Sepiapterin reductase inhibitors for the treatment of pain

Abstract

Disclosed herein are small molecule heterocyclic inhibitors of sepiapterin reductase (SPR), and prodrugs and pharmaceutically acceptable salts thereof. The Also featured are pharmaceutical compositions of the compounds and uses of these compounds for the treatment or prevention of pain (e.g., inflammatory pain, nociceptive pain, functional pain, and neuropathic pain).

Claims

1. A compound of Formula (II), ##STR00258## or pharmaceutically acceptable salt thereof, wherein X is N or CH; m is 0 or 1; R.sup.1 is (CH.sub.2).sub.nOR.sup.1A, halogen, CN, amino, SO.sub.2R.sup.1A, NHSO.sub.2R.sup.1A, NHC(O)R.sup.1A, or C(O)N(R.sup.1A).sub.2; wherein R.sup.1A is H; n is 0, 1, or 2; R.sup.2 is H or optionally substituted C.sub.1-3 alkyl; R.sup.3 is C(O)R.sup.3A or SO.sub.2R.sup.3A; and R.sup.3A is optionally substituted C.sub.1-6 alkyl.

2. The compound of claim 1, wherein the compound has the following formula, ##STR00259##

3. The compound of claim 1, wherein the compound the following formula, ##STR00260##

4. The compound of claim 1, wherein the compound has the following formula, ##STR00261##

5. The compound of claim 1, wherein X is N, m is 1, and R.sup.2 is H.

6. The compound of claim 5, wherein R.sup.1 is amino.

7. The compound of claim 6, wherein R.sup.3 is C(O)R.sup.3A, and R.sup.3A is C.sub.1-3 alkyl having a C.sub.1-3 alkoxy substituent.

8. The compound of claim 7, wherein the compound is ##STR00262##

9. The compound of claim 1, wherein X is CH, m is 1, and R.sup.2 is H.

10. The compound of claim 9, wherein R.sup.1 is F, OH, CN, CH.sub.2OR.sup.1A, SO.sub.2R.sup.1A, NHSO.sub.2R.sup.1A, or C(O)NH.sub.2; and R.sup.1A is H.

11. The compound of claim 10, wherein R.sup.3 is C(O)R.sup.3A, and R.sup.3A is C.sub.1-3 alkyl comprising a C.sub.1-3 alkoxy substituent.

12. The compound of claim 11, wherein the compound is selected from the group consisting of: ##STR00263##

13. The compound of claim 10, wherein R.sup.3 is SO.sub.2R.sup.3A, and R.sup.3A is optionally substituted C.sub.1-4 alkyl.

14. The compound of claim 13, wherein the compound is selected from the group consisting of: ##STR00264##

15. A method of treating or reducing pain in a mammal, wherein the method comprises the administration of the compound of claim 1, or a pharmaceutically acceptable salt thereof to the mammal in a dosage sufficient to inhibit sepiapterin reductase (SPR).

16. The method of claim 15, wherein the compound is administered in a formulation comprising the compound, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

17. The method of claim 15, wherein the pain is neuropathic, inflammatory, nociceptive, or functional pain.

18. The method of claim 15, wherein the pain is chronic pain.

19. The method of claim 15, wherein the pain is acute pain.

Description

BRIEF DESCRIPTION OF THE DRAWING

(1) FIG. 1 is a schematic of BH4 biosynthesis and control. BH4 is synthesized de novo from guanosine triphosphate (GTP) in three steps mediated by GTP cyclohydrolase (GCH-1), 6-pyruvoyltetrahydriobiopterin synthase (PTPS), and sepiapterin reductase (SPR). BH4 is also generated by a separate recycling pathway that converts quinoid BH4 or BH2 to BH4 via enzymatic reduction.

DETAILED DESCRIPTION

(2) In general, the invention relates to compounds according to Formulas (I) and (II), or a tautomer, prodrug, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, and the use of these compounds and compositions in methods of treatment or to inhibit sepiapterin reductase (SPR).

(3) Compounds of Formula (I) have the following structure

(4) ##STR00016##
or a tautomer, prodrug, or pharmaceutically acceptable salt thereof, where

(5) each of X.sup.1 and X.sup.2 is, independently, N, CH, or C-halogen;

(6) A is a single bond, C(O), or SO.sub.2;

(7) R.sup.1 is (CH.sub.2).sub.nOR.sup.1A, halogen (e.g., F, Cl, Br, or I, preferably Cl), amino (e.g., NH.sub.2), CN, SO.sub.2R.sup.1A, NHSO.sub.2R.sup.1A, NHC(O)R.sup.1A, or C(O)N(R.sup.1A).sub.2;

(8) each R.sup.1A is, independently, H or optionally substituted C.sub.1-6 alkyl;

(9) n is 0, 1, or 2;

(10) R.sup.2 is CH.sub.2OR.sup.2A, optionally substituted C.sub.1-6 alkyl, optionally substituted C.sub.3-9 cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl;

(11) R.sup.2A is H or optionally substituted C.sub.1-6 alkyl;

(12) R.sup.3A and R.sup.3B are both H, or R.sup.3A and R.sup.3B combine to form O;

(13) R.sup.4A and R.sup.4B are both H, or R.sup.4A and R.sup.4B combine to form O;

(14) each of R.sup.5 and R.sup.6 is, independently, H, optionally substituted C.sub.1-6 alkyl, optionally substituted C.sub.3-10 cycloalkyl, optionally substituted alkaryl, or optionally substituted alkheteroaryl; and

(15) where when A is C(O), R.sup.1 is OH, R.sup.2 is CH.sub.2OMe, R.sup.3A, R.sup.3B, R.sup.4A, and R.sup.4B are each H, and R.sup.5 is H, R.sup.6 is not H.

(16) In some embodiments, one and only one of R.sup.3A and R.sup.3B and R.sup.4A and R.sup.4B can combine to form O.

(17) Compounds of Formula (II) have the following structure:

(18) ##STR00017##
or a tautomer, prodrug, or pharmaceutically acceptable salt thereof, where

(19) X is N or CH;

(20) m is 0 or 1;

(21) R.sup.1 is (CH.sub.2).sub.nOR.sup.1A, halogen, CN, amino, SO.sub.2R.sup.1A, NHSO.sub.2R.sup.1A, NHC(O)R.sup.1A, or C(O)N(R.sup.1A).sub.2;

(22) each R.sup.1A is, independently, H or optionally substituted C.sub.1-6 alkyl;

(23) n is 0, 1, or 2;

(24) R.sup.2 is H or optionally substituted C.sub.1-3 alkyl;

(25) R.sup.3 is H, C(O)R.sup.3A, or SO.sub.2R.sup.3A; and

(26) R.sup.3A is optionally substituted C.sub.1-6 alkyl.

(27) Exemplary compounds of the invention, or a tautomer, prodrug, or pharmaceutically acceptable salt thereof, include those shown in Table 1.

(28) TABLE-US-00001 TABLE 1 No. Structure (1) (2) (3) 0 (4) (5) (6) (7) (8) (9) (10) (11) (12) (13) 0 (14) (15) (16) (17) (18) (19) (20) (21) (22) (23) 0 (24) (25) (26) (27) (28) (29) (30) (31) (32) (33) 0 (34) (35) (36) (37) (38) (39)
Synthesis

(29) The compounds described herein, e.g., any of Compounds (1)-(39) or a compound according to Formula (I) or (II), can be prepared according to methods known in the art. Exemplary methods include the following.

(30) Synthesis of Formula (I) Compounds

(31) Compounds of Formula (I) can be synthesized by treating various 2-(1H-indol-3-yl)ethanamine starting materials with electrophiles as shown in Scheme 1.

(32) ##STR00057##
In this scheme, R.sup.2-A-LG is an electrophilic reagent, where R and A are as defined for Formula (I) and LG is a leaving group. Suitable R.sup.2-A-LG reagents include, but are not limited to, alkyl halides, alkyl sulfonates, acyl chlorides, carbonates, acyl anhydrides, and sulfonyl chlorides. Further exemplary reagents are described in the synthetic examples provided herein. In some embodiments, e.g., when R.sup.1 is OH, it may be desirable to selectively deprotect this group (e.g., deacylate any acyl esters that may have formed). Such transformations can be accomplished using methods known in the art, e.g., deprotection under basic conditions, or those described in Greene, Protective Groups In Organic Synthesis, 3.sup.rd Edition (John Wiley & Sons, New York, 1999), which is incorporated herein by reference.

(33) Compounds of Formula (I) can also be prepared from other indole starting materials, as shown in Scheme 2.

(34) ##STR00058##

(35) Using Route A, an indole compound can be elaborated to the corresponding 3-carboxamido intermediate (see, for example, the synthetic protocols for Compounds (10)-(12)). If desired, the carbonyl can be treated under reducing conditions to afford a saturated linking group.

(36) When R.sup.1 is an electron-withdrawing group, Route B can be used. Accordingly, the indole starting material is olefinated to form the corresponding nitroalkene intermediate. Reduction of the nitro group to an amino group followed by treatment with an electrophile R.sup.2-A-LG, as described for Scheme 1, can afford still other compounds of Formula (I). If desired, the indole compound can be N-alkylated using an electrophilic reagent such as R.sup.6-LG, where R.sup.6 is as defined for Formula (I) and LG is a leaving group.

(37) Compounds of Formula (I) can also be prepared by the cyclization of arylhydrazine starting materials as shown in Scheme 3 (see, for example, the synthesis of Compound (17)).

(38) ##STR00059##

(39) Synthesis of Formula (II) Compounds

(40) Compounds of Formula (II) can be prepared by the treatment of phenethylamino starting materials with electrophilic RLG reagents, as shown in Scheme 4.

(41) ##STR00060##
In Scheme 4, R can be any of the R.sup.2 or R.sup.3 groups described for Formula (II), and LG is a leaving group. If a tertiary amine is desired, the number of equivalents of RLG can be adjusted accordingly.

(42) If the required phenethylamino starting material is not commercially available, the required compounds can be prepared from the corresponding phenylcarboxaldehyde via olefination to the corresponding nitroalkene and reduction to form the desired phenethylamino compound (Scheme 5).

(43) ##STR00061##

(44) Compounds of Formula (II) can also be prepared using carboxylic acid starting materials, as shown in Scheme 6.

(45) ##STR00062##
In Scheme 6, Ar represents an aryl group, e.g., phenyl, and Het represents a heteroaryl group, e.g., pyridyl. In this scheme, the carboxylic acid group is transformed to a chloromethyl group. Treatment of this intermediate with cyanide followed by reduction affords the desired amine compound. If required, the amine compound can be treated with an electrophile RLG to afford still other compounds of Formula (II).
Pharmaceutical Compositions

(46) The compounds of the invention (e.g., any of Compounds (1)-(39) or a compound according to Formula (I) or (II)), or tautomers, salts, solvates, or prodrugs thereof, are preferably formulated into pharmaceutical compositions for administration to human subjects in a biologically compatible form suitable for administration in vivo. Accordingly, in another aspect, the present invention provides a pharmaceutical composition that includes a compound of the invention, or a tautomer, salt, solvate, or prodrug thereof, in admixture with a suitable diluent, carrier, or excipient.

(47) The compounds of the invention (e.g., any of Compounds (1)-(39) or a compound according to Formula (I) or (II)) may be used in the form of the free base, in the form of tautomers, salts, solvates, prodrugs, or pharmaceutical compositions. All forms are within the scope of the invention. In accordance with the methods of the invention, the described compounds, or tautomers, salts, solvates, prodrugs, or pharmaceutical compositions thereof, may be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. The compounds of the invention, or tautomers, salts, solvates, prodrugs, or pharmaceutical compositions thereof, may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, or transdermal administration and the pharmaceutical compositions formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.

(48) A compound of the invention (e.g., any of Compounds (1)-(39) or a compound according to Formula (I) or (II)), or a tautomer, salt, solvate, or prodrug thereof, may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsules, or it may be compressed into tablets, or it may be incorporated directly with the food of the diet. For oral therapeutic administration, a compound of the invention, or a tautomer, salt, solvate, or prodrug thereof, may be incorporated with an excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.

(49) A compound of the invention (e.g., any of Compounds (1)-(39) or a compound according to Formula (I) or (II)), or a tautomer, salt, solvate, or prodrug thereof, may also be administered parenterally. Solutions of a compound of the invention can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (200320th edition) and in The United States Pharmacopeia: The National Formulary (USP 32-NF 27), published in 2008.

(50) The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that may be easily administered via syringe.

(51) Pharmaceutical compositions for nasal administration may conveniently be formulated as aerosols, drops, gels, and powders. Aerosol formulations typically include a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which can take the form of a cartridge or refill for use with an atomizing device. Alternatively, the sealed container may be a unitary dispensing device, such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use. Where the dosage form comprises an aerosol dispenser, it will contain a propellant, which can be a compressed gas, such as compressed air or an organic propellant, such as fluorochlorohydrocarbon. The aerosol dosage forms can also take the form of a pump-atomizer.

(52) Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, where the active ingredient is formulated with a carrier, such as sugar, acacia, tragacanth, or gelatin and glycerine. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base, such as cocoa butter.

(53) The compounds of the invention may be administered to an animal, e.g., a human, alone or in combination with pharmaceutically acceptable carriers, as noted above, the proportion of which is determined by the solubility and chemical nature of the compound, chosen route of administration, and standard pharmaceutical practice.

(54) The dosage of the compounds of the invention, and/or compositions comprising a compound of the invention, can vary depending on many factors, such as the pharmacodynamic properties of the compound; the mode of administration; the age, health, and weight of the recipient; the nature and extent of the symptoms; the frequency of the treatment, and the type of concurrent treatment, if any; and the clearance rate of the compound in the animal to be treated.

(55) One of skill in the art can determine the appropriate dosage based on the above factors. The compounds of the invention may be administered initially in a suitable dosage that may be adjusted as required, depending on the clinical response. Generally, dosage levels of between 0.1 g/kg to 100 mg/kg of body weight are administered daily as a single dose or divided into multiple doses. Desirably, the general dosage range is between 250 g/kg to 5.0 mg/kg of body weight per day. Wide variations in the needed dosage are to be expected in view of the differing efficiencies of the various routes of administration. For instance, oral administration generally would be expected to require higher dosage levels than administration by intravenous injection. Variations in these dosage levels can be adjusted using standard empirical routines for optimization, which are well known in the art. In general, the precise therapeutically effective dosage will be determined by the attending physician in consideration of the above identified factors.

(56) Kits

(57) Any of the compounds or pharmaceutical compositions of the invention (e.g., any of Compounds (1)-(39) or a compound according to Formula (I) or (II)) can be used together with a set of instructions, i.e., to form a kit. The kit may include instructions for use of the compounds of the invention in a screening method or as a therapy as described herein. For example, the instructions may provide dosing and therapeutic regimes for use of the compounds of the invention to reduce pain, including any type of pain described herein.

(58) Inhibitors of SPR

(59) The compounds and compositions described herein can be used to inhibit SPR, which catalyzes the final step of the transformation of GTP to BH4. BH4 is an essential co-factor required for normal function of several enzyme and neurotransmitter systems: phenylalanine hydroxylase, tyrosine hydroxylase, tryptophan hydroxylase, and the 3 nitric oxide synthases (NOS) subtypes all rely on BH4 allosteric regulation (Thony et al., Biochem. J. 347:1-16, 2000). BH4 is synthesized from guanosine triphosphate (GTP) in three tightly regulated steps by GCH-1, 6-pyruvoyltetrahydriobiopterin synthase (PTPS), and sepiapterin reductase (SPR) (FIG. 1). The final step in the BH4 synthetic pathway is the conversion of 6-pyruvoyl tetrahydropterin to BH4 by sepiapterin reductase (SPR).

(60) Two of the enzymes involved in de novo BH4 synthesis, GCH-1 and SPR, are up-regulated in preclinical pain models, and reducing the activity of these enzymes leads to preclinical pain relief (Tegeder et al., Nature Medicine 12:1269-1277, 2006). Accordingly, agents that reduce de novo BH4 synthesis (e.g., via direct active site inhibition of SPR) can be used in the prevention or treatment of pain.

(61) Initial studies demonstrated that SPR is weakly inhibited by catecholamines and indoleamines, suggesting a negative feedback mechanism by downstream biogenic amines (Katoh et al., Biochem. Biophys. Res. Commun. 105:75-81, 1982; Smith et al., J. Biol. Chem. 267:5509-5607, 1992). Additional structural information can be obtained by analysis of the SPR protein structure. The crystal structures of human, mouse, and Chlorobium tepidum SPR have been solved in complex with a range of active site ligands including N-acetyl serotonin, NADPH, NADPH+, oxaloacetate and sepiapterin. The first solved structure of mouse SPR reveals a homodimeric structure of 261 amino acids (Auerbach et al., EMBO J. 16:7219-7230, 1997). The liganded protein X-ray crystal structure complexes of SPR reveal an active site formed by a 15 -deep pocket surrounded by the hydrophobic residues Leu105, Leu159, Tyr165, Trp168, Tyr171, Met206 and Cys160. SPR is a homolog of other oxidoreductase drug targets such as the M.tb InhA, the target of anti-tuberculosis drug isoniazid. The inhibition of SPR can be a useful target for developing new methods for the treatment or prevention of pain.

(62) Inhibitors of SPR can be identified according to the methods described herein or known in the art (e.g., Katoh et al., Biochem. Biophys. Res. Commun. 105:75-81, 1982; Smith et al., J. Biol. Chem. 267:5509-5607, 1992).

(63) Although not necessary, if desired, candidate SPR inhibitors can be tested for efficacy in any standard animal model of pain. Various models test the sensitivity of normal animals to intense or noxious stimuli (physiological or nociceptive pain). These tests include responses to thermal, mechanical, or chemical stimuli. Thermal stimuli usually involve the application of hot stimuli (typically varying between 42-55 C.) including, for example: radiant heat to the tail (the tail flick test), radiant heat to the plantar surface of the hindpaw (the Hargreaves test), the hotplate test, and immersion of the hindpaw or tail into hot water. Immersion in cold water, acetone evaporation, or cold plate tests may also be used to test cold pain responsiveness. Tests involving mechanical stimuli typically measure the threshold for eliciting a withdrawal reflex of the hindpaw to graded strength monofilament von Frey hairs or to a sustained pressure stimulus to a paw (e.g., the Ugo Basile analgesiometer). The duration of a response to a standard pinprick may also be measured. When using a chemical stimulus, the response to the application or injection of a chemical irritant (e.g., capsaicin, mustard oil, bradykinin, ATP, formalin, acetic acid) to the skin, muscle joints or internal organs (e.g., bladder or peritoneum) is measured.

(64) In addition, various tests assess pain sensitization by measuring changes in the excitability of the peripheral or central components of the pain neural pathway. In this regard, peripheral sensitization (i.e., changes in the threshold and responsiveness of high threshold nociceptors) can be induced by repeated heat stimuli as well as the application or injection of sensitizing chemicals (e.g., prostaglandins, bradykinin, histamine, serotonin, capsaicin, or mustard oil). Central sensitization (i.e., changes in the excitability of neurons in the central nervous system induced by activity in peripheral pain fibers) can be induced by noxious stimuli (e.g., heat), chemical stimuli (e.g., injection or application of chemical irritants), or electrical activation of sensory fibers.

