HK1152712B - Compounds including an anti-inflammatory pharmacore and methods of use - Google Patents

Description

Compounds comprising anti-inflammatory pharmacophores and methods of use

Technical Field

This application claims priority to U.S. provisional application No. 61/046,363, filed on 18.4.2008, which is incorporated by reference in its entirety.

I. Field of the invention

The present invention relates generally to the fields of biology and medicine. More particularly, it relates to compounds and methods for the treatment and prevention of diseases, such as those associated with oxidative stress and inflammation.

Description of the related Art

Many serious and intractable human diseases are associated with a deregulation of inflammatory processes, including diseases such as cancer, atherosclerosis and diabetes, which are not traditionally considered inflammatory diseases. Similarly, autoimmune diseases such as rheumatoid arthritis, lupus, psoriasis and multiple sclerosis involve inappropriate and chronic activation of inflammatory processes in affected tissues, resulting from dysfunction of self-to non-self recognition and response mechanisms in the immune system. In neurodegenerative diseases such as alzheimer's disease and parkinson's disease, nerve damage is associated with activation of microglia and increased levels of pro-inflammatory proteins such as Inducible Nitric Oxide Synthase (iNOS).

One aspect of inflammation is the production of inflammatory prostaglandins, such as prostaglandin E, the precursor of which is produced by COX-2. COX-2 is found at high levels in inflamed tissues. Thus, inhibition of COX-2 is known to reduce many symptoms of inflammation, and many important anti-inflammatory drugs (e.g., ibuprofen and celecoxib) act by inhibiting COX-2 activity. However, recent studies have demonstrated that a class of cyclopentenone prostaglandins (e.g., 15-deoxy prostaglandin J2, also known as PGJ2) plays a role in stimulating the resolution of inflammation. COX-2 is also associated with the production of cyclopentenone prostaglandins. Thus, inhibition of COX-2 may interfere with complete resolution of inflammation, potentially promoting retention of activated immune cells in tissues and leading to chronic "smoldering" inflammation. This effect may lead to an increased incidence of cardiovascular disease in patients who use selective COX-2 inhibitors over a prolonged period of time. Another important class of anti-inflammatory corticosteroids have many adverse side effects and are often not suitable for chronic use. Newer protein-based drugs, such as anti-TNF monoclonal antibodies, have proven effective for the treatment of certain autoimmune diseases, such as rheumatoid arthritis. However, these compounds must be administered by injection, are not effective in all patients and may have serious side effects. In many severe forms of inflammation (e.g. sepsis, acute pancreatitis), current drugs are ineffective. Furthermore, currently available drugs do not have significant antioxidant properties and are not effective in reducing oxidative stress associated with the overproduction of reactive oxygen species and related molecules such as peroxynitrite. Thus, there is an urgent need for improved therapeutic agents with antioxidant and anti-inflammatory properties.

Synthetic oleanolic acid triterpenoid analogs have been shown to be inhibitors of cellular inflammatory processes, such as Inducible Nitric Oxide Synthase (iNOS) and the induction of COX-2 by IFN- γ in mouse macrophages. See Honda et al (2000 a); honda et al (2000b), and Honda et al (2002), all of which are incorporated herein by reference. For example, one of 2-cyano-3, 12-dioxoolean-1, 9(11) -diene-28-oic acid methyl ester (CDDO-Me) is currently used in clinical trials for a variety of inflammation-related disorders, including cancer and diabetic nephropathy. Synthetic another triterpenoid derivative, betulinic acid, has been shown to inhibit cellular inflammatory processes, although these compounds are not widely characterized (Honda et al, Bioorg Med Chem Lett.2006; 16 (24): 6306-9). The pharmacology of these synthetic triterpenoid molecules is complex. It has been demonstrated that compounds derived from oleanolic acid affect the function of multiple protein targets and thereby modulate the activity of several important cellular signaling pathways associated with oxidative stress, cell cycle control and inflammation (e.g., Dinkova-Kostova et al, Proc Natl Acad Sci USA.2005; 102 (12): 4584-9; Ahmad et al, J.biol chem.2006; 281 (47): 35764-9; Ahmad et al, Cancer Res.2008; 68 (8): 2920-6; Liby et al, Nat Rev cancer.2007; 7 (5): 357-69). Betulinic acid derivatives have been shown to have comparable anti-inflammatory properties, but also to show significant differences in pharmacology compared to OA-derived compounds (Liby et al, Mol Cancer Ther 2007; 6 (7)). Furthermore, it is uncertain whether the triterpenoid starting material currently employed has the best properties compared to other possible starting materials. In view of the wide variety of diseases that are known to differ in the spectrum of biological activities of triterpenoid derivatives and in view of the high degree of unmet medical needs that may be treated or prevented with compounds having potent antioxidant and anti-inflammatory effects, as well as the high degree of unmet medical needs that are exhibited in this variety of diseases, it is desirable to synthesize new compounds with diverse structures that may have improved biological activity characteristics for the treatment of one or more indications.

Disclosure of Invention

The present disclosure overcomes the limitations of the prior art by providing novel compounds having antioxidant and anti-inflammatory properties, methods of making the same, and methods of using the same.

In some embodiments, the present disclosure provides a compound comprising:

a) an organic compound having a basic skeleton of 2 to 8 five and/or six membered rings, with the proviso that the basic skeleton is not a basic skeleton of argentatin, betulinic acid, lanostane, oleanolic acid, boswellic acid, glycyrrhetinic acid, ursolic acid, or tricyclic diketene;

b) structural units of the formula:

wherein:

the carbon atoms labeled 1, 2 and 3 are part of a five or six membered ring;

x is cyano or-C (O) R_aWherein R is_aThe method comprises the following steps:

hydrogen, hydroxy, halogen, amino, hydroxyamino, azido or mercapto; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Alkyl sulfonyl amino _(C≤12)Amide group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups; and

R₁and R₂Each is independently:

hydrogen; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Or substituted versions of any of these groups; or

R₁And R₂Are bound together and are alkanediyl_(C≤12)Alkenyldiyl group_(C≤12)Alkanediyl_(C≤12)Or alkenediyl_(C≤12)(ii) a And

c)0 to 8 chemical groups attached to a basic backbone carbon atom other than carbon atoms 1, 2, 3, or 4, wherein each chemical group is independently:

hydroxy, halogen, oxygen, amino, hydroxyamino, nitro, imino, cyano, azido, mercapto or sulfur; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkylene group(s)_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group _(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Alkyl sulfonyl amino_(C≤12)Amide group_(C≤12)Alkyl imino radical_(C≤12)Alkenylimino radical_(C≤12)Alkynylimino radical_(C≤12)An arylimino group_(C≤12)Aralkyl imino radical_(C≤12)Heteroarylimino radical_(C≤12)Heteroarylalkylimino radical_(C≤12)Acylimino group_(C≤12)Alkylthio group_(C≤12)Alkenylthio radicals_(C≤12)Alkynylthio radicals_(C≤12)Arylthio radicals_(C≤12)Aralkylthio group_(C≤12)Heteroarylthio radicals_(C≤12)Heteroarylalkylthio groups_(C≤12)Acylthio groups_(C≤12)Thioacyl, thioacyl_(C≤12)Alkyl sulfonyl group_(C≤12)Alkenylsulfonyl group_(C≤12)Alkynylsulfonyl, and a process for producing the same_(C≤12)Aryl sulfonyl group_(C≤12)Aralkyl sulfonyl group_(C≤12)Heteroaryl sulfonyl group_(C≤12)Heteroarylalkylsulfonyl group_(C≤12)Alkyl ammonium_(C≤12)Alkyl sulfonium_(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups;

or a pharmaceutically acceptable salt, ester, hydrate, solvate, tautomer, acetal, ketal, prodrug, or optical isomer thereof.

In some embodiments, R₁And R₂Each is independently hydrogen, alkyl_(C≤8)Or substituted alkyl_(C≤8). In some embodiments, R₁And R₂Are all hydrogen. In some embodiments, R ₁And R₂Are both methyl groups. In some embodiments, R₁And R₂Are not hydrogen. In some embodiments, the organic compound is not a triterpenoid.

In some aspects, the present disclosure provides a compound comprising:

a) a basic skeleton of a natural product having 2 to 8 rings, with the proviso that the natural product is not argentatin, betulinic acid, lanostane, oleanolic acid, or ursolic acid;

b) structural units of the formula:

wherein:

the carbon atoms labeled 1, 2, and 3 are part of a six-membered ring;

x is cyano, fluoroalkyl_(C≤8)Substituted fluoroalkyl radicals_(C≤8)or-C (O) R_aWherein R is_aThe method comprises the following steps: hydrogen, hydroxy, halogen, amino, hydroxyamino, azido or mercapto; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Alkyl sulfonyl amino_(C≤12)Amide group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups; and

R₁And R₂Each is independently:

hydrogen; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Or substituted versions of any of these groups; or

R₁And R₂Are bound together and are alkanediyl_(C≤12)Alkenyldiyl group_(C≤12)Alkanediyl_(C≤12)Or alkenediyl_(C≤12)(ii) a And

c)0 to 8 chemical groups attached to a basic backbone carbon atom other than carbon atoms 1, 2, 3, or 4, wherein each chemical group is independently:

hydroxy, halogen, oxygen, amino, hydroxyamino, nitro, imino, cyano, azido, mercapto or sulfur; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkylene group(s)_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group _(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Alkyl sulfonyl amino_(C≤12)Amide group_(C≤12)Alkyl imino radical_(C≤12)Alkenylimino radical_(C≤12)Alkynylimino radical_(C≤12)An arylimino group_(C≤12)Aralkyl imino radical_(C≤12)Heteroarylimino radical_(C≤12)Heteroarylalkylimino radical_(C≤12)Acylimino group_(C≤12)Alkylthio group_(C≤12)Alkenylthio radicals_(C≤12)Alkynylthio radicals_(C≤12)Arylthio radicals_(C≤12)Aralkylthio group_(C≤12)Heteroarylthio radicals_(C≤12)Heteroarylalkylthio groups_(C≤12)Acylthio groups_(C≤12)Thioacyl, thioacyl_(C≤12)Alkyl sulfonyl group_(C≤12)Alkenylsulfonyl group_(C≤12)Alkynylsulfonyl, and a process for producing the same_(C≤12)Aryl sulfonyl group_(C≤12)Aralkyl sulfonyl group_(C≤12)Heteroaryl sulfonyl group_(C≤12)Heteroarylalkylsulfonyl group_(C≤12)Alkyl ammonium_(C≤12)Alkyl sulfonium_(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups;

or a pharmaceutically acceptable salt, ester, hydrate, solvate, tautomer, acetal, ketal, prodrug, or optical isomer thereof.

In some embodiments, R₁And R₂Each is independently hydrogen, alkyl_(C≤8)Or substituted alkyl_(C≤8). In some embodiments, R₁And R₂Are all hydrogen. In some embodiments, R₁And R₂Are both methyl groups. In some embodiments, R₁And R₂Are not hydrogen. In some embodiments, the natural product is not boswellic acid or glycyrrhetinic acid. In some embodiments, the natural product is not a triterpenoid. In some embodiments, X is-CN or-C (═ O) NHS (═ O) ₂CH₃. In some embodiments, X is — CN. In some embodimentsIn which X is fluoroalkyl_(C≤8). In some embodiments, X is-CF₃。

In some embodiments, the structural unit is further defined as:

in some embodiments, the basic backbone of the natural product has 2 rings. In some embodiments, the rings are connected to each other by a single chain of atoms. In some embodiments, the single-chain backbone further comprises at least one carbon-carbon double bond.

In some embodiments, the natural product is curcumin. In some embodiments, the compound is further defined by the formula:

wherein:

R₁and R₂Each is independently:

hydrogen; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Or substituted versions of any of these groups; or

R₁And R₂Are bound together and are alkanediyl_(C≤12)Alkenyldiyl group_(C≤12)Alkanediyl_(C≤12)Or alkenediyl_(C≤12)(ii) a And

R₃and R₄Each is independently:

Hydrogen, hydroxy, halogen, amino, hydroxyamino, nitro, cyano, azido or mercapto; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Alkyl sulfonyl amino_(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups;

or a pharmaceutically acceptable salt, ester, hydrate, solvate, tautomer, acetal, ketal, prodrug, or optical isomer thereof. In some variations, R₁And R₂Are both methyl groups.

For example, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof. For example, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In some embodiments, the natural product is resveratrol. In some embodiments, the compound is further defined by the formula:

Wherein:

R₁and R₂Each is independently:

hydrogen; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Or substituted versions of any of these groups; or

R₁And R₂Are bound together and are alkanediyl_(C≤12)Alkenyldiyl group_(C≤12)Alkanediyl_(C≤12)Or alkenediyl_(C≤12)(ii) a And

R₅and R₆Each is independently:

hydrogen, hydroxy, halogen, amino, hydroxyamino, nitro, cyano, azido or mercapto; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy _(C≤12)Or substituted versions of any of these groups;

or a pharmaceutically acceptable salt, ester, hydrate, solvate, tautomer, acetal, ketal, prodrug, or optical isomer thereof. In some embodiments, R₁And R₂Are both methyl groups. For example, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In some embodiments, the basic backbone of the natural product has 3 rings. In some embodiments, the natural product is gallocatechol. In some embodiments, the compound is further defined by the formula:

wherein:

R₁and R₂Each is independently:

hydrogen; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Or substituted versions of any of these groups; or

R₁And R₂Are bound together and are alkanediyl_(C≤12)Alkenyldiyl group_(C≤12)Alkanediyl_(C≤12)Or alkenediyl_(C≤12)；

R₇、R₈And R ₉Each is independently:

hydrogen, hydroxy, halogen, amino, hydroxyamino, nitro, cyano, azido or mercapto; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups; and

R₁₀the method comprises the following steps:

hydrogen, hydroxy, halogen, oxygen, amino, hydroxyamino, nitro, imino, cyano, azido, mercapto or sulfur; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkylene group(s)_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group _(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Alkyl sulfonyl amino_(C≤12)Amide group_(C≤12)Alkyl imino radical_(C≤12)Alkenylimino radical_(C≤12)Alkynylimino radical_(C≤12)An arylimino group_(C≤12)Aralkyl imino radical_(C≤12)Heteroarylimino radical_(C≤12)Heteroarylalkylimino radical_(C≤12)Acylimino group_(C≤12)Alkylthio group_(C≤12)Alkenylthio radicals_(C≤12)Alkynylthio radicals_(C≤12)Arylthio radicals_(C≤12)Aralkylthio group_(C≤12)Heteroarylthio radicals_(C≤12)Heteroarylalkylthio groups_(C≤12)Acylthio groups_(C≤12)Thioacyl, thioacyl_(C≤12)Alkyl sulfonyl group_(C≤12)Alkenylsulfonyl group_(C≤12)Alkynylsulfonyl, and a process for producing the same_(C≤12)Aryl sulfonyl group_(C≤12)Aralkyl sulfonyl group_(C≤12)Heteroaryl sulfonyl group_(C≤12)Heteroarylalkylsulfonyl group_(C≤12)Alkyl ammonium (A), (B), (C) and (C)_C≤12)Alkyl sulfonium_(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups;

or a pharmaceutically acceptable salt, ester, hydrate, solvate, tautomer, acetal, ketal, prodrug, or optical isomer thereof. In some embodiments, R₁And R₂Are both methyl groups. For example, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In some embodiments, the compound is further defined by the formula:

wherein:

R₁and R ₂Each is independently:

hydrogen; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Or substituted versions of any of these groups; or

R₁And R₂Are bound together and are alkanediyl_(C≤12)Alkenyldiyl group_(C≤12)Alkanediyl_(C≤12)Or alkenediyl_(C≤12)；

R₁₁The method comprises the following steps:

hydrogen, hydroxy, halogen, oxygen, amino, hydroxyamino, nitro, imino, cyano, azido, mercapto or sulfur; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkylene group(s)_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Alkyl sulfonyl amino_(C≤12)Amide group_(C≤12)Alkyl imino radical_(C≤12)Alkenylimino radical _(C≤12)Alkynylimino radical_(C≤12)An arylimino group_(C≤12)Aralkyl imino radical_(C≤12)Heteroarylimino radical_(C≤12)Heteroarylalkylimino radical_(C≤12)Acylimino group_(C≤12)Alkylthio group_(C≤12)Alkenylthio radicals_(C≤12)Alkynylthio radicals_(C≤12)Arylthio radicals_(C≤12)Aralkylthio group_(C≤12)Heteroarylthio radicals_(C≤12)Heteroarylalkylthio groups_(C≤12)Acylthio groups_(C≤12)Thioacyl, thioacyl_(C≤12)Alkyl sulfonyl group_(C≤12)Alkenylsulfonyl group_(C≤12)Alkynylsulfonyl, and a process for producing the same_(C≤12)Aryl sulfonyl group_(C≤12)Aralkyl sulfonyl group_(C≤12)Heteroaryl sulfonyl group_(C≤12)Heteroarylalkylsulfonyl group_(C≤12)Alkyl ammonium_(C≤12)Alkyl sulfonium_(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups; and

R₁₂、R₁₃and Rx are each independently:

hydrogen, hydroxy, halogen, amino, hydroxyamino, nitro, cyano, azido or mercapto; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group _(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups;

or a pharmaceutically acceptable salt, ester, hydrate, solvate, tautomer, acetal, ketal, prodrug, or optical isomer thereof. In some embodiments, R₁And R₂Are both methyl groups. For example, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In some embodiments, the basic backbone of the natural product has 4 rings. In another embodiment, the basic backbone of the natural product has 5 rings. In some embodiments, the natural product is celastrol. In a further embodiment, the basic skeleton of the natural product has 6 rings. In some embodiments, the natural product is hecogenin, tigogenin, or sarsasapogenin (sarsaprogenin).

In some embodiments, the compound is further defined by the formula:

wherein:

R₁and R₂Each is independently:

hydrogen; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group _(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Or substituted versions of any of these groups; or

R₁And R₂Are bound together and are alkanediyl_(C≤12)Alkenyldiyl group_(C≤12)Alkanediyl_(C≤12)Or alkenediyl_(C≤12)(ii) a And

R₁₅、R₁₆、R₁₇、R₁₈and R₁₉Each is independently:

hydrogen, hydroxy, halogen, oxygen, amino, hydroxyamino, nitro, imino, cyano, azido, mercapto or sulfur; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkylene group(s)_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Alkyl sulfonyl amino_(C≤12)Amide group_(C≤12)Alkyl imino radical_(C≤12)Alkenylimino radical_(C≤12)Alkynylimino radical_(C≤12)An arylimino group_(C≤12)Aralkyl imino radical_(C≤12)Heteroarylimino radical_(C≤12)Heteroarylalkylimino radical_(C≤12)Acylimino group_(C≤12)Alkylthio group _(C≤12)Alkenylthio radicals_(C≤12)Alkynylthio radicals_(C≤12)Arylthio radicals_(C≤12)Aralkylthio group_(C≤12)Heteroarylthio radicals_(C≤12)Heteroarylalkylthio groups_(C≤12)Acylthio groups_(C≤12)Thioacyl, thioacyl_(C≤12)Alkyl sulfonyl group_(C≤12)Alkenylsulfonyl group_(C≤12)Alkynylsulfonyl, and a process for producing the same_(C≤12)Aryl sulfonyl group_(C≤12)Aralkyl sulfonyl group_(C≤12)Heteroaryl sulfonyl group_(C≤12)Heteroarylalkylsulfonyl group_(C≤12)Alkyl ammonium_(C≤12)Alkyl sulfonium_(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups;

or a pharmaceutically acceptable salt, ester, hydrate, solvate, tautomer, acetal, ketal, prodrug, or optical isomer thereof. In some embodiments, R₁And R₂Are both methyl groups. In some embodiments, R₁₇Is methyl. For example, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In some embodiments, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In one aspect, the present disclosure provides a compound of the formula:

wherein:

R₁and R₂Each is independently:

hydrogen; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group _(C≤12)Acyl group_(C≤12)Alkoxy group_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Amide group_(C≤12)Or substituted versions of any of these groups; or

R₁And R₂Are bound together and are alkanediyl_(C≤12)Alkenyldiyl group_(C≤12)Alkanediyl_(C≤12)Or alkenediyl_(C≤12)(ii) a And

R₂₀、R₂₁and R₂₂Each is independently:

hydrogen, hydroxy, halogen, oxygen, amino, hydroxyamino, nitro, imino, cyano, azido, mercapto or sulfur; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkylene group(s)_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Alkyl sulfonyl amidesBase of_(C≤12)Amide group_(C≤12)Alkyl imino radical_(C≤12)Alkenylimino radical_(C≤12)Alkynylimino radical_(C≤12)An arylimino group_(C≤12)Aralkyl imino radical_(C≤12)Heteroarylimino radical _(C≤12)Heteroarylalkylimino radical_(C≤12)Acylimino group_(C≤12)Alkylthio group_(C≤12)Alkenylthio radicals_(C≤12)Alkynylthio radicals_(C≤12)Arylthio radicals_(C≤12)Aralkylthio group_(C≤12)Heteroarylthio radicals_(C≤12)Heteroarylalkylthio groups_(C≤12)Acylthio groups_(C≤12)Thioacyl, thioacyl_(C≤12)Alkyl sulfonyl group_(C≤12)Alkenylsulfonyl group_(C≤12)Alkynylsulfonyl, and a process for producing the same_(C≤12)Aryl sulfonyl group_(C≤12)Aralkyl sulfonyl group_(C≤12)Heteroaryl sulfonyl group_(C≤12)Heteroarylalkylsulfonyl group_(C≤12)Alkyl ammonium_(C≤12)Alkyl sulfonium_(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups;

or a pharmaceutically acceptable salt, ester, hydrate, solvate, tautomer, acetal, ketal, prodrug, or optical isomer thereof. In some embodiments, R₁And R₂Are all hydrogen. In other embodiments, R₁And R₂Is not hydrogen. In yet another embodiment, R₁And R₂Are both methyl groups.

In some embodiments, the compound is further defined as:

wherein R is₂₀、R₂₁And R₂₂Each is independently:

hydrogen, hydroxy, halogen, oxygen, amino, hydroxyamino, nitro, imino, cyano, azido, mercapto or sulfur; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkylene group(s) _(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Alkyl sulfonyl amino_(C≤12)Amide group_(C≤12)Alkyl imino radical_(C≤12)Alkenylimino radical_(C≤12)Alkynylimino radical_(C≤12)An arylimino group_(C≤12)Aralkyl imino radical_(C≤12)Heteroarylimino radical_(C≤12)Heteroarylalkylimino radical_(C≤12)Acylimino group_(C≤12)Alkylthio group_(C≤12)Alkenylthio radicals_(C≤12)Alkynylthio radicals_(C≤12)Arylthio radicals_(C≤12)Aralkylthio group_(C≤12)Heteroarylthio radicals_(C≤12)Heteroarylalkylthio groups_(C≤12)Acylthio groups_(C≤12)Thioacyl, thioacyl_(C≤12)Alkyl sulfonyl group_(C≤12)Alkenylsulfonyl group_(C≤12)Alkynylsulfonyl, and a process for producing the same_(C≤12)Aryl sulfonyl group_(C≤12)Aralkyl sulfonyl group_(C≤12)Heteroaryl sulfonyl group_(C≤12)Heteroarylalkylsulfonyl group_(C≤12)Alkyl ammonium_(C≤12)Alkyl sulfonium_(C≤12)Alkyl silyl group_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups;

or a pharmaceutically acceptable salt, ester, hydrate, solvate, tautomer, acetal, ketal, prodrug, or optical isomer thereof.

For example, the present disclosure provides:

Or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

For example, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

For example, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In some embodiments, the present disclosure provides a compound selected from the group consisting of:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In some variations, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In some variations, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In another aspect, the present disclosure provides a compound of the formula:

wherein:

R₂₃and R₂₄Each is independently:

or hydrogen, alkyl_(C≤8)Or substituted alkyl_(C≤8)(ii) a Or

R₂₃And R₂₄Are bound together and are alkanediyl_(C≤12)Alkenyldiyl group_(C≤12)Alkanediyl_(C≤12)Or alkenediyl_(C≤12)；

R₂₅，R₂₆And R₂₇Each is independently:

hydrogen, hydroxy, halogen, oxygen, amino, hydroxyamino, nitro, imino, cyano, azido, mercapto or sulfur; or

Alkyl radical_(C≤12)Alkenyl radical_(C≤12)Alkynyl group_(C≤12)Aryl radical_(C≤12)Aralkyl group_(C≤12)Heteroaryl group_(C≤12)Heteroarylalkyl group_(C≤12)Acyl group_(C≤12)Alkylene group(s)_(C≤12)Alkoxy group_(C≤12)Alkenyloxy group_(C≤12)Alkynyloxy, alkynyloxy_(C≤12)Aryloxy group_(C≤12)(iii) aralkyloxy_(C≤12)Heteroaryloxy group_(C≤12)Hetero aralkyloxy group_(C≤12)(iii) acyloxy group_(C≤12)Alkylamino group_(C≤12)Dialkylamino group_(C≤12)Alkoxyamino group_(C≤12)Alkenylamino group_(C≤12)Alkynyl amino group_(C≤12)Arylamino group_(C≤12)Aralkylamino group_(C≤12)Heteroaryl amino_(C≤12)Heteroarylalkylamino, heteroarylalkylamino_(C≤12)Alkyl sulfonyl amino_(C≤12)Amide group_(C≤12)Alkyl imino radical_(C≤12)Alkenylimino radical_(C≤12)Alkynylimino radical_(C≤12)An arylimino group_(C≤12)Aralkyl imino radical_(C≤12)Heteroarylimino radical_(C≤12)Heteroarylalkylimino radical_(C≤12)Acylimino group_(C≤12)Alkylthio group_(C≤12)Alkenylthio radicals_(C≤12)Alkynylthio radicals_(C≤12)Arylthio radicals_(C≤12)Aralkylthio group_(C≤12)Heteroarylthio radicals_(C≤12)Heteroarylalkylthio groups_(C≤12)Acylthio groups_(C≤12)Thioacyl, thioacyl_(C≤12)Alkyl sulfonyl group_(C≤12)Alkenylsulfonyl group_(C≤12)Alkynylsulfonyl, and a process for producing the same_(C≤12)Aryl sulfonyl group_(C≤12)Aralkyl sulfonyl group_(C≤12)Heteroaryl sulfonyl group_(C≤12)Heteroarylalkylsulfonyl group_(C≤12)Alkyl ammonium_(C≤12)Alkyl sulfonium_(C≤12)Alkyl groupSilyl radical_(C≤12)Alkyl siloxy_(C≤12)Or substituted versions of any of these groups;

or a pharmaceutically acceptable salt, ester, hydrate, solvate, tautomer, acetal, ketal, prodrug, or optical isomer thereof.

For example, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

For example, the present disclosure provides:

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer, or optical isomer thereof.

In some embodiments, the present disclosure provides a compound selected from the group consisting of:

(S) -3- ((1E, 6E) -7- (4-hydroxy-3-methoxyphenyl) -4, 4-dimethyl-3, 5-dioxahept-1, 6-dienyl) -3, 5, 5-trimethyl-6-oxocyclohex-1-enenitrile,

(S) -3- ((1E, 6E) -7- (4-hydroxy-3-methoxyphenyl) -5, 5-dimethoxy-4, 4-dimethyl-3-oxahept-1, 6-dienyl) -3, 5, 5-trimethyl-6-oxocyclohex-1-enenitrile,

(S) -3- ((E) -4- (2- (4-hydroxy-3-methoxystyryl) -1, 3-dioxolan-2-yl) -4-methyl-3-oxopent-1-enyl) -3, 5, 5-trimethyl-6-oxocyclohex-1-enenitrile,

(S) -3- ((E) -4- (2- (4- (tert-butyldimethylsilyloxy) -3-methoxystyryl) -1, 3-dioxolan-2-yl) -4-methyl-3-oxopent-1-enyl) -3, 5, 5-trimethyl-6-oxocyclohex-1-enenitrile,

(S, E) -3- (4-hydroxystyryl) -3, 5, 5-trimethyl-6-oxocyclohex-1-enenitrile,

(2S, 3S) -6-cyano-4 a, 8, 8-trimethyl-7-oxo-2- (3, 4, 5-trihydroxyphenyl) -3, 4, 4a, 7, 8, 8 a-hexahydro-2H-chromen-3-yl 3, 4, 5-trihydroxybenzoate, and

(2R, 3R) -7-cyano-5, 5, 8 a-trimethyl-6-oxo-2- (3, 4, 5-trihydroxyphenyl) -3, 4, 4a, 5, 6, 8 a-hexahydro-2H-chromen-3-yl 3, 4, 5-trihydroxybenzoate.

In some embodiments, the present disclosure provides a compound selected from the group consisting of:

(S, E) -3- (4-hydroxystyryl) -3-methyl-6-oxacyclohex-1-enenitrile, and

(3S) -3- (4-hydroxystyryl) -3, 5-dimethyl-6-oxacyclohex-1-enenitrile.

In some embodiments, the present disclosure provides a compound selected from the group consisting of:

(10S, 13S) -4, 4, 10, 13-tetramethyl-3, 17-dioxo-4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17-tetradecahydro-3H-cyclopenta [ a ] phenanthrene-2-carbonitrile,

(10S, 13S, 17S) -17-hydroxy-4, 4, 10, 13-tetramethyl-3-oxo-4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17-tetradecahydro-3H-cyclopenta [ a ] phenanthrene-2-carbonitrile, and

(10S, 13S) -4, 4, 10, 13-tetramethyl-3-oxo-17- (trimethylsilyloxy) -4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17-tetradecahydro-3H-cyclopenta [ a ] phenanthrene-2-carbonitrile.

In some embodiments, the present disclosure provides a compound selected from the group consisting of:

(10S, 13R) -17- (methoxymethyloxy) -10, 13-dimethyl-3-oxo-2, 3, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17-tetradecahydro-1H-cyclopenta [ a ] phenanthrene-4-carbonitrile, and

(10S, 13R) -17-hydroxy-10, 13-dimethyl-3-oxo-2, 3, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17-tetradecahydro-1H-cyclopenteno [ a ] phenanthrene-4-carbonitrile.

In some embodiments, the compounds of the present disclosure are in the form of pharmaceutically acceptable salts. In other embodiments, the compounds of the present disclosure are not in the form of pharmaceutically acceptable salts.

In some embodiments, the compounds of the present disclosure may be esters of the above formula. The ester may, for example, originate from a condensation reaction of the hydroxyl group and the carboxyl group of biotin in the structural formula.

In some embodiments, the compounds of the present disclosure may exist as a mixture of stereoisomers. In other embodiments, the compounds of the present disclosure exist as a single stereoisomer.

In some embodiments, the compounds of the present disclosure may be inhibitors of IFN- γ -induced Nitrous Oxide (NO) production in macrophages, e.g., having an IC of less than 0.2 μ M₅₀The value is obtained.

