WO2014118740A1

WO2014118740A1 - Macrocyclic lactone derivatives and uses thereof

Info

Publication number: WO2014118740A1
Application number: PCT/IB2014/058693
Authority: WO
Inventors: Sivaramakrishnan Hariharan; Abhijeet Sudhir KATE; Becky Mary Thomas; Asha Kulkarni-Almeida; Jacqueline Vinodkumar TRIVEDI; Mahesh Gundaji JADHAV; Meet SHAH
Original assignee: Piramal Enterprises Limited
Priority date: 2013-02-04
Filing date: 2014-01-31
Publication date: 2014-08-07

Abstract

The present invention provides compounds represented by Formula 1: wherein, R' is as defined in the specification, in all their stereoisomeric and tautomeric forms and mixtures thereof in all ratios, and their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable polymorphs and prodrugs. The invention also relates to processes for the preparation of the compounds of Formula and pharmaceutical compositions containing them. The compounds and the pharmaceutical compositions of the present invention are useful for the treatment of inflammatory disorders and/or viral infections. The compounds and the pharmaceutical compositions of the present invention are also useful for the treatment of cancer.

Description

MACROCYCLIC LACTONE DERIVATIVES AND USES THEREOF

FIELD OF THE INVENTION

The present invention relates to macrocyclic lactone derivatives represented by the compounds of Formula 1 (as described herein), to processes for their preparation, pharmaceutical compositions comprising the same, and their use in the treatment of inflammatory disorders and/or viral infections. The compounds of the present invention are also useful for the treatment of cancer. BACKGROUND OF THE INVENTION

Proinflammatory cytokines, especially TNF-a (tumor necrosis factor-a) and interleukins (IL-Ιβ, IL-6, IL-8 and IL-17) play an important role in the inflammatory process. Both, IL-1 and TNF-a are derived from mononuclear cells and macrophages and in turn induce the expression of a variety of genes that contribute to the inflammatory process.

TNF-a has been implicated as a mediator in inflammatory bowel disease, juvenile rheumatoid arthritis, refractory rheumatoid arthritis, chronic non-rheumatoid arthritis, osteoporosis/bone resorption, septic shock, endotoxic shock, atherosclerosis, ischemia- reperfusion injury, rheumatoid arthritis, amyloidosis, multiple sclerosis, chronic recurrent uveitis, psoriatic arthritis, hepatitis C virus infection, malaria, ulcerative colitis, cachexia, plasmocytoma, sepsis, endometriosis, Behcet's disease, Wegenrer's granulomatosis, autoimmune diseases such as Crohn's disease, psoriasis or ankylosing spondylitis, immune deficiency, vasculitis, common variable immunodeficiency (CVID), inflammation, chronic graft-versus-host disease, trauma and transplant rejection, adult respiratory distress syndrome, osteoarthritis, pulmonary fibrosis, recurrent ovarian cancer, lymphoproliferative disease, coronary heart disease, refractory multiple myeloma, myeloproliferative disorder, diabetes, juvenile diabetes, meningitis, skin delayed type hypersensitivity disorders, Alzheimer's disease, systemic lupus erythematosus and allergic asthma.

Interleukin-6 (IL-6) is a pleiotropic cytokine that regulates immunological reactions involved in host defence, inflammation, haematopoiesis, and oncogenesis (Blood, 74(1), 1989, 1-10).

IL-6 is implicated as a mediator in many diseases such as diabetes, atherosclerosis, depression, Alzheimer's disease, systemic lupus erythematosus, prostate cancer and rheumatoid arthritis. The first IL-6 inhibitor tocilizumab (Innovator: Roche) has been approved for the treatment of rheumatoid arthritis and another IL-6 inhibitor namely ALD518 (Innovator: Alder Biopharmaceuticals Inc.), is under clinical trials.

Non-steroidal anti-inflammatory drugs (NSAIDs) such as aspirin, diclofenac, ibuprofen or naproxen are used to manage the pain and inflammation (swelling and redness) associated with inflammatory disorders such as rheumatoid arthritis, osteoarthritis, psoriatic arthritis or ankylosing spondylitis. However, despite the widespread use of NSAIDs, many individuals cannot tolerate the doses required to treat the inflammatory disorders over a prolonged period of time as NSAIDs are known to cause gastric erosions. Moreover, it is known that NSAIDs merely treat the symptoms of the inflammatory disorders and not the cause. When patients fail to respond to NSAIDs, other drugs such as methotrexate, gold salts, D-penicillamine and corticosteroids are used. These drugs also have significant adverse effects.

Biological agents such as monoclonal antibodies namely infliximab, etanercept and adalimumab are useful as anti-inflammatory agents, but have certain drawbacks associated with their use which may include route of administration (only parenteral), high cost, allergy induction, activation of latent tuberculosis, increased risk of cancer and congestive heart disease (NICE Review, October 2007). Hence, there is a continuing need for new drugs for the treatment of inflammatory disorders.

Further, among different diseases and disorders, viral diseases are widespread infections. Probably, the most prevalent viral infection is the common cold, which is upper respiratory tract (nose and throat) infection. Other common types of viral infections include: chickenpox, influenza, dengue (a mosquito borne viral disease), herpes simplex virus (HSV), cytomegalovirus (CMV), human immunodeficiency virus (HIV/AIDS) and human papilloma virus (HPV). It is a known fact that antibiotics do not work in viral infections. Treatment of viral infections varies depending on the specific virus. Antiviral drugs minimize the severity and length of some viral infections, for example, the drugs oseltamivir (Tamiflu^®) and zanamivir (Relenza^®) may be prescribed for some cases of influenza. Acyclovir is another anti viral drug that has gained the widest acceptance for clinical use. Acyclovir, a guanine analog, is a well-known antiviral agent which is extensively used in the treatment of infections caused by herpes simplex virus- 1 (HSV-1) and herpes simplex virus (HSV-2). Antiviral drugs can be toxic to human cells. The demand for antiviral drugs is ever growing, as viruses can develop resistance to antiviral drugs.

Cancer is a broad group of diseases which is characterized by an unregulated growth and spread of cells. This rapid and uncontrollable growth of cells may invade almost any tissue of the body. Cancer is caused due to a deregulation of the signaling pathways involved in cell cycle process i.e., cell survival, cell proliferation and cell death. Being a life threatening disease, cancer is considered to be the third leading cause of death with greater than 12 million cases worldwide. It is also expected that there will be approximately 26 million new cancer cases and 17 million cancer deaths per year by 2030 (Carcinogenesis, vol.31, no.l, 2010, pp. l00-110).

There are several therapies currently available for the treatment of cancer like surgery, immunotherapy, radiation therapy, chemotherapy, bone marrow transplantation, stem cell transplantation, hormonal therapy, antiangiogenic therapy, targeted therapy, gene therapy and others. However, current treatment options for cancer have limited effectiveness and a number of side effects are associated with them. . From among the various treatment options, chemotherapy has been used since several years and is one of the most commonly used treatments for cancer. It may be either used alone for some types of cancer or in combination with other type of treatments like radiation therapy or surgery. Most often, a combination of chemotherapeutic drugs is preferred to treat a specific type of cancer. Certain chemotherapy drugs may also be given in a specific order depending on the type of cancer it is being used to treat.

Although, chemotherapy can be quite effective in treating certain cancers, it has a disadvantage that chemotherapeutic drugs reach all parts of the body, not just the cancer cells. As a result, patients suffer severe side effects during the treatment of cancers. For instance, treatment of cancer using Cisplatin (Platinol^®) is associated with side-effects such as decrease in blood cell counts, allergic reaction including a rash and/or labored breathing and kidney damage. Another anticancer agent, gemcitabine (Gemzar^®), when administered causes side effects such as fever and flu-like symptoms, particularly during long period of treatment. Hence, there is continuing need for the compounds which could effectively treat cancer and cause fewer side effects.

PCT Published application WO2011061666 describes macrocyclic lactone oxime derivatives for the treatment of cancer.

PCT Published application WO2011061667 describes use of macrocyclic lactone oxime derivatives for the treatment of inflammatory disorders.

The present invention describes new macrocyclic lactone derivatives for the treatment of inflammatory disorders and/or viral infections. The compounds of the present invention are also useful for the treatment of cancer. SUMMARY OF THE INVENTION

In one aspect, the present invention relates to a compound of Formula 1 (as described herein below) or a stereoisomer, a tautomer, pharmaceutically acceptable salt, a solvate, a polymorph or a prodrug thereof.

According to another aspect, the present invention relates to a process for the preparation of the compound of Formula 1.

According to a further aspect, the present invention relates to use of the compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof, in the treatment of diseases or disorders mediated by one or more of the pro-inflammatory cytokines selected from abnormal TNF-a and interleukins (IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17 and IL- 18).

According to a further aspect, the present invention relates to use of the compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof, in the treatment of inflammatory disorders.

According to a further aspect, the present invention relates to use of the compound of

Formula 1 or a pharmaceutically acceptable salt or a solvate thereof,, in the treatment of viral infections.

According to a further aspect, the present invention relates to use of the compound of Formula 1 , in the treatment of cancer.

According to another aspect, the present invention relates to pharmaceutical compositions comprising a compound of Formula 1, or a pharmaceutically acceptable salt or a solvate thereof and at least one pharmaceutically acceptable excipient, a carrier or a vehicle.

According to another aspect, the present invention relates to a method for the treatment of a disease or a disorder mediated by one or more pro-inflammatory cytokines selected from TNF-a and interleukins (IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17 and IL- 18), comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof.

According to another aspect, the present invention relates to a method for the treatment of inflammatory disorder, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof.

According to another aspect, the present invention relates to a method for the treatment of viral infection, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof.

According to another aspect, the present invention relates to a method for the treatment of cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula 1.

According to another aspect, the present invention relates to a use of the compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof, in the manufacture of a medicament for the treatment of inflammatory disorders.

According to another aspect, the present invention relates to a use of the compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof, in the manufacture of a medicament for the treatment of viral infection.

According to another aspect, the present invention relates to a use of the compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof, in the manufacture of a medicament for the treatment of cancer.

One or more further aspects of the present inventions are discussed in detail herein below. These and other objectives and advantages of the present invention will be apparent to those skilled in the art from the following description.

Formula 1

or its stereoisomer or a tautomer, or a pharmaceutically acceptable salt, a solvate, a polymorph or a prodrug thereof,

wherein,

R' is -(CH₂)_n-R;

n is an integer selected from 1 to 4;

R is selected from -NR1R2_, (C6-Cio)aryl or heteroaryl; and Ri and R₂ are independently selected from hydrogen and (Ci-Ce)alkyl;

wherein, each of (Ci-C₆)alkyl, (Cg-Cio)aryl and heteroaryl is unsubstituted or substituted with one or more groups independently selected from (Ci-C₆)alkyl, halo-(Ci-C₆)alkyl, halogen, cyano, amino, hydroxy, -0-(Ci-C6)alkyl, halo-(Ci-C₆)alkoxy, carboxy, (C6-Cio)aryl and heterocyclyl.

Definitions

Unless otherwise indicated, the following definitions are set forth to illustrate and define the meaning and scope of the various terms used to describe the invention herein and the appended claims. These definitions should not be interpreted in the literal sense as they are not general definitions and are relevant only for this application.

The terms "a", "an" and "the" refers to "one or more" when used in the subject specification, including the claims. Thus, for example, reference to "a compound" may include a plurality of such compounds, or reference to "a disease" or "a disorder" includes a plurality of diseases or disorders.

It should also be noted that the term "or" is generally employed in its sense including "and/or" unless the content clearly dictates otherwise.

As used herein, the term "alkyl" or "(Ci-C₆) alkyl" whether used alone or as part of a substituent group, refers to saturated aliphatic groups, including straight or branched-chain alkyl groups. If the number of carbon atoms is not specified, "alkyl" refers to (C_j-C₆)alkyl. Accordingly, a straight-chain or branched chain alkyl has six or fewer carbon atoms in its backbone, for instance, Ci-C₆ for straight chain and C3-C₆ for branched chain. Representative examples of alkyl groups include, but are not limited to, methyl, ethyl, n- propyl, isopropyl, n-butyl, t-butyl, iso-butyl, sec -butyl, n-pentyl, isopentyl, 2-pentyl, 3-pentyl, neo-pentyl and n-hexyl.

Furthermore, unless stated otherwise, the alkyl groups can be unsubstituted or substituted. The term "substituted alkyl" refers to alkyl substituted with one or more groups independently selected from (Ci-C₆) lkyl, haloalkyl, halogen, cyano, amino, hydroxy, -O- (Ci-C₆)alkyl, haloalkoxy, carboxy, aryl, heteroaryl and heterocyclyl. Representative examples of substituted alkyl group include, but are not limited to, trifluoromethyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1-aminoethyl and benzyl.

As used herein, the term "alkoxy" refers to -0-(Ci-C₆)alkyl group, an alkyl having an oxygen atom attached thereto that is bonded via the oxygen atom. Representative examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n- butoxy, isobutoxy and tert-butoxy.

