COMPOSITIONS FOR THE TREATMENT OF INFLAMMATORY DISEASES

RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.

61/767,784, filed February 21, 2013, the entire contents of which are

incorporated by reference in its entirety.

STATEMENT REGARDING FEDERALLY FUNDED RESEARCH

This invention was made with government support under National

Institutes of Health Grants No. P50-DE016191 and P01-GM095467. The government has certain rights in the invention.

FIELD OF THE INVENTION

This invention relates to compounds, methods and compositions for the treatment of inflammatory diseases and related conditions.

BACKGROUND OF THE INVENTION

Inflammation is initiated as part of the immune response to infection, injury, oxidative stress or other stimuli. Although it begins as a protective and beneficial process, when it does not end properly and timely but continues uncontrollably or in an autoimmune manner, it can lead to acute or chronic (persistent) inflammation, which is linked to the pathogenesis of a wide range of diseases affecting a variety of cells, tissues and organs. The list of acute and chronic inflammatory diseases is quite large and includes some of the most common and most challenging or difficult to treat diseases, including major unmet therapeutic needs. Some examples include: cardiovascular disease, arthritis, asthma, acute lung injury, chronic obstructive pulmonary disease, cystic fibrosis, pancreatitis, systemic lupus erythematosus, Sjogren's syndrome, thyroiditis, atherosclerosis, colitis, irritable bowel disease, celiac disease, Crohn's disease, fibromyalgia, nephritis, dermatitis, acne, periodontal disease, sepsis, stroke, Alzheimer's disease, Parkinson's disease, ophthalmic inflammation, retinopathy, retinal edema, uveitis, age-related macular degeneration, diabetes, and cancer.

The treatment of inflammatory diseases typically involves the use of compounds and compositions that inhibit the actions of pro-inflammatory molecules and pathways. Typical approaches include the use of inhibitors of pro-inflammaotry enzymes (e.g. COX-1, COX- 2); antagonists of proinflammatory cytokine receptors or pro-inflammatory chemokine receptors (e.g. CXCR2); antagonists of pro-inflammatory lipid receptors (e.g. CysLTl); agonists of glucocorticoid receptors (GR), or antibodies that target proinflammatory cytokines (e.g. TNF-alpha) or signaling or growth factors (e.g. VEGF). Among these are some of the most widely used anti- inflammatory agents, such as NSAIDS, dexamethasone and other corticosteroids, the small molecule drug singulair, and the antibodies Enbrel and Avastin.

Recent discoveries and investigations on new anti-inflammatory and pro-resolving lipid mediators derived from polyunsaturated fatty acids, including omega-3 fatty acids, suggested a new approach to ending

inflammation and promoting resolution. These include the lipoxins, derived from arachidonic acid [Petasis, N. A.; Akritopoulou-Zanze, I.; Fokin, V. V.;

Bernasconi, G.; Keledjian, R.; Yang, R.; Uddin, J.; Nagulapalli, K. C; Serhan, C. N. Prostaglandins Leukot. Essent. Fatty Acids 2005, 73, 301-321], as well as the resolvins derived from eicosapentaenoic or docosahexaenoic acid [Serhan, C. N.; Petasis, N. A. Chem. Rev. 2011, 111, 5922-5943]. Endogenous molecules of this type, as well as their synthetic structural analogs, were shown to have potent anti-inflammatory and pro-resolving properties that support their potential use for the treatment of inflammatory diseases. In particular, the D- series resolvins are formed in vivo from docosahexaenoic acid in the presence or absence of aspirin, and are able to promote the ending of the inflammatory response. They also feature a well-defined stereochemistry that distinguishes them from each other, and from other related molecules of this type.

An endogenously formed lipid mediator, called resolvin D3 was detected earlier as a biosynthetically derived metabolite of docosahexaenoic acid, but its complete structure, stereochemistry and biological actions could not be determined [Serhan, C. N.; Hong, S.; Gronert, K.; Colgan, S. P.; Devchand, P. R.; Mirick, G.; Moussignac, R.-L. J. Exp. Med. 2002, 196, 1025-1037]. The first total synthesis and structural and stereochemical characterization of resolvin D3 (RvD3) and the related aspirin-triggered resolvin D3 (AT-RvD3) were recently described [Winkler, J. W.; Uddin, J.; Serhan, C. N.; Petasis, N. A. Org. Lett. 2013, 15, 1424-1427]. The anti-inflammatory and pro-resolving biological actions of this lipid mediator were also reported [Dalli, J.; Winkler, J. W.;

Colas, R. A.; Arnardottir, H.; Cheng, C.-Y. C; Chiang, N.; Petasis, N. A.;

Serhan, C. N. Chemistry & Biology (Cell) 2013, 20, 188-201].

The present invention describes the structures and therapeutic use of a new genus of small molecules, that were conceived based on the newly defined biosynthetic structure and stereochemistry (Figure 1), synthetic strategy

(Figure 2), total synthesis (Figures 3, 4, 5), stereochemical confirmation and structure matching of synthetic and biosynthetic material (Figures 6,7), as well as the identified biological actions (Figures 8,9, 10) of resolvin D3 (RvD3) and aspirin-triggered resolvin D3 (AT-RvD3). These conpounds were shown to have potent anti-inflammatory properties (Figure 8), block human neutrophil transmigration across endothelial cells, thereby regulating the size of the inflammatory exudate and reduce potential tissue damage. They also have potent pro-resolving actions (Figure 9), being potent enhancers of the uptake of apoptotic neutrophils by macrophages as well as a stimulator of IL-10. Both compounds were shown to activate human GPR32 receptor, and increase macrophage phagocytosis and efferocytosis (Figure 10).

The present invention provides compositions and methods of use for the treatment of inflammatory diseases, including empodiments of compounds and their use as anti-inflammatory and pro-resolving molecules. BRIEF SUMMARY OF THE INVENTION

This invention provides compounds, methods and compositions for the treatment of inflammatory diseases.

One embodiment of the present invention is directed to a compound selected from a group having the general formula A:

wherein:

A is hydroxy, alkoxy, aryloxy, amino, alkylamino, dialkylamino or a salt -OM, where M is a cation selected from the group consisting of ammonium, tetra-alkyl ammonium, and the cations of sodium, potassium, magnesium and zinc;

R ^a - R ^b are independently selected from a group consisting of hydrogen, alkyl, acyl, alkoxyacyl or aminoacyl;

X and Y are independently selected from a group consisting of a carbon- carbon double bond or a carbon-carbon triple bond;

Z ^a / Z ^b are selected from a group consisting of H / OR ^c or OR ^c / H, wherein R ^c is selected from a group consisting of hydrogen, alkyl, acyl, alkoxyacyl or aminoacyl; and

W is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, halo, hydroxy, alkoxy, aryloxy, acylamino, alkoxyacyloxy, aminoacyloxy, aminoacylamino, or carboxamido.

A second embodiment of the present invention is directed to a compound selected from a group having formula B or C:

wherein:

group A is hydroxy, alkoxy, aryloxy, amino, alkylamino, dialkylamino or a salt -OM, where M is a cation selected from the group consisting of ammonium, tetra-alkyl ammonium, and the cations of sodium, potassium, magnesium and zinc;

R ^a - R ^c are independently selected from a group consisting of hydrogen, alkyl, acyl, alkoxyacyl or aminoacyl;

X and Y are independently selected from a group consisting of a carbon- carbon double bond or a carbon-carbon triple bond; and

W is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, halo, hydroxy, alkoxy, aryloxy, acylamino, alkoxyacyloxy, aminoacyloxy, aminoacylamino, or carboxamido.

A third embodiment of the present invention is directed to a compound selected from a roup having formula D, E, F, G, H, or I:

wherein:

the compound has the designated R/S chirality and Z/E geometry group A is hydroxy, alkoxy, aryloxy, amino, alkylamino, dialkylamino or a salt -OM, where M is a cation selected from the group consisting of ammonium, tetra-alkyl ammonium, and the cations of sodium, potassium, magnesium and zinc;

R ^a - R ^c are independently selected from a group consisting of hydrogen, alkyl, acyl, alkoxyacyl or aminoacyl; and

In a preferred embodiment of the present invention the compounds of formula D, E, F, G, H, or I, have the designated R/S chirality and Z/E geometry, group A being hydroxy, alkoxy, or a salt, and R ^a - R ^c being hydrogen

A fourth embodiment of the present invention is directed to a compound selected from a group having a formula J or K:

wherein:

the compound has the designated R/S chirality and Z/E geometry group A is hydroxy, alkoxy, aryloxy, amino, alkylamino, dialkylamino or a salt -OM, where M is a cation selected from the group consisting of ammonium, tetra-alkyl ammonium, and the cations of sodium, potassium, magnesium and zinc;

R ^a - R ^b are independently selected from a group consisting of hydrogen, alkyl, acyl, alkoxyacyl or aminoacyl; and

W is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, halo, hydroxy, alkoxy, aryloxy, acylamino, alkoxyacyloxy, aminoacyloxy, aminoacylamido, carboxamido.

A fifth embodiment of the present invention is directed to a compound selected from a group having formula 1 (termed resolvin D3), formula 2 (termed aspirin-triggered resolvin D3), or formula 3 or 4:

A sixth embodiment of the present invention is directed to a

pharmaceutical composition, comprising a compound having a general formula selected from A-K or from compounds of formula 1 - 4 and a pharmaceutically acceptable carrier.

A sixth embodiment embodiment of the present invention is directed to a method for the treatment of inflammatory diseases, the method comprising administering to a subject an effective amount of a compound having a general formula selected from A-K or from compounds of formula 1 - 4. Preferably, the method comprises the timely local administration of a provided compound during the time course of the inflammatory disease. The methods of the present invention may comprise systemic administration via intravenous injection or via an oral formulation. Alternatively, the methods of the present invention comprise a local administration. Local delivery may be

accomplisehed via a slow release method enabled by an implant of a

degradable polymeric material containing a provided compound or

composition. Local delivery may also be accomplished via a slow release enabled by an implanted pump device.

In a seventh embodiment of the present invention, the inflammatory disease is selected from a group consisting of: cardiovascular disease, arthritis, asthma, acute lung injury, chronic obstructive pulmonary disease, cystic fibrosis, pancreatitis, systemic lupus erythematosus, Sjogren's syndrome, thyroiditis, atherosclerosis, colitis, irritable bowel disease, celiac disease, Crohn's disease, fibromyalgia, nephritis, dermatitis, acne, periodontal disease, sepsis, stroke, Alzheimer's disease, Parkinson's disease, ophthalmic

inflammation, retinopathy, retinal edema, uveitis, age-related macular degeneration, diabetes, and cancer.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1. Biosynthesis, structure and stereochemistry of RvD3 and AT-RvD3 Ref.: Winkler, J. W.; Uddin, J.; Serhan, C. N.; Petasis, N. A. Org. Lett. 2013, 15, 1424-1427.

Figure 2. Synthetic strategy for the synthesis of RvD3

Ref.: Winkler, J. W.; Uddin, J.; Serhan, C. N.; Petasis, N. A. Org. Lett. 2013, 15, 1424-1427.

Figure 3. Stereocontrolled total synthesis of key intermediates 6, 10,

9

Ref.: Winkler, J. W.; Uddin, J.; Serhan, C. N.; Petasis, N. A. Org. Lett. 2013, 15, 1424-1427.

Figure 4. Stereocontrolled total synthesis of resolvin D3 (1) and its bis-alkyne precursor 3

Ref.: Winkler, J. W.; Uddin, J.; Serhan, C. N.; Petasis, N. A. Org. Lett. 2013, 15, 1424-1427.

Figure 5. Synthesis of AT-RvD3 (2) and its bis-alkyne precursor 4 Ref.: Winkler, J. W.; Uddin, J.; Serhan, C. N.; Petasis, N. A. Org. Lett. 2013, 15, 1424-1427.

Figure 6. Stereochemical confirmation of resolvin D3

Assignment of the Z or E stereochemistry for each C = C bond using two- dimensional NMR spectroscopy. The shown Ή-Ή gCOSY spectrum of a solution of RvD3 in CD ₃ OD [9.6 x 10 ^~3 M] was acquired using a Varian VNMRS 600 MHz NMR spectrometer at 25° C on a 5 mm Triple Resonance PFG Ή and referenced to the CD3OD and an internal standard. This spectrum depicts all of the connectivities between adjacent alkenyl hydrogens (H5-H10, H19- ¾o). The colors denote a bitmap plotting method using a rainbow palette that gives depth to the positive and negative contours. The complete identification of each H-atom using this in combination with its corresponding constants (J values) permitted the unambiguous E/Z assignment of all alkenyl hydrogens Ref.: Dalli, J.; Winkler, J. W.; Colas, R. A.; Arnardottir, H.; Cheng, C.-Y. C; Chiang, N.; Petasis, N. A.; Serhan, C. N. Chemistry & Biology (Cell) 2013, 20, 188-201 Figure 7. Endogenous RvD3 and AT-RvD3 from Resolving

Inflammatory Exudates Match Synthetic Compound

(A) Endogenous RvD3 was obtained from mice injected with zymosan

(1 mg/mouse) and exudates collected at 4 hr. These were subjected to lipid mediator metabololipidomics. Selected ion chromatograms (m/z 375-147) depict murine-resolving exudate-derived RvD3.

(B) Endogenous AT-RvD3 obtained from mice administered aspirin (500 μg) and zymosan (1 mg).

(C) Synthetic isomers 1 and 2.

(D) Coinjection of resolving exudate endogenous RvD3 with synthetic isomer 1 (inset: characteristic UV-absorption spectrum)

(E) Coinjection of resolving exudate endogenous AT-RvD3 with synthetic isomer 2 (inset: characteristic UV-absorption spectrum).