Other general aspects of the present disclosure contemplate pharmaceutical compositions comprising a compound of the present disclosure as an active ingredient and a pharmaceutically acceptable carrier. The composition may be adapted for administration by, for example, a route selected from: oral, intralipid, intraarterial, intraarticular, intracranial, intradermal, intralesional, intramuscular, intranasal, intraocular, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrarectal, intrathecal, intratracheal, intratumoral, intraumbilical, intravaginal, intravenous, intracapsular, intravitreal, liposomal, topical, transmucosal, oral, parenteral, rectal, subconjunctival, subcutaneous, sublingual, topical, buccal, transdermal, vaginal, in emulsion, in a lipid composition, by catheter, by lavage, by continuous infusion, by perfusion, by inhalation, by injection, by local delivery, by local infusion, by direct bathing of target cells, or any combination thereof. In particular embodiments, the compositions may be formulated for oral delivery. In particular embodiments, the compositions are formulated as hard or soft capsules, tablets, syrups, suspensions, cachets, or elixirs. In certain embodiments, the soft capsule is a gelatin capsule. Certain compositions may comprise a protective coating, such as those formulated for oral delivery. Certain compositions also comprise agents that retard absorption, such as those compositions formulated for oral delivery. Certain compositions may also comprise agents that increase solubility or dispersibility, such as those compositions formulated for oral delivery. Certain compositions may comprise a compound of the present disclosure, wherein the compound is dispersed in liposomes, water and oil emulsions or oil and water emulsions.

Yet another general aspect of the present disclosure contemplates a method of treatment comprising administering to a subject a pharmaceutically effective amount of a compound of the present disclosure. The subject may be, for example, a human. These methods, or any other method of the present disclosure, can further comprise identifying a subject in need of treatment.

Another method of the present disclosure contemplates a method of treating cancer in a subject comprising administering to the subject a pharmaceutically effective amount of a compound of the present disclosure. The cancer may be any type of cancer, such as carcinoma, sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma, or seminoma. Other types of cancer include cancers of the bladder, blood, bone, brain, breast, central nervous system, colon, endometrium, esophagus, genitourinary tract, head, larynx, liver, lung, neck, ovary, pancreas, prostate, spleen, small intestine, large intestine, stomach, or testis. In these methods or any other method, the subject may be a primate. The method or any other method may further comprise identifying a subject in need of treatment. The subject may have a family history or medical history of cancer. In certain embodiments, the subject has symptoms of cancer. The compounds of the invention may be administered by any of the methods described herein, e.g., topically. In certain embodiments, the compound is administered by direct intratumoral injection or by injection into the tumor vasculature. In certain embodiments, the compound may be administered systemically. In certain embodiments, the compound may be administered intravenously, intraarterially, intramuscularly, intraperitoneally, subcutaneously, or orally.

In certain embodiments of the methods for treating cancer in a subject, the method comprises administering to the subject a pharmaceutically effective amount of a compound of the present disclosure, the pharmaceutically effective amount being 0.1-1000 mg/kg. In certain embodiments, the pharmaceutically effective amount is administered as a single dose per day. In certain embodiments, a pharmaceutically effective amount is administered in two or more doses per day. The compounds may be administered, for example, by contacting tumor cells during ex vivo clearance. The method of treatment may include any one or more of the following: a) inducing cytotoxicity in tumor cells; b) killing the tumor cells; c) inducing apoptosis of tumor cells d) inducing differentiation of tumor cells; or e) inhibiting the growth of tumor cells. The tumor cells may be any type of tumor cells, such as leukemia cells. Other types of cells include, for example, bladder cancer cells, breast cancer cells, lung cancer cells, colon cancer cells, prostate cancer cells, liver cancer cells, pancreatic cancer cells, stomach cancer cells, testicular cancer cells, brain cancer cells, ovarian cancer cells, lymphatic cancer cells, skin cancer cells, brain cancer cells, bone cancer cells, or soft tissue cancer cells.

Combination therapy therapies are also contemplated by the present disclosure. For example, with respect to a method of treating cancer in a subject, the method comprising administering to the subject a pharmaceutically effective amount of a compound of the present disclosure, the method may further comprise a treatment selected from the group consisting of: administering a pharmaceutically effective amount of a second drug, radiation therapy, gene therapy, and surgery. Such methods can further comprise (1) contacting the tumor cell with the compound prior to contacting the tumor cell with the second agent, (2) contacting the tumor cell with the second agent prior to contacting the tumor cell with the compound, or (3) simultaneously contacting the tumor cell with the compound and the second agent. In certain embodiments, the second drug can be an antibiotic, an anti-inflammatory drug, an antineoplastic drug, an antiproliferative drug, an antiviral drug, an immunomodulatory drug, or an immunosuppressive drug. The second agent may be an alkylating agent, an androgen receptor modulator, a cytoskeletal disrupting agent, an estrogen receptor modulator, a histone-deacetylase inhibitor, an HMG-CoA reductase inhibitor, a prenyl-protein transferase inhibitor, a retinoid receptor modulator, a topoisomerase inhibitor, or a tyrosine kinase inhibitor. In certain embodiments, the second drug is 5-azacytidine, 5-fluorouracil, 9-cis-retinoic acid, actinomycin D, alitretinoin (alitretinin), all-trans retinoic acid, anamycin, axitinib, belinostat, bevacizumab, bexarotene, bosutin, busulfan, capecitabine, carboplatin, carmustine, CD437, cediranib, cetuximab, chlorambucil, cisplatin, cyclophosphamide, cytarabine, dacarbazine, dasatinib, daunorubicin, decitabine, docetaxel, dolastatin (dolastatin) -10, doxifluridine, doxorubicin, adriamycin, epirubicin, erlotinib, epipodophyllotoxin, gefitinib, gemcitabine, gemtuzumab ozol, omicin, hexamethomsin, idarubicin, ifosfamide, imatinib, and imatinib, Irinotecan, isotretinoin, ixabepilone, lapatinib, LBH589, romotene, mechlorethamine, melphalan, mercaptopurine, methotrexate, mitomycin, mitoxantrone, MS-275, lenatinib, nilotinib, nitrosourea, platinum oxalate, paclitaxel, plicamycin, procarbazine, semaxane, semaphorine, semustine, sodium butyrate, sodium phenylacetate, streptozotocin, suberoylanilide hydroxamic acid, sunitinib, tamoxifen, teniposide, thiotepa (thiopeta), thioguanine, topotecan, TRAIL, trastuzumab, tretinoin, trichostatin A, valproic acid, valrubicin, vandetanib, vinblastine, vincristine, vindesine, or vinorelbine.

Also contemplated are methods of treating or preventing a disease having an inflammatory component in a subject, comprising administering to the subject a pharmaceutically effective amount of a compound of the present disclosure. The disease may be, for example, lupus or rheumatoid arthritis. The disease may be an inflammatory bowel disease, such as crohn's disease or ulcerative colitis. The disease with an inflammatory component may be a cardiovascular disease. The disease having an inflammatory component may be diabetes, such as type 1 or type 2 diabetes. The compounds of the present disclosure may also be useful in the treatment of complications associated with diabetes. Such complications are well known in the art and include, for example, obesity, hypertension, atherosclerosis, coronary heart disease, stroke, peripheral vascular disease, hypertension, nephropathy, neuropathy, muscle necrosis, retinopathy and metabolic syndrome (syndrome X). The disease having an inflammatory component may be a skin disease, such as psoriasis, acne or atopic dermatitis. Administration of the compounds of the present disclosure in methods of treating such skin diseases may be, for example, topical or oral.

The disease having an inflammatory component may be metabolic syndrome (syndrome X). Patients with this syndrome are characterized as having three or more symptoms selected from the following 5 symptoms: (1) abdominal obesity; (2) hypertriglyceridemia; (3) low high density lipoprotein cholesterol (HDL); (4) high blood pressure; and (5) elevated fasting glucose, which may be within the range characteristic of type 2 diabetes if the patient also suffers from diabetes. Each of these symptoms is defined in the Third Report of the National Cholesterol equivalent program Expert Panel on Detection, Evaluation and Treatment of High bloodCholesterol in additives (additive Treatment Panel III, or ATP III), National Institutes of health, 2001, NIH Publication No.01-3670, which is incorporated herein by reference. Patients with metabolic syndrome, whether or not they have or develop overt diabetes, have an increased risk of developing the large and micro-vascular complications listed above that accompany type 2 diabetes, such as atherosclerosis and coronary heart disease.

Another general method of the present disclosure relates to a method of treating or preventing a cardiovascular disease in a subject, comprising administering to the subject a pharmaceutically effective amount of a compound of the present disclosure. The cardiovascular disease may be, for example, atherosclerosis, cardiomyopathy, congenital heart disease, congestive heart failure, myocarditis, rheumatic heart disease, valvular disease, coronary artery disease, endocarditis, or myocardial infarction. Combination therapy is also contemplated for such methods. For example, such methods may further comprise administering a pharmaceutically effective amount of a second drug. The second agent may be, for example, a cholesterol-lowering agent, a hypolipidemic agent, a calcium channel blocker, a hypotensive agent, or an HMG-CoA reductase inhibitor. Non-limiting examples of the second drug include amlodipine, aspirin, ezetimibe, felodipine, lacidipine, lercanidipine, nicardipine, nifedipine, nimodipine, nisoldipine, or nitrendipine. Additional non-limiting examples of the second drug include atenolol, bucindolol, carvedilol, clonidine, doxazosin, indoramine, labetalol, methyldopa, metoprolol, nadolol, oxprenolol, phenoxybenzamine, phentolamine, pindolol, prazosin, propranolol, terazosin, timolol, or tolazoline. The second drug may be, for example, a statin, such as atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, or simvastatin.

Also contemplated are methods of treating or preventing a neurodegenerative disease in a subject comprising administering to the subject a pharmaceutically effective amount of a compound of the present disclosure. The neurodegenerative disease may for example be selected from parkinson's disease, alzheimer's disease, Multiple Sclerosis (MS), huntington's disease and amyotrophic lateral sclerosis. In a particular embodiment, the neurodegenerative disease is alzheimer's disease. In particular embodiments, the neurodegenerative disease is MS, e.g., primary progressive, relapsing-remitting secondary progressive or progressive relapsing MS. The subject can be, for example, a primate. The subject may be a human.

In a particular embodiment of a method of treating or preventing a neurodegenerative disease in a subject, the method comprises administering to the subject a pharmaceutically effective amount of the disclosed compound, the treatment inhibits demyelination of neurons in the brain or spinal cord of the subject. In certain embodiments, the treatment inhibits inflammatory demyelination. In certain embodiments, the treatment inhibits transection of a neuronal axon in the brain or spinal cord of the subject. In certain embodiments, the treatment inhibits neurite transection in the brain or spinal cord of the subject. In certain embodiments, the treatment inhibits neuronal apoptosis in the brain or spinal cord of the subject. In certain embodiments, the treatment stimulates remyelination of neuronal axons in the brain or spinal cord of the subject. In certain embodiments, the treatment restores function lost after onset of MS. In certain embodiments, the treatment prevents the onset of new MS. In certain embodiments, treatment prevents disability resulting from the onset of MS.

One general aspect of the present disclosure contemplates a method of treating or preventing a disorder characterized by overexpression of an iNOS gene in a subject, the method comprising administering to the subject a pharmaceutically effective amount of a compound of the present disclosure.

Another general aspect of the present disclosure contemplates a method of inhibiting IFN- γ -induced nitric oxide production in a cell of a subject, comprising administering to the subject a pharmaceutically effective amount of a compound of the present disclosure.

Yet another general method of the present disclosure contemplates a method of treating or preventing a disorder characterized by overexpression of a COX-2 gene in a subject, comprising administering to the subject a pharmaceutically effective amount of a compound of the present disclosure.

Also contemplated are methods of treating kidney/renal disease (RKD) in a subject comprising administering to the subject a pharmaceutically effective amount of a compound of the disclosure. See U.S. patent application 12/352,473, which is incorporated herein by reference in its entirety. RKD may be derived from, for example, toxic injury. The toxic injury may be derived, for example, from an imaging agent or drug. The drug may be, for example, a chemotherapeutic agent. In certain embodiments, RKD may be derived from ischemia/reperfusion injury. In certain embodiments, RKD is derived from diabetes or hypertension. RKD may be derived from autoimmune diseases. RKD may be further defined as chronic RKD or acute RKD.

In certain methods of treating kidney/renal disease (RKD) in a subject, the method comprises administering a pharmaceutically effective amount of a compound of the present disclosure to a subject who has undergone or is undergoing dialysis. In certain embodiments, the subject has undergone a kidney transplant or is a candidate for undergoing a kidney transplant. The subject may be a primate. The primate can be a human. The subject in this or any other method may be, for example, a cow, horse, dog, cat, pig, mouse, rat, or guinea pig.

The present disclosure also contemplates a method for improving glomerular filtration rate or creatinine clearance in a subject comprising administering to the subject a pharmaceutically effective amount of a compound of the present disclosure.

Methods of synthesis of the disclosed compounds are also contemplated. For example, certain embodiments contemplate methods of making a first compound, wherein the first compound is as follows:

comprising reacting a compound of the formula:

the invention also contemplates a kit, for example a kit comprising: a compound of the present disclosure; and instructions containing information in one or more forms selected from the group consisting of: indicating the disease state of the compound to be administered, stored information on the compound, dosage information, and instructions on how to administer the compound. Kits may comprise a compound of the present disclosure in a multiple dose form.

In some embodiments, the disclosed compounds may be used for the prevention and treatment of diseases and disorders whose pathology involves oxidative stress, inflammation, and dysregulation of inflammatory signaling pathways. In particular embodiments, the compounds of the present invention are useful for treating diseases characterized by overexpression of Inducible Nitric Oxide Synthase (iNOS), inducible cyclooxygenase (COX-2), or both, in affected tissues; high level production of Reactive Oxygen Species (ROS) or Reactive Nitrogen Species (RNS) such as superoxide, hydrogen peroxide, nitric oxide or peroxynitrite salts; or an overproduction of inflammatory cytokines or other inflammation-related proteins such as TNF α, IL-6, IL-1, IL-8, ICAM-1, VCAM-1 and VEGF. In some embodiments, such diseases or disorders may involve undesirable proliferation of certain cells, such as in the case of cancer (e.g., solid tumors, leukemias, myelomas, lymphomas, and other cancers), fibrosis-associated organ failure, or excessive scar hyperplasia. Other such diseases include, but are not limited to, autoimmune diseases such as lupus, rheumatoid arthritis, juvenile onset diabetes, multiple sclerosis, psoriasis and crohn's disease; cardiovascular diseases such as atherosclerosis, heart failure, myocardial infarction, acute coronary syndrome, restenosis following vascular surgery, hypertension and vasculitis; neurodegenerative or neuromuscular diseases such as alzheimer's disease, parkinson's disease, huntington's disease, ALS, and muscular dystrophy; neurological disorders such as epilepsy and dystonia; neuropsychiatric disorders such as major depression, bipolar disorder, post traumatic stress disorder, schizophrenia, anorexia nervosa, ADHD and autism spectrum disorder; retinal diseases such as macular degeneration, diabetic retinopathy, glaucoma, and retinitis; chronic and acute pain syndromes, including inflammatory pain and neuropathic pain; hearing impairment and tinnitus; diabetes and diabetic complications including metabolic syndrome, diabetic nephropathy, diabetic neuropathy and diabetic ulcers; respiratory diseases such as asthma, chronic obstructive pulmonary disease, acute respiratory distress syndrome, and cystic fibrosis; inflammatory bowel disease; osteoporosis, osteoarthritis, and other degenerative diseases of bone and cartilage; acute or chronic organ failure including renal failure, liver failure (including cirrhosis and hepatitis), and pancreatitis; ischemic-reperfusion injury associated with thrombotic or hemorrhagic stroke, subarachnoid hemorrhage, cerebral vasospasm, myocardial infarction, shock, or trauma; organ or tissue transplant complications including acute or chronic transplant failure or rejection and graft versus host disease; skin diseases including atopic dermatitis and acne; sepsis and septic shock; severe inflammation associated with infection, including influenza-associated respiratory inflammation and upper respiratory tract infections; mucositis associated with cancer treatment, including radiation therapy or chemotherapy; and severe burns.

Other objects, features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. Note that simply because a particular compound is assigned to one particular formula does not mean that it cannot be assigned to another formula.

Drawings

The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present disclosure. The invention may be better understood by reference to one of these drawings in combination with the detailed description of specific embodiments presented herein.

Fig. 1-16 and 20-24. Inhibition of NO production. RAW264.7 macrophages were pretreated with DMSO or different concentrations of drug (μ M) for 2 hours and then with IFN γ for 24 hours. Determining the concentration of NO in the culture medium using a Griess reagent system; cell viability was determined using WST-1 reagent.

FIGS. 17-18. Repression of iNOS mRNA induction. RAW264.7 mouse macrophages were pretreated with compound at the indicated concentrations for 2 hours and then stimulated with 10ng/ml IFN γ for an additional 2 hours. mRNA levels of iNOS were quantified by qPCR and shown as values relative to vehicle-treated IFN γ -stimulated samples, which were normalized to a value of 1. Values are the average of PCR reactions in duplicate and with triplicate wells per duplicate.

Fig. 19. iNOS Western blot in RAW264.7 mouse macrophages. RAW264.7 cells were pretreated with the indicated compounds for 2 hours and then stimulated with 10ng/ml IFN γ for an additional 24 hours. Levels of iNOS protein were determined by immunoblotting. Actin was used as loading control.

Description of illustrative embodiments

Disclosed herein are novel compounds, e.g., having antioxidant and anti-inflammatory properties, methods of making the same, and methods of using the same, including methods for treating and/or preventing diseases.

I. Definition of

"basic skeleton" is defined as: (1) atoms and sigma-bonds forming the backbone of each of the two rings of the organic compound (e.g., a natural or non-natural product), the two rings being furthest apart from each other when the natural product is drawn in a two-dimensional structure, provided that the two rings are connected to each other via a backbone that is entirely composed of carbon-carbon bonds (e.g., single, double, triple, aromatic or non-aromatic bonds); and (2) an atom and a sigma-bond linking any of the backbones of the two rings defined in (1).

As used herein, "hydrogen" means-H; "hydroxy" means-OH; "oxygen" means ═ O; "halo" means independently-F, -Cl, -Br, or-I; "amino" means-NH ₂(see below for a definition of groups comprising the term amino, e.g. alkylamino); "hydroxyamino" means-NHOH; "nitro" means-NO₂(ii) a Imino means ═ NH (see below for the definition of groups containing the term imino, e.g. alkylamino); "cyano" means-CN; "azido" means-N₃(ii) a "mercapto" means-SH; "sulfur" means ═ S; "sulfonamido" means-NHS (O)₂- (see below for definition of groups comprising the term sulfonamide group, e.g. alkylsulfonamide group); "Sulfonyl" means-S (O)₂- (see below for definition of groups comprising the term sulfonyl, e.g. alkylsulfonyl); and "silyl" means-SiH₃(see below for a definition of groups comprising the term silyl, e.g. alkylsilyl).

The following parenthetical subscripts are further defined for the following groups: "(Cn)" is defined as the exact number of carbon atoms in the group (n). "(C.ltoreq.n)" is defined as the maximum number of carbon atoms (n) that can be present in a group, in this way the minimum number of carbon atoms is at least one, but in addition may be as small a number as possible for the group in question. For example, the group "alkenyl" is to be understood _(C≤8)"the minimum number of carbon atoms is 2. For example, "alkoxy group_(C≤10)Those alkoxy groups having from 1 to 10 carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or any range derivable therein (e.g., 3-10 carbon atoms)) are named. (Cn-n ') defines the minimum (n) and maximum (n') values of carbon atoms in the group. Similarly, "alkyl group_(C2-10)Those alkyl groups having from 2 to 10 carbon atoms (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, or any range derivable therein (e.g., 3-10 carbon atoms)) are named.

The term "alkyl", when used in the modification without "substitution", refers to a non-aromatic monovalent radical having a saturated carbon atom as the point of attachment, which is a linear or branched, cyclic or acyclic structure, no carbon-carbon double or triple bonds, and no other atoms besides carbon and hydrogen. group-CH₃(Me)、-CH₂CH₃(Et)、-CH₂CH₂CH₃(n-Pr)、-CH(CH₃)₂(iso-Pr)、-CH(CH₂)₂(cyclopropyl), -CH₂CH₂CH₂CH₃(n-Bu)、-CH(CH₃)CH₂CH₃(sec-butyl), -CH₂CH(CH₃)₂(isobutyl), -C (CH)₃)₃(tert-butyl), -CH₂C(CH₃)₃(neopentyl), cyclobutyl, cyclopentyl, cyclohexyl and cyclohexylmethyl are non-limiting examples of alkyl groups. The term "substituted alkyl" refers to a non-aromatic monovalent radical having a saturated carbon atom as the point of attachment, which is linear or branched, cyclic or acyclic, free of carbon-carbon double or triple bonds, and at least one Each atom is independently selected from N, O, F, Cl, Br, I, Si, P and S. The following groups are non-limiting examples of substituted alkyls: -CH₂OH、-CH₂Cl、-CH₂Br、-CH₂SH、-CF₃、-CH₂CN、-CH₂C(O)H、-CH₂C(O)OH、-CH₂C(O)OCH₃、-CH₂C(O)NH₂、-CH₂C(O)NHCH₃、-CH₂C(O)CH₃、-CH₂OCH₃、-CH₂OCH₂CF₃、-CH₂OC(O)CH₃、-CH₂NH₂、-CH₂NHCH₃、-CH₂N(CH₃)₂、-CH₂CH₂Cl、-CH₂CH₂OH、-CH₂CF₃、-CH₂CH₂OC(O)CH₃、-CH₂CH₂NHCO₂C(CH₃)₃and-CH₂Si(CH₃)₃。

The term "alkanediyl", when used without the modifier "substituted", refers to a non-aromatic divalent radical wherein alkanediyl is linked in two sigma-bonds, has one or two saturated carbon atoms as the point of attachment, is a linear or branched, cyclic or acyclic structure, has no carbon-carbon double or triple bonds, and has no atoms other than carbon and hydrogen. group-CH₂- (methylene), -CH₂CH₂-、-CH₂C(CH₃)₂CH₂-、-CH₂CH₂CH₂-andare non-limiting examples of alkanediyl. The term "substituted alkanediyl" refers to a non-aromatic monovalent group in which an alkynediyl group is connected in two sigma-bonds, has as a point of connection one or two saturated carbon atoms, which is a linear or branched, cyclic or acyclic structure, free of carbon-carbon double or triple bonds, and at least one atom is independently selected from N, O, F, Cl, Br, I, Si, P, and S. The following groups areNon-limiting examples of substituted alkanediyl groups: -CH (F) -, -CF₂-、-CH(Cl)-、-CH(OH)-、-CH(OCH₃) -and-CH₂CH(Cl)-。

The term "alkenyl", when used without the modifier "substituted", refers to a monovalent group having a non-aromatic carbon atom as the point of attachment, either a linear or branched, cyclic or acyclic structure, at least one non-aromatic carbon-carbon double bond, no carbon-carbon triple bonds, and no atoms other than carbon and hydrogen. Non-limiting examples of alkenyl groups include: -CH ═ CH ₂(vinyl), -CH ═ CHCH₃、-CH＝CHCH₂CH₃、-CH₂CH＝CH₂(allyl), -CH₂CH＝CHCH₃and-CH ═ CH-C₆H₅. The term "substituted alkenyl" refers to a monovalent group having a non-aromatic carbon atom as the point of attachment, having at least one non-aromatic carbon-carbon double bond, no carbon-carbon triple bond, linear or branched, cyclic, or acyclic structure, and at least one atom is independently selected from the group consisting of N, O, F, Cl, Br, I, Si, P, and S. The radicals-CH ═ CHF, -CH ═ CHCl and-CH ═ CHBr are non-limiting examples of substituted alkenyl groups.

The term "alkenediyl", when used without the modifier "substituted", refers to a non-aromatic divalent radical in which the alkenediyl is linked in two sigma-bonds, has two carbon atoms as points of attachment, is a linear or branched, cyclic, or acyclic structure, has at least one non-aromatic carbon-carbon double bond, no carbon-carbon triple bonds, and no atoms other than carbon and hydrogen. The radicals-CH-, -CH-C (CH)₃)CH₂-、-CH＝CHCH₂-andare non-limiting examples of alkenediyl groups. The term "substituted enediyl" refers to a non-aromatic divalent radical in which the enediyl radical is linked in two sigma-bonds, has two carbon atoms as points of attachment, is a linear or branched, cyclic or acyclic structure, has at least one non-aromatic carbon-carbon double bond, has no carbon-carbon triple bonds, and At least one atom is independently selected from N, O, F, Cl, Br, I, Si, P and S. The following groups are non-limiting examples of substituted alkenediyl groups: -CF ═ CH-, -c (oh) ═ CH-, and-CH₂CH＝C(Cl)-。

The term "alkynyl", when used without the modifier "substituted", refers to a monovalent group having a non-aromatic carbon atom as the point of attachment, having a linear or branched, cyclic or acyclic structure, at least one carbon-carbon triple bond, and no other atoms besides carbon and hydrogen. The group-C.ident.CH, -C.ident.CCH₃、-C≡CC₆H₅and-CH₂C≡CCH₃Are non-limiting examples of alkynyl groups. The term "substituted alkynyl" refers to a monovalent group having a non-aromatic carbon atom as the point of attachment, and having at least one carbon-carbon triple bond, being a linear or branched, cyclic or acyclic structure, and at least one atom being independently selected from the group consisting of N, O, F, Cl, Br, I, Si, P, and S. The group-C ≡ CSi (CH)₃)₃Are non-limiting examples of substituted alkynyl groups.

The term "alkynediyl", when used without the modifier "substituted", refers to a non-aromatic divalent radical in which the alkynediyl is linked by two sigma-bonds, has two carbon atoms as points of attachment, has a linear or branched, cyclic or acyclic structure, at least one carbon-carbon triple bond, and is free of atoms other than carbon and hydrogen. The groups-C.ident.C-, -C.ident.CCH ₂-and-C ≡ CCH (CH)₃) -is a non-limiting example of an alkyndiyl group. The term "substituted alkynediyl" refers to a non-aromatic divalent radical in which the alkynediyl is linked in two sigma-bonds, has two carbon atoms as points of attachment, has a linear or branched, cyclic or acyclic structure, at least one carbon-carbon triple bond, and at least one atom is independently selected from the group consisting of N, O, F, Cl, Br, I, Si, P, and S. The groups-C.ident.CCFH-and-C.ident.CHCH (Cl) -are non-limiting examples of substituted alkynediyl groups.

The term "aryl", when used without the modifier "substituted", refers to a monovalent group having an aromatic carbon atom as the point of attachment, andthe carbon atoms form part of a six-membered aromatic ring structure in which the ring atoms are all carbon and in which the monovalent radical does not contain atoms other than carbon and hydrogen. Non-limiting examples of aryl groups include phenyl (Ph), methylphenyl, (dimethyl) phenyl, -C₆H₄CH₂CH₃(ethylphenyl), -C₆H₄CH₂CH₂CH₃(propylphenyl), -C₆H₄CH(CH₃)₂、-C₆H₄CH(CH₂)₂、-C₆H₃(CH₃)CH₂CH₃(methylethylphenyl), -C₆H₄CH＝CH₂(vinylphenyl), -C₆H₄CH＝CHCH₃、-C₆H₄C≡CH、-C₆H₄C≡CCH₃Naphthyl and monovalent radicals derived from biphenyl. The term "substituted aryl" refers to a monovalent group having, as a point of attachment, an aromatic carbon atom that forms part of a six-membered aromatic ring structure, wherein the ring atoms are all carbon, and wherein the monovalent group further has at least one atom independently selected from the group consisting of N, O, F, Cl, Br, I, Si, P, and S. Non-limiting examples of substituted aryl groups include the groups: -C ₆H₄F、-C₆H₄Cl、-C₆H₄Br、-C₆H₄I、-C₆H₄OH、-C₆H₄OCH₃、-C₆H₄OCH₂CH₃、-C₆H₄OC(O)CH₃、-C₆H₄NH₂、-C₆H₄NHCH₃、-C₆H₄N(CH₃)₂、-C₆H₄CH₂OH、-C₆H₄CH₂OC(O)CH₃、-C₆H₄CH₂NH₂、-C₆H₄CF₃、-C₆H₄CN、-C₆H₄CHO、-C₆H₄CHO、-C₆H₄C(O)CH₃、-C₆H₄C(O)C₆H₅、-C₆H₄CO₂H、-C₆H₄CO₂CH₃、-C₆H₄CONH₂、-C₆H₄CONHCH₃and-C₆H₄CON(CH₃)₂。

The term "aryldiyl (aryldiyl), when used without the modifier" substituted ", refers to a divalent group wherein the aryldiyl is connected in two sigma-bonds, having as a point of attachment two aromatic carbon atoms that form part of one or more six-membered aromatic ring structures wherein the ring atoms are all carbon, and wherein the monovalent group contains no atoms other than carbon and hydrogen. Non-limiting examples of aryldiyl groups include:

the term "substituted aryldiyl" refers to a divalent group wherein the aryldiyl is connected in two sigma-bonds, having as a point of attachment two aromatic carbon atoms that form part of one or more six-membered aromatic ring structures wherein the ring atoms are all carbon, and wherein the divalent group further has at least one atom independently selected from the group consisting of N, O, F, Cl, Br, I, Si, P, and S.

The term "aralkyl", when used without the modifier "substituted", refers to a monovalent radical-alkanediyl-aryl, wherein the terms alkanediyl and aryl are each used in a manner consistent with the definition provided above. Non-limiting examples of aralkyl groups are: phenylmethyl (benzyl, Bn), 1-phenyl-ethyl, 2-phenyl-ethyl, indenyl and 2, 3-dihydro-indenyl, with the proviso that indenyl and 2, 3-dihydro-indenyl are the only examples of aralkyl groups at present whose point of attachment is in each case one of the saturated carbon atoms. When the terms "aralkyl", "substituted" and "substituted" modifiers are used together, either or both of the alkanediyl and aryl groups are substituted. Non-limiting examples of substituted aralkyl groups are: (3-chlorophenyl) -methyl, 2-oxo-2-phenyl-ethyl (phenylcarbonylmethyl), 2-chloro-2-phenyl-ethyl, chromanyl wherein the point of attachment is one of the saturated carbon atoms, and tetrahydroquinolinyl wherein the point of attachment is one of the saturated atoms.

The term "heteroaryl", when used without the modifier "substituted", refers to a monovalent group having, as a point of attachment, an aromatic carbon or nitrogen atom that forms part of an aromatic ring structure in which at least one of the ring atoms is nitrogen, oxygen, or sulfur, and in which the monovalent group contains no atoms other than carbon, hydrogen, aromatic nitrogen, aromatic oxygen, and aromatic sulfur. Non-limiting examples of aryl groups include acridinyl, furyl, imidazoimidazolyl, imidazopyrazolyl, imidazopyridyl, imidazopyrimidinyl, indolyl, imidazolinyl, methylpyridinyl, and the like,Azolyl, phenylimidazolyl, pyridyl, pyrrolyl, pyrimidinyl, pyrazinyl, quinolinyl, quinazolinyl, quinoxalinyl, tetrahydroquinolinyl, thienyl, triazinyl, pyrrolopyridyl, pyrrolopyrimidinyl, pyrrolopyrazinyl, pyrrolotriazinyl, pyrroloimidazolyl, benzopyranyl (where the point of attachment is one of the aromatic atoms), and chromanyl (where the point of attachment is one of the aromatic atoms). The term "substituted heteroaryl" refers to a monovalent group having an aromatic carbon or nitrogen atom as the point of attachment, said carbon or nitrogen atom forming part of an aromatic ring structure, wherein at least one of the ring atoms is nitrogen, oxygen, or sulfur, and wherein the monovalent group further has at least one atom independently selected from the group consisting of non-aromatic nitrogen, non-aromatic oxygen, non-aromatic sulfur, F, Cl, Br, I, Si, and P.