The term "aryl" or "(C6-Cio)aryl" as used herein refers to monocyclic or polycyclic hydrocarbon groups having 6 to 10 ring carbon atoms in which the carbocyclic ring(s) present have a conjugated pi electron system. Representative examples of (C6-Cio)aryl groups include, but are not limited to, phenyl and naphthyl. Aryl groups can be unsubstituted or substituted by one or more substituents independently selected from halogen, hydroxy, cyano, amino, (Ci-Ce)alkyl, haloalkyl, -0-(Ci-Ce)alkyl, haloalkoxy, carboxy, aryl, heteroaryl and heterocyclyl. When the aryl group is phenyl which is monosubstituted then the substituent can be located in the 2-, 3- or 4-position. If the phenyl carries two substituents, they can be located in 2, 3-position, 2, 4-position, 2, 5-position, 2, 6-position, 3, 4-position or 3, 5- position. Representative examples of substituted phenyl groups include, but are not limited to, 4-fluorophenyl, 4-chlorophenyl, 4-cyanophenyl, 4-trifluoromethylphenyl, 4-methyl-3- trifluoromethylphenyl, 2-methyl-5-trifluoromethylphenyl, 2-methoxy-5- trifluoromethylphenyl, 4-methoxy-3-trifluoromethylphenyl, 3-methoxy-4- trifluoromethylphenyl, 3-fluoro-4-trifluoro methylphenyl, 3-fluoro-4-trifluoromethoxyphenyl and 2,3,4,5,6-pentafluorophenyl. Any carbon atom of the aryl group with unsatisfied valency is assumed to have a hydrogen atom to satisfy the valency.

The term "heteroatom" as used herein includes nitrogen (N), oxygen (O), and sulfur (S). Any heteroatom with unsatisfied valency is assumed to have a hydrogen atom to satisfy the valency.

The term "heterocycle" or heterocyclyl" includes 4- to 10- membered saturated c monocyclic, bicyclic or tricyclic ring systems containing 4 to 10 ring atoms, of which 1 to 3 can be heteroatoms independently selected from nitrogen (N), oxygen (O) or sulfur (S) atom. The heterocycle can include ring systems that do not contain any double bonds within the rings, or unsaturated heterocyclic ring systems, which contain one or more double bonds, for example, 3 double bonds within a ring provided that the resulting mono-, bi- or tricyclic heterocyclic ring system is stable. The heterocyclyl group can, for example, have 1 or 2 oxygen atoms and/or 1 or 2 sulfur atoms and/or 1 to 3 nitrogen atoms in the ring. The nitrogen or sulfur atom of the heterocyclyl can be optionally oxidized to the corresponding N- oxide, S-oxide or S-dioxide. Aromatic heterocyclyl groups can also be referred to by the customary term "heteroaryl" for which all the definitions and explanations relating to heterocyclyl group apply. Further, the heterocycle may be fused with an aromatic aryl group such as phenyl. Representative examples of heterocyclyl groups include, but are not limited to, oxetane, azetidine, thietane, tetrahydrofuran, tetrahydrothiophene, dihydropyran, tetrahydropyran, thio-dihydropyran, thiotetrahydropyran, piperidine, piperazine, morpholine, 1,3-oxazinane, 1,3-thiazinane, 4,5,6-tetrahydropyrimidine, 2,3-dihydrofuran, dihydrothiene, dihydropyridine, tetrahydropyridine, isoxazolidine, pyrazolidine, pyrrole, pyrrolidine, pyrazole, imidazole, pyrazine, pyridazine, oxazole, triazole, isoxazole, thiazole, isothiazole, furan, thiene, pyridine, pyrimidine, benzothiazole, benzimidazole, benzooxazole, indole, isoindole, isoquinoline, quinoxaline and quinoline. Unless stated otherwise, heterocyclyl or heteroaryl can be unsubstituted or substituted with one or more substituents independently selected from (Ci-C₆)alkyl, halogen, haloalkyl, hydroxy, 0-(Ci-C₆)alkyl, haloalkoxy, (C₃- C₆)cycloalkyl, aryl, heterocyclyl, heteroaryl, amino, cyano and carboxy. The substituents may be present on either the ring carbon or the ring nitrogen atom. The substituents can be present at one or more positions provided that a stable molecule results.

The term "halogen" or "halo" refers to a fluorine (F), chlorine (CI), bromine (Br), or iodine (I) atom.

Within the context of the present application and as used herein, the term "haloalkyl" refers to radicals wherein one or more of the hydrogen atoms of the alkyl group are substituted with one or more halogens. Representative examples of haloalkyl include, but are not limited to, chloromethyl, dichloromethyl, trichloromethyl, dichloroethyl, dichloropropyl, fluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, difluorochloromethyl, dichlorofluoromethyl, difluoroethyl and difluoropropyl.

Within the context of the present application and as used herein, the term "haloalkoxy" refers to radicals wherein one or more of the hydrogen atoms of the alkoxy group are substituted with one or more halogens. Representative examples of haloalkoxy group include, but are not limited to, difluoromethoxy (OCHF₂), trifluoromethoxy (OCF₃) or trifluoroethoxy (OCH₂CF₃).

The term "amino" refers to the group "-NH₂" which may be optionally substituted by one or more substituents, for example two substituents. Representative examples of substituents include, but not limited to, (Ci-C6)alkyl and aryl groups.

The term "carboxyl "refers to the group -COOH.

It will be understood that "substitution" or "substituted with" includes the implicit proviso that such a substitution is in accordance with permitted valence state of the substituted atom and the substituent, as well as represents a stable compound, which does not readily undergo undesired transformation such as by rearrangement, cyclization, or elimination. Within the context of the present invention and as used herein, the terms "compound of Formula 1", "compounds of Formula 1" and "compounds of the present invention" are used interchangeably throughout this application, and include all the stereoisomeric and tautomeric forms and mixtures thereof in all ratios, and the pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable polymorphs and pharmaceutically acceptable prodrugs thereof. The compound(s) of Formula I can also be referred to herein as "the active compound" or "the active ingredient".

The term "subject" as used herein refers to an animal, preferably a mammal, and most preferably a human. The term "mammal" used herein refers to warm-blooded vertebrate animals of the class Mammalia, including humans, characterized by a covering of hair on the skin and, in the female, milk-producing mammary glands for nourishing the young. The term mammal includes animals such as cat, dog, rabbit, bear, fox, wolf, monkey, deer, mouse, pig, human and the like.

The term, "therapeutically effective amount" as used herein means an amount of the compound of Formula 1 or of the composition comprising compound of Formula 1, sufficient to significantly induce a positive modification in the condition to be regulated or treated, but low enough to avoid undue or severe side effects within the scope of sound medical judgment. The therapeutically effective amount of the compound of Formula 1 or the composition will vary with the particular condition being treated, the age and physical condition of the end user, the severity of the condition being treated/prevented, the duration of the treatment, the nature of concurrent therapy, the specific compound or composition employed and the particular pharmaceutically acceptable carrier utilized.

As used herein, the terms "treatment" "treat" and "therapy" refer to alleviate, slow the progression, attenuation or cure of existing disease or disorder (e.g., cancer, inflammatory disorder, viral infection). The term "treatment" also includes treating the symptoms of the disease or condition (e.g. cancer or inflammation). The term "prevent" or "prevention" as used herein, refers to delaying, slowing, inhibiting or reducing the onset of the disease like cancer, inflammatory disorder and viral infection.

The term "cancer" refers to the physiological condition in mammals that is typically characterized by unregulated growth and spread of cells. A "tumor" is described as abnormal mass of tissue resulting from abnormal growth or division of cancerous cells. The word "cancer" is sometimes used interchangeably with "tumor". The cancers that can be treated using the compounds of the present invention are as described herein. The term "a disease or a disorder mediated by pro-inflammatory cytokine, TNF-a" as used herein refers to a condition in which TNF-a has a significant role in the onset or progression of that condition. The diseases or disorders mediated by pro-inflammatory cytokine, TNF-a are as described herein.

The term "a disease or a disorder mediated by pro-inflammatory cytokines, interleukins IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17 or IL-18" as used herein refers to a condition in which one or more of the specified interleukins have a significant role in the onset or progression of that condition. The diseases or disorders mediated by one or more of the specified interleukines are as described herein.

Within the context of the present invention and as used herein, the term

"stereoisomer" is a general term used for all isomers of individual compounds (in the contest of the present invention, the compounds of Formula 1) that differ only in the orientation of their atoms in space. The term stereoisomer includes mirror image isomers (enantiomers), mixtures of mirror image isomers (racemates, racemic mixtures), geometric (cis/trans or E/Z) isomers, and isomers of compounds with more than one chiral center that are not mirror images of one another (diastereoisomers).

Within the context of the present invention and as used herein, the term "tautomer" refers to the coexistence of two (or more) compounds (the compounds of Formula 1) that differ from each other only in the position of one (or more) mobile atoms and in electron distribution, for example, keto-enol tautomers.

The term "pharmaceutically acceptable salt" includes salts of the active compound i.e. the compound of formula I, which retain the desired biological activity of the subject compound and exhibit minimal undesired toxicological effects; and are prepared with a suitable acid or a base depending on the particular substituent present in the compounds.

The term "pharmaceutically acceptable solvate" or "solvates" as used herein refers to an aggregate of a molecule (in the present invention, a compound of formula I or a pharmaceutically acceptable salt thereof) with one or more solvent molecules. Such solvents for the purpose of the invention may not interfere with the biological activity of the molecule. Preferably, the solvent used is a pharmaceutically acceptable solvent. Examples of suitable pharmaceutically acceptable solvents include, without limitation, water, ethanol and acetic acid. Most preferably, the solvent used is water and the solvates obtained are referred to as hydrates. Examples for suitable solvates are the mono- or di-hydrates or alcoholates of the compounds according to the invention. Within the context of the present invention, the term "prodrugs" includes simple prodrugs of the compounds of Formula 1, for example esters and amides.

The term "prodrug" refers to compounds that are drug precursors, which following administration, release the drug in vivo via a chemical or physiological process e.g., a prodrug on being brought to the physiological pH or through an enzyme action is converted to the desired drug form.

The compounds of the present invention can be crystallized in different forms. The term "polymorph" refers to a specific crystalline form of a compound which differs only in the arrangement and/or conformation of the molecule in the crystal lattice. Polymorphs of a compound have different physical and chemical properties.

In another embodiment, the present invention relates to a compound of Formula 1, wherein,

R' is -(CH₂)„-R;

n is 1 or 2;

R is a -NRiR₂ or (C₆-Cio)aryl; and

Ri and R₂ are independently selected from hydrogen and (Ci-Ce)alkyl;

wherein, each of (Ci-C₆)alkyl and (C₆-Cio)aryl, is unsubstituted or substituted with one or more groups independently selected from (Ci-C6)alkyl, halo-(Ci-C6)alkyl, halogen, cyano, amino, hydroxy, -0-(Ci-Ce)alkyl, halo-(Ci-C6)alkoxy, carboxy, (C6-Cio)aryl and heterocyclyl;

or a stereoisomer or a tautomer thereof, or a pharmaceutically acceptable salt, a solvate, a polymorph or a prodrug thereof.

In yet another embodiment, the present invention relates to a compound of Formula 1, wherein R' is -(CH₂)_n-R;

n is 2;

R is -NRiR₂; and

Ri and R₂ are independently selected from hydrogen and (Ci-C₆)alkyl;

wherein, (Ci-Cg)alkyl, is unsubstituted or substituted with one or more groups independently selected from (Ci-C₆)alkyl, halo-(Ci-C₆)alkyl, halogen, cyano, amino, hydroxy, -0-(Ci- C₆)alkyl, halo-(Ci-C6)alkoxy, carboxy, (C₆-Cio)aryl and heterocyclyl;

n is 2;

R is -NRiR₂; and

Ri and R₂ are independently selected from hydrogen, methyl and ethyl; wherein the methyl or ethyl is unsubstituted or substituted with one or more groups independently selected from (Ci-C₆)alkyl, halo-(Ci-C6)alkyl, halogen, cyano, amino, hydroxy, -0-(Ci-C6)alkyl, halo-(Ci- C₆)alkoxy, carboxy, (C6-Cio)aryl and heterocyclyl;

n is 2;

R is -NRiR₂; and

Ri and R₂ are independently selected from hydrogen, methyl and ethyl;

or a stereoisomer or a tautomer thereof, or a pharmaceutically acceptable salt, a solvate , a polymorph or a prodrug thereof.

In yet another embodiment, the present invention relates to a compound of Formula 1, wherein,

R' is -(CH₂)„-R;

n is i; and

R is (C₆-Cio)aryl;

wherein, (C6-Cio) ryl is unsubstituted or substituted with one or more groups independently selected from halogen, hydroxy, cyano, amino, (Ci-C6)alkyl, halo-(Ci-Ce)alkyl, -0-(Ci-

C₆)alkyl, halo-(Ci-C6)alkoxy and carboxy;

R' is -(CH₂)„-R;

n is i; and

R is phenyl;

wherein phenyl is unsubstituted or substituted with one or more groups independently selected from halogen, hydroxy, cyano, amino, (Ci-C6)alkyl, halo-(Ci-C₆)alkyl, -0-(Ci- C₆)alkyl, halo-(Ci-C6)alkoxy and carboxy; or a stereoisomer or a tautomer thereof, or a pharmaceutically acceptable salt, a solvate , a polymorph or a prodrug thereof.