(65) Various pain tests developed to measure the effect of peripheral inflammation on pain sensitivity can also be used, if desired, to confirm the efficacy of SPR inhibitors (Stein et al., Pharmacol. Biochem. Behav. (1988) 31: 445-451; Woolf et al., Neurosci. (1994) 62: 327-331). Additionally, various tests assess peripheral neuropathic pain using lesions of the peripheral nervous system. One such example is the axotomy pain model (Watson, J. Physiol. (1973) 231:41). Other similar tests include the SNL test which involves the ligation of a spinal segmental nerve (Kim and Chung Pain (1992) 50: 355), the Seltzer model involving partial nerve injury (Seltzer, Pain (1990) 43: 205-18), the spared nerve injury (SNI) model (Decosterd and Woolf, Pain (2000) 87:149), chronic constriction injury (CCI) model (Bennett (1993) Muscle Nerve 16: 1040), tests involving toxic neuropathies such as diabetes (streptozocin model), pyridoxine neuropathy, taxol, vincristine, and other antineoplastic agent-induced neuropathies, tests involving ischaemia to a nerve, peripheral neuritis models (e.g., CFA applied peri-neurally), models of post-herpetic neuralgia using HSV infection, and compression models.

(66) In all of the above tests, outcome measures may be assessed, for example, according to behavior, electrophysiology, neurochemistry, or imaging techniques to detect changes in neural activity. Furthermore, several pain tests that mimic central neuropathic pain involve lesions of the central nervous system including, for example, spinal cord injury (e.g., mechanical, compressive, ischemic, infective, or chemical). In these particular tests, outcome measures are the same as those used for peripheral neuropathic pain.

(67) Therapy and Other Uses

(68) The methods of this invention are useful, for example, for the diagnosis, treatment, reduction, or prevention of various forms of pain.

(69) Pain can take a variety of forms depending on its origin. Pain may be described as being peripheral neuropathic if the initiating injury occurs as a result of a complete or partial transection of a nerve or trauma to a nerve plexus. Alternatively, pain is described as being central neuropathic following a lesion to the central nervous system, such as a spinal cord injury or a cerebrovascular accident. Inflammatory pain is a form of pain that is caused by tissue injury or inflammation (e.g., in postoperative pain or rheumatoid arthritis). Following a peripheral nerve injury, symptoms are typically experienced in a chronic fashion, distal to the site of injury and are characterized by hyperesthesia (enhanced sensitivity to a natural stimulus), hyperalgesia (abnormal sensitivity to a noxious stimulus), allodynia (widespread tenderness associated with hypersensitivity to normally innocuous tactile stimuli), and/or spontaneous burning or shooting lancinating pain. In inflammatory pain, symptoms are apparent, at least initially, at the site of injury or inflamed tissues and typically accompany arthritis-associated pain, musculo-skeletal pain, and postoperative pain. Nociceptive pain is the pain experienced in response to a noxious stimulus, such as a needle prick or during trauma or surgery. Functional pain refers to conditions in which there is no obvious peripheral pathology or lesion to the nervous system. This particular form of pain is generated by abnormal function of the nervous system and conditions characterized by such pain include fibromyalgia, tension-type headache, and irritable bowel syndrome. The different types of pain may coexist or pain may be transformed from inflammatory to neuropathic during the natural course of the disease, as in post-herpetic neuralgia.

(70) The methods of this invention are useful for the diagnosis, treatment, reduction, or prevention of various forms of pain, namely inflammatory pain, nociceptive pain, functional pain, and neuropathic pain, whether acute or chronic. Exemplary conditions that may be associated with pain include, for example, soft tissue, joint, bone inflammation and/or damage (e.g., acute trauma, osteoarthritis, or rheumatoid arthritis), myofascial pain syndromes (fibromyalgia), headaches (including cluster headache, migraine, and tension type headache), myocardial infarction, angina, ischemic cardiovascular disease, post-stroke pain, sickle cell anemia, peripheral vascular occlusive disease, cancer, inflammatory conditions of the skin or joints, diabetic neuropathy, and acute tissue damage from surgery or traumatic injury (e.g., burns, lacerations, or fractures). The present invention is also useful for the treatment, reduction, or prevention of musculo-skeletal pain (after trauma, infections, and exercise), neuropathic pain caused by spinal cord injury, tumors, compression, inflammation, dental pain, episiotomy pain, deep and visceral pain (e.g., heart pain, bladder pain, or pelvic organ pain), muscle pain, eye pain, orofacial pain (e.g., odontalgia, trigeminal neuralgia, glossopharyngeal neuralgia), abdominal pain, gynecological pain (e.g., dysmenorrhea and labor pain), pain associated with nerve and root damage due to trauma, compression, inflammation, toxic chemicals, metabolic disorders, hereditary conditions, infections, vasculitis and autoimmune diseases, central nervous system pain, such as pain due to spinal cord or brain stem damage, cerebrovascular accidents, tumors, infections, demyelinating diseases including multiple sclerosis, low back pain, sciatica, and post-operative pain. Conditions that are amenable to treatment according to the present invention are described in detail, for example, in U.S. Ser. Nos. 10/987,289 and 11/584,449, as well as U.S. Pat. No. 6,593,331, each of which are hereby incorporated by reference.

(71) Combination Therapy

(72) The compounds of the present invention (e.g., any of Compounds (1)(39) or a compound according to Formula (I) or (II)), or a tautomer, salt, solvate, prodrug, or pharmaceutical composition thereof, may be administered either alone or in combination with one or more additional therapeutic agents, such as an analgesic agent used in the treatment of nociception, inflammatory, functional, or neuropathic pain. According to this invention, the second therapeutic agent may or may not produce a therapeutic effect when administered on its own, but results in such an effect (e.g., pain reduction) when administered with the composition of the invention.

(73) Exemplary analgesic agents include, without limitation, nonsteroidal anti-inflammatory agents (NSAIDs) (e.g. rofexocib, celecoxib, valdecoxib, paracoxib, salicylic acid, acetominophen, diclofenac, piroxican indomethacin, ibuprofen, and naproxen), opioid analgesics (e.g., propoxyphene, meperidine, hydromorphone, hydrocodone, oxycodone, morphine, codeine, and tramodol), NMDA antagonist analgesics (e.g., 2-piperdino-1 alkanol derivatives, ketamine, dextormethorphan, eliprodil, or ifenprodil), anesthetic agents (e.g., nitrous oxide, halothane, fluothane), local anesthetics (lidocaine, etidocaine, ropivacaine, chloroprocaine, sarapin, and bupivacaine), benzodiazepines (diazepam, chlordiazepoxide, alprazolam, and lorazepem), capsaicin, tricyclic antidepressants (e.g., amitriptyline, perphanazine, protriptyline, tranylcypromine, imipramine, desimipramine, and clomipramine), skeletal muscle relaxant analgesics (flexeril, carisoprodol, robaxisal, norgesic, and dantrium), migraine therapeutic agents (e.g., elitriptan, sumatriptan, rizatriptan, zolmitriptan, and naratriptan), anticonvulsents (e.g., phenytoin, lamotrigine, pregabalin, carbamazepine, oxcarbazepine, topiramate, valproic acid, and gabapentin), baclofen, clonidine, mexilitene, diphenyl-hydramine, hydroxysine, caffeine, prednisone, methylprednisone, decadron, paroxetine, sertraline, fluoxetine, tramodol, ziconotide, and levodopa.

(74) Further, if desired, the mammal being treated may be administered more than one agent that inhibits the production of BH4 (e.g., those described in U.S. Ser. No. 10/987,289, hereby incorporated by reference). Optionally, the composition of the invention may contain more than one such inhibitor. Alternatively, the mammal may further be administered with specific inhibitors of enzymes that function downstream of BH4, in addition to the composition of the invention.

(75) The following non-limiting examples are illustrative of the present invention.

Examples

(76) Synthesis of Formula (I) Compounds

(77) Synthesis of Compounds (1)-(5)

(78) Compounds (1)-(5) were prepared according to Scheme 7. In this scheme, R can be, for example, any group that is defined for R.sup.2 in Formula (I).

(79) ##STR00063##

(80) General Procedure:

(81) To a cold (0 C.) clear solution of compound A (1.0 mmol) and triethylamine (4.0 mmol) in dichloromethane (25 mL) was added slowly the corresponding acid chloride (2.0 mmol) over 5 minutes. After addition, the reaction mixture was stirred at 0 C. for 5 hours. The reaction mixture was diluted with water (20 mL) and extracted with dichloromethane (220 mL). The combined dichloromethane layers were washed with brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was evaporated to afford crude Intermediate B, which was then used in the next step (Table 2).

(82) TABLE-US-00002 TABLE 2 B1 Compound A (300 mg, 1.41 mmol) was reacted with 2-isopropoxy acetyl chloride (300 mg, 2.82 mmol) in the presence of triethylamine (1.32 mL, 9.40 mmol) and dichloromethane (7.5 mL). Compound B1 was obtained as a pale brown gum (230 mg, crude). Mass (M H): 315.0. B2 Compound A (200 mg, 0.94 mmol) was reacted with cyclopropane carbonyl chloride (0.34 mL, 3.76 mmol) in the presence of triethylamine (mmol) and dichloro- methane (5.0 mL). Com- pound B2 was obtained as a pale brown gum (200 mg, 61%). .sup.1H NMR (CDCl.sub.3): 8.10 (s, 1H), 7.34-7.30 (m, 2H), 7.06 (s, 1H), 6.94-6.91 (m, 1H), 5.67 (bs, 1H), 3.61-3.56 (q, 2H), 2.93 (s, J = 6.63 Hz; 2H), 1.91-1.84 (m, 1H), 1.30-1.16 (m, 4H), 1.05-0.95 (m, 4H), 0.72- 0.67 (m, 2H). Mass (M + H): 313.0. B3 Compound A (300 mg, 1.41 mmol) was reacted with 2-ethoxy acetyl- chloride (206 mg, 1.69 mmol) in the presence of triethylamine (0.4 mL, 2.82 mmol) and dichloro- methane (30.0 mL). Com- pound B3 was obtained as a pale brown gum (200 mg, crude). Mass (M + H): 349.0. B4 Compound A (300 mg, 1.41 mmol) was reacted with 2-isopropoxy acetyl chloride (289 mg, 2.02 mmol) in the presence of triethylamine (0.6 mL, 4.35 mmol) and dichloro- methane (15.0 mL) to give compound B4 as a pale brown gum (230 mg, crude). Mass (M + H): 378.0. B5 Compound A (500 mg, 2.35 mmol) was reacted with morpholine-4- carbonyl chloride (1.08 mL, 9.40 mmol) in the presence of triethyl- amine (1.32 mL, 9.40 mmol) and dichloro- methane (15.0 mL). Compound B5 was ob- tained as a pale brown gum (230 mg, crude).

(83) Preparation of Compounds (1)-(5)

(84) ##STR00069##

(85) General Procedure:

(86) A suspension of Intermediate B (1.0 mmol) and K.sub.2CO.sub.3 (1 mmol) in methanol (20 mL) was stirred at 26 C. for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was diluted with water (30 mL) and extracted with ethyl acetate (220 mL). The combined ethyl acetate layers were washed with water (10 mL), brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the corresponding crude product (Table 3), which was purified by column chromatography (100-200 mesh silica gel) using 3% MeOH in chloroform as eluent.

(87) TABLE-US-00003 TABLE 3 (1) 0 Intermediate B1 (250 mg, 0.761 mmol) was reacted with K.sub.2CO.sub.3 (110 mg, 0.761 mmol) in MeOH (5.0 mL) to give Compound (1) (70 mg, 38.4%) as a pale brown gum. .sup.1H NMR (DMSO-d.sub.6): 10.47 (bs, 1H), 8.5 (s, 1H), 7.82 (bs, 1H), 7.11 (d, J = 8.70 Hz; 1H), 7.01 (s, 1H), 6.82 (s, 1H), 6.59-6.56 (m, 1H), 3.28-3.23 (m, 2H), 2.70 (t, J = 7.46 Hz; 2H), 2.35-2.31 (m, 1H), 1.00 (s, 3H), 0.98 (s, 3H). Mass (M + H): 247.0. IR (cm.sup.1): 3401, 2967, 2928, 1642, 1231, 935, 796. HPLC purity (%): 94.5 (Max plot), 87.98 (254 nm), 96.9 (215 nm). (2) Intermediate B2 (180 mg, 0.51 mmol) was reacted with K.sub.2CO.sub.3 (72 mg, 0.51 mmol) in MeOH (5.0 mL) to give Compound (2) (80 mg, 63%) Pale brown solid. .sup.1H NMR (DMSO-d.sub.6): 10.49 (bs, 1H), 8.59 (s, 1H), 8.16 (t, J = 5.12 Hz; 1H), 7.11 (d, J = 8.78 Hz; 1H), 7.02 (s, 1H), 6.81 (s, 1H), 6.59-6.56 (m, 1H), 3.32-3.27 (m, 2H), 2.71 (t, J = 7.31 Hz; 2H), 1.55-1.49 (m, 1H), 0.69-0.61 (m, 4H). Mass (M + H): 245.0. IR (cm.sup.1): 3420, 2925, 1648, 1456, 1242, 929. HPLC purity (%): 96.32 (Max plot), 96.29 (254 nm), 98.8 (215 nm). (3) Intermediate B3 (250 mg, 0.718 mmol) was reacted with K.sub.2CO.sub.3 (99 mg, 0.718 mmol) in MeOH (5.0 mL) to give Compound (3) (100 mg, 52.9%) Pale brown solid. .sup.1H NMR (DMSO-d.sub.6): 10.49 (bs, 1H), 8.59 (s, 1H), 7.73 (t, J = 5.61 Hz; 1H), 7.11 (d, J = 8.29 Hz; 1H), 7.04 (s, 1H), 6.84 (s, 1H), 6.59-6.57 (m, 1H), 3.81 (s, 2H), 3.47-3.42 (m, 2H), 3.37-3.32 (m, 2H), 2.76 (t, J = 7.56 Hz; 2H), 1.12 (t, J = 6.83 Hz; 3H). Mass (M + H): 263.0. IR (cm.sup.1): 3368, 2921, 1644, 1459, 1374, 671. HPLC purity (%): 97.31 (Max plot), 91.76 (254 nm), 97.63 (215 nm). (4) Intermediate B4 (532 mg, 1.41 mmol) was reacted with K.sub.2CO.sub.3 (195 mg, 1.41 mmol) in MeOH (6.0 mL) to give Compound (4) (150 mg, 38.4%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.91 (bs, 1H), 7.23-7.21 (m, 1H), 7.05- 7.02 (m, 2H), 6.08-6.78 (m, 2H), 5.13 (s, 1H), 3.91 (s, 2H), 3.63-3.53 (m, 3H), 1.09 (s, 3H), 1.08 (s, 3H). Mass (M + H): 277.0. IR (cm.sup.1): 3398, 2972, 1655, 1459, 1213, 935. HPLC purity (%): 98.72 (Max plot), 98.73 (254 nm), 99.23 (215 nm). (5) Intermediate B5 (400 mg, 0.99 mmol) was reacted with K.sub.2CO.sub.3 (137 mg, 0.99 mmol) in MeOH (8.0 mL) to give Com- pound (5) (170 mg, 59%) Pale brown solid. .sup.1H NMR (DMSO-d.sub.6): 10.45 (bs, 1H), 8.56 (s, 1H), 7.10 (d, J = 8.70 Hz; 1H), 7.01 (s, 1H), 6.83 (s, 1H), 6.63-6.56 (m, 2H), 3.53 (t, J = 4.35 Hz; 4H), 3.26-3.25 (m, 6H), 2.73 (t, J = 7.46 Hz; 2H). Mass (M + H): 290.0. IR (cm.sup.1): 3409, 2921, 2853, 1629, 1534, 1263, 1112, 851. HPLC purity (%): 97.27 (Max plot), 98.47 (254 nm), 98.88 (215 nm).
Synthesis of Compounds (6), (7), and (8)

(88) ##STR00075##

(89) Compounds (6)-(8) were each synthesized according to the procedure of Scheme 8. In this procedure, R can be, for example, any group as defined for R.sup.2 in Formula (I).

(90) ##STR00076##

(91) To a cold (0 C.), clear solution of Intermediate C (2.5 g, 11.75 mmol) and triethylamine (8.25 mL, 58.77 mmol) in dichloromethane (80.0 mL) was added slowly actyl chloride (2.67 mL, 37.61 mmol) over 10 minutes. After the addition was complete, the reaction mixture was allowed to warm at room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (20 mL) and washed with water (220 mL) and brine (30 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate D (33 g, 88%) as a pale brown gum. Mass (M+H): 261.0.

(92) Preparation of Intermediate E

(93) ##STR00077##

(94) A suspension of Intermediate D (3.3 g, 12.69 mmol) and K.sub.2CO.sub.3 (1.75 g, 12.69 mmol) in methanol (40.0 mL) was stirred at 26 C. for 2 hours. The reaction mixture was concentrated under reduced pressure; the residue was then diluted with water (50 mL) and extracted with ethyl acetate (250 mL). The combined ethyl acetate layers were washed with water (20 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the corresponding crude Intermediate E (2.3 g, 83.15%), as a pale brown gum. Mass (M+H): 219.0.

(95) Preparation of Intermediate F.

(96) ##STR00078##

(97) To a solution of Intermediate E (2.3 g, 10.55 mmol) in a 30% NaOH solution (1.3 mL) at 26 C., dimethyl sulfate (1.7 mL, 17.93 mmol) was added slowly for 15 minutes. After the addition was complete, the reaction mixture was stirred at ambient temperature for 2 hours. The reaction mixture was acidified with 2N HCl (pH2), diluted with water (20 mL), and extracted with ethyl acetate (250 mL). The combined ethyl acetate layers were washed with water (220 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate F, which was washed with ether (210 ml) and n-pentane (10 mL) and then dried to afford the Intermediate F (1.5 g, 61.4%), as off white solid. .sup.1H NMR (CDCl.sub.3): 8.02 (bs, 1H), 7.26 (s, 1H), 7.04-7.01 (m, 2H), 6.89-6.86 (m, 1H), 5.54 (bs, 1H), 3.86 (s, 3H), 3.60 (q, 2H), 2.94 (t, J=6.73 Hz; 2H), 1.93 (s, 3H). Mass (M+H): 233.0.

(98) Preparation of Intermediate G

(99) ##STR00079##

(100) A solution of Intermediate F (1.4 g, 6.03 mmol) in a 10% NaOH solution (13.5 mL) was stirred at 80 C. for 8 hours. The reaction mixture was basified with 20% NaOH (pH10) and extracted with ethyl acetate (250 mL). The combined ethyl acetate layers were washed with water (30 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product, which was washed with pet ether (210 ml) and n-pentane (10 mL) then dried to afford Intermediate G (1.2 g, crude), as pale brown solid. .sup.1H NMR (CDCl.sub.3): 10.58 (bs, 1H), 7.20 (t, J=8.78 Hz; 1H), 7.07 (s, 1H), 6.98 (s, 1H), 6.71-6.68 (m, 1H), 3.75 (s, 3H), 2.81-2.69 (m, 4H), 1.39 (bs, 2H).

(101) Preparation of Intermediates H1-H3

(102) ##STR00080##

(103) General Procedure:

(104) To a cold (0 C.) solution of Intermediate G (1.0 mmol) and triethylamine (1.5 mmol) in dichloromethane (30 mL), the requisite sulfonyl chloride (1.2 mmol) was added slowly for 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (20 mL), washed with water (210 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate H (Table 4), which was purified by column chromatography (silica gel 100-200 mesh) using 40% ethyl acetate in petroleum ether as eluent.