The term "heteroaryldiyl," when used without the modifier "substituted," refers to a divalent group wherein the heteroaryldiyl group is joined in two sigma-bonds with an aromatic carbon or nitrogen atom as the point of attachment, the carbon or nitrogen atom having two aromatic atoms as the point of attachment, the carbon atom forming part of one or more six-membered aromatic ring structures wherein the ring atoms are all carbon, and wherein the monovalent group contains no atoms other than carbon and hydrogen. Non-limiting examples of heteroaryl diradicals include:

the term "substituted heteroaryldiyl" refers to a divalent group wherein the heteroaryldiyl group is connected by two sigma-bonds, having as a point of attachment two aromatic carbon atoms that form part of one or more six-membered aromatic ring structures wherein the ring atoms are all carbon, and wherein the divalent group further has at least one atom independently selected from the group consisting of N, O, F, Cl, Br, I, Si, P, and S.

The term "heteroaralkyl", when used without the modifier "substituted", refers to a monovalent radical-alkanediyl-heteroaryl, wherein the terms alkanediyl and heteroaryl are each used in a manner consistent with the definition provided above. Non-limiting examples of aralkyl groups are: pyridylmethyl and thienylmethyl. When the term "heteroaralkyl" is used in conjunction with the "substituted" modifier, either or both of alkanediyl and heteroaryl are substituted.

The term "acyl", when used without the modifier "substituted", refers to a monovalent group having a carbonyl carbon atom as the point of attachment, and also having a linear or branched, cyclic or acyclic structure, with no atoms other than carbon or hydrogen other than the oxygen atom of the carbonyl group. The group-CHO, -C (O) CH₃(acetyl, Ac), -C (O) CH₂CH₃、-C(O)CH₂CH₂CH₃、-C(O)CH(CH₃)₂、-C(O)CH(CH₂)₂、-C(O)C₆H₅、-C(O)C₆H₄CH₃、-C(O)C₆H₄CH₂CH₃、-COC₆H₃(CH₃)₂and-C (O) CH₂C₆H₅Are non-limiting examples of acyl groups. Thus, the term "acyl" embraces,but are not limited to, groups that are often referred to as "alkylcarbonyl" and "arylcarbonyl". The term "substituted acyl" refers to a monovalent group having a carbonyl carbon atom as the point of attachment, and also having a linear or branched, cyclic or acyclic structure, in addition to the carbonyl oxygen, having at least one atom independently selected from the group consisting of N, O, F, Cl, Br, I, Si, P, and S. group-C (O) CH₂CF₃、-CO₂H (carboxyl), -CO₂CH₃(methyl carboxyl), -CO₂CH₂CH₃、-CO₂CH₂CH₂CH₃、-CO₂C₆H₅、-CO₂CH(CH₃)₂、-CO₂CH(CH₂)₂、-C(O)NH₂(carbamoyl), -C (O) NHCH₃、-C(O)NHCH₂CH₃、-CONHCH(CH₃)₂、-CONHCH(CH₂)₂、-CON(CH₃)₂、-CONHCH₂CF₃-CO-pyridyl, -CO-imidazolyl and-C (O) N₃Are non-limiting examples of substituted acyl groups. The term "substituted acyl" includes, but is not limited to, "heteroarylcarbonyl".

The term "alkylene", when used without the modifier "substituted", refers to a divalent radical ═ CRR ', where the alkylene is connected by one sigma-bond and one pi-bond, where R and R ' are independently hydrogen, alkyl, or R and R ' together represent an alkanediyl group. Non-limiting examples of alkylene groups include: CH (CH) ₂、＝CH(CH₂CH₃) And ═ C (CH)₃)₂. The term "substituted alkylene" refers to the group ═ CRR ', where the alkylene is connected by one sigma-bond and one pi-bond, where R and R ' are independently hydrogen, alkyl, substituted alkyl, or R and R ' together represent a substituted alkanediyl, provided that either R or R ' is a substituted alkyl or R and R ' together represent a substituted alkanediyl.

The term "alkoxy", when used without the modifier "substituted", refers to the group-OR, wherein R is alkyl, as defined above for the term. Non-limiting examples of alkoxy groups includeComprises the following steps: -OCH₃、-OCH₂CH₃、-OCH₂CH₂CH₃、-OCH(CH₃)₂、-OCH(CH₂)₂-O-cyclopentyl and-O-cyclohexyl. The term "substituted alkoxy" refers to the group-OR, where R is substituted alkyl, as defined above for the term. For example, -OCH₂CF₃Is a substituted alkoxy group.

Similarly, the terms "alkenyloxy", "alkynyloxy", "aryloxy", "aralkoxy", "heteroaryloxy", "heteroarylalkoxy", and "acyloxy", when used without the modifier "substituted", refer to groups defined as-OR, wherein R is independently alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and acyl, as defined above for those terms. When any of the terms alkenyloxy, alkynyloxy, aryloxy, aralkyloxy, and acyloxy are modified by "substitution," it refers to the group-OR, where R is substituted alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and acyl, respectively.

The term "alkylamino", when not used in conjunction with the modifier "substituted", refers to the group-NHR, where R is alkyl, as defined above for that term. Non-limiting examples of alkylamino groups include: -NHCH₃、-NHCH₂CH₃、-NHCH₂CH₂CH₃、-NHCH(CH₃)₂、-NHCH(CH₂)₂、-NHCH₂CH₂CH₂CH₃、-NHCH(CH₃)CH₂CH₃、-NHCH₂CH(CH₃)₂、-NHC(CH₃)₃-NH-cyclopentyl and-NH-cyclohexyl. The term "substituted alkylamino" refers to the group-NHR, where R is substituted alkyl, as defined above for the term. For example, -NHCH₂CF₃Is a substituted alkylamino group.

The term "dialkylamino", when used without the modifier "substituted", refers to the group-NRR ', where R and R' may be the same or different alkyl groups, or R and RR' may together represent an alkanediyl group having two or more saturated carbon atoms wherein at least two saturated carbon atoms are attached to a nitrogen atom. Non-limiting examples of dialkylamino groups include: -NHC (CH)₃)₃、-N(CH₃)CH₂CH₃、-N(CH₂CH₃)₂N-pyrrolidinyl and N-piperidinyl. The term "substituted dialkylamino" refers to the group-NRR ', where R and R can be the same or different substituted alkyl groups, one of R or R ' is an alkyl group and the other is a substituted alkyl group, or R and R ' can together represent a substituted alkanediyl group having two or more saturated carbon atoms wherein at least two saturated carbon atoms are attached to a nitrogen atom.

The terms "alkoxyamino", "alkenylamino", "alkynylamino", "arylamino", "aralkylamino", "heteroarylamino", "heteroaralkylamino", and "alkylsulfonylamino", when used without the modifier "substituted", refer to groups defined as — NHR, wherein R is alkoxy, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and alkylsulfonyl, respectively, as defined above for those terms. A non-limiting example of an arylamino group is-NHC₆H₅. When any of the terms alkoxyamino, alkenylamino, alkynylamino, arylamino, aralkylamino, heteroarylamino, heteroaralkylamino, and alkylsulfonylamino are modified by "substitution", it refers to the group-NHR, where R is substituted alkoxy, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and alkylsulfonyl, respectively.

The term "acylamino" (acylamino), when used without the modifier "substituted", refers to the group-NHR, wherein R is acyl, as defined above for the term. A non-limiting example of an acylamino group is-NHC (O) CH₃. When the term amido is used in conjunction with the "substituted" modifier, it refers to a group defined as-NHR, where R is a substituted acyl group, as defined above for the term. The group-NHC (O) OCH ₃And NHC (O) NHCH₃Is a non-limiting group of substituted amideIllustrative examples.

The term "alkylimino", when used without the modifier "substituted", refers to the group NR, wherein alkylimino is connected by one σ -bond and one pi-bond, wherein R is alkyl, as defined above for the term. Non-limiting examples of alkylimino groups include: as NCH₃、＝NCH₂CH₃And ═ N-cyclohexyl. The term "substituted alkylimino" refers to the group NR, wherein alkylimino is linked by one σ -bond and one pi-bond, wherein R is substituted alkyl, as defined above for the term. E.g., ═ NCH₂CF₃Is a substituted alkylimino group.

Similarly, the terms "alkenylimino", "alkynylimino", "arylimino", "aralkylimino", "heteroarylimino", "heteroaralkylimino", and "acylimino", when used without the modifier "substituted", refer to a group defined as NR, wherein the alkylimino group is connected by one σ -bond and one pi-bond, wherein R is alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and acyl, respectively, as defined above for those terms. When any of the terms alkenylimino, alkynylimino, arylimino, aralkylimino, and acylimino is modified by "substituted," it refers to a group where the alkylimino is connected by one sigma-bond and one pi-bond, where R is a substituted alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and acyl group, respectively.

The term "fluoroalkyl", when used without the modifier "substituted", refers to an alkyl group, as defined above for the term, in which one or more fluorine has been replaced with hydrogen. group-CH₂F、-CF₃and-CH₂CF₃Are non-limiting examples of fluoroalkyl groups. The term "substituted fluoroalkyl" refers to a non-aromatic monovalent radical having a saturated carbon atom as the point of attachment, having a linear or branched, cyclic or acyclic structure, at least one fluorine atom, no carbon-carbon double or triple bonds,and at least one atom is independently selected from N, O, Cl, Br, I, Si, P and S. The following groups are non-limiting examples of substituted fluoroalkyl groups: -CFHOH.

The term "alkylthio", when used without the modifier "substituted", refers to the group-SR, wherein R is alkyl, as defined above for the term. Non-limiting examples of alkylthio groups include: -SCH₃、-SCH₂CH₃、-SCH₂CH₂CH₃、-SCH(CH₃)₂、-SCH(CH₂)₂-S-cyclopentyl and-S-cyclohexyl. The term "substituted alkylthio" refers to the group-SR, wherein R is substituted alkyl, as defined above for the term. For example, -SCH₂CF₃Is a substituted alkylthio group.

Similarly, the terms "alkenylthio", "alkynylthio", "arylthio", "aralkylthio", "heteroarylthio", "heteroarylalkylthio" and "acylthio", when used without the modifier "substituted", refer to the group defined as-SR, wherein R is alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl and acyl, respectively, as defined above for those terms. When any of the terms alkenylthio, alkynylthio, arylthio, aralkylthio, heteroarylthio, heteroaralkylthio, and acylthio are modified by "substitution", it refers to the group-SR, wherein R is substituted alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and acyl, respectively.

The term "thioacyl", when used without the modifier "substituted", refers to a monovalent radical having the carbon atom of the thiocarbonyl group as the point of attachment, and also having a linear or branched, cyclic or acyclic structure, with no additional atoms other than carbon or hydrogen other than the carbonyl sulfide atom. The group-CHS, -C (S) CH₃、-C(S)CH₂CH₃、-C(S)CH₂CH₂CH₃、-C(S)CH(CH₃)₂、-C(S)CH(CH₂)₂、-C(S)C₆H₅、-C(S)C₆H₄CH₃、-C(S)C₆H₄CH₂CH₃、-C(S)C₆H₃(CH₃)₂and-C (S) CH₂C₆H₅Are non-limiting examples of thioacyl groups. The term "thioacyl" thus includes, but is not limited to, the groups that are often referred to as "alkylthiocarbonyl" and "arylthiocarbonyl". The term "substituted thioacyl" refers to a radical having as a point of attachment a carbon atom that is part of a thiocarbonyl group, and also having a linear or branched, cyclic or acyclic structure, and, in addition to a carbonylthio atom, at least one atom independently selected from the group consisting of N, O, F, Cl, Br, I, Si, P, and S. The group-C (S) CH₂CF₃、-C(S)O₂H、-C(S)OCH₃、-C(S)OCH₂CH₃、-C(S)OCH₂CH₂CH₃、-C(S)OC₆H₅、-C(S)OCH(CH₃)₂、-C(S)OCH(CH₂)₂、-C(S)NH₂and-C (S) NHCH₃Are non-limiting examples of substituted thioacyl groups. The term "substituted thioacyl" includes, but is not limited to, "heteroarylthiocarbonyl".

The term "alkylsulfonyl", when used without the modifier "substituted", refers to the group-S (O) ₂R, wherein R is alkyl, as defined above for the term. Non-limiting examples of alkylsulfonyl groups include: -S (O)₂CH₃、-S(O)₂CH₂CH₃、-S(O)₂CH₂CH₂CH₃、-S(O)₂CH(CH₃)₂、-S(O)₂CH(CH₂)₂、-S(O)₂-cyclopentyl and-S (O)₂-cyclohexyl. The term "substituted alkylsulfonyl" refers to the group-S (O)₂R, wherein R is substituted alkyl, as defined above for the term. For example, -S (O)₂CH₂CF₃Is a substituted alkylsulfonyl group.

Similarly, the terms "alkenylsulfonyl", "alkynylsulfonyl", "arylsulfonyl", "aralkylsulfonyl"),"Heteroarylsulfonyl" and "heteroaralkylsulfonyl", when used without the modifier "substituted", is defined as-S (O)₂The group of R, wherein R is independently alkenyl, alkynyl, aryl, aralkyl, heteroaryl, and heteroaralkyl, as defined above for those terms. When any of the terms alkenylsulfonyl, alkynylsulfonyl, arylsulfonyl, aralkylsulfonyl, heteroarylsulfonyl and heteroaralkylsulfonyl are modified by "substitution", it refers to the group-S (O)₂R, wherein R is independently substituted alkenyl, alkynyl, aryl, aralkyl, heteroaryl, and heteroaralkyl.

The term "alkylammonium," when used without the modifier "substituted," is defined as-NH ₂R⁺、-NHRR′⁺or-NRR' R⁺Wherein R, R 'and R "are the same or different alkyl groups, or any combination of any two of R, R' and R" may together represent an alkanediyl group. Non-limiting examples of alkylammonium cations include: -NH₂(CH₃)⁺、-NH₂(CH₂CH₃)+、-NH₂(CH₂CH₂CH₃)+、-NH(CH₃)₂ ⁺、-NH(CH₂CH₃)₂ ⁺、-NH(CH₂CH₂CH₃)₂ ⁺、-N(CH₃)₃ ⁺、-N(CH₃)(CH₂CH₃)₂ ⁺、-N(CH₃)₂(CH₂CH₃)⁺、-NH₂C(CH₃)₃ ⁺-NH (cyclopentyl)₂ ⁺and-NH₂(cyclohexyl group)⁺. The term "substituted alkylammonium" refers to-NH₂R⁺、-NHRR′⁺or-NRR' R⁺Wherein at least one of R, R 'and R' is a substituted alkyl group or two of R, R 'and R' may together represent a substituted alkanediyl group. When more than one of R, R 'and R' are substituted alkyl, they may be the same or different. R, R 'and R' being neither a substituted alkyl group nor a substituted alkanediyl groupOne is either alkyl, either the same or different, or may together represent an alkanediyl group having two or more carbon atoms, wherein at least two carbon atoms are bonded to the nitrogen atom shown in the formula.

The term "alkylsulfonium", when not used in conjunction with the modifier "substituted", refers to the group-SRR'⁺Wherein R and R 'may be the same or different alkyl groups, or R and R' may together represent an alkanediyl group. Non-limiting examples of alkyl sulfonium groups include: -SH (CH) ₃)⁺、-SH(CH₂CH₃)⁺、-SH(CH₂CH₂CH₃)⁺、-S(CH₃)₂ ⁺、-S(CH₂CH₃)₂ ⁺、-S(CH₂CH₂CH₃)₂ ⁺-SH (cyclopentyl)⁺and-SH (cyclohexyl)⁺. The term "substituted alkylsulfonium" refers to the group-SRR'⁺Wherein R and R 'may be the same or different substituted alkyl, one of R or R' is alkyl and the other is substituted alkyl, or R and R may together represent substituted alkanediyl. For example, -SH (CH)₂CF₃)⁺Is a substituted alkyl sulfonium group.

The term "alkylsilyl", when used without the modifier "substituted", means defined as-SiH₂A monovalent radical of R, -SiHRR ' or-SiRR ' R ', where R, R ' and R ' may be the same or different alkyl groups, or any combination of two of R, R ' and R ' may together represent an alkanediyl group. radical-SiH₂CH₃、-SiH(CH₃)₂、-Si(CH₃)₃and-Si (CH)₃)₂C(CH₃)₃Non-limiting examples of unsubstituted alkylsilyl groups. The term "substituted alkylsilyl" refers to-SiH₂R, -SiHRR ' or-SiRR ' R ', wherein at least one of R, R ' and R ' is a substituted alkyl or two of R, R ' and R ' may together represent a substituted alkanediyl group. When more than one of R, R 'and R' are substituted alkyl, they may be the same or different. Is neither to getAny of R, R 'and R' where alkyl is also not a substituted alkanediyl may be alkyl, either the same or different, or may together represent an alkanediyl group having two or more saturated carbon atoms wherein at least two carbon atoms are attached to a silicon atom.

Further, the atoms comprising the compounds of the present disclosure are intended to include all isotopic forms of such atoms. Isotopes, as used herein, include those atoms having the same number of atoms but different mass numbers. By way of general example and not limitation, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include¹³C and¹⁴C. similarly, it is contemplated that one or more than one carbon atom of the disclosed compounds may be replaced by one or more silicon atoms. Furthermore, it is contemplated that one or more than one oxygen atom of the disclosed compounds may be replaced by one or more sulfur or selenium atoms.

Compounds having a formula represented by a dashed chemical bond are intended to include chemical formulas that optionally have zero, one, or more than one double bond. Thus, for example, the structureComprises a structure

As will be understood by those skilled in the art, none of such ring atoms form part of more than one double bond.

Any undefined valency on an atom of the structure shown in this application implicitly represents a hydrogen atom bonded to the atom.

Showing the Ring Structure with an unconnected "R" group indicates that at this RingAny of the hydrogen-containing atoms defined above may be substituted with an R group. In the case of a divalent R group (e.g., oxo, imino, thio, alkylene, etc.), any pair of implied hydrogen atoms attached to one atom of the ring may be substituted with the R group. This concept is exemplified as follows: RepresentsOr

As used herein, "chiral auxiliary" refers to a removable chiral group capable of affecting the stereoselectivity of a reaction. Those skilled in the art are familiar with such compounds, and many such compounds are commercially available.

The words "a" or "an" when used in conjunction with the term "comprising" in the claims and/or the specification mean "a", but also have the meaning "one or more", "at least one", and "one or more".

Throughout this application, the term "about" is used to refer to a value that includes variations in the inherent error of the instrument, the method used to determine the value, or variations that exist in the study subject.

The terms "comprising," "having," and "including" are open-ended linking verbs. Any form or tense of one or more of these verbs, such as "comprising," "including," "having," "including," and "including," is open-ended. For example, any method that "comprises," "has," or "has" one or more steps is not limited to possessing only those one or more steps, and also encompasses other unlisted steps.

The term "effective" as used in the specification and/or claims means sufficient to achieve a desired, expected, or expected result.

The term "hydrate" when used as a modifier of a compound means that the compound has less than one (e.g., hemihydrate), one (e.g., monohydrate), or more than one (e.g., dihydrate) water molecule associated with each chemical molecule, e.g., the compound in solid form.

As used herein, the term "IC₅₀"refers to the amount of inhibitor that achieves 50% of maximal response.

An "isomer" of a first compound is a separate compound in which each molecule contains the same constituent atoms as the first compound, but the configuration of these atoms in three-dimensional space is different.

As used herein, the term "patient" or "subject" refers to a living mammalian organism, such as a human, monkey, cow, sheep, goat, dog, cat, mouse, rat, guinea pig, or transgenic species thereof. In certain embodiments, the patient or subject is a primate. Non-limiting examples of human subjects are adults, adolescents, infants and fetuses.

By "pharmaceutically acceptable" it is meant that it is useful in preparing pharmaceutical compositions that are generally safe, non-toxic, biologically and otherwise undesirable, and include those acceptable for veterinary use as well as human pharmaceutical use.

By "pharmaceutically acceptable salt" is meant a salt of the disclosed compound which is pharmaceutically acceptable as defined above and which has the desired pharmacological activity. Such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like; or with organic acids such as 1, 2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, 2-naphthalenesulfonic acid, 3-phenylpropionic acid, 4' -methylenebis (3-hydroxy-2-ene-1-carboxylic acid), 4-methylbicyclo [2.2.2] oct-2-ene-1-carboxylic acid, acetic acid, aliphatic monocarboxylic and dicarboxylic acids, aliphatic sulfuric acid, aromatic sulfuric acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carbonic acid, cinnamic acid, citric acid, cyclopentylpropionic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, heptanoic acid, hexanoic acid, hydroxynaphthoic acid, lactic acid, dodecylsulfuric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid, o- (4-hydroxybenzoyl) benzoic acid, Oxalic acid, p-chlorobenzenesulfonic acid, phenyl-substituted alkanoic acid, propionic acid, p-methylbenzenesulfonic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, tartaric acid, t-butylacetic acid, trimethylacetic acid, and the like. Pharmaceutically acceptable salts also include base addition salts, which may be formed when an acid proton present is capable of reacting with an inorganic or organic base. Acceptable inorganic bases include sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide, and calcium hydroxide. Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like. It will be appreciated that the particular anion or cation forming part of any salt of the invention is not critical, so long as the salt as a whole is pharmaceutically acceptable. Additional examples of pharmaceutically acceptable Salts and methods of making and using the same are found in the Handbook of Pharmaceutical Salts: properties, and Use (2002).

As used herein, "predominantly one enantiomer" means that the compound contains at least about 85% of one enantiomer, or more preferably at least about 90% of one enantiomer, or even more preferably at least about 95% of one enantiomer, or most preferably at least about 99% of one enantiomer. Similarly, the phrase "substantially free of other optical isomers" means that the composition comprises at most about 15% of another enantiomer or diastereomer, more preferably at most about 10% of the other enantiomer or diastereomer, even more preferably at most about 5% of the other enantiomer or diastereomer, and most preferably at most about 1% of the other enantiomer or diastereomer.

"preventing" or "preventing" includes: (1) inhibiting the onset of disease in a subject or patient at risk for and/or susceptible to disease but not yet experiencing or exhibiting any or all of the pathology or symptomology of disease, and/or (2) slowing the onset of disease pathology or symptomology in a subject or patient at risk for and/or susceptible to disease but not yet experiencing or exhibiting any or all of the pathology or symptomology of disease.

By "prodrug" is meant a compound that is metabolically converted in vivo to the disclosed inhibitors. The prodrug itself may or may not also have activity against a particular target protein. For example, a compound containing a hydroxyl group may be administered as an ester, which is converted to a hydroxyl compound in vivo by hydrolysis. Suitable esters that can be converted in vivo to hydroxy compounds include acetate, citrate, lactate, phosphate, tartrate, malonate, oxalate, salicylate, propionate, succinate, fumarate, maleate, methylene-bis- β -hydroxynaphthoate, cholate, isethionate, di-p-methylbenzoyl tartrate, mesylate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclohexylsulfamate, quinic acid ester, amino acid ester, and the like. Similarly, compounds containing amine groups may be administered as amides that are converted to amine compounds by hydrolysis in vivo.

The term "saturated" when referring to an atom means that the atom is attached to other atoms only by single bonds.

"stereoisomers" or "optical isomers" are isomers of a given compound in which the same atom is bonded to the same other atom, but those atoms are not in the same configuration in three-dimensional space. "enantiomers" are stereoisomers of a given compound that are mirror images of each other, as are left and right handed. "diastereomer" is a stereoisomer of a given compound that is not an enantiomer.

The present invention contemplates that, for any chiral stereocenter or axis for which stereochemistry is not yet defined, the chiral stereocenter or axis may exist in its R form, S form, or as a mixture of R and S forms, including racemic and non-racemic mixtures.

By "substituents converted to hydrogen in vivo" is meant by enzymology orChemical means, including but not limited to hydrolysis and hydrogenolysis into hydrogen atoms of any group. Examples include acyl groups, groups having oxycarbonyl groups, amino acid residues, peptide residues, o-nitrophenylsulfinyl (sulfenyl), trimethylsilyl, tetrahydropyranyl, diphenylphosphinyl (phosphinyl), hydroxy or alkoxy substituents on imino groups, and the like. Examples of acyl groups include formyl, acetyl, trifluoroacetyl, and the like. Examples of the group having an oxycarbonyl group include ethoxycarbonyl group, t-butoxycarbonyl (mono C (O) OC (CH) ₃)₃) Benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, vinyloxycarbonyl, β - (p-methylbenzenesulfonyl) ethoxycarbonyl, and the like. Suitable amino acid residues include, but are not limited to, Gly (glycine), Ala (alanine), Arg (arginine), Asn (asparagine), Asp (aspartic acid), Cys (cysteine), Glu (glutamic acid), His (histidine), Ile (isoleucine), Leu (leucine), Lys (lysine), Met (methionine), Phe (phenylalanine), Pro (proline), Ser (serine), Thr (threonine), Trp (tryptophan), Tyr (tyrosine), Val (valine), Nva (norvaline), Hse (homoserine), 4-Hyp (4-hydroxyproline), 5-Hyl (5-hydroxylysine), Orn (ornithine), and β -Ala residues. Examples of suitable amino acid residues also include amino acid residues protected with a protecting group. Examples of suitable protecting groups include those typically employed in peptide synthesis, including acyl groups (e.g., formyl and acetyl), arylmethyloxycarbonyl groups (e.g., benzyloxycarbonyl and p-nitrobenzyloxycarbonyl), t-butyloxycarbonyl (-C (O) OC (CH)₃)₃) And so on. Suitable peptide residues include peptide residues comprising 2 to 5 and optionally amino acid residues. The residues of these amino acids or peptides may exist in stereochemical configuration in D-form, L-form or mixtures thereof. Furthermore, the amino acid or peptide residue may have asymmetric carbon atoms. Examples of suitable amino acid residues having asymmetric carbon atoms include Ala, Leu, Phe, Trp, Nva, Val, Met, Ser, Lys, Thr and Tyr residues. Examples of peptide residues having asymmetric carbon atoms include peptide residues having one or more than one constituent amino acid residue containing an asymmetric carbon atom. Examples of suitable amino acid protecting groups include those in peptidomimetics Typically employed ones include acyl groups (e.g., formyl and acetyl), arylmethyloxycarbonyl groups (e.g., benzyloxycarbonyl and p-nitrobenzyloxycarbonyl), t-butyloxycarbonyl group (-C (O)) OC (CH)₃)₃) And so on. Other examples of substituents "converted to hydrogen in vivo" include reductively eliminable hydrogenolyzable groups. Suitable examples of reductively eliminable hydrogenolyzable groups include, but are not limited to, arylsulfonyl (e.g., o-toluenesulfonyl); methyl substituted with phenyl or benzyloxy (e.g., benzyl, trityl, and benzyloxymethyl); arylmethoxycarbonyl (e.g., benzyloxycarbonyl and o-methoxybenzyloxycarbonyl); and haloethoxycarbonyl groups (e.g., β, β, β -trichloroethoxycarbonyl and β -iodoethoxycarbonyl).

By "therapeutically effective amount" or "pharmaceutically effective amount" is meant an amount sufficient to effect such treatment of a disease when administered to a subject or patient for the purpose of treating the disease.

"treating" or "treatment" includes (1) inhibiting a disease in a subject or patient experiencing or exhibiting a disease pathology or syndrome (e.g., arresting further development of the pathology and/or syndrome), (2) ameliorating a disease in a subject or patient experiencing or exhibiting a disease pathology or syndrome (e.g., reversing the pathology and/or syndrome), and/or (3) causing measurable regression of a disease in a subject or patient experiencing or exhibiting a disease pathology or syndrome.

As used herein, the term "water soluble" means that the compound is dissolved in water to the extent of at least 0.010 moles/liter or classified as soluble according to literature knowledge.

Other abbreviations used herein are as follows: DMSO, dimethyl sulfoxide; NO, nitric oxide; iNOS, inducible nitric oxide synthase; COX-2, cyclooxygenase-2; NGF, nerve growth factor; IBMX, isobutylmethylxanthine; FBS, peptide bovine serum; GPDH, glycerol 3-phosphate dehydrogenase; RXR, retinoid X receptor; TGF- β, transforming growth factor- β; IFN gamma or IFN-gamma, interferon-gamma; LPS, bacterial endotoxin lipopolysaccharide; TNF α or TNF- α, tumor necrosis factor- α; IL-1 β, interleukin-1 β; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; MTT, 3- [4, 5-dimethylthiazol-2-yl ] -2, 5-diphenyltetrazolium bromide; TCA, trichloroacetic acid; HO-1, inducible heme oxygenase.

The above definitions supersede any conflicting definition in any reference incorporated herein by reference. However, the fact that certain terms are defined should not be understood to imply that any terms not defined are ambiguous. Rather, it is intended that all terms used be interpreted as describing the invention under such conditions that the skilled artisan would understand the scope and practice of the disclosure.

Synthesis Process

One of the anti-inflammatory pharmacophores of the present invention can be represented by the following sections:

x, R therein₁And R₂As defined above and in the claims below. This pharmacophore is present in a variety of synthetic triterpenoids, such as Honda et al, (2000 a); honda et al, (2000 b); honda et al, (2002); and those described in U.S. application No. 11/941,820, each of which is incorporated herein by reference. Other compounds comprising such pharmacophores are described in the following applications filed concurrently with the present application, each of which is incorporated herein by reference: U.S. patent application filed by Eric Anderson, Xin Jiang, Xiaofeng Liu, mean Visnick, entitled "antipixidant informationmodulators: oleanolic Acid Derivatives With preservation in the C-Ring (antioxidant inflammation regulator: Oleanolic Acid derivative With saturated C-Ring)', filed on 4/20 days in 2009; U.S. patent applications filed by Eric Anderson, Xin Jiang and mean Visnick, entitled "angular information Modulators: oleanolic Acid Derivatives with Amini and other Modifications At C-17 (antioxidant inflammation modulators: oleanolic acid derivatives with amino groups and other Modifications At C-17) ", filed on 4/20 days 2009; U.S. patent application filed by Xin Jiang, Jack Greiner, Lester L.Maravetz, Stephen S.Szucs, Melean Visnick, entitled "adjustment information modules: novel Derivatives of Oleanolic Acid (antioxidant inflammation regulator: Novel Oleanolic Acid derivative)', filed on 4/20/2009; and U.S. patent applications filed by Xin Jiang, Xioaveng Liu, Jack Greiner, Stephen S.Szucs, Melean Visnick, entitled "Artificial information modules: c-17Homologated Oleanolic Acid Derivatives (antioxidant inflammation modulators: C-17Homologated Oleanolic Acid Derivatives) "filed on 4/20 days 2009. Many of these compounds ("mentioned compounds") have been shown to have diverse anti-inflammatory related activities, including, for example, antiproliferative and/or antioxidant activities such as the in vitro and in vivo induction of heme oxygenase-1 (HO-1), induction of CD11B expression, inhibition of iNOS induction, inhibition of COX-2 induction, inhibition of NO production, induction of cancer cell apoptosis, inhibition of NF-. kappa.B, activation of JNK pathway, and phase 2 induction (NAD (P) H-quinone oxidoreductase and HO-1 elevation). Induction of the Phase 2 response is associated with activation of the transcription factor Nrf2, it has been demonstrated that transcription factor Nrf2 activates the Antioxidant Response Element (ARE) of the promoter region of many antioxidant, anti-inflammatory and cytoprotective genes, and that Phase 2 activation is highly correlated with effective inhibition of NO production in activated macrophages (e.g., Dinkova-kostowa et al, proc.natl.acad.sci.u S a.2005; 102 (12): 4584-9). Since (i) the compounds of the present invention share a pharmacophore with many of the mentioned compounds, and (ii) like the mentioned compounds, the compounds of the present invention also demonstrate inhibition of NO production, it is also possible that the compounds of the present invention exhibit one or more of the anti-inflammatory activities exhibited by the mentioned compounds. As shown herein, the introduction of this pharmacophore into molecules that exhibit low or modest potency as inhibitors of NO production consistently provides compounds with significantly increased potency. For example, in the NO assay, Compound C 0009 with an IC of 100nM₅₀And curcumin IC as a parent compound₅₀At 1.5. mu.M. Similarly, DHEA is inactive in the NO assay (IC)₅₀> 10. mu.M), IC for Compound D0018₅₀Is 130 nM. Preferably, the improved compounds will have an IC of less than 1.25. mu.M₅₀. Still more preferably, the improved compounds will have an IC of less than 500nM₅₀. In certain embodiments, suitable modifying compounds contemplated herein include, but are not limited to, triterpenoids (non-limiting examples of which include glycyrrhetinic acid, boswellic acid, colatodiol, calendula glycol, and tetronic acid (moronic acid)), saponins (e.g., ginsenosides), avicin, resveratrol, curcumin, gossypol, epigallocatechin-3-gallate (EGCG), gossypol, lapachol, other flavonoids (non-limiting examples of which include quercetin, daidzein, luteolin, coumarin, wogonin, and baicalin), Dehydroandrosterone (DHEA), cholic acid, deoxycholic acid, ginsenosides (e.g., 20(S) -ginsenoside), silybin, anthocyanins, avenanthramide, cucurbitin, aloesin, aloemodin (DHEA), and mixtures thereof, And/or tubeimoside A.

