Title of Invention

"COMPOUNDS USEFUL FOR INHIBITING MACROPHAGE MIGRATION INHIBITORY FACTOR"

Abstract The present invention provides a compound having Formula I or II: wherein B, R, X, Ar, and Y are defined herein, pharmaceutically acceptable salts thereof and pharmaceutically acceptable prodrugs thereof. The present invention also provides methods of making and using the compound.
Full Text FIELD OF THE INVENTION
COMPOUNDS, COMPOSITIONS, PROCESSES OF MAKING, AND
METHODS OF USE RELATED TOINHIBITING M ACROPHAGE MIGRAT1ONINHIBITORY FACTOR
This application.claims priority to U.S. Provisional Application Serial No. 60/556,440, filed March 26,2004, the entire contents of which are hereby incorporated by reference.
BACKGROUND OF THE INVENTION FieM of the laveatio»
The present invention relates to isoxazoline and related compounds, to
intermedtates and meflmds for their prqpMratinn in cnmpnsiiining rraf|tqipjţ) andtotheiruse.
Macropbage migration inhibitory factor (MEF) is one of the earliest described cytokines, and is an immunoregulatory protein with a wide variety of cellular and biologica! activities (forreviews see: Swope. et al.. Rev. Physiol. Biochem. Pharmacol. 139,1-32 (1999); Metz. et al.. Adv. Immunol. 66,197-223 (1997); and Bucala. FASEB J. 14,1607-1613 (1996)). Originally, MIF was found to be secreted by activated h/mphoid cells, to inhibit the random migration of macrophages, and to be associated with delayed-type hypersensitivity reactions (Georee. et al.. Proc. Soc. Exp. Biol. Med., 111, 514-521 (1962); Weiser.etal.. J. Immunol. 126,1958-1962 (198n: Bloom. et al.. Science, 153:80-82 (1966); Pavid. Proc. Natl. Acad. Sci. USA, 56, 72-77 (1966). MIF was also shown to

enhance macrophage adherence, phagocytosis and tumoricidal activity fNatbanetai.. J. Exp. Med., 137,275-288 (1973); Nafran, J. Exp. Med., 133,1356-1376 (1971); Churchill. et al. J. Immunol, 115,781-785 (1975)). The availability of recombinant MOP nas allowed for confinnation of these biologica! activities, and for the identification of additional activities.
Recombinant human MIF was originally cloned firom a human T cell library (Weiser. et al.. Proc. Natl. Acad. Sci. USA, 86,7522-7526 (1989)), and was shown to activate blood-derived macrophages to kill intracellular parasites and tumor cells in vitro, to stimulate ÎL-1P and TNFa expression, and to induce nitric oxide synthcsis (Weiser. et al.. J. ImmunoL, 147,2006-2011 (1991); PozzL et al.. Cellular Immunol., 145,372-379 (1992); Weiser. et al.. Proc. Natl Acad. Sci. USA, 89,8049-8052 (1992); ftmff»-** J- Immunol, 150,1908-1912 (1993)). Whtle the conchBÎons ayailabfe from severa! of these earty reports are confounded by the presence of a btoactive ontogenie contaminant in tite recombinant MIF preparatioro used, the potent pro-infiammatory activities of MIF have been established in other studies mat do not suffer firom this complicating factor (reviewed in Bucala. The FASEB, Journal 10,1607-1613 (1996)).
More recent MIF studies have capitalized on the production of recombinant MIF in purified form as well as the development of MIF-specifîc polyclonal and monoclonal antibodies to establish the biological role of MIF in a variety of normal homeostatic and pathophysiological settings (reviewed in Rice. et al.. Annual Reports in Medicinal Chemistry, 33,243-252 (1998)). Among the most important insights of these later reports has been the recognition that MIF not only is a cytokine product of the immune system, but also is a hormone-like product of the
endocrine system, particularly the pitiritary gland. This work has underscored the potent activity of MBP as a counter-regulator of the anti-inflammatory effects of the glucocorticoids (both those endogenousfy released and those therapeutically administered), with the effect mat the normal activities of glucocorticoids to limit and suppress the severity of inflammatory responses are inhibited by MIF. The endogenous MIF response is thus seen as a căuşe or an exacerbative factor in a variety of inflammatory diseases and conditions (reviewed in Donnelly. et al.. Molecular Medicine Today, 3, pp.502-507 (1997)).
MDF is now known to nave severa! biological functions beyond its well-known association with delayed-type hypersensitiviry reactions. For example, as mentioned above, MB? released by macrophages and T cells acts as a prhritary mediator in response to physiological coacentrations of glucocorticoids (Bucj|g, FASEB J., 14,1607-1613 (1996». This Jeads to an overridmg effect of glucocord'coid im»»Mno-«in»ea"vg •erivfty thrrmgh ahraţjong in TNF- o, ILIB, IL-6, and IL-8 levels. Addrtional biological activities of MDF include the regulation of stimulated T cells (pahCT.yţflţ, Proc. Natl. Acad. Sci. USA, 93,7849-7854 (1996)), the control of IgE synthesis (Mifrrn»WTet fll„ Proc. Natl. Acad. Sci. USA, 90,10056-10060 (1993)), the funcţional inactivation of the p53 tumor suppressor protein (Hudson. et al.. J. Exp. Med, 190,1375-1382 (1999)), the regulation of glucose and carbohydrate metabolism (Sakaue. et al.. Mol. Med., 5, 361-371 (1999)), and the attenuation of tumor cell growth and tumor angiogenesis (Chesnev. et al.. Mol. Med., 5,181-191 (1999); Şhhniqai, et şln Biochem. Biophys. Res. Commun., 264, 751-758 (1999)).
Interleukin-1 (DL-1) and Tumor Necrosis Factor (TNF) are biological
substances produced by a variety of cells, such as monocytes or macrophages. ÎL-1 has been demonstrated to mediate a variety of biological activities thought to be important in immunoregulation and other physiological conditions such as inflammation. The myriad of known biological activities of ÎL-1 include the activation of T helper cells, induction of fever, stimulation of prostaglandin or collagenase production, neutrophil chemotaxis, induction of acute phase proteins and the suppression of plasma iron levels.
There are many disease states in which excessive or unregulated ÎL-1 production is implicated in exacerbating and/or causing the disease. These include rheumatoid arthritis, osteoarthritis, endotoxemia and/or toxic shock syndrome, other acute or chronic inflammatory disease states such as the inflammatory reaction induced by endotoxin or inflammatory bowel disease, tuberculosis, atherosclerosis, diabetes, muscle degeneration, cachexia, psoriatic arthritis, Reiter's syndrome, rheumatoid arthritis, gout, traumatic arthritis, rubella arthritis, and acute synovitis.
Excessive or unregulated TNF production has been implicated in mediating or exacerbating a number of diseases including rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and other arthritic conditions; sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, adult respiratory distress syndrome, cerebral malaria, chronic pulmonary inflammatory disease, silicosis, pulmonary sarcoidosis, bone resorption diseases, reperfusion injury, graft vs. host reaction, allograft rejections, fever and myalgias due to infection, such as influenza, cachexia secondary to infection or malignancy, cachexia secondary to acquired immune deficiency syndrome (AIDS), AIDS, ARC
(AIDS related complex), keloid informaticii, scar tissue formation, Crohn's disease, ulcerative colitis, or pyrosis.
Interleukin-8 (IL-8) is a chemotactic factor produced by several cell types including mononuclear cells, fibroblasts, endothelial cells, and keratinocytes. Its production from endothelial cells is induced by ÎL-1, TNF, or lipopolysaccharide (LPS). IL-8 stimulates a number of functions in vitro. It has been shown to have chemoattractant properties for neutrophils, T-lymphocytes, and basophils. In addition it induces histamine release from basophils from both normal and atopic individuala as well lysosomal enzyme release and respiratory burst from neutrophils. IL-8 has also been shown to increase the surface expression of Mac-1 (CD 11 b/CD 18) on neutrophils without de novo protein synthesis, this may contribuie to increased adhesion of the neutrophils to vascular endothelial cells. Many diseases are characterized by massive neutrophil infiltration. Conditions associated with an increase in IL-8 production (which is responsible for chemotaxis of neutrophils into the inflammatory site) would benefit by compounds which are suppressive of IL-8 production.
ÎL-1 and TNF affect a wide variety of cells and tissues and these cytokines as well as other leukocyte derived cytokines are important and criticai inflammatory mediators of a wide variety of disease states and conditions. The inhibition of these cytokines is of benefit in controlling, reducing and alleviating many of these disease states.
The three-dimensional crystal structure of human MIF reveals that the protein exists as a homotrimer (Lolis. et al.. Proc. Ass. Am. Phys., 108, 415-419 (1996) and is structurally related to 4-oxalocrotonate tautomerase, 5-
carboxymethyi-2-hydroxymuconate, chorismate mutase, and to D-dopachrome tautomerase (Swope. et al.. EMBO J., 17, 3534-3541 (1998); Sumoto, et al.. Biochemistry, 38, 3268-3279 (1999). Recently, the crystal structurc has been reported for the complex formed between human MIF and p-hydroxyphenylpyruvic acid flLubetskv. et al.. Biochemistry, 38, 7346-7354 (1999). It was found that the substrate binds to a hydrophobic cavtty at the amino terminus and interacts with Pro- 1, Lys-32, and De-64 in one of the subunits, and with Tyr-95 and Asn-97 in an adjacent subunit Similar interactions between murine MIF and (E)-2-fluoro-p-hydroxycinnamate have been reported (Taylor. et al.. Biochemistry, 38, 7444-7452 (1999)). Solution studies using NMR provide further evidence of the interaction between p-hydroxyphenylpyiuvic acid «nd Pro- 1 in the amino-terminal hydrophobic cavity (Swope. etaL EMBO J., 17, 3534-3541 (1998)).
Mutation studies provide convincing evidence that Pro- 1 îs invorved in the catarytic function of MIF. Defetion of Pro- 1 or replacement of Pro- 1 with Ser
yţ ft|„ Biochemistry, 36,15356-15362 (1997)), Gry (Swope. et al.. EMBO
J., 17, 3534-3541 (1998)), or Phe MermffnnwsKJ-Vpsatka. gţ Biochemistry, 38, 12841-12849 (1999)), and additkm of an N-terminal peptide tag to Pro-1 (Bendrat et al.. Biochemistry, 36, 15356-15362 (1997)) abrogated the catalytic activity of MIF in assays using L-dopachrome methyl ester and p-hydroxyphenylpyruvic acid. A similar loss in acnvity was found by inserting Ala between Pro-1 and Met-2 (Lubetskv et al.. Biochemistry, 38,7346-7354 (1999). The Pro to Ser MIF mutant showed glucocorticoid counter-regulatory activity (Bendrat et al.. Biochemistry, 36,15356-15362 (1997)) and was fully capable, as was the Pro to Phe mutant, of inhibiting monocyte cbemotaxis (Hermanowski-Vosatka et aln
Biochemistry, 38, 12841-12849 (1999). in contrast, Ihe Pro to Gly MIF mutant was greatly impaired in its abilhy to stimulate superoxide generation in acti vated neutrophils (Swopeetal.. EMBO J., 17, 3534-3541 (1998).
MIF bas been characterized as an anterior pituitary-derived hormone that potentiates lethal endotoxemia (Bucala. Immunol. Lett., 1994, 43, 23-26; Bucala. Circ. Shock, 1994, 44, 35-39), a factor which can override glucocorticoid-mediated suppression of inflammatory and immune responses (Calandra and Bucala. Crit. Rev. Immunol., 1997, 17, 77-88; Calandra and Bucala. J. Inflamm., 1995, 47, 39-51), and as an activator of T-cells after mitogenic or antigenic stimuli (Bacher et ai, Proc. Natt. Acad. Sci. U. S. A 19%, 93, 7849-7854).
This cytokine bas been shown to have multiple roles whhin mc confines of regulating the immune rcsponse as wefl as betng associated with cdl growth and dJffercntMrtkTnduringwouDdîepakaDdca»cmogenesis. Expression bas been shown to be devrted in prostate «denocarcinomas (Arcuri et al.. Prostate, 1 999, 39, 159-165; Mever-Sieyler yrf ffmfrm Urotogy, 1996, 48, 448-452), colon carcinomas of the mouse (TftkftlNIlfî ti Mn1 Med., 1998, 4, 707-714), lipopolysacharide-induced HL60 cells (a leukemia cell li
Biochem. Mol. Biol. Int, 1996, 40, 861-869), and upon treatment with ultraviolet radiation (fflimiTf»',, J. Invest Dermatol., 1999, 1 12, 210-215). The pharmacological modulation of MIF activity and/or expression may therefore be an appropriate point of therapeutic intervention in pathological conditions.
The protein has been detected in the synovia of patients with rheumatoid arthritis (Onodera et al.. Cytokine, 1999, 11, 163-167) and its expression at sites of inflammation and from macrophages suggests a role for the mediator in regulating
the function of macrophages in host defense (Calandra et al.. J. Exp. Med., 1994, 179,1895-1902). Activity of MIF has also been found to correlate well with delayed hypersensitivity and cellular immunity in humans flBernhagenetal.. J. Exp. Med., 1996, 183,277-282; David. Proc. Natl. Acad. Sci. U. S. A., 1966, 56, 72-77). The protein has also been implicated in neural function and development in rodents rfiacher et al.. Mol. Med., 1998,4,217-230; Matsunaga et al.. J. Biol. Chem., 1999,274, 3268-3271 :Nishioet al.. Biochim. Biophys. Acta., 1999,1453, 74-82;
i
Suzuki et al.. BrainRes., 1999, 816,457-462).
There is a need in the art to discover and develop small organic molecules that function as MBF inhibitors (e.g., antagonists) and further possess the benefits of small organic molecule therapeutics versus larger, polymeric protein (e.g., antibody) and nucleic acid-based (e.g., anti-sense) therapeutic agents. The therapeutic potenţial of low molecular weight MIF inhibitors is substanţial, given the activities of anti-MIF antibodies in models of endotoxin- and exotoxin-induced toxic shock (Bernhagen et al.. Nature, 365,756-759 (1993); Kobavashi et al.. Hepatology, 29,1752-1759 (1999); Calandra et al.. Proc. Natl. Acad. Sci. USA., 95, 11383-11388 (1998); and Makita et al.. Am. J. Respir. Crit. Care Med. 158, 573-579 (1998), T-cell activation rBacher et al.. Proc. Natl. Acad. Sci. USA., 93, 7849-7854 (1996), autoimmune diseases (e.g., graft versus host disease, insulin-dependent diabetes, and various forms of lupus) including rheumatoid arthritis (Kitaichi. et al.. Curr. Eye Res., 20,109-114 (2000); Leech. et al.. Arthritis Rheum., 42, 1601-1608 (1999), wound healing (Abe. et al.. Biochim. Biophys. Acta, 1500,1-9 (2000), and angiogenesis (Shimizum. et al. Biochem. Biophys. Res. Commun., 264, 751-758 (1999). Low molecular weight anti-MIF drugs
exhibiting such activities may offer clinical advantages over neutralizing antibodies and nucleic acid-based agents because they may be orally active or generally more easily administered, nave better bioavailabilities, nave improved biodistributions, and are normally much less expensive to produce. Related Art
United States Patent No. 4,933,464 to Markofsky discloses a process for forming 3-phenylisoxazolines and 3-phenylisoxazoles and related producte.
United States Patent No. 6,114,367 to Cohan et al. discloses isoxazoline compounds which are inhibitors of tumor necrosis factor (TNF). The isoxazoline compounds are said to be useful for inhibiting TNF in a mammal in need thereof and in the treatment or alleviation of inflammatory conditions or disease. Also disclosed are pharmaceutical compositions comprising such compounds.
Curuzu et al.. Collect. Czech. Chem. Commun., 56: 2494-2499 (1991) discloses 3-substituted phenyl-4,5-dihydroisoxazoleneacetic acids, including 3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazolineacetic acid and 3-(4-methoxyphenyl)-4,5-dihydro-5-isoxazolineacetic acid, and shows that the first of these two compounds is devoid of anti-inflammatory activity, while the second is dramatically reduced in such activity compared to the parent compound that was unsubstituted hi the para position of the phenyl ring, in a carageenin-induced edema assay in the rât paw.
Witvak et al. J. Med. Chem., 40: 50-60 (1997) discloses isoxazoline antagonists of the glycoprotein Hb/IIIa receptor.
Kleimnan. et al.. "Striking effect of hydroxamic acid substitution on the phosphodiesterase type 4 (PDE4) and TNF alpha inhibitory activity of two series of
rolipram analogues: implications for a new active site model of PDE4". J. Med.
Chem. 41(3): 266-270 (1998), discloses inter alia the following compounds:
[33-cyclopeirtyloxy-4TOethoxy-p
and the methyl ester thereof, as well as [3-(3-cyclopentyloxy-4-methoxy-phenyl)-
4,5-iirrydro-isoxazol-5-yl]-M-hydroxy-acetamide.
United States Patent No. 6,492,428, to Al-Abed et al. issued December l O, 2003, and discloses quinone-related compounds having MIF inhibitor activity.
United States Patent No. 6,599,938, to Al-Abed et al. issued July 29,2003, and discloses amino acid/benzaldehyde Schiffbase compounds having MIF inhibitor activity.
United States Patent No. * g*.t~ P.t~* M»
7,2003, and discloses compounds in phannaceutical compositions.
United States Patent Appm. Pub. No. 2003/0008908 to Al-Abed published on January 9,2003 and discloses compounds in phannaceutical compositions.
Any disclosure cited herein is incorporated by reference in its entirety for all purposes.
SUMMARY OF THE INVENTION
One embodiment of the present invention provides a compound having Formula I or H:

(Fiugre Removed)
wherein B is oxygen or sulphur; and each R is indcpendently defined as follows:
wherein in Formula I and Formula H, at kast oue R is not hydrogen;
(Figure Removed)
whendneachR'isindqpendentlybydrogcmana gro cycloalkyl group, a halo group, apeifluoroalk group, a periruoroalkoxy group, an alkenyl group, an alkynyl group, ahydroacy grcraf anoxo group, a mercapto group, an alkyltfak) group, an alkoxy group, an aryi group, a hcteraryl group, an aryloxy group, a heteroaryloxy group, an aralkyl group, a heteroaralkyl group, an aralkoxy group, a heteroaralkoxy group, an HO—(C =O)— group, an amino group, an alkylamino group, a dialkylamino group, a carbamoyl group, an alkylcarbonyl group, an alkoxycarbonyl group, an alkylaminocarbonyl group, a dialkylamino carbonyl group, an arylcarbonyl group, an aryloxycarbonyl group, an alkylsulfonyl group, or an arylsulfonyl group;
each R2 is independentty an alkyl group, a cycloalkyl group, a halo group, a perfluoroalkyl group, a perfluoroalkoxy group, an alkenyl group, an alkynyl group, a hydroxy group, an oxo group, a mercapto group, an alkylthio group, an alkoxy group, an aryl group, a heteroaryl group, an aryloxy group, a heteroaryloxy group, an aralkyl group, a heteroaralkyl group, an aralkoxy group, a heteroaralkoxy group, an HO—(C =O)— group, an amino group, an alkylamino group, a dialkylamino group, a carbamoyl group, an alkylcarbonyl group, an alkoxycarfoonyl group, an alkylaminocarbonyl group, a dialkylamino carbonyl group, an arylcarbonyl group, an aryloxycarbonyl group, an alkylsulfonyl group, or an arylsulfonyl group each m is independentry zero or an integer from one to twenty; and each X is independentry carbon or nitrogen, wherein when any X is carbon, then each Y is defined independentry as follows:

(Figure Removed)
whereineachZismdependentryhydrogen,raalkyl
group, a halo group, aperfluoroalkyl group, aperfluoroalkoxy group, an alkenyl group, an alkynyl group, a hydroxy group, an oxo group, a mercapto group, an alkylthio group, an alkoxy group, an aryl group, a heteroaryl group, an aryloxy group, a heteroaryloxy group, an aralkyl group, a heteroaralkyl group, an aralkoxy group, a heteroaralkoxy group, an HO—(C ==O)— group, an amino group, an alkylamino group, a dialkylamino group, a carbamoyl group, an alkylcarbonyl group, an alkoxycarbonyl group, an alkylaminocarbonyl group, a dialkylamino carbonyl group, an arylcarbonyl group, an aryloxycarbonyl group, an alkylsulfonyl group, or an arylsulfonyl group; and
each n is independentry zero or an integer from one to four;
pharmaceutically acceptable salts thereof and pharmaceutically acceptable
prodrugs thereol.
One embodiment of the present invention provides a method, which inchjdes inhibiting the production of at least one cytokine selected from the group mcludmg MBF, IL-1, IL-2, IL-6, Ib-8, IPN-y, TNF, and a combination thereof in a mammalian subject in need thereof by administering an inhibiting-effective amount of the above compound to the subject
Another embodiment of the present invention provides a method, which includes inhibiting an ERK/MAP pathway in a mammalian subject in need thereof by administering an inhibiting-effective amount of the above compound to the subject
DESCRIPT1ON OF
Various othcr objects, features, and attendant advantages of the present invention will be more fUly appreaated as the same bcxxwaes better understood from the followmg detaikd description when coDfflderedinconnectionwiththe accompanying drawings in which Hke reference characters designate like or corresponding parts throughout the severa! views and wherein:
+
Figure IA shows one synthetic scheme for synthesizing Phenyl Series A Compounds according to one embodiment of the invention;
Figure l B shows one synthetic scheme for synthesizing Phenyl Series B Compounds according to one embodiment of the invention;
Figure 2A shows one synthetic scheme for synthesizing Propyl Series A Compounds according to one embodiment of the invention;
Figure 2B shows one synthetic scheme for synthesizing Propyl Series B Compounds according to one embodiment of the invention;
Figure 3A shows one synthetic scheme for synthesizing Butyl Series A Compounds according to one embodiment of Ine invention;
Figure 3B shows one synthetic scheme for synthesizing Butyl Series B Compounds according to one embodiment of the invention; and
Figure 4 shows one synthetic scheme for synthesizing Furyl Series Compounds according to one embodiment of the invention.
TifTAfILfP DESCRIPTION OF THE EMBODIMENTS
A more complete appreciation of the invention and many of the attendant advantages thereof will be readity obtained as the same becomes better understood by reference to the following detailed descriptkm when considered with the accompanving drawings.
The present inventam relates to isoxazoline and related compounds, to mtennediatesandmethodsibrtheîrpKp«r and to their use. M"** pffMIHy, compositions containmg fhe subject compounds, and medicinal uses of the subject compounds and compositions. Even more particularly, the present invention may be suitably used for the prevention and treatment of various condiţiona in humans. One aspect of the present invention provides for a genus of isoxazoline and isoxazoline- related compounds, pharmaceutical compositions and related methods of making and their use in treatments and diagnostics. The compounds have macrophage migration inhibitory factor (MIF) antagonist activity, and related activities with other cytokines affected by MIF activity. The compounds act as inhibitors of MIF, and also modulating omer cytokines affected by MIF activity including ÎL- 1, IL-2, IL-6, IL-8, IPN-y and TNF. The compounds and
compositions arc useful for treating a variety of diseases involving any disease state in a human, or other mammal, which is exacerbated by or caused by excessive or unregulated MB-, IL-1, IL-2, EL-6, EL-8, IFN-y and TNF production by such mammal's cells, such as, but not limited to, monocytes and/or macrophages, or any disease state that is modulated by inhibiting the ERK/MAP pathway.
hi the following chemical formulae, the use of the superscript on a substituent is to identify a substituent name (e.g.," R2" is used to indicate an R2-named substituent), while the use of a subscript is used to enumerate the number of times a substituent occurs at that molecular site (e.g.," R,, ** or " (R), " both are used to indicate two substituents simpry named as "R").
The present invention refattes to a compound of generai Formula I or n

wherein B is either oxygen or sulphur and each "R" is independently defined:
(Figure Removed)

with the requirement that each 11" cannot only occur as hydrogen on either Formula I or II (i.e., at least one R on either Formula I or n is an "R" substituent other than hydrogen), and any B is independently either oxygen or sulphur; any R1 is independently hydrogen, (Q-Calkyl or some other suitable substituent, any R2 is an amine, an alkoxy or some other suitable substituent; and "m" is independently either zero or an integer from one to twenty;
each X is independently ehher carbon or nitrogen; and when any X is carbon, then Y is the substituent defined independently for each X as