R' is -(CH₂)_n-R;

n is i ; and

R is phenyl, which is substituted with one or more groups independently selected from halo- (Ci-Ce)alkyl and halogen;

Representative compounds in accordance with the present invention include:

(3Z,5E, 13E,15E)-18-((5Z,6E,10E)-5-((2-Aminoethoxy)imino)-3,9-dihydroxy-4,8-dimethyl dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyloxa cyclooctadeca-3,5,13,15-tetraen-2-one;

(3Z,5E, 13E,15E)-18-((5Z,6E,10E)-3,9-Dihydroxy-4,8-dimethyl-5-((2-(methylamino)ethoxy) imino)dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7, l l,13-tetramethyl oxacyclooctadeca-3 ,5 , 13, 15-tetraen-2-one;

(3Z,5E, 13E,15E) 8-((5Z,6E,10E)-5 (2 Dimethylamino)ethoxy)imino)-3,9-dihydroxy-4,8- dimethyldodeca-6, 10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetra methyloxacyclooctadeca-3 ,5, 13 , 15-tetraen-2-one;

(3Z,5E, 13E,15E)-9-Ethyl- 18-((5Z,6E,10E)-5-((2-(ethylamino)ethoxy)imino)-3,9-dihydroxy- 4,8-dimethyldodeca-6, 10-dien-2-yl)-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyl oxacyclooctadeca-3 ,5 , 13, 15-tetraen-2-one;

(3Z,5E, 13E,15E)-18-((5Z,6E,10E)-5-((2-(Diethylamino)ethoxy)imino)-3,9-dihydroxy-4,8- dimethyldodeca-6, 10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetra methyloxacyclooctadeca-3 ,5, 13 , 15-tetraen-2-one;

(3Z,5E, 13E,15E)-18-((5Z,6E,10E)-3,9-Dihydroxy-4,8-dimethyl-5-(((4-(trifluoromethyl) benzyl)oxy)imino)dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7, 11, 13- tetramethyloxacyclooctadeca-3 ,5,13, 15-tetraen-2-one;

(3Z,5E, 13E,15E)-9-Ethyl- 18-((5Z,6E,10E)-5-(((4-fluorobenzyl)oxy)imino)-3,9-dihydroxy- 4,8-dimethyldodeca-6, 10-dien-2-yl)-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyl oxacyclooctadeca-3 ,5 , 13, 15-tetraen-2-one;

(3Z,5E, 13E,15E)-18-((5Z,6E,10E)-3,9-Dihydroxy-4,8-dimethyl-5-(((perfluorophenyl) methoxy)imino)dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7, 11,13- tetramethyloxacyclooctadeca-3 ,5,13, 15-tetraen-2-one; or

a stereoisomer, or a tautomer, or a pharmaceutically acceptable salt thereof.

The compounds of Formula 1 of the present invention also include all stereoisomeric and tautomeric forms and mixtures thereof in all ratios and their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable polymorphs and pharmaceutically acceptable prodrugs. According to another aspect, the present invention relates to processes for the preparation of the compounds of Formula 1 or pharmaceutically acceptable salts thereof using methods well known to the persons skilled in the art. For example the compounds of Formula 1 can be prepared by the process illustrated in Schemes 1 and 2 but are not limited thereto. It will be appreciated by persons skilled in the art that within certain of the processes described herein, the order of the synthetic steps employed can be varied and will depend inter alia on factors such as the nature of functional groups present in a particular substrate and the protecting group strategy (if any) to be adopted. Clearly, such factors will also influence the choice of reagent to be used in the synthetic steps.

One or more of the reagents, reactants and intermediates used in the processes depicted in the following schemes are either commercially available or can be prepared according to standard literature procedures known in the art. For ease of reference, the starting compounds and the intermediates used for the synthesis of the compounds of the present invention are referred to by the symbols A, B, C, D and E respectively.

In the description of the process for the synthesis of the compounds of Formula 1 as provided herein, the corresponding substituent groups in the various formulae representing starting compounds and intermediates have the same meanings as that for the compound of Formula 1, unless stated otherwise.

The Schemes 1 and 2, wherein the processes for the preparation of the compounds of

Formula 1 of the present invention, are depicted below. For ease of reference, the reaction steps depicted in the Schemes 1 and 2 are referred to by using, general symbols namely a, b, c, d, e and f.

A process for the preparation of intermediates including the Intermediate D [G- (CH₂)„-ONH₂. 2HC1; wherein G is selected from -NH₂, -NHCH₃, -N(CH₃)₂, -NHCH₂CH₃, or -N(CH₂CH₃)₂ and n is 2] is depicted in Scheme 1. Scheme 1

- G (CH₂)_n— ONH₂ 2HC1

D

Step a:

Hydroxylamine hydrochloride can be reacted with benzophenone (referred to as

Compound A) in a solvent such as an aqueous alcohol, in presence of a base such as sodium hydroxide, at a temperature such as reflux temperature (Reaction a). The resulting reaction mixture can be further treated with concentrated acid such as cone. HCl to obtain the corresponding benzophenone oxime (referred to as Intermediate B).

Step b:

The compound, benzophenone oxime (the Intermediate B) obtained in step a, can be dissolved in a polar solvent such as DMF and can be reacted with a base selected from sodium hydride, sodium hydroxide or potassium hydroxide for 0.5 h to 2 h and unsubstituted or substituted chloro-ethanamine [G-(CH₂)_n-Cl; wherein G is selected from -NH₂, -NHCH₃, - N(CH₃)₂, -NHCH₂CH₃, or -N(CH₂CH₃)₂ and n is 2] can be added (Reaction b). After 3 h to 6 h, the resulting reaction mixture can be poured in an ice bath and pH can be adjusted to 2-3 by using an inorganic acid such as IN HCl. Diethyl ether extraction can be carried out in acidic condition. The pH of the aqueous layer can be adjusted to 9-10 with a base such as 10 % aqueous KOH solution and can be extracted with a solvent such as diethyl ether or ethyl acetate. Organic layer can be dried over sodium sulfate and was evaporated to dryness to obtain a compound referred to as Intermediate C.

Step c:

The intermediate C as obtained in step b, can be refluxed with concentrated acid such as cone. HCl for about 2 h to 5 h (Reaction c). The reaction mixture can be further extracted using a non-polar solvent selected from diethyl ether or ethyl acetate. Aqueous layer can be concentrated and the product can be crystallized using mixture of solvents such as methanol- water (9:1), ethanol-water (9:1) or ethanol-water (8:2) to obtain a dihydrochloride referred to as Intermediate D [G-(CH₂)_n-ONH₂. 2HC1; wherein G is selected from -NH₂, -NHCH₃, - N(CH₃)₂, -NHCH₂CH₃, or -N(CH₂CH₃)₂ and n is 2].

A process for the preparation of the compounds of Formula 1 (wherein R' is as defined in one or more of the embodiments described above) is depicted in the following Scheme 2.

Scheme 2

Microbial culture PM0224355

I a

Mixture of Concanamycin A and Concanamycin C

Alkaline hydrolysis product (E)

Compound of Formula 1

Step d:

Concanamycin crude (mixture of concanamycin A and concanamycin C, in Scheme 2) can be obtained by fermentation of a culture (PM0224355) by using the method described in the PCT Published application WO2011061666 The whole broth can be extracted using a solvent selected from ethyl acetate, chloroform or dichloromethane. Concanamycin crude can be isolated by column chromatography (d) and can be characterized by spectral comparison (The Journal of Antibiotics, 1992, Vol. 45, No. 7, 1108-1116).

Step e:

Concanamycin crude (mixture of concanamycin A and concanamycin C in Scheme 2) can be subjected to alkaline hydrolysis (Reaction e) as per the procedure described in reference (Tetrahedron Letters, 1981, Vol. 22, No. 39, 3857-60), to obtain a ketone compound, referred to herein as alkaline hydrolysis product (Intermediate E), in Scheme 2. Step f: The alkaline hydrolysis product (Intermediate E) can be reacted with the Intermediate D [of formula G-(CH₂)_n-ONH₂. 2HC1; as obtained in Scheme 1, step c, wherein n is 2, G is selected from -NH₂, -NHCH₃ or -NHCH₂CH₃] (reaction f) in the presence of a protic solvent selected from methanol or ethanol. The reaction can be carried out in the presence of an acid such as acetic acid at a temperature ranging from 15 °C to 30 °C for about 3 h-10 h to obtain the compound of Formula 1 , wherein R' is -(CH₂)_n- ; n is 2, and R is selected from -NH₂, - NHCH₃ or -NHCH2CH3.

The alkaline hydrolysis product (Intermediate E) can be reacted with the Intermediate D [of formula G-(CH₂)_n-ONH₂. 2HC1; as obtained in Scheme 1, step c, wherein n is 2, G is selected from -NH (C ^ or -NH(CFt₂CH₃)₂] (Reaction f) in the presence of a base such as pyridine and a catalytic amount of dimethylaminopyridine, at a temperature ranging from 0°C to 20 °C for about 15 h-25 h to obtain the compound of Formula 1, wherein R' is - (CH₂)„-R; n is 2, and R is selected from -N(CH₃)₂ or -N(CH₂CH₃)₂.

The alkaline hydrolysis product (Intermediate E) can also be reacted with the Intermediate D [(of formula G-(CH₂)_n-ONH₂.HCl; wherein n is 1 to 4, G is selected from unsubstituted or substituted (C₆-Cio)aryl or heteroaryl group), some of these compounds such as 0-(4-trifluoromethyl)benzyl) hydroxylamine hydrochloride, 0-(4- fluorobenzyl)hydroxylamine hydrochloride and 0-(2,3,4,5,6-pentafluorobenzyl) hydroxylamine hydrochloride are commercially available] (Reaction f) in the presence of an alcoholic solvent selected from methanol or ethanol. The reaction can be carried out in the presence of an acid such as acetic acid at a temperature ranging from 15°C to 30 °C for about 3 h-10 h to obtain the compound of Formula 1, wherein R is -(CH₂)_n-R; n is an integer from 1 to 4 and R is unsubstituted or substituted (C₆-Cio)aryl or heteroaryl.

The compounds of Formula 1 obtained by the process depicted in the above Scheme 2 can be optionally converted into their prodrugs. Additionally the compounds can be separated into individual isomers by techniques well known in the art such as column chromatography.

It will be appreciated by those skilled in the art that the compounds of the present invention can also be utilized in the form of their pharmaceutically acceptable salts or solvates thereof.

With respect to the compounds of Formula 1, the present invention also includes all stereoisomeric forms and mixtures thereof in all ratios and their pharmaceutically acceptable salts. Thus, when the compounds of Formula 1 contain one or more basic groups, i.e. groups which can be protonated, they can form an addition salt with a suitable inorganic or organic acid. Examples of suitable inorganic acids include: boric acid, perchloric acid, hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid and other inorganic acids known to the person skilled in the art. Examples of suitable organic acids include: acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, fumaric acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, ethane disulfonic acid, oxalic acid, isethionic acid, ketoglutaric acid, glycerophosphoric acid, aspartic acid, picric acid, lauric acid, palmitic acid, cholic acid, pantothenic acid, alginic acid, naphthoic acid, mandelic acid, tannic acid, camphoric acid and other organic acids known to the person skilled in the art.

The compounds of Formula 1 that contain one or more acidic group can form an addition salt with a suitable base. For example, such salts of the compounds of the present invention may include their alkali metal salts such as Li, Na, and K salts, or alkaline earth metal salts like Ca, Mg salts, or aluminium salts, or salts with ammonia or salts of organic bases such as lysine, arginine, guanidine, diethanolamine, choline and tromethamine.

The present invention furthermore includes solvates of the compounds of Formula 1 , for example hydrates with water and the solvates formed with other solvents of crystallization, such as alcohols, ethers, ethyl acetate, dioxane, dimethylformamide or a lower alkyl ketone such as acetone, or mixtures thereof.

The present invention also encompasses within its scope polymorphs of the compounds of Formula 1. Polymorphs can be obtained by heating or melting the compounds of present invention followed by gradual or fast cooling. The presence of polymorphs may be determined by techniques such as IR spectroscopy, solid probe NMR spectroscopy, differential scanning calorimetry, or powder X-ray diffraction.

The present invention also includes prodrugs of the compounds of Formula 1, such as esters and amides.

In an aspect, the compounds of Formula 1 or the pharmaceutically acceptable salts or solvates thereof, inhibit one or more pro-inflammatory cytokines selected from TNF- (Tumor necrosis factor- a) and interleukins selected from IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL- 15, IL-17 or IL-18. Accordingly, the compounds of Formula 1 are TNF-oc inhibitors and/or interleukin inhibitors.

In an embodiment, the compounds of Formula 1 or the pharmaceutically acceptable salts or solvates thereof; inhibit one or more pro-inflammatory cytokines selected from TNF- a and the interleukin IL-6.