Non-limiting examples of compounds of the present disclosure that include this particular pharmacophore include:

the compounds of the present disclosure were made using the methods outlined below and in the examples section (examples 2 and 3). These methods can be modified and optimized using organic chemistry principles and techniques as applied by those skilled in the art. Such principles and techniques are described, for example, in March's Advanced Organic Chemistry: reactions, Mechanisms, and Structure (2007), which is incorporated herein by reference.

Biological Activity

The compounds of the disclosure have been tested for inhibition of NO production. The results of these experiments are shown in the figure and in table 1 below. Experimental details are provided in example 1.

TABLE 1 suppression of IFN γ -induced NO production

Diseases associated with inflammation and/or oxidative stress

Inflammation is a biological process that provides protection against infectious or parasitic organisms and repairs damaged tissues. Inflammation is generally characterized by local vasodilation, redness, swelling, and pain, recruitment of leukocytes to the site of infection or injury, production of inflammatory cytokines such as TNF- α and IL-1, and production of reactive oxygen or nitrogen clusters, such as hydrogen peroxide, superoxide, and peroxynitrite. In the later stages of inflammation, tissue remodeling, angiogenesis and scarring (fibrosis) may occur as part of the wound healing process. Under normal circumstances, the inflammatory response is regulatory and transient, and subsides in a coordinated fashion once the infection or injury has been adequately treated. However, if the regulatory mechanisms fail, acute inflammation can become severe and life threatening. Alternatively, inflammation can become chronic and lead to cumulative tissue damage or systemic complications.

Many serious and intractable human diseases involve dysregulation of inflammatory processes, including diseases such as cancer, atherosclerosis and diabetes, which are not conventionally considered inflammatory diseases. In the case of cancer, the inflammatory process is associated with tumor formation, progression, metastasis and resistance to treatment. Atherosclerosis, which has long been regarded as a disorder of lipid metabolism, is now understood to be primarily an inflammatory disease in which activated macrophages play an important role in the formation and eventual rupture of atherosclerotic plaques. Activation of inflammatory signaling pathways has also been shown to play a role in the development of insulin resistance and in peripheral tissue damage associated with diabetic hyperglycemia. Overproduction of reactive oxygen and nitrogen clusters such as superoxide, hydrogen peroxide, nitric oxide and peroxynitrite is a hallmark of inflammatory diseases. Evidence of deregulated peroxynitrite production has been reported in a wide variety of diseases (Szabo et al, 2007; Schulz et al, 2008; Forstermann, 2006; Pall, 2007).

Autoimmune diseases such as rheumatoid arthritis, lupus, psoriasis and multiple sclerosis involve inappropriate and chronic activation of inflammatory processes in affected tissues, resulting from dysfunction of self-to non-self recognition and response mechanisms in the immune system. In neurodegenerative diseases such as alzheimer's disease and parkinson's disease, nerve damage is associated with activation of microglia and increased levels of proinflammatory proteins such as Inducible Nitric Oxide Synthase (iNOS). Chronic organ failure, such as renal failure, heart failure and chronic obstructive pulmonary disease, is closely associated with the presence of chronic oxidative stress and inflammation, which leads to the development of fibrosis and ultimately the loss of organ function.

Many other conditions involve oxidative stress and inflammation in affected tissues, including inflammatory bowel disease; inflammatory skin diseases; mucositis associated with radiation therapy and chemotherapy; eye diseases such as uveitis, glaucoma, macular degeneration, and various forms of retinopathy; graft failure and rejection; ischemia-reperfusion injury; chronic pain; degenerative diseases of bones and joints, including osteoarthritis and osteoporosis; asthma and cystic fibrosis; epilepsy; and neuropsychiatric disorders including schizophrenia, depression, bipolar disorder, post-traumatic stress disorder, attention deficit disorder, autism spectrum disorder, and eating disorders such as anorexia nervosa. Dysregulation of inflammatory signaling pathways is believed to be a major factor in muscle wasting diseases, including muscular dystrophy and the pathology of various forms of cachexia.

A variety of life-threatening acute conditions are also involved in dysregulated inflammatory signaling, including acute organ failure involving the pancreas, kidneys, liver or lungs, myocardial infarction or acute coronary syndrome, stroke, septic shock, trauma, severe burns and anaphylaxis.

Many complications of infectious diseases also involve dysregulation of the inflammatory response. While the inflammatory response can kill invading pathogens, a severe inflammatory response can also be completely destructive and in some cases can be a major source of damage in infected tissue. In addition, severe inflammatory responses can lead to systemic complications due to the overproduction of inflammatory cytokines such as TNF- α and IL-1. It is believed that this is a cause of death due to severe influenza, severe acute respiratory syndrome, and sepsis.

Aberrant or overexpression of either iNOS or the cyclooxygenase-2 (COX-2) enzyme has been implicated in the pathogenesis of a number of diseases. For example, it has been shown that NO is a potent mutagen (Tamir and Tannebaum, 1996), and that nitric oxide can also activate COX-2(Salvemini et al, 1994). Furthermore, iNOS is significantly increased in large intestine colon tumors induced by the carcinogen azoxymethane (Takahashi et al, 1997). A series of synthetic oleanolic acid triterpenoid analogs have been shown to be potent inhibitors of cellular inflammatory processes, such as the induction of Inducible Nitric Oxide Synthase (iNOS) and COX-2 by IFN-. gamma.in mouse macrophages. See Honda et al (2000 a); honda et al (2000b), and Honda et al (2002), all of which are incorporated herein by reference.

In one aspect, the compounds of the invention are characterized by their inhibitory ability to the induction of nitric oxide production in macrophage-derived RAW 264.7 cells by gamma-interferon. They are further characterized by their ability to induce the expression of antioxidant proteins such as NQO1 and to reduce the expression of pro-inflammatory proteins such as COX-2 and Inducible Nitric Oxide Synthase (iNOS). These properties are associated with the treatment of a wide range of diseases involving disturbances of oxidative stress and inflammatory processes, including cancer, mucositis from radiotherapy or chemotherapy, autoimmune diseases, cardiovascular diseases including atherosclerosis, ischemia-reperfusion injury, acute and chronic organ failure including renal failure and heart failure, respiratory diseases, diabetes and diabetic complications, severe allergies, transplant rejection, graft-versus-host disease, neurodegenerative diseases, eye and retinal diseases, acute and chronic pain, degenerative bone diseases including osteoarthritis and osteoporosis, inflammatory bowel disease, dermatitis and other skin diseases, sepsis, burns, epilepsy and neuropsychiatric disorders.

Without wishing to be bound by theory, activation of the antioxidant/anti-inflammatory Keap1/Nrf2/ARE pathway is believed to be related to both the anti-inflammatory and anti-carcinogenic properties of the oleanolic acid derivatives of the present invention.

In another aspect, the compounds of the invention may be used to treat a subject suffering from a disease caused by an elevated level of oxidative stress in one or more tissues. Oxidative stress results from abnormally high or sustained long levels of reactive oxygen species such as superoxide, hydrogen peroxide, nitric oxide and peroxynitrite (formed by the nitric oxide and superoxide reactions). Oxidative stress may be accompanied by acute or chronic inflammation. Oxidative stress may be caused by mitochondrial dysfunction, activation of immune cells such as macrophages and neutrophils, acute exposure to external agents such as ionizing radiation or cytotoxic chemotherapeutic agents (e.g., doxorubicin), trauma or other acute tissue injury, ischemia/reperfusion, poor circulation or anemia, local or systemic hypoxia or hyperoxia, elevated levels of inflammatory cytokines and other inflammation-related proteins, and/or other abnormal physiological states such as hyperglycemia or hypoglycemia.

Stimulation of Nrf2 pathway target genes has been shown to induce the expression of heme oxygenase (HO-1) with significant therapeutic effects in a number of animal models of such diseases, including models of myocardial infarction, renal failure, graft failure and rejection, stroke, cardiovascular disease and autoimmune disease (e.g., Sacerdoti et al, 2005; Abraham and Kappas, 2005; Bach, 2006; arajo et al, 2003; Liu et al, 2006; Ishikawa et al, 2001; Kruger et al, 2006; Satoh et al, 2006; Zhou et al, 2005; Morse and Choi, 2002). This enzyme breaks down free heme into iron, carbon monoxide (CO) and biliverdin, which is subsequently converted into the potent antioxidant molecule bilirubin.

In another aspect, the compounds of the present invention may be used for the prevention or treatment of acute and chronic tissue injury or organ failure caused by oxidative stress exacerbated by inflammation. Examples of diseases in this class include: heart failure, liver failure, graft failure and rejection, renal failure, pancreatitis, fibrotic lung diseases (especially cystic fibrosis and COPD), diabetes (including complications), atherosclerosis, ischemia-reperfusion injury, glaucoma, stroke, autoimmune diseases, autism, macular degeneration, and muscular dystrophy. For example, in the case of autism, studies have shown that increased oxidative stress in the central nervous system contributes to disease progression (Chauhan and Chauhan, 2006).

Evidence also links oxidative stress and inflammation to the development and pathology of many other disorders of the central nervous system, including neuropathies such as psychosis, major depression, and bipolar disorder; epilepsy such as epilepsy; pain and sensory syndromes such as migraine, neuropathic pain or tinnitus; and behavioral syndromes such as attention deficit disorder. See, e.g., Dickerson et al, 2007; hanson et al, 2005; kendall-tatkett, 2007; lencz et al, 2007; dudhgaonkar et al, 2006; lee et al, 2007; morris et al, 2002; ruster et al, 2005; McIver et al, 2005; sarchielli et al, 2006; kawakami et al, 2006; ross et al, 2003, which are incorporated herein by reference in their entirety. For example, elevated levels of inflammatory cytokines, including TNF, interferon-gamma and IL-6, are associated with most psychological disorders (Dickerson et al, 2007). Microglial activation is also associated with most psychological disorders. Therefore, down-regulation of inflammatory cytokines and inhibition of excessive activation of microglia would be beneficial to patients with schizophrenia, major depression, bipolar disorder, autism spectrum disorder, and other neuropsychiatric conditions.

Thus, in pathologies involving only oxidative stress or oxidative stress exacerbated by inflammation, treatment may comprise administering to the subject a therapeutically effective amount of a compound of the invention, such as those described above or throughout this specification. The treatment may be administered prophylactically (e.g., organ transplantation or radiation therapy to a cancer patient) prior to a predictable oxidative stress state, or it may be administered therapeutically in situations involving established oxidative stress and inflammation.

The compounds of the present invention may be used in general for the treatment of inflammatory diseases such as sepsis, dermatitis, autoimmune diseases and osteoarthritis. In one aspect, the compounds of the invention may be used to treat inflammatory and/or neuropathic pain, for example, by inducing Nrf2 and/or inhibiting NF- κ B.

In one aspect, the compounds of the present invention may function as Antioxidant Inflammation Modulators (AIMs) with potent anti-inflammatory properties that mimic the biological activity of cyclopentenone prostaglandins (cypgs). In one embodiment, the compounds of the invention may be used to control the production of pro-inflammatory cytokines by selectively targeting Regulatory Cysteine Residues (RCRs) on proteins that modulate the transcriptional activity of redox-sensitive transcription factors. It has been demonstrated that activation of RCR by cyPG or AIM initiates a pro-resolution procedure in which the activities of antioxidant and cytoprotective transcription factor Nrf2 are effectively induced and the activities of pro-oxidant and pro-inflammatory transcription factors NF-. kappa.B and STAT are suppressed. This increases the production of antioxidant and reducing molecules (e.g., NQO1, HO-1, SOD1, and/or γ -GCS) and/or decreases the production of oxidative stress and pro-oxidant and pro-inflammatory molecules (e.g., iNOS, COX-2, and/or TNF- α).

In some embodiments, the compounds of the invention may be used for the treatment and prevention of diseases, such as cancer, inflammation, alzheimer's disease, parkinson's disease, multiple sclerosis, autism, amyotrophic lateral sclerosis, autoimmune diseases such as rheumatoid arthritis, lupus and MS, inflammatory bowel disease, all other diseases whose pathogenesis is believed to involve overproduction of either nitric oxide or prostaglandins, and pathologies involving oxidative stress alone or exacerbated by inflammation.

Another aspect of inflammation is the production of inflammatory prostaglandins such as prostaglandin E. These molecules promote vasodilation, plasma extravasation, local pain, elevated temperature and other symptoms of inflammation. The enzyme COX-2 in its inducible form is associated with their production and high levels of COX-2 are found in inflamed tissues. Thus, inhibition of COX-2 can alleviate many symptoms of inflammation and many important anti-inflammatory drugs (e.g., ibuprofen and celecoxib) act by inhibiting COX-2 activity. However, recent studies have shown that a class of cyclopentenone prostaglandins (cypgs) (e.g., 15-deoxy prostaglandin J2, also known as PGJ2) play a role in stimulating the resolution of a coordinated fashion of inflammation (e.g., Rajakariar et al, 2007). COX-2 is also associated with the production of cyclopentenone prostaglandins. Thus, inhibition of COX-2 would interfere with complete resolution of inflammation, potentially promoting sustained retention of activated immune cells in the tissue and leading to chronic "stasis" inflammation. This result has led to an increased incidence of cardiovascular disease in patients who have been administered selective COX-2 inhibitors over a prolonged period of time.

In one aspect, the compounds of the invention are useful for controlling the production of proinflammatory cytokines in a cell by selectively activating Regulatory Cysteine Residues (RCRs) on proteins that modulate the activity of redox-sensitive transcription factors. It has been shown that activation of RCR by cyPG initiates a pro-resolution procedure in which the activities of antioxidant and cytoprotective transcription factor Nrf2 are effectively induced and the activities of pro-oxidant and pro-inflammatory transcription factors NF-. kappa.B and STAT are suppressed. In some embodiments, this increases the production of antioxidants and reducing molecules (NQO1, HO-1, SOD1, γ -GCS) and decreases the production of oxidative stress and pro-oxidant and pro-inflammatory molecules (iNOS, COX-2, TNF- α). In some embodiments, the compounds of the invention will revert to a non-inflammatory state by promoting resolution of inflammation and limiting host excessive tissue damage to cells with inflammatory events.

A. Cancer treatment

In addition, the compounds of the present disclosure may be used to induce tumor cell apoptosis, induce cell differentiation, inhibit cancer cell proliferation, inhibit inflammatory responses, and/or play a role in chemopreventive capacity. For example, the present invention provides novel compounds having one or more of the following properties: (1) the ability to induce apoptosis and differentiation in malignant and non-malignant cells, (2) activity as inhibitors of proliferation in many malignant or pre-malignant cells at submicromolar or nanomolar levels, (3) the ability to suppress de novo synthesis of inflammatory enzymes Inducible Nitric Oxide Synthase (iNOS), (4) the ability to inhibit activation of NF- κ B, and (5) the ability to induce expression of heme oxygenase-1 (HO-1).

Levels of iNOS and COX-2 are elevated in certain cancers and have been implicated in carcinogenesis, and COX-2 inhibitors have been shown to reduce the incidence of primary colon adenomas in humans (Rostom et al, 2007; Brown and DuBois, 2005; Crowlel et al, 2003). iNOS is expressed in Myeloid Derived Suppressor Cells (MDSC) (Angulo et al, 2000), and COX-2 activity in cancer cells has been shown to result in prostaglandin E₂(PGE₂) Has demonstrated production of prostaglandin E₂Arginase expression was induced in MDSCs (Sinha et al, 2007). Arginase and iNOS are enzymes that utilize L-arginine as a substrate and produce L-ornithine and urea and L-citrulline and NO, respectively. Depletion of arginine by MDSCs in the tumor microenvironment, in combination with the production of NO and peroxynitrite, has been shown to inhibit proliferation and induce apoptosis in T cells (Bronte et al, 2003). Inhibition of COX-2 and iNOS has been shown to reduce MDSC accumulation, restore cytotoxic activity of tumor-associated T cells, and delay tumor growth (Sinha et al, 2007; Mazzoni et al, 2002; Zhou et al, 2007).

Inhibition of NF-. kappa.B and JAK/STAT signaling pathways has been implicated as a strategy for inhibiting proliferation and inducing apoptosis in cancer epithelial cells. Activation of STAT3 and NF-. kappa.B has been shown to lead to suppression of apoptosis in cancer cells, and promotion of proliferation, invasion, and metastasis. Many target genes involved in these processes have been shown to be transcriptionally regulated by NF-. kappa.B and STAT3 (Yu et al, 2007).

In addition to their direct role in cancer epithelial cells, NF- κ B and STAT3 also play important roles in other cells present in the tumor microenvironment. Experiments in animal models have demonstrated that NF-. kappa.B is required in both cancer and hematopoietic cells to transmit the effects of inflammation on cancer initiation and progression (Greten et al, 2004). Inhibition of NF-. kappa.B in cancer and myeloid cells reduces the number and size of tumors formed, respectively. Activation of STAT3 in cancer cells results in the production of several cytokines (IL-6, IL-10) that suppress the maturation of tumor-associated Dendritic Cells (DCs). In addition, STAT3 is activated by these cytokines within the dendritic cells themselves. STAT3 inhibition restores DC maturation, promotes anti-tumor immunity, and inhibits tumor growth in a mouse cancer model (Kortylewski et al, 2005).

B. Treatment of multiple sclerosis

The compounds and methods of the invention may be used to treat Multiple Sclerosis (MS) patients. MS is known to be an inflammatory disorder of the central nervous system (Williams et al, 1994; Merrill and Benvenist, 1996; Genain and Nauser, 1997). Based on several studies, there is evidence to suggest that inflammatory, oxidative and/or immunological mechanisms are involved in the pathogenesis of Alzheimer's Disease (AD), Parkinson's Disease (PD), Amyotrophic Lateral Sclerosis (ALS) and MS (Bagasra et al, 1995; McGeer and McGeer, 1995; Simonian and Coyle, 1996; Kaltschmidt et al, 1997). Both activated astrocytes and activated microglia are involved in causing neurodegenerative diseases (NDD) and neuroinflammatory diseases (NID); here, a particularly potent microglia is one that synthesizes both NO and prostaglandin, which are the products of the respective iNOS and COX-2 enzymes. De novo formation of these enzymes may be driven by inflammatory cytokines such as interferon-gamma or interleukin-1. In turn, overproduction of NO can lead to inflammatory cascades and/or oxidative damage in cells and tissues of many organs, including neurons and oligodendrocytes of the nervous system, with consequent manifestations of AD and MS, and possibly PD and ALS (Coyle and Puttfarcken, 1993; Beal, 1996; Merrill and Benvenist, 1996; Simonia and Coyle, 1996; Vodovotz et al, 1996). Epidemiological data suggest that chronic use of NSAIDs that block the synthesis of prostaglandins from arachidonic acid significantly reduces the risk of developing AD (McGeer et al, 1996; Stewart et al, 1997). Thus, agents that block the formation of NO and prostaglandins may be used in methods of preventing and treating NDD. The ability to cross the blood-brain barrier is typically required for successful therapeutic candidates for the treatment of this disease. See, e.g., U.S. patent publication No. 2009/0060873, which is incorporated by reference herein in its entirety.

C. Inflammation of nerve

Neuroinflammation is a generalization of the concept that the responses and actions of microglia and astrocytes in the central nervous system have essentially inflammation-like characteristics, and that these responses are important for the pathogenesis and progression of a wide variety of neurological disorders. This concept originates from the field of Alzheimer's disease (Griffin et al, 1989; Rogers et al, 1988), where it revolutionized our understanding of the disease (Akiyama et al, 2000). These concepts have been extended to other neurodegenerative diseases (Eikelenboom et al, 2002; Ishizawa and Dickson, 2001), ischemic/toxic diseases (Gehrmann et al, 1995; Touzani et al, 1999), tumor biology (Graeber et al, 2002) and even normal brain development.

Neuroinflammation incorporates a broad spectrum of complex cellular responses including activation of microglia and astrocytes and induction of cytokines, chemokines, complement proteins, acute phase proteins, oxidative damage and related molecular processes. These events will have a deleterious effect on neuronal function, leading to neuronal damage, further to glial activation, and ultimately to neurodegeneration.

Based on the experimental results obtained, including those given in this application, the compounds and methods of the present invention may be used to treat patients with neuroinflammation.

D. Treatment of renal failure

Another aspect of the present disclosure relates to novel methods and compounds for the treatment and prevention of renal disease. See U.S. patent application 12/352,473, which is incorporated herein by reference in its entirety. Renal failure, which results from inadequate clearance of metabolic wastes in the blood and abnormal electrolyte concentrations in the blood, is an important medical problem worldwide, especially in developed countries. Diabetes and hypertension are among the most important causes of chronic renal failure (CKD), but are also associated with other diseases such as lupus. Acute renal failure can result from contact with certain drugs (e.g., acetaminophen) or toxic chemicals, or from ischemia-reperfusion injury associated with shock or surgery, such as transplantation, and can lead to chronic renal failure. In many patients, renal failure can progress to a stage where the patient requires routine dialysis or kidney transplantation to sustain life. These processes are both highly invasive and associated with significant side effects and quality of life issues. Although effective treatments exist for some of the complications of renal failure, such as hyperparathyroidism and hyperphosphatemia, there is no available method to stop or reverse the underlying progression of renal failure. Thus, agents that can improve compromised kidney function would represent a significant advance in the treatment of renal failure.

Inflammation contributes significantly to the pathology of CKD. There is also a strong mechanistic link between oxidative stress and renal dysfunction. The NF-. kappa.B signaling pathway plays an important role in the progression of CKD, because NF-. kappa.B regulates the transcription of MCP-1, a chemokine responsible for recruiting monocytes/macrophages to cause an inflammatory response and ultimately damage the kidney (Wardle, 2001). The Keap1/Nrf2/ARE pathway controls transcription of several genes encoding antioxidant enzymes, including heme oxygenase-1 (HO-1). Excision of the Nrf2 gene in female mice resulted in the development of lupus-like glomerulonephritis (Yoh et al, 2001). Furthermore, several studies have demonstrated that HO-1 expression is induced in response to renal injury and inflammation and that the enzyme and its products bilirubin and carbon monoxide play a protective role in the kidney (Nath et al, 2006).

The glomerulus and the surrounding Bowman's capsule constitute the basic functional unit of the kidney. Glomerular Filtration Rate (GFR) is a standard measure of kidney function. Creatinine clearance is commonly used to measure GFR. However, serum creatinine levels are often used as a surrogate measure of creatinine clearance. For example, excessive levels of serum creatinine are generally recognized as indicative of insufficient kidney function, and a decrease in serum creatinine over time is recognized as indicative of improved kidney function. Normal levels of creatinine in the blood are about 0.6 to 1.2 milligrams (mg) per deciliter (dl) in adult males and 0.5 to 1.1 milligrams per deciliter in adult females.

Acute Kidney Injury (AKI) can occur following ischemia-reperfusion, following treatment with certain pharmaceutical agents such as cisplatin and rapamycin, and following intravenous injection of radiocontrast agents for medical imaging. As in CKD, inflammation and oxidative stress contribute to the pathology of AKI. The molecular mechanism of radiocontrast induced nephropathy (RCN) is not well understood; however, it is likely that the combination of events including prolonged vasoconstriction, impaired renal self-regulation and direct contrast agent toxicity all contribute to renal failure (Tumlin et al, 2006). Vasoconstriction results in reduced renal blood flow and produces ischemia-reperfusion and the production of reactive oxygen species. HO-1 is strongly induced in these conditions and strong induction of HO-1 has been demonstrated to prevent ischemia-reperfusion injury in several different organs, including the kidney (Nath et al, 2006). In particular, the induction of HO-1 has been shown to be protective in a rat model of RCN (Goodman et al, 2007). Reperfusion also causes an inflammatory response, in part through activation of NF- κ B signaling (Nichols, 2004). Targeting NF- κ B has been proposed as a therapeutic strategy to prevent organ damage (Zingarelli et al, 2003).

Based on the experimental results obtained, including those given in this application, the compounds and methods of the present invention can be used to treat patients with renal failure.

E. Cardiovascular diseases

The compounds and methods of the invention may be used to treat patients suffering from cardiovascular disease. See U.S. patent application 12/352,473, which is incorporated herein by reference in its entirety. Cardiovascular (CV) disease is the leading cause of death worldwide, and in many developed countries the leading cause of death. The etiology of CV disease is complex, but most causes are associated with inadequate or complete disruption of blood supply to critical organs or tissues. Such diseases often result from the rupture of one or more atherosclerotic plaques, which leads to the formation of a thrombus that blocks blood flow in critical blood vessels. Such thrombosis is a major cause of heart attack in which one or more coronary arteries are occluded and blood flow to the heart itself is interrupted. The resulting ischemia highly damages cardiac tissue due to both hypoxia during the ischemic event and excessive formation of free radicals from after restoration of blood flow (a phenomenon known as ischemia-reperfusion injury). Similar damage occurs in the brain during thrombotic stroke when cerebral arteries or other major blood vessels become occluded by thrombosis. In contrast, hemorrhagic stroke involves the rupture of blood vessels and the influx of blood into the surrounding brain tissue. This can result in oxidative stress in the immediate area of bleeding due to the presence of large amounts of free heme and other active species and other ischemia of other parts of the brain due to impaired blood flow. Subarachnoid hemorrhage, often accompanied by cerebral vasospasm, also leads to ischemia/reperfusion injury in the brain.

Alternatively, atherosclerosis is so extensive in critical blood vessels that it develops into stenosis (narrowing of the artery) and blood flow to critical organs, including the heart, is chronically inadequate. Such chronic ischemia can result in many types of end organ damage, including cardiac hypertrophy associated with congestive heart failure.

When physical defects or damage to the inner layer of the artery (endothelium) trigger an inflammatory response, including proliferation of vascular smooth muscle and infiltration of leukocytes into the affected area, a potential defect occurs leading to atherosclerosis in various forms of cardiovascular disease. Eventually, a complicated lesion called atherosclerotic plaque may form, consisting of the above cells in combination with deposits of cholesterol-carrying lipoproteins and other substances (e.g., Hansson et al, 2006).

Drug treatment of cardiovascular disease includes prophylactic treatment, such as the use of drugs aimed at lowering blood pressure or circulating levels of cholesterol and lipoproteins, and treatment designed to reduce platelet and other blood cell attachment tendencies (and thus reduce platelet aggregation rates and thrombosis risk). More recently, drugs such as streptokinase and tissue plasminogen activator have been introduced and used to dissolve thrombi and restore blood flow. Surgical treatments include coronary artery bypass grafting to create an alternative blood supply, balloon angioplasty to compress plaque tissue and increase the diameter of the arterial lumen, and carotid endarterectomy to remove plaque tissue in the carotid artery. Such treatments, particularly balloon angioplasty, are accompanied by the use of stents, expandable mesh tubes designed to support the arterial wall of the affected area and maintain vessel patency. Recently, the use of drug eluting stents has become common to prevent post-operative restenosis (restenosis of the artery) in the affected area. These devices are metal stents overcoated with a biocompatible polymeric matrix containing a drug that inhibits cell proliferation (e.g., paclitaxel or rapamycin). The polymer allows for slow, localized release of the drug in the affected area, minimizing non-target tissue contact. Despite the obvious benefits of such treatments, mortality from cardiovascular disease remains high and clearly does not meet the needs of cardiovascular disease treatment.

As mentioned above, it has been demonstrated that the induction of HO-1 is beneficial in many cardiovascular disease models and that low levels of HO-1 expression are clinically relevant for high risk of CV disease. Thus, the compounds of the present invention may be used to treat or prevent a wide variety of cardiovascular diseases, including but not limited to atherosclerosis, hypertension, myocardial infarction, chronic heart failure, stroke, subarachnoid hemorrhage and restenosis.

F. Diabetes mellitus

Diabetes mellitus is a complex disease characterized by the inability of the body to regulate circulating levels of glucose. See U.S. patent application 12/352,473, which is incorporated herein by reference in its entirety. This deficiency may result from insulin deficiency, a peptide hormone that regulates glucose production and absorption in a variety of tissues. Insulin deficiency impairs the ability of muscle, fat and other tissues to properly absorb glucose, resulting in hyperglycemia (abnormally high levels of glucose in the blood). Most often, such insulin deficiencies result from insufficient production of islet cells in the pancreas. In most cases, this results from autoimmune destruction of these cells, a disease known as type 1 or juvenile-onset diabetes, but may also be due to physical injury or other causes.

Diabetes also results when muscle and fat cells become poorly responsive to insulin and are unable to properly absorb glucose, resulting in hyperglycemia. This phenomenon is called insulin resistance, and the resulting disease is called type 2 diabetes. Type 2 diabetes is the most common type, which is highly associated with obesity and hypertension. Obesity is associated with the inflammatory state of adipose tissue, which is thought to play a major role in the development of insulin resistance (e.g., Hotamisigil, 2006; Guilherm et al, 2008).

Diabetes is associated with damage to many tissues, primarily because hyperglycemia (and hypoglycemia, which may result from excessive or poor timed doses of insulin) is a significant source of oxidative stress. The development of chronic renal failure, retinopathy, peripheral neuropathy, peripheral vasculitis, and slow or impossible skin ulcers is among the most common complications of diabetes. Due to their ability to resist oxidative stress, in particular by inducing the expression of HO-1, the compounds of the invention can be used for the treatment of various complications of diabetes. As mentioned above (Cai et al, 2005), chronic inflammation and oxidative stress in the liver are suspected to be the major causative factors in the development of type 2 diabetes. In addition, PPAR γ agonists such as thiazolinediones are capable of reducing insulin resistance and are known to be effective therapeutic agents for type 2 diabetes.