(105) TABLE-US-00004 TABLE 4 H1 Intermediate G (300 mg, 1.57 mmol) was reacted with methanesulfonyl chloride (271 mg, 2.36 mmol) and triethylamine (0.660 mL, 4.73 mmol) in dichloromethane (20 mL) to give Intermediate H1 (140 mg, crude), as white solid. .sup.1H NMR (CDCl.sub.3): 7.95 (bs, 1H), 7.29 (d, J = 8.78 Hz; 1H), 7.07 (s, 1H), 7.03 (s, 1H), 6.90-6.87 (m, 1H), 4.23 (m, 1H), 3.87 (s, 3H), 3.47 (q, 2H), 3.03 (t, J = 6.54 Hz; 2H), 2.85 (s, 3H). Mass (M + H): 269.1. H2 Intermediate G (230 mg, 1.31 mmol) was reacted with isopropane sulfonyl chloride (225 mg, 1.57 mmol) and triethylamine (0.28 mL, 1.97 mmol) in dichloromethane (20 mL) to give Intermediate H2 (200 mg, crude), as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.97 (bs, 1H), 7.27 (d, J = 8.78 Hz; 1H), 7.07 (s, 1H), 7.03 (s, 1H), 6.89-6.87 (m, 1H), 4.06 (m, 1H), 3.87 (s, 3H), 3.45 (q, 2H), 3.15-3.08 (m, 1H), 3.03 (t, J = 6.63 Hz; 2H), 1.30 (s, 3H), 1.29 (s, 3H). Mass (M + H): 297.0. H3 Intermediate G (400 mg, 2.10 mmol) was reacted with ethylsulfonyl chloride (395 mg, 2.52 mmol) and triethylamine (0.88 mL, 6.31 mmol) in dichloro- methane (20.0 mL) to give Intermediate H3 (400 mg, crude), as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.97 (bs, 1H), 7.27 (d, J = 8.78 Hz; 1H), 7.06 (s, 1H), 7.03 (s, 1H), 6.89-6.87 (m, 1H), 4.18 (m, 1H), 3.87 (s, 3H), 3.43 (q, 2H), 3.03 (t, J = 6.42 Hz; 2H), 2.80 (d, J = 6.63 Hz; 2H), 2.18- 2.11 (m, 1H), 1.02 (s, 3H), 1.01 (s, 3H). Mass (M + H): 311.0.

(106) Preparation of Compounds (6)-(8)

(107) ##STR00084##

(108) General Procedure:

(109) To a cold (40 C.) solution of Intermediate H (1.0 mmol) in dichloromethane (20 mL) was added slowly BBr.sub.3 (4.0 mmol). After the addition was complete, the reaction mixture was allowed to reach 0 C. and then stirred for 2 hours. The reaction mixture was diluted with dichloromethane (20 mL) and washed with water (210 mL). The combined dichloromethane layers were washed with water (10 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product, which was purified by column chromatography (silica gel 100-200 mesh) using 40% ethyl acetate in petroleum ether as eluent (Table 5).

(110) TABLE-US-00005 TABLE 5 (6) Intermediate H1 (130 mg, 0.48 mmol) was reacted with BBr.sub.3 (0.19 mL, 1.95 mmol) in dichloromethane (20.0 mL) to give Com- pound (6) (20 mg, 16%), as white solid. .sup.1H NMR (DMSO-d.sub.6): 10.53 (bs, 1H), 8.62 (s, 1H), 7.13- 7.07 (m, 3H), 6.80 (s, 1H), 6.59-6.57 (m, 1H), 3.19- 3.13 (m, 2H), 2.83-2.76 (m, 5H). Mass (M H): 253.0. IR (cm.sup.1): 3443, 2923, 1635, 1318, 1160, 762. HPLC purity (%): 97.12 (Max plot), 97.84 (254 nm), 97.91 (215 nm). (7) Intermediate H2 (200 mg, 0.67 mmol) was reacted with BBr.sub.3 (0.26 mL, 2.71 mmol) in dichloromethane (20 mL) to give Com- pound (7) (45 mg, 23.5%), as white solid. .sup.1H NMR (DMSO-d.sub.6): 10.51 (bs, 1H), 8.60 (s, 1H), 7.12-7.06 (m, 3H), 6.79 (s, 1H), 6.59-6.56 (m, 1H), 3.19-3.10 (m, 3H), 2.79-2.75 (m, 2H), 1.20 (s, 3H), 1.18 (s, 3H). Mass (M + H): 283.0. IR (cm.sup.1): 3408, 2925, 1460, 1305, 1134, 792. HPLC purity (%): 95.48 (Max plot), 95.68 (215 nm). (8) Intermediate H3 (400 mg, 1.29 mmol) was reacted with BBr.sub.3 (0.5 mL, 5.16 mmol) in dichloromethane (30 mL) to give Compound (8) (85 mg, 22.3%) as brown solid. .sup.1H NMR (DMSO- d.sub.6): 10.52 (bs, 1H), 8.60 (s, 1H), 7.13-7.07 (m, 3H), 6.79 (s, 1H), 6.59-6.57 (m, 1H), 3.18- 3.13 (m, 2H), 2.79-2.75 (m, 4H), 2.05-2.01 (m, 1H), 0.98 (s, 3H), 0.96 (s, 3H). Mass (M + H): 297.0. IR (cm.sup.1): 3424, 2924, 1305, 1138, 793. HPLC purity (%): 98.64 (Max plot), 96.97 (254 nm). 98.37 (215 nm).

(111) Synthesis of Compound (9)

(112) ##STR00088##

(113) Compound (9) was prepared according to the procedure of Scheme 9.

(114) ##STR00089##

(115) To a cold (0 C.) solution of Intermediate C (500 mg, 2.35 mmol) and triethylamine (3.5 mL, 25.51 mmol) in dichloromethane (20.0 mL), was added slowly ethyl sulfonyl chloride (453 mg, 3.52 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (50 mL), washed with water (210 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product, which was purified by column chromatography (silica gel 100-200 mesh) using 30% ethyl acetate in petroleum ether as eluent. Compound (9) was obtained in 15 mg as a pale brown gum. .sup.1H NMR (DMSO-d.sub.6): 10.52 (bs, 1H), 8.61 (s, 1H), 7.16-7.07 (m, 3H), 6.79 (s, 1H), 6.59-6.57 (m, 1H), 3.17-3.12 (m, 3H), 2.97-2.91 (m, 2H), 2.79-2.75 (m, 2H), 1.15 (t, J=7.22 Hz; 3H). Mass (M+H): 268.9. IR (cm.sup.1): 3398, 2925, 1307, 1134, 790. HPLC purity (%): 93.74 (Max plot), 92.54 (215 nm).

(116) Synthesis of Compounds (10)-(12)

(117) ##STR00090##

(118) Compounds (10)-(12) were prepared according to the procedure shown in Scheme 10.

(119) ##STR00091##

(120) To a cold (0 C.) solution of formaldehyde (37% aqueous solution; 4.0 mL, 49.32 mmol) and acetic acid (46.0 g, 762.28 mmol) in dioxane (40.0 mL), was added dimethylamine (40% aqueous solution; 6.4 mL, 58.29 mmol) dropwise for 15 minutes. The reaction was then stirred for an additional 15 minutes. At the same temperature, a solution of Intermediate J (10.0 g, 44.84 mmol) in dioxane (70.0 mL) was added slowly. After the addition was complete, the reaction mixture was stirred at 0 C. for 2 hours. The reaction mixture was diluted with dioxane (100 mL) and then basified (pH10) using an aqueous 10% KOH solution. The obtained solid was collected by filtration, washed with water (350 mL) and dried to afford crude Intermediate K (12.0 g, 95%) as an off white solid, which was used in the next step without further purifications. .sup.1H NMR (DMSO-d.sub.6): 10.72 (bs, 1H), 7.47 (d, J=7.42 Hz; 2H), 7.40-7.29 (m, 3H), 7.23 (d, J=8.78 Hz; 1H), 7.15 (m, 2H), 6.80-6.77 (m, 1H), 5.07 (s, 2H), 3.47 (s, 2H), 2.49 (s, 6H). Mass (M+H): 281.1.

(121) Preparation of Intermediate L

(122) ##STR00092##

(123) To a solution of Intermediate K (14 g, 50.0 mmol) in water (40.0 mL) and ethanol (157.0 mL) at room temperature, sodium cyanide (20.0 g, 408.0 mmol) was added, and the reaction mixture was stirred at 100 C. for 60 hours. The reaction mixture was concentrated; the aqueous residue was then diluted with water (100 mL) and extracted with ethyl acetate (250 mL) to remove the impurities. The aqueous layer was acidified (pH2) using diluted HCl and extracted with dichloromethane (350 mL). The combined organic layers were washed with water (30 mL) and brine (25 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate L (4.5 g, 32%) as pale brown solid. .sup.1H NMR (CDCl.sub.3): 7.98 (bs, 1H), 7.48-7.46 (m, 2H), 7.39-7.28 (m, 4H), 7.18-7.13 (m, 2H), 6.96-6.94 (m, 1H), 5.10 (s, 2H), 3.78 (s, 2H). Mass (M+H): 282.1.

(124) Preparation of Intermediates M1-M3

(125) ##STR00093##

(126) General Procedure:

(127) To a cold (0 C.) solution of Intermediate L (1.0 mmol), EDC.HCl (1.3 mmol), HOBt (1.3 mmol), and triethylamine (1.0 mmol) in dichloromethane (30 mL) was added slowly a solution of corresponding amine (1.1 mmol) in dichloromethane (2 mL) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 16 hours. The reaction mixture was diluted with dichloromethane (25 mL) and washed sequentially with water (10 mL), 10% NaHCO.sub.3 solution (10 mL), water (10 mL), and brine solution (20 mL). The organic layer was then dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate M (Table 6), which was purified by column chromatography (silica gel 100-200 mesh) using 1% MeOH in chloroform as eluent.

(128) TABLE-US-00006 TABLE 6 M1 Intermediate L (125 mg, 0.445 mmol) was reacted with methoxy ethylamine (37 mg, 0.49 mmol), EDCHCl (111 mg, 0.577 mmol), HOBt (78 mg, 0.577 mmol), and Et.sub.3N (0.063 mL, 0.45 mmol) to give Intermediate M1 (110 mg, 73%), off white solid. .sup.1H NMR (CDCl.sub.3): 8.09 (bs, 1H), 7.48-7.46 (m, 2H), 7.39-7.28 (m, 4H), 7.12-7.07 (m, 2H), 6.98- 6.96 (m, 1H), 6.04 (b, 1H), 5.09 (s, 2H), 3.70 (s, 2H), 3.37-3.32 (m, 4H), 3.18 (s, 3H). Mass (M + H): 339.2. M2 Intermediate L ((500 mg, 1.779 mmol) was reacted with 2-aminopyridine (185 mg, 1.96 mmol), EDCHCl (442 mg, 2.31 mmol), HOBt (312 mg, 2.31 mmol), and Et.sub.3N (0.25 mL, 1.78 mmol) to give Inter- mediate M2 (180 mg, 29%), off white solid. .sup.1H NMR (CDCl.sub.3): 8.26 (d, J = 8.59 Hz; 1H), 8.15 (m, 2H), 8.00 (bs, 1H), 7.70-7.66 (m, 1H), 7.45-7.43 (m, 2H), 7.36- 7.25 (m, 4H), 7.22-7.20 (s, 1H), 7.11 (s, 1H), 7.00-6.96 (m, 2H), 5.07 (s, 2H), 3.88 (s, 2H). Mass (M + H): 358.2. M3 Intermediate L ((500 mg, 1.779 mmol) was reacted with 3-aminopyridine (185 mg, 1.96 mmol), EDCHCl (442 mg, 2.31 mmol), HOBt (312 mg, 2.31 mmol), and Et.sub.3N (0.25 mL, 1.78 mmol) to give Intermediate M3 (180 mg, 35%), off white solid. .sup.1H NMR (CDCl.sub.3): 8.31-8.25 (m, 1H), 8.12-8.03 (m, 2H), 7.45-7.43 (m, 2H), 7.36-7.32 (m, 2H), 7.28- 7.20 (m, 4H), 7.09 (m, 1H), 7.03-7.00 (m, 1H), 5.08 (s, 2H), 3.88 (s, 2H). Mass (M + H): 358.1.

(129) Preparation of Intermediates N2 and N3

(130) ##STR00097##

(131) General Procedure:

(132) To a cooled solution of intermediate M (1.0 mmol) in THF (30 mL) at 0 C., a solution of BH.sub.3DMS (15.0 mmol) was added at 0 C. After the addition was complete, the reaction mixture was stirred at 70 C. for 2 hours. The reaction mixture was cooled to 0 C., quenched with a mixture of methanol (2.0 mL) and 2N HCl (5.0 mL). After refluxing for 1 hour, the solvent was evaporated and the aqueous residue was basified (pH10) using 2N NaOH solution. The aqueous layer was extracted with ethyl acetate (250 mL), washed with water (215 mL) and brine (15 mL), dried over dried over anhydrous Na.sub.2SO.sub.4, and evaporated to yield the crude product, which was purified by column chromatography (silica gel 100-200 mesh) using 80% ethyl acetate in pet ether as eluent to afford Intermediate N (Table 7).

(133) TABLE-US-00007 TABLE 7 N2 Intermediate M2 (150 mg, 0.42 mmol) was reacted with BH.sub.3DMS (0.6 mL, 6.30 mmol) in THF (20.0 mL) to give Intermediate N2 (75 mg, 52%), off white solid. .sup.1H NMR (CDCl.sub.3): 8.25 (bs, 1H), 8.09-8.08 (m, 1H), 7.47- 7.23 (m, 7H), 7.14 (s, 1H), 6.98-6.92 (m, 2H), 6.57- 6.54 (m, 1H), 6.35 (d, J = 8.29 Hz; 1H), 5.07 (s, 2H), 4.61 (bs, 1H), 3.60-3.56 (q, 2H), 3.03 (t, J = 6.83 Hz, 2H). Mass (M + H): 344.2. N3 Intermediate M3 (300 mg, 0.84 mmol) was reacted with BH.sub.3DMS (1.15 mL, 12.63 mmol) in THF (20.0 mL) to give intermediate N3 (120 mg, 42%), off white solid. .sup.1H NMR (CDCl.sub.3): 8.04-7.94 (m, 3H), 7.47-7.25 (m, 6H), 7.09-7.03 (m, 3H), 6.96 (d, J = 8.78 Hz; 1H), 6.87-6.84 (m, 1H), 5.08 (s, 2H), 3.78 (bs, 1H), 3.44 (bs, 2H), 3.05 (t, J = 6.54 Hz, 2H). Mass (M + H): 344.2.

(134) Preparation of Compounds (11) and (12)

(135) ##STR00100##

(136) General Procedure:

(137) To a cold (70 C.) solution of Intermediate O (1.0 mmol) in dichloromethane (30 mL), BCl.sub.3 (0.1 M in DCM) (1.4 mmol) was added slowly. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 1 hour. The reaction mixture was diluted with dichloromethane (20 mL), washed with water (220 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product (Table 8), which was purified by PREP-TLC using 6% MeOH in chloroform as eluent.

(138) TABLE-US-00008 TABLE 8 (11) 01 Intermediate N2 (75 mg, 0.218 mmol) was reacted with BCl.sub.3 (3.05 mL, 0.305 mmol) in dichloromethane (10 mL) to give Com- pound (11) (18 mg, 30%) as an off white solid. .sup.1H NMR (DMSO-d.sub.6): 10.48 (bs, 1H), 8.58 (s, 1H), 7.98 (m, 1H), 7.34 (m, 1H), 7.12 (d, J = 8.70 Hz; 1H), 7.06 (s, 1H), 6.86 (s, 1H), 6.60- 6.43 (m, 4H), 3.49-3.44 (m, 2H), 2.85-2.81 (m, 2H). Mass (M + H): 254.1. IR (cm.sup.1): 3406, 2920, 2854, 1606, 1210, 1095, 771. HPLC purity (%): 96.37 (Max plot), 96.26 (254 nm), 96.18 (215 nm). (12) 02 Intermediate N3 (120 mg, 0.348 mmol) was reacted with BCl.sub.3 (5.0 mL, 0.5 mmol) in dichloromethane (15.0 mL) to give Com- pound (12) (25 mg, 28%) as an off white solid. .sup.1H NMR (DMSO-d.sub.6): 10.52 (bs, 1H), 8.60 (s, 1H), 7.98 (s, 1H), 7.75 (m, 1H), 7.13-7.05 (m, 3H), 6.91 (d, J = 8.29 Hz; 1H), 6.82 (s, 1H), 6.60-6.57 (m, 1H), 5.91 (s, 1H), 3.30-3.25 (m, 2H), 2.87-2.83 (m, 2H). Mass (M + H): 254.1. IR (cm.sup.1): 3398, 2919, 2851, 1586, 1467, 791. HPLC purity (%): 95.72 (Max plot), 94.43 (254 nm), 96.01 (215 nm).

(139) Preparation of Compound (10)

(140) ##STR00103##

(141) A suspension of Intermediate M1 (100 mg, 0.29 mmol) and 10% Pd/C (30 mg, dry) in MeOH (10.0 mL) was hydrogenated (50 psi H.sub.2 pressure) at room temperature for 5 hours. The reaction mixture was filtered, and the cake was washed with methanol (35 mL). The combined filtrates were concentrated under reduced pressure to give crude Compound (10), which was purified by PREP-TLC using 5% MeOH in chloroform as eluent to afford the product (30 mg, 41%) as an off white solid. .sup.1H NMR (DMSO-d.sub.6): 10.54 (bs, 1H), 8.57 (s, 1H); 7.85 (s, 1H), 7.11 (d, J=8.78 Hz; 1H), 7.06 (s, 1H), 6.83 (s, 1H), 6.58 (dd, 1H), 3.40 (s, 2H), 3.34-3.30 (m, 2H), 3.22-3.17 (m, 5H). Mass (M+H): 249.1. IR (cm.sup.1): 3378, 3323, 2934, 1641, 1228, 1019, 669. HPLC purity (%): 99.73 (Max plot), 99.71 (215 nm).

(142) Preparation of Compounds (13)-(15)

(143) ##STR00104##

(144) Compounds (13)-(15) can be synthesized according to the procedure shown in Scheme 10 by using one of the following amines in the preparation of Intermediate M (Table 9).

(145) TABLE-US-00009 TABLE 9 Compound Amine reagent (13) 05 (14) 06 (15) 07
Synthesis of Compound (16)

(146) ##STR00108##

(147) Compound (16) was prepared according to the procedure in Scheme 11.

(148) ##STR00109##

(149) To a cold (0 C.) solution of Intermediate O (250 mg, 1.16 mmol) and triethylamine (0.32 mL, 2.32 mmol) in dichloromethane (5.0 mL) was added slowly methoxyacetyl chloride (0.12 mL, 1.39 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (20 mL), washed with water (220 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. The material was then washed with petroleum ether (24 mL), n-pentane (3 mL), and dried to afford Compound (16) (20 mg, 68.2%) as pale yellow solid. .sup.1H NMR (CDCl.sub.3): 8.15 (bs, 1H), 7.29-7.22 (m, 2H), 7.09 (s, 1H), 6.97-6.93 (m, 1H), 6.68 (bs, 1H), 3.87 (s, 2H), 3.64-3.59 (m, 2H), 3.33 (s, 3H), 2.95 (t, J=6.84, Hz; 2H). Mass (M+H): 251.0. HPLC purity (%): 98.87 (Max plot), 98.00 (254 nm), 98.88 (215 nm).

