Congenital hyperthyroidism, Osteogenesis imperfecta, Hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI), Neurophyseal DI, Neprogenic DI, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, Progressive supranuclear plasy, Pick's disease, several polygluiamine neurological disorders asuch as Huntington, S inocerebullar ataxia type I, Spinal and bulbar muscular atrophy, Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as Spongiform encephalopathies, such as Hereditary Creutzfeldt-Jakob disease (due to Prion protein processing defect), Fabry disease, Straussler-Scheinker disease, secretory diarrhea, polycystic kidney disease, chronic obstructive pulmonary disease (COPD), dry eye disease, and Sjogren's Syndrome, comprising the step of administering to said mammal an effective amount of a composition comprising a compound of formulae (I, Ic, Id, II, Ila, lib, lie, and lid), or a preferred embodiment thereof as set forth above.
[00242] According Lo an alternative preferred embodiment, the present invention provides a method of treating cystic fibrosis comprising the step of administering to said mammal a composition comprising the step of administering to said mammal an effective amount of a composition comprising a compound of formulae (I, lc, Id, II, JIa, lib, lie, and lid), or a preferred embodiment thereof as set forth above.
[00243] According to the invention an "effective amount" of the compound or pharmaceutically acceptable composition is that amount effective for treating or lessening the severity of one or more of Cystic fibrosis, Hereditary emphysema, Hereditary hemochromatosis, Coagulation-Fibrinolysis deficiencies, such as Protein C deficiency, Type 1 hereditary angioedema, Lipid processing deficiencies, such as Famiiial hypercholesterolemia, Type 1 chylomicronemia, Abetalipoproteinemia, Lysosomal storage diseases, such as I-cell disease/Pseudo- Hurler, Mucopolysaccharidoses, Sandhof Tay-Sachs, Crigler-Najjar type II, Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus, Laron dwarfism, Myleoperoxidase deficiency, Primary hypoparathyroidism, Melanoma, Glycanosis CDG type 1, Hereditary emphysema, Congenital hyperthyroidism, Osteogenesis imperfecta, Hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI), Neurophyseal DI, Neprogenic DI, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, Progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders asuch as Huntington, Spinocerebullar ataxia type I, Spinal and bulbar muscular atrophy, Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as Spongiform encephalopathies, such as Hereditary Creutz eldt-Jakob disease, Fabry disease, Straus sler-Scheinker disease, secretory diarrhea, polycystic kidney disease, chronic obstructive pulmonary disease (COPD), dry eye disease, and Sjogren's Syndrome.
[00244] The compounds and compositions, according to the method of the present invention, may be administered using any amount and any route of administration effective for treating or lessening the severity of one or more of Cystic fibrosi , Hereditary emphysema, Hereditary hemochromatosis, Coagulation-Fibrinolysis deficiencies, such as Protein C deficiency, Type 1 hereditary angioedema, Lipid processing deficiencies, such as Familial hypercholesterolemia, Type 1 chylomicronemia, Abetalipoproteinemia, Lysosomal storage diseases, such as I-cell disease/Pseudo-Hurler, Mucopolysaccharidoses, Sandhof Tay-Sachs, Crigler-Najjar type II, Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus, Laron dwarfism, Myleoperoxidase deficiency, Primary hypoparathyroidism, Melanoma, Glycanosis CDG type 1, Hereditary emphysema, Congenital hyperthyroidism, Osteogenesis imperfecta, Hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI), Neurophyseal DI, Neprogenic DI, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, Progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders asuch as Huntington, Spinocercbullar ataxia type I, Spinal and bulbar muscular atrophy, Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as Spongiform encephalopathies, such as Hereditary Creutzfeldt-Jakob disease, Fabry disease, Slraussler-Scheinker disease, secretory diarrhea, polycystic kidney disease, chronic obstructive pulmonary disease (COPD), dry eye disease, and Sjogren's Syndrome.
[00245] The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the particular agent, its mode of administration, and the like. The compounds of the invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The expression "dosage unit form" as used herein refers to a physically discrete unit of agent appropriate for the patient to be treated. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific effective dose level for any particular patient or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed, and like factors well known in the medical arts. The term "patient", as used herein, means an animal, preferably a mammal, and most preferably a human.
[00246] The pharmaceutically acceptable compositions of this invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), bucally, as an oral or nasal spray, or (he like, depending on the severity of the ini'ection being treated. In certain embodiments, the compounds of the invention may be administered orally or parenterally at dosage levels of about 0.01 mg kg to about 50 mg/kg and preferably from about 1 mg kg to about 25 mg kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.
[00247] Liquid dosage I onus for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microeinulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, genii, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
[00248] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be fonnulated according to (he known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S. P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.
[00249] The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
[00250] In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. I ¾e rate of absorption of the compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound fonn is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylaclide-polyglycolide. Depending upon the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoestcrs) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.
[00251] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
[00252] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable cxcipicnt or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannilol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, poly vinylpyrrolidinonc, sucrose, and acacia, c) hunicctants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol nionostearale, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.
[00253] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coalings and other coalings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredients) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.
[00254] The active compounds can also be in microencapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coalings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.
[00255] Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, eardrops, and eye drops are also contemplated as being within the scope of this invention. Additionally, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms are prepared by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rale controlling membrane or by dispersing the compound in a polymer matrix or gel.
[00256] As described generally above, the compounds of the invention are useful as modulators of ABC transporters. Thus, without wishing to be bound by any particular theory, the compounds and compositions are particularly useful for treating or lessening the severity of a disease, condition, or disorder where hyperactivity or inactivity of ADC transporters is implicated in the disease, condition, or disorder. When hyperactivity or inactivity of an ABC transporter is implicated in a particular disease, condition, or disorder, the disease, condition, or disorder may also be referred to as a "ABC transporter-medi ted disease, condition or disorder". Accordingly, in another aspect, the present invention provides a method for treating or lessening the severity of a disease, condition, or disorder where hyperactivity or inactivity of an ABC transporter is implicated in the disease slate.
[00257] The activity of a compound utilized in this invention as a modulator of an ABC transporter may be assayed according to methods described generally in the art and in the Examples herein.
[00258] It will also be appreciated that the compounds and pharmaceutically acceptable compositions of the present invention can be employed in combination therapies, that is, the compounds and pharmaceutically acceptable compositions can be administered concurrently with, prior to, or subsequent to, one or more other desired therapeutics or medical procedures. The particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved. It will also be appreciated that the therapies employed may achieve a desired effect for the same disorder (for example, an inventive compound may be administered concurrently with another agent used to treat the same disorder), or they may achieve different effects (e.g., control of any adverse effects). As used herein, additional therapeutic agents that are normally administered to treat or prevent a particular disease, or condition, are known as "appropriate for the disease, or condition, being treated".
[00259] The amount of additional therapeutic agent present in the compositions of this invention will be no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent. Preferably the amount of additional therapeutic agent in the presently disclosed compositions will range from about 50% to 100% of the amount normally present in a composition comprising that agent as the only therapeutically active agent.
[00260] The compounds of this invention or pharmaceutically acceptable compositions thereof may also be incorporated into compositions for coating an implantable medical device, such as prostheses, artificial valves, vascular grafts, stents and catheters.
Accordingly, the present invention, in another aspect, includes a composition for coating an implantable device comprising a compound of the present invention as described generally above, and in classes and subclasses herein, and a carrier suitable for coating said implantable device. In still another aspect, the present invention includes an implantable device coated with a composition comprising a compound of the present invention as described generally above, and in classes and subclasses herein, and a carrier suitable lor coaling said implantable device. Suitable coatings and the general preparation of coated implantable devices are described in US Patents 6,099,562; 5,886,026; and 5,304, 121. The coatings are typically biocompatible polymeric materials such as a hydrogel polymer, polymelhyldisiloxanc, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate, and mixtures thereof. The coatings may optionally be further covered by a suitable topcoat of fluorosilicone, polysaccarides, polyethylene glycol, phospholipids or combinations thereof to impart controlled release characteristics in the composition.
[00261] Another aspect of the invention relates to modulating ABC transporter activity in a biological sample or a patient (e.g., in vitro or in vivo), which method comprises administering to the patient, or contacting said biological sample with a compound of formula I or a composition comprising said compound. The term "biological sample", as used herein, includes, without limitation, cell cultures or extracts thereof; biopsied material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.
[00262] Modulation of ABC transporter activity in a biological sample is useful for a variety of purposes that are known to one of skill in the art. Examples of such purposes include, but are not limited to, the study of ABC transporters in biological and pathological phenomena; and the comparative evaluation of new modulators of ABC transporters.
[00263] In yet another embodiment, a method of modulating activity of an anion channel in vitro or in vivo, is provided comprising the step of contacting said channel with a compound of fonnulae (I, Ic, Id, II, lla, lib, lie, and lid). In preferred embodiments, the anion channel is a chloride channel or a bicarbonate channel. In other preferred embodiments, the anion channel is a chloride channel.
[00264] According to an alternative embodiment, the present invention provides a method of increasing the number of functional ABC transporters in a membrane of a cell, comprising the step of contacting said cell with a compound of fonnulae (I, lc, Id, II, lla, lib, lie, and Ild). The term "functional ABC transporter" as used herein means an ABC transporter that is capable of transport activity. In preferred embodiments, said functional ABC transporter is CFTR.
[00265] According to another preferred embodiment, the activity of the ABC transporter is measured by measuring the transmembrane voltage potential. Means for measuring the voltage potential across a membrane in the biological sample may employ any of the known methods in the art, such as optical membrane potential assay or other electrophysiological methods.
[00266] The optical membrane potential assay utilizes voltage-sensitive FRET sensors described by Gonzalez and Tsien (See, Gonzalez, J. E. and R. Y. Tsien ( 1995) "Voltage sensing by fluorescence resonance energy transfer in single cells" Biophys J 69(4): 1272-80, and Gonzalez, J. E. and R. Y. Tsien (1 97) "Improved indicators of cell membrane potential that use fluorescence resonance energy transfer" Chem Biol 4(4): 269-77) in combination with instrumentation for measuring fluorescence changes such as the Voltage/Ion Probe Reader (V[PR) (See, Gonzalez, J. E„ K. Oades, et al. (1999) "Cell-based assays and instrumentation for screening ion-channel targets" Drug Discov Today 4(9): 431-439).
[00267] These voltage sensitive assays are based on the change in fluorescence resonant energy transfer (FRET) between the membrane-soluble, voltage-sensitive dye, DiSBAC2(3), and a fluorescent phospholipid, CC2-DMPE, which is attached to the outer leaflet of the plasma membrane and acts as a FRET donor. Changes in membrane potential (Vm) cause the negatively charged iSBAC2(3) to redistribute across the plasma membrane and the amount of energy transfer from CC2-DMPE changes accordingly. The changes in fluorescence emission can be monitored using VIPR™ II, which is an integrated liquid handler and fluorescent detector designed to conduct cell-based screens in 96- or 384-well niicrotiter plates.
[00268] In another aspect the present invention provides a kit for use in measuring the activity of a ABC transporter or a fragment thereof in a biological sample in vitro or in vivo comprising (i) a composition comprising a compound of formulae (1, Ic, Id, II, Ila, lib, lie, and lid) or any of the above embodiments; and (ii) instructions for a.) contacting the composition with the biological sample and b.) measuring activity of said ABC transporter or a fragment thereof. In one embodiment, the kit further comprises instructions for a.) contacting an additional composition with the biological sample; b.) measuring the activity of said ABC transporter or a fragment thereof in the presence of said additional compound, and c.) comparing the activity of the ABC transporter in the presence of the additional compound with the density of the ABC transporter in the presence of a composition of formulae (1, lc, Id, II, Ila, lib, lie, and lid). In preferred embodiments, the kit is used to measure the density of CFTR.
[00269] In order that the invention described herein may be more fully understood, the following examples are set forth. should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention in any manner.
[00270] VI. PREPARATIONS AND EXAMPLES
[00271] General Procedure 1: Carboxylic Acid Building Block Hal = CI, Br, I
[00272] Benzyltriethylammonium chloride (0.025 equivalents) and the appropriate dihalo compound (2.5 equivalents) were added to a substituted phenyl acetonitrile. The mixture was heated at 70 °C and then 50 % sodium hydroxide (10 equivalents) was slowly added lo the mixture. The reaction was stirred at 70 °C for 12-24 hours to ensure complete formation of the cycloalkyl moiely and then heated at 130 DC for 24-48 hours lo ensure complete conversion from the nitrilc to the carboxylic acid. ITic dark brown / black reaction mixture was diluted with water and extracted with dichloromethane three times to remove side products. The basic aqueous solution was acidified with concentrated hydrochloric acid to pH less than one and the precipitate which began to form at pH 4 was filtered and washed with 1 M hydrochloric acid two times. The solid material was dissolved in dichloromethane and extracted two times with 1 M hydrochloric acid and one time with a saturated aqueous solution of sodium chloride. The organic solution was dried over sodium sulfate and evaporated lo dryness to give the eycloalkylcarbox lic acid. Yields and purities were typically greater than 90%.
[00273] Example 1: l-Benzo[l,3]dioxol-5-yl-cyclopropanecarboxylic acid
[00274] A mixture of 2-(benzold]ll,3jdioxol-5-yl)acetonitrile (5.10 g 31.7 mmol), 1 -bromo-2-chloro-ethane (9.00 niL 109 mmol), and benzyltriethylammonium chloride (0.181 g, 0.795 mmol) was heated at 70 °C and then 50% (wt./wt.) aqueous sodium hydroxide (26 niL) was slowly added to the mixture. The reaction was stirred at 70 "C for 24 hours and then healed at 130 ° for 48 hours. The dark brown reaction mixture was diluted with water (400 mL) and extracted once with an equal volume of ethyl acetate and once with an equal volume of dichloromethane. The basic aqueous solution was acidified with concentrated hydrochloric acid to pll less than one and the precipitate filtered and washed with 1 hydrochloric acid. The solid material was dissolved in dichloromethane (400 mL) and extracted twice with equal volumes of 1 M hydrochloric acid and once wilh a saturated aqueous solution of sodium chloride. The organic solution was dried over sodium sulfate and evaporated to dryness to give a white to slightly off-white solid (5.23 g, 80%) ESI-MS tn/z calc. 206.1 , found 207.1 ( + l )+. Retention lime 2.37 minutes. !H NMR (400 MHz, DMSO-d6) fi 1.07- 1. 1 1 (m, 2H), 1.3S- I .42 (ra, 2H), 5.98 (s, 2H), 6.79 (m, 2H), 6.88 (m, 1 H), 12.26 (s, 1 H).
[00275] General Procedure II: Carboxylic Acid Building Block Hal = CI, Br, I, all other variables are as defined in the text.
[00276] Sodium hydroxide (50 % aqueous solution, 7.4 equivalents) was slowly added to a mixture of the appropriate phenyl acetonitrile, bcnzyltrieihylammonium chloride ( 1.1 equivalents), and the appropriate dihalo compound (2.3 equivalents) at 70 °C. The mixture was stirred overnight at 70 °C and the reaction mixture was diluted with water (30 mL) and extracted wilh ethyl acetate. The combined organic layers were dried over sodium sulfate and evaporated to dryness to give the crude cyclopropanecarbonitrile, which was used directly in the next step.
[00277] The crude cyclopropanecarbonitrile was refluxed in 10% aqueous sodium hydroxide (7.4 equivalents) for 2.5 hours. The cooled reaction mixture was washed with ether ( 100 mL) and the aqueous phase was acidified to pH 2 with 2M hydrochloric acid. The precipitated solid was filtered to give the cyclopropanecarboxylic acid as a white solid.
[00278] General Procedure III: Carboxylic Acid Building Block Example 2: l-(2,2-Difluoro-benzo[l,3]dioxol-5-yl)-cyclopropanecarboxylic
[00280] 2,2-Difluoro-benzo[l,3]dioxole-5-carboxy]ic acid methyl ester
[00281] A solution of 5-bromo-2,2-difluoro-benzo| . l ,3]dioxole (1 1.8 g, 50.0 mmol) and tetrakis(triphenylphosphine)palladiuni (0) [Pd(PPh3)4, 5.78 g, 5.00 mmol] in methanol (20 mL) containing acctoniirilc (30 mL) and tricthy!aminc ( 10 mL) was stirred under a carbon monoxide atmosphere (55 PSI) at 75 °C (oil bath temperature) for 15 hours. Ί Tie cooled reaction mixture was filtered and the filtrate was evaporated to dryness. The residue was purified by silica gel column chromatography to give crude 2,2-difluoro-benzo 1 1 ,3] dioxole-5-carboxylic acid methyl ester (11.5 g), which was used directly in the next step.
[00282] (2,2-Dilliioro-benzo[] ,3]dioxo)-5-y])-methanol
[00283] Crude 2,2-difluoro-benzo| 1 ,3 ]dioxole-5-carboxylic acid methyl ester ( 1 1 .5 g) dissolved in 20 mL of anhydrous letrahydrofuran (TI IF) was slowly added to a suspension of lithium aluminum hydride (4.10 g, 106 mmol) in anhydrous THF ( 100 mL) at 0 °C. The mixture was then warmed to room lenipcrature. After being stirred at room temperature for I hour, the reaction mixture was cooled to 0 °C and treated with water (4.1 g), followed by sodium hydroxide (10% aqueous solution, 4.1 mL). ITie resulting slurry was filtered and washed with THF. The combined filtrate was evaporated to dryness and the residue was purified by silica gel column chromatography to give (2,2-difluoro-benzo[ l ,3]dioxol-5-yl)-methanol (7.2 g, 38 mmol, 76 % over two steps) as a colorless oil.
[00284] 5-Chloromethyl-2,2-difluoro-benzo[ 1 ,3]dioxole
[00285] Thionyl chloride (45 g, 38 mmol) was slowly added to a solution of (2,2-dtfluoro-benzoj 1 ,3]dioxo!-5-yl)-methanol (7.2 g, 38 mmol) in dichloromethane (200 mL) at 0 °C. The resulting mixture was stirred overnight at room temperature and then evaporated to dryness. The residue was partitioned between an aqueous solution of saturated sodium bicarbonate ( 100 mL) and dichloromethane ( 100 mL). The separated aqueous layer was extracted with dichloromethane (150 mL) and the organic layer was dried over sodium sulfate, filtrated, and evaporated to dryness to give crude 5-chloromelhyl-2,2-difluoro-benzo[l ,3]dioxole (4.4 g) which was used directly in the next step.
[00286] (2,2-Difluoro-benzolL3]dioxoLS yl)-acelonitrile
[00287] A mixture of crude 5-chloromethyl-2,2-difluoro-benzo| 1 ,3]dioxole (4.4 g) and sodium cyanide (1.36 g, 27.8 mmol) in dimethylsulfoxide (50 mL) was stirred at room temperature overnight. The reaction mixture was poured into ice and extracted with ethyl acetate (300 mL). The organic layer was dried over sodium sulfate and evaporated to dryness to give crude (2,2-difiuoro-benzo[ l ,3 |dioxol-5-yl)-acetonitrile (3.3 g) which was used directly in the next step.
[00288] 1 -(2,2-DifIuoro-benzof 1 ,3]dioxol-5-y l)-cyclopropanecarbonitrile
[00289] Sodium hydroxide (50% aqueous solution, 10 mL) was slowly added to a mixture of crude (2,2-dinuoro-benzoLl ,3 |dioxol-5-y])-acetoniLrile, ben yltrieLhylammonium chloride (3.00 g, 15.3 mmol), and 1 -bromo-2-chloroethane (4.9 g, 38 mmol) at 70 °C.
[00290] The mixture was stirred overnight at 70 °C before the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate and evaporated to dryness to give crude l-(2,2-difluoro-benzo| 1 ,3]dioxol-5-yl)-cyclopropanecarbonitrile, which was used directly in the next step.
[00291] l-(2,2-Difluoro-benzo[l,3]dioxol-5-yl)-cyclopropanecarboxylic acid
[00292] l -(2)2-Di luoro-benzo[ l ,3 idioxol-5-yl)-cyclopropanecarbonitrile (crude from the last step) was refluxed in 10% aqueous sodium hydroxide (50 mL) for 2.5 hours. The cooled reaction mixture was washed with ether ( 100 mL) and the aqueous phase was acidified to pH 2 with 2M hydrochloric acid. The precipitated solid was filtered to give l -(2,2-difluoro-benzo| 1 ,3]dioxol-5-yl)-cyclopropanecarboxylic acid as a while solid (0.15 g, 1.6% over four steps). liSI-MS m/z calc. 242.04, found 241.58 (M+ l)+; !H NMR (CDC13) δ 7.14-7.04 (m, 2H), 6.98-6.96 (m, 111), 1.74- J .64 (m, 211), 1.26- 1.08 (m, 2H).
[00293] Example 3: 2-(2,2-Dimethylbenzo[d][l,3]dioxol-5-yl)acetonitrile ^ ^ \_#*^0
[00294] (3,4-Di h droxy- pheny t)-acetonit rile
[00295] To a solution of benzol 1 ,3 ldioxol-5-yl-acetonitrile (0.50 g, 3.1 mmol) in CH2C12 (15 mL) was added dropwise Brir3 (0.78 g, 3.1 mmol) at -78 °C under N2. The mixture was slowly warmed to room temperature and stirred overnight. H20 (10 mL) was added to quench the reaction and the CH2C12 layer was separated. The aqueous phase was extracted with CH2CI2 (2 x 7 ml ,). The combined organics were washed with brine, dried over Na2S04 and purified by column chromatography on silica gel (petroleum ether/ethyl acetate 5: 1) to give (3,4-dihydroxy-pheny])-acetonitri]e (0.25 g, 54%) as a white solid. Ή NMR (DMSO-rfe, 400 MHz) δ 9.07 (s, 1 I I), 8.95 (s, 1 II), 6.68-6.70 (m, 2 I I), 6.55 (dd, J = 8.0, 2.0 Hz, I I I), 3.32 (s, 2 H).
[00296] 2-(2,2-Dimethylbenzotd][l,3]dioxol-5-y])acetonitrile
[00297] ΓΓο a solution of (3,4-dihydroxy-phenyl)-acelonitrile (0.20 g, 1.3 mmol) in toluene (4 mL) was added 2,2-dimethoxy-propane (0.28 g, 2.6 mmol) and TsOH (0.010 g, 0.065 mmol). The mixture was heated at reflux overnight. The reaction mixture was evaporated to remove the solvent and the residue was dissolved in ethyl acetate. The organic layer was washed with NallCO^ solution, H20, brine, and dried over Na2SC>4. The solvent was evaporated under reduced pressure to give a residue, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 10: 1 ) to give 2-(2,2-dimethylbenzo[d]l l ,3jdioxol-5-yl)acetonitrile (40 mg, 20%). Ί-1 NMR (CDC13, 400 MHz) δ 6.68-6.71 (m, 3 H), 3.64 (s, 2 H), 1 .67 (s, 6 H).
[00298] Example 4: l-(3,4-Dihydr xy-phcnyl)-cyclopropanecarboxylic acid
[00299] l-(3,4-Bis-benzyloxy-phenyl)-cyclopropanecarbonitrile
[00300] To a mixture of (n-C4Hg)4NBr (0.50 g, 1.5 mmol), toluene (7 mL) and (3,4-bis-benzyloxy-phenyl)-acelonilrile (14 g, 42 mmol) in NaOH (50 g) and H20 (50 mL) was added BrCH2CHzCl (30 g, 0.21 mol). The reaction mixture was stirred at 50 °C for 5 h before being cooled to room temperature. Toluene (30 mL) was added and the organic layer was separated and washed with H20, brine, dried over anhydrous MgS04, and concentrated. 'Hie residue was purified by column on silica gel (petroleum ether/ethyl acetate 10: 1 ) to give I-(3,4-bis-benzyloxy-phenyl)-cyclopropanecarbonitrile ( 10 g, 66%). ]H NMR (DMSO 300 MHz) δ 7.46-7.30 (m, 10 H), 7.03 (d, ./ = 8.4 Hz, 1 H), 6.94 (d, J = 2.4 H/„ I H), 6.89 (dd, J Hz, 1 I I), 5.12 (d, J = 7.5 Hz, 411), 1.66- 1.62 (m, 2 H), 1.42- 1.37 (m, 2 H). [00301 ] l-(3,4-Dihydroxy-phenyl)-cyclopropanecarbonitrile
[00302] To a solution of 1 -(3,4-bis-benzyloxy-phcnyl)-cye]opropanecarbonitrilc ( 10 g, 28 nimol) in MeOH (50 niL) was added Pd/C (0.5 g) under nitrogen atmosphere. 'Hie mixture was stirred under hydrogen atmosphere ( 1 atm) at room temperature for 4 h. The catalyst was filtered off through a celite pad and the filtrate was evaporated under vacuum to give 1 -(3,4-dihydroxy-phenyO-cyclopropanecarbonitrile (4.5 g, 92%). Ή NMR (D SO 400 MHz) δ 9.06 (br s, 2 11), 6.67-6.71 (m, 2 II), 6.54 (dd, J = 2.4, 8.4 Hz, 1 II), 1.60- 1 .57 (m, 2 H), 1.30- 1.27 (m, 2 H).
[00303] l-(3,4-Dihydroxy-phenyl)-cyclopropanecarboxylic acid
[00304] To a solution of NaOH (20 g, 0.50 mol) in H2() (20 mL) was added I -(3,4-dihydroxy-pheny -cyclopropanecarbonitrile (4.4 g, 25 nimol). The mixture was heated at reflux for 3 h before being cooled to room temperature. The mixture was neutralized with HCl (0.5 N) lo pH 3-4 and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with water, brine, dried over anhydrous MgS04, and concentrated under vacuum to obtain l-(3,4-dihydroxy-phenyl)-cyclopropanecarboxylic acid (4.5 g crude). From 900 nig crude, 500 mg pure l-(3,4-dihydroxy-phenyl)-cyclopropanecarboxylic acid was obtained by preparatory HPLC Ή NMR (DMSO.300 MHz) δ 12.09 (br s, 1 H), 8.75 (br s, 2 H), 6.50-6.67 (m, 3 H), 1.35-1.31 (m, 2 1-1), 1.01-0.97 (m, 2 H).
[00305] Example 5: 1-(2-Oxo-2,3-dihydrobenzo[d]oxazol-5-yl)cyclopropane-car boxy lie acid.
[00306] 1 -(4- ethoxy-phenyl)-cyclopropanecarboxylic acid methyl ester
[00307] To a solution of 1 -(4-me oxy-phenyl)-cyclopropanecarboxylic acid (50 g, 0.26 mol) in McOl l (500 inL) was added to!ucnc-4-suli'onic acid monohydratc (2.5 g, 13 minol) ai room temperature, 'l c reaction mixture was heated at reflux for 20 hours. MeOH was removed by evaporation under vacuum and EtOAc (200 mL) was added. The organic layer was washed with sat. aq. Nal lC03 ( 100 mL) and brine, dried over anhydrous Na2S04 and evaporated under vacuum to give l -(4-methoxy-phenyl)-cyclopropanecarboxylic acid methyl ester (53 g, 99%). Ή NMR (CDCl.i.400 MHz) δ 7.25-7.27 (m, 2 H), 6.85 (d, J = 8.8 Hz, 2 H), 3.80 (s, 3 H), 3.62 (s, 3 H), 1 .58 (q, J = 3.6 Hz, 2 H), 1.15 (q, J = 3.6 Hz, 2 H).
[00308] l-(4-Methoxy-3-nitro-phenyI)-cyclopropanecarboxylic acid methyl ester
[00309] To a solution of l -(4-melhoxy-phenyl)-cyclopropanecarboxylic acid methyl ester (30.0 g, 146 mmo!) in AczO (300 mL) was added a solution of HN03 (I4.1 g, 146 mmol, 65%) in AcOH (75 mL) at 0 °C. The reaction mixture was stirred at 0 ~ 5 °C for 3 h before aq. HCI (20%) was added dropwise at 0 °C. The resulting mixture was extracted with EtOAc (200 inL x 3). The organic layer was washed with sat. aq. NaHCC>3 then brine, dried over anhydrous Na2S04 and evaporated under vacuum to give l-(4-methoxy-3-nitro-phenyl)-cyclopropanecarboxylic acid methyl ester (36.0 g, 98%), which was directly used in the next step. Ή NMR (CDC13.300 MHz) δ 7.84 (d, J = 2.1 Hz, 1 H), 7.54 (dd, 7 = 2.1 , 8.7 Hz, i H), 7.05 (d, J = 8.7 Hz, 1 H), 3.97 (s, 3 H), 3.65 (s, 3 H), 1.68-1.64 (m, 2 11), 1 .22- 1. 18 (m, 2 I I).
[00310] l-(4-Hydroxy-3-nitro-phenyl)-cyclopropanecarboxylic acid methyl ester
[00311] To a solution of 1 -(4-methoxy-3-nitro-phenyl)-cyclopropane-carboxylic acid methyl ester ( 10.0 g, 39.8 mniol) in CH2C12 (100 mL) was added BBr3 ( 12.0 g, 47.8 mmul) ut. -70 °C. The mixture was stirred at -70 °C for 1 hour, then allowed lo warm to -30 °C and stirred at this temperature for 3 hours. Water (50 mL) was added drop wise at -20 °C, and the resulting mixture was allowed to warm room temperature before it was extracted with ElOAc (200 mL x 3). Ί Tie combined organic layers were dried over anhydrous Na2S0 and evaporated under vacuum to give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 15: 1 ) to afford l -(4-hydroxy-3-nitro-phenyl)-cyclopropanecarboxylic acid methyl ester (8.3 g, 78%). Ή NMR (CDC , 400 Hz) δ 10.5 (s, I H), 8.05 (d, ,/ = 2.4 Hz, 1 H), 7.59 (dd, J = 2.0, 8.8 Hz, I H), 7.1 1 (d, J = 8.4 Hz, 1 H), 3.64 (s, 3 H), 1.68- 1.64 (in, 2 H), 1.20-1.15 (m, 2 H).
[00312] l-(3-Amino-4-hydroxy-phenyl)-cyclopropanecarboxylic acid methyl ester
[00313] To a solution of 1 -(4-hydroxy-3-nitro-phenyl)-cyclopropanccarboxylic acid methyl ester (8.3 g, 35 mniol) in eOH ( 100 mL) was added Raney Nickel (0.8 g) under nitrogen atmosphere. ITie mixture was stirred under hydrogen atmosphere (1 aim) at 35 °C for 8 hours. The catalyst was filtered off through a Celite pad and the iiltrate was evaporated under vacuum to give crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 1 : 1 ) to give l-(3-amino-4-hydroxy-phcnyl)-cyclopropanecarboxylic acid methyl ester (5.3 g, 74%). Ή NMR (CDC13, 400 MHz) δ 6.77 (s, 1 I-I), 6.64 (d, J s 2.0 Hz, 2 H), 3.64 (s, 3 H), 1.55- 1.52 (m, 2 H), 1.15- 1.12 (m, 2 H).
[00314] l-(2-Oxo-2,3-dihydro-benzooxazol-5-yl)-cyclopropanecarboxylic acid methyl ester
[00315] To a solution of l-(3-amino-4-hydroxy-phenyl)-cyclopropanecarboxylic acid methyl ester (2.0 g, 9.6 nmiol) in THF (40 mL) wtis added triphosgene (4.2 g, 1 nimol) at room temperature. The mixture was stirred for 20 minutes at this temperature before water (20 mL) was added dropwise al 0 °C. The resulting mixture was extracted with EtOAc (100 mL x 3). The combined organic layers were dried over anhydrous Na2S04 and evaporated under vacuum to give 1 -(2-oxo-2,3-dihydro-benzooxazol-5-yl)-cyclopropanecarboxylic acid methyl ester (2.0 g, 1%), which was directly used in the next step. Ή NMR (CDCI3.300 MHz) δ 8.66 (s, 1 H), 7.1 3-7.12 (m, 2 H), 7.07 (s, I H), 3.66 (s, 3 H), 1.68- 1.65 (m, 2 H), 1 .24- 1.20 (m, 2 H). [003Ϊ6] l-(2-Oxo-2,3-dihydrobenzotd]oxazol-5-yl)cyc.opropanecarboxylic acid
[00317] To a solution of 1 -(2-oxo-2,3-dihydro-bcnzooxazol-5-yl)-cyclopropanecarboxylic acid methyl ester ( 1.9 g, 8.1 mniol) in MeOH (20 mL) and water (2 mL) was added LiOH.I- O (1.7 g, 41 nimol) in portions at room temperature. The reaction mixture was stirred for 20 hours at 50 °C. MeOH was removed by evaporation under vacuum before water ( 100 mL) and BtOAc (50 mL) were added. The aqueous layer was separated, acidified with HC1 (3 mol/L) and extracted with BtOAc (100 mL x 3). The combined organic layers were dried over anhydrous a2S04 and evaporated under vacuum to give I -(2-oxo-2,3-dihydrobenzo|dJoxazol-5-yl)cyclopropanecarboxylic acid ( 1.5 g, 84%). Ή NMR (DMSO. 400 MHz) δ 12.32 (brs, 1 H), 1 1.59 (bra, 1 H), 7.16 (d, J = 8.4 Hz, 1 11), 7.00 (d, = 8.0 Hz, 1 H), 1 .44- 1 .41 (m, 2 H), 1. 13- 1. 10 (m, 2 H). MS (LSI) m/e (M+H+) 218.1.
[00318] Example 6: l-(6-Fluoro-b nzo[l,3]dioxol-5-yl)-cyclopropanecarboxylic acid
[00319] 2-Fluoro-4,5-dihydroxy-benzaldehyde
[00320] To a stirred suspension of 2-fluoro-4,5-dimethoxy-benzaldehyde (3.00 g, 16.3 mmol) in dichloromethane (100 niL) was added BBr3 ( 1.2.2 mL, 130 ramol) dropwise at -78 °C under nitrogen atmosphere. After addition, the mixture was wanned to -30 °C and stirred at this temperature for 5 h. The reaction mixture was poured into ice water and the precipitated solid was collected by filtration and washed with dichloromethane to afford 2-fluoro-4,5-dihydroxy-benzaldehyde (8.0 g), which was used directly in the next step.
[00321] 6-riuoro-ben-O[l,3]dioxol€-5-carbaldehyde To a stirred solution of 2-fluoro-4,5-dihydroxy-benzaldehyde (8.0 g) and BrClCH2 (24.8 g, 1 0 mmol) in dry DMP (50 mL) was added Cs2CC>3 (62.0 g, 1 0 mmol) in portions. The resulting mixture was stirred at 60 °C overnight and then poured into water. 'ITie mixture was extracted with EtOAc (200 mL x 3). The combined organic layers were washed with brine (200 mL), dried over Na2S04, and evaporated in vacuo to give crude product, which was purified by column chromatography on silica gel (5-20% ethyl acetate/petroleum ether) to afford 6-fluoro-benzo| l ,3Jdioxo!e-5-carbaldehydc (700 mg, two steps yield: 24%). Ί ΐ-NMR (400 MHz, CDC13) δ 10.19 (s, 1 I I), 7.23 (d, J = 5.6, 1 I I), 6.63 (d, J = 9.6, 1 H), 6.08 (s, 2 H).
[00322] (6-Fluoro-benzo[l,3]dioxol-5-yl)-methanoI
[00323] To a stirred solution of 6-fluoro- benzol 1 ,3]dioxole-5-carbaldehyde (700 mg, 4.2 mmol) in MeOH (50 mL) was added NaBH4 (320 mg, 8.4 mmol) in portions at 0 °C. The mixture was stirred at this temperature for 30 min and was then concentrated in vacuo to give a residue. rI¾e residue was dissolved in EtOAc and the organic layer was washed with water, dried over Na2SC>4, and concentrated in vacuo to afford (6-fluoro-benzo[l ,3]dioxol-5-yl)-methanol (650 mg, 92%), which was direcdy used in the next step.
[00324] 5-Chloromethyl-6- 1uoro-benzo[l,3]dioxole
[00325] (6-Fluoro-benzo[ 1 ,3 ]dioxol-5-yl)-methanol (650 mg, 3.8 mmol) was added to SOCI2 (20 niL) in portions at 0 °C. ri¾e mixture was warmed lo room temperature for 1 h and then heated at reflux lor 1 h. The excess SOCI2 was evaporated under reduced pressure to give the crude product, which was basified with sat. Nal CO.i solution to pl l ~ 7. The aqueous phase was extracted with EtOAc (50 mL x 3). The combined organic layers were dried over Na2S04 and evaporated under reduced pressure to give 5-chloromethyl-6-fluoro-benzo[l ,3]dioxole (640 mg, 90%), which was directly used in the next step.
[00326] (6-Fluoro-benzo[l ,3]dioxol-5- vlVacctonitrile
[00327] A mixture of 5-chloromethyl-6-fluoro-benzo[ l ,3]dioxo1e (640 mg, 3.4 mmol) and NaCN (340 mg, 6.8 mmol) in DMSO (20 mL) was stirred at 30 °C for 1 h and then poured into water. The mixture was extracted with EtOAc (50 mL x 3). The combined organic layers were washed with water (50 mL) and brine (50 mL), dried over NaaSC^, and evaporated under reduced pressure to give the crude product, which was purified by column chromatography on silica gel (5-10% ethyl acetate/petroleum ether) to afford (6-fluoro-benzo[ l ,3]dioxol-5-yl)-acetonitrile (530 mg, 70%). Ή-N R (300 MHz, CDC13) δ 6.82 (d, ,/ = 4.8, ί H), 6.62 J = 5.4, 1 H). 5.99 (s, 2 H), 3.65 (s, 2 H).
[00328] 1 - (6-F1 uoro-benzo[l ,3]dioxol -5- y l)-cyclopropanecarbonitri le
[00329] A flask was charged with water ( 10 mL), followed by a rapid addition of NaOH (10 g, 0.25 mol) in three portions over a 5 min period. The mixture was allowed lo cool lo room temperature. Subsequently, the flask was charged with toluene (6 mL), tclrabutyl-ammonium bromide (50 mg, 0.12 mmol), (6-fluoro-benzoLl,3|dioxol-5-yl)-acelonitrile (600 mg, 3.4 mmol) and l-bromo-2-chloroethane (1.7 g, 12 mmol). The mixture stirred vigorously at 50 °C overnight. The cooled flask was charged with additional toluene (20 mL). The organic layer was separated and washed with water (30 mL) and brine (30 mL). rITie organic layer was removed in vacuo to give the crude product, which was purified by column chromatography on silica gel (5- 10% ethyl acetate/petroleum elher) to give l-(6-nuoro-benzo[ l ,3|dioxol-5-yl)-cyclopropanecarbonitrile (400 mg, 60%). Ή NMR (300 MHz, CDCl^) δ 6.73 (d, ./ = 3.0 Hz, 1 H), 6.61 (d, ./ = 9.3 Hz, 1 H), 5.98 (s, 2 II), i .67- 1.62 (m, 2 H), 1.31- 1.27 (m, 2 H).
[00330] 1 -(6-Fluoro-benzo[l ,3]dioxol-5-yl)-cyclopropanecarboxylic acid
[00331] A mixture of l -(6-nuoro-benzo[ l,3Jdioxol-5-yl)-cyclopropanccarbonitrile (400 mg, 0.1 6 nimol) and 10% NaOH ( 10 mL) was stirred at 100 °C overnight. After the reaction was cooled, 5% HC1 was added until the pH < 5 and then ElOAc (30 mL) was added to the reaction mixture. The layers were separated and combined organic layers were evaporated in vacuo to afford l -(6-fluoro-benzo[ l ,3 |dioxol-5-yl)-cyclopropanecarboxylic acid (330 mg, 76%). Ή NMR (400 MHz, DMSO) 8 12.2 (s, 1 H), 6.87-6.85 (m, 2 H), 6.00 (s, 1 H), 1.42-1.40 (m, 2 H), 1.14- 1 .07 (m, 2 I I).
[00332] Example 7: l-(Benzofuran-5-yl)cyclopropanecarboxylic acid
[00333] l-[4-(2,2-Diethoxy-ethoxy)-phenyl]-cyclopropanecarboxylic acid
[00334] To a stirred solution of l -(4-hydroxy-phenyl)-cyclopropanecarboxylic acid methyl ester ( 15.0 g, 84.3 mmol) in DMF (50 mL) was added sodium hydride (6.7 g, 170 nimol, 60% in mineral oil) at 0 DC. After hydrogen evolution ceased, 2-bromo- l, l-diethoxy-ethane (16.5 g, 84.3 mmol) was added dropwise to the reaction mixture. The reaction was slimed at 160 X for 15 hours. The reaction mixture was poured onto ice ( 100 g) and was extracted with CH2CI2. The combined organics were dried over Na2S04. rI e solvent was evaporated under vacuum to give l -[4-(2,2-diethoxy-ethoxy)-phenyl )-cyclopropanecarboxylic acid ( 10 g), which was used directly in the next step without purification.
[00335] l-Benzofuran-5-yl-cyclopropanecarboxylic acid
[00336] To a suspension of 1 -[4-(2,2-dielhoxy-ethoxy)-phcnyl]-cyclopropaneearboxylic acid (20 g, -65 mmol) in xylene (100 niL) was added PPA (22.2 g, 64.9 mmol) at room temperature. The mixture was heated at reflux ( 140 °C) for 1 hour before it was cooled to room temperature and decanted from the ΡΡΛ. The solvent was evaporated under vacuum to obtain the crude product, which was purified by preparative HPLC to provide 1 -(benzofuran-5-yl)cyclopropanccarboxylic acid ( 1.5 g, 5%). *H NMR (400 MHz, DMSO-4f) δ 12.25 (br s, 1 H), 7.95 (d, ./ = 2.8 Hz, 1 I I), 7.56 (d, ./ = 2.0 Hz, 1 H), 7.47 (d, ./ = 1 1.6 Hz, 1 H), 7.25 (dd, J = 2.4, 1 1.2 Hz, 1 H), 6.89 (d, J = 1.6 Hz, 1 H), 1.47- 1.44 (m, 2 H), 1.17- 1.14 (m, 2 H).
[00337] Example 8: l-(2,3-Dihydrobenzofuran-6-yl)cyclopropanecarboxylic acid
[00338] To a solution of l -(benzofuran-6-yl)cyclopropanecarboxylic acid (370 nig, 1.8 mmol) in MeOH (50 ml.,) was added Pt02 (75 mg, 20%) at room temperature. The reaction mixture was stirred under hydrogen atmosphere ( 1 atni) at 20 °C for 3 d. The reaction mixture was filtered and the solvent was evaporated in vacuo to afford the crude product, which was purified by prepared HPLC to give l-(2,3-dihydrobenzofuran-6-yl)cyclopropanecarboxylic acid (155 mg, 42%). Ή NMR (300 MHz, MeOD) δ 7.13 (d, J = 7.5 Hz, 1 H), 6.83 (d, J = 7.8 Hz, 1 H), 6.74 (s, 1 H), 4.55 (t, J = 8.7 Hz, 2 H), 3.18 (t, ./ = 8.7 Hz, 2 H), 1.56- 1 .53 (m, 2 I I), 1.19-1.15 (m, 2 H).
[00339] Example 9: H3,3-Dimethyl-2,3-dihydrobenzofuran-5-ylicy clopropanecarboxylic acid.
[00340] 1 -(4-Hydroxy-phenyl)-cyclopropanecarhoxylic acid methyl ester
[00341] To a solution of methyl l -(4-methoxyphenyl)cyclopropanecarboxylate (10.0 g, 48.5 mmol) in dichloromcthane (80 niL) was added 1¾SI I ( 16 mL) under ice-water bath. The mixture was stirred at 0 °C for 20 min before A1C13 ( 1 .5 g, 0.15 mmol) was added slowly at 0 °C The mixture was stirred at 0 °C for 30 min. The reaction mixture was poured into ice-watcr, the organic layer was separated, and the aqueous phase was extracted with dichloromethane (50 mL x 3). ' he combined organic layers were washed with H20, brine, dried over Na2S04 and evaporated under vacuum to give 1 -(4-hydroxy-phenyl)-cyclopropanecarboxylic acid methyl ester (8.9 g, 95%). Ή NMR (400 MHz, CDC13) δ 7.20-7.17 (m, 2 H), 6.75-6.72 (m, 2 H), 5.56 (s, 1 H), 3.63 (s, 3 I I), 1 .60- 1.57 (m, 2 H), 1. 17- 1.15 (m, 2 I I).
[00342] l-(4-Hydroxy-3,5-diiodo-phenyl)-cyclopropanecarboxylic acid methyl ester
[00343] To a solution of l -(4-hydroxy-phenyl)-cyclopropanecarboxylic acid methyl ester (8.9 g, 46 mmol) in CH3CN (80 mL) was added NIS ( 15.6 g, 69 mmol). The mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated and the residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 10: 1 ) to give 1 -(4-hydroxy-3,5-diiodo-phcnyl)-cyclopropanecarboxylic acid methyl ester (3.5 g, 18%). ]H NMR (400 MHz, COClj) δ 7.65 (s, 2 H), 5.71 (s, 1 H), 3.63 (s, 3 H), 1.59- 1.56 (111, 2 11), 1.15- 1.12 (111, 2 11).
[00344] l-[3,5-Diiodo-4-(2-methyI-aIIyIoxy)-phenyI]-cyclopropanecarboxyIic acid methyl ester
[00345] A mixture of 1 -(4-hydroxy-3,5-diiodo-phenyl)-cyclopropanecarboxylic acid methyl ester (3.2 g, 7.2 mmol), 3-chJoro-2-melhyl-propene (1.0 g, 1 1 mmol), K2C03 ( 1.2 g, 8.6 mmol), Nal (0.1 g, 0.7 mmol) in acetone (20 niL) was stirred at 20 °C overnight. The solid was filtered off and the filtrate was concentrated under vacuum to give 1 -[3,5-diiodo-4-(2-melhyl-al]yloxy)-phenyJ]-cyclopropane-carboxy]ic acid methyl ester (3.5 g, 97%). Ή NMR (300 MHz, CDC13) δ 7.75 (s, 2 H), 5.26 (s, 1 H), 5.06 (s, 1 H), 4.38 (s, 2 H), 3.65 (s, 3 H), 1.98 (s, 3H), 1.62-1.58 (m, 2 H), 1 .18- 1.15 (m, 2 H).
[00346] 1 -(3,3-Dimethyl-2,3-dihydro-benzofuran-5-yl)-cyclopropanecarboxylic acid methyl ester
[00347] To a solution of l-[ 3,5-diiodo-4-(2-methyl-allyloxy)-phenyl]-cyclopropane-carboxylic acid methyl ester (3.5 g, 7.0 mmol) in toluene (15 niL) was added BuaSnH (2.4 g, 8.4 mmol) and AIBN (0.1 g, 0.7 mmol). The mixture was heated at reflux overnight. rITic reaction mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 20: 1) to give l -(3,3-dimethyl-2,3-dihydro-bcnzofuran-5-yl)-cyclopropanccarboxylic acid methyl ester ( 1.05 g, 62%). *H NMR (400 MHz, CDC13) δ 7.10-7.07 (in, 2 H), 6.71 (d, J = 8 Hz, 1 H), 4.23 (s, 2 H), 3.62 (s, 3 H), 1.58- 1 .54 (m, 2 H), 1.34 (s, 6 II), 1.17- 1.12 (m, 2 I I).
[00348] l-(3,3-Dimethyl-2,3-dihydrobenzofuran-5-yl)cyclopropanecarboxylic acid
[00349] To a solution of 1 -(3,3-dimemyl-2,3-dihydro-benzofuran-5-yl)-cyclopropanecarboxylic acid methyl ester ( 1.0 g, 4.0 mmol) in MeOH (10 mL) was added LiOH (0.40 g, 9.5 mmol). The mixture was stirred at 40 °C overnight. HCl ( 10%) was added slowly to adjust Uie pH to 5. The resulting mixture was extracted with ethyl acetate (10 mL x 3). The extracts were washed with brine and dried over Na2S04. The solvent was removed under vaccum and the crude product was purified by preparative I 1PLC to give l -(3,3-dimelhyl-2,3-dihydrobenzofuran-5-yl)cyclopropanecarboxylic acid (0.37 g, 1 %). Ή NMR (400 MHz, CDCI3) δ 7.1 1 -7.07 (m, 2 11), 6.71 (d, J = 8 Hz, 1 11), 4.23 (s, 2 I I), 1.66- 1.63 (m, 2 H), 1.32 (s, 6 H), 1.26- 1.23 (m, 2 H).
[00350] Example 10: 2-(7-Methoxybenzo[d][l ,3]dioxol-5-yl)acetonitrile. [00351 ] 3 ,4- Di hy droxy-5- methoxy benzoate
[00352] To a solution of 3,4,5-trihydroxy-benzoic acid methyl ester (50 g, 0.27 mol) and Na2IJ4O7 (50 g) in water (1000 mL) was added Me2S04 ( 120 mL) and aqueous NaOH solution (25%, 200 mL) successively at room temperature. The mixture was stirred at room temperature for 6 h before it was cooled to 0 °C. ITie mixture was acidified to pll ~ 2 by adding cone. I I2SO4 and then filtered. 'ITie filtrate was extracted with HtOAc (500 mL x 3). l ne combined organic layers were dried over anhydrous a2S04 and evaporated under reduced pressure to give methyl 3,4-dihydroxy-5-niethoxybenzoatc (15.3 g 47%), which was used in the next step without further purification.
[00353] Methyl 7-methoxybenzo[d][l,3]dioxoIe-5-carbox late [Θ0354] To a solution of methyl 3,4-dihydroxy-5-methoxybenzoate (15.3 g, 0.0780 mol) in acetone (500 mL) was added CH2BrCl (34.4 g, 0.270 mol) and 2C03 (75.0 g, 0.540 mol) at 80 °C The resulting mixture was heated at reflux for 4 h. lhe mixture was cooled to room temperature and solid K2CO3 was filtered oil'. 'Ihe filtrate was concentrated under reduced pressure, and (he residue was dissolved in EtOAc ( 100 111L). 'Ihe organic layer was washed with water, dried over anhydrous Na2S04, and evaporated under reduced pressure to give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 10: 1) to afford methyl 7-methoxybenzo|d ]| l,31dioxole-5-carboxylate ( 12.6 g, 80%). Ή NMR (400 MHz, CDCI3) δ 7.32 (s, I I I), 7.21 (s, 1 H), 6.05 (s, 2 H), 3.93 (s, 3 H), 3.88 (s, 3 H).
[00355] (7-Methoxybenzo[d][l,3]dioxol-S-yl)methano]
[00356] To a solution of methyl 7-methoxybenzo|d][ 1 ,3]dioxole-5-caiboxylate (14 g, 0.040 mol) in THF ( 100 mL) was added Li A1H4 (3.1 g, 0.080 mol) in portions at room temperature. 'Ihe mixture was stirred for 3 h at room temperature. The reaction mixture was cooled to 0 °C and treated with water (3.1 g) and NaOH ( 10%, 3.1 mL) successively. The slurry was filtered off and washed with TUF. The combined filtrates were evaporated under reduced pressure to give (7-methoxy-benzo[d]| l,3)dioxol-5-y])methanol (7.2 g, 52%). Ή NMR (400 MHz, CDC13) δ 6.55 (s, H I), 6.54 (s, I H), 5.96 (s, 2 H), 4.57 (s, 2 H), 3.90 (s, 3 H).
[00357] 6-(Chloromethyl)-4-methoxybenzo[d][l,3]dioxole
[00358] To a solution of SOCl2 (150 mL) was added (7-methoxybenzo[d|[ l,3]dioxol-5-yl)methanol (9.0 g, 54 mmol) in portions at 0 nC. The mixture was stirred for 0.5 h. The excess SOCl2 was evaporated under reduced pressure to give the crude product, which was basified with sat. aq. NaHCC to pH ~ 7. The aqueous phase was extracted with ljtOAc ( 100 mL x 3). 'Ihe combined organic layers were dried over anhydrous Na2S04 and evaporated to give 6-(chloromethyl)-4-methoxybenzo[d]| l ,3]dioxole ( 10 g 94%), which was used in the next step without further purification. Ή NMR (400 MHz, CDC13) δ 6.58 (s, 1 H), 6.57 (s, 1 H), 5.98 (s, 2 H), 4.51 (s, 2 1-1), 3.90 (s, 3 H).
[00359] 2-(7-Methoxybenzo[d][1 ,3]dioxol-5-y])acetonitri]e
[00360] To a solution of 6-(chloromethyl)-4-methoxybenzo[d |[ 1 ,3]dioxolc (10 g, 40 nimol) in DMSO (100 niL) was added NaCN (2.4 g, 50 mi ol) at room temperature. The mixture was stirred for 3 h and poured into water (500 mL). The aqueous phase was extracted with EtOAc ( 100 mL x 3). The combined organic layers were dried over anhydrous Na2SC>4 and evaporated to give the crude product, which was washed with ether to afford 2-(7-mewoxybenzo[dl| l ,3 |dioxol-5-yl)acetonitrile (4.6 g, 45%). Ή NMR (400 MHz, CDC13) 6 6.49 (s, 2 H), 5.98 (s, 2 H), 3.91 (s, 3 H), 3.65 (s, 2 H). 13C NMR (400 MHz, CDC13) δ 148.9, 143.4, 134.6, 123.4, 1 17.3, 107.2, 101.8, 101.3, 56.3, 23.1.
[00361] Example 1.1 : 2-(3-(Benzyloxy)-4-methoxyphenyl)acetomtrile.
[00362] To a suspension of t-BuO (20.2 g, 0.165 mol) in THF (250 mL) was added a solution of TosMIC ( 16.1 g, 82.6 mmol) in THF ( 100 mL) at -78 °C. The mixture was stirred for 15 minutes, treated with a solution of 3-benzyloxy-4-methoxy-benzaldehyde ( 10.0 g, 51.9 mmol) in THF (50 mL) dropwise, and continued to stir for 1.5 hours at -78 °C. To the cooled reaction mixture was added methanol (50 mL). The mixture was heated at reflux for 30 minutes. Solvent was removed to give a crude product, which was dissolved in water (300 mL). The aqueous phase was extracted with EtOAc (100 mL x 3). The combined organic layers were dried and evaporated under reduced pressure to give crude product, which was purified by column chromatography (petroleum ether/ethyl acetate 10: 1) to afford 2-{3-(bcnzyloxy)-4-mcthoxyphcnyl)- acetonitrile (5.0 g, 48%). Ή NMR (300 MHz, CDC13) δ 7.48-7.33 (m, 5 H), 6.89-6.86 (111, 3 H), 5.17 (s, 2 H), 3.90 (s, 3 H), 3.66 (s, 2 H). I3C NMR (75 MHz, CDC13) 8 149.6, 148.6, 136.8, 128.8, 128.8, 128.2, 127.5, 127.5, 122.1 , 120.9, 1 18.2, 113.8, 1 12.2, 71.2, 56.2, 23.3.
[00363] Example 12: 2-(3-(Benzyloxy)-4-chlorophenyl)acetonitrile.
[00364] (4- C hlo ro-3-h dro xy- phenyl )acetoiiitri le
[00365] BBr3 ( 17 g, 66 mmol) was slowly added lo a solution of 2-(4-chloro-3-methoxyphenyl)acetonitrile ( 12 g, 66 mmol) in dichloromethane (120 mL) al -78 °C under N∑. The reaction temperature was slowly increased lo room temperature. The reaction mixture was stirred overnight and then poured into ice and water. The organic layer was separated, and the aqueous layer was extracted with dichloromethane (40 mL x 3). The combined organic layers were washed with water, brine, dried over Na2S04, and concentrated under vacuum to give (4-chloro-3-hydroxy-phenyl)-acetonitri!e (9.3 g, 85%). Ή NMR (300 MHz, CDC13) δ 7.34 (d, J = 8.4 Hz, 1 H), 7.02 (d, J = 2.1 Hz, 1 H), 6.87 (dd, J = 2. 1 , 8.4 Hz, 1 H), 5.15 (brs, 111), 3.72 (s, 2 H).
[00366] 2-(3-(BenzylQxy)-4-chlorophenyl)acetonitrile
[00367] To a solution of (4-chloro-3-hydroxy-phenyl)acetonitrile (6.2 g, 37 mmol) in CH3CN (80 mL) was added 2C03 ( 10 g, 74 mmol) and BnBr (7.6 g, 44 mmol). The mixture was stirred at room temperature overnight. The solids were filtered off and the filtrate was evaporated under vacuum. 'ITie residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 50: 1) to give 2-(3-(benzyloxy)-4-chlorophenyl)-acetonitrile (5.6 g, 60%). Ή NMR (400 MHz, CDC13) δ 7.48-7.32 (m, 6 H), 6.94 (d, J = 2 Hz, 2 H), 6.86 (dd, J = 2.0, 8.4 Hz, 1 H), 5.18 (s, 2 II), 3.71 (s, 2 H).
[00368] Example 13: 2-(3-(Benzyloxy)-4-methoxyphcnyl)acetonitrilc.
[00369] To a suspension of t-BuO (20.2 g, 0.165 mol) in THF (250 mL) was added a solution of TosMIC (16.1 g, 82.6 mmol) in THF ( 100 mL) at -78 "C The mixture was stirred for 15 minutes, treated with a solution of 3-benzyloxy-4-mclhoxy-benzaldehyde ( 10.0 g, 51.9 mmol ) in THF (50 mL) dropwise, and continued to stir for 1.5 hours at -78 °C To the cooled reaction mixture was added methanol (50 mL). The mixture was heated at reflux for 30 minutes. Solvent of the reaction mixture was removed to give a crude product, which was dissolved in water (300 mL). The aqueous phase was extracted with EtOAc (100 mL x 3). The combined organic layers were dried and evaporated under reduced pressure to give crude product, which was purified by column chromatography (petroleum ether/ethyl acetate 10: 1 ) to afford 2-(3-(benzyloxy)-4-methoxyphenyl)acetonitril (5.0 g, 48%). ]H NMR (300 MHz, CDC13) δ 7.48-7.33 (m, 5 H), 6.89-6.86 (m, 3 II), 5.17 (s, 2 H), 3.90 (s, 3 H), 3.66 (s, 2 H). "C MR (75 MHz, CDCI3) δ 149.6, 148.6, 136.8, 128.8, 128.8, 128.2, 127.5, 127.5, 122.1 , 120.9, 118.2, 1 13.8, 1 12.2, 71.2, 56.2, 23.3.
[00370] Example 1 : 2-(3-Chloro-4-methoxyphenyl)acetonitrile.
[00371] To a suspension of t-BuOK (4.8 g, 40 mmol) in THF (30 mL) was added a solution of TosMIC (3.9 g, 20 mmol) in THF ( 10 mL) at -78 °C. The mixture was stirred for 10 minutes, treated with a solution of 3-chloro-4-methoxy-benzaldehyde (1.7 g, 10 mmol ) in THF (10 mL) dropwise, and continued to stir for 1.5 hours at -78 °C. To the cooled reaction mixture was added methanol ( 10 mL). The mixture was heated at reflux for 30 minutes. Solvent of the reaction mixture was removed to give a crude product, which was dissolved in water (20 mL). The aqueous phase was extracted with EtOAc (20 mL x 3). The combined organic layers were dried and evaporated under reduced pressure to give crude product, which was purified by column chromatography (petroleum ether/ethyl acetate 10: 1) to afford 2-(3-chloro-4-methoxyphenyl)acetonitrile ( 1.5 g, 83%). Ή NMR (400 MHz, CDC13) δ 7.33 (d, J = 2.4 Hz, 1 H), 7.20 (dd, J = 2.4, 8.4 Hz, 1 H), 6.92 (d, J = 8.4 Hz, 1 I -I), 3.91 (s, 3 H), 3.68 (s, 2 H). nC NMR (100 MHz, CDC13) δ 154.8, 129.8, 127.3, 123.0, 122.7, 1 17.60, 1 12.4, 56.2, 22.4.
[00372] Example IS; 2-(3-Fluoro-4-methoxyphenyl)acetonitrile.
[00373] To a suspension of t-BuOK (25.3 g, 0.207 mol) in THF ( 150 mL) was added a solution of' TosMlC (20.3 g, 0.104 mol) in THF (50 mL) at -78 °C. The mixture was stirred for 15 minutes, treated with a solution of 3-fluoro-4-methoxy-benzaldehyde (8.00 g, 51.9 mmol) in THF (50 ml.,) dropwise, and continued to stir for 1.5 hours at -78 °C. To the cooled reaction mixture was added methanol (50 mL). The mixture was heated at reflux for 30 minutes. Solvent of the reaction mixture was removed to give a crude product, which was dissolved in water (200 mL). The aqueous phase was extracted with ElOAc ( 100 mL x 3). The combined organic layers were dried and evaporated under reduced pressure lo give crude product, which was puriRcd by column chromatography (petroleum ether/ethyl acetate 10: 1 ) to afford 2-(3-nuoro-4-methoxyphenyl)acetonilrile (5.0 g, 58%). 1 H NM (400 MHz, CDCh) 8 7.02-7.05 (m, 2 H), 6.94 (t, J = 8.4 Hz, 1 H), 3.88 (s, 3 H), 3.67 (s, 2 H). 13C NMR (100 MHz, CD(¾) 6 152.3, 147,5, 123.7, 122.5, 1 17.7, 1 15.8, 1 13.8, 56.3, 22.6.
[00374] Example 16: 2-(4-Chloro-3-methoxyphenyl)acetonitrile.
Y **1 Mel, eC¾ γγΟ β K^ a CHaCN ^ ^a
[00375] Chloro-2-methoxy-4-methyl-benzene
[00376] To a solution of 2-chloro-5-methyl-phenol (93 g, 0.65 mol) in CH3CN (700 mL) was added CH3I (110 g, 0.78 mol) and 2C03 (180 g, 1.3 mol). The mixture was stirred at 25 °C overnight. The solid was filtered off and the filtrate was evaporated under vacuum to give 1 -chloro-2-mcthoxy-4-methyl-benzene (90 g, 89%). ]H NMR (300 MHz, CDC13) δ 7.22 (d, J = 7.8 Hz, 1 H), 6.74-6.69 (m, 2 H), 3.88 (s, 3 H), 2.33 (s, 3 H).
[00377] 4-Bromomethyl-l-chloro-2-methoxy-benzene
[00378] To a solution of 1 -chJoro-2-methoxy-4-methy]-benzene (50 g, 0.32 mol) in CC14 (350 mL) was added NBS (57 g, 0.32 mol) and ΛΙΒΝ (10 g, 60 mmol). rrhe mixture was heated at reflux for 3 hours, The solvent was evaporated under vacuum and the residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 20: 1) to give 4-bromomelhyl-l-chloro-2-methoxy-benzene (69 g, 92%). Ή NMR (400 MHz, CDC13) δ 7.33-7.31 (ni, 1 I I), 6.95-6. 1 (m, 2 H), 4.46 (s, 2 H), 3.92 (s, 3 H).
[00379] 2-(4-Chloro-3-methoxyp enyl)acetonitrile
[00380] To a solution of 4-bromomethyl-l -chloro-2-melhoxy-ben/.ene (68.5 g, 0.290 mol) in C2H5OH (90%, 500 mL) was added NaCN (28.5 g, 0.580 mol). The mixture was stirred at 60 °C overnight, lilhanol was evaporated and the residue was dissolved in H20. The mixture was extracted with ethyl acetate (300 mL x 3). The combined organic layers were washed with brine, dried over Na2S04 and purified by column chromatography on silica gel (petroleum ether/ethyl acetate 30: 1) to give 2-(4-chloro-3-mcthoxyphcnyl)acctonitrilc (25 g, 48%). Ή NMR (400 MHz, CDC13) δ 7.36 (d, J = 8 Hz, 1 H), 6.88-6.84 (m, 2 H), 3.92 (s, 3 H), 3.74 (s, 2 H). 13C NMR ( 100 MHzt CDCl3) 5 155.4, 130.8, 129.7, 122.4, 120.7, 1 17.5, 1 11.5, 56.2, 23.5.
[00381] Example 17: l-(3-(Hydroxymethyl)-4-methoxyphenyl)cyclopropanecarboxylic acid.
[00382] l-(4-Methoxy-pheny!)-cyclopropanecarboxylic acid methyl ester
[00383] To a solution of f -(4-methoxy-phenyl)-cyclopropanecarboxylic acid (50 g, 0.26 mol) in MeOIl (500 mL) was added loluene-4-sulfonic acid iiionohydrate (2.5 g, 13 mmol) at room temperature. The reaction mixture was heated at reflux for 20 hours. MeOH was removed by evaporation under vacuum and EtOAc (200 mL) was added. The organic layer was washed with sat. aq. NaHCOs ( 100 mL) and brine, dried over anhydrous a2S04 and evaporated under vacuum to give l -(4-methoxy-phenyl)-cyclopropanecarboxylic acid methyl ester (53 g, 99%). Ή NMR (CDC13.400 MHz) 6 7.25-7.27 (m, 2 H), 6.85 (d, / = 8.8 Hz, 2 H), 3.80 (s, 3 H), 3.62 (s, 3 H), 1 .58 (m, 2 H), 1. 15 (in, 2 H).
[00384] l-(3-Chloromethyl-4-methoxy-phenyl)-cyclopropanecarbox lic acid methyl ester
[00385] To a solution of 1 -(4-niethoxy-phenyl)-cyclopropanecarboxylic acid methyl ester (30.0 g, 146 mmol) and MOMCl (29.1 g, 364 mmol) in CS2 (300 mL) was added TiCU (8.30 g, 43.5 mmol) at 5 °C. The reaction mixture was heated at 30 °C for 1 d and poured into ice-water. The mixture was extracted with CH2G2 ( 150 mL x 3). The combined organic extracts were evaporated under vacuum to give l -(3-chloromethyl-4-mcthoxy-phenyl)-cyclopropanecarboxylic acid methyl ester (38.0 g), which was used in the next step without further purification.
[00386] l-(3-Hydroxymethyl-4-methoxy-phenyl)-cyclopropanecarboxylic acid ester
[00387] To a suspension of l-(3-chloromcthyl-4-mcthoxy-phenyl)-cyclopropanccarboxylic acid methyl ester (20 g) in water (350 mL) was added Bu4NBr (4.0 g) and Na2C03 (90 g, 0.85 mol) at room temperature. The reaction mixture was heated at 65 °C overnight. rlTie resulting solution was acidified with aq. HC1 (2 niol/L) and extracted with EtOAc (200 mL x 3). The organic layer was washed with brine, dried over anhydrous N 2S04 and evaporated under vacuum to give crude product, which was purified by column (petroleum ether/ethyl acetate 15: 1) to give l -(3-hydroxymethyl-4-methoxy-phenyl)-cyclopropanecarboxylic acid methyl ester (8.0 g, 39%). !H NMR (CDC13, 400 MHz) 6 7.23-7.26 (m, 2 H), 6.83 (d, J = 8.0 Hz, 1 H), 4.67 (s, 2 H), 3.86 (s, 3 H), 3.62 (s, 3 II), 1.58 (q, J = 3.6 Hz, 2 H), 1.14- 1.17 (m, 2 H).
[00388] l-[3-(iert-Butyl-dimethyl-silanyloxyinethyl)-4-inethoxy-phenyl]cycl()propane carboxylic acid methyl ester
[00389] To a solution of 1 -(3-hydroxymemyl-4-methoxy-pheny])-cyclopropanecarboxylic acid methyl ester (8.0 g, 34 nimol) in CH2CI2 (100 niL) were added imidazole (5.8 g, 85 mmol) and TBSCI (7.6 g, 51 mniol) at room temperature. The mixture was stirred overnight al room temperature. The mixture was washed with brine, dried over anhydrous Na2SC and evaporated under vacuum lo give crude product, which was purified by column (petroleum ether/ethyl acetate 30: 1 ) to give l -[3-(ier/-butyl-dimethyl-silanyloxymethyl)-4-methoxy-phenylj-cyclopropanecarboxylic acid methyl ester (6.7 g, 56%). lH NMR (CDC13, 400 MHz) 6 7.44-7.45 (m, I H), 7.19 (dd, ./ = 2.0, 8.4 Hz, 1 II), 6.76 (d, ./ = 8.4 Hz, 1 H), 4.75 (s, 2 H), 3.81 (s, 3 H), 3.62 (s, 3 H), 1.57- 1.60 (m, 2 H), 1 . 15- 1. 18 (m, 2 H), 0.96 (s, 9 H), 0.1 1 (s, 6 I I).
[00390] l-(3-Hydroxymethyl-4-methoxy-phenyl)-cyclopropanecarboxylic acid
[00391] To a solution of 1 -|3-(/eri-butyl-dimethyl-silanyloxymethyl)-4-methoxy-phenyl]-cyclopropane carboxylic acid methyl ester (6.2 g, 18 mmol) in eOH (75 mL) was added a solution of UOH.H2O (1.5 g, 36 mmol) in water (10 niL) at 0 °C. The reaction mixture was stirred overnight at 40 °C. MeOH was removed by evaporation under vacuum. AcOII (1 mol/1 40 ml.) and RtOAc (200 ml.,) were added. The organic layer was separated, washed with brine, dried over anhydrous N 2S04 and evaporated under vacuum to provide l -(3-hydroxymethy]-4-mcthoxy-phenyl)-cyclopropanccarboxylic acid (5.3 g).
[00392] Example 18: 2-(7-Chlorobenzo[d][l,3]dioxol-S-yl)acetonitrile.
[00393] 3-Chloro-4,5-dihydroxybenzaldehyde
[00394] To a suspension of 3-chloro-4-hydroxy-5-methoxy-benzaldehyde ( 10 g, 54 mmol) in dichloromethane (300 mL) was added BBr3 (26.7 g, 107 mmol) dropwise at -40 °C under N2. After addition, the mixture was stirred at this temperature for 5 h and then was poured into ice water. 'lTie precipitated solid was filtered and washed with petroleum ether. The filtrate was evaporated under reduced pressure to afford 3-chloro-4,5-dihydroxybcnza]dehyde (9.8 g, 89%), which was directly used in the next step.
[00395] 7-Chlorobenzo|d|| l,3]dioxolc-5-carbaldehyde
[00396] To a solution of 3-chloro-4,5-dihydroxybenzaldehyde (8.0 g, 46 mmol) and BrClCH2 (23.9 g, 185 mmol) in dry DMF (100 mL) was added Cs2C03 (25 g, 190 mmol). The mixture was stirred at 60 "C overnight and was then poured into water. The resulting mixture was extracted with EtOAc (50 mL x 3). The combined extracts were washed with brine ( 100 mL), dried over Na2SC>4 and concentrated under reduced pressure to afford 7-chlorobenzo[d][l ,3]dioxole-5-carbaldehyde (6.0 g, 70%). Ή NMR (400 MHz, CDC13) δ 9.74 (s, 1 II), 7.42 (d, 7 = 0.4 Hz, 1 H), 7.26 (d, J = 3.6 Hz, 1 H), 6.15 (s, 2 H).
[00397] (7-Chlorobenzo[d][l,3]dioxol-5-yl)methanol
[00398] To a solution of 7-chlorobenzo[d][ 1 ,3]dioxole-5-carbaldehyde (6.0 g, 33 mmol) in THF (50 niL) was added NaBH, (2.5 g, 64 mmol) ) in portions at 0 aC. The mixture was stirred at this temperature for 30 min and then poured into aqueous NH4CI solution. The organic layer was separated, and the aqueous phase was extracted with EtOAc (50 mL x 3). The combined extracts were dried over Na2S04 and evaporated under reduced pressure to afford (7-chlorobenzo[d ][ l,3]dioxol-5-yl)mcthanol, which was directly used in the next step.
[00399] 4-Chloro-6-(chloromethyl)benzo[d][l,3]dioxole
[00400] Λ mixture of (7-chlorobenzold][ l ,3]-dioxol-5-yl)methanol (5.5 g, 30 mmol) and SOCI2 (5.0 niL, 67 mmol) in dichloromethane (20 niL) was stirred at room temperature for 1 h and was then poured into ice water. The organic layer was separated and the aqueous phase was extracted with dichloromethane (50 mL x 3). Ihe combined extracts were washed with water and aqueous NaHC03 solution, dried over N 2S0 and evaporated under reduced pressure to afford 4-chloro-6-(chloromethyl)benzo|dl| l,3]dioxole, which was directly used in the next step.
[00401] 2-(7-Chlorobenzo[d][l,3]dioxol-5-yl)acetonitrile
[00402] A mixture of 4-chloro-6-(chloromethyl)benzo[d][ l ,3]dioxole (6.0 g, 29 mmol) and NaCN ( 1.6 g, 32 mmol) in DMSO (20 ml.) was stirred at 40 °C for I h and was then poured into water. The mixture was extracted with EtOAc (30 mL x 3). The combined organic layers were washed with water and brine, dried over Na2S04 and evaporated under reduced pressure to afford 2-(7-chlorobenzo[d][ l,3]dioxol-5-yl)acetonitrile (3.4 g, 58%). 1 II N 6 6.81 (s, 1 H), 6.71 (s, 1 I I), 6.07 (s, 2 H), 3.64 (s, 2 H). 13 C-NMR 5149.2, 144.3, 124.4, 122.0, 1 17.4, 114.3, 107.0, 102.3, 23.1.
[00403] Example 19: l-(Benzo[d]oxazol-5-yl)cyclopropanecarboxylic acid.
[00404] l-Benzooxazol-5-yl-cyclopropanecarboxylic acid methyl ester
[00405] To a solution of 1 -(3-amino-4-hydroxypheny])cyclopropanecarboxylic acid methyl ester (3.00 g, 14.5 mmol) in DMF were added triinethyl orthofomiale (5.30 g, 14.5 mmol) and a catalytic amount of p-lolueneslufonic acid monohydrate (0.3 g) at room temperature. The mixture was stirred for 3 hours at room temperature. The mixture was diluted with water and extracted with EtOAc ( 100 mL x 3). The combined organic layers were dried over anhydrous a2SO.j and evaporated under vacuum to give l-benzooxazol-5-yl-cyclopropanecarboxylic acid methyl ester (3.1 g), which was directly used in the next step. Ή NMR (CDC-3.400 MHz) 6 8.09 (s, 1 ), 7.75 (d, J = 1.2 Hz, 1 11), 7.53-7.51 (m, 1 H), 7.42-7.40 (m, 1 H), 3.66 (s, 3 H), 1.69- 1 .67 (m, 2 H), 1.27- 1.24 (m, 2 11).
[00406] l-(Benzo[d]oxazol-5-y])cyclopropanecar oxyIic acid
[00407] To a solution of l-benzooxazol-5-yl-cyclopropanecarboxylic acid methyl ester (2.9 g) in I3tSH (30 mL) was added A1C13 (5.3 g, 40 mmol) in portions at 0 °C. The reaction mixture was stirred for 18 hours at room temperature. Water (20 mL) was added dropwise at 0 °C The resulting mixture was extracted with BlOAc (100 mL x 3). 'Ihe combined organic layers were dried over anhydrous Na2S04 and evaporated under vacuum to give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 1 :2) to give l -(benzold]oxazol-5-yl)cyclopropanecarboxylic acid (280 mg, 1 1 % over two steps). Ή NMR (DMSO, 400 MHz) δ 12.25 (brs, 1 H), 8.71 (s, 1 H), 7.70-7.64 (m, 2 H), 7.40 (dd, J = 1.6, 8.4 Hz, 1 H), 1.49- 1.46 (in, 2 H), 1.21- 1.18 (m, 2 H). MS (ESI) m e (M+H+) 204.4.
[00408] Example 20: 2-(7-Fluorobenzo[d][l,3]dioxol-5-yl)acetonitrile
[00409] 3-FIuoro-4,5-dihydroxy-benzaldehyde
[00410] To a suspension of 3-fluoro-4-hydroxy-5-melhoxy-benzaldehyde (1.35 g, 7.94 rnmol) in dichloromethane ( 100 mL) was added BBr3 ( 1.5 mL, 16 mmol) dropwise at - 78 °C under N2 . Alter addition, the mixture was warmed to - 30 °C and it was stirred at this temperature for 5 h. I "he reaction mixture was poured into ice water. 1 Tie precipitated solid was collected by filtration and washed with dichloromethane to afford 3-fiuoro-4,5-dihydroxy-benzaldehyde ( 1 .1 g, 89%), which was direcdy used in the next step. [00411 ] 7-Fluoro-benzo[1 ,3]dioxole-5-carbaldehyde
[00412] To a solution of 3-fluoro-4,5-dihydroxy-benzaldehyde (1 .5 g, 9.6 mmol) and BrClCH2 (4.9 g, 38.5 mmol) in dry D F (50 mL) was added Cs2C03 (12.6 g, 39 mmol). The mixture was slirred at 60 °C overnighi and was then poured into water. ΊΊιβ resulting mixture was extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine ( 100 mL), dried over Na2S04 and evaporated under reduced pressure to give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetale = 10/1) to afford 7-fluon>bcnzo[ l ,3]dioxole-5-carbaldehyde (0.80 , 49%). 1H NMR (300 MHz, CDCI3) δ 9.78 (d, J = 0.9 Hz, 1 H), 7.26 (dd, J = 1.5, 9.3 Hz, 1 H), 7.19 (d, 7 = 1.2 Hz, 1 H), 6.16 (s, 2 H).
[00413] <7-Fluoro-benzo[l,3]dioxol-5-yl)-methaiiol
[00414] To a solulion of 7-fluoro-benzo[ 1 ,3]dioxole-5-carbaldehyde (0.80 g, 4.7 nimol) in MeOH (50 niL) was added NaBI 14 (0.36 g, 9.4 mniol) in portions at 0 °C. The mixture was stirred at this temperature for 30 min and was then concentrated to dryness. The residue was dissolved in EtOAc. The EtOAc layer was washed with water, dried over 2S04 and concentrated to dryness to afford (7-fluoro-benzo| l ,3]dioxol-5-yl)-melhanol (0.80 g, 98%), which was directly used in the next step.
[00415] 6-Chloromethyl-4-fluoro-benzo[l^]dioxole [004J6] To SOCI2 (20 niL) was added (7-fluoro-benzo| 1 ,3Jdioxol-5-yl)-methano] (0.80 g, 4.7 mrnol) in portions at 0 °C. The mixture was warmed to room temperature over 1 h and then was heated at reflux for 1 h. The excess SOCI2 was evaporated under reduced pressure to give the crude product, which was basified with saturated aqueous NallCOi to pll ~ 7. The aqueous phase was extracted with ElOAc (50 niL x 3). The combined organic layers were dried over Na2S04 and evaporated under reduced pressure to give 6-chloromethyl-4-fluoro-benzo[ 1 ,3]dioxole (0.80 g, 92%), which was directly used in the next step.
[00417] 2-(7-Fluorobenzo[d][l,3]dioxol-5-yl)acetonilriIe
[00418] A mixture of 6-chloromethyl-4-fluoro-benzo| l,3]dioxole (0.80 g, 4.3 mmol) and NaCN (417 mg, 8.51 mmol) in D SO (20 niL) was stirred at 30 °C for 1 h and was then poured into water. The mixture was extracted with EtOAc (50 mL x 3). 'Ihe combined organic layers were washed with water (50 mL) and brine (50 mL), dried over Na2S04 and evaporated under reduced pressure to give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 10/1) to afford 2-(7- fluorobenzold][ l ,3]tli xol-5-yl)acetonitri le (530 mg, 70%). l NMR (300 MHz, CDClj) 6 6.68-6.64 (m, 2 H), 6.05 (s, 2 II), 3.65 (s, 2 H). 1 C-NMK 5151.1, 146.2, 134.1 , 124.2, 1 17.5, 1 10.4, 104.8, 102.8, 23.3.
[00419] Example 21 : l-(lH-Indol-5-yl)cyclopropanecarboxylic acid
[00420] Methyl l-phenylcyclopropanecarboxylate
[00421] To a solution of 1 -phenylcyclopropanecarboxylic acid (25 g, 0.15 mol) in CH3OH (200 mL) was added TsOH (3 g, 0.1 mol) at room temperature. The mixture was refluxed overnight. The solvent was evaporated under reduced pressure to give crude product, which was dissolved into EtOAc. ITie EtOAc layer was washed with aq. sat. NaHC03- The organic layer was dried over anhydrous Na2S04 and evaporated under reduced pressure to give methyl 1-phenylcyclopropanecarboxylate (26 g, 96%), which was used directly in the next step. Ή NMR (400 MHz, CDCb) δ 7.37-7.26 (in, 5 H), 3.63 (s, 3 H), 1.63- 1.60 (m, 2 H), 1.22-1.1 (m, 2 H).
[00422] Methyl l-(4-nitrophenyl)cyclopropanecarboxylate
[00423] To a solution of 1 -phenylcyclopropanecarboxylate (20.62 g, 0.14 mol) in HzSO C^Clz (40 mL/40 mL) was added KN03 (12.8 g, 0.13 mol) in portion at O °C. The mixture was stirred for 0.5 hr at 0 °C. Ice water was added and the mixture was extracted with EtOAc (100 mL x 3). The organic layers were dried with anhydrous NU2SO4 and evaporated to give methyl l -(4-nitrophenyl)cyclopropanecarboxylate (21 g, 68%), which was used directly in the next step. Ή N R (300 MHz, CDC13) δ 8. 18 (dd, J = 2. 1 , 6.9 Hz, 2 H), 7.51 (dd, .7 = 2.1 , 6.9 Hz, 2 I I), 3.64 (s, 3 H), 1.72- 1.69 (in, 2 H), 1 .25- 1.22 (m, 2 H).
[00424] Methyl 1 -(4-aminophenyl)cyclopropanecar oxylate
[00425] To a solution of methyl l -(4-nilrophenyl)cyclopropanecarboxylate (20 g, 0.09 mol) in MeOH (400 mL) was added Ni (2 g) under nitrogen atmosphere. The mixture was stirred under hydrogen atmosphere ( 1 atm) at room temperature overnight. The catalyst was filtered off through a pad of Celite and the filtrate was evaporated under vacuum to give crude product, which was purified by chromatography column on silica gel (petroleum ether/ethyl acetate = 10: 1 ) to give methyl l -(4-aniinophonyl)eyclopropanecarboxylate (1 1 .38 g, 66%). Ί Ι NMR (300 MHz, CDC13) δ 7. 1 6 (d, ./ = 8. 1 Hz, 2 H), 6.86 (d, ./ = 7.8 Hz, 2 H), 4.31 (br, 2 H), 3.61 (s, 3 H), 1.55- 1.50 (111, 2 H), 1.30- 1 .12 (m, 2 H).
[00426] Methyl l -(4-amino-3-bromoplienyl)cyclopropanecarboxylate
[00427] To a solution of methyl l -(4-aminophenyl)cyclopropanecarboxylate ( 10.38 g, 0.05 mol) in acetonitrile (200 mL) was added NBS (9.3 g, 0.05 mol) at room temperature. The mixture was stirred overnight. Water (200 mL) was added. 1¾e organic layer was separated and the aqueous phase was extracted with EtOAc (80 mL x3). The organic layers were dried with anhydrous Na2SC>4 and evaporated to give methyl l -(4-amino-3-bromophenyl)cyclopropanccarboxylate ( 10.6 g, 78%), which was used directly in the next step. Ή NMR (400 MHz, CDCU) δ 7.38 (d, J = 2.0 Hz, 1 H), 7.08 (dd, J = 1.6, 8.4 Hz, I H), 6.70 (d, J = 8.4 Hz, 1 H), 3.62 (s, 3 H), 1 .56- 1 .54 (m, 2 H), 1. 14- 1. 1 l (m, 2 H).
[00428] Methyl 1 -(4-amino-3-((trimethylsiIyl)ethynyl)phenyl)cyclopropane carboxylate
[00429] To a degassed solution oi methyl l-(4-aniino-3-broinophenyl)eyclopropane carboxylate (8 g, 0.03 mol) in Et3N (100 mL) was added ethynyl-tritnetbyJ-silane (30 g, 0.3 mol), DMAP (5% mol) and Pd(PPh3)2Cl2 (5% mol) under N2- The mixture was refluxed at 70 °C overnight. The insoluble solid was filtered off and washed with EtOAc (100 mL x 3). The filtrate was evaporated under reduced pressure to give a residue, which was purified by chromatography column on silica gel (petroleum ether/ethyl acetate =20: 1 ) to give methyl 1-(4-amino-3-((trimethylsilyl)ethynyl)phenyl)cyelopropanecarboxylate (4.8 g, 56%). Ή NMR (300 MHz, CDC13) 67.27 (s, 1 H), 7.10 (dd, J = 2.1, 8.4 Hz, 1 H), 6.64 (d, J - 8.4 Hz, 1 H), 3.60 (s, 3 H), 1.55- 1.51 (m, 2 H), 1.12-1.09 (m, 2 H), 0.24 (s, 9 H).
[00430] Methyl l-(lH-indol-5-yl)cyclopropanecarboxyIate
[00431] To a degassed solution of methyl l-(4-amino-3-((trimethylsilyl)ethynyl)phenyl) cyclopropanecarboxylate (4.69 g, 0.02 mol) in DMF (20 mL) was added Cul (1.5 g, 0.008 mol) under N2 at room temperature. The mixture was stirred for 3 hr at room temperature. The insoluble solid was filtered off and washed with EtOAc (50 mL x 3). The filtrate was evaporated under reduced pressure to give a residue, which was purified by chromatography column on silica gel (petroleum ether/ethyl acetate =20: 1) to give methyl l -(l H-indol-5-yl)cyclopropanecarboxylatc (2.2 g, 51 %). Ή NMR (400 MHz, CDC13) δ 7.61 (s, 1 H), 7.33 (d, 7 = 8.4 Hz, 1 H), 7.23-7.18 (in, 2 H), 6.52-6.51 (m, 1 H) 3.62 (s, 3 H), 1.65- 1.62 (in, 2 II), 1.29-1.23(m, 2 H).
[00432] 1 -(1 H-Indol-5-yl)cyclopropaiiecarboxylic acid
[00433] To a solution of methyl l-(lH-indol-5-yl)cyclopropanecarboxylate (1.74 g, 8 mmol) in CH3OH (50 ra L) and water (20 mL) was added LiOH (1.7 g, 0.04 mol). The mixture was heated at 45 °C for 3 hr. Water was added and the mixture was acidified with concentrated HC1 to pll ~3 before being extracted with EtOAc (20 mL x 3). The organic layers were dried over anhydrous Na2S04 and evaporated to give 1 -(lH-indol-5-yDcyclopropanecarboxylic acid (1.4 g, 87%). Ή NMR (300 MHz, DMSO-f/6) 7.43 (s, 1 H), 7.30-7.26 (m, 2 H), 7.04 (dd, J = 1.5, 8.4 Hz, 1 M), 6.35 (s, 1 H), 1.45- 1.41 (m, 2 H), 1 .14-1. 10 (m, 2 H).
[00434] Example 22: l-(4-Oxochroman-6-yl)cyclopropanecarboxylic acid
[00435] l -[4-(2-iijrt-Butoxycarbonyl-ethoxy)-phenyl]-cyclopropanecarboxylic methyl ester
[00436] To a solution of 1 -(4-hydroxy-pheny!)-cyclopropanecarboxylic methyl ester (7.0 g, 3.6 mmol) in acrylic ½rr-butyl ester (50 mL) was added Na (42 mg, 1.8 mmol) at room temperature. The mixture was heated at 1 10 °C for 1 h. After cooling to room temperature, the resulting mixture was quenched with water and extracted with EtOAc (100 mL x 3). The combined organic extracts were dried over anhydrous Na2S04 and evaporated under vacuum to give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 20: 1 ) to give l -| 4-(2-ier/-butoxycarbonyl-ethoxy)-phenyl]-cyclopropanecarboxylic methyl ester (6.3 g, 54%) and unreacted start material (3.0 g). Ή NMR (300 MHz, CDC13) δ 7.24 (d, J = 8.7 Hz, 2 H), 6.84 (d, J = 8.7 Hz, 2 H), 4.20 (t, J = 6.6 Hz, 2 H), 3.62 (s, 3 H), 2.69 (t, J = 6.6 Hz, 2 1-1), 1 .59- 1.56 (m, 2 H), 1.47 (s, 9 H), 1. 17-1 .42 (m, 2 H).
[00437] l-[4-(2-Carboxy-ethoxy)-phenyl]-cyclopropanecarboxylic methyl ester
[00438] A solution of l -[4-(2-iffi-butoxycarbonyl-ethoxy)-phenyl]-cyclopropanecarbox lic methyl ester (6.3 g, 20 mmol) in HC1 (20%, 200 mL) was heated at 1 10 °C for 1 h. After cooling to room temperature, the resulting mixture was filtered. ITi solid was washed with water and dried under vacuum to give l -[4-(2-carboxy-ethoxy)-phenyl]-cyclopropanecarboxylic methyl ester (5.0 g, 96%). ]H NMR (300 MHz, DMSO) 7.23-7. 1 (in, 2 H), 6.85-6.81 (m, 2 I I), 4.13 (l, J = 6.0 Hz, 2 11), 3.51 (s, 3 I I), 2.66 (t, J = 6.0 Hz, 2 U), 1.43- 1.39 (m, 2 H), 1.14- 1.10 (m, 2 I I).
[00439] l-(4-Oxochroman-6-yl)cyclopropaiiecarboxylic acid
[00440] To a solution of 1 -14-(2-carboxy-ethoxy)-phenyl]-cyclopropanecarboxylic methyl ester (5.0 g, 20 mmol) in CH2CI2 (50 niL) were added oxalyl chloride (4.8 g, 38 mmol) and two drops of DMF at 0 °C. 'ITie mixture was stirred at 0-5 °C for 1 h and then evaporated under vacuum. To the resulting mixture was added CH2CJ2 (50 111L) at 0 °C and stirring was continued at 0~5 °C for 1 h. rHie reaction was slowly quenched with water and was extracted with EtOAc (50 111L x 3). The combined organic extracts were dried over anhydrous a2S0 and evaporated under vacuum to give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 20: 1 -2: 1) to give l-(4-oxochroman-6-yl)cyclopropanecarboxylic acid (830 mg, 19%) and methyl l -(4-oxochiOman-6-yl)cyclopropanecarboxylale (1.8 g, 38%). l -(4-Oxochroman-6-yl)cyclopropane-carboxylic acid: Ή NMR (400 MHz, DMSO) δ 12.33 (br s, 1 H), 7.62 (d, / = 2.0 Hz, 1 IT), 7.50 (dd, J = 2.4, 8.4 Hz, 1 H), 6.95 (d, J = 8.4 Hz, 1 H), 4.50 (t, ./ = 6.4 Hz, 2 H), 2.75 (t, J = 6.4 Hz, 2 H), 1.44- 1.38 (m, 2 H), 1.10- 1.07 (m, 2H). MS (ESI) m/z (M+H+) 23 1.4. l-(4-Oxochroman-6-y cyclopropanecarboxylate: lH NMR (400 MHz, CDC13) δ 7.83 (d, J = 2.4 Hz, I H), 7.48 (dd, J = 2.4, 8.4 Hz, 1 H), 6.93 (d, J = 8.4 Hz, 1 IT), 4.55-4.52 (m, 2 H), 3.62 (s, 3 H), 2.80 (t, ,/ = 6.4 Hz, 2 H), 1.62- 1.56 (m, 2 H), 1.18- 1.15 (m, 2H). [00441 ] Example 23: 1 -(4-Hydroxy-4-methoxychroman-6-yl)cyclopropanecarboxylic acid
[00442] 1 -(4-Hydroxy-4-methoxychroman-6-yl)cyclopropanecarbox lie acid
[00443] To a solution of" methyl 1 -(4-oxochroman-6-yl)cyclopropanecarboxylate ( 1.0 g, 4.1 mmol) in MeOH (20 niL) and water (20 niL) was added LiOH-H20 (0.70 g, 16 mmol ) in portions at room temperature. The mixture was stirred overnight at room temperature before the MeOH was removed by evaporation under vacuum. Water and Et20 were added to the residue and the aqueous layer was separated, acidified with HC1 and extracted with EtOAc (50 niL x 3). The combined organic extracts dried over anhydrous N jSCXs and evaporated under vacuum to give l -(4-hydroxy-4-methoxychrotnan-6-yl)cyclopropanecarboxylic acid (480 mg, 44%). Ή NMR (400 MHz, CDCl3) δ 12. 16 (s, 1 H), 7.73 (d, J = 2.0 Hz, 1 H), 7.47 (dd, J = 2.0, 8.4 Hz, 1 1-1), 6.93 (d, J = 8.8 Hz, 1 H), 3.83-3.80 (m, 2 H), 3.39 (s, 3 H), 3.28-3.25 (in, 2 H), 1.71 - 1.68 (m, 2 H), 1.25- 1.22 (m, 2H). MS (ESI) m/z (M+H+) 263.1.
[00444] Example 24: l-(4-Hydroxy-4-methoxychroman-6-yl)cyclopropanecarboxylic acid
[00445] l-Ctiroman-6-yl-cyclopropanecarboxylic methyl ester
[00446] To trifluoroacetic acid (20 mL) was added NaBI-U (0.70 g, 130 mmol) in portions at 0 °C under N2 atmosphere. After stirring for 5 min, a solution of l-(4-oxo-chroman-6-yl)-cyclopropanecarboxylic methyl ester ( 1.6 g, 6.5 mmol) was added at 15 °C. The reaction mixture was stirred for 1 h at room temperature before being slowly quenched with water. The resulting mixture was extracted with EtOAc (50 mL x 3). The combined organic extracts dried over anhydrous Na2S0 and evaporated under vacuum to give l -chroman-6-yl-cyclopropanecarboxylic methyl ester ( 1.4 g, 92%), which was used directly in the next step. Ή NMR (300 MHz, CDClj) δ 7.07-7.00 (m, 2 H), 6.73 (d, J = 8.4 Hz, 1 H), 4.17 (t, J = 5.1 Hz, 2 H), 3.62 (s, 3 H), 2.79-2.75 (m, 2 H), 2.05-1.96 (m, 2 H), 1.57- 1.54 (111, 2 H), 1.16- 1.13 (m, 2H). 1 -(4-Hydroxy-4-methoxychroman-6-yl)cyc)opropanecarboxylic acid
[00448] To a solution of 1 -chroman-6-yl-cyclopropanecarhoxylic methyl ester ( 1.4 g, 60 mmol) in MeOH (20 mL) and water (20 mL) was added LiOH H20 (1.0 g, 240 mmol ) in portions at room temperature. The mixture was stirred overnight at room temperature before the MeOl I was removed by evaporation under vacuum. Water and Et20 were added and the aqueous layer was separated, acidified with HC1 and extracted with EtOAc (50 mL x 3). The combined organic extracts dried over anhydrous N 2S0 and evaporated under vacuum to give l -(4-Hydroxy-4-methoxychroman-6-yl)cyclopropanecarboxylic acid ( 1.0 g, 76%). Ή NMR (400 MHz, DMSO) 6 12.10 (br s, J H), 6.95 (d, J = 2.4 Hz, 2 H), 6.61 -6.59 (m, 1 11), 4.09-4.06 (m, 2 H), 2.70-2.67 (m, 2 H), 1.88- 1.86 (m, 2 H), 1.37- 1.35 (ra, 2 I l>, 1.04- 1.01 (m, 2H). MS (ESI) m/z (M+H+) 217.4.
[00449] Example 25: l-(3-MethyIbenzo[d]isoxazol-5-yl)cyclopropanecarboxy lie acid
[00450] 1 -(3-Acetyl-4-hydroxy-phenyl)-cyclopropanecarboxylic meth l ester
[00451] To a stirred suspension of AlC (58 g, 440 mmol) in CS2 (500 mL) was added acetyl chloride (7.4 g, 95 mmol) at room temperature. After stirring for 5 min, methyl l -(4-methoxyphenyl)cyclopropanecarboxylate (15 g, 73 mmol) was added. The reaction mixture was heated at reflux for 2 h before ice water was added carefully to the mixture at room temperature. The resulting mixture was extracted with EtOAc ( 150 mL x 3). The combined organic extracts were dried over anhydrous Na2S04 and evaporated under reduced pressure to give l-(3-acetyl-4-hydroxy-phenyl)-cyclopropanecarboxylic methyl ester (15 g, 81 %), which was used in the next step without further purification. Ή NMR (CDC13.400 MHz) δ 12.28 (s, 1 H), 7.67 (d, J = 2.0 Hz, 1 1 1), 7.47 (dd, ./ = 2.0, 8.4 Hz, 1 H), 6.94 (d, ./ = 8.4 Hz, 1 I I), 3.64 (s, 3 H), 2.64 (s, 3 H), 1.65- 1.62 (m, 2 H), 1.18- 1. 16(m, 2 H).
[00452] l-[4-Hydroxy-3-(l-hydroxyimino-ethyl)-pheny]]-cyclopropanecarboxylic methyl ester
[00453] To a stirred solution of l -(3-acetyl-4-hydroxy-phenyl)-cyclopropanecarboxylic methyl ester ( 14.6 g, 58.8 mmol) in EtOH (500 mL) were added hydroxylamine hydrochloride (9.00 g, 129 mmol) and sodium acetate (J 1.6 g, 141 mmol) at room temperature. The resulting mixture was heated at reflux overnight. After removal of EtOH under vacuum, water (200 mL) and EtOAe (200 mL) were added. The organic layer was separated and the aqueous layer was extracted with ElOAc (100 mL x 3). The combined organic layers were dried over anhydrous Na2S0 and evaporated under vacuum to give 1 -|4-hydroxy-3-( l -hydroxyimino-ethyl)-phenyl]-cyelopropanecarboxylic methyl ester (14.5 g, 98%), which was used in the next step without further purification. Ή NMR (CDCI3.400 MHz) δ 1 1 .09 (s, 1 I I), 7.39 (d, ./ = 2.0 Hz, 1 H), 7.23 {d, J = 2.0 Hz, 1 H)( 7.14 (s, 1 II), 6. 1 (d, ./ = 8.4 Hz, 1 H), 3.63 (s, 3 H), 2.36 (s, 3 H), 1.62- 1.59 (m, 2 H), 1.18- 1.15 (m, 2 H).
[00454] (/i)-MethyI l-(3-(l-(acetoxyimino)ethyl)-4-hydroxyphenyl)cyclopropane carboxylate
[00455] 'I'he solution of l -|4-hydroxy-3-( l -hydroxy! mino-ethyl)-phenyl]-cyclopropanecarboxylic methyl ester (10.0 g, 40.1 mmol) in Ac20 (250 mL) was heated at 45 °C for 4 h. The Ac20 was removed by evaporation under vacuum before water ( 100 mL) and EtOAc ( 100 mL) were added. 'Yhe organic layer was separated and the aqueous layer was extracted with EtOAc ( 100 mL x 2). The combined organic layers were dried over anhydrous Na2S04 and evaporated under vacuum to give (£)-methyl l-(3-(i -(acetoxyimino)ethyl)-4-hydroxyphenyl)cyclopropanecarboxylate ( 10.5 g, 99%), which was used in the next step without further purification.
[00456] Methyl l-(3-methylbenzo[d]isoxazol-5-yl)cyclopropanecarboxylate
[00457] A solution of (£ methyl l-(3-( 1 -(acetoxyimino)cthyl)-4-hydroxyphenyl)cyclopropanc carboxylate ( 10.5 g, 39.6 mmol) and pyridine ( 1 .3 g, 396 mmol) in DMF ( 150 mL) was heated at 125 °C for 10 h. The cooled reaction mixture was poured into water (250 mL) and was extracted with ElOAe (100 mL x 3). '["he combined organic layers were dried over anhydrous Na2SC>4 and evaporated under vacuum lo give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 50: 1 ) to give methyl l -(3-methylbenzol d]isoxazoI-5-yl)cyclopropaneearboxylate (7.5 g, 82%). Ή NMR (CDC13 300 MHz) δ 7.58-7.54 (m, 2 H), 7.48 (dd, J = 1 .5, 8.1 Hz, I H), 3.63 (s, 3 H), 2.58 (s, 3 H), 1 .71 - 1 .68 (m, 2 H), 1 .27- 1 .23 (m, 2 H).
[00458] 1 -(3-Methylbenzo[d]isoxazoI-5-yl)cyclopropanecarboxylic acid
[00459] To a solution of methyl l-(3-methylbenzo[d|isoxazol-5-yl)cyclopropanecarboxylaLe (1.5 g, 6.5 mmol) in MeOH (20 mL) and water (2 mL) was added Li()H-H20 (0.80 g, 19 mmol ) in portions at room temperature. The reaction mixture was stirred at room temperature overnight before the MeOH was removed by evaporation under vacuum. Water and 1¾0 were added and the aqueous layer was separated, acidified with HC1 and extracted with HtOAc (50 mL x 3). The combined organic extracts were dried over anhydrous Na2S0 and evaporated under vacuum lo give l-(3-meihylbenzo[dJisoxazoI-5-yl)cyclopropanecarboxylie acid (455 mg, 32%). Ή NMR (400 MHz, DMSO) δ 1 2.40 (br s, 1 H), 7.76 (s, 1 H), 7.60-7.57 (m, 2 H), 2.63 (s, 3 H), 1 .52-1.48 (m, 2 H). 1.23- 1 .19 (m, 2H). MS (LSI) m/z (Μ+1-Γ) 218. 1.
[00460] Example 26: l-(Spiro[benzo[d][l,3]dioxole-2,l '.cyclobutane]-5-yljcyclopropane carboxylic acid
[00461] l-(3,4-Dihydroxy-phenyI)-cyclopropanecarboxylic methyl ester
[00462] To a solution of l -(3,4-dihydroxyphcnyl)cyclopropanecarboxylic acid (4.5 g) in MeOH (30 mL) was added TsOH (0.25 g, 1 .3 mmol). The stirring was continued al 50 °C overnight before the mixture was cooled to room temperature. The mixture was concentrated under vacuum and the residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 3: 1 ) to give l -(3,4-dihydroxy-phenyl)-cyclopropanecarboxylic methyl ester (2. 1 g). Ή NMR (DMSO 300 MHz) δ 8.81 (brs, 2 H), 6.66 (d, ./ = 2. 1 Hz, 1 H), 6.61 (d, J = 8.1 Hz, 1 H), 6.53 (dd, J = 2. 1 , 8.1 Hz, 1 H), 3.51 (s, 3 H), 1.38- 1.35 (m, 2 H), 1 .07- 1.03 (m, 2 H).
[00463] Methyl 1-(spiro[benzo[d][l,3]dioxole-2, -cyclobutane]-5-yl)cyclopropane car boxy late
[00464] To a solution of l -(3,4-dihydroxy-phenyl)-cyclopropanecarboxylic methyl ester (1.0 g, 4.8 mmol) in toluene (30 ml .) was added TsOH (0.10 g, 0.50 mmol) and cyclobutanone (0.70 g, 10 mmol). The reaction mixture was heated at reflux for 2 h before being concentrated under vacuum. The residue was purified by chromatography on silica gel (petroleum ether/ethyl acetate 15: 1 ) to give methyl l -(spiro[benzo[d][ l ,3]dioxole-2, i '-cyclobutane]-5-yl)cyclopropanecarboxylate (0.6 g, 50%). Ή NMR (CDC 300 MHz) δ 6.78-6.65 (m, 3 H), 3.62 (s, 3 H), 2.64-2.58 (m, 4 H), 1.89- 1.78 (m, 2 H), 1.56- 1.54 (m, 2 H), 1.53- 1.12(m, 2 H).
[00465] l-(Spiro[benzotd][],3]dioxole-2,l'-cyclobutane]-5-yl)cyclopropane carboxylic acid
[00466] To a mixture of methyl l -(spiro[bcnzo[d][ l,3]dioxole-2, -cyclobutane]-5-yl)cyel-opropanecarboxylaie (0.60 g, 2.3 mmol) in THF H20 (4: 1 , 10 mL) was added LiOH (0.30 g, 6.9 mmol). 'i'he mixture was stirred at 60 °C for 24 h. HC1 (0.5 N) was added slowly to the mixture at 0 °C until pM 2-3. ine mixture was extracted with EtOAc (10 mL x 3). The combined organic phases were washed with brine, dried over anhydrous MgSO
[00467] Example 27: 2-(2T3-Dihydrobenzo[b][l)4]djoxin-6-y])acetonitrile
[00468] 2,3-Dihydro-benzo[l,4]dioxine-6-carboxylic acid ethyl ester
[00469] To a suspension of Cs2C03 (270 g, 1.49 mol) in DMF (1000 mL) were added 3,4-dihydroxybenzoic acid ethyl ester (54.6 g, 0.3 mol) and 1 ,2-dibromoe thane (54.3 g, 0.29 mol) at room temperature. The resulting mixture was stirred at 80 °C overnight and then poured into ice-water. The mixture was extracted with EtOAc (200 mL x 3). The combined organic layers were washed with water (200 mL x 3) and brine ( 100 mL), dried over Na2SO^ and concentrated to dryness. The residue was purified by column (petroleum ether/ethyl acetate 50: 1) on silica gel to obtain 2,3-dihydro-benzo[ l,4]dioxine-6-carboxylic acid ethyl ester ( 18 g, 29%). T l NM (300 MHz, CDC13) δ 7.53 (dd, J = 1.8, 7.2 Hz, 2 I I), 6.84-6.87 (m, 1 H), 4.22-4.34 (m, 6 H), 1.35 (t, J = 7.2 Hz, 3 H).
[00470] (2,3-Dihydro-benzo[l,4]dioxin-6-yl)-methanol
[00471] To a suspension of L1AIH4 (2.8 g, 74 inmol) in THF (20 mL) was added dropwise a solution of 2,3-dihydro-benzoll,4]dioxine-6-carboxylic acid ethyl ester (15 g, 72 mmol) in Π IF ( 10 niL) at 0 °C under N2. The mixture was stirred at room temperature for 1 h and then quenched carefully with addition of water (2.8 mL) and NaOI I ( 10%, 28 mL) with cooling. The precipitated solid was filtered off and the filtrate was evaporated to dryness to obtain (2,3-dihydro-benzo| 1 ,4]dioxin-6-yl)-methanol (10.6 g). Ή N R (300 MHz, DMSO-d6) S 6.73-6.78 (m, 3 II), 5.02 (t, J = 5.7 Hz, 1 H), 4.34 (d, J = 6.0 Hz, 2 H), 4.17-4.20 (m, 4 H).
[00472] 6-Chloromethyl-2,3-dihydro-benzo[l ,4]dioxine
[00473] Λ mixture of (2,3-dihydro-benzo[ 1 ,4|dioxin-6-yl)melhanol (10.6 g) in SOCl2 (10 mL) was stirred at room temperature for 10 min and then poured into ice-water. The organic layer was separated and the aqueous phase was extracted with dichloromethane (50 mL x 3). The combined organic layers were washed with NaHCOs (sal solution), water and brine, dried over Na2S04 and concentrated to dryness to obtain 6-chloromethyl-2,3-dihydro-benzo| 1 ,4]dioxine (12 g, 88% over two steps), which was used directly in next step.
[00474] 2-(2,3-Dihydrobenzo[b][l,4]dioxin-6-y])acetonitrtle
[00475] A mixture of 6-ch!oromethyl-2,3-dihydro-benzo[ 1 ,4]dioxine (12.5 g, 67.7 mmol) and NaCN (4.30 g, 87.8 mmol) in DMSO (50 mL) was stirred at rt for 1 h. 'ITie mixture was poured into water (150 mL) and then extracted with dichloromethane (50 mL x 4). The combined organic layers were washed with water (50 mL x 2) and brine (50 mL), dried over Na2SC and concentrated to dryness. The residue was purified by column (petroleum ether/ethyl acetate 50: 1 ) on silica gel to obtain 2-(2,3-dihydrobenzo[b][ l,4]dioxin-6-yDacelonitrile as a yellow oil (10.2 g, 86%). Ή-NMR (300 MHz, CD(¾) δ 6.78-6.86 (in, 3 H), 4.25 (s, 4 H), 3.63 (s, 2 H).
[00476] The following Table 2 contains a list of carboxylic acid building blocks that were commercially available, or prepared by one of the three methods described above: Table 2: Carboxylic acid building blocks.
[00477] Specific Procedures: Synthesis of aminoindole building blocks
[00478] Example 28: 3-Methyl-1H-indoI-6-amine
[00479] (3-Nitro-phenyl)-hydrazine hydrochloride salt
[00480] 3-Nitro-phenylamine (27.6 g, 0.2 mol) was dissolved in the mixture of H 0 (40 niL) and 37% HCl (40 mL). A solution of NaN02 (13.8 g, 0.2 mol) in H20 (60 mL) was added to the mixture at 0 °C, and then a solution of SnCl2.H20 (135.5 g, 0.6 mol) in 37% HCl ( 100 mL) was added at that temperature. After stirring at 0 °C for 0.5 h, the insoluble material was isolated by filtration and was washed with water to give (3-nitrophenyOnydrazine hydrochloride (27.6 g, 73%).
[00481] N-(3-Nitro-phenyl)-iV'-propylidene-hydrazine
[00482] Sodium hydroxide solution ( 10%, 15 mL) was added slowly to a stirred suspension of (3-niLrophenyl)hydrazine hydrochloride (1.89 g, 10 mmol) in ethanol (20 mL) until pH 6. Acetic acid (5 mL) was added lo the mixture followed by propionaldehyde (0.7 g, 12 mmol). After stirring for 3 h at room temperature, the mixture was poured into ice-water and the resulting precipitate was isolated by filtration, washed with water and dried in air to obtain (E)- l -(3-nitrophenyl)-2-propylidenehydrazine, which was used directly in the next step.
[00483] 3-Methyl-4-nitro-l H-mdole 3 and 3-methyl-6-nitro-lH- indole
[00484] A mixture of (E)- l-(3-nitrophenyl)-2-propylidenehydrazine dissolved in 85 % Η3ΙΌ4 (20 mL) and toluene (20 mL) was heated at 90-100 °C for 2 h. After cooling, toluene was removed under reduced pressure. The resultant oil was basified to pH 8 with 10 % NaOU. The aqueous layer was extracted with EtOAc (1 0 mL x 3). The combined organic layers were dried, filtered and concentrated under reduced pressure to afford the mixture of 3-niethyl-4-nitro- l H-indole and 3-methyl-6-nitro- lH-indole [ 1.5 g in total, 86 , two steps from (3-nitrophenyl)hydrazine hydrochloride) which was used to the next step without further purification.
[00485] 3-MethyM H-indol-6-amine
[00486] The crude mixture from previous steps (3 g, 17 mmol) and 10% Pd-C (0.5 g) in ethanol (30 mL) was stirred overnight under H2 ( I atm) at room temperature. Pd-C was filtered off and the filtrate was concentrated under reduced pressure. The solid residue was purified by column to give 3-melhyl- l H-indol-6-amine (0.6 g, 24%). ]H NMR (CDCI3) 5 7.59 (br s. 1 H), 7.34 (d, J = 8.0 Hz, 111), 6.77 (s, 1H), 6.64 (s, 1 H), 6.57 (in, 1H), 3.57 (brs, 211), 2.28 (s, 311); MS (ESI) m/e (Μ+Ι-Γ) 147.2.
[00487] Example 29: 3-/ert-Butyl-lH-indol-5-amine
[00488] 3-ftri-Rul\ l-5-nitro- 1 H-indole
[00489] To a mixture of 5-nilro- 1 H-indole (6.0 g, 37 niniol) and A\C- (24 g, 0.18 mol) in CH2C12 ( 100 mL) at 0 °C was added 2-bronio-2-inethyl -propane (8.1 g, 37 iniiiol) dropwise. After being stirred at 15 °C overnight, the mixture was poured into ice (100 mL). The precipitated salts were removed by filtration and the aqueous layer was extracted with CH2CI2 (30 mL x 3). The combined organic layers were washed with water, brine, dried over Na2S04 and concentrated under vacuum to obtain the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 20: 1 ) to give 3-terl-butyl-5-nitro- 1 H-indole (2.5 g, 31 %). ]H NMR (CDCI3, 00 MHz) δ 8.49 (d, J = 1.6 Hz, 1 I I), 8.31 (brs, 1 H), 8.05 (dd, J = 2.0, 8.8 Hz, 1 H), 7.33 (d, ./ = 8.8 Hz, 1 H), 6.42 (d, J = 1.6 Hz, 1 H), 1.42 (s, 9 11).
[00490] 3-½rf-Butyl-1 H-indoI-5-amine
[00491] To a solution of 3-im-butyl-5-nitro- 1 H-indole (2.5 g, 12 mmol) in McOH (30 mL) was added Raney Nickel (0.2 g) under N2 protection. The mixture was stirred under hydrogen atmosphere (1 aim) at 15 nC for 1 h. The catalyst was filtered off and the filtrate was concentrated to dryness under vacuum. The residue was purified by preparative HLPC to afford 3-rerr-butyl- lH-indol-5-amine (0.43 g, 1 %). Ή NMR (CDC13, 400 MHz) δ 7.72 (br.s, I I I), 7.11 (d, J = 8.4 Hz, 1 H), 6.86 (d, J = 2.0 Hz, 1 H), 6.59 (dd, ./ = 2.0, 8.4 Hz, 1 H), 6.09 (d, J = ] .6 Hz, 1 H), 1.37 (s, 9 H); MS (ESI) m e (Μ+ΙΓ) 189.1.
[00492] Example 30: 2-terf-Butyl-6-nuoro-l H-indol-5-amine and 6-tert-buio\y-2-tert-butyl-l H-indol-5-amine
[00493] 2-Bromo-5-fluoro-4-nitroaniline
[00494] To a mixture of 3-fluoro-4-nitroaniline (6.5 g, 42.2 mmol) in AcOH (80 mL) and chloroform (25 mL) was added dropwise Br2 (2.15 mL, 42.2 mmol) at 0 °C. After addition, the resulting mixture was stirred at room temperature for 2 h and then poured into ice water. The mixture was basified with aqueous NaOH ( 10%) to pl l ~ 8.0-9.0 under cooling and then extracted with EtOAc (50 mL x 3). The combined organic layers were washed with water (80 mL x 2) and brine (100 mL), dried over Na2S04 and concentrated under reduced pressure to give 2-bromo-5-fluoro-4-nitroaniline (9 g, 90%). ' I I-NMR (400 MHz, DMSO-f/6) S 8.26 (d, J = 8.0, Hz, H I), 7.07 (brs, 211), 6.62 (d, J = 9.6 Hz, 1H).
[00495] 2-(3,3-DimethyIbul-l-ynyl)-5-fluoro-4-nitroarjiline
[00496] A mixture of 2-bronio-5-lluoro-4-nitroaniline (9.0 g, 38.4 mmol), 3,3-dimethyl-but- 1 -yne (9.95 g, 121 mmol), Cul (0.5 g 2.6 mmol), Pd(PPh3)2(¾ (3.4 g, 4.86 mmol) and Et3N (14 mL, 6.9 mmol) in toluene (100 mL) and water (50 mL) was heated at 70 °C for 4 h The aqueous layer was separated and the organic layer was washed with water (80 mL x 2) and brine (100 mL), dried over Na2S04 and concentrated under reduced pressure to dryness. The residue was rccrystallizcd with ether to afford 2-(3,3-dimcthylbut- l -ynyl)-5-fiuoro-4-nitroaniline (4.2 g, 46%). Ή-NMR (400 MHz, DMSO-rf6) δ 7.84 (d, ./ = 8.4 Hz, 1H), 6.84 (brs, 2H), 6.54 (d, J = 14.4 Hz, IH), 1.29 (s, 9H).
[00497] /V-(2-(3,3-Dimethy]but-l-ynyl)-5-fluoro-4-nitrophenyl)butyramide
[00498] To a solution of 2-(3,3-diraelhylbut- l-ynyl)-5-fluoro-4-nitroatiiIine (4.2 g, 17.8 mmol) in dichloromethane (50 niL) and 1¾N ( 10.3 niL, 71.2 mmol) was added butyryl chloride (1.9 g, 17.8 mmol) at 0 °C. The mixture was stirred at room temperature for 1 h and then poured into water. The aqueous phase was separated and the organic layer was washed with water (50 mL x 2) and brine (100 mL), dried over a2S0 and concentrated under reduced pressure to dryness. The residue was washed with ether to give N-(2-(3,3-diinethylbut- l -ynyl)-5-fluoro-4-nitrophenyl)butyramide (3.5 g, 67%), which was used in the next step without further purification.
[00499] 2-ferf-Butyl-6-fluoro-5-nitro-lH-indo-e
[00500] A solution of N-(2-(3,3-dimethylbul- 1 -ynyl)-5-iluoro~4-nitrophenyl)butyramide (3.0 g, 9.8 mmol) and TBAF (4.5 g, 17.2 mmol) in DMF (25 mL) was heated ut 100 °C overnight. The mixture was poured into water and then extracted with ElOAc (80 mL χ 3). The combined extracts were washed with water (50 mL) and brine (50 mL), dried over Na2S04 and concentrated under reduced pressure to dryness. rrhe residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 20: 1 ) to give compound 2-tcit-bulyl-6-nuoro-5-nitro- l H-indolc (1.5 g, 65%). Ί-1-NMR (400 MHz, CDC13) δ 8.30 (d, J = 7.2 Hz, 1H), 7.12 (d, 7 = 1 1.6 Hz, 1H), 6.35 (d, J = 1.2 Hz, 1 H), 1.40 (s, 911).
[00501] 2-½rr-Butyl-6-fluoro-lH-indol-5-amine
[00502] A suspension of 2-tert-butyl-6-fluoro-5-nitro- lH-indole ( 1.5 g, 6.36 mmol) and Ni (0.5 g) in MeOH (20 mL) was stirred under H2 atmosphere ( 1 atm) at the room temperature for 3 h. The catalyst was filtered off and the filtrate was concentrated under reduced pressure to dryness. The residue was reerystallized in ether to give 2-tert-but.yl-6-riuoro- IH-indol-5-aminc (520 rag, 38%). JH-NMR (300 MHz, D SO-tfc) δ 10.46 (brs, 1H), 6.90 (d, J = 8.7 Hz, I I I), 6.75 (d, 7 - 9.0 Hz, HI), 5.86 (s, 111), 4.37 (brs, 2H), 1.29 (s, 9H); MS (ESI) m/e 206.6.
[00503] 6-iert-Butoxy-2-tert-butyl-5-nitro-lH-indole
[00504] A solution of N-(2-(3,3-dimcthylbut- 1 -ynyl)-5-fluoro-4-nilrophcnyl)butyramide (500 mg, 1.63 mmol) and i-BuO (0.37 g, 3.26 mmol) in D F ( 10 mL) was heated at 70 "C for 2 h. The mixture was poured into water and (hen extracted with EtOAc (50 mL x 3). The combined extracts were washed with water (50 mL) and brine (50 mL), dried over N 2SC>4 and concentrated under reduced pressure to give 6-teit-buloxy-2-lerl-butyl-5-nitro- l H-indolc ( 100 mg , 21 ). Ή-NMR (300 MHz, D SO-rf6) 5 11 .35 (brs, 1 I I), 7.99 (s, 1 I I), 7.08 (s, I H), 6.25 (s, 1 H), 1 .34 (s, 9H), 1 .30 (s, 9H).
[00505] 6-/trrf-Butoxy-2-tert-butyl-1 H-indol-5-amine
[00506] A suspension of 6-ierf-butoxy-2-tert-buty 1-5-nitro- 1 H-indole ( 100 mg, 0.36 mmol) and Raney Ni (0.5 g) in MeOH (15 mL) was stirred under H2 atmosphere (1 atm) at the room temperature for 2.5 h. The catalyst was filtered off and the filtrate was concentrated under reduced pressure to dryness. The residue was reerystallized in ether to give 6-/erf-buloxy-2-/m-butyl-l H-indol-5-aminc (30 mg, 32%). Ή-NMR (300 MHz, McOD) 6.98 (s, I I I), 6.90 (s, 1H), 5.94 (d, ,/ = 0.6 Hz, IH), 1.42 (s, 9H), 1.36 (s, 9H); MS (ESI) m/e 205.0.