Based on the experimental results obtained, including those given in this application, the compounds and methods of the present invention may be used to treat patients with neuroinflammation.

The therapeutic effect of diabetes can be evaluated as follows. Both the biological and, if possible, clinical efficacy of the treatment method is assessed. For example, the disease manifests itself as an increase in blood glucose, and thus the biological efficacy of the treatment can be assessed by, for example, observing the return of elevated blood glucose to normal levels. Measurement of the clinical endpoint, which may give an indication of b-cell regeneration after a period of, for example, 6-months, may give an indication of the clinical efficacy of the treatment regimen.

Based on the experimental results obtained, including those given in this application, the compounds and methods of the present invention may be used to treat diabetic patients.

G. Rheumatoid arthritis

The compounds and methods of the invention may be used to treat patients suffering from RA. Typically, the initial symptoms of Rheumatoid Arthritis (RA) occur in the inner layer of the synovium, the initial symptoms being synovial fibroblast proliferation and their attachment to the articular surface of the articular margin (Lipsky, 1998). Macrophages, T cells and other inflammatory cells are subsequently recruited into the joint where they produce a variety of mediators, including the cytokine interleukin-1 (IL-1), which promotes chronic sequelae that lead to bone and cartilage destruction, and tumor necrosis factor (TNF-. alpha.), which plays a role in inflammation (Dinarello, 1998; Arend and Dayer, 1995; van den berg, 2001). Plasma IL-1 concentrations in patients with RA are significantly higher than in healthy individuals, and it is notable that plasma IL-1 levels are associated with RA disease activity (Eastgate et al, 1988). In addition, IL-1 levels in synovial fluid are associated with various radiographic and histological features of RA (Kahle et al, 1992; Rooney et al, 1990).

In normal joints, the actions of these and other proinflammatory cytokines are balanced by a variety of anti-inflammatory cytokines and regulators (Burger and Dayer, 1995). The significance of this cytokine balance is demonstrated in juvenile RA patients who experience a periodic increase in fever throughout the day (Prieur et al, 1987). After each peak of fever, factors that block the action of IL-1 are found in serum and urine. This factor has been isolated, cloned and identified as an IL-1 receptor antagonist (IL-1ra), which is a member of the IL-1 gene family (Hannum et al, 1990). As the name suggests, IL-1ra is a natural receptor antagonist that competes with IL-1 for binding to type 1 IL-1 receptors, and as a result blocks the action of IL-1 (Arend et al, 1998). To effectively block IL-1 action, a 10 to 100-fold excess of IL-1ra is required; however, synovial cells isolated from RA patients do not appear to produce sufficient amounts of IL-1RA to counteract the effects of IL-1 (Firestein et al, 1994; Fujikawa et al, 1995).

H. Psoriatic arthritis

Psoriasis is an inflammatory and proliferative skin disease with an incidence of 1.5-3%. Approximately 20% of psoriatic patients develop a characteristic form of arthritis with several patterns (Gladman, 1992; Jones et al, 1994; Gladman et al, 1995). Some individuals present joint symptoms first, but most individuals present skin psoriasis first. Approximately one third of patients have a concurrent exacerbation of skin and interphalangeal disease (Gladman et al, 1987), and there is an anatomical correlation between the nails and distal interphalangeal arthropathy (Jones et al, 1994; Wright, 1956). Although the inflammatory process that links skin, nail and joint diseases has not yet been elucidated, immune-mediated pathologies are involved.

Psoriatic arthritis (PsA) is a chronic inflammatory joint disease characterized by a combination of arthritis and psoriasis, and was recognized in 1964 as a clinical disease distinct from Rheumatoid Arthritis (RA) (Blumberg et al, 1964). Subsequent studies have revealed that PsA shares many genetic, pathological and clinical features with other spondyloarthropathies (SpA), a group of diseases that includes ankylosing spondylitis, reactive arthritis and enteropathic arthritis (Wright, 1979). The idea that PsA belongs to the SpA group was recently further supported from imaging studies that showed that generalized osteosynthesis (enthesis) is present in PsA, but not in RA (McGonagle et al, 1999; McGonagle et al, 1998). More specifically, osteosynthesis has been postulated to be one of the earliest events occurring in SpA, leading to bone remodeling and joint stiffness in the spine, and arthromeningitis when inflamed bone sites are close to peripheral joints. However, the link between osteosynthesis and clinical presentation of PsA remains unclear, since PsA exists as a rather heterogeneous pattern of joint involvement with varying degrees of severity (Marsal et al, 1999; Salvarani et al, 1998). Therefore, other factors must be added to account for the wide variety of characteristics of PsA, only a few of which (e.g. the expression of the HLA-B27 molecule, which is highly associated with axial axis disease) are identified. As a result, it remains difficult to delineate disease signatures for specific pathogenic mechanisms, which means that treatment of the disease remains largely empirical.

Family studies have shown that genetic factors contribute to the development of PsA (Moll and Wright, 1973). Other chronic inflammatory arthritis, such as ankylosing spondylitis and rheumatoid arthritis, are considered to have a complex genetic basis. However, it is difficult to assess the genetic composition of PsA for a number of reasons. There is strong evidence that the genetic predisposition of psoriasis alone would mask genetic factors important for PsA development. Although most will receive PsA as a different disease, sometimes the phenotype overlaps with rheumatoid arthritis and ankylosing spondylitis. Likewise, PsA is not a disease of one species by itself and various subtypes have been proposed.

Increased amounts of TNF- α have been reported in psoriatic skin (Ettehadi et al, 1994) and synovial fluid (Partsch et al, 1997). Recent trials have shown that anti-TNF treatment has positive benefits in PsA (Mease et al, 2000) and ankylosing spondylitis (Brandt et al, 2000).

Based on the experimental results obtained, including those given in this application, the compounds and methods of the present invention may be used to treat psoriatic arthritis patients.

I. Reactive arthritis

In reactive arthritis (ReA), the mechanism of joint damage is unclear, but it is likely that cytokines play a key role. A more general Th1 profile has been reported with high levels of interferon gamma (IFN-. gamma.) and low levels of interleukin 4(IL-4) (Lahesmaa et al, 1992; Schlaak et al, 1992; Simon et al, 1993; Schlaak et al, 1996; Kotake et al, 1999; Ribbens et al, 2000), but several studies have shown that IL-4 and IL-10 are relatively predominant and IFN-. gamma.and tumor necrosis factor alpha (TNF-. alpha.) are relatively deficient in synovium (Simon et al, 1994; Yin et al, 1999) and Synovial Fluid (SF) (Yin et al, 1999; Yin et al, 1997) in reactive arthritis patients compared to Rheumatoid Arthritis (RA) patients. It has also been reported that TNF- α secretion levels in reactive arthritis are lower than in RA patients following ex vivo stimulation of Peripheral Blood Mononuclear Cells (PBMC) (Braun et al, 1999).

It has been discussed that clearance of reactive arthritis-associated bacteria requires the production of appropriate levels of IFN- γ and TNF- α, and that IL-10 acts by suppressing these reactions (Autenieth et al, 1994; Sieper and Braun, 1995). IL-10 is a regulatory cytokine that inhibits the synthesis of IL-12 and TNF- γ by activated macrophages (de Waal et al, 1991; Hart et al, 1995; Chomarat et al, 1995) and the synthesis of IFN- γ by T cells (Macatonia et al, 1993).

Based on the experimental results obtained, including those given in this application, the compounds and methods of the present invention may be used to treat patients with reactive arthritis.

J. Enteropathy arthritis

Typically intestinal inflammatory arthritis (EA) occurs in combination with Inflammatory Bowel Disease (IBD) such as crohn's disease or ulcerative colitis. It also affects the spine and the recommended joints of the intestine. Enteropathic arthritis involves peripheral joints, usually in the lower extremities such as the knee and ankle. It usually involves only a few or a limited number of joints and is closely associated with the bowel disease. This occurs in approximately 11% of patients with ulcerative colitis and 21% of patients with crohn's disease. Synovitis is usually self-limiting and non-deforming.

Enteropathic arthropathy comprises a group of rheumatic diseases linked to GI pathology. These diseases include reactive (i.e. infection-related) arthritis and Inflammatory Bowel Disease (IBD) associated spondyloarthropathies due to bacteria (e.g. Shigella (Shigella), Salmonella (Salmonella), Campylobacter (Campylobacter), Yersinia (Yersinia) species, Clostridium difficile), parasites (e.g. Strongyloides stercoralis, Taenia bovis (Taenia sanguinata), Giardia lamblia (Giardia lamblia), ascariasis (Ascaris lumbricoides), Cryptosporidium species). Other diseases and conditions include intestinal bypass surgery (jejunum ileum), arthritis, celiac disease, Whipple's disease, and collagenous colitis.

Based on the experimental results obtained, including those given in this application, the compounds and methods of the present invention can be used to treat patients with enteropathic arthritis.

K. Juvenile rheumatoid arthritis

Juvenile Rheumatoid Arthritis (JRA), the most prevalent form of arthritis in children, is a term applied to a family of diseases characterized by chronic inflammation and hypertrophy of the synovium. The term overlaps with, but is not fully synonymous with, a family of diseases referred to in europe as juvenile chronic arthritis and/or juvenile idiopathic arthritis.

Both the innate and adaptive immune systems use a diverse array of cell types, a large array of cell surface and secreted proteins, and an interconnected network of positive and negative feedback (Lo et al, 1999). Furthermore, although the innate and adaptive parts of the immune system are thought to be time-divisible, they cross functionally (Fearon and Locksley, 1996), and the pathological events present at these cross points may be highly correlated with our understanding of the pathogenesis of adult and childhood forms of chronic arthritis (Warrington et al, 2001).

Polyarticular JRA is a distinct clinical subtype characterized by inflammation and synovial hyperplasia in multiple joints (four or more), including the facet joints of the hand (Jarvis, 2002). This subtype of JRA can be severe because of its involvement in multiple joints and its ability to progress rapidly over time. Although clinically different, polyarticular JRA is not uniform and patients vary in disease manifestation, age of onset, prognosis and treatment response. These differences are likely to reflect a series of variations in the immune nature and inflammatory attack that can occur in the disease (Jarvis, 1998).

Based on the experimental results obtained, including those given in this application, the compounds and methods of the present invention can be used to treat JRA patients.

Early inflammatory arthritis

The compounds and methods of the present invention may be used to treat patients with early stage inflammatory arthritis. The clinical manifestations of different inflammatory arthropathies are similar in the early stages of the disease process. As a result, it is often difficult to distinguish patients who are at risk of developing severity and persistent synovitis leading to erosive joint damage from patients who are more self-limiting with respect to their arthritis. Such differentiation is crucial in order to properly target therapy, aggressively treat patients with aggressive disease and avoid unnecessary toxicity in patients with more self-limiting disease. Current clinical criteria for diagnosing erosive arthropathies, such as Rheumatoid Arthritis (RA), are poorly effective in early stages of disease, and traditional markers of disease activity, such as joint count and acute phase response, do not adequately identify patients who may have a poor prognosis (Harrison et al, 1998). The parameter that reflects the presence of a pathological event in the synovium is most likely a meaningful prognostic value.

Recent work on identifying poor prognostic predictors of early inflammatory arthritis identified the presence of RA-specific autoantibodies, in particular, the antibody anti-citrullinated peptide, associated with aggressive and persistent disease in early inflammatory arthritis-type diseases. Based on this, Cyclic Citrullinated Peptide (CCP) has been developed to help identify anti-CCP antibodies in patient sera. Using this approach, the presence of anti-CCP antibodies has been demonstrated to be RA-specific and sensitive, can distinguish RA from other arthropathies, and can effectively predict persistent, erosive synovitis before their prognosis becomes clinically manifest. Importantly, anti-CCP antibodies in serum are often detectable in many years before clinical symptoms appear, suggesting that they may reflect subclinical immune events (Nielen et al, 2004; Rantapaa-Dahlqvist et al, 2003).

Ankylosing spondylitis

AS is a subgroup of diseases within the broader disease category of spondyloarthropathies. Patients affected by multiple subgroups of spondyloarthropathies have disease etiologies in that they are often very diverse, ranging from bacterial infection to genetics. However, in all subgroups, the end result of the disease process is axial arthritis (axial arthritis). Although early clinical differences were observed in different patient populations, many of them end up with nearly identical performance after a 10-20 year course. Recent studies have shown that the mean time from onset of ankylosing spondylitis disease to clinical diagnosis is 7.5 years (Khan, 1998). These same studies indicate that spondyloarthropathy can approach the incidence of rheumatoid arthritis (Feldtkeller et al, 2003; Doran et al, 2003).

AS is a chronic systemic inflammatory rheumatic disease with or without extraosseous manifestations of the axial skeleton. The enteronomic joints and spine are mainly involved, but the hip and shoulder joints are also involved, and peripheral joints or certain extra-articular structures such as the eye, vasculature, nervous system and gastrointestinal system are often less involved. The etiology is not fully understood (Wordsworth, 1995; Calin and Taurog, 1998). It is highly associated with the major histocompatibility class I (MHC I) HLA-B27 allele (Calin and Taurog, 1998). AS affects the prime stages of an individual's life and is terrorist because it potentially causes chronic pain and irreversible damage to tendons, ligaments, joints and bones (Brewerton et al, 1973 a; Brewerton et al, 1973 b; Schlosstein et al, 1973). AS may occur alone or in combination with another form of spondyloarthropathy, such AS reactive arthritis, psoriasis, psoriatic arthritis, osteoarthritis, ulcerative colitis, irritable bowel disease or crohn's disease, in which case it is classified AS secondary AS.

Representatively, affected sites include discoverterbral joints of the spine, apophyseal joints, costovertebral and costal joints, and paraspinal ligament structures. Inflammation of the bone-knitting sites where tendons and ligaments attach to bone is also prominent in this disease (cain and Taurog, 1998). Bone site sites are known to be infiltrated by plasma cells, lymphocytes and polymorphonuclear cells. The inflammatory process often results in progressive fibrous and bony arthritic stiffness (Ball, 1971; Khan, 1990).

Diagnosis is often delayed because symptoms often present more of the more common background problems. An acute loss of lumbar flexibility is an early predictor of AS. Other common symptoms include chronic pain and stiffness in the waist, which often begins at the site where the lower spine is connected to the pelvis or hip. Although most symptoms begin in the lumbar and recommended regions of the intestine, they also involve the neck and upper back. Arthritis can also occur in the shoulders, hips and feet. Some patients have ocular inflammation and more severe patients must observe involvement of the heart valve.

The most frequent manifestation is back pain, but the disease can typically begin in peripheral joints, especially in middle-aged children and women, and rarely has acute iritis (anterior uveitis). Additional early symptoms and signs are reduced, i.e. chest fullness, low fever, fatigue, anorexia, weight loss and anemia due to diffuse rib vertebral involvement. Recurrent back pain, which is often nocturnal back pain and back pain of varying intensity, is the ultimate complaint of the disease, as morning stiffness is typically relieved by activity. A bent or bent posture relieves back pain and paraspinal muscle spasm; some degree of kyphosis is therefore common in untreated patients.

Systemic symptoms appeared in 1/3 patients. Recurrent acute iritis (anterior uveitis), which is usually self-limiting, is rarely prolonged and rarely severe enough to impair vision. Occasionally, neurological signs arise from crushed radiculitis or sciatica, spinal fractures or subluxation, and cauda equina complex (which includes impotence, nocturnal urinary incontinence, decreased bladder and rectal sensation, and lack of ankle reflexes). Cardiovascular manifestations may include aortic valve insufficiency, angina, pericarditis, and ECG conduction abnormalities. A rare finding in the lung is upper lobe fibrosis, occasionally with cavities that may be mistaken for TB and may be involved in aspergillus infection.

AS is characterized by a mild or moderately active onset of spondylitis that alternates with periods of inflammation that are nearly or completely inactive. Proper treatment in most patients results in minimal or no disability and can lead to a full life despite back stiffness. Occasionally, the course of the disease is severe and progressive, resulting in significant disability. The prognosis for patients with refractory iritis and rare patients with secondary amyloidosis is frustrating.

Based on the experimental results obtained, including those given in this application, the compounds and methods of the present invention may be used to treat patients with ankylosing spondylitis.

Ulcerative colitis

Ulcerative colitis is a disease that causes inflammation within the large intestine and sores called ulcers. Inflammation often occurs in the rectum and lower colon, but it can affect the entire colon. Ulcerative colitis affects the small intestine very little in addition to the terminal portion of the small intestine called the terminal ileum. Ulcerative colitis may also be referred to as colitis or proctitis. Inflammation often empties the colon, resulting in diarrhea. Ulceration where inflammation kills cells in the lining of the colon; ulcers lead to bleeding and suppuration.

Ulcerative colitis is an Inflammatory Bowel Disease (IBD), a common name for diseases that cause inflammation in the small intestine and colon. Ulcerative colitis is difficult to diagnose because its symptoms are similar to other bowel diseases and another type of IBD, crohn's disease. Crohn's disease differs from ulcerative colitis in that it causes deeper inflammation within the intestinal wall. Also, crohn's disease often occurs in the small intestine, although it may also occur in the mouth, esophagus, stomach, duodenum, large intestine, caecum and anus.

Ulcerative colitis may occur in people of any age, but most commonly occurs between the ages of 15 and 30, or less between the ages of 50 and 70. Sometimes children and adolescents develop the disease. Ulcerative colitis affects men and women equally and appears to occur in families. The theories as to what causes ulcerative colitis are numerous, but none have been demonstrated. The most prevalent theory is that the body's immune system reacts with viruses or bacteria through a persistent inflammatory reaction within the intestinal wall. The immune system of humans with ulcerative colitis is abnormal, but physicians are unaware of whether these abnormalities are the cause or the outcome of the disease. Ulcerative colitis is not caused by emotional distress or sensitivity to certain foods or food products, but these factors can trigger symptoms in some people.

The most common symptoms of ulcerative colitis are abdominal pain and bloody diarrhea. Patients also experience fatigue, weight loss, loss of appetite, rectal bleeding, and loss of body fluids and nutrients. About half of the patients had mild symptoms. Other patients are often with fever, bloody diarrhea, nausea and severe abdominal cramps. Ulcerative colitis may also cause problems such as arthritis, ocular inflammation, liver disease (hepatitis, cirrhosis and primary sclerosing cholangitis), osteoporosis, skin rashes and anemia. No one knows exactly why the problem occurs outside the colon. Scientists believe that these complications occur when the immune system triggers inflammation in other parts of the body. Some of these problems disappear when colitis is treated.

Diagnosis of ulcerative colitis requires a thorough physical examination and a series of tests. Blood tests can be performed to check for anemia, which would indicate bleeding in the colon or rectum. Blood tests may also reveal high white blood cell counts, which are indicative of inflammation present somewhere in the body. By examining the stool sample, the physician can detect bleeding or infection in the colon or rectum. The physician may perform a colonoscopy or sigmoidoscopy. For either test, the physician will insert an endoscope, a long, flexible, lighted tube connected to a computer and TV monitor, into the anus to view the interior of the colon and rectum. Any inflammation, bleeding or ulceration on the colon wall will have been observed. During the examination, the physician will perform a biopsy, which involves removing a tissue sample from the inner layer of the colon for microscopic observation. Barium enema x-ray examination of the colon may also be required. The method involves filling the colon with barium, a white powder solution. Barium appears white on x-ray film, which allows the physician to clearly visualize the colon, including any ulcers or other abnormalities that may be present.

Treatment of ulcerative colitis depends on the severity of the disease. Most people are treated with drugs. In severe cases, the patient will require surgery to remove the diseased colon. For ulcerative colitis, surgery is the only treatment. Some people, whose symptoms are triggered by certain foods, can control the condition by avoiding the sugars (lactose) in foods that cause bowel disorders, such as irritating foods, raw fruits and vegetables, or milk. Each one experiences ulcerative colitis will be different and therefore the treatment needs to be adjusted for each individual. Emotional and psychological support is important. Some people will relieve, i.e. the phase when symptoms disappear, which lasts for months or years. However, symptoms eventually recur in most patients. This changing pattern of disease means that people often cannot tell when treatment is helpful. Some patients with ulcerative colitis sometimes require medical care, with doctors regularly visiting to detect the condition.

Based on the experimental results obtained, including those given in this application, the compounds and methods of the present invention can be used to treat patients with ulcerative colitis.

Crohn's disease

Another condition for which immunosuppression has been attempted is crohn's disease. Symptoms of crohn's disease include intestinal inflammation and intestinal stenosis and development of intestinal fistulas; neuropathy is often accompanied by these symptoms. Anti-inflammatory drugs such as 5-aminosalicylates (e.g. mesalazine) or corticosteroids are representative prescription drugs, but they are not often effective (reviewed in Botoman et al, 1998). Immunosuppression with cyclosporine is sometimes beneficial for patients resistant or intolerant to corticosteroids (Brynskov et al, 1989).

Efforts to develop diagnostic and therapeutic tools for Crohn's disease have focused on the central role of cytokines (Schreiber, 1998; van Hogezand and Verspagent, 1998). Cytokines are small secreted proteins or factors (5 to 20kD) that have specific effects on cell-cell interactions, cell-cell communication or other cellular behaviors. The cytokines being derived from lymphocytes, particularly T_H1 and T_H2 lymphocytes, monocytes, intestinal macrophages, granulocytes, epithelial cells and fibroblasts (reviewed in Rogler and Andus, 1998; Galley and Webster, 1996). Some cytokines are pro-inflammatory (e.g., TNF-alpha, IL-1 (alpha and beta), IL-6, IL-8, IL-12, or leukocyteLeukemia Inhibitory Factor (LIF)]) (ii) a Others are anti-inflammatory (e.g., IL-1 receptor antagonists, IL-4, IL-10, IL-11, and TGF-. beta.). However, there are overlaps and functional repeats in certain inflammatory diseases.

In the case of activation of Crohn's disease, the concentration of TNF-alpha and IL-6 secreted into the blood circulation is increased, and mucosal cells overproduce TNF-alpha, IL-1, IL-6 and IL-8 locally (supra; Funakoshi et al, 1998). These cytokines can have a wide range of effects on physiological systems including bone development, hematopoiesis and liver, thyroid and neuropsychiatric functions. Also, an imbalance in the IL-1 β/IL-1ra ratio of pro-inflammatory IL-1 β predominance has been observed in Crohn's disease patients (Rogler and Andus, 1998; Saiki et al, 1998; Dionne et al, 1998; see Kuboyama, 1998). One study has shown that cytokine profiles in fecal samples would be a useful diagnostic tool for Crohn's disease (Saiki et al, 1998).

Treatments that have been proposed for Crohn's disease include the use of various cytokine antagonists (e.g., IL-1ra), inhibitors (e.g., inhibitors of IL-1. beta. converting enzyme and antioxidants), and anti-cytokine antibodies (Rogler and Andus, 1998; van Hogezand and Verspagent, 1998; Reimund et al, 1998; Lugering et al, 1998; McAllndon et al, 1998). In particular, monoclonal antibodies against TNF- α have met with some success in treating Crohn's disease (Targan et al, 1997; Stack et al, 1997; van Dullemen et al, 1995). These compounds may be used in combination therapy with the disclosed compounds.

Another approach to treating crohn's disease focuses on at least partially destroying the bacterial flora that will trigger the inflammatory response, and replacing it with a non-pathogenic flora. For example, U.S. Pat. No. 5,599,795 discloses methods for preventing and treating Crohn's disease in human patients. Their method involves sterilizing the intestinal tract with at least one antibiotic and at least one antifungal agent to kill the existing flora and replacing it with a different, selected, well characterized, bacteria obtained from normal humans. Borodys teaches a method of treating Crohn's disease by: the intestinal microflora present is at least partially removed by lavage and replaced by a new bacterial flora introduced either by fecal inoculum from a disease-free human donor or by a composition comprising Bacteroides (Bacteroides) and Escherichia coli (Escherichia coli) species (us patent 5,443,826).

Based on the experimental results obtained, including those set forth in this application, the compounds and methods of the present invention can be used to treat patients with crohn's disease.

P. systemic lupus erythematosus

The cause of autoimmune diseases such as systemic lupus erythematosus is also unknown. Systemic Lupus Erythematosus (SLE) is an autoimmune rheumatic disease characterized by the deposition of immune antibodies and immune complexes in tissues, resulting in tissue damage (Kotzin, 1996). SLE may be directly involved in multiple organ systems and the clinical manifestations are diverse and variable compared to autoimmune diseases such as MS and type 1 diabetes (reviewed in Kotzin and O' Dell, 1995). For example, some patients develop primarily rashes and joint pain, exhibit spontaneous relief and require less medication. At the other end of the range of symptoms are patients with severe and progressive renal involvement, which require treatment with high doses of steroids and cytotoxic drugs such as cyclophosphamide (Kotzin, 1996).

Serological markers and the main diagnostic test available for SLE are elevated levels of IgG antibodies against nuclear components such as double stranded DNA (dsdna), single stranded DNA (ss-DNA) and chromatin. Among these autoantibodies, the anti-dsDNA antibody IgG plays a major role in the development of lupus glomerulonephritis (G N) (Hahn and Tsao, 1993; Ohnishi et al, 1994). Glomerulonephritis is a serious disease in which the blood of the kidney purifies the capillary walls of the glomeruli to become thickened due to proliferation of the glomerular basement membrane on the epithelial side. The disease is often chronic and progressive and eventually leads to renal failure.

Based on the experimental results obtained, including those set forth in this application, the compounds and methods of the invention can be used to treat SLE patients.

Irritable bowel syndrome

The compounds and methods of the present invention may be used to treat Irritable Bowel Syndrome (IBS) patients. IBS is a functional disorder marked by abdominal pain and altered bowel habits. The syndrome may begin in young adults and may be associated with significant disability. The syndrome is not a single disorder. But rather as a subtype of IBS based on the main symptoms diarrhea, constipation or pain. In the absence of "warning" symptoms such as fever, weight loss, and gastrointestinal bleeding, limited disease detection is required. Once a diagnosis of IBS is made, the overall treatment is effective in reducing the severity of the symptoms. IBS is a common disease, although its prevalence varies. Generally speaking, IBS affects about 15% of american adults and is often about 3-fold more frequent in women than in men (Jailwala et al, 2000).

IBS results in between 240 and 350 million visits per year. Not only are the most common diseases seen by gastroenterologists, but also the most common gastrointestinal diseases seen by primary care physicians (Everhart et al, 1991; Sandler, 1990).

IBS is also a costly disease. People with IBS lose 3 times the working day and are more likely to report a failure to work due to the disease than people without bowel symptoms (Drossman et al, 1993; Drossman et al, 1997). In addition, those with IBS cost hundreds of dollars in medical costs over those without bowel disorder (Talley et al, 1995).

The absence of specific abnormalities explains the exacerbation and elimination of abdominal pain and the altered bowel habits experienced by IBS patients. The evolutionary theory of IBS suggests that there are disorders at multiple levels of the brain-gut axis. Dyskinesia, visceral hypersensitivity, abnormal regulation of the Central Nervous System (CNS) and infections are all involved. In addition, psychological factors play an important role in modification. Abnormal bowel movement has long been considered to be a factor in the pathogenesis of IBS. The transit time of the diet through the small intestine has been shown to be shorter in patients with diarrhea predominant IBS than in patients with constipation or pain predominant subtypes (Cann et al, 1983).

Both discontinuous, cluster and prolonged disseminated contractions have been reported in IBS patients in studies of the small intestine during fasting (Kellow and Phillips, 1987). They also experience pain with irregular contractions more frequently than healthy people (Kellow and Phillips, 1987; Horwitz and Fisher, 2001).

These motor findings do not account for the entire syndrome in IBS patients; in fact, most of these patients have no demonstrable abnormalities (Rothstein, 2000). IBS patients have increased sensitivity to visceral pain. Studies involving rectosigmoid sac distension have shown that IBS patients experience pain and bloating at much lower pressures and volumes than control subjects (Whitehead et al, 1990). These patients maintain normal perception of physical stimuli.

Several theories have been proposed to explain this phenomenon. For example, receptors in the gut may increase sensitivity in response to expansion or the contents of the lumen. Neurons in the dorsal horn of the spinal cord may have increased excitability. In addition, alterations in CNS sensory processes may be involved (Drossman et al, 1997). Recent functional magnetic resonance imaging studies have demonstrated that IBS patients increase activation of the anterior cingulate cortex in response to painful rectal stimulation compared to control subjects (Mertz et al, 2000).

Gradually, there is evidence of a correlation between infectious enteritis and later IBS development. Inflammatory cytokines play a role. In a survey of patients with a defined history of bacterial gastroenteritis (Neal et al, 1997), a persistent change in intestinal faecal cells was reported in 25% of patients. The persistence of symptoms results from psychological stress at the time of acute infection (Gwee et al, 1999).

Recent data indicate that bacterial overgrowth in the small intestine can play a role in IBS symptoms. In one study (Pimentel et al, 2000), 157 of the 202 IBS patients assigned to the hydrogen breathing test (78%) gave positive bacterial overgrowth. Of the 47 subjects receiving the supplemental trial, 25 subjects (53%) reported improvement with antibiotic treatment symptoms (i.e., abdominal pain and diarrhea).

IBS can present a wide range of symptoms. However, abdominal pain and altered bowel habits remain the primary features. Although the severity and location of abdominal discomfort varies greatly, abdominal discomfort is often described as spastic and located in the lower left quadrant. The patient reported diarrhea, constipation or alternating episodes of diarrhea and constipation. Symptoms of diarrhea are typically described as small loose stools, and stool is sometimes accompanied by mucous discharge. Patients also reported bloating, urgency, incomplete evacuation, and abdominal distension. Upper gastrointestinal symptoms such as gastroesophageal reflux, dyspepsia or nausea may also occur (Lynn and Friedman, 1993).

The duration of the symptoms was not an indication for further testing; it is a characteristic of IBS and is an expected symptom of its own syndrome. A more in-depth diagnostic assessment indicates a patient whose symptoms are worsening or changing. Indications for further examination also included the presence of alarm symptoms, onset of symptoms after age 50 and a family history of colon cancer. The tests may include colonoscopy, computed tomography imaging of the abdomen and pelvis, and barium studies of the small or large intestine.

R. sicca syndrome

The compounds and methods of the invention may be used to treat SS patients. Primary Sjogren's Syndrome (SS) is a chronic, slowly progressive systemic autoimmune disease that affects primarily middle-aged women (female to male ratio 9: 1), but can be observed in all ages, including children (Jonsson et al, 2002). It is characterized by lymphocytic infiltration and destruction of the exocrine glands by monocytes, including CD4+, CD8+ lymphocytes and B cells (Jonsson et al, 2002). Furthermore, extraglandular (systemic) manifestations were observed in 1/3 patients (Jonsson et al, 2001).

Lymphocyte infiltration of the gland is a progressive feature (Jonsson et al, 1993) that displaces a large proportion of the organs when extensively infiltrated. Interestingly, in some patients, the glandular infiltration in the salivary glands closely resembles the ectopic lymphocyte microstructure (named germinal center) (Salomonsson et al, 2002; Xanthou et al, 2001). In SS, ectopic GC is defined as a T cell and B cell mass with follicular dendritic cells and proliferating cells that activate the endothelial cell network. These GC-like structures formed in the target tissues also characterize the functional properties of generating autoantibodies (anti-Ro/SSA and anti-La/SSB) (Salomonson and Jonsson, 2003).