(150) Synthesis of Compound (17)

(151) ##STR00110##

(152) Compound (17) was prepared according the procedure described in Scheme 12.

(153) ##STR00111##

(154) Preparation of Intermediate S

(155) ##STR00112##

(156) A solution of Intermediate P (1.0 g, 5.53 mmol) in concentrated HCl (0.2 mL) and water (11.6 mL) was stirred at room temperature for 1 hour. A solution of Intermediate Q (1.1 g, 4.97 mmol) in water (2.4 mL) and MeOH (12.8 mL) was added to the above mixture, which was then stirred at room temperature for 1 hour. The reaction mixture was cooled to 0 C. and the solid was filtered off, washed with 9:1 aqueous methanol (5.0 mL) and water (10.0 mL), and dried. To the solution of this compound in water (7.2 mL) and MeOH (29.0 mL) was then added Na.sub.2HPO.sub.4 (0.5 g, 3.54 mmol) and concentrated HCl (0.1 mL). The reaction mixture was then stirred at reflux for 20 hours. The reaction mixture was concentrated; the aqueous residue was then diluted with water (20 mL), saturated with Na.sub.2CO.sub.3, and extracted with dichloromethane (325 mL). The combined dichloromethane layers were dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate S, which was purified by column chromatography (silica gel 100-200 mesh) using 2% (MeOH/NH.sub.3) in chloroform as eluent to afford the product (80 mg, 6%) as brown solid. .sup.1H NMR (DMSO-d.sub.6): 11.41 (bs, 1H), 8.12 (s, 1H), 7.59-7.53 (m, 2H), 7.38 (s, 1H), 3.15 (s, 3H), 2.83 (s, 4H). Mass (M+H): 239.0.

(157) Preparation of Compound (17)

(158) ##STR00113##

(159) To a cold (0 C.) solution of Intermediate S (150 mg, 0.63 mmol) and triethylamine (0.13 mL, 0.94 mmol) in dichloromethane (10.0 mL) was added slowly a solution of methoxyacetyl chloride (0.06 mL, 0.69 mmol) in dichloromethane (2.0 mL) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (20 mL), washed with water (220 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was then purified by column chromatography (silica gel 100-200 mesh) using 2% MeOH in chloroform as eluent to afford Compound (17) (80 mg, 41%) as off white solid. .sup.1H NMR (CDCl.sub.3): 8.40 (bs, 1H), 8.25 (s, 1H), 7.75 (d, J=8.39 Hz; 1H), 7.50 (d, J=8.78 Hz; 1H), 7.24 (s, 1H), 6.66 (bs, 1H), 3.89 (s, 2H), 3.68-3.63 (m, 2H), 3.36 (s, 3H), 3.09 (s, 3H), 3.06-3.03 (t, J=7.03 Hz; 2H). Mass (MH): 309.0. IR (cm.sup.1): 3344, 2925, 1656, 1289, 1147, 750. HPLC purity (%): 98.90 (Max plot), 94.95 (254 nm), 97.96 (215 nm).

(160) Synthesis of Compound (18)

(161) ##STR00114##

(162) Compound (18) was synthesized according to the procedure shown in Scheme 13.

(163) ##STR00115##

(164) Preparation of Intermediate T

(165) ##STR00116##

(166) To a solution of Intermediate T (2.5 g, 15.41 mmol) in ether (50.0 mL) was added a solution of oxalyl chloride (5.4 mL, 61.67 mmol) in ether (15.0 mL) at room temperature. The resulting reaction was stirred at 40 C. for 16 hours. The reaction mixture was filtered, and the resulting solid was washed with ether (220 mL). The solid was then added to a saturated solution of NH.sub.3 in dioxane (100.0 mL) and stirred for 4 hours. The reaction mixture was basified with Na.sub.2CO.sub.3, and concentrated to yield the crude product, which was purified by column chromatography (silica gel 100-200 mesh) using 70% ethyl acetate in petroleum ether as eluent to afford Intermediate U (1.2 g, 31%) pale yellow solid. .sup.1H NMR (DMSO-d.sub.6): 12.75 (bs, 1H), 9.08 (s, 1H), 8.93 (s, 1H), 8.19-8.15 (m, 2H), 7.86 (s, 1H), 7.74 (d, J=8.78 Hz; 1H). Mass (MH): 232.0.

(167) Preparation of Intermediate V

(168) ##STR00117##

(169) To a suspension of lithium aluminum hydride (3.26 g, 85.0 mmol) in THF (150.0 mL) at room temperature was added a solution of Intermediate U (1.0 g, 4.29 mmol) was added. The resulting mixture was stirred at 70 C. for 48 hours. The reaction mixture was then cooled to 0 C., quenched with ice cold water (5.0 mL), and filtered. The cake was washed with ethyl acetate (3100 mL), and the combined filtrates were extracted. The separated organic layer was washed with water (25 mL) and brine (15 mL), dried over dried over anhydrous Na.sub.2SO.sub.4, and evaporated to yield crude Intermediate V, which was purified by column chromatography (silica gel 100-200 mesh) using (5:94:1 to 20:79:1) MeOH:chloroform:aq NH.sub.3 as the eluent to afford the product (400 mg, 53.2%) as a brown gum. .sup.1H NMR (DMSO-d.sub.6): 10.30 (s, 11H), 7.22-7.00 (m, 2H), 6.93 (s, 1H), 6.47-6.44 (dd, 1H), 4.39 (bs, 2H), 2.81 (t, J=7.07 Hz; 2H), 2.66 (t, J=7.31 Hz; 2H). Mass (M+H): 176.1.

(170) Preparation of Intermediate W

(171) ##STR00118##

(172) To a cold (0 C.) solution of Intermediate V (500 mg, 2.86 mmol) in acetic acid (15.0 mL) was added slowly a solution of NaNO.sub.2 (217 mg, 3.14 mmol) in cold water (1.6 mL) over 5 minutes. After stirring for 5 minutes, a solution of NaN.sub.3 (204 mg, 3.14 mmol) in cold water (1.6 mL) was added slowly. After the addition was complete, the reaction mixture was stirred at 0 C. for 2 hours. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by column chromatography (silica gel 100-200 mesh) using (5:94:1 to 20:79:1) MeOH:chloroform:(aqueous NH.sub.3) as the eluent to afford Intermediate W (220 mg, 38%) as a brown solid. .sup.1H NMR (DMSO-d.sub.6): 10.97 (bs, 1H), 7.42-7.32 (m, 1H), 7.21 (m, 2H), 6.82-6.80 (m, 1H), 2.79-2.74 (m, 4H). Mass (M+H): 202.1. IR (cm.sup.1): 3433, 2918, 2106, 920, 792.

(173) Preparation of Intermediate X

(174) ##STR00119##

(175) To a cold (0 C.) solution of Intermediate W (200 mg, 0.995 mmol) and triethylamine (20.14 mL, 0.995 mmol) in dichloromethane (20 mL) was added slowly a solution of methoxyacetyl chloride (100 mg, 0.895 mmol) in dichloromethane (5.0 mL) over 30 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 1 hour. The reaction mixture was diluted with dichloromethane (30 mL), washed with water (215 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product, which was purified by column chromatography (silica gel 100-200 mesh) using 20% MeOH in chloroform as the eluent to afford Intermediate X (200 mg, 74%) as an off white solid. .sup.1H NMR (CDCl.sub.3): 8.08 (bs, 1H), 7.34 (d, J=8.59 Hz; 1H), 7.24 (s, 1H), 7.09 (s, 1H), 6.90 (dd, 1H), 6.64 (b, 1H), 3.88 (s, 2H), 3.65-3.60 (q, 2H), 3.35 (s, 3H), 2.96 (t, J=6.93 Hz; 2H). Mass (M+H): 274.1.

(176) Preparation of Intermediate Y

(177) ##STR00120##

(178) A suspension of Intermediate X (200 mg, 0.73 mmol) and 10% Pd/C (30 mg, dry) in MeOH (20.0 mL) was hydrogenated (30 psi H.sub.2 pressure) at room temperature for 1 hour. The reaction mixture was filtered through Celite, and the cake was washed with methanol (35 mL). The combined filtrates were concentrated under reduced pressure to give crude Intermediate Y (180 mg, crude) brown gum. .sup.1H NMR (DMSO-d.sub.6): 10.31 (bs, 1H), 7.80 (t, J=5.39 Hz; 1H), 7.01 (d, J=8.29 Hz; 1H), 6.95 (s, 1H), 6.66 (s, 1H), 6.46 (dd, 1H), 4.42 (m, 2H), 3.78 (s, 2H), 3.41-3.28 (m, 5H), 2.72 (t, J=7.46 Hz; 2H). Mass (M+H): 248.1.

(179) Preparation of Compound (18)

(180) ##STR00121##

(181) To a cold (0 C.) solution of Intermediate Y (90 mg, 0.364 mmol) and triethylamine (0.06 mL, 0.40 mmol) in dichloromethane (10.0 mL) was added slowly a solution of methanesulfonyl chloride (0.03 mL, 0.327 mmol) in dichloromethane (2.0 mL) over 15 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 6 hours. The reaction mixture was diluted with dichloromethane (30 mL), washed with water (210 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product, which was purified by PREP-TLC using 5% MeOH in chloroform as the eluent to afford Compound (18) (35 mg, 30%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 8.12 (bs, 1H), 7.52 (s, 1H), 7.35 (d, J=8.35 Hz; 1H), 7.14-7.11 (m, 2H), 6.66 (bs, 1H), 6.33 (bs, 1H), 3.88 (s, 2H), 3.63-3.60 (q, 2H), 336 (s, 3H), 3.00-2.96 (m, 5H). Mass (M+H): 326.1. IR (cm.sup.1): 3387, 3275, 2930, 1658, 1321, 1148, 975. HPLC purity (%): 97.97 (Max plot), 97.20 (215 nm).

(182) Synthesis of Compounds (19) and (20)

(183) ##STR00122##

(184) Compounds (19) and (20) were synthesized according to Scheme 14.

(185) ##STR00123##

(186) Preparation of Intermediate AA

(187) ##STR00124##

(188) POCl.sub.3 (3.6 mL, 38.68 mmol) was added to DMF (16.5 mL) dropwise at 0 C.-10 C. The resulting mixture was stirred for 30 minutes, cooled to 0 C., and a solution of Intermediate Z (5.0 g, 35.17 mmol) in DMF (10.0 mL) was added over 15 minutes. After the addition was complete, the reaction mixture was stirred at ambient temperature for 2 hours. The reaction mixture was quenched with ice (25 g), poured into water (50 mL), and NaOH (1.5 g) was added. The mixture was filtered, and the yellow colored filtrate was diluted with water (100 mL) and left to stand at room temperature for 20 hours. The solid was then filtered and dried to afford Intermediate AA (1.5 g, 62%) as a yellow solid. .sup.1H NMR (DMSO-d.sub.6): 12.59 (bs, 1H), 10.00 (s, 1H), 8.52 (s, 1H), 8.51 (s, 1H), 7.71 (s, J=8.29 Hz; 1H), 7.65 (d, J=8.70 Hz; 1H). Mass (MH): 169.1.

(189) Preparation of Intermediate AB

(190) ##STR00125##

(191) A suspension of Intermediate AA (4.2 g, 24.7 mmol) and ammonium acetate (4.18 g, 54.34 mmol) in nitromethane (263.0 mL) was stirred at 90 C. for 2 hours. The reaction mixture was concentrated under reduced pressure. The crude product was washed with 25% ethyl acetate in petroleum ether (220 mL) and dried to afford Intermediate AB (3.5 g, 68%) as yellow solid. .sup.1H NMR (DMSO-d.sub.6): 10.84 (bs, 1H), 8.67 (s, 1H), 8.43-8.40 (m, 2H), 8.23 (d, J=13.66 Hz; 1H), 7.68 (d, J=8.29 Hz; 1H), 7.61 (d, J=8.78 Hz; 1H). Mass (MH): 212.0.

(192) Preparation of Intermediate AC

(193) ##STR00126##

(194) To a cold (0 C.) solution of Intermediate AB (3.5 g, 16.4 mmol) in methanol and DMF (1:1; 35 mL) was added portionwise NaBH.sub.4 (7.08 g, 18.73 mmol) over 15 minutes. After the addition was complete, the reaction mixture was allowed to reach 10 C. and stirred for 2 hours.

(195) The reaction mixture was quenched with water (10 mL) and concentrated under reduced pressure. The resulting aqueous residue was diluted with water (25 mL) and extracted with ethyl acetate (250 mL). The combined ethyl acetate layers were washed with water (30 mL) and brine solution (25 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AC, which was purified by column chromatography (silica gel 100-200 mesh) using 20% ethyl acetate in petroleum ether as the eluent to afford the product (2.5 g, 71.4%) as pale yellow solid. .sup.1H NMR (CDCl.sub.3): 8.39 (bs, 1H), 7.93 (s, 1H), 7.48-7.43 (m, 2H), 7.21 (s, 1H), 4.68 (t, J=6.84 Hz, 2H), 3.49 (t, J=6.84 Hz, 2H). Mass (MH): 214.0.

(196) Preparation of Intermediate AD

(197) ##STR00127##

(198) A suspension of Intermediate AC (2.5 g, 11.6 mmol), zinc powder (17.9 g) in methanol (330.0 mL) and 2N HCl (330.0 mL) was stirred at 85 C. for 2 hours. The reaction mixture was basified (pH10) and filtered. The cake was washed with methanol (310 mL), and the combined filtrate was concentrated under reduced pressure. The residue was dissolved in 5% MeOH in chloroform and washed with water. The organic layer was dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AD (1.5 g, 70%) as a brown gum. .sup.1H NMR (DMSO-d.sub.6): 11.43 (bs, 1H), 8.09 (s, 1H), 7.49 (d, J=8.49 Hz; 1H), 7.44-7.35 (m, 2H), 2.84-2.77 (m, 4H), 3.29 (t, J=7.03 Hz, 2H). Mass (M+H): 186.0.

(199) Preparation of Compound (19)

(200) ##STR00128##

(201) To a cold (0 C.) solution of Intermediate AD (300 mg, 1.62 mmol) and triethylamine (1.3 mL, 3.24 mmol) in dichloromethane (10.0 mL), was added slowly methoxyacetyl chloride (0.22 mL, 2.43 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 3 hours. The reaction mixture was diluted with dichloromethane (30 mL), washed with water (215 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and removal of the solvent yielded the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 2% MeOH in chloroform as the eluent to afford Compound (19) (60 mg, 15%) as a pale brown gum. .sup.1H NMR (DMSO-d.sub.6): 11.42 (bs, 1H), 8.11 (s, 1H), 7.89 (t, J=5.59 Hz; 1H), 7.49 (d, J=8.29 Hz; 1H), 7.42-7.37 (m, 2H), 3.76 (s, 2H), 3.40-3.34 (m, 2H), 3.32 (s, 3H), 2.87 (t, J=7.25 Hz; 2H). Mass (M+H): 258.0. IR (cm.sup.1): 3386, 3210, 2921, 2222, 1651, 1542, 1124, 639. HPLC purity (%): 94.72 (Max plot), 92.32 (254 nm), 94.91 (215 nm).

(202) Preparation of Compound (20)

(203) ##STR00129##

(204) To a cold (0 C.) solution of Compound (19) (50 mg, 0.194 mmol) and 3N NaOH (2.5 mL) in ethanol (3.5 mL) was added H.sub.2O.sub.2 (30% in water, 0.2 mL). After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 20 hours.

(205) The reaction mixture was concentrated, and the obtained aqueous residue was diluted with dichloromethane (50 mL), washed with water (210 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 8% MeOH in chloroform as the eluent to afford Compound (20) (36 mg, 33.6%) as a pale brown gum. .sup.1H NMR (DMSO-d.sub.6): 11.03 (bs, 1H), 8.17 (s, 1H), 7.83-7.78 (m, 2H), 7.63 (d, J=8.2 Hz; 1H), 7.32 (d, J=8.39 Hz; 1H), 7.22 (s, 1H), 7.06 (bs, 1H), 3.78 (s, 2H), 3.42-3.41 (m, 2H), 3.32 (s, 3H), 2.88 (t, J=6.93 Hz; 2H). Mass (M+H): 275.9. IR (cm.sup.1): 3433, 32923, 1645, 1239, 789. HPLC purity (%): 89.89 (Max plot), 95.47 (254 nm), 90.40 (215 nm).

(206) Synthesis of Compound (21)

(207) ##STR00130##

(208) Compound (21) was synthesized according to the procedure shown in Scheme 15.

(209) ##STR00131##

(210) Preparation of Intermediate AE

(211) ##STR00132##

(212) To a cold (0 C.) suspension of 60% NaH (0.51 g, 15.71 mmol) in DMF (10.0 mL), a solution of Intermediate K (3.0 g, 13.43 mmol) in DMF (10.0 mL) was added slowly over 5 minutes. The reaction stirred for 30 minutes; iodomethane (0.98 mL, 15.71 mmol) was then added, and the reaction stirred at room temperature for 1.5 hours. The reaction mixture was quenched with ice cold water (50 mL) and extracted with ethyl acetate (250 mL). The combined ethyl acetate layers were washed with water (220 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AE (520 mg, 96%), which was used in the next step without further purifications. .sup.1H NMR (CDCl.sub.3): 7.48-7.46 (m, 2H), 7.39-7.35 (m, 2H), 7.32-7.28 (m, 1H), 7.24-7.16 (m, 2H), 7.01-6.95 (m, 2H), 6.38 (d, J=2.92 Hz; 1H), 5.10 (s, 2H), 3.75 (s, 3H). Mass (M+H): 238.1.

(213) Preparation of Intermediate AF

(214) ##STR00133##

(215) POCl.sub.3 (0.21 mL, 2.32 mmol) was added to DMF (1.0 mL) dropwise at 0 C.-10 C. The resulting mixture was stirred for 30 minutes, cooled to 0 C., and a solution of Intermediate AE (0.5 g, 2.11 mmol) in DMF (1.0 mL) was added over 15 minutes. After the addition was complete, the reaction mixture was stirred at ambient temperature for 2 hours. The reaction mixture was quenched with ice (25 g), poured into water (20 mL), and basified (pH10) using 1N NaOH solution. The mixture was extracted with ethyl acetate (250 mL), and the combined ethyl acetate layers were washed with water (220 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and removal of the solvent afforded crude Intermediate AF, which was washed with petroleum ether (25 mL) and dried to afford the product (480 mg, 85%) as yellow solid. .sup.1H NMR (CDCl.sub.3): 9.95 (s, 1H), 7.91 (s, 1H), 7.62 (s, 1H), 7.50-7.48 (m, 2H), 7.48-7.30 (m, 3H), 7.26-7.24 (m, 2H), 7.08-7.05 (dd, 1H), 5.15 (s, 2H), 3.84 (s, 3H). Mass (M+H): 266.0.