(Figure Removed)
each Z is independently ehher hydrogen, hydroxyl, halogen, or some other siritable substituent; and
"n" is independently ehher zero or an integer from one to four; and pharmaceun'cally acceptable sahs and prodrugs tnereof.
In one embodiment, for compounds having Formulas I and n hereinabove and below,when the ring "X" is nitrogen instead of carbon, then that X nitrogen does not bear a Y. Forexample,mtiibembodinMtfa£niimbcofYgroup5may cocrespond to the number of X carbons, Le a number of l, 2,3 or 4.
In one embodiment, tbe present invention exchides compounds within general Formula I and having a chemical structure &Hing wfthin Formula IA:

(Figure Removed)
wherein
each Y' is independentry a hydrogen or (C,-C6)alkyl;
each Y2 is independently a Y1, hydroxyl, halo, — N3, — CN, — SH, or
ResMs independentry a Y1, halo, — N3, — CN, — OY1,— NCY1,— SH, ==O, =CH2, or A, and each A is independentry either phenyl or an aromatic ring substituted with one or more independent Y2 substituents; Resb is defined as follows:
(Figure Removed)
'isindeiwndentlyaY'.A,—(Oij)-A,—(Yor —NY'Y5, whh each Y5being a saturated or unsjrturated, stndghtor branched (C,-C1E)alkyl; and
Y4 is independently a Y1, —OY1, —OY3, —Z(ZY1) —TY'Y5, or A.
The present invention also relates to the pharmaceutically acceptable acid addition salts of the compounds of general Formula I or n.
The compounds of the Formula I or n which are basic in nature are capable , of forming a wide variety of different salts with various inorganic and organic acids, Aithough such salts must be pharmaceutically acceptable for administration to animals, it is oflen desirable in practice to initially isolate a compound of the Formula I or H from the reaction mixture as a pharmaceutically unacceptable salt and then simply convert the latter back to the free base compound by treatment with an alkaline reagent, and subsequently convert the free base to a
phannaceutically acceptabJe acid addition salt The acid addition saits of tbe base compounds of this invention ara readily prepared by treating the base compound with a substantially equivalent amount of tbe chosen mineral or organic acid in an aqueous solvent medium or in a siritable organic solvent such as methanol or ethanol. Upon caretul evaporation of the solvent» tfae desired solid salt is obtained. The acids which are used to prepare the pharmaceutically acceptable acid addition salts of Ihe aforementioned base compounds of this invention include those which form non-toxic acid addition salts, i.e., sahs containing phannacologically acceptable anions, such as the chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, cifrate, acid cifrate, tartrate, bitartrate, succinate, maleate, fumarate, glutamatc, L-lactate, L-tarfraie, tosylate, mesylate, gtuconate, saccharate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-tohienesuUbnate and pamoate (i-e-» l,rHnethylene-bis-(2-hydroxy-3-naphthoate)>aUts.
The invention also relates to base addition sahs of the compound. The chemical bases that may be used as reagents to prepare pharmaceutically acceptable base salts of those compounds of general Formula I or n that are acidic in nature are those that form non-toxic base salts with such compounds. Those compounds of the Formula I or n which are also acidic in nature, e.g., where substituent R, R1, R2, or R3 includes a -COOH or tetrazole moiety, are capable of forming base salts with various phannacologically acceptable cations. Examples of such salts include the alkali metal or alkaline-earth metal salts and particularly, the sodium and potassium salts. These sahs are all prepared by convenţional techniques. The chemical bases which are used as reagents to prepare the pharmaceutically
acceptable base sahs of this invention include those which fonn non-toxic base safts wrth the herein described acîdic compounds of Formula I or O. These salts caneasilybepreparedbytreatiiigtherorres
aqueous solution containing the desired pharmacologically acceptable cations, and then evaporating the resuWng solution to dryness, preferably under reduced pressure. Altematively, they may also be prepared by mixing lower alkanolic solutions of the acidic compounds and the desired alkali metal alkoxide together, and then evaporating the resulting solution to dryness in the same manner as before. hi either case, stoichiometric quantities of reagents are preferably employed in order to ensure completeness of reaction and ţnffinţtmffi» product yieids. Such non-toxic base săra include» but ne not limited to those derived from such pharmacologicaUy acceptable cations snch as alkah' metal cations (e.g., potassium and sodium) and «IMine evm metal cations (e.g., calcium and magnesium), ammonium or water-sohiWe amine addition sahs such as N-
iiţpEX and the 10W6T *nV«t>olaTrţyrţniniiim and Other base
sahs of pharmaceutically acceptable organic amines.
The compounds and prodrugs of the present invention can exist in several tautomeric fonns, and geometric isomers and mixtures thereof. AII such tautomeric forms are included whhin the scope of the present invention. Tautomers exist as mixtures of tautomers in solution. hi solid fonn, usually one tautomer predominates. Even though one tautomer may be described, the present invention includes all tautomers of the present compounds.
The present invention also includes atropisomers of the present invention. Atropisomers refer to compounds of the invention that can be separated into
rotationally restricted isomers. The compounds of this invention may contain olefin-like double bonds. When such bonds are present, the compounds of the invention exist as cis and trans configurations and as mixtures thereof.
The present invention also includes isotopically-labeled compounds, which are identical to those recited in general Formulas I or H, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and chlorine, such as 2H, 3H, I3C, 14C, 15N, 18O, I70,31P, 32P, 35S, I8F, and 36C1, respectively. Compounds of the present invention, prodrugs thereof, and pharmaceutically acceptable salts ofsaid compounds or ofsaid prodrugs which contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labeled compounds of the present invention, for example those into which radioactive isotopes such as 3H and 14C are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated, i.e., 3H, and carbon-14, i.e., 14C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium, i.e., 2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labeled compounds of Formula I or II of this invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed herein, e.g., in the Examples, by substituting a readily available isotopically labeled
reagent for a non-isotopically labeled reagent.
A "suitable substituent" is intended to mean a chemically and phannaceutically acceptable funcţional group i.e., a moiety that does not negate the inhibitory activity of the inventive compounds. Such suitable substituents may be routinely selected by those skilled in the art. Illustrative examples of suitable substituents include, but are not limited to halo groups, perfluoroalkyl groups, perfluoroalkoxy groups, alkyl groups, cycloalkyl groups, alkenyl groups, alkynyl groups, hydroxy groups, oxo groups, mercapto groups, alkylthio groups, alkoxy groups, aryl or heteroaryl groups, aryloxy or heteroaryloxy groups, aralkyl or heteroaralkyl groups, aralkoxy or heteroaralkoxy groups, HO—(C ==O)— groups, amino groups, alkyl- and dialkylamino groups, carbamoyl groups, alkylcarbonyl groups, alkoxycarbonyl groups, alkylaminocarbonyl groups dialkylamino carbonyl groups, arylcarbonyl groups, aryloxycarbonyl groups, alkylsulfonyl groups, arylsulfonyl groups and the like.
More specifically, the present invention also relates to a compound having the general Formula I or II
(Figure Removed)
wherein B is either oxygen or sulphur and each "R" is independently defined:
(Figure Removed)
Avitb the requirement that each "R" can never occur only as hydrogen on eifber Formula I or II, and further, that wfthin each "R" independently, any B is either oxygen or sulphur; and "m" is independently either zero or an integer from one to twenty; each X is independently either carbon or nitrogen; and when any X is carbon, then Y is the substituent defined independently for each X as
each Z is independentry either hydrogen, hydroxyl, fhiorine, bromine, iodine, -N,, -OJ, -SR3, -(W.3, -N(R% -*1, or A, and
"n" is independentry either zaro or an integer from one to four, each R1 is independently sefected from hydrogen, (Cj-CjftXcloalkyl, (Cr Ca)alkoxy, (C,-CM)dacyl> phenyl, (C,-C JheteroaryL, (C,-Clo)heterocyclic and (C,-C,»)cycloalkyl; (C,lftJteteroaryM>—, (C,-CJO)heterocyclic-O—, (Cj-Clo)cyclo alkyl-O—, (C.-CJalkyl-S—, (C,-Cj)alkyl-SO2—, (C,-C6)alkyl-NH—SO2—, —NO2, amino, (CgJkyl-amino, [(alkylVamino, (Cj-Calkyl-S02—NH—, (C,-C6)alkyKC=0>—NH—, (C,-C6)alkyKC=O>—[((CrCalkyl)-N]—, phenyl-(0=O)—NI-, phenyHOO)—[((C,-C6)alkyl)-N]— —CN, (Cr C6)aIkyl-(C=O>~-,phenyl-(C=O)—, (Ci-C10)heteroaryl-(C=O)—, (C,-C10)heterocyclic-(OO>—, ((VC10)cycloaIkyl-(OO)—, HO—(0=0)—, (C,— Olkyl-0—(00)—, HjN(00>-(C, -C6)alkyi-NH—(OO)—, [(C,-Cs)alkyl]2 —N—(OO)—, phenyl-NH—(OO>—, phenyl-[((CrC6)alkyl)-N]—(OO)—, (Cj-Cheteroaryl-NH—(OO)—, (C,-C,0)heterocyclic-NH—(OO)—, (C3-

C,0)cycloalkyi-NH—(C=O)—, (C,-Cs)aIkyl-(C=O)—O— and phenyl-(C=O)—O—, wherein each of ti» aforesaid (Cj-Calkyl, phenyl, (Cr Cjheteroaryl, (Cj-CwJbeterocyclic and (Ca-CJcycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C-Ca&yl, (CCJalkenyl, (C2-C6)alkynyl, perhaloCC,-C6)a\kyl, phenyl, (C,-C10)heteroaryl, (C,-C10)heterocyclic, (C3-C10)cycloalkyl, hydroxy, (C-Csilkoxy, pethalcCj-Cjoxy, phenoxy, (C-Ciheteroaryl-O—, (C,-C,0)heterocyclic-O—, (C3-Clft)cycloaikyl-O—, (CpClkyl-S—, (C-Calkyl-SO2~—, (Cj-CUcyl-Ntt—SOj—, —NOj, amino, (C,-C6)alkyl-amino, [(C,-
, (C,-C,)8lkyHC=O)—NH—, (Cr -, phenyl--lH—, phcnyi-, (C,-CJaIkyKC=O)—, phenyKOO)—, (C,-C,»)heteroaryl-(C=O)—, (C,4,)hcterocyclic-{C==O)—, (C3-C)0)cycloalkyl-(G=0>—, HO-OO)—, (C,— Q>dkyl-O—(C=O)—, H(C=O)—(C, -C«)alkyl-NH™(C>*0>—, [(C.-Cttyqj —N—(OO)—, phenyl-NH—.N]—(G=O)—, (C,-Cia)heteroaryl-NH—(C=O>—, (Cr C,0)heterocycUc-NH-—(C=O>—, (—, (C,-C6)alkyl-(C=O)—O— and phenyl-(O=O)—O—; wherein two independently chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which they are attached to form a three to forty membered cyclic, heterocyclic or heteroaryl ring;
each R2 is independently selected from the group consisting of hydrogen, hydroxyl, halo, —N3> —CN, —SH, (R1—N— (R3)—O—, (R3)—S—, (Cr Calkyl, (C2-C4)alkenyl, (Cî-Calkynyl, (C3-C10)cycloalkyl, phenyl, (Cr
C]0)heteroaryl, and (CrC10)hetero-cyclic; wherein each of the aforesaid (Cr C6)alkyl, (C3-C10)cycloalkyl, phenyl, (CrC,0)heteroaryl and (CrC10)heterocyclic substituents may optionally be independently substituted by one to four moieties independently selected from the group consisting of halo, (C,-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(C,-C6)alkyl, phenyl, (C3-C10)cycloalkyl, (Cr C10)heteroaryl, (C,-C10)heterocyclic, formyl, —CN, (C,-C6)alkyl-(C=O)—, phenyl-(C=O>—, HO-(C=O)—, (CrC6)alkyl-O—(C=O>—, (CrC6)alkyl-NH—(C=O)—, [(C,-C6)alkyl]2—-N—(C=O)—, phenyl-NH—(C=O)—, phenyl-[((C ,-C6)alkyl)-N]—(C=O)—, —N02) amino, (C,-C6)alkylamino, [(CrC6)alkyl]2-amino, (C,-C6)alkyl-(C=0)—NH—, (CrC6)alkyl-(C=O)—[((CrCs)alkyl)-N]—, phenyl-(O=O)—NH—, phenyl-(OO)—[((C,-C6)alkyl)-N]—, H2N—(C=O)—NH—, (CrC6)alkyl-HN——NH—, (phenyl-)2N—(C=O)—NH—, phenyl-HN—(OO)—[((CrC6)alkyl)-N]—, (phenyl-N—(C-O)—[((Cr C6)alkyl)-N]—, (CrC6)alkyl-O—(C=O)—NH—, (CrC6)alkyl-O—(C=O>—[((C,-C6)alkyl)-N]—, phenyl-O—(C=O>—NH—, phenyl-O—(OO)—[((C, -C6)alkyl)-N]—, (C,-C6)alkyl-SO2NH—, phenyl-SO2NH—, (C,-C6)alkyl-SO2—, phenyl-SO2—, hydroxy, (CrC6)alkoxy, perhalo(CrC6)alkoxy, phenoxy, (CrC6)alkyl-(C=0)—O—, phenyl-(C=O)—O—, H2N—(C=O)—O—, (CrC6)alkyl-HN—(C=O>—O—, [(C,-C6)alkyl-]2N—(OO)—O—, phenyl-HN—(C=0)—O—, (phenyl-)2 N—(C=0)—O—; wherein when said R2 phenyl contains two adjacent substituents, such substituents may optionally be taken together with the carbon atoms to which they are attached to form a five to six
membered carbocyclic or heterocyclic ring; wherein each ol saia moieuc» containing a phenyl alternative may optionally be substituted by one or two radicals independently selected from the group consisthig of (Cj-C6)alkyl, halo, (C,-C6)alkoxy, perhalo{CrC6)alkyl and perhalo(C,-C6)alkoxy;
each R3 is independently selected from the group consisting of hydrogen, (C3-C20)cycloalkyl, (C,-C20)alkoxy} (CrC20)alkyl, phenyl, (C,-C10)heteroaryl, (Cr Ci0)heterocyclic and (C3-C10)cycloalkyl; wherein each of the aforesaid (C,-Calkyl, phenyl, (C,-C10)heteroaryls (C,-Ci0)heterocyclic and (C3-CM)cycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C1-C6)alkyi, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(C,-C6)alkyl, phenyl, (CrC,0)heteroaryl, (CrC10)heterocyclic, (C3-C]0)cycloalkyl, hydroxy, (CpCalkoxy, perhalo(CrC6)alkoxy, phenoxy, (C,-C10)heteroaryl-O—, (C,-C,0)heterocyclic-O—, (C3-C,„)cycloalkyl-O—, (Cr C6)alkyl-S—, (C,-C6)alkyl-SO2—, (CrC6)alkyl-NH—SO2—, —NO2, amino, (CrC6)alkyl-amhio, [(C,-C6)alkyl32-amino, (Cj-Calkyl-SOj—NH—, (C,-C6)alkyl-(0=0>—NH—, (CrC6)alkyl-(C=O)—[((C,-C6)alkyl>N]—, phenyl-(0-0)—NH—, phenyl-(0=0>—[((CrC6)alkyl)-N]—, —CN, (CrC6)alkyI-(C=O)—, phenyl-(CM))—, (C,-C10)heteroaryl-(C=O)—, (C,-C,0)heterocyclic-(00)—, (C3-CJO)cycloalkyHC=0)—, HO—(OO>—, (C,— C6)alkyl-O—(00)—-, H2N(C=0>—(C, -C6)alkyl-NH—(G=O)—, [(CrCs)alkyl]2 —N~-(C=0)—, phenyl-NH—(C=O>—, phenyl-[((C,-C6)alkyl)-N]—(0=0)—, (CrC,0)heteroaryl-NH—(OO)—, (CrC10)heterocyclic-NH—(C=O)—, (C3-C10)cycloalkyl-NH~(O0>—, (C,-C6)alkyl-(C=O)—O— and phenyl-(00)—O—;
or tbe pfaarmaccutically acceptable satts and prodrugs thereof.
As med heresn, the term "aBcyL," as well as the alkyl moieties of otfaer groups referred to herein (e.g., alkoxy), may be linear or branched (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, secondary-butyl, tertiary-butyl), and they may also be cyclic (e.g., cyclopropyl or cyclobutyl); optionally substituted by l to 3 siritable substituents as defined above such as fluoro, cbioro, trifluoromethyl, (C,-Cg)alkoxy, (Cj-CjftJaryloxy, trifluoromethoxy, difluoromethoxy or (C,-C,5)alkyl. The phrase "each ofsaid alkyT as used herein refers to any of the preceding alkyl moieties within a group such alkoxy, alkenyl or alkylamino. Preferred alkyls închide (C,-C4)e]kyl> most preferably methyl.
As used herein, the ton "cycloalkyF refers to a mono or bicyclic carbocyclic ring (e.g., cyclopropyl, cyclobutyl, cyclopenryl, cydohexyl, cycloheptyl, cyclooctyl, cydononyl, cyckipente&yl, cyciohexeayl, bicyclo[2J2.1Jhcptanyl, bicyclo(3.1)octaoyl «nd bicyclo[5.2.0}noDanyl, etc.); optionally containing 1-2 obubk bonds and optkmally substituted by l to 3 suitable substituents as defined above such as fluoro, chloro, trifluoromethyl, (C,-C6)alkoxy, (C6 -CIO)aryloxy, trifluoromethoxy, difluoromethoxy or (Cj-C6)alkyl. The phrase "each ofsaid alkyl" as used herein refers to any of the preceding alkyl moieties within a group such alkoxy, alkenyl or alkylamino. Preferred cycloalkyls include cyclobutyl, cyclopentyl and cyclohexyL
As used herein, the tem "halogen" or "halo" includes fluoro, chloro, bromo or iodo or fluoride, chloride, bromide or iodide.
As used herein, the term "halo-substituted alkyl" refers to an alkyl radical as descnbed above substituted whh one or more halogens included, but not limited to,
chloromethyl, dichloromethyl, âuoromethyl, difluoromethyl, trifluoromethyl, 2>,2-trichloroethyl, and the like; optionally substituted by l to 3 suitable substituents as defined above such as fluoro, chloco, trifluoromethyl, (C,-C6)alkoxy, (C6-C,0)aryioxy, trifluoromethoxy, difluoromethoxy or (C, -C6)alkyl.
As used herein, the tenn "alkenyl" means straight or branched chain unsaturated radicals of 2 to 6 carbon atoms, including, but not limited to ethenyl, l -propenyl, 2-propenyl (allyl), iso-propenyl, 2-methyl-1 -propenyl, 1-butenyl, 2-butenyl, and the like; optionally substituted by l to 3 suitable substituents as defined above such as fluoro, chloco, trifluoromemyl, (CfCalkoxy, (C6-C10)aryloxy, trifluoromethoxy, difluoromethoxy or (C,-Cj)alkyl.
As used herein, the tenn "(CfCăkyayV îs used herein to mean straight or branched hydrocarbon chain radicals having one tripk bond mchiding, but not limited to, ethynyL, propynyl, bntynyl, and ihe Kke; optionally substituted by l to 3 suitable substituents as defined above such as fluoro, chloro, trifluoromemyl, (C,-Cg)alkoxy, (C6-C10)aryloxy, trifluoromethoxy, difluoromemoxy or (CVCalkyl.
As used herein, the tenn "carbonyl" or "(C=O)" (as used in phrases such as alkylcarbonyl, alkyl-(OO)— or alkoxycarbonyl) refers to the joinder of the >C=O moiety to a second moiety such as an alkyl or amino group (i.e., an amido group). Alkoxycarbonylamino (i.e., alkoxy(OO)—NH —) refers to an alkyl carbamate group. The carbonyl group is also equivalently defined herein as (C=O). Alkylcarbonylamino refers to groups such as acetamide.
As used herein, the term "phenyl-JXCj-CgJalkyiyN]—(C=O)—," refers to a disubstituted amide group of the formula:

(Figure Removed)
As used herein, the term "aryl" means aromatic radicals such as phenyl, naphthyl, tetrahydronaphthyl, indanyl and the like; optionally substituted by l to 3 suitable substituents as defined above such as fluoro, chloro, trifluoromethyl, (C,-C6)alkoxy, (C6-CIO)aryloxy, trifluoromethoxy, difluoromethoxy or (C,-C6)alkyl.
As used herein, the term "heteroaryl" refers to an aromatic heterocyclic group with at least one heteroatom selected from O, S and N in the ring. hi addition to said heteroatom, the aromatic group may optionally have up to four N atoms in the ring. For example, heteroaryl group includes pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl, furyl, imidazolyl, pyrrolyl, oxazolyl (e.g., 1,3-oxazolyl, 1,2-oxazolyl), thiazolyl (e.g., 1,2-thiazolyl, 1,3-thiazolyl), pyrazolyl, tetrazolyl, triazolyl (e.g., 1,2,3-triazolyl, 1,2,4-triazolyl), oxadiazolyl (e.g., 1,2,3-oxadiazolyl), thiadiazolyl (e.g., 1,3,4-thiadiazolyl), quinolyl, isoquinolyl, benzothienyl, benzofuryl, indolyl, and the like; optionally substituted by l to 3 suitable substituents as defined above such as fluoro, chloro, trifluoromethyl, (Cj-C6)alkoxy, (C6-C,0)aryloxy, trifluoromethoxy, difluoromethoxy or (C,-C6)alkyl.
The term "heterocyclic" as used herein refers to a cyclic group containing l-9 carbon atoms and 1-4 hetero atoms selected from N, O, S or NR1. Examples of such rings include azetidinyl, tetrahydrofuranyl, imidazolidinyl, pyrrolidinyl,
piperidinyl, piperazinyl, oxazolidinyl, thiazolidinyl, pyrazolidinyl, thiomorpholinyl, tetrahydrothiazinyl, teţrahydrothiadiazinyl, morpholinyl, oxetanyl, tetrahydrodiazinyl, oxazinyl, oxathiazinyl, indolinyl, isoindolinyl, quinuclidhiyl, chromanyl, isochromanyl, benzoxazinyl and the like. Examples of such monocyclic saturated or partially saturated ring systems are tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, imidazolidin-1-yl, imidazolidin-2-yl, imidazolidin-4-yl, pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl, piperazin-1-yl, piperazin-2-yl, piperazin-3-yl, l,3-oxazolidin-3-yl, isothiazolidine, l,3-thiazolidin-3-yls l,2-pyrazolidhi-2-yl) 1,3-pyrazolidin-l-yl, thiomorpholinyl, l,2-tetrahydrothiazin-2-yl, l,3-tetrahydrothiazin-3-yl, teţrahydrothiadiazinyl, morpholinyl, l,2-tetrahydrodiazin-2-yl, 1,3-tetrahydrodiazin-1-yl, l,4-oxazin-2-yl, l ,2,5-oxathiazin-4-yl and the like; optionally substituted by l to 3 suitable substituents as defined above such as fluoro, chloro, tnfluoromethyl, (Cj-C6)alkoxy, (C6-C10)aryloxy, trifluoromethoxy, difluoromethoxy or (C,-C6)alkyl.
Another embodiment of the present invention includes those compounds having a chemical structure within one of the following two formulas:
(Figure Removed)
wherein R and B are defined as in general Formula I and n above with the
exception that at least one R in each above chemical structure formula contains one of the two following chemical sub-structures
(Figure Removed)
and Ar is either one of tibe foQowing eight chemical sub-structures
(Figure Removed)
or Ar is defîned K one of the fbOowing three chemical sub-structures(Figure Removed)
wherein each X is independently either carbon or nitrogen; and when any X is carbon, then Y is the substituent defined independently for each X as
(Figure Removed)
each Z is independentry either hydrogen, hydroxyl, fluorine, bromine, iodine, -N3, -CN, -SR3, -OR3, -NCR1, "n" is independently either zero or an
r integer trom one to four, and R' and R3 are defined as in general Formula I or n. A preferred embodiment bere is wherein B is oxygen and/or from the R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-C)cycloalkyl, (C,-C2B)alkoxy, (C,-CM)alkyl, phenyl, (C,-C10)heteroaryl, (Cr Cta)heteroeyclic and (C3-Cto)cycloalkyl; wherein each of the aforesaid (C,-Calkyl, phenyl, (C,-Cia)heteroaryl, (C,-C,0)heterocyclic and (Cj-Ccycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (Cl-C6)alkyl, (C2-Cs)alkenyl, (2-C6)alkynyl, ţtateWC}-Cd*&yl* phenyl» (C,-Cla)heteroaryl, (C,-Cjn)heterocyclic, (C,-C10)cycloalkyl, hydroxy, (Cj-CJaDcoxy, perhalo(C1-C -, (Cj-CJcycloalkyl-O—, (C,-
CJalkyl-S—; wherein two indepeadentry chosen R1 allcyl-containmg groups may be taken together witti any nitrogen atom to whkhtheyareatrachedtoform athree tofortynMrabered,cychheerocychorheteToaryl Still more preferred are when R and R1 are defined as independently selected from the group consisting of hydrogen, (-Cjloaikyl, (C,-C20)alkoxy, (C-Calkyl, phenyl, (Cr C10)heteroaryl, (C,-Clo)heterocyclic and (C3-C,0)cycloalkyl; wherein each of the aforesaid (Ci-C2o)alkyl, phenyl, (C,-C10)heteroaryl, (C,-Ci„)heterocyclic and (C3-C20)cycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C,-C6)alkyl, (C2-C6)alkenyl, (Cj-CJalkynyl, perhalo(C,-C6)alkyl, phenyl, (CrCto)heteroaryl, (Cr C10)heterocyclic, (C3-C10)cycloalkyl, hydroxy, and (CrC(s)alkoxy. Even still more preferred is when R and R1 are defined as independently selected from the group consisting of hydrogen, (Q-£w)cycloalkyl, (Cj-Cj0)alkoxy, (Ci-Clo)alkyl, phenyl,
(CrC,0)heteroaryl, (C,-C10)heterocyclic and (C3-C10)cycloalkyl. Most preferred is when R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-C6)cycloalkyl, (C,-C6)alkoxy, and (C,-C6)alkyl.
Another embodiment of the present invention includes those compounds having a chemical structure within one of the following two formulas:

(Figure Removed)


wherein Ar, R, B and R1 are as defined in general Formula I and II above. A preferred embodiment here is wherein B is oxygen and/or R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-C20)cycloalkyl, (C1-C20)alkoxy, (C,-CM)alkyl, phenyl, (C,-C10)heteroaryl, (C,-C,0)heterocyclic and (C3-C10)cycloalkyl; wherein each of the aforesaid (C,-C20)alkyl, phenyl, (C,-C10)heteroaryl, (C,-C,0)heterocyclic and (C3-C2o)cycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C,-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(C,-C6)alkyl, phenyl, (C,-C10)heteroaryl, (C,-C,0)heterocyclic, (C3-C,0)cycloalkyl, hydroxy, (C,-C6)alkoxy, perhalo(C,-C6)alkoxy, phenoxy, (C,-Ci0)heteroaryl-O — , (C,-C,0)heterocyclic-0 — , (C3-C10)cycloalkyl-O — , (C,-C6)alkyl-S — ; wherein two independently chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which they are attached to form a three to forty membered, cyclic, heterocyclic or heteroaryi ring. Still more preferred are when R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-
C20)cycloalkyl, (CrC20)alkoxy, (CrC20)alkyl, phenyl, (CrC10)heteroaryl, (Cr C]0)heterocyclic and (C3-C10)cycloalkyl; wherein each of the aforesaid (C,-C20)alkyl, phenyl, (CrC10)heteroaryl, (C,-C10)heterocyclic and (C3-C20)cycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C,-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(C,-C6)alkyl, phenyl, (C,-Ci0)heteroaryl, (CrC10)heterocyclic, (C3-C10)cycloalkyl, hydroxy, and (C,-C6)alkoxy. Even still more preferred is when R and R1 are defîned as independently selected from the group consisting of hydrogen, (C3-C10)cycloalkyl, (CrC10)alkoxy, (C,-C10)alkyl, phenyl, (Cr C,0)heteroaryl, (C,-Ci0)heterocyclic and (C3-C10)cycloalkyl. Most preferred is when R and R1 are defîned as independently selected from the group consisting of hydrogen, (C3-C6)cycloalkyl, (C,-C6)alkoxy, and (C,-C6)alkyl.
Another embodiment of the present invention includes those compounds having a chemical structure within one of the following two formulas:

(Figure Removed)
wherein R and B are as defined in general Formula I or II above, and Ar is either one of the following eight chemical sub-structures
(Figure Removed)


or Ar is defined as one of tbe following three chemical sub-structures

(Figure Removed)
wherin each X is independentty either carbon or nitrogen; and wfaen any X is carbon, then Y is the subttitoent defined independently for each X as


(Figure Removed)
each Z is independently either hydrogen, hydroxyl, fluorine, bromine, iodine, -N3, -CN, -SR3, -OR3, -NCR1, "n" is independently either zero or an integer from one to four; and R1 and R3 are defined as in general Formula I or n. A preferred embodiment bere is wherein B is oxygen and/or R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-Ccycloalkyl, (Ct-C2o)alkoxy, (C.-Calkyl, phenyl, (C,-C10)heteroaryl, (Cr C10)heterocyclic and (C3-C)8)cycloalkyl; erein each of the aforesaid (Cr
l, (C,-C10)heteroaryl, (C,-Cw)heterocycIic and substituents may optionalry be substituted by one to four moieties independently selected firom the group consisting of halo, (Cj-CdJalkyl, ((-(alkenyl, (C2-Clkynyl, peAalo-CJalkyl, phenyl, (C,-C10)heteroaryl, (C,-C,0)heterocyclic, (C.-CMOcycloalkyl, hydroxy, (C,-Cs)alkoxy, pernaloO-Csoxy, phenoxy, (Cr C,o)heteroaryl-O—, (C, Jheterocyclic-O—, (C3-Cw)cycloalkyl-O—, (Cr Ca)alkyl-S—; wherein two independently chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which they are attached to form a three to forty membered, cyclic, heterocyclic or heteroaryl ring. Still more preferred is when R and R1 are defîned as tndependentry selected firom mc group consisting of hydrogen, (CfCcyM&y (C,-C,Jdkoaiy, (C1-CM)rfkyi, phenyl (Cr CJheterogul, (C,-€,Jheterocyclic and ((-CJcycloalkyi; wherem each of the aforesaid (C,-Cn)aIkyi, phenyi (C,-C Jheteroaryl, (C,-CJheterocyclic and (C,-Ca)cycloalkyl substituents may oţtkmalrybesutenHatedby one to four moieties independently selected firom the group consisting of halo, (C,-C Another embodunent of the present invention includes those compounds having a chemical structure within one of the following two fonnulas:

(Figure Removed)
wherein Ar, R, B and R1 are as defined in general Formula I and II above. A
preferred embodiment here is wherein B is oxygen and/or R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-C2())cycloalkyl, (C1-C20)alkoxy, (C,-C20)alkyl, phenyl, (C,-C10)heteroaryl, (CrC10)heterocyclic and (C3-C10)cycloalkyl; wherem each of the aforesaid (C,-C20)alkyl, phenyl, (C,-C10)heteroaryl, (CrC10)heterocychc and (C3-C20)cycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C,-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(C,-C6)alkyl, phenyl, (CrC10)heteroaryl, (C,-C10)heterocyclic, (C3-C10)cycloalkyl, hydroxy, (C,-C6)alkoxy, perhalo(C,-C6)alkoxy, phenoxy, (CrC,0)heteroaryl-O—, (CrC10)heterocyclic-O—, (C3-C,0)cycloalkyl-O—, (C,-C6)alkyl-S—; wherein two independently chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which they are attached to form a three to forty membered, cyclic, heterocyclic or heteroaryl ring. Still more preferred are when R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-C20)cycloalkyl, (C,-C20)alkoxy, (CrC20)alkyl, phenyl, (CrC10)heteroaryl, (Cr C10)heterocyclic and (C3-C10)cycloalkyl; wherein each of the aforesaid (Cr C20)alkyl, phenyl, (CrC10)heteroaryl, (C,-C!0)heterocyclic and (C3-C20)cycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (Cj-Calkyl, (C2-C6)alkenyl, (C2-
C6)alkynyl, perhalo(CrC6)aikyl, phenyl, (CrC,0)heteroaryl, (C,-C,0)heterocyclic, , (C3-C10)cycloalkyl, hydroxy, and (CrC6)alkoxy. Even still more preferred is when R and R! are defined as independently selected from the group consisting of hydrogen, (C3-C10)cycloalkyl, (C,-C10)alkoxy, (C,-C10)aIkyI, phenyl, (Cr C10)heteroaryl, (C,-Clo)heterocyclic and (C3-C,0)cycloalkyl. Most preferred is when R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-C6)cycloalkyl, (C,-C6)alkoxy, and (CrC6)alkyl.
Another embodiment of the present invention includes those compounds having a chemîcal structure within one of the following two formulas
(Figure Removed)

wherein R is defined as in general Formula I and n above and Ar is either one of the following eight chemical sub-structures
(Figure Removed)



or Ar is defined as ane of the foilowing three cbemical sub-structures

(Figure Removed)


wherein each X is independently either carbon or nitrogen; and when any X is carbon, then Y is the substituent defined independently for each X as

(Figure Removed)
each Z is independently either hydrogen, hydroxyl, ftuorine, bromine, k»dine,-ls,"CN,-SR3>-OR3,l(Rl)2> "n" is independently either zero or an integer firom one to four; and R1 and R' are defined as in general Formula I or n. A preferred embodiment hore is wherein B is oxygen and/or R and R1 are defined as independently setected from the group consisting of hydrogen, (C,-CaOcycloalkyl, (Ci-CM)attoxy, (Cj-Calkyl, phenyl, (C,-C,heteroaryl, (Cr C10)heterocyclic and (C3-Clft)cycloalkyl; wherein each of the aforesaid (C,-Cao)alkyl, phenyl, (C,-Cloieteroaryl, (C,-C10)heterocycUc and (Cj-Ccycloalkyl substituents may optionally be substituted by one to four moieties independently selected fiom the group consisting of halo, (C,-C6)alkyl, (C2-C6)alkenyl, (C-Cs)alkynyl, perhalo(C|-Cs)alkyl, phenyl, (Ci-C10)heteroaryl, (Ci-C10)heterocyclic, (C3-C,0)cycloalkyl, hydroxy, (Calkoxy, perhaloCCj-Cjoxy, phenoxy, (C,-Ci0)heteroaryl-O—, (C,-C10)heterocyclic-O—, (C3-C10)cycloalkyl-0—, (Cr C6)alkyl-S—; viierein two independently chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which they are attached to form a three
to forty membered, cyclic, beterocyclic or heteroaryl ring. Still more preferred are when R and R1 are defined as independently selected firom tbe group consisting of hydrogen, -Ccycloeflcyl, (C.-Calkoxy, (C.-Calkyl, phenyl, (Cr CIO)heteroaryl, (C,w)heterocyclic and (C3-C10)cycloalkyl; wherein each of the aforesaid (Cj-Cjo)6!. phenyl, (C,-C|0)heteroaryl, (Ci-ClftJheterocyclic and (C,-C2o)cycloalkyi substituents may optionally be substituted by one to four moieties independently selected firom the group consisting of halo, (C,-C6)alkyl, (Cj-C>lkenyl, (CfCJOkyayl, perhaloCCi-CiJalkyl, phenyl, (C,-Cie)heteroaryl, (Cr C,0)heterocyclic, (C3-C10)cycloalkyl, hydroxy, and (C,-CJalkoxy. Even still more preferred is when R and R1 are defined as independently selected firom the group consisting of hydrogen, (C3-CM)cyck>«lkyl, (C.-CJalkoxy, (C,-C„)alkyl, phenyl, (C,)0)hcteroaryl,(C,Jbctm>cyclkand(Cyck)«akyL Mostprefenedis when R and R1 are defined as independently setected firom the group consisting of hydrogen, (Cj-CJcydoalkyl, (C,-CJalkoxy, and (C,-CJalkyL
Another embodhnent of the present invention inchides those compounds having a chemical structure within one of the following two formulas:

(Figure Removed)

wherein Ar, R, B and R1 are as defined in general Formula I and n above. A preferred embodhnent bere is wherein B is oxygen and/or R and R1 are defined as independently selected firom the group consisting of hydrogen, (Cs-Cycloalkyl, (C,-C20)alkoxy) (-Cjodkyl, phenyl, (C,-C,0)heteroaryl, (CrC,0)heterocyclic and
(C,-Cw)cycloalkyl; wherein each of me aforesaid (Cj-Calkyl, phenyl, (C,-C,e)heteroaryl, (C,-Cie)hetorocycIic ana (Cy-Ccycloalkyl substituents may optionally be substituted by oue to four moieties independently selected from the group consisting of halo, (C.-CjJalkyl, (CVCeny!, (C2-C6)alkynyl> perhalo(Cr CIkyl, phenyl, (C.Kjbeteroaryl, (C1-C10)heterocycUc> (C, Jcycloalkyl, hydroxy, (Cj-Cjlkoxy, pcrhalo(C,-C6)alkoxy, phenoxy, (C,-C,0ieteroaryI-O—, (C,-Cie)hetero Another embodiment of the present invention includes those compounds
baving a chemical structure within one of the following two formulas:

(Figure Removed)
wherein R and B are as defined in general Formula I or n above, Ar is either one of the following eight chemical sub-structures
(Figure Removed)
or Ar is defined as one of the following three chemical sub-structures
(Figure Removed)
\wherein each X is independently either carbon or nitrogen; and when any X is carbon, then Y is the substituent defined independently for each X as
(Figure Removed)
each Z is independently either hydrogen, hydroxyl, fluorine, bromine, lodine, ~N3, -CN, -SR3, ~OR3, -NCR1) "n" is independently either zero or an integer from one to four; and R1 and R9 are defmed as in general Formula I or n. A preferred embodiment here is wherein B is oxygen and/or R and R* are defmed as independently selected from the group consisting of hydrogen, (Cj-QaOcycloalkyl, (C.-Calkoxy, (C.-Calkyl, phenyl, (C,-C10)heteroaryl, (Cr C10)heterocyclic and (C3-C,0)cycloallcyl; wherein each of the aforesaid (C,-CaOalkyl, phenyl, (Cj-CjaJheteroaryl, (C,-C10)heterocyclic and (Ca-CfflJcycloalkyl substituents may optionaUy be substituted by one to four moieties independently selected from the group consisting of halo, (Ci-Cf)alkyl, (C2-C&)alkenyl> (Q" CJalkynyl, perhalo(CI-C6)alkyU phenyl, (C.-CJheteroaryl, (CJheterocyclic,
, (C.-CJrikoxy, perhaloCC.-Clkoxy, phenoxy, (Cr , (C,-C Jhcterocyclic-O— , (Ca-CJcycloâlkyl-O— , (C,-
Cj)alkyl-S — ; wherein rwo independeitfry chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which Aey are attached to form a three to forty membered, cyclic, heterocycUc or heteroaryl ring. Still more preferred is when R and R1 are defmed as independently selected from the group consisting of hydrogen, (Cs-Ccycloalkyl, (C,-C20)alkoxy, (C-Calkyl, phenyl, (Cr C10)heteroaryl, (C,-C)0)heterocyclic and (C3-C10)cycloalkyl; iierein each of the aforesaid (C,-C2o)alkyl, phenyl, (C,-Clo)heteroaryl, (Cj-CiaJheterocyclic and (C3-C2o)cycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C,-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(C1-C6)alkyl, phenyl, (CrC,0)heteroaryl, (Cr C10)heterocyclic, (C3-C10)cycloalkyl, hydroxy, and (C,-C6)alkoxy. Even still more
preferred is when R and R1 are defined as independently selected from the group consisting of hydrogen, (C<:>alkyl, (C,-C,0)alkoxy> (Ci-C,„)alkyl, phenyl, (Ci-Cheteroaryl, (C,-C,0)beteiocyclic and (C3-C,0)cycloalkyl. Most preferred is when R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-C6)cycloalkyl, (C.-Clkoxy, and (C,-C6)alkyl.
Another embodiment of the present invention includes those compounds having a chemical structure wfthin one of the following two formulas:

(Figure Removed)
wherein Ar» R, B and R1 axe as defined in general Formula I and U above. A preferred embodiment bere is wherein B is oxygen and/or R and R1 are defined as independently selected from the group consisting of hydrogen, (Cj-Ccycloalkyl, (CVCalkoxy, (C-Calkyl, phenyl, (C,-C10)heteroaryl, (C,-C,0)heterocyclic and
i
(C3-C10)cycloalkyl; wherein each of the aforesaid (Cj-Calkyi, phenyl, (Cj-C10)heteroaryl, (C|-C10)heterocyclic and (Cj-Cycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C-Calkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(Cr C6)alkyl, phenyl, (C,-C10)heteroaryl, (C,-C10)heterocycUc, (C3-C,0)cycloalkyl, hydroxy, (C,-C6)alkoxy, perhalo(C,-C6)alkoxy, phenoxy, (CrC10)heteroaryl-0—, (C,-C10)heterocyclic-0—, (C3-C10)cycloalkyl-0—, (Cj-Calkyl-S—; wherein two
independently chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which they are attached to form a three to forty membered cyclic, heterocyclic or heteroaryl ring. Still more preferred is when R and R1 are defined as independently selected from Ine group consisting of hydrogen, (C3-CMJcycloalkyi, (C.-Calkoxy, (Cj-CjoJalkyl, phenyl, (C,-C,0)heteroaryl, (Cr C)0)heterocyclic and (C,-C,0)cycloalkyl; wherein each of the aforesaid (C,-Caalkyl, phenyU (C,-C,,ieteroaryl, (C,-C,0)heterocyclic and (C-Ccycloalkyl substituents may optionalry be substituted by one to four moieties independently selected from the group consisting of halo, (C1-C6)alkyl> (Cj-Calkenyl, (C2-CJalkynyl, p«halo(C1-CJalkyl, phenyl, (C,-CM)heteroaryl, (C,-C,o)heterocyclic, ((„toalkyl, hydro, and (C,->lkoxy. Evtn stiO more preferred is R and R1 aţe defined as mdependentry selected from Ae group consisting
Most preferred is when R and R1 are defined as mdependenây selected from tiie group consisting of hydrogen, (Cj-Ccycloalkyl, (CJalkoxy, and (Cj-Clkyl.
Another embodiment of the present invention includes those compounds having a chemical structure witnin one of the following two formulas:

(Figure Removed)
wherein R is defined as in general Formula l and n above and Ar is either one of the following eight chemical sub-structures

(Figure Removed)
wherein each X is independendy eitfier carbon or nitrogen; and wfaea any X is carbon, Aen Y is Ihe substituent defined independentry fer each X as
or Ar is defined as one of the following three chemical sub-structures
T
each Z is independently either hydrogen, hydroxyl, fluorine, bromine, iodine, -N3, -CN, -SR3, -OR3, -NR1, "n" is independently either zero or an integer from one to four; and R1 and R3 are defined as in general Formula I or H. A preferred embodiment bere is wherein B is oxygen and/or R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-Csoycloalkyl, (C,-C20)alkoxy, (Q-Calkyl, phenyl, (Cj-C,0)heteroaryl, (Cr C10)heterocyclic and (C3-C,0)cycloalkyl; wherein each of the aforesaid (Cr

CgOalkyl, phenyl, (C,-CIO)heteroaryl, (C,-C,0)hete3-ocyclic and (Cj-Ccycloalkyl substituents may optionally be substituted by oue to four moieties independently selected from the group consisting of halo, (C,-C6)alkyl, (C2-C6)alkaiyl, (Cj-C6)alkynyl, perijaloCC-Cj)!, phenyl, (C,-Cia)heteroaryl, (CrC,0)heterocyclic, (C3-Clb)cycloalkyl, hydroxy, (C-CJalkoxy, perhakKQ-CsJalkoxy» phenoxy, (C,-C,0)heteroaryl-O — , (C,-Clo)heterocycHc-O — , (C3-C10)cycloalkyl-O — , (Cr C6)alkyl-S — ; \vherein two mdependently chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which tfaey are attached to form a three to forty membered cyclic heterocyclic or heteroaryl ring. Still more preferred is when R and R1 are defined as independently selected from Ine group consisting of hydrogen, (C3-CM)cyeloaIkyl> (C,-CM)rikoxy, (CfCy&kyl, phenyi, (Cr CMteroaryU (CJheterocyclic aad (C3-C10)cycloelkyl; whercin each of the aforesaid (C.-CalkyU phenyU (C.-CJbeteroaryl, (C,-C)t)hetcrocyclic and (C,-
mH»y «fHMialiy ttt>. MihatHiiteH hy ong tn fam- mniptire
independently selected from the group consisting of halo, (C,-Cc)alkyl, C6)alkenyl> (Cj-Clkynyl, perhalo(C,-C6)alkyl, phenyi, (C,-C10)heteroaryl, (Cr C10)heterocyclic, (C3-C10)cycloalkyl, hydroxy, and (C,-C6)alkoxy. Even still more preferred is when R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-C10)cycloalkyl, (C,-C,0)alkoxy, (C,-C,0)alkyl, phenyi, (CrC,0)heteroaryl, (C,-C,0)heterocyclic and (C3-C10)cycloalkyL Most preferred is when R and R1 are defined as independently selected from the group consisting of hydrogen, (C,-C6)cycloalkyl, (Calkoxy, and (C1-C8)alkyL
Other embodiments of the present invention reiate to those compounds described above or listed in TABLE I attached below, either as to the individual

compound itsell or in a composition, or the process of making or the use thereof in methods according to the invention. In each of the compovmds Usted in TABLE I below, any hydrogen may be replaced by the substituent Rx which is a (Q-Calkyl, (Cî-CJalkenyl, (Cj-C>IkynyI, phenyl, (C,-C,0)heteroaryl, (C,-C10)heterocycUc or (C3-C10)cycloalkyl substituent. Otber embodiments of the invention are related to the specific subgenuses Usted in TABLE L hi these subgenuses, any hydrogen can also be replaced by an R substituent.
TABLE 1
(Table Removed)
The compounds of tbe present invention have utility in pharmacological compositions for the treatment and prevention of many diseases and disordcrs characterized by a MTF response, wbereby MIF is released from cellular sources and MIF production is enhanced. A rompound of the invention can be administered to a hunaan pstient by itself or in phannaceutical compositions where it is mixed with suitable carriers or excipients at doses to treat or ameliorate various conditions characterized by MIF release. A therapeutically effective dose may refer to that amount of the compound sufficient to inhibit MIF tautomerase activity and MIF bioactivity, it being understood tfaat such inhibition may occur at different concentrations such that a person skilled in the art could determine the required dosage of coinpoundtombibft the target MIF activity. Therapeuticalty effective doses may be administered alone or as «djunctive thenqy in combination wiui oAer treatments, such as steroidal or non-steroidal anti-infiammatoTy agents, or ano-tumor agents. T
mrtant applirarfwn tn«v be fbnnd in ţ>pjjr|pton's PhermaCCUtical
Sciences. Mack Publishing Co., Baston, PA, latest addition.
Suitable routes of administration may, for example, include oral, rectal, transmucosal, buccal, intravaginal, or intestinal administration; parenteral delivery, including intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, or intraocular injections, and optionally in a depot or sustained release formulation. Furthermore, one may administer a compound of the present invention in a targeted drug delivery system, for example in a liposome.
The pharmaceutkal compositions and compounds of the present invention
may be manufactured in a marmer tbat is itself known, e.g,, by means of convenţional mixing, dissorving, dragee-making, levitating, emulsifying, encapsulating, entrapping, or ryophilizing processes. Pharmaceutical compositions for use in accordance with the present invention thus may be formulated in convenţional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries that facilitate processing of the active compounds into preparations, which can be used phannaceutically. Proper formulation is dependent upon the route of administration chosen.
Any combination of one or more compounds of Fonnulas I, n, salts, prodrugs, metabolites, isotopicalry-labeled compounds, tautomers, isomers, and/or atropisomers is possible in tfae composition.
For injection, me compounds of mc invention may be formulated in aqueous sorutions, preferabty m physioiogicalry compatible buffers, such as Hank's soliition,Ringer'ssohitkm,OTphysk4ogical saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are knovm in the art
For oral administration, the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers well known to those in the art Such carriers enable the compounds of the invention to be formulated as tablete, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the Uke, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained by combining the compound with a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to
obtain tablete or dragec cores. Suitable excipients are, in particular, filiere such as sugars, racluding lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyi cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or poryvmylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a şah thereof such as sodium alginate.
Dragee cores are provided whh suitable coatings. For this purpose, concentrated sugar sohitions may be used, wbich may optionally contain gum arabic, talc, poryvinyl pyrrolidone, carbopol gel, poryethylene glycol, and/or titanium dioxide, lacquer sohrtions, and suitaMe organic solvente or solvent mixtures. Dyestu£& or rÂginents may be added to the tablete or drageecx) idenrificatknortodiaracterizedifiereitfcombm
capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as gh/cerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. AII formulations for oral administration should be in dosages suitable for such administration.
For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.
For administration by inhalation, the compounds for use according to the present invention are conveniently delivered in the fbrm of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propeilant, e.g., dichlorodifluoromemane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount Capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder basc such as lactose or starch.
The compounds may be formulated for parenteral administration by
mjectkm,e.g.>bybohBffijertk»orcoiaini Formulations for
injection may be ptesented in unit dosage farm, e.g., in ampoules or m muM-dose containers, witn an added preservative. The compositions may take soch forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabiliging and/or dispersing agents.
Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oii, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as polyionic block (co)polymer, sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the
fe compounds to aliow for the prcparation of Mghly concentrated solutions, e.g., polyiooic block (co)potymeTS.
Ahemativery, the active ingredient may be in powder fonn for constitution with a suitable vehicle, e.g., sterile pyrogen-iree water, before use.
The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing convenţional suppository bases such as cocoa butter or other glycerides.
In addition to fhe formulations described previousty, the compounds may also be fonnulated as a depot preparation. Such long acting formulations may be adnuiiisteredbyimplanta!ioa(fo
by intramuscular injectim. ThtB,ibrc3umrucompo«mdsinaybefoqrmulated wiatabiepoiymericorhydropfaobicmrtcnils (JbrexampteasmemubkMinan aciccytabkiCcykMCTrhimgr.iesiporasspMringly sohib example, as a spuiny soluble şah.
Liposomes and emulsions are well known examples of delivery vebicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethylsulfoxide also may be employed, although usually at me cost of greater toxicity. Additionally, the compounds may be delivered using a sustained-release system,
i
such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent Various forms of sustained-release materials have been established and are well known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biologica! stability of the merapeutic reagent, additional strategies for protein stabilization
may be empioyed,
The phannaceutical compositions also may comprise suitable solid- or gel-phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatra, and polymers such as polyethylene glycols.
Many of the compounds of the invention identified as inhibitors of MD? activity may be provided as salts with pharmaceutically compatible counterions. Pharmaceutically compatible salts may be fonned with many acids, including but not Umited to hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc.; or bases. Salts tend to be morc sohible in aqneous or other protonic solvente thanare the correspondmg fiee basc forme. ExMmpks of pharmaceuricalry acceptaMe sahs, carners or exciptents are weU known to those skillcd in the art and can be found, fer eyampif- în pfgptpytnm'* PhfflmPTţutical Sciences. IMi EditioiL AJL GennaroT Ed.,M»ckPuWishmgCoE«ston>PA(1990). Such salts include, but are not limited to, sodium, potassium, fimium, calcium, magnesium, iron, zinc, hydrochloridc, hydrobromide, hydroiodide, acetate, citrate, tartrate and maleate salts, and the like.
Phannaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained hi an effective amount to achieve their Lotended purpose. More specifically, a therapeutically effective amount means an amount effective to prevent or inhibit development or progression of a disease characterized by MU7 release and production in the subject being treated. Determination of the effective amounts is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided
herein.
For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from tautomerase inhibition assays and cell culture assays. Such hrformation can be used to more accurately determine useful doses in humans Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical, pharmacological, and toxicologica! procedures in cell cuftures or experimental animals, e,g., for determining the LD,,, (the dose lethal to 50% of the population) and the ED» (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is ine therapeutic index and it can be expressed as the ratiobetweenLDg, andED». Compcwdsthatcndubithightherapeurki
EDjLDg) are preferred The data obtamed from cell culture assays or animal
studiescanbettsedmformi&tângarnrfdosagefCT The
dosage of soch compounds Hes prefcrabry wnirin a range of circulating coricentioris that închide u EDM wrth Uttle or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patienf s condition. (See
Chapter. l page 1).
Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety which are sufficient to maintain the desired modulating effects, or minimal effective concentration (MEC). The MEC will vary for each compound but can be estimated from in vitro data; e.g., the concentration necessary

to acbieve a 50-90% inhibition of MIF activity. Dosages necessary to acbieve the MEC will depend on individual characteristics and route of administration. However, HPLC assays, bioassays or immunoassays can be used to determine plasma concentrations.
Dosage intervals can also be determined using the MEC value. Compounds should be administercd using a regimen that maintains plasma levels above the MEC for 1-90% of ti» time, preferably between 30-90% and most preferably between 50-90%. These ranges include l, 2,3,4,5,6,7,8,9,10,20,30,40, 50, 60,70,80,90,99, and any combinaticm thereof.
The active ingredient may be present in a phannaceutical composition in an amount ranging trona 0.1 to 995% by wcight These ranges include 0.1,0 .5, 0.9, l, 2,3,4,5,6,7,8,9,10,20,30,40,50,60,70,80,90,99,99.5,99.9% by wcight
»
and any combinstion thereof.
fflcvses of tocai admimstntion for
organ or tissue, or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.
The amount of composition administered will, of course, be dependent on the subject being treated, on the subjecf s weight, on the subject's age, on the severity of the affliction, on the manner of administration, and on the judgment of the prescribing physician.
The compositions may, if desired, be presented in a pack or dispenser device that may contain one or more unit dosage forms containing the active ingredient The pack may for example comprise metal or plastic foii, such as a blister pack. The pack or dispenser device may be accompanied by instructions for
administration. Compositions comprising a compound of the invention fonnulated in a compatible pbarmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.
The compounds of Formulas I or n, or a pharmaceutically acceptable salt thereof can be used in the mamrfacture of a medicament for the prophylactic or therapeutic treatment of any disease state in a human, or omer mammal, wbich is exacerbated or caused by excessive or unregulated cytokine production by such mammal's cells, such as but not limhed to monocytes and/or macrophages.
The enzyme activity (tautomerase) of MD7 and the substrates it accepte provideanenzyinarkactivityassayfbr
bindtoMIF and disrupt hs biological activity. The present invention provides methods of usc for the compounds in a gem» of such compounds having isoxazohne structures.
The present inventkmfurther provides a phannaceutical composition comprising the isoxazoline compound, or a phannaceuticaUy acceptable salt thereof, and a pharmaceutically acceptable carrier or diluant, wherein the composition comprises an effectrve amount of the compound of the above formula.
The present invention also provides a pharmaceutical composition comprising a compound having an isoxazoline or isoxazoline-related moiety, and a pharmaceutically acceptable carrier, wherein the compound forms a stable interaction with at least one amino acid residue of a MTF protein.
The present invention provides a method for treating inflammatory disorders (including, but not limited to, arthritis, proliferative vascular disease, ARDS (acute respiratory distress syndrome), cytokine-mediated toxicity, sepsis,
septic shock, psoriasis, interleukin-2 toxicity, asthma, MIF-mediated conditions, autoimmune disorders (inchiding but not limited to rheumatoid arthritis, insulin-dependent diabetes, multiple sclerosis, graft versus nost disease, lupus syndromes), tumor growth or angiogenesis, or any condition characterized by local or systemic MIF release or synthesis, comprising administering an effective amount of a compound having an isoxazoline moiety, wherein the compound forms an interaction with MIF protein. For example, the compound may bind to MIF protein, thereby mterfermg whh the biological and/or enzymatic activity of MIF protein. The binding may be reversible or irreversibk.
In accordance with the activity of MIF to interfeţe with the anti-mflammatory efîects of skaroids (soch as the anti-mflaiiimatory glucocorticoids), theu»npouDdsofFormidaIorn findtar&erutilitytoenhancethe activity and thetapeuticbeneihsofbothendogcaiousryar
steroidal anti-fflftanmatory ageats. Suchbenefitsmay, insomecases, bemost evident by a reduced need for steroid therapy (e.g., lower dose amount or frequency; less potent agent; reduced need for systemic administration) or by reduced side-effects associated with steroid administration. The benefits of administering a MIF inhibitor (and specifically a compound of Formula I or H) may be realized as a monotherapy, using only the MIF inhibitor of Ihe present invention, or as a combination therapy with additional anti-inflammatory agents, including especially, but without limitation, an anti-inflammatory steroid. Such combination therapy may be achieved through administration of a single formulation or pharmaceutical composition mat combines the MIF inhibitor (particularly an inhibitor of Formula I or n) with at least one other anti-inflammatory agent (which
may be a steroidal or a non-steroidal anti-inflammatory agent), or through administration of separate formulanons or pharmaceutical compositions in conjunction with each other, or both.
Compounds of Formulas I and n are also capable of inhibiting pro-inflammatory cytokines affected by MBF, such as IL-1, IL-2, IL-6, IL-8, IFN-y and TNF, andarethereforeofuseintberapy. IL-1, IL-2, DL-6, IL-8, IFN-y and TNF affect a wide variety of cells and tissues and these cytokines, as well as other leukocyte-derived cytokines, are important and criticai inflammatory mediators of a wide variety of disease states and conditions. The inhibition of these cytokines is of benefit in controUing, reducing and alleviating many of these disease states.
Accordingry, the prestat mvention provides a method of tresting a cytokine mediated disease whtch comprises administering an eflectrve cytokipe-interfering amount of a compound of Formula I or n or a phannaceuticalry acceptable şah thereof.
In particular, compounds of Formulas I or n or a phannaceutically acceptable salt thereof are of use in the therapy of any disease state in a human, or other mammal, which is exacerbated by or caused by excessive or unregulated MDF, IL-1, IL-2, DL-6, IL-8, IFN-y and TNF production by such mammal's cells, such as, but not limited to, monocytes and/or macrophages.
Accordingly, in another aspect, tbis invention relates to a method of inhibiting the production of IL-1 in a mammal in need thereof which comprises administering to said mamţrmţ an effective amount of a compound of Formula I or n a phannaceutically acceptable şah thereof. There are many disease states in which excessive or unregulated IL-1 production is implicated in exacerbating
and/or causing the disease. These include rheumatoid arthritis, osteoarthritis, meningitis, ischemic and hemorrhagic stroke, neurotrauma/closed head injury, stroke, endotoxemia and/or toxic shock syndrome, other acute or chronic inflammatory disease states such as the inflammatory reaction induced by endotoxin or inflammatory bowel disease, tuberculosis, atherosclerosis, muscle degeneration, multiple sclerosis, cachexia, bone resorption, psoriatic arthritis, Reiter's syndrome, rheumatoid arthritis, gout, traumatic arthritis, rubella arthritis and acute synovitis. Recent evidence also links IL-1 activity to diabetes, pancreatic cells disease, and Alzheimer's disease.
In a further aspect, tins inventkm relates to a method of inhibiting the productkm of TNF in a marnmal in need thereof whkh comprises administering to said nMir)fiiTna| an effectivc amount of a compound of Formula I or n or a phannaceuticallyacceptaDk şah thereof. Excessrve or unregulated TNF production has been ţmf'fa'irf'** inmediating or exacerbating a number of diseases including rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and other arthritic conditions, sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, adult respiratory distress syndrome, stroke, cerebral malaria, chronic obstructive puhnonary disease, chronic pulmonary inflammatory disease, silicosis, puhnonary sarcoidosis, bone resorption diseases, such as osteoporosis, cardiac, brain and renal repertusion injury, graft vs. host reaction, allograft rejections, fever and myalgias due to infection, such as influenza (including HIV-induced forms), cerebral malaria, meningitis, ischemic and hemorrhagic stroke, cachexia secondary to infection or malignancy, cachexia secondary to acquked immune deficiency syndrome (AIDS), AIDS, ARC (AIDS
l related complex), keloid formation, scar tissue formation, inflammatory bowel disease, Crohn's disease, ulcerative colitis and pyresis.
Compounds of Formula I or n are also useful in the treatment of viral infections, where such viruses are sensitive to upregulation by TNF or will elicit TNF production in vivo. The viruses contemplated for treatment herein are those that produce TNF as a result of infection, or those which are sensitive to inhibition, such as by decreased replication, directly or indirectly, by the TNF inhibiting-compounds of Formula I or n. Such viruses include, but are not limited to HTV-1 , HTV-2 and HTV-3, Cytomegalovirus (CMV), Influenza, adenovirus and the Herpes group of viruses, such as but not limited to, Herpes Zoster and Herpes Simplex. Accordmgry, in a further aspect, thts invention relates to a method of treating a
Dapiimiiunodefra
administering to such maxnmal an effcctrve TNF mtiMting amount of a compound of Formuk I or H or a pharmaceutkally acceptable şah mereof
Compounds of Formula I or n may also be used in association with the veterinary treatment of mammals, other mân in humans, in need of inhibition of TNF production. TNF mediated diseases for treatment, in animals include disease states such as those noted above, but in particular viral infections. Examples of such viruses include, but are not limited to, lentivirus infections such as, equine infectious anaemia virus, caprine arthritis virus, visna virus, or maedi virus or retrovirus infections, such as but not limited to feline immunodeficiency virus (FIV), bovine immunodeficiency virus, or canine immunodeficiency virus or other retroviral infections.
The compounds of Formula I or n may also be used topically in the
treatment of topica! disease states mediated by or exacerbated by excessive cytokine production, such as by IL-I or TNF respectively, such as infiamed joints, eczema, contact dermatnis psoriasis and other infiammatory skin conditions such as sunburn; infiammatory eye conditions including conjunctivitis; pyresis, pain and other conditions associated with inflammation. Periodontal disease bas also been implemented in cytokine production, both topically and systemically. Hence, the use of compounds of Formula I or n to control the inflammation associated with cytokine production in such perorai diseases such as gingivitis and periodontitis is another aspect of the present invention.
Compounds of Formula I or n have also been shown to inhibit the production of IL(herleukm-«, NAP). Accordingfy, m a further aspect, this invention relates to a method of mhibitmg the productktt of IL-8 in a mammal in need thereof which compriscs ""'""«ffrii1 to said manunal an effective amount of a compound of Formula I or D or aphannaceuticaUy acceptable şah thereof.
There are many disease states in which excessive or unregulated IL-8 production is implicated in exacerbating and/or causing the disease. These diseases are characterized by massive nevrtropbil infiltration such as, psoriasis, infiammatory bowel disease, asthma, cardiac and renal reperfusion injury, adult respiratory distress syndrome, thrombosis and glomerulonephritis. Aii ofthese diseases are associated with increased IL-8 production which is responsible for the chemotaxis of neutrophils into the infiammatory site. In contrast to other inflammatory cytokines (BL-1, TNF, and IL-6), IL-8 has the unique property of promoting neutrophil chemotaxis and activation. Therefore, the inhibition of IL-8 production would lead to a direct reduction hi the neutrophil infiltration.
The compounds of Formula I or II are administered in an amount sufficient to inhibit a cytokine, m particular MIF, IL-1, IL-2, IL-6, DL-8, IPN-y and TNF, production such that h is regulated down to normal levels, or in some case to subnormal levels, so as to ameliorate or prevent Ine disease state. Abnorma! levels of MIF, IL-1, IL-2, EL-6, BL-8, IFN-y and TNF, for instance in the context of the present invention, constitute: (i) levels of free (not cell bound) MIF, IL-1, IL-2, IL-6, IL-8, IFN-Y and TNF greater mân or equal to l picogram per ml; (ii) any cell associated MIF, IL-1, IL-2, IL-6, IL-8, IFN-Y and TNF; or (iii) the presence of MIF, IL-1, IL-2, IL-6, IL-8, IFN-y and TNF mRNA above basal levels in cells or tissues in which MIF, IL-1, IL-2, IL-6, IL-8, IFN-y and TNF, rcspcctively, is produced.
Asusedbcarem,thetatmttinhftHtmgthcp DL-8, IFN-Y «nd TNF" refen to:
a) a deorease of excesnve m vrvo levels of the cytokme MIF, IL-1, IL-2, IL-
6, IL-8, BPN-Y «nd TNF in abumanto normal or sub-normal krvels by inbibition of
the in vivo release of the cytokine by all or select cells, incruding but not limited to
monocytes or macrophages;
b) a down regulation, at the transcription level, of excessive in vivo levels
of the cytokine MIF, IL-1, IL-2, DL-6, IL-8, IFN-y and TNF in a human to normal
or sub-normal levels;
c) a down regulation, at the post-transcription level, of excessive in vivo
levels of the cytokine MIF, IL-1, IL-2, IL-6, IL-8, IFN-Y and TNF in a human to
normal or sub-normal levels;
d) a down regulation, by inbibition of the direct synthesis of the cytokine
M3F, BL-1, IL-2, IL-6, IL-8, IFN-y and TNF as a postranslational event to normal or sub-normal levels; or
e) a down regulation, at the translational level, of excessive in vivo levels of tbe cytokineMIF, IL-1, DL-2, IL-6, IL-8, EFN-y and TNF in ahuman to normal or sub-normal levels.
As used herein, tbe term "MIF mediated disease or disease state" refers to any and all disease states in wbich MIF plays a role, either by production or biological or enzymatic (tautomerase and/or oxidoreductase) activity of MIF itself, or by MIF causing or modnlating another cytokine to be released, such as but not limited to IL-1 ,IL-2, IL-6, IL-8, IFN-y and TNF. A disease state m which, for instance, BL-1 is a major component, and whose production or action, is exacerbated or secreted in response to MIF, would therefbre be considereda disease state mediated by MIF.
As used herein, the term "cytokme" refers to any secreted porypeptide mat affectethefuncţtaroofceibandisamoleciite
includes, but is not limited to, monokines and lymphokines, regardless of which cells produce them. For instance, a monokine is referred to as being produced and secreted by a mononuclear cell, such as a macrophage and/or monocyte. Many other cells however also produce monokines, such as natural killer cells, fibroblasts, basophils, neutrophils, endothelial cells, brain astrocytes, bone marrow stromal cells, epideral keratinocytes and B-lymphocytes. Lymphokines are generally referred to as being produced by lymphocyte cells. Examples of cytokines include, but are not limited to Macrophage Migration Inhibitory Factor (MIF), Interleukin-1 (EL-IX Interleukin-2 (EL-2), Interleukin-6 (EL-6), Interleukin-8
(IL-8), Tumor Necrosis Factor-alpha (TNF-a) and Tumor Necrosis Factor-beta (TNF-p).
As used herein, the tema "cytokine interfering" or "cytokine suppressive amount" refers to an effective amount of a compound of Formula I or H which will căuşe a decrease either in the biologica! activity or the level of the cytokine present in vivo or in vitro, or the in vivo level of the cytokine to normal or sub-normal levels, when given to a patient for the treatment of a disease state which is exacerbated by, or caused by, excessive or unregulated cytokine production.
As used herein, the cytokine referred to in the phrase "inhibition of a cytokine for use in the treatment of a HlV-infected human" is a cytokine which is imph'cated in (a) the initiation and/or mamtenance of T cell activation and/or activated T ceU-mediated HTV gene cxpression and/or replication and/or (b) any cytokme-iDediated disease associated problem such as cachexia or muscle decenefation.
As TNF-P (also known as rymphotoxin) bas close structural homology with TNF-a (also known as cachectin) and since each induces similar biologic responses and binds to the same cellular receptor, both TNF-a and TNF-(3 are inhibited by the compounds of the present invention and thus are herein referred to collectively as "TNF" unless specifically delineated otherwise.
These inhibitor compounds of Formula I or n are of aid in determining the signaling pathways invorvement in inflammatory responses. In particular, a definitive signal transduction pathway can be prescribed to the action of lipopolysaccharide in cytokine production in macrophages. In addition to those diseases already noted herein, treatment of stroke, neurotrauma/CNS head injury,
cardiac, brain and renal reperfusion injury, thrombosis, glomerulonephritis, diabetes and pancreatic cells, multiple sclerosis, muscle degeneration, eczema, psoriasis, sunburn, and conjunctivitis are also included.
It is also recognized that both IL-6 and BL-8 are produced during rhinovirus
i
(HRV) infections and contribuie to the pathogenesis of common cold and exacerbation of asthma associated wfth HRV infection fTurner et al.. (1998), Clin. Mec. Dis., Voi. 26, p. 840; Teren et al. (1997), Am. J. Respir. Crit. Care Med., Voi. 155, p. 1362; Grunbcre et al. (1997), Am. J. Respir. Crit Care Med., Voi. 156, p. 609 and Zhu et al.. J. Clin. Invest (1996), Voi. 97, p 421). It bas also been demonstrated in vitro that infection of pubnonary ephbelial cells wtth HRV resuhs in production of IL-6 and IL-8 (&tea&&sL J- Clin. Invest (1995), Voi. 96, p. 549). Epitbclial cells represent thc primary site of infection of HRV. Therefore, another aspect of the prestat invcation is a method of treatmentto reduce inflammation associated with a rhinovirus Mection, not necessarity a direct effect of the virus itself.
Another aspect of the present invention involves the novei use of these cytokine inhibitors for the treatment of chronic inflammatory or proliferative or angiogenic diseases, which are caused by excessive, or inappropriate angiogenesis. Chronic diseases which have an inappropriate angiogenic component are various ocular neovascularizations, such as diabetic retinopathy and macular degeneration. Other chronic diseases which have an excessive or increased proliferation of vasculature are tumor growth and metastasis, atherosclerosis and certain arthritic conditions. Therefore, cytokine inhibitors will be of utility in the blocking of the angiogenic component of these disease states.
The term "excessive or increased proliferation of vasculature inâppropriate angiogenesis" as med herein includes, but is not limited to, diseases which are characterized by hemangiomas and ocular diseases.
The term "mappropriate angiogenesis" as used herein includes, but is not limited to, diseases which are characterized by vesicle proliferation with accompanying tissue proliferation, such as occurs in cancer, metastasis, arthritis and atherosclerosis.
This invention also encompasses mefhods of treating or preventing disorders that can be treated or prevented by the inhibition of ERK/MAP in a m»m«>i>l preferabiy a human, comprising administering to said mammal an efTecthTamountofacompoundofFonmilalorlL Accordingry, Ine present invention provides a method of treating an ERK/MAP Itinase mediated disease in a mammal in need thereof, preferabiy a taman, which compnses administering to said Mamn>|>r an effcctive amount of a compound of Formula I or n or a pharmaceuticaUy acceptaMe şah thereof.
Preferred ERK/MAP mediated diseases for treatment include, but are not limited to psoriatic arthritis, Reiter's syndrome, rheumatoid arthritis, gout, traumatic arthritis, rubella arthritis and acute synovitis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and other armritic conditions, sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, Alzheimer's disease, stroke, ischemic and hemorrhagic stroke, neurotrauma/closed head injury, asthma, adult respiratory distress syndrome, chronic obstructive puhnonary disease, cerebral malaria, meningitis, chronic pulmonary inflammatory disease, silicosis, pulmonary sarcostosis, bone resorption disease, osteoporosis, restenosis, cardiac

reperfusion injury, brain and renal reperfusion injury, chronic renal failure, thrombosis, gtomerularonephritis, diabetes, diabetic retmopathy, maculat degeneration, graft vs. host reaction, allograft rejection, inflammatory bowel disease, Crohn's disease, ulcerative colitis, neurodegenerative disease, multiple sclerosis, muscle degeneration, diabetic retinopathy, macular degeneration, tumor growth and metastasis, angiogenic disease, rhinovirus infection, perorai disease, such as gingivitis and periodontitis, eczema, contact dennatitis, psoriasis, sunburn, and conjunctivitis.
Ine term "treating", as used herein, refers to reversing, alleviating, inhibitinguprogrtofiwpfeventbgthedisarderoTcorKtitionto term applies, or oue or more symptoons of such dtsorder or comfiDon. The term "treatment", as used herein, refers to the act of treating, as "treating" is defined immediatery above.
This invention also encompasses pnarmaceutical compositions for the treatment of a condttion selected firom tbe gronp consisting of arthritis, psoriatic arthritis, Reiter's syndrome, gout, traumatic arthritis, rubella arthritis and acute synovitis, rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and other arthritic conditions, sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, Alzheimefs disease, stroke, neurotravuna, asthma, adult respiratory distress syndrome, cerebral malaria, chronic pulmonary inflammatory disease, silicosis, pulmonary sarcoidosis, bone resorption disease, osteoporosis, restenosis, cardiac and renal reperfusion injury, thrombosis, glomerularonephritis, diabetes, graft vs. host reaction, allograft rejection, inflammatory bowel disease, Crohris disease, ulcerative colitis, multiple sclerosis,
muscle degeneration, eczema, contact dennatitis, psoriasis, sunbum, or conjunctivitis shock in a tnammal, inchiding a human, comprising an amount of a compound of Formula I or O effective in such treatment and a phannaceutically acceptable carrier.
One embodiment of me present invention provides a method for inactivating enzymatic and biological activity of human MIF comprising contacting the human MIF with a compound, or combination of compounds, that forms a stable interaction with at least one amino acid residue of the human MIF. The invention also relates to inhibiting other cytokines affected by MIF activity including ÎL-1, IL-2, IL-6, EL-8, IFN-y and TNF. The invention encompasses methods of treating or preventing disorders mat can be treated or prevented by the inhibhion of the ERK/MAP patbway in amammal, preierabry a human, comprising administeringto said mammal an effective amount of a compound
As an exampfe of the methods of treatment of the present invention, isoxazoUne-containing compounds of the present invention can be used to treat patients with ARDS (acute respiratory distress syndrome). ARDSisoften considered to be an archetypal clinica! response in which the dynamic balance within the immune response shifts toward excessive inflammation and tissue destruction. MIF is expressed in both type H alveolar cells and infiltrating immune cells. MEF levels in me bronchoalveolar lavage of ARDS patients were found to be significantly elevated when compared to control subjects (Donnelly, eţ a)., Nat. Med., 3,320-323 (1997)). Human MIF enhances both TNFa and IL-8 secretion from ARDS alveolar macrophages (ex vfvo) when compared to control cells. Pre-treatment of these cells with anti-MIF antibodies significantly decreases TNFa and
DL-8 prodnction firom ARDS alveolar cells. Moreover, as discussed above under "Background of tbe Invention," rMEF (recombinant MBF) was foimd to override, in a concentraticm-dependent fashion, glucocorticoid-mediated inhibition of cytokine secretion in ARDS macrophages. These were the first data to indicate that the MIF/glucocorticoid dyad is active in cells that hâd undergone pro-inflammatory activation m vivo during human disease (Donnellv. et al.. Nat. Med., 3,320-323 (l 997). Significantly elevated levels of alveolar MIF were found in those at-risk patients who progressed to ARDS compared to those who did not. MIF likely acts as an important mediator to promote and sustain the pulmonary inflammatory response in ARDS. Its prominent expression in ARDS may explain the fulminant course of this disease and pexhaps wfay glucocorticoid treatment has proven disappointing in estebKshed cascs. Thus, phannaceutkal compositions comprising isoxazoline-containing ccmpounds of &e present invention can be used to treat ARDS patients.
As a further example of the methods of treatment of the present invention, isoxazoline-containing compounds of the present invention can be used to treat patients with rheumatoid arthritis. Synovial fluid obtained from the affected joints of patients with rheumatoid arthritis contain significantly greater levels of MIF than those obtained from patients with osteoartnritis or from normal control subjects (Metz. et al.. Adv. Immunol., 66,197-223 (1997); Leech. et al.. Arthritis Rheum., 41,910-917 (1998); Onodera. et al.. Cytokine, 11,163-167 (1999)). As revealed by immunohistochemical staining methods, infiltrating mononuclear cells within the human arthritic joint are the primary source of MIF. In two animal models of arthritis, neutralizing anti-MEF mAb's significantly inhibited disease progression
anddisease severity rLceck et al.. Arthritis Rheum., 41, 910-917 (1998); Mikulowska. et al.. J. Immunol., 158, 5514-5517 (1997)) giving impetus to the desirability of developing additional MIF inhibitors for potenţial therapeutic use in inflammatory disease. Thus, pharmaceutical compositions comprising isoxazoline compounds or isoxazoline-related compounds of the present invention can be used to treat arthritis patients.
In yet a further example of the methods of treatment of the present invention, isoxazoline-containing compounds of the present invention can be used to treat patients with atopic dennatitis. Atopic dermatitis is a chronic pruritic inflammatory sicin disorder. Its palhogenesis, in part, is tix>ught to be due to
DdDdxmty In lesions from
patieats wfth atopic dermatitis, MIF proteîn is diffiBely disttibuted throughout the entireepidennal foyer wnincreasedcaqpressw
FEBSLett, 38 1, 199-202(19%)). fa nora huinan skin, Mff nas primarily been localized to epidemia! ketktmocytes. The scrum MEF level of atopic dermatitis patients were 6-fold higher tium in control subjects. Additionally, serum MIF levels in atopic dermatitis patients decreased as clinica! features improved, suggesting that MIF plays a pivotai role in the inflammatory response in the skin during dermatitis. Thus, pharmaceutical compositions comprising isoxazoline-containing compounds of the present invention can be used to treat patients with atopic dermatitis.
In a similar manner, the present invention also provides a method for treating or preventing other inflammatory or autoimmune disorders including, but not limited to, proliferative vascular disease, cytokine-mediated toxicity, sepsis,
septic shock, psoriasis, interleultin-2 toxicity, asthma, MDF-mediated conditions, insulin-dependent diabetes, multiple sclerosis, grafi versus host disease, lupus syndromes, and other conditions characterized by local or systemic MIF release or
synthesis or by other cytokines affected by MIF. In yet another example of tbe methods of treatment of the present invention, compounds of the present invention can be used to treat patients wfth tumor growth. Neutralizing anti-MIF antibodies have been found to significantly reduce growth and vascularization (angiogenesis) of mouse 38C13 B cell rymphoma in vfvo rChesnev. et ai.. Mol. Med., 5,181-191 (1999)). MIF was expressed predominantly in tumor-associated neovasculature. Cuhured microvascular arîo&eiialccns,biiti>ot38C13Bcdls,wereo
require tfc arrivfty fer proliferai™ m *itm (T»V rrt >| Mol. Med., 4, 707-
714(1998)). Inadditkmthetdministratkmofanti-MffandbodMStom found to significanth/ inhibit tfie aeovascnlnrization reqxxtse elicited by Matrigel implantation, a model of newblood vessel formation in vrvo (Bozza. et al.. J. Exp. Med., 189,341-346 (1999)). These data indicate that MIF plays an important role in tumor angiogenesis, a new target for the development of anti-neoplastic agents that inhibit tumor neovascularization.
Thus, the present invention also provides a method for treating or preventing tumor growth or angiogenesis, comprising administering an effective amount of a compound, or combination of compounds, having an isoxazoline moiety and that forms a stable interaction with at least one amino acid residue of an MEF protein.
The present invention also provides a compound of Formula I or E, or a
phannaceutically acceptable şah mereof, as a pharmaceutical composition comprising either of the aforesaid, for use in a medicine or for the manufacture of a medicament for the treatment or prevention of infiammatory disorders including arthritis, proliferative vascular disease, ARDS, cytokine-fflediated toxicity, sepsis, septic shock, psoriasis, interleukin-2 toxicity, asthma, MIF-mediated conditions, autoimmune disorders (including, but not limtted to, rheumatoid arthritis, insulin-dependent diabetes, multiple scletosis, graft versus host disease, lupus syndromes), tumor growtib or angiogenesis, or any condition characterized by local or systemic MIF release or synthesis.
This invention also encompasses pharmaceutical compositions for the Ireatment of a condition which cao be treated by the mhibrtion of the ERK7MAP kinase pathway in a ™«ninalt in**1" a human, comprising an amount of a compound of Formula I or D eflective in such treatment and a pharmaccuticaUy acceptable carrier.
This invention also encompasses prodrugs of compounds of the Formula I or II and pharmaceutical compositions containing these prodrugs. Compounds of Formula I or n having free amino, amido, hydroxy or carboxylic groups can be converted into prodrugs. Prodrugs include compounds wherein an amino acid residue, or a polypeptide chain of two or more (e.g., two, three or four) amino acid residues which are covalently joined through peptide bonds to free amino, hydroxy or carboxylic acid groups of compounds of Formula I or H. The amino acid residues include the 20 naturally occurring amino acids commonly designated by three letter symbols and also include, 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid,
, uwiuuty»ieuje, homoserine, ornithine and methionine sulfone. Prodrugs also include compounds wherein carbonates, carbamates, amides and alkyl esters which are covalently bonded to the above substituents of formula I through the carbonyl carbon prodrug sidechain. The invention also encompasses sustained release compositions.
One of ordinary skill in tbe art will appreciate tfaat the compounds of the invention are userul in treattng a diverse array of diseases. One of ordinary skill in the art will also appreciate mat when using the compounds of the invention in the treatment of a specific disease mat the compounds of the invention may be combined with various existing menpeutic agents used for mat disease.
For the treatment ofrhrnmatotd arthritis, the compounds of the invention may be combined with agents such as TNF mhibitors such as anti-TNF monoclonal antibodies and TNF receptor immunoglobuhn molecules, COX-2 mhibitors, such as celecoxib, rofecoxib, valdecoxib and etoricoxib, low dose methotrexate, lefunomide, hydroxychloroqume, d-penicillamine, auranofin or parenteral or oral gold.
The compounds of the invention can also be used in combination with existing merapeutic agents for the treatment of osteoarthritis. Suitable agents to be used in combination include standard non-steroidal anti-inflammatory agents such as piroxicam, diclofenac, propionic acids such as naproxen, flubiprofen, fenoprofen, ketoprofen and ibuprofen, fenamates such as mefenamic acid, indomethacin, sulindac, apazone, pyrazolones such as phenylbutazone, salicylates such as aspirin, COX-2 inhibitors such as celecoxib, valdecoxib, rofecoxib and etoricoxib, analgesics and intraarticular therapies such as corticosteroids and
nyaiuronic acios suen as nyaigan and synvisc.
The compounds of the present invention may also be used in combination with aoticancer agents such as endostatin and angiostatin or cytotoxic drugs such as adriamycin, daunomycin, cis-platinum, etoposide, taxol, taxotere and alkaloids, such as vincristine, farnesyl transferase inhibitors, VegF inhibitors, and antimetabolites such as methotrexate.
The compounds of the invention may also be used in combination with antiviral agents such as Viracept, ACT, aciclovir and famciclovir, and antisepsis compounds such as Valant
The compounds of the present invention may also be used in combination
âumch
such as statins, fibrates, beta-biockent, Ace inhibitors, Angiotensin-2 receptor antagoniste and platelet aggtegation miubitors.
The compounds of the present invention may also be used in combination with osteoporosis agents such as roloxifene, droloxifene, lasofoxifene or fosomax and immunosuppressant agents such as FK-506 and rapamycin.
The compounds of the present invention may also be used in combination with CHS agents such as antidepressants, such as sertraline, anti-Parkinsonian drugs such as deprenyl, L-dopa, Requip, Mirapex, MAOB inhibitors such as selegine and rasagiline, comP inhibitors such as Tasmar, A-2 inhibitors, dopamine reuptake inhibitors, NMDA antagonists, Nicotine agonists, Dopamine agonists and inhibitors of neuronal nitric oxide synthase, and anti-Alzheimer's drugs such as donepezil, tacrine, COX-2 inhibitors, propentofylline or metryfonate.
This invention also encompasses pharmaceutical compositions for the
treaiment ot a condinon wtuch can be treated by the inhibition of ERK/MAP kinase in a mammal, including a human, comprising an amonnt of a compound of Formula I or n effective in such treatment and a pharmaceutically acceptable carrier.
The present inventkm further provides a method for treating inflammatory disorders including, but not limited to, arthritis, proliferative vascular disease, ARDS (acute respiratory distress syndrome), cytokine-mediated toxicity, sepsis, septic shock, psoriasis, interleukin-2 toxicity, asthma, MIF-mediated conditions, autoimmune disorders (including, but not limited to, rheumatoid arthritis, insulin-dependent diabetes, multiple sderosis, graft versus host disease, lupus syndromes), tumor growth or angiogenesis, or any condhion dttracterized by local or systemic MBP release or synthesis, comprising admimsteringaneffectrveamountofa compound having an isoxazoKnr moicty, wfaerein tine isoxazoline moiety forms a stable covalent intenctkm wfth at kast one amino acid residue of an MIF protein. Preferabry, the interactkm occurs at or near tf»e active site of the tautomease actrvity of the MEF protein. The present invention also provides a pharmaceutical composition comprising a compound having an isoxazoline or isoxazoline-related moiety and a phannaceutically acceptable carrier, wherein the moiety forms a stable covalent interaction with at least one amino acid residue of a MIF protein.
The present invention relates to compounds, compositions, processes of making, and methods of use related to inhibiting Macrophage Migration Inhibitory Factor (MBF) activity. The compounds comprise a genus of low molecular weight compounds comprising optionalry substituted isoxazoline ring systems that act as inhibitors of MIF, and also inhibiting other cytokines affected by MIF activity
including BL-1, D.-2, tt,-6, EL-8, IFN-y and TNF. This invention. also encompasses methods of treating or preventing disorders that can be treated or prevented by the inhibition of tibe ERK/MAP pathway in a mammal, preferably a human, comprising administering to said mammal an effective amount of a compound. The compounds are useful for treating a variety of diseases involving any disease state in a human, or other mammal, which is exacerbated by or caused by excessive or unregulated MF, IL-1, 1L-2, IL-6, H8, IFN-y and TNF production by such mammal's cells, such as, but not limfted to, monocytes and/or macrophages, or any disease state mat can be modutted by inhibiting the ERK/MAP pathway.
One embodiment of the invention provides a new class of MDF and other cytokine tnbibitors structuratty related to isoxazoline which are suitable to tutrah bou endogenois and entogenous MIF and other cytokines. Thepresent invention merefore provides a genus of inhibitor compounds. Compounds in mis genus are generaUy described by tibe general Formulas I and n herein. Unless otherwise indicated, structural Formulas I and n and described substituents are as indicated herein.
Given the teachings herein, me compounds can be synthesized by a variety of routes known to the organic chemist having ordinary skill in the art.
EXAMPLES
Having generaUy described tbis invention, a further understanding can be obtained by reference to certain specific examples, which are provîded herein for purposes of illustration only and are not intended to be limiting unless otherwise specified.
EXAMPLE1
(Figure Removed)