In other systemic autoimmune diseases, such as RA, factors that are critical for GC have been identified. Rheumatoid synovial tissue with GC was shown to produce the chemokines CXCL13, CCL21 and Lymphotoxin (LT) - β (detected in follicular center and mantle B cells). Multivariate regression analysis of these analytes identified CXCL13 and LT- β as the only cytokines predictive of GC in rheumatoid synovitis (Weyand and Goronzy, 2003). It has recently been demonstrated that CXCL13 and CXCR5 in salivary glands play an essential role in inflammatory processes by recruiting B and T cells, thus promoting lymphocyte regeneration and ectopic GC formation in SS (salomonson et al, 2002).

Psoriasis S

The compounds and methods of the present invention may be used to treat patients with psoriasis. Psoriasis is a chronic skin peeling and inflammatory disease that affects 2% to 2.6% of americans, or between 580 and 750 million people. Although the disease occurs in all age groups, it primarily affects adults. Roughly equal between men and women. Psoriasis occurs when skin cells rise rapidly from a source below the skin surface and accumulate on the surface before they have a chance to mature. Typically, this movement (also called renewal) takes about 1 month, but in psoriasis it occurs only within a few days. In its typical form, psoriasis results in a thick red (inflamed) skin patch overcoated with silvery scales. These patches, sometimes referred to as plaques, are often itchy or painful. They most often occur on the elbows, knees, other parts of the legs, scalp, lower back, face, palms, and soles of the feet, but they also occur on the skin of any part of the body. The disease can also affect the nails, toenails, and genitals and soft tissues within the mouth. Although the skin around the affected joint is usually cracked open, approximately 100 million patients with psoriasis experience inflammation of the joint, producing arthritic symptoms. The disease is called psoriatic arthritis.

Psoriasis is an immune system, and in particular relates to immune system-driven skin disorders of the type known as T-cell leukocytes. Normally, T cells help protect the body against infection and disease. In the case of psoriasis, T cells function incorrectly and become so active that they trigger other immune responses, which lead to inflammation and rapid renewal of skin cells. In about 1/3 cases, there is a family history of psoriasis. Researchers have studied a large number of psoriasis-affected families and identified genes associated with the disease. People with psoriasis notice the time for their skin to deteriorate and then improve. Conditions that lead to redness include infection, stress, and climate change that dries the skin. Likewise, some medications, including hypertensive medications lithium and beta blockers, can trigger outbreaks or exacerbations of the disease.

Infectious diseases

The compounds of the present disclosure may be used to treat infectious diseases, including viral infections and bacterial infections. As mentioned above, such infections may be associated with severe local or systemic inflammatory responses. For example, influenza can produce severe pneumonia, and bacterial infections can lead to high systemic inflammatory responses, including overproduction of diverse inflammatory cytokines, which are hallmarks of sepsis. In addition, the compounds of the present invention may be used to directly inhibit the replication of viral pathogens. Previous studies have demonstrated that related compounds such as CDDO can inhibit HIV replication in macrophages (vazzez et al, 2005). Other studies have shown that inhibition of the NF-. kappa.B signaling pathway inhibits replication of influenza virus, and that cyclopentenone prostaglandins inhibit viral replication (e.g., Mazur et al, 2007; Pica et al, 2000).

Pharmaceutical formulations and routes of administration

The compounds of the present disclosure can be administered by a variety of methods, such as orally or by injection (e.g., subcutaneously, intravenously, intraperitoneally, etc.). Depending on the route of administration, the active compound may be coated in a material to protect the compound from the action of acids and other natural conditions that would inactivate the compound. They may also be administered by continuous perfusion/perfusion at the site of disease or trauma.

In order to administer therapeutic compounds by means other than parenteral administration, it is necessary to coat the compounds or co-administered compounds with a material that protects them from inactivation. For example, the therapeutic compound may be administered to the patient in a suitable carrier such as a liposome or diluent. Pharmaceutically acceptable diluents include salts and aqueous buffers. Liposomes include water-in-oil-in-water CGF emulsions as well as conventional liposomes (Strejan et al, 1984).

The therapeutic compounds may also be administered parenterally, intraperitoneally, intraspinally, or intracerebrally. Dispersions can be prepared in glycerol, liquid polyethylene glycols and mixtures thereof and in oils. Under normal conditions of storage and use, these formulations will contain a preservative to prevent microbial growth.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (when water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the composition must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion vehicle including, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersants, and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be better to include isotonic agents, for example, sugars, sodium chloride or polyalcohols such as mannitol and sorbitol in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.

Sterile injectable solutions can be prepared by incorporating the therapeutic compound in the required amount in the appropriate solvent with one or a combination of the ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the therapeutic compound into a sterile vehicle which contains the basic dispersion vehicle and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient (i.e., the therapeutic compound) plus any additional desired ingredient from a previously sterile-filtered solution thereof.

The therapeutic compound may be administered orally, for example, with an inert diluent or an assimilable edible carrier. The therapeutic compound and other ingredients may also be enclosed in hard or soft shell gelatin capsules, compressed in tablets or incorporated directly into the subject's food. For oral therapeutic administration, the therapeutic compound may be admixed with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, cachets and the like. Of course, the percentage of therapeutic compound in the compositions and formulations can vary. The amount of therapeutic compound in such therapeutically useful compositions is such that a suitable dosage will be obtained.

It is particularly advantageous to formulate parenteral compositions in unit dosage form for ease of administration and uniformity of dosage. As used herein, dosage form refers to physically discrete units suitable as unitary dosages for the subjects to be treated; each unit containing a predetermined quantity of therapeutic compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for the dosage unit forms of the invention are determined by and directly depend upon (a) the unique characteristics of the therapeutic compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such therapeutic compounds for the treatment of selected patient conditions.

The therapeutic compounds may also be administered topically to the skin, eye or mucosa. Alternatively, if local delivery to the lung is desired, the therapeutic compound may be administered by inhalation of a dry powder or aerosol formulation.

The active compound is administered in a therapeutically effective amount sufficient to treat a condition associated with the condition in the patient. A "therapeutically effective amount" preferably reduces the total number of disease symptoms in an infected patient by at least about 20%, more preferably by at least about 40%, even more preferably by at least about 60%, and still more preferably by at least about 80% compared to an untreated subject. For example, the efficacy of a compound can be assessed in an animal model system, such as the model systems shown in the examples and figures, which predicts the efficacy of treatment of a human disease.

The actual dose of a compound of the present disclosure or a composition comprising a compound of the present disclosure administered to a subject may be determined by physical and physiological factors such as age, sex, body weight, severity of the disease, type of disease to be treated, previous or concurrent therapeutic measures, specific symptoms of the subject, and route of administration. These factors can be determined by the skilled artisan. The practitioner responsible for administration will typically determine the concentration of one or more active ingredients in the composition and one or more appropriate dosages for the individual subject. The dosage can be adjusted by the individual physician in the event of any complications.

An effective amount will typically vary from about 0.001mg/kg to about 1000mg/kg, from about 0.01mg/kg to about 750mg/kg, from about 100mg/kg to about 500mg/kg, from about 1.0mg/kg to about 250mg/kg, from about 10.0mg/kg to about 150mg/kg, administered in one or more doses per day for one or more days (depending, of course, on the mode of administration and the factors described above). Other suitable dosage ranges include 1mg to 10,000 mg/day, 100mg to 10000 mg/day, 500mg to 10000 mg/day, and 500mg to 1000 mg/day. In some particular embodiments, the amount is less than 10,000 mg/day, with an amplitude ranging from 750mg to 9,000 mg/day.

The effective amount may be less than 1 mg/kg/day, less than 500 mg/kg/day, less than 250 mg/kg/day, less than 100 mg/kg/day, less than 50 mg/kg/day, less than 25 mg/kg/day, or less than 10 mg/kg/day. Alternatively, it is in the range of 1 mg/kg/day to 200 mg/kg/day. For example, for the treatment of a diabetic patient, the dosage unit may be an amount that reduces blood glucose by at least 40% compared to an untreated subject. In another embodiment, the unit dose is an amount that reduces blood glucose to a level of ± 10% of the blood glucose level of the non-diabetic subject.

In other non-limiting examples, the dosage may also include a dosage selected from the group consisting of about 1 μ g/kg/body weight, about 5 μ g/kg/body weight, about 10 μ g/kg/body weight, about 50 μ g/kg/body weight, about 100 μ g/kg/body weight, about 200 μ g/kg/body weight, about 350 μ g/kg/body weight, about 500 μ g/kg/body weight, about 1 mg/kg/body weight, about 5 mg/kg/body weight, about 10 mg/kg/body weight, about 50 mg/kg/body weight, about 100 mg/kg/body weight, about 200 mg/kg/body weight, about 350 mg/kg/body weight, about 500 mg/kg/body weight to about 1,000 mg/kg/body weight or higher, any range derivable therein. In non-limiting examples of ranges derived from the values listed above, ranges of about 5 mg/kg/body weight to about 100 mg/kg/body weight, about 5 μ g/kg/body weight to about 500 mg/kg/body weight, and the like, may be administered based on the above values.

In certain embodiments, a pharmaceutical composition of the present disclosure may comprise, for example, at least about 0.1% of a compound of the present disclosure. In further embodiments, the compounds of the present disclosure may comprise between about 2% to about 75% weight units or between about 25% to about 60% weight units, and any range derivable therein.

Single or multiple doses of the agent are contemplated. The desired time interval for multiple dose delivery can be determined by one of ordinary skill in the art, using no more than routine experimentation. For example, a subject may be administered doses twice daily at approximately 12 hour intervals. In some embodiments, the agent is administered once daily.

The one or more agents may be administered on a conventional schedule. As used herein, a conventional schedule refers to a predetermined period of time. The regular schedule may include time periods that are the same or different in length, so long as the schedule is preset. For example, a conventional schedule may include administration twice daily, once every two days, once every three days, once every four days, once every five days, once every six days, once weekly, once monthly, or any set number of days or weeks therebetween. In other embodiments, the invention provides that one or more agents may be orally ingested and with a time frame that is dependent or independent of food intake. Thus, for example, the agent may be ingested daily in the morning and/or daily in the evening, regardless of when the subject is or will be eating.

Combination therapy

In addition to use as monotherapeutic agents, the compounds of the present disclosure may also be used in combination therapy. Effective combination therapy can be achieved using a single composition or pharmaceutical formulation comprising two agents, or using two different compositions or formulations simultaneously, wherein one composition comprises a compound of the disclosure and the other comprises one or more second agents. Alternatively, the treatment may precede or follow treatment with other agents at intervals ranging from minutes to months.

Various combinations may be employed, for example, when a compound of the present disclosure is "a" and "B" represents a second agent, non-limiting examples of which are described below:

A/B/A B/A/B B/B/A A/A/B A/B/B B/A/A A/B/B/B B/A/B/B

B/B/B/A B/B/A/B A/A/B/B A/B/A/B A/B/B/A B/B/A/A

B/A/B/A B/A/A/B A/A/A/B B/A/A/A A/B/A/A A/A/B/A

administration of the compounds of the present disclosure to patients will follow the general protocol for drug administration, taking into account the toxicity of the drug if present. It is contemplated that the treatment cycle will be repeated as needed.

Interferon beta will be a suitable second agent. There are human cytokine-derived drugs that help modulate the immune system. They include interferon beta-1 b and interferon beta-1 a. Betaseron has been approved by the FDA for relapsing secondary progressive MS. In addition, the FDA has approved several interferon betas for use as therapeutic agents to treat patients who have experienced a single episode that has been demonstrated to be multiple sclerosis and patients who may be at risk for further episodes and develop clear MS. For example, when an MRI scan of the brain shows a high risk of damage that predicts a shift to clear MS, a risk of MS is indicated.

Glatiramer acetate is another example of a second agent that may be used in combination therapy. Glatiramer is currently used to treat relapsing remitting MS. It consists of four amino acids found in myelin. The drug reportedly stimulates T cells in the immune response of the body to change from a harmful pro-inflammatory agent to a beneficial anti-inflammatory agent that reduces inflammation at the site of injury.

Another potential second agent is mitoxantrone, a chemotherapeutic drug for many cancers. The drug is also FDA approved for the treatment of aggressive forms of relapsing-remitting MS, as well as certain forms of progressive MS. Typically, it is administered intravenously every three months. The drug is effective, but is limited by cardiotoxicity. Norstrin has been approved by the FDA for secondary progressive, progressive-relapsing, and exacerbation relapsing-remitting MS.

Another potential second agent is natalizumab. In general, natalizumab acts by blocking the attachment of immune cells to the brain blood vessels, which is an essential step in the entry of immune cells into the brain, thus reducing the inflammatory effects of immune cells on brain neurons. Natalizumab has been shown to significantly reduce the frequency of episodes in patients with relapsing MS.

In the case of relapsing-remitting MS, a corticosteroid, such as methylprednisolone, may be administered intravenously to the patient as a second agent to more quickly terminate the challenge and leave less persistent deficiency.

Other commonly used drugs for MS that may be used in combination with the compounds of the present disclosure include immunosuppressive drugs such as azathioprine, cladribine, and cyclophosphamide.

It is contemplated that other anti-inflammatory agents may be used in combination with the therapeutic agents of the present invention. Other COX inhibitors may be used, including arylcarboxylic acids (salicylic acid, acetylsalicylic acid, diflunisal, choline magnesium trisalicylate, salicylates, paracetamol, flufenamic acid, mefenamic acid, meclofenamic acid, and triflumic acid), arylalkanoic acids (diclofenac, fenclofenac, fentiazac, ibuprofen, flurbiprofen, ketoprofen, naproxen, fenoprofen, fenbufen, suprofen, indoprofen, tiaprofenic acid, benzofenamic acid, and the like), arylalkanoic acids (diclofenac, fenclofenac, fentiaprofenic acid, and the like)Loxifene, pirprofen, tolmetin, zomepirac, clininac, indomethacin, and sulindac) and enolic acids (phenylbutazone, oxybutyzone, apazone, feprazone, piroxicam, and isoxicam. See, for example, U.S. patent 6,025,395.

Histamine H2 receptor blocking agents may also be used in combination with the compounds of the present invention, including cimetidine, ranitidine, famotidine and nizatidine.

Acetylcholinesterase inhibitor therapies, such as tacrine, donepezil, metrazoxane, and rivastigmine, for the treatment of alzheimer's disease and other diseases are also contemplated for use in combination with the disclosed compounds. Other acetylcholinesterase inhibitors developed that can be used once approved include rivastigmine and metrazine. Acetylcholinesterase inhibitors increase the amount of neurotransmitter acetylcholine at nerve terminals by reducing the destruction of acetylcholine by cholinesterase.

MAO-B inhibitors such as selegiline may be used in combination with the compounds of the present invention. Selegiline is used in parkinson's disease and irreversibly inhibits type B monoamine oxidase (MAO-B). Monoamine oxidase is an enzyme that inactivates the monoamine neurotransmitters norepinephrine, serotonin and dopamine.

Food and nutritional supplements that are reported to be beneficial for the treatment or prevention of parkinson's disease, alzheimer's disease, multiple sclerosis, amyotrophic lateral sclerosis, rheumatoid arthritis, inflammatory bowel disease and all other diseases whose pathogenesis is believed to involve overproduction of any of Nitric Oxide (NO) or prostaglandins, for example acetyl-L-carnitine, octacosanol, evening primrose oil, vitamin B6, tyrosine, phenylalanine, vitamin C, L-dopa or several antioxidant combinations may be used in combination with the compounds of the present invention.

For the treatment or prevention of cancer, the compounds of the present invention may be used in combination with one or more of the following therapeutic agents: radiation, chemotherapeutic agents (e.g., cytotoxic agents such as anthracyclines, vincristine, vinblastine, agents targeting microtubules such as paclitaxel and docetaxel, 5-FU and related agents, cisplatin and other platinum-containing compounds, irinotecan and topotecan, gemcitabine, temozolomide, etc.), targeted therapeutic agents (e.g., imatinib, bortezomib, bevacizumab, rituximab), or vaccine therapeutic agents designed to promote enhanced immune responses targeting cancer cells.

For the treatment or prevention of autoimmune diseases, the compounds of the present invention may be used in combination with one or more of the following drugs: corticosteroids, methotrexate, anti-TNF antibodies, other TNF-targeted protein therapeutics and NSAIDs. For the treatment or prevention of cardiovascular diseases, the compounds of the present invention may be used in combination with antithrombotic therapeutic agents, anticholesterol therapeutic agents such as statins (e.g. atorvastatin) and surgical interventions such as stents or coronary artery bypass grafts. For the treatment of osteoporosis, the compounds of the invention may be used in combination with anti-resorptive agents such as the bisphosphonates or anabolic therapeutic agents such as teriparatide or parathyroid hormone. For the treatment of neuropsychiatric disorders, the compounds of the present invention may be used in combination with antidepressants (e.g., imipramine or SSRI such as fluoxetine), antipsychotics (e.g., olanzapine, sertindole, risperidone), mood stabilizers (e.g., lithium, half sodium valproate), or other standard agents such as anxiolytic agents. For the treatment of neurological disorders, the compounds of the invention may be used in combination with anticonvulsants (e.g. monosodium valproate, gabapentin, phenytoin, carbamazepine and topiramate), antithrombotic agents (e.g. tissue plasminogen activator) or analgesics (e.g. opiates, sodium channel blockers and other anti-pain-sensing agents).

Examples

The following examples are included to illustrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques disclosed by the inventors to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

Example 1 methods and materials

Nitric oxide production and cell viability. RAW264.7 macrophages were pretreated with DMSO or drug for 2 hours and then treated with recombinant mouse IFN γ (Sigma) for 24 hours. The NO concentration in the medium was determined using the Griess reagent System (Promega). Cell viability was determined using WST-1 reagent (Roche).

iNOS induced qPCR. RAW264.7 mouse macrophages were pretreated with the indicated concentrations of the compounds for 2 hours and then stimulated with 10ng/ml IFN γ for an additional 2 hours. mRNA levels of iNOS were quantified by qPCR and the values for the vehicle treated IFN γ stimulated samples were normalized to a value of 1, relative to the display of the vehicle treated IFN γ stimulated samples. Values are the average of PCR reactions in duplicate and triplicate wells.

iNOS induced Western blotting. RAW264.7 cells were pretreated with the indicated compounds for 2 hours, then stimulated with 10ng/ml IFN γ for an additional 24 hours. Levels of iNOS protein were determined by immunoblotting. Actin was used as loading control.

The compounds were compared. In some experiments (e.g., fig. 17, 18), compounds of the invention were compared to other synthetic triterpenoids and natural products, such as those shown here:

compound 402 can be prepared according to the methods taught by Honda et al, (1998), Honda et al, (2000b), Honda et al, (2002), Yates et al, (2007), and U.S. patent nos. 6,326,507 and 6,974,801, which are incorporated herein by reference.

Example 2 Synthesis of certain Natural products containing an anti-inflammatory pharmacophore

Curcumin analogs C0008, C0009 and C0010 were synthesized as described in schemes 1-3 below.

Reagents and conditions for scheme 1: (a) at 130 ℃, 16h, 68 percent; (b) (i) LHMDS, MeI, -78 ℃ to room temperature, 3 h; (ii)LHMDS, MeI, -78 ℃ to room temperature, 3h, 71%; (c) i is₂，50℃，24h，95％；(d)NaBH₄，CeCl·7H₂O, 0 ℃, 1.5h, 80%; (e) TMSCl, imidazole, 0 ℃, 1h, 95%; (f) (i) DIBAL-H, -78 ℃, 1H; (ii) DMP, room temperature, 20min, 85%.

Compound 3 (68%) was prepared by a Diels-Alder reaction of compounds 1 and 2 as shown in scheme 1 using a modification of the method reported by Danishefsky et al, 1979. After introduction of the gem-dimethyl to give compound 4 (71% yield), iodoethylene 5 (95% yield) was produced. The enone was reduced to allylic alcohol 6 and protected to give TMS ether 7 (80% yield for both steps). The methyl ester 7 was then converted to the aldehyde 9 in two steps (91% yield).

Reagents and conditions for scheme 2: (a) LDA, 70 percent at the temperature of-78 ℃ to 0 ℃ for 2.5 h; (b) et3N, MsCl, 0 ℃, 30min, 81%; (c) (i) ethylene glycol, TsOH, 110 ℃, 4 h; (ii) TBSCl, imidazole, room temperature 43%; (d) BuLi, dimethyl methylphosphonate, 91% at-78 ℃ to room temperature.

Compound 12 (70%) is obtained from the aldol condensation of ketone 10 and aldehyde 11 (Cardona et al, 1986) (scheme 2). Compound 12 was converted to the mesylate, which was removed in situ to give compound 13 (81% yield). After protection of the ketene to the ketal (43%), the methyl ester was converted to dimethyl phosphate 15 in excellent yield (91%).

Compound 15 and compound 9 obtained above were condensed and converted into the target curcuminoid analog (scheme 3 below). The Horner-Wadsworth-Emmons reaction between phosphate 15 and aldehyde 9 was first attempted using NaHMDS as the base. No reaction was observed, most likely due to steric hindrance of 9. The reaction proceeded slowly to give compound 16 in 28% yield using the protocol developed by Roush et al, 1984 (DIPEA/LiCl). The iodoethylene 16 was then passed through Pd (PPh)₃)₄Catalytic Zn (CN)₂Treatment (Wu et al, 1999) and yield cyanide 17 (68%). After removal of the TMS protecting group and oxidation, compound C0008 was obtained (76% yield from 17).

C0008 is the starting material that yields both C0009 and C0010 (scheme 3 below). When C0008 was treated with 6N HCl (aq), the fully deprotected compound C0009 was obtained (88%). When C0008 was treated with TBAF, phenol C0010 was obtained in 63% yield.

Reagents and conditions for scheme 3: (a) DIPEA, LiCl, 60 ℃, 20h, 28%; (b) zn (CN)₂，Pd(PPh₃)₄68% at 80 ℃ for 20 min; (c) TsOH, room temperature, 30min, 99%; (d) DMP, room temperature, 1h, 77%; (e)6N HCl (aq), room temperature, 14h, 88% (for C0009); TBAF, room temperature, 10min, 63% (for C0010).

Scheme 4

Reagents and conditions for scheme 4: (a) (i) K₂CO₃Room temperature, 2 h; (ii) MOMCl, DIPEA, room temperature, 14h, 94%.

The synthesis of resveratrol analog R00141 is summarized in scheme 4 and scheme 5 below. Starting material 3 is first treated with I₂the/Py treatment yielded iodoethylene 21 (74%). The iodoethylene is then protected to give the ketal 22, which is used directly in the next reaction. Ketal 22 is converted to aldehyde 24 in two steps and the yield is high (91%). Reaction of aldehyde 24 with diethyl phosphate 20 under basic conditions yielded compound 25 in 89% yield. Iodoethylene 25 is represented by Zn (CN)₂And a catalytic amount of Pd (PPh)₃)₄The cyanide 26 was obtained in 62% yield. In that The ketal and MOM protecting groups were removed in one step using TsOH to afford R00141 (87%).

Scheme 5

Reagents and conditions for scheme 5: (a) i is₂Room temperature, 16h, 74%; (b) ethylene glycol, 110 ℃, 3 h; (c) (i) DIBAL-H, -78 ℃, 1H; (ii) DMP, room temperature, 20min, 91%; (d) t-BuOK, 0 ℃ to room temperature, 2h, 89%; (e) zn (CN)₂，Pd(PPh₃)₄62% at 80 ℃ for 20 min; (f) TsOH, room temperature, 2min, 87%.

Scheme 6

Reagents and conditions for scheme 6: (a) TsOH, room temperature, 89%; (b) (i) LHMDS, MeI, -78 ℃ to room temperature, 3 h; (ii) LHMDS, MeI, 48% at-78 ℃ to room temperature, 3 h; (b) zn (CN)₂，Pd(PPh₃)₄42% at 80 ℃ for 20 min; (f) TsOH, room temperature, 2min, 93%.

R00142 was synthesized from compound 25 (scheme 6). Compound 25 is treated with TsOH to selectively remove the ketal group to give enone 27 (89%). Then the gem-dimethyl was introduced using LHMDS/MeI. Compound 28 was isolated from the reaction mixture in 48% yield. A mixture of compounds 28 and 29 was also obtained (14%). Using the same protocol as described for the synthesis of R00141, R00142 was obtained from compound 28 in 39% yield. From the mixture of compounds 28 and 29, a mixture of R00142 and compound 31 (designated R00142-1) was obtained.

Reagents and conditions for scheme 7: (a) t-BuOK, MeI, Room temperature, 4h, 62%; (b) h2, 10% Pd/C, room temperature, 16H, 38%; (c) imidazole, TMSCl, room temperature, 1h, 84%; (d) (i) LDA, TsCN, -78 ℃ to 0 ℃ for 1 h; (ii) DDQ, 80 ℃, 20min, 50%; (e) TsOH, room temperature, 5min, 95%; (f) DMP, room temperature, 2h, 79%.

DHEA analog D0016 is synthesized from testosterone (scheme 7). Testosterone treatment with t-BuOK/MeI (Cao et al, 2007) introduced a gem-dimethyl group at position 4, giving compound 32 in 62% yield. Compound 32 is then hydrogenated to yield 33 (38%). After the 17-hydroxy group was protected (84% yield), compound 34 was treated with LDA/TsCN (Kahne and Collum, 1981) and then subjected to DDQ oxidation to give the desired compound D0016 in 50% yield.

D0017 and D0018 were synthesized from D0016 (scheme 7). Treatment of D0016 with TsOH gave D0017 in 95% yield. Oxidation of D0017 with Dess-Martin oxidant (Dess-Martin periodinane) gave compound D0018 (79%).

Reagents and conditions for scheme 8: (a) MOMCl, DIPEA, DMAP, room temperature, 14h, 98%; (b) h₂O₂NaOH, 4 ℃, 14h, 71%; (c) NaCN, EtOH, 80 ℃, 24 h; (d) at 210 ℃ for 40min, a yield of 23% D0014 (36) and a yield of 23% D0015 (36).

Compound D0014 was prepared from compound 35 using the protocol reported by Rasmusson et al, 1986 (scheme 8). Compound 35 (produced in 98% yield from testosterone) was treated with alkaline hydrogen peroxide to give a mixture of epoxide epimers. The mixture was converted to the alkali soluble dicyano compound 37, which was used directly in the next step. Mild pyrolysis of compound 37 yielded the corresponding 4-cyanosteroids D0014 (23%) and D0015 (23%).

Reagents and conditions for scheme 9: (a) (i) K₂CO₃Room temperature, 4 h; (ii) PCC, NaOAc, room temperature, 3h, 95%; (b) (i) LDA, TsCN, -78 ℃ to 0 ℃ for 1 h; (ii) DDQ, 80 ℃, 20min, 37%.

The nucleochrysin analog H0001 was synthesized from compound 38 (scheme 9). Compound 38 was prepared from erythrogenin acetate using the reported method (Barton et al, 1980). Acetate 38 was treated with base and then subjected to PCC oxidation to afford ketone 39 in 95% yield. Then 39 was reacted with LDA/TsCN (Kahne and Collum, 1981), followed by DDQ oxidation to give the desired compound H0001 (37%).

Scheme 10:

reagents and conditions for scheme 10: (a) ethylene glycol, CSA, cyclohexylamine, Dean-Stark, refluxing for 20h, 99%; (b) 3-methyl-2-butanone, Al (Oi-Pr)₃Toluene, reflux, 4h, 72%; (c) KOt-Bu (1M in THF), t-BuOH, allyl bromide, room temperature, 2h, 73%; (d) LDA, THF, at-78 deg.C for 30 min; TsCN, 57% at-78 deg.C for 30 min; (e) h ₂(1atm), 10% Pd/C, THF, 2h, 100%; (f) (i) DDQ, benzene, 80 ℃, 3 h; (ii)1N HCl (aq), THF, RT, 2h, 10%.

Reagents and conditions for scheme 11: (a) grubbs catalyst (2 nd generation), CH₂Cl₂Room temperature, 2h, 99%; (b) LDA, THF, toluene, 30min at-78 deg.C; TsCN, -734% at 8 ℃ for 30 min; (c) DDQ, benzene, 80 ℃, 30min, 18%; (d)1N HCl (aq), THF, H₂O, room temperature, 14h, 64%.

Reagents and conditions for scheme 12: (a) h₂(1atm), 5% Pd/C, THF, 2h, 98%; (b) LDA, THF, at-78 deg.C for 30 min; TsCN, 55% at-78 deg.C for 30 min; (c) (i)1, 3-dibromo-5, 5-dimethylhydantoin, DMF, 0 ℃, 2 h; (ii) pyridine, 55 ℃, 21h, 76% (d)0.5N HCl (aq), THF, room temperature, 50h, 98%; (e) al (Oi-Pr)₃i-PrOH, toluene, 75 ℃, 20h, 20%.

Reagents and conditions for schemes 13a and 13 b: (a) AcCl, MeOH, room temperature, 72 h; (b) ag₂CO₃Kieselguhr, toluene, reflux, 3h, 66%; (c) MOM-Cl, i-Pr₂Net，CH₂Cl₂，45℃，14h，70％；(d)NaOMe，HCO₂Et, MeOH, rt, 2 h; (e) NH (NH)₂OH-HCl，EtOH，H₂O, 60 ℃, 14h, 40%; (f) NaOMe, MeOH, THF, 55 ℃, 2h, 41%; (g) (i)1, 3-dibromo-5, 5-dimethylhydantoin, DMF, room temperature, 3 h; (ii) pyridine, 55 ℃, 72h, 80%; (h)2N HCl (Et) ₂O)，CH₂Cl₂Room temperature, 14h, 48%.

Example 3-characterization of certain Natural products including anti-inflammatory pharmacophores

Compound (I)3: a mixture of compound 1(4.20g, 24.4mmol) and 2(2.44g, 24.4mmol) in toluene (1mL) was heated in a sealed tube at 130 ℃ for 16 h. After cooling to room temperature, THF (10mL) and 0.1N HCl (5mL) were added. After stirring for 20min, NaHCO was added₃(aq) solution, and the mixture was extracted with EtOAc. The combined extracts were washed with water and MgSO₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 15% EtOAc in hexanes) to give compound 3 as a colorless oil (2.84g, 68%).¹H NMR(400MHz，CDCl₃) δ 6.89(d, 1H, J ═ 10.4Hz), 5.98(d, 1H, J ═ 10.4Hz), 3.75(s, 3H), 2.50(m, 3H), 1.99(m, 1H), 1.45(s, 3H). The 1H NMR spectrum was identical to that reported in the literature (Danishefsky et al, 1979).