(216) Preparation of Intermediate AG

(217) ##STR00134##

(218) A suspension of Intermediate AF (0.47 g, 1.77 mmol) and ammonium acetate (0.47 g, 6.09 mmol) in nitromethane (33.3 mL) was stirred at 90 C. for 1 hour. The reaction mixture was concentrated under reduced pressure, and the obtained crude product was washed with 25% ethyl acetate in petroleum ether (220 mL) and dried to afford Intermediate AG (0.48 g, 87%) as yellow solid. .sup.1H NMR (DMSO-d.sub.6): 8.23 (d, J=13.28 Hz; 1H), 7.65 (d, J=13.28 Hz; 1H), 7.51-7.24 (m, 8H), 7.09-7.07 (m, 1H), 5.16 (s, 2H), 3.83 (s, 3H). Mass (M+H): 309.0.

(219) Preparation of Intermediate AH

(220) ##STR00135##

(221) To a cold (0 C.) solution of Intermediate AG (0.7 g, 2.27 mmol) in methanol and DMF (2:1; 15 mL), NaBH.sub.4 (0.17 g, 4.54 mmol) was added portionwise over 20 minutes. After the addition was complete, the reaction mixture was stirred at 5 C. for 2 hours. The reaction mixture was quenched with water (10 mL) and concentrated under reduced pressure. The resulting aqueous residue was diluted with water (10 mL) and extracted with ethyl acetate (220 mL). The combined ethyl acetate layers were washed with water (10 mL) and brine solution (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product, which was purified by column chromatography (silica gel 100-200 mesh) using 20% ethyl acetate in petroleum ether as the eluent to afford Intermediate AH (440 mg, 62%) as a pale yellow solid.

(222) .sup.1H NMR (CDCl.sub.3): 7.49-7.47 (m, 2H), 7.41-7.32 (m, 3H), 7.20 (d, J=9.12 Hz; 1H), 7.06-7.05 (s, 1H), 7.00-6.97 (m, 1H), 6.88 (s, 1H), 5.11 (s, 2H), 4.60 (t, J=7.25 Hz; 2H), 3.71 (s, 3H), 3.42 (t, J=7.25 Hz; 2H). Mass (M+H): 311.1.

(223) Preparation of Intermediate AI

(224) ##STR00136##

(225) A suspension of Intermediate AH (430 mg, 1.381 mmol) and zinc powder (2.13 mg, 32.59 mmol) in methanol (57.0 mL) and 2N HCl (57.0 mL) was stirred at 65 C. for 2 hours. The reaction mixture was basified (pH10) and filtered. The cake was washed with methanol (310 mL), and the combined filtrate was concentrated under reduced pressure. The residue was dissolved in 5% MeOH in chloroform (150 mL) and washed with water (20 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AI (160 mg, 41%) as a brown gum. .sup.1H NMR (DMSO-d.sub.6): 7.48-7.25 (m, 6H), 7.12-7.06 (m, 2H), 6.86-6.83 (m, 1H), 5.09 (s, 2H), 3.68 (s, 3H), 2.79-2.68 (m, 4H). Mass (M+H): 281.1.

(226) Preparation of Intermediate AJ

(227) ##STR00137##

(228) To a cold (0 C.) solution of Intermediate AI (150 mg, 0.535 mmol) and triethylamine (0.15 mL, 1.07 mmol) in dichloromethane (10.0 mL) was added slowly a solution of methoxyacetyl chloride (0.06 mL, 0.64 mmol) in dichloromethane (2.0 mL) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (220 mL). The combined dichloromethane layer was washed with water (10 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product, which was purified by column chromatography (silica gel 100-200 mesh) using 2% MeOH in chloroform as the eluent to afford Intermediate AJ (85 mg, 45%) as a brown gum. .sup.1H NMR (CDCl.sub.3): 7.49-7.47 (m, 2H), 7.41-7.30 (m, 3H), 7.21-7.13 (m, 2H), 6.99-6.96 (m, 1H), 6.87 (s, 1H), 6.63 (bs, 1H), 5.11 (s, 2H), 3.87 (s, 2H), 3.72 (s, 3H), 3.64-3.57 (m, 2H), 3.33 (s, 3H), 2.93 (t, J=6.84 Hz; 2H). Mass (M+H): 353.1.

(229) Preparation of Compound (21)

(230) ##STR00138##

(231) A suspension of Intermediate AJ (80 mg, 0.227 mmol) and 10% Pd/C (40 mg, dry) in MeOH (10.0 mL) was hydrogenated (40 psi H.sub.2 pressure) at room temperature for 2 hours. The reaction mixture was filtered through a Celite bed, and the cake was washed with methanol (35 mL). The combined filtrates were concentrated under reduced pressure to give the crude product, which was purified by column chromatography (silica gel 100-200 mesh) using 2% MeOH in chloroform as the eluent to afford Compound (21) (30 mg, 50%) as a brown solid. .sup.1H NMR (CDCl.sub.3): 7.15 (d, J=8.85 Hz; 1H), 7.02 (s, 1H), 6.85-6.80 (m, 2H), 6.66 (bs, 1H), 5.02 (s, 1H), 3.88 (s, 2H), 3.71 (s, 3H), 3.61-3.56 (m, 2H), 3.33 (s, 3H), 2.91 (t, J=6.97 Hz; 2H). Mass (M+H): 262.9. IR (cm.sup.1): 3400, 2926, 1651, 1219, 1114, 771. HPLC purity (%): 93.49 (Max plot), 94.77 (254 nm), 97.19 (215 nm).

(232) Synthesis of Compounds (22) and (23)

(233) ##STR00139##

(234) Compounds (22) and (23) were prepared according to the procedure shown in Scheme 16.

(235) ##STR00140##

(236) Preparation of Intermediate AL

(237) ##STR00141##

(238) To a cold (0 C.) suspension of 60% NaH (0.58 g, 14.51 mmol) in DMF (10.0 mL) was added slowly a solution of Intermediate AK (2.0 g, 12.40 mmol) in DMF (5.0 mL) was added slowly for 5 minutes. The reaction was then stirred for 30 minutes, and iodomethane (2.06 g, 14.52 mmol) was then added to the reaction mixture. The reaction stirred at room temperature for 5 hours. The reaction mixture was quenched with ice cold water (25 mL) and extracted with ethyl acetate (330 mL). The combined ethyl acetate layers were washed with water (220 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AL which was purified by column chromatography (silica gel 100-200 mesh) using 12% ethyl acetate in petroleum ether as the eluent to afford the product (1.25 g, 57%) as a brown solid. .sup.1H NMR (CDCl.sub.3): 7.13 (d, J=8.78 Hz; 1H), 7.00 (s, 1H), 6.81-6.78 (dd, 1H), 6.16 (s, 1H), 3.83 (s, 3H), 3.62 (s, 3H), 2.39 (s, 3H). Mass (M+H): 176.0.

(239) Preparation of Intermediates AM1 and AM2

(240) ##STR00142##

(241) General Procedure.

(242) POCl.sub.3 (1.1 mmol) was added to DMF (2.0 mL) dropwise at 0 C.-10 C. The resulting mixture was stirred for 30 minutes, cooled to 0 C., and a solution of Intermediate AK or Intermediate AL (1.0 mmol) in DMF (2.0 mL) was added over 15 minutes. After the addition was complete, the reaction mixture was stirred at ambient temperature for 2 hours. The reaction mixture was quenched with ice (25 g), poured into water (30 mL), and basified (pH10) using a 1N NaOH solution. The mixture was extracted using ethyl acetate (320 mL), washed with water (210 mL) and brine (15 mL), and dried over anhydrous Na.sub.2SO.sub.4, and removal of the solvent afforded Intermediate AM1 or AM2 (Table 10).

(243) TABLE-US-00010 TABLE 10 AM1 Intermediate AK (2.0 g, 12.4 mmol) was reacted with POCl.sub.3 (1.67 mL, 13.64 mmol) in DMF (8.0 mL) to give Inter- mediate AM1 (1.7 g, 72%) as a pale brown solid. .sup.1H NMR (CDCl.sub.3): 10.15 (s, 1H), 8.35 (bs, 1H), 7.77 (s, 1H), 7.21 (d, J = 8.78 Hz; 1H), 6.88 (dd, 1H), 3.88 (s, 3H), 2.72 (s, 3H). Mass (M + H): 189.9. AM2 Intermediate AL (1.25 g, 7.14 mmol) was reacted with POCl.sub.3 (0.74 mL, 7.85 mmol) in DMF (10.0 mL) to give Intermediate AM2 (1.2 g, 82%) as a pale brown solid. .sup.1H NMR (CDCl.sub.3): 10.12 (s, 1H), 7.80 (s, 1H), 7.19 (d, J = 8.85 Hz; 1H), 6.91 (dd, 1H), 3.89 (s, 3H), 3.68 (s, 3H), 2.66 (s, 3H), Mass (M + H): 203.9.

(244) Preparation of Intermediates AN1 and AN2

(245) ##STR00145##

(246) A suspension of Intermediate AM1 or AM2 (1.0 mmol), ammonium acetate (3.43 mmol) in nitromethane (80 mL) was stirred at 90 C. for 7 hours. The reaction mixture was concentrated under reduced pressure. The resulting crude product was dissolved in ethyl acetate (100 mL), washed with water (230 mL) and brine solution (25 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AN (Table 11).

(247) TABLE-US-00011 TABLE 11 AN1 Intermediate AM1 (1.7 g, 8.99 mmol) was reacted with ammonium acetate (2.3 g, 30.85 mmol) in nitromethane (120.0 mL) to give Intermediate AN1 (2.1 g, 98%) as a yellow solid. .sup.1H NMR (CDCl.sub.3): 8.52 (bs, 1H), 8.33 (d, J = 13.42 Hz; 1H), 7.71 (d, J = 13.42 Hz; 1H), 7.27-7.25 (m, 1H), 7.11 (d, 1H), 6.91-6.89 (m, 1H), 3.90 (s, 3H), 2.62 (s, 3H). Mass (M + H): 203.9. AN2 Intermediate AM2 (1.2 g, 5.91 mmol) was reacted with ammonium acetate (1.56 g, 20.27 mmol) in nitromethane (85.0 mL) to give Intermediate AN2 (1.4 g, 96%) as a pale yellow solid. .sup.1H NMR (CDCl.sub.3): 8.33 (d, J = 13.15 Hz; 1H), 7.71 (d, J = 13.15 Hz; 1H), 7.26-7.23 (m, 1H), 7.12 (s, 1H), 6.95-6.92 (m, 1H), 3.90 (s, 3H), 3.72 (s, 3H), 2.62 (s, 3H). Mass (M + H): 246.9.

(248) Preparation of Intermediates AO1 and AO2

(249) ##STR00148##

(250) To a cold (0 C.) solution of Intermediate AN1 or AN2 (1.0 mmol) in methanol and DMF (1:1; 35 mL), was added portionwise NaBH.sub.4 (2.0 mmol) over 20 minutes. After the addition was complete, the reaction mixture was allowed to stir at 10 C. for 2 hours. The reaction mixture was quenched with water (10 mL) and concentrated under reduced pressure. The resulting aqueous residue was diluted with water (20 mL) and extracted with ethyl acetate (230 mL). The combined ethyl acetate layers were washed with water (220 mL) and brine solution (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AO (Table 12).

(251) TABLE-US-00012 TABLE 12 AO1 Intermediate AN1 (1.0 g, 4.74 mmol) was reacted with NaBH.sub.4 (360 mg, 9.48 mmol) in MeOH (40.0 mL) to give Intermediate AO1 (1.1 g, crude) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.75 (bs, 1H), 7.17 (d, J = 8.70 Hz; 1H), 6.90 (s, 1H), 6.81-6.78 (m, 1H), 4.57 (t, J = 7.25 Hz; 2H), 3.86 (s, 3H), 3.40 (t, J = 7.46 Hz, 2H), 2.37 (s, 3H). Mass (M + H): 235.1. AO2 0 Intermediate AN2 (1.3 g, 5.28 mmol) was reacted with NaBH.sub.4 (400 mg, 10.56 mmol) in MeOH (45.0 mL) to give Intermediate AO2 (1.4 g, crude) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.15 (d, J = 8.70 Hz; 1H), 6.91 (s, 1H), 6.85-6.84 (m, 1H), 4.55 (t, J = 7.46 Hz; 2H), 3.86 (s, 3H), 3.62 (s, 3H), 3.42 (t, J = 7.46 Hz, 2H), 2.34 (s, 3H). Mass (M + H): 249.1.

(252) Preparation of Intermediate AP1 and AP2

(253) ##STR00151##

(254) To a cold (70 C.) solution of Intermediate AO (1.0 mmol) in dichloromethane (30 mL) was added slowly BBr.sub.3 (2.0 mmol). After the addition was complete, the reaction mixture was allowed to reach 0 C. and stirred for 4 hours. The reaction mixture was diluted with dichloromethane (25 mL), washed with water (210 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AP, each of which was purified by column chromatography (silica gel 100-200 mesh) using 20% ethyl acetate in petroleum ether as the eluent (Table 13).

(255) TABLE-US-00013 TABLE 13 AP1 Intermediate AO1 (600 mg, 2.56 mmol) was reacted with BBr.sub.3 (0.74 mL, 5.12 mmol) in dichloromethane (20.0 mL) to give Intermediate AP1 (100 mg, 18%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.75 (bs, 1H), 7.12 (d, J = 8.78 Hz; 1H), 6.86 (s, 1H), 6.71-6.68 (m, 1H), 4.73 (bs, 1H), 4.54 (t, J = 7.31 Hz; 2H), 3.35 (t, J = 7.31 Hz, 2H), 2.36 (s, 3H). Mass (M + H): 221.0. AP2 Intermediate AO2 (600 mg, 2.41 mmol) was reacted with BBr.sub.3 (0.47 mL, 4.84 mmol) in dichloromethane (20.0 mL) to give Intermediate AP2 (75 mg, 13%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.11 (d, J = 8.70 Hz; 1H), 6.89 (s, 1H), 6.75- 6.72 (m, 1H), 4.56-4.50 (m, 2H), 3.61 (s, 3H), 3.37 (t, J = 7.46 Hz, 2H), 2.34 (s, 3H).

(256) Preparation of Intermediates AQ1 and AQ2

(257) ##STR00154##

(258) A suspension of Intermediate AP (1.0 mmol) and 10% Pd/C (60% w/w, dry) in MeOH (30 mL) was hydrogenated (40 psi H.sub.2 pressure) at 26 C. for 2 hours. The reaction mixture was filtered, the cake was washed with methanol (35 mL), and the combined filtrates were concentrated under reduced pressure to afford Intermediates AQ (Table 14), which was used as such in the next step.

(259) TABLE-US-00014 TABLE 14 AQ1 Intermediate AP1 (100 mg, 0.454 mmol) was hydrogenated with 10% Pd/C (60 mg) in MeOH (20.0 mL) to give Intermediate AQ1 (80 mg, crude) as a pale brown gum. Mass (M + H): 191.1. AQ2 Intermediate AP2 (125 mg, 0.53 mmol) was hydrogenated with 10% Pd/C (70 mg) in MeOH (20.0 mL) to give Intermediate AQ2 (108 mg, crude) as a pale brown gum. Mass (M + H): 205.1.

(260) Preparation of Intermediates AR1 and AR2

(261) ##STR00157##

(262) To a cold (0 C.) solution of crude Intermediate AQ (1.0 mmol) and triethylamine (2.2 mmol) in dichloromethane (20 mL), methoxyacetyl chloride (2.2 mmol) was added slowly over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (25 mL), washed with water (210 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AR (Table 15), which was purified by column chromatography (silica gel 100-200 mesh) using 20% ethyl acetate in petroleum ether as the eluent.

(263) TABLE-US-00015 TABLE 15 AR1 Intermediate AQ1 (80 mg, 0.42 mmol) was reacted with methoxyacetyl chloride (0.04 mL, 0.42 mmol) and Et.sub.3N (0.06 mL, 0.46 mmol) in dichloro- methane (15.0 mL) to give Intermediate AR1 (46 mg, 33%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.87 (bs, 1H), 7.23-7.21 (m, 2H), 6.87- 6.84 (m, 1H), 6.58 (bs, 1H), 4.31 (s, 2H), 3.85 (s, 2H), 3.56 (s, 3H), 3.53-3.48 (q, 2H), 3.31 (s, 3H), 2.90-2.86 (m, 2H), 2.38 (s, 3H). Mass (M + H): 335.1. AR2 Intermediate AQ2 (108 mg, 0.53 mmol) was reacted with methoxy- acetyl chloride (0.11 mL, 1.16 mmol) and Et.sub.3N (0.18 mL, 1.164 mmol) in dichloro- methane (15.0 mL) to give Intermediate AR2 (40 mg, 22%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.23- 7.21 (m, 2H), 6.91- 6.88 (m, 1H), 6.76 (bs, 1H), 4.31 (s, 2H), 3.85 (s, 2H), 3.66 (s, 3H), 3.51-3.46 (m, 2H), 3.31 (s, 3H), 2.91 (t, J = 7.05 Hz; 2H), 2.36 (s, 3H). Mass (M + H): 363.1.

(264) Preparation of Compounds (22) and (23)

(265) ##STR00160##

(266) A suspension of Intermediate AR (1.0 mmol) and K.sub.2CO.sub.3 (1.1 mmol) in methanol (20 mL) was stirred at 26 C. for 2 hours. The reaction mixture was concentrated and the residue was diluted with water (20 mL) and extracted with ethyl acetate (220 mL). The combined ethyl acetate layers were washed with brine (210 mL), dried over anhydrous Na.sub.2SO.sub.4, and concentrated to afford the crude product, which was purified by PREP-TLC using 70% ethyl acetate in petroleum ether as the eluent to afford the corresponding Compounds (22) and (23) (Table 16).

(267) TABLE-US-00016 TABLE 16 (22) Intermediate AR1 (46 mg, 0.14 mmol) was reacted with K.sub.2CO.sub.3 (20 mg, 0.15 mmol) in MeOH (3.0 mL) to give Compound (22) (15 mg, 36%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.65 (bs, 1H), 7.13 (d, J = 8.70 Hz; 1H), 6.94 (s, 1H), 6.70-6.68 (m, 1H), 6.62 (bs, 1H), 4.79 (bs, 1H), 3.86 (s, 2H), 3.54-3.49 (m, 2H), 3.32 (s, 3H), 2.86 (t, J = 6.84 Hz; 2H), 2.35 (s, 3H). Mass (M + H): 263.1. IR (cm.sup.1): 3382, 2923, 1649, 1200, 1114, 799. HPLC purity (%): 97.01 (Max plot), 95.28 (254 nm), 97.22 (215 nm). (23) Intermediate AR2 (40 mg, 0.114 mmol) was reacted with K.sub.2CO.sub.3 (17 mg, 0.126 mmol) in MeOH (3.0 mL) to give Compound (23) (14 mg, 38%) Pale brown solid. .sup.1H NMR (CDCl.sub.3): 7.10 (d, J = 8.78 Hz; 1H), 6.96 (s, 1H), 6.75-6.72 (m, 1H), 6.65 (bs, 1H), 4.93 (bs, 1H), 3.86 (s, 2H), 3.62 (s, 3H), 3.51-3.46 (m, 2H), 3.31 (s, 3H), 2.89 (t, J = 6.83 Hz; 2H), 2.33 (s, 3H). Mass (M + H): 277.1. IR (cm.sup.1): 3378, 2925, 1661, 1211, 1116, 795. HPLC purity (%): 93.05 (Max plot), 80.02 (254 nm), 93.42 (215 nm).
Synthesis of Compound (24)

(268) ##STR00163##

(269) Compound (24) was synthesized according to the procedure shown in Scheme 17.