Ref.: Dalli, J.; Winkler, J. W.; Colas, R. A.; Arnardottir, H.; Cheng, C.-Y. C; Chiang, N.; Petasis, N. A.; Serhan, C. N. Chemistry & Biology (Cell) 2013, 20, 188-201

Figure 8. RvD3 Is a Potent Antineutrophil and Cytokine Regulator RvD3, AT-RvD3 (10 ng/mouse), or vehicle (saline containing 0.1% EtOH) was administered i.v. 10 min prior to i.p. injection of zymosan (1 mg/mouse).

Exudates were collected 4 hr later.

(A) Cells were enumerated and PMN identified using flow cytometry.

(B and C) IL-6 (B) and IL-10 (C) levels were measured in peritoneal exudates. Results are mean ± SEM, n = 4 mice per group. ^D p < 0.05 and ^DDQ p < 0.001 versus zymosan plus vehicle.

Dorsal skin inflammation mice were injected on day six with murine recombinant TNF-a (100 ng/mouse) following administration of RvD3, AT- RvD3 (10 ng), or vehicle (saline containing 0.1% EtOH) by intrapouch injection and at 4 hr lavages obtained.

(D) PMN by flow cytometry. (E-G) Levels of (E) MCP-1, (F) IL-6, and (G) KC. Results are mean ± SEM, n = 4 mice per group. ^D p < 0.05; ^DD p < 0.01 versus TNF-a plus vehicle.

Ref.: Dalli, J.; Winkler, J. W.; Colas, R. A.; Arnardottir, H.; Cheng, C.-Y. C; Chiang, N.; Petasis, N. A.; Serhan, C. N. Chemistry & Biology (Cell) 2013, 20, 188-201

Figure 9. RvD3 and AT-RvD3 Reduce Local Prostanoids and

Leukotrienes in Acute Inflammation

Lipid mediators in peritoneal exudates collected 4 hr after zymosan

administration were assessed using LC-MS-MS metabololipidomics following solid phase extraction.

(A) Representative multiple reaction monitoring chromato grams (MRM) of selected ion pairs for arachidonic acid-derived eicosanoids. a = 6-trans-LTB ₄ and b = 6-trans, 12-epi-LTB ₄ .

(B) Representative MS-MS spectra with diagnostic ions employed for the identification of TxB2 and PGE2

(C and D) Quantification of exudate lipid mediators following (C) RvD3 and (D) AT-RvD3 administration. Results are mean ± SEM, n = 4 mice per group. ^D p < 0.05, ^DD p < 0.01 versus zymosan plus vehicle.

Ref.: Dalli, J.; Winkler, J. W.; Colas, R. A.; Arnardottir, H.; Cheng, C.-Y. C; Chiang, N.; Petasis, N. A.; Serhan, C. N. Chemistry & Biology (Cell) 2013, 20, 188-201

Figure 10. RvD3 and AT-RvD3 Proresolving Actions

(A) Human neutrophils were labeled with CFDA and incubated with RvD3, AT-RvD3, or vehicle (DPBS containing 0.1% EtOH) for 15 min (37°C) prior to assessing their transmigration across human umbilical vein endothelial cells exposed to TNF-a (10 ng/ml). Results are representative of n = 4 distinct PMN preparations.

(B-D) Increased macrophage phagocytosis and efferocytosis. Murine peritoneal resident macrophages were incubated with (B and C) RvD3 (black square), AT- RvD3 (open circle) (0.1 pM-10 nM) or (D) 1 nM of select SPM (15 min, 37°C) followed by addition of (B) FITC-zymosan or (C and D) CFDA-labeled apoptotic PMN for 1 hr . Results are mean ± SEM of n = 4, d = 3-4. ^D p < 0.05, ^DD p < 0.01, ™p < 0.001 versus vehicle; ^# p < 0.05, RvD3 versus AT-RvD3; ⁺ p < 0.05 versus RvDl.

Ref.: Dalli, J.; Winkler, J. W.; Colas, R. A.; Arnardottir, H.; Cheng, C.-Y. C; Chiang, N.; Petasis, N. A.; Serhan, C. N. Chemistry & Biology (Cell) 2013, 20, 188-201

Figure 11. RvD3 and AT-RvD3 Activate Human GPR32

(A— D) Ligand-receptor-dependent changes in impedance with CHO cells overexpressing human GPR32. Impedance was continuously recorded with real-time monitoring across cell monolayers using an ECIS system. Results are tracings obtained from incubations of RvDl, RvD3, or AT-RvD3 (100 nM each) with CHO-GPR32 cells (A) or in the presence of anti-GPR32 Ab or nonimmune rabbit serum IgG (B-D). Results are expressed as (A) means from four separate tracings with each compound or (B-D) representative of three separate experiments. IgG, immunoglobulin G.

(E and F) Human GPR32 or mock-transfected human macrophages were incubated with RvD3 or AT-RvD3 (0.1 pM-10 nM) for 15 min, followed by addition of FITC-zymosan (60 min, 37°C). Results are expressed as mean ± SEM; n = 4 macrophage preparations. ^D p < 0.05, macrophages (ΜΦ) plus GPR32 versus ΜΦ plus mock

Ref.: Dalli, J.; Winkler, J. W.; Colas, R. A.; Arnardottir, H.; Cheng, C.-Y. C; Chiang, N.; Petasis, N. A.; Serhan, C. N. Chemistry & Biology (Cell) 2013, 20, 188-201

DETAILED DESCRIPTION OF THE INVENTION

Definitions

The term "alkyl" herein used means Ci -Cio straight or branched chain alkyl, for example, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, tert-butyl, n-pentyl, i-pentyl, neo-pentyl, tert-pentyl, and the like. The term " cycloalkyl" herein means an alkyl having a C3-C8 aliphatic ring and is exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like.

The alkyl or cycloalkyl may be optionally substituted. Substituents for optionally substituted alkyl or optionall substituted cycloalkyl are hydroxy, alkoxy (e.g., methoxy and ethoxy), halogen (e.g., fluoro, chloro, bromo, and iodo), carboxy, alkoxycarbonyl, nitro, cyano, and substituted or unsubstituted amino, phenyl, benzyloxy, and the like. These substituents are able to bind them at one or more of any possible positions of the alkyl or cycloalkyl.

The term "aryl" used means monocyclic or condensed ring aromatic hydrocarbons, preferably a phenyl.

The term "heteroaryl" herein used means an aromatic heterocyclic group, preferably a 5 to 6 membered aromatic heterocyclic group, which contains one or more hetero atoms selected from the group consisting of nitrogen, oxygen and sulfur atoms in the ring.

The aryl and heteroaryl groups may be optionally substituted on the aromatic ring. Substituents for the aromatic ring of an optionally substituted aryl" and optionally substituted heteroaryl are hydroxy, alkoxy, halogen (e.g., fluoro, chloro, bromo, and iodo), carboxy, alkoxycarbonyl (e.g.,

methoxycarbonyl and ethoxycarbonyl), nitro, cyano, alkyl, optionally substituted alky, cycloalkyl and optionally substituted alkyl. These

substituents are able to bind to it at one or more of any possible position aromatic ring.

The term "acyl" refers to a -COR group, including for example alkylcarbonyl, cycloalkylcarbonyl, arylcarbonyl, or heteroarylcarbonyls, all of which may be optionally substituted.

The compounds of the invention means compounds disclosed herein. Particular compounds of the invention are compounds described herein, and pharmaceutically acceptable salts, hydrates, enantiomers, diastereomers, or stereoisomers thereof. The compounds of the invention may be identified herein by their chemical structure and/or chemical name and/or formula number and/or assigned name. Where a compound is referred to by both a chemical structure and a chemical name or formula number or assigned name, and there is a conflict between for instance, between the chemical structure and the chemical name, the chemical structure is to be accorded more weight.

Compounds

This invention provides compounds, methods and compositions for the treatment of inflammatory diseases. In particular the present invention provides compounds selected from the group of compounds consisting of compounds A-K and 1-4, as defined herein.

One embodiment of the present invention is directed to a compound selected from a group having the general formula A:

wherein:

A is hydroxy, alkoxy, aryloxy, amino, alkylamino, dialkylamino or a salt -OM, where M is a cation selected from the group consisting of ammonium, tetra-alkyl ammonium, and the cations of sodium, potassium, magnesium and zinc;

R ^a - R ^b are independently selected from a group consisting of hydrogen, alkyl, acyl, alkoxyacyl or aminoacyl;

X and Y are independently selected from a group consisting of a carbon- carbon double bond or a carbon-carbon triple bond; Z ^a / Z ^b are selected from a group consisting of H / OR ^c or OR ^c / H, wherein R ^c is selected from a group consisting of hydrogen, alkyl, acyl, alkoxyacyl or aminoacyl; and

W is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, halo, hydroxy, alkoxy, aryloxy, acylamino, alkoxyacyloxy, aminoacyloxy, aminoacylamino, or carboxamido.

A second embodiment of the present invention is directed to a compound selected from a roup having formula B or C:

wherein:

group A is hydroxy, alkoxy, aryloxy, amino, alkylamino, dialkylamino or a salt -OM, where M is a cation selected from the group consisting of ammonium, tetra-alkyl ammonium, and the cations of sodium, potassium, magnesium and zinc;

R ^a - R ^c are independently selected from a group consisting of hydrogen, alkyl, acyl, alkoxyacyl or aminoacyl;

X and Y are independently selected from a group consisting of a carbon- carbon double bond or a carbon-carbon triple bond; and

W is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, halo, hydroxy, alkoxy, aryloxy, acylamino, alkoxyacyloxy, aminoacyloxy, aminoacylamino, or carboxamido.

A third embodiment of the present invention is directed to a compound selected from a group having formula D, E, F, G, H, or I:

wherein:

the compound has the designated R/S chirality and Z/E geometry group A is hydroxy, alkoxy, aryloxy, amino, alkylamino, dialkylamino or a salt -OM, where M is a cation selected from the group consisting of ammonium, tetra-alkyl ammonium, and the cations of sodium, potassium, magnesium and zinc;

R ^a - R ^c are independently selected from a group consisting of hydrogen, alkyl, acyl, alkoxyacyl or aminoacyl; and In a preferred embodiment of the present invention the compounds of formula D, E, F, G, H, or I, have the designated R/S chirality and Z/E geometry, group A being hydroxy, alkoxy, or a salt, and R ^a — R ^c being hydrogen

A fourth embodiment of the present invention is directed to a compound selected from a group having a formula J or K:

wherein:

the compound has the designated R/S chirality and Z/E geometry group A is hydroxy, alkoxy, aryloxy, amino, alkylamino, dialkylamino or a salt -OM, where M is a cation selected from the group consisting of ammonium, tetra-alkyl ammonium, and the cations of sodium, potassium, magnesium and zinc;

R ^a - R ^b are independently selected from a group consisting of hydrogen, alkyl, acyl, alkoxyacyl or aminoacyl; and

W is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, halo, hydroxy, alkoxy, aryloxy, acylamino, alkoxyacyloxy, aminoacyloxy, aminoacylamido, carboxamido.

A fifth embodiment of the present invention is directed to a compound selected from a group having formula 1 (termed resolvin D3), formula 2 (termed as irin-triggered resolvin D3), or formula 3 or 4:

Compositions

The compounds of the present invention can be formulated as

pharmaceutical compositions. The pharmaceutical compositions of the present invention generally comprise a compound according to the present invention dissolved or dispersed in carrier. The pharmaceutical compositions, and thus the compounds they include, can be administered to a subject in need thereof, such as a human patient in a variety of forms adapted to the chosen route of administration, i.e., orally or parenterally, by intravenous, intramuscular, or subcutaneous routes.

The compositions, and thus the compounds they include, may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be

compressed into tablets, or may be incorporated directly with the food of the patient's diet. For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. The amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.

The active compound may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

The pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (e.g., glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions. Useful dosages can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art. The amount of the compound, or an active salt or derivative thereof, required for use in treatment will vary not only with the particular salt selected but also with the route of administration, the nature of the condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician.

Another aspect of the ^' present invention is directed to a method for the treatment of inflammatory diseases, comprising the timely administration of a provided neuroprotective compound. In particular, the invention provides treatment methods within up to 6h following focal ischemia, by administering a composition comprising a provided neuroprotective compound.

Another aspect of the present invention is directed to a method for the treatment of ischemic stroke, comprising the timely administration of a provided neuroprotective compound as a complex with albumin. The method comprises administering an effective amount of the compound as a complex with albumin to a patient in need thereof. In particular, the preferred timely administration for effective treatment is within 6 hours from the initiation of an ischemic event. The method generally comprises administering an effect amount of the compound to a subject in need thereof.

Another aspect of the present invention is directed to a method of administration for the provided compositions, comprising the timely systemic administration via intravenous injection or via an oral formulation.

An "effective amount" of the composition is an amount sufficient to carry out a specifically stated purpose. An "effective amount" may be determined empirically and in a routine manners in relation to the stated purpose.

Generally, the amount should be effective to beneficially treat inflammatory diseases. Preparation of the compounds

The compounds provided herein are prepared as described herein. The present invention also provides methods for the synthesis of the provided compounds according to the general strategy outlined in Figure 2 and the detailed total synthesis summarized in Figures 3, 4 and 5.

Formulation of pharmaceutical compositions

The pharmaceutical compositions provided herein contain

therapeutically effective amounts of one or more of compounds provided herein in a pharmaceutically acceptable carrier.

The compositions contain one or more compounds provided herein. The compounds are preferably formulated into suitable pharmaceutical

preparations such as solutions, suspensions, tablets, dispersible tablets, pills, capsules, powders, sustained release formulations or elixirs, for oral

administration or in sterile solutions or suspensions for parenteral

administration, as well as transdermal patch preparation and dry powder inhalers. Typically the compounds described above are formulated into pharmaceutical compositions using techniques and procedures well known in the art (see, e.g., Ansel Introduction to Pharmaceutical Dosage Forms, Fourth Edition 1985, 126).

In the compositions, effective concentrations of one or more compounds or pharmaceutically acceptable derivatives is (are) mixed with a suitable pharmaceutical carrier or vehicle. The compounds may be derivatized as the corresponding salts, esters, enol ethers or esters, acids, bases, solvates, hydrates or prodrugs prior to formulation, as described above. The

concentrations of the compounds in the compositions are effective for delivery of an amount, upon administration, that treats, prevents, or ameliorates one or more of the symptoms of conditions including, but not limited to, undesired cell proliferation, coronary restenosis, osteoporosis and syndromes characterized by chronic inflammation, autoimmune diseases and cardiovascular diseases. Typically, the compositions are formulated for single dosage

administration. To formulate a composition, the weight fraction of compound is dissolved, suspended, dispersed or otherwise mixed in a selected vehicle at an effective concentration such that the treated condition is relieved or ameliorated. Pharmaceutical carriers or vehicles suitable for administration of the compounds provided herein include any such carriers known to those skilled in the art to be suitable for the particular mode of administration.

In addition, the compounds may be formulated as the sole

pharmaceutically active ingredient in the composition or may be combined with other active ingredients. Liposomal suspensions, including tissue- targeted liposomes, such as tumor-targeted liposomes, may also be suitable as pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art. For example, liposome

formulations may be prepared as described in U.S. Patent No. 4,522,811.

Briefly, liposomes such as multilamellar vesicles (MLV's) may be formed by drying down egg phosphatidyl choline and brain phosphatidyl serine (7:3 molar ratio) on the inside of a flask. A solution of a compound provided herein in phosphate buffered saline lacking divalent cations (PBS) is added and the flask shaken until the lipid film is dispersed. The resulting vesicles are washed to remove unencapsulated compound, pelleted by centrifugation, and then resuspended in PBS.

The active compound is included in the pharmaceutically acceptable carrier in an amount sufficient to exert a therapeutically useful effect in the absence of undesirable side effects on the patient treated. The therapeutically effective concentration may be determined empirically by testing the

compounds in in vitro and in vivo systems described herein and then

extrapolated therefrom for dosages for humans.

The concentration of active compound in the pharmaceutical

composition will depend on absorption, inactivation and excretion rates of the active compound, the physicochemical characteristics of the compound, the dosage schedule, and amount administered as well as other factors known to those of skill in the art. For example, the amount that is delivered is sufficient to ameliorate one or more of the symptoms of diseases or disorders associated undesired cell proliferation, coronary restenosis, osteoporosis, syndromes characterized by chronic inflammation, autoimmune diseases and

cardiovascular diseases as described herein.

Typically a therapeutically effective dosage should produce a serum concentration of active ingredient of from about 0.1 ng/ml to about 50-100 μ ml. The pharmaceutical compositions typically should provide a dosage of from about 0.001 mg to about 2000 mg of compound per kilogram of body weight per day. Pharmaceutical dosage unit forms are prepared to provide from about 1 mg to about 1000 mg and preferably from about 10 to about 500 mg of the essential active ingredient or a combination of essential ingredients per dosage unit form.

The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at intervals of time. It is understood that the precise dosage and duration of treatment is a function of the disease being treated and may be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test data. It is to be noted that concentrations and dosage values may also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions.

Pharmaceutically acceptable derivatives include acids, bases, enol ethers and esters, salts, esters, hydrates, solvates and prodrug forms. The derivative is selected such that its pharmacokinetic properties are superior to the corresponding neutral compound. Thus, effective concentrations or amounts of one or more of the compounds described herein or pharmaceutically acceptable derivatives thereof are mixed with a suitable pharmaceutical carrier or vehicle for systemic, topical or local administration to form pharmaceutical compositions. Compounds are included in an amount effective for ameliorating one or more symptoms of, or for treating or preventing diseases or disorders associated with undesired cell proliferation, coronary restenosis, osteoporosis, syndromes characterized by chronic inflammation, autoimmune diseases and

cardiovascular diseases as described herein. The concentration of active compound in the composition will depend on absorption, inactivation, excretion rates of the active compound, the dosage schedule, amount administered, particular formulation as well as other factors known to those of skill in the art.

The compositions are intended to be administered by a suitable route, including orally, parenterally, rectally, topically and locally. For oral administration, capsules and tablets are presently preferred. The

compositions are in liquid, semi-liquid or solid form and are formulated in a manner suitable for each route of administration. Preferred modes of administration include parenteral and oral modes of administration. Oral administration is presently most preferred.

Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include any of the following components: a sterile diluent, such as water for injection, saline solution, fixed oil, polyethylene glycol, glycerine, propylene glycol or other synthetic solvent; antimicrobial agents, such as benzyl alcohol and methyl parabens; antioxidants, such as ascorbic acid and sodium bisulfite; chelating agents, such as ethylenediamine- tetraacetic acid (EDTA); buffers, such as acetates, citrates and phosphates; and agents for the adjustment of tonicity such as sodium chloride or dextrose. Parenteral preparations can be enclosed in ampules, disposable syringes or single or multiple dose vials made of glass, plastic or other suitable material. In instances in which the compounds exhibit insufficient solubility, methods for solubilizing compounds may be used. Such methods are known to those of skill in this art, and include, but are not limited to, using cosolvents, such as dimethylsulfoxide (DMSO), using surfactants, such as TWEEN®, or dissolution in aqueous sodium bicarbonate.

Upon mixing or addition of the compound(s), the resulting mixture may be a solution, suspension, emulsion or the like. The form of the resulting mixture depends upon a number of factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient for ameliorating the

symptoms of the disease, disorder or condition treated and may be empirically determined.

The pharmaceutical compositions are provided for administration to humans and animals in unit dosage forms, such as tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, and oral solutions or suspensions, and oil-water emulsions containing suitable quantities of the compounds or pharmaceutically acceptable derivatives thereof. The pharmaceutically therapeutically active compounds and derivatives thereof are typically formulated and administered in unit-dosage forms or multiple-dosage forms. Unit-dose forms as used herein refers to physically discrete units suitable for human and animal subjects and packaged individually as is known in the art. Each unit-dose contains a predetermined quantity of the therapeutically active compound sufficient to produce the desired therapeutic effect, in association with the required pharmaceutical carrier, vehicle or diluent. Examples of unit-dose forms include ampules and syringes and individually packaged tablets or capsules. Unit-dose forms may be administered in fractions or multiples thereof. A multiple-dose form is a plurality of identical unit-dosage forms packaged in a single container to be administered in segregated unit-dose form. Examples of multiple-dose forms include vials, bottles of tablets or capsules or bottles of pints or gallons. Hence, multiple dose form is a multiple of unit-doses which are not segregated in packaging.

The composition can contain along with the active ingredient: a diluent such as lactose, sucrose, dicalcium phosphate, or carboxymethylcellulose; a lubricant, such as magnesium stearate, calcium stearate and talc; and a binder such as starch, natural gums, such as gum acaciagelatin, glucose, molasses, polvinylpyrrolidine, celluloses and derivatives thereof, povidone, crospovidones and other such binders known to those of skill in the art. Liquid

pharmaceutically administrable compositions can, for example, be prepared by dissolving, dispersing, or otherwise mixing an active compound as defined above and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline, aqueous dextrose, glycerol, glycols, ethanol, and the like, to thereby form a solution or suspension. If desired, the pharmaceutical composition to be administered may also contain minor amounts of nontoxic auxiliary substances such as wetting agents, emulsifying agents, or

solubilizing agents, pH buffering agents and the like, for example, acetate, sodium citrate, cyclodextrine derivatives, sorbitan monolaurate,

triethanolamine sodium acetate, triethanolamine oleate, and other such agents. Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington's

Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 15th Edition, 1975. The composition or formulation to be administered will, in any event, contain a quantity of the active compound in an amount sufficient to alleviate the symptoms of the treated subject.

Dosage forms or compositions containing active ingredient in the range of 0.005% to 100% with the balance made up from non-toxic carrier may be prepared. For oral administration, a pharmaceutically acceptable non-toxic composition is formed by the incorporation of any of the normally employed excipients, such as, for example pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, talcum, cellulose derivatives, sodium crosscarmellose, glucose, sucrose, magnesium carbonate or sodium saccharin. Such compositions include solutions, suspensions, tablets, capsules, powders and sustained release formulations, such as, but not limited to, implants and microencapsulated delivery systems, and biodegradable, biocompatible polymers, such as collagen, ethylene vinyl acetate, polyanhydrides, polyglycolic acid, polyorthoesters, polylactic acid and others. Methods for preparation of these compositions are known to those skilled in the art. The contemplated compositions may contain 0.001%-100% active ingredient, preferably 0.1-85%, typically 75-95%.

The active compounds or pharmaceutically acceptable derivatives may be prepared with carriers that protect the compound against rapid elimination from the body, such as time release formulations or coatings.

The compositions may include other active compounds to obtain desired combinations of properties. The compounds provided herein, or

pharmaceutically acceptable derivatives thereof as described herein, may also be advantageously administered for therapeutic or prophylactic purposes together with another pharmacological agent known in the general art to be of value in treating one or more of the diseases or medical conditions referred to hereinabove, such as diseases or disorders associated with undesired cell proliferation, coronary restenosis, osteoporosis, syndromes characterized by chronic inflammation, autoimmune diseases and cardiovascular diseases. It is to be understood that such combination therapy constitutes a further aspect of the compositions and methods of treatment provided herein. 1. Compositions for oral administration

Oral pharmaceutical dosage forms are solid, gel or liquid. The solid dosage forms are tablets, capsules, granules, and bulk powders. Types of oral tablets include compressed, chewable lozenges and tablets, which may be enteric-coated, sugar-coated or film-coated. Capsules may be hard or soft gelatin capsules, while granules and powders may be provided in nonOeffervescent or effervescent form with the combination of other

ingredients known to those skilled in the art.

In certain embodiments, the formulations are solid dosage forms, preferably capsules or tablets. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder; a diluent; a disintegrating agent; a lubricant; a glidant; a sweetening agent; and a flavoring agent.

Examples of binders include microcrystalline cellulose, gum tragacanth, glucose solution, acacia mucilage, gelatin solution, sucrose and starch paste. Lubricants include talc, starch, magnesium or calcium stearate, lycopodium and stearic acid. Diluents include, for example, lactose, sucrose, starch, kaolin, salt, mannitol and dicalcium phosphate. Glidants include, but are not limited to, colloidal silicon dioxide. Disintegrating agents include crosscarmellose sodium, sodium starch glycolate, alginic acid, corn starch, potato starch, bentonite, methylcellulose, agar and carboxymethylcellulose. Coloring agents include, for example, any of the approved certified water soluble FD and C dyes, mixtures thereof; and water insoluble FD and C dyes suspended on alumina hydrate. Sweetening agents include sucrose, lactose, mannitol and artificial sweetening agents such as saccharin, and any number of spray dried flavors. Flavoring agents include natural flavors extracted from plants such as fruits and synthetic blends of compounds which produce a pleasant sensation, such as, but not limited to peppermint and methyl salicylate. Wetting agents include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate and polyoxyethylene laural ether. EmeticOcoatings include fatty acids, fats, waxes, shellac, ammoniated shellac and cellulose acetate

phthalates. Film coatings include hydroxyethylcellulose, sodium

carboxymethylcellulose, polyethylene glycol 4000 and cellulose acetate phthalate.

If oral administration is desired, the compound could be provided in a composition that protects it from the acidic environment of the stomach. For example, the composition can be formulated in an enteric coating that maintains its integrity in the stomach and releases the active compound in the intestine. The composition may also be formulated in combination with an antacid or other such ingredient.

When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials which modify the physical form of the dosage unit, for example, coatings of sugar and other enteric agents. The compounds can also be administered as a component of an elixir, suspension, syrup, wafer, sprinkle, chewing gum or the like. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.

The active materials can also be mixed with other active materials which do not impair the desired action, or with materials that supplement the desired action, such as antacids, H2 blockers, and diuretics. The active ingredient is a compound or pharmaceutically acceptable derivative thereof as described herein. Higher concentrations, up to about 98% by weight of the active ingredient may be included.

Pharmaceutically acceptable carriers included in tablets are binders, lubricants, diluents, disintegrating agents, coloring agents, flavoring agents, and wetting agents. Enteric-coated tablets, because of the enteric-coating, resist the action of stomach acid and dissolve or disintegrate in the neutral or alkaline intestines. Sugar-coated tablets are compressed tablets to which different layers of pharmaceutically acceptable substances are applied. Film- coated tablets are compressed tablets which have been coated with a polymer or other suitable coating. Multiple compressed tablets are compressed tablets made by more than one compression cycle utilizing the pharmaceutically acceptable substances previously mentioned. Coloring agents may also be used in the above dosage forms. Flavoring and sweetening agents are used in compressed tablets, sugar-coated, multiple compressed and chewable tablets. Flavoring and sweetening agents are especially useful in the formation of chewable tablets and lozenges.

Liquid oral dosage forms include aqueous solutions, emulsions, suspensions, solutions and/or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules. Aqueous solutions include, for example, elixirs and syrups.

Emulsions are either oil-in-water or water-in-oil.

Elixirs are clear, sweetened, hydro alcoholic preparations.

Pharmaceutically acceptable carriers used in elixirs include solvents. Syrups are concentrated aqueous solutions of a sugar, for example, sucrose, and may contain a preservative. An emulsion is a two-phase system in which one liquid is dispersed in the form of small globules throughout another liquid.

Pharmaceutically acceptable carriers used in emulsions are non-aqueous liquids, emulsifying agents and preservatives. Suspensions use

pharmaceutically acceptable suspending agents and preservatives.

Pharmaceutically acceptable substances used in non-effervescent granules, to be reconstituted into a liquid oral dosage form, include diluents, sweeteners and wetting agents. Pharmaceutically acceptable substances used in

effervescent granules, to be reconstituted into a liquid oral dosage form, include organic acids and a source of carbon dioxide. Coloring and flavoring agents are used in all of the above dosage forms.

Solvents include glycerin, sorbitol, ethyl alcohol and syrup. Examples of preservatives include glycerin, methyl and propylparaben, benzoic add, sodium benzoate and alcohol. Examples of non-aqueous liquids utilized in emulsions include mineral oil and cottonseed oil. Examples of emulsifying agents include gelatin, acacia, tragacanth, bentonite, and surfactants such as polyoxyethylene sorbitan monooleate. Suspending agents include sodium

carboxymethylcellulose, pectin, tragacanth, Veegum and acacia. Diluents include lactose and sucrose. Sweetening agents include sucrose, syrups, glycerin and artificial sweetening agents such as saccharin. Wetting agents include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate and polyoxyethylene lauryl ether. Organic adds include citric and tartaric acid. Sources of carbon dioxide include sodium bicarbonate and sodium carbonate. Coloring agents include any of the approved certified water soluble FD and C dyes, and mixtures thereof. Flavoring agents include natural flavors extracted from plants such fruits, and synthetic blends of compounds which produce a pleasant taste sensation.

For a solid dosage form, the solution or suspension, in for example propylene carbonate, vegetable oils or triglycerides, is preferably encapsulated in a gelatin capsule. Such solutions, and the preparation and encapsulation thereof, are disclosed in U.S. Patent Nos 4,328,245; 4,409,239; and 4,410,545. For a liquid dosage form, the solution, e.g., for example, in a polyethylene glycol, may be diluted with a sufficient quantity of a pharmaceutically acceptable liquid carrier, e.g., water, to be easily measured for administration.

Alternatively, liquid or semi-solid oral formulations may be prepared by dissolving or dispersing the active compound or salt in vegetable oils, glycols, triglycerides, propylene glycol esters (e.g., propylene carbonate) and other such carriers, and encapsulating these solutions or suspensions in hard or soft gelatin capsule shells. Other useful formulations include those set forth in U.S. Patent Nos. Re 28,819 and 4,358,603. Briefly, such formulations include, but are not limited to, those containing a compound provided herein, a dialkylated mono- or poly-alkylene glycol, including, but not limited to, 1,2- dimethoxymethane, diglyme, triglyme, tetraglyme, polyethylene glycol-350- dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol- 750-dimethyl ether wherein 350, 550 and 750 refer to the approximate average molecular weight of the polyethylene glycol, and one or more antioxidants, such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone, hydroxycoumarins, ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric acid, thiodipropionic acid and its esters, and dithiocarbamates. Other formulations include, but are not limited to, aqueous alcoholic solutions including a pharmaceutically acceptable acetal. Alcohols used in these formulations are any pharmaceutically acceptable water-miscible solvents having one or more hydroxyl groups, including, but not limited to, propylene glycol and ethanol. Acetals include, but are not limited to, di(lower alkyl) acetals of lower alkyl aldehydes such as acetaldehyde diethyl acetal.

In all embodiments, tablets and capsules formulations may be coated as known by those of skill in the art in order to modify or sustain dissolution of the active ingredient. Thus, for example, they may be coated with a

conventional enterically digestible coating, such as phenylsalicylate, waxes and cellulose acetate phthalate.

2. Injectables, solutions and emulsions

Parenteral administration, generally characterized by injection, either subcutaneously, intramuscularly or intravenously is also contemplated herein. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. Suitable excipients are, for example, water, saline, dextrose, glycerol or ethanol. In addition, if desired, the pharmaceutical compositions to be administered may also contain minor amounts of nonDtoxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers, and other such agents, such as for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate and cyclodextrins. Implantation of a slowDrelease or sustainedDrelease system, such that a constant level of dosage is maintained (see, e.g., U.S. Patent No.

3,710,795) is also contemplated herein. Briefly, a compound provided herein is dispersed in a solid inner matrix, e.g., polymethylmethacrylate,

polybutylmethacrylate, plasticized or unplasticized polyvinylchloride, plasticized nylon, plasticized polyethyleneterephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene- vinylacetate copolymers, silicone rubbers, polydimethylsiloxanes, silicone carbonate copolymers, hydrophilic polymers such as hydrogels of esters of acrylic and methacrylic acid, collagen, cross-linked polyvinylalcohol and cross- linked partially hydrolyzed polyvinyl acetate, that is surrounded by an outer polymeric membrane, e.g., polyethylene, polypropylene, ethylene/propylene copolymers, ethylene/ethyl acrylate copolymers, ethylene/vinylacetate copolymers, silicone rubbers, polydimethyl siloxanes, neoprene rubber, chlorinated polyethylene, polyvinylchloride, vinylchloride copolymers with vinyl acetate, vinylidene chloride, ethylene and propylene, ionomer

polyethylene terephthalate, butyl rubber epichlorohydrin rubbers,

ethylene/vinyl alcohol copolymer, ethylene/vinyl acetate/vinyl alcohol terpolymer, and ethylene/vinyloxyethanol copolymer, that is insoluble in body fluids. The compound diffuses through the outer polymeric membrane in a release rate controlling step. The percentage of active compound contained in such parenteral compositions is highly dependent on the specific nature thereof, as well as the activity of the compound and the needs of the subject.

Parenteral administration of the compositions includes intravenous, subcutaneous and intramuscular administrations. Preparations for parenteral administration include sterile solutions ready for injection, sterile dry soluble products, such as lyophilized powders, ready to be combined with a solvent just prior to use, including hypodermic tablets, sterile suspensions ready for injection, sterile dry insoluble products ready to be combined with a vehicle just prior to use and sterile emulsions. The solutions may be either aqueous or nonaqueous.

If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, and

polypropylene glycol and mixtures thereof.

Pharmaceutically acceptable carriers used in parenteral preparations include aqueous vehicles, nonaqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents and other pharmaceutically acceptable substances.

Examples of aqueous vehicles include Sodium Chloride Injection,

Ringers Injection, Isotonic Dextrose Injection, Sterile Water Injection,

Dextrose and Lactated Ringers Injection. Nonaqueous parenteral vehicles include fixed oils of vegetable origin, cottonseed oil, corn oil, sesame oil and peanut oil. Antimicrobial agents in bacteriostatic or fungistatic concentrations must be added to parenteral preparations packaged in multiple-dose

containers which include phenols or cresols, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphate and citrate.

Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspending and dispersing agents include sodium

carboxymethylcelluose, hydroxypropyl methylcellulose and

polyvinylpyrrolidone. Emulsifying agents include Polysorbate 80 (TWEEN® 80). A sequestering or chelating agent of metal ions include EDTA.

Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol and propylene glycol for water miscible vehicles and sodium hydroxide,

hydrochloric acid, citric acid or lactic acid for pH adjustment.

The concentration of the pharmaceutically active compound is adjusted so that an injection provides an effective amount to produce the desired pharmacological effect. The exact dose depends on the age, weight and condition of the patient or animal as is known in the art.

The unit-dose parenteral preparations are packaged in an ampule, a vial or a syringe with a needle. All preparations for parenteral administration must be sterile, as is known and practiced in the art.

Illustratively, intravenous or intraarterial infusion of a sterile aqueous solution containing an active compound is an effective mode of administration. Another embodiment is a sterile aqueous or oily solution or suspension containing an active material injected as necessary to produce the desired pharmacological effect.

Injectables are designed for local and systemic administration.

Typically a therapeutically effective dosage is formulated to contain a concentration of at least about 0.1% w/w up to about 90% w/w or more, preferably more than 1% w/w of the active compound to the treated tissue(s). The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at intervals of time. It is understood that the precise dosage and duration of treatment is a function of the tissue being treated and may be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test data. It is to be noted that concentrations and dosage values may also vary with the age of the individual treated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the formulations, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed formulations.

The compound may be suspended in micronized or other suitable form or may be derivatized to produce a more soluble active product or to produce a prodrug. The form of the resulting mixture depends upon a number of factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient for ameliorating the symptoms of the condition and may be empirically determined.

3. Lyophilized powders Of interest herein are also lyophilized powders, which can be

reconstituted for administration as solutions, emulsions and other mixtures. They may also be reconstituted and formulated as solids or gels.

The sterile, lyophilized powder is prepared by dissolving a compound provided herein, or a pharmaceutically acceptable derivative thereof, in a suitable solvent. The solvent may contain an excipient which improves the stability or other pharmacological component of the powder or reconstituted solution, prepared from the powder. Excipients that may be used include, but are not limited to, dextrose, sorbital, fructose, corn syrup, xylitol, glycerin, glucose, sucrose or other suitable agent. The solvent may also contain a buffer, such as citrate, sodium or potassium phosphate or other such buffer known to those of skill in the art at, typically, about neutral pH. Subsequent sterile filtration of the solution followed by lyophilization under standard conditions known to those of skill in the art provides the desired formulation. Generally, the resulting solution will be apportioned into vials for lyophilization. Each vial will contain a single dosage (10-1000 mg, preferably 100-500 mg) or multiple dosages of the compound. The lyophilized powder can be stored under appropriate conditions, such as at about 4 °C to room temperature.

Reconstitution of this lyophilized powder with water for injection provides a formulation for use in parenteral administration. For

reconstitution, about 1-50 mg, preferably 5-35 mg, more preferably about 9-30 mg of lyophilized powder, is added per mL of sterile water or other suitable carrier. The precise amount depends upon the selected compound. Such amount can be empirically determined.

4. Topical administration

Topical mixtures are prepared as described for the local and systemic administration. The resulting mixture may be a solution, suspension, emulsions or the like and are formulated as creams, gels, ointments,

emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigations, sprays, suppositories, bandages, dermal patches or any other formulations suitable for topical administration.

The compounds or pharmaceutically acceptable derivatives thereof may be formulated as aerosols for topical application, such as by inhalation (see, e.g., U.S. Patent Nos. 4,044,126, 4,414,209, and 4,364,923, which describe aerosols for delivery of a steroid useful for treatment of inflammatory diseases, particularly asthma). These formulations for administration to the respiratory tract can be in the form of an aerosol or solution for a nebulizer, or as a microfine powder for insufflation, alone or in combination with an inert carrier such as lactose. In such a case, the particles of the formulation will typically have diameters of less than 50 microns, preferably less than 10 microns.

The compounds may be formulated for local or topical application, such as for topical application to the skin and mucous membranes, such as in the eye, in the form of gels, creams, and lotions and for application to the eye or for intracisternal or intraspinal application. Topical administration is

contemplated for transdermal delivery and also for administration to the eyes or mucosa, or for inhalation therapies. Nasal solutions of the active compound alone or in combination with other pharmaceutically acceptable excipients can also be administered.

These solutions, particularly those intended for ophthalmic use, may be formulated as 0.01% - 10% isotonic solutions, pH about 5-7, with appropriate salts.

5. Compositions for other routes of administration

Other routes of administration, such as topical application, transdermal patches, and rectal administration are also contemplated herein.

For example, pharmaceutical dosage forms for rectal administration are rectal suppositories, capsules and tablets for systemic effect. Rectal suppositories are used herein mean solid bodies for insertion into the rectum which melt or soften at body temperature releasing one or more

pharmacologically or therapeutically active ingredients. Pharmaceutically acceptable substances utilized in rectal suppositories are bases or vehicles and agents to raise the melting point. Examples of bases include cocoa butter (theobroma oil), glycerin- gelatin, carbowax (polyoxyethylene glycol) and appropriate mixtures of mono-, di- and triglycerides of fatty acids.

Combinations of the various bases may be used. Agents to raise the melting point of suppositories include spermaceti and wax. Rectal suppositories may be prepared either by the compressed method or by molding. The typical weight of a rectal suppository is about 2 to 3 gm.

Tablets and capsules for rectal administration are manufactured using the same pharmaceutically acceptable substance and by the same methods as for formulations for oral administration.

6. Articles of manufacture

The compounds or pharmaceutically acceptable derivatives thereof can be packaged as articles of manufacture containing packaging material, a compound or pharmaceutically acceptable derivative thereof provided herein, which is used for treatment, prevention or amelioration of one or more symptoms associated with undesired cell proliferation, coronary restenosis, osteoporosis, syndromes characterized by chronic inflammation, autoimmune diseases and cardiovascular diseases condition, and a label that indicates that the compound or pharmaceutically acceptable derivative thereof is used for treatment, prevention or amelioration of one or more symptoms associated with undesired cell proliferation, coronary restenosis, osteoporosis, syndromes characterized by chronic inflammation, autoimmune diseases and

cardiovascular diseases.

The articles of manufacture provided herein contain packaging materials. Packaging materials for use in packaging pharmaceutical products are well known to those of skill in the art. See, e.g., U.S. Patent Nos.

5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging material suitable for a selected formulation and intended mode of administration and treatment. A wide array of formulations of the compounds and compositions provided herein are contemplated as are a variety of treatments for any disorder associated with undesired cell proliferation, coronary restenosis, osteoporosis, syndromes characterized by chronic inflammation, autoimmune diseases and cardiovascular diseases condition.

Methods of use of the compounds and compositions

The compounds of the invention are either naturally occurring- molecules (compounds 1 and 2) or structural analogs (compounds 3 and 4, or compounds of the general formula A-K) of naturally-occurring molecules that are known to have biological activity against a wide variety of targets, including diseases or conditions associated with inflammation or inflammatory response, undesired cell proliferation, such as cancer, and cardiovascular diseases. As such, the compounds of the invention are expected to have similar activity against those targets.

It is known in the art that compounds that have a carbon-carbon double bond with the Z configuration substituted with a carbon-carbon triple bond, they tend to retain to a great extend the same biological activity as the parent compounds acid [Petasis, N. A.; Akritopoulou-Zanze, I.; Fokin, V. V.;

Bernasconi, G.; Keledjian, R.; Yang, R.; Uddin, J.; Nagulapalli, K. C; Serhan, C. N. Prostaglandins Leukot. Essent. Fatty Acids 2005, 73, 301-321]. Therefore we anticipate that the provided compounds having such alkyne bonds will also be similarly active.

Accordingly, in one aspect the invention features methods of

ameliorating or treating diseases or conditions associated with inflammation or inflammatory response, involving the administration to a subject of a therapeutically effective amount of a compound or compounds of the invention, such that inflammation or an inflammatory response are significantly reduced or eliminated in the subject. A significant reduction includes the reduction or elimination of a symptom or symptoms associated with the inflammation or inflammatory response.

In another aspect, the invention features methods of ameliorating or treating diseases or conditions associated with undesired cell proliferation, such as cancer, involving the administration to a subject of an effective amount of a compound or compounds of the invention. In general, an effective amount is an amount sufficient to ensure adequate exposure of a target cell population, such that abnormal cell proliferation is substantially slowed or halted. A target population is a population of cells undergoing abnormal cell

proliferation, such as cancerous and/or tumorous growth.

The provided methods and compositions are suitable for the treatment of a wide range of inflammatory diseases. These include but are not limited to: cardiovascular disease, arthritis, asthma, acute lung injury, chronic

obstructive pulmonary disease, cystic fibrosis, pancreatitis, systemic lupus erythematosus, Sjogren's syndrome, thyroiditis, atherosclerosis, colitis, irritable bowel disease, celiac disease, Crohn's disease, fibromyalgia, nephritis, dermatitis, acne, periodontal disease, sepsis, stroke, Alzheimer's disease, Parkinson's disease, ophthalmic inflammation, retinopathy, retinal edema, uveitis, age-related macular degeneration, diabetes, and cancer.

The invention will be further described in the following examples, which are illustrative only, and which are not intended to limit the scope of the invention described in the claims.

EXAMPLES

Specific embodiments of certain aspects of the present invention are described herein. In the following examples outlined in Figures 2 - 5, efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees centigrade, and pressure is at or near atmospheric. Starting materials used in these examples are generally either commercially available or can be readily prepared from commercially available reagents by a procedure involving one or more steps.

Example: Preparation of Compounds 1, 2 and 3

(S)-0-i-Butyldimethylsilyl glycidol (8). To a solution of TBS-C1 (15.3 g, 101.5 mmol), imidazole (6.9 g, 101.5 mmol) and DMAP (412 mg, 3.4 mmol) dissolved in 125 mL dry CH ₂ C1 ₂ at 0°C was added .R-glycidol (5.0 g, 67.5 mmol) at 0°C. The reaction was allowed to stir overnight at room temperature. It was then quenched with saturated aqueous NH4CI (125 mL) and extracted with Et2O (3 x 125 mL). The combined extract was dried with Na2SO ₄ and evaporated to give a crude clear oil which was then chromatographed on silica gel using EtOAc-hexanes mixture of (1:20) as the eluent to afford the (S)-protected glycidol 8 as a viscous and colorless oil (12.2 g, 96%). Ή NMR (400 MHz, CDCI3) δ 3.84 (dd, J= 11.9, 3.2 Hz, 1H), 3.65 (dd, J = 11.9, 4.8 Hz, 1H), 3.13 - 3.01 (m, 1H), 2.76 (dd, J= 5.2, 4.0 Hz, 1H), 2.63 (dd, J = 5.2, 2.7 Hz, 1H), 0.90 (s, 9H), 0.08 (s, 3H), 0.07 (s, 3H). C NMR (400 MHz, CDCI3) 63.88, 52.56, 44.60, 26.01, 18.50, -5.17, -5.21. (Ref. Wang, Jian-Chao; J. Org. Chem. 1999, 64, 8090-9097).

(S)-l-(i-Butyldiphenylsilyloxy)hept-4-yn-2-ol (16). To a solution of 1-butyne (286 mg, 5.3 mmol) in dry THF (10 mL) was added 2.5 M ?i-BuLi (2.12 mL, 5.3 mmol) at -78°C. After 0.25h BF ₃ .Et ₂ O (0.64 mL, 5.3 mmol) was added dropwise at -78°C. To the reaction mixture was added glycidol derivative 8 (0.5 g, 2.65 mmol) and stirred for 3 h at -78°C. The reaction mixture was warmed to room temperature, quenched with saturated aqueous NH ₄ C1 (15 mL) and extracted with Et20 (3 x 15 mL). The organic layer was dried with MgS0 ₄ , filtered and the solvent removed in vacuo. The crude reaction mixture was purified on silica gel using EtOAc-hexanes (4%) as the eluent to afford compound 16 as a clear colorless oil (600 mg, 94%). Ή NMR (400 MHz, CDCls) δ 3.77 - 3.72 (m, 1H), 3.70 (dd, J = 9.9, 4.2 Hz, 1H), 3.60 (dd, J = 9.8, 5.8 Hz, 1H), 2.46 (d, J = 4.8 Hz, 1H), 2.42 - 2.33 (m, 2H), 2.22 - 2.10 (m, 2H), 1.11 (t, 3H), 0.90 (s, 9H), 0.08 (s, 6H). ⁱ »C NMR (400 MHz, CDC1 ₃ ) δ 84.15, 75.21, 70.63, 65.81, 26.03, 23.55, 18.46, 14.33, 12.55, -5.23, -5.26.

(S)-2-(i-Butyldiphenylsilyloxy)hept-4-yn-l-ol (17) Part 1: To a flask with imidazole (95 mg, 1.39 mmol) and DMAP (8 mg, 0.06 mmol) in CH ₂ C1 ₂ (5 mL total volume) was added TBDPS-C1 (0.36 mL, 1.39 mmol) dropwise at 0°C. The alcohol 16 (280 mg, 1.15 mmol) was cannulated to the flask and stirred overnight at room temperature. The reaction mixture was quenched with saturated aqueous NH ₄ C1 (7 mL) and extracted with Et20 (3 x 7 mL). The organic layer was dried with MgS0 ₄ , filtered and the solvent removed in vacuo. The crude reaction mixture was purified on silica gel using EtOAc- hexanes (1%) as the eluent to afford di-protected derivative (Sj-l-ii- butyldimethylsilyloxy)-2-(i-butyldiphenylsilyloxy)hept-4-yne (525 mg, 89%) as a clear colorless oil. Ή NMR (400 MHz, CDCI3) δ 7.85 - 7.65 (m, 4H), 7.55 - 7.29 (m, 6H), 3.83 (p, J= 5.5 Hz, 1H), 3.55 (dd, J= 5.4, 3.3 Hz, 2H), 2.44 - 2.34 (m, 1H), 2.32 - 2.23 (m, 1H), 2.12 (qt, J = 7.5, 2.4 Hz, 2H), 1.10 (t, 3H), 1.07 (s, 9H), 0.84 (s, 9H), -0.03 (s, 3H), -0.06 (s, 3H). ¹³ C NMR (400 MHz, CDCI3) δ 136.10, 136.03, 134.47, 134.34, 129.69, 129.66, 127.62, 83.29, 76.47, 72.78, 65.80, 27.09, 26.05, 24.12, 19.54, 18.46, 14.34, 12.62, -5.34, -5.35. Part 2: To a solution of the product of Part 1 (0.5 g, 1.04 mmol) in CH2CI2 (5 mL) was added camphor sulfonic acid (144 mg, 0.62 mmol) at room temperature and monitored for lh. The reaction was quenched with Et3N (0.09 mL, 0.62 mmol) and the solvent was evaporated in vacuo without workup. The crude reaction mixture was purified on silica gel using EtOAc-hexanes (1:10) as the eluent to afford alcohol 17 as a clear colorless oil (370 mg, 97%). Ή NMR (400 MHz, CDC1 ₃ ) δ 7.82 - 7.72 (m, 4H), 7.57 - 7.37 (m, 6H), 4.06 - 3.87 (m, 1H), 3.70 (d, J = 4.6 Hz, 2H), 2.47 (ddt, J= 16.4, 7.7, 2.5 Hz, 1H), 2.36 (ddt, J= 16.4, 4.9, 2.4 Hz, 1H), 2.18 - 2.06 (m, 3H), 1.16 (s, 9H), 1.10 (t, J= 7.5 Hz, 3H). NMR (400 MHz, CDCI3) δ 135.86, 135.71, 133.64, 133.62, 129.89, 129.85, 127.80, 127.68, 83.83, 75.43, 72.62, 65.56, 27.01, 23.91, 19.33, 14.10, 12.4. (2S, 4 )-2-(i-Butyldiphenylsilyloxy)hept-4-en-l-ol (18). To a solution of alcohol 17 (3.9 g, 10.2 mmol) in EtOAc (200 mL) was added Lindlar catalyst (200 mg). The reaction mixture was placed under a ¾ atmosphere and stirred for 2 h. The reaction was filtered through celite and the solvent was removed in vacuo. The crude product was purified on silica gel using EtOAc-hexanes (3:22) as an eluent to afford the alcohol 18 as a clear colorless oil (3.5 g, 94%). 1H NMR (400 MHz, CDCI3) δ 7.89 - 7.56 (m, 4H), 7.51 - 7.30 (m, 6H), 5.43 - 5.28 (m, 1H), 5.17 (dtt, J = 10.7, 7.6, 1.5 Hz, 1H), 3.79 (dtd, J = 8.5, 4.8, 3.6 Hz, 1H), 3.63 - 3.43 (m, 2H), 2.36 - 2.23 (m, 1H), 2.22 - 2.10 (m, 1H), 1.91 - 1.73 (m, 2H), 1.09 (s, 9H), 0.84 (t, J = 7.5 Hz, 3H). ¹³ C NMR (400 MHz, CDCI3) δ 136.03, 135.85, 134.42, 133.97, 133.93, 129.97, 129.93, 127.91, 127.80, 123.7 , 74.00, 65.74, 31.69, 27.19, 20.61, 19.49, 14.24.

(3S, IE, 5 )-3-(i-Butyldiphenylsilyloxy)-l-iodoocta-l,5-diene (19). Part 1: To a solution of DMSO (0.43 mL, 3.7 mmol) in CH2CI2 (15 mL) was added oxalyl chloride (0.33 mL, 3.7 mmol) at -78°C. To the reaction mixture was

cannulated alcohol 18 (0.5 g, 1.87 mmol) and stirred for 1 h. Et3N was added to the reaction at -78°C and stirred for 3 h. The reaction mixture was quenched with saturated aqueous NH4CI (20 mL) and extracted with Et20 (3 x 20 mL). The Et20 was washed with brine, dried with MgS0 ₄ and filtered. The solvent was removed in vacuo and the crude product was purified on silica gel using EtOAc-Hexanes (1:4) as the eluent to afford the aldehyde 2S, 4Z, 2-(t- butyldiphenylsilyloxy)hept-4-enal as a colorless oil (0.44 g, 90%). 1H NMR (400 MHz, CDCls) δ 9.57 (d, J = 1.7 Hz, 2H), 7.71 - 7.61 (m, 4H), 7.49 - 7.33 (m, 6H), 5.52 - 5.41 (m, 1H), 5.40 - 5.27 (m, 1H), 4.06 (td, J= 6.5, 1.7 Hz, 1H), 2.44 (dt, J = 14.1, 6.7 Hz, 1H), 2.34 (dt, J= 13.9, 6.5 Hz, 1H), 1.99 - 1.87 (m, 2H), 1.12 (s, 9H), 0.90 (t, J = 7.5 Hz, 3H). «0 NMR (400 MHz, CDCI3) δ 203.52, 135.95, 135.95, 135.11, 133.24, 133.12, 130.18, 130.13, 127.95, 127.89, 122.17, 77.94, 31.15, 27.08, 20.74, 19.49, 14.14. Part 2. To a solution of CrCl ₂ (3.4 g, 27.3 mmol) dissolved in THF (40 mL total volume) was cannulated a mixture of 2S, 4Z, 2-(i-butyldiphenylsilyloxy)hept-4-enal aldehyde (1.0 g, 2.7 mmol) and CHI3 (5.4 g, 13.7 mmol) dissolved in anhydrous THF (10 mL) under Argon at 0°C. The reaction was stirred at 0°C for 3 h and an additional 1 h at room temperature. The reaction mixture was quenched with water (50 mL) extracted with Et20 (3 x 50 mL) rinsed with brine and dried over MgS0 ₄ . The organic phase was filtered and the solvent was removed in vacuo to afford a crude oil which was purified on silica gel using first pure pentanes and then EtOAc-hexanes (1:24) as the eluent to afford compound 19 as a clear colorless oil (780 mg, 59%). Ή NMR (400 MHz, CDCI3) δ 7.76 - 7.54 (m, 4H), 7.51 - 7.32 (m, 6H), 6.49 (dd, J = 14.4, 6.5 Hz, lH), 5.98 (dd, J = 14.4, 1.1 Hz, 1H), 5.49 - 5.32 (m, lH), 5.32 - 5.13 (m, lH), 4.17 - 3.98 (m, OH), 2.32 - 2.07 (m, 2H), 1.84 (dtd, J= 9.2, 7.4, 5.6 Hz, 2H), 1.07 (s, 9H), 0.87 (t, J = 7.5 Hz, 3H). ¹³ C NMR (400 MHz, CDCI3) δ 148.04, 136.03, 136.01, 134.54, 133.97, 133.65, 129.90, 129.87, 127.74, 127.72, 123.18, 76.88, 75.93, 35.31, 27.14, 20.75, 19.48, 14.33. (3S, IE, 5 >3-(i-Butyldimethylsilyloxy)-l-iodoocta-l,5-diene (6). To a solution of the TBDPS-protected vinyl iodide 19 (0.8 g, 1.63 mmol) in THF (5 mL) was added 1.0 M solution of TBAF (1.63 mL, 1.63 mmol) at 0°C and stirred for 2 h. The reaction was quenched with water (10 mL) and extracted with Et20 (5 x 10 mL), rinsed with brine, dried over MgS0 ₄ and filtered. The crude reaction mixture was purified on silica gel using EtOAc-hexanes (10%) as the eluent to afford the alcohol (3S, IE, 5Z)-l-iodoocta-l,5-diene-3-ol (0.38 g, 92%) as a clear colorless oil. Ή NMR (400 MHz, CDCls) δ 6.60 (dd, J= 14.4, 5.8 Hz, 1H), 6.37 (dd, J = 14.5, 1.3 Hz, 1H), 5.73 - 5.52 (m, 1H), 5.45 - 5.25 (m, 1H), 4.23 - 3.98 (m, 1H), 2.31 (ddd, J = 7.7, 6.5, 1.6 Hz, 2H), 2.14 - 1.95 (m, 2H), 0.98 (t, J = 7.5 Hz, 3H). ¹³ C NMR (400 MHz, CDCls) δ 147.92, 134.93, 122.82, 77.48, 77.38, 77.16, 76.84, 74.01, 34.68, 20.89, 14.35. The alcohol (0.38 g, 1.5 mmol) was protected using TBS-OTf (0.54 mL, 3.0 mmol) and 2,6-lutidine (0.52 mL, 4.5 mmol) in anhydrous CH2CI2 (20 mL). The reaction mixture was quenched after 2 h with saturated aqueous NH4CI (20 mL) and extracted with Et20 (3 x 20 mL). The Et20 was rinsed with brine, dried with MgS0 ₄ and filtered. The solvent was removed in vacuo and the crude product was purified on silica gel using EtOAc-Hexanes (1:49) as the eluent to afford the silylated vinyl iodide 5 as a colorless oil (0.54 g, 98%). ¹ H NMR (400 MHz, CDCls) δ 6.54 (dd, J = 14.3, 5.7 Hz, 1H), 6.21 (dd, J= 14.3, 1.5 Hz, 1H), 5.54 - 5.42 (m, 1H), 5.37 - 5.24 (m, 1H), 4.17 - 4.00 (m, 1H), 2.33 - 2.18 (m, 2H), 2.03 (dt, J = 14.9, 7.4 Hz, 1H), 0.96 (t, J = 7.5 Hz, 4H), 0.89 (s, 9H). C NMR (400 MHz, CDCls) δ 148.90, 134.39, 123.75, 75.86, 75.20, 35.70, 25.96, 20.90, 18.37, 14.37, -4.48, -4.68.

(R)-l, 2-di-(i-Butyldimethylsilyloxy)-5-trimethylsilyl-pent-4-yne (20). Part 1: To a solution of TMS acetylene (5.6 mL, 39.8 mmol) in THF (40 mL) was added 2.5 M 7 -BuLi (15.9 mL, 39.8 mmol) at -78°C. After 0.25 h BF ₃ .Et ₂ 0 (5.0 mL, 39.8 mmol) was added drop wise at -78°C. To the reaction mixture was added (jR)-O-i-butyldimethylsilyl glycidol 13 (5.0 g, 26.5 mmol) and stirred for 3 h at - 78°C. The reaction mixture was warmed to room temperature, quenched with saturated aqueous NH4CI (45 mL) and extracted with Et20 (3 x 45 mL). The organic layer was dried with MgS0 ₄ , filtered and the solvent removed in vacuo. The crude reaction mixture was purified on silica gel using EtOAc- hexanes (4%) as the eluent to afford 2R, 1, (i-butyldimethyl-silyloxy)-5- trimethylsilyl-pent-4-yn-2-ol (5.5 g, 73%) as a clear colorless oil. ^X H NMR (400 MHz, CDCls) δ 3.78 (dt, J = 11.1, 5.9 Hz, 1H), 3.72 (dd, J = 10.0, 4.1 Hz, 1H), 3.63 (dd, J = 10.0, 5.6 Hz, 1H), 2.51 - 2.41 (m, 2H), 1.64 (s, lH), 0.91 (s, 11H), 0.14 (s, 2H), 0.09 (s, 5H), 0.08 (s, 6H). C NMR (400 MHz, CDCls) δ 102.96, 87.19, 70.32, 65.60, 26.03, 24.67, 18.48, 0.17, -5.24, -5.28. Part 2: To a solution of the product of Part 1 (5.7g, 19.9 mmol) in anhydrous CH2CI2 (30 mL) was added TBS-C1 (3.6 g, 23.9 mmol), imidazole (1.6g, 23.9 mmol), and DMAP (243 mg, 1.99 mmol) at 0°C. The reaction was allowed to stir at room temperature overnight. It was then quenched with saturated aqueous NH4CI (30 mL) and extracted with EtzO (3 x 30 mL). The combined extract was dried with MgSO ₄ , filtered and evaporated to give a crude clear oil which was then chromatographed on silica gel using EtOAc-hexanes mixture of (1:24) as the eluent to afford the product 20 as a viscous and colorless oil (7.5g, 94%). Ή NMR (400 MHz, CDCls) δ 3.86 - 3.75 (m, lH), 3.60 - 3.47 (m, 2H), 2.50 (dd, J = 16.8, 5.3 Hz, 1H), 2.30 (dd, J = 16.8, 6.5 Hz, 1H), 0.89 (s, 18H), 0.14 (s, 9H), 0.11 (s, 3H), 0.08 (s, 3H), 0.05 (s, 3H), 0.02 (s, 3H). C NMR (400 MHz, CDCls) δ 104.86, 85.95, 72.25, 66.83, 26.13, 26.01, 18.53, 18.29, 0.23, -2.78, -4.29, -4.44, -5.18, -5.24.

(¾j-2-(i-Butyldimethylsilyloxy)-5-trimethylsilyl-pent-4- yn-l-ol (11). To a solution of protected diol 20 (3.4 g, 8.5 mmol) in a 1:1 mixture of dry

CHzC MeOH (20 mL) was added camphor sulfonic acid (1.0 g, 4.2 mmol) at 0°C. The reaction was allowed to stir for 0.5 h and quenched with EtsN (1.2 mL, 8.5 mmol). The solvent was removed in vacuo without work-up and purified on silica gel using EtOAc-hexanes mixture (7%) as the eluent to afford the product 11 as a viscous and colorless oil (1.9 g, 77%). Ή NMR (400 MHz, CDCls) δ 4.13 - 3.84 (m, 1H), 3.67 (ddd, J= 11.2, 5.8, 3.8 Hz, lH), 3.57 (ddd, J = 11.2, 5.8, 3.8 Hz, 1H), 2.51 - 2.33 (m, 3H), 1.88 (dd, J = 7.1, 6.0 Hz, 2H), 0.90 (s, 9H), 0.14 (s, 9H), 0.13 (s, 4H), 0.11 (s, 4H). C NMR (400 MHz, CDCI3) δ 103.47, 86.91, 71.64, 66.04, 25.94, 25.52, 18.20, 0.16, -4.34, -4.59. (¾ 2-(^Butyldimethylsilyloxy)-5-trimethylsilyl-pent-4-ynal (21). To a -78°C solution of DMSO (1.3 mL, 18.5 mmol) in anhydrous CH2CI2 (40 mL) was added oxalyl chloride (1.1 mL, 12.3 mmol) drop wise. After 0.25 h alcohol 11 (1.58 g, 6.2 mmol) was added and stirred for 1 h at -78°C. To the reaction mixture was added EtaN (4.3 mL, 30.8 mmol) and stirred for another 3 h at - 78°C. The reaction mixture was allowed to warm to room temperature and quenched with saturated aqueous NH4CI (45 mL) and extracted with Et20 (3 x 45 mL). The combined extract was dried with Na2S0 ₄ and evaporated to give a crude clear oil which was then chromatographed on silica gel using EtOAc- hexanes mixture of (1:20) as the eluent to afford the aldehyde 21 as a viscous and colorless oil (1.6g, 92%). Ή NMR (400 MHz, CDCI3) δ 9.63 (d, J = 1.3 Hz, 1H), 4.17 - 4.05 (m, 1H), 2.64 (dd, J = 17.0, 5.0 Hz, 1H), 2.48 (dd, J = 17.0, 7.8 Hz, 1H), 0.94 (s, 9H), 0.14 (s, 12H), 0.13 (s, 3H). C NMR (400 MHz, CDCI3) δ 202.25, 101.99, 87.54, 76.25, 31.08, 25.87, 24.65, 18.39, 0.08, -4.57. (2E, 4i?)-4-(i-Butyldimethylsilyloxy)-7trimethylsilyl-hept-2-en-6 -ynal (22). To a solution of aldehyde 21 (700 mg, 2.46 mmol) dissolved in Toluene ( 15 mL) was added (Triphenylphosphoranylidene)acetaldehyde (750 mg, 2.46 mmol) and heated to reflux overnight. The solvent was removed in vacuo without work up and the crude mixture was purified on silica gel using a EtOAc- hexanes mixture of (1:20) as the eluent to afford aldehyde 22 as a clear viscous oil (634 mg, 83%). Ή NMR (400 MHz, CDCI3) δ 9.60 (d, J= 8.0 Hz, 1H), 6.90 (dd, J = 15.5, 4.3 Hz, 1H), 6.31 (ddd, J= 15.5, 8.0, 1.4 Hz, 1H), 4.54 (ddd, J = 11.8, 6.1, 1.4 Hz, 1H), 2.55 (dd, J = 16.6, 6.2 Hz, 1H), 2.44 (dd, J = 16.6, 7.4 Hz, 1H), 0.91 (s, 9H), 0.15 (s, 9H), 0.11 (s, 3H), 0.07 (s, 3H). NMR (400 MHz, CDCI3) δ 193.56, 157.92, 131.40, 102.21, 70.66, 29.11, 25.87, 18.30, 0.11, -4.63, -4.69.

(3S, IE, 4iJ)-3-(i-Butyldimethylsilyloxy)-l-iodoocta-l,5-diene (10). To a solution of CrCl2 (2.0 g, 16.1 mmol) dissolved in THF (65 mL total volume) was cannulated a mixture of aldehyde 22 (560 mg, 1.8 mmol) and CHI3 (3.2 g, 8.05 mmol) dissolved in anhydrous THF (10 mL) under Argon at 0°C. The reaction was stirred at 0°C for 3 h and an additional 1 h at room temperature. The reaction mixture was quenched with water (80 mL) extracted with Et20 (3 x 80 mL) rinsed with brine and dried over MgS0 ₄ . The organic phase was filtered and the solvent was removed in vacuo to afford a crude oil which was purified on silica gel using first pure pentanes and then EtOAc-hexanes (1:24) as the eluent to afford a 9:1 mixture of compound 10 and its E, Z stereoisomer (see inset) as a clear colorless oil (660 mg, 84%). The desired product was able to be isolated but not without some loss in the yield. Ή NMR (400 MHz, CDC1 ₃ ) δ 7.02 (dd, J = 14.5, 10.7 Hz, 1H), 6.32 (d, J= 14.5 Hz, 1H), 6.15 (ddd, J = 15.2, 10.8, 1.2 Hz, 1H), 5.78 (dd, J = 15.2, 5.7 Hz, 1H), 4.27 (d, J = 6.6 Hz, 1H), 2.49 - 2.29 (m, 2H), 0.90 (s, 9H), 0.14 (s, 9H), 0.09 (s, 3H), 0.05 (s, 3H). ⁱ sC NMR (400 MHz, CDCI ₃ ) δ 144.72, 136.49, 129.69, 103.74, 86.78, 79.23, 71.48, 29.90, 25.97, 18.39, 0.20, -4.42, -4.60. (3S, IE, 4 )-3-(i-Butyldimethylsilyloxy)-l-iodoocta-l,5-diene. Ή NMR (400 MHz, CDCls) δ 6.73 (dd, J = 10.0, 7.6 Hz, 1H), 6.45 (ddt, J= 15.2, 9.9, 1.3 Hz, 1H), 6.27 (d, J= 7.6 Hz, 1H), 6.04 (dd, J = 15.2, 5.3 Hz, 1H), 4.36 (dt, J = 6.9, 5.3 Hz, 1H), 2.45 (d, J= 6.6 Hz, 2H), 0.94 (s, 9H), 0.15 (d, J = 0.5 Hz, 9H), 0.12 (s, 3H), 0.10 (d, J = 0.5 Hz, 3H). C NMR (400 MHz, CDCls) δ 140.18, 137.85, 129.99, 128.48, 86.83, 82.57, 71.65, 29.80, 26.04, 18.39, 0.24, -4.31, -4.42;

HRMS (ESI) m/z calcd for Ci ₇ H ₃ 5lNOSi ₂ : [M+NH4] ⁺ : 452.1297 found:

452.1302.

Dimethyl (2S)-2-hydroxypentanedioate (23). To a solution of (£?)-□ -carboxy butyrolactone [S-5-oxo-2-tetrahydrofurancarboxylic acid] (5.0g, 38.5 mmol) in dry MeOH (40 mL) was added 4 drops of concentrated HC1 and heated to reflux overnight. The reaction was cooled to 0°C and quenched with NaHCOe and filtered. The solvent was then evaporated with no further workup and the crude was purified on silica gel using EtOAc-hexanes mixture of (1:1) as the eluent to afford the product 23 as a viscous and colorless oil (6.5g, 97%). Ή NMR (400 MHz, CDC1 ₃ ) δ 4.23 (dd, J= 7.9, 4.2 Hz, 1H), 3.78 (s, 3H), 3.66 (s, 3H), 2.94 (s, 1H), 2.60 - 2.33 (m, 2H), 2.24 - 2.05 (m, 1H), 1.99 - 1.83 (m, 1H). ⁱ sC NMR (400 MHz, CDC1 ₃ ) δ 175.14, 173.67, 69.58, 52.76, 51.84, 29.53, 29.39.

Methyl (4S -4,5-dihydroxypentanoate (24). To a solution of compound 23 (6.5g, 37 mmol) in dry THF (55 mL) was added BH3.DMS dropwise maintaining a temperature of 10-15°C. After stirring at this temperature for 1 h, a catalytic amount of NaBH ₄ (70mg, 1.9 mmol) was added and stirred for an additional 1 h maintaining the temperature around 10°C. The reaction mixture was quenched with dry MeOH (20 mL) and stirred for an additional .5 h. The solvent was removed in vacuo with no further workup and the crude was purified on silica gel using EtOAc as the eluent to afford the diol 24 as viscous colorless oil (4.7g, 86%). Ή NMR (400 MHz, CDCI3) δ 3.73 - 3.68 (m, 1H), 3.67 (s, 3H), 3.62 (dd, J = 11.1, 3.1 Hz, 1H), 3.44 (dd, J = 11.2, 7.1 Hz, 1H), 3.20 (s, 1H), 2.88 (s, 1H), 2.47 (td, J = 7.2, 2.9 Hz, 2H), 1.84 - 1.67 (m, 2H). NMR (400 MHz, CDCI3) δ 174.74, 71.53, 66.61, 51.92, 30.38, 28.06.

Methyl (4S)-4-(i-butyldimethylsilyloxy)-5-hydroxypentanoate (14). Part 1: To a flask with stir bar was added TBS-Cl (8.2g, 54.5 mmol), imidazole (3.7g, 54.5 mmol) and DMAP (110 mg, 0.9 mmol) and dissolved in 10 mL dry DMF at 0°C. Diol 24 (2.5g, 18 mmol) was suspended in 5 mL dry DMF and cannulated into the reaction flask at 0°C. The reaction was allowed to stir overnight at room temperature. It was then quenched with saturated aqueous NH4CI (20 mL) and extracted with Et20 (3 x 20 mL). The combined extract was dried with Na2S0 ₄ and evaporated to give a crude clear oil which was then

chromatographed on silica gel using EtOAc-hexanes mixture of (1:24) as the eluent to afford methyl (4S)-4,5-6is(i-butyldimethylsilyloxy)pentanoate as a viscous and colorless oil (6.3g, 92%). Ή NMR (400 MHz, CDCI3) δ 3.74 - 3.68 (m, 1H), 3.66 (s, 3H), 3.53 (dd, J = 10.0, 5.2 Hz, 1H), 3.39 (dd, J = 10.0, 6.8 Hz, 1H), 2.52 - 2.28 (m, 2H), 2.00 - 1.86 (m, 1H), 1.79 - 1.64 (m, 1H), 0.89 (s, 9H), 0.88 (s, 9H), 0.05 (s, 3H), 0.05 (s, 3H), 0.05 (s, 3H), 0.04 (s, 3H). «0 NMR (400 MHz, CDCls) δ 174.47, 72.05, 67.14, 51.61, 29.75, 29.47, 26.10, 26.01, 18.50, 18.25, -4.18, -4.70, -5.20, -5.24. Part 2: To a solution of the product from Part 1 (7.7g, 20.5mmol) in a 1:1 mixture of dry CH ₂ Cl ₂ /MeOH (50 mL) was added camphor sulfonic acid (3.8g, 16.38 mmol) at 0°C. The reaction was allowed to stir for 0.5 h and quenched with Et3N (2.85mL, 20.5 mmol). Avoiding workup, the solvent was removed in vacuo and purified on silica gel using EtOAc- hexanes mixture (3:7) as the eluent to afford the alcohol 14 as a viscous and colorless oil (3.1g, 58%). Ή NMR (400 MHz, CDCI3) δ 3.85 - 3.77 (m, 1H), 3.67 (s, 3H), 3.59 - 3.48 (m, 1H), 3.52 - 3.41 (m, lH), 2.47 - 2.29 (m, 2H), 1.94 (t, J = 6.3 Hz, 1H), 1.92 - 1.77 (m, 2H), 0.90 (s, 9H), 0.08 (s, 6H). C NMR (400

MHz, CDCI3) δ 174.25, 71.59, 66.01, 51.78, 29.56, 28.70, 25.95, 18.20, -4.44, -4.55; HRMS (ESI) m/z calcd for C ₁₂ H270 ₄ Si : [M+H] ⁺ : 263.1673 found:

263.1673.

Methyl (4-S)-4-(i-butyldimethylsilyloxy)-5-oxopentanoate (25). To a -78°C solution of DMSO (1.32 mL, 17.1 mmol) in anhydrous CH2CI2 (30 mL) was added oxalyl chloride (1.01 mL, 11.4 mmol) dropwise. After 0.25 h alcohol 14 (1.5g, 5.7 mmol) was added and stirred for 1 h at -78°C. To the reaction mixture was added Et ₃ N (3.9 mL, 28.5 mmol) and stirred for another 3 h at - 78°C. The reaction mixture was allowed to warm to room temperature and quenched with saturated aqueous NH4CI (35 mL) and extracted with Et ₂ O (3 x 35 mL). The combined extract was dried with Na ₂ S0 ₄ and evaporated to give a crude clear oil which was then chromatographed on silica gel using EtOAc- hexanes mixture of (1:5) as the eluent to afford the aldehyde 25 as viscous colorless oil (1.4g, 98%). 1H NMR (400 MHz, CDC1 ₃ ) δ 9.59 (d, J = 1.3 Hz, 1H), 4.11 - 4.02 (m, 1H), 3.67 (s, 3H), 2.47 - 2.38 (m, 2H), 2.09 - 1.96 (m, 1H), 1.91 (td, J = 14.1, 7.6 Hz, 1H), 0.92 (s, 9H), 0.09 (s, 3H), 0.07 (s, 3H). ⁱ³ C NMR (400 MHz, CDCI3) δ 203.46, 173.42, 76.48, 51.81, 28.91, 27.58, 25.85, 18.30, -4.53, - 4.93. f4S)-Methyl-4-(i-butyldimethylsilyloxy)-hex-5-ynoate (9). Part 1: To a solution of CBr ₄ (385 mg, 1.16 mmol) at 0°C in anhydrous CH2CI2 (25 mL total volume) was cannulated PPh.3 (607 mg, 2.32 mmol) to give a clear yellow solution. To the reaction mixture at 0°C was added aldehyde 15 (150 mg, 0.58 mmol). The reaction was run for 1 h to assure completion. Without workup the solvent was evaporated in vacuo and the crude mixture was purified on silica gel using EtOAc-hexanes mixture of (1:9) as the eluent to afford the dibromo ester [methyl (4S)-4-(i-butyldimethylsilyloxy)-6,6-dibromohex-5- enoate] as a viscous and colorless oil (193 mg, 80%). 1H NMR (400 MHz, CDCls) δ 6.37 (d, J = 8.0 Hz, 1H), 4.36 (dt, J= 8.0, 6.4 Hz, 1H), 3.68 (s, 3H), 2.44 - 2.35 (m, 2H), 1.90 - 1.80 (m, 2H), 0.88 (s, 9H), 0.07 (s, 3H), 0.06 (s, 3H). ¹³ C NMR (400 MHz, CDCls) δ 173.70, 141.43, 89.11, 72.62, 51.79, 31.70, 29.57, 25.89, 18.18, -4.39, -4.91. Part 2: To a solution of dibromo ester from Part 1 (548 mg, 1.32 mmol) at -78°C in anhydrous THF (25 mL) was added 2.0 M solution of LDA (1.98 mL, 4.0 mmol) drop wise and stirred for 0.5 h. The reaction was quenched with water (30 mL) and extracted with Et ₂ 0 (3 x 30 mL), dried using MgS0 ₄ , filtered and concentrated. The crude was then purified using silica gel with a EtOAc-hexanes eluent of (3:47) to afford the alkyne product 9 (233 mg, 69%). Ή NMR (400 MHz, CDCls) δ 4.45 (td, J= 6.1, 2.1 Hz, 1H), 3.67 (s, 3H), 2.58 - 2.45 (m, 2H), 2.39 (d, J = 2.1 Hz, 1H), 2.05 - 1.94 (m, 2H), 0.90 (s, 9H), 0.13 (s, 3H), 0.10 (s, 3H). NMR (400 MHz, CDCls) δ 173.83, 84.78, 72.74, 61.71, 51.73, 33.50, 29.55, 25.87, 18.31, -4.52, - 5.02; HRMS (ESI) m/z calcd for CisHsseOsSi : [M+H] ⁺ : 257.1568 found:

257.1566. Methyl (4S, 11R, 7E, i>S 4,ll-bis(t-butyldimethylsilyloxy)-14-(trimethylsilyl)- tetradeca-7,9-dien-5,13-diynoate (26). To the arm of a three-necked flask was charged Pd(PPh.3) ₄ (45 mg, .039 mmol) and Cul (10 mg, .078 mmol) under Argon. A solution of alkyne 9 (100 mg, 0.39 mmol), and the dienyl iodide 10 mixture (191mg, 0.43 mmol) and EteN (0.64 mL, 3.9 mmol) in CeHe (3 mL) was cannulated into the reaction vessel. The reaction flask was then freeze-thawed with liquid nitrogen three times to remove oxygen. After removing any oxygen from the reaction flask the Pd(PPh.3)4 and Cul was added and the reaction mixture was stirred overnight at room temperature. The reaction was worked up with aqueous saturated NH4CI (5 mL) and extracted with Et20 (3 x 5 mL). The solvent was evaporated and the mixture was purified on silica gel using EtOAc-hexanes (1:24) as the eluent to afford compound 26 as a clear oil (202 mg, 92%). The E, Z stereoisomer could be isolated during the separation. Ή NMR (400 MHz, CDCI3) δ 6.53 (dd, J = 15.6, 10.9 Hz, 1H), 6.23 (dd, J = 15.2, 10.9 Hz, lH), 5.83 (dd, J = 15.3, 5.9 Hz, 1H), 5.58 (d, J = 15.5 Hz, 1H), 4.58 (td, J = 6.2, 1.8 Hz, 1H), 4.32 (q, J= 6.5 Hz, 1H), 3.67 (s, 3H), 2.49 (td, J = 7.6, 1.4 Hz, 2H), 2.48 - 2.29 (m, 2H), 2.07 - 1.94 (m, 2H), 0.90 (s, 18H), 0.14 (s, 9H), 0.13 (s, 3H), 0.10 (s, 3H), 0.09 (s, 3H), 0.05 (s, 3H). NMR (400 MHz, CDCI3) δ 173.94, 141.17, 138.13, 129.15, 110.92, 103.84, 92.70, 86.72, 83.92, 71.78, 62.50, 51.70, 33.65, 30.04, 29.75, 25.97, 25.95, 18.39, 18.36, 0.20, -4.37, -4.40, - 4.60, -4.92.

Methyl (4S, 11R, 7E, 5J- )-4,ll-bis(t-butyldimethylsilyloxy)tetradeca-7,9-dien- 5,13-diynoate (5). To a solution of compound 26 (80 mg, 0.14 mmol) in MeOH (3 mL) was added Na2C03 (20 mg) and stirred overnight at room temperature. The solvent was removed in vacuo and the crude was dissolved in water (5 mL) and extracted with Et20 (3 x 5 mL). The oil was purified on silica gel using EtOAc-hexanes (1:49) as the eluent to afford alkyne 4 as a colorless oil (63 mg, 91%). NMR (500 MHz, CDCI3) δ 6.54 (dd, J= 15.4, 11.0 Hz, 1H), 6.26 (dd, J = 15.1, 11.1 Hz, 1H), 5.86 (dd, J = 15.3, 5.7 Hz, 1H), 5.60 (dd, J = 15.6, 1.6 Hz, 1H), 4.58 (td, J = 6.0, 1.6 Hz, 1H), 4.42 - 4.30 (m, 1H), 3.67 (s, 3H), 2.49 (td, J = 7.2, 1.8 Hz, 2H), 2.48 - 2.38 (m, 1H), 2.33 (ddd, J = 16.4, 7.1, 2.7 Hz, 1H), 2.06 - 1.94 (m, 2H), 1.99 (t, J= 2.7 Hz, 1H), 0.90 (s, 18H), 0.13 (s, 3H), 0.10 (s, 3H), 0.08 (s, 3H), 0.05 (s, 3H). NMR (500 MHz, CDCI3) δ 173.93, 141.08, 137.75, 129.35, 111.12, 92.78, 83.87, 81.04, 71.49, 70.37, 62.48, 51.72, 33.66, 29.75, 28.59, 25.95, 25.94, 18.39, 18.36, -4.35, -4.44, -4.68, -4.92; HRMS (ESI) m/z calcd for C ₂ 7H46Na0 ₄ Si2 : [M+Na] ⁺ : 513.2827 found: 513.2838.

Methyl (4S, 11R, 17 S, 7E, 9E, 15E, 2SZ ins-(t-butyldimethylsilyloxy)-docosa- 7,9,15,19-tetraen-5,13-diynoate (27). To the arm of a three-necked flask was charged Pd(PPh.3)4 (5 mg, .0039 mmol) and Cul (1 mg, .078 mmol) under Argon. A solution of alkyne 4 (19 mg, 0.039 mmol), vinyl iodide 5 (21 mg, 0.058 mmol) and EteN (0.05 mL, .39 mmol) in CeHe (1 mL) was cannulated into the reaction vessel. The reaction flask was then freeze-thawed with liquid nitrogen three times to remove oxygen. After removing any oxygen from the reaction flask the Pd(PPh.3) ₄ and Cul was added and the reaction mixture was stirred overnight at room temperature. The reaction was worked up with aqueous saturated NH ₄ C1 (3 mL) and extracted with Et ₂ 0 (3 x 3 mL). The solvent was evaporated and the mixture was purified on silica gel using

EtOAc-hexanes (3:97) as the eluent to afford compound 27 as a clear oil (202 mg, 92%). NMR (500 MHz, CDCls) δ 6.54 (dd, J= 15.5, 10.9 Hz, 1H), 6.25 (dd, J= 15.3, 10.9 Hz, 1H), 6.04 (dd, J= 15.7, 5.4 Hz, 1H), 5.86 (dd, J= 15.3, 5.7 Hz, 1H), 5.73 - 5.55 (m, 2H), 5.51 - 5.40 (m, 1H), 5.40 - 5.26 (m, 1H), 4.58 (td, J = 6.0, 1.6 Hz, 1H), 4.33 (q, J = 5.5 Hz, 1H), 4.24 - 4.11 (m, 1H), 3.67 (s, 3H), 2.61 - 2.35 (m, 4H), 2.23 (dp, J= 20.8, 7.0 Hz, 2H), 2.11 - 1.95 (m, 4H), 0.95 (t, J= 7.6 Hz, 3H), 0.90 (s, 18H), 0.89 (s, 9H), 0.13 (s, 3H), 0.10 (s, 3H), 0.08 (s, 3H), 0.05 (s, 3H), 0.03 (s, 3H). NMR (500 MHz, CDCls) δ 173.92, 145.23, 141.21, 138.25, 133.99, 129.05, 124.28, 110.89, 109.15, 92.68, 87.10, 83.92, 80.71, 72.75, 71.83, 62.49, 51.71, 36.07, 33.67, 29.77, 29.75, 29.62, 26.00, 25.95, 25.94, 20.89, 18.41, 18.39, 14.34, -4.35, -4.41, -4.46, -4.65, -4.65, -4.92. (4S, 11R, 27S Trihydroxydocosa-(7¾ 9E, 15E, 2SZ tetraene-5,13-diynoic acid (3). To a solution of compound 27 (10 mg, 0.014 mmol) in THF (2 mL) was added 1.0 M solution of TBAF (0.084 mL, 0.084 mmol) at 0°C and stirred for 2 h. The reaction was quenched with water (3 mL) and extracted with Et ₂ 0 (5 x 3 mL), rinsed with brine, dried over MgS0 ₄ and filtered. The solvent was the concentrated and freshly prepared CH2N2 was added to convert any acid to the ester. The solvent was completely removed in vacuo and the compound was purified on silica gel using MeOH-CH2Cl2 (3%) as the eluent to afford an ester/lactone mixture (3:1 ratio see HPLC and NMR data attached). The product was then suspended in a H20-MeOH mixture (1:1, 1 mL) and NaOH (1 mg, 2.5 x lO ² mmol) was added. After 3 h the reaction mixture was dried and purified via C-18 reversed Phase HPLC using H2O-MeOH mixture (41%) to afford compound 3 as colorless oil (1.5 mg, 27%). Ή NMR (400 MHz, CD3OD) δ 6.59 (dd, J= 15.5, 10.8 Hz, 1H), 6.36 (dd, J= 15.4, 10.9 Hz, 1H), 6.03 (dd, J = 15.8, 6.1 Hz, 1H), 5.89 (dd, J= 15.1, 6.1 Hz, 1H), 5.68 (t, J= 14.9 Hz, 2H), 5.55 - 5.45 (m, 1H), 5.42 - 5.33 (m, 1H), 4.52 (dd, J= 6.7, 5.0 Hz, lH), 4.30 - 4.21 (m, 1H), 4.08 (q, J= 5.9 Hz, 1H), 2.57 - 2.48 (m, 1H), 2.39 (td, J = 15.6, 7.5 Hz, 3H), 2.28 (q, J= 7.5 Hz, 2H), 2.06 (dq, J= 12.3, 6.8, 6.0 Hz, 2H), 1.97 (q, J = 7.2 Hz, 2H), 0.98 (t, J= 7.5 Hz, 3H). NMR (400 MHz, CD3OD) δ 180.57, 144.28, 139.62, 136.98, 133.44, 123.68, 110.86, 109.50, 92.69, 85.94, 82.80, 79.96, 71.34, 70.10, 61.98, 61.94, 34.44, 34.20, 33.56, 27.53, 20.22, 13.08;

HRMS (ESI) m/z calcd for C22H27O5 : [M ]: 371.1864 found: 371.1881.

(4S, 11R, IZSJ-Trihydroxydocosa-^Z, 7E, 9E, 13Z, 15E, 2£Z hexaenoic acid, (1). A flame-dried flask was charged with a freshly prepared Zn/Cu/Ag amalgam (300 mg, excess) and suspended in H2O-MeOH mixture (1:1, 1 mL). To the reaction slurry was added compound 3 (1.5 mg, 3.8 x 10 ^s mmol) and stirred for 13 h while monitoring. The reaction was filtered dried and purified via HPLC at H2O-MeOH mixture (41%) to afford a mixture of the ester and gamma-butyrolactone. This was hydroiyzed in H2O-MeOH mixture using 3 eq of NaOH. Upon completion the mixture was dried and purified via HPLC to afford the compound (1) (0.33 mg, 22%). Ή NMR (600 MHz, CD ₃ OD) δ 6.60 (dd, J= 14.6, 11.5 Hz, 1H), 6.52 (dd, J= 15.4, 11.1 Hz, 1H), 6.34 (dd, J= 14.9, 10.8 Hz, 1H), 6.24 (dd, J= 14.6, 10.7 Hz, lH), 6.13 - 6.02 (m, 2H), 5.75 (dd, J = 15.2, 6.7 Hz, 1H), 5.70 (dd, J = 15.4, 6.5 Hz, 1H), 5.55 - 5.31 (m, 4H), 4.92 (s, 24H), 4.23 - 4.08 (m, 2H), 2.45 (q, J= 7.9 Hz, 2H), 2.38 - 2.22 (m, 4H), 2.07 (q, J= 7.3 Hz, 2H), 1.88 (dd, J = 14.1, 7.2 Hz, 1H), 1.82 - 1.71 (m, 1H), 0.98 (t, J = 7.5 Hz, 3H). i3C NMR (600 MHz, CD ₃ OD) δ 182.46, 137.54, 137.49, 135.48, 134.75, 134.58, 131.75, 131.04, 130.31, 129.15, 128.17, 126.58, 125.52, 73.20, 73.07, 68.86, 36.68, 36.23, 35.43, 35.28, 21.68, 14.54. ; HRMS (ESI) m/z calcd for C22H31O5 : [M-]: 375.2177 found: 375.2182.

(i?)-l-(i-Butyldiphenylsilyloxy)hept-4-yn-2-ol (30). This compound was prepared from protected S-Glycidol similarly to its enantiomer, compound 16. Ή NMR (400 MHz CDC1 ₃ ) δ 3.79 - 3.70 (m, 1H), 3.70 (dd, J= 9.8, 4.2 Hz, 1H), 3.60 (dd, J= 9.8, 5.8 Hz, 1H), 2.46 (s, 1H), 2.42 - 2.32 (m, 2H), 2.16 (dt, J= 7.5, 2.4 Hz, 2H), 1.11 (t, J= 7.5 Hz, 3H), 0.90 (s, 8H), 0.08 (s, 6H). NMR (400 MHz CDCls) δ 84.11, 75.17, 70.59, 65.77, 25.98, 23.51, 18.42, 14.29, 12.51, - 5.28, -5.31.

(3R, IE, 5 3-(i-Butyldimethylsilyloxy)-l-iodoocta-l,5-diene (31). This compound was prepared similarly to its enantiomer, compound 5. Ή NMR (400 MHz, CDCls) δ 6.53 (dd, J= 14.3, 5.7 Hz, 1H), 6.21 (dd, J= 14.3, 1.3 Hz, 1H), 5.54 - 5.41 (m, 1H), 5.38 - 5.25 (m, lH), 4.14 - 4.04 (m, 1H), 2.29 - 2.19 (m, 2H), 2.09 - 1.96 (m, 2H), 0.89 (s, 9H), 0.05 (s, 3H), 0.03 (s, 3H). C NMR (400 MHz, CDCls) δ 148.87, 134.40, 123.73, 77.48, 77.16, 76.84, 75.88, 75.18, 35.67, 25.95, 20.89, 18.37, 14.39, -4.49, -4.69.

Methyl (4S, 11R, 17R, 7E, 9E, 15E, 2P iris-(t-butyldimethylsnyloxy)-docosa- 7,9,15,19-tetraen-5,13-diynoate (32). This compound was prepared from alkyne 4 and vinyl iodide 31, similarly to its 17S-epimer, compound 27. Ή NMR (400 MHz, CDCls) δ 6.54 (dd, J= 15.5, 10.8 Hz, 1H), 6.25 (dd, J= 15.3, 10.9 Hz, 1H), 6.04 (dd, J = .15.7, 5.4 Hz, lH), 5.86 (dd, J = 15.0, 5.7 Hz, lH), 5.70 - 5.54 (m, 2H), 5.52 - 5.40 (m, 1H), 5.37 - 5.26 (m, lH), 4.58 (td, J= 6.1, 1.9 Hz, lH), 4.33 (q, J= 6.1 Hz, 1H), 4.15 (q, J= 6.3 Hz, 1H), 3.67 (s, 3H), 2.55 - 2.37 (m, 4H), 2.30 - 2.18 (m, 2H), 2.09 - 1.94 (m, 4H), 0.95 (t, 3H), 0.90 (s, 18H), 0.89 (s, 9H), 0.13 (s, 3H), 0.11 (s, 3H), 0.08 (s, 3H), 0.07 (s, 3H), 0.05 (s, 3H), 0.04 (s, 3H). we NMR (400 MHz, CDC1 ₃ ) δ 173.94, 145.23, 141.22, 138.26, 133.99, 129.06, 124.29, 110.90, 109.16, 92.69, 87.11, 83.93, 80.71, 72.76, 71.84, 62.49, 51.71, 36.08, 33.67, 29.87, 29.75, 29.62, 26.01, 25.96, 25.94, 20.89, 18.41, 18.39, 14.28, -4.35, -4.41, -4.46, -4.65, -4.65, -4.92.

(4S, 11R, iZR Trihydroxydocosa-(5Z, 7E, 9E, 13Z, 15E, 19Z)- exaenoic acid, (2). This compound was prepared from compound 32 similarly to its 17iS- epimer (1). The removal of the silyl protective groups led to a similar mixture of hydrolysis products, which were similarly converted to 2. ^X H NMR (600 MHz, CDsOD) δ 6.59 (dd, J= 14.7, 11.5 Hz, 1H), 6.52 (dd, J= 15.2, 11.2 Hz, 1H), 6.33 (dd, J = 15.1, 10.7 Hz, 1H), 6.24 (dd, J= 14.7, 10.9 Hz, 1H), 6.08 (td, J = 11.1, 4.3 Hz, 2H), 5.75 (dd, J = 15.1, 6.6 Hz, 1H), 5.70 (dd, J= 15.1, 6.6 Hz, 1H), 5.48 (td, J = 11.1, 5.8 Hz, 2H), 5.45 - 5.37 (m, 2H), 4.61 (q, J= 7.2 Hz, lH), 4.19 - 4.16 (m, 1H), 4.14 (q, J= 6.7 Hz, 1H), 2.52 - 2.38 (m, 2H), 2.30 (dq, J = 29.2, 7.3 Hz, 4H), 2.08 (dt, J= 14.4, 7.1 Hz, 2H), 1.87 (dt, J= 15.0, 7.3 Hz, 1H), 1.77 (dt, J= 13.7, 6.7 Hz, 1H), 0.98 (t, J= 7.5 Hz, 3H). NMR (600 MHz, CDsOD) δ 182.45, 137.54, 137.49, 135.48, 134.75, 134.58, 131.76, 131.04, 130.31, 129.15, 128.17, 126.58, 125.52, 73.20, 73.07, 68.86, 49.51, 49.34, 49.17, 49.00, 48.83, 48.66, 48.49, 36.68, 36.23, 35.43, 35.28, 21.68, 14.54; HRMS (ESI) m/z calcd for C22H31O5 : [M-]: 375.2177 found: 375.2185.

All references cited herein are incorporated herein by reference in their entirety.