Thus, the compounds of Formula 1 or the pharmaceutically acceptable salts or solvates thereof, find use in the treatment of diseases or disorders mediated by one or more pro-inflammatory cytokines selected from TNF-a and interleukins (IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17 and IL-18).

In one aspect, the present invention relates to a method for the treatment of a disease or a disorder mediated by one or more pro-inflammatory cytokines selected from TNF-a and interleukins (IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17 and IL-18), comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof.

In another aspect, the present invention relates to use of a compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof, for the treatment of a disease or a disorder mediated by one or more pro-inflammatory cytokines selected from TNF-a and interleukins (IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17 and IL-18).

In an embodiment, the disease or disorder mediated by one or more pro-inflammatory cytokines selected from TNF-a and interleukins (IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17 and IL-18) is selected from the group consisting of: bone resorption disease or osteoporosis; arthritis selected from rheumatoid arthritis, osteoarthritis, juvenile rheumatoid arthritis, psoriatic arthritis, osteoarthritis, refractory rheumatoid arthritis or chronic non-rheumatoid arthritis; respiratory disorders selected from adult respiratory distress syndrome, asthma, rheumatoid asthma, systemic lupus erythematosus (SLE), pulmonary fibrosis, chronic obstructive pulmonary disease or lung sarcoidosis; kidney disorders selected from renal fibrosis, polycystic kidney disease (PKD), nephritis, diabetic nephropathy or glomerular nephritis; central nervous system (CNS) disorders selected from Alzheimer's disease, Parkinson's disease or multiple sclerosis; liver disorders; inflammatory bowel disease selected from Crohn's disease or ulcerative colitis; skin disorders selected from delayed-type hypersensitivity in skin, dermatitis, urticaria, psoriasis or scleroderma; vascular disorders selected from coronary heart disease, myocardial infarction, stroke, circulatory shock, arterial sclerosis, atherosclerosis, varicose veins, vasculitis or septic shock syndrome; allergies selected from allergic rhinitis or allergic conjunctivitis; graft versus host disease; diverticulitis; Type II diabetes; diabetic retinopathy; glaucoma and ankylosing spondylitis.

In an embodiment, the disease or disorder mediated by one or more pro-inflammatory cytokines selected from TNF-a and interleukins (IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17 and IL- 18) is an inflammatory disorder.

In an embodiment, the inflammatory disorder is selected from inflammatory bowel disease, inflammation, rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, osteoarthritis, refractory rheumatoid arthritis, chronic non-rheumatoid arthritis, osteoporosis/bone resorption, Crohn's disease, septic shock, endotoxic shock, atherosclerosis, ischemia-reperfusion injury, coronary heart disease, vasculitis, amyloidosis, multiple sclerosis, sepsis, chronic recurrent uveitis, ulcerative colitis, cachexia, psoriasis, plasmocytoma, endometriosis, Behcet's disease, Wegener's granulomatosis, meningitis, autoimmune disease, immune deficiency, common variable immunodeficiency (CVID), chronic graft-versus-host disease, adult respiratory distress syndrome, pulmory fibrosis, ankylosing spondylitis, systemic lupus erythematosus, allergic asthma and skin delayed type hypersensitivity.

In another embodiment, the inflammatory disorder is selected from inflammatory bowel disease, inflammation, rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, refractory rheumatoid arthritis, chronic non-rheumatoid arthritis, osteoarthritis, osteoporosis/bone resorption, ankylosing spondylitis, Crohn's disease, atherosclerosis, ulcerative colitis and psoriasis.

In another aspect of the present invention, there is provided a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula 1 or a stereoisomer, or a tautomer, or a pharmaceutically acceptable salt or a solvate thereof, and a pharmaceutically acceptable carrier or diluent.

In an aspect of the present invention, the pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula 1 or a pharmaceutically acceptable salt or solvate thereof, is provided for use in the treatment of a disease or a disorder associated with abnormal activity of one or more cytokines selected from Tumor Necrosis Factor-alpha (TNF-a), interleukins (IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17 and IL-18).

According to another aspect, the present invention relates to use of the compound of Formula 1 or the pharmaceutically acceptable salt or solvate thereof, in the manufacture of a medicament for the treatment of an inflammatory disorder mediated by one or more cytokines selected from Tumor Necrosis Factor-alpha (TNF-cc), and interleukins (IL-Ιβ, IL- 2, IL-6, IL-8, IL-10, IL-15, IL-17 and IL-18).

According to one aspect of the invention, the compounds of Formula 1 or the pharmaceutically acceptable salt or solvate thereof, find use in the treatment of viral infection.

According to another aspect, the present invention relates a method for the treatment of a viral infection, comprising administering to a subject in need thereof a therapeutically effective amount of the compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof.

According to another aspect, the present invention relates use of the compound of Formula 1 or a pharmaceutically acceptable salt or a solvate thereof, in the treatment of a viral infection.

According to another aspect, the present invention relates to use of the compound of Formula 1 or the pharmaceutically acceptable salt or solvate thereof, in the manufacture of a medicament for the treatment of viral infection.

According to an embodiment, the viral infection can be caused by herpes viruses (CMV, HSV 1, HSV 2, EBV, varicella zoster virus [VZV], bovid herpesvirus type 1, equid herpesvirus type 1, HHV-6, human papilloma viruses (HPV types 1-55 including carcinogenic HPV), flaviviruses (including yellow fever virus, African swine fever virus and Japanese encephalitis virus), togaviruses (including Venezuelan equine encephalomyelitis virus), influenza viruses (types A-C), retroviruses (HIV-1, HIV-2, HTLV-I, HTLV-II, SIV, FeLV, FIV, MoMSV), adenoviruses (types 1-8), poxviruses (vaccinia virus), enteroviruses (poliovirus types 1-3, Coxsackie, hepatitis A virus, and ECHO virus), gastroenteritis viruses (Norwalk viruses, rotaviruses), hantaviruses (Hantaan virus), polyomavirus, papovaviruses, rhinoviruses, parainfluenza virus types 1-4, rabies virus, respiratory synctial virus (RSV), hepatitis viruses A, B, C and E, and the like.

In further embodiment, the viral infection is caused by influenza type A, type B or type C virus.

In another further embodiment, the viral infection is caused by influenza type A virus strain selected from H1N1, H2N2, H3N2, H7N7 or H5N1.

According to one aspect, the present invention relates to the compounds of Formula 1 or the pharmaceutically acceptable salts or solvates thereof, for use in the treatment of cancer. According to another aspect, the present invention relates to a method for the treatment of cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula 1.

According to another aspect, the present invention relates to a use of compounds of Formula 1, or the pharmaceutically acceptable salts or solvates thereof, in the manufacture of a medicament for the treatment of cancer.

In an embodiment, the cancer is selected from the group consisting of carcinoma, lymphoma selected from Hodgkin's lymphoma and non-Hodgkin' s lymphoma; blastoma, sarcoma, plasmacytoma, leukemia selected from acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia, and chronic lymphocytic leukemia; squamous cell cancer (e.g., epithelial squamous cell cancer), lung cancer selected from small-cell lung cancer, non-small cell lung cancer (NSCLC), adenocarcinoma of the lung and squamous carcinoma of the lung; cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer selected from gastrointestinal cancer; brain cancer, pancreatic cancer, glioblastoma, multiple myeloma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, anal carcinoma, penile carcinoma and head cancer and neck cancer.

In an embodiment, the cancer is a solid cancer or hematologic malignancy.

In further embodiment, the cancer is hematologic malignancy.

In another embodiment, the hematologic malignancy is selected from the group consisting of multiple myeloma, acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, B-cell lymphoma, T- cell lymphoma, Hodgkin's lymphoma and non-Hodgkin' s lymphoma.

In another embodiment, the cancer is a solid cancer.

In further embodiment, the solid cancer is selected from the group consisting of carcinoma, blastoma, sarcoma, plasmacytoma, squamous cell cancer (e.g., epithelial squamous cell cancer), lung cancer selected from small-cell lung cancer, non-small cell lung cancer ("NSCLC"), adenocarcinoma of the lung and squamous carcinoma of the lung; cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer selected from gastrointestinal cancer; brain cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, anal carcinoma, penile carcinoma and head cancer and neck cancer.

In an embodiment, the cancer is selected from multiple myeloma or breast cancer.

In another embodiment, the cancer is multiple myeloma.

In yet another embodiment, the cancer is breast cancer.

The pharmaceutical compositions according to the present invention are prepared in a manner known per se and familiar to one skilled in the art. Pharmaceutically acceptable inert inorganic and/or organic carriers and/or additives can be used in addition to the compound(s) of the Formula 1, and/or its pharmaceutically acceptable salts. For the production of pills, tablets, coated tablets and hard gelatin capsules it is possible to use, for example, lactose, corn starch or derivatives thereof, gum arabic, magnesia or glucose, etc. Carriers for soft gelatin capsules and suppositories are, for example, fats, waxes, natural or hardened oils, etc. Suitable carriers for the production of solutions, for example injection solutions, or of emulsions or syrups are, for example, water, physiological sodium chloride solution or alcohols, for example, ethanol, propanol or glycerol, sugar solutions, such as glucose solutions or mannitol solutions, or a mixture of the various solvents which have been mentioned.

In addition to the active ingredients namely the compound of Formula 1, and/or its pharmaceutically acceptable salts and the carrier substances, the pharmaceutical compositions can contain additives such as, for example, fillers, antioxidants, dispersants, emulsifiers, defoamers, flavors, preservatives, solubilizers or colorants. The pharmaceutical compositions of the present invention can also contain two or more compounds of the Formula 1 and/or their pharmaceutically acceptable salts and/or their prodrugs.

The pharmaceutical compositions normally contain about 1 to 99 , for example, about 5 to 70 %, or about 10 to about 30 % by weight of the compounds of Formula 1 or their pharmaceutically acceptable salts. The amount of the compound of Formula 1, and/or its pharmaceutically acceptable salts and/or its prodrugs in the pharmaceutical compositions can, for example, be from about 5 mg to 1000 mg. The dose of the compounds of this invention, which is to be administered, can cover a wide range. The dose to be administered daily is to be selected to suit the desired effect. A dosage of about 1 mg to 100 mg/kg/day of the compound of Formula 1 and salts thereof can be administered per day. If required, higher or lower daily doses can also be administered. Actual dosage levels of the compounds of Formula 1 present as active ingredients in the pharmaceutical compositions of this invention can be varied so as to obtain an amount of the active ingredient, which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without being toxic to the patient.

The selected dosage level will depend upon a variety of factors including the activity of the particular compound of the present invention employed, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and /or materials used in combination with the particular compounds employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

The pharmaceutical compositions according to the present invention can be administered orally, for example in the form of pills, tablets, coated tablets, capsules, granules or elixirs. Administration, however, can also be carried out rectally, for example in the form of suppositories, or parenterally, for example intravenously, intramuscularly or subcutaneously, in the form of injectable sterile solutions or suspensions, or topically, for example in the form of solutions or transdermal patches, or in other ways, for example in the form of aerosols or nasal sprays.

The compounds of the present invention i.e. the compounds of formula 1 can be administered alone or in combination with other therapies suitable for the disease or disorder being treated. Where separate dosage formulations are used, the compound of formula 1 and the other therapeutic agent can be administered at essentially the same time (concurrently) or at separately staggered times (sequentially). Administration in these various ways are suitable for the present invention as long as the beneficial therapeutic effect of the compound of formula 1 or a pharmaceutically acceptable salt thereof and the other therapeutic agent are realized by the patient at substantially the same time. Such beneficial effect is achieved when the target blood level concentrations of each active drug are maintained at substantially the same time.

In an embodiment, the compound of formula 1 or a pharmaceutically acceptable salt thereof; is used in combination with a further anti-inflammatory agent; for the treatment of an inflammatory disorder (as described herein). Such an anti-inflammatory agent includes, but is not limited to, ibuprofen, naproxen, methotrexate, gold salts, D-penicillamine and corticosteroids. In another embodiment, the compound of formula 1 or a pharmaceutically acceptable salt thereof; is used in combination with a further antiviral drug; for the treatment of a viral infection (as described herein). Such an antiviral drug includes, but is not limited to, oseltamivir, zanamivir or acyclovir.

In another further embodiment, the compound of formula 1 or a pharmaceutically acceptable salt thereof; is used in combination with a further anticancer agent; for the treatment of a cancer. Such an anticancer agents include, but are not limited to, estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic/cytostatic agents, prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors and other angiogenesis inhibitors, inhibitors of cell proliferation and survival signaling, agents that interfere with receptor tyrosine kinases and cancer vaccines. The compounds of the present invention can be co-administered with radiation therapy.

It is to be understood that modifications that do not substantially affect the activity of the various embodiments of this invention are included within the scope of the invention disclosed herein.

EXAMPLES

The following terms/abbreviations/chemical formulae are employed in the examples:

μΐ. : Microlitre ATCC : American Type Culture Collection mL : Millilitre KOH : Potassium hydroxide

ng : Nanogram NaHC0₃ : Sodium bicarbonate

μ : Microgram HC1 : Hydrochloric acid

mg : Milligram DMF : Dimethylformamide

g : Gram TEA : Triethylamine

nM : Nanomolar LPS : Lipopolysaccharide

μΜ : Micromolar FBS : Fetal Bovine Serum

mM : Millimolar PMA : Phorbol myristate acetate

M : Molar PBS : Phosphate Buffer Saline

N : Normality CPM : Counts per minute

min : Minutes PMS : Phenazine methosulfate

h : Hours CFA : Complete Freunds's Adjuvant μ : Micron mpk : mg per kilogram

nm : Nanometer s.c : subcutaneous administration mm : Millimeter p.o. : oral administration

Room temperature: 25±5 °C μθ : Microcurie

C0₂ : Carbon dioxide IC₅₀ : 50 % Inhibitory concentration

DMSO : Dimethyl sulfoxide CC₅₀ : 50% Cytotoxicity concentration

HPLC: High performance liquid chromatography

hPBMC : Human peripheral blood mononuclear cells

PBMC: Peripheral blood mononuclear cells

ELISA: Enzyme Linked Immuno Sorbent Assay

RPMI : Roswell Park Memorial Institute

MTS : (3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxy phenyl)-2-(4- sulfonyl)-2H- tetrazolium)

Preparation of the intermediates (i) to (v) used in the preparation of the representative compounds of Formula 1:

Step a:

Hydroxylamine hydrochloride (6 g) was added to benzophenone (10 g, Avra

Synthesis Private Limited, India) in a mixture of ethanol (95 %, 20 mL) and water (4 mL). Further, sodium hydroxide (11 g) was added in portions. The reaction mixture was refluxed for 5 min. After cooling, it was poured into 30 mL of concentrated HCl in 200 mL water. The precipitate obtained was benzophenone oxime which was filtered and dried. Yield: 90 %.

Step b:

Benzophenone oxime (1 mole) was dissolved in DMF (10 mL) and the mixture was added to sodium hydride (6 moles) in an ice-bath under stirring. The stirring was continued for 30 min and to it unsubstituted or substituted chloro-ethanamine* (1.1 moles) was added. After 3 h, the reaction mixture was poured in ice and the pH was adjusted to 2 using IN HCl. Diethyl ether extraction was carried out in acidic condition. The pH of the aqueous layer was adjusted to 10 with 10 % KOH solution and was extracted with diethyl ether. The diethyl ether layer was dried over sodium sulfate and was evaporated to dryness.

(* The specific chloro-ethanamine used in this step depends on the desired intermediate to be prepared.)

Step c:

The product as obtained in step b was refluxed with 6N HCl for 2 h at 100 °C and the reaction mixture was extracted with diethyl ether (2 x 100 mL). The aqueous layer was dried and the resulting product was crystallized using ethanohwater (9: 1). The product obtained was unbsubstituted or substituted 2-(aminooxy)-ethanamine dihydrochloride.

The intermediates (i) to (v) as described below were prepared by the above mentioned procedure involving reaction steps (a) to (c).

Intermediate (i): 2-(Aminooxy)-ethanamine dihydrochloride was obtained by reacting benzophenone with 2-chloroethanamine.

lH NMR (CD₃OD, 300 MHz): δ 4.36 (d, 2H), 3.35 (d, 2H). Intermediate (ii): 2-(Aminooxy)-N-methylethanamine dihydrochloride was obtained by reacting benzophenone with 2-chloro-N-methyl-ethanamine.

lH NMR (CD₃OD, 300 MHz): δ 4.44 (d, 2H), 3.44 (d, 2H), 2.79 (s, 3H).

Intermediate (iii): 2-(Aminooxy)-N,N-dimethylethanamine dihydrochloride was obtained by reacting benzophenone with 2-chloro-N,N-dimethylethanamine.¹H NMR (CD₃OD, 300 MHz): δ 4.49 (d, 2H), 3.60 (d, 2H), 2.99 (s, 6H).

Intermediate (iv): 2-(Aminooxy)-N-ethylethanamine dihydrochloride was obtained by reacting benzophenone and 2-chloro-N-ethylethanamine.

1H NMR (CD₃OD, 300 MHz): δ 4.41 (d, 2H), 3.43 (d, 2H), 3.17 (q, 2H), 1.37 (t, 3H).

Intermediate (v): 2-(Aminooxy)-N,N-diethylethanamine dihydrochloride was obtained by reacting benzophenone with 2-chloro-N,N-diethylethanamine.

^XH NMR (CD3OD, 300 MHz): δ 4.49 (d, 2H), 3.60 (d, 2H), 3.35 (2H), 3.30 (2H), 1.37 (t, 6H).

Example 1

(3Z,5E,13E,15E)-18-((6E,10E)-3,9-Dihydroxy-4,8-dimethyl-5-oxododeca-6,10-dien-2-yl)-9- ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyloxacyclooctadeca-3,5,13,15-tetra en-2-one

Step 1

Isolation & purification of concanamycins

The mixture of concanamycins (concanamycin A and concanamycin C) was obtained from the microbial fermentation of PM0224355, by the method as described in PCT Published Appln. WO2011061666, which is incorporated herein by reference. As briefly described, in the method described in WO2011061666, the harvested whole broth was extracted using ethyl acetate and was concentrated to obtain the crude extract. The crude extract was further processed by column chromatography (silica gel, 3-5 % methanol in chloroform) to obtain the mixture of concanamycins (concanamycin A and concanamycin C).

Step 2

Alkaline hydrolysis of the mixture of concanamycins (concanamycin A and concanamycin C) was carried out by using 0.03 M methanolic sodium hydroxide solution, to obtain the corresponding ketone intermediate, by the method as described in reference: Tetrahedron Letters, Vol 22, No.39, 3857-60, (1981).

This product i.e. the ketone intermediate is referred to as the compound of Example 1.

Example 2

(3Z,5E, 13E, 15E)- 18-((5Z,6E, 10E)-5-((2-Aminoethoxy)imino)-3 ,9-dihydroxy-4,8-dimethyl dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyloxa cyclooctadeca-3,5,13,15-tetraen-2-one

Step 1:

The compound of Example 1 (22.2 mg) was dissolved in methanol (2 mL), and 2- (aminooxy)-ethanamine dihydrochloride (56.1 mg, intermediate (i)) was added to it at 15 °C- 20 °C and stirred. Acetic acid (0.5 mL) was added to the resulting reaction mixture and stirring was continued at the same temperature. After 10 h, ethyl acetate was added and organic layer was washed with saturated NaHC0₃ solution and brine. The organic layer was evaporated to dryness to obtain a crude product (19.3 mg).

Step 2:

The crude product obtained in step 1 was purified by using preparative HPLC:

Column: Waters X-terra, Prep RP18 OBD, 19 mm x 150 mm, 5 μ;

Gradient of Mobile Phase A: Water with 0.05 % triethyl amine (TEA) and

Mobile Phase B: Acetonitrile with 0.05 % TEA;

Flow rate: 8 mL/ min; Detection: 254 nm.

Time (min) Phase A (%) Phase B (%)

0 80 20

2 80 20

10 60 40

20 60 40

30 50 50

70 50 50

80 40 60

90 40 60

100 -120 20 80 Yield: 10 mg; 1H NMR (CDC1₃, 300 MHz): δ 6.70 (d, 1H), 6.51 (dd, 1H), 6.41 (br s, 1H), 6.19 (1H), 6.17 (1H), 5.79 (d, 1H), 5.73 (1H), 5.68 (1H), 5.48 (dd, 2H), 5.35 (dd, 1H), 5.29 (dd, 1H), 4.23 (m, 2H), 3.88 (m, 1H), 3.86 (4H), 3.66 (s, 3H), 3.60 (1H), 3.27 (s, 3H), 3.24 (isomer peak), 3.09 (m, 1H), 2.78 (m, 1H), 2.76 (m, 1H), 2.42 (m, 1H), 2.14 (m,lH), 2.06 (2H), 2.00 (s, 3H), 1.83 (br s, 3H), 1.71 (br d, 3H), 1.44 (m, 1H), 1.18 (d, 3H), 1.10 (5H), 1.04 (d, 3H), 0.96 (d, 3H), 0.91 (3H) and 0.87 (3H); Mass: ESI-MS (+)ve mode [M+H]⁺ : 732.7, (-)ve mode: [M+Cl]^": 767.

Example 3

(3Z,5E, 13E, 15E) - 18-((5Z,6E, 10E)-3 ,9-Dihydroxy-4,8-dimethyl-5-((2-(methylamino)ethoxy) imino)dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l 1,13-tetramethyl oxacyclooctadeca-3 ,5 , 13,15-tetraen-2-one

The compound of Example 1 (51.5 mg) was dissolved in methanol (2 mL), and 2- (aminooxy)-N-methylethanamine dihydrochloride (61.9 mg, intermediate (ii)) was added to it with stirring at 15°C-20 °C. Acetic acid (0.5 mL) was added to the resulting reaction mixture and stirring was continued at the same temperature. After 5 h, ethyl acetate was added and the organic layer was washed with saturated NaHC0₃ solution and brine. The organic layer was evaporated to dryness to obtain a crude product (41.8 mg).

The crude product was purified by using preparative HPLC conditions as described in step 2 of Example 2. Yield: 29.2 mg; ^lH NMR (CDC1₃, 300 MHz): δ 6.66 (d, 1H), 6.57 (dd, 1H), 6.45 (br s, 1H), 6.21 (dd, 1H), 5.83 (br d, 1H), 5.73 (1H), 5.68 (1H), 5.50 (dd, 1H), 5.30 (1H), 5.25 (1H), 4.44 (m, 1H), 4.25 (m, 2H), 3.92 (1H), 3.88 (1H), 3.85 (1H), 3.65 (s, 4H), 3.57 (1H), 3.28 (s, 3H), 3.26 (isomer peak), 3.12 (m, 1H), 3.04 (1H), 2.97 (m, 2H), 2.76 (1H), 2.60 (3H), 2.53 (isomer peak), 2.33 (2H), 2.15 (1H), 2.00 (br s, 3H), 1.84 (br s, 3H), 1.73 (3H), 1.50 (br m, 1H), 1.22 (3H), 1.17 (3H), 1.08 (2H), 1.02 (3H), 0.96 (3H), 0.90 (3H) and 0.87 (3H); Mass: ESI-MS (-)ve mode [M+Cl] ⁺:781.9. Example 4

(3Z,5E, 13E, 15E)- 18-((5Z,6E, 10E)-5-((2-(Dimethylamino)ethoxy)imino)-3 ,9-dihydroxy-4,8- dimethyldodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetra methyloxacyclooctadeca-3 ,5 , 13,15-tetraen-2-one

The compound of Example 1 (301 mg) was dissolved in pyridine (1 mL), and 2- (aminooxy)-N,N-dimethylethanamine hydrochloride (474 mg, intermediate (iii)) was added to it with stirring in an ice bath. A catalytic amount of dimethylammopyridine (DMAP) was added to the resulting reaction mixture and stirring was continued for 1 h at the same temperature, and then the reaction mixture was stirred further at room temperature for 18 h. Ethyl acetate was added to the reaction mixture and organic layer was washed with saturated NaHC0₃ solution and brine. The organic layer was evaporated to dryness, to obtain crude product (256 mg).

The crude product was purified by using preparative HPLC conditions as described in step 2 of Example 2. Yield: 160 mg; ^XH NMR (CDC1₃, 300 MHz): δ 6.69 (d, 1H), 6.54 (dd, 1H), 6.45 (s, 1H), 6.17 (1H), 6.14 (m, 1H), 5.82 (br d, 1H), 5.70 (m, 1H), 5.66 (1H), 5.48 (m, 1H), 5.45 (1H), 5.28 (m, 1H), 4.18 (m, 2H), 3.88 (m,lH), 3.86 (m, 1H), 3.81 (m, 1H), 3.78 (m, 1H), 3.70 (1H), 3.65 (s, 3H), 3.27 (s, 3H), 3.23 (isomer peak), 2.89 (m, 1H), 2.76 (m, 1H), 2.62 (dd, 2H), 2.38 (m, 1H), 2.30 (s, 6H), 2.13 (m, 1H), 2.0 (s, 3H), 1.83 (br s, 3H), 1.74 (br d, 3H), 1.72 (2H), 1.50 (br s, 1H), 1.21 (6H), 1.10 (3H), 1.07 (2H), 1.04 (d, 3H), 0.98 (br d, 3H) and 0.92 (br d, 3H); Mass: ESI-MS +ve mode [M+H]⁺ :761.3, (-)ve mode: [M+Cl]^" :794.9.

Example 5

(3Z,5E,13E,15E)-9-Ethyl-18-((5Z,6E,10E)-5-((2-(ethylamino)ethoxy)imino)-3,9-dihydroxy- 4,8-dimethyldodeca-6,10-dien-2-yl)-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyl oxacyclooctadeca-3 ,5 , 13,15-tetraen-2-one

The compound of Example 1 (52.4 mg) was dissolved in methanol (2 mL), and 2- (aminooxy)-N-ethylethanamine dihydrochloride (68.4 mg, intermediate (iv)) was added to it with stirring at 15°C -20 °C. Acetic acid (0.5 mL) was added to the resulting reaction mixture and stirring was continued at the same temperature. After 5 h, ethyl acetate was added and the organic layer was washed with saturated NaHC0₃ solution and brine. The organic layer was evaporated to dryness to obtain crude product (54.3 mg).

The crude product was purified by using preparative HPLC conditions as described in step 2 of Example 2. Yield: 31.5 mg; ^lH NMR (CDC1₃, 300 MHz): δ 6.67 (d, IH), 6.56 (dd, IH), 6.47 (br s, IH), 6.20 (dd, IH), 5.83 (br d, IH), 5.73 (IH), 5.68 (IH), 5.50 (dd, IH), 5.31 (IH), 5.23 (IH), 4.30 (m, IH), 4.22 (m, 2H), 4.13 (IH), 3.90 (IH), 3.88 (IH), 3.85 (IH), 3.65 (s, 3H), 3.58 (IH), 3.27 (s, 3H), 3.25 (isomer peak), 3.03 (m, IH), 2.95 (m, IH), 2.73 (3H), 2.15 (IH), 2.06 (4H), 2.00 (br s, 3H), 1.83 (br s, 3H), 1.73 (br d, 3H), 1.52 (br m, IH), 1.23 (t, 3H), 1.20 (3H), 1.17 (3H), 1.08 (2H), 1.04 (3H), 0.98 (3H), 0.89 (3H) and 0.87 (3H); Mass: ESI-MS (-)ve mode [M+C1]⁺ : 794.8.

Example 6

(3Z.5E, 13E, 15E) - 18-((5Z,6E, 10E)-5-((2-(Diethylamino)ethoxy)imino)-3 ,9-dihydroxy-4,8- dimethyldodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetra methyloxacyclooctadeca-3 ,5 , 13,15-tetraen-2-one

The compound of Example 1 (302 mg) was dissolved in pyridine (1 mL), and 2- (aminooxy)-N,N-diethylethanamine dihydrochloride (550 mg, intermediate (v)) was added to it with stirring in an ice bath. A catalytic amount of dimethylaminopyridine (DMAP) was added to the resulting reaction mixture and stirring was continued for 1 h at the same temperature, and further, the reaction mixture was stirred at room temperature for 18 h. Ethyl acetate was added to the reaction mixture and organic layer was washed with saturated NaHC0₃ solution and brine. The organic layer was evaporated to dryness to obtain crude product (259 mg).

The crude product was purified by using preparative HPLC conditions as described in step 2 of Example 2.

Yield: 152 mg; ^!H NMR (CDC1₃, 300 MHz): δ 6.69 (d, 1H), 6.54 (dd, 1H), 6.40 (br s,

1H), 6.21 (dd, 1H), 6.18 (1H), 5.82 (br d, 1H), 5.73 (1H), 5.68 (1H), 5.50 (br dd, 1H), 5.43 (1H), 5.30 (dd, 1H), 4.19 (m, 2H), 4.10 (1H), 3.91 (1H), 3.88 (1H), 3.85 (1H), 3.65 (s, 4H), 3.26 (s, 3H), 3.23 (isomer peak), 2.90 (m, 1H), 2.80 (1H), 2.79 (m, 2H), 2.63 (m, 4H), 2.38 (1H), 2.14 (1H), 2.00 (br s, 3H), 1.82 (br s, 3H), 1.80 (2H), 1.71 (br d, 3H), 1.50 (br m, 1H), 1.27 (t, 6H), 1.18 (d, 3H), 1.09 (3H), 1.07 (2H), 1.04 (3H), 1.02 (3H), 0.96 (d, 3H) and 0.90 (br m, 3H); Mass: ESI-MS (-)ve mode [M+Cl]^": 823.

Example 7

(3Z,5E,13E,15E)-18-((5Z,6E,10E)-3,9-Dihydroxy-4,8-dimethyl-5-(((4-(trifluoromethyl) benzyl)oxy)imino)dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7, 11,13- tetramethyloxacyclooctadeca-3 ,5,13, 15-tetraen-2-one

The compound of Example 1 (56 mg) was dissolved in methanol (2 mL), and 0-(4- trifluoromethyl)benzyl)hydroxylamine hydrochloride (189 mg, Vyas Biosciences, India) was added with stirring at 15°C -20 °C. Acetic acid (0.5 mL) was added to the resulting reaction mixture and stirring was continued at the same temperature. After 4 h, ethyl acetate was added to the reaction mixture and organic layer was washed with saturated NaHC0₃ solution and brine. The organic layer was evaporated to dryness to obtain crude product (78 mg).

The crude product was purified by flash chromatography.

Column: Flash cartridge 4 g (redisep, silica gel); Mobile phase: 25 % ethyl acetate in petroleum ether;

Yield: 24.9 mg; ^XH NMR (CDC1₃, 300 MHz): δ 7.61 (dd, 2H), 7.46 (dd, 2H), 6.70 (d, 1H), 6.54 (dd, 1H), 6.45 (br s, 1H), 6.23 (m, 1H), 6.17 (1H), 5.82 (br d, 1H), 5.72 (1H), 5.65 (IH), 5.49 (m, IH), 5.41 (IH), 5.29 (dd, IH), 5.16 (s, 2H), 3.89 (m, IH), 3.80 (IH), 3.76 (IH), 3.73 (IH), 3.64 (s, 3H), 3.60 (isomer peak), 3.24 (s, 3H), 3.20 (isomer peak), 2.87 (br dd, IH), 2.76 (br dd, IH), 2.38 (m, IH), 2.19 (m, IH), 2.11 (m, IH), 2.00 (br s, 3H), 1.98 (2H), 1.83 (br s, 3H), 1.73 (br d, 3H), 1.51 (m, IH), 1.27 (br d, 3H), 1.17 (d, 3H), 1.10 (3H), 1.08 (m, 2H), 1.04 (d, 3H), 0.96 (d, 3H) and 0.90 (3H); Mass: ESI-MS (-)ve mode [M+Cl]^" :881.65.

Example 8

(3Z,5E,13E,15E)-9-Ethyl-18-((5Z,6E,10E)-5-(((4-fluorobenzyl)oxy)imino)-3,9-dihydroxy-

4,8-dimethyldodeca-6,10-dien-2-yl)-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyl oxacyclooctadeca-3 ,5 , 13,15-tetraen-2-one

The compound of Example 1 (53.1 mg) was dissolved in methanol (2 mL), and 0-(4- fluoro benzyl) hydroxylamine hydrochloride (114 mg, Vyas Biosciences, India) was added to it with stirring at 15°C -20 °C. Acetic acid (0.5 mL) was added to the resulting reaction mixture and stirring was continued at the same temperature. After 4 h, ethyl acetate was added and organic layer was washed with saturated NaHC(¾ solution and brine. The organic layer was evaporated to dryness to obtain crude product (80.3 mg).

The crude product was purified by flash chromatography.

Yield: 24.3 mg; ^XH NMR (CDC1₃, 300 MHz): δ 7.35 (dd, 2H), 7.05 (dd, 2H), 6.70 (d, IH), 6.54 (dd, IH), 6.45 (br s, IH), 6.23 (m, IH), 6.17 (IH), 5.82 (br d, IH), 5.72 (IH), 5.65 (IH), 5.49 (m, IH), 5.41 (IH), 5.29 (dd, IH), 5.06 (s, 2H), 3.89 (m, IH), 3.80 (IH), 3.77 (IH), 3.74 (IH), 3.65 (s, 3H), 3.61(isomer peak), 3.24 (s, 3H), 3.20 (isomer peak), 2.87 (br dd, IH), 2.76 (br dd, IH), 2.38 (m, IH), 2.19 (m, IH), 2.11 (m, IH), 2.00 (br s, 3H), 1.98 (IH), 1.83 (br s, 3H), 1.73 (br d, 3H), 1.70 (IH), 1.51 (br s, IH), 1.27 (br d, 3H), 1.17 (d, 3H), 1.10 (3H), 1.08 (m, 2H), 1.04 (d, 3H), 0.96 (d, 3H) and 0.90 (3H); Mass: ESI-MS (-)ve mode

Example 9

(3Z.5E, 13E, 15E) - 18-((5Z,6E, 10E)-3 ,9-Dihydroxy-4,8-dimethyl-5-(((perfluorophenyl) methoxy)imino)dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13- tetramethyloxacyclooctadeca-3 ,5,13, 15-tetraen-2-one

The compound of Example 1 (60 mg) was dissolved in methanol (2 mL), and O- (2,3,4, 5,6-pentafluorobenzyl) hydroxylamine hydrochloride (112.4 mg, Alfa Aesar, India) was added to it with stirring at 15°C -20 °C. Acetic acid (0.5 mL) was added to the resulting reaction mixture and stirring was continued at the same temperature. After 4 h, ethyl acetate was added and organic layer was washed with saturated NaHC0₃ solution and brine. The organic layer was evaporated to dryness to obtain crude product (88 mg).

The crude product was purified by flash chromatography.

Yield: 48.2 mg; ^XH NMR (CDC1₃, 300 MHz): δ 6.61 (d, IH), 6.49 (dd, IH), 6.41 (br s, IH), 6.16 (IH), 5.78 (br d, IH), 5.67 (2H), 5.46 (m, IH), 5.30 (dd, IH), 5.17 (2H), 3.87 (br s, 2H), 3.76 (m, IH), 3.65 (s, 3H), 3.60 (IH), 3.30 (2H), 3.23 (s, 3H), 3.19 (isomer peak), 2.89 (br d, IH), 2.76 (br dd, IH), 2.35 (m, IH), 2.21 (m, IH), 2.10 (m, IH), 2.00 (br s, 3H), 1.83 (br s, 3H), 1.72 (br d, 3H), 1.52 (br s, IH), 1.27 (2H), 1.27 (br s, 3H), 1.15 (d, 3H), 1.07 (d, 3H), 1.00 (d, 3H), 0.92 (5H), 0.92 (2H) and 0.90 (3H); Mass: ESI-MS -ve mode [M+Cl]^" : 904. Biological evaluation of the compounds

Example 10

Evaluation of inhibition of TNF-oc and IL-6 production by screening in LPS stimulated THP- 1 cells (Primary Screening)

IL-6 production by lipopolysaccharide in THP-1 cells was measured according to the method as described in the reference: The Journal of Immunology, 151, 5631-5638, (1993), the disclosure of which is incorporated by reference for the teaching of the experiment.

THP- 1 cells (ATCC) were cultured in RPMI 1640 culture medium (Gibco BRL, Pasley, UK) containing 100 U/mL penicillin and 100 mg/mL streptomycin, (100X solution, Sigma Chemical Co. St Louis, MO) containing 10 % fetal bovine serum (JRH). The cells were differentiated with phorbol myristate acetate (Sigma). The test compounds [compounds of Example 2 to Example 9, prepared as 20 mM stock in DMSO and diluted with DMSO to achieve the following final concentrations in the assay: 100, 10, 1, 0.1, 0.01, 0.001 and 0.0001 μΜ] or vehicle (0.5 % DMSO) were added to each well and the plate was incubated for 30 min at 37 °C. To this, LPS (Escherchia coli 0127:B8, Sigma Chemical Co., St. Louis, MO) prepared as 1 mg/mL stock in PBS) was added to achieve a final concentration of 1 μg/mL. Plates were incubated at 37 °C for 24 h, 5 % C0₂. Supernatants were harvested, and assayed for TNF-oc and IL-6 by ELISA as described by the manufacturer (BD Biosciences). The cells were simultaneously evaluated for cytotoxicity using CCK-8. Percent inhibition of cytokine release compared to the control was calculated. The 50 % inhibitory concentration (IC₅₀) values were calculated by a nonlinear regression method. Results are presented in Table 1.

Table 1

TNF-a IL-6

No. of y inhibition inhibition Cytotoxicit

No. Test Compound experiments IC50 (μΜ)

ICso (μΜ) IC50 (μΜ)

(N)

1 Compound of Example 2 N=3 2 + 0.5 0.29 + 0.08 3

2 Compound of Example 3 N=2 2.39 0.46 4

3 Compound of Example 4 N=3 2.25 + 0.3 0.06 + 0.01 10

4 Compound of Example 5 N=2 1.9 0.36 2

5 Compound of Example 6 N=3 3.5 0.9 >30

6 Compound of Example 7 N=3 10 0.08 ± 0.07 10

7 Compound of Example 8 N=3 >30 0.027 >30

8 Compound of Example 9 N=l >30 0.12 >30 Conclusion: Compounds of the present invention inhibit IL-6 production significantly in LPS stimulated THP- 1 cells. Example 11

Evaluation of anti-proliferation effect and inhibition of TNF-cc and IL- 17 production

Isolation of peripheral blood mononuclear cells

PBMC were obtained from healthy donors by centrifugation of heparinized venous blood over Ficoll/Hypaque solution (Histopaque- 1077, Sigma). Mononuclear cells in the buffy layer were collected and washed three times in sterile PBS with 2 % FBS. Cells were suspended in RPMI-1640 supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin and 10 % FBS.

Cytokine release assay

The cells (PBMCs isolated from healthy donors) were uniformly plated in 96-well tissue culture plates (Nunc 167008) at a concentration of lxlO⁵ cells/well. After plating, the test compounds [compounds of Example 4 and Example 8] at concentrations, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3 and 10 μΜ/mL dissolved in dimethylsulfoxide (DMSO) were added to the cells and incubated at 37 °C for 30 min in 5 % C0₂ atmosphere. The final concentration of DMSO was maintained at 0.5 %. The vehicle (0.5 % DMSO) was used as control. The cells were stimulated with 25 ng/niL of PMA (P1585, SIGMA) and 1 μΜ ionomycin (SIGMA, 10634). The plates were incubated in 5 % C0₂ air humidified atmosphere at 37 °C for 48 h. At the end of 48 h, the supernatant from the cells were collected and ELISA was performed to detect the level of IL-17 (R&D systems, DY317E) and TNF-a (BD Biosciences, 555212) in the supernatant. Results are presented in Table 2.

Lymphocyte proliferation assay by ³H thymidine uptake

The cells (hPBMCs isolated from healthy donors) were uniformly plated in 96-well tissue culture plates (Nunc 167008) at a concentration of lxlO⁵ cells/well. After plating, the test compounds [compounds of Example 4 and Example 8 respectively] at concentrations, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3 and 10 μΜ/mL dissolved in DMSO were added to the cells and incubated at 37 °C for 30 min in 5 % C0₂ atmosphere. The final concentration of DMSO was maintained at 0.5 %. The vehicle (0.5 % DMSO) was used as control. The cells were then stimulated with 25 ng/mL of PMA (P1585, SIGMA) and 1 μΜ ionomycin (SIGMA, 10634). The plates were incubated in 5 % C0₂ air humidified atmosphere at 37 °C for 48 h. At the end of 24 h cells were pulsed with 0.5 μθ well of 3 [H] -Thymidine (BRIT, LCT3). The cells were washed twice with PBS to remove excess radioactivity. Radioactivity in the incorporated cells was captured on lysis with Microscint-40, Perkin Elmer and measured by using a liquid scintillation counter (TopCount, Perkin Elmer). The anti-proliferative effect of the test compounds was measured using the following formula.

Control (CPM) - Treated (CPM)

% Anti-proliferation = X 100

Control (CPM)

Controls consisted of PBMC with PMA and ionomycin (Stimulated) and PBMC with RPMI (Un-stimulated). Results are presented in Table 2.

Table 2

AP (Stim)* - Antiproliferation (stimulated); AP (US)** - Antiproliferation (un-stimulated); Conclusion: The compounds block IL-17 and TNF-a activity in hPBMCs stimulated with PMA and ionomycin.

Example 12

Determination of anti-arthritic activity of compound of Example 4 in Collagen Induced Arthritis (CIA) model

In bred, 8 to 10 weeks old male DBA/1J mice were used for the study. Mice were maintained in the animal facility of Piramal Enterprises Limited, Mumbai, India, in accordance with the Guide for Care and Use of Animals in Scientific Research, INSA, New Delhi, India.

For induction of disease, an emulsion of collagen II and Complete Freunds's Adjuvant (CFA) in equal volumes was prepared. Collagen was added in 10 mM acetic acid to prepare 2 mg/mL solution and was kept immediately at 2-8 °C with intermittent shaking for 24 h to dissolve completely. Animals were subjected to an intradermal injection of the emulsion at the base of the tail on day 0. On day 17, animals were randomized in different groups on the basis of body weight. Average body weight of the group was in the range of 23-24 g. Mice were randomized into the various study groups, (each group having 7-8 animals), and were administered the vehicle (10 mL/kg, (p.o.) and (s.c), twice daily, Compound of Example 4 (at concentrations 0.5 mpk (s.c, once daily), 1 mpk (s.c. once daily), 2.5 mpk (p.o., twice daily) and 5 mpk (p.o., twice daily), and standard (Enbrel, (Wyeth Limited, UK), 3 mg kg s.c, once daily). Animals were scored daily for signs of the disease (articular index and paw thickness) from day 17 to the end of study (day 42).

On day 21, booster immunization with emulsion of collagen II and CFA was carried out in similar way as it was carried out on day 0. Animals were treated upto day 42. On the last day of the study (day 42), animals were euthanized after recording disease scores and hind limbs were preserved in 10 % neutral formalin buffer for histopathological evaluations. A group of naive mice was also maintained alongside. Naive animals are the animals which are neither immunized for induction of arthritis nor do they receive any treatment with a drug. This group is maintained to take care of the normal changes in the paw thickness with age. Results:

i) Standard compound, Enbrel, at 3 mg/kg, s.c, once daily (positive control group) showed a statistically significant effect as compared to vehicle control.

ii) Compound of Example 4, when administered orally at 2.5 mg/kg and 5 mg/kg showed trends towards protection in articular index with respect to vehicle control, however, it was not statistically significant,

iii) Compound of Example 4, when administered orally at 2.5 mg/kg showed significant protection in paw thickness with respect to vehicle control (p < 0.05).

iv) Compound of Example 4, when administered orally at 5 mg/kg showed a trend towards protection but it was not statistically significant,

v) Compound of Example 4, when administered subcutaneously at 0.5 and 1.0 mg kg showed significant protection in articular index (p < 0.01) with respect to vehicle control.

vi) Compound of Example 4, when administered subcutaneously at 0.5 mg /kg showed protection in paw thickness and was statistically significant (p < 0.01).

vii) Compound of Example 4, when administered subcutaneously at 1 mg/kg showed protection, however it was not statistically significant. Conclusion: The anti arthritic activity for Compound of Example 4 was observed in collagen induced arthritis (CIA) in therapeutic regimen (treatment started after onset of disease) as well as in semi therapeutic regimen (treatment started on day 17, before onset of disease). Trend towards protection was observed in therapeutic regimen. Statistically significant protection was observed in semi therapeutic regimen.

Example 13

Determination of antiviral activity of the representative compound of Formula 1.

Propagation of influenza A virus

Cells: Madin-Darby Canine Kidney (MDCK) cells (ATCC: CCL-34)

Virus: Influenza A virus (H1N1/A/PR/8/34) (ATCC: VR-1469)

Influenza A virus was propagated in MDCK cells. MDCK cells were grown in minimum essential medium (MEM) containing 10 % fetal bovine serum (FBS) at 37 °C, 5 % C0₂. When cell attained 80-90 % confluence, the monolayer was washed with MEM and infected with appropriate dilution of virus in MEM containing 2 μg/mL of TPCK-treated (Tosyl phenylalanyl chloromethyl ketone) trypsin. Virus was allowed to adsorb to the cells for 90 min. After 90 min, virus was removed and maintenance medium [MEM containing 1 μg/mL TPCK-treated trypsin, 0.2 % bovine serum albumin (BSA), 1 mM sodium pyruvate] was added and flask was incubated at 37 °C, 5 % C0₂ for 48 h till the complete disruption of cell monolayer. Flask was then subjected to 3 freeze thaw cycles for complete cell lysis and release of the virus into the culture medium. Cell debris was removed by centrifugation at 1000 rpm for 10 min at 4 °C. The supernatant obtained was stored as 100 μL· aliquots at -80 °C. Viral titer was determined by cytopathic effect (CPE) assay and hemagglutination (HA) assay and was expressed as tissue culture infectious dose 50 (TCID₅₀) and HA titer respectively.

Hemagglutination assay

For hemagglutination assay, 4-5 mL of chicken blood was collected in a heparin vacutainer tubes. Blood was centrifuged at 800xg for 10 min and the red blood cells (RBC) pellet was washed thrice with phosphate buffered saline (PBS). In a V-bottom or U-bottom 96-well plate, 50 μΐ. of PBS was mixed with 50 μΐ. of two-fold serial dilutions of influenza A virus. Control wells contained 100 μΕ of PBS without any virus. 50 μΕ of 0.5 % RBC in PBS was added to all the wells and the plate was incubated at room temperature for 20-30 min. All the wells were observed for hemagglutination. Viability assay

MDCK cells were seeded into flat-bottom 96-well plates at 1 x 10⁴/100 μΙ,ΛνεΙΙ in MEM containing 10 % FBS. Cells were allowed to adhere overnight at 37 °C, 5 % C0₂. Next day, sub-confluent MDCK cells monolayer was washed with serum free MEM. Test compound [compound of Example 4] concentrations: 0.005 nM to 50 μΜ] and zanamivir (concentrations: 0.39 nM to 50 nM) (standard drug) were prepared in maintenance medium and 100 μΕ was added in respective wells. In cell control and virus control wells 100 μΐ_^ of maintenance medium was added. Plate was then incubated at 37 °C, 5 % C0₂ for 1 h. After 1 h, 100 μΐ. of IOOTCID₅₀ of influenza A virus (H1N1) was added in all the wells except cell control wells. In cell control wells, 100 μΐ_^ of maintenance medium was added. Plate was then incubated at 37 °C, 5 % C0₂. After 24 h, 25 μΕ of supernatant was harvested for Neuraminidase (NA) assay as described below. After supernatant harvest, plate was further incubated at 37 °C, 5 % C0₂ for another 48 h. After 48 h, maintenance medium was removed and cells were washed with phosphate buffered saline (PBS). 100 μΕ of 1 : 10 diluted MTS/PMS reagent was added to each well and plate was incubated at 37 °C for 1-2 h and read at 490 nm. Percent cytotoxicity was calculated using the formula:

B - A

% cytotoxicity = X 100

B

wherein, A - absorbance of cells treated and infected

B - absorbance of cells untreated and uninfected

Neuraminidase assay

Cell supernatants (25 μΚ) were transferred to a black 96-well plate and 75 μΕ of 20 μΜ of fluorescent substrate, 2'-(4-methylumbelliferyl)-a-D-N-acetylneuraminic acid (MU- NANA) was added. After incubation of the plate at 37 °C for 1 h, 100 μΐ. of stop solution (0.1 M glycine, pH 10, 7-25 % ethanol) was added to each well and fluorescence was read with excitation and emission filters of 355 nm and 460 nm respectively.

Results: IC₅₀ for compound of Example 4 was 37.84 nM against Influenza A virus (H1N1/A/PR/8/34) and CC₅₀ was 13.28 μΜ.

Conclusion: Compound of Example 4 exhibited potent antiviral activity against influenza A virus (H1N1). The activity of the compound of Example 4 is comparable to that of the standard drug zanamivir. Example 14

i) Determination of anticancer activity of the compound of Formula 1 in Multiple Myeloma cell lines.

A growth inhibition assay using Cell Counting Kit-8 [(CCK-8)[Dojindo laboratories] was performed for testing the anti-cancer activity of the compounds of the present invention (test compounds). Multiple Myeloma cell lines U266B1, RPMI8226, H929, MM1.S and MMl.R (dexamethasone-resistant) obtained from ATCC (Rockville, MD, USA), were seeded in 96- well transparent tissue culture plate (199 μΙ_^ΛνεΙΙ) containing 15% FBS (Hyclone) and 1% Penicillin/Streptomycin (Sigma) and incubated overnight at 37° C in 5% C0₂. Control wells containing the cells treated with 0.5% DMSO were also incubated at 37° C in 5% C0₂. A stock solution (20mM) was prepared by dissolving the test compounds in DMSO. 1 μΐ of the diluted test compound was added to cells and the cells were incubated at 37° C in 5% C0₂ for 48 h. The assay was terminated by adding 15 μΐ_^ of CCK-8 reagent in each well and the plates were incubated for 3-5 h at 37°C in a humidified atmosphere of 5% C0₂. The optical density was measured at the wavelength of 450 nm in a microplate reader. Percent growth inhibition was calculated using the formula:

B - A

% growth inhibition = X 100

B

wherein, A - absorbance of cells treated with test compounds

B - absorbance of cells treated with DMSO control IC₅₀ values were calculated from the graph plotted for percent growth inhibition to the concentration of the test compound. Results are presented in Table 3.

Table 3

IC50 (μΜ)

No. Test Compound

U266B 1 RPMI8226 H929 MM1.S MMl.R

1 Compound of Example 3 ++ ++ ++ ++ ++

2 Compound of Example 4 ++ +++ +++ ++ ++

3 Compound of Example 5 ++ ++ ++ ++ ++

4 Compound of Example 6 ++ ++ +++ ++ ++

5 Compound of Example 7 +++ +++ +++ +++ +++ 6 Compound of Example 8 ++ +++ +++ +++ +++

7 Compound of Example 9 +++ +++ +++ +++ +++ corresponds to IC₅₀ ranging 0.001 μΜ - 0.1 μΜ;

corresponds to IC₅₀ ranging 0.11 μΜ - 1 μΜ;

corresponds to IC₅₀ which is >1 μΜ

Conclusion: Representative compounds of the present invention exhibited potent anti cancer activity in multiple myeloma cell lines. ii) Determination of anticancer activity of the compound of Formula 1 in breast cancer cell lines.

Breast cancer cell lines MCF7 (ER-positive), HCC1569 (ER-negative/HER2- positive), MDA-MB231 and BT549 (triple-negative) obtained from ATCC (RockviUe, MD, USA), were seeded in 96- well transparent tissue culture plate (199 μΕΛνεΙΙ) containing 15% FBS (GIBCO) and 1% Penicillin/Streptomycin (Sigma) and incubated overnight at 37° C in 5% C0₂. The cells were passaged at 80-90% confluency. They were trypsinised using Trypsin-EDTA (Sigma), washed once with PBS and grown in their respective supplemented medium. Control wells containing the cells treated with 0.5% DMSO were incubated at 37° C in 5% C0₂. A stock solution (20mM) was prepared by dissolving the representative compounds of present invention (test compounds) in DMSO. 1 μΐ of the diluted test compound was added to cells and the cells were incubated at 37° C in 5% C0₂ for 48 h. The assay was terminated by washing wells twice with PBS and cells in each well were incubated with 200 μΐ of 7μg/mL of PI solution, prepared in PBS. The plates were stored overnight at - 70°C. PI fluorescence was measured at the excitation wavelength of 544 nm and emission wavelength of 620 nm in a microplate fluorescence reader. The percent growth inhibition and IC₅₀ values were calculated as described above. Results are presented in Table 4.

Table 4

IC50 (μΜ)

No. Test Compound

MCF7 HCC1569 MDA-MB231 BT549

1 Compound of Example 2 +++ +++ +++ +++

2 Compound of Example 3 ++ ++ +++ ++

3 Compound of Example 4 +++ +++ +++ +++ 4 Compound of Example 5 +++ +++ +++ +++

5 Compound of Example 6 ++ ++ ++ ++

6 Compound of Example 7 ++ ++ ++ ++

7 Compound of Example 8 ++ ++ ++ ++

+++ corresponds to IC₅₀ ranging 0.001 μΜ - 0.1 μΜ

++ corresponds to IC₅₀ ranging 0.11 μΜ - 1 μΜ

+ corresponds to IC₅₀ which is >1 μΜ

Conclusion: Representative compounds of the present invention exhibited potent anti activity in breast cancer cell lines.

Claims

Formula 1 wherein: R' is -(CH₂)_n-R;

n is an integer selected from 1 to 4;

R is selected from -NR1R2, (C6-Cio)aryl or heteroaryl; and

Ri and R₂ are independently selected from hydrogen and (Ci-Ce)alkyl ;

wherein, each of (Ci-C6)alkyl, (C6-Cio)aryl and heteroaryl, is unsubstituted or substituted with one or more groups independently selected from (Ci-C6)alkyl, halo-(Ci-Ce)alkyl, halogen, cyano, amino, hydroxy, -0-(Ci-C₆)alkyl, halo-(Ci-C6)alkoxy, carboxy, (C6-Cio)aryl and heterocyclyl;

2. A compound according to claim 1;

wherein: R' is -(CH₂)_n-R;

n is 1 or 2 ;

R is -NRiR₂ or (C₆-C₁₀)aryl; and

Ri and R₂ are independently selected from hydrogen and (Ci-Ce)alkyl;

wherein, each of (Ci-C6)alkyl and (C6-Cio)aryl is unsubstituted or substituted with one or more groups independently selected from (Ci-C6)alkyl, halo-(Ci-C6)alkyl, halogen, cyano, amino, hydroxy, -0-(Ci-Ce)alkyl, halo-(Ci-C6)alkoxy, carboxy, (C6-Cio)aryl and heterocyclyl;

3. A compound according to claim 1 ; wherein: R' is -(CH₂)_n- ;

n is 2;

R is -NRiR₂; and

Ri and R₂ are independently selected from hydrogen and (Ci-C6)alkyl;

wherein (Ci-C₆)alkyl is unsubstituted or substituted with one or more groups independently selected from (Ci-C6)alkyl, halo-(Ci-C6)alkyl, halogen, cyano, amino, hydroxy, -0-(Ci- C₆)alkyl, halo-(Ci-C6)alkoxy, carboxy, (C6-Cio)aryl and heterocyclyl;

4. A compound according to claim 3 ;

wherein Ri and R₂ are independently selected from hydrogen, methyl and ethyl; wherein the methyl or ethyl is unsubstituted or substituted with one or more groups independently selected from (Ci-C6) lkyl, halo-(Ci-Cg)alkyl, halogen, cyano, amino, hydroxy, -0-(Ci- C₆)alkyl, halo-(Ci-C6)alkoxy, carboxy, (C6-Cio)aryl and heterocyclyl;

5. A compound according to claim 1;

wherein: R' is -(CH₂)_n-R;

n is i; and

R is (C₆-Cio)aryl;

wherein, (C6-Cio)aryl is unsustituted or substituted with one or more groups independently selected from halogen, hydroxy, cyano, amino, (Ci-C6)alkyl, halo-(Ci-C₆)alkyl, -0-(Ci- C₆)alkyl, halo-(Ci-C6)alkoxy and carboxy;

6. A compound according to claim 5;

wherein, R is phenyl which is unsustituted or substituted with one or more groups independently selected from halogen, hydroxy, cyano, amino, (Ci-Ce)alkyl, halo-(Ci- C₆)alkyl, -0-(Ci-C6)alkyl, halo-(Ci-C6)alkoxy and carboxy;

7. A compound according to claim 6;

wherein R is phenyl which is substituted with one or more groups independently selected from halo-(Ci-C6)alkyl and halogen;

8. A compound according to any one of claims 1-7 is selected from:

(3Z,5E,13E,15E)-18-((5Z,6E,10E)-5-((2-Aminoethoxy)imino)-3,9-dihydroxy-4,8-dimethyl dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyloxa cyclooctadeca-3,5,13,15-tetraen-2-one;

(3Z,5E,13E,15E)-18-((5Z,6E,10E)-3,9-Dihydroxy-4,8-dimethyl-5-((2-(methylamino)ethoxy) imino)dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyl oxacyclooctadeca-3 ,5 , 13,15-tetraen-2-one;

(3Z,5E,13E,15E)-18-((5Z,6E,10E)-5-((2-(Dimethylamino)ethoxy)imino)-3,9-dihydroxy-4,8- dimethyldodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetra methyloxacyclooctadeca-3 ,5, 13 , 15-tetraen-2-one;

(3Z,5E,13E,15E)-9-Ethyl-18-((5Z,6E,10E)-5-((2-(ethylamino)ethoxy)imino)-3,9-dihydroxy- 4,8-dimethyldodeca-6,10-dien-2-yl)-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyl oxacyclooctadeca-3 ,5 , 13,15-tetraen-2-one;

(3Z,5E,13E,15E)-18-((5Z,6E,10E)-5-((2-(Diethylamino)ethoxy)imino)-3,9-dihydroxy-4,8- dimethyldodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetra methyloxacyclooctadeca-3 ,5, 13 , 15-tetraen-2-one;

(3Z,5E,13E,15E)-18-((5Z,6E,10E)-3,9-Dihydroxy-4,8-dimethyl-5-(((4-(trifluoromethyl) benzyl)oxy)imino)dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7, 11,13- tetramethyloxacyclooctadeca-3 ,5,13, 15-tetraen-2-one;

(3Z,5E,13E,15E)-9-Ethyl-18-((5Z,6E,10E)-5-(((4-fluorobenzyl)oxy)imino)-3,9-dihydroxy- 4,8-dimethyldodeca-6,10-dien-2-yl)-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13-tetramethyl oxacyclooctadeca-3 ,5 , 13,15-tetraen-2-one; or

(3Z,5E,13E,15E)-18-((5Z,6E,10E)-3,9-Dihydroxy-4,8-dimethyl-5-(((perfluorophenyl) methoxy)imino)dodeca-6,10-dien-2-yl)-9-ethyl-8,10-dihydroxy-3,17-dimethoxy-5,7,l l,13- tetramethyloxacyclooctadeca-3 ,5,13, 15-tetraen-2-one; or

a stereoisomer, or a tautomer, or a pharmaceutically acceptable salt thereof.

9. A pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula 1 according to claim 1, or a stereoisomer, or a tautomer, or a pharmaceutically acceptable salt or a solvate thereof, and a pharmaceutically acceptable carrier or diluent.

10. A method for the treatment of diseases or disorders mediated by one or more proinflammatory cytokines selected from TNF-oc and interleukins selected from the group consisting of IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17 and IL-18, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula 1 as claimed in claim 1, or a stereoisomer, or a tautomer, or a pharmaceutically acceptable salt or a solvate thereof.

11. The method according to claim 10, wherein the diseases or disorders mediated by one or more pro-inflammatory cytokines selected from TNF-oc, IL-Ιβ, IL-2, IL-6, IL-8, IL- 10, IL-15, IL-17 and IL-18, is selected from the group consisting of: bone resorption disease

(osteoporosis); arthritis selected from rheumatoid arthritis, osteoarthritis, juvenile rheumatoid arthritis, psoriatic arthritis, osteoarthritis, refractory rheumatoid arthritis or chronic non- rheumatoid arthritis; respiratory disorders selected from adult respiratory distress syndrome, asthma, rheumatoid asthma, systemic lupus erythematosus (SLE), pulmonary fibrosis, chronic obstructive pulmonary disease or lung sarcoidosis; kidney disorders selected from renal fibrosis, polycystic kidney disease (PKD), nephritis, diabetic nephropathy or glomerular nephritis; central nervous system (CNS) disorders selected from Alzheimer's disease, Parkinson's disease or multiple sclerosis; liver disorders; inflammatory bowel disease selected from Crohn's disease or ulcerative colitis; skin disorders selected from delayed-type hypersensitivity in skin, dermatitis, urticaria, psoriasis or scleroderma; vascular disorders selected from coronary heart disease, myocardial infarction, stroke, circulatory shock, arterial sclerosis, atherosclerosis, varicose veins, vasculitis or septic shock syndrome; allergies selected from allergic rhinitis or allergic conjunctivitis; cancer selected from glioblastoma, myeloid leukemia, acute myeloid leukemia, ovarian cancer or Kaposi's sarcoma; graft versus host disease; diverticulitis; Type II diabetes; diabetic retinopathy; glaucoma and ankylosing spondylitis.

12. The method according to claim 10, wherein the disease or disorder mediated by one or more pro-inflammatory cytokines selected from TNF-a, IL-Ιβ, IL-2, IL-6, IL-8, IL-10, IL- 15, IL-17 and IL-18 is an inflammatory disorder.

13. The method according to claim 12, wherein the inflammatory disorder is selected from inflammatory bowel disease, inflammation, rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, osteoarthritis, refractory rheumatoid arthritis, chronic non- rheumatoid arthritis, osteoporosis/bone resorption, Crohn's disease, septic shock, endotoxic shock, atherosclerosis, ischemia-reperfusion injury, coronary heart disease, vasculitis, amyloidosis, multiple sclerosis, sepsis, chronic recurrent uveitis, ulcerative colitis, cachexia, psoriasis, plasmocytoma, endometriosis, Behcet's disease, Wegener's granulomatosis, meningitis, autoimmune disease, immune deficiency, common variable immunodeficiency (CVID), chronic graft-versus-host disease, adult respiratory distress syndrome, pulmory fibrosis, ankylosing spondylitis, systemic lupus erythematosus, allergic asthma or skin delayed type hypersensitivity.

14. The method according to 13, wherein the inflammatory disorder is selected from inflammatory bowel disease, inflammation, rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, refractory rheumatoid arthritis, chronic non-rheumatoid arthritis, osteoarthritis, osteoporosis/bone resorption, ankylosing spondylitis, Crohn's disease, atherosclerosis, ulcerative colitis or psoriasis.

15. A method for the treatment of viral infection, comprising administering to a subject in need thereof a therapeutically effective amount of the compound of Formula 1 as claimed in claim 1, or a stereoisomer, or a tautomer, or a pharmaceutically acceptable salt or a solvate thereof.

16. The method according to claim 15, wherein the viral infection is caused by influenza type A, B or C virus.

17. The method according to claim 16, wherein the viral infection is caused by influenza type A virus.

18. The method according to claim 17, wherein the viral infection is caused by influenza type A virus strain selected from H1N1, H2N2, H3N2, H7N7 or H5N1.

19. A method for the treatment of cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the compound of Formula 1 as claimed in claim 1, or a stereoisomer, or a tautomer, or a pharmaceutically acceptable salt or a solvate thereof.

20. The method according to claim 19, wherein the cancer is selected from carcinoma selected from hepatic carcinoma, anal carcinoma or penile carcinoma; lymphoma selected from B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma or non-Hodgkin's lymphoma; blastoma, sarcoma, plasmacytoma, leukemia selected from acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia, or chronic lymphocytic leukemia; squamous cell cancer (lung cancer selected from small-cell lung cancer or non-small cell lung cancer (NSCLC), adenocarcinoma of the lung and squamous carcinoma of the lung; cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer; brain cancer, pancreatic cancer, glioblastoma, multiple myeloma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, head cancer and neck cancer.

21. The method according to claim 19 or 20, wherein the cancer is selected from lymphoma selected from B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma or non- Hodgkin's lymphoma; leukemia selected from acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia, or chronic lymphocytic leukemia; gastric or stomach cancer; pancreatic cancer, glioblastoma, multiple myeloma, bladder cancer, hepatoma, breast cancer, colon cancer and thyroid cancer.

22. The method according to claim 19 or 20, wherein the cancer is selected from multiple myeloma or breast cancer.