(270) ##STR00164##

(271) Preparation of Intermediate AS

(272) ##STR00165##

(273) A solution of Intermediate K (3.0 g, 13.45 mmol) and pyridine (1.8 mL, 22.86 mmol) in dioxane (25.0 mL) was stirred at 65 C. for 1 hour. A solution of chloroacetyl chloride (1.8 mL, 22.86 mmol) in dioxane (5.0 mL) was then added dropwise. After the addition was complete, the reaction mixture was stirred for 1 hour. The reaction mixture was cooled, poured into cold ether (150 mL), and stirred. The resulting solid was filtered, washed with cold ether (220 mL), and dried to afford crude Intermediate AS (1.6 g, 40%) as a yellow solid. .sup.1H NMR (DMSO-d.sub.6): 12.04 (bs, 1H), 8.38 (s, 1H), 7.76 (s, 1H), 7.49-7.32 (m, 6H), 6.96 (m, 1H), 5.13 (s, 2H), 4.84 (s, 2H). Mass (M+H): 300.0.

(274) Preparation of Intermediate AT

(275) ##STR00166##

(276) To a solution of Intermediate AS (1.6 g, 5.35 mmol) in acetone (80.0 mL) and water (40.0 mL), was added NaN.sub.3 (800 mg, 1.23 mmol) and the resulting reaction mixture was stirred at 80 C. for 16 hours. The reaction mixture was cooled to room temperature, diluted with water (100 mL), and extracted with dichloromethane (2100 mL). The combined organic layers were washed with water (50 mL) and brine (30 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to yield the crude Intermediate AT. This material was washed with petroleum ether (210 mL) and dried to afford Intermediate AT (12 g, 73%) as a yellow solid.

(277) .sup.1H NMR (CDCl.sub.3): 8.52 (bs, 1H), 7.99 (s, 1H), 7.85 (s, 1H), 7.50 (d, J=7.31 Hz; 1H), 7.41-7.32 (m, 4H), 7.05 (m, 1H), 5.16 (s, 2H), 4.37 (s, 2H). Mass (MH): 305.0. IR (cm.sup.1): 3190, 2924, 2103, 1641, 1259, 746.

(278) Preparation of Intermediate AU

(279) ##STR00167##

(280) A suspension of Intermediate AT (1.2 g, 3.93 mmol) and 10% Pd/C (750 mg, dry) in MeOH (40.0 mL) was hydrogenated (60 psi H.sub.2 pressure) at room temperature for 3 hours. The reaction mixture was filtered, and the cake was washed with methanol (35 mL). The combined filtrates were concentrated under reduced pressure to afford Intermediate AU (750 mg, crude) as a brown solid, which was used without further purification in the next step. Mass (M+H): 191.0.

(281) Preparation of Intermediate AV

(282) ##STR00168##

(283) To a cold (0 C.) solution of Intermediate AU (500 mg, 2.63 mmol) and triethylamine (1.09 mL, 7.89 mmol) in dichloromethane (20.0 mL) was added slowly methoxyacetyl chloride (430 mg, 3.94 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (50 mL), washed with water (220 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AV (800 mg, crude) which was used in next step without further purification.

(284) Preparation of Compound (24)

(285) ##STR00169##

(286) A suspension of crude Intermediate AV (800 mg, 2.63 mmol) and K.sub.2CO.sub.3 (363 mg, 2.63 mmol) in methanol (15.0 mL) was stirred at 26 C. for 1 hour. The reaction mixture was concentrated under reduced pressure; the residue was diluted with water (20 mL) and extracted with ethyl acetate (220 mL). The combined ethyl acetate layers were washed with water (10 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (100-200 mesh silica gel) using 10% MeOH in chloroform as the eluent to afford Compound (24) (190 mg, 30%) as a pale yellow solid. .sup.1H NMR (DMSO-d.sub.6): 11.75 (bs, 1H), 9.01 (s, 1H), 8.30 (s, 11H), 7.92 (m, 1H), 7.54 (s, 1H), 7.26 (d, J=8.70 Hz 1H), 6.71 (m, 1H), 4.47 (d, J=5.39 Hz; 1H), 3.89 (s, 2H), 3.38 (s, 3H). Mass (M+H): 263.0. IR (cm.sup.1): 3259, 2930, 1661, 1614, 1215, 1122, 924. HPLC purity (%): 93.7 (Max plot), 96.40 (254 nm), 94.59 (215 nm).

(287) Synthesis of Compound (25)

(288) ##STR00170##

(289) Compound (25) can be synthesized according to the procedure shown in Scheme 18. The aniline starting material can be transformed to the corresponding arylhydrazine. Treatment of this arylhydrazine intermediate with 4-chlorobutyraldehyde diethyl acetal can afford the requisite indole intermediate. N-acylation followed by reduction of the C5 ester can result in the desired Compound (25).

(290) ##STR00171##
Synthesis of Formula (II) Compounds
Synthesis of Compound (26)

(291) ##STR00172##

(292) Compound (26) was synthesized according to the procedure shown in Scheme 19.

(293) ##STR00173##

(294) To a cold (0 C.) solution of Intermediate AW (250 mg, 1.79 mmol) and triethylamine (0.3 mL, 2.13 mmol) in dichloromethane (5.0 mL) was added slowly methoxyacetyl chloride (0.18 mL, 2.12 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 3 hours. The reaction mixture was diluted with dichloromethane (20 mL), washed with water (220 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product, which was purified by column chromatography (silica gel 100-200 mesh) using 2% MeOH in chloroform as the eluent to afford Compound (26) (380 mg, 88%) as a pale brown oil. .sup.1H NMR (DMSO-d.sub.6): 7.30-7.24 (m, 1H), 6.99-6.90 (m, 3H), 6.58 (bs, 1H), 3.87 (s, 21H), 3.55 (q, J=13.26 Hz; J=7.09 Hz; 2H), 3.36 (s, 3H), 2.84 (t, J=7.04, Hz; 2H). Mass (M+H): 212.0. IR (cm.sup.1): 3418, 2934, 1671, 1534, 1116, 783. HPLC purity (%): 97.37 (Max plot), 98.95 (215 nm).

(295) Synthesis of Compound (27)

(296) ##STR00174##

(297) Compound (27) was prepared according to the procedure shown in Scheme 20.

(298) ##STR00175##

(299) Preparation of Intermediate AY

(300) ##STR00176##

(301) To a solution of Intermediate AX (1.0 g, 4.99 mmol) in THF (20 mL) at 0 C. was added a solution of BH.sub.3DMS (0.94 mL, 9.98 mmol). After the addition was complete, the reaction mixture was stirred at 70 C. for 1 hour. The reaction mixture was cooled, methanol (5.0 mL) was added, and the mixture was refluxed for 30 minutes. Solvent from the reaction mixture was removed via distillation, and the residue was diluted with ethyl acetate (30 mL), washed with water (215 mL) and brine (15 mL), dried over dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AY (720 mg, 77%) as an off white solid. This material was used in the next step without further purifications. .sup.1H NMR (CDCl.sub.3): 7.95 (s, 1H), 7.85 (d, J=7.88 Hz; 1H), 7.66 (d, J=7.88 Hz, 1H), 7.56 (t, J=7.65 Hz; 1H), 4.80 (s, 2H), 3.06 (s, 3H). Mass (M+H): 187.0.

(302) Preparation of Intermediate AZ

(303) ##STR00177##

(304) To a cold (0 C.) solution of Intermediate AY (2 g, 10.75 mmol) and triethylamine (2.26 mL, 16.12 mmol) in dichloromethane (25 mL) was added slowly methanesulfonyl chloride (1.08 mL, 13.85 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 16 hours. The reaction mixture was quenched with cold water (10 mL), diluted with dichloromethane (20 mL), washed with cold water (250 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate AZ. This material was purified by column chromatography (silica gel 100-200 mesh) using 10% ethyl acetate in petroleum ether as the eluent to afford the product (1.6 g, 73%) as a pale yellow oil. .sup.1H NMR (CDCl.sub.3): 7.98 (s, 1H), 7.91 (d, J=7.80 Hz; 1H), 7.70 (d, J=7.80 Hz, 1H), 7.59 (t, J=7.80 Hz; 1H), 4.65 (s, 2H), 3.07 (s, 3H). Mass (M+H): 205.0.

(305) Preparation of Intermediate BA

(306) ##STR00178##

(307) To a cold solution of Intermediate AZ (500 mg, 2.45 mmol) in DMSO (5.0 mL) at 0 C. was added sodium cyanide (240 mg, 4.89 mmol) portionwise over 15 minutes. After the addition was complete, the reaction mixture was allowed and stirred at 10 C. for 1 hour. Ice cold water (20 mL) was added to the reaction mixture. The reaction was then extracted with ethyl acetate (320 mL), washed with water (20 mL) and brine (15 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate BA. This material was purified by column chromatography (silica gel 100-200 mesh) using 40% ethyl acetate in petroleum ether as the eluent to afford the product (280 mg, 59%) as a pale brown solid. .sup.1H NMR (CDCl.sub.3): 7.95-7.91 (m, 2H), 7.69-7.61 (m, 2H), 3.86 (s, 2H), 3.07 (s, 3H). Mass (MH): 194.0. IR (cm.sup.1): 2927, 2249, 1300, 1144, 964, 761.

(308) Preparation of Compound (27)

(309) ##STR00179##

(310) A suspension of Intermediate BA (250 mg, 1.28 mmol) and Raney-Ni (100 mg, wet) in methanolic NH.sub.3 (5.0 mL) was hydrogenated by bubbling H.sub.2 at 10 C. for 3 hours. The reaction mixture was filtered, the cake was washed with methanol (310 mL), and the combined filtrate was concentrated under reduced pressure. The resulting oily residue was dissolved in ethyl acetate (2.0 mL), cooled in ice, treated with EtOAc and HCl, and stirred for 10 minutes. The resulting solid was filtered, washed with ethyl acetate (35 mL), and dried to afford Compound (27).HCl (150 mg, 50%) as an off white solid. .sup.1H NMR (DMSO-d.sub.6): 7.91 (bs, 2H), 7.83-7.81 (m, 2H), 7.64-7.62 (m, 2H), 3.21 (s, 3H), 3.11-3.01 (m, 2H), 2.99-2.97 (m, 2H). Mass (M+H): 200.0. IR (cm.sup.1): 3402, 3034, 1601, 1290, 1141, 965, 767, 532. HPLC purity (%): 99.92 (Max plot), 99.90 (215 nm).

(311) Preparation of Compound (28)

(312) ##STR00180##

(313) Compound (28) was prepared according to the procedure shown in Scheme 21.

(314) ##STR00181##

(315) Preparation of Intermediate BC

(316) ##STR00182##

(317) To a cold solution of Intermediate BB (2 g, 9.25 mmol) in DMSO (20.0 mL) at 0 C. was added sodium cyanide (900 mg, 18.36 mmol) portionwise over 15 minutes. After the addition was complete, the reaction mixture was allowed and stirred at 10 C. for 3 hours. Ice cold water was added to the reaction mixture (30 mL), and the reaction mixture was extracted with ethyl acetate (325 mL), washed with water (20 mL) and brine (15 mL), dried over dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. The mixture was purified by column chromatography (silica gel 100-200 mesh) using 40% ethyl acetate in petroleum ether as the eluent to afford Intermediate BC (800 mg, 53%) as a brown gum. .sup.1H NMR (CDCl.sub.3): 8.23-8.21 (m, 2H), 7.72 (d, J=7.80 Hz, 1H), 7.62 (m, 1H), 3.89 (s, 2H). Mass (MH): 161.0.

(318) Preparation of Intermediate BD

(319) ##STR00183##

(320) To a stirred solution of NH.sub.4Cl (1.1 g, 19.74 mmol) in H.sub.2O (16 mL) was added Fe powder (1.01 g, 18.08 mmol) followed by Intermediate BC (800 mg, 4.23 mmol) in a mixture of THF (8.0 mL) and MeOH (8.0 mL) slowly at room temperature. The reaction was then stirred for 3 h at 60 C. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (100 mL), and filtered through a Celite bed. The organic layer was washed with water (225 mL) and brine solution (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and evaporated to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 40% ethyl acetate in petroleum ether as the eluent to afford Intermediate BD (520 mg, 80%) as a brown gum. .sup.1H NMR (CDCl.sub.3): 7.14 (d, J=7.67 Hz, 1H), 7.68-6.62 (m, 3H), 3.74 (bs, 2H), 3.65 (s, 2H). Mass (M+H): 238.0.

(321) Preparation of Intermediate BE

(322) ##STR00184##

(323) To a cold (0 C.) solution of Intermediate BD (500 mg, 3.78 mmol) and triethylamine (0.63 mL, 4.48 mmol) in dichloromethane (10 mL) was added slowly methanesulfonyl chloride (035 mL, 4.49 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (10 mL), washed with water (220 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 15% ethyl acetate in petroleum ether as the eluent to afford Intermediate BE (300 mg, 38%) as a brown gum. .sup.1H NMR (CDCl.sub.3): 7.40-7.34 (m, 1H), 7.20-7.16 (m, 2H), 6.63 (bs, 1H), 3.76 (s, 2H), 3.04 (s, 3H). Mass (MH): 187.0.

(324) Preparation of Intermediate BF

(325) ##STR00185##

(326) A suspension of Intermediate BE (100 mg, 0.47 mmol) and Raney-Ni (20 mg, wet) in methanolic NH.sub.3 (5.0 mL) was hydrogenated by bubbling H.sub.2 at 10 C. for 3 hours. The reaction mixture was filtered, and the cake was washed with methanol (310 mL). The combined filtrates were concentrated under reduced pressure to afford Intermediate BF (80 mg, crude) as a pale brown oil, which was used in the next step without further purification.

(327) Preparation of Compound (28)

(328) ##STR00186##

(329) To a cold (0 C.) solution of crude Intermediate BF (30 mg, 0.14 mmol) and triethylamine (0.02 mL, 0.14 mmol) in dichloromethane (4.0 mL) was added slowly a solution of methoxyacetyl chloride (0.03 mL, 0.13 mmol) in dichloromethane (1.0 mL) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (10 mL), washed with water (210 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 23% ethyl acetate in petroleum ether as the eluent to afford Compound (28) (29 mg, 72%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.31-7.25 (m, 2H), 7.09-7.03 (m, 2H), 6.58 (bs, 1H), 6.50 (bs, 1H), 3.87 (s, 2H), 3.59-3.53 (q, 2H), 3.37 (s, 3H), 2.84 (t, J=7.11 Hz; 2H). Mass (M+H): 286.9. IR (cm.sup.1): 3401, 2927, 1655, 1328, 1149, 976, 763, 515. HPLC purity (%): 94.51 (Max plot), 92.11 (215 nm).

(330) Synthesis of Compounds (29) and (30)

(331) ##STR00187##

(332) Compounds (29) and (30) were prepared according to the procedure in Scheme 22.

(333) ##STR00188##

(334) Preparation of Intermediate BH

(335) ##STR00189##

(336) A suspension of Intermediate BG (5 g, 27.02 mmol) and ammonium acetate (4.57 g, 59.45 mmol) in nitromethane (150.0 mL) was stirred at 90 C. for 1 hour. The reaction mixture was concentrated under reduced pressure, and the resulting crude material was dissolved in ethyl acetate (100 mL), washed with water (230 mL) and brine solution (25 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 2% ethyl acetate in petroleum ether as the eluent to afford Intermediate BH (2.2 g, 36%) as a yellow solid. .sup.1H NMR (CDCl.sub.3): 7.93 (d, J=13.66 Hz, 1H), 7.70 (s, 1H), 7.62 (d, J=8.00 Hz, 1H), 7.56 (d, J=13.66 Hz, 1H), 7.48 (d, J=7.61 Hz, 1H), 7.34 (s, J=7.90 Hz, 1H). Mass (M2H), M: 226.9, 228.9.

(337) Preparation of Intermediate BI

(338) ##STR00190##

(339) To a cold (0 C.) solution of Intermediate BH (2 g, 8.77 mmol) in methanol (30 mL) at 0 C. was added NaBH.sub.4 (0.4 g, 10.52 mmol) portionwise over 15 minutes. After the addition was complete, the reaction mixture was allowed to stir at 10 C. for 1 hour. The reaction mixture was quenched with water (10 mL) and concentrated under reduced pressure. The resulting aqueous residue was diluted with water (25 mL) and extracted with ethyl acetate (250 mL). The combined ethyl acetate layers were washed with water (30 mL) and brine solution (25 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 5% ethyl acetate in petroleum ether as the eluent to afford Intermediate BI (1.5 g, 74%) as pale yellow solid. .sup.1H NMR (CDCl.sub.3): 7.39 (d, J=10.78 Hz, 1H), 7.37 (s, 1H), 7.22-7.15 (m, 2H), 4.61 (t, J=7.46 Hz, 2H), 3.29 (t, J=7.03 Hz, 2H). Mass (M2H), M: 227.9, 229.9.

(340) Preparation of Intermediate BJ

(341) ##STR00191##

(342) A suspension of Intermediate BI (350 mg, 15.21 mmol) and zinc powder (300 mg, 4.56 mmol) in methanol (50.0 mL) and 2N HCl (50.0 mL) was stirred at 65 C. for 1 hour. The reaction mixture was filtered, and the cake was washed with methanol (310 mL). The combined filtrates were concentrated under reduced pressure, and the residue was dissolved in dichloromethane, dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate BJ (1 g, crude), which was used in the next step without further purification.

(343) Preparation of Intermediate BK

(344) ##STR00192##

(345) To a cold (0 C.) solution of crude Intermediate BJ (300 mg, 1.50 mmol) and triethylamine (0.42 mL, 3.01 mmol) in dichloromethane (15.0 mL) was added slowly a solution of methoxyacetyl chloride (0.15 mL, 1.65 mmol) in dichloromethane (2.0 mL) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (10 mL), washed with water (220 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 1% MeOH in chloroform as the eluent to afford Intermediate BK (200 mg, 60%) as a pale brown solid. .sup.1H NMR (CDCl.sub.3): 7.38-7.36 (m, 2H), 7.20-7.12 (m, 2H), 6.55 (bs, 1H), 3.87 (s, 2H), 3.56-3.51 (m, 2H), 3.37 (s, 3H), 2.81 (t, J=7.12 Hz, 2H). Mass (M, M+H): 271.9, 273.9.

(346) Preparation of Compound (29)

(347) ##STR00193##

(348) A suspension of Intermediate BK (300 mg, 1.10 mmol) and CuCN (200 mg, 2.20 mmol) in DMSO (4.0 mL) was stirred in a sealed tube at 165 C. for 20 hours. The reaction mixture was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (220 mL). The combined ethyl acetate layers were washed with water (210 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 2% MeOH in chloroform as the eluent to afford Compound (29) (100 mg, 41.5%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.55-7.42 (m, 4H), 6.6 (bs, 1H), 3.88 (s, 2H), 3.58-3.53 (q, 2H), 3.38 (s, 3H), 2.89 (t, J=7.31 Hz; 2H). Mass (MH): 217. IR (cm.sup.1): 3412, 2928, 2230, 1665, 1537, 1116, 798, 691. HPLC purity (%): 95.33 (Max plot), 92.55 (215 nm).