Referring now to the Phenyl Serie* reaction scheme in FIG. IA:
To the sohjtkm of Chlorooximc (Compound 8,14.8 g) in THF (100 ml) was added triethylamine (14.2 g) and Ac sohition was cooled to 5 -10 deg. To the above solution was added slowiy methylstyryl acetate (S g) and the resuttant solution was stirred at RT for 24 hrs. The solvent was removed by distillation and the residue was dissolved in ethyl acetate (lOOml) and washed with water (2x5Oml) foliowed by brine solutioa The organic layer was dried over anhydrous sodium sulfate, fihered, and concentrated to a residue. The TLC shows that two regioisomers were formed (Compounds 23a and 23b). The yellow solid which was a mixture of the two regioisomers (25 g) was taken on to the hydrolysis step. The crude reaction mass (Compounds 23a and 23b, 25 g) was taken in
methanol (200ml) and 25% sodium hydroxide solution (l 3.0 ml) was added. The resuhant solution was refluxed for 2 hrs. The solvent was removed by distillation and the residue was diluted with water (lOOml) and adjusted to a pH of 2 with hydrochloric acid (2M). The compound was extracted with ethyl acetate (2 x 200ml). The organic layer was furtber washed with brine (lOOml). The resultant organic layer was dried over anhydrous sodium sulfate, fîltered, and concentrated. The mixture of the two isomers (Compounds 24a and 24b) was purified by column chromatography (100-200 mesh silica gel, 50% Ethyl acetate - Petether ) to give 24a (0.700 g) as a white solid. The material was taken on without further characterization.
The benzylated acid derivativ« HPLC Condiţiona:
Column : SymmetryshieldRP-18 (4.6xl50)mm
Max: Mobile phase : 0.01M KH2P04 (PH = 2.5) : Acetonitrile (70:30)
Flowrate : l.OmL/min ; Wavelength : 215nm
Retentiontime : 13.33 ; Purity : 92.38%
ER( KBr, v max) : 3382,3035,1704,1607,1516,1433,1219,1173, 872.
1H NMR: (DMS(Xd6,300 MHz); 6 12.3(br.s, 1H), 9.9(br.s, 1H), 7.3(d,2H), 7.2(m,5H), 6.8(d£H), 4.8{d,lH), 4.7(m, 1H), 2.7(m, 2H). Mam:m/z. 298(M+1).
To the benzyiated acid derivative (Compound 24a, 0.300 g) was added thionyl chloride (l ml) at O deg and the rcsuhant clear solution was stirred at RT for 30 min. The excess thionyl chloride was removed under reduced pressure (lOmm-Hg). To the residue was added isobutyi alcohol (1.6 g) and the reaction mixture was stirred at RT for 2 hrs. The solution was diluted with ethyl acetate (5ml) and washed with water (2x5ml). The organic layer was concentrated to a residue. The Compound 26a was purified by cohonn chromatography (60-120 mesh silica gel, 20%Etlrylacetate-Petcther)toyieW26a(0.150g)»saliquid.
The benzylated acid derivative (Compound 26*, 0.150 g) was dissorved in ethanol (15 mi) and 10% pafladhim on carbon was added. The reaction mixture was hydrogenated using balloon pressure for 4 hrs. The reaction mixture was filteredthroughapedofcxmt:aiKlbedwaswashedwithrK>tetharK)l(30mlX The ethanol was evaporated to give a residue. The ester 27a was further purified by column chromatography using 100-200 mesh silica gel and 40% Ethyl acetate -Petether as eluent to yield 27a (0.060 g) as a solid. M.P : 143 - 146 deg. HPLC Condiţiona:
Column : ZorbaxSBC-18 (4.6x250) mm
Max: Mobile phase : 0.1 % TFA : Acetonitrile (50:50)
Flowrate : l.OmL/min ; Wavelength : 275 nm
Retention time : 18.93 min ; Purity : 95.10%
IR ( KBr, v max) : 3407,2961,1707,1608,1516,1428,1346,1279,1213,
1169,1050,1005, 877,700 cm-1. 1H NMR : (DMSO-d6,300 MHz); 6 9.8(s, 1H), 7.6(dH), 7.3-7.5(m, 5H), 6.8(d, 2H), 4.9(d, 1H), 4.8(m, 1H), 3.9(d, 2H), 2.8(2dd, 2H), 1.9(m, 1H), 0.9(d, 6H). Mass: m/z. 354(M+1), 235;
To the benzylated acid derivative (Compound 24a, 0.300 g) was added thionyl chloride (Iml) at O deg and the resuhant clear solution was stirred at RT for 30 min. The excess thionyl chloride was removed under reduced pressure (lOmm-Hg) and to the residue was added isobutyl amine (l .46 g) and the reaction stirred at RT for 2 hrs. The solution was dihited with ethyl acetate (5ml), washed with water (2x5ml) and the organic layer was concentrated to a residue. The compound 28a was purifîed by colmnn chromatography (60-120 mesh silica gel, 30% Ethyl acetate - Petether) to give pure 28a (OJ80 g) as a Kqukl. The compound was taken on to the next step wrthout rbrther characterization.
The benzylated amide derivative (Compound28a,0.150g) wasdissolved in ethanol (15 ml) and 10% palladram on carbon was added. The reaction mixture was hydrogenated nsmg balloon pressure fot fourhours. The reaction mbrture was fîltered through a pad of ceKte and the bed was washed with hoţ ethanol (30ml). The ethanol was evaporated to gtve a residue. The compound 29a was further purified by column chromatography using 100-200 mesh silica gel and 40% Ethyl acetate - Petether as eluent to give the desired product 29a as a solid (0.060g). M.P:144-149°C. HPLC Conditions:
Column : SymmetryC-18 (4.6x250) mm
Max: Mobile phase : 0.01 M KH2PO4 (PH = 2.5) : Acetonitrile (60 :40)
Flowrate : 0.6mL/min ; Wavelength : 270 nm
Retention time :21.38min; Purity : 92.35%
ER ( KBr, v max): 3373,2959,1647,1607,1517,1440,1273,1171,882, 839,700 cm-1. 1H NMR: (CDC13,300 MHz), 8 7.5(d,.2H), 7.3(m, 5H), 6.8(d, 2H), 6.1-6.2(2br.s, 2H), 4.8(m, 1H), 4.7(d, 1H), 3.1(m, 2H), 2.7(m, 2H), 1.8(m, 1H), 0.8(m, 6H). M«ss:ni/z.353(M-H),335,232.
Referring nowto the Phenyl Series reactkm scheme in FIG. 1B: To the sohition of Chlorooxime (Compound g, 14.8g) in THF (100 ml) was added triethylamine (14.2 g) aad tbe solution was cooled to 5 -10 deg. To the above solution was added slowfry Methvlstyryl acetate (5.0 g) and the resultant sohition was stirredatRT for 24 hrs. The solvent was thcnremoved by distillation and Ac residue was dissolved in ethyl acetate (lOOml) and washedwi&water (2x50ml) fbllowed by brine solutkm. The organic kyer was dried over anbydrous sodtum suh%te aad concenteated to a lesâdue. The TLC shows that two regtoisomers wece fooned (Compcmnds 23* and 23b). The yellowish solid mixture of the two regioisomers (aude mass 25 g) was taken into the hydrolysis step,
The crude 23a and 23b (25.0g) were taken in methanol (200ml) and sodium hydroxide solution (25%, 3.24g) was added and the resultant solution was refluxed for 2 hrs. The solvent was removed by distillation and the residue was diluted with water (lOOml) and acidified to a PH of 2 with hydrochloric acid (2M). The compound was extracted with ethyl acetate (2 x 200ml). The organic layer was further washed with brine (lOOml). The resultant organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The mixture of two isomers (Compounds 24a and 24b) was ftirflier purifîed by cohimn chromatography (100-200 mesh silica gel, 50% Ethyl acetate - Petether) to yield 24b (1.1 g) as a
white crystalline solid The compound was taken on to the next step.
The benzyiated acid derivative(24b, 0.300g) was dissolved in ethanol (60ml) following which 10% Palladium on carbon (0.060g) was added. The reaction mixture was hydrogenated using balloon pressure for four hours. The reaction mixture was fîltered over through a bed of Celite and the bed was washed with ethanol(30mJ). The ethanol was evaporated to give 25b as a residue. The compound 25b was further purified by column chromatography using 60-120 mesh silica gel and 10% Methanol - Chloroform as eluent to yield 25b as on off white solid (O.OSOg) (m.p. 153 -159-C).
IR ( KBr, v max) : 3150,1707,1600,1517,1437,1350,1275,961,755 cm-1. 1H NMR: (CD,OD, 300 MHz); S 73(d, 2H), 7.2(m, 5H), 6.8(d, 2H), 5.5(d, 1H), 4.0(m, 1HX 2.7(m, 2H). Mân : mfe. 296(M-1X 252,171,133.
To the benzyiated acid derivative (24b, 0.300g) was added thkmyl chloride (Iml)at0deg. TheresuftantckarsohjtkmwasstiiTedatRTfo excess of thionyl chloride was removed under reduced pressure (lOnxm-Hg). To the residue was added isobutyl alcohol (1.6g) and the solution stirred at RT for 2 hrs. The solution was diluted with elhyl acetate (5ml) and extracted with water (2x5ml). The organic layer was concentrated to a residue. The Compound 26b was purified by column chromatography (60-120 mesh silica gel, 20% Elhyl acetate - Pet.ether). The product was isolate (180 mg) as a liquid.
The benzyiated acid derivative (Compound 26b, 0.150 g) was dissolved in ethanol (15 ml) and Palladium on carbon (0.030 g) was added. The reaction mixture was hydrogenated using balloon pressure for 4 hrs. The reaction mixture was filtered through a pad of celite and the bed was washed with hoţ ethanol (30
ml). The ethanol was cvaporated to give a residue. The compound 27b was further
purified by cotumn chromatography using 100-200 mesh silica gel and 40% Bthyl
acetate - Petether as eluent to give the desiied 27b as an off-white solid (80 mg).
M.P.:136-143C.
HPLC Conditkms:
Cohimn : SymmetryshieldRP-18(4.6xl50)mm
Mobile phase : 0.01M KH2PO4 (PH = 2.5): Acetonitrile (40:60)
Flowrate : l.OmL/min.; Wavelength : 275 nm
Retention time : 7.13 min.; Purity : %.4%
IR ( KBr, v max) : 3174,2965,1735,1601,1517,1350,1274,1171,752 cm-1.
1H NMR: (CD30D, 300 MHz); 8 7.6(dHX 73-7(m, 5H), 6.8(d, 2H), 53(d,
1H), 4.1(m, 1H), 3.9(m, 2H), 2.8(2dd, 2H), 1.9(m, 1H), 0.9(d, 6H). MMI : m/z.
354(M+1), 335,307.
To the benzylated acid derivative (Compound 24b, 0300 g) was added thionyl chloride(l ml) at O deg and Ae resuhant ctear solution was stirred at RT for 30 min. The excess of thionyl chloride was removed under reduced pressure (lOmm-Hg). To Ae residue was added isobutyl amine (2 ml) and the solution was stirred at RT for 2 hrs. The solution was then diluted with ethyl acetate (5ml) and washed with water (2x5ml). The organic layer was concentrated in vacuo to a residue. The compound 28b was purified by column chromatography (60-120 mesh silica gel, 30% Ethyl acetate - Petether) to give 28b (170 mg) as a liquid
To a solution of benzylated amide derivative (Compound 28b, 0.150 g)) in ethanol (l 5 ml) was added Palladium on carbon and the reaction mixture was hydrogenated using balloon pressure at rt for four bour s. The reaction mixture was
filtered througb a bed of cetite and the bed was washed with hoţ ethanol (30ml).
The ethanol was evaporated to grve a residue. The compound 29b was further
purified by column chromatography using 100-200 mesh silica gel and 40% Ethyl
acetate - Petether as eluent to give 0.060g of pure 29b as a solid. MJP : 185 -
190°C.
HDPLC Condiţiona:
Column : Symmetry C-18 (4.6 x250) mm
Mobile phase : 0.05% TFA: Acetonitrile (55 : 45)
Flowrate : l.OmL/min.; Wavelength : 210 nm
Retention time : 13.57 min.; Purity : 98.16%
IR ( KBr, v max) : 3396,
2%l,1649,1606>1544>1441,13461278,1241,n72,839,747cm-l.
1H NMR: (DMSOd6,300 MHzX 8 8.1(t, IHX (»*•«, IHX 7. 2HX 73(m, 5H),
6.8(d, 2H), 5.X4 IHX *.0(in, IHX 3.0(m, 2H), 2.5(m, 2HX 1.7(m, 1H), 0.8(m,
6H). Mass: m/z. 353(K*HX 335» 234.
EX AMPLE 2
(Figure Removed)

ReferringnowtolfaePropyl Series reaction scheme in FIG. 2A: To the sohrtion of4-Hydioatybcpaaldetyde (Compound 5,10 g) in THF (200 ml), was added potassiuni carbonate (16.95 g) followed by benzyl bromide (16.8 g) and the tentant nsaction mixture was refruxed for 24 hrs. The leactkm mixture was cooled toRT and THF was removed under reduced pressure (lOmm-Hg). The residue was dissolved in ethyl acetate (100 ml) and washed with water (100 ml) followed by brine (100 ml). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Evaporation of the solvent gave residue which was triturated with petether to give a crystalline solid. The solid compound was filtered, washed with petether, and dried under reduced pressure(10mm-Hg) to give an off-white crystalline solid (Compound 6,15.6 g). To the solution of benzyiated derivative (Compound 6,10 g) in methanol (100 ml) was added hydroxylamine hydrochloride (4.9 g) and sodium acetate (9.6 g). The resultant reaction mixture was refluxed for 3 hrs. The reaction mass was cooled to RT. The solvent was removed under reduced pressure (lOrnm-Hg), the
residue was dissolved in ethyl acetate (lOOml), and washed with water (lOOml) followed by brine (lOOml). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Evaporation of the solvent gave a white crystalline sotid which was rinsed with pet.ether and dried under reduced pressure (lOmm-Hg) to give compound 7 (8.0 g).
To the solution of Oxime derivative (Compound 7, l O g) in THF (l 00 ml) was added N-chlorosuccinimide (8.8 g) in THF at O deg over a period of 30 minutes and the resultant solution was stirred at O to 5 deg for 2- 3 hrs. The solvent was evaporated at 40 deg under reduced pressure. The residue was dissolved in ethyl acetate (lOOml) and washed with water (lOOml) followed by brine (lOOml). The organic layer was dried over anhydrous sodium sulfate, fihered, and concentrated to a residue. The residue was washed with hexane and dried under reduced pressure (lOmm-Hg) to give a h'ght yellow solid (Compound 8,11.0 g).
To the solution of Chlorooximc (Compound 8,16.74 g) in THF (100 ml)
was added trkthylanrâe (l 4.2 g) aml the reactiv was coolcd to 5 -10
deg. To this solution was added slowry methyl-3-heptenoate (4.5 g) and the resultant solution was stirred at RT for 24 hrs. The solvent was removed under reduced pressure (lOmm-Hg) and the residue was dissolved in ethyl acetate (lOOml), washed with water (2x50ml) followed by a brine solution. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to a residue. The TLC shows mat two regioisomers were formed (Structure 9a and Structure 9b). The crude mass of the two regioisomers (25 g) was taken on the for hydrolysis step.
The crude reaction mass (25 g) was taken in methanol (200ml) and added
sodium hydroxide sohjtion(25%, 13.6ml). The resuitant solution was refluxed for 2 bis. The solvent was removed by distillation and the residue was diluted with water (lOOml) and the pH was adjusted to 2 with hydrochloric acid (2M). The compound was extracted with ethyl acetate (2 x 200ml). The combined organic layers was again washed with brine (lOOml) and the resuitant organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture of two isomers was further purified by column chromatography (100-200 mesh silica gel, 50%.Ethyl acetate - Petetber) to give compound lOa (0.700 g) as a white solid.
The Acid derivative (Compound lOa, 0.300 g) was dissolved in ethanol (30 ml) and palladium on carbon (0.030 g) was added. The solution was hydrogenated usmgbaUoonprcssurefor 4hours.Thereactionmixture was filtered over apadof cetite and the bed was washed whh ethanol (30ml). The ethanol was evaporatedto give a residue. The product was further purified by column cbromalography using 60-120 mesh silica gel and 30% Ethyl acetate -Petcther as etuent to give l la (0.060 g) as an off-whtte solid. MJP: 175 - 177 C HPLC Condiţiona:
Column : SymmetryshieldRP-18 (4.6x150) nm
Max: Mobile phase : 0.01 M KH2PO4 (PH = 2.5): Acetonitrile (65:35)
Flowrate : l.Oml/min ; Wavelength : 210 nm
Retentionume : 5.49 min. ; Purity : 94.44%
ER (KBr, vmax): 3372,3284,2931,1703,1607,1516,1435,1351,1283,1220, 1171,943,878,834,674 cm-1. 1H NMR: (DMSO-d6, 300 MHz) ; 8 12.2(br.s, 1H), 10(br.s, 1H), 7.5(d, 2H), 6.8(d, 2H), 4.7(m, 1H), 3.5(m, 1H), 2.5(m, 2H), 1.2-1.4(m, 4H), 0.8(t, 3H). Mân: m/z, 263(M+1) , 219,178.
To tbe Compound lOa (0300 g) at O deg was added thionyi chloride (îmi) and the resuftant clear sohition stirred at RT for 30 min. The excess thionyi chloride was removed under reduced pressure (lOmm-Hg) and to tbe residue was added isobutyl alcohol (1.6 g) and the resultant solution was stirred at RT for 2 hrs. The sohition was diluted with ethyl acetate (5 ml) and washed with water (2x5ml). The organic layer was concentrated to a residue which was purifîed by column chromatography (60-120 mesh silica gel, 20% Ethyl acetate- Petether) to give 12a (0.150g)asaliquid.
The compound 12a (0.150 g) was dissolved hi ethanol (15 ml) and 10% palladium on carbon (0.30 g) was added. The sohition was hydrogenated using belloon pressure for 4 hrs. Theieactkmmixture wasfiheredoverapadof colite and the bed was washed wifli bot ethanol (30 ml). The ethanol was evaporatedto give a residue which was further porified by cohnnn chromatDgraphy using 100-sihca gel and 30% Ethyl «cetate- Pet eiher as eh to give 1 3a (0.060
g) as a h'quid. Yield : 60mg . HPLC Condiţiona :
Column : SymmetrySheildRP-18(4.6xl50)
Max: Mobile phase : 0.01 M KH2PO4 (PH = 5) : Acetonitrile
Flowrate : 1.0 ml /min ; Wavelength : 270 nm
Retention time : 5.82 min ; Purity : 96.30%
IR(KBr, vmax) : 3390, 2961, 1729, 1607, 1516, 1464, 1350, 1272, 1173, 738 cm-1. 1H NMR : (CDC13, 300 MHz); 8 7.5(d, 2H), 6.9(d, 2H), 4.8(m, 1H), 3.9(d, 2H), 3.4(m, 1H), 2.6(2dd, 2H), 1.9(m, 1H), 1.3-1.5(m, 4H), 0.8(m, 9H). Mass : m/z . 320(M+1).
To compound lOa (0.300 g) was added thionyl chloride (l ml) at O deg and the resuhant clear solution stirred at RT for 30 min. The excess of thionyl chloride was removed under reduced pressure (lOmm-Hg) and to the residue was added isobutyl amine (l .46 g) and the solutions was stirred at RT for 2 hrs. The solution was dihited with ethyl acetate (5 ml) and extracted with water (2x5ml). The organic layer was concentrated to a residue which was purified by column ehromatography (60 -120 mesh silica gel, 30% Ethyl acetate- Petether) to give 14a (0.180 g) as a liquid.
The compound 14a (0.150 g) was dissolved in ethanol (15 ml) then 10% palladium on carbon (0.030 g) was added. The sohition was hydrogenated using balloon pressure for four hours. The leactkm mixture was firtered through a pad of celhe and the bed was washed wfth bot ethanol (30ml). The ethanol was evaporated to give a residue which was turther purified by cohimn ehromatography using 100-200 mesh silica gel and 30% Eflryl acetate- Petether as ehient to give 15a (0.060 g) as a solid. M.P: 136.6 -1435 deg. HPLC Condiţiona:
Column : ZorbaxSBC-18 (4.6x250) mm
Max: Mobile phase : Water: Acetonitrile (60:40)
Flowrate : l.Oml/min ; Wavelength : 220 nm
Retention time : 10.28 min ; Purity : 95.10%
IR ( KBR, v max) : 3296,2934,1646,1608,1517,1462,1352,1278,1172, 880, 838,605 cm-1. 1H NMR: (CDC13,300MHz ); 6 7.5(d, 2H), 6.8(d, 2H), 6.3(br.t, 1H), 4.8(m, 1H), 3.4(m, 1H), 3.1(m, 2H), 2.6(2dd, 2H), 1.8(m, 1H), 1.3-1.5(m, 4H), 0.8(m, 9H). Mass: m/z: 319(M+1), 301,200.
Referring now to the Propyl Series reaction scheme in FIG. 2B:
To the solution of 4-Hydroxybenzaldehyde (Compound 5,10.0 g) in THF (200 ml), was added potassium carbonate (l 6.95 g) followed by benzyl bromide (16.8 g) and the resultant reaction mixture was refluxed for 24 hrs. The reaction mixture was cooled to RT and the THF was removed under reduced pressure (lOmm-Hg). The residue was dissolved in ethyl acetate (lOOml) and washed with water (lOOml) followed by brine (lOOml). The organic layer was dried over anhydrous sodium sulfate. Evaporation of the solvent gave residue. The residue was decanted with petether gave crystalline solid. The solid compound was fihered and washed wim petether and dried under reduced pressure(10mm-Hg) to giveanoff-whHea75tal]inesolid(Coiiapouod6,15.6g), The compound was taken on without turther chaiacteiization.
To the sohrtk» of bcnzylatwlderivatwc (Compound 6,10.0 g) m methanol (100 ml) was added hydroxylamine hydcochloride (4.9 g) and sodium acetate (9.6 g). The resultant reaction mixture was refluxed fot 3 hrs. The reaction mass was cooled to RT, the solvent was removed under reduced pressure (lOmm-Hg), and the residue was dissolved in ethyl acetate (lOOml) and washed with water (lOOml) followed by brine (lOOml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give a white crystaUine solid (compound 7), which was rinsed with petether and dried under reduced pressure (lOmm-Hg) to give 8.0g of 7. The compound was taken on without further characterization.
To the solution of Oxime derivative (Compound 7,10.0 g) in THF (90 ml) was added Nhlorosiiccmimioe (8.8 g) in THF (10 ml) at O deg over a period of 30 minutes and the resultant solution was stirred at O to 5 deg for 2- 3 hrs. The solvent
was evaporated at 40 deg under reduced pressure. The residue was dissolved in ethyl acetate (lOQml) and washed whh water (lOOml) foUowed by brine (lOOml). The organic layer was dried over anhydroas sodium sulfate, filtered, and concentrated to a residue. The residue was washed whh hexane to give a crystalline solid (Compound 8) which upon drying under reduced pressure (lOmm-Hg) gave 11.0 g of chloro-oxime 8 as a light yellow seini solid. The product was taken on whhout further characterization.
To the solution of Chlorooxime (Compound 8,16.74 g) in THF (100 ml) was added triethyiamine (14.2 g) and the reaction mixture was cooled to 5 -10 deg. To this sonition methyl-3-heptenoate (4.5 g) was slowry added and the resahantsohitkmwasstmedatRTfor24hrs.Thesofrert reduced pressure (lOmm-Hg). The residue was dissolved in ethyl acetate (lOOml) and washed with water (2x50ml) and brine sotunon. The organic laycr was dried over anhydrous sodium sulfate, filtered, and concentrated to a residue. The TLC shows mat two regjoisomers were formed (Structure 9a and Structure 9b). The yellow solid crude mass (25 g) of the two regioisomers was taken on to the hydrolysis step. TLC System t 20% Ethyl acetate - Petether. Rf: 0.4
The crude reaction mixture of 9a and 9b (25g) was taken up in methanol (200ml) and sodium hydroxide solution (25%, 13.6ml) was added. The resultant solution was refluxed for 2 hrs. The solvent was removed by distillation and the residue was diluted with water (l OOml) and the pH adjusted to 2 with hydrochloric acid (2M). The solution was extracted with ethyl acetate (2 x 200ml) and the combined organic layers was again washed with brine (lOOml). The resultant
organic foyer was dned over anhydrous sodium sulfate, filtered, and concentrated. The mixture of the two isomers (Compounds lOa and lOb) was further purified by column chromatography (100-200 mesh silica gel, 50%.Ethyl acetate - Petether) to give l Ob (1.3 g) as a wbite crystaUine solid which was taken on without further characterization.
To the Compound lOb (0.300 g) at O deg was added thionyl chloride (l ml) and the resultant clear solution stirred at RT for 30 min. Excess thionyl chloride was removed under reduced pressure (lOmm-Hg) and isobutyl alcohol (l .6 g) was added to the residue and the solution was stirred at RT for 2 hrs. The solution was dihited with ethyl acetate (5mT) and washed wfth water (2x5ml). The organic layer was concentrated Io a residue and compound 12b was purified by cohimn chromatograpby (60-120 mesh afica gel, 20% Eftţyl acetate- Petether) to give 12b (0.150g)«s»hqoid The material was taken ontothencxt step.
Palladhmn on carbon (0.030 g) was added The sohition was hydrogenated using
belloon pressure for 4 hrs. The solution was fihered through a pad of celite and the
bed was washed with hoţ ethanol (30ml). The ethanol was evaporated to give a
residue which was further purified by column chromatography using 100-200 mesh
silica gel and 30% Bthyl acetate- Petether as eluent to yield 13b (70 mg) as a pale
yellow liquid.
HPLC Conditions :
Column : Symmetry SheildRP-18(4.6xl50)mm
Max: Mobile phase : 0.01 M KH2PO4 (PH = 2.5) : Acetonitrile (45:55)
Flowrate : 1. 0 ml/ min ; Wavelength : 270 nm

Retention Urne : 8.99 min ; Purity : 92.24%
IR(KBr, vmax) : 3781,3377,2962,1728,1602,1267,1170,738 cm-1. 1H NMR i (DMSO-d6,300 MHz); 6 7.5(d, 2H), 6.8(d, 2H), 4.3(m, 1H), 3.9(m, 2H), 3.8(m,lH),2.6(2dd,2H),1.9(m,lH), 1.3-1.5(m,4H), 0.8(m,9H). Mass:m/z. 320(M+1), 302,248,192.
To compound lOfo (0.300 g) was added thionyl chloride (lml) at O deg and the resultant clear solution stirred at RT for 30 min. The excess thionyl chloride was removed under reduced pressure (lOmm-Hg). To the residue was added isobutyl amine (1.46 g) and the solution was stirred at RT for 2 hrs. The solution was then diluted witti eutyl acetate (5ml) and washed wim water (2x5ml). The organic layer was concentrated te» a residue. The residue was purifîed by column chromatography (60-120 mesh silica gel, 30% Ethyl acetate-Petether) to yield 14b (0.180g)asaliquid
Compound 14b (0.150 g) was dissorved in emanai (15 ml) and then Paladiumon carbon (0.030 g) was added. The solution was hydrogenated using balloon pressure for four hours. The reaction mixture was filtered through a pad of celite and the bed was washed with hoţ ethanol (30ml). The ethanol was evaporated to give a residue which was further purifîed by column chromatography using 100-200 mesh silica gel and 30% Emyl acetate- Petemer as eluent to give 15b (0.080 g) as a pale brown semi solid. HPLC Conditions:
Column : Symmetry Sheild RP-18 (4.6x150) mm
Max: Mobile phase: 0.01M KH2PO4 : Acetonitrile (55:45)
Flowrate : LOml/min ; Wavelength : 210 nm

Retention time : 6.27 min ; Purity : 93.87%
IR ( KBr, v max) :3416,3300,2924,1653,1610,1550,1515,1348,1275, 809
cm-1. 1H NMR: (CDC13,300MHz ); 6 8.2(br.s, 1H), 7.5(d, 2H), 6.8(d, 2H),
6.2(m, 1H), 4.4(m, 1H), 3.8(m, 1H), 3.1(m, 2H), 2.6(2dd, 2H), l .8(m, 1H), 1.3-
1.5(m, 4H), 0.8(m, 9H). MMS : m/z : 319( M+l), 301,200.
EXAMPLE3
(Figure Removed)
Referring now to the Butyl Series reaction scheme in FIG. 3A: To the solution of Chlorooxime derivative (Compound 8,16.74 g) in THF (100 ml) was added tnethylamine (14.2 g) and the solution was cooled to 5 -10 deg. To this solution was added slowly methyl-3-octenoate (5.0 g) and the resultant solution was stirred at RT for 24 hrs. The solvent was removed by distillation and the residue was dissolved in ethyl acetate (lOOml) and washed with water (2x50ml)
followed by brine. The organic layer was dried over anhydrous sodium sulfate, fiheted, and concentrated to a residoe. The TLC shows that two regioisomers were fonned (Compoonds 16a and 16b, 25 g) as a yellow solid. The crude material was taken on to the ester hydrorysis step.
The crude reaction mass (16a and 16b, 25 g) was taken in methanol (200ml) and a 25% sodium hydroxide solution was added. The resuhant solution was refluxed for 2 hrs. The solvent was removed under reduced pressure (lOmm-Hg) and the residue wasdilutedwithwater(100ml)andadjustedtoapH of 2 with bydrochloric acid (2M). The solution was extracted wfth etiryl acetate (2 x 200 ml). The organic layer was again washed whh brine (lOOml) and the resuhant organic layer was dried over anhydroas sodium sulikte, fittered, and concentrated. The mixture was purified by column chromatography (l00-200 mesh silica gel: 40% Eflryl acetate - Petether) to grve 17» (0.700 g) as » white sobd.
The benzylated acid derivative (Compound 17a, 0300 g) was dissolved in
rtharol (60 nd) and palladium cm carbon solution was
hydrogenated under balloon pressure forfour hours. The reaction mixture was fUtered through a pad of cetite and the bed was washed with hoţ ethanol (30 ml). The ethanol was removed under reduced pressure to give a residue which was further purified by column chromatography using 60-120 mesh silica gel and 10% Methanol and Chloroform as eluent to give 18a (0.060 g) as an off-white solid. M.P:174-176C° FIPLC Conditions:
Column : Symmetryshield(4.6xl50)mm
Mobile phase : 0.01M KH2PO4 (2.5): ACN (60:40)

Flowrate : 1.0 ml/min; Wavelength : 225 nm
Retention time : 5.53 min; Purity : 94.35%
IR(KBrvmax) : 3406,2930,1701,1606,1516,1434,1351,1283,1221,1170, 936,828,833,675 cm-1. 1H NMR: (DMSO-d6,300 MHz); 8 10(s.br, 1H), 7.5(d, 2H), 6.9(d, 2H), 4.8(m, 1H), 3.4(m, 1H), 2.5(m, 2H), 1.2-1.4(m, 6H), 0.8(t, 3H). M«M : m/z. 278(M+1), 234,192,65.
To the benzylated acid derivative (Compound 17a, 0.300 g) was added
thionyl chloride (Iml) at O deg and the resuhant clear sohltion stirred at RT for 30
min. The excess of thionyl chloride was removed under reduced pressure (lOmm-
Hg) and to the residue isoburyl akohol (l .6 g) was added and the sohltion stirred at
RT for 2 hrs. The sohltion wtsdilutcrfwithe* washedwith
water (2x5ml) followedby brine. The organic tayer was concentrated to a residue which was further purified by cohmn chromatography (60-120 mesh silica gel, Petemer and Ethyl acetate (10%)) to ghre 19a (0.150 g) as a fiqirid.
(Table Removed)

Reaction Time: 4 hrs Reaction Temperarure : 25 to 30 deg
The benzylated acid derivative (Compound 19a, 0.150 g) was dissolved in ethanol (l 5 ml) and 10% palladium on carbon was added. The solution was
hydrogenated using balloon pressure for 4 hrs. The reaction mixture was filtered
through a pad of celite and the bed was washed with hoţ ethanol (30 ml). The
ethanol was evapoiated to gjve a residue which was further purified by column
chromatography using 100-200 mesh silica gel and 20% Ethyl acetate - Pet.ether
as eluent to give 20a (0.060 g).
HPLC Co»ditions :
Column : Symmetry shield (4.6x150) mm
Mobile phase : 0.01M KH2PO4 (PH = 2.5): Acetonitrile (40:60)
Flowrate : LOml/min ;Wavelength : 270 nm
Rctention time : 7.51 mm ; Purity : 97.19%
W(KBr, vniax) : 3378,2960,1729,1606,1517,1464,1352,1276,1173,993,
888,839 cm-1. IH NMR: (CDC13,300 MHz); 5 7.5(d, 2H), 6.9(d, 2H), 53(br.s,
IH), 4.8(m, IH), 3.8(d, 2HX 3.4(m,lH), 2.6(2dd, 2H), 1.9(m, IH), 1.3-1.6(m, 6H),
0.9(d, 6H),0.8(t, 3H). MM»: mfz. 334(M+1X 192.
To the benzylated acid derivative (Compound 17a, 0.300 g) was added thionyl chloride (l ml) at O deg and the resultant clear solution stiired at RT for 30 min. The excess of thionyl chloride was removed under reduced pressure (lOmm-Hg) and to the residue was added isobutyl amine (1.46 g) and the resultant solution stirred at RT for 2 hrs. The solution was diluted with ethyl acetate (5 ml) and washed with water (2x5ml) followed by brine. The organic layer was concentrated to a residue which was further purified by column chromatography (60-120 mesh silica gel, 20% Ethyl acetate - Petether) to give 21 a (0.180 g) as a liquid.
The benzylated amide derivative (Compound 21 a, 0.150 g) was dissolved in ethanol (15 ml) and 10% palladium on carbon was added. The solution was
hydrogenated using balloon pressure for four hours. The reaction mixture was
filtered tbrough a pad of cetite and the bed was washed with hoţ ethanol (30 ml).
The elfaanol was evaporated to give a residue which was further purified by column
chromatography using 100-200 mesh silica gel and 20% Ethyl acetate - Petether
as eluent to give 22a (0.060 g) as an oily solid.
HPLC CoaditioBs:
Cohmm : Symmetry C-18(4.6x250)mm
Max: Mobile phase : Water: Acetonhrile (40:60)
Flowrate : O.Smlmin ;Wavelength : 210 nm
Retentiontime : 5.65 min ; Purity : 94.73%
IR (KBr vnuoc): 3298,2959,2930,1646,1607,1517,1461,1353,1278,1172,
888,838cm-l. 1HNMR (CDCB, 300MHz): 6 7.5(d, 2H), 6.8(d, 2H), 6.l(br.s>
1H), 4.8(m, 1H), 3.4(m, 1H), 3.2(im2H), 2.6(2dd, 2H), 1.8(m, 1H), 1.3(m, 4H),
0.8(m, 9H). MM*: m/z. 333(M+1), 315,308,287,286
Referring now to the Butyl Series reaction scheme in FIG. 3B: The solution of ChIorooximederivative(Compound 8,16.74 g) inTHF (100 ml) was added triethylamine (14.2 g) and the solution was cooled to 5 - 10 deg. To this solution was added slowly methyl-3-octenoate (5.0 g) and the resultant solution was stirred at RT for 24 hrs. The solvent was removed by distillation and me residue was dissolved in ethyl acetate (lOOml) and washed with water (2x50ml) followed by brine. The organic layer was dried over anhydrous sodium sulfate, fîltered, and concentrated to a residue. The TLC shows that two regioisomers were formed (Compounds 16a and 16b). The crude yellow solid (25 g) was taken into the hydrolysis step without further purification.
The mixture of 16a and 16b (25.0 g) was taken up in methanol (200ml) and a 25% sodium hydroxide sohrtion (13.6 ml) was added. The resultant solution was refiuxed for 2 hrs. The solvent was removed under reduced pressure (lOmm-Hg) andthe residue wasdiluted withwater (lOOml)andadjustedto apHof 2 with hydrochioric acid (2M). The solution was extracted with ethyl acetate (2 x 200ml). The organic layer was again washed whh brine (lOOml) and the resultant organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture of the two isomers (Compounds 17a and 17b) was further purified by cohman chromatography (100-200 mesh silica gel, : 40% Ethyl acetate - Pet.ether) to give a white solid (1.4 g) of compound 17b.
The benzyhfed acid derivative (Compound 17b„ 0.300 g) was dissorved in etharx)! (60 ml) and 10% palladium on carbon was added. The solution was hydrogenatedimderbaUocti pressure fbrfour hours. The reaction mixture was filtered through abed of cetite and the bed was washed whh hoţ ethanol (30ml). The ethanol was removed under reduced pressure and the residue was further purified by column chromatography using 60-120 mesh silica gel and 10% Methanol and Chloroform as eluent The compound 18b (0.080 g) was isolated as off-white crystals. mp: 163-168°C. HPLC Condiţiona:
Column : SymmetryshieldC-18 (4.6x250) mm
Max: Mobile phase : 0.01M KH2PO4 (PH = 2.5) : Acetonitrile (50:50)
Flowrate : 0.7ml/min ; Wavelength : 270 nm
Retention time : 6.87 min ; Purity : 95.72%

IR (KBr vmax) : 3209,2958,1711,1612, 1597,1519,1434,1352,1274,909, 838 cm-1. 1H NMR: (DMSO -d6,300 MHz); 5 10.0(s.br, 1H), 7.5(d, 2H), 6.9(d, 2H), 4.5(m, 1H), 3.7(m, 1H), 2.5(m, 2H), 1.5(m, 2H), 1.2(m, 4H), 0.8(m, 3H). Mass : M/z. 278(M+1).
To the benzylated acid derivative (Compound 17b, 0.300 g) was added thionyl chloride (Iml) at O deg and the resuttant clear solvrtion was stirred at RT for 30 min. The excess thionyl chloride was removed under reduced pressure (lOmm-Hg). To the residue was added isobutyl alcohol (l .6 g) and the solution stirred at RT for 2 hrs. The solution was dihited with ethyl acetate (5ml) and washed with water (2x5ml) foOowed by brine. The organic teyer was concentratedto a residue. wiuch was further purified by column chromatography (60-120 mesh silica gel, 20%Ethyl»cetiite-Petcther)togivcl9b(0.150g)asaliquki.
The benzySated acid derivative (Compound 19b, 0.150 g) was dissotved in ethanol (15 ml) Inen 10% pdladiumon carbon (0.030 g) was added. The solution was hydrogenated using bAUoon pressure for 4 hrs. The reaction mixturc was fihered over over a bed of celite and the bed was washed with hoţ ethanol (30ml). The ethanol was evaporated to give a residue which was further purified by column chromatography using 100-200 mesh silica gel and 20% Ethyl acetate - Petether as eluent The desired ester 20b (0.060 g) was isolated as a solid. M.P: 114 -116 deg.
HPLC Conditions :
Column : Symmetry sbieldRP-18 (4.6x150) mm
Max: Mobile phase : 0.01M KH2P04 (PH = 2.5) : Acetonitrile (40:60)
Flowrate : 1.0 ml/min ; Wavelength : 270 nm ;
Retentiontime : 8.74 min ; Purity . 97.76%
BR(KBr vmax) : 3159,2958,2870,1734,1614,1596,1519,1445,1354,1273, 1236,1173, 898,838 em-1. IH NMR: (CDCD, 300 MHz); 5 7.5(d, 2H), 6.9(d, 2H), 5.8(br.m, IH), 4.4(m, IH), 3,9(m, 2H), 3.7(m, IH), 2.6(2dd, 2H), 1.9(m, l H), 1.3-1.6(m, 6H), 0.8(m, 9H). Mass: M/z. 334(M+1).
To the benzylated acid derivative (Compound 17b, 0.300 g) was added thionyl chloride (îmi) at O deg and the resuhant clear solution stirred at RT for 30 min. The excess of thionyl chloride was removed under reduced pressure (lOmm-Hg). To the residue was added isobutyl amine (1.46 g) and the resultant solution stirred at RT for 2 hrs. The sohrtion was diluted with ethyl acetate (5ml) and washed wîth water (2x5ml) followed by brine. The organk layer was concentrated to a residue. The Compound 21b was further purifiedby column chromatography (60-120 mesh silica gel, 20% Ethyl acetate - Petether) to grve pure 21b (0.180 g) as a liquid The compound was taken on without further characterization.
The benzyiated amide derivative (Compound 21b, 0.150 g) was dissolved in ethanol (15 ml) then 10% palladium on carbon was added. The solution was hydrogenated using balloon pressure for four hours. The reaction mixture was filtered through a pad of celite and the bed was washed with hoţ ethanol (30ml). The ethanol was evaporated to give a residue. The compound 22b was further purified by column chromatography using 100-200 mesh silica gel and 20% Ethyl acetate - Petether as eluentto yield pure 22b (0.060 g) as a solid. M.P : 157 - 161 deg.
HPLC Conditions:
Column : SymmetryC-18 (4.6 x 150) mm
Max: Mobile phase 0.01MKH2PO4(PH = 2.5) : Acetonitrile (40:60)
Flowrate : l.Omlmin ; Wavelength : 270 nm
Retention time : 10.07 min ; Purity : 91.43%
IR (KBrvmax): 3286,2961,1653,1610,1558,1516,1461,1348,1277,1229,
886, 840 cm-1. IH NMR: (CDC13,300 MHz); 8 7.5(d, 2H), 6.8(d, 2H), 6.1(br.s,
IH), 4.8(m, IH), 3.3(m,lH), 2.6(2dd, 2H), 1.8(m, IH), 1.3(m, 4H), 0.8(m, 9H).
Mass : M/z. 333(M+1), 315,214.
EXAMPLE4
Strnctvre of Target Mofecnles 4

(Figure Removed)
Referring now to the Furyl Series reaction scheme in FIG. 4:

To the solution of Chlorooxime (Compound 8, 15.7 g) in THF (100 ml) was added triethylamme 14.2 g) and the solution was cooled to 5 - 10 deg. To the above sohition was added slowh/ 4-Furan-2-yl-but-3-enoic acid methyl ester (5.0 g) and the resuhant solution was stirred at rt for 24 h. The solvent was removed by distillation and the residue was dissotved in ethyl acetate (lOOml) and washed with water (2x50ml). The organic layer was dried over anhydrous sodium sulfate, fihered, and concentrated to a yeUowish Hquid (4.0 g). The TLC shows that only single isomer îs formed. The crude reaction mass was taken on to the hydrolysis step.
The crude reaction mass (Compound 30, 1.S g) was taken in methanol (20m!) and sodium bydnxtide solution (25%X°-2 g) was added. The resuhant sohition was stirred «t 25 to 30 deg fig 16 hrs. The solvent was removed by
2 with hydrochloric acid (2M). The carboxyhc acid derivative was extracted with ethyl acetate (2 x 200ml) and the organic layer was further washed with brine (lOOml).The combined organic layers were dried over anhydrous sodium sulfate, dried, and concentrated. The compound 31 was further purified by column chromatography 100-200 mesh silica gel, 30% Ethyl acetate - Pet.ether as a solid (800 mg).
The benzylated acid derivative (Compound 31, 0.150 g) was dissolved in ethanol (15 ml) and Palladium on carbon (0.030 g) was added. The solution was hydrogenated using balloon pressure for four hours. The reaction mixture was filtered through a bed of celite and the bed was washed with hoţ ethanol (30ml). The ethanol was evaporated to give a residue. The compound 32 was further
purified by column chromatography using 100-200 mesh silica gel and 40% Ethyl
acetate - Peteiher as eluent to give 32 (0.060 g) as an off-white solid. M.P : 192 -
194°C.
HPLC Conditions :
Column : SymmetryshieldRP-18 (4.6x250)mm
Max: Mobile phase : 0.01M KH2PO4 (PH = 2.5) : Acetonitrile (60:40)
Flowrate : 0.8mL/min.; Wavelength : 210nm
Retention time : 6.15min. ; Purity : 97.82%
IR (KBr. v max) : 3149,2923,1711,1612,1592,1437,1352,1283,911, 837,745
cm-1. 1H NMR: (DMSO 7.6(s,lH), 7.5{cUHX 6.8CdHX 6UH), 5.4(d,lH), 4.2(m, 1H), 2.6(m, 2H).
Muc: m/z, 288(M+1X 270,242,192,164,97.
To the benzylated acid derivative (Compound 31,0.300 g) was added tbionylcntodde(lnri)atOdegandtheresultantclearsota 30 min. The excess thionyl chloride was removed under reduced pressure (lOmm-Hg). To the residue was added isobutyl alcohol (1.6 g) and the solution was stirred at RT for 2 hrs. The solution was diluted with ethyl acetate (5ml) and washed with water (2x5ml). The organic layer was concentrated to a residue purified by column chromatography (60-120 mesh silica gel, 20% Ethyl acetate - Petether). The product 33 was isolated as a liquid (0.180 g).
The benzylated acid derivative (Compound 33,0.200 g) was dissolved in ethanol(15 ml) added Palladium carbon. The reaction mixture was hydrogenated using balloon pressure for fbur hours. The reaction mixture was filtered through a pad of celite and the bed was washed with hoţ ethanol (30 ml). The ethanol was
evaporated to give a residue which was further purified by column chromatography
usii 100-200 mesh silica gel and 40% Emyl acetate - Petether as eluent to give
34 as an off-white solid (0.80 g). M.P: 123 - 125°C.
HPLC Conditions:
Column : SymmetryC-18 94.6x250) mm
Max: Mobile phase : 0.01M KH2PO4 : Acetonitrile (50:50)
Flowrate : l.OmLAnin.; Wavelength : 215nm
Retention tirne : 13.58 min. ; Purity : 95.97%
IR (KBr. v max) : 3781,3221,1735,1598,1517,1440,1348,1276,1228,1175,
736 cm-L 1H NMR: (CDCB, 300 MHZ), 6 7.5(d, 2H), 73(s> 1H), 6.9(d, 2H),
6.5 1H), ©.«(d, 6H). MM : m/z 344(M+1X 326,248,192,151.
Totbcb The benzylated acid derivative (35) was dissolved in ethanol (15 ml) and Palladium on carbon (10% w/w, 0.030mg) was added. The solution was hydrogenated using balloon pressure for 4 hours. The reaction mixture was filtered
through a bed of celite and the bed was washed with bot ethanol (30ml). The
ethanol was evaporated to gjve a residue which was further purified by column
chromatography using 100-200 mesh silica gel and 40% Ethyl acetate - Petether
as eluent to give 36 (0.60 g) as an off-white solid. M.P : 167 - 169°C .
HPLC Conditions:
Column : Symmetry C-18 (4.6 x!50) mm
Max: Mobile phase : 0.01 M KH2PO4 (PH = 2.5) : Acetonitrile (60 :40)
Flowrate : l.OmL/min.; Wavelength : 210nm
Retentionnme : 8.05 min; Purity : 98.17%
ER(KBr. v max) : 3286,2961,1653,1608,1516,1350,1278,1231,1167, 841,
748. 1H NMR: (CDC13,300 MHz), 8 73(d, 2H), 73(s> 1H), 6.9(d, 2H), 6.3(d,
2HX 5.4(br.s, 1H), 53(d, 1H), 430», 1H), 3.1(m, 2H), 2.6(m, 2H), 1.8(m, 1H),
0.8(m, 6H). MM» : m/z. 343(M+1X 325,275,224.
The activity of mc compounds of the invention for the various disorders described above can be detennined according to oue or mare of the following assays. Materials and Methods
Synthesis. In the examples of the syntheses that follow, all reagents and solvents used were purchased at the highest commercial quality. AU solvents used were HPLC grade lBx>m Fisher. 1H (270 MHz) and "CNMR (67.5 MHz) NMR spectra were recorded on a JEOL Eclipse 270 spectrometer. Coupling constants were reported in Hertz (Hz), and chemical shifts were reported in parts per million (ppm) relative to tetramethylsilane (TMS, 0.0 ppm) with CDClj, DMSO or CD3OD as solvent. Thin layer (TLC) and flash column chromatography were performed
using Alumina B, F-254 TLC plates from Selecto Scientific and Fisher Scientific Basic alumina Brockman activity I, respectivery. The reactions were monitored by TLC and 'HNMR and were stopped when the yield of the crude according to 'HNMR was 90-95%.
Reagents. Unless otherwise indicated, aii chemicals were purchased from Aldrich or Sigma Chemical Companies, and were of the highest grade commercially available. Dopachrome methyl ester was prepared similarly to previousry published procedures (Bendrat et al.. Biochemistry, 36,15356-15362 (1997); Swope.etal.. EMBO J„ 17, 3534-3541 (1998)).
Assays were initiated at a time when mc absorbance ai 475 nm reached a maximal vame, sjgmrytngthatthe hmitmgieagent, NaIO4,was consumed. Recombinant human and moaşe MIF was expiessed in E. coti and purified as previousry reported (Bemhagan,, ţţ * Bkxdkemistry, 33, 14144-14155 (1994). MIF Tautornerasc
The compounds of Formula I or II are identified as KfîF inhibitors because they inhibit MIF enzymatic activity in vitro. MIF catalyzes a tautomerization (i.e., keto-enol isomerization) reaction (Rosengreru et al.. Molecular Medicine, 2, 143-1 49 (l 996). MIF catalyzes the tautomerization of a dopachrome-related MIF substrate to a colorless product Unless specifically indicated to the contrary, references made herein to an inhibitory concentration (e. g., ICX or other activity index) refer to the inhibitory activity of a test compound in an MIF tautomerase assay (as further described in detail below, and in Bendrat et al.. Biochemistry, 36, 15356-15362 (1997). The most active substrate identified is a non-physiological D-isomer of dopachrome. This reaction predicts therapeutic MIF inhibitors (see
U.S. Patent No. 6,420,188 and U.S. Patent No. 6,599,938, the disclosures of which are încorporated herein by referencc in tbeir cntirety), InhibitionofMIF tautomerase activity is predictive of inhibition of MIF biologica! activity.
A method for performing an assay for MIF dopachrome tautomerase activity begins with the preperation and oxidation of a DOP A-related substrate precursor, such as L-3,4-dihydroxyphenylalaniiie methyl ester. This oxidation with sodium periodate generates the corresponding dopachrome derivative (e. g., L-3,5-dihYdro-6-hydroxy-5-oxo-2H-indole-2-carboxylic acid methyl ester ("dopachrome methyl ester") that is orange-colored and comprises a convenient substrate for use in a photometric assay fot the enzymatic activity of MIF as a tautomerase. MIF (typicaîly a purified prepantion of recombînaiitMIFatafinalconcentrationof 50-
substrate to a colorless 5>6Kfinvdroxytndoie~2-caboxyh*c acid methyl ester product The enzymatic actrvhy of MIF is measured as the rate of dc-colorization of the colored sohition of the dopachrome-felated substrate in a suitaMe buffer, typicaîly at a time 20 seconds ader additkm of the final assay component and mixing. The absorbance is measured at about 475 nm (or 550 nm for substrate concentrations in excess 0.5 nM). A test compound may be included in the assay solution such that the effect of the test compound on MIF tautomerase activity (i. e., as an inhibitor) may be measured by noting the change in kinetics of substrate tautomerization compared to control assays performed in the absence of the test inhibitor compound. In particular, the MIF tautomerase assay may be conducted essentially as follows:
L-3,4-dihydroxyphenylalaniDe methyl ester (e.g., Sigma D-1507) is a dopachrome aubstrate precursor, and is prepared as a 4 mM solution in dd HjO. Sodium periodate is prepared as an 8 mM solution in dd H2O. Assay Buffer (50 mM potassium phosphate/1 mM EDTA, pH 6.0) is prepared. Purified recombinant MBF is prepared in 150 mM NaCl/20 mM Tris buffer (pH 7.4) as a stock solution convenient to suppry MEP at a final concentration of about 700 ng/ml. Immediately prior to iniuatmg the assay, 3.6 ml dopachrome substrate precursor solution» 2.4 ml periodate solution and 4.0 ml Assay Buffer are combined into a homogeneous mixture (this preparation of dopachrome substrate is suitable for assay use after l mmaixlforaboutBOmmthereaftar). Test compound (typicalry prepared as a «>ncentratedstockmDMSO)andMIFaretbracombinedwithOJmlAss Buffer plus 0.3 ml dopachrome substrate solution to provide the desired final concentration of the test compound in a homogerieous mixture, and the optical density (absorbance) of Uns assay mixture is monhored at 475 nm. Typicalry, ODm is recorded every 5 sec for 0-60 sec, and the OD, for a grventime point is compared to parallel assays where MIF is not added or the test compound is omitted. Inhibition of MIF tautomerase activity by the test compounds is detennined by inhibition of the de-colorization of the assay mixture, often at the 20 sec time point. ICW values for compounds with MIF tautomerase inhibitory activity, corresponding to the concentration of inhibitor that would inhibit MIF tautomerase activity by 50%, are detennined by interpolation of the results from MIF tautomerase assays at severa! different inhibitor concentrations. These IC50 values provide a reasonable correlation between MIF enzymatic inhibitory activity of the test compounds, and inhibition of the biologica! activity of MIF.
The MIF tautomerase assay shows that certam compounds inhibit MIF enzymatic actrvity. The data pcovides a reasonable correlation between the MIF tautomerase enzymatic assay and MIF antagonism in a biologica! assay. Collectivery, these data show mat inhibition by a compound in the MIF tautomerase assay is predictive of its potenţial therapeutic use in inhibiting MIF biologica! activity. mbibition of MIF is also reasonably correlated to the modulation of other cytokines affected by MIF and the ERK/MAPK pathway. Treatment of MIF whfa Inhibitors.
MIF samples were treated whh various concentrations of the inhibitors and treated MIF samples were then anaryzed fior enzyme activity using the dopachrome tautomerase assay. Dopacnromc Tautomerase Assavs.
To a room tempenture somtion of recombinant mouse or human MEF samples was added dopacbrome metbyl ester. The sample was immediateiy monitored for loss in absorbance at 475 mn compared to untreated MIF solutions and to dopachrome methyl ester without the addition of MIF. Enzyme Inhibitipn Studies.
This assay illustrates the inhibition of the enzymatic activity of human MIF by the compounds of the invention. The enzymatic tautomerization activity of recombinant human MIF was performed using L-dopachrome methyl ester as a chromogenic substrate (Bendrat et al.. Biochemistry, 36,15356-15362 (1997)). The tautomerization reaction catalyzed by MIF, as described in detail above, leads to the formation of a dihydroxyindole product which is colorless.
Severa! compounds were prepared and tested for acti vity in the MIF dopachrome tautomerase assay. Compounds 68 and 69 (TABLE I) inhibited MIF tautomerase activity in a dose-dependent manner with an IC of about 10 uM.
Thus, according to the present invention, the compounds related in structure to compound 68 and 69 comprise a new and general class of low molecular weight, specific inhibitors of MIF enzymatic activity. Biologica! Assay of MIF Activity.
This assay shows mat the compounds not only specifically inhibit MIF enzymatic activity, but also inhibit MO7 immunoregulatory actrvities, specifically, MBP glucocorticoid regulating activity. The abih'ty of compounds according to the inventkm to neutraHze the effect of MIF to inftuence the anti-mflammatory erfect oo Tha production by mnian monocytes is tested. The property of the compound is dote dependent Toaddress the specificity of tiusinhiDitory effect on MBF, other analogs arc tested mat arc not such potent inhibitors of MIF tautomerase activity.
These results are consistent with a hypothesis mat the pro-inflammatory effects of MIF can be neutralized by the binding of a small molecule at the tautomerase active site, although this effect is not believed to depend on the neutralization of tautomerase activity per se.
The compounds are additionally assessed for inhibition of MIF biologica! activities in any of a number of assays for MIF biologica! activity including, for example, inhibition of MIF binding to target cells, inhibition of MIF release or synthesis, inhibition of MIF immunoreactivity with MIF-specific antibodies, alterations of MIF conformation or structura! integrity as assessed by circular
dichroism spectroscopy, liquid NMR-spectroscopy, X-ray crystallography, thennal stabtlity measurement, inhibitian of the pro-proliferative effects of NflF on quiescent NIH/3T3 cells and inhibition of the associated prolonged ERK activation therein, inhibition of NflF-induced arachadonic acid release from NIH/3T3 cells, inhibition of MIF-induced fructose 2,6 bisphosphate formation in L6 myocytes, inhibition of MEF toxicity in the MDF, TNF, or LPS-challenged test animals, inhibition of ti» glucocorticoid coanter-regulatory activity of KHF in vitro or in vrvo, inhibition of the MIF-induced funcţional inactivation of the p53 tumor suppressor protein (Hudson. ct al.. L Exp. Med, 190,1375-1382 (1999), inhibition of MIF-induced release ofprostagtondm E2, and inhibition of morbidity or mortality in aay of a mmber of animal models of human diseases that are charactenzed by the letease, prodnctim and/or appearance of MIF.
From the fixegomg deauîption, it cm be seen tiiat the preseut invention comprises a new and unique compounds, composhions, processes of making and inethodsofuserelaAedtoinhflMticfiofMnbyabovecoinpounds. Itwillbe recognized by those skilled na tibe art that changes could be made to the , above-described embodiments of the invention without departing from the broad inventive concepts thereof. It is understood, therefore, that this invention is not limited to the particular embodiments disclosed, but is intended to cover all modifications which are within tite spirit and scope of the invention and that this invention is not limited to the particular embodiments disclosed, but it is intended to cover any modifications which are within the spirit and scope of the present invention.


WE CLAIM;
1. A compound having Formula I:
(Formula Removed)
wherein B is oxygen; and
each R is independently defined as follows:
(Formula Removed)
wherein at least one R in Formula 1 has the following Formula III:
(Formula Removed)
wherein each R1 is independently hydrogen, an alkyl group, a cycloalkyl group, a halo group, a perfluoroalkyl group, a perfluoroalkoxy group, an alkenyl group, an alkynyl group, a hydroxy group, an oxo group, a mercapto group, an alkylthio group, an alkoxy group, an aryl
group, a heteraryl group, an aryloxy group, a heteroaryloxy group, an aralkyl group, a heteroaralkyl group, an aralkoxy group, a heteroaralkoxy group, an HO—(C =O)— group, an amino group, an alkylamino group, a dialkylamino group, a carbamoyl group, an alkylcarbonyl group, an alkoxycarbonyl group, an alkylaminocarbonyl group, a dialkylamino carbonyl group, an arylcarbonyl group, an aryloxycarbonyl group, an alkylsulfonyl group, or an arylsulfonyl group, wherein two independently chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which they are attached to form a three to forty membered, cyclic heterocyclic or heteroaryl ring;
each R2 is independently an alkyl group, a cycloalkyl group, a halo group, a perfluoroalkyl group, a perfluoroalkoxy group, an alkenyl group, an alkynyl group, a hydroxy group, an oxo group, a mercapto group, an alkylthio group, an alkoxy group, an aryl group, a heteroaryl group, a heterocyclic group, an aryloxy group, a heteroaryloxy group, an aralkyl group, a heteroaralkyl group, an aralkoxy group, a heteroaralkoxy group, an HO-(C =O)— group, an amino group, an alkylamino group, a dialkylamino group, a carbamoyl group, an alkylcarbonyl group, an alkoxycarbonyl group, an alkylaminocarbonyl group, a dialkylamino carbonyl group, an arylcarbonyl group, an aryloxycarbonyl group, an alkylsulfonyl group, or an arylsulfonyl group;
each m is independently zero or an integer from one to twenty; and each X is carbon, and each Y is defined independently as follows:
(Formula Removed)
wherein each Z is independently hydrogen, an alkyl group, a cycloalkyl group, a halo group, a perfluoroalkyl group, a perfluoroalkoxy group, an alkenyl group, an alkynyl group, a hydroxy group, an oxo group, a mercapto group, an alkylthio group, an alkoxy group, an aryl group, a heteroaryl group, an aryloxy group, a heteroaryloxy group, an aralkyl group, a heteroaralkyl group, an aralkoxy group, a heteroaralkoxy group, an HO—(C =O)— group, an amino group, an alkylamino group, a dialkylamino group, a carbamoyl group, an alkylcarbonyl group, an alkoxycarbonyl group, an alkylaminocarbonyl group, a dialkylamino carbonyl group, an arylcarbonyl group, an aryloxycarbonyl group, an alkylsulfonyl group, or an arylsulfonyl group;and
each n is independently zero or an integer from one to four;
pharmaceutically acceptable salts thereof and pharmaceutically acceptable prodrugs thereof;
with the proviso that the compound of Formula I does not have the following Formula IA:
(Formula Removed)
wherein in Formula IA:
each Y1 is independently hydrogen or (C1-C6)alkyl;
each Y2 is independently Y1, hydroxyl, halo, —SH, or—N(Y1)2;
Resa is independently Y1, halo, —OY1,—N(Y1)2,—SH, =O, =CH2,or A;
Resb is defined as follows:
(Formula Removed)
Y3 is independently Y1, A, - (CH2)-A, -N(Y1)2, or -NY1 Y5,
A is independently phenyl or an aromatic ring substituted with one or more independent Y2 substituents,
Y5 is independently a saturated or unsaturated, straight or branched (C2-C18)alkyl; and Y4 is independently a Y1, -OY1, -OY5, -N(Y1)2, -NY1 Y5, or A.
2. The compound as claimed in claim 1, which is compound having Formula I, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable prodrug thereof.
3. The compound as claimed in Claim 1, wherein Ar is one of the following:
4. The compound as claimed in claim 1, wherein Ar is:
(Formula Removed)
wherein X and Y are defined above.
5. The compound as claimed in claims 1, wherein R and R1 are each independently selected from the group consisting of hydrogen, (C3-C20)cycloalkyl, (C1-C20)alkoxy, (C1-C20)alkyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic and (C3-C10)cycloalkyl; wherein each of the aforesaid (C1-
C20)alkyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic and (C3-C20)cycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(C1-C6)alkyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic, (C3-C10)cycloalkyl, hydroxy, (C1-C6)alkoxy, perhalo(C1-C6)alkoxy, phenoxy, (C1-C10)heteroaryl-O—, (C1-C10)heterocyclic-O—, (C3-C10)cycloalkyl-O—, (C1-C6)alkyl-S—; wherein two independently chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which they are attached to form a three to forty membered, cyclic heterocyclic or heteroaryl ring.
6. The compound as claimed in claim 1. wherein R and R1 are each independently selected from the group consisting of hydrogen, (C3-C20)cycloalkyl, (C1-C20)alkoxy, (C1-C20)alkyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic and (C3-C10)cycloalkyl; wherein each of the aforesaid (C1-C20)alkyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic and (C3-C2o)cycloalkyl substituents may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(C1-C6)alkyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic, (C3-C10)cycloalkyl, hydroxy, and (C1-C6)alkoxy.
7. The compound as claimed in claim 1, wherein R and R1 are each independently selected from
the group consisting of hydrogen, (C3-C10)cycloalkyl, (C1-C10)alkoxy, (C1-C10)alkyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic and (C3-C10)cycloalkyl.
8. The compound as claimed in claim 1, wherein each R and R1 are defined as independently selected from the group consisting of hydrogen, (C3-C6)cycloalkyl, (C1-C6)alkoxy, and (C1-C6)alkyl.
9. The compound as claimed in claim 1, wherein each R1 is independently selected from the group consisting of hydrogen, (C3-C20)cycloalkyl, (C1-C20)alkoxy, (C1-C20)alkyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic and (C3-C10)cycloalkyl; (C1-C10)heteroaryl-O—, (C1-C10)heterocyclic-O—, (C3-C10)cycloalkyl-O—, (C1-C6)alkyl-S—, (C1-C6)alkyl-SO2—, (C1-C6)alkyl-NH—SO2—, —NO2, amino, (C1-C6)alkyl-amino, [(C1-C6)alkyl]2-amino, (C1-C6)alkyl-SO2—NH—, (C1-C6)alkyl-(C=O)—NH—, (C1-C6)alkyl-(C=O)—[((C1-C6)alkyl)-
N]—, phenyl-(C=O)—NH—, phenyl-(C=O)—[((C1-C6)alkyl)-N]—, —CN, (C1-C6)alkyl-(C=O)—, phenyl-(C=O)—, (C1-C10)heteroaryl-(C=O)—, (C1C10)heterocyclic-(C=O)—, (C3-C10)cycloalkyl-(C=O)—, HO—(C=O)—, (C1—C6)alkyl-O—(C=O)—, H2N(C=O)—(C1 -C6)alkyl-NH—(C=O)—, [(C1-C6)alkyl]2 —N—(C=O)—, phenyl-NH—(C=O)—, phenyl-[((C1-C6)alkyl)-N]—(C=O)—, (C1-C10)heteroaryl-NH—(C=O)—, (C1-C10)heterocyclic-NH—(C=O)—, (C3-C10)cycloalkyl-NH—(C=O)—, (C1-C6)alkyl-(C=O)—O— and phenyl-(C=O)—O—; wherein each of the aforesaid (C1-C20)alkyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic and (C3-C20)cycloalkyl substituents for R1 may optionally be substituted by one to four moieties independently selected from the group consisting of halo, (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(C|-C6)alkyl, phenyl, (C1-C10)heteroaryl, (C1-

Cl0)heterocyclic, (C3-C10)cycloalkyl, hydroxy, (C1-C6)alkoxy, perhalo(C1-C6)alkoxy, phenoxy, (C1C10)heteroaryl-O—, (C1-C10)heterocyclic-O—, (C3-C10)cycloalkyl-O—, (C1-C6)alkyl-S—, (C1-C6)alkyl-SO2—, (C1-C6)alkyI-NH—SO2—, —NO2, amino, (C1-C6)alkyl-amino, [(C1-C6)alkyl]2-amino, (C1-C6)alkyl-SO2—NH—, (C1-C6)alkyl-(C=O)—NH—, (C1-C6)alkyl-(C=O)—[((C1-C6)alkyl)-N]—, phenyl-(C=O)—NH—, phenyl-(C=O)—[((d-QOalkyO-N]—, —CN, (C1-C6)alkyl-(C=O)—, phenyl-(C=O)—, (C1-C10)heteroaryl-(C=O)—, (C1-C10)heterocyclic-(C=O)—, (C3-C10)cycloalkyl-(C=O)—, HO—(C=O)—, (C1-C6)alkyl-O—(C=O)—, H2N(C=O)—(C1-C6)alkyl-NH—(C=O)—, [(C1-C6)alkyl]2 —N—(C=O)—, phenyl-NH—(C=O)—, phenyl-[((C1-C6)alkyl)-N]—(C=O)—, (C,-C,0)heteroaryl-NH—(C=O)—, (C1-C10)heterocyclic-NH—(C=O)—, (C3-C10)cycloalkyl-NH—(C=O)—, (C1-C6)alkyl-(C=O)—O—and phenyl-(C=O)—O—; and wherein two independently chosen R1 alkyl-containing groups may be taken together with any nitrogen atom to which they are attached to form a three to forty membered cyclic, heterocyclic or heteroaryl ring; and
wherein each R2 is independently selected from the group consisting of hydrogen, hydroxyl, halo, —N3, —CN, —SH, (R1)2—N—, (R3)—O—, (R3)—S—, (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C3-C10o)cycloalkyl, phenyl, (C1-C10)heteroaryl, and (C1-C10)hetero-cyclic; wherein each of the aforesaid (C1-C6)alkyl, (C3-C10)cycloalkyl, phenyl, (C1-C10)heteroaryl and (C1-C10)heterocyclic substituents for R2 may optionally be independently substituted by one to four moieties independently selected from the group consisting of halo, (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, perhalo(C1-C6)alkyl, phenyl, (C3-C10)cycloalkyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic, formyl, —CN, (C1-C6)alkyl-(C=O)—, phenyl-(C=O)—, HO-(C=O)—, (C1-C6)alkyl-O—(C=O)—, (C1-C6)alkyl-NH—(C=O)—, [(C1-

C6)alkyl]2—N—(C=O)—, pheny!-NH—(C=O)—,phenyl-[((C1-C6)alkyl)-N]—(C=O)—, —N02j amino, (C1-C6)alkylamino, [(C1-C6)alkyl]2-amino) (C1-C6)alkyl-(C=O)—NH—, (C,-C6)alkyl-(C=O)—[((C1-C6)alkyl)-N]—,phenyl-(C=O)—NH—,phenyl-(C=O)—[((Ci-C6)alkyl)-N]—, H2N—(C=O)—NH—, (C1-C6)alkyl-HN—(C=O)—NH—, [(C1-C6)alkyl-]2N—(C=O)—NH—, (C1-C6)alkyI-HN—(C=O)—[((C1-C6)alkyl)-N]—, [(C1-C6)alkyl-]2N—(C=O)—[((C1-C6)alkyl)-N]—, phenyl-HN—(C=O)—NH—, (phenyl-)2N—(C=O)—NH—, phenyl-HN—(C=O)—[((C1-C6)alkyl)-N]—, (phenyl-)2N—(C=O)—[((C1-C6)alkyl)-N]—, (C1-C6)alkyl-O—(C=O)—NH—, (C1-C6)alkyl-O—(C=O)—[((C1-C6)alkyl)-N]—, phenyl-O—(C=O)— NH—, phenyl-O—(C=O)—[((C1-C6)alkyl)-N]—, (C1-C6)alkyl-SO2NH—, phenyl-SO2NH—, (C1-C6)alkyl-SO2—, phenyl-SO2—, hydroxy, (C1-C6)alkoxy, perhalo(C1-C6)alkoxy, phenoxy, (C1-C6)alkyl-(C=O)—O—, phenyl-(C=O)—O—, H2N—(C=O)—O—, (C1-C6)alkyl-HN—(C=O)—O—, [(C1-C6)alkyl-]2N—(C=O)—O—, phenyl-HN—(C=O)—O—, (phenyl-)2 N—(C=O)—O—; wherein when said R2 phenyl contains two adjacent substituents, such substituents may optionally be taken together with the carbon atoms to which they are attached to form a five to six membered carbocyclic or heterocyclic ring; wherein each of said moieties containing a phenyl alternative may optionally be substituted by one or two radicals independently selected from the group consisting of (Cj-C6)alkyl, halo, (C1-C6)alkoxy, perhalo(C1-C6)alkyl and perhalo(C|-C6)alkoxy.
10. The compound as claimed in claim 1, having the formula:

(Formula Removed)
wherein Rx is a (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic or (C3-C10)cycloalkyl group.
11. The compound as claimed in claim 1, having the formula:
(Formula Removed)
wherein Rx is a (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic or (C3-C10)cycloalkyl group.
12. The compound as claimed in claim 1, having the formula:
(Formula Removed)
wherein Rx is a (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, phenyl, (C1-C10)heteroaryl, (C1-C10)heterocyclic or (C3-C10)cycloalkyl group.
13. The compound as claimed in claim 1, having the formula:
(Formula Removed)
14. The compound as claimed in claim 1, having the formula: E
(Formula Removed)

Documents:

5224-DELNP-2006-Abstract-(20-07-2012).pdf

5224-delnp-2006-abstract.pdf

5224-DELNP-2006-Claims-(20-07-2012).pdf

5224-delnp-2006-claims.pdf

5224-DELNP-2006-Correspondence Others-(20-07-2012)..pdf

5224-delnp-2006-Correspondence Others-(20-07-2012).pdf

5224-delnp-2006-correspondence-others (24-03-2008).pdf

5224-delnp-2006-correspondence-others (31-03-2008).pdf

5224-delnp-2006-correspondence-others.pdf

5224-delnp-2006-description (complete).pdf

5224-DELNP-2006-Drawings-(20-07-2012).pdf

5224-delnp-2006-drawings.pdf

5224-DELNP-2006-Form-1-(20-07-2012).pdf

5224-delnp-2006-form-1.pdf

5224-delnp-2006-form-13 (31-03-2008).pdf

5224-delnp-2006-form-18 (24-03-2008).pdf

5224-DELNP-2006-Form-2-(20-07-2012).pdf

5224-delnp-2006-form-2.pdf

5224-delnp-2006-form-26.pdf

5224-DELNP-2006-Form-3-(20-07-2012).pdf

5224-delnp-2006-form-3.pdf

5224-delnp-2006-form-5.pdf

5224-DELNP-2006-GPA-(20-07-2012).pdf

5224-delnp-2006-gpa.pdf

5224-delnp-2006-pct-304.pdf

5224-delnp-2006-pct-311.pdf

5224-delnp-2006-Petition-137-(20-07-2012).pdf


Patent Number 256046
Indian Patent Application Number 5224/DELNP/2006
PG Journal Number 17/2013
Publication Date 26-Apr-2013
Grant Date 24-Apr-2013
Date of Filing 11-Sep-2006
Name of Patentee CYTOKINE PHARMASCIENCES, INC.
Applicant Address 150 SOUTH WARNER ROAD, SUITE 420, KING OF PRUSSIA, PA 19406, U.S.A.
Inventors:
# Inventor's Name Inventor's Address
1 SIELECKI, THAIS 900 MERRYBELL LANE, KENNETT SQUARE, PA 19348, U.S.A.
2 CRUZ, VIDAL DE LA 101 BRIMFUL DRIVE, PHOENIXVILLE, PA 19460, U.S.A.
PCT International Classification Number A61K 31/42
PCT International Application Number PCT/US2005/010444
PCT International Filing date 2005-03-28
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 60/556,440 2004-03-26 U.S.A.