Compound 4: LHMDS (1.0M in THF, 3.75mL, 3.75mmol) was added dropwise to a solution of compound 3(505mg, 3.00mmol) in THF (20mL) at-78 deg.C. After stirring for 1h, MeI (0.56mL, 9.00mmol) was added and the reaction mixture was stirred at room temperature for 2 h. After cooling to 0 ℃, NH was added₄Cl solution, and the mixture with CH ₂Cl₂And (4) extracting. The combined extracts were washed with water and MgSO₄Dried and concentrated. The resulting crude product was dissolved in THF (20mL) and cooled to-78 ℃. LHMDS (1.0M in THF, 3.75mL, 3.75mmol) was added again dropwise. After stirring at-78 ℃ for 1h, MeI (0.56mL, 9.00mmol) was added and the reaction mixture was stirred at room temperature for 2 h. After cooling to 0 ℃, NH was added₄Cl solution, and the mixture with CH₂Cl₂And (4) extracting. The combined extracts were washed with water and MgSO₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 10% EtOAc in hexanes) to give compound 4(420mg, 71%) as a white solid:¹H NMR(300MHz，CDCl₃)δ6.78(dd，1H，J＝1.2，10.2Hz)，5.91(d，1H，J＝10.2Hz)，3.74(s，3H)，2.54(dd，1H，J＝1.2，14.1Hz)，1.79(d，1H，J＝14.1Hz)，1.40(s，3H)，1.12(s，3H)，1.02(s，3H)。

compound 5: mixing Compound 4(212mg, 1.08mmol), I₂(824mg，3.24mmol) and pyridine (1.5mL) in CCl₄The mixture in (3mL) was heated at 50 ℃ for 24 h. After cooling to room temperature, EtOAc (20mL) was added. The mixture is mixed with Na₂S₂O₃The solution, 1N HCl (aq) and water were washed, then MgSO₄And (5) drying. After concentration, the resulting residue was purified by column chromatography (silica gel, 7% to 10% EtOAc in hexanes) to give compound 5(313mg, 95%).¹H NMR(400MHz，CDCl₃)δ7.59(s，1H)，3.76(s，3H)，2.59(d，1H，J＝14.4Hz)，1.87(d，1H，J＝14.4Hz)，1.42(s，3H)，1.19(s，3H)，1.05(s，3H)；¹³C NMR(100MHz，CDCl₃)δ25.0，26.0，27.9，41.4，45.5，46.9，52.7，103.0，157.7，174.6，196.2。

Compound 6: to a solution of compound 5(200mg, 0.62mmol) in MeOH (6mL) at 0 deg.C was added CeCl₃·7H₂O (255mg, 0.68mmol) and NaBH ₄(26mg, 0.68 mmol). After stirring for 1h, NaBH was added again₄(26mg, 0.68mmol) and stirred for another 30 min. Water was added to the reaction mixture, and the mixture was extracted with EtOAc. The combined extracts were washed with water and MgSO₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 10% to 20% EtOAc in hexanes) to afford allylic alcohol 6(160mg, 80%):¹HNMR(400MHz，CDCl₃)δ6.51(s，1H)，3.77(dd，1H，J＝1.2，6.0Hz)，3.69(s，3H)，2.34(d，1H，J＝14.4Hz)，2.00(d，1H，J＝6.0Hz)，1.42(d，1H，J＝14.0Hz)，1.26(s，3H)，1.07(s，3H)，0.84(s，3H)；¹³C NMR(100MHz，CDCl₃)δ19.1，28.0，28.6，36.0，44.5，46.7，52.3，78.2，107.3，141.2，175.8。

compounds 8 and 9: imidazole (203mg, 2.99mmol) and TMSCl (190. mu.L, 1.49mmol) were added successively to alcohol 6(160mg, 0.50mmol) in CH at 0 deg.C₂Cl₂(3 mL). After stirring for 1h, NaHCO was added₃(aq) solution, and the mixture with CH₂Cl₂And (4) extracting. The mixed extract is extracted with MgSO₄Drying and concentration gave Compound 7(194mg, 95%). Compound 7 was used in the next step without further purification.

DIBAL-H (1.0M in toluene, 0.50mL, 0.50mmol) was added to compound 7(194mg, 0.47mmol) in CH at-78 deg.C₂Cl₂(5 mL). After stirring for 30min, DIBAL-H (1.0M in toluene, 0.50mL, 0.50mmol) was added again and stirred for another 30 min. A solution of potassium sodium tartrate (aq) was added and the mixture was stirred at room temperature until clear. Using it with CH₂Cl₂Extracting, and washing the combined extracts with water and MgSO ₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 10% EtOAc in hexanes) to give alcohol 8(150mg, 87%) and aldehyde 9(11mg, 6.4%). Compound 8: 1HNMR (300MHz, CDCl)₃)δ6.09(s，1H)，3.80(s，1H)，3.21(m，2H)，2.19(t，1H，J＝5.4Hz)，1.88(d，1H，J＝14.4Hz)，1.07(d，1H，J＝14.4Hz)，0.99(s，3H)，0.98(s，3H)，0.91(s，3H)，0.20(s，9H)；¹³C NMR(100MHz，CDCl₃)δ0.90，24.8，26.3，27.7，37.0，37.6，43.0，71.2，83.0，100.5，144.2。

Will CH₂Cl₂NaHCO in (2mL)₃(430mg, 5.12mmol) and dess-Martin oxidant (434mg, 1.02mmol) were stirred at room temperature for 20min, after which alcohol 8(150mg, 0.41mmol) in CH was added₂Cl₂(20 mL). After stirring for 2h, Na was added₂S₂O₃(aq) solution and stirred for 10 min. The mixture is extracted with hexane and the combined extracts are extracted with NaHCO₃(aq) extraction with MgSO 2₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 5% EtOAc in hexanes) to afford aldehyde 9(128mg, 85%):¹H NMR(300MHz，CDCl₃)δ9.37(s，1H)，6.29(s，1H)，3.89(s，1H)，2.11(d，1H，J＝14.0Hz)，1.29(d，1H，J＝14.0Hz)，1.10(s，3H)，1.01(s，3H)，0.83(s，3H)，0.23(s，9H)。

compound 12: n-BuLi (2.5M in hexane, 1.46mL, 3.65mmol) was added to diisopropylamine (0.54mL, 3.82mmol) in THF (2mL) at-78 ℃. In thatAfter stirring at 0C for 30min, it was cooled again to-78 ℃. Compound 10(500mg, 3.47mmol) in THF (5mL) was then added dropwise. After stirring for 1h, Compound 11(Cardona et al, 1986) (1.11g, 4.17mmol) was added and the resulting mixture was stirred at-78 ℃ for 2 h. Addition of NH₄Cl (aq) the reaction was stopped and the mixture was extracted with ether. The combined extracts were washed with water and MgSO ₄And (5) drying. After concentration, the resulting residue was purified by column chromatography (silica gel, 10% to 20% EtOAc in hexanes) to afford product 12(1.0g, 70%):¹H NMR(400MHz，CDCl₃)δ6.73-6.89(m，3H)，5.10(ddd，1H，J＝3.2，6.4，8.4Hz)，3.81(s，3H)，3.72(s，3H)，3.20(d，1H，J＝2.8Hz)，2.88(dd，1H，J＝8.8，17.6Hz)，2.80(dd，1H，J＝3.6，17.6Hz)，1.37(s，3H)，1.35(s，3H)，0.99(s，9H)，0.14(s，6H)。

compound 13: et at 0 ℃₃N (1.06mL, 7.62mmol) and MsCl (0.24mL, 3.09mmol) were added sequentially to compound 12(1.00g, 2.42mmol) in CH₂Cl₂(20 mL). After stirring for 30min, NaHCO was added₃(aq), and the mixture was extracted with EtOAc. The combined extracts were washed with water and MgSO₄And (5) drying. After concentration, the resulting residue was purified by column chromatography (silica gel, 10% EtOAc in hexanes) to give product 13(0.78g, 81%): 1H NMR (400MHz, CDCl)₃)δ7.64(d，1H，J＝16.0Hz)，7.06(m，1H)，6.99(bs，1H)，6.83(d，1H，J＝8.0Hz)，6.63(d，1H，J＝16.0Hz)，3.84(s，3H)，3.71(s，3H)，1.43(s，6H)，0.98(s，9H)，0.16(s，6H)。

Compound 14: a mixture of compound 13(3.70g, 9.44mmol), ethylene glycol (6.0mL), and TsOH (0.80g) in toluene (25mL) was refluxed with a Dean-Stark apparatus for 4 h. After cooling to room temperature, the mixture was washed with NaHCO₃(aq) solution and water, then MgSO₄And (5) drying. After concentration, the brown oil was dissolved in DMF (5 mL). Imidazole (2.70g, 39.7mmol) and TBSCl (1.50g, 10mmol) were added. After stirring for 30min, TBSCl (500mg, 3.33mmol) was added again and stirred for another 30 min. Then NaHCO is added₃(aq) solution and the mixture was extracted with EtOAc. The combined extracts were washed with water and MgSO ₄And (5) drying. After concentration, the resulting residue was purified by column chromatography (silica gel, 10% to 15% EtOAc in hexanes) to afford product 14(1.71g, 43%):¹H NMR(400MHz，CDCl₃)δ6.85-6.90(m，2H)，6.79(d，1H，J＝8.4Hz)，6.60(d，1H，J＝16.0Hz)，6.07(d，1H，J＝16.0Hz)，3.95(m，4H)，3.83(s，3H)，3.70(s，3H)，1.30(s，6H)，1.00(s，9H)，0.15(s，6H)。

compound 15: n-BuLi (2.5M in hexane, 3.30mL, 8.25mmol) was added to dimethyl methylphosphonate (1.0mL, 9.35mmol) in THF (10mL) at-78 ℃. After stirring for 15min, a solution of compound 14(1.20g, 2.75mmol) in THF (5mL) was added dropwise. After stirring at room temperature for 2h, NH was added₄Cl (aq) solution. The mixture was extracted with EtOAc and the combined extracts were washed with water and MgSO₄And (5) drying. After concentration, the resulting residue was purified by column chromatography (silica gel, 50% to 100% EtOAc in hexanes) to afford product 14(1.32g, 91%):¹HNMR(400MHz，CDCl₃)δ6.84-6.89(d，2H)，6.79(d，1H，J＝8.0Hz)，5.57(d，1H，J＝16.0Hz)，5.87(d，1H，J＝16.0Hz)，3.90-4.01(m，4H)，3.82(s，3H)，3.80(s，3H)，3.77(s，3H)，3.47(d，2H，J＝20.4Hz)，1.26(s，6H)，0.99(s，9H)，0.15(s，6H)。

compound 16: the condensation reaction between compounds 9 and 15 was carried out using the method developed by Roush et al, 1984. MeCN (10mL) and DIPEA (1.25mL, 7.18mmol) were added to a mixture of LiCl (121mg, 2.85mmol) and compound 15(830mg, 1.71mmol) to give a yellow solution. Compound 9(520mg, 1.42mmol) in MeCN (5mL) was then added. The resulting reaction mixture was heated at 60 ℃ for 20h and cooled to room temperature. EtOAc was added and the mixture was washed with water, NaHCO₃(aq) solution and brine, then MgSO ₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 6% to 7% EtOAc in hexanes) to give product 16(310 mg). The recovered starting materials 9 and 15 were again treated with the same reaction conditions to yield another 285mgProduct 16 of (a). The total yield is 55%:¹H NMR(400MHz，CDCl₃)δ6.82-6.86(m，2H)，6.77(d，1H，J＝8.0Hz)，6.70(bs，2H)，6.55(d，1H，J＝15.6Hz)，6.28(s，1H)，5.88(d，1H，J＝15.6Hz)，3.90(m，4H)，3.84(bs，1H)，3.81(s，3H)，1.80(d，1H，J＝14.8Hz)，1.40(d，1H，J＝14.8Hz)，1.22(s，6H)，1.11(s，3H)，0.99(s，3H)，0.98(s，9H)，0.83(s，3H)，0.23(s，9H)，0.14(s，6H)；¹³C NMR(100MHz，CDCl₃)δ-4.4，1.4，18.7，20.7，24.1，25.9，28.8，28.9，37.7，42.9，44.9，54.1，55.7，64.9，65.0，81.5，105.4，110.3，111.4，120.4，121.1，124.1，124.2，130.1，132.2，143.3，145.5，151.2，152.7，201.7。

compound 17: pd (PPh) in DMF (10mL) at room temperature under Ar₃)₄(90mg, 0.078mmol) was added to compound 16(595mg, 0.77mmol) and Zn (CN)₂(363mg, 3.10mmol) in the mixture. The slurry was heated at 80 ℃ for 20min and cooled to room temperature. Ether was added and the mixture was washed with water, MgSO₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 5% to 10% EtOAc in hexanes) to afford product 17(352mg, 68%):¹H NMR(400MHz，CDCl₃)δ6.82-6.85(m，2H)，6.79(d，1H，J＝8.0Hz)，6.68(m，2H)，6.55(d，1H，J＝16.0Hz)，6.51(bs，1H)，5.86(d，1H，J＝16.0Hz)，3.90(m，5H)，3.82(s，3H)，1.80(d，1H，J＝14.4Hz)，1.42(d，1H，J＝14.0Hz)，1.23(s，3H)，1.23(s，3H)，1.15(s，3H)，0.99(s，9H)，0.95(s，3H)，0.78(s，3H)，0.23(s，9H)，0.15(s，6H)。

compound 18: TsOH (240mg, 1.26mmol) was added to a solution of compound 17(160mg, 0.24mmol) in acetone (5mL) and water (1mL) at room temperature. After stirring for 30min, NaHCO was added₃(aq), and the mixture was extracted with EtOAc. The combined extracts were washed with water and MgSO₄And (5) drying. After concentration, the resulting residue was purified by column chromatography (silica gel, 20% to 33% EtOAc in hexanes) to afford product 18(142mg, 99%):¹H NMR(400MHz，CDCl₃) δ 6.82-6.85(m, 2H), 6.79(d, 1H, J ═ 8.0Hz), 6.77(d, 1H, J ═ 16.0Hz), 6.66(d, 1H, J ═ 16.0Hz), 6.59(m, 1H), 6.55(d, 1H, J ═ 15.6Hz), 5.85(d, 1H, J ═ 15.6Hz), 3.96(dd, 1H, J ═ 1.6, 6.8Hz), 3.92(m, 4H), 3.82(s, 3H), 1.82(d, 1H, J ═ 14.4Hz), 1.48(d, 1H, J ═ 14.0Hz), 1.24(s, 3H), 1.23(s, 3H), 1.15(s, 3H), 1.03(s, 3H), 3.99 (s, 0, 9H), 9.80 (s, 0H), 9H, 0 s, 9H. The compound was contaminated with some unidentified impurities.

Compound C0008: c0008(110mg, 77%) was generated strictly from compound 18(142mg, 0.24mmol) using the method described for the synthesis of compound 9 from compound 8:¹H NMR(300MHz，CDCl₃)δ7.50(d，1H，J＝1.5Hz)，6.76-6.83(m，4H)，6.72(d，1H，J＝16.2Hz)，6.53(d，1H，J＝15.9Hz)，5.82(d，1H，J＝15.9Hz)，3.91(m，4H)，3.81(s，3H)，2.06(dd，1H，J＝1.5，14.7Hz)，1.95(d，1H，J＝14.4Hz)，1.33(s，3H)，1.24(s，3H)，1.24(s，3H)，1.16(s，3H)，1.05(s，3H)，0.98(s，9H)，0.14(s，6H)；m/z 616.3(M+Na⁺)。

compound C0009: a mixture of C0008(90mg, 0.15mmol), 6N HCl (aq) (1mL), and THF (5mL) was stirred at room temperature overnight. EtOAc is added and the mixture is washed with water, MgSO₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 33% EtOAc in hexanes) to give product C0009(58mg, 88%):¹H NMR(300MHz，CDCl₃)δ7.64(d，1H，J＝15.3Hz)，7.39(d，1H，J＝1.5Hz)，7.10(dd，1H，J＝1.8，8.4Hz)，6.89-6.98(m，3H)，6.56(d，1H，J＝15.3Hz)，6.12(d，1H，J＝15.3Hz)，5.93(s，1H)，3.94(s，3H)，2.03(dd，1H，J＝1.8，14.7Hz)，1.92(d，1H，J＝14.4Hz)，1.42(s，3H)，1.41(s，3H)，1.33(s，3H)，1.11(s，3H)，0.92(s，3H)；¹³C NMR(100MHz，CDCl₃)δ20.8，20.8，25.8，25.9，29.2，39.4，41.5，47.0，56.0，60.4，110.3，114.0，114.9，115.7，118.3，123.4，124.0，126.4，145.2，146.8，148.7，151.1，162.5，196.2，197.4，197.6；m/z 458.1(M+Na⁺)。

compound C0010: TBAF (1.0M in THF, 74. mu.L, 0.074mmol) was added to a solution of C0008(40mg, 0.067mmol) in THF (3 mL). After stirring for 10min, EtOAc was added. The mixture was washed with water and MgSO₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 33% EtOAc in hexanes) to give product C0010(20mg, 63%):¹H NMR(400MHz，CDCl₃)7.49(d，1H，J＝1.6Hz)，6.86(m，3H)，6.84(d，1H，J＝16.0Hz)，6.72(d，1H，J＝16.0Hz)，6.53(d，1H，J＝16.0Hz)，5.82(d，1H，J＝16.0Hz)，5.71(s，1H)，3.92(bs，7H)，2.06(dd，1H，J＝2.0，14.4Hz)，1.95(d，1H，J＝14.4Hz)，1.34(s，3H)，1.25(s，3H)，1.24(s，3H)，1.17(s，3H)，1.06(s，3H)；m/z 480.2(M+1)。

compound 20: k₂CO₃A mixture of (7.00g, 50.7mmol) and compound 19(2.70g, 9.44mmol) in MeOH (150mL) was stirred at room temperature for 2 h. The mixture was filtered and the resulting filtrate was concentrated to give the crude potassium salt, which was suspended in THF (50 mL). DIPEA (5.42mL, 31.2mmol) and MOMCl (2.70mL, 35.5mmol) were added sequentially. The white slurry was stirred at room temperature overnight. EtOAc is added and the mixture is washed with water, MgSO ₄Dried and concentrated. The oil obtained is purified by column chromatography (silica gel, CH)₂Cl₂Medium 5% MeOH) to give product 20(2.53g, 94%).

Compound 21: compound 3(280mg, 1.67mmol), I₂(630mg, 2.48mmol) and pyridine (2mL) in CCl₄The mixture in (4mL) was stirred at room temperature for 16 h. EtOAc was added and the mixture was taken over Na₂S₂O₃(aq), 1N HCl (aq) and water, then MgSO₄And (5) drying. After concentration, the resulting residue was purified by column chromatography (silica gel, 12% EtOAc in hexanes) to give compound 21(364mg, 74%):¹H NMR(300MHz，CDCl₃)7.68(s，1H)，3.77(s，3H)，2.69-2.74(m，2H)，2.52(m，1H)，2.04(m，1H)，1.47(s，3H)；¹³C NMR(75MHz，CDCl₃)δ24.7，32.5，33.4，48.0，52.8，104.3，159.6，173.3，191.1。

compound 22: a mixture of compound 21(370mg, 1.26mmol), ethylene glycol (1mL), PPTS (80mg) in toluene (3mL) was heated at reflux with a Dean-Stark apparatus for 3 h. After cooling to room temperature, EtOAc was added. NaHCO for the mixture₃(aq) solution, water wash, and MgSO₄And (5) drying. After concentration, the crude product 22(400mg, 94%) was obtained, which was used in the next step without further purification. Compound 22:¹H NMR(300MHz，CDCl₃)6.65(s，1H)，4.21(m，2H)，3.98(m，2H)，3.69(s，3H)，2.30(m，1H)，1.96(m，2H)，1.75(m，1H)，1.30(s，3H)。

compound 23, 24: using the procedure described for the synthesis of compounds 8 and 9 from compound 7, compounds 23(185mg, 50%) and 24(150mg, 41%) were generated from compound 22(400mg, 1.18 mmol).

Compound 24(158mg, 85%) was produced from compound 23(185mg, 0.60mmol) using the procedure described for the synthesis of compound 9 from compound 8.

Compound 23:¹H NMR(400MHz，CDCl₃)6.45(s，1H)，4.21(m，2H)，3.98(m，2H)，3.39(m，2H)，1.85-2.00(m，4H)，1.54(m，1H)，1.04(s，3H)；¹³C NMR(100MHz，CDCl₃)δ22.2，28.7，30.5，42.4，65.4，65.6，69.7，104.5，105.8，149.6。

compound 24:¹H NMR(400MHz，CDCl₃)9.43(s，1H)，6.47(s，1H)，4.21(m，2H)，3.98(m，2H)，2.16(m，1H)，1.94(m，2H)，1.71(m，1H)，1.16(s，3H)；¹³C NMR(100MHz，CDCl₃)δ20.8，27.3，30.6，52.1，65.7，65.8，105.1，107.4，143.4，199.9。

compound 25: potassium tert-butoxide (790mg, 7.05mmol) in THF (20mL) was added to a solution of compound 20(2.13g, 7.39mmol) in THF (10mL) at 0 deg.C. After stirring for 45min, a solution of compound 24(512mg, 1.67mmol) in THF (10mL) was added. The mixture was stirred at 0 ℃ for 1h and at room temperature for a further 1 h. EtOAc is added and the mixture is washed with water and MgSO₄And (5) drying. After concentration, the resulting residue was purified by column chromatography (silica gel, 12% EtOAc in hexanes) to give the product 25(680mg, 89%):¹H NMR(400MHz，CDCl₃)7.27(d，2H，J＝10.4Hz)，6.97(d，2H，J＝10.4Hz)，6.47(s，1H)，6.27(d，1H，J＝16.4Hz)，5.92(d，1H，J＝16.0Hz)，5.16(s，2H)，4.22(m，2H)，3.97(m，2H)，3.48(s，3H)，1.94(m，2H)，1.80(m，2H)，1.12(s，3H)；NMR(100MHz，CDCl₃)δ27.2，30.6，33.3，43.0，55.9，65.4，65.8，94.3，103.4，105.9，116.2，127.3，128.0，130.9，133.7，150.4，156.6。

compound 26: compound 26(36mg, 62%) was produced from compound 25(75mg, 0.17mmol) using the procedure described for the synthesis of compound 17 from compound 16.¹H NMR(400MHz，CDCl₃)7.28(d，2H，J＝10.4Hz)，6.98(d，2H，J＝10.4Hz)，6.54(s，1H)，6.21(d，1H，J＝16.4Hz)，5.91(d，1H，J＝16.0Hz)，5.17(s，2H)，4.24(m，2H)，3.96-4.07(m，2H)，3.47(s，3H)，1.78-1.89(m，4H)，1.27(s，3H)。

Compound R00141: TsOH (77mg, 0.40mmol) was added to a solution of compound 26(36mg, 0.081mmol) in acetone (3mL) at room temperature and the acetone was removed under reduced pressure to give a white solid which was dried in vacuo (2mm Hg) for 2 min. Acetone (10mL) and NaHCO were added₃(1.0 g). After stirring for 5min, the mixture was filtered through a pad of celite. The filtrate was concentrated and the crude product was purified by column chromatography (silica gel, 15% to 25% EtOAc in hexanes) to give the product R00141(18mg, 87%): ¹H NMR(400MHz，CDCl₃)7.47(d，1H，J＝1.2Hz)，7.25(d，2H，J＝9.2Hz)，6.82(d，2H，J＝8.8Hz)，6.27(d，1H，J＝16.4Hz)，5.96(d，1H，J＝16.4Hz)，5.26(bs，1H)，2.52-2.66(m，2H)，2.04-2.14(m，2H)，1.43(s，3H)；¹³C NMR(100MHz，CDCl₃)δ26.8，33.6，34.2，40.0，114.0，115.7，116.4，127.8，128.6，129.2，130.5，155.8，167.6，192.1。

Compound 27: TsOH (5.04g, 26.5mmol) was added to a solution of compound 25(680mg, 1.50mmol) in acetone (25mL) and water (5 mL). The mixture was stirred at room temperature until complete consumption of compound 25 was confirmed by TLC analysis. EtOAc was added and the mixture was washed with water, NaHCO₃(aq) solution washing and MgSO 2₄And (5) drying. After concentration, the resulting residue was purified by column chromatography (silica gel, 10% EtOAc in hexanes) to give the product 25(550mg, 89%):¹H NMR(300MHz，CDCl₃)7.52(s，1H)，7.28(m，2H)，7.00(m，2H)，6.33(d，1H，J＝16.2Hz)，5.98(d，1H，J＝16.2Hz)，5.18(s，2H)，3.48(s，3H)，2.68(m，2H)，2.09(m，2H)，1.37(s，3H)。

compound 28: compound 28(280mg, 48%) was produced from compound 27(550mg, 1.34mmol) using the procedure described for the synthesis of compound 4 from compound 3. From this reaction, a mixture of compounds 28 and 29 was obtained (85mg, 14%).

Compound 28:¹H NMR(400MHz，CDCl₃)7.61(s，1H)，7.28(d，2H，J＝8.8Hz)，6.99(d，2H，J＝8.8Hz)，6.29(d，1H，J＝16.4Hz)，6.07(d，1H，J＝16.4Hz)，5.17(s，2H)，3.47(s，3H)，2.05(dd，1H，J＝1.2，14.4Hz)，1.99(d，1H，J＝14.0Hz)，1.32(s，3H)，1.20(s，3H)，1.16(s，3H)；¹³C NMR(100MHz，CDCl₃)δ27.1，27.8，29.9，42.0，43.2，48.6，55.9，94.3，102.8，116.3，127.2，128.0，130.5，134.8，156.8，162.6，197.2。

compound 30: using the procedure described for the synthesis of compound 17 from compound 16, compound 30(90mg, 42%) was generated from compound 28(280mg, 0.66 mmol):¹H NMR(400MHz，CDCl₃)7.56(d，1H，J＝1.6Hz)，7.28(m，2H)，7.01(m，2H)，6.26(d，1H，J＝16.4Hz)，6.07(d，1H，J＝16.4Hz)，5.18(s，2H)，3.47(s，3H)，2.07(dd，1H，J＝1.6，14.8Hz)，1.96(d，1H，J＝14.8Hz)，1.39(s，3H)，1.20(s，3H)，1.17(s，3H)；¹³C NMR(100MHz，CDCl₃)δ26.3，26.5，29.7，39.4，41.7，47.8，56.0，94.2，114.5，114.9，116.4，127.3，128.9，130.0，133.3，157.1，165.7，197.2。

compound C00142: compound C00142(70mg, 93%) was generated from compound 30(90mg, 0.27mmol) using the procedure described for the synthesis of C00141 from compound 26:¹H NMR(300MHz，CDCl₃)7.56(d，1H，J＝1.5Hz)，7.21(m，2H)，6.83(m，2H)，6.23(d，1H，J＝16.5Hz)，6.07(bs，1H)，6.01(d，1H，J＝16.5Hz)，2.04(dd，1H，J＝1.5，14.7Hz)，1.94(d，1H，J＝14.7Hz)，1.37(s，3H)，1.19(s，3H)，1.16(s，3H)；¹³C NMR(75MHz，CDCl₃)δ26.3，26.5，29.6，39.4，41.7，47.7，114.5，114.7，115.7，127.5，128.7，129.1，132.5，155.9，166.4，197.7；m/e 282.1(M+1)。

r00142-1: c00142-1 (a 1: 1 mixture of 31 and C00142) was prepared from a mixture of compounds 28 and 29 (2mg, 4.6. mu. mol) using the method described for the synthesis of C00142 from compound 28. The 1H NMR of identified compound 31 is: (400MHz, CDCl) ₃)7.41(d, 1H, J ═ 2.4Hz), 6.23(d, 1H, J ═ 16.4Hz), 5.97(d, 1H, J ═ 16.4Hz), 2.61(m, 1H), 1.39(s, 3H), 1.15(d, 3H, J ═ 6.8 Hz); LC-MS showed that the retention times of compound 31 and C00142 were 6.52min, M/z 290.0(M + Na), respectively⁺) And 7.36min, M/z 304.0(M + Na)⁺)。

Compound 32: testosterone (5.19g, 18.0mmol) was added in one portion to a solution of t-BuOK (5.96g, 53.2mmol) in t-BuOH (100mL) at room temperature. After stirring for 5min, MeI (6.64mL, 106.4mmol) was added dropwise over 10 min. After stirring at room temperature for 4h, water (75mL) was added and t-BuOH was removed under reduced pressure. The precipitated white solid was collected by filtration and washed with water. White solid dissolved in CH₂Cl₂In (1), with MgSO₄Dried and concentrated. The crude product was recrystallized from acetone (80mL) to give compound 32(3.50g, 62%) as a white crystalline solid:¹H NMR(400MHz，CDCl₃)δ5.56(dd，1H，J＝2.8，6.8Hz)，3.66(m，1H)，2.40-2.62(m，2H)，1.99-2.19(m，3H)，1.86(m，1H)，1.26-1.70(m，9H)，1.24(s，6H)，0.94-1.18(m，3H)，0.87(s，3H)，0.77(s，3H)；¹³C NMR(75MHz，CDCl₃)δ11.0，19.3，20.8，23.3，27.2，30.2，30.5，31.2，31.2，32.0，33.6，36.5，37.1，42.7，48.6，49.0，51.3，81.7，119.6，149.8，216.7。

compound 33: compound 32(1.05g, 3.32mmol) in EtOH (50mL) and 10% Pd on carbon (500mg) were hydrogenated at room temperature (1atm) for 16 h. The reaction mixture was filtered through a pad of celite, and the filtrate was concentrated to give the crude product, which was purified by column chromatography (silica gel, 5% to 9% CH in EtOAc)₂Cl₂) Compound 33(400mg, 38%) was obtained as a white solid: ¹H NMR(300MHz，CDCl₃)δ3.63(m，1H)，2.63(ddd，1H，J＝6.6，12.9，15.5Hz)，2.32(ddd，1H，J＝3.3，5.4，15.5Hz)，1.92-2.14(m，2H)，1.77-1.83(m，2H)，1.19-1.65(m，11H)，1.08(m，1H)，1.06(s，6H)，1.05(s，3H)，0.65-0.95(m，3H)，0.75(s，3H)；¹³C NMR(75MHz，CDCl₃)δ11.1，14.0，20.4，21.8，22.3，23.3，25.7，30.5，32.0，34.6，35.2，36.5，36.5，38.1，42.9，47.8，50.9，55.5，55.8，81.8，217.4。