(349) Preparation of Compound (30)

(350) ##STR00194##

(351) To a cold (0 C.) solution of Compound (29) (100 mg, 0.45 mmol), 3N NaOH (3.0 mL) in ethanol (3.0 mL) at 0 C. was added H.sub.2O.sub.2 (30% in water, 0.05 mL). After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 20 hours. The reaction mixture was concentrated, and the resulting aqueous residue was diluted with dichloromethane (50 mL), washed with water (210 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 5% MeOH in chloroform as the eluent to afford Compound (30) (40 mg, 37%) as an off white solid. .sup.1H NMR (CDCl.sub.3): 7.70-7.67 (m, 2H), 7.40-7.38 (m, 2H), 6.58 (bs, 1H), 6.2 (bs, 1H), 5.6 (bs, 1H), 3.86 (s, 2H), 3.63-3.60 (q, 2H), 3.58 (s, 3H), 2.91 (t, J=7.04 Hz; 2H). Mass (M+H): 237.1. IR (cm.sup.1): 3339, 3162, 1661, 1543, 1199, 1120, 690. HPLC purity (%): 95.95 (Max plot), 95.29 (215 nm).

(352) Preparation of Compound (31)

(353) ##STR00195##

(354) Compound (31) was prepared according to the procedure shown in Scheme 23.

(355) ##STR00196##

(356) Preparation of Intermediate BM

(357) ##STR00197##

(358) To a solution of Intermediate BL (5.0 g, 33.29 mmol) and benzoyl peroxide (0.4 g, 1.66 mmol) in CCl.sub.4 (60.0 mL) was added NBS (5.75 g, 33.29 mmol). The reaction mixture was stirred at 70 C. for 4 hours. The reaction mixture was filtered, and the filtrate was concentrated to obtain the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 1% ethyl acetate in petroleum ether as the eluent to afford Intermediate BM (6.8 g, 87%) as pale yellow oil. .sup.1H NMR (CDCl.sub.3): 8.07 (s, 1H), 8.00-7.96 (m, 1H), 7.60-7.58 (m, 1H), 7.43 (t, J=7.67 Hz, 1H), 4.52 (s, 2H), 3.93 (s, 3H).

(359) Preparation of Intermediate BN

(360) ##STR00198##

(361) To a cold solution of Intermediate BM (7.2 g, 31.44 mmol) in DMSO (35.0 mL) at 0 C. was added a sodium cyanide (3.0 g, 62.88 mmol) portionwise over 15 minutes. After the addition was complete, the reaction mixture was allowed to stir at 10 C. for 3 hours. Ice cold water was added to the reaction mixture (50 mL), and the reaction was extracted with ethyl acetate (350 mL), washed with water (225 mL) and brine (25 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 10% ethyl acetate in petroleum ether as the eluent to afford Intermediate BN (2.8 g, 51%) as a pale brown liquid. .sup.1H NMR (CDCl.sub.3): 9.44-9.42 (m, 2H), 8.90-8.78 (m, 2H), 4.63 (s, 3H), 4.49 (s, 2H). Mass (MH): 174.1.

(362) Preparation of Intermediate BO

(363) ##STR00199##

(364) To a cold (0 C.) solution of Intermediate BN (500 mg, 2.85 mmol) in THF (25.0 mL) was added NaBH.sub.4 (216 mg, 5.71 mmol), and the reaction stirred at 80 C. for 15 minutes. Methanol (8.0 mL) was then added to the reaction mixture at 80 C. until the effervescence eased. The reaction was then stirred for approximately 16 hours. The reaction mixture was then quenched with water (10 mL) and concentrated under reduced pressure. The resulting aqueous residue was diluted with water (25 mL) and extracted with ethyl acetate (250 mL). The combined ethyl acetate layers were washed with water (25 mL) and brine solution (25 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 12% ethyl acetate in petroleum ether as the eluent to afford Intermediate BO (410 mg, 95%) as as pale yellow oil. .sup.1H NMR (CDCl.sub.3): 7.40-7.32 (m, 3H), 7.27 (s, 1H), 4.72 (d, J=5.66 Hz, 2H), 3.76 (s, 2H), 1.72 (t, J=5.76 Hz, 1H). Mass (MH): 146.1.

(365) Preparation of Intermediate BP

(366) ##STR00200##

(367) A suspension of Intermediate BO (200 mg, 1.32 mmol) and Raney-Ni (50 mg, wet) in methanolic NH.sub.3 (6.0 mL) was hydrogenated by bubbling H.sub.2 at 10 C. for 3 hours. The reaction mixture was filtered, and the cake was washed with methanol (310 mL). The combined filtrates were concentrated under reduced pressure to afford Intermediate BP (200 mg, crude), as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.30-7.21 (m, 4H), 4.69 (s, 2H), 3.49 (s, 2H), 2.9 (bs, 2H), 4.80 (s, 2H). Mass (M+H): 152.0.

(368) Preparation of Intermediate BQ

(369) ##STR00201##

(370) To a cold (0 C.) solution of Intermediate BP (200 mg, 132 mmol) and triethylamine (0.27 mL, 1.92 mmol) in dichloromethane (10.0 mL) was added slowly methoxyacetyl chloride (0.18 mL, 1.96 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 3 hours. The reaction mixture was diluted with dichloromethane (10 mL), washed with water (220 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 5-10% ethyl acetate in petroleum ether as the eluent to afford Intermediate BQ (250 mg, 64%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.33-7.17 (m, 4H), 6.56 (bs, 1H), 5.18 (s, 2H), 4.08 (s, 2H), 3.87 (s, 2H), 3.58-3.53 (m, 2H), 3.46 (s, 3H), 3.36 (s, 3H), 2.85 (t, J=7.07 Hz; 2H). Mass (M+H): 295.9.

(371) Preparation of Compound (31)

(372) ##STR00202##

(373) A suspension of Intermediate BQ (250 mg, 0.84 mmol) and K.sub.2CO.sub.3 (130 mg, 0.94 mmol) in methanol (3.0 mL) was stirred at 26 C. for 1 hour. The reaction mixture was concentrated under reduced pressure; the residue was then diluted with water (20 mL) and extracted with ethyl acetate (220 mL). The combined ethyl acetate layers were washed with water (10 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (100-200 mesh silica gel) using 20% ethyl acetate in petroleum ether as the eluent to afford Compound (31) (120 mg, 63%) as a pale brown solid. .sup.1H NMR (CDCl.sub.3): 7.32-7.22 (m, 3H), 7.13 (d, J=7.41 Hz; 1H), 6.56 (bs, 1H), 4.67 (s, 2H), 3.85 (s, 2H), 3.58-3.53 (m, 3H), 3.35 (s, 3H), 2.84 (t, J=7.12 Hz; 2H). Mass (M+H): 223.9. IR (cm.sup.1): 3401, 2931, 1658, 1540, 1116, 791. HPLC purity (%): 98.9 (Max plot), 96.92 (254 nm), 97.11 (215 nm).

(374) Synthesis of Compound (32)

(375) ##STR00203##

(376) Compound (32) was synthesized according to the procedure shown in Scheme 24.

(377) ##STR00204##

(378) Preparation of Intermediate BS

(379) ##STR00205##

(380) A suspension of Intermediate BR (1 g, 8.19 mmol) and ammonium acetate (1.38 g, 18.03 mmol) in nitromethane (70.0 mL) was stirred at 90 C. for 1 hour. The reaction mixture was concentrated under reduced pressure, the resulting crude material was dissolved in ethyl acetate (50 mL), washed with water (220 mL) and brine solution (25 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 10% ethyl acetate in petroleum ether as the eluent to afford Intermediate BS (550 mg, 40.6%) as yellow solid. .sup.1H NMR (CDCl.sub.3): 7.95 (d, J=13.68 Hz; 1H), 7.55 (d, J=13.68 Hz; 1H), 7.13 (d, J=7.87 Hz, 1H), 7.01-6.95 (m, 2H), 4.97 (s, 2H). Mass (MH): 164.0.

(381) Preparation of Intermediate BT

(382) ##STR00206##

(383) To a cold (0 C.) solution of Intermediate BS (350 mg, 2.12 mmol) in methanol (3.5 mL) at 0 C. was added portionwise NaBH.sub.4 (100 mg, 2.54 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to stir at 10 C. for 1 hour. The reaction mixture was quenched with water (5 mL) and concentrated under reduced pressure. The resulting aqueous residue was diluted with water (15 mL) and extracted with ethyl acetate (220 mL). The combined ethyl acetate layers were washed with water (10 mL) and brine solution (15 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 10% ethyl acetate in petroleum ether as the eluent to afford Intermediate BT (200 mg, 56.4%) as a pale yellow solid. .sup.1H NMR (CDCl.sub.3): 7.19 (s, J=7.90 Hz; 1H), 6.77-6.72 (m, 2H), 6.68 (s, 1H), 4.59 (t, J=7.32 Hz, 2H), 3.27 (t, J=7.32 Hz). Mass (MH): 166.0.

(384) Preparation of Intermediate BU

(385) ##STR00207##

(386) A suspension of Intermediate BT (150 mg, 0.89 mmol) and 10% Pd/C (25 mg, dry) in MeOH (1.0 mL) was hydrogenated by bubbling H.sub.2 at 10 C. for 3 hours. The reaction mixture was filtered, and the cake was washed with methanol (35 mL). The combined filtrates were concentrated under reduced pressure. The obtained gummy material was dissolved in ethyl acetate (1 mL), treated with EtOAc and HCl (0.5 mL). The reaction stirred for 10 minutes and was then concentrated to afford the HCl salt of Intermediate BU (200 mg, crude) as a pale brown gum. This material was used without further purification in the next step.

(387) Preparation of Intermediate BV

(388) ##STR00208##

(389) To a cold (0 C.) solution of Intermediate BU (200 mg, 1.15 mmol) and triethylamine (0.323 mL, 2.3 mmol) in dichloromethane (3.0 mL) was added slowly methoxyacetyl chloride (0.136 mL, 1.26 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 2 hours. The reaction mixture was diluted with dichloromethane (10 mL), washed with water (220 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (silica gel 100-200 mesh) using 2% MeOH in chloroform as the eluent to afford Intermediate BV (20 mg, 61.7%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.33 (t, J=7.86 Hz; 1H), 7.09 (d, J=7.64 Hz; 1H), 7.00-6.97 (m, 2H), 6.56 (bs, 1H), 4.28 (s, 2H), 3.87 (s, 2H), 3.59-3.54 (m, 5H), 3.35 (s, 3H), 2.85 (t, J=7.04 Hz; 2H). Mass (M+H): 282.0.

(390) Preparation of Compound (32)

(391) ##STR00209##

(392) A suspension of Intermediate BV (200 mg, 0.71 mmol) and K.sub.2CO.sub.3 (108 mg, 0.78 mmol) in methanol (2.0 mL) was stirred at 26 C. for 1 hour. The reaction mixture was concentrated under reduced pressure; the residue was then diluted with water (10 mL) and extracted with ethyl acetate (220 mL). The combined ethyl acetate layers were washed with water (10 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product. This material was purified by column chromatography (100-200 mesh silica gel) using 4% MeOH in chloroform as the eluent to afford Compound (32) (80 mg, 53.7%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.17 (t, J=8.30 Hz; 1H), 6.77 (d, J=7.81 Hz; 1H), 6.71-6.69 (m, 2H), 6.55 (bs, 1H), 4.85 (s, 1H), 3.87 (s, 2H), 3.58-3.53 (m, 3H), 3.35 (s, 3H), 2.79 (t, J=7.08 Hz; 2H). Mass (M+H): 210.0. IR (cm.sup.1): 3392, 2936, 1658, 1542, 1455, 1116, 783. HPLC purity (%): 94.43 (Max plot), 94.16 (215 nm).

(393) Synthesis of Compounds (33), (34), and (35)

(394) ##STR00210##

(395) Compounds (33)-(35) were synthesized according to the procedure shown in Scheme 25.

(396) ##STR00211##

(397) Preparation of Intermediate BX

(398) ##STR00212##

(399) To a cold suspension of NaBH.sub.4 (1.5 mmol) in THF (30 mL) at 0 C., Intermediate BW (1.0 mmol) was added portionwise over 15 minutes. After the addition was complete, the reaction mixture was allowed to stir at room temperature for 2 hours. To the reaction mixture was added slowly a solution of BF.sub.3.O(Et).sub.2 (2.0 mmol) in THF (10 mL) over 3 hours. After the addition was complete, the reaction mixture was stirred at room temperature for 20 hours. The reaction mixture was quenched with 1.5N HCl (10 mL) and MeOH (20 mL) then concentrated. The obtained aqueous residue was basified (pH10) with 1N NaOH solution and extracted with ethyl acetate (330 mL). The combined ethyl acetate layers were washed with brine solution (220 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate BX (Table 17), which was used in the next step without further purification.

(400) TABLE-US-00017 TABLE 17 BX1 Intermediate BW1 (1.5 g, 9.52 mmol) was reacted with NaBH.sub.4 (1.08 g, 28.57 mmol) and BF.sub.3O(Et).sub.2 (4.8 mL, 38.09 mmol) in THF (100 mL) to give Inter- mediate BX1 (1.3 g, crude) as a color- less thick oil. .sup.1H NMR (CDCl.sub.3): 7.66 (t, J = 7.67 Hz; 1H), 7.25 (d, J = 7.87 Hz; 2H), 4.75 (s, 2H), 3.41 (bs, 1H). Mass (M + H): 144.0. BX2 Intermediate BW2 (2.5 g, 15.87 mmol) was reacted with NaBH.sub.4 (0.9 g, 23.80 mmol) and BF.sub.3O(Et).sub.2 (3.9 mL, 31.74 mmol) in THF (100 mL) to give Intermediate BX2 (3.0 g, crude) as a colorless thick oil. .sup.1H NMR (CDCl.sub.3): 8.35 (d, J = 4.87 Hz; 1H), 7.36 (s, 1H), 7.21 (d, J = 5.85 Hz; 2H), 4.75 (d, J = 5.36 Hz; 2H). Mass (M + H): 144.0.

(401) Preparation of Intermediate BY

(402) ##STR00215##

(403) To a cold solution of Intermediate BX (1.0 mmol) in dichloromethane (15 mL) at 0 C. was added slowly thionyl chloride (1.0 mmol) over 15 minutes. After the addition was complete, the reaction mixture was allowed to stir at room temperature for 20 hours. The reaction mixture was cooled, and ice cold water was added (30 mL). The mixture was extracted with dichloromethane (350 mL), washed with water (25 mL) and brine (25 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate BY (Table 18), which was purified by column chromatography (silica gel 100-200 mesh) using 20% ethyl acetate in petroleum ether as the eluent.

(404) TABLE-US-00018 TABLE 18 BY1 Intermediate BX1 (1.3 g, 9.09 mmol) was reacted with thionyl chloride (0.66 mL, 13.98 mmol) in dichloromethane (30 mL) to give Intermediate BY1 (800 mg, 57%) as a pale brown solid. .sup.1H NMR (CDCl.sub.3): 7.69 (t, J = 7.87 Hz; 1H), 7.43 (d, J = 7.46 Hz; 1H), 7.28 (d, J = 7.87 Hz; 1H), 4.63 (s, 2H). Mass (M + H): 161.9. BY2 Intermediate BX2 (2.0 g, 13.98 mmol) was reacted with thionyl chloride (1.0 mL, 13.98 mmol) in dichloromethane (30 mL) to give Intermediate BY2 (1.5 g, 68%) as a pale brown solid. .sup.1H NMR (CDCl.sub.3): 8.39 (d, J = 4.97 Hz; 1H), 7.38 (s, 1H), 7.25 (d, J = 4.56 Hz; 1H), 4.52 (s, 2H). Mass (M + H): 162.0.

(405) Preparation of Intermediate BZ

(406) ##STR00218##

(407) To a cold solution of Intermediate BY (1.0 mmol) in DMSO (10 mL) at 0 C. was added sodium cyanide (2.0 mmol) portionwise over 15 minutes. After the addition was complete, the reaction mixture was allowed to stir at 10 C. for 2 hours. Ice cold water was added to the reaction mixture (30 mL), and the mixture was extracted with ethyl acetate (350 mL), washed with water (25 mL) and brine (25 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate BZ (Table 19), which was purified by column chromatography (silica gel 100-200 mesh) using 10% ethyl acetate in petroleum ether as the eluent.

(408) TABLE-US-00019 TABLE 19 BZ1 Intermediate BY1 (800 mg, 4.93 mmol) was reacted with sodium cyanide (484 mg, 9.87 mmol) in DMSO (8 mL) to give Interme- diate BZ1 (400 mg, 53%) as a pale brown solid. .sup.1H NMR (CDCl.sub.3): 7.73 (t, J = 7.81 Hz; 1H), 7.42 (d, J = 7.61 Hz; 1H), 7.33 (d, J = 8.00 Hz; 1H), 3.93 (s, 2H). Mass (M + H): 152.9.IR (cm.sup.1): 3079.79, 2923.5, 2252.12, 1439.39 and 789.42. BZ2 0 Intermediate BY2 (1.5 g, 9.25 mmol) was reacted with sodium cyanide (0.9 g, 18.51 mmol) in DMSO (15 mL) to give Interme- diate BZ2 (300 mg, 16.6%) as a pale brown solid. .sup.1H NMR (CDCl.sub.3): 8.43 (d, J = 5.07 Hz; 1H), 7.36 (s, 1H), 7.24 (d, J = 5.07 Hz; 1H), 3.78 (s, 2H). Mass (M + H): 153.0. IR (cm.sup.1): 2923.5, 2245.9, 1595.3, 1404.6 and 825.9. BZ3 Intermediate BY3 (1.5 g, 9.25 mmol) was reacted with sodium cyanide (0.9 g, 18.51 mmol) in DMSO (15 mL) to give Interme- diate BZ3 (800 mg, 57%) as a pale brown solid. .sup.1H NMR (CDCl.sub.3): 8.37 (s, 1H), 7.68 (d, J = 8.29 Hz; 1H), 7.39 (s, J = 8.29 Hz; 1H), 7.25 (d, J = 4.56 Hz; 2H), 3.77 (s, 2H). Mass (M + H): 153.0. IR (cm.sup.1): 2923.5, 1439.3, 1108.8 and 789.4.

(409) Preparation of Intermediate CA

(410) ##STR00222##

(411) To a cold solution of Intermediate BZ (1.0 mmol) in THF (30 mL) at 0 C. was BH.sub.3.DMS (9.0 mmol) was added slowly over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 20 hours. The reaction mixture was cooled in ice and then quenched with MeOH (5 mL). The reaction was then refluxed for 1 hour and concentrated. The residue was dissolved in ethyl acetate (50 mL), washed with water (210 mL) and brine solution (15 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate CA (Table 20), which was in the next step without further purification.

(412) TABLE-US-00020 TABLE 20 CA1 Intermediate BZ1 (360 mg, 2.36 mmol) was reacted with BH.sub.3DMS (2.0 mL, 21.24 mmol) in THF (12 mL) to give Intermediate CA1 (250 mg, crude) as a pale brown gum. Mass (M + H): 157.0. CA2 Intermediate BZ2 (350 mg, 2.30 mmol) was reacted with BH.sub.3DMS (1.9 mL, 20.72 mmol) in THF (10 mL) to give Intermediate CA2 (350 mg, crude) as a pale brown gum Mass (M + H): 157.0. CA3 Intermediate BZ3 (1.0 g, 6.57 mmol) was reacted with BH.sub.3DMS (5.6 mL, 59.13 mmol) in THF (30 mL) to give Intermediate CA3 (1 g, crude) as a pale brown gum. Mass (M + H): 157.0.

(413) Preparation of Intermediate CB

(414) ##STR00226##

(415) To a cold (0 C.) solution of Intermediate CA (1.0 mmol) and triethylamine (2.0 mmol) in dichloromethane (20 mL) was added slowly methoxyacetyl chloride (1.1 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 4 hours. The reaction mixture was diluted with dichloromethane (10 mL), washed with water (215 mL) and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate CB (Table 21), which was purified by PREP-TLC plate using 5% MeOH in ethyl acetate as the eluent.

(416) TABLE-US-00021 TABLE 21 CB1 Intermediate CA1 (370 mg, 2.37 mmol) was reacted with methoxy- acetyl chloride (0.24 mL, 2.60 mmol) and Et.sub.3N (0.67 mL, 4.74 mmol) in CH.sub.2Cl.sub.2 (10.0 mL) to give Intermediate CB1 (150 mg 27%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 7.58 (t, J = 7.6 Hz; 1H), 7.20 (d, J = 8.2 Hz; 1H), 7.11 (d, J = 7.4 Hz; 1H), 3.88 (s, 2H), 3.71 (t, J = 6.2 Hz; 2H), 3.40 (s, 3H), 3.0 (t, J = 6.4 Hz; 2H). Mass (M + H): 229.0. CB2 Intermediate CA2 (360 mg, 2.30 mmol) was reacted with methoxyacetyl chloride (0.24 mL, 2.54 mmol) and Et.sub.3N (0.65 mL, 4.61 mmol) in CH.sub.2Cl.sub.2 (10.0 mL) to give Intermediate CB2 (190 mg 35%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 8.31 (t, J = 4.9 Hz; 1H), 7.19 (s, 1H), 7.08 (d, J = 4.56 Hz; 1H), 6.61 (bs, 1H), 3.88 (s, 2H), 3.58 (t, J = 6.6 Hz; 2H), 3.38 (s, 3H), 2.86 (t, J = 7.21 Hz; 2H). Mass (M + H): 229.0. CB3 Intermediate CA3 (1 g, 6.41 mmol) was reacted with methoxyacetyl chloride (0.65 mL, 7.05 mmol) and Et.sub.3N (1.79 mL, 12.8 mmol) in CH.sub.2Cl.sub.2 (20.0 mL) to give Intermediate CB3 (700 mg 50%) as a pale brown gum. .sup.1H NMR (CDCl.sub.3): 8.23 (d,, J = 2.48 Hz; 1H), 7.52 (dd, J = 8.2, 2.48 Hz; 1H), 7.28 (d, J = 7.87 Hz, 1H), 6.61 (bs, 1H), 187 (s, 2H), 3.55 (q, J = 6.6 Hz; 2H), 3.38 (s, 3H), 3.00 (t, J = 6.42 Hz; 2H). Mass (M + H): 229.0.

(417) Preparation of Compounds (33)-(35)

(418) ##STR00230##

(419) To a solution of Intermediate CB (1.0 mmol) in toluene (10 mL) were added sequentially NaOtBu (1.4 mmol), (1) BINAP (0.02 mmol), Pd.sub.2(dba).sub.3 (0.01 mmol), and benzophenone imine (1.2 mmol). The mixture was degassed with argon for 30 minutes and stirred at 80 C. for 6 hours. The reaction mixture was concentrated, and the crude imine was purified by column chromatography (silica gel 100-200 mesh) using 20% MeOH in chloroform as the eluent. The resulting intermediate was dissolved in MeOH (15 mL), and a hydroxylamine solution (50% in water, 1.2 mmol) was added, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated; the resulting aqueous residue was then diluted with water (10 mL) and extracted ethyl acetate (215 mL). The combined ethyl acetate layers were washed with water (10 mL) and brine (15 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product, which was purified using PREP TLC by eluting with 3% MeOH in chloroform. The product amine was treated with EtOAc.HCl and Compounds (33)-(35) were obtained as the corresponding HCl salt (Table 22).

(420) TABLE-US-00022 TABLE 22 (33) Intermediate CB1 (200 mg, 0.87 mmol) was reacted with NaOtBu (117 mg, 1.22 mmol), () BINAP (11 mg, 0.017 mmol), Pd.sub.2(dba).sub.3 (9 mg, 0.008 mmol), benzo- phenone imine (0.18 mL, 1.1 mmol), toluene (4.0 mL), and NH.sub.2OH (0.2 mL) in MeOH (2.0 mL) to give Compound (33)HCl. Yield: 35 mg (20.5%) Pale brown solid. .sup.1H NMR (DMSO-d.sub.6): 13.84 (bs, 1H), 8.00 (t, J = 5.49 Hz; 1H), 7.82- 7.78 (m, 2H), 6.82 (d, J = 8.79 Hz; 1H), 6.65 (d, J = 7.47 Hz; 1H), 3.75 (s, 2H), 3.47-3.41 (m, 4H), 3.25 (s, 3H), 2.85 (t, J = 6.59 Hz; 2H). Mass (M + H): 210.0. IR (cm.sup.1): 3422, 3316, 3145, 1660, 1557, 1118, 790. HPLC purity (%): 98.9 (Max plot), 96.95 (254 nm), 98.91 (215 nm). (34) Intermediate CB2 (200 mg, 0.87 mmol) was reacted with NaOtBu (117 mg, 1.22 mmol), () BINAP (11 mg, 0.017 mmol), Pd.sub.2(dba).sub.3 (9 mg, 0.008 mmol), benzophenone imine (0.18 mL, 1.1 mmol), toluene (4.0 mL), and NH.sub.2OH (0.2 mL) in MeOH (2.0 mL) to give Compound (34)HCl. Yield: 40 mg (18.5%) as a pale brown gum. .sup.1H NMR (DMSO-d.sub.6): 13.52 (bs, 1H), 8.96 (bs, 2H), 7.86 (d, J = 7.04 Hz; 1H), 6.74 (d, J = 4.97 Hz; 1H), 3.76 (s, 2H), 3.42-3.34 (m, 3H), 3.27 (s, 3H), 2.76 (t, J = 6.84 Hz; 2H). Mass (M + H): 210.0. IR (cm.sup.1): 3367, 2926, 1662, 1550, 1114, 815. HPLC purity (%): 94.32 (Max plot), 92.04 (215 nm). (35) Intermediate CB3 (400 mg, 1.75 mmol) was reacted with NaOtBu (236.6 mg, 2.45 mmol), () BINAP (21.8 mg, 0.035 mmol), Pd.sub.2(dba).sub.3 (18.1 mg, 0.017 mmol), benzo- phenone imine (0.35 mL, 2.10 mmol), toluene (8.0 mL), and NH.sub.2OH (0.5 mL) in MeOH (4.0 mL) to give Compound (35)HCl. Yield: 35 mg (16.5%) Pale brown solid. .sup.1H NMR (DMSO-d.sub.6): 13.65 (bs, 1H), 7.90-7.75 (m, 5H), 6.94 (d, J = 9.23 Hz; 1H), 3.75 (s, 2H), 3.38-3.26 (m, 5H), 2.76 (t, J = 6.59 Hz; 2H). Mass (M + H): 210.0. IR (cm.sup.1): 3415, 1668, 1630, 1115, 591. HPLC purity (%): 98.74 (Max plot), 96.57 (254 nm), 98.61 (215 nm).
Synthesis of Compounds (36)-(39)

(421) ##STR00234##

(422) Compounds (36)-(39) were synthesized according to the procedure shown in Scheme 26.

(423) ##STR00235##

(424) Preparation of Intermediate CD

(425) ##STR00236##

(426) To a cold (0 C.) solution of Intermediate CC (1.0 mmol) and triethylamine (1.2 mmol) in dichloromethane (20 mL) was added slowly the requisite sulfonyl chloride (1.2 mmol) over 5 minutes. After the addition was complete, the reaction mixture was allowed to reach room temperature and stirred for 3 hours. The reaction mixture was then diluted with dichloromethane (25 mL), washed with water (225 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude Intermediate CD (Table 23), which was purified by column chromatography (silica gel 100-200 mesh) using 2% MeOH in chloroform as the eluent.

(427) TABLE-US-00023 TABLE 23 CD1 Intermediate CC (250 mg, 1.65 mmol) was reacted with methane- sulfonyl chloride (0.15 mL, 1.92 mmol) and Et.sub.3N (0.28 mL, 1.99 mmol) in CH.sub.2Cl.sub.2 (5.0 mL) to give Intermediate CD1 (350 mg, 92%) as a pale brown oil. .sup.1H NMR (CDCl.sub.3): 7.24 (m, 1H), 6.80-6.75 (m, 3H), 4.22 (bs, 1H), 3.80 (s, 3H), 3.43-3.38 (m, 2H), 2.87-2.84 (m, 5H). Mass (M + H): 230.0. CD2 Intermediate CC (250 mg, 1.65 mmol) was reacted with ethane- sulfonyl chloride (0.18 mL, 1.89 mmol) and Et.sub.3N (0.27 mL, 1.92 mmol) in CH.sub.2Cl.sub.2 (5.0 mL) to give Intermediate CD2 (320 mg, 81%) as a pale brown oil. .sup.1H NMR (CDCl.sub.3): 7.26-7.22 (m, 1H), 6.81-6.75 (m, 3H), 4.05 (bs, 1H), 3.80 (s, 3H), 3.41- 3.36 (m, 2H), 2.97 (q, 2H), 2.85 (t, J = 6.6 Hz; 2H), 1.26 (t, J = 6.6 Hz; 3H). Mass (M H): 242.0. CD3 Intermediate CC (250 mg, 1.65 mmol) was reacted with iso- propylsulfonyl chloride (0.22 mL, 1.96 mmol) and Et.sub.3N (0.28 mL, 1.97 mmol) in CH.sub.2Cl.sub.2 (5.0 mL) to give Intermediate CD3 (340 mg, 82%) as a pale brown oil. .sup.1H NMR (CDCl.sub.3): 7.24 (t, J = 7.5 Hz; 1H), 6.80-6.75 (m, 3H), 3.98 (bs, 1H), 3.80 (s, 3H), 3.39 (q, 2H), 3.12-3.08 (m, 1H), 2.85 (t, J = 6.8 Hz; 2H), 1.32 (s, 3H), 1.30 (s, 3H),. Mass (M H): 242.1. Mass (M + H): 258.0. CD4 0 Intermediate CC (100 mg, 0.66 mmol) was reacted with isobutylsulfonyl chloride (0.11 mL, 0.78 mmol) and triethylamine (0.11 mL, 0.78 mmol) in dichloromethane (5.0 mL) to give Intermediate CD4 (130 mg, 72%) as a pale brown oil. .sup.1H NMR (CDCl.sub.3): 7.24 (t, J = 7.8 Hz; 1H), 6.80- 6.74 (m, 3H), 4.15 (bs, 1H), 3.80 (s, 3H), 3.37 (q, 2H), 2.86- 2.80 (m, 4H), 2.17 (m, 1H), 1.06 (s, 3H), 1.04 (s, 3H). Mass (M + H): 272.0.

(428) Preparation of Compounds (36)-(39)

(429) ##STR00241##

(430) To a cold (70 C.) solution of Intermediate CD (1.0 mmol) in dichloromethane (20 mL) was added slowly BBr.sub.3 (13 mmol). After the addition was complete, the reaction mixture was allowed to reach 0 C. and stirred for 2 hours. The reaction mixture was quenched with ice cold water (15 mL) and extracted with dichloromethane (230 mL). The combined dichloromethane layers were washed with water (210 mL) and brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, and the solvent was removed to afford the crude product (Table 24). This material was then purified by column chromatography (silica gel 100-200 mesh) using 40% ethyl acetate in petroleum ether as the eluent to afford the desired product.

(431) TABLE-US-00024 TABLE 24 (36) Intermediate CD1 (350 mg, 1.52 mmol) was reacted with BBr.sub.3 (0.18 mL, 1.89 mmol) in dichloromethane (6.0 mL) to give Compound (36) (220 mg, 66%) as a pale brown gum..sup.1H NMR (CDCl.sub.3): 7.19 (t, J = 7.80 Hz; 1H), 6.78- 6.69 (m, 3H), 4.81 (s, 1H), 4.19 (bs, 1H), 3.42-3.38 (m, 2H), 2.88-2.81 (m, 5H). Mass (M H): 214.0. IR (cm.sup.1): 3434, 2930, 1597, 1311, 1144, 973, 521. HPLC purity (%): 97.01 (Max plot), 96.81 (215 nm). (37) Intermediate CD2 (320 mg, 1.31 mmol) was reacted with BBr.sub.3 (0.16 mL, 1.68 mmol) in dichloromethane (5.0 mL) to give Compound (37) (260 mg, 86%) as a pale brown oil. .sup.1H NMR (CDCl.sub.3): 7.18 (t, J = 7.71 Hz; 1H), 6.76- 6.71 (m, 3H), 5.43 (s, 1H), 4.28 (t, J = 5.85 Hz; 1H), 3.39-3.34 (m, 2H), 2.98-2.93 (m, 2H), 2.84-2.79 (m, 2H), 1.27 (t, J = 7.32 Hz; 3H),. Mass (M H): 214.0. IR (cm.sup.1): 3402, 3294, 2931, 1589, 1456, 1312, 1137, 869, 697. HPLC purity (%): 97.63 (Max plot), 97.04 (215 nm). (38) Intermediate CD3 (340 mg, 1.32 mmol) was reacted with BBr.sub.3 (0.16 mL, 1.71 mmol) in dichloromethane (5.0 mL) to give Compound (38) (285 mg, 89%) as a pale brown oil. .sup.1H NMR (CDCl.sub.3): 7.19 (t, J = 7.87 Hz; 1H), 6.79-6.70 (m, 3H), 4.86 (s, 1H), 3.95 (bs, 1H), 3.41-3.36 (q, 2H), 3.12-3.09 (m, 1H), 2.83 (t, J = 6.63 Hz; 2H), 1.32 (s, 3H), 1.30 (s, 3H),. Mass (M H): 242.1. IR (cm.sup.1): 3414, 2928, 1589, 1456, 1308, 1132, 885, 783, 696. HPLC purity (%): 94.44 (Max plot), 93.92 (215 nm). (39) Intermediate CD4 (130 mg, 0.47 mmol) was reacted with BBr.sub.3 (0.06 mL, 0.61 mmol) in dichloromethane (5.0 mL) to give Compound (39) (110 mg, 89%) as a pale brown oil. .sup.1H NMR (CDCl.sub.3): 7.19 (t, 1H), 6.69 (m, 3H), 4.83 (s, 1H), 4.05 (t, 1H), 3.38 (q, 2H), 2.84- 2.81 (m, 4H), 2.2 (m, 1H), 1.06 (s, 3H), 1.05 (s, 3H),. Mass (M H): 256.1. IR (cm.sup.1); 3413, 2963, 1589, 1457, 1308, 1139, 869, 784, 696. HPLC purity (%): 97.14 (Max plot), 97.28 (215 nm).
Screening Conditions for Identifying SPR Inhibition

(432) The compounds described herein were screened for activity as inhibitors of Sepiapterin Reductase (SPR).

(433) Protein Production

(434) His-tagged recombinant human SPR (GenBank accession number. NM_003124) was cloned as synthetic gene and expressed in E. coli Rosetta 2 strain. Bacteria were grown at 37 C., and expression of SPR protein was induced for 4 hours. After cell lysis, the His-tagged SPR protein was affinity purified with a TALON column (purity of the isolated SPR protein is >95%). In an initial quality control, the enzymatic activity of recombinant SPR was confirmed with a chromogenic assay (read-out OD at 420 nm).

(435) Primary Screen

(436) To screen for SPR inhibition, a biochemical assay based on LC/MS (and chromogenic) read-out has been developed. The LC/MS assay monitors the product formation (L-biopterin) and the chromogenic assay measures OD at 420 nm.

(437) N-methoxyacetyl serotonin was used as a reference compound (positive control). The IC.sub.50 measured using the screening conditions was 20-40 nM, which agrees with the literature (Smith et al., Journal of Biological Chemistry, 297:5601, 1992).

(438) The exemplary assay protocol uses the following conditions: SPR (6 nM); L-Sepiapterin (50 M); NADPH (100 M); Na-Phosphate buffer, pH 6.5 (100 mM); 82 L assay volume; 60 minutes incubation with compounds (0.5% final concentration in DMSO) at 37 C. in Greiner clear 384 well plates.

(439) The following experimental procedure was applied: (1) Add 2 L compound (inhibitor) dilutions (20% DMSO) in Greiner clear 384 well plates. (2) Add 40 L enzyme/assay buffer. (3) Start: 40 L substrate solution/assay buffer. (4) Final: 82 L assay solution. (5) Incubation: Safire 1 hour at 37 C. and measuring after 1 hour using OD at 420 nm (chromogenic read-out). (6) Transfer 50 L to a 384 Matrix flat bottom (clear) for LC/MS measurement. (7) Stop: add 5 L 1M HCl and 10 L of 0.1 M I.sub.2/NaI solution. (8) Incubation: 45 minutes at 37 C. (9) Neutralization: 10 L 0.1 M ascorbic acid and 5 L 1 N NaOH. (10) LC/MS measurement.

(440) If desired, the compounds can be been further screened using an 8 point dilution series to validate the results. For example, the compounds of Table 1 were screened using this additional method in triplicate. These tests were performed at the following concentrations:

(441) Most potent: 0.2-0.7-2.1-6.2-18.5-55.6-166.7-500 nM

(442) Medium potent: 0.002-0.007-0.02-00.6-0.02-0.7-1.7-5 M

(443) Less potent: 0.02-0.07-0.2-0.6-1.9-5.6-16.7-50 M

(444) A robust performance of the SPR assay was achieved throughout the screen, resulting in a mean Z-value of 0.93 (chromogenic) and 0.82 (LC/MS), and the inhibitors showed the expected response in the LC/MS (chromogenic) assay.

(445) Screening of the compounds has been performed at three concentrations (20 nM, 200 nM, and 2000 nM) in singletons (0.5% final concentration in DMSO). Z was 0.91 and 0.81 for the chromogenic and the LC/MS measurements, respectively. The screens showed that compounds of Formulas (I) and (II) can inhibit SPR, even at the lower concentrations. For example, at the 20 nM concentration, up to 77% inhibition of enzyme activity was observed. Exemplary IC.sub.50 values are presented in Table 25.

(446) TABLE-US-00025 TABLE 25 IC.sub.50 chromo- IC.sub.50 genic LC/MS No. Structure [M] [M] control N-Methoxyacetyl-Serotonin 0.042 0.043 (1) 0.40 0.33 (2) 0.36 0.34 (3) 2.8 2.2 (16) 3.4 2.8 (17) 0 4.5 3.2 (18) 1.1 1.0 (21) 0.084 0.086 (22) 0.011 0.019 (23) 0.011 0.012 (24) 0.31 0.31 (32) 0.064 0.069 (12) 2.1 1.6

Other Embodiments

(447) While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure that come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth.

(448) All references, patents, patent application publications, and patent applications cited herein are hereby incorporated by reference to the same extent as if each of these references, patents, patent application publications, and patent applications were separately incorporated by reference herein.