[00507] Example 31 : l -it'rf-Butyl-l H-indol-S-amine
[00508] V-ier/-Butyl-4-nitroaniline
[00509] A solution of l -fluoro-4-nitro-benzene ( 1 g, 7.1 mmol) and tert-butylamino ( 1.5 g, 21 mmol) in DMSO (5 niL) was stirred at 75 °C overnight. The mixture was poured into water (10 niL) and extracted with HtOAc (7 mL x 3). The combined organic layers were washed with water, brine, dried over Na2SOa and concentrated under vacuum to dryness. The residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 30: 1 ) to afford N½ri-butyl-4-niLroaniline (1 g, 73%). Ή NMR (CDC13, 400 MHz) δ 8.03-8.00 (m, 211), 6.61 -6.57 (m, 211), 4.67 (brs, II I), 1.42 (s, 91 ]).
[00510] (2- Bronio-4- nil ro-phen) l)-te rt -butyl -amine
[00511] To a solution of Nieri-butyl-4-nitroaniline (1 g, 5.1 mmol) in AcOH (5 mL) was added Br2 (0.86 g, 54 mmol) dropwisc at 15 °C. After addition, the mixture was stirred at 30 "C for 30 min and then filtered. The filter cake was basi fied to pl l 8-9 with aqueous Nal lCOi. The aqueous layer was extracted with EtOAc (10 mL x 3). The combined organic layers were washed with water, brine, dried over Na2S04 and concentrated under vacuum to give (2-bromo-4-nitro-phenyl)-len-buly]-amine (0.6 g, 43%). *H-NMR (CDC13, 400 MHz) δ 8.37 (dd, J = 2.4 Hz, 1H), 8.07 (dd, J = 2.4, 9.2 Hz, 1H), 6.86 (d, / = 9.2 Hz, 1H), 5. 19 (brs, I H), 1.48 (s, 9H).
[00512] iert-Butyl-(4-nitro-2-lrimethylsi.any]ethynyl-phenyl)-amine
[00513] To a solution of (2-bromo-4-nitro-phenyl)-tert-butyl-amine (0.6.g, 2.2 mmol) in Et3N ( 10 mL) was added Pd(PPh3)2Cl2 (70 mg, 0.1 mmol), Cul (20.9 nig, 0.1 mmol) and ethynyl-trimethyl-silane (0.32 g, 3.3 mmol) successively under N2 protection. The reaction mixture was heated at 70 "C overnight. The solvent was removed under vacuum and the residue was washed with EtOAc ( 10 mL x 3). 'ITie combined organic layers were washed with water, brine, dried over Na2S04 and concentrated under vacuum to dryness. The residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 20: 1 ) to afford tert-butyl-(4-nitro-2-lrimethylsiIanylethynyl-phenyl)-amine (100 mg, 16%). JH-NMR (CDC13, 400 MHz) δ 8.20 (d, J = 2.4, Hz, 111), 8.04 (dd, ./ = 2.4, 9.2 Hz, 1 H), 6.79 (d, J = 9.6 Hz, 1 H), 5.62 (bra, 111), 1.41 (s, 9H), 0.28 (s, 9H).
[00514] 1 -tert-Butyl-5-nitro-l H-indole
[00515] To a solution of tert-butyl-(4-nitro-2-triinethylsilanylethynyl-phenyl)-aniine ( 10 mg, 0.035 mmol) in DM17 (2 mL), was added Cul (13 mg, 0.07 mmol) under N2 protection. The reaclion mixture was stirred at 100 °C overnight. At this time, EtOAc (4 mL) was added to the mixture. The mixture was filtered and the filtrate was washed wilh water, brine, dried over Na2S04 and concentrated under vacuum to obtain l-tert-butyl-5-nitro- l H-indole (7 mg, 93%). 'I I-NMR (CDCI3, 300 MHz) δ 8.57 (d, J = 2.1 Hz, I H), 8.06 (dd, J = 2.4, 9.3 Hz, 1 H), 7.65 (d, 7 = 9.3 Hz, 1 H), 7.43 (d, 7 = 3.3 Hz, 1 H), 6.63 (d, .7 = 3.3 Hz, 1H), 1.76 (s, 9H).
[00516] 1 -torf-Butyl-lH-indol-5-amine
[00517] To a solution of 1 -wrt-huiyl-5-riiiro-l H-indole (6.5 g, 0.030 mol) in MeOH (100 mL) was added Raney Nickel (0.65 g, 10%) under N2 protection. The mixture was stirred under hydrogen atmosphere ( 1 aim) at 30 UC for 1 h. The catalyst was filtered off and the filtrate was concentrated under vacuum to dryness. The residue was purified by column chromatography on silica gel (Pli EtOAc 1 :2) to give l -zm-butyl- lH-indol-5-amine (2.5 g, 45%). 1H-NMR (CDC13, 400 MHz) δ 7.44 (d, J = 8.8 Hz, 1H), 7.19 (dd, J = 3.2 Hz, 1 H), 6.96 (d, J = 2.0 Hz, I H), 6.66 (d, J = 2.0, 8.8 Hz, i H), 6.26 (d, J = 3.2 Hz, 1 H), 1 .67 (s, 9H). MS (ESI) m c (M+H+) 189.2.
[00518] Example 32: 2-iert-Butyl-l-methyl-lH-indol-5-amine
[00519] {2-B romo-4-nitro - pheny 1) -meth l -amine
[00520] To a solution of melhyI-(4-nitro-phenyl)-amine ( 15.2 g, 0.1 mol) in AcOH ( 150 mL) and CHC13 (50 mL) was added Br2 (16.0 g, 0.1 mol) dropwisc at 5 °C. ITie mixture was stirred at 10 °C for l h and then basified with sat. aq. aHCO . The resulting mixture was extracted with EtOAc (100 mL x 3), and the combined organics were dried over anhydrous Na2S04 and evaporated under vacuum to give (2-bromo-4-nitro-phenyl)-methyl-amine (2-bromo-4-nitro-phenyl)-methyl-amine (23.0 g, 99%), which was used in the next step without further purification. Ή NMR (300 MHz, CDCI3) δ 8.37 (d, J = 2.4 Hz, 1 H), 8.13 (dd, J = 2.4, 9.0 Hz, 1 H), 6.58 (d, J = 9.0 Hz, 1 II), 5. 17 (bis, 1 I I), 3.01 (d, J = 5.4 Hz, 3 II).
[00521] [2-(3,3-Dimethyl-biit-l-ynyl)-4-tiitro-phenyl]-methyl-amine
[00522] To a solution of (2-bronio-4-nitro-phenyl)-methyl-amine (22.5 g, 97.4 mmol) in toluene (200 mL) and water ( 100 mL) were added l¾N ( 19.7 g, 1 5 mmol), Pd(PPh3)2Cl2 (6.8 g, 9.7 mmol), Cut (0.7 g, 3.9 mmol) and 3,3-dimethyl-but- l -yne ( 1 .0 g, 195 mmol) successively under N2 protection. The mixture was healed at 70 °C for 3 hours and then cooled down to room temperature. The resulting mixture was extracted with EtOAc ( 100 mL x 3). The combined organic extracts were dried over anhydrous Na2SO, and evaporated under vacuum to give [2-(3,3-dimethyl-but- l -ynyl)-4-nitro-phenyl]-methyl-amine (20.1 g, 94%), which was used in the next step without further purification. !H NMR (400 MHz, CDCb) δ 8.15 (d, J = 2.4 Hz, 1 H), 8.08 (dd, J = 2.8, 9.2 Hz, 1 H), 6.50 (d, J = 9.2 Hz, 1 H), 5.30 (brs, 1 H), 3.00 (s, 311), 1.35 (s, 9H).
[00523] 2-tert-Butyl-l -methyl-5-nitro-lH-indole
[00524] A solution of [2-(3,3-dimethyl-but- 1 -ynyl)-4-nitro-phenyl]-methyl-amine (5.0 g, 22.9 mmol) and TBAF (23.9 g, 91.6 mmol) in THE (50 mL) was heated at reflux overnight. The solvent was removed by evaporation under vacuum and the residue was dissolved in brine (100 mL) and EtOAc ( 100 mL). The organic phase was separated, dried over Na2S()4 and evaporated under vacuum to give 2-tert-butyl- 1 -methyl-5-nitro-lH-indole (5.0 g, 99%), which was used in the next step without further purification. Ή NMR (CDCI3, 400 MHz) 6 8.47 (d, J = 2.4 Hz, 1 H), 8.07 (dd, J = 2.4, 9.2 Hz, 1H), 7.26-7.28 (m, 111), 6.47 (s, IH), 3.94 (s, 3H), 1.50 (s, 911).
[00525] 2-/tr/-But l- l-meth>l-] H-indol-5-aniine
[00526] To a solution of 2-tert-butyl- l-mcthyl-5-nitro-lH-indolc (3.00 g, 13.7 mmol) in MeOH (30 mL) was added Raney Ni (0.3 g) under nitrogen atmosphere. The mixture was stirred under hydrogen atmosphere ( L atm) at room temperature overnight. The mixture was filtered through a Cclitc pad and the filtrate was evaporated under vacuum. The crude residue was purified by column chromatography on silica gel (P.E/EtOAc 20: 1 ) to give 2-lert-butyl-l -methyl- lII-indol-5-amine ( 1 .7 g, 66%). Ή NMR (300 MHz, CDCI3) δ 7.09 (d, J = 8.4 Hz, 1 H), 6.89-6.9 (in, 1 H), 6.66 (dd, J = 2.4, 8.7 Hz, 1 H), 6.14 (d, J = 0.6 Hz, 1 H), 3.83 (s, 3H), 3.40 (brs, 2H), 1.45 (s, 9H); MS (ESI) m/e ( +H+) 203.1.
[00527] Example 33: 2-Cyclopropyl-1H-indol-5-amine
[00528] 2-Bromo-4-nitroaniline
[00529] To a solution of 4-nitro-aniline (25 g, 0.18 mol) in HOAc (150 ml .) was added liquid Br2 (30 g, 0.1 mol) dropwise at room temperature. The mixture was stirred for 2 hours. The solid was collected by filtration and poured into water ( 100 mL), which was basified with sal. aq. NaHCCh to pH 7 and extracted with ElOAc (300 mL x 3). The combined organic layers were dried over anhydrous Na2S0.4 and evaporated under reduced pressure to give 2-bromo-4-nitroaniline (30 g, 80%), which was directly used in the next step.
[00530] 2 -(Cy clopropy lethy nyl)-4 -nitroaniline
[00531] To a dcoxygenated solution of 2-bromo-4-nitroaniline (2. 17 g, 0.01 mmol), ethynyl-cyciopropane ( 1 g, 15 mmol) and Cul (10 mg, 0.05 mmol) in triethylamine (20 mL) was added Pd(PPh3)2Cl2 ( 10 mg, 0.3 mmol) under N2. The mixture was heated at 70 °C and stirred for 24 hours. The solid was filtered off and washed with EtOAc (50 mL x 3). The filtrate was evaporated under reduced pressure, and the residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 10/1 ) to give 2-(cyclopropylethynyl)-4-nitroaniline (470 mg, 23%). Yl NMR (300 MHz, CDC13) δ 8.14 (d, J = 2.7 Hz, I H), 7.97 (dd, J = 2.7, 9.0 Hz, I H), 6.63 (d, ./ = 9.0 Hz, 1 H), 4. 1 (brs, 21 1), 1 .55-1.46 (m, ! H), 0.98-0.90 (m, 2H), 0.89-0.84 (m, 2H).
[00532] N-(2 - ( Cyclopropylethynyl) phenyl) -4- nitrobu t yr amide
[00533] ΓΓο a solution of 2-(cyclopropylethynyl)-4-nitroaniline (3.2 g, 1 .8 mmol) and pyridine (2.47 g, 31.7 mmol) in CH2C¾ (60 mL) was added butyryl chloride (2.54 g, 23.8 mmol) at 0 °C. The mixture was wanned to room temperature and stirred for 3 hours. The resulting mixture was poured into ice-water. The organic layer was separated. The aqueous phase was extracied with CHiCh (30 m L x 3). The combined organic layers were dried over anhydrous Na2S Xi and evaporated under reduced pressure lo give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 10/1 ) to give N-(2-(cyclopropylethynyl)phenyl)-4-nitrobutyramide (3.3 g, 76%). !II NMR (400 MHz, CDC13) δ 8.61 (d, J = 9.2 Hz, 1H), 8.22 (d, J = 2.8 Hz, I II), 8.18 (brs, 1 H), 8.13 (dd, ./ = 2.4, 9.2 Hz, 1H), 2.46 (t, J = 7.2 Hz, 2H), 1.83- 1.76 (m, 211), 1.59- 1.53 (in, 1 H), 1.06 (t, J = 7.2 Hz, 3H), 1.03- 1.01 (m, 2H), 0.91 -0.87 (m, 21-1).
[00534] 2-Cyc]opropyl-5-nilro-lH-indole
[00535] A mixture of N-(2-(cyclopropylethynyl)phenyl)-4-nitrobutyramide (3.3 g, 0.01 mol) and TBAF (9.5 g, 0.04 mol) in THF ( 100 mL) was heated at reflux for 24 hours. The mixture was cooled to the room temperature and poured into ice water. 'Hie mixture was extracted with CH2CI2 (50 m I , x 3). The combined organic layers were dried over anhydrous Na2SC>4 and evaporated under reduced pressure. 'Hie residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 10/1 ) to give 2-cyelopropyl-5-nitro- lH-indole (1.3 g, 64%). ]1-1 NMR (400 MHz, CDC13) δ 8.44 (d, ./ = 2.0 Hz, 11-1), 8.40 (brs, 1H), 8.03 (dd, J = 2.0, 8.8 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1 H), 6.29 (d, J = 0.8 Hz, 1 H), 2.02- 1.96 (m, 111) 1.07- 1.02 (m, 2H), 0.85-0.8 l (m, 2H).
[00536] 2-Cycloprop l- 1 H-iiidol-5-amme
[00537] To a solution of 2-cyclopropyl-5-nitro- I H-indole (1.3 g, 6.4 mmol) in MeOH (30 mL) was added Raney Nickel (0.3 g) under nitrogen atmosphere. The mixture was stirred under hydrogen atmosphere (1 atm) at room temperature overnight, 'Hie catalyst was filtered through a ("elite pad and the filtrate was evaporated under vacuum to give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 5/1 ) to give 2-cycloprop l- l H-indol-5-aminc (510 mg, 56%). !H NMR (400 MHz, CDC1 ) δ 6.89 (d, 7 = 8.4 Hz, 111), 6.50 (d, J = 1.6 Hz, 1H), 6.33 (dd, J = 2.0, 8.4 Hz, 1 H), 5.76 (s, IH), 4.33 (brs, 2H), 1 .91- 1.87 (m, 1 H), 0.90-0.85(m, 211), 0.70-0.66 (m, 2H); MS (ESI) m/e (M+H+) 173.2.
[00538] Example 34: 3-terf-Buty]-lH~indol-5-amine
[00539] 3-/€rt-Butyl-5-nitro-lH-indoIe
[00540] To a mixture of 5-nitro- IH-indole (6 g, 36.8 mmol) and AICI3 (24 g, 0.18 mol) in CH2Cl2 ( 100 ml .) was added 2-hromo-2-methyl-propane (8.1 g, 36.8 mmol) dropwise at O °C. After being stirred at 15 °C overnight, the reaction mixture was poured into ice ( 100 mL). The precipitated salts were removed by filtration and the aqueous layer was extracted with CH2CI2 (30 mL x 3). The combined organic layers were washed with water, brine, dried over a2S0 and concentrated under vacuum to obtain the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 20: 1 ) to give 3-tert-butyl-5 -nitro- I H-indole (2.5 g, 31%). Ή NMR (CD(¾. 400 MHz) δ 8.49 (d, = 1.6 Hz, 1 H), 8.31 (brs, 1 H), 8.05 (dd, J = 2.0, 8.8 Hz, 1 H), 7.33 (d, J = 8.8 Hz, I H), 6.42 (d, J = 1.6 Hz, 1H), 1.42 (s, 9H).
[00541] 3-iifrt-Butyl-l H-indol-5-amine
[00542] To a solution of 3-lert-but l-5-nitro- 1 H-indole (2.5 g, 1 1.6 mmol) in MeOH (30 mL) was added Raney Nickel (0.2 g) under N2 protection. The mixture was stirred under hydrogen atmosphere ( I atm) at 15 °C for 1 hr. The catalyst was filtered off and the filtrate was concentrated under vacuum to dryness. The residue was purified by preparative HLPC to afford 3-tcrt-butyl-lH-indol-5-aminc (0.43 g, 19%). *H NMR (CDC13. 400 MHz) δ 7.72 (brs, 1 H), 7.1 1 (d, ./ = 8.4 Hz, 1H), 6.86 (d, ./ = 2.0 Hz, 1H), 6.59 (dd, J = 2.0, 8.4 Hz, 1 H), 6.09 (d, J = 1.6 Hz, 1 H), 1.37 (s, 9H); MS (ESI) m/e (M+H+) 189. 1.
[00543] Example 35: 2-Pherjy H-indol-5-amine
[00544] 2-Bromo-4-nitroaniline
[00545] To a solution of 4-nitroaniline (50 g, 0.36 mol) in AcOH (500 mL) was added liquid Br2 (60 g, 0.38 mol) dropwise at 5 °C. The mixture was stirred for 30 min at that temperature. rl"he insoluble solid was collected by filtration and poured into EtOAc (200 mL). The mixture was basified with saturated aqueous NaHCOs to pH 7. The organic layer was separated. rHie aqueous phase was extracted with EtOAc (300 mL x 3). The combined organic layers were dried and evaporated under reduced pressure to give 2-bronio-4-nitroanilinc (56 g, 72%), which was directly used in the next step.
[00546] 4-Nitro-2-(phenylethynyl)aniline
[00547] To a deoxygenaled solution oi 2-bromo-4-nitioaniline (2. 17 g, 0.01 nimol), ethynyl-benzene ( 1 .53 g, 0.015 mol) and Cul ( 10 mg, 0-05 mniol) in iriethylaminc (20 mL) was added PdfPPh^C (210 mg, 0.2 nimol) under N2. The mixture was healed at 70 °C and stirred for 24 hours. The solid was filtered off and washed with EtOAc (50 mL x 3). The filtrate was evaporated under reduced pressure and the residue was purified by column chromatography on silica gel (petroleum elher/ethy) acetate = 10/1 ) to give 4-nitn>2-(phenylethynyl)aniline (340 mg, 14%). Ή NMR (300 MHz, CDC13) δ 8.37-8.29 (m, 1 H), 8.08-8.00 (m, 1 1), 7.56-7.51 (m, 211), 7.41 -7.37 (111, 3H), 6.72 (m, 1H), 4.95 (brs, 2H).
[00548] N-(2 -( Pheny lethy nyl)pheny l)-4 -nitrobu tyramide
[00549] To a solution of 4-nitro-2-(phenylethynyl)aniline ( 17 g, 0.07 mmol) and pyridine ( 1 1. 1 g, 0.14 mol) in CH2C12 (100 mL) was added butyryl chloride ( 1 1.5 g, 0.1 mol) at 0 °C. 'Hie mixture was warmed to room temperature and stirred for 3 hours. 'l¾e resulting mixture was poured into ice-water. The organic layer was separated. The aqueous phase was extracted with CH2C12 (30 m L x 3). The combined organic layers were dried over anhydrous Na2S04 and evaporated under reduced pressure. rITie residue was purified by column chromatography on silica gel (petroleum elher/elhyl acetate = 10/1 ) to give N-(2-(phenylethynyl)phenyl)-4-nilrobutyramide (12 g, 55%). Ή NMR (400 MHz, CDC13) δ 8.69 (d, J =9.2 Hz, 1 H), 8.39 (d, 7 =2.8 Hz, 1 H), 8.25-8.20 (m, 2H), 7.58-7.55 (m, 2H), 7.45-7.42 (m, 3H), 2.49 (1, 7 =7.2 Hz, 2H), 1.85- 1.79 (m, 2H), 1 .06 (t, J = 7.2 Hz, 3H).
[00550] 5-Nitro-2-phenyl-lH-indole [00551 ] Λ mixture οΙ' N-(2-(phenylethynyl)phenyl)-4-nitrobutyramide (5.0 g, 0.020 mol) and TBAF (12.7 g, 0.050 mol) in THF (30 niL) was heated at reflux for 24 h. The mixture was cooled to room temperature and poured inlo ice water. The mixture was extracted with CU2CI2 (50 m L x 3). The combined organic layers were dried over anhydrous Na2SC>4 and evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 10/1 ) to give 5-nitro-2-phenyl- l M-indole (3.3 g, 69%). Ή N R (400 MHz, CDCI3) δ 8.67 (s, 111), 8.06 (dd, J = 2.0, 8.8 Hz, 1 1-1), 7.75 (d, J =7.6 Hz, 211), 7.54 (d, ./ =8.8 Hz, I H), 7.45 (t, ./ =7.6 Hz, 2H), 7.36 (t, J = 7.6 Hz, 1H). 6.95 (s, H I).
[00552] 2-Phenyl-lH-indol-5-amine
[00553] To a solution of 5-nitxo-2-phenyl-lH-indole (2.83 g, 0.01 mol) in MeOH (30 mL) was added Raney Ni (510 mg) under nitrogen atmosphere. The mixture was stirred under hydrogen atmosphere ( I atm) at room temperature overnight. The catalyst was filtered through a ('elite pad and the filtrate was evaporated under vacuum to give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 5/1 ) to give 2-phcnyl- l H-indol-5-aminc ( 1 .6 g, 77%) . Ή NMR (400 MHz, CDCI3) 5 7.76 (d, J =7.6 Hz, 2H), 7.39 (t, ./ = 7.6 Hz, 2H), 7.24 (t, J = 7.6 Hz, 1 H), 7.07 (d, J = 8.4 Hz, I H), 6.64 (d, J = 1.6 Hz, 1H), 6.60 (d, J = 1 .2 Hz, 1H), 6.48 (dd, ,/ = 2.0, 8.4 Hz, 111), 4.48 (bis, 2H); MS (ESI) m/e (M+H+) 209.0.
[00554] Example 36: 2- rt-ButyM-fluoro-l H-mdol-5-amine
[00555] 2-Bromo-3-f1uoroanilme
[00556] To a solution of 2-bronio- l -fluorO-3-nitrobenzene ( 1.0 g, 5.0 mmol) in CH3OH (50 niL) was added NiCl2 (2.2 g 1 mmol) and NaliH4 (0.50 g 14 mmol) at 0 °C. After the addition, the mixture was stirred for 5 min. Water (20 niL) was added and the mixture was extracted with EtOAc (20 mL x 3). The organic layers were dried over anhydrous Na2S04 and evaporated under vacuum to give 2-bromo-3-fluoroaniline (600 nig, 70%). Ή NMR (400 MHz, CDCb) 8 7.07-7.02 (m, 1 I I), 6.55-6.49(m, 1 H), 4.22 (br s, 2 H).
[00557] jV-(2-Bromo-3-fluorophenyl)butyramide
[00558] To a solution of 2-bromo-3-nuoroaniline (2.0 g, 1 1 mmol) i n CH2C12 (50 mL) was added butyryl chloride ( 1.3 g, 13 mmol) and pyridine ( 1.7 g, 21 mmol) at 0 °C. The mixture was stirred at room temperature for 24 h. Water (20 mL) was added and the mixture was extracted with CH2C12 (50 111L x 3). The organic layers were dried anhydrous over Na2S0 and evaporated under vacuum to give /V-(2-bromo -fluorophenyl)butyramide (2.0 g, 73%), which was directly used in the next step.
[00559] N-(2-(3,3-Dimethylbut-l-ynyl)-3-nuorop enyl)butyramide
[00560] To a solution of N-(2-bromo-3-fluorophenyl)butyramide (2.0 g, 7.0 mmol) in Et3N (100 mL) was added 4,4-dimethylpenl-2-yne (6.0 g, 60 mmol), Cul (70 nig, 3.8 mmol), and Pd(PPh3)2Cl2 (500 mg) successi vely at room temperature under N2. The mixture was heated at 80 °C overnight. The cooled mixture was filtered and the filtrate was extracted with EtOAc (40 mL x 3). The organic layers were washed with sat. NaCI, dried over anhydrous Na2S04, and evaporated under vacuum. The crude compound was purified by column chromatography on silica gel ( 10% EtOAc in petroleum ether) to give /V-(2-(3,3-dimethylbut-l-ynyl)-3-fIuorophenyl)butyraniide (1. 1 g, 55%). Ή NMR (400 MHz, CDCI3) δ 8.20 (d, J = 7.6, 1 H), 7.95 (s, 1 H), 7.21 (m, 1 H), 6.77 (t, J = 7.6 Hz, 1 H), 2.39 (I, J = 7.6 Hz, 2 H), 1.82- 1.75 (m, 2 H), 1 .40 (s, 9 H), 1 .12 (t, J = 7.2 Hz, 3 H).
[00561] 2-tert-Butyl-4-nuoro-lH-indole
[00562] To a solution of N-(2-(3,3-dimethylbut- 1 -ynyl)-3-fluorophenyl)butyramide (6.0 g, 20 mmol) in DMF ( 100 mL) was added r-BuOK (5.0 g, 50 mmol) at room temperature. 'Ihe mixture was healed at 90 °C overnight before it was poured into water and extracted with EtOAc ( 100 mL x 3). The organic layers were washed with sat. NaCI and water, dried over anhydrous N 2S0 , and evaporated under vacuum to give 2-ferf-butyl-4-fluoro- l H-indole (5.8 g, 97%). l\ l NMR (400 MHz, CDCI3) δ 8.17 (br s, 1 H), 7.1 1 (d, 7=7.2 Hz, 1 H), 7.05-6.99 (m, 1 H), 6.76-6.71 (in, 1 H), 6.34 (m, 1 H), 1.41 (s, 9 H).
[00563] 2--erri-Butyl-4-nuoro-5-nitro-1 H-indole
[00564] To a solution of 2-rerf-butyl-4-f]uon l H-indole (2.5 g, 10 mmol) in H2S04 (30 mL) was added KN03 ( 1 .3 g, 10 mmol) at 0 °C. 'Hie mixture was stirred for 0.5 h at - 10 °C. The mixture was poured into water and extracted with EtOAc (100 mL x 3). The organic layers were washed with sat. NaCI and water, dried over anhydrous Na2S04, and evaporated under vacuum. rl¾c crude compound was purified by column chromatography on silica gel ( 10% EtOAc in petroleum ether) to give 2-/i'/-/-butyl-4-fluoro-5-nilro- IH-indole (900 mg, 73%). Ή NMR (400 MHz, CDC13) δ 8.50 (br s, 1 H), 7.86 (dd, J = 7.6, 8.8 Hz, 1 H), 7.13 (d, J = 8.8 Hz, 1 H), 6.52 (dd, J = 0.4, 2.0 Hz, 1 H), 1.40 (s, 9 H).
[00565] 2-itri-Butyl-4-nuoro-l H-indol-5-aminc
[00566] To a solution of 2-/eri-butyl-4-fluoro-5-nitro- lH-indole (2.1 g, 9.0 mmol) in methanol (50 mL) was added NiCl2 (4.2 g, 18 mmol) and NaBH4 (L0 g, 27 mmol) at 0 °C.
After the addition, the mixture was stirred for 5 min. Water (20 mL) was added and the mixture was extracted with EtOAc (30 mL x 3). The organic layers were washed with sat.
NuCl and water, dried over anhydrous Na2S04, evaporated under vacuum to give 2-/er/-butyl-4-fluoro- lH-indol-5-amine (900 mg, 50%). Ή NMR (300 MHz, CDCl3) δ 7.80 (brs, 1 H), 6.91 (d, J = 8.4 Hz, 1 H), 6.64 (dd, J - 0.9, 2.4 Hz, 1 H), 6.23 (s, 1 H), 1.38 (s, 9 H).
[00567] Example 37: 2,3,4,9-Tetrahydro-1 H-carbazol-6-amine
[00568] 2,3,4,9-Tetrahydro-IH-carbazol-6-amine
[00569] 6-NitiO-2,3,4,9-letrahydro- l -carhazole (0.100 g, 0.462 mmol) was dissolved in a 40 mL scintillation vial containing a magnetic stir bar and 2 mL of ethanol. Tin(H) chloride dihydrate ( 1 .04 g, 4.62 mmol) was added to the reaction mixture and the resulting suspension was heated at 70 °C for 16 h. The crude reaction mixture was then diluted with 15 mL of a saturated aqueous solution of sodium bicarbonate and extracted three times with an equivalent volume of ethyl acetate. rfhe ethyl acetate extracts were combined, dried over sodium sulfate, and evaporated to dryness to yield 2,3,4,9-tetrahydro- lH-carbazol-6-amine (82 mg, 95%) which was used without further purification.
[00570] Example 38: 2-/ert-Butyl-7-nuoro-lH-indol-5-amine [00571 ] 2-Br mo-6-fluoro-4-nitro-phenylamine
[00572] To a solution of 2-fluoro-4-nitro-phenylamine ( 12 g, 77 mmol) in AcOH (50 mL) was added ΙΪΓ2 (3.9 mL, 77 mmol) dropwise at 0 C. The mixture was stirred at 20 °C for 3 h. The reacuon mixture was basilied with sat. aq. NaHCOj, and extracted with EtOAc (100 mL x 3). The combined organics were dried over anhydrous 2S04 and evaporated under vacuum to give 2-bromo-6-fluoro-4-nilro-phenyIamine ( 18 g, 97%). Ή NMR (400 MHz, CDC13) 6 8.22 (m, 1 H), 7.90 (dd, J = 2.4, 10.8 Hz, 1 H), 4.88 (brs, 2 H).
[00573] 2-(3,3-Dimethyl-but-l-ynyl)-6-nuoro-4-nilro-phenylamine
[00574] To a solution of 2-bromo-6-fluoro-4-nitro-phenylamine ( 1 1 g, 47 mmol) in dry I¾N (100 mL) was added Cul (445 mg, 5% mol), Pd(PPh3)2Cl2 (550 mg, 5% mol) and 3,3-dimethyl-but- l -yne (9.6 g, 120 mmol) under N2 protection. The mixture was stirred at 80 U for 10 h. The reaction mixture was filtered, poured into ice (100 g), and extracted with EtOAc (50 mL x 3). The combined organic extracts were dried over anhydrous Na2S04 and evaporated under vacuum to give the crude product, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 50: 1 ) to give 2-(3,3-dimethyl-but- l-ynyl)-6-fluoro-4-nitro-phenyIamine (4.0 g, 36%). Ή NMR (400 MHz, CDClj) δ 8.02 (d, ./ = 1.2 Hz, 1 H), 7.84 (dd, J = 2.4, 10.8 Hz, 1 H), 4.85 (brs, 2 H), 1.36 (s, 9 H).
[00575] V-[2-(3,3-Dimethyl-but-l-ynyl)-6-nuoro-4-nitro-pheny]]-butyramide
[00576] To a solution of 2-(3,3-dimcthyl-but- 1 -ynyl)-6-fluoro-4-nitro-phcnylaminc (4.0 g, 17 mmol) and pyridine (2.7 g, 34 mmol) in anhydrous CH2O2 (30 mL) was added and butyryl chloride (1 .8 g, 17 mmol) dropwise at 0 °C. After stirring for 5 h at 0 °C, the reaction mixture was poured into ice (50 g) and extracted with CH2CI2 (30 mL x 3). The combined organic extracts were dried over anhydrous a2S04 and evaporated under vacuum to give N-[2-(3,3-dimethyl- but-l-ynyl)-6-lluoro-4-nitro-phenyl]-butyramide (3.2 g, 62%), which was used in the next step without further purification. Ή NMR (300 MHz, DMSO) δ 8.10 (dd, J = 1.5, 2.7 Hz, 1 H), 7.95 (dd, J = 2.4, 9.6 Hz, 1 H), 7.22 (brs, 1 H), 2.45 (t, J = 7.5 Hz, 2 H), 1.82 (m, 2 IT), 1.36 (s, 9 H), 1.06 (t, / = 7.5 Hz, 3 I I).
[00577] 2-firrt-ButyL-7-fluoro-5-nitro-lH-indole
[00578] To a solution of W-|2-(3,3-dimcthyl-but- l-ynyl)- 6-nuoro-4-nitro-phenyl]-butyramide (3.2 g, 10 mmol) in DMF (20 niL) was added l-BuOK (2.3 g, 21 mmol) at room temperature. The mixture was heated at 120 °C for 2 g before being cooled down to room temperature. Water (50 m!_) was added to the reaction mixture and the resulting mixture was extracted with CH2Q2 (30 mL x 3). The combined organic extracts were dried over anhydrous Na2SC>4 and evaporated under vacuum to give 2-½ri-butyl-7-f]uoro- 5-nitro-lU-i ndole (2.0 g, 81 %), which was used in the next step without further purification. Ί-1 N R (300 MHz, CDCI3) 6 9.95 (brs, 1 H), 8.30 (d, J = 2.1 Hz, 1 H), 7.74 (dd, J = 1.8, 1 1.1 Hz, 1 H), 6.43 (dd, J = 2.4, 3.3 Hz, 1 H), 1.43 (s, 9 II).
[00579] 2-tert-Butyl-7-nuoro-lH-indol-5-amine
[00580] To a solution of 2-tert-bulyl-7-fluoro- 5-nitro- lH-indole (2.0 g, 8.5 mmol) in MeOH (20 mL) was added Ni (0.3 g) under nitrogen atmosphere. The reaction mixture was stirred under hydrogen atmosphere (1 atm) at room temperature overnight. The catalyst was filtered off through the celite pad and the filtrate was evaporated under vacuum. The crude product was purified by column chromatography on silica gel (petroleum ether/ethyl acetate 100: 1 ) to give 2-/en-butyl-7-fluoro-1 H-indol-5-amine (550 mg, 24%). Ή NMR (300 MHz, CDCI3) 5 7.87 (brs, 1 H), 6.64 (d, / = 1.5 Hz, 1 H), 6.37 (dd, J = 1.8, 12.3 Hz, 1 H), 6.1 1 (dd, J = 2.4, 3.6 Hz, 1 H), 1.39 (s, 9 H). MS (ESI) m/z (M+H+) 207.
[00581] Example 39: 5-Amino-2-iert-but l-IH-iridolc-7-carbonitrile
[00582] 2-Amino-3-(3,3-dimethylbut-l-ynyl)- 5-nitrobenzonitrile
[00583] To a stirred solution of 2-amino-3-bromo-5-nitrobenzonitrile (2.4 g, 10 mmol) in dry Et3N (60 mL) was added Cul (380 mg, 5% mol) and Pd(PPh3)2Cl2 (470 mg, 5% mol) at room temperature. 3,3-dimethyl-but- 1 -yne (2.1 g, 25 mmol) was added dropwise to the mixture at room temperature. The reaction mixture was stirred at 80 °C for 10 h. The reaction mixture was filtered and the filtrate was poured into ice (60 g), extracted with ethyl acetate. The phases were separated and the organic phase was dried over Na2S04. The solvent was removed under vacuum to obtain the crude product, which was purified by column chromatography (2- 10% EtOAc in petroleum ether) to obtain 2-amino-3-(3,3-dimethylbut-l -ynyl)- 5-nitrobenzonitrile (1.7 g, 71 %). Ή NMR (300 MHz, CDC13) S 8.28 (d, J = 2.7 Hz, 1 H), 8.27 (d, ,/ = 2.7 Hz, 1 H), 5.56 (br s, 2 H), 1.37 (s, 9 H).
[00584] 2-it?rf-Butyl-5-nitro-lH-indo]e-7-carbonitrHe
[00585] To a solution of 2-amino-3-(3,3-dimethylbut- 1 -ynyl)- 5-nitrobenzonitrile ( 1.7 g, 7.0 mmol) in THF (35 mL) was added TBAF (9.5 g, 28 mmol) at room temperature. The mixture was heated at reflux overnight. The reaction mixture was cooled and the THF was removed under reduced pressure. Water (50ml) was added to the residue and the mixture was extracted with EtOAc. 'Hie organics were dried over Na2S(>4 and the solvent was evaporated under vacuum to obtain 0.87 g of crude product 2-/er/-butyl-5-nitro- lH-indoIe-7-carbonitrile which was used directly in the next step without purification.
[00586] 5-Amino-2-tert-butyl- lH-indol-7-carbonitrile
[00587] To a solution of crude product 2-fer -butyl-5-nitro- l H-indo!e-7-carbonilrile (0.87 g, 3.6 inmol) in eOH (10 niL) was added NiCI2.6H20 (1.8 g, 7.2 mmol) at -5 nC. 'Ihe reaction mixture was stirred for 30 min, then NaBH4 (0.48g, 14.32 mmol) was added to the reaction mixture at 0 °C. After 5 min, the reaction mixture was quenched with water, filtered and extracted with EtOAc. The combined organic layers were dried over Na2S04 nd concentrated under vacuum to obtain the crude product, which was purified by column chromatography (5-20% EtOAc in petroleum ether) to obtain 5-amino-2-ier/-butyl- l H-indol-7-carbonitrilc (470 mg, 32% over two steps). Ή NMR (400 MHz, CDC13) 6 8.25 (s, 1 I I), 7.06 (d, J =2.4 Hz, 1 H), 6.84 (d, J = 2.4 Hz, 1 H), 6. 14 (d, ./ = 2.4 Hz, 1 H), 3.57 (br s, 2 H), 1.38 (s, 9 H). MS (ESI) m/z: 214 (M+H+).
[00588] Example 40: Methyl 5'amino-2-tert-butyl-1H-indole-7-carboxylate
[00589] 2-tert-Butyl-5-nitro-lH-indole-7-carboxyIic acid
[00590] 2-rm-Butyl-5-niuO- lH-indolc-7-cajbonitrile (4.6 g, 1 mmol) was added to a solution of OH in EtOH (10%, 100 mL) and the mixture was heated at reilux overnight. The solution was evaporaied to remove alcohol, a small amount of water was added, and then the mixture was acidified with dilute hydrochloric acid. Upon standing in the refrigerator, an orange-yellow solid precipitated, which was purified by chromatography on silica gel (15% EtOAc in petroleum ether) to afford 2-ier/-butyl-5-nitro- lH-indole-7-carboxylic acid (4.0 g, 77%). 'H NMR (CDC13, 300 MHz) δ 10.79 (brs, 1 H), 8.66 (s, 1 I I), 8.45(s, 1 I I), 6.57 (s, 1 II), 1.39 (s, 9 H).
[00591] Methyl 2-iert-butyl-5-nitro-lH-indole-7-carboxylate
[00592] SOCI2 (3.6 g, 30mol) was added dropwise to a solution of 2-/m-buty1-5-nilro- 1 H-indole-7-carboxylic acid (4.0 g, 15 mol) and methanol (30 niL) at 0 °C. The mixture was stirred at 80 °C for 12 h. The solvent was evaporated under vacuum and the residue was purified by column chromatography on silica gel (5% EtOAc in petroleum ether) to afford methyl 2-im-butyl-5-nitro- l l l-tndole-7-carboxyla-e {2.95 g, 70%). Ή N R (CDC13, 300 MHz) δ 9.99 (brs, 1 H), 8.70 (d, J = 2.1 Hz, 1 H), 8.65 (d, 7 = 2.1 Hz, 1 H), 6.50 (d, J = 2.4 Hz, 1 H), 4.04 (s, H), 1.44(s, 9H).
[00593] Methyl 5-amino-2-terr-butyl-lH-indole-7-carboxylate
[00594] Λ solution of 2-/eri-butyl-5-nitro- 1 H-indole-7-carboxylale (2.0 g, 7.2 mmo]) and Raney Nickel (200 mg) in CH3OH (50 mL) was stirred for 5 h at the room temperature under H2 atmosphere. The catalyst was filtered off through a celite pad and the filtrate was evaporated under vacuum to give methyl 5-amino-2-tert-butyl- 1 H-indole-7-carboxylaie (1.2 g, 68%) ]H NMR (CDC13.400 MHz) 5 9.34 (brs, 1 H), 7.24 (d, J = 1.6 Hz, 1 H), 7. 10 (s, 1 H), 6.12 (d, J = 1.6 Hz, IH), 3.88 (s, 3H), 1.45 (s, 9H).
[00595] Example 41 : (5-Amino-2-terr-butyl-lH-indol-7-y])methanol
[00596] (2-tert-Butyl-5-nitro-lH-indol-7-yl) methanol
[00597] To a solution of methyl 2-/ert-butyl-5-nitro- lH-indole-7-carboxylate (6.15 g, 22.3 mmol) and dichloromethane (30ml) was added DIBAL-H (1.0 M, 20 ml,, 20 nimol) at 78 °C. The mixture was stirred for 1 h before water (10 mL) was added slowly. The resulting mixture was extracted with EtOAc (120 mL x 3). The combined organic extracts were dried over anhydrous Na2S0 and evaporated under vacuum to give (2-/_,r/-butyl-5-nitro- 1 H-indol-7-yl)methanol (4.0 g, 73%), which was used in the next step directly.
[00598] (5-Amino-2-tert-but l-lH-indol-7-yl)methanol
[00599] A mixture of (2-im-butyl-5-nitro-l l I-indol-7-yl)methanol (4.0 g, 16 mniol) and Raney Nickel (400 nig) in CH3OH ( 100 mL) was stirred for 5 g at room temperature under 112. The catalyst was filtered off through a celite pad and the filtrate was evaporated under vacuum to give (5-amino-2-ieri-but.yl- l H-indol-7-yl)methano! (3.4g, 80%). Ή NMR (CDC13.400 MHz) δ 8.53 (br s, 111), 6.80 (d, J = 2.0 Hz, 1 H), 6.38 (d, J = 1 .6 Hz, 1 H), 4.89 (s, 2 I I), 1.37 (s, 911).
[00600] Example 42: 2-(l- ethylcyclopropyl)-l H-indol-5-amine
[00601] Trimethyl-(l-methyUcyclopropylethynyl)-silane
[00602] To a solution of cyclopropylcthynyl-lriniclhyl-silanc (3.0 g, 22 mmol) in ether (20 mL) was added dropwise «-I3uLi (8.6 mL, 21.7 mol, 2.5 M solution in hexane) at 0 °C. The reaction mixture was stirred at ambient temperature for 24 h before dimethyl sulfate (6.85 g, 54.3 mmol) was added dropwise at -10 °C. The resulting solution was stirred at 10 °C and then at 20 aC for 30 min each. The reaction was quenched by adding a mixture of sat. aq. NH4CI and 25% aq. ammonia (1 :3, 100 mL). The mixture was then stirred at ambient temperature for 1 h. The aqueous phase was extracted with diethyl ether (3 x 50 mL) and the combined organic layers were washed successively with 5% aqueous hydrochloric acid (100 mL), 5% aq. NaHC(¾ solution (100 mL), and water (100 mL). The organics were dried over anhydrous NaS04 and concentrated at ambient pressure. After Iractional distillation under reduced pressure, tximethyl-( l -mclhy]-cyclopropylethynyl)-silane (1 .7 g, 52%) was obtained as a colorless liquid. Ί 1 NMR (400 MHz, CDC13) δ 1.25 (s, 3 H). 0.92-0.86 (in, 2 H), 0.58-0.56 (m, 2 H), 0.15 (s, 9 H).
[00603] 1 - Ethy ny I - 1 -methy 1 -cy clopr opane
[00604] To a solution of triniethyl-( l-methyl-cyclopropylethynyl)-silane (20 g, 0.13 mol) in THF (250 niL) was added TBAF (69 g, 0.26 mol). The mixture was stirred overnight at 20 "C. The mixture was poured into water and the organic layer was separated. 1 Tie aqueous phase was extracted with THF (50 mL). The combined organic layers were dried over anhydrous Na2SO_i and distilled under atmospheric pressure to obtain 1-ethynyl- 1 -methyl-cyclopropane (7.0 g, contained 1/2 THF, 34%). Ή NMR (400 MHz, CDC13) δ 1.82 (s, 1 H), 1 .26 (s, 3 H), 0.90-0.88 (m, 2 I I), 0.57-0.55 (m, 2 H).
[00605] 2-Bromo-4-nitroaniline
[00606] To a solution of 4-nitro-phenylaniine (50 g, 0.36 mol) in AcOH (500 mL) was added Br2 (60 g, 0.38 mol) dropwisc at 5 °C 'lite mixture was stirred for 30 niin at that temperature, 'ihe insoluble solid was collected by filtration and basified with saturated aqueous Nal IC(¾ to pi 1 7. The aqueous phase was extracted with EtOAc (300 mL x 3). The combined organic layers were dried and evaporated under reduced pressure to obtain compound 2-bromo-4-nitroaniline (56 g, 72%), which was directly used in the next step.
[00607] 2-(( l-Methylcyclopropyl)ethyiiyl)-4-nitroaniline
[00608] To a deoxygcnated solution of 2-bromo-4-nitroaniline (430 mg, 2.0 mniol) and 1-elhynyl-1 -methyl-cyclopropane (630 mg, 8.0 mmol) in triethylamine (20 mL) was added Cul (76 mg, 0.40 mmol) and Pd(PPh3)2Cl2 ( 140 mg, 0.20 mmol) under N2. The mixture was heated at 70 °C and stirred for 24 h. The solid was filtered off and washed with EtOAc (50 ml . x 3). The filtrate was evaporated under reduced pressure and the residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 10/1 ) to give 2-(( 1-methylcyclopropyl)ethynyl)-4-nitroaniline (340 mg, 79%). ]H NMR (300 MHz, CDCI3) 8 8. 15-8.14 (m, 1 I I), 7.98-7.95 (m, 1 I I), 6.63 (d, .7 = 6.9 Hz, ! I I), 4.80 (brs, 2 I I), 1.38 (s, 3 H), 1 .04- 1.01 (m, 2 H), 0.76-0.73 (m, 2 H).
[00609] jV-[2-(l-Methyl-cyclopropylethynyl)-4-nitro-phenyl]-butyramide
[00610] To a solution of 2-(( l -methylcyclopropyl)ethynyl)-4-nitroaniline (220 mg, 1.0 mmol) and pyridine ( 160 mg, 2.0 mol) in CH2C12 (20 mL) was added butyryl chloride ( 140 mg, 1 .3 mmol) at 0 "C. The mixture was warmed to room temperature and stirred for 3 h. The mixture was poured into ice-water. The organic layer was separated and the aqueous phase was extracted with CH2C12 (30 mL x 3). The combined organic layers were dried over anhydrous Na2S()4 and evaporated under reduced pressure to obtain N-| 2-( l-methyl-cycIopropyl-ethynyI)-4-nitro-phenyl]-butyramide (230 mg, 82%), which was directly used in the next step.
[00611] 2-(l-Methylcyclopropyl)-5-nitro-lH-indole
[00612] A mixture of jV-[2-(l-mcthyl-cyclopropylethynyl)-4-nitro-phenyl]-butyramide ( 1.3 g, 4.6 mmol) and TBAF (2.4 g, 9.2 mmol) in THF (20 mL) was heated at reflux for 24 h. The mixture was cooled to room temperature and poured into ice water. The mixture was extracted with CH2CI2 (30 mL x 3). The combined organic layers were dried over anhydrous Na2S0 and evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 10/1) to afford 2-(l-methylcyclopropyl)-5-nitro- lH-indole (0.70 g, 71 %). Ή NMR (400 MHz, CDC13) δ 8.56 (bis, I I I), 8.44 (d, ./ = 2.0 Hz, 1 I I), 8.01 (dd, J = 2.4, 8.8 Hz, 1 H), 7.30 (d, J = 8.8 Hz, 1 1-1), 6.34 (d, J = 1.6 Hz, 1 H), 1.52 (s, 3 H), 1 .03-0.97 (m, 2 H), 0.89-0.83 (m, 2 H).
[00613] 2-(l -Methyl-cyclopropyl)- lH-indol-5-ylamine
[00614] To a solution of 2-( l-methylcyclopropyl)-5-nitro- lH-indole (0.70 g, 3.2 niniol) in ElOH (20 mL) was added Raney Nickel (100 mg) under nitrogen atmosphere. The mixture was stirred under hydrogen atmosphere (1 atm) at room temperature overnight. The catalyst was filtered off through a celite pad and the filtrate was evaporated under vacuum. The residue was purified by column chromatography on silica gel (pelroleuiii ether/ethyl acetate = 5/1 ) to afford 2-( I -methyl-cyclopropyl)- 1 11 -indol-5-ylamine ( 170 mg, 28%). Ή NMR (400 MHz, CDC13) δ 7.65 (brs, 1 H), 7.08 (d, J = 8.4 Hz, 1 I I), 6.82 (s, 1 II), 6.57 (d, J = 8.4 Hz, 1 H), 6.14 (s, 1 H), 3.45 (brs, 2 H), 1.47 (s, 3 H), 0.82-0.78 (m, 2 I I), 0.68-0.63 (m, 2 H).
[00615] Example 43: Methyl 2-(5-amino-l H-indol-2-y])-2-methylpropanoate
[00616] Methyl 2,2-dimeth l-3-oxobutanoate
[00617] To a suspension of NaM (42 g, 1.1 mol , 60%) in Π IF (400 ml .) was added dropwise a solution of methyl 3-oxobutanoate (1 16 g, 1.00 mol) in THF ( 100 mL) at 0 °C. The mixture was stirred for 0.5 h at that temperature before Mel (146 g, 1.1 mol) was added dropwise at 0 °C. The resultant mixture was warmed to room temperature and stirred for 1 h. NaH (42 g, 1.05 mol, 60%) was added in portions at 0 °C and the resulting mixture was continued to stir for 0.5 h at this temperature. Mel ( 146 g, 1.05 mo!) was added dropwise at 0 UC. The reaction mixture was wanned to room temperature and stirred overnight. The mixture was poured into ice water and the organic layer was separated. The aqueous phase was extracted with EtOAc (500 mL x 3). The combined organic layers were dried and evaporated under reduced pressure to give methyl 2,2-dimelhyl-3-oxobutanoale (85 g), which was used directly in the next step.
[00618] Methyl 3-chloro-2,2-dimethylbut-3-enoate
[00619] To a suspention of PC15 (270 g, 1.3 mol) in CH2C12 ( 1000 mL) was added dropwise methyl 2,2-dimelhyl-3-oxobutanoale (85 g) at 0 ° , following by addition of approximately 30 drops of dry MF. The mixture was heated at reflux overnight. The reaction mixture was cooled to ambient temperature and slowly poured into ice water. The organic layer was separated and the aqueous phase was extracted with CH2CI2 (500 mL x 3). The combined organic layers were washed with saturated aqueous Nal-ICOj and dried over anhydrous Na2S04. The solvent was evaporated and the residue was distilled under reduced pressure to give methyl 3-chloro-2,2-dimethylbut-3-enoate (37 g, 23%). Ή NMR (400 MHz, CDC13) 6 5.33 (s, 1 H), 3.73 (s, 3 H), 1.44 (s, 6 H).
[00620] 3-Chloro-2,2-d.methylbut-3-enoic acid
[00621] A mixture of methyl 3-chloro-2,2-dimethylbul-3-enoate (33 g, 0.2 mol) and NaOH (9.6 g, 0.24 mol) in water (200 mL) was heated at reflux for 5 h. The mixture was cooled to ambient temperature and extracted with ether. The organic layer was discarded. ITie aqueous layer was acidified with cold 20% MCI solution and extracted ether (200 mL x 3).
The combined organic layers were dried and evaporated under reduced pressure to give 3-chloro-2,2-dimethyl-but-3-enoic acid (21 g, 70%), which was used directly in the next step. Ή NMR (400 MHz, CDC13) δ 7.90 (brs, 1 H), 5.37 (dd, J = 2.4, 6.8 Hz, 2 II), 1.47 (s, 6 H).
[00622] 2,2-Dimethyl-but-3-ynoic acid
[00623] Liquid NH3 was condensed in a 3-neck, 250 niL round bottom flask at -78 °C. Na (3.98 g, 0.173 mol) was added to the flask in portions. The mixture was stirred for 2 h at -78 °C before anhydrous DMSO (20 mL) was added dropwise at - 78 °C. The mixture was stirred at room temperature until no more NH3 was given off. A solution of 3-chloro-2,2-dimethyl-but-3-enoic acid (6.5 g, 43 mmol) in DMSO (10 mL) was added dropwise at -40 °C. The mixture was warmed and stirred at 50 °C for 5 h, then stirred at room temperature overnight. The cloudy, olive green solution was poured into cold 20% HCl solution and then extracted three times with ether. The ether extracts were dried over anhydrous Na2S04 and concentrated to give crude 2,2-dimelhyl-but-3-ynoic acid (2 g), which was used directly in the next step. JII NMR (400 MHz, CDCI3) 6 2.30 (s, 1 H), 1 -52 (s, 6 H).
[00624] Methyl 2,2-dimethylbiit-3-ynoate
[00625] To a solution of diazomethane (-10 g) in ether (400 mL) was added dropwise 2,2-dimethyl-but-3-ynoic acid (10.5 g, 93.7 mmol) at 0 °C. The mixture was warmed to room temperature and stirred overnight. The mixture was distilled under atmospheric pressure to give crude methyl 2,2-dimethylbut-3-ynoate (14 g), which was used directly in the next step. Ή NMR (400 MHz, CDCI3) δ 3.76 (s, 3 H), 2.28 (s, 1 H), 1.50 (s, 6 H).
[00626] Methyl 4-(2-amino-5-nitrophenyl)-2,2-dimethylbut-3-ynoate
[00627] To a deoxygenated solution of compound 2-bromo-4-nkroani)ine (9.43 g, 43.7 mmol), methyl 2,2-dimeLhylbut-3-ynoate (5.00 g, 39.7 mmol), Cul (754 mg, 3.97 mmol) and triethylamine (8.03 g, 79.4 mmol) in loluene/H20 (100/30 mL) was added Pd(PPh3), (6.17 g, 3.97 mmol) under N2. The mixture was heated at 70 °C and stirred for 24 h. After cooling, the solid was filtered off and washed with I.lOAc (50 mL x 3). The organic layer was separated and the aqueous phase was washed with EtOAc (50 mL x 3). The combined organic layers were dried and evaporated under reduced pressure to give a residue, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 10/1) to obtain methyl 4-(2-amino-5-nitrophenyl)-2,2-dimcthylbut-3-ynoate (900 mg, 9%). Ί-1 NMR (400 MHz, CDC¾) 8 8.17 (d, ./ = 2.8 Hz, 1 H), 8. 1 (dd, ./ = 2.8, 9.2 Hz, 1 H), 6.65 (d, ./ = 9.2 Hz, 1 H), 5. 10 (brs, 2 H), 3.80 (s, 3 H), 1.60 (s, 6 11).
[00628] Methyl 4-(2-butyramido-5-nitrophenyl)-2,2-dimethylbut-3-ynoate
[00629] To a solution of methyl 4-(2-amino-5-nitrophenyl)-2,2-dimethylbut-3-ynoate (260 mg, 1.0 mmol) and pyridine ( 160 mg, 2.0 mol) in CH2CI2 (20 mL) was added butyryl chloride ( 140 mg, 1.3 mmol) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 3 h before the mixture was poured into ice-water. The organic layer was separated and the aqueous phase was extracted with CH2CI2 (30 mL x 3). The combined organic layers were dried over anhydrous Na2S04 and evaporated under reduced pressure to obtain methyl 4-(2-butyramido-5-nitrophenyl)-2,2-dimethylbut-3-ynoate (150 mg, 45%), which was used directly in the next step. Ή NMR (400 MHz, CDC13) δ 8.79 (brs, 1 H), 8.71 (d, J = 9.2 Hz, 1 H), 8.24 (d, J = 2.8 Hz, I H), 8.17 (dd, J = 2.8, 9.2 Hz, 1 H), 3.82 (s, 3 H), 2.55 (t, J = 7.2 Hz, 2 H), 1.85- 1.75 (m, 2 H), 1.63 (s, 6 H). 1.06 (t, J = 6.8 Hz, 3 H).
[00630] Methyl 2-methyl-2-(5-nitro-lH-indol-2-yl)propanoate
[00631] To a deoxygenated solution of methyl 4-(2-butyramido-5-nitrophenyl)-2,2-dimethylbut-3-ynoate (1.8 g, 5.4 mmol) in acetonitrile (30 mL) was added Pd(Cl-l3CN)2Cl2 (0.42 g, 1.6= mmol) under N2. The mixture was heated at reflux for 24 h. After cooling the mixture to ambient temperature, the solid was filtered off and washed with EtOAc (50 mL x 3). The fillrate was evaporated under reduced pressure to give a residue, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 30/1) to give methyl 2-methyl-2-(5-nitro-l H-indol-2-yl)propanoale (320 mg, 23%). Ή NMR (400 MHz, C C13) 6 9.05 (brs, 1 H), 8.52 (d, J = 2.0 Hz, 1 H), 8.09 (dd, J = 2.0, 8.8 Hz, 1 H), 7.37 (d, J = 8.8 Hz, 1 H), 6.54 (d, / = 1.6 Hz, 1 H), 3.78 (d, J = 9.6 Hz, 3 H), 1.70 (s, 6 H).
[00632] Methyl 2-(5-amino-l H-indol-2-yl)-2-rnethylpropanoate
[00633] A suspension of methyl 2-methyl-2-(5-nitro- lH-indol-2-yl)propanoate (60 mg, 0.23 mmol) and Raney Nickel (10 mg) in MeOH (5 mL) was hydrogenatcd under hydrogen ( I atm) at room temperature overnight. The catalyst was filtered off through a eel i te pad and the filtrate was evaporated under vacuum to give a residue, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 5/1) to give methyl 2-(5-amino-I H-indol-2-yl)-2-methylpropanoate (20 mg, 38%). Ή NMR (400 MHz, CDCI3) δ 8.37 (br s, I H), 7.13 (d, J = 8.4 Hz, I H), 6.87 (d, J = 2.0 Hz, 1 H), 6.63 (dd, J = 2.0, 8.4 Hz, 1 H), 6.20 (d, J = 1.2 Hz, 1 H), 3.72 (d, J = 7.6 Hz, 3 H), 3.43 (br s, 1 H), 1.65 (s, 6 H); MS (ESI) m/e (M+H+) 233.2.
[00634] Example 44: 2-Isopropyl-I H-indol-5-aniine
[00635] 2-lsopropyl-5-nitro- l H-indol
[00636] Λ mixture of methyl 4-(2-butyraniido-5-nitn)phenyl)-2,2-dimethylbul-3-ynoate (0.50 g, 1.5 mmol) and TB AF (790 mg, 3.0 mmol) in DMF (20 mL) was heated at 70 °C for 24 h. The reaction mixture was cooled to room temperature and poured into ice water. The mixture was extracted with ether (30 mL x 3). The combined organic layers were dried over anhydrous Na2 04 and evaporated under reduced pressure to give a residue, which was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 20/1 ) to give 2-isopropyl-5-nitro- l l-I-indole ( 100 mg, 33%). Ή NMR (400 MHz, CDC13) δ 8.68 (s, 1 H), 8.25 (br s, 1 I I), 8.21 (dd, J = 2.4, 10.0 Hz, 1 H), 7.32 (d, J = 8.8 Hz, 1 H), 6.41 (s, 1 H), 3.07-3. 14 (m, 1 H), 1 .39 (d, J = 6.8 Hz, 6 H).
[00637] 2-Isopropyl- 1 H-indol-5-amine
[00638] A suspension of 2-isopropyl-5-nitro- l H-indole ( 100 mg, 0.49 mmol) and Raney Nickel (10 mg) in McOH (10 mL) was hydrogenatcd under hydrogen (1 atm) at the room temperature overnight, llic catalyst was filtered off through a celite pad and the filtrate was evaporated under vacuum to give a residue, which was purified by column (petroleum ether/ethyl acetate = 5/1) to give 2-isopropy]- l H-indol-5-amine (35 mg, 41%). Ή NMR (400 MHz, CDC13) δ 7.69 (br s, I I I), 7.10 (d, J = 8.4 Hz, 1 H), 6.86 (d, J = 2.4IIz, 1 I I), 6.58 (dd, J = 2.4, 8.8 Hz, 1 H), 6.07 (t, J = 1.2 Hz, 1 H), 3.55 (br s, 2 H), 3.06-2.99 (m, 1 H), 1.33 (d, J = 7.2 Hz, 6 H); MS (ESI) m/e (M+H+) 175.4.
[00639] Example 45: l-(Benzo[d][l,3]dioxol-5-yl)-/V-{2-(l-hydroxy-2-methylpropan-2-yl)-lH-indol-5-yl)cyclopropanecarboxamide
[00640] Triphenyl(2-atninobenzyl)phosphonium bromide
[00641] 2-Aminobenzyl alcohol (60.0 g, 0.487 mol) was dissolved in ucetonitrile (2.5 L) and brought to reflux. Triphenylphosphine hydrobromide (167 g, 0.487 mol) was added and the mixture was heated at reflux for 3 h. The reaction mixture was concentrated to approximately 500 mL and left at room temperature for 1 h. The precipitate was filtered and washed with cold acetoniLrile followed by hexane. The solid was dried overnight at 40 °C under vacuum to give triphenyl(2-aminobenzyl)phosphonium bromide ( 193 g, 88%).
[00642] Triphenyl((ethyl(2-carbamoyl)acetate)-2-benzyl)phosphonium bromide
[00643] To a suspension of triphenyl(2-aminobenzyl)phosphonium bromide (190 g, 0.43 mol) in anhydrous dichloromethane ( 1 L) was added ethyl malonyl chloride (55 ml, 0.43 mol). The reaction was stirred for 3 h at room temperature. The mixture was evaporated to dryness before elhanol (400 mL) was added. The mixture was heated at reflux until a clear solution was obtained. The solution was left at room temperature for 3 h. The precipitate was filtered, washed with cold elhanol followed by hexane and dried. A second crop was obtained from the mother liquor in the same way. In order to remove residual elhanol both crops were combined and dissolved in dichloromethane (approximately 700 ml.) under heating and evaporated. The solid was dried overnight at 50 °C under vacuum to give triphenyl((emyl(2-carbamoyl)acetate)-2-benzyl)-phosphonium bromide ( 139 g, 58%).
[00644] Ethyl 2-(lH-indol-2-yl)acetate
[00645] Ί nphenyl((ethyl(2-carbamoyl)acetate)-2-ben yl)phosphoniun) bromide (32.2 g, 57.3 mmol) was added to anhydrous toluene (150 nit) and the mixture was heated at reflux. Fresh potassium feri-butoxide (7.08 g, 63.1 mmol) was added in portions over 15 minutes. Reflux was continued for another 30 minutes. The mixture was filtered hot through a plug of celite and evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (0-30% ethyl acetate in hexanc over 45 min) to give ethyl 2-( 1 H-indol-2-yl)acetate (9.12 g, 78%).
[00646] tert-Bulyl 2-((ethoxycarbonyl)methyl)-lH-indole-l-carboxylate
[00647] To a solution of ethyl 2-( 1 H-indol-2-yl)acetate (14.7 g, 72.2 mmol) in dichloromethane ( 150 mL) was added 4-dimeLhylaminopyridine (8.83 g, 72.2 mmol) and di-tcrt-butyl carbonate (23.7 g, 108 mmol) in portions. After stirring for 2 h at room temperature, the mixture was diluted with dichloromethane, washed with water, dried over magnesium sulfate and purified by silica gel chromatography (0 to 20% EtOAc in hexane) to give rm-butyl 2-((ethoxycarbonyl)methyl)-lH-indole-l -carboxylate (20.0 g, 91 %).
[00648] .erf-Butyl 2-(2-(ethoxycarbonyI)propan-2-yl)-lH-indole- l-carboxylate
[00649] ieri-Butyl 2-((ethoxycarbonyl)methyl)- lH-indole-l-carboxylate ( 16.7 g, 54.9 mmol) was added to anhydrous THF (100 mL) and cooled to -78 UC. A 0.5M solution of potassium hexamethyldisilazane ( 165 mL, 82 mmol) was added slowly such that the internal temperature stayed below -60 °C. Stirring was continued for 30 minutes at -78 °C. To this mixture, methyl iodide (5.64 mL, 91 mmol) was added. The mixture was stirred for 30 min at room temperature and then cooled to -78 °C. A 0.5 solution of potassium hexamethyldisilazane (210 mL, 104 mmol) was added slowly and the mixture was stirred for another 30 minutes at -78 °C. More methyl iodide (8.6 mL, 137 mmol) was added and the mixture was stirred for 1.5 h at room temperature. 'ITie reaction was quenched with sat. aq. ammonium chloride and partitioned between water and dichloromethane. The aqueous phase was extracted with dichloromethane and the combined organic phases were dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (0 to 20% elhylacetale in hexane) to give fer/-butyl 2-(2-(cthoxycarbonyl)propan-2-yl)- lH-indole- l-carboxylalc (17.1 g, 94%).
[00650] Ethyl 2-(lH-indol-2-yl)-2-methylpropanoate
[00651] /erf-Butyl 2-(2-(ethoxycarbonyl)propan-2-yl)- lH-indole- l-carboxylate (22.9 g, 69.1 mmol) was dissolved in dichloromethane (200 mL) before TFA (70 mL) was added. The mixture was stirred for 5 h at room temperature. The mixture was evaporated to dryness, taken up in dichloromethane and washed with saturated sodium bicarbonate solution, water, and brine. The product was purified by column chromatography on silica gel (0-20% EtOAc in hexane) to give ethyl 2-(l H-indol-2-yI)~2-methylpropanoate (12.5 g, 78%).
[00652] Ethyl 2-methyl-2-(5-n-tro-lH-indol-2-yl)propanoate
[00653] Ethyl 2-(l H-indo]-2-yl)-2-methylpropanoale (1.0 g, 4.3 mmol) was dissolved in concentrated sulfuric acid (6 mL) and cooled to -10 °C (salt/ice-mixture). A solution of sodium nitrate (370 mg, 4.33 mmol) in concentrated sulfuric acid (3 mL) was added dropwise over 30 min. Stirring was continued for another 30 min at - 10 °C. The mixture was poured into ice and the product was extracted with dichloromethane. The combined organic phases were washed with a small amount of sal. aq. sodium bicarbonate. The product was purified by column chromatography on silica gel (5-30% RtOAc in exane) to give ethyl 2-methyl-2-(5-nilro- l H-indol-2-yl)propanoate (0.68 g, 57%).
[00654] 2-Methyl-2-(5-nitro-lH-indol-2-yl)propan-l-ol
[00655] To a cooled solution of LiAlH, ( 1.0 M in THF, 1.1 mL, 1 .1 mmol) in THF (5 raL) at 0 °C was added a solution of ethyl 2-methyl-2-(5-nitro- l H-indol-2-yl)propanoale (0.20 g, 0.72 mmol) in THF (3.4 mL) dropwise. After addition, the mixture was allowed to warm up to room temperature and was stirred for 3 h. The mixture was cooled to 0 °C before water (2 mL) was slowly added followed by careful addition of 15% NaOH (2 mL) and water (4 mL). The mixture was stirred at room temperature for 0.5 h and was filtered through a short plug of celite using ethyl acetate. The organic layer was separated from the aqueous layer, dried over Na2S0 , filtered and evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (ethyl acetate hexane = 1 / 1 ) to give 2-methyl-2-(5-nitro- 1 I I-tndol-2-yl)propan- l -oI (0.098 g, 58%).
[00656] 2-(5-Amino-lH-indol-2-yl)-2-rnethylpropan-l -ol
[00657] To a solution of 2-niethyl-2-(5-nitro- llI-indol-2-yl)propan- l-ol (0.094 g, 0.40 mmol) in ethanol (4 mL) was added tin chloride dihydrate (0.451 g, 2.0 mmol). The mixture was heated in the microwave at 120 °C for 1 h. The mixture was diluted with ethyl acetate and water before being quenched with saturated aqueous Nal lCC . The reaction mixture was filtered through a plug of celite using ethyl acetate. The organic layer was separated from the aqueous layer, dried over Na2S0 , filtered and evaporated under reduced pressure to give 2-(5-amino- lH-indol-2-yl)-2-methylpropan-l -ol (0.080 g, 98%).
[00658] Example 46: 2-(Pyridin-2-yl)- 1 H-indol-5-amine
[00659] 4-Nitro-2-(pyridin~2-ylethynyl)an-line
[00660] 'lb the solution of 2-iodo-4-nitroaniline (3.0 g, 1 1 mmol) in DMF (60m L) and Et3N (60 mL) was added 2-ethynylpyridine (3.0 g, 45 mmol), Pd(PPh3)2Cl2 (600 mg) and Cul (200 mg) under N2. The reaction mixture was stirred at 60 °C for 12 h. The mixture was diluted with water and extracted with dichloromelhane (3 x 100 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SC>4 and concentrated in vacuum. The residue was purified by chromatography on silica gel (5- 10% ethyl acetate/petroleum ether) to afford 4-nitro-2-(pyridin-2-ylethynyl)aniline ( 1 .5 g, 60%). Ή NMR (300 MHz, CDC13) 8 8.60 (s, 1 H), 8.13 (d, J = 2. 1 Hz, 1 H), 7.98 (d, ./ = 1.8, 6.9 Hz, 1 H), 7.87-7.80 (m, 2 H), 7.42-7.39 (m, 1 H), 7.05 (brs, 2 H), 6.80 (d, J = 6.9 Hz, 1 H). [00661 ] 5-Nitro-2-(pyridin-2-yl)-lH-indole O
[00662] To the solution of 4-nitn 2-(pyridin- 2-ylethynyl)aniline ( 1.5 g, 6.3 mmol) in DMF (50 mL) was added t-BuO (1.5 g, 13 mmol). The reaction mixture was stirred at 90 °C for 2 h. The mixture was diluted with water and extracted with dichloromelhane (3 x 50 mL). The combined organic layers were washed with brine, dried over anhydrous Na2S04 and concentrated in vacuum. The residue was purified by chromatography on silica gel (5- 10% ethyl acetate/petroleum ether) to afford 5-nitro-2-(pyridin-2-yl)- l H-indole (1.0 g, 67%yield). Ή N R (300 MHz, d-DMSO) δ 12.40 (s, 1H), 8.66 (d, J = 2, 1 Hz, 1 H), 8.58 (d, 7 = 1 .8 Hz, 1 H), 8.07-7. 1 (m, 3 H), 7.59 (d, / = 6.6 Hz, 1 H), 7.42-7.37 (ni, 2 H).
[00663] 2-(Pyr-din-2-yl)-l H-indol-5-amine
[00664] To a solution of 5-nitro-2-(pyridin-2-yl)-l H-indole (700 nig, 2.9 minol) in ElOH (20 mL) was added SnCh (2.6 g, 12 inmol). The mixture was heated at reflux for 10 h. Water was added and the mixture was extracted with EtOAc (50 mL x 3), The combined organic layers were washed with brine, dried over anhydrous Na2S0 and concentrated in vacuum. The residue was purified by chromatography on silica gel (5- 10% ethyl acelate/petroleum ether) to afford 2-(pyridin-2-yl)- 1 I-I-indol-5-amine ( 120 mg, 20%). 1 H NMR (400 MHz, CDCI3) 5 9.33 (brs, 1 H), 8.55 (dd, J = 1 .2, 3.6 Hz, 1 H), 7.76-7.67 (m, 2 1-1), 7.23 (d, J = 6.4 Hz, 1 H), 7.1 6-7.12 (m, 1 H), 6.94 (d, J = 2.0 Hz, 1 II), 6.84 (d, J = 2.4 Hz, 1 H), 6.71 -6.69 (dd, J = 2.0, 8.4 Hz, 1 H).
[00665] Example 47; 2-1 Pyndin-2- l )- 1 H-indol-5-amine
[00666] [2-(ierr-Butyl-dimethyl-silanyIoxy)-ethyl]-(2-iodo-4-nitro-phenyl)-amine
[00667] To a solution ol 2-iodo-4-nitroaniline (2.0 g, 7.6 nimol) and 2-(teri-butyldimethylsilyloxy)-acetaldehyde (3.5 g, 75% purity, 15 mmol) in methanol (30 mL) was added TFA (1.5 mL) at 0 "C. rltie reaction mixture was stirred at this temperature for 30 mi n before NaCNBH3 (900 mg, ] 5 mmol) was added in portions. ITie mixture was stirred lor 2 h and was then quenched with water. The resulting mixture was extracted with EtOAc (30 mL x 3), the combined organic extracts were dried over anhydrous Na2S04 and evaporated under vacuum, and the residue was purified by chromatography on silica gel (5 % ethyl acetate/petroleum) to afford |2-(ierf-butyl-dimethyl-silanyloxy)-ethyl)-(2-iodo-4-nitro-phenyl)-aniine (800 mg, 25 %). Ή NMR (300 MHz, CDC13) δ 8.57 (d, J = 2.7 Hz, 1 H), 8.12 (dd, 7 = 2.4 ,9.0 Hz, 1 H), 6.49 (d, J= 9.3 Hz, I H), 5.46 (br s, 1 H), 3.89 (t, J = 5.4 Hz, 2 H), 3.35 (q, = 5.4 Hz, 2 H), 0.93 (s, 9 H), 0.10 (s, 6 H).
[00668] 5-{2-[2-(.iirt-Butyl-dimethyl-silanyloxy)-ethylamino]-5-nitro-pheiiy!}-3,3-dimethyl-pent-4-ynoic acid ethyl ester
[00669] To a solution of |2-(/eri-butyl-dimethyl-silanyloxy)-ethyl|-(2-iodo-4-nitro-phenyl)-amine (800 mg, 1.9 mmol) in J¾N (20 mL) was added Pd(PPh3)2Ci2 (300 mg, 0.040 mmol), Cul (76 mg, 0.040 mmol) and 3,3-dimethyl-bul- l -yne (880 mg, 5.7 mmol) successively under N2 protection. The reaction mixture was heated at 80 °C for 6 h and allowed to cool down to room temperature. The resulting mixture was extracted with EtOAc (30 mL x 3). rl"he combined organic extracts were dried over anhydrous Na2SC> and evaporated under vacuum to give 5- {2-[2-(/-,/-/-butyl-dimcthyl-silanyloxy)-cthylamino]-5-ni-ro-phcnyl )-3,3-dimelhyl-pent-4- ynoic acid ethyl ester (700 mg, 82 %), which was used in the next step without further purification. 1 II NMR (400 MHz, CDC13) δ 8.09 (s, 1 H), 8.00 (d, J = 9.2 Hz, 1 H), 6.54 (d, 7 = 9.2 Hz, 1 H), 6.45 (brs, 1 11), 4.17-4.10 (m, 4 H), 3.82 (t, J = 5.6 Hz, 2 H), 3.43 (q, J = 5.6 Hz, 2 H), 2.49 (s, 2 H), 1.38 (s, 6 H), 1.28 (t, J = 7.2 Hz, 3 H), 0.84 (s, 9 I I), 0.00 (s, 6 H). 3-[l-(2-Hydroxy-ethyl)-5-nitro-]H-indol-2-yl]-3-methyl-butyric acid ethyl [00671 ] Λ solution of 5- ( 2-[2-(ii/-i-hutyl-dinieLhyl-silanyloxy)-ethy]aminoJ-5-nitro-phenyl }-3,3- dimethyl-pent-4- ynoic acid ethyl ester (600 mg, 1.34 mmol) and PdCl2(650 mg) in CH3CN (30 mL) was heated at reflux overnight. The resulting mixture was extracted with EtOAc (30 mL x 3). The combined organic extracts were dried over anhydrous Na2S04 and evaporated under vacuum. The residue was dissolved in THF (20 mL) and TBAF (780 mg, 3.0 mmol) was added. The mixture was stirred at room temperature for 1 h, the solvent was removed under vaccum, and the residue was purified by chromatography on silica gel (10% ethyl acetate/petroleum) to afford 3-[i-(2-hydroxy-ethyl)-5-nitro- l H-indol-2-yl]-3-mcthyl-butyric acid ethyl ester (270 mg, 60 %). ]H NMR (300 MHz, CDC13) δ 8.45 (d, J = 2.1 Hz, 1 H), 8.05 (dd, = 2.1 , 9.0 Hz, 1 H), 6.36 (d, ,/ = 9.0 Hz, 1 H), 6.48 (s, 1 H), 4.46 (L, ./ = 6.6 Hz, 2 H), 4.00-3.91 (m, 4 IT), 2.76 (s, 2 H), 1.61 (s, 6 H), 0.99 (t, 7 = 7.2 Hz, 1 H), 0.85 (s, 9 H), 0.03 (s, 6 H).
[00672] 3-[l-(2-Hydroxy-ethyI)-5-nitro-lH.indol-2-yl]-3-methyI-butan-l-oI
[00673] To a solution of 3-[L(2-hydroxy-cthyl)-5-nitro-lH-indol-2-yl]-3-niethyl-butyric acid ethyl ester (700 mg, 2.1 mmol) in THF (25 mL) was added DIBAL-H (1.0 M, 4.2 mL, 4.2 mmol) at -78 °C. The mixture was stirred at room temperature for 1 h. Water (2 mL) was added and the resulting mixture was extracted with EtOAc (15 mL x 3). The combined organic layers were dried over anhydrous Na2S0 and evaporated under vacuum. The residue was purified by chromatography on silica gel (15 % ethyl acetate/petroleum) to afford 3-[ 1 -(2-hydroxy-ethyl)-5-nitro-lH-indol-2-yl]-3-methyl-butan-l-ol (300 mg, 49%). !H NMR (300 MHz, d-DMSO) δ 8.42 (d, J = 1.5 Hz, 1 H), 7.95 (dd, 7 = 1.2, 8.7 Hz, 1 H), 6.36 (d, J = 9.3 Hz, 1 H), 6.50 (s, 1 I I), 5.25 (br s, 1 II), 4.46-4.42 (m, 4 H), 3.69-3.66 (m ,2 II), 3.24-3.21 (m, 2 H), 1.42 (s, 6 H).
[00674] 3-[5-Ainino-l-(2-hydroxy-ethyl)-lH-indol-2-yI]-3-methyI-butan.l-ol
[00675] Λ solution of 3-1 1 -(2-hydroxy-ethyl)-5-nitr -1 H-indol-2-yl]-3-methyl-butan- 1 -ol (300 mg, 1.03 mmol) and Raney Nickel (200 mg,) in CH3OH (30 mL) was stirred for 5 h at room temperature under a H2 atmosphere. The catalyst was filtered through a celite pad and the filtrate was evaporated under vacuum to give a residue, which was purified by preparative TLC to afford 3-[5-amino- l -(2-hydroxy-ethy])- lH-indol-2-yl]-3-methyl-butan- l -ol (70 mg, 26%). Ή NMR (300 MHz, CDClj) 8 7.07 (d, 7 = 8.7 Hz, 1 H), 6.83 (d, 7 = 2.1 Hz, 1 I I), 6.62 (dd, J = 2. 1 , 8.4 Hz, 1 H), 6.15 (s, 1 H), 4.47 (t, J = 5.4 Hz, 2 H), 4.07 (t, J = 5.4 Hz, 2 H), 3.68 (t, J = 5.7 Hz, 2 H), 2.16 (t, J - 5.7 Hz, 2 H), 4.00-3. 1 (in, 4 H), 2.76 (s, 2 H), 1.61 (s, 6 H), 1.42 (s, 6 H).
[00676] Example 48: (m-Butyl 2-(5-amino-lH-indol-2-yl)piperidine-1-carboxylate
[00677] 2-(Piperidin-2-yl)-lH-indo]-5-amine
[00678] 5-Nitjro-2-(pyridin-2-yl)- 1 H-indole ( 1.0 g, 4.2 mmol) was added to HCl/MeOH (2 M, 50 mL). The reaction mixture was stirred at room temperature for I h and the solvent was evaporated under vacuum. P1O2 (200 mg) was added to a solution of the residue in MeOH (50 mL) and the reaction mixture was stirred under hydrogen atmosphere (1 atm) at room temperature for 2 h. The catalyst was filtered through a celite pad and the solvent was evaporated under vacuum to afford 2-(piperidin-2-yl)- lH-indol-5-amine ( ! .0 g), which was directly used in the next step.
[00679] terf-Butyl 2-(5-amino-l H-indol-2-yl)piperidine-l-carboxylate
[00680] To a solution of 2-(piperidin-2-yl)- lH-indol-5-amine (1.0 g) in 1¾N (25 111 L) and THF (25mL) was added 13oc20 (640 mg, 2.9 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was diluted with water and extracted with dichloromethane (3 x 25 ml.-). The combined organic layers were washed with brine, dried over anhydrous a2S04 and concentrated in vacuum. The residue was purified by chromatography on silica gel (5-10% ethyl acetate/petroleum ether) followed by preparative HPI.C to afford ieri-butyl 2-(5-amino- l H-indol-2-yl)piperidine- l -carboxylate (15 nig, 1 % over 2 steps). Ή NMR (400 MHz, CDC13) δ 8.82 (s, 1 H), 7.58 (s, I H), 7.22 (d, J = 8.8 Hz, 1 H), 7.02 (d, ./ = 1.6, 8.0 Hz, 1 1-1), 6.42 (s, 1H), 6.25 (s, 1 H), 3. 1 -3.88 (m, 1 H), 3.12-3.10 (m, 1 II), 2.81-2.76 (m, 1 H), 2.06-1 .97 (m, 4 H), 1.70- 1.58 (m, 2H), 1 .53 (s, 9 H).
[00681] Example 49: 6-amino-lH-indoIe-2-carbonitrile
[00682] (3-Nitrophenyl)hydrazirie hydrochloride
[00683] 3-NitroaniIine (28 g, 0.20 mol) was dissolved in a mixture of 1I20 (40 mL) and 37% HCl (40 mL). A solution of NaN02 (14 g, 0.20 mol) in H20 (60 mL) was added to the mixture at 0 °C, and then a solution of SnCl2.H20 ( 140 g, 0.60 mol) in 37% HCl (100 mL) was added. After stirring at O UC for 0.5 h, the insoluble material was isolated by filtration and was washed with water to give (3-nitraphenyl)hydrazine hydrochloride (28 g, 73%).
[00684] (E)-Ethyl 2-(2-(3-nitrophenyl)hydrazono)propanoate
[00685] (3-Nitrophenyl)hydrazinc hydrochloride (30 g, 0.16 mol) and 2-oxo-propionic acid ethyl ester (22 g, 0.19 mol) were dissolved in ethanol (300 mL). The mixture was stirred at room temperature for 4 h before the solvent was evaporated under reduced pressure to give (£)-ethyl 2-(2-(3-nilrophenyl)hydrazono)pr panoate, which was us d directly in the next step.
[00686] Ethyl 4-nitro- 1 H-indolc-2-carboxytate and ethyl 6-nitro- 1 H-indole-2-carboxylate
[00687] (£)-Ethyl 2-(2-(3-nitrophenyl)hydrazono)propanoate was dissolved in toluene (300 niL) and PPA (30 g) was added. The mixture was heated at reflux overnight and then was cooled to room temperature. ΊΤιβ solvent was decanted and evaporated to obtain a crude mixture that was taken on to the next step without purification ( 15 g, 40%).
[00688] 4-Nitro-lH-indole-2-carboxylic acid and 6-i.itro-lH-indole-2-carboxy.ic acid
[00689] A mixture of ethyl 6-nitro- 1 H-indole-2-carboxylate (0.5 g) and 10 % NaOH (20 mL) was heated at reflux overnight and then was cooled to room temperature. The mixture was extracted with ether and the aqueous phase was acidified with HC1 to pH 1 -2. The insoluble solid was isolated by filtration to give a crude mixture that was taken on to the next step without purification (0.3 g, 68%).
[00690] 4-Nitro-i H-indole-2-carboxamide and 6-nitro- 1 H-indoIe-2-carboxamide
[00691] A mixture of 6-nitro-l H-indoIe-2-carboxylic acid (12 g, 58 mmoi) and SOCl2 (50 mL, 64 mmol) in benzene ( 150 ml.) was healed at reflux for 2 h. The benzene and excess SOC was removed under reduced pressure. 'Ihe residue was dissolved in anhydrous CH2CI2 (250 mL) and NH.1.H2O (22 g, 0.32 mo!) was added dropwise at 0 °C. The mixture was stirred at room temperature for 1 h. The insoluble solid was isolated by filtration to obtain crude mixture (9.0 g, 68%), which was used directly in the next step.
[00692] 4-Nitro-l H-indole-2-carbonitrile and 6-nitro-lH-indole-2-carbonitrile
[00693] 6-Nitro-1 H-indole-2-carboxamide (5.0 g, 24 mniol) was dissolved in CH2C12 (200 mL). EL3N (24 g, 0.24 mol) and (CF3CO)20 (51 g, 0.24 niol) were added dropwise to die mixture at room temperature. The mixture was continued to stir for 1 h and was then poured into water ( 100 mL). The organic layer was separated and the aqueous layer was extracted with EtOAc ( 100 mL x 3). The combined organic layers were dried over Na2SC>4, filtered and concentrated under reduced pressure to obtain crude product which was purified by column chromatography on silica gel to give a impure sample of 4-nilro-l H-indole-2-carbonitrilc (2.5 g, 55%).
[00694] 6-Amino-lH-indole-2-carbomtrile
[00695] A mixture of 6-nitro- l H-indole-2-carbonitrile (2.5 g, 13 mmol) and Raney Nickel (500 mg) in EtOH (50 mL) was stirred at room temperature under H2 (1 aim) for 1 h. Raney Nickel was removed via filtration and the filtrate was evaporated under reduced pressure to give a residue, which was purified by column chromatograpy on silica get to give 6-amino-1 H-indolc-2-carbonitrilc (1.0 g, 49 %). !H NMR (DMSO-i ) δ 12.75 (br s, I H), 7.82 (d, J = 8 Hz, 1 I I), 7.57 (s, 1 H), 7.42 (s, 1 H), 7. 15 (d, J = 8 Hz, 1 H); MS (ESI) m/e (M-f H+) 158.2.
[00696] Example 50: 6-Amino-lH-indole-3-carbon.tr.le
[00697] 6-Nitro-lH-indole-3-carbonitriIe
[00698] To a solution of 6-nitroindole (4.9 g 30 mmol) in DMF (24 mL) and CH3CN (240 mL) was added dropwise a solution of ClS02NCO (5.0 mL) in CH3CN (39 mL) at 0 °C. After addition, the reaction was allowed to warm to room temperature and was stirred for 2 h. The mixture was then poured into ice-water and basified with sat. NaIIC03 solution to pl l 7-8. The mixture was extracted with ethyl acetate. The organics were washed with brine, dried over Na2S(>4 and concentrated to give 6-nitro- 1 H-indole-3-carbonitrile (4.6 g, 82%).
[00699] 6-Amino-l H-indole-3-carbonilrile
[00700] A suspention of e-nitro-l H-indole-S-carborutrile (4.6 g, 25 nimol) and 10% Pd-C (0.46 g) in EtOH (50 ml ,) was stirred under H2 ( l atm) at room temperature overnight. Alter filtration, the filtrate was concentrated and the residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 3/1 ) to give 6-amino- l H-indole-3-carbonitrile (1.0 g, 98%) as a pink solid. XH NMR (DMSO-rf6) δ 1 1.51 (s, 1 H), 7.84 (d, J = 2.4 Hz, 1 H), 7.22 (d, J = 8.4 Hz, 1 H), 6.62 (s, 1 H), 6.56 (d, J = 8.4 Hz, 1 H), 5.0 (s, 2H); MS (ESI) m e (M+H+) 157.1. [00701 ] Example 51: 2-tert-Butyl-lH-indol-6-amine
[00702] /V-o Tolylpivalamide
[00703] To a solution of o-toiylamine (21 g, 0.20 mol) and Et3N (22 g, 0.22 mol) in CH2C12 was added 2,2-dimethyl-propionyl chloride (25 g, 0.21 mol) at 10 °C. After addition, the mixture was stirred overnight at room temperature. 'Fhe mixture was washed with aq. HCl (5%, 80 mL), saturated aq. NaHCC and brine. The organic layer was dried over Na2SC>4 and concentrated under vacuum to give N-o-tolylpivalamide (35 g, 1 %). ]H NMR (300 MHz, CDC13) 6 7.88 (d, J = 7.2 Hz, 1 H), 7.15-7.25 (m, 2 H), 7.05 (t, J = 7.2 Hz, 1 H), 2.26 (s, 3 H), 1.34 (s, 9 H).
[00704] 2-terf-Butyl-lH-indole
[00705] To a solution of W-o-tolylpivalamide (30.0 g, 159 mmol ) in dry THF (100 ml ,) was added dropwise n-BuLi (2.5 in hexanc, 1 0 niL) at 15 ° . After addition, the mixture was stirred overnight at 15 °C. Ί Tie mixture was cooled in an ice-water bath and treated with saturated NH4C1. The organic layer was separated and the aqueous layer was extracted with ethyl acetate. The combined organic layers were dried over anhydrous Na2S04, filtered, and concentrated in vacuum. The residue was purified by column chromatography on silica gel to give 2-im-bulyl- lH-indole (24 g, 88%). Ή NMR (300 MHz, CDCI3) δ 7.99 (br. s, 1 H), 7.54 (d, J = 7.2 Hz, 1 H), 7.05 (d, J = 7.8 Hz, 1 H), 7.06 -7.13 (in, 2 H), 6.26 (s, 1 H), 1.39 (s, 9 II).
[00706] 2-terf-Butyliiidoltne
[00707] To a solution of 2-½r/-bulyl-lH-indole ( 10 g, 48 mmol) in AcOH (40 mL) was added NaBH4 at 10 °C. The mixture was stirred for 20 minutes at 10 °C before being treated dropwise with H20 under ice cooling. The mixture was extracted with ethyl acetate. The combined organic layers were dried over anhydrous N 2S04, filtered, and concentrated under vacuum to give 2-feri-butylindoline (9.8 g), which was used directly in the next step.
[00708] 2-/(irr-butyl-6-nitroindoline and 2-ter/-butyl-5-nitro-lH-indole
[00709] To a solution of 2-ferr-butylindoline (9.7 g) in H2S04 (98%, 80 mL) was slowly added NO3 (5.6 g, 56 mmol) at 0 °C. After addition, the reaction mixture was stirred at room temperature for 1 h. 'ITie mixture was carefully poured into cracked ice, basified with Na2C0 to pH 8 and extracted with ethyl acetate. TTie combined extracts were washed with brine, dried over anhydrous Na2S04 and concentrated under vacuum. 'ITie residue was purified by column chromatography to give 2-fer/-butyl-6-nitroindolinc (4.0 g, 31 % over two steps). lH NMR (300 MHz, CDCI3) δ 7.52 (dd, J = 1.8, 8.1 Hz, 1 H), 7.30 (s, 1 H), 7.08 (d, J = 7.8 Hz, 1 H), 3.76 (t, ./ = 9.6 Hz, 1 H), 2.98 - 3.07 (m, 1 H), 2.82 - 2. 1 (m, 1 H), 0. 1 (s, 9 II).
[00710] l-tert- Buty l-6-nitro- 1 H -indole [00711 ] To a solution of 2-f£?ri-butyl-6-nitroindoline (2.0 g, 9.1 mmol) in 1 ,4-dioxane (20 nit) was added DDQ (6.9 g, 30 mmol) at room temperature. The mixture was heated at reflux for 2.5 h before being filtered and eoncenlraled under vacuum. The residue was puri fied by column chromatography to give 2-/err-butyl-6-nitro- l lI-indole (1.6 g, 80%). Ή NM (300 MHz, CDC ) δ 8.30 (br. s, 1 I I), 8.29 (s, 1 I I), 8.00 (dd, J = 2.1 , 8.7 Hz, 1 I I), 7.53 (d, J = 9.3 Hz, 1 H), 6.38 (s. 1 I I), 1.43 (s, 9 I I).
[00712] 2-tort-Butyl-lH-mdol-6-amine
[00713] To a solution of 2-it?r/-butyl-6-nitro- l H-indole ( 1.3 g, 6.0 mmol) in MeOH ( 10 tnL) was added Raney Nickel (0.2 g). The mixture was hydrogenated under 1 atm of hydrogen at room temperature for 3 h. ITie reaction mixture was filtered and the filtrate was concentrated. The residue was washed with petroleum ether to give 2-ier/-butyI- 1 ll-indol-6-amine ( 1.0 g, 89%). ]H NMR (300 MHz, DMSO-ί ΰ) δ 10.19 (s, 1 H), 6.99 (d, J = 8. 1 Hz, I H), 6.46 (s, 1 H), 6.25 (dd, J = 1.8, 8.1 Hz, 1 H), 5.79 (d, J = 1.8 Hz, 1 H), 4.52 (s, 2 H), 1 .24 (s, 9 I I); MS (ESI) m/e (M+H+) 189.1.
[00714] Example 52: 3-terf-Butyl-lH-indol-6-amine
[00715] 3-/tr/-Bu(yl-6-nitro-l H -indole
[00716] To a mixture of 6-nitroindole (1.0 g, 6.2 mmol), zinc triflale (2.1 g, 5.7 mmol), and TBAI ( 1.7 g, 5.2 mmol) in anhydrous toluene (1 1 niL) was added DIEA (1.5 g, 1 1 mmol) at room temperature under nitrogen. The reaction mixture was stirred for 10 min at 120 °C, followed by the addition of f-bulyl bromide (0.71 g, 5.2 mmol). The resulting mixture was stirred for 45 min at 120 "C. The solid was filtered off and the filtrate was concentrated to dryness. The residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate = 20: 1 ) to give 3- -butyl-6-niuo- l I I-indole (0.25 g, 1 %) as a yellow solid. 'l I-NMR (CDC13) δ 8.32 (d, J = 2. 1 Hz, lH), 8.00 (dd, = 2.1 , 14.4 Hz, 1H), 7.85 (d, ./ = 8.7 Hz, LH), 7.25 (s, 111), 1.46 (s, 9H).
[00717] 3-ftrf -Buty I- 1 H -indol-6-amine
[00718] A suspension of 3-ier/-butyl-6-nitro- 1 H-indole (3.0 g, 14 mmol) and Raney Nickel (0.5 g) was hydrogenated under I I2 ( 1 atm) at room temperature for 3 h. 1 lie catalyst was filtered off and the filtrate was concentrated to dryness. The residue was purified by column on silica gel (petroleum ether/ethyl acetate = 4: 1 ) to give 3-ie/7-butyM H-indol-6-umine (2.0 g, 77%) as a gray solid. fHNMR (CDCb) δ 7.58 (m, 2H), 6.73 (d, J = 1.2 Hz, 1H), 6.66 (s, 1H), 6.57(dd, J = 0.8, 8.6 Hz, 1H), 3.60 (br, 211), 1.42 (s, 9H).
[00719] Example 53: 5-(Trifliioromethyl)- lH-indol-6-amine
[00720] 1 -Methyl-2,4-dinitro-5-(trifluoromethy))benzene
[00721] To a mixture of HNO3 (98%, 30 niL) and H2S04 (98%, 30 mL) was added dropwise 1 -niethyl-3-trifiuoromethy]-benzene (10 g, 63 mmol) at 0 °C. After addition, the mixture was stirred at rt for 30 min and was then poured into ice-water. The precipitate was filtered and washed with water to give l -methyl-2,4-dini-ro-5-trifluoromethyl-ben/.ene (2.0 g, 13%).
[00722] (£)-2-(2,4-Dinitro-5-(trifluoromethyl)phenyl)-N,N-dimethylethenamine
[00723] Λ mixture of 1 -methy1-2,4-dinitro-5-trifluoromethyl-benzerie (2.0 g, 8.0 mmol) and DMA (1.0 g, 8.2 mmol) in DMF (20 niL) was stirred at 100 °C for 30 min. The mixture was poured into ice- water and stirred for 1 h. The precipitate was filtered and washed with water to give (£)-2-(2,4-dinitJO-5-(trinuoromethyl)phenyl)-N,N-dimethylethenamine (2.1 g, 86%).
[00724] 5-(Trifluoromethyl)-lH-indol-6-amine
[00725] A suspension of (^-2-(2,4-dinitro-5-(trifluoromethyl)phenyl)-N/V-dimethylethenamine (2.1 g, 6.9 tnmol) and Raney Nickel (1 g) in cthanol (80 mL) was stirred under H2 (1 atm) at room temperature for 5 h. The catalyst was filtered off and the filtrate was concentrated to dryness. Trie residue was purified by column on silica gel to give 5-(trinuoromethyl)- lH-indol-6-aniine (200 nig, 14%). Ή NMR (DMSO-rfG) 6 10.79 (br s, 1 H), 7.55 (s, 1 H), 7.12 (s, 1 H), 6.78 (s, 1 H), 6.27(s, 1 H), 4.92 (s, 2 H); MS (ESI) ln/e (M+H+): 200.8.
[00726] Example 54: 5-Ethyl-lH-indol-6-amine
[00727] l-(PhenyIsulfonyl)indoline
[00728] To a mixture of DMAP (1.5 g), benzenesuli'onyl chloride (24.0 g, 136 mmol) and indoline ( 14.7 g, 124 mmol) in CH2C12 (200 mL) was added dropwise Et3N (19.0 g, 186 mniol) at ϋ °C. The mixture was stirred at room temperature overnight. The organic layer was washed with water (2x), dried over Na2SC>4 and concentrated to dryness under reduced pressure to obtain l-(phenylsulfonyl)indoline (30.9 g, 96%).
[00729] l-(l-(Phenytsulfonyl)indolin-5-yl)ethanone
[00730] To a suspension of A1C13 (144 g, 1.08 mol) in CH2C12 ( 1070 mL) was added acetic anhydride (54 mL). The mixture was stirred for 15 minutes before a solution of 1 -(phenylsulfonyl)indoline (46.9 g, 0.180 mol) in CH2CI2 (1070 mL) was added dropwise. The mixture was stirred for 5 h and was quenched by the slow addition of crushed ice. The organic layer was separated and the aqueous layer was extracted with C112C12. 'I"he combined organics were washed with saturated aqueous Nal-JCOa and brine, dried over Na2S0 , and concentrated under vacuum to obtain l -( l -(phenylsulfonyl)indolin-5-yl)ethanonc (42.6 g).
[00731] 5-Ethyl-l-(phenylsuironyl)indoline
[00732] To TFA (1600 mL) at 0 °C was added sodium borohydride (64.0 g, 1.69 mol) over 1 h. To this mixture was added dropwise a solution of l-( l-(phenylsulfonyl)indolin-5-yOethanonc (40.0 g, 0.133 mol) in TFA (700 mL) over 1 h. The mixture was then stirred overnight at 25 °C. After dilution with H20 (1600 mL), the mixture was made basic by the addition of sodium hydroxide pellets at 0 °C. The organic layer was separated and the aqueous layer was extracted with CH2C12. The combined organic layers were washed with brine, dried over N 2S04 and concentrated under reduced pressure. The residue was purified by silica column to give 5-ethyl- l-(phenylsulfonyl)indoline (16.2 g, 47% over two steps).
[00733] 5-Ethylindoline
[00734] Λ mixture of 5-elhyl- l-(phenylsulfonyl)indoline (15 g, 0.050 mol) in HBr (48%, 162 mL) was heated at renux for 6 h. The mixture was basified with sat. NaOH lo pH 9 and then it was extracted with ethyl acetate. The organic layer was washed with brine, dried over Na2S04, and concentrated under reduced pressure. The residue was puri ied by silica column to give 5-ethylindoline (2.5 g, 32%).
[00735] 5-Ethyl-6-nitroindoline
[00736] To a solution of 5-elhylindoline (2.5 g, 17 mmol) in H2S04 (98%, 20 mL) was slowly added KN0 (1.7 g, 17 mmol) at 0 °C. The mixture was stirred al O - 10 °C for 10 minutes. 'I'he mixture was then carefully poured into ice, basified with NaOH solution to pH 9, and extracted with ethyl acetate. rI¾e combined extracts were washed with brine, dried over Na2S04 and concentrated to dryness. 'Hie residue was purified by silica column to give 5-ethyl-6-nitroindoline (1.9 g, 58%).
[00737] 5-Ethyl-6-nitro-lH-indole
[00738] To a solution of 5-ethyl-6-nitroindolinc (1.9 g, 9.9 mmol) in CH2C12 (30 mL) was added Mn02 (4.0 g, 46 mmol). The mixture was stirred at ambient temperature for 8 h. The solid was filtered off and the filtrate was concentrated to dryness to give 5-cthyl-6-nitro- l H-indole ( 1. g).
[00739] 5-Ethyl-lH-indol-6-amine
[00740] Λ suspension of 5-ethy1-6-nitro-1 H -indole ( 1.9 g, 10 nimol) and Raney Nickel ( I g) was hydrogenated under 1-12 (1 aim) at room temperature for 2 h. The catalyst was filtered off and the filtrate was concentrated to dryness. The residue was purified by silica gel column to give 5-ethyl-l H-indol-6-amine (760 mg, 48% over two steps). Ή NMR (CDCl.i) 8 7.90 (br s, 1H), 7.41 (s, 1 H), 7.00 (s, 1H), 6.78 (s, 2H), 6.39 (s, 1H), 3.39 (br s, 2H), 2.63 (q, J = 7.2 Hz, 211), 1.29 (t, J = 6.9 Hz, 3H); MS (ESI) m/e (Μ+1-Γ) 161 .1.
[00741] Example 55: Ethyl 6-amino-l H-indole-4-carboxylate
[00742] 2- ethyl-3,5-dinitrobenzoic acid
[00743] To a mixture of HN03 (95%, 80 mL) and H2S04 (98%, 80 niL) was slowly added 2-methylbenzic acid (50 g, 0.37 mol) at 0 °C. After addition, the reaction mixture was stirred below 30 °C for 1.5 h. The mixture then was poured into ice-water and stirred for 15 min. The precipitate was filtered and washed with water to give 2-methyl-3,5-dinitrobenzoic acid (70 g, 84%).
[00744] Ethyl 2-methyl-3,5-dinitrobenzoate
[00745] A mixture of 2-met yl-3,5-dinitrobenzoic acid (50 g, 0.22 mol) in SOCl2 (80 mL) was heated at reflux for 4 h and then was concentrated to dryness. The residue was dissolved in CH2C12 (50 mL), to which EtOH (80 mL) was added and the mixture was stirred at room temperature for 1 h. The mixture was poured into ice-water and extracted with EtOAc (3 x 100 mL). The combined extracts were washed sat. Na2C03 (80 ml ,), water (2 x 100 mL) and brine (100 nil.,), dried over Na2S04 and concentrated to dryness to give ethyl 2-methyl dinitrobenzoate (50 g, 88%)
[00746] (£)-Ethyl 2-(2-(dimethylamino)vinyl)-3,5-dinitrobenzoate
[00747] Λ mixture of ethyl 2-mcthyl-3,5-dinitrobenzoate (35 g, 0.14 niol) and DMA (32 g, 0.27 mol) in DMF (200 niL) was heated at 100 °C for 5 h. The mixture was poured into ice-water and the precipitated solid was filtered and washed with water to give (£)-ethyl 2-(2-(dimethylamino)vinyl)-3,5-dinitrobenzoaie ( 1 1 g, 48%)
[00748] Ethyl 6-amino-lH-indole-4-carboxy]ate
[00749] A mixture of (£ ethyl 2-(2-(diniethylamino)vinyl)-3,5-dinitrobenzoate (1 1 g, 0.037 mol) and SnC!2 (83 g, 0.37 mol) in ethanol was heated at reflux for 4 h. 'Y e mixture was concentrated to dryness and the residue was poured into water and basified using sal. aq. Na2C03 to pH 8. The precipitated solid was filtered and the filtrate was extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with water (2 x 1(X) tuL) and brine (150 mL), dried over Na2S04, and concentrated to dryness. The residue was purified by column on silica gel to give ethyl 6-amino- l H-indole-4-carboxylate (3.0 g, 40%). 'HNMR (DMSO-Je) δ 10.76 (br s, 1 H), 7. ] 1 -7. 14 (m, 2 H), 6.81-6.82 (m, 1 II), 6.67-6.68 (in, 1 H), 4.94 (br s, 2 H), 4.32-4.25 (q, ./ = 7.2 Hz, 2 H), 1.35-1.31 (t, J = 7.2, 3 H); MS (ESI) m/e (M+H+) 205.0.
[00750] Example 56: 5-Fluoro-lH-indol-6-amine [00751 ] l-Fluoro-5- methy 1-2,4 -di nitrobenzene
[00752] To a stirred solution of HN03 (60 ml.,) and H2S04 (80 ml,) was added dropwise ! -fluoro- 3- methyl benzene (28 g, 25 mmol) under ice-cooling at such a rate that the temperature did not rise above 35 °C. The mixture was allowed to stir for 30 min at rt and was then poured into ice water (500 mL). The resulting precipitate (a mixture of l -fluoro-5-mefhyl-2,4-di nitrobenzene and l -fluoro-3-methyl-2,4-dinilrobenzene, 32 g, ca. 7:3 ratio) was collected by filtration and purified by recrystallization front 50 mL isopropyl ether to give pure l-fluoro-5-methy]-2,4-dinitro-benzene as a while solid (18 g, 36%).
[00753] (^ ^-(S-Fluoro^^-dinitrophenyli-i^N-dimethylethenamine
[00754] A mixture of 1 -fluoro-5-methyl-2(4-dinitro-benzene (10 g, 50 mmol), DMA (12 g, 100 mmol) and DMF (50 mL) was heated at 100 °C for 4h. The solution was cooled and poured into water. The precipitated red solid was collected, washed with water, and dried to give (£)-2-(5-fluoro-2,4-dinitiophenyl)- N-dimethylethenamine (8.0 g, 63%).
[00755] 5-Flu
[00756] A suspension of (£)-2-(5-lluoro-2,4-dinitropbenyl)-NN-dimethylethenamine (8.0 g, 31 mmol) and Raney Nickel (8 g) in EtOH (80 mL) was stirred under H2 (40 psi) at room temperature for 1 h. After filtration, the filtrate was concentrated and the residue was purified by column chromatography (petroleum ether/ethyl acetate = 5/1) to give 5-fluoro-lH-indol-6-amine (1.0 g, 16%) as a brown solid. 'HNMR (DMSO-tfc) δ 10.56 (br s, 1 H), 7.07 (d, J = 12 Hz, 1 H), 7.02 (m, I I I), 6.71 (d, J = 8 Hz, III), 6.17 (s, I I I), 3.91 (br s , 211); MS (ESI) m/e (M+H+) 150.1 .
[00757] Example 57: 5-Chloro- I H-indol-6-amine
[00758] 1 -Ch)oro-5-methyl-2,4-dinitrobenzene
[00759] To a stirred solution of HN03 (55 mL) and I I2S0 (79 mL) was added dropwise I -chloro-3-melhylbeni:ene (25.3 g, 200 mmol) under ice-cooling at such a rale that the temperature did not rise above 35 °C. The mixture was allowed to stir for 30 min at ambient temperature and was then poured into ice water (500 mL). The resulting precipitate was collected by filtration and purified by recrystallization to give l -chloro-5-methyl-2,4-dinilrobenzene (26 g, 60%).
[00760] (i:)-2-(5-Chloro-2,4-dinitropheny!)-N,N-dimethylethenamine
[00761] A mixture of l-chloro-5-methyl-2,4-dinitro-benzene ( 1 1.6 g, 50.0 mmol), DMA (1 1.9 g, 100 mmol) in DMF (50 mL) was heated at 100 °C for 4 h. The solution was cooled and poured into water. The precipitated red solid was collected by filtration, washed with water, and dried to give ( ^^-(S-chloro^^-dinitropheny - A'-dimelhylethenamine (9.84 g, 72%).
[00762] 5-Chloro l H-indol-6-amine
[00763] A suspension of (t 2-(5-chloro-2,4-dinitrophenyI)-NN-dimethylethenamine (9.8 g, 36 mmol) and aney Nickel (9.8 g) in EtOH ( 140 mL) was stirred under [-I2 (1 aim) at room temperature for 4 h. After filtration, the filtrate was concentrated and the residue was puri fied by column chromalograph (petroleum ether/ethyl acetate = 10: 1 ) to give 5-chloro-l H-indol-6-amine (0.97 g, 16%) as a gray powder. ΊΐΝΜΚ (CDCh) δ 7.85 (br s, 1 H), 7.52 (s, 1 H), 7.03 (s, 1H), 6.79 (s, 1 H), 6.34 (s, 1 H), 3.91 (br , 1 H); MS (ESI) ni c (M+H+) 166.0.
[00764] Example 58: Ethyl 6-amino-lH-indole-7-carboxylate
[00765] 3-Methyl-2,6-dinitrobenzoic acid
[00766] To a mixture of MN03 (95%, 80 ml.,) and 1 I2S04 (98%, 80 ml,) was slowly added 3 -methyl ben zic acid (50 g, 0.37 inol) at 0 °C. After addition, the mixture was stirred below 30 °C for 1.5 hours. The mixture was then poured into icc-watcr and stirred for 15 min. The precipitate solid was filtered and washed with water to give a mixture of 3-niethyl-2,6-dinitro-benzoic acid and 5-methyl-2,4-dinitrobenzoic acid (70 g, 84%). To a solution of this mixture (70 g, 0.31 mol) in EtOH ( 1 0 mL) was added dropwise SOCl2 (54 g, 0.45 mol). Ihe mixture was heated at reflux for 2 h before being concentrated to dryness under reduced pressure. The residue was partitioned between EtOAc ( 100 mL) and aq. Na2CO.i (10%, 120 mL). The organic layer was washed with brine (50 mL), dried over Na2S04, and concentrated to dryness to obtain ethyl 5-methyl-2,4-dinitrobenzoate (20 g), which was placed aside. The aqueous layer was acidified by HQ to pi 1 2 - 3 and the precipitated solid was fillered, washed with water, and dried in air to give 3-niethyl-2,6-dinilrobenzoic acid (39 g, 47%).
[00767] Ethyl 3-methyl-2,6-dinitrobenzoate
[00768] Λ mixture of 3-niethyl-2,6-diriitrobenzoic acid (39 g, 0.15 mo!) and SOCl2 (80 ml was heated at reflux 4 h. The excess SOCI2 was evaporated off under reduced pressure and the residue was added dropwise to a solution of EtOH ( 100 mL) and EI3N (50 mL). The mixture was stirred at 20 °C for 1 h and then concentrated to dryness. The residue was dissolved in EtOAc (100 mL), washed with Na2C03 (10 %, 40 mL x 2), water (50 mL x 2) and brine (50 mL), dried over Na2S04 and concentrated to give ethyl 3-methyU2,6-dinitrobenzoate (20 g, 53%).
[00769] (/i)-Ethy[ 3-(2-(dimethylamino)viiiyl)-2,6-dmitrobenzoate
[00770] A mixture of ethyl 3-methyl-2,6-dinitrobenzoate (35 g, 0.14 mol) and DMA (32 g, 0.27 mol) in DMF (200 mL) was heated at 100 °C for 5 h. The mixture was poured into ice water. The precipitated solid was filtered and washed with water to give (£)-ethyl 3-(2-(dimethylaniino)vinyl)-2,6-dinitrobenzoate (25 g, 58%). [00771 ] Ethyl 6-amino- 1 H-indole-7-carboxylate
[00772] A mixture of (E)-ethy1 3-(2-(dimethylamino)vinyl)-2,6-dinitrobenzoate (30 g, 0.097 mol) and Raney Nickel ( 10 g) in EtOH (1000 mL) was hydrogenated at room temperature under 50 psi for 2 h. The catalyst was filtered off and the filtrate was concentrated to dryness. The residue was purified by column on silica gel to give ethyl 6-amino-lH-indole-7-carboxylale as an off-white solid (3.2 g, 16%). Ή NMR (DMSO-rfc) 8 10.38 (s, 1 H), 7.42 (d, J = 8.7 Hz, 1 H), 6.98 (t, / = 3.0 Hz, 1 H), 6.65 (s, 2 H), 6.48 (d, J = 8.7 Hz, 1 H), 6.27-6.26 (m, 1 H), 4.38 (q, 7= 7.2 Hz, 2 H), 1 .35 (t, J= 7.2 Hz, 3 H).
[00773] Example 59: Ethyl 6-amino- l H-indole-5-carboxylate
[00774] (E)-EthyE 5-(2-(dimethylamino)vinyl)-2,4-din robenzoate
[00775] A mixture of ethyl 5-methyl-2,4-dinilrobenzoate (39 g, 0.15 mol) and DMA (32 g, 0.27 mol) in DMF (200 mL) was heated at 100 °C lor 5 h. 'I"he mixture was poured into ice water and the precipitated solid was filtered and washed with water to afford (£)-clhyl 5-(2-(dimethylamino)vinyl)-2,4-dinitrobenzoate ( 15 g, 28%).
[00776] Ethyl 6-amino-l H-mdole-5-carboxyIate
[00777] A mixture of ( ethyl 5-(2-(dimethylamino)vinyl)-2,4-dinitrobenzoate (15 g, 0.050 mol) and Raney Nickel (5 g) in LtOH (500 mL) was hydrogenated at room temperature under 50 psi of hydrogen for 2 h. Ί Tie catalyst was filtered off and the filtrate was concentrated to dryness. The residue was purified by column on silica gel to give ethyl 6-amino-lH-indole-5-carboxylate (3.0 g, 30%). Ή NMR (DIVISOR) f> 10.68 (s, 1 H), 7.99 (s, 1 11), 7.01-7.06 (m, 1 II), 6.62 (s, 1 I I), 6.27-6.28 (m, 1 11), 6.16 (s, 2 I I), 4.22 (q, J = 7.2 Hz, 2 H), 1.32- 1.27 (t, J = 7.2 Hz, 3 H).
[00778] Example 60: 5-i(?rf-Butyl-l H-indol-6-amine
[00779] 2-terf-Butyl-4-methylphenyl diethyl phosphate
[00780] To a suspension of NaH (60% in mineral oil, 8.4 g, 0.21 mol) in Ti ll7 (200 mL) was added dropwise a solution of 2-ierf-butyl-4-methylphenol (33 g, 0.20 mol) in THE7 ( 100 mL) at 0 °C The mixture was stirred at 0 UC lor 15 min and then phosphorochloridic acid diethyl ester (37 g, 0.21 mo!) was added dropwise at 0 °C. After addition, the mixture was stirred at ambient temperature for 30 min. The reaction was quenched with sat. NH4CI (300 mL) and then extracted with 1¾0 (350 mL x 2). The combined organic layers were washed with brine, dried over anhydrous Na2SC>4, and then evaporated under vacuum to give 2-teri-butyl-4-methylphenyl diethyl phosphate (contaminated with mineral oil) as a colorless oil (60 g, - 100%), which was used directly in the next step.
[00781] l-tert-Butyl-3-methylbenzene
[00782] To NUi (liquid, 1 00 mL) was added a solution of 2-rerf-butyl-4-methy]pheny] diethyl phosphate (60 g, crude from last step, about 0.2 mol) in l¾0 (anhydrous, 500 mL) at -78 °C under N2 atmosphere. Lithium metal was added to the solution in small pieces until the blue color persisted. The reaction mixture was stirred at -78 °C for 15 min and then was quenched with sat. NH4CI until the mixture turned colorless. Liquid ΝΙ 3 was evaporated and the residue was dissolved in water. The mixture was extracted with Et20 (400 mL x 2). The combined organics were dried over NajSOa and evaporated to give l -fi!rf-butyl-3-methyl benzene (contaminated with mineral oil) as a colorless oil (27 g, 91 %), which was used directly in next step.
[00783] 1 -½rt-Butyl-5-met y 1-2,4-dinitrobenzene and l-fer<-buty !-3-methyl-2,4-dinitro-benzene
[00784] To I INO3 (95%, 14 mL) was added H2S04 (98 %, 20 mL) at 0 °C and then Uert-butyl-3-methylbenzene (7.4 g, -50 mniol, crude from last step) dropwise to the with the temperature being kept below 30 °C. The mixture was stirred at ambient temperature for 30 min, poured onto crushed ice (100 g), and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with water and brine, before being evaporated to give a brown oil, which was purified by column chromatography to give a mixture of l -/er/-butyl-5- nu;thyl-2,4-dinilrobenzene and 1 -fm-buiy1-3-methyl-2,4-dinitrobenzene (2: 1 by NMR) as a yellow oil (9.0 g, 61%).
[00785] (£,')-2-(5-iiri-Butyl-2,4-dinitrophenyl)-/VJJV-dimethylethenainine
[00786] A mixture of l -im-butyl-5-methyl-2,4-dinitrobenzenc and l -te rt-butyI-3-methyl-2,4-dinitrobenzene (9.0 g, 38 mmol, 2: 1 by NMR) and DMA (5.4 g, 45 mmol) in DMF (50 ml ,) was heated at reflux for 2 h before being cooled to room temperature. The reaction mixture was poured into water-ice and extracted with EtOAc (50 niL x 3). The combined organic layers were washed with water and brine, before being evaporated to give a brown oil, which was purified by column to give (E>2-(5-fm-butyl-2,4-dinitrophenyl)-/V,N-dimethylelhen-amine (5.0 g, 68%).
[00787] 5-tert-Buty[-lH-indol-6-amine
[00788] A solution of ( i^-2 5-fer but l-2,4-diniuΌphen l)-N,Λ'-dimethylethen-aιnine (5.3 g, 18 mmol) and tin (II) chloride dihydralc (37 g, 0.18 mol) in ethanol (200 mL) was heated at reflux overnight. The mixture was cooled to room temperature and the solvent was removed under vacuum. The residual slurry was diluted with water (500 mL) and was basifed with 10 % aq. Na2C03 to pH 8. The resulting suspension was extracted with ethyl acetate (3 x 100 mL). The ethyl acetate extract was washed with water and brine, dried over Na2SC>4, and concentrated. The residual solid was washed with CH2CI2 to afford a yellow powder, which was purified by column chromatography to give 5-fert-butyl-l H-indol-6-amine (0.40 g, 12%). Ή NMR (DMSO.rf6) δ 10.34 (br s, 1 H), 7.23 (s, 1 H), 6.92 (s, 1 I I), 6.65 (s, 1H), 6.14 (s, 1 H), 4.43 (br s, 2 H), 2.48 (s, 9 H); MS (LSI) m e (M+H+) 189.1.
[00789] General Procedure IV: Synthesis of acylaminoindoles
[00790] One equivalent of the appropriate carboxylic acid and one equivalent of the appropriate amine were dissolved in NN-dimethylformamide (DMF) containing triethylamine (3 equivalents). O^V-Azabenzotriazol- l -y -NN.N'.N'-letrainethyluronium hexafluorophosphate (I IATU) was added and the solution was allowed to stir. The crude product was purified by reverse-phase preparative liquid chromatography to yield the pure product.
[00791] Example 61 : /V-(2-iert-Butyl-lH-indol-5-yl)-l-(4-methoxyphenyl)-cyclopropanecarboxamide
[00792] 2-ierr-Butyl- 1 H-indol-5-amine ( 19 nig, 0.10 mmol) and 1 -(4-methoxyphenyl)-cyclopropanecarboxylic acid ( 19 mg, 0. 10 mmol) were dissolved in N^V-dimethylformamide ( 1.00 niL) containing triethylamine (28 μΙ , 0.20 mmol). 0-(7-Azabenzotriazol- 1 -yl)-N,N,/V",N-tetramethyluronium hexafluorophosphate (42 mg, 0.1 1 mmol) was added to the mixture and the resulling solution was allowed to stir for 3 hours. The crude reaction mixture was filtered and purified by reverse phase HPLC. 12SI-MS m/z calc. 362.2, found 363.3 (M+l)+; Retention time 3.48 minutes.
[00793] General Procedure V: Synthesis of acylaminoindoles
[00794] One equivalent of the appropriate carboxylic acid was placed in an oven-dried flask under nitrogen. A minimum (3 equivalents) of thionyl chloride and a catalytic amount of and NN-dimcthylformamidc were added and the solution was allowed to stir for 20 minutes at 60 °C. The excess thionyl chloride was removed under vacuum and the resulting solid was suspended in a minimum of anhydrous pyridine. This solution was slowly added to a stirred solution of one equivalent the appropriate amine dissolved in a minimum of anhydrous pyridine. The resulting mixture was allowed to stir for 15 hours at 1 10 °C. 'Ihe mixture was evaporated to dryness, suspended in dichloromethane, and then extracted three times with IN HC1. The organic layer was then dried over sodium sulfate, evaporated to dryness, and then purified by column chromatography.
[00795] Example 62: Ethyl 5-(l-(benzo[d][l ,3]dtoxol-5-yl)cyclopropanecarboxamido)-lH-indole-2-carboxylate (Compd. 28)
[00796] l -Benzo[ 1 ,3]dioxol-5-yl-eyclopropanecurboxylic acid (2.07 g, 10.0 mmol) was dissolved in thionyl chloride (2.2 niL) under N2. V,/V-dimethylformaiiiide (0.3 mL) was added and the solution was allowed to stir for 30 minutes. The excess thionyl chloride was removed under vacuum and the resulting solid was dissolved in anhydrous dichloromethane ( 15 mL) containing triethylamine (2.8 mL, 20.0 mmol). Ethyl 5-amino- l l l-indole-2-carboxylate (2.04 g, 10.0 mmol) in 15 mL of anhydrous dichloromethane was slowly added to the reaction. The resulting solution was allowed to stir lor 1 hour. ITie reaction mixture was diluted to 50 mL with dichloromethane and washed three times with 50 mL of I N HC1, saturated aqueous sodium bicarbonate, and saturated aqueous sodium chloride. The organic layer was dried over sodium sulfate and evaporated to dryness to yield ethyl 5-(l -(benzo[d][l ,3]dioxol-5-yl)cyclopropanecarboxaniido)- lH-indole-2-carboxylate as a gray solid (3.44 g, 88 %). ESI-MS m/z calc. 392.4; found 393.1 (M+l)+ Retention time 3. 17 minutes. Ή N R (400 MHz, DMSO-d6) 6 11.80 (s, 111), 8.64 (s, 1H), 7.83 (m, 1 H), 7.33-7.26 (m, 2H), 7.07 (m, 1H), 7.02 (m, 1H), 6.96-6.89 (m, 2H), 6.02 (s, 2H), 4.33 (q, J = 7.1 Hz, 2H), 1.42- 1.39 (m, 2H), 1.33 (t, J = 7.1 Hz, 311), 1.06-1.03 (in, 2H).
[00797] Example 63: l-(Benzo[i-][l,3]dioxol-S-yl)-iV-(2-tert-butyl-1H-indol-5-yl)cyclopropanecarboxamide
[00798] 1 -Benzol 1 ,3]dioxol-5-yl-cyclopropanecarboxylic acid (1.09 g, 5.30 mmol) was dissolved in 2 mL of thionyl chloride under nitrogen. A catalytic amount (0.3 mL) of N,N-dimethylfonnamide (DMF) was added and the reaction mixture was stirred for 30 minutes. The excess thionyl chloride was evaporated and the resulting residue was dissolved in 15 mL of dichloromethane. This solution was slowly added to a solution of 2-rerf-butyl-lH-indol-5-amine (1.0 g, 5.3 mmol) in 10 mL of dichloromethane containing triethylamine ( 1.69 mL, 12.1 mmol). The resulting solution was allowed to stir for 10 minutes. The solvent was evaporated to dryness and the crude reaction mixture was purified by silica gel column chromatography using a gradient of 5-50 % ethyl acetate in hexanes. The pure fractions were combined and evaporated to dryness to yield a pale pink powder ( 1.24 g 62%). ESI-MS in/z calc. 376.18, found 377.3 (M+ l)+. Retention time of 3.47 minutes. Ή NMR (400 MHz, DMSO) 6 10.77 (s, 111), 8.39 (s, 111), 7.56 (d, J = 1.4 Hz, 1H), 7.15 (d, J = 8.6 Hz, 111), 7.05 - 6.87 (m, 4H), 6.03 (s, 3H), 1.44 - 1.37 (m, 2H), 1.33 (s, 9H), 1.05- 1.00 (m, 2H).
[00799] Example 64: l-(Benzo[d]tl,3]dioxol-5-yl)- -(l -methyl-2-(l-methylcyclopropyl)-lH-mdol-5-yl)cyclopropanecarboxamide
[00800] 1 -Melhyl-2-( 1 -methykyclopropyl}- 1 /-indol-5-amine (20.0 mg, 0. 100 mmol) and l-(benzo|tf]| l ,3]dioxol-5-yl)cyclopropanecarboxylic acid (20.6 mg, 0.100 mmol) were dissolved in NN-dimethylformamide ( 1 mL) containing triclhylamine (42.1 \ih, 0.300 mmol) and a magnetic stir bar. 0-(7-Azabenzotriazol-l-yl)-N,N,N',/V'-tetraniethyluronium hexafluorophosphatc (42 mg, 0.1 1 mmol) was added to the mixture and the resulting solution was allowed to stir for 6 h at 80 °C. The crude product was then purified by preparative HPLC utilizing a gradient of 0-99% acelonitrile in water coniaining 0.05% trifluoroacelic acid to yield l-(benzolf/][ l ,3]dioxol-5-yl)-N-(l -methyl-2-( l -methykyclopropyl)- lH-indol-5-yl)cyclopropanccarboxamidc. ESI-MS m z calc. 388.2, found 389.2 (M+l)+. Retention time of 3.05 minutes.
[00801] Example 65: l-tBenzolt Jtl^ldioxol-S-yli-iV-d^-dimet yl-Z^-dihydro-lH-pyrrolo[l,2-alindol-7-yl)cyclopropanecarboxamide
[00802] l, l-Dimelhyl-2,3-dihydro- l W-pyrrolo[l,2-fl]indol-7 -amine (40.0 mg, 0.200 mmol) and l-(benzo [<■/][ l ,3]dioxol-5-yl)cyclopropanecarboxylic acid (41 ,2 mg, 0.200 mmol) were dissolved in N,/V-dimelhylformamide ( 1 mL) containing triethylamine (84.2 lL, 0.600 mmol) and a magnetic stir bar. 0-(7-Azabenzouiazol- l-yl)-NN,N,N'-tetramethyluronium hexafluorophosphate (84 mg, 0.22 mmol) was added to the mixture and the resulting solution was allowed to stir for 5 minutes at room temperature. The crude product was then purified by preparative HPLC uti lizing a gradient of 0-99% acetonitrile in water containing 0.05% trifluoroacetic acid to yield l -(benzol(/J[ l,3jdioxol-5-yl)-N-(l , l -dimethyl-2,3-dihydro- l i/-pyrrolol l ,2-a]-indol-7-yl)cyclopropanecarboxaniide. ES1-MS m/z calc. 388.2, found 389.2 ( + l)+. Retention time of 2.02 minutes. *H NMR (400 MHz, DMSO-d6) δ 8.41 (s, 1 H), 7.59 (d, J = 1.8 Hz, 1 H), 7.15 (d, J = 8.6 Hz, 1 H), 7.06 - 7.02 (m, 2H), 6.96 - 6.90 (in, 2H), 6.03 (s, 2H), 5.98 (d, J = 0.7 Hz, 1 H), 4.06 (t, J = 6.8 Hz, 2H), 2.35 (t, J = 6.8 Hz, 2H), 1 .42-1.38 (in, 2H), 1 .34 (s, 6H), 1.05- 1.01 (m, 211).
[00803] Example 66: Methyl 5-(l-(benzo[if][l ,3]dioxol-5-yl)cyclopropanecarboxamido)-2-/eri-butyl-l -indole-7-carboxylate
[00804] l-(Benzo[r/J[ l ,3 ]dioxol-5-yl)cyclopropanecarbonyl chloride (45 nig, 0.20 mmol) and methyl 5-aniino-2-ier/-bulyl- l //-indole-7-carboxylate (49.3 mg, 0.200 mmol) were dissolved in N,N-diniethylformamide (2 niL) containing a magnetic stir bar and triethylamine (0.084 mL, 0.60 mmol). The resulting solution was allowed to stir for 10 minutes at room temperature. The crude product was then purified by preparative HPLC using a gradient of 0-99% acetonitrile in water containing 0.05% lrifluoroacetic acid to yield methyl 5-(l -(benzoli/][l ,3]dioxol-5-yl)cyclopropanecarbox-amido)-2-i£,r/-butyl- li -indole-7-carboxylate. ESI-MS m/z calc. 434.2, found 435.5. (M+ l )+. Retention time of 2. 12 minutes.
[00805] Example 67: l -(Benzo[d][l,3]dioxol-5-yl)-/V-(2-(l-hydroxy-2-methylpropan-2-yl)-1 H-indol-5-yl)cyclopropanecarboxamide
[00806] To a solution of l -(benzo|d| | l,3Jdioxol-5-yl)cyclopropanecarboxy]ic acid (0.075 g, 0.36 mmol) in acetonitrile (1.5 mL) were added H13TU (0. 138 g, 0.36 mmol) and Et3N ( 152 μΙ.., 1.09 mmol) at room temperature. The mixture was stirred al room temperature for 10 minutes before a solution of 2-(5-amino- l H-indol-2-yl)-2-methylpropan- l -ol (0.074 g, 0.36 mmol) in acetonitrile (1 .94 mL) was added. After addition, the reaction mixture was stirred at room temperature for 3 h. The solvent was evaporated under reduced pressure and the residue was dissolved in dichloromethane. 'Hie organic layer was washed with 1 N HQ (1 x 3 ml-) and saturated aqueous NallC(>;(l x 3 ml-). The organic layer was dried over Na2S04, filtered and evaporated under reduced pressure. The crude material was purified by column chromatography on silica gel (ethyl acetate/hcxane = 1/1) to give 1-(benzo|dltl,3Jdioxol-5-yl)-N-(2-(l-hydroxy-2-methylpropan-2-yl)-lH-indol-5-y cyclopropanecarboxamide (0.11 g, 75%). Ή NMR (400 MHz, MSO-d6) δ 10.64 (s, 1H), 8.38 (s, III), 7.55 (s, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.04-6.90 (m, 4H), 6.06 (s, 1H), 6.03 (s,2H),4.79 (t,J = 2.7 Hz, 1H), 3.46 (d, J =0.0 Hz, 2H), 1.41-1.39 (m, 2H), 1.26 (s,6H), 1.05-1.02 (m, 211).
[00807] Example 67: l-(Benzo[i/][l,3]dioxol-S-yl)-/V-(2,3,4,9-tetrahydro-lH-carbazol-6-y l)cyclopropanecarboxami de
[00808] 2,3,4.9-Tetrahydro- 1 H-carbazol-6-aminc (81.8 mg, 0.439 mmol) and 1 -(benzofiill l,3]dioxol-5-yl)cyclopropanecarboxylic acid (90.4 mg, 0.439 mmol) were dissolved in acetonitrile (3 mL) containing diisopropylelhyiamine (0.230 niL, 1.32 mmol) and a magnetic stir bar. C?-(7-Azabenzotriazol- l-yl)-N,MA,N'-telramethyluronium hex fluorophosphate (183 mg, 0.482 mmol) was added to the mixture and the resulting solution was allowed to stir for 16 h at 70 °C. The solvent was evaporated and the crude product was then purified on 40 g of silica gel utilizing a gradient of 5-50% ethyl acetate in hexanes to yield l-(benzo[rf|[l,3]dioxol-5-yi)-N-(2,3,4,9-tetrahydro-l /-carbazol-6-y cyclopropanecarboxamide as a beige powder (0.115 g, 70%) after drying. ESI-MS m/z calc.374.2, found 375.3 (M+1)+. Retention time of 3.43 minutes. Ή NMR (400 MHz, DMSO-i/6) 510.52 (s, 111), 8.39 (s, lH), 7.46(d,./= 1.8 Hz, 1H), 7.10 - 6.89 (m, 5H), 6.03 (s, 2H), 2.68 - 2.65 (m, 2H), 2.56 - 2.54 (in, 2H), 1.82 - 1.77 (m, 4H), 1.41 - 1.34 (m, 2H), 1.04-0.97 (m, 2H).
[00809] Example 69: tert-Butyl 4-(5-(l-(benzo[i/][l,3]dioxol-5.y[ cyclopropanecarbox-amido)-l -indol-2 yl)piperidine-l-carboxy!ate
[00810] l -( enzo[f/][ l ,3]diox()]-5-yl)cyc]opropanccarbonyl chloride (43 mg, 0. 19 nimol) and teri-butyl 4-(5-amino-l //-indol-2-yl)piperidine- 1 -carboxylase (60 nig, 0.19 mmol) were dissolved in dichloromethane ( 1 mL) containing a magnetic stir bar and triethylamine (0.056 mL, 0.40 mmol). The resulting solution was allowed to stir for two days at room temperature. The crude product was then evaporated to dryness, dissolved in a minimum of N,N-dimethylformamide, and then purified by preparative 11PLC using a gradient of 0-99% acetonitrile in water containing 0.05% trifluoroacetic acid to yield ieri-buty! 4-(5-( l -(benzo[rf][ 1 ,3 |dioxol-5-yl)cyclopropanecarboxamido)- l /-indol-2-yl)piperidine- 1 -carboxylatc. ES1-MS mJz calc. 503.2, found 504.5. ( + 1 )+. Retention time of 1.99 minutes. [00811 ] Example 70: Ethyl 2-(5-(l-(benzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamido)- 1 H-indoI-2-yl)propanoate
[00812] terf-Butyl 2-(l -ethoxy-l-oxopropan-2-yl)-lH-indo]e-l-carboxylate
[00813] ter(-\iuty\ 2-(2-ethoxy-2-oxoethyl)-lH-indole- l-carboxylate (3.0 g, 9.9 mmol) was added to anhydrous Tl IF (29 mL) and cooled to -78 °C A 0.5M solution of potassium hexuinethyldisilazane (20 mL, 9.9 mmol) was added slowly such that the internal temperature stayed below -60 °C Stirring was continued for 1 h at -78 °C. Methyl iodide (727 μί, 1 1.7 mmol) was added to the mixture. The mixture was stirred for 30 minutes at room temperature. The mixture was quenched with sat. aq. ammonium chloride and partitioned between water and dichloromelhane. The aqueous phase was extracted with dichloromethane and the combined organic phases were dried over Na2S04 and evaporated under reduced pressure. The residue was purified by column- chromatography on silica gel (ethylacetate hexane = 1/9) to give tert-bulyl 2-(l-ethoxy- l-oxopropan-2-yl)- lH-indole- l -carboxylate (2.8 g, 88%).
[00814] Ethyl 2-(lH-indol-2-yl)propanoate
[00815] rm-Butyl 2-( 1 -ethoxy- 1 -oxopropan-2-yl)- 1 H-indole- ! -carboxylate (2.77 g, 8.74 mmol) was dissolved in dichloromelhane (25 mL) before TFA (9.8 mL) was added. The mixture was stirred for 1.5 h at room temperature. The mixture was evaporated to dryness, taken up in dichloromelhane and washed with sat. aq. sodium bicarbonate, water, and brine. The product was purified by column chromatography on silica gel (0-20% EtOAc in hexane) to give ethyl 2-(l H-indol-2-yl)propanoate (0.92 g, 50%).
[00816] Ethyl 2-(5-n ro-lH-indol-2-yl)propanoate
[00817] Ethyl 2-( U I-indol-2-yl)propanoate (0.91 g, 4.2 mmol) was dissolved in concentrated sulfuric acid (3.9 mL) and cooled to -10 °C (sall/ice-mixlure). A solution of sodium nitrate (0.36 g, 4.2 mmol) in concentrated sulfuric acid (7.8 mL) was added dropwise over 35 min. Stirring was continued for another 30 min at -10 °C. The mixture was poured into ice and the product was extracted with ethyl acetate. The combined organic phases were washed with a small amount of sat. aq. sodium bicarbonate. The product was purified by column chromatography on silica gel (5-30% EtOAc in hexane) to give ethyl 2-(5-niln lH-indol-2-yl)propanoalc (0.34 g, 31 ).
[00818] Ethyl 2-{5-amino-lH-indol-2-yl)propanoate
[00819] To a solution of ethyl 2-(5-nitro- lll-indol-2-yl)propanoate (0.10 g, 0.38 mmol) in ethanol (4 mL) was added tin chloride dihydrate (0.431 g, 1.91 mmol). The mixture was heated in the microwave at 120 °C for 1 h. The mixture was diluted with ethyl acetate before water and saturated aqueous NallCOj were added. The reaction mixture was filtered through a plug of cclite using ethyl acetate. rl"hc organic layer was separated from the aqueous layer. rl¾e organic layer was dried over N 2S0 , filtered and evaporated under reduced pressure to give ethyl 2-(5-amino- l H-indol-2-yl)propanoale (0.088 g, 99%).
[00820] Ethyl 2-(5-(l-(benzo[d][l^]dioxol-5-yl)cyclopropanecarboxamido)-lH-indol-2-yl)propanoate [00821 ] To a solution of l -(benzo[d|| 1 ,3]dioxol-5-yl)cyclopropanecarboxylic acid (0.079 g, 0.384 mmol) in acetonitrile ( 1.5 niL) were added HBTU (0. 146 g, 0.384 mmol) and Et3N ( 160 μ1_, 1.15 mmol) at room temperature. The mixture was allowed to stir at room temperature for 10 min before a solution of ethyl 2-(5-amino- l H-indol-2-yl)propanoate (0.089 g, 0.384 mmol) in acetonitrile (2.16 niL) was added. After addition, the reaction mixture was stirred at room temperature for 2 h. The solvent was evaporated under reduced pressure and the residue was dissolved in dichloromethanc. The organic layer was washed with 1 N HC1 (1 3 mL) and then saturated aqueous NaHCC^ ( 1 3 mL). The organic layer was dried over Na2S04, filtered and evaporated under reduced pressure. The crude material was purified by column chromatography on silica gel (ethyl acetate hexane = 1/1) to give ethyl 2-(5-(l-(t>enzo[d][ l,3]dioxol-5-yl)cyclopropanecarboxamido)- l H-indol-2-yDpropanoale (0.081 g, 50%). Ή NMR (400 MHz, CDC13) δ 8.5 1 (s, 1 I I), 7.67 (s, 1H), 7.23-7.19 (m, 2H), 7.04-7.01 (m, 3H), 6.89 (d, J = 0.0 Hz, 1 H), 6.28 (s, lH), 6.06 (s, 2H), 4.25-4.17 (m, 211), 3.91 (q, J = 7.2 Hz, 1 H), 1.72-1.70 (m, 2H), 1.61 (s, 2H), 1.29 (l, J = 7.1 Hz, 411), 1. 13-1.1 1 (m, 211).
[00822] Example 71: terf-Butyl 2-(5-(Mbenzo[d][l,3]dioxol-5-yJ)cyclopropanecarbox-amido)-lH-indol-2-yl)-2-melhylpropylcarbamate
[00823] 2-MethyL.2-(5-nitro-lH-indol-2-yl)propanoic acid
[00824] Ethyl 2-mcthy]-2-(5-nitro- lH-indol-2-yl)propanoate (4.60 g, 16.7 mmol) was dissolved in THF/watcr (2: 1 , 30 mL). LiOH H20 (1.40 g, 33.3 mmol) was added and the mixture was stirred at. 50 °C for 3 h. The mixture was made acidic by the careful addition of 3N HCl. The product was extracted with ethylacetate and the combined organic phases were washed with brine and dried over magnesium sulfate to give 2-methyl-2-(5-nitro- l l l-indol-2-yOpropanoic acid (4.15 g, 99%).
[00825] 2-Methyl-2-(5-nitro-lH-indo]-2-yl)propanamide
[00826] 2-Methyl-2-(5-nitro- lH-indol-2-yi)-propanoic acid (4.12 g, 16.6 mmol) was dissolved in acetonitrile (80 mL). EDC (3.80 g, 0.020 mmol), HOlit (2.70 g, 0.020 mmol), L¾N (6.9 mL, 0.050 mmol) and ammonium chloride ( 1.34 g, 0.025 mmol) were added and the mixture was stirred overnight at room temperature. Water was added and the mixture was extracted with ethylacetate. Combined organic phases were washed with brine, dried over magnesium sulfate and dried to give 2-methyl-2-(5-nitro-lH-indol-2-yl)propanamide (4.3 g, 99%).
[00827] 2-Met yl-2-(5-nitiO-lH-indol-2-yI)propan.l-amine
[00828] 2- ethy]-2-(5-nitro- lH-indol-2-yl)propananiide (200 mg, 0.81 mmol) was suspended in THF (5 ml) and cooled to 0 °C. Borane-THF complex solution (1.0 M, 2.4 mL, 2.4 mmol) was added slowly and the mixture was allowed to stir overnight at room temperature. The mixture was cooled to 0 °C and carefully acidified with 3 N HCl. THF was evaporated off, water was added and the mixture was washed with ethylacetate. The aqueous layer was made alkaline with 50% NaOH and the mixture was extracted with ethylacetate. 'ITie combined organic layers were dried over magnesium sulfate, filtered and evaporated to give 2-methyl-2-(5-nitro- l H-indol-2-yl)propan- l-amine (82 mg, 43%).
[00829] tert-Butyl 2-methyl-2-(5-nitro-lH-indol-2-yl)propylcarbamale
[00830] 2-Methyl-2-(5-nitro- 1 H-indol-2-yl)propan- 1 -amine ( 137 mg, 0.587 mmol) was dissolved in THP (5 niL) and cooled to 0 °C Et3N (82 jiL, 0.59 mmol) and di-wrt-butyl dicarbonate (129 mg, 0.587 mmol) were added and the mixture was stirred at room temperature overnight. Water was added and the mixture was extracted with ethylacetaie. The residue was purified by silica gel chromatography ( 10-40% ethylacetate in hexane) to give ferf-butyl 2-methyl-2-(5-niLro- lll-indol-2-yl)propylcarbamate (131 mg, 67%). [00831 ] tort-Buty [ 2-(5-amino-lH-indol-2-yl)-2-methylpropylcarbamate
[00832] To a solution of tert-butyl 2-methyl-2-(5-nitro- lH-indol-2-yl)propy]carbamate (80 mg, 0.24 mmol) in THF (9 mL) and water (2 niL) was added ammonium formate (60 mg, 0.96 mmol) followed by 10% Pd/C (50 mg). The mixture was stirred at room temperature for 45 minutes. Pd/C was filtered off and the organic solvent was removed by evaporation. The remaining aqueous phase was extracted with dichloromethane. The combined organic phases were dried over magnesium sulfate and evaporated to give rerr-butyl 2-(5-anvino- l H-indol-2-yl)-2-methylpropylcarbaniate (58 mg, 80%).
[00833] tert-Butyl 2-(5-(l-(benzo[d][l ,3]dioxo.-5-yl)cyclopropanecarboxamido)-lH-indol-2-yl)-2-methylpropylcarbamate
[00834] f«?rt-Butyl 2-(5-amino-lII-indoU2-yl)-2-methy1propylcarbamate (58 mg, 0.19 mmol), 1 -(benzoj d][ l,3]dioxol-6-yl)cyclopropanecarboxylic acid (47 mg , 0.23 mmol), EDC (45 mg, 0.23 mmol), HOBt (31 mg, 0.23 mmol) and I¾N (80 μΐ ,, 0.57 mmol) were dissolved in DMF (4 mL) and stirred overnight at room temperature. The mixture was diluted with water and extracted with ethylacetate. The combined organic phases were dried over magnesium sulfate and evaporated to dryness. The residue was purified by silica gel chromatography ( 10-30% ethylacetate in hexane) to give fer/-butyl 2-(5-( l - (benz()|d][l ,3jdioxol-5-yl)cycl()propanecarboxainido)- l l [-indo]-2-yl)-2-mcLhy]pr()pyl-carbamate (88 mg, 94%). Ή NMR (400 MHz, CDC13) 5 8.32 (s, 1 H), 7.62 (d, J = 1.5 Hz, 1H), 7.18 - 7.16 (m, 2H), 7.02 - 6.94 (m, 3H), 6.85 (d, J = 7.8 Hz, 1 H), 6.1 (d, J = 1.5 Hz, l H), 6.02 (s, 2H), 4.54 (m, ! H), 3.33 (d, J = 6.2 Hz, 2H), 1.68 (dd, J = 3.7, 6.8 Hz, 2H), 1.36 (s, 911), 1.35 (s, 6H), 1.09 (dd, J = 3.7, 6.8 Hz, 21-1).
Example 72: (/i)-/V-(2-^ri-Butyl-l-(2>3-dihydroxypropyl)-lH-indol-5-yl)-l- (2,2'difluorobenzo[d][l,3]diox l-5-yl)cyclopropanecarboxamide
[00836] (/?)-2-te/t-Butyl-l-((2,2-dimethyl-lr3.dioxolan-4-yt)methyl)-5-nitro-lH-indole
[00837] To a stirred solution of (5 (2,2-dimc yl-l ,3-dioxolan-4-yl)methy1 4-methylbenzenesulfonate ( 1.58 g, 5.50 mmol) in anhydrous DMF ( 10 inL) under nitrogen gas was added 2-«rt-butyI-5-nitro- l H-indole ( 1.00 g, 4.58 mmol) followed by Cs2C03 (2.99 g, 9.16 mol). The mixture was stirred and heated at 80 °C under nitrogen gas. After 20 hours, 50% conversion was observed by LCMS. The reaction mixture was re-treated with CS2CO3 (2.99 g, 9.16 mol) and (S)-(2,2-dimethyl- l ,3-dioxolan-4-yl)methyl 4-methylbenzenesulfonate (1.58 g, 5.50 mmol) and heated at 80 "C for 24 hours. The reaction mixture was cooled to room temperature. 'Ihe solids were filtered and washed with ethyl acetate and hexane ( 1 : 1).
The layers were separated and the organic layer was washed with water (2 10 mL) and brine (2 x 10 mL). The organic layer was dried over a2S04, filtered and evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (diehloromethane hexane = 1 .5/1 ) to give (R)-2-tert-buty\- 1 -((2, 2 -dimethyl- 1 ,3-dioxo1an-4-yl)melhyl)-5-nitro-lH-indolc ( 1.0 g, 66%). ' H NMR (400 M Hz, CDC13) δ 8.48 (d, J = 2.2 Hz, 1H), 8.08 (dd, J = 2.2, 9. 1 Hz, 1H), 7.49 (d, J = 9. 1 Hz, I H), 6.00 (s, 1 H), 4.52-4.45 (m, 3H), 4.12 (dd, J = 6.0, 8.6 Hz, 1 H), 3.78 (dd, J = 6.0, 8.6 Hz, 1 H), 1 .53 (s, 3H), 1 . 1 (s, 9H), 1.33 (s, 3 H).
[00838] (Wi-Z-iert-Butyl-l-itZ^-dimet yl-l^-dioxolan^-y^methyl-lH-indol-S-ainine
[00839] To a stirred solution of (R)-2-tert-buty\- l-((2,2-dimethyl- l,3-dioxolan-4-yl)methyl)-5-nitro- 1 I l-indok ( 1.0 g, 3.0 nimol) in cthanol (20 mL) and water (5 mL) was added ammonium fomiale (0.76 g, 12 nimol) followed by slow addilion of 10 % palladium on carbon (0.4 g). 'I'he mixture was stirred at room temperature for 1 h. The reaction mixture was filtered through a plug of celitc and rinsed with ethyl acetate. I'he filtrate was evaporated under reduced pressure and the crude product was dissolved in ethyl acetate. The organic layer was washed with water (2 5 mL) and brine (2 5 mL). The organic layer was dried over Na2S0 , fi ltered and evaporated under reduced pressure to give (/?)-2-ter/-butyl- l -((2,2-dimelhyl-l ,3-dioxolan-4-yl)methyl-l /-indol-5-aminc (0.89 g, 98%). Ή NMR (400 MHz, CDC13) 6 7.04 (d, J = 4 Hz, 1 H), 6.70 (d, J = 2.2 Hz, 1 H), 6.48 (dd, ./ = 2.2, 8.6 Hz, 1H), 6.05 (s, 1 H,), 4.38-4. 1 (m, 2H), 4.21 (dd, J = 7.5, 16.5 Hz, II I), 3.87 (dd, J = 6.0, 8.6 Hz, 1 H), 3.66 (dd, J = 6.0, 8.6 Hz, 1 H), 3.33 (br s, 2H), 1 .40 (s, 3H), 1 .34 (s, 9H), 1 .25 (s, 3H). ooY
[00840] N-((fi)-2-½rt-Butyl- l-((2,2-dimethyl-l,3-dioxolan-4-y])methyl)-lH-indol-5-yl)-l-(2,2-difluorobenzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamide
[00841] To 1 -(benzol d][ 1 , 3]dioxol-5-yl)cyclopropanecarboxylie acid (0.73 g, 3.0 mmol) was added Ihionyl chloride (660 uL, 9.0 mmol) and DMF (20 μΐ^) al room temperature. The mixture was stirred for 30 minutes before the excess thionyl chloride was evaporated under reduced pressure. To the resulting acid chloride, dichloromethane (6.0 mL) and 1¾N (2. 1 mL, 15 mmol) were added. A solution of (^^-ieri-butyl-l-^^-dimcthyl- l ^-dioxolan^-ylimethyl- l H-indol-5-amine (3.0 mmol) in dichloromethane (3.0 mL) was added to the cooled acid chloride solution. Alter addition, the reaction mixture was stirred at room temperature for 45 minutes. The reaction mixture was filtered and the filtrate was evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (ethyl acctatc/hcxane = 3/7) to give Ν-((ϋ)-2-½rr-bulyl- l-((2,2-dimelhyl- 1 ,3-dioxolan-4-yl)methyl)- 1 H-indol-5-yl)- 1 -(2,2-difluorobenzo[d][ l ,3]dioxol-5-yl)cyclopropanecarboxamide ( 1.33 g, 84%). Ή NMR (400 MHz, CDClj) δ 7.48 (d, J = 2 Hz, 111,), 7.3 1 (dd, J = 2, 8 Hz, 1 H), 7.27 (dd, J = 2, 8 Hz, 1H), 7.23 (d, J = 8 Hz, 1 H), 7.14 (d, J = 8 Hz, 1 H), 7.02 (dd, J = 2, 8 Hz, 1H), 6.92 (br s, lH), 6.22 (s, 1 H), 4.38-4.05 (m, 3H), 3.91 (dd, J = 5, 8 Hz, 1 H), 3.75 (dd, J = 5, 8 Hz, 1 H), 2.33 (q, J = 8 Hz, 2H), 1.42 (s, 3H), 1.37 (s, 9H), 1.22 (s, 3H), 1.10 (q, J = 8 Hz, 2H).
[00842] N-((«)-2-/ert-Butyl-l-((2,3-dihydroxypropyI)-l -indol-5-yl)-l-(2,2-difluorobenzo-[d][l,3]dioxol-5-yl)cyclopropanecarboxamide
[00843] To a stirred solution of N-(2-/er/-butyl-l-((2,2-dimelhyl- 1 ,3-dioxolan-4-yl)niethyl)-l H-indol-5-yl)-l-(2,2-difluorobenzo[dJ[ l ,3]dioxol-5-yl)cyclopropanecarboxamide (1.28 g, 2.43 mmol) in methanol (34 ml.,) and water (3.7 ml.,) was added para-toluenesulfonic acid-hydrate ( 1.87 g, 9.83 mmol). The reaction mixture was stirred and heated at 80 °C for 25 minutes. The solvent was evaporated under reduced pressure. The crude product was dissolved in ethyl acetate. The organic layer was washed with saturated aqueous NaHCC>3 (2 x 10 mL) and brine (2 x 10 mL). The organic layer was dried over a2SC>4, filtered and evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (ethyl acclate hexanc = 13/7) to give /V-((^)-2-ierr-butyl-l -((2,3-dihydroxypropyl)- l / -indol-5-yl)- l -(2,2-difiuorobenzo[d][ 1 ,3]dioxol-5-yl)cyclopropanecarboxamide (0.96 g, 81 %). Ή NMR (400 MHz, CD(¾) 67.50 (d, J = 2 Hz, 1H), 7.31 (dd, J = 2, 8 Hz, 1 H), 7.27 (dd, J = 2, 8 Hz, 1H), 7.23 (d, J = 8 Hz, H I), 7.14 (d, J - 8 Hz, H I), 7.02 (br s, I I I,), 6.96 (dd, J = 2, 8 Hz, 1H), 6.23 (s, III), 4.35 (dd, J = 8, 15 Hz, 1 H), 4.26 (dd, J = 4, 15 Hz, I H,), 4.02-3.95 (m, 1 H), 3.60 (dd, J = 4, 1 1 Hz, 1H), 3.50 (dd, J = 5, Π Hz, 1 H), 1.75 (q, J = 8 Hz, 3H), 1.43 (s, 911), 1.14 (q, J = 8 Hz, 3H).
[00844] Example 73: 3-(2-tert-Butyl-5-(1 -(2,2-difluorobenzo[d][ l,3]dioxol-5-yl)cyclopropanecarboxamido)-lH-indol-l-yl)-2-hydroxypropanoic acid
[00845] 3-(2-i
[00846] To a solution of /V-(2-½ri-butyl- 1 -(2,3-dihydroxypropy])- 1 H-indol-5-yl)- 1 -(2,2-difluorobenzo[d][ l ,3]dioxol-5-yl)cyclopropane-carboxamide (97 mg, 0.20 mmol) in DMSO (1 mL) was added Dess-Martin pcriodinane (130 mg, 0.30 mmol). The mixture was stirred at room temperature for 3 h. The solid was filtered off and washed with BtOAc. The filtrate was partitioned between BtOAc and water. The aqueous layer was extracted with EtOAc twice and the combined organic layers were washed with brine and dried over MgSC^. After the removal of solvent, the residue was purified by preparative TLC to yield 3-(2-ier/-bulyl-5-( 1 -(2,2-difluorobenzo[d) [ 1 ,3]dioxol-5-yl)cyclopropanecarboxamido)- 1 H-indol- l-yl)-2-oxopropanoic acid that was used without further purification.
[00847] 3-(2-it?rt-ButyI-5-(l-(2,2-difIuorobenzo[d][l^]dioxol-5-yI)cyclopropanecarbox-amido)-lH-indol-l-yl)-2-hydroxypropanoic acid
[00848] To a solution of 3-(2-/t?rt-bulyl-5-( 1 -(2,2-difluorobenzo[d][ 1 ,3]dioxol-5-yl)cyclopropanecarboxamido)- l H-indol- l -yl)-2-oxopropanoic acid (50 mg, 0. 10 minol) in MeOI I ( 1 niL) was added NaBH4 ( 19 mg, 0.50 mmol) at 0 °C. The mixture was stirred at room temperature for 15 min. The resulting mixture was partitioned between EtOAc and water. 'ITic aqueous layer was extracted with EtOAc twice and the combined organic layers were washed with brine and dried over anhydrous MgS04. After the removal of the solvent, the residue was taken up in DMSO and purified by preparative LC MS to give 3-(2-tert-bulyl-5-{ l-(2,2-difluorobenzo[d]| 1 ,3]dioxol-5-yl)cyclopropanecarboxaniido)-lH-indol- l -yl)-2-hydroxypropanoic acid. ]H NMR (400 MHz, COCI3) δ 7.36 (s), 7.27-7.23 (m, 2H), 7.15-7.1 1 (m, 2H), 6.94 (d, J = 8.5 Hz, I II), 6.23 (s, 1 H), 4.71 (s, 311), 4.59 (q, J = 10.3 Hz, 1H), 4.40-4.33 (m, 2H), 1.70 (d, J = 1 .9 Hz, 2H), 1.15 (q, J = 4.0 Hz, 2H). I3C NMR (400 MHz, CDClj) δ 173.6, 173.1 , 150.7, 144.1 , 143.6, 136.2, 135.4, 134.3, 131.7, 129.2, 129.0, 127.6, 126.7, 1 16.6, 1 14.2, 1 12.4, 1 10.4, 1 10. 1 , 99.7, 70.3, 48.5, 32.6, 30.9, 30.7, 16.8. MS (ESI) m/e (M-i-H+) 501.2.
[00849] Example 74: (/i)-N-(2-½rt-Butyl-l-(2,3-dihydroxypropyl)-lH-indol.5-yl)-l-(2,2-dideuteriumbenzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamide
[00850] Methyl l -(3,4-dihydroxyphenyl)cyclopn>panecarboxylate
[00851] To a solution of 1 -(3,4-dihydroxyphenyl)cyclopropanecarboxylic acid ( 190 mg, 1.0 mmol) in MeOH (3 mL) was added 4-methylbenzenesulfonic acid (19 mg, 0.10 mmol). The mixture was heated al 80°C overnight. The reaction mixture was concentrated in vacuo and partitioned between EtOAc and water. The aqueous layer was extracted with EtOAc twice and the combined organic layers were washed with sal. NaHCC and brine and dried over MgS04. After die removal of solvent, the residue was dried in vacuo to yield melhyl I.-(3,4-dihydroxyphenyl)cyclopropanecarboxylate ( 190 mg, 91 %) thai was used without further purification. Ί I NMR (400 MHz, DMSO-d6) 8 6.76-6.71 (m, 2H), 6.66 (d, J = 7.9 Hz, 1 H), 3.56 (s, 3H), 1.50 (q, J = 3.6 Hz, 2H), 1.08 (q, J = 3.6 Hz, 211).
[00852] Methyl 1 -(2,2-dideuteriumbenzo[d][1 ,3]dioxol-5-yl)cyclopropanecarboxylate
[00853] To a solution of methyl l -(3,4-dihydroxyphenyl)cyciopropanecarboxylate (21 mg, 0.10 mmol) and CD2Brz (35 mg, 0.20 mmol) in DMF (0.5 mL) was added Cs2C03 (19 mg, 0.10 mmol). The mixture was heated at 120°C for 30 min. The reaction mixture was partitioned between EtOAc and water. The aqueous layer was extracted with EtOAc twice and the combined organic layers were washed with IN NaOH and brine before being dried over MgS04. After the removal of solvent, the residue was dried in vacuo to yield methyl 1-(2,2-dideuteriumbenzo[d]| l ,3]dioxoL5-y])cyclopropanecarboxylate (22 mg) that was used without further purification. JH NMR (400 MHz, CDC13) δ 6.76-6.71 (m, 2H), 6.66 (d, J = 7.9 Hz, III), 3.56 (s, 311), 1.50 (q, J = 3.6 Hz, 211), 1.08 (q, J = 3.6 Hz, 211).
[00854] 1- (2,2 - Dideuter iu mbenzo [d] [ 1 ,3]dioxol-5-y l)cyclopropanecar boxy lie acid
[00855] To a solution of methyl l -(2,2-dideuteriumbenzo[d][ l ,3 ]dioxol-5-yl)cyclopropanecarboxylale (22 mg, 0.10 mmol) in THE (0.5 mL) was added NaOH ( I N, 0.25 mL, 0.25 mmol). 1 he mixture was heated at 80°C for 2 h. rlhe reaction mixture was partitioned between EtOAc and IN NaOH. The aqueous layer was extracted with EtOAc twice, neutralized with IN HO and extracted with EtOAc twice. The combined organic layers were washed with brine and dried over MgSO,). After the removal of solvent, the residue was dried in vacuo lo yield l -(2,2-dideuteriumbenzold][ l ,3 |dioxol-5-yl)cyclopropaneearboxylic acid (21 mg) that was used without further purification.
[00856] («)-N-(2-^rt-Butyl- l-((2,2-dimethy[-l,3-dioxolan-4-yl)methyl)-lH-indol.5-yl)-l -(2,2-dideuteriumbenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamide
[00857] To a solution of 1 -(2,2-dideuteriumbcnzo[d][ 1 ,3]dioxol-5-yOcyclopropanccarboxylic acid (21 mg, 0.10 inmol), (/i)-2-/er/-bulyl-l -((2,2-dimcthyl- l ,3-dioxolan-4-yl)methyl)- I H-indol-5-aniine (30 nig, 0.10 mmol), HATU (42 nig, 0.11 mol) in DMF ( 1 mL) was added triethylamine (0.030 mL, 0.22 mmol). The mixture was heated at room temperature for 5 min. The reaction mixture was partitioned between EtOAc and water. The aqueous layer was extracted with EtOAc twice and the combined organic layers were washed with I N NaOI l, I N I-IC1, and brine before being dried over gSO,). After the removal of solvent, the residue was purified by column chromatography (20-40% ethyl acelate/hexane) lo yield ( ?)- V-(2-/eri-butyl-l -((2,2-dinielhyl- 1 ,3-dioxolan-4-y])methyl)- l H-indol-5-yl)-l-(2,2-dideuteriunibenzo[d][ l ,31dioxol-5-yl)cyclopropanecarboxamide (24 mg, 49% from methyl l-(3,4-dihydroxyphenyl)cyclopropanecarboxylale). MS (ESI) m/e (M+H+) 493.5.
[00858] (i?)-N-{2-½rt-Buty]-l-(2,3-dihydroxypropyl)-lH-indol-5-yl)- l.(2,2-dideuterium-benzo[d][l,3]dioxo]-5-y])cyclopropanecarboxamide
[00859] To a solution of ( i)-N-(2^eri-butyl- l-((2,2-diniethy] ,3-dioxo]an-4-yl)melhyl)-lH-indol-5-yl)- l -(2,2-dideuterium-benzo|d] [ l,3]dioxol-5-yl)cyclopropanecarboxaniide (24 mg, 0.050 mmol), in methanol (0.5 mL) and water (0.05 mL) was added 4-methylbenzenesulfonic acid (2.0 mg, 0.010 mmol). The mixture was heated at 80°C for 30 min. The reaction mixture was partitioned between EtOAc and water. The aqueous layer was extracted with EtOAc twice and the combined organic layers were washed with sat. aHC03 unci brine before being dried over MgSC . After the removal of solvent, the residue was purified by preparative HPLC to yield ( i)-Af-(2-ii;ri-butyl- l-((2,2-dimethyl- l ,3-dioxolan-4-y methyl)- 1 H-indol-5-yl)- 1 -(2,2-dideuteriumbenzo[dJ| 1 ,3]dioxol-5-yl)cyclopropanecarboxamide (12 mg, 52%). ]I I NMR (400 MHz, CDC13) δ 7.44 (d, J = 2.0 Hz, 1H), 7.14 (dd, J = 22.8, 14.0 Hz, 2H), 6.95-6.89 (m, 2H), 6.78 (d, J = 7.8 Hz, 1 H), 6.14 (s, 1 H), 4.28 (dd, J = 15. 1, 8.3 Hz, 11-1), 4.19 (dd, J = 15.1 , .5 Hz, 1H), 4.05 (q, J = 7. 1 Hz, 1 H), 3.55 (dd, J = 1 1 .3, 4.0 Hz, 1 H), 3.45 (dd, J = 1 1.3, 5.4 Hz, 1 H), 1 .60 (q, J = 3.5 Hz, 2H), 1.35 (s, 9H), 1.02 (q, J = 3.5 Hz, 2H). 13C NMR (400 MHz, C C13) δ 171.4, 149.3, 147.1 , 146.5, 134.8, 132.3, 129.2, 126.5, 123.6, 1 14.3, 1 1 1.4, 1 10.4, 109.0, 107.8, 98.5, 70.4, 63.1 , 46.6, 31.6, 30.0, 29.8, 15.3. MS (ESI) m e (M+H+) 453.5.
[00860] It is further noted that the mono-deuterated analogue for this compound can be synthesized by substitution the reagent CHDBR2 for CD2BR2 and following the procedures described in example 74. Furthermore, mono-deuterated analogues of other compounds of the present invention can be synthesized by substituting the reagent CHDBR2 for CD2BR2 and following the steps described herein.
[00861] Example 75: 4-(5-(l-(Benzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamido)-lH-indol-2-yl)-4-methylpentanoic acid
[00862] l-(Benzo[d][1 ]dioxol-5-yl).^-(2-(4-cyano-2-methyIbutan-2-yl)-lH-indol-5-yDcyclopropanecarboxamide
[00863] To l-(benzo|d][ l,3Jdioxol-5-yl)cyclopropanecarboxylic acid (0.068 g, 0.33 mmol) was added thionyl chloride (72 μί, 0.99 mmol) and DMF (20 μί) at room temperature. The mixture was stirred for 30 minutes before the excess thionyl chloride was evaporated under reduced pressure. To the resulting acid chloride, dichloroniethane (0.5 raL) and Et3N (230 μΐ,, 1 .7 nimol) were added. Λ solution of 4-(5-amino- l H-indol-2-yI)-4-methylpentanenitrile (0.33 mniol) in dichloromethane (0.5 mL) was added to the acid chloride solution and the mixture was stirred at room temperature for 1 .5 h. The resulting mixture was diluted with dichloromethane and washed with 1 N I IC1 (2 x 2 mL), saturated aqueous NallCO^ (2 x 2 mL) and brine (2 x 2 mL). The organic layer was dried over anhydrous Na2S04 and evaporated under reduced pressure to give l -(benzo[dJ[l ,3 |dioxol-5-yl)-iV-(2-(4-cyano-2-methylbutan-2-yl)- l H-indol-5-yl)cyclopropanecarboxamide.
[00864] 4-(5-(l-(Benzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamido)-lH-indol-2-yl)-4-methylpentanoic acid
[00865] A mixture of l-(bcnzo[d][ 1 ,3]dioxol-5-yl)-N-(2-(4-cyano-2-mcthylbutan-2-yl)- 1 H-indol-5-yl)cyclopropanecarboxamide (0.060 g, 0. 15 inmol) and KOH (0.08 1 g, 1.5 mniol) in 50% EtOH/water (2 mL) was heated in the microwave at 100 °C for 1 h. The solvent was evaporated under reduced pressure. The crude product was dissolved in DMSO (1 mL), filtered, and purified by reverse phase preparative HFLC to give 4-(5-( l -(benzo|d) L l ,3]dioxol-5-yl)cyclopropanecarboxarnido)- l H-indol-2-yl)-4-inethylpentanoic acid. Ή NMR (400 MHz, DMSO-d6) δ 1 1 .98 (s, 1 H), 10.79 (s, 1H), 8.44 (s, 1H), 7.56 (s, 1 H), 7.15 (d, J = 8.6 Hz, 1 H), 7.03-6.90 (m, 4H), 6.05 (s, 111), 6.02 (s, 2H), 1.97- 1 .87 (m, 4H), 1.41 - 1.38 (m, 211), 1.30 (s, 6H), 1 .04- 1.02 (m, 2H).
[00866] Example 76: l-(Benzo[d][l^]dioxol-5-yl)-N-(2-(l-hydroxypropan-2-yl)-l H-indol-5-yl)cyclopropaiiecarboxamide
[00867] 2-(5-Nitro-lH-indol-2-yl)propi
[00868] To a cooled solution of UA1H4 (1 .0 M in THF, 1.2 ml.-, 1.2 mmol) in THF (5.3 mL) at 0 °C was added a solution of ethyl 2-(5-nilro- l I I-indoL2-yl)propanoate (0.20 g, 0.76 mmol) in THF (3.66 mL) dropwise. After addition, the mixture was allowed to warm up to room temperature and was stirred at room temperature for 3 h. The mixture was cooled to 0 °C. Water (2 mL) was slowly added followed by careful addition of 15% NaOH (2 mL) and water (4 mL). The mixture was stirred at room temperature for 0.5 h and was then filtered through a short plug of celite using ethyl acetate. The organic layer was separated from the aqueous layer, dried over Na2S04, filtered and evaporated under reduced pressure. ITie residue was purified by column chromatography on silica gel (ethyl acetatc hcxane = 1/ 1 ) to give 2-(5-nitro-l H-indoI-2-yI)propan- I -oI (0.14 g, 81 %).
[00869] 2-(5-Amino-lH-indol-2-yl)propan-l-oJ
[00870] To a solution of 2-(5-nilro-l H-indol-2-yl)propan-l -ol (0. 13 g, 0.60 mmol) in cthanol (5 mL) was added tin chloride dihydrate (0.67 g, 3.0 mmol). The mixture was heated in the microwave at 120 °C for 1 h. The mixture was diluted with ethyl acetate before water and saturated aqueous NaHCOj were added. The reaction mixture was filtered through a plug of celite using ethyl acetate. The organic layer was separated from the aqueous layer, dried over Na2S04, filtered and evaporated under reduced pressure to give 2-(5-amino- 1 H-indol-2-yl)propan- l -ol (0.093 g, 82%).
[00871] l-(Benzo[d][l,3]dioxo]-5-yl)-N-(2-(l-hydroxypropan-2-yl)-lH-indol-5-yl)cyclopropanecarboxamide
[00872] To a solution of l-(benzo[d]| 1 ,3]dioxol-5-yl)cyclopropanecarboxylic acid (0. 10 g, 0.49 mmol) in acetonilrile (2.0 mL) were added H TU (0.185 g, 0.49 mmol) and 1¾N (205 μί, 1.47 mmol) al room temperature. The mixture was allowed to stir at room temperature for 10 minutes before a slurry of 2-(5-amino- lI-I-indol-2-yl)propan- l -ol (0.093 g, 0.49 mmol) in aeetonitrile (2.7 mL) was added. After addition, the reaction mixture was stirred at room temperature for 5.5 h. The solvent was evaporated under reduced pressure and the residue was dissolved in dichloromethane. The organic layer was washed with I N HC1 (1 x 3 mL) and saturated aqueous NaHCC^ ( 1 x 3 mL). The organic layer was dried over Na2S04, filtered and evaporated under reduced pressure. The crude material was purified by column chromatography on silica gel (ethyl acetatehexane = 13/7) to give l-(benzo[dJ[l,3]dioxol-5-yl)-N-(2-(l-hydroxypr pan-2-yl)-lH-indol-5-yl)cyclopropanecarboxamide (0.095 g, 51%). Ή NMR (400 MHz, DMSO-d6) 610.74 (s, III), 8.38 (s, III), 7.55 (s, III), 7.14 (d, J = 8.6 Hz, III), 7.02-6.90 (m, 411), 6.06 (s, 1H), , 6.02 (s, 2H), 4.76 (t, J = 5.3 Hz, 1H), 3.68-3.63 (m, 111), 3.50-3.44(m, 111), 2.99-2.90 (m, 111), 1.41-1.38 (m, 2H), 1.26 (d, J = 7.0 Hz, 3H), 1.05-1.02 (m,2H).
[00873] Example 77: 1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-1 -indol-5-yl)-N-methylcyclopropanecarboxamide
[00874] l-(Benzo[<][l^]dioxol-5-yl)-N-(2-tert-butyl-l/ -indol-5-yl)-/V-methylcyclopropanecarboxamide
[00875] 2-ieri-Butyl-/V-melhyl-l«-indol-5-amine (20.2 mg, 0.100 mmol) and 1-(benzo[d][l,3]dioxol-5-yl)cyclopropanecarboxylic acid (20.6 mg, 0.100 mmol) were dissolved in NN-dinieLhylformamide (1 mL) containing irielhylamine (42.1 Ι^, 0.300 mmol) and a magnetic stir bar. <5-(7-Azabenzotriazoi- l-yl)-/V,N,N',V"-tetramethyluronium hexafluorophosphale (42 mg, 0.11 mmol) was added lo the mixture and the resulting solution was allowed to stir for 16 h at 80 °C. The crude producl was then purified by preparative HPLC utilizing a gradient of 0-99% acetonitrilc in water containing 0.05% irifluoroacetic acid to yield 1-(benzo|rf|| ],3|dioxoI-5-yl)-N-(2-ifrt-butyl-lW-indol-5-yl)-Vr-melhylcyclopropanecarboxamide. ESI-MS >n/z calc.390.2, found 391.3 (M+l)+. Retention time of 3.41 minutes.
[00876] Example 78: /V-(2-.t?rf-Butyl-1 -methyl- 1H-indol-5-yl)-l -(benzo[d][l,3]dioxol-6-y 1)- N -meth lc clopropanecarboxamide
[00877] Sodium hydride (0.028 g, 0.70 mmol, 60% by weight dispersion in oil) was slowly added to a stirred solution of N-(2-tcrt-butyl-lH-indol-5-yl)-l-(bcnzo[d][l,3|dioxol-6-yl)cyelopropanecarboxamide (0.250 g, 0.664 mmol) in a mixture of 4.5 mL oi" anhydrous tetrahydrofuran (ΤΗΓ) and 0.5 mL of anhydrous NN-dimethylfornianiide (DMF). The resulting suspension was allowed to stir for 2 minutes and then iodomelhane (0.062 ml.,, 1.0 mmol) was added to the reaction mixture. Two additional aliquots of sodium hydride and iodoniethane were required to consume all of the starting material which was monitored by LC / MS. The crude reaction product was evaporated to dryness, redissolved in a minimum of DMF and purified by preparative LC / MS chromatography to yield the pure product (0.0343 g, 13%) ESI-MS m/z calc. 404.2, found 405.3 (M+ 1 )+. Retention time of 3.65 minutes.
[00878] Example 79: l -(Benzo[d][1,3]dioxol-5-yl)-N-(2-(hydroxymethyl)-l H-indol-5-yl)cyclopropanecarboxamide
[00879] Ethyl 5-(l-(benzo[d]l l ,3|dioxol-5-yl)cyclopropanecarboxamido)- l I I-indole-2-carboxylate (1.18 g, 3.0 mmol) was added lo a solution of LiBH4 (132 mg, 6.0 mmol) in THF ( 10 mL) and water (0.1 mL). The mixture was allowed to stir for I6h at 25 °C before it was quenched with water (10 mL) and slowly made acidic by addition of 1 N HC1. The mixture was extracted with three 50-mL portions of ethyl acetate. The organic extracts were dried over Na2S04 and evaporated to yield 1 -(benzoyl! 1 , 31dioxol-5-yl)-N-(2-(hydroxymeihyl)-l H-indol-5-yl)cyclopropanecarboxamide (770 mg, 73%). Λ small amount was further purified by reverse phase HPLC. ESI-MS m/z calc. 350.4, found 351 .3 (M+l )+; retention time 2.59 minutes.
[00880] Example 80: 5-(l-(Benzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamido)-A'-ifrt-butyl-lH-indole-2-carboxamide
[00881] 5-(l-(Benzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamido)-lH-indoIe-2-carboxylic acid
[00882] Ethyl 5-(l -(benzo[d]| l ,3 |dioxol-5-yl)cyclopropanecarboxamido)- l H-indole-2-carboxylate (392 mg, 1.0 mmol) and LiOH (126 mg, 3 mmol) were dissolved in H20 (5 mL) and 1 ,4-dioxane (3 mL). The mixture was heated in an oil bath at. 100 °C for 24 hours before it was cooled to room temperature. ITie mixture was acidified with I HCI and it was extracted with three 20 mL portions of dichloromethane. The organic extracts were dried over Na2SOj and evaporated to yield 5-( I -(benzotd]| 1 ,3]-dioxol-5-yl)cyclopropanecarboxaniido)- l H-indole-2-carboxylic acid (302 mg, 83%). A small amount was further purified by reverse phase HPLC. ESI-MS in/i calc. 364.1 , found 365.1 (M+ l)+; retention time 2.70 minutes.
[00883] 5-(l-(benzo[d][l,3)dioxol-5-yl)cyclopropanecarboxamido)-/V-iert-butyl-lH-indole-2-carboxaniide
[00884] 5-(l-(Benzo[dl| l ,3 |dioxol-5-yl)cyclopropane-carboxamido)-l H-indolc-2-carbox lic acid (36 mg, 0.10 nimol) and 2-niethylpropan-2-amine (8.8 mg, 0. 12 mmol) were dissolved in V,/V-diniethylformamide (1 .0 mL) containing iriethylamine (28 lL, 0.20 mmol). 0-(7-Azabenzotriazol- L-yl)-jV,/V,/V',N'-tetramethyluronium hexafluorophosphate (46 mg, 0.12 mmol) was added to the mixture and the resulting solution was allowed to stir for 3 hours. The mixture was filtered and puri ied by reverse phase HPLC to yield 5-(l -(benzo[d]| 1 ,3 |dioxol-5-yl)cyclopropanecarboxamido)-N-iert-butyl- LH-indole-2-carboxamide. ESI-MS m/z calc. 419.2, found 420.3 (M+l )+; retention lime 3.12 minutes.
[00885] Example 81: N-(3-Amino-2-fert-butyl-lH-indol-S-yl)-l-(berizo[d][l,3]dioxol-5-yl)cyclopropanecarboxamide
[00886] A solution of 1 -(benzo[d][l ,3]dioxoJ-5-yl)-N-(2-tert-butyl- 1 H-indol-5-yl)cyclopropane carboxamide (50 mg, 0.13 mmol) was dissolved in AcOH (2 mL) and wanned to 45 °C. To the mixture was added a solution of NaN02 (9 mg) in H20 (0.03 mL). The mixture was allowed to stir for 30 min at 45 °C before the precipitate was collected and washed with Et20. This material was used in the next step without further purification. To th crude material, l -(benzold][ l ,3]dioxol-5-yl)-N-(2-tert-butyl-3-nitroso-1 H-indol-5-y cyclopropanecarboxamide, was added AcOH (2 mL) and Zn dust (5 trig). The mixture was allowed to stir for I h al ambient temperature. ElOAc and H2O were added to the mixture. The layers were separated and the organic layer was washed with sat. aq. Nal lCO^, dried over MgS04, and concentrated in vacuo. The residue was taken up in DMF ( 1 mL) and was purified using prep-HPLC. LCMS: m/z 392.3; retention time of 2.18 min.
[00887] Example 82: l-(Benzo[d][l,3]dioxol-5-yl)-N-(2-½rt-butyl-3-(methylsulfonyl). l H-indol-5-yl)cyclopropanecarboxamide
[00888] l-tBenzotdlt ldioxol-S-ylJ-N-il-tert-butyl-a-dnethylsulfonylJ-lH-indol-S-yl)cyclopropanecarboxamide
[00889] To a solution of i-(benzo[d]L l ,3]dioxol-5-yl)-JV-(2-/ert-butyl-l H-indol-5-yl)eyclopropanecarboxamide ( 120 mg, 0.31 mmol) in anhydrous DMF-THF (3.3 mL, 1 :9) was added NaH (60% in mineral oil, 49 mg, 1.2 mmol) at room temperature. After 30 min under N2, the suspension was cooled down to -15 "C and a solution of methanesulfonyl chloride (1.1 eq.) in DMF (0.5 mL) was added dropwise. The reaction mixture was stirred for 30 min at -15 °C then for 6 h at room temperature. Water (0.5 mL) was added at 0 °C, solvent was removed, and the residue was diluted with MeOH, filtrated and purified by preparative HPLC to give l-(Benzo[d]l l ,3]dioxol-5-yl)-N(2-ieri-butyl-3-(methylsullbnyl)-lH-indol-5-yl)cyclopropanecarboxamide. Ή NMR (400 MHz, DMSO) δ 1 1.6 (s, I H), 8.7 (s, 1H), 7.94 (d, J = 1.7 Hz, 1H), 7.38 (d, J =8.7 Hz, 1 H), 7.33 (dd, Jl =1.9 Hz, J2 =8.7 Hz, IH), 7.03 (d, ,/ =1.7 Hz, IH), 6.95 (dd, Jl =1.7 Hz, J2 =8.0 Hz, IH), 6.90 (d, J =8.0 Hz, I H), 6.02 (s, 2H), 3.07 (s, 311), 1.56- 1.40 (m, 911), 1.41 (dd, Jl =4.0 Hz, 72 =6.7 Hz, 2H), 1.03 (dd, Jl =4.0 Hz, J2 =6.7 Hz, 2H). MS (ESI) m/e (M+H+) 455.5.
[00890] Example 83: l-(Benzo[rf][l,3]dioxol-S-yl)-N-(3-phenyl-lH-indol-5-yl)cyclopropane carboxamide OH [00891 ] l -(Benzo[i ][l ,3]dioxol-5-yl)-N-(3-bromo-l -indol-5-y l)c yclopropanecar boxami de
[00892] Freshly recrystallized iV-broinosuccinimdc (0.278 g, 1 .56 mmol) was added portion wise to a solution of I -(benzol l ,3]dioxoL5-yl)-iV-( I W-indol-5-yl)cyclopropanccarboxamide (0.500 g, 1.56 mniol) in MN-dimcthylformamidc (2 mL) over 2 minutes. The reaction mixture was protected from light and was stirred bar for 5 minutes. The resulting green solution was poured into 40 mL of water. The grey precipitate which formed was filtered and washed with water to yield l -(benzo| i/H 1,3 jdioxol-5-yl)-N-(3-bromo- l//-indol-5-yl)cyclopropanecarboxamide (0.564 g, 9 1 %). ES1-MS m/z calc. 398.0, found 399.3 (M+l )+. Retention time of 3.38 minutes. Ή NMR (400 MHz, DMSO-rf6) 1 1.37 (s, 1 H), 8.71 (s, 1H), 7.67 (d, J = 1.8 Hz, I H), 7.50 (d, J = 2.6 Hz, 1 H), 7.29 (d, J = 8.8 Hz, 1 H), 7.22 (dd, J = 2.0, 8.8 Hz, 1 H), 7.02 (d, J = 1.6 Hz, 1 H), 6.96 - 6.88 (m, 211), 6.03 (s, 2H), 1.43 - 1 .40 (m, 2H), 1 .09 - 1.04 (m, 2H).
[00893] l-(Benzo[rf][l,3]dioxol-S-yl)- -(3-phenyl-lH-indol-5-yl)cyctopropanecarboxamide
[00894] Phenyl boronic acid (24.6 mg, 0.204 mniol) was added to a solution of 1 -(benzo[rf]| LSl-dioxol-S-y - -iS-bromo- l iZ-indol-S-y cyclopropanecarboxamidc (39.9 mg, 0.100 mmol) in ethanol ( 1 mL) containing FibreCal 1001 (6 trig) and I M aqueous potassium carbonate (0.260 mL). The reaction mixture was then heated at 130 °C in a microwave reactor for 20 minutes. The crude product was then purified by preparative HPLC utilizing a gradient of 0-99% acetonitrile in water containing 0.05% trifluoroacetic acid to yield 1-(benzo[i/][ 1, Idioxol-S-yD-N-CS-phenyl- 1 /-indol-5-yl)cyclopropane carboxamide. ESI-MS /z calc. 396.2, found 397.3 (M+ l )+. Retendon time of 3.52 minutes. Ή NMR (400 MHz, DMSO-rf6) δ 1 1.27 (d, J = 1 .9 Hz, 1H), 8.66 (s, 1 H), 8.08 (d, J = 1.6 Hz, 1H), 7.65-7.61 (m, 3H), 7.46-7.40 (m, 2H), 7.31 (d, J = 8.7 Hz, I H), 7.25-7. 17 (m, 2H), 7.03 (d, J 6.98-6.87 (m, 2H), 6.02 (s, 2H). 1.43- 1.39 (m, 2H), 1.06- 1.02 (m, 21-1).
[00895] Example 84: l-(Benzotd]tl,3]dioxol-S-yl)-N-(2-tert-butyl-3-cyano-lH-indol-5-yl)cyclopropanecarboxamide
[00896] l -(Benzo[d][l,3]dioxol-5-yl)-N-(2-iert-butyl-3-formyl- lH-indol-5-y l)cyclopr opane-car b xamidc
[00897] P0C13 (12 g, 80 mraol) was added dropwise to DMF (40 mL) held ai -20 °C. After the addition was complete, the reaction mixture was ailowcd to warm to 0 °C and was stirred for 1 h. 1 -(Benzo[d |[ 1 ,3]dioxol-5-yl)-N-(2-i£?«-butyl- 1 H-indol-5-yl)cyclopropanecarboxamide (3.0 g, 8.0 niniol) was added and the mixture was wanned to 25 °C. After stirring for 30 minutes the reaction mixture was poured over ice and stirred for 2 h. The mixture was then heated at 100 °C for 30 min. The mixture was cooled and the solid precipitate was collected and washed with water. The solid was then dissolved in 200 mL dicruoromethane and washed with 200 mL of a saturated aq. NallCOj. The organics were dried over Na2S04 and evaporated to yield l-(benzo[d][ l ,3]dioxol-5-yI)-N-(2-re'ri-butyl-3-formyl-l H-indol-5-yl)cyclopropane-carboxamide (2.0 g, 61 %). ESLMS m/z calc. 404.5, i'ound 405.5 (M+l )+; retention time 3.30 minutes. Ή NM (400 MHz, DMSO-d6) δ 1 1.48 (s, 1 H), 10.39 (s, 1H), 8.72 (s, 1H), 8.21 (s, 1H), 7.35-7.3 1 (m, 2H), 7.04-7.03 (m, 1 H), 6.97-6.90 (m, 2H), 6.03 (s, 2H), 1.53 (s, 9H), 1.42- 1.39 (m, 2li), 1.05- 1.03 (m, 2H).
[00898] ( J-l-iBenMldlll^ldioxol-S- O-N-i -ieri-but l-S-ith drox iminoinieth ])-!!!-indol-5-y))cyclopropanecarboxamide
[00899] To a solution of l-(benzo|d|[l ,3]dioxol-5-yl)-.V-(2-ter/-bulyl-3-lbnnyl-l H-indol-5-yl)cyclopropanecarboxamide (100 mg, 0.25 mmol) in dichloromethane (5 mL) was added hydroxylamine hydrochloride (21 mg, 0.30 mmol). After stirring for 48 h, the mixture was evaporated to dryness and purified by column chromatography (0-100% ethyl acctatehexancs) to yield (Z)-l-(bcnzo|d]| l,3]dioxol-5-yl)-Ar-(2-ieri-bulyl-3-((hydroxyimino)methyl)-UI-indol-5-yl)cyclopropaneearboxaniide (81 mg, 77%). ES1- S mz calc.419.5, found 420.5 (M+l)+; retention time 3.42 minutes. Ή NMR (400 MHz, D SO-d6) δ 10.86 (s, 0.5H), 10.55 (s,0.5H), 8.56-8.50 (m, 211), 8.02 (m, 111), 7.24-7.22 (m, IH), 7.12-7. L0 (m, III), 7.03 (in, IH), 6.96-6.90 (m, 2H), 6.03 (s, 2H), 1.43 (s, 9H), 1.40-1.38 (m, 211), 1.04-1.01 (m, 2H).
[00900] 1-(Benzo[d][1,3]dioxol-5-yl)-N-(2-tert-butyl-3-cyano-lH-indol-S-yl)cyclopropane-carboxamide
[00901] (Z)- l-(Benzo[d][ 1 ,3]dioxol-5-yl)-N-(2-/ert-buty]-3-((hydroxyimino)-methyI)-lH-indol-5-yl)cyclopropanecarboxamide (39 mg, 0.090 mmol) was dissolved in acetic anhydride (1 mL) and heated at reflux for 3 h. The mixture was cooled in an ice bath and the precipitate was collected and washed with water. The solid was further dried under high vacuum to yield 1 -(benzo|d][ l,3]dioxol-5-yl)-N-(2-/eri-butyl-3-cyano- 1 H-indol-5-y])cyclopropanecarboxamide. ESI-MS tnz calc.401.5, found 402.5 (M+l)+; retention time 3.70 minutes. Ή NMR (400 MHz, DMSO-d6) δ 11.72 (s, IH), 8.79 (s, 111), 7.79 (s, I II), 7.32 (in, 2H), 7.03-7.02 (ni, IH), 6.95-6.89 (m, 2H), 6.03 (s, 2H), 1.47 (s, 9H), 1.43-1.41 (ni, 2H), 1.06-1.04 (m, 2H).
[00902] Example 85: l-(Benzo[d][l,3]dioxol-5-yl)-N-(2-^rt-butyl-3-methyl-lH.indol-5-yDcyclopropanecarboxamidc
[00903] Λ solution of l-(ben7,o[d][l,3Jdioxol-5-yl)-/V-(2-iiiri-butyI-lH-indol-5-yl)cyclopropanecarboxamide (75 mg, 0.20 mmol) and iodomethane (125 μΐ,, 2.0 mmol) in NjV-dimethylformamide (1 mL) was heated at 120 °C in a sealed lube for 24 h. 'Hie reaction was filtered and purified by reverse phase I 1PLC. ESI-MS ni/z calc. 390.5, found 3 1.3 (M+l)+; retention time 2.04 minutes. Ή NMR (400 MHz, DMSO-d6) δ 10.30 (s, 1H), 8.39 (s, 1 H), 7.51 (m, 1 H), 7.1 -7. 1 1 (m, 1 H), 7.03-6.90 (m, 4H), 6.03 (s, 211), 2.25 (s, 3H), 1.40-1.38 (ni, 1111), 1.03- 1.01 (m, 2H).
[00904] Example 86: 1-(Benzo[d]tl ,3]dioxol-5-yl)-N-(2-i
[00905] Approximately 100 μΐ, of ethylene dioxide was condensed in a reaction tube at -78 °C. A solution of l-(benzo[d]| 1 ,3Jdioxol-5-yl)-N-(2-reri-butyl- lH-indol-5-yl)cyclopropanecarboxamide (200 mg, 0.50 mmoi) and indium trichloride (20 mg, 0.10 nimol) in dich!oromethane (2 mL) was added and the reaction mixture was irradiated in the microwave for 20 min at 100 °C. The volatiles were removed and the residue was purified by column chromatography (0- 100 % ethyl acctatc hcxanes) to give l -(benzo[dl[l ,3]dioxol-5-yl)-Ar-(2-icrf-butyl-3-(2-hydroxyelhyl)- l l l-indol-5-yl)cyclopropanecarboxamide (5 mg, 3%). ESI-MS tn/z calc. 420.5. found 421.3 (M+l )+; retention time 1 .67 minutes. Ί I NMR (400 MHz, CD.iCN) 6 8.78 (s, 1 H), 7.40 (in, I II), 7.33 (s, 111), 7.08 (m, 111), 6.95 - 6.87 (m, 311), 6.79 (m, 1H), 5.91 (s, 2H), 3.51 (dd, J = 5.9, 7.8 Hz, 211), 2.92 - 2.88 (m, 21-1), 2.64 (t, J = 5.8 Hz, 1H), 1.50 (m, 2H), 1. 1 (s, 9H), 1.06 (m, 2H).
[00906] Example 87: 2-(S-(l -(Benzo[d][l ,3]dioxol-5-yl)cycIopropanecarboxamido)-I H-indol-2-y!)acetic acid
[00907] To a solution of ethyl 2-(5-( l -(benzo[d][ l,3]dioxol-5-yl)cyclopropanecarboxamido)- lH-indol-2-yl)acctate (0.010 g, 0.025 mmol) in THF (0.3 mL) were added Li0H.H2O (0.002 g, 0.05 mmol) and water (0.15 mL) were added. The mixture was stirred at room temperature for 2 h. dichloromethane (3 mL) was added to the reaction mixture and the organic layer was washed with 1 N HCl (2 x 1.5 mL) and water (2 x 1.5 mL). The organic layer was dried over Na2SC>4 and filtered. The filtrate was evaporated under reduced pressure to give 2-(5-( 1 -(benzo[d][ 1 ,3jdioxol-5-yl) yelopropanecarboxamido)- 1 H-indol-2-y])-acetic acid. Ή NMR (400 MHz, DMSO-d6) δ 12.53 (s, 1 H), 10.90 (s, 1 H), 8.42 (s, 1H), 7.57 (s, 1 H), 7.17 (d, J = 8.6 Hz, 1 H), 7.05-6.90 (m, 4H), 6.17 (s, 1H), 6.02 (s, 2H), 3.69 (s, 2H), 1.41- 1.39 (m, 211), 1.04- 1.02 (m, 211).
[00908] Example 88: 5-(l-(Benzo[rf][l,3]dioxol-5-yl)cyclopropanecarboxamido)-2-
[00909] Methyl 5-( 1 -(benzol d\\ 1 ,3]dioxol-5-yl)cyclopropanecarboxamido)-2-½r/-butyl-l//-indole-7-carboxylate (30 mg, 0.069 mmol) was dissolved in a mixture of 1 ,4-dioxane (1.5 mL) and water (2 mL) containing a magnetic star bar and lithium hydroxide (30 mg, 0.71 mmol). The resulting solution was stirred at 70 °C for 45 minutes. The crude product was then acidified with 2.6 M hydrochloric acid and extracted three times with an equivalent volume of dichloromethane. The dichloromeihane extracts were combined, dried over sodium sulfate, filtered, and evaporated to dryness. 'Hie residue was dissolved in a minimum of N.N-dimethyllbrmamide and then purified by preparative HPLC using a gradient of 0-99% acetonitrile in water containing 0.05% trifiuoroacetic acid to yield 5-( l-(benzo[f/|L 1,3 Jdioxol-5-yl)cyclopropanecarboxamido)-2-/i;ri-butyl- 1 / -indole-7-carboxylic acid. ESI-MS in/z calc. 434.2, found 435.5. Retention time of 1.85 minutes. Ή NMR (400 MHz, DMSO-i/6) δ 13.05 (s, 1H), 9.96 (d, J = 1 .6 Hz, 1 H), 7.89 (d, J = 1.9 Hz, 1 H), 7.74 (d, J = 2.0 Hz, 1H), 7.02 (d, J = 1.6 Hz, 1 H), 6.96-6.88 (m, 211), 6.22 (d, J = 2.3 Hz, I H), 6.02 (s, 2H), 1.43 - 1.40 (m, 2H), 1.37 (s, 9H), 1.06- 1.02 (m, 2H).
[00910] Example 89: l-(Benzo[i/][l,3]dioxol-5-yI)-yV-(2-/ert-butyl-I-(l,3-dihydroxypropan-2-yl)-I//-indol-5-yl)cyclopropanecarboxamide [00911 ] l-(Benzo[ti][l,3]dioxol-5-yl)-N-(2-^rt-butyl-l-(l ,3-dihydroxypropan-2-yl)indolm-5-y])cyclopropanecarboxamide
[00912] l-(BenzordJ| l ,3]dioxol-5-yl)-^-(2-/ert-bulylindolin-5-yl)cyclopropanccarboxamide {50 mg, 0. 13 mmol) was dissolved in dichloroelhane (0.20 mL) and 2,2-dimethyl- l ,3-dioxan-5-one (0.20 mL). Trifluoroacetic acid was added (0.039 mL) and the resulting solution was allowed to stir for 20 minutes. Sodium triacetoxyborohydride was added (55 mg, 0.26 mmol) and the reaction mixture was stirred lor 30 minutes. The crude reaction mixture was then evaporated to dryness, dissolved in NN-dimethylformamide and purified by preparative HPLC using a gradient of 0-99% acetonitrile in water containing 0.05% trifluoroacetic acid.
[00913] l-(Benzo[rf][lT3]dioxol-S-y!)- V-(2-iert-butyl-l-(l,3-dihydroxypropa-i-2-yl)-l/ -indol-5-yl)cyclopropanecarboxamide
[00914] l-(Benzo[rf][ l ,3]dioxol-5-yi)-N-(2-ie^buLy]- l-(l,3-dihydroxypropan-2-yl)indolin-5-yl)cyclopropanccarboxamidc (40.3 mg, 0.071 1 mmol as the trifluoracetic acid salt) was dissolved in toluene ( 1 mL). To the resulting solution was added 2,3,5,6-tetrachlorocyclohexa-2,5-diene- 1,4-dione (35 mg, 0. 14 mmol). The resulting suspension was heated at 100 °C in an oil bath for 10 minutes. The crude product was then evaporated to dryness, dissolved in a 1 mL of NN-dimethylformamide and purified by purified by preparative HPLC using a gradient of 0-99% acetonitrile in wuter containing 0.05% trifluoroacetic acid to yield l -(benzo|
[00915] Example 90: /V-(7-(Aminomethyl)-2-/ert-butyl-l/ -indol-5-yI)-l -(b€nz [J][ l,3]-dioxol-5-yl)cyclopropanecarboxamide
[00916] ^-(7-(Aminomethyl)-2-tert-butyl- lH-indol-5-yl)-l -(benzotrf][l,3]dioxoi-5-yl)cyclopropanecarboxamide
[00917] l -(Benzo[i ][ l ,3 ]dioxol-5-yl)-N-(2-/iirr-butyl-7-cyano- 1 //-indol-5-yl)cyclopropanccarboxamidc (375 mg, 0.934 mmol) was dissolved in 35 mL of ethyl acetate. Ί e solution was recirculated through a continuous How hydrogenation reactor containing 10% palladium on carbon at 100 °C under 100 bar of hydrogen for 8 h. The crude product was then evaporated to dryness and purified on 12 g of silica gel utilizing a gradient of 0-100% ethyl acetate (containing 0.5% triethyl amine) in hexanes lo yield N-(7-(aminomethyl)-2-/e^butyl H-indol-5-y])- l -(benzo[rf|l l,3]-dioxol-5-yl)-cyclopropanecarboxamide (121 mg, 32%). KSI-MS m z calc. 405.2, found 406.5 (M+ l)+. Retention time of 1.48 minutes.
[00918] Example 91 : 5-(l -(Benzo[< }[l,3]dioxol-5-yl)cycIopropanecarboxamido)-2-½rr-butyl-l//-indole-7-carboxamide
[00919] 5-(l -( Benzo[rf][l ,3]dioxol-5-yl)cyc]opropanecarboxamido)-2-tor/-butyl- 1H-indole-7-carboxamide
[00920] 1 -(Benzoyl 1 ,3]dioxol-5-yl)-jV-(2-/m-butyl-7-cyano- 1 tf-indol-5-yl)-cyclopropanecarbox amide (45 mg, 0. 1 1 mmol) was suspended in a mixture of methanol ( 1.8 mL), 30% aqueous hydrogen peroxide (0.14 mL, 4.4 mmol) and 10% aqueous sodium hydroxide (0.150 mL). The resulting suspension was stirred for 72 h at room temperature. JTie hydrogen peroxide was then quenched with sodium sulfite. The reaction mixture was diluted with 0.5 mL of NN-dimethylformamide, filtered, and purified by preparative HPLC using a gradient of 0-99% acetonitrile in water containing 0.05 % trifluoroacetic acid to yield 5-( l -(benzo[i ][ l,31dioxol-5-yl)cyclopropane-carboxamido)-2-ieri-butyl- l //-indole-7-carboxaniide. ES1-MS m/z calc. 41 .2, found 420.3 (M+ l )+. Retention lime of 1.74 minutes.
[00921] Example 92: l-(Benzo[i ][lt3]dioxol-S-yl)-N-(2-½rt-butyl-7- (methytsulfonamido-methyl)-l//-indol-5-y])cyclopropanecarboxamide
[00922] l -(Benzo[i ][l ,3]dioxol-S-yl)-N-(2-/ert-butyl-7-(methylsulfoiiamidomethyl)-lH-indol-5-yl)cyclopropanecarboxamide
[00923] V-(7-(Aminomelhyl)-2-iirrt-butyl- 1 W-indol-5-yl)- 1 -(benzo[< ]| 1 ,3 |dioxol-5-yl)cyclopropanecarboxamide (20 mg, 0.049 mmol) was dissolved in DMF (0.5 mL) containing trielhylamine (20.6 μΐ^, 0. 147 mtnol) and a magnetic stir bar. Methanesulfonyl chloride (4.2 μί, 0.054 mmol) was then added to the reaction mixture. The reaction mixture was allowed lo stir for 12 h at room temperature. The crude product was purified by preparative I I PLC using a gradient of 0-99% acetonitriic in water containing 0.05% txifluoroacetic acid to yield l-(benzo|i/J [ l ,3]dioxol-5-yl)-N-(2-½ri-butyl-7-(methylsulfonamidomethyI)- l f/-indo]-5-yl)cyclopropanecarboxamide. ESI-MS m/z calc. 483.2, found 484.3 (M+ l )+. Retention ti me of 1 .84 minutes.
[00924] Example 93: Ai-(7-(Acelamidomet yl)-2-tert-butyl-1ff-indol-5-yl)-l -(benzo[rf][l ,3]-dioxol-5-yl)cyclopropanecarrJoxamide
[00925] Af-(7-(Aminomethyl)-2-ier/-butyl- l /-indol-5-yl)-l -(benzo| rf]l l ,3]dioxol-5-yl)cyclopropanecarboxaniide (20 mg, 0.049 mmol) was dissolved in DMF (0.5 mL) containing trielhylamine (20.6 μΐ -, 0. 147 mmol) and a magnetic stir bar. Acetyl chloride (4.2 μ]_., 0.054 tnniol) was then added to the reaction mixture. The reaction mixture was allowed to stir for 16 h at room temperature. The crude product was purified by preparative MPLC using a gradient of 0-99% acetonitriic in water containing 0.05 % trifluoroacetic acid lo yield W-(7-(acelanndomethyl)-2-ferr-butyl-l//-indol-5-yl)- l -(benzo|iil| l,3 )dioxoU5-yl)cyclopropanecarboxamide. ESI-MS m z c lc. 447.2, found 448.3 (M+ l )+. Retention time of 1.76 minutes.
[00926] Example 94: N-(l-Acetyl-2.terr-buty|.l H-indol-S-yl)-l -(benzo[d][l,3]dioxol-5-yl)-cyclopropanecarboxamide
[00927] To a solution of I -(benzo[d]l 1 ,3]dioxol-5-yl)-N-(2-/er/-butyl- 1 H-indol-5-yl)cyclopropanecarboxamide ( 120 mg, 0.31 mmol) in anhydrous DMF-THF (3.3 inL, 1:9) was added NaH (60% in mineral oil, 49 mg, 1.2 mmol) at room temperature. After 30 min under N2, the suspension was cooled down to ~l 5 "C and a solution of acetyl chloride (1.1 eq.) in D F (0.5 niL) was added dropwise. The reaction mixture was stirred for 30 min at -15 °C then for 6 h at room temperature. Water (0.5 niL) was added at 0 °C, solvent was removed, and the residue was diluted with MeOH, filtrated and purified by preparative HPLC to give N-(l -acetyl-2-/cri-butyl- l H-indol-5-yl)-l -(benzotd] [l ,3]dioxol-5-yl)cyclo-propanecarboxaraide. Ί I NMR (400 MHz, DMSO) δ 8.9 (s, 1 H), 7.74 (d, J =2. 1 Hz, 1 H), 7.54 (d, J =9.0 Hz, 1 H), 7.28 (dd, J J =2. 1 Hz, J2 =9.0 Hz, 1 H), 7.01 (d, ./ =1.5 Hz, 1 H), 6.93 (dd, J J = 1.7 Hz, 32 =8.0 Hz, 1 11), 6.89 (d, 7 =8.0 Hz, 1 H), 6.54 (bs, H I), 6.02 (s, 2H), 2.80 (s, 3H), 1 .42- 1 .40 (m, 1 lH), 1.06- 1.05 (m, 2H). MS (IZSl) m e (M+H+) 419.3.
[00928] Example 95: N-(l-(2-Acelamidoethyl)-2-½rt-butyl-6-fluoro-i H-indol-5-yl)-l-(2,2-dif1iiorobenzo| d]ll,3]dioxol-5-yl)cyclopropanecarboxamide
[00929] N-(l-(2-Aminoethyi).2-iirt-buty]-6-fluoro-lH-indol-5-yl)-l-(2,2-difluorobertzo-[d]f1,3]dioxoI-5-yl)cydopropanecarboxamide
[00930] To a solution of/ert-butyl 2-(2-/ert-butyl-5-(l-(2,2-dinuorobcnzo[d][l,3]dioxol-5-yl)t'yctopropanecarboxamido)-6-fluoro-lH-indol-l-yl)ethylcarbamatc (620 ing, 1.08 mmol) in CH2G2 (8 mL) was added TFA (2 mL). The reaction was stirred at room temperature for 1.5 h before being neutralized with solid NaIIC03. The solution was partitioned between J-l20 and CI I2CI2. The organic layer was dried over MgSO-i, filtered and concentrated to yield the product as a cream colored solid (365 mg, 71%). Ή NMR (400 MHz, DMSO-d6) δ 8.38 (s, 1H), 7.87 (br s, 3H, NH3+), 7.52 (s, 1 H), 7.45-7.38 (m, 3H), 7.32 (dd, J - 8.3, 1.5 Hz, 1H),6.21 (s, 1H),4.46 (m, 2H), 3.02 (m, 2H), 1.46 (m, 2H), 1.41 (s, 9H), 1.14 (m, 2H). HPLC ret. time 1.66 min, 10-99 % CI l CN, 3 min run; ES1-MS 474.4 m/z (M+H+).
[00931] /V-(l-(2-Acetamidoethyi)-2-tert-butyl-6-nuoro-lH-indol-5-yl)-l-{2f2-difluorobenzo [d][l ,3]dioxol-5- l)cyclopropanecarboxamide
[00932] To a solution of N-(l-(2-aminoethyl)-2-/er/-butyl-6-fluoro-lH-indol-5-yl)-l-(2,2-difluorobenzo|d|| l,3jdioxol-5-yl)cyclopropane-carboxamide (47 mg, 0.10 mmol) and Et3N (28 ΐ., 0.20 mmol) in DMF (1 mL) was added acetyl chloride (7.1 μΐ^ 0.10 mmol). The mixlure was stirred at room temperalure for 1 h before being i'illered and purified by reverse phase HPLC (10 - 99 % CH3CN/ H20) to yield ^-(l-(2-acetamidoethyl)-2-iert-butyl-6-nuoro-lH-indol-5-yl) -(2,2-dinuorobenzo|d]| l,3]dioxol-5-yl)cyclopropanecarboxamide. Ή NMR (400 MHz, DMSO-d6) δ 8.35 (s, 1H), 8.15 (t, J = 5.9 Hz, 1H), 7.53 (s, 1H), 7.43-7.31 (m, 4H), 6.17 (s, 1H), 4.22 (m, 2H), 3.30 (m, 211), 1.85 (s, 3H), 1.47 (m, 2H), 1.41 (s, 9H), 1.13 (ra, 2H). HPLC ret. time 2.06 min, 10-99 % CH3CN, 3 min run; ESI-MS 516.4 mz (M+If).
[00933] Example 96: l-(Benzo[d][l,3]dioxol-S-yl)-iV-(2-tert-butyl-l-(2-hydroxy-3-methoxy-propyI)-lH-indol-5-yl)cycIopropenecarboxamide
[00934] 1 -(Benzold][ 1 ,3]dioxu!-5-yl)-AK2-/«?rf-butyl- l H-indol-5-yl)cyclopropanecarboxamide (320 mg, 0.84 mnio!) was dissolved in a mixture composed of anhydrous DMF (0.5 mL) and anhydrous THF (5 mL) under N2. Nai l (60% in mineral oil, 120 nig, 3.0 mmol) was added at room temperature. After 30 min of stirring, the reaction mixture was cooled to - 15 °C before a solution of epichlorohydrin (79 ί, 1 .0 mmol) in anhydrous DMF ( 1 mL) was added dropwise. The reaction mixture was stirred for 15 min at -15 CC, then for 8 h at room temperature. MeOH (1 mL) was added and the mixture was heated for 10 min at 105 °C in the microwave oven. The mixture was cooled, filtered and purified by preparative HPLC to give 1 -(benzol dj[ 1 , ]dioxol-5-yl)-jV-(2-?i?rt-bulyl- l-(2-hydroxy-3-methoxy-propyl)- 1 1 l-indol-5-yl)cyclopropanecarboxamide. Ή NM (400 MHz, D SO-d6) 8 8.44 (s, 1 H), 7.59 (d, J = ) .9 Hz, H I), 7.31 (d, J = 8.9 Hz, 1 H), 7.03 (dd, J = 8.7, 1 .9 Hz, 2H), 6.95 (dd, 7 = 8.0, 1.7 Hz, 1 I I), 6.90 (d, J = 8.0 Hz, 1 H), 6.16 (s, 1H), 6.03 (s, 211), 4.33 (dd, J = 15.0, 4.0 Hz, 111), 4.1 (dd, J = 15.0, 8.1 Hz, 1 I I), 4.02 (ddd, J = 8.7, 4.8 Hz, H I), 3.41 -3.32 (m, 2H), 3.30 (s, 311), 1.41 (s, 9H), 1.41- 1.38 (m, 2H), 1 .03 (dd, 7 = 6.7, 4.0 Hz, 2H). MS (ESI) m/e (M+H+) 465.0.
[00935] Example 97: l-(Benzo[d)[ ]dioxol-5-yl)-Ar-(2-/ert-butyl-1-(2-hydroxy-3-(met yl-ainino)propyl)-lH-indol-5-yl)cyclopropanecarboxamide
[00936] 1 -(Benzo[d] [1 ,3]dioxol-5-y l)-N-(2-ier/-butyl- 1 H-indol-5-yl)cyclopropanecarboxamide (320 mg, 0.84 mmol) was dissolved in a mixture composed of anhydrous DMF (0.5 mL) and anhydrous THF (5 mL) under N2. NaH (60% in mineral oil, 120 mg, 3.0 mmol) was added at room temperature. After 30 min of stirring, the reaction mixture was cooled to -15 °C before a solution of epichlorohydrin (79 \iL, 1.0 mmol) in anhydrous DMF ( 1 mL) was added dropwise. The reaction mixture was stirred for 15 min at - 15 °C, then for 8 h at room temperature. MeNH2 (2.0 M in MeOH, 1.0 mL) was added and the mixture was heated for 10 min at 105 °C in the microwave oven. The mixture was cooled, filtered and purified by preparative HPLC to give l -(benzo|d || l ,31dioxol-5-yl)-N-(2-ierr-butyl- 1 -(2-hydroxy-3-(melhylamino)propyl)- 1 H-indo]-5-yl)cyclopropanecarboxamide. Ή NM R (400 MHz, DMSO-d6) 8 8.50 (s, I II), 7.60-7.59 (m, 111), 7.35 (dd, J = 14.3, 8.9 Hz, 1 H), 7.10 (d, J = 8.8 Hz, 1H), 1H), 6.94 (dd, J = 8.0, 1.6 Hz, 1H), 6.91 (d, J = 7.9 Hz, 1H), 6.20 (d, J = 2.3 Hz, 1H), 6.03 (s, 2H), 2.82 (d, J = 4.7 Hz, 1H), 2.72 (d, J = 4.7 Hz, HI), 2.55 (dd, J = 5.2, 5.2 Hz, 1H), 2.50 (s, 3H), 1.43 (s, 911), 1.39 (dd, J = 6.4, 3.7 Hz, 211), 1.04 (dd, J = 6.5, 3.9 Hz, 211). MS (ESI) m/e (M+H+) 464.0.
[00937] Example 98: (S)-iV-(l-(3-Amino-2-hydroxypropyl)-2-¾rt-butyl-lH-indol-5-yl)-l-(2,2-difluorobenzold][l,3]dioxol-5-yl)cyclopropanecarboxamide
[00938] («)-3-(2-/ert-Butyl-5-(l-(2,2-dinuorobenzo[d][l,3]dioxol-5-yl)cyclopropanecarbox-amido)-l H-indol-1 -yl)-2-hydroxypropyl-4-methylbenzenesulfonate
[00939] To a stirred solution of (/f )-N-(2-/ert-butyl- 1 -(2,3-dihydroxypropy 1)- 1 H-indol-5-yl)-l-(2,2-difluoro-benzo|d]tl,3|dioxol-5-yl)cyclopropanecarboxamide (3.0 g, 6.1 mmol) in dichloro methane (20 niL) was added tri ethyl amine (2 niL) and para-toluenesulfonylchloride (1.3 g, 7.0 mmol). After 18 hours, the reaction mixture was partitioned between 10 niL of water and 10 mL of ethyl acetate. The organic layer was dried over magnesium sulfate, filtered and evaporated. The residue was purified using column chromatography on silica gel (0-60% ethyl acetate/hexane) providing (/i)-3-(2-fer/-butyl-5-(l-(2,2-difluorobenzo[d][l,3]-dioxol-5-yl)cyclopropanccarboxamido)- 1 H-indol- l-yl)-2-hydroxypropyl-4-methyl-benzenesulfonate (3.21 g, 86%). LC/MS (M + 1) = 641.2. 1H NMR (400 MHz, CDCI3) 8 7.77 (d, 2H, 7=16 Hz), 7.55 (d, 1H, J = 2 Hz), 7.35 (d, 211, J = 16 Hz), 7.31 (m, 3H), 6.96 (s, 111), 6.94 (dd, 1H, J = 2, 8 Hz), 6.22 (s, 1H), 4.33 (m, III), 4.31 (dd, 1H,7 = 6, 15 Hz), 4.28 (dd, 111,./= 11, 15 Hz).4.18 (in, 1H),3.40 (dd, 1H, ./ = 3, 6 Hz), 3.36 (dd, 1H, ,/ = 3, 6 Hz), 2.46 (s,3H), 2.40 (brs, IH), 1.74 (m,2H), 1.40 (s,9H), 1.11 (m, 2 H).
[00940] {K)-N-(l-{3-Azido-2-hydroxypropyl)-2-½r(-butyl-lH-indol-5-yl)-l-(2,2-difluorobenzo [d][l ,3]diox l-5-yl)oyclopropatiecarboxamide
[00941] To a stirred solution (-¾)-3-(2-/erf-butyl-5-(l-(2,2-difluorobenzo[dJ[l,3Jdioxol-5-yl)cyclopropanecarboxamido)- 111-indol-l -yl)-2-hydroxypropyl-4-inethylbcnzenesulfonate (3.2 g, 5.0 mmol) in DMF (6 mL) was added sodium azide (2.0 g, 30 niniol). The reaction was heated at 80 °C for 2 h. The mixture was partitioned between 20 mL ethyl acetate and 20 mL water. The layers were separated and the organic layer was evaporated. rHie residue was puri ied using column chromatography (0-85% ethyl acetate/hexane) to give (/i)-N-(l-(3-azido-2-hydroxypropyl)-2-/m-butyl H-indol-5-yl)-l-(2,2-dinuorobenzo[d]|'l,3]dioxol-5-yl)-cyclopropanecarboxamide (2.48 g). LC/MS ( + I) = 512.5. Ή NMR (400 MHz, CDCI3) δ 7.55 (d, 1H, 7 = 2 Hz), 7.31 (m, 3H), 6.96 (s.1H), 6.94 (dd, 1H, ./ = 2, 8 Hz), 6.22 (s, 1H), 4.33 (m, 111), 4.31 (dd, 1H,7 = 6, 15 Hz), 4.28 (dd, lH,/ = 11, 15 Hz), 4.18 (m, IH), 3.40 (dd, lH,1/ = 3,6Hz),3.36(dd, III, ,/ = 3, 6 Hz), 2.40 (brs, 1H), 1.74 (m, 2H), 1.40 (s, 9H), 1.11 (m,2H).
[00942] (S)-N-(l-(3-Amino-2-hydroxypropyl)-2-tert-butyl-lH-indol-5-yl)-l-(2,2-difluoro-benzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamide
[00943] To a stirred solution (?)-V-(l-(3-azido-2-hydroxypropyl)-2-/fc-r/-butyl-lH-indol-5-yl)-l-(2,2-difluorobenzo [d|{l,3Jdioxol-5-yI)cyclopropanecarboxamide (2.4 g, 4.0 mmol) in MeOH (25 mL ) was added 5 % Pd/C (2.4 g) under a Hydrogen gas filled balloon. After 18 h, the reaction mixture was filtered through celite and rinsed with 300 mL ethyl acetate. The organic layer was washed with 1 N IICl and evaporated to give (5)-/V-(l-(3-amino-2-hydroxypropyl)-2-f
[00944] Example 99: (S)-Methyl 3-(2-rert-butyl-5-(l-(2,2-dinuorobenzo[d]tl,3]dioxol-5-yl)cyclopropanecarboxamido)-lH-indol-l-yl)-2-hydroxypropylcarbamate
[00945] To a stirred solution (?)-N-(l-(3-amino-2-hydroxypropyl)-2-/er/-butyl-lH-indol-5-yl)-l-(2,2-difluorobenzo|d|[ l,3]dioxol-5-yl)cyclopropanecarboxamide (0.10 g, 0.20 mmol) in methanol (1 mL) was added 2 drops of triethylamine and methylchJoroformyl chloride (0.020 mL, 0.25 mmol). After 30 min, the reaction mixture was filtered and purified using reverse phase HPLC providing (A1)- methyl 3-(2-/eri-bulyl-5-(l-{2,2-dilluorobeiizo[d]| l,3]dioxol-5-yl)cyclo-propanecarboxamido)- 1 H-indol- 1 -yl)-2-hydroxypropylcarbaniate. The retention time on a three minute run is 1.40 min. LC/MS (M + 1) = 544.3. Ή NM (400 MHz, CD 13) δ 7.52 (d, lH, J = 2Hz), 7.30 (dd, 1H, /= 2, 8 Hz), 7.28(m, 1H), 7.22 (d, 1H, 7 = 8 Hz), 7.14 (d, lHs = 8 Hz), 7.04 (brs, 111), 6.97 (dd, 1Η,/ = 2, 8 Hz), 6.24 (s, 1H), 5.19 (1H, br s), 4.31 (dd, IH, J = 6, 15 Hz), 4.28 (dd, 111, J= 11, 15 Hz), 4.18 (m, 1H), 3.70 (s, 3H), 3.40 (dd, 1H,./ = 3, 1-Iz),3.36(dd, 111, ./= 3, 6 Hz), 3.26 (m, 1H).1.74 (in, 2H), 1.40 (s, 9 H), 1.11 (m,2H).
[00946] Example 100: 4-(5-(l-(Benzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamido)-2-tert-butyl-lH-indol-1 - Pbutanoic acid
[00947] l-(Benzo[d][l ,3]dioxol-5-yI)-Ar-(2-½rt-butylindolin-5-y I)cy clo propanecar bo xamide
[00948] To a solution of l-(benzo[d][ l ,3]dioxol-5-yl)-W-(2-terf-bulyl- l H-indo]-5-yl)cyclo-propanecarboxamide (851 mg, 2.26 mmol) in acetic acid (60 ml.,) was added NaBH3CN (309 mg, 4. 1 mmol) at 0 °C. 'ITie reaction mixture was stirred for 5 min at room temperature after which no starting material could be delected by LC S. The solvent was evaporated under reduced pressure and the residue was purified by column chromatography on silica gel (5-40% ethyl acetate/hexanes) to give l-(benzo[d || l ,3Jdioxol-5-yl)-A'-(2-ieri-butylindolin-5-yl)cyclopropanecarboxamide (760 mg, 89%).
[00949] 4-(5-(] -(Benzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamido)-2-^rt-butylindolin-l-vDbutanoic acid
[00950] To a solution of l -(benzo|d|| l ,3]dioxol-5-yl)-Ar-(2-/i;ri-butylindolin-5-yDcyclopropanecarboxamide (350 mg, 0.93 mmol, 1 eq) in anhydrous methanol (6.5 niL) and AcOl l (65 μΙ„) was added 4-oxobutanoic acid ( 15% in water, 710 mg, 1 .0 mmol) at room temperature. After 20 min of stirring, NaBH3CN (130 mg, 2.0 mmol) was added in one portion and the reaction mixture was stirred i'or another 4 h at room temperature. Ί he reaction mixture was quenched by addition of AcOH (0.5 mL) at 0 °C and the solvent was removed under reduced pressure. 'ITie residue was purified by column chromatography on silica gel (5-75% ethyl acetate hexanes) to give 4-(5-(l-(benzo[dj| l,3]dioxol-5-yl)cyclopropanecarboxamido)-2-/eri-butylindolin- l -yl)butanoic acid ( 130 mg, 30%).
[00951] 4-(5-(l-(Benzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamido)-2-/i?rt-butyl-lH-indol-l-yl)butanoic acid
[00952] 4-(5-( ! -(Benzo|d|| 1 ,3]dioxok5-yl)cyclopropaneearboxaniido)-2-rer/-butylindolin-l -yl)bulanoic acid (130 mg, 0.28 nimol) was taken up in a mixture of acetonitrile-I O-TFA. ITie solvent was removed under reduced pressure and the residue obtained was dissolved in CDC¾. After a brief exposition to daylight (5- 10 min), the solution turned purple. The mixture was stirred open to the atmosphere at room temperature until complete disappearance of the starting material (8 h). Solvent was removed under reduced pressure and the residue was purified by reverse pharse HPLC to give 4-(5-(l -(benzo[dl| l ,3Jdioxol-5-yl)cyclopropanecarboxamido)-2-tert-butyl-lH-indol-l-yl)butanoic acid. l NMR (400 MHz, CDC13) 6 7.52 (d, J = 1.9 Hz, 1H), 7.18 (d, J = 2.1 Hz, l H), 7.16 (s, 1 H), 7.03 (dd, J = 9.4, 1.9 Hz, 111), 7.00-6.98 (m, 211), 6.85 (d, .1 = 7.9 Hz, 111), 6.1 (s, 1H), 6.02 (s, 2H), 4.29-4.24 (m, 2H), 2.48 (dd, J - 6.9, 6.9 Hz, 2H), 2.12-2.04 (m, 2H), 1.69 (dd, J = 6.8, 3.7 Hz, 2H), 1.43 (s, 9H), 1.09 (dd, J = 6.8, 3.7 Hz, 2H). MS (ESI) m/c (M+H+) 463.0.
[00953] Example 101 : l-(Benzo[d][l,3]dioxol-5-y])-A'-(2-tert-butyl-l-(4-(2-hydroxy ethyl-amino)- 4-oxobuty 1)- 1 H - indol-5 -y l)cy clopropanecar bo xamide
[00954] To a solution of 4-(5-(l-(benzo[d][ l ,3]dioxol-5-yl)cyclopropanecarboxamido)-2-iert-bulyl-l H-indol- l-yl)buianoic acid (10 mg) in anhydrous DMF (0.25 niL) were successively added I¾N (9.5 niL, 0.069 nimol) and HBTU (8.2 mg, 0.022 mmol). After stirring for 10 min at 60 °C, ethanolamine (1.3 ΐ^, 0.022 mmol) was added, and the mixture was stirred for another 4 h at 60 °C. l-(Benzo[d][ l ,3]dioxol-5-yl)-N-(2-½r/-butyI-l -(4-(2-hydroxyethyl-amino)-4-oxobutyl)- lH-indol-5-yl)cyclopropanecarboxamide (5.8 mg, 64%) was obtained after purification by preparative HPLC. MS (I¾I) m e (M+H+) 506.0.
[00955] Example 102: l-(Benzo[d][l>3]dioxol-5-yl)-A'-(2-/e'rt-butyl-l-(2-(dimethylatnino)-2-oxoethyl)-lH-indol-5-yl)cyclopropanecarboxamide 1. NaH, D F-THF
[00956] To a solution of l -(benzo[d]| l ,3]dioxol-5-yl)- V-(2-ieri-butylindoltn-5-yl)cyclopropanccarboxamidc (62 mg, 0.16 mmol) in anhydrous DMF (0.1 1 mL) and THF ( 1 mL) was added NaH (60% in mineral oil, 21 mg, 0.51 mmol) at room temperature under N2. After 30 min of stirring, the reaction mixture was cooled to 0 °C and 2-chloro-N,N-dime hylacetamide ( 1 1 mL, 0.14 mmol,) was added. The reaction mixture was stirred for 5 min at 0 °C and then for 10 h at room temperature. The mixture was purified by preparative HPLC and the resultant solid was dissolved in DMF (0.6 mL) in the presence of Pd-C (10 mg). The mixture was stirred open to the atmosphere overnight at room temperature. The reaction mixture was filtrated and purified by preparative HPLC providing 1 -(bcnzo[d] [ 1,3 ]dioxol-5-yl)-N-(2-rm-butyl- 1 -(2-(dimethylamino)-2-oxocthyl)- 1 H-indol-5-y])cyelopropanecarboxamide. MS (ESI) ni/e (M+H+) 462.0.
[00957] Example 103: 3-(2-«irt-Butyl-5-(l -(2,2-dinuorobenzotd]tl,3]dioxol-5-yi)cyclo-propanecarboxamido)- lH-indol-l -yl)propanoic acid
[00958] tf-(2-fcrt-Butyl-1 -(2-chloroethyl)indoliii-5-yl)-l -(2,2-dinuorobenzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamide
[00959] To u solution oi' Ar-(2-/tiri-butyl- l -(2* yano-;thyl)indolin-5-yl)-1 -(2,2-difluorobenzotdj[ l,3]dioxol-5-yl)cyclopropanecaiboxaniide (71 nig, 0.17 mniol) in anhydrous dichloromethane (1 mL) was added chloroacelaldehyde (53 μL, 0.41 inmol) al room temperature under N2. After 20 min of stirring, NaBH(OAc>3 (90 mg, 0.42 mniol) was added in two portions. The reaction mixture was stirred overnight at room temperature. The product was purified by column chromatography on silica gel (2- 15% ethyl acetate hexanes) providing N-(2-ieri-butyl- I-(2-chIoroethyl)indolin-5-yi)- l-(2,2-difluoroben/o dJ| l,3]dioxol-5-yl)cyclopropanccarboxaniide (51 mg, 63%).
[00960] N-(2-.£?rt-Butyl-l-(2-cyanoethyl)indoliii-S-yl)-l-(2,2-difluorobenzo[d][l,3]dioxol-5-yl)cyclopropanecarboxaniide [00961 ] AH2-f_Tf-butyl- 1 -(2-chloroethyl)indolin-5-yl)- 1 -(2,2-difluorobenzo[dl[ 1 ,3]dioxol-5-yl)cyclopropanecarboxamide (51 mg), NaCN (16 mg, 0.32 mmol) and I (cat) in EtOH (0.6 mL) and water (0.3 mL) were combined and heated al 1 10 "C for 30 min in the microwave. 'l"he solvent was removed under reduced pressure and the residue was purified by column chrom tography on silica gel (2- 15% ethyl acetate/hexanes) providing N-(2-/cr/-butyl- l-(2-cyanoethyl)indolin-5-yl)- l -(2,2-dii1uorobenzo[d][1 |clioxol-5-yl)cyclopropanccarboxamide (24 mg, 48%).
[00962] 3-(2-¾rt-Butyl-5-(l -(2,2-dinuorobenzo[d][l,3]dioxoI-5-yl)cyclo-propanecarbox-amido)-lH-indol-l-yl)propanoic acid
[00963] A' 2-½ri-butyl-l-(2-cyanoethyl)indolin-5-yl)- I -(2,2-dinuorobenzo[dJ[l ,3]dioxol-5-yl)cyclopropane-carboxamide (24 mg, 0.050 mmol) was taken up in 50% aq. KOH (0.5 mL) and 1 ,4-dioxane ( 1 mL). The mixture was heated at 125 °C for 2 h. The solvent was removed and the residue was purified by preparative HPLC. The residue was dissolved in CDCI3 (1 mL) then briefly exposed to daylight. The purple solution that formed was stirred until complete disappearance of the starting material (1 h). 'Hie solvent was removed under reduced pressure and the residue was purified by preparative HPLC providing 3-(2-/t'/7-butyl-5-( l -(2,2-difluorobenzold][ 1 ,3]dioxol-5-yI)cyclo-propanecarboxamido)- 1 H-indoI-I -yDpropanoic acid. MS (ESI) m/e (Μ+1 Γ) 485.0.
[00964] Example 104: l-(Benzo[d][l,3]dioxol-S-yI)- V-(2-tert-butyl-6-nuoro- l-(2-hydroxy-ethyl)-l H-indoI-5-yl)cyclopropenecarboxamide
[00965] To a solution of 1 -(benzol d"|| 1 ,3]dioxol-5-yl)-N-(2-icr/-buiy]-6-fluoroijidolin-5-yl)cyclopropanecarboxamide (340 mg, 0.86 mmol) in anhydrous McOH (5.7 mL) containing 1 % of acetic acid was added glyoxal 40% in water (0.60 mL, 5.2 mmol) at room temperature under N2. After 20 min of stirring, NaBH3CN (120 mg, 1.9 mmol) was added in one portion and the reaction mixture was stirred overnight at room temperature. The solvent was removed under reduced pressure and the residue obtained was purified by column chromatography on silica gel ( 10-40% ethyl acetate/hexanes) providing a pale yellow oil which was treated with 50/50 CH3CN-H20 containing 0.05% TFA and CDCI3. Solvent was removed under reduced pressure and the residue was purified by column chromatography on silica gel (20-35% ethyl acetate/hexanes) to give l -(benzo[d][ l ,31dioxol-5-yl)-jV-(2-½ri-buiyl-6-nuoro-l-(2-hydroxyethyl)- l I-l-indol-5-yl)cyclopropanecarboxamide. Ή NMR (400 MHz, CDCI3) δ 8.02 (d, J = 7.7 Hz, 1 H , 7.30 (d, J = 2.1 Hz, 1H), 6.93 (dd, J = 1.6, 7.9 Hz, 1 H), 6.90 (d, J = 1.6 Hz, 1H), 6.90 (d, J = 1.6 Hz, 1H), 6.78 (d, J = 7.9 Hz, 1H), 6.08 (s, 1 H), 5.92 (s, 2H), 4.21 (dd, J = 6.9, 6.9 Hz, 2H), 3.68 (m, 2H), 2.28 (s, I H), 1 .60 (dd, J = 3.7, 6.7 Hz, 2H), 1.35 - 1.32 (m, 9H), 1 .04 (dd, J = 3.7, 6.8 Hz, 2H). MS (ESI) m/e (M+H+) 439.0.
[00966] Example 105: l-(Benzo[d][l,3]dioxoI-5-yl)-/V-(2-tert-butyl-6-nuoro-l-(3-hyd roxy- prop l)-! H -indo l-5-yl )cyclopropanecar boxamide
[00967] 3-(Benzyloxy)propanaL
[00968] To a suspension of PCC (606 nig, 2.82 mmol) in anhydrous dichloromethane (8 mL) at room temperature under N2 was added a solution of 3-benzyloxy- 1 -propanol (310 mg, 1.88 mmol) in anhydrous dichloromethane. The reaction mixture was stirred overnight at room temperature, filtrated through Celile, and concentrated. The residue was purified by column chromatography on silica gel ( 1 - 10% ethyl acetale hexanes) to give 3-(benzyloxy)propanal (243 mg, 79%).
[00969] l-(Benzo[d][l,3]dioxol-S-yl)-N-(2-/ert-butyl-6 fluoro-]-(3-hydroxypropyl)-lH-indol- 5• c clopropanecarboxamide
[00970] To a solution of l-(benzo| d][ 1 ,3]dioxol-5-yl)-N-(2-/m-butyl-6-fluoroindolin-5-y cyclopropanecarboxamide ( 160 mg, 0.50 mmol) in anhydrous dichloromethane (3.4 mL) was added 3-(benzyloxy)propanal (160 mg, 0.98 mmol) at room temperature. After 10 min of stirring, Nal¾H(OAc)3 (140 mg, 0.65 mmol) was added in one portion and the reaction mixture was stirred for 4 h at room temperature. The solvent was removed under reduced pressure and the residue was taken-up in a mixture of 50/50 CH3CN-H2O containing 0.05% TFA. The mixture was concentrated to dryness and the residue was dissolved in C1X¾ (5 mL) and briefly exposed to daylight. The purple solution was stirred open to the atmosphere at room temperature for 2 h. ITie solvent was removed under reduced pressure and the residue was treated with Pd-C ( 10 mg) in MeOH (2 mL) under 1 aim of H2 for 2 h. The catalyst was filtered through Celite and the solvent was removed under reduced pressure. The residue was purified by preparative TLC 30% ethyl acclatc/hcxancs to provide 1 -(benzoldJ I l ,3]dioxol-5-yl)-A'-(2-ier/-butyl-6-nuoro- l-(3-hydroxypropyl)- lH-indol-5-yl)cyclopropanecarboxamide ( 18 mg, 8% from l -(benzo[d )[ l ,3]dioxol-5-yl)-j'V-(2-ierr-butyl-6-fluoroindolin-5-yl)cyclopropane-carboxamide). Ή NM (400 MHz, CDC13) δ 8.1 1 (d, J = 7.8 Hz, 1 H), 7.31 (d, J = 2.2 Hz, 1 H), 6.94 (dd, J = 7.9, 1.7 Hz, 1H), 6.91 (d, J = 1.6 Hz, 11 1), 6.85 (d, J = 1 1 .7 Hz, 1 H), 6.79 (d, J = 7.9 Hz, 1 H), 6.10 (s, 11-1), 5.94 (s, 211), 4.25-4.21 (m, 2H), 3.70 (dd, J - 5.7, 5.7 Hz, 211), 1.93- 1 .86 (m, 2H), 1.61 (dd, J " 6.8, 3.7 Hz, 21 1), 1 .35 (s, 9H), 1 .04 (dd, J = 6.8, 3.7 Hz, 21 1). MS (ESI) ni/c (M+H+) 453.0. [00971 ] Example 106: V-(l-(2-Acetamidoelhyl)-2-iert-butyl-lH-indol-5-yl)-l-(benzo[dHl^]-dioxol-5-yI)cyclopropanecarboxamide
[00972] N-(l -(2'azidoethyl)-2-ieri-butyl-l H-indol-S-yl)-l -(benzo[d][l ,3]dioxoI-S-yl)-cyclopropanecarboxamide
[00973] To a solution of l -(benzo|dJI l ,3]dioxol-5-yl)-A/-(2-/e«-butylindo!in-5-yl)cyclopropane-carboxamide (73 mg, 0. 1 9 mmol) in anhydrous dichloromethane ( 1 .2 ml .) was added chloroaeeialdehyde (60 , 0.24 mmol) at room temperature. After 10 mi n of stirring, NaBH(OAc)3 (52 mg, 0.24 mmol) was added in one portion and the reaction mixture was stirred for another 30 min at room temperature. The solvent was removed under reduced pressure and the residue was purified by preparative HPLC to give the indoline, which oxidized to the corresponding indole when taken-up in CDCI3. 'Hie resulting indole was treated with NaN3 (58 mg, 0.89 mmol) and Nal (cat) in anhydrous DMF (0.8 mL) for 2 h at 85 °C The reaction mixture was purified by preparative HPLC providing W-(1 -(2-azidoethyl)-2-ieri-butyl- 1 H-indol-5-yl)- l-(benzo[dj l ,3]dioxol-5-yl)cyclopropanecarboxamide ( 15 mg, 18% from l -fbenzo|'d][ l ,3]dioxoI-5-yJ)-N-(2-rerr-butylindolin-5-yl)cydopropane-carboxamide).
[00974] N.(l-(2-Acetamidoethyl)-2-/eri-butyl-lH-indol-5-yl)-l-(benzo[d]tl,3]-dioxol-5-y l)cy clopropanecar boxami de
[00975] Λ solution ol" fV-( l -(2-azidoe.hyl)-2-/ert-butyl- l H-indol-5-yl)-l -{benzo[d][ l,3]dioxol-5-yl)cyclopropanecarboxamtde (13 mg, 0.029 mmol) in MeOH-AcOH (0.2 mi., 99: 1) in the presence of Pd-C (2 mg) was stirred at room temperature under 1 atm of H2 for 2 h, filtered through Celite, and concentrated under reduced pressure. The crude product was treated with AcCl (0.05 mL) and Et3N (0.05 mL) in anhydrous THF (0.2 mL) at 0 °C for 30 min and then 1 h at room temperature. l¾e mixture was purified by preparative HPLC providing N-( l-(2-acetainidoethyl)-2-/e?ri-butyl- 1 H-indol-5-yl)- 1 -(benzo[d )| 1 ,3 ]-dioxo1-5-yl)cyc]opropanecarboxaniide. MS (ESI) m/e (Μ+ΐ ) 462.0.
[00976] Example 107: iV-(2-tert-Butyl-l-(3-cyano-2-hydroxypropyl)-lH-indot-5-yl)-l-(2,2-difluorobenzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamide
[00977] 3-(2-tert-Butyl-5-(l -(2,2-difluorobenzo[d][lr3]dioxo]-5-yl)cyclopropanecarbox-amido)-lH-indol-l -yl)-2-hydroxypropyl-4-methylbenzenesulfonate
[00978] To a solution of N-(2-½rf-bulyl-l-(2,3-dihydroxypropyl) M-indol-5-yl)-l -(2,2-difluorobenzoLd]| l ,3j-dioxol-5-yl)cyclopropanecarboxamide (172 mg, 0.35 mmol) in anhydrous dichloromethane ( 1.4 mL) at O °C in the presence of Et3N (56 M-L, 0.40 mmol) was added TsCl (71 mg, 0.37 mmol). The reaction mixture was stirred for 2 h at room temperature before being cooled to 0 DC and another portion of TsCl (71 mg, 0.37 mmol) was added. After 1 h of stirring at room temperature, the mixture was purified by column chromatography on silica gel ( 10-30% ethyl acetate/hexanes) providing 3-(2-ierr-butyl-5-(l-(2,2-dinuorobenzo[d][ l ,3]dioxol-5-yl)cyclopropanecarboxamido)-l H-indo]-l-yl)-2-hydroxypropyl-4-methylbenzene-sulfonate (146 mg, 64%).
[00979] N-(2-fert-Butyl-l-(3-cyano-2-hydroxypropyI)-lH-indol-5-yl)-l-(2,2-difluorobenzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamide
[00980] N-(2-½ri-BiJtyl- l-(3-cyano-2-hydroxypropyl)-l H-indol-5-yI)- l -(2,2-difluorobenzoldjl l,3]dioxol-5-yl)-cyclopropanecarboxamide (145 mg, 0.226 mmol) was treaLcd with powdered NaCN (34 nig, 0.69 mmol) in anhydrous DMF ( 1.5 mL) at 85 °C Tor 2 h. The reaction mixture was cooled down lo room temperature before it was diluted with dichloromethane (10 mL) and aq. sat. NaHC >3 (10 mL). The organic phase was separated and the aqueous phase was extracted with dichloromethane (2 x 10 mL). The organic phases were combined, washed with brine, dried with sodium sulfate, filtered Lhen concentrated. Ί Tie residue was purified by column chromatography on silica gel (25-55% ethyl acetate hexanes) providing N-(2-/-T/-bulyl- l-(3-cyano-2-hydroxypropyl)- 1 H-indol-5-yI)-l -(2,2-dil1uorobenzo[d|Ll ,3]dioxol-5-yl)cyclopropanecarboxamide (89 mg, 79%). ] l l NMR (400 MHz, CDC13) δ 7.43 (d, J = 1.9 Hz, 1H), 7.20-7.16 (m, 2H), 7.08 (d, J = 8.8 Hz, 1 H), 7.04 (d, J = 8.2 Hz, 1 I I), 6.94 (s, 1H), 6.88 (dd, J = 8.7, 2.0 Hz, 1H), 6.16 (s, 1 H), 4.32-4.1 (m, 311), 2.83 (s, III), 2.40 (dd, J = 5.2, 5.2 Hz, 2H), 1.62 (dd, J = 6.6, 3.6 Hz, 211), 1.35 (s, 9H), 1.04 (dd, J = 6.9, 3.9 Hz, 2H). MS (ESI) m e (M+H+) 496.0.
[00981] Example 108: A'-(2-/ert-Butyl-l-(2-hydroxy-3-(2H-tetrazol-S-yl)propyl)-lH> indol- 5 - yl)- 1 -(2,2-difluorobenzo[d] [ 1 ,3]dioxol-5- y 1) cy clopropanecar boxamide
[00982] To a solution of N-(2-ieri-butyl- I-(3-cyano-2-hydroxypropy])- lH-indol-5-yl)- l - (2,2-difluorobenzo[dJ[ l ,3]dioxol-5-yl)cyclopropanecarboxamide (27 mg, 0.054 mmol) in anhydrous DMF (1.2 mL) were successively added NH4CI (35 mg, 0.65 mmol) and NaN3 (43 mg, 0.65 mmol) at room temperature. The reaction mixture was stirred for 4 h at 1 10 °C in the microwave, at which stage 50% of the starting material was converted lo the desired product. The reaction mixture was purified by preparative HPLC to provide /V-(2-feri-butyl- 1 -(2-hydroxy-3-(2H-tetrazol -5-yl)propyl)- 1 H-indol-5-yl)- 1 -(2,2-difluorobenzo-|d]Ll ,3]dioxol-5-yl)cyclopropanecarboxamide. MS (ESI) m/e (M+H+) 539.0.
[00983] Example 109: 4-(2-½rt-Butyl-5-(l-(2,2-dinuorobenzotd][l ,3]dioxol-5-yt)cyclo-propanecarboxamido)-lH-indol-l-yl)-3-hydroxybutanoic acid
[00984] Λ solution oi
[00985] Example 110: N-(l-(2-(2H-Tetrazol-5-yl)ethy])-2-tert-butyl-lH-indol-5.yl)-l-(benzo[d][l,3]dioxo]-5-yl)cyclopropanecarboxamide
[00986] I-(Benzo[d][1>3]dioxol-5-yl)-N-(2-i
[00987] To a solution of l -(bcnzo|d][l ,3]dioxol-5-yl)-Ar-(2-½ri-butyl- l-(2-chloroeLhyl)indolin-5-yl)cyclopropanecarboxamide (66 mg, 0, 15 mmol) in elhanol (0.8 mL) and water (0.4 mL) were added NaCN (22 mg, 0.45 mmol) and KI (cat) at room temperature. The reaction mixture was stirred for 30 niin at 1 10 °C in the microwave before being purified by column chroinalography on silica gut (5- 15% ethyl acetate hexanes) to provide 1 -(benzo[d]tl ,3Jdioxol-5-yl)-N-(2-½r/-butyt- ! -(2-cyano-ethyl)indolin-5-yl)cyclopropanecarboxamide (50 mg, 77%).
[00988] JV-(l-(2-(2H-Tetrazol-5-yl)ethyl)-2-½rt-butyl-l H-indol-5-yl)-l-{benzo[d][i ^3]dioxol-5-yl)cyc-Opropanecarboxamide
[00989] To a solution of I -(benzo[d|| 1 ,3]dioxol-5-yl)-N-(2-ii?r/-bulyl- 1 -(2-cyano-ethyl)indolin-5-yl)cyclopropanecarboxamide (50 mg, 0. 12 mmol) in anhydrous D F (2.6 mL) was added NH4C1 (230 mg, 4.3 mmol) and NaN3 (280 mg, 4.3 mmol). JTic reaction mixture was stirred for 30 min at 1 10 °C in the microwave, filtrated, and purified by preparative HPLC. The solid residue was dissolved in CDO3 (3 mL) and briefly (2 to 4 min) exposed to daylight, which initiated a color change (purple). After 2 h of stirring open to the atmosphere at room temperature, the solvent was removed and the residue was purified by preparative HPLC to give N-( l-(2-(2H-letrazol-5-yl)ethyl)-2-½rt-butyl- l H-indol-5-yl)- 1 -(benzo[d]| l ,3 |dioxol-5-yl)cyclopropanecarboxamide. MS (ESI) m/e (Μ+1-Γ) 473.0.
[00990] Example 111: 1 -(Benzo[d][l,3]dioxol-S-yl)-N-(2-½rt-butyl-6-fluoro-l-((letrahydro-2H-pyran-3-yl)methyl)-lH-indol-5-yl)cyclopropanecarboxamide 2. CDCI3 [00991 ] To a solution of l -(benzo|d] [ l ,3]dioxol-5-yl)-N-(2-ieri-bulyl-6-lluoroindolin-5-yl)cyclopropane-carboxamide ( 150 mg, 0.38 mmol) in anhydrous dichloroniethane (2.3 mL) at room temperature under N2 was added telrahydropyran-3-carbaldehyde (54 mg, 0.47 mniol). After 20 min of stirring, NaBH(OAc)3 ( 1 10 mg, 0.51 mmol) was added in one portion at room temperature. The reaction mixture was stirred for 6 h at room temperature before being purified by column chromatography on silica gel (5-20% ethyl acetate hexanes) to provide l -(benzo[d] [ l ,3]dioxol-5-yl)-N-(2-ieri-butyl-6-fluoro- l -((tetrahydro-2H-pyran-3-yl)melhyl)indolin-5-yl)cyclopropanecarboxamide (95 mg, 50%). CDCI3 was added to the indoline und the solution was ullowed to stir overnight at ambient temperature. The solution was concentrated to give l -(benzo[d |[ l ,31dioxo]-5-yl)-N-(2-/t'ri-butyl-6-nuoro- l -((tetrahydro-2H-pyran-3-yl)methyl)- I I I-indol-5-yl)cyclopropanecarboxamide. MS (ESI) m/e ( +H+) 493.0.
[00992] Example 112: l -(BenzoM[l,3]dioxol-5-yl)-N-(2-(2-hydroxypropan-2-yl)-lH-indol-5 -y l)cy clopropanecar boxa mide
[00993] Methyl 5-( l -(bun/.o[d\[ 1 ,3]dioxol-5-yl)cyclopropane-earboxamido)-l Hindole~2-carboxyl te ( 100 mg, 0.255 mmol) was dissolved in anhydrous tetrahydrofuran (2 mL) under an argon atmosphere. The solution was cooled to 0 °C in an ice water bath before methylliihium (0.85 mL, 1 .6 M in diethyl ether) was added by syringe. The mixture was allowed to warm to room temperature. The crude product was then partitioned between a saturated aqueous solution of sodium chloride (5 mL) and dichloromelhane (5 mL). The organic layers were combined, dried over sodium sulfate, filtered, evaporated to dryness, and purified on 12 g of silica gel utilizing a gradient of 20-80% ethyl acetate in hexanes to yield I-(benzo[rfir i ,3]dioxoI-5-yI)-W-(2-(2-hydroxypropan-2-yl)- lH-indol-5-yl)cyc!opropanccarboxamide (35 mg, 36%) as a white solid. ESl-MS m/z calc. 378.2, found 379.1 (M+l )+. Retention time of 2.18 minutes. Ή NMR (400 MHz, DMSO-rf6) δ 10.78 (s, 1 I I), 8.39 (s, 1 H), 7.57 (d, J = 1 .7 Hz, I H), 7.1 (d, J = 8.6 Hz, 1 H), 7.03 - 6.90 (m, 4H), 6.12 (dt J = 1.5 Hz, 1 H), 6.03 (s, 211), 5. 18 (s, I H), 1.50 (s, 611), 1.41 - 1.38 (m, 2H), 1.05-0.97 (m. 211).
[00994] Example 113: JV-(2-(l-Amino-2-methylpropan-2-yl)-lH-indol-5-yl)-l. (benzo[d][l^]-dioxol-5-yl)cyclopropanecarboxamide
[00995] Trifluoroacetic acid (0.75 mL) was added to a solution of ierf-butyl 2-(5-( 1 - (benzo[dJ| l ,3]dioxol-5-yl)cyclopropanecarboxamido)- l H-indo]-2-yl)-2-mcthylpropylcarbamatc (77 mg, 0. 16 mmol) in dichloromcthanc (3 mL) and the mixture was stirred at room temperature for 1.5 h. Ί ne mixture was evaporated, dissolved in dichloromelhane, washed with saturated sodium bicarbonate solution, dried over magnesium sulfate and evaporated to dryness to give /V-(2-( l -amino-2-methylpropan-2-yl)- ll-l-indol-5- yl)- l -(benzo[d][ l ,31dioxol-5-yl)cycU)propanc arb«xamide (53 mg, 86%). Ή NMR (400 MHz, CDC13) 8 9,58 (s, 1 H), 7.60 (d, J = 1 .6 Hz, 1 11), 7.18 - 7. 15 (m, 2H), 7.02 - 6.94 (m, 3H), 6.85 (d, J = 7.8 Hz, I H), 6. 14 (d, J = 1.2 Hz, 1 I I), 6.02 (s, 211), 2.84 (s, 2H), 1.68 (dd, J = 3.6, 6.7 Hz, 2H), 1 .32 (s, 6H), 1.08 (dd, J = 3.7, 6.8 Hz, 21-1).
[00996] Example 114: l-iBetizoIdJll^dio ol-S- -A'-tZ-tl^dimeth laminoJ- -rnelh l-propan-2-yl)-lH-indol-5-yl)cyclopropanecarboxamide
[00997] To a solution of N-(2-( 1 -amino-2-methylpropan-2-yl)- 1 H-indol-5-yl)- 1 -(bcnzo[d][l ,3]dioxol-5-yI)cyclopropanccarboxamidc (20 mg, 0.051 mmol) in DMF (I niL) was added potassium carbonate (35 tug, 0.26 mmol) and iodoniethane (7.0 μ1.„ 0.1 1 mmol). The mixture was stirred for 2 h. Water was added and the mixture was extracted with dichloromethane. Combined organic phases were dried over magnesium sulfate, evaporated, coevaporated with toluene (3x) and purified by silica gel chromatography (0-30% i¾OAc in hexane) to give l -(benzo[d]| l ,3|dioxol-5-yl)-/V-(2-( l -(dimethylamino)-2-melhylpropan-2-yl)-1 Il-indol-5-yl)cyclopropanecarboxamide (7 mg, 33%). ]H NMR (400 MHz, CDC13) S 9.74 (s, 111), 7.58 (d, J = 1.9 Hz, 111), 7.20 (d, J = 8.6 Hz, 1 I I), 7.15 (s, 1 H), 7.01 - 6.95 (m, 3H), 6.85 (d, J = 7.9 Hz, III), 6.10 (d, J = 0.9 Hz, 1 I I), 6.02 (s, 2H), 2.43 (s, 211), 2.24 (s, 6H), 1.68 (dd, J = 3.7, 6.7 Hz, 211), 1.33 (s, 6H), 1.08 (dd, J = 3.7, 6.8 Hz, 211).
[00998] Example 1 IS: /V-(2-(l-Acetamido-2-methylpropan-2-yl)-lH-indol-5-yl)-l-(tenzo[d][l^]-dioxol-5-yl)cyclopropanecarboxamide
[00999] To a solution of JV-(2-( 1 -amino-2-methylpropan-2-y1)- 1 H-indol-5-yl)- 1 - (benzo[d][ l,3]dioxol-5-yl)cyciopropanecarboxamide (21 mg, 0.054 mmol) in dichloromethane (1 mL) was added pyridine ( 14 μί, 0.16 mmol) followed by acetic anhydride (6.0 μί, 0.059 mmol). The mixture was stirred for 2 h. Water was added and the mixture was extracted with dichloromethane, evaporated, coevaporated with toluene (3x) and purified by silica gel chromatography (60- 100% ethylacelatc in hexane) to give N-(2-( l -acetamido-2-methylpropan-2-yI)-l H-indol-5-yl)- l -(benzo[d][1 ,3]-dioxol-5-yDcyclopropanecarboxamide ( 17 mg, 73%). Ή NMR (400 MHz, DMSO) 5 10.79 (s, 111), 8.39 (s, 1H), 7.66 (t, J = 6.2 Hz, 1 H), 7.56 (d, J = 1.7 Hz, 1H), 7.18 - 7.14 (m, 111), 7.02 - 6.89 (m, 4H), 6.08 (d, J = 1.5 Hz, I I I), 6.03 (s, 2H), 3.31 (d, J = 6.2 I I/, 2H), 1 .80 (s, 3H), 1.41 - 1.38 (m, 2H), 1.26 (s, 6H), 1.04 - 1.01 (m, 2H).
[001000] Example 116: l -(Benzo[
[001001] l -(Benzok/J[ l,3Jdioxol-5-yl)-A'-(2-(4-cyano-2-methylbutan-2-yl)-lH-indo]-5-yDcyclopropanecarboxamide (83 mg, 0.20 mmol) was dissolved in N,N-dimeLhylformamide (1 niL) containing ammonium chloride (128 mg, 2.41 mmol), sodium azide (156 mg, 2.40 mmol), and a magnetic stir bar. The reaction mixture was heated at 1 10 °C for 40 minutes in a microwave reactor. 'Ihe crude product was filtered and then purified by preparative HPLC using a gradient of 0-99% acetonitrile in water containing 0.05% trifluoroacetic acid to yield l-(benzoli/][l ,3]dioxol-5-yl)-N-(2-(2-mcthy]-4-(lf/-tctrazol-5-yl)butan-2-yl)-l i-indol-5-yl)cyclopropanecarboxamide. ESI-MS m/z cak. 458.2, found 459.2 (M+l )+. Retention time of 1.53 minutes. Ή NMR (400 MHz, CD^CN) 9.23 (s, III), 7.51 - 7.48 (m, 211), 7.19 (d, J = 8.6 Hz, 1H), 7.06 - 7.03 (m, 2H), 6.95 - 6.89 (m, 2H), 6.17 (dd, 7 = 0.7, 2.2 Hz, 1H), 6.02 (s, 2H), 2.61 - 2.57 (m, 211), 2.07 - 2.03 (m, 2H), 1.55- 1.51 (m, 2H), 1.39 (s, 6H), 1.12-1.09 (m, 2H).
[001002] Example 117: i -(Benzo[
[001003] tert-But l 2-(5-(l -(bcnzoM[l ,3]dioxol-5-yl)cyclo-propanecarboxamido)-l H-indol-2-yl)piperidine-l-carboxylate (55 mg, 0.11 mmol) was dissolved in dichloromethane (2.5 mL) containing trifluoroacetic acid (1 niL). The reaction mixture was stirred for 6 h at room temperature. 'Hie crude product was purified by preparative HPLC using a gradient of 0-99% acetonitrile in water containing 0.05% trifluoroacetic acid to yield 1-(benzo[rf][l,3]dioxol-5-yl)-Ar-(2-(piperidin-2-yl)-l -indol-5-yl)cyclopropanecarboxamide. ESI-MS m/z calc. 403.2, found 404.4 (M+l )+. Retention time of 0.95 minutes.
[001004] Example 118: S-/CT-i-Butyl-lH-indol-6-ylamine
[001005] 2-Bromo-4-terf-butyl-pheiiyIamine
[001006] To a solution of 4-im-Butyl-phenylamine (447 g, 3.00 mol) in DMF (500 mL) was added dropwise NBS (531 g, 3.00 mol) in DMF (500 mL) at room temperature. Upon completion, the reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with water, brine, dried over Na2S04 and concentrated. The crude product was directly used in the next step without further purification.
[001007] 2-Bromo-4-fi7t-butyl-5-nitro-phenylamine
[001008] 2-Bromo-4-f
[001009] 4-i£,rt-Butyl-5-nitro-2-triniethylsililnylcthynyl-phenylamine
[001010] To a mixture of 2-bromo-4-ieri-butyl-5-nitro-phenylamine (27.3 g, 100 mmol) in toluene (200 mL) and water ( 100 mL) was added ljN (27.9 mL, 200 mmol), Pd(PPh3)2Cl2 (2.1 1 g, 3.00 mmol), Cul (950 mg, 0.500 mmol) and trimethylsilyl acetylene (21 .2 mL, 1 0 mmol) under a nitrogen atmosphere. The reaction mixture was heated at 70 °C in a sealed pressure flask for 2.5 h., cooled down to room temperature and filtered through a short plug of Celite. The filter cake was washed with RtOAc. '!Tie combined filtrate was washed with 5% NH4OH solution and water, dried over Na SC and concentrated. The crude product was purified by column chromatography (0 - 10 % ethyl acetate/petroleum ether) to provide 4-ί-'r butyl-5-nitro-2-trimethylsilanylethynyl-phen lamίne as a brown viscous liquid (25 g, 81 %).
[001011] 5-terf-Butyl-6-nitro-lH-indole
[001012] To a solution of 4-ίer butyl- -ni-ro-2-trimet lsilan lethynyl-phenylamine (25 g, 86 mmol) in DMF (100 mL) was added Cul (8.2 g, 43 mmol) under a nitrogen atmosphere. The mixture was heated at 1 5 °C in a scaled pressure flask overnight, cooled down to room temperature and filtered through a short plug of Celite. The filter cake was washed with ElOAc. The combined filtrate was washed with water, dried over Na2S04 and concentrated. The crude product was purified by column chromatography (10 - 20 % ethyl aetate hexane) to provide 5-iert-butyl-6-nitro-lH-indole as a yellow solid (13 g, 69 ).
[001013] 5-/err-Butyl-lH-indol-6-ylamine
[001014] Raney Nickel (3 g) was added to 5-ieri-butyl-6-nitro- lH-indole ( 15 g, 67 mmol) in methanol (100 mL). The mixture was stirred under hydrogen (1 atm) at 30 °C for 3 h. The catalyst was filtered off. The filtrate was dried over Na2S04 and concentrated. The crude dark brown viscous oil was purified by column chromatography (10 - 20 % ethyl acetate/petroleum ether) to give 5-½ri-butyl-lH-indol-6-ylamine as a gray solid (11 g, 87 ). 'H MR (300 MHz, DMSO-d6) 5 10.3 (br s, IH), 7.2 (s, 1 H), 6.9 (m, 1H), 6.6 (s, 1H), 6.1 (m, 1H), 4.4 (br s, 2H), 1.3 (s, 9H). a) Ac20, A1C13, CH2C12; b) NaCIO; c) LiAlH4, THF, -78°C; d) SOCl2, CHC13; e) NaCN, DMSO; f) BrCH2CH2Cl, NaOH, Bu NBr, toluene; g) NaOH
[001] Step a: l-(2,3-Dihydro-lH-inden-6-yl)ethanone
[002] A mixture of 2,3-dihydro-lH-indene (100.0 g, 0.85 mol) and acetic anhydride (104.2 g, 1.35 mol) was added drop-wise to a slurry of A1C13 (272.0 g, 2.04 mol) in CH2C12 (1000 ml) at 0 °C over a period of 3h. The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 15 h. Then the reaction mixture was poured into ice water (500 mL) and extracted with ethyl acetate (500 mL x 3). The combined organic layers were washed with brine (500 mL), dried over Na2S04 and evaporated in vacuo. The residue that was purified by column chromatography (petroleum ether : ethyl acetate = 20 : 1 ) to give the product ( 120.0 g, 88%). ]H NMR (400 MHz, CDC13) δ 2.08-2.15 (m, 2H), 2.58 (s, 3H), 2.95 (t, J = 7.2, 4 H), 7.28 (d, / = 8.0, 1 H), 7.75 (d, J = 8.0, 1 H) 7.82 (s, 1 H).
[003] Step b: 2,3-dihydro-lH-indene-5-carboxy]ic acid
[004] To a stirred aqueous sodium hypochlorite solution (2230 ml, 1.80 inmol, 6 %) at 55°C was added l-(2,3-dihydro-l H-inden-6-yl) ethanone (120.0 g,0.75 mol) and the mixture was stirred at 55°C for 2 h. After cooling to room temperature, saturated NallCO.1 solution was added until the solution became clear. The produced precipitate was filtered, washed several times with water and dried to afford the desired product (120.0 g, 99%). Ή NMR (CD I3. 300MHz) δ 2.07-2.17 (m, 2H), 2.96 (t, ./ = 7.5Hz, 4H), 7.30 (d, ,/ =7.8, 1H,), 7. 1 (d, J = 7.8, 1H), 7.96 (s, 1H).
[005] Step c: (2,3-dihydro- 1 H-inden-5-yl)methanol
[006] To a stirred solution of LAH (72.8 g, 1.92 mol) in THF (2.5 L) at 0°C was slowly added 2,3-dihydro- lH-indene-5-carboxylic acid (100.0 g, 0.62 mol). 'the reaction mixture was stirred at 0"C for in. Then the reaction was quenched with H20 (72 ml) and NaOH (68 ml, 20%). The mixture was filtered and the organic layer was dried over Na2S04, evaporated in vacuo and the residue was purified by column chromatography (petroleum ether ; ethyl acetate = 10 : 1 ) to give the desired product (82.0 g, 90%). ]H NMR (CDCla, 300MHz); 6 2.03-2.13 (111, 2H), 2.91 (t, J = 1.5Hz, 4H), 4.64 (s, 2H), 7.1 (d, 7 = 7.5, 1H), 7.18-7.24 (m, 2H).
[007] Step d: 5-(chloromethyl)-2,3-dihydro-lH-indene
[008] Thionyl chloride (120 ml, 1 .65 mol) was added drop-wise to a rapidly stirred mixture of (2,3-dihydro- lH-inden-5-yl)methanol (81.4 g, 0.55 mol) in chloroform (500 ml) at 0°C. After the addition was complete, the resulting mixture was allowed to warm to room temperature and the stirring was continued for an additional 12 h. The chloroform was evaporated under reduced pressure to give a residue, that was purified by column chromatography (petroleum ether : ethyl acetate = 15 : 1 ) to afford 5-(chloromethyl)-2,3-dihydro-lH-indene (90.5 g, 99%). Ή NMR (300 MHz, CDC13) i> 2.06-2.19 (m, 411), 2.93 (t, ./ = 7.5, 4H), 4.54 (s, 2H), 7.15-7.31 (m, 3H).
[009] Step e: 2-(2,3-dihydro-l H-inden-5-yl)acetonitrile
[010] To a stirred solution of 5-(chloromethyl)-2,3-dihydro- 1 H-indene (90.0 g, 0.54 mol) in DMSO (500 ml) was added sodium cyanide (54.0 g, 1.08mol) at 0°C portion wise. The reaction mixture was then stirred at room temperature for 3 hours. The reaction was quenched with water (1000 ml), extracted with ethyl acetate (3 x 250 niL). The combined organic layers were washed with brine, dried over Na2S04 and evaporated in vacuo to afford 2-(2,3-dihydro- lH-inden-5-yl)acetonitrile (82.2 g, 97%), that was used in the next step without further purification.
[011] Step f: l -(2,3-dihydro-l H-inden-6-yl)cyclopropanecarbonitrile
[012] To a stirred solution of 2-(2,3-dihydro- 1 H-inden-5-yl)acetonitrile (50.0 g, 0.32 mol) in toluene (150 mL) was added sodium hydroxide (300 mL, 50 percent in water WAV), l-bromo-2-chlorocthanc (92.6 ml,1.12 mol) and (n-Bu)4NBr (5 g, 15.51 mmol). The mixture was heated at 60 QC overnight. After cooling to room temperature, the reaction mixture was diluted with water (400 mL) and extracted with EtOAc (3 x 200 mL). The combined organic extracts were washed with brine, dried over Na2S04, filtered and concentrated under vacuum and purified by column chromatography (petroleum ether : ethyl acetate = 10 : 1 ) to yield l -(2,3-dihydro-1 H-inden-6-y!)cyclopropaneearbonUrile (9.3 g, 16%). Ή NMR (CDCl3,300MHz) δ 1.35- 1.38 (m, 211), 1 .66- 1.69 (m, 211), 2.05-2.1 (m, 2H), 2.87-294 (iii, 4H), 7.07-7.22 (m,3H).
[013] Step g: l-(2,3-dihydro- lH-inden-6-yl)cyclopropanecarboxylic acid
[014] To a stirred L(2,3-dihydro- l H-inden-6-yl)cyelopropanecarboniu-ile (9.3 g,50.8 mmol) in methanol (40 niL) was added a solution of 150 mL of sodium hydroxide (25% NaOl l w/w in water). The mixture was heated at 100 °C for 8 hours. After cooling lo room temperature, the reaction mixture was poured over ice-water (0 °C), the pH was adjusted to pi 1=4 with hydrogen chloride (1 N) and the mixture was extracted with dichloromelhane (3 x 100 mL). The combined organic layers were dried over a2S04 and evaporated under vacuum. The residue that was purified by column chromatography (petroleum ether : ethyl acetate = 5 : 1) to give L-(2,3-dihydro- lH-inden-6-yDcyclopropanecarboxylic acid (4.8 g,47%). Ή NMR (CDC13, 400 MHz) δ 1.23- 1.26 (m, 211), 1.62-1.65 (m, 2H), 2.03-210 (m, 2H), 2.8 1-2.91 (in, 4H), 7.1 1 -7.21 (m, 3H).
- A mino-2-tert-bu ty 1-1 H-indole-4-carbonitriJe a) KCN, DMSO; b) PcVC, EtOAc
[015] Step a: 2-lert-bulyl-5-nitro- l H-indole-4-carbonitrile
[016] To a solution of 2-teit-butyl-4-nuoro-5-nitro- l H-indole (4.0 g, 17 mmol) in DMSO (30 mL) was added KCN (3.4 g, 51 mmol). The mixture was stirred at 70°C for 3 hours, and poured into water (80 mL) and extracted with ethyl acetate (50 mL x 3). 'lTic combined organic layers were washed with brine, dried over anhydrous NajSO, and concentrated under vacuum. The residue was purified by column chromatography on silica gel (7% EtOAc in petroleum ether) to afford 2-lert-bu.yl-5-nitro- lH-indole-4-carbonitrile (2.2 g, 53%). Ή NMR (DMSO, 300 MHz) δ 12.23 (br s, 1 I I), 8.09 (d, J = 9.0 Hz, 1 H), 7.75 (d, 7 = 9.0 Hz. 1 H), 6.50 (s, 1 H), 1.38 (s, 9 H). MS (ESI) ni/z: 244.2 [M+H4].
[017] Step b: 5-amino-2-tert-butyl- l H-indole-4-carbonitrile
[018] To a solution of 2-lert-butyl-5-nitro- lH-indole-4-carbonitrile (550 mg, 2.3 mmol) in EtOAc (10 mL) was added Rancy Ni (0.1 g) under a nitrogen atmosphere. The mixture was stirred under hydrogen atmosphere ( 1 aim) at room temperature for 1 h. The catalyst was filtered over Celite and the filtrate was evaporated in vacuo to afford 5-amino-2- tert-butyl-1 H-indole-4-carbonitrile (250 mg, 51 %). Ή NMR (DMSO, 300 MHz) δ 10.93 (br s, 1 H), 7.25 (d, J = 8.7 Hz, 1 I I), 6.49 (d, J = 8.7 Hz, 1 H), 5.94 (d, J = 2.1 Hz, 1 H), 5.40 (br s, 2 H), 1.30 (s, 9 H). MS (ESI) ln/z: 214.0 |M+H+j.
N-(2-tert-butyl-4-cyano-lH-indol-5-yl)-l-(2,2-difluorobenzo[d][l^]dioxol-5- y l)cy clopropanecar boxa mide
[019] Step a: N-(2-teri-butyl-4-cyano- 1 H-indoI-5-yl)- I -(2,2-difl u orobenzo[d] [ 1 ,3 J d iox ol- 5 -y l)cy clopropanecarbox amide
[020] 1 -(2,2-difluorobenzo[d][l ,3]dioxol-5-yl)cyclopropanecarbonyl chloride (26 mg, 0.1 mmol) was added to a solution of 5-amino-2-tert-butyl-l H-indole-4-carbonitrile (21 mg, 0.1 mmol) and triethylamine (41.7 ί, 0.3 mmol) in DMF (1 niL). The reaction was stirred at room temperature overnight, then filtered and purified by reverse-phase HPLC to yield the product, N-(2-terl-butyl-4-cyano- lH-indol-5-yl)- l-(2,2-difluorobcnzo[d]| l,31dioxol-5-yl)cyclopropanecarboxamidc. ESI-MS in/z calc. 437.2, found 438.7 (M+l )+. Retention time 2.10 minutes. Ή NMR (400 MHz, DMSO-J6) S 1 1 .48 (s, 1 H), 8.88 (s, IH), 7.52 (d, J = 8.5 Hz, 2H), 7.41 (d, J = 8.3 Hz, 1H), 7.32 (dd, J = 1.5, 8.3 Hz, 1H), 7.03 (d, J = 8.6 Hz, 1H), 6.21 (d, J = 1.8 Hz, 1H), 1.51 - 1.49 (m, 2H), 1.36 (s, 911), 1.18 - 1.16 (m, 2H).
N-(2-tert-butyl-4-cyano-l-(2-hydroxyethy[)-lH-indol-5-yl)-l-(2,2- difluorobenzo[d][l,3]dioxoI-5-y])cyclopropanecarboxamide
[021] Step a: 2-tert-butyl- l-(2-hydroxyethyl)-5-nitro-l H-indole-4-carbonitrile
[022] A mixture of 2-tert-butyl-5-nitro- 1 H-indole-4-carbonitrile (200 mg, 0.82 mmol), 2-iodoethanol (77 ΐ ,, 0.98 mniol), cesium carbonate (534 nig, 1 .64 mmol) and DMF (1.3 mL) was heated to 90 DC overnight. Then more 2-iodoethanol (77 μ\^, 0.98 mmol) was added and the reaction was stirred at 90 °C lor 3 days. The reaction mixture was partitioned between ethyl acetate and water. The aqueous layer was washed with ethyl acetate and then the combined ethyl acetate layers were washed with water (x3) and brine, dried over MgS04 and concentrated. The residue was purified by column chromatography (50 - 100% CH2CI2 - Hexanes) to yield the product as a yellow solid (180 mg, -25% purity by NMR, product co-elutes with the indole starting material). ESI-MS m/z calc. 287. 1 , found 288.5 (M+l )+. Retention time 1.59 minutes. Ή NMR (400 MHz, DMSO-d6) δ 12.23 (s, 1H), 8.14 (d, J = 9.1 Hz, 1 H), 8.02 (d, J = 9.1. Hz, I H), 6.60 (s, 1 H), 5.10 (t, J = 5.5 Hz, 1H), 4.55 (t, J = 6.3 Hz, 2H), 3.78 - 3.73 (m, 2H) and 1.49 (s, 9H) ppm.
[023] Step b: 5-amino-2-terl-butyl- l-(2-hydroxyethyl)-l H-indole-4-carbonitrile
[024] To a solution of 2-tert-butyl- l-(2-hydroxyethyl)-5-nitro-lH-indole-4-carbonitrile ( 180 mg, 0.63 mmol) in ethanol (6 mL) under N2 atmosphere was added Pd-C (5% wl, 18 mg). The reaction was flushed with N2 (g) and then with H2 (g) and stirred under H2 (atm) at room temperature for 1.5 hours. The reaction was filtered over Cclitc and concentrated to yield the product (150 mg, 93 %). ESI-MS tn/z calc. 257.2, found 258.5 (M+ l )+. Retention time 1 .26 minutes.
[025] Step c: N-(2-tert-butyl-4-cyano- l -(2-hydroxyethyl)- lII-indol-5-yl)- I -(2,2-difluorobenzo[d] [ 1 ,3]dioxol-5-yl)cyclopropanecarboxamide
[026] l -(2,2-dilluorobenzo[d][l,3]dioxol-5-yl)cyclopropanecarbonyl chloride ( 1 6 mg, 0.75 mmol) was added to a solution of 5-amino-2-tert-butyl- l-(2-hydroxyethyI)-lH-indole-4-carbonitrile (150 mg, 0.58 mmol) and triethylamine (242 μί, 1.74 mmol) in dichloromethane (2 mL). The reaction was stirred at room temperature overnight. The reaction mixture was diluted with dichloromethane and extracted with IN HC1 solution (x2), saturated NaHCOa solution (x2), brine, dried over MgS04, filtered and concentrated. ITie residue was dissolved in DMSO and purified by reverse-phase HPLC to yield the product, N-(2-terl-butyl-4-cyano- l-(2-hydroxyethyl)- lII-indol-5-yl)-l-(2,2-difluorobenzo[d][l,3]dioxol-5-yl)cyclopropanecarboxamide. ESI-MS m/z calc. 481 .2, found 482.5 (M+l)+. Retention time 1.99 minutes. Ή NMR (400 MHz, DMSO-rf6) δ 8.93 (s, IH), 7.71 (d, J = 8.8 Hz, II I), 7.51 (s, IH), 7.42 (d, J = 8.3 Hz, III), 7.33 (d, J = 1.6 Hz, III), 7.08 (d, J = 8.8 Hz, I H), 6.28 (s, 111), 5.05 (t, J = 5.6 Hz, I H), 4.42 (t, J = 6.8 Hz, 2H), 3.70 - 3.65 (m, 2H), 1.51 - 1.48 (m, 2H), 1.44 (s, 9H), 1.19 - 1. 16 (m, 2H). 2-(2-iert-butyl-5-(l-(2,2-difluorobenzo[d]tl,3]dioxol-S- yl)cyclopropanecarboxamido)-6-fluoro-lH-indol-l-yl)-N,N,N- trimethyiethanaminium chloride
[027] Step a: fert-Butyl 2-(2-/eri-butyl-5-(l-(2,2-difluorobenzo|;d][l,3]dioxol-5-yl)cyclopropanecarboxaniido)-6-fluoro- ll-l-indol- 1 -yl)et ylcarbamate
[028] To l -(2,2-difluorobenzo[d][l ,3]dioxol-5-yl)cyclopropaneearboxylic acid (90.14 mg, 0.3722 mmol) in thionyl chloride (81 .28 μί, 1.117 mmol) was added N,N -dimethyl formamidc (8.204 μί, 0.1064 mmol). The reaction mixture was stirred at room temperature for 30 minutes before excess thionyl chloride and N,N -dimethyl formaniide were removed in vacuo to yield the acid chloride. The acid chloride was then dissolved in dichloromethane ( 1.5 mL) and added slowly to a solution of rert-butyl 2-(5-amino-2-/ert-bulyl-6-lluoro-lH-indol- l-yl)el ylcarbamate ( 156.1 mg, 0.4467 mmol) and triethylamine (155.6 μί, 1.1 17 mmol) in dichloromethane ( 1.5 mL). T¾e resulting reaction mixture was stirred at room temperature for 21 hours. 1 Tie reaction mixture was diluted with dichloromethane (5 mL) and washed with IN aqueous HCl (5 niL) and a saturated aqueous NaHCOa solution (5 mL). The organic layer was dried over Na2S04, filtered and evaporated under reduced pressure. The crude product was purified by column chromatography on silica gel (0-30% ethyl acetate in hexane) to yield ½rt-butyl 2-(2-ieri-butyl-5-(l-(2,2-difluorobenzo|d][ l ,3]dioxol-5-yl)cyclopropanecarboxamido)-6-fluoro-lH-indoI- l -yl)ethylcarbamate as a white solid ( 140 mg, 66%). ESI-MS tn z calc. 573.2, found 574.7 (M+ l)+. Retention time 2.4] minutes. IH NMR (400.0 MHz, DMSO) d 8.35 (s, 1H), 7.53 (s, 1H), 7.44 - 7.41 (m, 2H), 7.34 - 7.29 (m, 2H), 7.13 - 7.10 (m, 1H), 6.17 (s, 1H), 4.24 -4.20 (m, 2H), 3.20 - 3.17 (m, 2H), 1.48- 1.45 (m, 2H), 1.41 (s, 18H) and 1.15- 1. 12 (m, 2H) ppm.
[029] Step b: N-(l-(2-aminoethyl)-2-ieri-butyl-6-fluoro-lH-indol-5-yl)- l-(2,2- dinu()rubenzo[propanecarb()xaniide
[030] To a solution of /err-but l 2-(2-½r/-butyl-5-(1 -(2,2-dilluoiObenzo|d)| l,3|dioxol-5-yl)cyclopropanecarboxaniido)-6-nuoro- I H-indo]- l -yl)elhylcarbamate (137.5 mg, 0.24 mmol) in dichloroinelhane ( 1.8 ml.) was added tiifluoroacelic acid (444 μί, 5.8 mmol) and the mixture was stirred at room temperature for 1 hour. The reaction was diluted with dichloromethane and washed with saturated aqueous Nal lC03 solution (3 mL) and brine (3 mL). The organic layer was dried over Na2S04, filtered and evaporated under reduced pressure. 'IThe crude product was purified by column chromatography on silica gel (0- 10% methanol in dichloromethane) to yield N-( l -(2-aminoethyl)-2-ieri-butyl-6-fluoro- lH-indol-5-yl)- l-(2,2-difluorobenzo|d][ 1 ,3]dioxol-5-yl)cyclopropanecarboxamide as a while solid (93.7 mg, 82%). BSI-MS m z calc. 473.19, found 474.5 (M+! )+. Retention time 1.61 minutes.
[031] Step c: 2-(2-/er/-butyt-5-( l-(2,2-difluorobenzo[d]tl ,3]dioxol-5-yl)eyclopropanecarboxamido)-6-fluoro- 1 H-indol-1 -yl)-N,N,N-trimethylelhanan)iniuin chloride
[032] To a clear solution of N-( l-(2-aminoethyl)-2-ieri-butyl-6-fluoro- l H-indol-5-yl)- l -(2,2-difluorobenzo[d]| l ,3jdioxol-5-yl)cyclopropanecarboxamide (50 mg, 0.1056 mmol) in N,N -dimethyl formamide ( 1 mL), methyl iodide (336.8 mg, 147.7 μΒ, 2.37 mmol) and triethylamine (106.9 mg, 147.2 L, 1 .05 mmol) were added and the mixture was heated at 80 °C for 2 hours. Ύ α crude product was purified by reverse phase preparative HPLC 22 mg of this product were dissolved in 1.25 HCl in methanol (112 μΐ., 0.14 mmol) and heated at 60 °C for 1 hour. The reaction was cooled to room temperature. The product was first dried and then dissolved in dichloromethane and dried again. This procedure was repealed four times to yield 2-(2-ieri-butyl-5-( l-(2,2-difluorobenzoldj| l,3]dioxol-5-yl)cyclopropanecarboxamido)-6-fluoro- lH-indol-l-yl)-N,N,N-trimethylelhanaininium chloride. ES1- S m j calc. 516.25, found 516.7 (M+ l }+. Retention time 1.69 minutes. 1 H NMR (400.0 MHz, DMSO) d 8.43 (s, 1 H), 7.53 (s, 111), 7.45 - 7.41 (m, 211), 7.36 - 7.31 (m, 2H), 6.27 (s, 1 H), 4.74 - 4.70 (m, 2H), 3.57 - 3.53 (m, 2H), 3.29 (s, 9H), 1.48 - 1.42 (m, 1 1 H), and 1.15 (dd, J = 3.9, 6.8 Hz, 2H) ppm. 2-(4-(Tert-butyldimethylsilyloxy)-2-methylbutan-2-yl)-6-tluoro-5-nitro-lH- indole
[033] Step a: 3-fluoro-4-nitroaniline 1034] A mixture of N-(3-fluoro-4-nitro-phenyl)-2, 2-dimet yl-propionamide (87.0 g, 0.36 mol) in CH2C12 (400 niL) and 6N hydrochloric acid (800 mL) was heated to reflux for 2 hours. 'l"he reaction mixture was cooled to room temperature. 'Hie reaction mixture was diluted with 1000 mL of ethyl acetate and potassium carbonate (500.0 g) was added portion wise. The aqueous solution was separated and the organic layer was washed with brine and dried over anhydrous Na2S04. The solvent was removed by evaporation under reduced pressure; the residue was purified by column chromatography on silica ge) (petroleum ether / ethyl acetate 30: 1) to afford 3-fluoro-4-nitroaniline (56.0 g, 99 %). ]H NMR (300 MHz, CDC13) 6 8.07 (t, J = 8.7 Hz, 1 11), 7.86 (dd, J= 2.1 , 13.2 Hz 1 H), 7.59 (brs, 2 H), 7.22 (s, 1 H).
[035] Step b: 2-bromo-5-fluoro-4-nitroaniline To a solution of 3-fluoro-4-nitroaniline (56 g, 0.36 mol) in acetic acid (500 mL) was added drop-wise bromine (17.7 mL, 0.36 mol) over 1 hour. The reaction mixture was stirred for 1 hour at 0-5 °C in an ice bath. The reaction mixture was basified with saturated Na2C03 and extracted with ethyl acetate (200 niL x 3). 'Hie combined organic layers were washed with brine, dried over anhydrous Na2S04, filtered and concentrated under reduced pressure to yield a residue that was purified by column chromatography on silica gel (petroleum ether / ethyl acetate 10 : 1) to give the 2-bromo-5-fluoro-4-nitroanilinc ( 45.6 g, 84 % ) as a yellow solid. Ή NMR (400 MHz, CDC13) δ 8.29 (d, J = 7.6 Hz, 1 H), 653 (d, J = 12.4 Hz, 1 H), 4.94 (br s, 2 H).
[036] Step c: ethyl 5-(2-amino-4-fluoro-5-nitrophcnyl)-3,3-dime-hylpenl-4-ynoaie
[037] To a solution of 2-bromo-5-f]uoro-4-nitroaniline (45.7 g, 0.19 mol) and ethyl 3,3-dimethylpent-4-ynoate (88.3 g, 0.57 mol) in Et3N (700 mL) was added Pd(PPh3)2Cl2 ( 13.8 g, 0.02 mol) and Cut (3.6 g, 0.02 mol) under N2. The reaction mixture was stirred at 70°C for 8 hours. The reaction mixture was diluted with 500 mL of ethyl acetate and 1500 mL of water. The organic layer was separated and the aqueous phase was extracted with ethyl acetate (500 mLx3), the combined organic layers were washed with brine and dried over anhydrous Na2S04, filtered and evaporated under reduced pressure and the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate 1 U: 1 ) to give ethyl-5-(2-amino-4-fiuoro-5- nitrophenyl)-3,3-dimethylpent-4-ynoate (34.5 g, 57 %). Ή NMR (300 MHz, CDCIj) δ 8.05 (d, J = 8. l Hz, 1 H), 6.36 (d, J = 13.2 Hz, 1 H), 5.60 (brs, 2 H), 4. 16 (q, J = 7.2 Hz, 2 H), 2.51 (s, 2 H), 1.40 (s, 6 H), 1.28 (t, J = 7.2 Hz, 3 II).
[038] Step d: ethyl 3-(6-fluoro-5-nitro- lH-indol-2-yl)-3-methylbutanoate
[039] To a mixture of ethyl 5-(2-amino-4-fluoro-5-nitropheny3)-3, 3- dimethylpent-4-ynoate (34.5 g, 0.1 1 mol) and PdCI2 ( 10.4 g, 58.6 nniol) in CH3CN (350 mL) was heated to reflux for 1.5 hours. The reaction mixture was cooled down to room temperature. Ethyl acetate (300 mL) was added, the precipitate was filtered off and washed with methanol. The filtrate was concentrated under reduced pressure and the residue was puri fied by column chromatography on silica gel (petroleum ether / ethyl acetate 40: 1 ) to give ethyl 3-(6-fluoro-5-nitro-i H-indol-2-yl)-3-niethylbutanoate (34.0 g, 98 ) as a deep yellow solid. Ή NMR (300 MHz, C C13) 6 10.1 1 (brs, 1 H), 8.30 (d, J = 7.2 Hz, 1 H), 7.14 (d, ./ = 1 1 .7 Hz, 1 H), 6.35 (d, ./ = 1.5 Hz, 1 H), 4.17 (q, ./ = 7.2 Hz, 2 H), 2.69 (s, 2 H), 1.51 (s, 6 11), 1.25 (t, 7 = 7.2 Hz, 3 I I).
[040] Step e: 3-(6-fluoro-5-nitiO- lH-indo]-2-yl)-3-methylbutan- l -ol
[041] To a solution of ethyl 3-(6-fluoro-5-nitro-lH-indol-2-yl)-3- methyibutanoate (34 g, 0.1 1 mol) in dry CH2C12 (400 mL) was added drop-wise DIBAL-H (283.4 mL, 0.27 mol) over 2 hours at -78°C. The reaction mixture was stirred for 10 hours at -78°C and then quenched by adding water (200 mL). The precipitate was filtered off and washed with methanol. The filtrate was extracted with CH2C12 (2(X) mLx3), the combined organic layers were washed with brine, dried over anhydrous Na2S04 and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate 50: 1) to give 3-(6-fluoro-5-nitro-l H-indol-2-yl)-3- methylbutan-l -ol (6.6 g, 22 ). Ή NMR (400 MHz, CDC13) δ 9.35 (brs, 1 II), 8.30 (d, J = 7.6 Hz, 1 H), 7.1 1 (d, J = 12.0 Hz, 1 H), 6.35 (d, J = 1.2 Hz, 1 H), 3.74 (t, J = 6.4 Hz, 2 H), 1.9 (1, 7 = 6.4 Hz, 2 H), 1.4 (s, 6 H).
[042] Step f: 2-(4-(iert-butyldiniethylsilyloxy)-2-methylbulan-2-yl)-6-fluoro-5-nitro-lH-indole
[043] To a solution of 3-(6-fluoro-5-nitro- lH-indoI-2-yl)-3-methylbutan-l-o] (6.6 g, 25 mmol) in CH2CI2 (80 mL) was added TBSCl (3.7 g, 25 nniol) and imidazole (4.2 g, 62 nmol) at 0°C. The reaction mixture was stirred at room temperature for 12 hours. The precipitate was filtered off and washed with the methanol. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate 10: 1) to give the desired product as a brown solid (5.0 g, 53 %). Ή N R (300 MHz, C C13) 8 9.80 (brs, I H), 8.30 (d, J = 7.2 ΙΙζ, Ι H), 7.05 (d, J = I i .7 Hz, 1 H), 6.33 (t, / = 1.2 Hz, 1 11), 3.7 (t, J = 6.0 Hz, 2 I I), 1. 1 (l, J = 6.0 Hz, 2 I I), 1.42 (s , 6 H), 0.94 (s , 9 H), 0. 12 (s , 6 I I). MS (ESI) m/z (M+H+): 381 .1.
Benzyl 2,2-dimethylbut-3-ynoate
[044] Step a: methyl 2,2-dimeth l-3-oxobulanoate
[045] To a suspension of NaH (28.5 g, 0.718 mol, 60%) in THF (270 mL) was added dropwise a solution of 3-oxo-butyrie acid methyl ester (78.6 g, 0.677 mol) in THF (70 mL) at 0 °C. ITie mixture was stirred for 0.5 hours al 0 °C Mcl (99.0 g, 0.698 mol) was added dropwise at 0 °C. The resultant mixture was warmed to room temperature and stirred for I hour. NaH (28.5 g, 0.718 mol, 60%) was added in portions al 0 UC and the resulting mixture was continued to stir for 0.5 h at 0 °C. Mcl (99.0 g, 0.698 mol) was then added dropwise al 0 °C. Plhe reaction mixture was warmed to room temperature and stirred overnight. The mixture was poured into ice water. The organic layer was separated. The aqueous phase was extracted with EtOAc (300 mL x 3). The combined organic layers were dried and evaporated under reduced pressure to give methyl 2,2-dimelhyl-3-oxobutanoate (52 g, 53%), which was used directly in the nexl step.
[046] Step b: methyl 3-chloro-2(2-dimcthylbut-3-cnoaLe
[047] To a suspention of PC15 (161 g, 0.772 mol) in dichloromcthanc (600 mL) was added dropwise methyl 2,2-dimelhy]-3-oxobutanoate (52 g, 0.361 mol, crude from last step) at 0 °C, followed by the addition of approximately 20 drops of dry DMF. The mixture was heated at reflux overnight. After cooling, the reaction mixture was slowly poured into ice water. The organic layer was separated and the aqueous phase was extracted with dichloromethane (300 mL x 3). The combined organic layers were washed with saturated aqueous Nal lCOj solution and dried over anhydrous Na2S04. The solvent was evaporated to give the product, methyl 3-chloro-2,2-dimethylbut-3-enoate which was used without further puri fication (47 g, 82%).
[048] Step c: 3-ch1oro-2,2-dimethylbut-3-enotc acid
[049] Λ mixture of methyl 3-chloro-2,2-dimethylbut-3-enoate (42.0 g, 0.26 mol) and NaOH ( 12.4 g, 0.31 mol) in water (300 ml was heated at reflux overnight. Alter cooling, the reaction mixture was extracted with ether, The organic layer contained 20g of methyl 3-chloro-2,2-dimethylbut-3-enoate (48 % recovered). The aqueous layer was acidified with cold 20% HC1 solution and was extracted with ether (250 mL x 3). The combined organic layers were dried and evaporated under reduced pressure to give 3-chloro-2,2-dirncihylbut-3-enoic acid ( 17 g, 44 %), which was used directly in the next step.
[050] Step d: 2,2-dimethylbut-3-ynoic acid
[051] To a three-neck flask (500 mL) was added NaNII2 ( 17.8 g, 0.458 mmol, pellets) and DMSO (50 mL). The mixture was stirred at room temperature until no more NH3 (g) was given off. A solution of 3-chloro-2,2-diniethylbut-3-enoic acid ( 17.0 g, 1 14 mmol) in DMSO (50 mL) was added dropwise at 0 °C. The mixture was warmed and stirred at 50 °C for 5 hours, then stirred at room temperature overnight. 1 Tie mixture was poured into cold 20% HC1 solution, and then extracted uVee times with ether. The ether extracts were dried over anhydrous Na2S04 and concentrated to give a 6: 1 ratio of starting material and alkyne product. The residue was re-dried using ether and Na2SU and re-subjected to the reaction conditions above. The reaction mixture was worked up in the same manner to provide 2,2-dimethylbut-3-ynoic acid (12.0 g, 94 %).
[052] benzyl 2,2-dimethylbut-3-ynoate
[053] To a stirred solution of 2,2-dimethylbut-3-ynoic acid (87.7 g, 0.782 mmol) and benzyl alcohol (1 4.6 g, 0.938 mol) in dichloromethane (800 mL) was added DCC ( 193.5 g, 0.938 mmol) at -20 °C. The reaction mixture was stirred at room temperature overnight and then the solvent was evaporated in vacuo. The residue was purified by chromatography on silica gel (2% ethyl acetate in petroleum ether as eluant) to afford benzyl 2,2-dimethylbut-3-ynoate (100 g, 59 % yield). lH NMR (CDC13.400 MHz) 5 7.37-7.36 (m, 5 H), 5.19 (s, 2 H), 2.28 (s, 1 H), 1.52 (s, 6 H). 2-(l-(Tert-butyldimethylsily!oxy)-2-methylpropan-2-y|)-6-fluoro-5-nitro-lH- indole
[054] Step a: benzyl 4-(2-amino-4-fluoro-5-nitrophenyl)-2,2-dimethylbut-3-ynoate
[055] To a solution of 2-bromo-5-nuoro-4-nitroaniline (23.0 g, 0.1 mol) in Et3N (250 mL) was added benzoic 2,2-diniethy]bul-3-ynoic anhydride (59.0 g, 0.29 mol), Cul ( 1.85 g) and Pd(PPh3)2Cl2 (2.3 g) at room temperature. The mixture was stirred at 80°C overnight. After cooling to room temperature, the reaction was quenched with water and the aqueous layer was extracted with ethyl acetate ( LOO mL x 3). The combined organic layer was dried over anhydrous Na2SO.i, the solvent was evaporated in vacuo. The residue was purified by chromatography on silica gel ( 10% ethyl acetate in petroleum ether) to give benzyl 4-(2-amino-4-lluoro-5-nitrophenyl)-2,2-dimethylbut-3-ynoate (20.0 g, 56%). ]l l NMR (400 MHz, CDC13) 8.05 (d, J = 8.4 Hz, 1 H), 7.39-7.38 (m, 5 H), 6.33 (d, J = 13.2 Hz, 1 H), 5.20 (s, 2 H), 4.89 (br s, 2 H), 1.61 (s, 6 H).
[056] Step b: benzyl 2-(6-fluoro-5-nitro- 1 H-indol-2-yl)-2-melhylpropanoate
[057] To a solution of benzyl 4-(2-aniino-4-nuoro-5-nitrophenyl)-2,2-dimethylbut-3-ynoate (20.0 g, 56 mmol) in acetonitrile ( 100 mL) was added PdCl2 (5.0 g, 28 mmol) at room temperature. The mixture was stirred at 80"C overnight. The mixture was filtered off and the solvent was evaporated in vacuo, the residue was purified by chromatography on silica gel (10% EtOAc in petroleum ether) to give benzyl 2-(6-fluoro-5-nitro- lH-indol-2-yl)-2-methylpropanoate (18.0 g, 90%). Ή NMR (300 MHz, CDC1 ) 8.96 (br s, 1 H), 8.33 (d, / = 7.2 Hz, 1 H) 7.35-7.28 (m, 5 H) 7.08 (d, / = 1 1.7 Hz, 1 H), 6.47 (s, 1 H), 5.18 (s, 2 H) 1-69 (s, 6 I I).
[058] Step c: 2-(6-fluoro-5-nilro- lH-indol-2-yl)-2-methylpropan- l -ol To a solution of benzyl 2-(6-fluoro-5-nitto- l H-indoI-2-yl)-2-methylpropanoate (18.0 g, 0.05 mol) in CH2C12 (100 mL) was added DIBAL-H (12 mL) at -78°C. The mixture was stirred for 1 h al that temperature and was warmed to room lemperalure. The reaction was quenched with water and the aqueous layer was extracted with lilOAc ( 100 mL x 3). 'Ihe combined organic layers were dried over anhydrous Na2S0 , the solvent was evaporated in vacuo. The residue was purified by chromatography on silica gel ( 10% EtOAc in petroleum ether) to give 2-(6-nuoro-5-nitro- l ll-indol-2-yl)-2-methylpropan- l -ol (10.0 g, 77%). !H NMR (300 MHz, CDCb) 9.37 (s, 1 H), 8.32 (d, J = 7.2 Hz, 1 H), 7.1 1 (d, J = 1 ! .7 Hz, 1 H), 6.36 (s, 1 H), 3.73 (d, 7 = 5.1 Hz 2 H), 1 .97 (t, J = 5.1 Hz, 1 H), 1.39 (s, 6 H).
[059] Step d: 2-(l -(tert-butyldimethylsilyloxy)-2-methylpropan-2-yl)-6-fluoro-5-nitro--indole
[060] To a stirred solution of 2-(6-fluoro-5-nitro- 1 H-indol-2-yl)-2-mcthylpropan- 1 -ol (lO.Og) in CH2CI2 was added TBSC1 (8.9 g), imidazole (8.1 g, 0.12 mol) al room temperature. Ί Tie mixture was stirred overnight. The solvent was evaporated in vacuo and the residue was purified by chromatography on silica gel (10% EtOAc in petroleum ether) to give 2-( l-(tert-butyldiniemylsilyloxy)-2-memylpropan-2-yl)-6-nuoro-5-nitro- l H-indole (5.3 g, 38 %). Ή NMR (300 MHz, CDCb) 9.51 (s, 1 II), 8.31 (d, ./ = 7.5 Hz, 1 1 1), 7.02 (d, J = 1 1.7 Hz, 1 H), 6.32 (s, 1 H), 3.63 (s, 2 H), 1.35 (s, 6 H), 0.99 (s, 9 H), 0.1 1 (s, 6 H). 6-fluoro-l,l-dimethyl-7-nitro-2,3-dihydro-lH-pyrrolo[l,2-a]indole, (R)-3- (l-((2,2-dimethyl-ls3-dioxolan-4-yl)methyl)-6-fluoro-5-nitro-lH-indol-2- y -3-methylbutan-l-ol, 2-(4-(((R)-2,2-dimethyl-l,3-dioxoIan-4- y])methoxy)-2-methylbutan-2-yl)-l-(((R)-2,2-dimethyl-l,3-dioxolan-4- yI)methyl)-6-fluoro-5-nitro-lH-indole, 3-(6-fluoro-5-nitro-lH-indol-2-yl)-3- methylbutan-1 -ol and (R)-2-(4-((2,2-dimethyl-l,3-dioxolan-4-yl)methoxy)- 2-methylbutan-2-yl)-6-fluoro-5-nitro-lH-indole
[061] Step a: 6-fluoro- i , Ndimethyl-7-nitro-2,3-dihydro-lH-pyrrolo[ l ,2-alindole, (R)-3-( l -((2,2-dimethyl-l ,3-dioxolan-4-yl)methyl)-6-nuoro-5-nitro-l H-indol-^ methylbutan-1 -ol, 2-(4-(((R)-2,2-dimethyl-l,3-dioxolan-4-yl)methoxy)-2-mclhylbutan-2-yl)- l -(((R)-2,2-dime l- l ,3-dioxolan-4-yl)niethyl)-6-fluoro-5-nitro- lH-indole, 3-(6-fiuoro-5-nitro- 1 H-indol-2-yl)-3-methylbuian- l-ol and (R)-2-(4-((2,2-dimethyl- 1 ,3-dioxolan-4-yl)i]iethoxy)-2-iiielhylbulan-2-yl)-6-lluoro-5-nilro- lH-indole
[062] To a solution of 2-(4-(tert-butyldimethylsilyloxy)-2-methylbutan-2-yl)-6-fluoro-5-nitro- l H-indole ( 1.9 g, 5.0 mmol) and (S)-(2,2-dimethyl- l ,3-dioxolan-4-yl)methyl 4-niethylbenzenesulfonate (2.86 g, 10.0 mmol) in DMF ( 10 ml ,) was added Cs2C03 (4.88 g, 15.0 mmol). The mixture was heated at 90 "C for 24 hours. The reaction was partitioned between ethyl acetate and water. The aqueous layer was extracted with ethyl acetate and the combined organic layers were washed with brine and dried over gS0 . After the removal of solvent, the residue was purified by column chromatography ( 10-50% ethyl acetate -hexane) to afford 6-fluo )-l , l -dime(hy]-7-nitro-2,3-dihydro- l H-pyrrolo| l ,2-a]indole (600 mg, 48%). ESI-MS w¾ calc. 248.1 , found 249.2 ( + 1 )+. Retention time 2.00 minutes; 2-(4-(((R)-2,2 -dimethyl- 1 ,3-dioxolan-4-yl)methoxy)-2-methylbutan-2-yl)- i -(((R)-2,2-dimethyl-l ,3-dioxolan-4-yl)mcthyl)-6-fluoro-5-nitro- l H-indolc (270 mg, containing some (R)-2-(4-((2,2-dmiemy 1- 1 ,3-dioxolan-4-yl)me^ ESI-MS m z calc. 494.2 and 380.2, found 495.4 and 381.4 (M+ 1 f. Retention time 2.12 and 1.92 minutes; (R)-3-( l-((2,2-dimethyl-l ,3-dioxolan-4-yl)mcthyl)-6-iluoro-5-nitro- 1 l-I-indol-2-yl)-3-methyIbutan- l-ol ( 1.0 g, containing some 3-(6-fluoro-5-nitro- I H-indol-2-yl)-3-methylbutan- No!). ESI-MS m/z calc. 380.2 and 266.1 , found 381.2 and 267.2 (M+l )+.
Retention time 1.74 and 1 .48 minutes.
(R)-2-(l-((2,2-dimethyl-l,3-dioxolan-4-yl)methyl)-6-fluoro-5-nitro-lH-indol- 2-yl)-2-methylpropan-l-ol and 3-(6-fluoro-5-nitro-lH-indol-2-yl)-3- methylbutan-l-ol
[063] A mixture containing (R)-2-( 1 -((2,2-dimethyl- 1 ,3-dioxolan-4-y])meihyl)-6-fluoro-5-nitro- lH-indol-2-yl)-2-methylpropan- J -ol and 3-(6-fluoro-5-nitro- I H-indol-2-yl)-3-methylbuian-l -ol was obtained following the procedure shown above starting from 2-(l -(tert-butyldimethylsilyloxy)-2-methylpropan-2-yl)-6-fluoro-5-nitro- lH-tndole. (R)-2-(l -((2,2-dimethyl-l,3-dioxolan-4-yl)methyl)-6-fluoro-5-nitro-l H-indol-2-yl)-2-methylpropan-l -ol, ESI-MS m/z calc. 366,2, found 367.2 (M+l )+. Retention time 1.71 minutes; 3-(6-fluoro-5-nitro-ll-l-indol-2-yl)-3-methylbulan- l -ol, ESI-MS m/z calc. 252.1 , found 253.4 (M+ l)+. Retention time 1.42 minutes. l-(2,2-dinuorobenzo[d][l,3]dioxol-5-yl)-N-(6-fluoro-l,l-dimethyl-2,3-dihydro- lH-pyrrolo[l,2-a]indol-7-yl)cyclopropanecarboxamide
[064] Step a: 6-fluoi\)-l , l -dinicthyl-2,3-dihydm- I H-pyrrok>[ l ,2-a]ind )I-7-un]ine
[065] To a solution of 6-fluoro- l , l -dimethyl-7-nitro-2,3-dihydro-l I I-pyrroloL l ,2-aJindole (600 mg, 2.4 mmol) in ethanol (15 niL) was added ammonium formate (600 mg, 9.5 mmol) and Pd/C ( 10%, 129 mg, 0.12 mmol). The mixture was refluxed for 10 min. The Pd catalyst was removed via filtration through Celite and washed with ethanol. The filtrate was concentrated and purified by column chromatography (20-40% ethyl acetate-hcxancs) to provide 6-fluoro- l , l-dimcthyl-2,3-dihydro- l H-pyrrolo( l ,2-ajindol-7-aminc (260 mg, 49 %). ESl- S m/z calc. 218.1 , found 219.2 (M+ l f . Retention time 1.01 minutes.
[066] Step b: l -(2,2-difluorobenzo[dj[l,3jdioxol-5-yl)-N-(6-fluoro- l 1 l -dimethy]-2,3-dihydro- l H-pyrrolo[ l ,2-a]indol-7-y])cyclopropanecarboxamide
[067] To a mixture of 1 -(2,2-difluorobenzo[d |[1 ,3]dioxol-5-yl)cyclopropanecarboxylic acid (346 mg, 1.4 mmol), 6-fiuoro- l ,l -dimethyl-2,3-dihydfo- l H-pyrrolol 1 ,2-a]indol-7-amine (260 mg, 1 .2 mmol) and HATU (543 mg, 1.4 mmol) in DM!7 (5 niL) was added triethylamine (0.40 mL, 2.9 mmol). ITie reaction was stirred at room temperature overnight and then partitioned between ethyl acetate and water. The aqueous layer was extracted with ethyl acetate and the combined organic layers were washed with brine and dried over MgS04. After the removal of solvent, the residue was purified by column chromatography (10-20% ethyl acetate - hexanes) to afford 1 -(2,2-difluorobenzo| d] 1 1 ,3]dioxol-5-yl)-N-(6-fluoro- 1 , l-dimethyl-2,3-dihydro- 1 M-pyrrolo| 1 ,2-a|indol-7-yl)cycloprapanecarboxamide (342 mg, 65 %). RSI-MS tn z calc. 442.2, found 443.5 (M+ l f. Retention time 2.30 minutes. Ή NMR (400 MHz, DMSO-i/6) δ 8.20 (d, J = 7.6 Hz, 1H), 7.30 - 7.25 (m, 311), 7.20 (m, 1H), 7.12 (d, J = 8.2 Hz, 1H), 6.84 (d, J = 1 1.1 Hz, 1 H), 6.01 (d, J = 0.5 Hz, 111), 3.98 (t, J = 6.8 Hz, 2H), 2.37 (t, J = 6.8 Hz, 2H), 1.75 (dd, J = 3.8, 6.9 Hz, 2H), 1.37 (s, 6H) and 1 . 14 (dd, J = 3.9, 6.9 Hz, 2H) ppm.
(R)-l-(2,2-difluorobenzo[d][l,3]dioxol-5-yl)-N-(l-(2,3-dihydroxypropyl)-6- fluoro-2-(4-hydroxy-2-methylbutan-2-yl)-lH-indol-5- y 1 )cy clop ro panecarboxa mi de
[068] Step a: (R)-3-(5-umino- 1 -({2,2-dimethyl- 1 ,3-dioxolan-4-yl)methyl)-6-fluon> 1 H-indol-2-yl)-3-methylbutan- 1 -ol
[069] To a solution of (R)-3-( 1 -((2,2-dimethyl- 1 ,3-dioxolan-4-yl)melhyl)-6-fluoro-5-nitro- I H-indol-2-yt)-3-methylbutan- i -ol containing some 3-(6-fluoro-5-nitro- l H-indol-2-yl)-3-methylbutan- l -ol (500 mg, 1.3 mmol) in ethanol ( 10 mL) was added ammonium formate (500 mg, 7.9 mmol) and Pd/C (10%, 139 mg, 0.13 mmol). The mixture was refluxcd for 5 min. The Pd catalyst was removed via filtration through Ce!iie and washed with ethanol. Ihe filtrate was evaporated to dryness and purified by column chromatography (30-50% ethyl acetate-hexanes) to provide (R)-3-(5-amino- l -((2,2-dimethyl-l ,3-dioxolan-4-yl)methyl)-6-fluoro- l H-indol-2-yl)-3-methylbutan- l-ol (220 mg, 48 %, contains some 3-(5-amino-6-fluoro-lI l-indol-2-yl)-3-methylbulan- l-ol). ESI-MS tnJz calc. 350.2 found 351.4 (M+l )+. Retention time 0.94 minutes.
[070] Step b: (R)- l -(2,2-difluorobcnzo[d]l 1 ,3]dioxol-5-yl)-N-( l -((2,2-dimethyl- 1 ,3-dioxolan-4-yl)nielhyl)-6-fluoro-2-(4-hydroxy-2-methylbutan-2-yl)- l H'indol-5-yl)cyclopropanecarboxamide
[071] To a mixture of l -(2,2-difluoroberizo|d][ l ,3 |dioxoL5-yl)cyclopropanecarboxylic acid ( 183 mg, 0.75 mmol), (R)-3-(5-amino- 1 -((2,2-dimethyl- 1, 3-dioxolan-4-yl)methyl)-6-nuoro- l H-indol-2-yl)-3-methylbutan-l -ol containing some 3-(5-amino-6-f)uoro-ll l-indol-2-yl)-3-methylbutan- l-ol (220 mg, 0.63 mmol) and I IATU (287 mg, 0.75 mmol) in DMF (3.0 mL) was added triethylamine (0.21 mL, 1.5 mmol). The reaction was stirred at room temperature overnight and then partitioned between ethyl acetate and water. The aqueous layer was extracted with ethyl acetate and the combined organic layers were washed with brine and dried over gSOj. After the removal of solvent, the residue was purified by column chromatography (20-40% ethyl acetate - hexanes) to afford (R)- 1 -(2,2-difluorobenzojd.ll 1 ,3]dioxol-5-y])-N-(l -((2,2-dimethyl- 1 ,3-dioxolan-4-yl)melhyl)-6-fluoro-2-(4-hydroxy-2-methylbutan-2-yl)- 11 l-indol-5-yl)cyclopropanecarboxamide (315 mg, 87 %, contains some l-(2,2-difluorobenzo[d][ l ,3jdioxol-5-y))-N-(6-fluoro-2-(4-hydroxy-2-mcthylbutan-2-yl)-l H-indol-5-yl)cyclopropanecarboxamidc). ESI-MS m ¾ ca]c. 574.2 found 575.7 (M+l)+. Retention lime 2.08 minutes.
[072] Step c: (R)- 1 -(2,2-difluorobenzoi.d][ 1 ,3]dioxol-5-yl)-N-( 1 -(2.3-dihydroxypropyl)-6-fluoro-2-(4-hydroxy-2-melhylbutan-2-yl)-l H-indol-5-yl)cyclopropanecarboxamide
[073] To a solution of (R)-l -(2,2-dif]uorobenzo|d]| l ,3Jdioxol-5-yl)-N-( l - ((2,2-dimethyl- ] ,3-diox()]an-4-y])methy])-6-nuor()-2-(4-hydroxy-2-nnithylbulan-2-y])- 1 H-indol-5-yl)cyclopropanecarboxamide conLaining sonic l -(2,2-diiluorobenzo|d|[ 1 ,3]dioxoi-5-yl)-N-(6-fluoro-2-(4-hydroxy-2-methylbutan-2-yl)- l H-indol-5-yl)cyclopropanecarboxaniide (315 mg, 0.55 mnio!) in methanol (3 mL) and water (0.3 niL) was added p-Ts01I.l l20 (21 mg, 0.1 1 mmol). 'ITie mixture was heated at 80 °C for 30 minutes. The reaction was partitioned between ethyl acetate and water and the aqueous layer was extracted with ethyl acetate twice. The combined organic layers were washed with saturated. N HCOj solution and brine and dried over MgSO,. After the removal of solvent, the residue was purified by column chromatography (20-80% ethyl acetate - hcxancs) to provide (R)- l-(2,2-difluorobenzo[d|[ 1 ,3]dioxol-5-yl)-N-(l -(2,3-dihydroxypropyl)-6-lluoro-2-(4-hydroxy-2-niethylbutan-2-yl)- lH-indol-5-yl)cyclopropanecarboxamide (92 mg, 31 %). ESI- S m/z calc. 534.2, found 535.5 (M+ l)+. Retention lime 1.72 minutes. Ή NMR (400 MHz, DMSO-rf6) δ 8.32 (s, 1 H), 7.53 (d, J = 1.0 Hz, 1 H), 7.43 - 7.31 (m, 4H), 6.17 (s, 1 H), 4.97 - 4.92 (m, 2H), 4.41 (dd, J = 2.4, 15.0 Hz, 111), 4.23 (t, J = 5.0 Hz, I I I), 4.08 (dd, J = 8.6, 15.1 Hz, 1 H), 3.87 (s, 111), 3.48 - 3.44 (m, 1 H), 3.41 - 3.33 (m, 1 H), 3.20 (dd, J = 7.4, 12.7 Hz, 2H), 1.94 - 1.90 (m, 2H), 1.48 - 1.45 (m, 2I-I), 1.42 (s, 311), 1.41 (s, 3H) and 1.15 - 1.12 (m, 2H) ppm. l-(2,2-diHuorobenzotd][l,3]dioxol-5-y-)-N-(2-(4-((S)-2>3-di ydroxypropoxy)- 2-methylbutan-2-yl)-l-((R)-2,3-dihydroxypropyI)-6-fluoro-lH-indoI-5- yl)cyclopropanecarboxamide and (S)-l-(2,2-difluorobenzo[d]tl,3]dioxol-5- yl)-N-(2-(4-(2,3-dihydroxypropoxy)-2-methylbutan-2-yl)-6-fluoro-lH- in d ol-5-y 1 )cyclop ropaneca rboxamide
[074] l-(2,2-difluorobenzo|d]l 1 ,3]dioxol-5-yl)-N-(2-(4-((S)-2,3-dthydroxypropoxy)-2-mcthylbulan-2-yl)-l -((R)-2,3-dihydroxypropyl)-6-nuoro-lH-indol-5-yl)cyclopropanecarboxamide and (S)-l-(2,2-difluorobenzoLdJLl)3 ]dioxol-5-yl)-N-(2-(4-(2,3-dihydroxypropoxy)-2-methylbutan-2-yl)-6-fluoro-l l l-indol-5-yl)cyclopropanecarboxaniide
[075] l-(2,2-difluorobenzo[d]| 1 ,3|dioxol-5-y])-N-(2-(4-((S)-2,3-dihydroxypropoxy)-2-methylbutan-2-yl)- 1 -((R)-2,3-dihydroxypropyl)-6-fluoro- 1 H-indol-5-y cyclopropanecarboxamide and (S)-l-(2,2-difluorobenzo|d] l ,3 ]dioxol-5-yl)-N-(2-(4-(2,3-dihydroxypropoxy)-2-mcthylbutan-2-y])-6-fluoro-lH-indol-5-yl)cyclopropanccarboxaniide were made following a scheme similar as shown above starting from 2-(4-(((R)-2,2-dimelhyl-l,3-dioxolan-4-yl)methoxy)-2-mediylbuto^^ yl)methyl)-6-fluoro-5-nitro- 1 H-indole containing some (R)-2-(4-((2,2-dimethy]- 1 ,3-dioxolan-4-yl)me-hoxy)-2-methylbutan-2-yl)-6-fluoro-5-ni-ro- 1 H-indole). 1 -(2,2-dinuorobenzold || l ,3 |dioxol-5-yl)-N-(2 4-((S)-2,3-dihydroxypropoxy)-2-methyIbutan-2-yl)-l-((R)-2,3-dihydroxypropyl)-6-fluoro-l H-indol-5-yI)eyclopropanecarboxamide, ESI-MS m/z calc. 608.2, found 609.5 (M+ l )+ Retention time 1.67 minutes. Ί-1 NMR (400 MHz, DMSO-d6) δ 8.32 (s, IH), 7.53 (s, I H), 7.43 - 7.31 (m, 4H), 6. 19 (s, I H), 4.95 - 4.93 (m, 2H), 4.51 (d, J = 5.0 Hz, I H), 4.42 - 4.39 (m, 2H), 4.10 - 4.04 (m, I H), 3.86 (s, IH), 3.49 - 3.43 (m, 2H), 3.41 - 3.33 (m, IH), 3.30 - 3.10 (m, 6H), 2.02 - 1.97 (m, 2H), 1 .48 - 1.42 (m, 8H) and 1.13 (dd, J = 4.0, 6.7 Hz, 2H) ppm ; (S)- l-(2,2-difluorobenzold]| l ,3]dioxol-5-yl)-N-(2-(4-(2,3-dihydroxypropoxy)-2-methylbutan-2-yl)-6-lluoro- l H-indol-5-yl)cyclopropanecarboxamide, ESI-MS m/z calc. 534.2, found 535.5 (M+l)+. Retention time 1.81 minutes. Ή NMR (400 MHz, DMSO-rf6) δ 10.9 1 (d, J = 1.5 Hz, H I), 8.30 (s, 111), 7.53 (s, 111), 7.42 - 7.33 (m, 3H), 7.03 (d, J = 10.9 Hz, IH), 6.07 (d, J = 1.6 Hz, II I), 4.56 (d, J = 5.0 Hz, 111), 4.43 (t, J = 5.7 Hz, I H), 3.51 - 3.46 (m, I H), 3.31 - 3.13 (m, 6H), 1 .88 (t, J = 7.3 Hz, 211), 1.48 - 1.45 (m, 211), 1.31 (s, 611) and 1.15 - 1.12 (m, 211) ppm. l-(2,2-difluorobenzo[d][l,3]dioxol-5-yl)-N-(6-fluoro-2-(l-hydroxy-2- methylpropan-2-yl)-lH-indol-5-yl)cyclopropanecarboxaniide
[076] l-(2,2-difluorobenzo|d|[l ,3|dioxol-5-yl)-N-(6-fluoro-2-(l-hydroxy-2-methylpropan-2-yl)-l H-indol-5-yl)cyclopropanecaTboxamide
[077] l -(2,2-dinuorobenzo[d]i l ,3]dioxol-5-yl)-N-(6-nuoro-2-( l -hydroxy-2-methylpropan-2-yl)-l H-indol-5-yl)cyclopropanecarboxamide was made following the scheme shown above starting from a mixture containing (R)-2-(l -((2,2-dimethyM ,3-dioxolan-4-yl)nielhyl)-6-lluoro-5-nilrO-l H-indol-2-yl)-2-methylpropan- l-ol and 3-(6-f1uoro-5-nitro-lH-indol-2-yl)-3-methylbulan-l-ol. ESI-MS m/z calc. 446.2, found 447.5 (M+ l)+. Retention time 1.88 minutes. Ή NMR (400 MHz, CDCI3) 6 8.68 (s, IH), 8.20 (d, J = 7.7 Hz, I H), 7.30 - 7.21 (m, 3H), 7.12 (d, J = 8.2 Hz, I H), 6.94 (d, J = 1 1.2 Hz, I H), 6.18 (s, I H), 3.64 (s, 211), 1.75 (dd, J = 3.8, 6.8 Hz, 2H), 1.34 (s, 611) and 1.14 (dd, J = 3.9, 6.9 Hz, 2H) ppm.
(RJ-l-i^-DifluorobenzoIdJIl^ldioxol-S-y -N-d-il^-dihydroxypropyl)^- fluoro-2-(l-hydroxy-2-methylpropan-2-yl)-lH-indol-5- yOcyclopropanecarboxamide
[078] Step a: (R)-Bcnzyl 2-( l-((2,2-dimcthyl- l ,3-dioxolan-4-y])mcthyl)-6-fluoro-5-nitro- 1 l l-indol-2-yl)-2-methylpropanoate and ((S)-2,2-Di methyl- 1 ,3-dioxolan-4-yl)methyl 2-( l-(((R)-2,2-dimet yl- 1 ,3-dioxolan-4-yl)methyl)-6-f]uoro-5-nitro- 1 H-indol-2-yl)-2-methy 1 propanoate
[079] Cesium carbonate (8.23 g, 25.3 mmol) was added lo a mixture of benzyl 2-(6-fluoro-5-niLro- lH-indol-2-yl)-2-met ylpropanoate (3.0 g, 8.4 mmol) and (S)-(2,2-dimethyl-l ,3-dioxolan-4-yl)methyl 4-methylbenzenesulfonate (7.23 g, 25.3 mmol) in DMF (17 mL). The reaction was stirred at 80 °C for 46 hours under nitrogen atmosphere. The mixture was then partitioned between ethyl acetate and water. The aqueous layer was extracted with ethyl acetate. The combined ethyl acetate layers were washed with brine, dried over S0 , filtered and concentrated. The crude product, a viscous brown oil which contains both of the products shown above, was taken directly to the next step without further purification. (R)-Bcnzyl 2-(l-((2,2-diniclhyl- l ,3-dioxolan-4-yl)methyl)-6-nuoro-5-nilro- lll-indol-2-yl)-2-methylpropanoate, ESI-MS m/z calc. 470.2, found 471.5 (M+lf. Retention time 2.20 minutes. ((S)-2,2-Dimethyl- 1 ,3-dioxolan-4-yl)methyl 2-( 1 -(((R)-2,2-dimethyl- 1 ,3-dioxolan-4-yl)methyi)-6-fluoro-5-nilro- 1 U-indol-2-yl)-2-methylpropanoate, ES1-MS m/z calc. 494.5, found 495.7 (M+ l )+. Retention time 2.01 minutes.
[080] Step b: (R)-2-( l -((2,2-dimethyl- l ,3-dioxolan-4-yl)methyl)-6-fluoro-5-nitro-1 II-indol-2-yl)-2-methylpropan- 1 -ol
[081] To the crude reaction mixture obtained in step (a) was dissolved in THF (42 mL) and cooled in an ice-water bath. LiAll^ ( 16.8 mL of 1 M solution, 16.8 mmol) was added drop- wise. After the addition was complete, the reaction was stirred for an additional 5 minutes. The reaction was quenched by adding water ( I mL), 15% NaOl I solution ( I mL) and then water (3 mL). The mixture was filtered over Celite, and the solids were washed with THF and ethyl acetate. The filtrate was concentrated and purified by column chromatography (30-60% ethyl acetate- hexanes) to obtain the product as a brown oil (2.68g, 87 % over 2 steps). ESI-MS m z cak. 366.4, found 367.3 (M+l )+. Retention time 1.68 minutes. M l N R (400 MHz, DMSO-rf6) δ 8.34 (d, J = 7.6 Hz, 1 H), 7.65 (d, J = 13.4 Hz, 1 H), 6.57 (s, 1H), 4.94 (t, J = 5.4 Hz, 1H), 4.64 - 4.60 (m, 1H), 4.52 - 4.42(m, 211), 4.16 -4. l 4 (m, 1H), 3.76 - 3.74 (m, 1H), 3.63 - 3.53 (m, 2H), 1.42 (s, 311), 1.38 - 1.36 (m, 6H) and 1 .1 (s, 3H) ppm
[082] Step c: (R)-2-(5-aniino- 1-((2,2-dinielhy]- l ,3-dioxolan-4-yl)methyI)-6-fiuoro-1 H-indol-2-yl)-2-methylpropan- 1 -ol
[083] (R)-2-( 1 -((2,2-dimethyl- 1 ,3-dioxolan-4-yl)methyl)-6-fluoro-5-nitro-1 H-indol-2-y1)-2-methylpropan- l -ol (2.5 g, 6.82 mmol) was dissolved ethanol (70 mL) and the reaction was flushed with N2. Then Pd-C (250 mg, 5% wt) was added. The reaction was flushed with nitrogen again and then stirred under H2 (atm). After 2.5 hours only partial conversion to the product was observed by LCMS. The reaction was filtered through Celile and concentrated. The residue was re-subjected to the conditions above. After 2 hours LCMS indicated complete conversion to product. The reaction mixture was filtered through Cclitc. The filtrate was concentrated to yield the product as a black solid (1.82 g, 79 %). ESI-MS m/z calc. 336.2, found 337.5 (M+ l )+. Retention lime 0.86 minutes. Ή NMR (400 MHz, DMSO-i/6) δ 7.17 (d, J = 12.6 Hz, I H), 6.76 (d, J = 9.0 Hz, 1 H), 6.03 (s, 1 H), 4.79 - 4.76 (m, 111), 4.46 (s, 2H), 4.37 - 4.31 (m, 3H),4.06 (dd, J = 6. 1 , 8.3 Hz, 111), 3.70 - 3.67 (m, 1 H), 3.55 - 3.52 (m, 2H), 1.41 (s, 3H), 1.32 (s, 6H) and 1.21 (s, 3H) ppm.
[084] Step d: (R)- 1 -(2,2-difiuorobenzo[d][ 1 ,3 |dioxol-5-yl)-N-(l -((2,2-dimethyl- 1 ,3-dioxolan-4-yl)methy l)-6-fluoro-2-( I -hydroxy-2-methylpropan-2-yl)- 1 H-indol-5-yl)cyclopropanecarboxamide
[085] DMF (3 drops) was added to a stirring mixture of l-(2,2-difluorobenzo|d][l ,3]dioxol-5-yl)cyclopropanecarboxylic acid (1 .87 g, 7.7 mmol) and thionyl chloride ( 1.30 iviL, 17.9 mmol). After 1 hour a clear solution had formed. 'Hie solution was concentrated under vacuum and then toluene (3 mL) was added and the mixture was concentrated again. The toluene step was repeated once more and the residue was placed on high vacuum for 10 minutes. The acid chloride was then dissolved in dichloromethane (10 mL) and added to a mixture of (R)-2-(5-amino- l -((2,2-dimethyl- 1, 3-dioxolan-4-yl)methyl)-6-fluoro-1 H-indol-2-yl)-2-methylpropan- l -ol (1.8 g, 5.4 mmol) and triethyl amine (2.24 mL, 16. 1 mmol) in dichloromethane (45 mL). The reaction was stirred at room temperature for 1 hour. The reaction was washed with I N I ICI solution, saturated NaHCC>3 solution and brine, dried over MgS04 and concentrated to yield the product as a black foamy solid (3g, 100%). liSI- S m/z calc. 560.6, found 561.7 (M+ l )+. Retention time 2.05 minutes. Ή NMR (400 MHz, D SO-rf6) δ 8.31 (s, II I), 7.53 (s, 1H), 7.42 - 7.40 (m, 211), 7.34 - 7.30 (m, 311), 6.24 (s, 1H), 4. 1 - 4.48 (m, I II), 4.39 - 4.34 (m,2H), 4.08 (dd, J = 6.0, 8.3 Hz, 111), 3.69 (l, J = 7.6 Hz, J H), 3.58 - 3.51 (m, 2H), 1.48 - 1.45 (m, 2H), 1.39 (s, 311), 1 .34 - 1.33 (m, 611), 1. 18 (s, 3H) and 1 .14 - 1 .12 (m, 2H) ppm
[086] Step e: (R)- 1 -(2,2-difluorobenzo[d|| 1 ,3 |dioxol-5-yl)-N-( 1 -(2,3-dihydroxypropyl)-6-fluoro-2-(l -hydroxy-2-methylpropan-2-yl)- lH-indol-5-yl)cyelopropanecarboxamide
[087] (R)- l -(2,2-dinuorobenzo[dJ| l ,3 |dioxol-5-yl)-N-( l -((2,2-dimethyl- l ,3-dioxolan-4-yl)methyl)-6-fluoro-2-( l-hydroxy-2-mcthylpropan-2-yl)- lH-indol-5-yl)cyclopropanecarboxamide (3.0 g, 5.4 mmoJ) was dissolved in methanol (52 niL). Water (5.2 niL) was added followed by p-TsOH.H20 (204 mg, 1.1 mmol). The reaction was heated at 80 °C for 45 minutes. The solution was concentrated and then partitioned between ethyl acetate and saturated NaHCCh solution. The ethyl acetate layer was dried over MgS04 and concentrated. The residue was purified by column chronialography (50- 100 % elhyl acetate -hexanes) to yield the product as a cream colored foamy solid. ( 1.3 g, 47 %, ee >98 by SFC). ESI-MS w c calc. 520.5, found 521.7 (M+l)+. Retention time 1.69 minutes. Ή NMR (400 MHz, DMSCM6) δ 8.31 (s, 1 H), 7.53 (s, 111), 7.42 - 7.38 (m, 211), 7.33 - 7.30 (m, 211), 6.22 (s, H I), 5.01 (d, J = 5.2 Hz, 1H), 4.90 (t, J = 5.5 Hz, 1 H), 4.75 (t, J = 5.8 Hz, l H), 4.40 (dd, J = 2.6, 15.1 Hz, 1 H), 4.10 (dd, J = 8.7, 15.1 Yli, I II), 3.90 (s, 1 H), 3.65 - 3.54 (m, 2H), 3.48 - 3.33 Cm, 2H), 1.48 - 1.45 (m, 211), 1.35 (s, 3H), 1.32 (s, 3H) and 1.14 - 1.1 1 (m, 2H) ppm.
(S)-l-(2,2-Difluoroben2o[d][l,3]dioxol-5-yl)-N-(l-(2,3-dihydroxypropyl)-6- fluoro-2-(l-hydroxy-2-methylpropan-2-yl)-lH-indol-5- yl)cyclopropanecarboxamide
[088] Step a: (S)-Benzy 1 2-(l -((2,2-dimethyl- 1 ,3-dioxolan-4-yl)methyl)-6-fluoro-5-nitro-l H-indol-2-yl)-2-tnethylpropanoate and ((R)-2,2-Dimemyl- l ,3-dioxolan-4-yl)methyl 2-(l-(((S)-2,2-dime yl- l ,3-dioxolan-4-yl)me^ niethylpropanoalc
[089] Cesium carbonate (2.74 g, 8.4 mmol) was added to a mixture of benzyl 2-(6-fluoro-5-nitro- l H-indo]-2-y])-2-niethylpropanoaie (1.0 g, 2.8 mmol) and (S)-(2,2-dimethyl- 1 ,3-dioxolan-4-yl)methyl 4-mcthylbenzenesulfonatc (3.21 g, 1 1.2 mmol) in DMF (5.6 mL). I'he reaction was stirred at 80 °C for 64 hours under nitrogen atmosphere. The mixture was then partitioned between ethyl acetate and water. The aqueous layer was extracted with ethyl acetate. The combined ethyl acetate layers were washed with brine, drieii over MgSOj, filtered and concentrated. The crude product, a viscous brown oil which contains both of the products shown above, was taken directly to the next step without further purification. (S)-Benzyl 2-( l -((2,2-dimemyN l,3-dioxolan-4-yl)methyl)-6-fluoro-5-nitro- 1 H-indol-2-yl)-2-methylpropanoate, ESI-MS m/z calc. 470.2, found 471.5 (M+l )+. Retention time 2.22 minutes. ((R)-2,2-Dimethyl- l ,3-dioxolan-4-yl)methyl 2-(l -(((S)-2,2-dimethyl-l ,3-dioxolan-4-yl)niethyl)-6-fluoro-5-nitr - l H-indo!-2-yl)-2-methylpropanoale, ESI-MS m/z calc. 494.5, found 495.5 (M+ 1 )+. Retention time 2.03 minutes.
[090] Step b: (S)-2-( l-((2 DiraeUiyl- l ,3-dioxolan-4-yl)mct y])-6-nuoro-5-niiro-1 H-indol-2-yl)-2-mcthylpropan- 1 -ol [091 ] The mixture of crude reaction mixture of (S)-benzyl 2-( l -((2,2-dimethyl-l,3-dioxolan-4-yl)methyl)-6-fluoro-5-nitro- l H-indol-2-yl)-2-methylpropanoate and ((R)-2,2-dimethyl- l ,3-dioxolan-4-yl)methy] 2-(l -(((S)-2,2-dimethyl-l ,3-dioxolan-4-yl)niethyl)-6-tluoro-5-nitro- l H-indol-2-yl)-2-methylpropanoate was dissolved in THF (15 mL) and cooled in an ice- water bath. UAIH4 (2.8 mL of 1 M solution, 2.8 mmol) was added dropwise. After addition was complete the reaction was stirred for 5 minutes. The reaction was quenched by adding water (0.5 mL), 15% NaOI I solution (0.5 mL) and then water (1.5 mL). The mixture was filtered over Celite, and the solids were washed with THF and ethyl acetate. The filtrate was concentrated and purified by column chromatography (30-60% ethyl acetate- hexanes) to obtain the product as a brown oil (505 mg, 49 % over 2 steps). ESI-MS m z calc. 366.4, found 367.3 (M+ l)+. Retention time 1.68 minutes. Ή NMR (400 MHz, DMSO-i/6) δ 8.34 (d, J = 7.6 Hz, 1H), 7.65(d, J = 13.5 Hz, 1H), 6.57 (s, 1 H), 4.94 (t, J = 5.4 Hz, 1 H), 4.64 - 4.60 (m, 111), 4.52 - 4.42 (m, 2H), 4.14 (dd, J = 6.2, 8.4 Hz, 1 H), 3.74 (dd, J = 7.0, 8.3 Hz, III), 3.63 - 3.53 (m,2H), 1.42 (s, 311), 1.37 (m, 6H) and 1.1 (s, 311) ppm.
[092] Step c: (S)-2-(5-amino- 1 -((2,2-di methyl - 1 ,3-dioxolan-4-yl)methyl)-6-fluoro-I H-indol-2-y l)-2-methy Ipropan- 1 -ol
[093] (S)-2-( l -((2,2-dimethyl-l ,3-dioxolan-4-yl)methyl)-6-fluoro-5-nilro-l H-indol-2-yl)-2-methylpropan- l -ol (500 mg, 1.4 mmol) was dissolved ethanol (15 mL) and the reaction was flushed with N2. Then Pd-C (50 mg, 5% wt) was added. The reaction was flushed with nitrogen again and then stirred under H2 (atni). After 1 hour only partial conversion to the product was observed by LCMS. The reaction was filtered through Celitc and concentrated. J ne residue was resubjected to the conditions above. After 1 hour LCMS indicated complete conversion to product. The reaction mixture was filtered through Celite. The filtrate was concentrated to yield the product as a black solid (420 mg, 1 %). ESI-MS m/z calc. 336.2, found 337.5 (M+ l )+. Retention time 0.90 minutes. Ή NMR (400 MHz, DMSO-d6) δ 7.17 (d, J = 12.6 Hz, 11 1), 6.76 (d, J = 9.0 Hz, I I I), 6.03 (s, I I I), 4.78 (br s, IH), 4.46 (s, 2H), 4.41 - 4.27 (m, 3H), 4.06(dd, J = 6.1 , 8.3 Hz, I H), 3.70 - 3.67 (m, I H), 3.53 (dd, J = 10.7, 17.2 Hz, 2H), 1.40 (s, 3H), 1.32 (s, 6H) and 1.21 (s, 3H) ppm.
[094] Step d: (SH -(2,2-difluorobenzo| d][ 1 ]dioxol-5-yl)-N-( l -((2,2-dimethyl- l ,3-dioxolan-4-yl)methyl)-6-fluoro-2-( 1 -hydroxy-2-mclhylpropan-2-yl)- 1 H-indol-5-y cyclopropanecarboxamide
[095] DMH (3 drops) was added to a stirring mixture of 1 -(2,2-difluorobenzo| d][l,3 ]dioxoL5-yl)cyclopropanecarboxylic acid ( 187 mg, 0.8 mmol) and thionyl chloride (0. 1 3 mL, 1 .8 mmol). After 30 minutes a clear solution had formed. A small amount was mixed piperidine to test that the acid chloride had been formed. The solution was concentrated on the rotovap and then toluene ( 1 mL) was added and the mixture was concentrated again. The toluene step was repeated once more and the residue was placed on high vacuum for 10 minutes. The acid chloride was then dissolved in dichloromethane (2 mL) and added to a mixture of (S)-2-(5-amino- l-((2,2-dimethyl- l ,3-dioxolan-4-yl)melhyl)-6-fluoro- l H-indo!-2-yl)-2-methylpropan- Lol (200 mg, 0.6 mmol) and triethylamine (0.25 mL, 1 .8 mmol) in dichloromethane (4 mL). The reaction was stirred at room temperature for 45 minutes. The reaction was washed with I N HC1 solution, saturated NaHCC^ solution and brine, dried over MgSO
[096] Step e: (S)- 1 -(2,2-difluorobenzo[d]L 1 ,3 |dioxol-5-yl)-N-( 1 -(2,3-dihydroxypropyl)-6-nuoro-2-(l -hydroxy-2-tnelhylpropan-2-yl)- l H-indol-5-y I )cyc lopropanec arboxami de
[097] (S)- 1 -(2,2-difluorobenzoldJL 1 ,3 Jdioxol-5-yl)-N-( l-((2,2-dimethyl- 1 ,3-dioxolan-4-yl)niethyl)-6-nuoro-2-(l-hydroxy-2-melhylpropan-2-yl)- l H-indol-5-yl)cyclopropanecarboxamide (290 g, 0.5 mmol) was dissolved in methanol (5 mL). Water (0.5 mL) was added followed by p-TsOH.H20 (20 mg, 0.1 mmol). The reaction was heated at 80 °C for 45 minutes. The solution was then partitioned between ethyl acetate and saturated NaHCO.i solution. The ethyl acetate layer was dried over MgSC>4 and concentrated. The residue was purified by column chromatography (50- 100 % ethyl acetate - hexanes) to yield the product as a cream colored foamy solid. ( 146 mg, 54 %, ee >97 by SFC). ESI-MS m z calc. 520.5, found 521.5 (M+ l )+. Retention time 1.67 minutes. Ή NMR (400 MHz, D SO-rf6) δ 8.31 (s, 1 H), 7.53 (d, J = 1.1 Hz, 1H), 7.42 - 7.37 (m, 2H), 7.33 - 7.30 (m, 2H), 6.22 (s, 1H), 5.01 (d, J = 5.0 Hz, 1 H), 4.91 (t, J = 5.5 Hz, I H), 4.75 (t, J = 5.8 Hz, 1 H), 4.42 -4.38 (m, 1H), 4.10 (dd, J = 8.8, 15.1 Hz, 1 H), 3.90 (s, 1H), 3.64 - 3.54 (m, 2H), 3.48 - 3.33 (m, 2H), 1.48 - 1.45 (m, 2H), 1.35 (s, 3H), 1.32 (s, 3H) and 1.14 - 1.11 (m, 2H) ppm.
(R)-l-(benzo[d][l,3]dioxol-5-yl)-N-(2-tert-butyl-l-(2,3-dihydroxypropyl)-6- fluoro-lH-indol-5-yl)cyclopropanecarboxamide
[098] (R)- l -(benzo[d][ l ,3]dioxol-5-yl)-N-(2-tert-butyl- l -(2,3-dihydroxypropyl)-6-fluoro- l H-indol-5-yl)cyclopropanecarboxamide was prepared using an experimental procedure similar to example 72 from l -(benzo[d][ l ,3]dioxol-5-yl)cyclopropanccarboxylic acid and 2-tert-bu(yl-6-fluoro-5-nilro- l H-indole.
(S)-l-(benzo[d][l,3]dioxol-5-yl)-N-(2-tert-butyl-l-(2,3-dihydroxypropyl)-6- lluoro-lH-indol-5-yl)cycIopropanecarboxamide
[099] (S)- l -(benzoLdjl 1 ,3Jdioxol-5-yl)-N-(2-tert-buLyl-l -(2,3-dihydrox propyl)-6-fluoro- lH-indol-5-yl)cyclopropanecarboxamide was prepared using an experimental procedure similar to Example 72 from 1 -(benzo[d]| l ,31dioxol-5-yl)cyclopropanecarboxylic acid and 2- (R)-N-(2-tert-butyl-l-(2,3-dihydroxypropyl)-lH-indol-5-yl)-l-(3,4- dihydroxypheny cyclopropanecarboxamide
[0100] (R)-N-(2-terl-butyl- 1 -(2,3-dihydroxypropyl)- 1 H-indol-5-yl)- 1 -(3,4-dihydroxyphenyl)cyclopropanecarboxamide was prepared using an experimental procedure similar to Example 72 from l -(3,4-dihydroxyphenyl)cycIopropanecarboxylic acid and 2-tert-butyl-5-nitro-lll-indolc.
(R)-N-(2-tert-butyl-l-(2,3-dihydroxypropyl)-lH-indol-5-yl)-l-(2,3-dihydro- lH-inden-5-yl)c clopropanecarboxamide [00101S] (R)-N-(2-lert-butyl- l -(2,3-dihydroxypropyl)- lH-indol-5-yl)- l-(2,3-dihydro-lH-inden-5-yl)cyclopropanecarboxamide was prepared using an experimental procedure similar to Example 72 from l -(2,3-dihydro- l H-inden-5-yl)cyclopropanecarboxylic acid and 2-tert-butyl-5-nitro-lH-indolc.
[001016] Λ person skilled in the chemical arts can use the examples and schemes along with known synthetic methodologies to synthesize compounds of the present invention, including the compounds in Table 3, below.
Table 3: Physical data of exemplary compounds.
Assays for Detecting and Measuring AF508-CFTR Correction Properties of Compounds Membrane potential optical methods for assaying AF508-CFTR modulation properties of compounds.
The assay utilizes fluorescent voltage sensing dyes to measure changes in membrane potential using a fluorescent plate reader (e.g., FLIPR 111, Molecular Devices, Inc.) as a readout for increase in functional AF508-CFTR in NIH 3T3 cells. The driving force for the response is the creation of a chloride ion gradient in conjunction with channel activation by a single liquid addition step after the cells have previously been treated with compounds and subsequently loaded with a voltage sensing dye.
Identification of Correction Compounds To identify small molecules that correct the trafficking defect associated with AF508-CFTR; a single-addition HTS assay format was developed. Assay Plates containing cells are incubated for -2-4 hours in tissue culture incubator at 37oC, 5 C02, 90% humidity. Cells are then ready for compound exposure after adhering to the bottom of the assay plates.
The cells were incubated in serum-free medium for 16-24 hrs in tissue culture incubator at 37oC, 5%C02, 90% humidity in the presence or absence (negative control) of test compound. The cells were subsequently rinsed 3X with rebs Ringers solution and loaded with a voltage sensing redistribution dye. To activate AF508-CFTR, 10 μΜ forskolin and the CFTR potentiator, genistein (20 μΜ), were added along with Cl'-free medium to each well. The addition of Cl'-free medium promoted CI" efflux in response to AF508-CFTR activation and the resulting membrane depolarization was optically monitored using voltage sensor dyes.
Identification of Potentiator Compounds To identify potentiators of AF508-CFTR, a double-addition HTS assay format was developed. This HTS assay utilizes fluorescent voltage sensing dyes to measure changes in membrane potential on the FLIPR III as a measurement for increase in gating (conductance) of AF508 CFTR in temperature-corrected AF508 CFTR NIH 3T3 cells. The driving force for the response is a CI" ion gradient in conjunction with channel activation with forskolin in a single liquid addition step using a fluoresecent plate reader such as FLIPR HI after the cells have previously been treated with potentiator compounds (or DMSO vehicle control) and subsequently loaded with a redistribution dye.
Solutions: Bath Solution #1 : (in mM) NaCl 160, C1 4.5, CaCl2 2, MgCl2 1 , HEPES 10, pH 7,4 with NaOH.
Chloride-free bath solution: Chloride salts in Bath Solution #1 are substituted with gluconate salts.
Cell Culture NIH3T3 mouse fibroblasts stably expressing AF508-CFTR are used for optical measurements of membrane potential. The cells are maintained at 37 °C in 5% C02 and 90 % humidity in Dulbecco's modified Eagle's medium supplemented with 2 mM glutamine, 10 % fetal bovine serum, 1 X NEAA, β-ΜΕ, 1 X pen/strep, and 25 mM HEPES in 175 cm2 culture flasks. For all optical assays, the cells were seeded at ~20,000/well in 384-well matrigel-coated plates and cultured for 2 hrs at 37 °C before culturing at 27 °C for 24 hrs. for the potentiator assay. For the correction assays, the cells are cultured at 27 °C or 37 °C with and without compounds for 16 - 24 hours.
Electrophysiological Assays for assaying AF508-CFTR modulation properties of compounds. 1.Ussing Chamber Assay Ussing chamber experiments were performed on polarized airway epithelial cells expressing AF508-CFTR to further characterize the AF508-CFTR modulators identified in the optical assays. Non-CF and CF airway epithelia were isolated from bronchial tissue, cultured as previously described (Galietta. L.J.V., Lantcro, S., Gazzolo, A., Sacco, 0., Romano, L., Rossi, G.A., & Zegarra-Moran, O. (1998) In Vitro Ceil. Dev. Biol. 34, 478-481 ), and plated onto Costar® Snapwell™ filters that were precoated with NIH3T3-conditioned media. After four days the apical media was removed and the cells were grown at an air liquid interface for > 14 days prior to use. This resulted in a monolayer of fully differentiated columnar cells that were ciliated, features that are characteristic of airway epithelia. Non-CF HBE were isolated from non-smokers that did not have any known lung disease. CF-HBE were isolated from patients homozygous for ΔΡ508-CFTR.
HBE grown on Costar® Snapwell'1'" cell culture inserts were mounted in an Ussing chamber (Physiologic Instruments, Inc., San Diego, CA), and the transepithelial resistance and short-circuit current in the presence of a basolateral to apical CI" gradient (ISc) were measured using a voltage-clamp system (Department of Bioengineering, University of Iowa, 1A). Briefly, HBE were examined under voltage-clamp recording conditions (Vhoid = 0 mV) at 37 °C. The basolateral solution contained (in mM) 145 NaCl, 0.83 2HP04, 3.3 H2P04, 1 .2 gCI2, 1 .2 CaCl2, 10 Glucose, 10 HEPES (pH adjusted to 7.35 with NaOH) and the apical solution contained (in mM) 145 NaGluconate, 1.2 MgCl2, 1 .2 CaCl2, 10 glucose, 10 HEPES (pH adjusted to 7.35 with NaOH).
Identification of Correction Compounds Typical protocol utilized a basolateral to apical membrane CI' concentration gradient. To set up this gradient, normal ringer was used on the basolateral membrane, whereas apical NaCl was replaced by equimolar sodium gluconate (titrated to pH 7.4 with NaOH) to give a large CI" concentration gradient across the epithelium. All experiments were performed with intact monolayers. To fully activate AF508-CFTR, forskolin (10 μΜ), PDE inhibitor, IBMX (100 μΜ) and CFTR potentiator, genistein (50 μΜ) were added to the apical side.
As observed in other cell types, incubation at low temperatures of FRT cells and human bronchial epithelial cells isolated from diseased CF patients (CF-HBE)expressing AF508-CFTR increases the functional density of CFTR in the plasma membrane. To determine the activity of correction compounds, the cells were incubated with test compound for 24-48 hours at 37°C and were subsequently washed 3X prior to recording, The cAMP- and genistein-mediated ISc in compound-treated cells was normalized to 37°C controls and expressed as percentage activity of CFTR activity in wt-HBE. Preincubation of the cells with the correction compound significantly increased the cAMP- and genistein-mediated Isc compared to the 37°C controls.
Identification of Potentiator Compounds Typical protocol utilized a basolateral to apical membrane CI" concentration gradient. To set up this gradient, normal ringers was used on the basolateral membrane, whereas apical NaCl was replaced by equimolar sodium gluconate (titrated to pH 7.4 with NaOH) to give a large CI" concentration gradient across the epithelium. Forskolin (10 μΜ) and all test compounds were added to the apical side of the cell culture inserts. The efficacy of the putative AF508-CFTR potentiators was compared to that of the known potentiator, genistein. 2. Patch-clamp Recordings Total Cf current in AF508-NIH3T3 cells was monitored using the perforated-patch recording configuration as previously described (Rae, J., Cooper, K., Gates, P., & Watsky, M. (1991) J. Neurosci. Metiwds 37, 15-26). Voltage-clamp recordings were performed at 22 °C using an Axopatch 200B patch-clamp amplifier (Axon Instruments Inc., Foster City, CA). The pipette solution contained (in mM) 150 N-methyl-D-glucamine (NMDG)-Cl, 2 MgCl2, 2 CaCl2, 10 EGTA, 10 HEPES, and 240 μg ml amphotericin-B (pH adjusted to 7.35 with HC1). The extracellular medium contained (in mM) 150 NMDG-C1, 2 MgCl2, 2 CaCl2, 10 HEPES (pH adjusted to 7.35 with HC1). Pulse generation, data acquisition, and analysis were performed using a PC equipped with a Digidata 1320 A/D interface in conjunction with Ciampex 8 (Axon Instruments Inc.). To activate AF508-CFTR, 10 μΜ forskolin and 20 μΜ genistein were added to the bath and the current-voltage relation was monitored eveiy 30 sec.
Identification of Correction Compounds To determine the activity of correction compounds for increasing the density of functional AF508-CFTR in the plasma membrane, we used the above-described perforated-patch-recording techniques to measure the current density following 24-hr treatment with the correction compounds. To fully activate AF508-CFTR, 10 μΜ forskolin and 20μΜ genistein were added to the cells. Under our recording conditions, the current density following 24-hr incubation at 27°C was higher than that observed following 24-hr incubation at 37 °C. These results are consistent with the known effects of low-temperature incubation on the density of AF508-CFTR in the plasma membrane. To determine the effects of correction compounds on CFTR current density, the cells were incubated with 10 μΜ of the test compound for 24 hours at 37°C and the current density was compared to the 27°C and 37°C controls (% activity). Prior to recording, the cells were washed 3X with extracellular recording medium to remove any remaining test compound. Preincubation with 10 μΜ of correction compounds significantly increased the cAMP- and genistein-dependent current compared to the 37°C controls.
Identification of Potentiator Compounds The ability of AF508-CFTR potentiators to increase the macroscopic AF508-CFTR CI" current (IAFSOS) in NIH3T3 cells stably expressing AF508-CFTR was also investigated using perforated-patch-recording techniques. The potentiators identified from the optical assays evoked a dose-dependent increase in IApsoe with similar potency and efficacy observed in the optical assays. In all cells examined, the reversal potential before and during potentiator application was around -30 mV, which is the calculated Eci (-28 mV).
Cell Culture NIH3T3 mouse fibroblasts stably expressing AF508-CFTR are used for whole-cell recordings. The cells are maintained at 37 °C in 5% C0 and 90 % humidity in Dulbecco's modified Eagle's medium supplemented with 2 mM glutamine, 10 % fetal bovine serum, 1 X NEAA, β-ΜΕ, 1 X pen/strep, and 25 mM HEPES in 175 cm2 culture flasks. For whole-cell recordings, 2,500 - 5,000 cells were seeded on poly-L-lysine-coated glass coverslips and cultured for 24 - 48 hrs at 27 °C before use to test the activity of potentiators; and incubated with or without the correction compound at 37 °C for measuring the activity of correctors, 3.Single-channel recordings Gating activity of wt-CFTR and temperature-corrected AF508-CFTR expressed in NIH3T3 cells was observed using excised inside-out membrane patch recordings as previously described (Dalenians, W., Barbry, P., Champigny, G., Jallat, S., Doll, K., Dreyer, D., Crystal, R.G., Pavirani, A., Lecocq, J-P., Lazdunski, M. (1991 ) Nature 354, 526 - 528) using an Axopatch 200B patch-clamp amplifier (Axon Instruments Inc.). The pipette contained (in mM): 1 0 NMDG, 150 aspartic acid, 5 CaCl2, 2 MgCl2, and 10 HEPES (pH adjusted to 7.35 with Tris base). The bath contained (in mM): 1 50 NMDG -CI, 2 MgCl2, 5 EGTA, 10 TES, and 14 Tris base (pH adjusted to 7.35 with HC1). After excision, both wt- and AF508-CFTR were activated by adding 1 mM Mg-ATP, 75 nM of the catalytic subunit of cAMP-dependent protein kinase (P A; Promega Corp. Madison, WI), and 10 mM NaF to inhibit protein phosphatases, which prevented current rundown. The pipette potential was maintained at 80 mV. Channel activity was analyzed from membrane patches containing≤ 2 active channels. The maximum number of simultaneous openings determined the number of active channels during the course of an experiment. To determine the single-channel current amplitude, the data recorded from 120 sec of AF508-CFTR activity was filtered "off-line" at 100 Hz and then used to construct all-point amplitude histograms that were fitted with multigaussian functions using Bio- Patch Analysis software (Bio-Logic Comp. France). The total microscopic current and open probability (P„) were determined from 120 sec of channel activity. The P0 was determined using the Bio-Patch software or from the relationship P0 = l/i(N), where I = mean current, i = single-channel current amplitude, and N = number of active channels in patch.
Cell Culture NIH3T3 mouse fibroblasts stably expressing AF508-CFTR are used for excised-membrane patch-clamp recordings. The cells are maintained at 37 °C in 5% C02 and 90 % humidity in Dulbecco's modified Eagle's medium supplemented with 2 mM glutamine, 10 % fetal bovine serum, 1 X NEAA, β-ΜΕ, 1 X pen/strep, and 25 mM HEPES in 175 cm1 culture flasks. For single channel recordings, 2,500 - 5,000 cells were seeded on poly-L-lysine-coated glass coverslips and cultured for 24 - 48 hrs at 27 °C before use.
[001017] The compounds of Table I were found to exhibit Correction activity as measured in the assay described above.
[001018] Compounds of the invention are useful as modulators of ATP binding cassette transporters. Using the procedures described above, the activities, i.e., EC50s, of compounds of the present invention have been measured to be from about 3.8 nM to about 13.5 μΜ.
Furthermore, using those methods described above, the efficacies of compounds of the present invention have been measured to be from about 35 % to about 1 10 %.
In Table 4, the following meanings apply: EC50: "+++" means <2 uM; "++" means between 2 uM to 5 uM; 'V means between 5 uM to 25 uM.
% Efficacy: "+" means < 25%; "++" means between 25% and 100%; "+++" means > 100%.
Table 4.
[001019] OTHER EMBODIMENTS
[001020] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims (42)

We claim:
1. A compound of formula II: II or a pharmaceutically acceptable salt thereof, wherein independently for each occurrence: R is H, OH, OCH3 or two R taken together form -CH2CH2CH2-, -OCH20- or -OCF20-; Ri is H or up to two C I -C6 alkyl; R2 is H or F; R3 is H or CN; R4 is H, -CH2CH(0H)CH20H, -CH2CH2N+(CH3)3j or -CH2CH2OH; and Rs is H, OH, -CH2OCH2CH(OH)CH2OH, -CH2OH, or R4 and R5 taken together form a fused pyrrolidine ring.
2. The compound of claim 1 , wherein two R taken together form -OCF20-, Ri is H, and R2 is F.
3. The compound of claim 1 , wherein two R taken together form -OCF20-, R[ is H, R2 is F, and R3 is H.
4. The compound of claim 1 , wherein two R taken together form -OCF20-, Ri is H, R2 is F, R3 is H, and R4 is -CH2CH(OH)CH2OH.
5. The compound of claim 1 , wherein two R taken together form -OCF20-, Ri is H, R2 is F, R3 is H, and R4 is ( )-CH2CH(0H)CH20H.
6. The compound of claim I , having formula Ila: Ila or a pharmaceutically acceptable salt thereof, wherein: 4 is H, -CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is H, OH, -CH2OCH2CH(OH)CH2OH, -CH2OH, or R4 and R5 taken together form a fused pyrrolidine ring.
7. The compound of claim 6, wherein R4 is (i?)-CH2CH(OH)CH2OH, (S)- CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH.
8. The compound of claim 6, wherein R5 is OH, -CH2OCH2CH(OH)CH2OH, or -CH2OH.
9. The compound of claim 6, wherein R4 is (/?)-CH2CH(OH)CH2OH, (5)- CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is OH, - CH2OCH2CH(OH)CH2OH, or -CH2OH.
10. A compound selected from Table 1 .
11. 1 1 . A pharmaceutical composition comprising (i) a compound according to claim 1 ; and (ii) a pharmaceutically acceptable carrier.
12. The composition of claim 1 1 , further comprising an additional agent selected from a mucolytic agent, bronchodialator, an anti-biotic, an anti-infective agent, an antiinflammatory agent, CFTR corrector, CFTR potentiator, or a nutritional agent.
13. A method of increasing the number of functional ABC transporters in a membrane of a cell, comprising the step of contacting said cell with a compound of formula II: II wherein independently for each occurrence: R is H, OH, OCH3 or two R taken together form -CH2CH2CH2-, -OCH20- or -OCF20-; R[ is H or up to two C I -C6 alkyl; R2 is H or F; R3 is H or CN; R4 is H, -CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is H, OH, -CH2OCH2CH(OH)CH2OH, -CH2OH, or 4 and R5 taken together form a fused pyrrolidine ring.
14. The method of claim 13, wherein the ABC transporter is CFTR.
15. 1 5. The method of claim 13, wherein two R taken together form -OCF20-, Ri is H, and R2 is F.
16. The method of claim 13, wherein two R taken together form -OCF20-, Ri is H, R2 is F, and R3 is H.
17. The method of claim 13, wherein two R taken together form -OCF20-, R| is H, R2 is F, R3 is H, and R4 is -CH2CH(OH)CH2OH.
18. The method of claim 13, wherein two R taken together form -OCF20-, | is H, R2 is F, R3 is H, and R4 is (*)-CH2CH(OH)CH2OH.
19. The method of claim 1 3, wherein the compound is represented by formula Ha: or a pharmaceutically acceptable salt thereof, wherein: R4 is H, -CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is H, OH, -CH2OCH2CH(OH)CH2OH, -CH2OH, or R4 and R5 taken together form a fused pyrrolidine ring.
20. The method of claim 1 9, wherein R4 is ( f)-CH2CH(OH)CH2OH, (S)- CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH.
21. . The method of claim 1 9, wherein R5 is OH, -CH2OCH2CH(OH)CH2OH, or -CH2OH.
22. The method of claim 19, wherein R4 is tf)-CH2CH(OH)CH2OH, (S CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is OH, - CH2OCH2CH(OH)CH2OH, or -CH2OH.
23. The method of claim 13, wherein the compound is selected from Table 1 .
24. A method of treating a condition, disease, or disorder in a patient implicated by ABC transporter activity, comprising the step of administering to said patient a compound II or a pharmaceutically acceptable salt thereof, wherein independently for each occurrence: R is H, OH, OCH3 or two R taken together form -CH2CH2CH2-, -0CH2O- or -OCF20-; Ri is H or up to two C1 -C6 alky I; R2 is H or F; R3 is H or CN; RA is H, -CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is H, OH, -CH2OCH2CH(OH)CH2OH, -CH2OH, or R4 and R5 taken together form a fused pyrrolidine ring.
25. The method of claim 24, wherein two R taken together form -OCF20-, Rj is H, and R2 is F.
26. The method of claim 24, wherein two R taken together form -OCF20-} R] is H, R2 is F, and R3 is H.
27. The method of claim 24, wherein two R taken together form -OCF20-, Ri is H, R2 is F, R3 is H, and R4 is -CH2CH(OH)CH2OH.
28. The method of claim 24, wherein two R taken together form -OCF20-, Rt is H, R2 is F, R3 is H, and R4 is (7f)-CH2CH(OH)CH2OH.
29. The method of claim 24, wherein the compound is represented by formula Ha: Ila or a pharmaceutically acceptable salt thereof, wherein: R4 is H, -CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is H, OH, -CH2OCH2CH(OH)CH2OH, -CH2OH, or R4 and R5 taken together form a fused pyrrolidine ring.
30. The method of claim 29, wherein R4 is ( )-CH2CH(OH)CH2OH, (5)- CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH.
31. The method of claim 29, wherein R5 is OH, -CH2OCH2CH(OH)CH2OH, or -CH2OH.
32. The method of claim 29, wherein R4 is ( )-CH2CH(OH)CH2OH, (5)- CH2CH(OH)CH2OH, -CH2CH2"N+(CH3)3, or -CH2CH2OH; and R5 is OH, - CH2OCH2CH(OH)CH2OH, or -CH2OH.
33. The method of claim 24, wherein the compound is selected from Table 1 .
34. The method according to claim 24, wherein said condition, disease, or disorder is selected from cystic fibrosis, hereditary emphysema, hereditary hemochromatosis, coagulation- fibrinolysis deficiencies, such as protein C deficiency, Type 1 hereditary angioedema, lipid processing deficiencies, such as familial hypercholesterolemia, Type 1 chylomicronemia, abetalipoproteinemia, lysosomal storage diseases, such as l-cell disease/pseudo-Hurler, mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II, polyendocrinopathy/hyperinsulemia, diabetes mellitus, laron dwarfism, myleoperoxidase deficiency, primary hypoparathyroidism, melanoma, glycanosis CDG type 1 , hereditary emphysema, congenital hyperthyroidism, osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency, diabetes insipidus (di), neurophyseal di, neprogenic DI, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders such as Huntington, spinocerebullar ataxia type I, spinal and bulbar muscular atrophy, dentatorubal pallidoluysian, and myotonic dystrophy, as well as spongiform encephalopathies, such as hereditary Creutzfeldt-Jakob disease, Fabry disease, Straussler-Scheinker syndrome, COPD, dry-eye disease, and Sjogren's disease.
35. A kit for use in measuring the activity of an ABC transporter or a fragment thereof in a biological sample in vitro or in vivo, comprising: (i) a first composition comprising a compound of formula II: II wherein independently for each occurrence: R is H, OH, OCH3 or two R taken together form -CH2CH2CH2-, -OCH20- or -OCF20-; R) is H or up to two C I -C6 alkyl; R2 is H or F; R3 is H or CN; R4 is H, -CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is H, OH, -CH2OCH2CH(OH)CH2OH, -CH2OH, or R4 and R5 taken together form a fused pyrrolidine ring; and (ii) instructions for: a) contacting the composition with the biological sample; b) measuring activity of said ABC transporter or a fragment thereof.
36. The kit according to claim 35, further comprising instructions for a) contacting an additional composition with the biological sample; b) measuring the activity of said ABC transporter or a fragment thereof in the presence of said additional compound, and c) comparing the activity of the ABC transporter in the presence of the additional compound with the density of the ABC transporter in the presence of said first composition.
37. The kit of claim 35, wherein the kit is used to measure the density of CFTR.
38. The kit of claim 35, wherein the compound is represented by formula Ha: Ha or a pharmaceutically acceptable salt thereof, wherein: R4 is H, -CH2CH(OH)CH2OH, -CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is H, OH, -CH2OCH2CH(OH)CH2OH, -CH2OH, or R4 and R5 taken together form a fused pyrrolidine ring.
39. The kit of claim 38, wherein R4 is (7?)-CH2CH(OH)CH2OH, (S)-CH2CH(OH)CH2OH, - CH2CH2N+(CH3)3i or -CH2CH2OH.
40. The kit of claim 38, wherein R5 is OH, -CH2OCH2CH(OH)CH2OH, or -CH2OH.
41. . The kit of claim 38, wherein R4 is (/?)-CH2CH(OH)CH2OH, (S>CH2CH(OH)CH2OH, - CH2CH2N+(CH3)3, or -CH2CH2OH; and R5 is OH, -CH2OCH2CH(OH)CH2OH, or - CH2 H.
42. The kit of claim 35, wherein the compound is selected from Table 1 . LUZ2ATTO & LUZZATTO By: JSi r¾fc*~>M>>
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