Compound 34: imidazole (125mg, 1.84mmol) and TMSCl (155. mu.L, 1.22mmol) were added to compound 33(195mg, 0.61mmol) in CH at room temperature₂Cl₂(6.0 mL). After stirring for 1h, the reaction mixture was washed with NaHCO₃(aq) washing with MgSO 2₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 5% hexane in ether) to give compound 34(200mg, 84%):¹HNMR(400MHz，CDCl₃)δ3.53(dd，1H，J＝8.0，8.4Hz)，2.63(ddd，1H，J＝7.2，13.2，15.2Hz)，2.32(ddd，1H，J＝3.2，5.2，15.2Hz)，1.97(ddd，1H，J＝3.2，5.6，13.2Hz)，1.71-1.91(m，3H)，1.19-1.59(m，10H)，1.06(s，3H)，1.05(s，6H)，0.78-0.99(m，3H)，0.70(s，3H)，0.66(m，1H)，0.07(s，9H)；¹³C NMR(75MHz，CDCl₃)δ0.1，11.3，13.9，20.5，21.7，22.3，23.4，25.7，30.8，32.1，34.7，35.2，36.5，36.8，38.1，42.9，47.8，50.6，55.6，56.0，81.6，217.3。

compound D0016: n-BuLi (2.5M in hexane, 0.41mL, 1.03mmol) was added to diisopropylamine (159. mu.L, 1.12mmol) in THF (0.6mL) at-78 ℃.After stirring at 0 ℃ for 30min, the reaction mixture was cooled again to-78 ℃ and compound 34(200mg, 0.51mmol) in THF (2mL) was added dropwise. After stirring at-78 ℃ for 30min, TsCN (371mg, 2.04mmol) in THF (2.0mL) was added. After stirring for another 30min, the reaction was quenched by addition of water (1 mL). The reaction mixture was adjusted to pH 3 using 3N HCl (aq) and extracted with EtOAc. The combined extracts were washed with water and MgSO₄Dried and concentrated. The resulting crude product was dissolved in benzene (15mL) and DDQ (116mg, 0.51mmol) was added. The red solution was refluxed for 20min and cooled to room temperature. NaHCO for reaction mixture₃(aq) solution and water, then MgSO ₄And (5) drying. After concentration, the residue obtained is purified by column chromatography (silica gel, CH)₂Cl₂Medium 0 to 5% EtOAc) to give D0016(106mg, 50%):¹H NMR(400MHz，CDCl₃)δ7.84(s，1H)，3.55(dd，1H，J＝8.4，8.4Hz)，1.80-1.92(m，3H)，1.64-1.73(m，3H)，1.40-1.58(m，5H)，1.26(m，1H)，1.19(s，3H)，1.17(s，3H)，1.11(s，3H)，0.86-1.16(m，4H)，0.73(s，3H)，0.08(s，9H)。

compound D0017: p-TsOH (220mg, 1.16mmol) was added to a solution of D0016(95mg, 0.23mmol) in acetone (2mL) and water (0.4 mL). After stirring at room temperature for 5min, NaHCO was added₃(aq) solution, and the mixture with CH₂Cl₂And (4) extracting. The mixed extract is extracted with MgSO₄Dried and concentrated. The resulting crude product was purified by column chromatography (silica gel, 33% EtOAc in hexanes) to afford D0017(75mg, 95%):¹H NMR(300MHz，CDCl₃)δ7.85(s，1H)，3.67(dd，1H，J＝8.4，8.7Hz)，2.08(m，1H)，1.20-1.96(m，12H)，1.19(s，3H)，1.18(s，3H)，1.11(s，3H)，0.86-1.14(m，4H)，0.78(s，3H)；¹³C NMR(75MHz，CDCl₃)δ11.5，16.1，20.7，21.6，22.0，23.5，27.0，30.6，31.5，35.6，36.5，40.8，43.2，45.1，51.1，51.3，51.9，81.6，114.7，115.3，168.8，198.2。

compound D0018: NaHCO is added at room temperature₃(140mg, 1.66mmol) and dess-Martin oxidant (177mg, 0.42mmol) in the solvent phaseFollowed by addition of D0017(57mg, 0.17mmol) in CH₂Cl₂(3 mL). After stirring for 2h, 5% Na was added₂S₂O₃(aq) solution. The reaction mixture was extracted with ether and the combined extracts were extracted with NaHCO₃(aq) washing with MgSO 2₄Dried and concentrated. The resulting crude product was purified by column chromatography (silica gel, 20% EtOAc in hexanes) to give D0018(45mg, 79%) which was contaminated with some impurities. Recrystallization from EtOAc/hexanes yielded D0018(38mg) as a purified white solid:¹H NMR(300MHz，CDCl₃)δ7.84(s，3H)，2.48(m，1H)，1.26-2.18(m，12H)，1.21(s，3H)，1.20(s，3H)，1.13(s，3H)，1.08(m，1H)，0.91(s，3H)；¹³C NMR(75MHz，CDCl₃)δ13.8，15.8，20.1，21.2，21.6，21.7，26.7，30.5，31.0，34.8，35.6，40.5，44.8，47.5，50.9，51.1，51.5，114.6，114.8，167.8，197.7，219.9。

compound 35: diisopropylethylamine (12.2mL, 70.0mmol), MOMCl (2.66mL, 35.0mmol) and DMAP (0.21g, 1.7mmol) were added sequentially to testosterone (5.05g, 17.5mmol) in CH ₂Cl₂(50mL) in a stirred solution. After stirring at room temperature for 14h, NaHCO was added₃(aq) solution. After stirring for 10min, the organic layer was separated and washed with 1N HCl (aq), NaHCO₃(aq) solution and water, then MgSO₄And (5) drying. After concentration, the resulting residue was purified by column chromatography (silica gel, 9% to 33% EtOAc in hexanes) to afford compound 35(5.66g, 98%) as a white solid:¹H NMR(400MHz，CDCl₃)δ5.73(bs，1H)，4.63(m，2H)，3.53(dd，1H，J＝8.4，8.4Hz)，3.35(s，3H)，2.32-2.47(m，3H)，2.28(m，1H)，2.04(m，2H)，1.90(m，1H)，1.85(m，1H)，1.24-1.75(m，7H)，1.19(s，3H)，1.15(m，1H)，0.88-1.06(m，3H)，0.82(s，3H)。

compounds D0014 and D0015: a solution of compound 35(1.66g, 5.0mmol) in MeOH (80mL) at 0 deg.C with 30% H₂O₂(aq) (3.52mL, 35.2mmol) and NaOH (aq) (2.5N, 1.40mL, 3.50 mmol). Stirring at 4 deg.C for 14h, addingWater (200mL) was added and the reaction mixture was extracted with EtOAc. The combined organic layers were washed with water and MgSO₄Drying and concentration gave epoxide 36(1.24g, 71%) as an epimer mixture. A solution of epoxide 36(1.20g, 3.47mmol) in EtOH (60mL) was treated with a solution of NaCN (1.70g, 34.7mmol) in water (18 mL). After refluxing for 24h, EtOH was removed by evaporation. A 10% naoh (aq) solution (30mL) was added at room temperature and the mixture was extracted with ether. The aqueous layer was acidified to pH2 with 6N HCl (aq) (30mL) at 0 ℃ and extracted with EtOAc. The combined EtOAc extracts were washed with water and MgSO ₄Drying and concentration gave crude product 37 as a white foamy solid. Compound 37 was heated at 210 ℃ under vacuum (5mmHg) for 40min and cooled to room temperature. The brown viscous oil was purified by column chromatography (silica gel, 33% to 50% EtOAc in hexanes) to give compound D0014(290mg, 23% from 36) and D0015(260mg, 23% from 36) as white solids.

D0014：¹H NMR(400MHz，CDCl₃)δ4.63(m，2H)，3.54(dd，1H，J＝8.4Hz)，3.35(s，3H)，3.07(ddd，1H，J＝2.4，3.6，15.2Hz)，1.90-2.13(m，4H)，1.43-1.80(m，7H)，1.30-1.40(m，2H)，1.27(s，3H)，1.10-1.21(m，2H)，0.94-1.10(m，2H)，0.83(s，3H)；¹³C NMR(100MHz，CDCl₃)δ11.6，18.0，20.5，23.1，27.9，31.1，32.0，33.0，34.0，34.9，36.6，40.2，42.4，50.1，53.7，55.1，86.0，95.9，112.1，114.0，184.0，192.3；m/z 358.2(M+1)。

D0015：¹H NMR(300MHz，CDCl₃)δ3.67(dd，1H，J＝8.4Hz)，3.07(ddd，1H，J＝2.7，3.9，15.3Hz)，2.40-2.56(m，3H)，1.96-2.16(m，3H)，1.89(ddd，1H，J＝3.0，3.6，12.6Hz)，1.57-1.79(m，5H)，1.43-1.55(m，2H)，1.34(m，1H)，1.28(s，3H)，0.80-1.20(m，4H)，0.80(s，3H)；¹³C NMR(75MHz，CDCl₃)δ11.0，18.0，20.6，23.2，30.3，31.2，32.0，33.0，34.0，35.1，36.1，40.3，42.7，50.1，53.7，81.3，112.1，114.0，184.1，192.4；m/z 314.1(M+1)。

Compound 39: compound 38(Barton et al, 1980) (350mg, 0.74mmol) and K₂CO₃A mixture of (514mg, 3.72mmol) in MeOH (5mL) was stirred at room temperature for 4 h. Ether was added and the mixture was washed with water. Mixed aqueous phase with CH₂Cl₂And (4) extracting. The combined organic extracts were extracted with MgSO₄Dried and concentrated. The residue obtained is dissolved in CH₂Cl₂(10 mL). NaOAc (183mg, 2.23mmol) and PCC (322mg, 1.49mmol) were added at room temperature. After stirring for 2h, additional PCC (160mg, 0.74mmol) was added and stirred for an additional 1 h. A hexane/EtOAc (1: 1, 20mL) mixture was added and stirred for 5 min. The brown slurry was filtered through a pad of silica gel and the filtrate was concentrated. The resulting residue was purified by column chromatography (silica gel, 25% to 33% EtOAc in hexanes) to give compound 39(300mg, 95%) as a white solid.¹HNMR(400MHz，CDCl₃)δ5.73(d，1H，J＝1.6Hz)，4.39(m，1H)，3.49(ddd，1H，J＝2.0，4.0，10.8Hz)，3.34(dd，1H，J＝10.8，10.8Hz)，2.17-2.60(m，7H)，2.04-2.10(m，2H)，1.40-1.85(m，12H)，1.27(s，3H)，1.14(m，1H)，1.10(d，3H，J＝7.2Hz)，0.94(s，3H)，0.79(d，3H，J＝6.8Hz)；m/z 427.2(M+1)。

Compound H0001: freshly prepared LDA solution (1.0M, 0.18mL, 0.18mmol) was added dropwise to a solution of compound 39(50mg, 0.12mmol) in THF (1mL) at-78 ℃. After stirring for 45min, TsCN (43mg, 0.24mmol) in THF (0.5mL) was added. After stirring for 30min, NH was added ₄Cl (aq) solution to stop the reaction. The reaction mixture was adjusted to pH 3 with 3N HCl (aq) and extracted with EtOAc. The combined EtOAc extracts were washed with water and MgSO₄Dried and concentrated. The resulting crude product was dissolved in benzene (1mL) and DDQ (25mg, 0.13mmol) was added. The red solution was refluxed for 20min and then cooled to room temperature. NaHCO for reaction mixture₃(aq) solution and water, then MgSO₄Dried and concentrated. The resulting residue was purified by column chromatography (silica gel, 33% EtOAc in hexanes) to give H0001(19mg, 37%):¹H NMR(400MHz，CDCl₃)δ7.96(s，1H)，5.84(d，1H，J＝2.0Hz)，4.41(m，1H)，3.50(ddd，1H，J＝2.4，4.0，11.2Hz)，3.35(d，1H，J＝10.8，11.2Hz)，2.61(m，1H)，2.48-2.52(m，2H)，2.42(dd，1H，J＝7.2，8.8Hz)，2.13-2.27(m，3H)，1.51-1.88(m，9H)，1.44(m，1H)，1.39(s，3H)，1.20(m，1H)，1.11(d，3H，J＝7.2Hz)，0.97(s，3H)，0.80(d，3H，J＝6.4Hz)；¹³C NMR(100MHz，CDCl₃)δ13.1，15.0，17.1，19.0，26.3，28.7，30.1，31.3，31.3，31.7，36.4，39.5，40.9，42.5，43.1，50.9，52.2，53.666.9，79.4，109.4，113.7，116.4，120.6，162.1，164.1，190.6，203.0；m/z 450.2(M+1)。

compound 40: to a mixture of ethylene glycol (6.4g, 104mmol) and camphorsulfonic acid (40mg, 0.17mmol) in cyclohexylamine (25mL) was added dehydroepiandrosterone (5.0g, 17.3 mmol). The suspension was heated at reflux for 20h using a Dean-Stark separator (Dean-Stark trap). After cooling, the mixture was diluted with saturated sodium bicarbonate (50mL) and extracted with EtOAc (2X 100 mL). The combined organic extracts were washed with brine, over MgSO₄Drying, concentration and vacuum drying gave compound 40(5.7g, 99%) as a white solid: m/z 333.1(M + 1).

Compound 41: to a solution of compound 40(2.60g, 7.82mmol) in toluene (50mL) was added 3-methyl-2-butanone (25mL), followed by triisopropylalumina (2.4g, 11.73 mmol). The mixture was heated at reflux for 4 h. After cooling, the mixture was diluted with MTBE (100mL) and washed with saturated potassium dihydrogen phosphate (50 mL). The aqueous layer was extracted with MTBE (2X 50 mL). The combined organic extracts were washed with water, brine, MgSO ₄Drying, concentration and vacuum drying gave a crude yellow solid. The crude product was purified by column chromatography (10% EtOAc/CH)₂Cl₂) Product 41 was obtained as a white solid (1.87g, 72%): m/z 331.0(M + 1).

Compound 42: to a solution of potassium tert-butoxide in THF (57.5mL of a 1M solution, 57.5mmol) was added tert-butanol (28 mL). Compound 41(3.80g, 11.50mmol) was added and the resulting solution was stirred for 30 minutes. Allyl bromide (2.78g, 23.0mmol) was added and the mixture was mixedThe mixture was stirred for 2 h. Adding saturated NH₄Cl (aq) (50mL), and the mixture was diluted with water (50mL) and extracted with MTBE (2 × 100 mL). The combined organic extracts were washed with brine, over MgSO₄Drying, concentration and vacuum drying gave the crude product. The crude product was purified by column chromatography (10-20% EtOAc/hexanes) to afford product 42(3.46g, 73%) as a white solid: m/z 411.1(M + 1).

Compound 43: to a solution of diisopropylamine (0.54mL, 3.81mmol) in THF (5mL) at-78 deg.C was added n-BuLi (2.21mL of a 1.6M solution in hexane, 3.53 mmol). The solution was warmed to 0 ℃ and stirred for 20min, then cooled to-78 ℃. A solution of compound 42(0.58g, 1.41mmol) in THF (2mL) was added dropwise and the solution was stirred for 30 min. A solution of 4-toluenesulfonylcyanide (0.28g, 1.55mmol) in THF (2mL) was added dropwise and the solution stirred for 30 min. Water (5mL) was added and the mixture was allowed to warm to room temperature. The aqueous phase was adjusted to pH 4 with 1N HCl (aq) and extracted with MTBE (2X 50 mL). The combined organic extracts were washed with brine, over MgSO ₄Drying, concentration and vacuum drying gave the crude product. The crude product was purified by column chromatography (25% EtOAc/hexanes) to afford compound 43(0.35g, 57%) as a white foamy solid: m/z 436.1(M + 1).

Compound 44: a solution of compound 43(0.34g, 0.78mmol) in THF (10mL) was placed in N₂And (4) under the environment. 10% Pd/C (25mg) was added, and the mixture was evacuated and washed with H₂(3x) purification. At H₂The mixture was stirred for 2h under aeration and then filtered through a fine sintered filter. The filtrate was concentrated to give compound 44 as a white foamy solid (0.34g, 100%): m/z 440.2(M + 1).

Compound 63313: to a solution of compound 44(0.34g, 0.77mmol) in benzene (8mL) was added DDQ (176mg, 0.77mmol) and the solution was heated at 80 ℃ for 3 h. After cooling, the mixture was diluted with EtOAc (50mL) and saturated NaHCO₃(aq), brine wash, MgSO₄Dried and concentrated to give the crude product as dark foam. The crude product was purified by column chromatography (15-25% EtOAc/hexanes) to afford as an off-white foamSolid, impure product 63313(95mg) dissolved in THF (4mL) and 1MHCl (1mL) and stirred for 2 h. The mixture was saturated NaHCO₃(aq) diluted and extracted with EtOAc (2X 50 mL). The combined organic extracts were washed with brine, over MgSO ₄Drying, concentration and vacuum drying gave the crude product. The crude product was purified by column chromatography (15% EtOAc/hexanes) to give compound 63313(34mg, 10%) as a white foamy solid:¹H NMR(500MHz，CDCl₃)δ7.67(s，1H)，5.59(br s，1H)，2.53(dd，J＝19and 9Hz，1H)，2.44(br d，J＝19Hz，1H)，2.20-2.08(m，1H)，2.05-1.35(m，14H)，1.30(s，3H)，1.30-1.08(m，3H)，1.05-0.90(m，1H)，0.96(s，3H)，0.89(t，J＝7Hz，3H)，0.85(t，J＝7Hz，3H)；m/z 394.1(M+1)。

compound 43: to compound 42(0.87g, 2.12mmol) in CH₂Cl₂(100mL) was added the Grubbs 2 generation catalyst (90mg, 0.11mmol) and the solution was stirred at room temperature for 2 h. Most of CH₂Cl₂Removed by rotary evaporation. The crude product was purified by column chromatography (15% EtOAc/hexanes) to afford compound 43(0.80g, 99%) as a white crystalline solid: m/z 383.0(M + 1).

Compound 44: to a solution of diisopropylamine (0.39mL, 2.73mmol) in THF (5mL) at-78 deg.C was added n-BuLi (1.58mL of a 1.6M solution in hexane, 2.53 mmol). The solution was warmed to 0 ℃ and stirred for 20min, then cooled to-78 ℃. A solution of compound 43(0.387g, 1.01mmol) in THF (15mL) and toluene (10mL) was added dropwise and the solution was stirred for 30 min. A solution of 4-toluenesulfonylcyanide (0.22g, 1.21mmol) in THF (3mL) was added dropwise and the solution stirred for 30 min. Water (5mL) was added and the mixture was allowed to warm to room temperature. The aqueous phase was adjusted to pH 4 with 1N HCl (aq) and extracted with MTBE (2X 50 mL). The combined organic extracts were washed with brine, over MgSO ₄Dried, concentrated, and dried in vacuo. The crude product was purified by column chromatography (20% EtOAc/hexanes) to give compound 44(0.14g, 34%) as a white foamy solid: m/z 408.1(M + 1).

Compound 63304: oriented foodTo a solution of compound 44(0.11g, 0.27mmol) in benzene (3mL) was added DDQ (61mg, 0.27 mmol). The solution was heated at 80 ℃ for 30 min. After cooling, the mixture was diluted with MTBE (50mL) and saturated NaHCO₃(20mL), washed with brine, MgSO₄Dried and concentrated to give the crude product as a light brown foam. The crude product was purified by column chromatography (0.5-1% EtOAc/CH)₂Cl₂) Product 63304(20mg, 18%) was obtained as a white foamy solid:¹H NMR(500MHz，CDCl₃)δ7.65(s，1H)，5.78(br s，1H)，5.68(br s，1H)，5.51(br s，1H)，4.00-3.85(m，4H)，2.88(d，J＝16Hz，1H)，2.82(d，J＝16Hz，1H)，2.71(d，J＝16Hz，1H)，2.61(d，J＝16Hz，1H)，2.22-2.15(m，1H)，2.05-1.97(m，1H)，1.90-1.40(m，9H)，1.30(s，3H)，1.33-1.22(m，2H)，0.86(s，3H)；m/z 406.1(M+1)。

compound 63311: to a solution of compound 63304(0.030g, 0.074mmol) in THF (5mL) and water (1mL) was added 1N HCl (1 mL). The solution was stirred overnight and then allowed to stand for 72 h. The mixture was saturated NaHCO₃(aq) diluted and extracted with EtOAc (2X 25 mL). The combined organic extracts were washed with brine, over MgSO₄Drying, concentration and vacuum drying gave the crude product. The crude product was purified by column chromatography (15% EtOAc/hexanes) to give product 63311(17mg, 64%) as a white solid:¹H NMR(500MHz，CDCl₃)δ7.65(s，1H)，5.80(m，1H)，5.73(m，1H)，5.53(m，1H)，2.91(d，J＝16Hz，1H)，2.84(d，J＝16Hz，1H)，2.74(br d，J＝16Hz，1H)，2.62(brd，J＝16Hz，1H)，2.50(dd，J＝19and 9Hz，1H)，2.32(dt，J＝18and 5Hz，1H)，2.13(dt，J＝19and 9Hz，1H)，2.03-1.75(m，5H)，1.73-1.53(m，2H)，1.43-1.28(m，3H)，1.33(s，3H)，0.94(s，3H)；m/z 362.0(M+1)。

compound 45: a solution of compound 43(0.79g, 2.07mmol) in THF (25mL) was placed in N ₂And (4) under the environment. 5% Pd/C (100mg) was added, and the mixture was evacuated and washed with H₂(3x) purification. Mixing the mixture with H₂Stirring for 2h under aeration condition, and filtering with fine sintered filter. The filtrate was concentrated and dried in vacuo to afford compound 45(0.78g, 98%) as a white solid: m/z 385.1(M + 1).

Compound 46: to a solution of diisopropylamine (0.77mL, 5.48mmol) in THF (10mL) at-78 deg.C was added n-BuLi (3.17mL of a 1.6M solution in hexane, 5.07 mmol). The solution was warmed to 0 ℃ and stirred for 20min, then cooled to-78 ℃. A solution of compound 45(0.78g, 2.03mmol) in THF (10mL) was added dropwise, and the solution was stirred for 30 min. A solution of 4-toluenesulfonylcyanide (0.44g, 2.43mmol) in THF (5mL) was added dropwise and the solution stirred for 30 min. Adding saturated NH₄Cl (aq) (10mL), and the mixture was warmed to room temperature. The mixture was diluted with water (10mL) and extracted with MTBE (2X 50 mL). The combined organic extracts were washed with brine, over MgSO₄Drying, concentration and vacuum drying gave the crude product. The crude product was purified by column chromatography (15-20% EtOAc/hexanes) to afford compound 46(0.46g, 55%) as a white foamy solid: m/z 410.1(M + H).

Compound 63317: a solution of compound 46(0.34g, 0.83mmol) in DMF (3mL) was cooled in an ice bath. 1, 3-dibromo-5, 5-dimethylhydantoin (0.142g, 0.50mmol) was added, and the solution was stirred at 0 ℃ for 2 h. Pyridine (0.75mL) was added and the solution was heated at 55 ℃ for 21 h. After cooling, the mixture was diluted with water (10mL) and extracted with MTBE (2X 30 mL). The combined organic extracts were washed with 0.5M HCl (aq) (2X 15mL), saturated NaHCO₃(aq) (20mL), brine, over MgSO₄Drying, concentration and vacuum drying gave the crude product. The crude product was purified by column chromatography (20% EtOAc/hexanes) to give product 63317(0.258g, 76%) as a white foamy solid:¹H NMR(500MHz，CDCl₃)δ7.63(s，1H)，5.69(br s，1H)，3.98-3.83(m，4H)，2.28-2.12(m，2H)，2.10-1.96(m，2H)，1.94-1.40(m，15H)，1.35(s，3H)，1.34-1.20(m，2H)，0.91(s，3H)；m/z 408.1(M+1)。

compound 63318: to a solution of 63317(0.23g, 0.56mmol) in THF (10mL) was added 0.5N HCl (aq) (3 mL). The solution was stirred for 50 h. The mixture was saturated NaHCO₃(aq) diluted and extracted with MTBE (2X 50 mL). The combined organic extracts were washed with brine, over MgSO₄Dried, concentrated, and dried under vacuum at 50 ℃ to give product 63318(200mg, 98%) as a white foamy solid:¹H NMR(500MHz，CDCl₃)δ7.62(s，1H)，5.73(br s，1H)，2.50(dd，J＝19and 9Hz，1H)，2.34(dt，J＝19and 5Hz，1H)，2.22-2.02(m，2H)，2.01-1.55(m，15H)，1.38(s，3H)，1.38-1.24(m，2H)，0.94(s，3H)；m/z 364.1(M+1)。

compound 63329: to a solution of compound 63318(0.069g, 0.19mmol) in toluene (1mL) and isopropanol (1mL) was added triisopropylalumina (58mg, 0.28 mmol). The mixture was stirred at 75 ℃ for 20 h. The mixture was diluted with EtOAc (50mL), and diluted with 1N HCl (aq) (20mL), saturated NaHCO ₃(aq) (20mL), brine, over MgSO₄Dried and concentrated to give the crude product. The crude product was combined with the crude product from the previous run (0.082mmol grade) and purified by column chromatography (10-20% EtOAc/CH)₂Cl₂) Product 63329(20mg, 20%) was obtained as a white foamy solid:¹H NMR(500MHz，CDCl₃)δ7.61(s，1H)，5.71(br s，1H)，3.70(br t，J＝8Hz，1H)，2.29-2.06(m，3H)，1.98-1.13(m，18H)，1.38(s，3H)，1.08-0.97(m，1H)，0.83(s，3H)；m/z 366.0(M+1)。

compound 47: a solution of acetyl chloride (0.125mL) in 125mL of methanol was treated once with cholic acid (5g, 0.012 mol). The solution was stirred at room temperature for 72h at which time the solvent was concentrated in vacuo to afford compound 47(5.0g, quantitative) as a white solid: m/z 423.2(M + 1).

Compound 48: compound 47(5.0g, 0.0118mol) was suspended in toluene (150mL) and then added in one portion to silver carbonate (6.5g) on celite (celite). The reaction mixture was refluxed for 3h and then filtered hot through a frit glass funnel. The filtrate was concentrated in vacuo to give the crude product. The crude product is taken up in CH₂Cl₂In THF (7: 3) and through a silica gel pad with CH₂Cl₂THF (7: 3) elution afforded compound 48(3.3g, 66%) as a white solid: m/z 421.1(M+1)。

Compound 49: compound 48(3.3g, 7.85mmol) in 25mL CH₂Cl₂Dilute and add diisopropylethylamine (3.3mL) and chloromethyl methyl ether (1.89g, 23.5mmol), respectively. The solution was stirred at 45 ℃ overnight (. about.14 h), cooled to room temperature, and then concentrated to give a viscous liquid. The crude product was taken up in EtOAc (50mL) and saturated KH ₂PO₄(aq) solution and brine wash. Na for organic phase₂SO₄Drying, filtration and concentration of the filtrate in vacuo afforded the crude product. The crude product was purified by column chromatography (eluting with hexane/EtOAc 7: 3) to give compound 49(2.8g, 70%) as a viscous colourless liquid which solidified on standing: m/z 385.1(M + 1).

Compound 50: compound 49(0.62g, 1.21mmol) was suspended in 9mL of ethyl formate, and 30% sodium methoxide in methanol (0.5mL) was added dropwise. The reaction mixture was stirred at room temperature for about 2h, at which point the degassing was complete. To the reaction mixture was added 0.1N HCl (aq) until the solution was pH 3. Ethyl acetate (40mL) and water (40mL) were added, the organic phase was separated and MgSO₄And (5) drying. The drying agent was filtered off and the filtrate was concentrated to give the crude product. The crude product was purified by column chromatography (eluting with hexane/EtOAc 7: 3) to give product 50(0.37g) as a white solid: m/z 551.2(M + 1).

Compound 51: compound 50(0.37g, 0.67mmol) was suspended in an ethanol/water solution (30mL/5mL), and then hydroxylamine hydrochloride (0.12g) was added in one portion. The solution was stirred at 60 ℃ overnight (. about.14 h), cooled to room temperature, and concentrated in vacuo. The product is in CH₂Cl₂(20mL) and saline (20 mL). Separating the organic phase with MgSO ₄Drying, then filtration, and concentration of the filtrate afforded the crude product. The crude product was purified by column chromatography (eluting with hexane/EtOAc 7: 3) to give compound 51 as a viscous liquid (0.15g, 40% calculated from 49): m/z 548.3(M + 1).

Compound 52: compound 51(0.15g, 0.27mmol) was diluted with 1: 1 THF/methanol (5mL) and 30% sodium methoxide in methanol (0.5mL) was added dropwise.The reaction mixture was stirred at 55 ℃ for 2 h, cooled to room temperature, and then diluted with 1: 1 EtOAc: 1N HCl (aq) (40 mL). The organic phase is separated from (Na)₂SO₄) Drying, filtering and concentrating the filtrate to dryness to obtain the crude product. The crude product was purified by column chromatography (eluting with hexane/EtOAc 7: 3) to give compound 52(0.060g, 41%) as a white solid: m/z 440.1(M + 1).

Compound 63312: compound 52(0.050g, 0.093mmol) was placed in DMF (1mL) and 1, 3-dibromo-5, 5-dimethylhydantoin (0.016g, 0.055mmol) was added. The solution was stirred at room temperature for 3 hours and then treated with 0.25mL of anhydrous pyridine. The solution was added to 55 ℃ and stirred at this temperature for 3 days. The solution was cooled to room temperature and diluted with EtOAc and 1N HCl (aq) (25 mL each). The organic phase was separated, washed with water and dried over sodium sulfate. The drying agent was filtered off and the filtrate was concentrated in vacuo to give the crude product. The crude product was purified by column chromatography (gradient elution with hexane/EtOAc from 7: 3 to 1: 1) to afford product 63312(0.040g, 80%) as a white amorphous solid: ¹H NMR(500MHz，CDCl₃) δ 7.55(s, 1H), 4.70(d, J ═ 7Hz, 1H), 4.67(d, J ═ 7Hz, 1H), 4.64(d, J ═ 7Hz, 1H), 4.56(d, J ═ 7Hz, 1H), 3.77(br s, 1H), 3.72(br s, 1H), 3.66(s, 3H), 3.48(t, J ═ 16Hz, 1H), 3.39(s, 3H), 3.30(s, 3H), 2.45-2.31(m, 2H), 2.23(m, 1H), 2.15-2.02(m, 2H), 1.92-1.53(m, 11H), 1.44-1.23(m, 2H), 1.25(s, 3H), 1.08(m, 1H), 0.92(d, 3H), 0.73 (d, 3H); m/z 438, 408, and 376.

Compound 63314: compound 63312(0.025g, 0.047mmol) was placed in 2mL CH₂Cl₂Then treated with 1mL of 2M HCl in diethyl ether. The solution was stirred at room temperature overnight and then concentrated to dryness. The residue was purified by column chromatography (eluting with a gradient of 7: 3 to 1: 1 hexane: EtOAc) to give product 63314(0.010g, 48%) as a viscous colorless liquid:¹H NMR(500MHz，CDCl₃)δ7.52(s，1H)，4.06(br s，1H)，3.95(br s，1H)，3.59(s，3H)，3.54(dd，J＝17and15Hz，1H)，2.47(dd，J＝18and 4Hz，1H)，2.26(m，1H)，2.10(m，1H)，2.03-1.53(m，14H)，1.50-1.13(m，4H)，1.27(s，3H)，1.00(d，J＝6Hz，3H)，0.76(s，3H)；m/z 444.1(M+1)。

* * * * * * * * * * * * * * * *

all methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the methods and in the steps or in the sequence of steps of the methods described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are chemically or physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such substitutions and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

Reference to the literature

The following references are expressly incorporated by reference herein to the extent that they provide exemplary procedures or other details supplementary to those set forth herein.

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Claims (6)

1. A compound of the formula:

wherein:

R₁and R₂Each is independently: c is less than or equal to 12 alkyl; or

R₁And R₂Taken together and is C.ltoreq.12 alkanediyl or C.ltoreq.12 alkenediyl; and

R₂₀and R₂₂Each is independently: hydrogen;

R₂₁is hydroxyl, oxygen, sulfydryl, sulfur or alkyl siloxy with the C less than or equal to 12;

or a pharmaceutically acceptable salt, tautomer or optical isomer thereof.

2. A compound of claim 1, R₁And R₂Are both methyl groups.

3. The compound of claim 1, further defined as:

wherein R is₂₀And R₂₂Each is independently: hydrogen;

R₂₁is hydroxyl, oxygen, sulfydryl, sulfur or alkyl siloxy with the C less than or equal to 12; or a pharmaceutically acceptable salt, tautomer or optical isomer thereof.

4. The compound of claim 3, further defined by the formula:

or a pharmaceutically acceptable salt, tautomer, or optical isomer of any of these formulae described above.

5. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt, tautomer, or optical isomer of any of these formulae described above.

6. A pharmaceutical composition comprising as active ingredient a compound according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier.