WO2007110705A2 - Macrolide as inhibitors of mhc class ii - Google Patents

Abstract

The present invention provides macrolide compounds, and pharmaceutically acceptable salts thereof, that inhibit MHC class II expression and that can be used as immunosuppressive agents in the treatment prior to, during and/or after organ or tissue transplantation, as well as in the treatment of immune-related disorders and/or cancer and cancer-related disorders.

Description

MACROLIDE COMPOSITIONS AS THERAPEUTIC AGENT

Field of the Invention

This invention relates generally to methods of using compositions including a 12- membered ring macrolide compound as therapeutics in the treatment of immune-related disorders and cancer.

Background of the Invention

The immune system is highly complex and tightly regulated, with many alternative pathways capable of compensating deficiencies in other parts of the system. There are however occasions when the immune response becomes a cause of disease or other undesirable conditions if activated. Such diseases or undesirable conditions are, for example, autoimmune diseases, graft rejection after transplantation, allergy to innocuous antigens, psoriasis, chronic inflammatory diseases such as atherosclerosis, and inflammation in general. In these cases and others involving inappropriate or undesired immune response, there is a clinical need for immunosuppression.

In addition, the treatment of cancer has thus far proved problematic. While "cancers" share many characteristics in common, each particular cancer has its own specific characteristics. Genetics and environmental factors have a complex interplay in severity and prognosis of treatment. Thus, treatment must be carefully tailored. Certain pharmaceutical treatments have proven useful for one form of cancer, but not others. Other treatments such as radiation, while partially useful for a range of cancers, do not typically result in a complete cure. Indeed, given the severity of many cancers and the mortality rate, a drug can be deemed successful if it improves quality of life, e.g., by delaying growth of tumors, or prolongs life— without actually curing the condition. Thus, in many circumstances, an individual is treated with a compound or combination of treatments that can eliminate 90-95% of the malignant cells, but the remaining cells can regrow and metastasize, ultimately resulting in death.

Accordingly, there exists a need for compositions that can be used in the treatment of immune-related diseases and/or disorders, as well as compositions that are useful in the treatment of cancers and other related disorders. Summary of the Invention

The present invention is directed to compounds and pharmaceutically acceptable salts thereof including a macrolide compound that inhibits MHC class II expression and that can be used as an immunosuppressive agent in the treatment prior to, during and/or after organ or tissue transplantation, as well as in the treatment of immune-related disorders and/or cancer and cancer-related disorders. The macrolide compounds of the present invention include a twelve-membered ring structure such as the twelve-membered ring structure shown below in formula A for the macrolide compound N831 (A).

The compounds of the present invention have been modified at the R]₇ position of the macrolide ring. This application is related to the application U.S.S.N. 60/721,513, filed September 28, 2005. The compounds of this related application have been modified at the R₇ position of the macrolide ring.

The macrolide compounds of the invention include a twelve-membered ring. More particularly, the present invention includes compounds of Formula I:

or a pharmaceutically acceptable salt and/or an individual diastereomer thereof, wherein: Z is O, NH, S, or absent, such that when Z is absent the compound is represented by formula (F)

R_d is -NH₂ or azide.

R-₃ is hydrogen, silyl, C₁-C₆ allcyl or substituted Ci-C₆ alkyl, or -C(O)R_S.

Silyl is R^aR^bR^cSi-, further wherein R^a, R^b, and R^c are the same as or different from each other and each represents methyl, ethyl, i-propyl, t-butyl, or phenyl.

R₅ is methyl, -NH₂, Ci-C₆ alkylamino, CrC₆ dialkylamino, or heterocycle, each of which may be substituted.

R₄ is hydrogen.

R₅ and R_5a are each, independently, hydrogen or hydroxyl, or taken together form a carbonyl, or taken together, R₄ and any one of R₅ or Rs_a form a double bond.

R₆ is hydrogen, hydroxyl, Ci-C₆ alkyl or substituted C₁-C₆ alkyl.

R₇ is hydrogen or -C(O)R_t. R_t is methyl, -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted.

R₁₆ and Ri _6a are the same or different from each other and each represents: methyl, hydrogen, or hydroxyl.

Rn is:

(A) -OC(O)R_17b;

(B) sulfonate or substituted sulfonate;

(C) hydroxyl;

(D) silyloxy; or

Ri_7a is hydrogen, or taken together Rn and Ri _7a form a carbonyl. R_17b is -NR_jR_k, -OR_1n, or -R_n. R_j and R_k are the same as or different from eacli other and each represents: hydrogen; C₂, C₃, C₄, C₅, or C₆ alkyl; C₃, C₄, C₅, or C₆ alkenyl; C₂, C3, C₄, C5, or C₆ acyl; unsaturated C₃, C₄, C₅, C₆, C₇, or C₈ acyl; C₅, C₆, C₇, or C₈ aryl; heteroaryl; benzyl; Cj, C₂, C₃, C₄, C₅, or C₆ alkylsulfonyl; benzenesulfonyl; -(CH₂)_x-heteroaryl; C₃, C₄, C5, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkenyl; -(CH₂)_y-heterocycle; -(CH₂)_n-NH₂; or methyl, each of which may be substituted, x is 1, 2, or 3. y is O, 1, 2, or 3. n is 0, 1, 2, or 3.

Alternatively, -NR_jR_k is bound together to represent a ring, wherein the ring is a non- aromatic heterocyclic ring containing at least one nitrogen atom, wherein any of the atoms in the ring may be substituted.

R_m is C₂, C₃, C₄, C₅, or C₆ alkyl; C₃, C₄, C₅, or C₆ alkenyl; C₅, C₆, C7, or Cg aryl; heteroaryl; , benzyl; -(CH₂)_s-heteroaryl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkenyl; -(CH₂)_u-heterocycle, -(CH₂)_t-NH₂, or methyl, each of which may be substituted, s is 1, 2, or 3. t is 1, 2, or 3. u is O, 1, 2, or 3.

R_n is C₅, C₆, C₇, or C₈ aryl; heteroaryl; C₂, C₃, C₄, C₅, or C₆ alkyl; -(CH₂)_g-heterocycle; - (CH₂)i-C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; -(CH₂)_h- NH₂; -(CH₂)_j-heteroaryl; or methyl, each of which may be substituted. g is O, I, 2, or 3. h is 1, 2, or 3. j is O, 1, 2, or 3. i is 0, 1, 2, or 3.

R₁₈ is hydroxyl, halogen, or taken together any one of Rn or Ri _7a and R₁₈ are connected to form a 3, 4, 5, or 6-membered ring containing at least one oxygen atom. Ri₉ is hydroxyl, halogen, or -C(O)R₁₁, or taken together Rj₈ and R₁₉ are connected to form a 3, 4, 5, or 6-membered ring containing at least one oxygen atom or taken together Ri₈ and Ri ₉ form a double bond.

R₁₁ is methyl, aryloxy, -NH₂, Ci-C₆ alkoxy, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted. R₂i and R₂ u are the same or different from each other and each represents: hydrogen, hydroxy, C₁-C_O alkyl, Ci-C₆ alkoxy, or -OC(O)R_V, or taken together R_2] and R₂i_aform a carbonyl. R_v is methyl, aryloxy, -NH₂, Ci-C₆ alkoxy, CrC₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each which may be substituted. R₂₂ is hydrogen or Ci-C₆ alkyl.

In one embodiment, if R₇ is -COR_t, wherein R_t is -NH₂, CpC₆ alkylamino, or Ci-C₆ dialkylamino, then any one of R]₇ or Ri _7a is not hydroxyl when the other is hydrogen. In another embodiment, when the above compound is represented by Formula IA:

(IA) wherein, taken together R₄ and any one of R₅ or R_5a form a double bond and any one of R_2I or R₂ia is hydroxyl, then the other R₂i or R₂i_a is not hydrogen. When R₄, R5, and Rs₃ are each independently hydrogen and any one of R₂i or R₂i_a is methoxy or hydroxyl, then the other R₂I or R₂i_a is not hydrogen.

The present invention includes a method of treating an immune-related disorder comprising administering to a patient in need of such treatment a composition comprising a earlier and a non-toxic therapeutically effective amount of a macrolide compound of the present invention.

One aspect of the invention includes a method of treating an immune-related disorder which is an autoimmune disease selected from a connective tissue disease, a neuromuscular disease, an endocrine disease, a gastrointestinal disease, an autoimmune skin disease, a vasculitis syndrome, a hematologic autoimmune disease, and uveitis. In another embodiment of the invention, the autoimmune disease is selected from systemic lupus erythematosus (SLE), systemic sclerosis (scleroderma), Sjogren's syndrome, multiple sclerosis (MS), myasthenis gravis, Guillain-Barre syndrome, Hashimoto's thyroiditis, Graves' disease, insulin-dependent (Type 1) diabetes, inflammatory bowel disease, Crohn's disease, ulcerative colitis, and psoriatic arthritis. Another aspect of the invention includes a method of treating an immune-related disorder which is an inflammatory disorder selected from Alzheimer's disease, asthma, atopic allergy, allergy, bronchial asthma, diabetic retinopathy, atopic dermatitis (also called eczema), glomerulonephritis, graft vs. host disease, hemolytic anemias, sepsis, stroke, vasculitis, and ventilator induced lung injury.

Another embodiment of the invention includes a method of alleviating a symptom associated with an immune-related disorder comprising administering to a patient in need of such treatment a composition comprising a carrier and a non-toxic, therapeutically effective amount of a macrolide compound of the present invention.

One embodiment includes a method of alleviating a symptom associated with an immune-related disorder which is an autoimmune disease selected from a connective tissue disease, a neuromuscular disease, an endocrine disease, a gastrointestinal disease, an autoimmune skin disease, a vasculitis syndrome, a hematologic autoimmune disease, and uveitis.

Another embodiment includes a method of alleviating a symptom associated with an autoimmune disease which is selected from systemic lupus erythematosus (SLE), systemic sclerosis (scleroderma), Sjogren's syndrome, multiple sclerosis (MS), myasthenis gravis, Guillain-Barre syndrome, Hashimoto's thyroiditis, Graves' disease, insulin-dependent (Type 1) diabetes, inflammatory bowel disease, Crohn's disease, ulcerative colitis, and psoriatic arthritis.

Another embodiment includes a method of alleviating a symptom associated with an immune-related disorder which is an inflammatory disorder selected from Alzheimer's disease, asthma, atopic allergy, allergy, bronchial asthma, diabetic retinopathy, eczema, glomerulonephritis, graft vs. host disease, hemolytic anemias, sepsis, stroke, vasculitis, and ventilator induced lung injury.

In one embodiment, the macrolide used to treat an immune-related disorder is a racemic mixture of a macrolide compound of the present invention. In another embodiment, the macrolide used to treat an immune-related disorder is an enantiomerically pure form of a macrolide compound of the present invention.

In one embodiment, the macrolide used to treat an immune-related disorder is administered in combination with a second agent used to treat an immune-related disorder. In another embodiment, a second agent is in combination with the macrolide compound to treat an immune-related disorder. The second agent is selected from methotrexate, cyclosporin A, tacrolimus, corticosteroids, statins, interferon beta, nonsteroidal antiinflammatory drugs (NSAIDs), and the disease-modifying anti-rheumatic drugs (DMARDs). In a further embodiment, the second agent is cyclosporin A, further wherein the cyclosporin A is cyclosporin microemulsion.

Another aspect of the invention includes a method of suppressing an immune response associated with organ or tissue transplantation comprising administering to a patient in need of such treatment a composition comprising a carrier and a non-toxic, therapeutically effective amount of a macrolide compound of the present invention.

In one embodiment, the macrolide is administered in combination with a second agent used to suppress an immune response associated with organ or tissue transplantation. In a further embodiment, the second agent used to suppress an immune response associated with organ or tissue transplantation is selected from methotrexate, cyclosporin A, cyclosporin microemulsion, tacrolimus, corticosteroids and statins. In another embodiment, the second agent is cyclosporin A, further wherein the cyclosporin A is cyclosporin microemulsion.

In another embodiment, the macrolide administered in combination with a second agent used to suppress an immune response is an enantiomerically pure form of a macrolide compound of the present invention.

In another embodiment, the macrolide administered in combination with a second agent used to suppress an immune response is administered to said patient at a time selected from prior to said organ or tissue transplantation, during said organ or tissue transplantation, after said organ or tissue transplantation, and combinations thereof.

Another aspect of the invention includes a method of inhibiting tumor growth comprising administering to a patient in need of such treatment a composition comprising a carrier and a non-toxic therapeutically effective amount of a compound, wherein the compound is a macrolide compound of the present invention. In another embodiment, the macrolide is administered in combination with a second agent used to inhibit tumor growth.

( Detailed Description of the Invention

The details of at least one embodiments of the invention are set forth in the accompanying description below. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the methods and materials of the present invention are now described. Other features, objects, and advantages of the invention will be apparent from the description. In the specification, the singular forms also include the plural unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In the case of conflict, the present specification will control.

The present invention provides compositions that are used in the treatment of cancer and cancer-related disorders, in the treatment of immune-related disorders, as well as in the treatment prior to, during and/or after organ or tissue transplantation.

The compositions of the invention include a compound that modulates and/or regulates an immune response. This invention includes a macrolide compound. Macrolides are a group of antibiotics, produced for example by various strains of Streptomyces, that have a complex macrocyclic structure. These macrocyclic compounds are formed by chain extension and cyclized into a large, typically 12-membered ring containing a nitrogen, oxygen, or sulfur atom. Often, macrolides are glycosylated. This invention also includes ring-opened macrolide compounds as represented by the structure below:

More particularly, the present invention includes compounds of Formula I:

or a pharmaceutically acceptable salt and/or an individual diastereomer thereof. In one aspect of the invention Z is O, NH, S, or absent, such that when Z is absent the compound is represented by formula (F)

R_d is -NH₂ or azide. R₃ is hydrogen, silyl, Ci-C₆ alkyl or substituted Ci-C₆ alkyl, or - C(O)R₈. Silyl is R^aR^bR°Si- and R^a, R^b, and R^c are the same as or different from each other and each represents methyl, ethyl, i-propyl, t-butyl, or phenyl. R₃ is methyl, -NH₂, Ci-C₆ alkylamino, CpC₆ dialkylamino, or heterocycle, each of which may be substituted. R₄ is hydrogen. R₅ and R_5a are each, independently, hydrogen or hydroxyl, or taken together R₅ and R_5aform a carbonyl. In another aspect, taken together, R₄ and any one of R5 or Rs_a form a double bond.

R₆ is hydrogen, hydroxyl, Ci-C₆ alkyl or substituted Cj-C₆ alkyl.

R₇ is hydrogen or -C(O)R_t. R_t is methyl, -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted. R]₆ and Ri _6a are the same or different from each other and each represents: methyl, hydrogen, or hydroxyl. Rn is -OC(O)Ri₇b, sulfonate or substituted sulfonate, hydroxyl, or silyloxy. R]_7a is hydrogen or taken together Ri₇ and Ri _7a form a carbonyl.

R_17b is -NRjR_k, -OR_m, or -R_n.

R_j and R^ are the same as or different from each other and each represents: hydrogen; C₂, C₃, C₄, C₅, or C₆ alkyl; C₃, C₄, C₅, or C₆ alkenyl; C₂, C₃, C₄, C₅, or Cc acyl; unsaturated C₃, C₄, C₅, C₆, C₇, or C₈ acyl; C₅, C₆, C₇, or C₈ aryl; heteroaryl; benzyl; C₁, C₂, C₃, C₄, C₅, or C₆ alkylsulfonyl; benzenesulfonyl; -(CH₂)_x-heteroaryl; C₃, C₄, C₅, Ce, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkenyl; -(CH₂)_y-heterocycle; -(CH₂)_H-NH₂; or methyl, each of which may be substituted. x is 1, 2, or 3. y is O, 1, 2, or 3. n is 0, 1, 2, or 3.

Alternatively, -NR_jR_k is bound together to represent a ring, wherein the ring is a non-aromatic heterocyclic ring containing at least one nitrogen atom. Any of the atoms in the ring may be substituted.

R₁₁₁ is C₂, C₃, C₄, C₅, or C₆ alkyl; C₃, C₄, C₅, or C₆ alkenyl; C₅, C₆, C₇, or C₈ aryl; heteroaryl; benzyl; -(CH₂)_s-heteroaryl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkenyl; -(CH₂)_u~heterocycle, -(CH₂)_t-NH₂, or methyl, each of which may be substituted. s is 1, 2, or 3. t is 1, 2, or 3. u is 0, 1, 2, or 3.

R_n is C₅, C₆, C₇, or C₈ aryl; heteroaryl; C₂, C₃, C₄, C₅, or C₆ alkyl; -(CH₂)_g- heterocycle; -(CHa)₁-C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; -(CH₂)_h-NH₂; -(CH₂)_j-heteroaryl; or methyl, each of which may be substituted. g is 0, 1, 2, or 3. h is 1, 2, or 3. j is O, 1, 2, or 3. i is 0, 1, 2, or 3.

Ri₈ is hydroxyl, halogen, or taken together any one of Ri₇ or Ri_7a and Ri₈ are connected to form a 3, 4, 5, or 6-membered ring containing at least one oxygen atom. R₁₉ is hydroxyl, halogen, or -C(O)R₁₁, or taken together Ri₈ and R19 are connected to form a 3, 4, 5, or 6-membered ring containing at least one oxygen atom. R_u is methyl, aryloxy, -NH₂, Ci- C₆ alkoxy, Ci-Cg alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted, or taken together Rig and R₁₉ form a double bond.

R_2I and R_{2 la} are the same or different from each other and each represents: hydrogen, hydroxy, C₁-C₆ alkyl, Ci-C₆ alkoxy, or -OC(O)R_V, or taken together R₂i and R₂i_a form a carbonyl. R_v is methyl, aryloxy, -NH₂, Ci-C₆ alkoxy, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each which may be substituted. R₂₂ is hydrogen or Ci-C₆ alkyl. In one embodiment, if R₇ is -COR_t, wherein R_t is -NH₂, Ci-C₆ alkylamino, or Ci-C₆ dialkylamino, then any one of Rn or Ri _7a is not hydroxyl when the other is hydrogen. In another embodiment, when the above compound is represented by Formula IA:

(IA) and taken together R₄ and any one of R₅ or R_5a form a double bond and any one of R₂₁ or R₂i_a is hydroxyl, then the other R₂₁ or R₂j_a is not hydrogen. When R₄, R5, and R_5a are each independently hydrogen and any one of R₂₁ or R₂ i_a is methoxy or hydroxyl, then the other R₂i or R₂i_a is not hydrogen.

One embodiment of the present invention is a compound according to Formula II:

(II) or a pharmaceutically acceptable salt or individual diastereomer. Z is O, NH, or S. R₃ is hydrogen, silyl, Ci-C₆ alkyl or substituted Ci-C₆ alkyl, or -C(O)R₈. Silyl is R^aR^bR^cSi-, and R^a, R^b, and R^c are the same as or different from each other and each represents methyl, ethyl, i-propyl, t-butyl, or phenyl. R_s is methyl, -NH₂, C₁-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted.

R₆ is hydrogen, hydroxyl, Ci-C₆ alkyl or substituted Ci-C₆ alkyl. R₇ is hydrogen or - C(O)R_t. Ri is methyl, -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted.

Ri_7b is -NR_jR_k, -OR_m, or -R_n.

R_j and R_k are the same as or different from each other and each represents: hydrogen; C₂, C₃, C₄, C₅, or C₆ alkyl; C₃, C₄, C₅, or C₆ alkenyl; C₂, C₃, C₄, C₅, or C₆ acyl; unsaturated C₃, C₄, C₅, C₆, C₇, or Cs acyl; C₅, C₆, C₇, or C₈ aryl; heteroaryl; benzyl; Ci, C₂, C₃, C₄, C₅, or C₆ alkylsulfonyl; benzenesulfonyl; -(CH₂)_x-heteroaryl; C₃, C₄, C₅, C₆, C₇, or Cg cycloalkyl; C₃, C₄, C₅, C₆, C₇, or Cg cycloalkenyl; -(CH₂)_y-heterocycle; -(CH₂)_n-NH₂; or methyl, each of which may be substituted. x is 1, 2, or 3. y is 0, 1, 2, or 3. n is 0, 1, 2, or 3.

Alternatively, -NR_jR_k is bound together to represent a ring, wherein the ring is a non-aromatic heterocyclic ring containing at least one nitrogen atom. Any of the atoms in the ring may be substituted.

R_ra is C₂, C₃, C₄, C₅, or C₆ alkyl; C₃, C₄, C₅, or C₆ alkenyl; C₅, C₆, C₇, or C₈ aryl; heteroaryl; benzyl; ~(CH₂)_s-heteroaryl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkenyl; -(CH₂)_u-heterocycle, -(CH₂)_t-NH₂, or methyl, each of which may be substituted. s is 1, 2, or 3. t is 1, 2, or 3. u is 0, 1, 2, or 3.

R₁₁ is C₅, C₆, C₇, or C₈ aryl; heteroaryl; C₂, C₃, C₄, C₅, or C₆ alkyl; -(CH₂)_g- heterocycle; -(CH₂VC₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; -(CH₂)_h-NH₂; -(CH₂)_j-heteroaryl; or methyl, each of which may be substituted. g is 0, 1, 2, or 3. h is 1, 2, or 3. j is 0, 1, 2, or 3. i is O, 1, 2, or 3. In one embodiment, Z is O. In another embodiment, Z is NH. In another emobodiment, Z is S.

In one embodiment, R₆ is hydrogen. In another embodiment, R₇ is -C(O)R_t. In a further embodiment, R_t is methyl. In another embodiment, R_t is -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle. In another embodiment, R₃ is hydrogen. In a further embodiment, R₆ is hydrogen, R₇ is -C(O)CH₃, and R₃ is hydrogen.

In another embodiment, R₃ is -C(O)R_S and R_s is Ci-C₆ alkyl. In one embodiment, R_s is methyl. In another embodiment, R₇ is hydrogen.

In one embodiment, R_17b is -NRjR_k. In another embodiment, Rj and Rk are the same as or different from each other and each represents: hydrogen; C₂-C₆ alkyl; C₃-C₆ alkenyl; C_2-C₆ acyl; unsaturated C_3-Cs acyl; Cs-Cs aryl; heteroaryl; benzyl; Ci-C₆ alkylsulfonyl; benzenesulfonyl; or methyl, each of which may be substituted.

In a further embodiment, when R₁₇b is -NRjR_k, Rj and R_k are the same as or different from each other and each represents: -(CH₂)_xheteroaryl; C₃-C₈ cycloalkyl; C₃-C₈ cycloalkenyl; heterocycle; -(CH₂)_y-heterocycle; -(CH₂)_T1-NH₂; or hydrogen.

Alternatively, when Ri _7b is -NR_jR_k, -NR_jR_k is bound together to represent a 3, 4, 5, 6, 7, or 8-membered ring, wherein the ring is a non-aromatic heterocyclic ring containing at least one nitrogen atom, wherein any of the atoms in the ring may be substituted. In one embodiment, the ring is a 3-membered ring. In another embodiment, the ring is a 4- membered ring. In another embodiment, the ring is a 5-membered ring. In another embodiment, the ring is a 6-membered ring. In another embodiment, the ring is a 7- membered ring. In another embodiment, the ring is an 8-membered ring.

In one embodiment, when R_17b is -NR_jR_k bound together to represent a 3, 4, 5, 6, 7, or 8-membered ring, the ring contains at least two nitrogen atoms.

In another embodiment, when R_17b is bound together to represent a 3, 4, 5, 6, 7, or 8- membered ring, the ring is pyrrolidine, piperidine, azepane, tetrahydropyridine, tetrahydropyrimidine, morpholine, piperazine, homopiperazine, or azocane, each of which may be substituted.

In one embodiment, when R₁ ^ is -NRjR_kbound together to represent a ring, the ring is pyrrolidine. In a further embodiment, pyrrolidine is substituted with hydroxyl or Ci-C₆ alkyl. In another embodiment, pyrrolidine is substituted with methylhydroxyl. In another embodiment, when Ri_7b is -NRjR_k bound together to represent a ring, the ring is piperidine. In a further embodiment, the piperidine ring is substituted with hydroxyl or pyridine.

In another embodiment, when Ri _7b is -NR_jR_k bound together to represent a ring, the ring is 1,2,3,6-tetrahydropyridine or 1,2,5,6-tetrahydropyiϊdine. In a further embodiment, the 1,2,3,6-tetrahydropyridine or 1,2,5,6-tetrahydropyridine ring is substituted with pyridine.

In another embodiment, when Ri_7b is -NR_jR_k bound together to represent a ring, the ring is 2,4,5,6-tetrahydropyrimidine.

In another embodiment, when Ri _?b is -NR_jR_k bound together to represent a ring, the ring is morpholine.

In another embodiment, when Rπ_b is -NR_jR_k bound together to represent a ring, the ring is piperazine. In one embodiment, the piperazine ring is not substituted. In a further embodiment, the piperazine ring is substituted with one Of C₃-C₈ cycloalkyl, Ci-C₆ alkyl, heteroaryl, -C(O)R₀, Cs-C₈ aryl, or hydroxyl. In another embodiment, piperazine is substituted with pyridine, phenol, pyrimidine, phenyl, or cyclohexyl. In another embodiment, piperazine is substituted with -C(O)R_C, wherein R_c is Ci-Cβ alkyl. In one embodiment, R₀ is ethyl. In another embodiment, piperazine is substituted with Ci-Ce alkyl. In one embodiment, piperazine is substituted with methyl. In another embodiment, piperazine is substituted with ethyl. In a further embodiment, ethyl is substituted with hydroxyl, -NH₂, Ci-Cβ alkylamino, or Ci-C₆ dialkylamino.

In another embodiment, when Rπ_b is -NR_jR_k bound together to represent a ring, the ring is azepane. In another embodiment, when Ri ₇b is -NRjR_k bound together to represent a ring, the ring is homopiperazine.

In one embodiment, when Ri_7b is -NR_jRk, any one of R,- or R_k is selected from - (CH₂)_x-heteroaryl; C₃-C₈ cycloalkyl; -(CH₂)_y-heterocycle, or -(CH₂)_n-NH₂ and the other R_j or R_k is selected from hydrogen or methyl. In further embodiment, the other of Rj or R_k is hydrogen. In another embodiment, the other of Rj or R_k is methyl.

In one embodiment, when Ri _7b is -NR_jRk, any one of R_j or R_k is selected from - (CH₂)_x-heteroaryl. In one embodiment, x is 1. In another embodiment, x is 2. In another embodiment, x is 3. In one embodiment, heteroaryl is pyridine.

In one embodiment, when Rπb is -NRjRk, any one of R_j or R_k is C₃-C₈ cycloalkyl. In another embodiment, C₃-C₈ cycloalkyl is cyclohexyl. In one embodiment, when Rπ_b is -NRjR_k, any one of Rj or Rk is -(CH₂)_y-heterocycle and the other of R_j or R_k is hydrogen. In another embodiment, heterocycle is a 6-membered ring. In one embodiment, y is 0. In another embodiment, y is 1. In another embodiment, y is 2. In another embodiment, y is 3. In one embodiment, heterocycle is selected from pyrrolidine, piperidine, morpholine, pyridine, or piperazine, each of which may be substituted. In one embodiment, heterocycle is pyrrolidine. In another embodiment, heterocycle is pyridine. In another embodiment, heterocycle is piperidine. In another embodiment, heterocycle is morpholine. In another embodiment, heterocycle is piperazine. In a further embodiment, piperazine is substituted with methyl.

In one embodiment, when Rπ_b is -NR_jR_k, any one of Rj or R_k is -(CH₂)_n-NH₂. In another embodiment, the other of R_j or R_k is hydrogen. In another embodiment, the other of R_j or R_k is methyl. In one embodiment, n is 1. In another embodiment, n is 2. In another embodiment, n is 3. In another embodiment, when Rπb is -NR_jR_k and any one of Rj or R_k is -(CH₂)_I1-NH_2, -NH₂ is substituted with Ci-C₆alkyl. In one embodiment, -NH₂ is substituted to form a Ci-C₆alkylamino. In one embodiment, -NH₂ is substituted to form a Ci- Cedialkylamino. In another embodiment, -NH₂ is substituted with methyl to form

_

In one embodiment, when Rπb is -NR_jR_k, R_j and Rk are both Ci-C₆ alkyl. In another embodiment, R_j and R_k are both methyl. In another embodiment, any one of R_j or R_k is methyl and the other is ethyl. In a further embodiment, ethyl is substituted with hydroxyl. In one embodiment, when Rπ_b is -NR_jR_k, R_j and R_k are both hydrogen.

In another embodiment, Rπ_b is -NR_jR_k and R₇ is -O(C)R_t. In one embodiment, when R_πb is -NR_jR_k and R₇ is -O(C)R_t,R_t is methyl, -NH₂, C₁-C₆ alkylamino, C₁-C₆ dialkylamino, or heterocycle, each of which may be substituted. In another embodiment, Rπ_b is -NR_jR_k and R₇ is -0(C)Rt and Rt is -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, each of which may be substituted. In one embodiment, when Rπb is -NRjR_k and R₇ is - 0(C)R_1, R_j and R_k are the same as or different from each other and each represents: hydrogen; C₂, C₃, C₄, C₅, or C₆ alkyl; C₃, C₄, C₅, or C₆ alkenyl; C₂, C₃, C₄, C₅, or C₆ acyl; unsaturated C₃, C₄, C₅, C₆, C₇, or Cs acyl; C₅, C₆, C₇, or C₈ aryl; heterparyl; benzyl; C₁, C₂, C₃, C₄, C₅, or C₆ alkylsulfonyl; benzenesulfonyl; -(CH₂)_x-heteroaryl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkenyl; -(CH₂)y-heterocycle; -(CH₂)_n-NH₂; or methyl, each of which may be substituted. x is 1, 2, or 3. y is O, 1, 2, or 3. n is O, I, 2, or 3.

Alternatively, -NR_jR_k is bound together to represent a ring, wherein the ring is a non-aromatic heterocyclic ring containing at least one nitrogen atom. Any of the atoms in the heterocycle ring may be substituted. In one embodiment, where Rj ₇b is -NRjRk and R₇ is -0(C)R_t, any one of R_j or R_k is -(CH₂)_y-heterocycle and the other is hydrogen. In a further embodiment, y is 2 and heterocycle is morpholine. In another embodiment, where Ri _7b is - NRjR_k and R₇ is -0(C)Rt, R_t is Ci-C₆ alkylamino. In one embodiment, Ci-C₆ alkylamino is ethylamine substituted with morpholine.

In another embodiment, Rπ_b is -OR_m. In one embodiment, R_n, is C₂-C₆ alkyl; C₃-C₆ alkenyl; Cs-C₈ aryl; heteroaryl; benzyl; -(CH₂)_s-heteroaryl; C₃-C₈ cycloalkyl; C₃-C₈ cycloalkenyl; -(CEbXrheterocycle, -(CH₂X-NH₂, or methyl, each of which may be substituted. In one embodiment, s is 1, 2, or 3. In another embodiment, t is 1, 2, or 3. In another embodiment, u is 0, 1, 2, or 3. In a further embodiment, R_m is C₂-C₆ alkyl; C₃-C₆ alkenyl; Cs-C₈ aryl; heteroaryl; or methyl, each of which may be substituted. In another embodiment, R_m is -(CH₂)_s-aryl; -(CH₂)_s-heteroaryl; C₃-C₈ cycloalkyl; C₃-C₈ cycloalkenyl; - (CH₂)_u-heterocycle; or -(CH^^-ammo, each of which may be substituted.

In one embodiment, when Ri_7b is -OR_m, R_m is C₅-C₈ aryl. In a further embodiment, C₅-C₈ aryl is phenyl. In another embodiment, phenyl is substituted with -NO₂.

In another embodiment, Rπ_b is R_n. In one embodiment, R_n is C₅-C₈ aryl; heteroaryl; C₂-C₆ alkyl; -(CH₂)_g-heterocycle; -(CH₂)I-C₃-C₈ cycloalkyl; C₃-C₈ cycloalkyl; -(CH₂)_h- NH₂; -(CH₂)_j-heteroaryl; or methyl, each of which may be substituted. In one embodiment, g is 0, 1, 2, or 3. In another embodiment, h is 1, 2, or 3. In another embodiment, j is 0, 1, 2, or 3. In another embodiment, i is 0, 1, 2, or 3.

In one embodiment, when Rπb is R_n, R_n is C₅-C₈ aryl or heteroaryl, each of which may be substituted. In one embodiment, R_n is phenyl or pyridine.

In another embodiment, when Rπ_b is R_n, R_n is Ci-C₆ alkyl; -(CH₂)_g-heterocycle; - (CH₂)_J-C₃-C₈ cycloalkyl; C₃-C₈ cycloalkyl; -(CH₂)_h-NH₂; or -(CH₂)_j-heteroaryl, each of which may be substituted. In another embodiment, when Rπb is R_n, R_n is selected from C₁-C₆ alkyl, -(CH₂)_n- NH₂, or C₃-C₈ cycloalkyl, each which may be substituted. In one embodiment, R_n is methyl or cyclopropyl.

In another embodiment, when Rπ_b is R_n> R_n is -(CH₂)_n-NH₂. In one embodiment, h is l.

Another embodiment of the present invention is a compound according to Formula III:

or a pharmaceutically acceptable salt or individual diastereomer. Z is O, NH, or S. R₃ is hydrogen, silyl, or -C(O)R₃. Silyl is R^aR^bR^cSi-, and R^a, R^b, and R^c are the same as or different from each other and each represents methyl, ethyl, i-propyl, t-butyl, or phenyl, R₃ is methyl, -NH₂, alkylamino, dialkylamino, or heterocycle, each which may be substituted. R₇ is hydrogen or -C(O)CH₃.

Ri₆ and R_16a are the same or different from each other and each represents: methyl, hydrogen, or hydroxyl. R₁₇ is hydroxyl, silyloxy; or taken together Ri₇ and Ri_7a form a carbonyl; or Ri_7a is hydrogen. Ri₈ is hydroxyl or halogen, or taken together Ri₇ and R₁₈ are connected to form a 3, 4, 5, or 6-membered ring containing at least one oxygen atom.

Ri₉ is hydroxyl, halogen, or -C(O)R₁₁; or taken together Ri₈ and R₁₉ are connected to form a 3, 4, 5, or 6-membered ring containing at least one oxygen atom. R_u is methyl, aryloxy, -NH₂, C₁-Cn alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted.

R_2I and R₂i_a are the same or different from each other and each represents: hydrogen, hydroxyl, Ci-C₆ alkyl, Ci-C₆ alkoxy, or -OC(O)R_V, or taken together R₂i and R₂i_a form a carbonyl. R_v is methyl, aryloxy, -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted.

R₂₂ is hydrogen or Ci-C₆ alkyl. When the above compound is represented by Formula IIIA:

(IIIA), if any one of R₂i or R_2Ia is methoxy or hydroxyl, then the other R2₁ or R₂i_a is not hydrogen.

In one embodiment, Z is O. In another embodiment, Z is NH. In another embodiment, Z is S.

In one embodiment, R₃ is -C(O)CH₃. In another embodiment, R₃ is hydrogen. In another embodiment R₃ is -C(O)R_S. In one embodiment, R_s is methyl, -NH₂, alkylamino, dialkylamino, or heterocycle, each which may be substituted. In another embodiment, R_s is dialkylamino. In another embodiment, R_s is dimethylamine.

In one embodiment, R₇ is -C(O)CH₃. In another embodiment, R₇ is hydrogen. In one embodiment, Ri ₈ is a halogen. In another embodiment, Rig is bromide, chloride, or iodide. In one embodiment, R₁₉ is halogen. In another embodiment, R₁9 is bromide, chloride, or iodide.

In one embodiment, Rig and R₁₉ are connected to form an epoxide ring.

In one embodiment, R₁₉ is -C(O)R_U. In a further embodiment, R₁₁ is aryloxy, -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle. In one embodiment, R₂i is C₁-

Cβ alkoxy. In one embodiment, R₂i is methoxy. In another embodiment, any one of R₂i or R_2Ia is -OC(O)Rv and the other is hydrogen. In another embodiment, any one of R₂i or R_2]a is -OC(O)Ry and the other is hydrogen, and R_vis -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted. In one embodiment, R_v is Ci-C₆ alkylamino. In another embodiment, R_v is ethylamine. In a further embodiment, ethylamine is substituted with pyrrolidine.

In one embodiment, any one of Rj₇ or Ri _7a is hydroxyl and the other is hydrogen. In another embodiment, Ri₇ and Ri₈ are connected to form a 3, 4, 5, or 6-membered ring containing at least one oxygen atom. In one embodiment, R₁₇ and Ri₈ are connected to form a cyclic carbonate ring. In another embodiment, Rn and R]₈ are connected to form an epoxide ring.

In another embodiment, R₁₉ is -C(O)R₁₁. In one embodiment, R₁₄ is aryloxy, -NH₂, alkylamino, dialkylamino, or heterocycle. In one embodiment, R₁₉ is hydroxyl.

In another embodiment, Ri₇ and R]₈ are each hydroxyl. In another embodiment, Ri₇ and R]₉ are each hydroxyl. In another embodiment, taken together, Rj₇ and Ri_7a form a carbonyl. In another embodiment, taken together, R₂i and R₂i_a form a carbonyl. In another embodiment, any one of R₂i or Rπ_a is methyl and the other is hydroxyl. In one embodiment, R₇ is -C(O)CH₃, R₃ is hydrogen, and R₂1 is methoxy.

In another embodiment, R₂₂ is Ci-C₆ alkyl or hydrogen. In one embodiment, R₂₂ is hydrogen. In another embodiment, R₂₂ is Ci-C₆ alkyl. In a further embodiment, R₂₂ is methyl.

In another embodiment, Ri₆ and Ri_6a are different. In one embodiment, Ri₆ and Ri_6a are Ci-C₆ alkyl and hydroxyl. In another embodiment, Ri₆ and Ri_6a are methyl and hydroxyl.

Another embodiment of the present invention is a compound according to Formula

IV:

(IV) or a pharmaceutically acceptable salt or individual diastereomer thereof, wherein Z is O, NH, or S. R₃ is hydrogen, Ci-C₆ alkyl or substituted Ci-C₆ allcyl. R₄ is hydrogen. R5 and R_5a are each independently hydrogen or hydroxyl, or taken together form a carbonyl, or taken together, R₄ and any one of R₅ or R₅₃ form a double bond.

R₆ is hydrogen, hydroxyl, Ci-C₆ alkyl or substituted Ci-C₆ alkyl. Taken together Ri₈ and Ri₉ form a double bond or are connected to form a 3, 4, 5, or 6-membered ring containing at least one oxygen atom. R₂i and R₂i_a are the same or different from each other and each represents: hydrogen, hydroxyl, or Ci-C₆ alkoxy, or taken together R₂₁ and R₂ u form a carbonyl.

When the above compound is represented by Formula IVA:

(IVA) if any of one R₂] or R₂i_a is hydroxyl, then the other R₂i or R₂i_a is not hydrogen.

In one embodiment, Z is O. In another embodiment, Z is NH. In another embodiment, Z is S.

In one embodiment, R₃ is hydrogen. In another embodiment, R₆ is hydroxyl. In another embodiment, R₆ is hydrogen. In another embodiment, R₃ is hydrogen and R₆ is hydroxyl.

In one embodiment, Ris and R₁₉ are connected to form an epoxide ring.

In another embodiment, any one of R₂₁ or R₂i_a is hydroxyl. In a further embodiment, the other of R₂i or R₂i_a is hydrogen.

In another embodiment, any one of R₅ or Rs_a taken together with R₄ forms a double bond. In another embodiment, taken together, R₅ and R_5a form a carbonyl. In another embodiment, taken together, R]₈ and R₁₉ form a double bond. In another embodiment, any one of R₅ or Rs_a and R₄ are each hydroxyl. In one embodiment, R5 and Rs_aform a double bond and taken together R₂] and R₂] _a form a carbonyl.

Some representative compounds of the invention are shown in Table 1. Table 1: Macrolide Compounds

Definitions

For convenience, certain terms used in the specification, examples and appended claims are collected here.

"Alkyl" includes saturated aliphatic groups, including straight-chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl), branched-chain alkyl groups (e.g., isopropyl, tert-butyl, isobutyl). "Alkyl" further includes alkyl groups that have oxygen, nitrogen, or sulfur atoms replacing one or more hydrocarbon backbone carbon atoms. Alkyl further includes alkyl groups that have unsaturation e.g.,

. In certain embodiments, a straight chain or branched alkyl has six or fewer carbon atoms in its backbone {e.g., methyl, Ci-C₆ for straight chain, C₃-C₆ for branched chain). In another embodiment, a straight chain or branched alkyl has four or fewer carbon atoms in its backbone.

The teπn "alkyl" also includes both "un substituted" and "substituted alkyls", the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbon of the hydrocarbon backbone. Such substitutents can include, for example, alkyl, alkenyl, alkynyl, hydroxyl, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, ester alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, cyano, -NH₂, alkylamino, dialkylamino, arylamino, diarlylamino, alkylarylamino, acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl, and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, tliiocarboxylate, sulfates, alkylsulfϊnyl, sulfonato, sulfamoyl, aminesulfoxide, sulfonamide, nitro, -CF₃, halogen, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety. "Alkyl" also includes the side chains of natural and unnatural amino acids. The term "Ci-C₆ alkyl" includes alkyl groups with 1, 2, 3, 4, 5, or 6 carbon atoms. The term

"C₂-C_O alkyl" includes alkyl groups with 2, 3, 4, 5, or 6 carbon atoms. Aryl includes groups with aromaticity, including 5, 6, 7, or 8-membered "unconjugated", or single-ring aromatic groups that may include from one to four heteroatoms, as well as "conjugated", or multicyclic systems with at least one aromatic ring. Examples of aryl groups include phenyl, pyrrole, furan, thiophene, thiazole, isothiazole, imidazole, triazole, tetrazole, pyrazole, oxazole, isoxazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like. Furthermore, the term "aryl" includes multi-cyclic groups, e.g., tricyclic, bicyclic, e.g., naphthalene, benzoxazole, benzodioxazole, benzothizole, benzoimidazole, benzothiophene, methylenedioxyphenyl, quinoline, isoquinoline, napthridine, indole, benzofuran, purine, benzofuran, deazapureine, or indolizine. Those aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles", "heterocycles", "heterocyclyls", "heteroaryls" or "heteroaromatics" e.g., pyridine, pyrazole, pyrimidine, furan, isoxazole, imidazole[2,l,b]thiazole, triazole, pyrazine, benzothiophene, imidazole, or thiophene.

The aromatic ring can be substituted at one or more ring positions with such substituents as described above, as for example, halogen, hydroxyl, alkyl, alkoxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid, alkylcarbonyl, alkylaminocarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl, allcylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, carboxyalkyl, cyano, -NH₂, alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino, acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety. Aryl groups can also be fused or bridged with alicyclic or heterocyclic rings, which are not aromatic so as to form a multicyclic system (e.g., tetralin, methylenedioxyphenyl) .

"Alkenyl" includes unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double bond. For example, the term "alkenyl" includes straight-chain alkenyl groups (e.g., ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl), branched-chain alkenyl groups, cycloalkenyl (e.g., alicyclic) groups (e.g., cyclopropenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl or alkenyl substituted cycloalkenyl groups, and cycloalkyl or cycloalkenyl substituted alkenyl groups. The term "alkenyl" further includes alkenyl groups, which include oxygen, nitrogen, or sulfur replacing one or more hydrocarbon backbone carbons. In certain embodiments, a straight chain or branched chain alkenyl group has six or fewer carbon atoms in its backbone (e.g., C₂-C₆ for straight chain, C₃-C₆ for branched chain). Likewise, cycloalkenyl groups may have from three to eight carbon atoms in their ring structure, and more preferably have five or six carbons in the ring structure. The term "C₂-C₆" includes alkenyl groups containing 2, 3, 4, 5, or 6 carbon atoms. The term "C₃-C₆" includes alkenyl groups containing 3, 4, 5, or 6 carbon atoms.

The term "alkenyl" also includes both "unsubstituted alkenyls" and "substituted alkenyls", the latter of which refers to alkenyl moieties having substituents replacing a hydrogen on one or more hydrocarbon backbone carbon atoms. Such substituents can include, for example, alkyl groups, alkenyl groups, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl, hydroxyl, nitro, trifluoromethyl, cyano, azido, phenyl, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

"Alkynyl" includes unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one triple bond. For example, "alkynyl" includes straight chain alkynyl groups (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl), branched chain alkynyl groups, and cycloalkyl or cycloallcenyl substituted alkynyl groups. The term "alkynyl" further includes alkynyl groups having oxygen, nitrogen, sulfur or phosphorous atoms replacing one or more hydrocarbon backbone carbons. In certain embodiments, a straight chain or branched chain alkynyl group has six or fewer carbon atoms in its backbone (e.g., C₂-C₆ for straight chain, C₃-C₆ for branched chain). The term "C₂-C₆ alkynyl" includes alkynyl groups containing 2, 3, 4, 5, or 6 carbon atoms.

The term "alkynyl" also includes both "unsubstituted alkynyls" and "substituted alkynyls", the latter of which refers to alkynyl moieties having substituents replacing a hydrogen on one or more hydrocarbon backbone carbon atoms. Such substituents can include, for example, alkyl groups, alkenyl groups, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylamiiiocarbonyl, dialkylaminocarbonyl, allcylthiocarbonyl, alkoxyl, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfϊnyl, sulfonato, sulfamoyl, sulfonamido, nilxo, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

"Acyl" includes compounds and moieties that contain the acyl radical (CH₃CO-) or a carbonyl group. "Substituted acyl" includes acyl groups where one or more of the hydrogen atoms are replaced by for example, alkyl groups, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, cyano, -NH₂, alkylamino, dialkylamino, arylamino, diarylamino, and allcylarylamino, acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

The term "alkoxycarbonyl" or "ester" includes compounds and moieties that contain an oxygen atom covalently linked to a carbonyl group (-OC(O)-). Examples of ester groups include -OC(O)CH₃, -OC(O)CH₂NH₂.

The term "alkoxy" or "alkoxyl" includes substituted and unsubstituted alkyl, alkenyl, and alkynyl groups covalently linked to an oxygen atom. Examples of alkoxy groups (or alkoxyl radicals) include methoxy, ethoxy, isopropyloxy, propoxy, butoxy, and pentoxy groups. Examples of substituted alkoxy groups include halogenated alkoxy groups. The alkoxy groups can be substituted with groups such as alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, ester, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, cyano, -NH₂, alkylamino, dialkylamino, aiylamino, diarylamino, and alkylarylamino, acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, siilfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl, liydroxyl, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties. Examples of halogen substituted allcoxy groups include, but are not limited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy, chloromethoxy, dichloromethoxy, and trichloromethoxy. The term "C₁-C₆ alkoxy" includes alkoxy groups which include 1, 2, 3, 4, 5, or 6 carbon atoms.

The term "cycloalkyl" includes saturated acyclic groups (e.g., cyclopropyl, cyclopentyl, cyclohexyl, cyclohexyl, cycloheptyl, cyclooctyl). Cycloalkyls have from three to eight carbon atoms in their ring structure. In certain embodiments, cycloalkyls have five or six carbon atoms in the ring structure. Cycloalkyls includes both "unsubstituted cycloalkyls" and "substituted cycloalkyls", the latter of which refers to replacing a hydrogen on one or more of the carbons in the ring structure. Such substituents can include, for example, alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, ester, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, cyano, -NH₂, alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino, acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety. The term "C₃-C₈ cycloalkyl" includes cycloalkyl groups with 3, 4, 5, 6, 7, or 8 carbon atoms.

The terms "heterocycle," "heterocyclyl," or "heterocyclic group" include closed ring structures, e.g., 3, 4, 5, 6, 7, 8, 9, or 10-, or 4, 5, 6, or 7-membered rings, which include one or more heteroatoms. "Heteroatom" includes atoms of any element other than carbon or hydrogen. Examples of heteroatoms include nitrogen, oxygen, or sulfur.

Heterocycle groups can be saturated or unsaturated and include pyrrolidine, pyrazine, pyrimidine, oxolane, 1,3-dioxolane, thiolane, tetrahydrofuran, tetrahydropyran, tetrahydropyridine, tetrahydropyrimidine, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams such as azetidinones and pyrrolidinones, sultams, and sultones.

Heterocyclic groups such as pyrrole and furan can have aromatic character. They include fused ring structures such as quinoline and isoquinoline. Other examples of heterocyclic groups include pyridine and purine. The heterocyclic ring can be substituted at one or more positions with such substituents as described above, as for example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid, alkylcarbonyl, alkoxycarbonyl, ester, aminocarbonyl, allcylthiocarbonyl, alkoxyl, cyano, -NH₂, alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino, acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, sulfonato, sulfamoyl, hydroxyl, nitro, trifluoromethyl, cyano, azido, heterocyclyl, or an aromatic or heteroaromatic moiety. Heterocyclic groups can also be substituted at one or more constituent atoms with, for example, a lower alkyl, a lower alkenyl, a lower alkoxy, a lower alkylthio, a lower alkylamino, a lower alkylcarboxyl, a nitro, a hydroxyl, -CF₃, or - CN, or the like.

The term "thioalkyl" includes compounds or moieties which contain an alkyl group connected with a sulfur atom. The thioalkyl groups can be substituted with groups such as alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, ester, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, cyano, -NH₂, alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino, acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfmyl, sulfonato, sulfamoyl, hydroxyl, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties.

The teπn "thiocarbonyl" or "thiocarboxy" includes compounds and moieties which contain a carbon connected with a double bond to a sulfur atom.

The term "hydroxy" or "hydroxyl" includes groups with an -OH or -O^".

The term "halogen" includes fluorine, bromine, chlorine, iodine, etc. The term "perhalogenated" generally refers to a moiety wherein all hydrogen atoms are replaced by halogen atoms.

The term "carbamoyl" includes compounds and moieties which include the following arrangement of atoms -NHC(O)O- or -OC(O)NH-. The term "carbonate" includes compounds and moieties which include the following arrangement of atoms -OC(O)O-.

The term sulfonate includes compounds and moieties which contain the sulfate ion

a sulfone group. "Substituted sulfonate" includes sulfonate groups where the hydrogen atom is replaced by for example, alkyl groups or aryl aryl groups. Examples of substituted sulfonate groups are methanesulfonate, trifluormethanesulfonate or p- toluenesulfonate .

A "pharmaceutically acceptable salt" or "salt" of one or more of the disclosed compounds is a product of the disclosed compound that contains an ionic bond, and is typically produced by reacting the disclosed compound with either an acid or a base, suitable for administering to a subject.

A "pharmaceutical composition" is a formulation containing one or more of the disclosed compounds in a form suitable for administration to a subject. In a preferred embodiment, the pharmaceutical composition is in bulk or in unit dosage form. The unit dosage form is any of a variety of forms, including, for example, a capsule, an IV bag, a tablet, a single pump on an aerosol inhaler, or a vial. The quantity of active ingredient (e.g., a formulation of the disclosed compound or salts thereof) in a unit dose of composition is an effective amount and is varied according to the particular treatment involved. One skilled in the art will appreciate that it is sometimes necessary to make routine variations to the dosage depending on the age and condition of the patient. The dosage will also depend on the route of administration. A variety of routes are contemplated, including oral, pulmonary, rectal, parenteral, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, intranasal, and the like. Dosage forms for the topical or transdermal administration of a compound of this invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. In a preferred embodiment, the active compound is mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that are required.

An "anionic group," as used herein, refers to a group that is negatively charged at physiological pH. Anionic groups include carboxylate, sulfate, sulfonate, sulfmate, sulfamate, tetrazolyl, phosphate, phosphonate, phosphinate, or phosphorothioate or functional equivalents thereof. "Functional equivalents" of anionic groups are intended to include bioisosteres, e.g., bioisosteres of a carboxylate group. Bioisosteres encompass both classical bioisosteric equivalents and non-classical bioisosteric equivalents. Classical and non-classical bioisosteres are known in the art (see, e.g., Silverman, R. B. The Organic Chemistry of Drug Design and Drug Action, Academic Press, Inc.: San Diego, Calif., 1992, pp.19-23). One example of an anionic group is a carboxylate.

The term "unstable functionality" refers to a substitution pattern that contains a labile linkage, e.g., a functionality or bond that is susceptible to hydrolysis or cleavage under physiological conditions (e.g., aqueous solutions in the neutral pH range). Examples of unstable functionalities include acetals and ketals.

The terms "crystal polymorphs" or "polymorphs" refer to the existence of more than one crystal form for a compound, salt or solvate thereof. Crystal polymorphs of the doxepin-analog compounds are prepared by crystallization under different conditions.

It will be noted that the structure of some of the macrolide compounds of the invention includes asymmetric carbon atoms and thus are capable of existing as an enantiomer. It is to be understood accordingly that the isomers arising from such asymmetry (e.g., all enantiomeric and diastereomeric forms of a compound, including racemates or racemic mixtures) are included within the scope of the invention, unless indicated otherwise. In some cases, there may be advantages (i.e., the compound may have greater efficacy), to using a particular enantiomer when compared to the other enantiomer or the racemate or racemic mixture in the methods of the instant invention and such advantages can be readily determined by those skilled in the art. Such isomers can be obtained in substantially pure form by classical separation techniques and by stereochemically controlled synthesis. Furthermore, the structures and other compounds and moieties discussed in this application also include all tautomers thereof. Alkenes can include either the E- or Z-geometry, where appropriate.

Further, the structures and other compounds discussed in this application include all atropic isomers thereof. Atropic isomers are a type of stereoisomer in which the atoms of two isomers are arranged differently in space. Atropic isomers owe their existence to a restricted rotation caused by hindrance of rotation of large groups about a central bond. Such atropic isomers typically exist as a mixture, however as a result of recent advances in chromatography techniques, it has been possible to separate mixtures of two atropic isomers in select cases. As used herein, the term "analog" refers to a chemical compound that is structurally similar to another but differs slightly in composition (as in the replacement of one atom by an atom of a different element or in the presence of a particular functional group, or the replacement of one functional group by another functional group). Thus, an analog is a compound that is similar or comparable in function and appearance, but not in structure or origin to the reference compound.

As defined herein, the term "derivative", e.g., in the term "macrolide derivatives", refers to compounds that have a common core structure, and are substituted with various groups as described herein. For example, all of the compounds represented by formula are macrolide derivatives, and have one of formula 1, 1', II, III, or IV as a common core.

The macrolide compositions of the invention are used as immunosuppression agents in organ or tissue transplantation. As used herein, "immunosuppression agent" refers to an agent whose action on the immune system leads to the immediate or delayed reduction of the activity of at least one pathway involved in an immune response, whether this response is naturally occurring or artificially triggered, whether this response takes place as part of the innate immune system, the adaptive immune system, or both. For example, the macrolide compositions of the invention regulate the expression of Major Histocompatibility Complex (MHC) class II genes. MHC class II molecules are directly involved in the activation of T lymphocytes and in the control of the immune response. MHC class II expression occurs on the surface of antigen-presenting cells (APCs). These cells are capable of presenting antigen to lymphocyte T-helpers, which control the development of an immune response. Thus, the expression of MHC class II molecules is the important to antigen presentation. Only a limited number of specialized cell types express MHC class II constitutively, numerous other cells become MHC class II positive upon stimulation, e.g., by a cytokine such as, for example, interferon gamma (IFN-γ).

These immunosuppressive macrolide compositions are administered to a subject prior to, during and/or after organ or tissue transplantation. For example, the macrolide compositions of the invention are used to treat or prevent rejection after organ or tissue transplantation.

In one embodiment, these immunosuppressive macrolide compositions are administered in combination with any of a variety of known anti-inflammatory and/or immunosuppressive compounds. Suitable anti-inflammatory and/or immunosuppressive compounds for use with the macrolide compounds of the invention include, but are not limited to, methotrexate, cyclosporine A (including, for example, cyclosporine microemulsion), tacrolimus, corticosteroids and statins.

In addition, the macrolide compositions of the invention are used to treat or alleviate a symptom associated with cancer and cancer-related disorders e.g. a cell proliferative disorder. As used herein, the term "cell proliferative disorder" refers to conditions in which the unregulated and/or abnormal growth of cells can lead to the development of an unwanted condition or disease, which can be cancerous or non-cancerous, for example a psoriatic condition. As used herein, the term "psoriatic condition" refers to disorders involving keratinocyte hyperproliferation, inflammatory cell infiltration, and cytokine alteration.

In one embodiment, the cell proliferation disorder is cancer. As used herein, the term "cancer" includes solid tumors, such as lung, pancreas, stomach, rectum, kidney, breast, cervical/uterine, testicular, colon, ovarian, prostate, head and neck, espophageal, malignant melanoma, non-melanoma skin cancers, as well as hematologic tumors and/or malignancies, such as childhood leukemia and lymphomas, multiple myeloma, Hodgkin's disease, lymphomas of lymphocytic and cutaneous origin, acute and chronic leukemia such as acute lymphoblastic, acute myelocytic or chronic myelocytic leukemia, plasma cell neoplasm, lymphoid neoplasm, non-malignant neoplasias, and cancers associated with AIDS.

In addition to psoriatic conditions, the types of proliferative diseases which may be treated using the compositions of the present invention are epidermic and dermoid cysts, lipomas, adenomas, capillary and cutaneous hemangiomas, lymphangiomas, nevi lesions, teratomas, nephromas, myofibromatosis, osteoplastic tumors, and other dysplastic masses and the like. In one embodiment, proliferative diseases include dysplasias and disorders of the like. The compositions include a pharmaceutically acceptable carrier and a compound according to formula I, I', II, III, IV, or pharmaceutically acceptable salts or hydrates thereof.

The macrolide compositions of the invention are also used to treat or alleviate a symptom associated with an immune-related disorder, such as, for example, an autoimmune disease or an inflammatory disorder. The compositions include a pharmaceutically acceptable carrier and a compound according to formula I, I', II, III, IV, or pharmaceutically acceptable salts or hydrates thereof. Autoimmune diseases include, for example, Acquired Immunodeficiency Syndrome (AIDS, which is a viral disease with an autoimmune component), alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease (AIED), autoimmune lymphoproliferative syndrome (ALPS), autoimmune thrombocytopenic purpura (ATP), Behcet's disease, cardiomyopathy, celiac sprue- dermatitis hepetiformis; chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy (CIPD), cicatricial pemphigoid, cold agglutinin disease, crest syndrome, Crohn's disease, Degos' disease, deπnatomyositis- juvenile, discoid lupus, essential mixed cryoglobulinemia, fibrόmyalgia-fibromyositis, Graves' disease, Guillain-Barre syndrome, Hashimoto's thyroiditis, idiopathic pulmonary fibrosis, idiopathic thrombocytopenia purpura (ITP), IgA nephropathy, insulin-dependent diabetes mellitus, juvenile chronic arthritis (Still's disease), juvenile rheumatoid arthritis, Meniere's disease, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, Cyclospori anemia, polyarteritis nodosa, polychondritis, polyglandular syndromes, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomena, Reiter's syndrome, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma (progressive systemic sclerosis (PSS), also known as systemic sclerosis (SS)), Sjogren's syndrome, stiff- man syndrome, systemic lupus erythematosus, Takayasu arteritis, temporal arteritis/giant cell arteritis, ulcerative colitis, uveitis, vitiligo and Wegener's granulomatosis. Inflammatory disorders, include, for example, chronic and acute inflammatory disorders. Examples of inflammatory disorders include Alzheimer's disease, asthma, atopic allergy, allergy, atherosclerosis, bronchial asthma, atopic dermatitis (also called eczema), glomerulonephritis, graft vs. host disease, hemolytic anemias, osteoarthritis, sepsis, stroke, transplantation of tissue and organs, vasculitis, diabetic retinopathy and ventilator induced lung injury. Atopic dermatitis is characterized by the distinctive phenomena of atopy and includes allergic contact dermatitis, irritant contact dermatitis, infantile seborrhoeic eczema, adult seborrhoeic eczema, varicose eczema, and discoid eczema.

In one embodiment, the macrolide compositions used to treat an immune-related disorder are administered in combination with any of a variety of known anti-inflammatory and/or immunosuppressive compounds. Suitable known compounds include, but are not limited to methotrexate, cyclosporine A (including, for example, cyclosporine microemulsion), tacrolimus, corticosteroids, statins, interferon beta, Remicade (Infliximab), Eribrel (Etanercept) and Humira (Adalimumab).

For example, in the treatment of rheumatoid arthritis, the macrolide compositions of the invention are co-administered with corticosteroids, methotrexate, cyclosporine A, statins, Remicade (Infliximab), Enbrel (Etanercept) and/or Humira (Adalimumab).

In the treatment of uveitis, the macrolide compositions are administered in conjunction with, e.g., corticosteroids, methotrexate, cyclosporine A, cyclophosphamide and/or statins. Likewise, patients afflicted with a disease such as Crohn's Disease or psoriasis are treated with a combination of a macrolide composition of the invention and Remicaid (Infliximab), and/or Humira (Adalimumab).

Patients with multiple sclerosis receive a combination of a macrolide composition of the invention in combination with, e.g., glatiramer acetate (Copaxone), interferon beta-la (Avonex), interferon beta-la (Rebif), interferon beta-lb (Betaseron or Betaferon), mitoxantrone (Novantrone), dexamethasone (Decadron), methylprednisolone (Depo- Medrol), prednisone (Deltasone) and/or statins.

The present invention also provides methods of treating or alleviating a symptom associated with an immune-related disorder or a symptom associated with rejection following organ transplantation. For example, the compositions of the invention are used to treat or alleviate a symptom of any of the autoimmune diseases and inflammatory disorders described herein.

Symptoms of an immune-related disorder include, for example, inflammation, fever, loss of appetite, weight loss, abdominal symptoms such as, for example, abdominal pain, diarrhea or constipation, joint pain or aches (arthralgia), fatigue, rash, anemia, extreme sensitivity to cold (Raynaud's phenomenon), muscle weakness, muscle fatigue, changes in skin or tissue tone, shortness of breath or other abnormal breathing patterns, chest pain or constriction of the chest muscles, abnormal heart rate (e.g., elevated or lowered), light sensitivity, blurry or otherwise abnormal vision, and reduced organ function.

As described above, the macrolide compositions of the invention are administered alone, or alternatively, in combination with another form of therapy, referred to herein as "combination therapy" (or "co-therapy"). Co-therapy includes the administration of a macrolide composition of the invention and known therapy for a given immune-related disorder, or set of immune-related disorders. For example, a macrolide composition administered as an immunosuppressive agent in the treatment and/or prevention of rejection following organ transplantation is "co-administered" with one or more known immunosuppressive agents, such as, for example cyclosporine A, tacrolimus, and corticosteroids. Macrolide compositions administered in the treatment of an autoimmune disease and/or inflammatory disorder is administered with any of a variety of known therapies. Such therapies include, for example, known immunosuppressive agents such as COX2 inhibitors, corticosteroids, statins, cannabinoids (and derivatives thereof), interferon beta, aspirin and other anti-inflammatory agents, inhibitors of tumor necrosis factor, and inhibitors of interleukin 1.

The beneficial effect of the combination can include, but is not limited to, pharmacokinetic or pharmacodynamic co-action resulting from the combination of therapeutic agents. Combinations of the compound of the present invention and the other active agents may be administered together in a single combination or separately. Where separate administration is employed, the administration of one element may be prior to, concurrent with, or subsequent to the administration of other agents. Administration of these therapeutic agents in combination typically is carried out over a defined time period (usually minutes, hours, days or weeks depending upon the combination selected). In one embodiment, "combination therapy" encompasses the administration of two or more of these therapeutic agents as part of separate monotherapy regimens that incidentally and arbitrarily result in the combinations of the present invention. In another embodiment, "combination therapy" is intended to embrace administration of these therapeutic agents in a sequential manner, that is, wherein each therapeutic agent is administered at a different time, as well as administration of these therapeutic agents, or at least two of the therapeutic agents, in a substantially simultaneous manner. Substantially simultaneous administration can be accomplished, for example, by administering to the subject a single capsule having a fixed ratio of each therapeutic agent or in multiple, single capsules for each of the therapeutic agents.

The therapeutic agents can be administered by the same route or by different routes. For example, a first therapeutic agent of the combination selected may be administered by intravenous injection while the other therapeutic agents of the combination may be administered orally. Alternatively, for example, all therapeutic agents may be administered orally or all therapeutic agents may be administered by intravenous injection. The sequence in which the therapeutic agents are administered is not narrowly critical. "Combination therapy" also can embrace the administration of the therapeutic agents as described above in further combination with other biologically active ingredients and non-drug therapies ( e.g., surgery or radiation treatment.) Where the combination therapy further comprises a non- drug treatment, the non-drug treatment may be conducted at any suitable time so long as a beneficial effect from the co-action of the combination of the therapeutic agents and non- drug treatment is achieved. For example, in appropriate cases, the beneficial effect is still achieved when the non-drug treatment is temporally removed from the administration of the therapeutic agents, perhaps by days or even weeks.

To evaluate whether a patient is benefiting from the administration of a macrolide, alone or in combination, one examines the patient's symptoms and/or immune response in a quantitative way, and compares the patient's symptoms and/or immune response before and after treatment with the macrolide compound. For example, a patient's symptom are determined by measuring a particular symptom, or set of symptoms, in a patient before and after treatment with a macrolide composition. For example, one measures and monitors symptoms such as fever, joint pain, muscle weakness using any of the standard measurement techniques known in the art. In a successful treatment, the patient status has improved (i.e., the measurement number has decreased, or the time to sustained progression has increased).

A compound that is administered in a pharmaceutical composition is mixed with a suitable carrier or excipient such that a therapeutically effective amount is present in the composition. The term "therapeutically effective amount" refers to an amount of the compound that is necessary to achieve a desired endpoint (e.g., decreasing symptoms associated with an immune-related disorder).

A variety of preparations can be used to formulate pharmaceutical compositions containing the macrolide compound, including solid, semi solid, liquid and gaseous forms. Techniques for formulation and administration are found, for example, in "Remington: The Science and Practice of Pharmacy, Twentieth Edition," Lippincott Williams & Wilkins, Philadelphia, PA. Tablets, capsules, pills, powders, granules, dragees, gels, slurries, ointments, solutions suppositories, injections, inhalants and aerosols are examples of such formulations. The formulations are administered in either a local or systemic manner or in a depot or sustained release fashion. Administration of the composition is performed in a variety of ways. Among others, oral, buccal, rectal, parenteral, intraperitoneal, intradermal, transdermal, intranasal, and intratracheal means can be used. The compositions and combination therapies of the invention are administered in combination with a variety of pharmaceutical excipients, including stabilizing agents, carriers and/or encapsulation formulations as described herein.

The preparation of pharmaceutical or pharmacological compositions will be known to those of skill in the art in light of the present disclosure. Typically, such compositions of the present invention are prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid prior to injection; as tablets or other solids for oral administration; as time release capsules; or in any other form currently used, including topical administration e.g., creams, lotions, mouthwashes, inhalants and the like.

For human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by FDA Office of Biologies standards.

Administration of compounds alone or in combination therapies are, e.g., intraperitoneal, ICV, intralesional, intraperitoneal, intramuscular,intravenous or subcutaneous injection, infusion, implant, inhalation spray, vaginal, rectal, sublingual, aerosol, topical, nasal, oral, ocular or otic delivery. The compounds can be administered on a regimen of 1 to 4 times per day. A particularly convenient frequency for the administration of the compounds of the invention is once or twice a day.

Upon formulation, therapeutics are administered in a manner compatible with the dosage formulation, and in such amount as is pharmacologically effective. The formulations are easily administered in a variety of dosage forms, such as the injectable solutions described, but drug release capsules and the like can also be employed. In this context, the quantity of active ingredient and volume of composition to be administered depends on the host animal to be treated. Precise amounts of active compound required for administration depend on the judgment of the practitioner and are peculiar to each individual.

A minimal volume of a composition required to disperse the active compounds is typically utilized. Suitable regimes for administration are also variable, but are typified by initially administering the compound and monitoring the results and then giving further controlled doses at further intervals. For example, for parenteral administration, a suitably buffered, and if necessary, isotonic aqueous solution is prepared and used for intravenous, intramuscular, subcutaneous or even intraperitoneal administration. One dosage is dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermolysis fluid or injected at the proposed site of infusion, (see for example, "Remington: The Science and Practice of Pharmacy, Twentieth Edition," Lippincott Williams & Wilkins, Philadelphia, PA).

A carrier can he a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and 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 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.

In some embodiments, compositions of the invention are formulated for parenteral administration, e.g., formulated for injection via the intravenous, intramuscular, subcutaneous, intralesional, or even intraperitoneal routes. The preparation of a composition that contains a compound or combination therapy of the invention, or an active component or ingredient will be known to those of skill in the art in light of the present disclosure. Typically, such compositions can be prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for using to prepare solutions or suspensions upon the addition of a liquid prior to injection can also be prepared; and the preparations can also be emulsified.

Suitable preservatives for use in solution include benzalkonium chloride, benzethonium chloride, chlorobutanol, thimerosal and the like. Suitable buffers include boric acid, sodium and potassium bicarbonate, sodium and potassium borates, sodium and potassium carbonate, sodium acetate, sodium biphosphate and the like, in amounts sufficient to maintain the pH at between about pH 6 and pH 8, and preferably, between about pH 7 and pH 7.5. Suitable tonicity agents are dextran 40, dextran 70, dextrose, glycerin, potassium chloride, propylene glycol, sodium chloride, and the like, such that the sodium chloride equivalent of the ophthalmic solution is in the range 0.9 plus or minus 0.2%. Suitable antioxidants and stabilizers include sodium bisulfite, sodium metabisulflte, sodium thiosulfϊte, thiourea and the like. Suitable wetting and clarifying agents include polysorbate 80, polysorbate 20, poloxamer 282 and tyloxapol. Suitable viscosity-increasing 14

agents include dextran 40, dextran 70, gelatin, glycerin, hydroxyethylcellulose, hydroxniethylpropylcellulose, lanolin, methylcellulose, petrolatum, polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, carboxymethylcellulose and the like.

The compounds of the invention can be formulated by dissolving, suspending or emulsifying in an aqueous or nonaqueous solvent. Vegetable (e.g., sesame oil, peanut oil) or similar oils, synthetic aliphatic acid glycerides, esters of higher aliphatic acids and propylene glycol are examples of nonaqueous solvents. Aqueous solutions such as Hank's solution, Ringer's solution or physiological saline buffer can also be used. In all cases the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.

Solutions of active compounds as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure.

The preparation of more, or highly, concentrated solutions for intramuscular injection is also contemplated. In this regard, the use of DMSO as solvent is preferred as this will result in extremely rapid penetration, delivering high concentrations of the active compound(s) or agent(s) to a small area.

Where the composition or combination therapy is given orally, it can be formulated alone or together through combination with conventional, non-toxic pharmaceutically acceptable carriers, adjuvants, and vehicles that are well known in the art. The carriers, adjuvants, and vehicles enable the compound to be formulated, for example, as a tablet, pill, troche, lozenge, hard or soft capsule, solution, aqueous or oily suspension, sustained release formulation, dispersible powder or granule, syrup, elixir, liquid or gel for oral ingestion by the patient. Oral use formulations can be obtained in a variety of ways, including mixing the compound with a solid excipient, optionally grinding the resulting mixture, adding suitable auxiliaries and processing the granule mixture. The following list includes examples of excipients that can be used in an oral formulation: sugars such as lactose, sucrose, mannitol or sorbitol; cellulose preparations such as maize starch, wheat starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidone (PVP), inert diluents, such as calcium carbonate, sodium carbonate, calcium phosphate or sodium phosphate; and lubricating agents, for example, stearic acid, or talc. Oral formulations include such normally employed excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate and the like. The tablets may be uncoated or coated by known techniques to delay disintegration and adsorporption in the gastrointestinal tract and thereby provide a sustained action over a period of time.

In certain defined embodiments, oral pharmaceutical compositions will comprise an inert diluent or assimilable edible carrier, or they may be enclosed in hard or soft shell gelatin capsule, or they may be compressed into tablets, or they may be incorporated directly with the food of the diet. For oral therapeutic administration, the active compounds may be incorporated with excipients and used in the form of ingestible tablets, buccal tables, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 0.1% of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 75% of the weight of the unit, or preferably between 25-60%. The amount of active compounds in such therapeutically useful compositions is such that a suitable dosage will be obtained. The tablets, troches, pills, capsules and the like may also contain the following: a binder, as gum tragacanth, acacia, cornstarch, or gelatin; excipients, such as dicalcium. phosphate; a disintegrating agent, such as corn starch, potato starch, alginic acid and the like; a lubricant, such as magnesium stearate; and a sweetening agent, such as sucrose, lactose or saccharin may be added or a flavoring agent, such as peppermint, oil of wintergreen, or cherry flavoring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier. Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance, tablets, pills, or capsules may be coated with shellac, sugar or both. A syrup of elixir may contain the active compounds sucrose as a sweetening agent methyl and propylparabensas preservatives, a dye and flavoring, such as cherry or orange flavor.

Oily suspensions may be formulated for oral delivery by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid parafϊin. The oily suspension may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents and flavoring agents such as those set out above may be added to provide a palatable oral preparation. The compositions may be preserved by the additiona of an anti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueous suspension for oral deliverby the addition of water provide the active ingredient in a mixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Additional exicipents, for example, sweeting, flavoring and coloring agents may also be present.

The composition of the invention may also be in the form of oil-in-water emulsions. The oily phase may be vegetable oil, for example, olive oil or arachis oil, or a mineral oil, for example, liquid paraffin or mixtures of these. Suitable emulsifying agents may be naturally occurring gums, for example gum acacia or gum tragacanth, naturally-occurring phosphatides, for example soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monoleate, and condensation products of the said partial esters with ethylene oxide. The emulsions may also contain sweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for example, glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative and flavoring and coloring agents. The compounds of the present invention can be delivered topically. 'Topical application", "applied topically", "topical administration", and "administered topically", are used interchangeably to mean the process of applying or spreading one or more compounds according to the present invention onto the surface of the skin or mucous membrane of a subject in need thereof. In one embodiment, compounds of the invention are incorporated into a topical preparation suitable for pharmaceutical applications. For topical use, creams, ointments, jellies, hydrogels, salves, sprays, foams, mousse, aerosols, emulsions, nanoemulsions, microemulsion, solutions or suspensions are employed. Topical applications may include mouthwashes and gargles. In another embodiment, the compound can be included in a transdermal patch of the matrix or reservoir type as are conventional in the art. When the compound is administered by a patch, the dose administration will be continuous rather than intermittent throughout the dosage regimen.

For topical administration, the compound of the invention may be combined with other optional suitable ingredients such as estrogen, Vitamin A, C, and E, alpha-hydroxy of alpha-keto acids such as pyruvic, lactic or glycolic acids, lanolin, vaseline, aloe vera, methyl or propyl paraben, pigments and the like. Suitable topically acceptable carriers include water, petroleum jelly (vaseline), mineral oil, vegetable oil, animal oil, organic and inorganic waxes, such as microcrystalline, paraffin and ozocerite wax, natural polymers, such as xanthanes, gelatin, cellulose, collagen, starch, or gum arabic, synthetic polymers, such as discussed below, alcohols, polyols, and the like. Excipients include solvents, surfactants, emollients, preservatives, colorants, fragrances and the like. In one embodiment, the carrier is a water miscible carrier composition that is substantially miscible in water. Such water miscible topical carrier composition can include those made with one or more appropriate ingredients set forth above but can also include sustained or delayed release carrier, including water containing, water dispersable or water soluble compositions, such as liposomes, microsponges, microspheres or microcapsules, aqueous base ointments, water-in-oil or oil-in-water emulsions, gels or the like.

The compounds of the present invention can also be in the form of an emulsion for topical administration. A stable emulsion is a mixture of two immiscible liquids, i.e. liquids that are not mutually soluble, but which can form a fluid in which very small droplets of one component are stably dispersed throught the other liquid, giving the mixture the appearance of a homogeneous fluid. Emulsions can include particulate materials and materials which are solid or solid-like at room temperature, but which will liquify at higher temperatures used during formulation of the emulsion. The presence of an emulsifier enhances the ability of one of the immiscible liquids to remain in a continuous form, while allowing the other immiscible liquid to remain in a dispersed droplet form. Thus, one function of an emulsifier is to provide a thickening or "'bodying" to an emulsion. Typically, emulsifiers are molecules with non-polar parts and polar parts that are able to reside at the interface of two immiscible liquids.

The term "emulsion" is used herein to identify oil-in-water (o/w) or water-in-oil (w/o) type dispersion formulations intended for applications to the skin, particularly lotions and creams providing cosmetic or therapeutic benefits. The emulsions may contain any number of desired "active" ingredients, including skin colorants, drug substances (such antiinflammatory agents, antibiotics, topical anesthetics, antimycotics, keratolytics, etc.), skin protectants or conditioners, humectants, ultraviolet radiation absorbers and the like, depending on the intended uses for the formulation. Techniques for forming o/w and w/o emulsions are well known in the art.

The compositions of the present invention can be delivered in an aerosol spray preparation from a pressurized pack, a nebulizer or from a dry powder inhaler. Suitable propellants that can be used in a nebulizer include, for example, dichlorodifluoro-methane, trichlorofluoromethane, dichlorotetrafluoroethane and carbon dioxide. The dosage can be determined by providing a valve to deliver a regulated amount of the compound in the case of a pressurized aerosol.Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as set out above. Preferably the compositions are administered by the oral or nasal respiratory route for local or systemic effect. Compositions in preferably sterile pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be breathed directly from the nebulizing device or the nebulizing device may be attached to a face mask, tent or intermittent positive pressure breathing machine. Solution, suspension or powder compositions may be administered, preferably orally or nasally, from devices which deliver the formulation in an appropriate manner.

Additional formulations suitable for other modes of administration include suppositories. These compositions can be prepared by mixing the drug with a suitable non- irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melot in the rectum to release the drug. For suppositories, traditional binders and carriers may include, for example, polyalkylene glycols or triglycerides; such suppositories may be formed from mixtures containing the active ingredient in the range of 0.5% to 10%, preferably l%-2%.

The subject treated by the methods of the invention is a mammal, more preferably a human. The following properties or applications of these methods will essentially be described for humans although they may also be applied to non-human mammals, e.g., apes, monkeys, dogs, mice, rats, horses, cattle, sheep, cats, etc. The invention therefore can also be tised in a veterinarian context.

A "pharmaceutically acceptable salt" or "salt" of the disclosed compound is a product of the disclosed compound that contains an ionic bond, and is typically produced by reacting the disclosed compound with either an acid or a base, suitable for administering to a subject.

A "pharmaceutical composition" is a formulation containing the disclosed compounds in a form suitable for administration to a subject. In one embodiment, the pharmaceutical composition is in bulk or in unit dosage form. The unit dosage form is any of a variety of forms, including, for example, a capsule, an IV bag, a tablet, a single pump on an aerosol inhaler, or a vial. The quantity of active ingredient (e.g. , a formulation of the disclosed compound or salts thereof) in a unit dose of composition is an effective amount and is varied according to the particular treatment involved. One skilled in the art will appreciate that it is sometimes necessary to make routine variations to the dosage depending on the age and condition of the patient. The dosage will also depend on the route of administration. A variety of routes are contemplated, including oral, pulmonary, rectal, parenteral, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, intranasal, and the like. Dosage forms for the topical or transdermal administration of a compound of this invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. For example, the active compound is mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that are required.

Pharmaceutical compositions of the present invention contain a therapeutically effective amount of the macrolide compound. The amount of the compound will depend on the patient being treated. The patient' s weight, severity of illness, manner of administration and judgment of the prescribing physician should be taken into account in deciding the proper amount. The determination of a therapeutically effective amount of a macrolide compound is well within the capabilities of one with skill in the art.

Although a therapeutically effective amount of a macrolide compound will vary according to the patient being treated, suitable doses will typically include between about 0.1 mg and 1000 mg of the compound per kg patient body weight per day which can be administered in single or multiple doses. In one embodiment, a dose contains between about 0.1 mg and 500 mg/kg per day of the compound. In another embodiment, a dose contains between about 0.1 mg and 250 mg/kg per day of the compound, from about 0.1 to 100 mg/kg per day. A suitable dosage level can be from about 0.1 to 50 mg/kg per day. Within this range the dosage maybe from 0.1 to about 0.5, from about 0.5 to about 5, or from about 5 to about 50 mg/kg per day.

It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy. In some cases, where it may be necessary to use dosages outside of the stated ranges to treat a patient, those cases will be apparent to the prescribing physician. Where it is necessary, a physician will also know how and when to interrupt, adjust or terminate treatment in conjunction with a response of a particular patient.

The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

Examples Example 1: Cytotoxicity of Macrolide Compounds

The cytotoxicity of each of the macrolide compounds of the invention was evaluated in the following cell types: HEPG2, ME67.8, HL60, PC3, Ovcar-3 and Colo205.

The cell lines were cultured according to the suppliers' protocols. For cytotoxicity measurements the cells were seeded in 96 or 384 well tissue culture treated microtitre plates overnight. Compounds were diluted in appropriate buffers and added to the cells the next day. Following a further 72 hour incubation, cell viability was determined using standard tetrazolium dyes (MTT and XTT from Sigma) or Cell Titer GIo (from Promega) according to the manufacturers' instructions and the absorbance or luminescence measured in a Tecan Ultra or PerkinElmer Microbeta microplate reader. Both methods gave very similar results (IC₅₀ values). The results of this evaluation are presented below in Table 2:

Table 2: MHCII activity and Cytotoxicity of Macrolide Compounds

Example 3: In Vivo Studies with N831 (A)

N831 (A) maximum tolerated dose (MTD)

The maximum tolerated dose of N831 (A) in Harlan nude mice is 10 mg/kg as established by the amount of weight loss observed. In the course of this study no fatalities and no adverse reactions were reported. At the 5 and 10 mg/kg doses, the mice lost around 20% of their body weight, all of which they had regained by the next week. Tumour growth delay xenograft study in mice

N831 (A) was evaluated for efficacy in athymic nude mice bearing xenografts of human PC3 prostate carcinoma.

Nude mice bearing PC3 xenografts were placed into groups often animals each. N831 (A) was administered daily intravenously for 5 days. The dose levels of N831 (A) were 2.5, 5.0 and 10 mg/kg. Response was assessed by tumor growth delay compared with untreated control using an endpoint of 1000 mm³ tumor volume, and by numbers of tumor regressions. All dose levels of N831 (A) were highly effective in delaying tumor growth of PC3 prostate carcinoma with time to endpoint values of 59.5-60.0 days. The tumor growth delay was highly significant when compared to control. A dose-response was evident in the number of regressions.

Example 4: Isolation of N831 (A) from T658

Microorganism and Fermentation

The Actinomycete strain T658 was isolated from marine sediments collected in waters off Singapore. The strain has been identified as a member of the genus Streptomyces owing to the presence of aerial mycelia and long, straight or flexuous spore chains. Currently species level identification is not available.

Strain T658 was subcultured on ISP4 agar (Difco) for 7 - 15 days at 28 ⁰C. As T658 is a marine derived organism the media was supplemented with 4% sea salts (Sigma). The subculture was used to inoculate 250 niL Erlenmeyer flasks each containing 50 mL of seed medium composed of 1.5% glucose (Sigma), 1.5% glycerol solution (Merck), 1.5% soy peptone (Oxoid), 4% sea salts and 0.1% CaCO₃ (BDH). The pH of the medium was adjusted to 7 prior to sterilization (autoclave 121 ⁰C for 20 min). The seed culture was incubated for 3 days at 28 ⁰C on a rotary shaker at 200 rpm. For fermentations using SP-207 resin (Mitsubishi Chemicals), the resin was removed from the fermentation broth using a combination of centrifugation (Sorvall RC-4), washing and decanting. The fermentation broth was first transferred to centrifuge containers and using the "quick run" mode the centrifuge was operated for 30 seconds (around 1300 rpm was obtained). This allowed the SP-207 beads to settle but most of the biomass remained suspended in the supernatant and could be easily poured off. Following this process the beads were washed with water and the remaining biomass was removed by decanting.

Isolation ofN831 (A), N1407, N1512, N1S13, N1S14, NlSlS, N1516, N1519 andN1551

The fermentation broth (48 L) was eluted through SP-207 resin and washed with MeOH. The MeOH eluent was added with H₂O (1:1) and partitioned 3 times with CH₂Cl₂. The active CH₂Cl₂ fraction (51 g) was chromatographed on Silica gel (700 g) with a CHCl₃ - MeOH gradient (100:0 → 5:1) to yield 10 fractions. Fractions 3 - 5 were combined and subjected to reverse phase preparative HPLC (isocratic; 18 mL/min; (acetonitrile + H₂O (30:70 over 20 min) follow by gradient elution;l 8 mL/min; (acetonitrile + H₂O (30:70 → 32:68 over 20 min, 32:68 → 35:65 over 25 min, 35:65 → 40:60 over 15 min, 40:60 → 50:50 over 10 min, 50:50 → 100:0 over 8 min.); Waters NovaPak C-18 radial cartridge column, 4O x 100 mm) to give the following compounds collected at the following retention time ("RT"): N831 (A) (1100 mg, RT 50 min), N1407 (2 mg, RT 50 min), N1512 (20 mg, RT 21.5 min), N1513 (3 mg, RT 90 min), N1514 (3 mg, RT 34 min), N1515 (3 mg, RT 70 min), N1516 (3 mg, RT 75 min), N1519 (1 mg, RT 45 min) and N1551 (6 mg, RT 88.5 min).

Isolation ofN1523 andNlόll

T658 was subjected to UV mutagenesis and antibiotic resistance to streptomycin to provide a mutant strain, which was found to produce some additional active compounds. The mutant strain was cultured following the standard protocol to provide a fermentation broth (4 L), which was eluted through SP-207 and washed with MeOH. The MeOH eluent was added with H₂O (1:1) and partitioned 3 times with CH₂Cl₂. The active CH₂Cl₂ fraction (2.5 g) was subjected to reverse phase preparative HPLC (gradient elution;18 mL/min; (acetonitrile + H₂O (10:90 → 15:85 over 10 min, 15:85 → 20:80 over 10 min, 20:80 → 25:75 over 20 min, 25:75 → 30:70 over 20 min, 30:70 → 35:65 over 20 min, 35:65 → 40:60 over 20 min, 40:60 → 50:50 over 20 min); Waters NovaPak C-18 radial cartridge column, 40 x 100 mm) to give N831 (A) (80 mg, RT 87 min), Nl 512 (20 mg, RT 73 min), N1523 (65 mg, RT 70 min) and N1611 (6 mg, RT 77 min). N831 (A)

¹H NMR (500 MHz, CDCl₃) δ 0.88 (3H, d, J = 7.1 Hz), 0.88 (3H, d, J = 6.9 Hz), 0.90 (3H₃ t, J = 7.5 Hz), 1.15 (3H, d, J = 6.9 Hz), 1.22 (3H, s), 1.29 (IH, m), 1.39 (IH, m), 1.41 (IH, m), 1.47 (IH, m), 1.55 (IH, m), 1.65 (IH, m), 1.71 (IH, m), 1.75 (3H, s), 2.01 (IH, br), 2.10 (3H, s), 2.11 (IH, s), 2.50 (IH, m), 2.52 (IH, m), 2.54 (IH, dd, J = 15.0, 3.0 Hz), 2.62 (IH, dd, J = 15.0, 3.8 Hz), 2.81 (IH, dd, J = 5.1, 2.3 Hz), 2.95 (IH, dd, J = 8.3, 2.3 Hz), 3.18 (IH, td, J - 6.5, 4.0 Hz), 3.31 (IH, m), 3.41 (3H, s), 3.53 (IH, br s), 3.76 (IH, m), 5.09 (IH, d, J = 9.1 Hz), 5.15 (IH, d, J = 10.6 Hz), 5.63 (IH, dd, J = 15.2, 9.2 Hz), 5.67 (IH, dd, J = 15.2, 9.1 Hz), 5.70 (IH, dd, J = 15.0, 8.9 Hz), 6.10 (IH, d, J = 10.8 Hz), 6.32 (IH, dd, J = 15.0, 10.7 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 11.0, 11.3, 12.8, 17.4, 17.5, 22.3, 24.7, 25.6, 30.8, 36.2, 39.3, 39.9, 41.7, 43.2, 59.2, 60.3, 60.9, 70.2, 74.4, 75.5, 79.9, 83.6, 84.6, 126.5, 127.3, 132.0, 132.5, 138.2, 141.5, 170.6, 173.0; (+)-HR-ESIMS m/z 589.3413

N1407

¹H NMR (500 MHz, CDCl₃) δθ.88 (3H, d, J = 7.0 Hz), 0.90 (3H, d, J = 6.8 Hz)₅ 0.90 (3H, t, J = 7.1 Hz), 1.15 (3H, d, J = 6.8 Hz), 1.29 (IH, m), 1.31 (3H, br s), 1.40 (IH, m), 1.41 (IH, m), 1.47 (IH, m), 1.51 (IH, m), 1.65 (IH, m), 1.71 (IH, m), 1.75 (3H, s), 1.95 (IH, br s), 2.06 (IH, d, J = 6.0 Hz), 2.50 (IH, m), 2.52 (IH, m), 2.54 (IH, br dd, J = 15.0, 3.1 Hz), 2.61 (IH, dd, J = 15.0, 3.7 Hz), 2.81 (IH, dd, J = 6.0, 2.3 Hz), 2.91 (IH, dd, J = 8.2, 2.3 Hz), 3.18 (IH, td, J = 6.4, 4.0 Hz), 3.31 (IH, ddd, J = 6.0, 6.0, 6.0 Hz), 3.41 (3H, s), 3.54 (IH, br d, J = 10.8 Hz), 3.76 (IH, m), 3.81 (IH, d, J = 8.6 Hz), 5.14 (IH, d, J = 10.7 Hz), 5.70 (IH, dd, J = 15.5, 7.9 Hz), 5.71 (IH, dd, J = 15.5, 8.6 Hz), 5.73 (IH, dd, J = 15.2, 9.8 Hz), 6.08 (IH, d, J = 10.9 Hz), 6.32 (IH, dd, J = 15.2, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.0, 10.3, 11.8, 16.6, 16.6, 23.7, 24.5, 29.8, 35.6, 38.3, 38.9, 40.6, 42.1, 58.1, 59.4, 60.2, 69.3, 73.5, 74.8, 77.2, 82.7, 83.6, 126.3, 130.2, 131.0, 131.5, 137.1, 137.3, 172.2; (+)-HR-ESIMS m/z 547.3220 [M+Na]⁺.

N1512

¹H NMR (500 MHz, CDCl₃) δ 0.89 (3H, d, J = 6.8 Hz), 1.07 (3H, t, J - 7.2 Hz), 1.10 (3H, d, J = 7.3 Hz), 1.15 (3H, d, J = 6.9 Hz), 1.22 (3H, s), 1.33 (IH, m), 1.39 (IH, m), 1.55 (IH, m), 1.71 (IH, m), 1.75 (3H, s), 1.94 (IH, d, J = 6.0 Hz), 2.10 (3H, s), 2.36 (IH, dq, 14.9, 7.3 Hz), 2.50 (IH, m), 2.53 (IH, m), 2.54 (IH, dd, J = 14.5, 2.4 Hz), 2.55 (2H, qd, 7.2, 2.3 Hz), 2.63 (IH, dd, J = 14.5, 3.4 Hz), 2.85 (IH, dd, J = 6.0, 2.2 Hz), 3.03 (IH, dd, J = 8.2, 2.2 Hz), 3.33 (IH, ddd, J = 6.0, 6.0, 6.0 Hz), 3.76 (IH, m), 5.09 (IH, d, J = 9.0 Hz), 5.16 (IH, d, J= 10.7 Hz), 5.62 (IH, dd, J= 15.1, 9.4 Hz), 5.66 (IH, dd, J = 16.0, 7.4 Hz), 5.68 (IH, dd, J - 16.0, 9.0 Hz), 6.09 (IH, d, J = 10.9 Hz), 6.33 (IH, dd, J = 15.1, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 8.4, 12.8, 13.8, 17.4, 17.5, 22.3, 25.6, 30.7, 36.1, 36.5, 39.3, 41.7, 43.2, 49.3, 58.8, 60.4, 70.1, 74.5, 75.2, 79.9, 83.6, 126.6, 127.5, 131.9, 132.7, 137.9, 141.4, 170.7, 173.0, 212.9; (+)-HR-ESIMS m/z 575.3261[M+Na]⁺.

N1513

¹H NMR (500 MHz, CDCl₃) δ 0.90 (3H, d, J = 6.8 Hz), 0.96 (3H, d, J = 7.5 Hz), 0.97 (3H, d, J = 6.8 Hz), 1.02 (3H, d, J = 6.9 Hz), 1.05 (3H, d, J = 6.9 Hz), 1.23 (IH, m), 1.23 (IH, m), 1.37 (IH, m), 1.49 (IH, m), 1.54 (IH, m), 1.75 (3H, s), 1.76 (IH, m), 1.92 (IH, m), 2.03 (3H, s), 2.30 (IH, m), 2.43 (IH, m), 2.48 (IH, m), 2.52 (IH, dd, J = 15.0, 3.0 Hz), 2.60 (IH, dd, J = 15.0, 4.0 Hz), 3.39 (IH, m), 3.70 (IH, m), 4.00 (IH, dd, J = 6.5, 6.2 Hz), 4.96 (IH, dd, J = 10.1, 9.9 Hz), 5.15 (IH, d, J = 10.6 Hz), 5.35 (IH, dd, J = 15.0, 9.9 Hz), 5.51 (IH, dd, J = 15.5, 6.5 Hz), 5.56 (IH, dd, J = 15.0, 10.0 Hz), 5.64 (IH, dd, J == 15.5, 8.0 Hz), 5.69 (IH, dd, J = 15.2, 8.1 Hz), 6.10 (IH, d, 10.8 Hz), 6.27 (IH, dd, J = 15.2, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.4, 11.8, 14.7, 15.6, 15.7, 16.6, 21.4, 26.8, 27.9, 28.0, 34.9, 38.6, 40.6, 41.9, 43.3, 69.6, 76.2, 76.3, 79.0, 82.7, 126.1, 129.0, 129.0, 131.1, 131.1, 135.2, 138.0, 138.9, 170.4, 172.0; (+)-HR-ESIMS m/z 543.3296 [M+Na]⁺.

N1514

¹H NMR (500 MHz, CDCl₃) δ 0.89 (3H, d, J = 6.8 Hz), 0.90 (3H, d, J = 7.6 Hz), 1.15 (3H, d, J = 7.3 Hz), 1.16 (3H, d, J = 6.5 Hz), 1.22 (3H, s), 1.28 (IH, m), 1.35 (IH, qd, J = 7.6, 4.3 Hz), 1.38 (IH, m), 1.55 (IH, m), 1.71 (IH, m), 1.74 (3H, s), 2.10 (3H, s), 2.49 (IH, m), 2.52 (IH, m), 2.54 (IH, br dd, J = 15.0, 3.0 Hz), 2.63 (IH, dd, J = 15.0, 3.6 Hz), 2.81 (IH, dd, J = 5.6, 2.3 Hz), 2.91 (IH, dd, J = 8.2, 2.3 Hz), 3.32 (IH, dd, J = 6.0, 5.6 Hz), 3.36 (3H, s), 3.42 (IH, qd, J = 6.5, 4.3 Hz), 3.75 (IH, m), 5.09 (IH, d, J = 9.2 Hz), 5.16 (IH, d, J = 10.7 Hz), 5.62 (IH, dd, J = 15.3, 9.4 Hz), 5.65 (IH, dd, J = 15.3, 9.2 Hz), 5.69 (IH, dd, J = 15.1, 8.4 Hz), 6.09 (IH, d, J = 10.9 Hz), 6.32 (IH, dd, J = 15.1, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 11.7, 12.8, 17.1, 17.4, 17.5, 22.3, 25.6, 30.8, 36.2, 39.3, 41.7, 42.6, 43.3, 57.5, 60.0, 60.4, 70.2, 74.4, 75.5, 79.0, 79.9, 83.6, 126.5, 127.3, 132.0, 132.5, 138.2, 141.5, 170.6, 173.0; (+)-HR-ESIMS m/z 575.3282 [MH-Na]⁺. N1515

¹H NMR (500 MHz, CDCl₃) δ 0.82 (3H, d, J = 6.8 Hz), 0.87 (3H, d, J = 7.3 Hz), 0.90 (3H, t, J = 7.2 Hz), 1.15 (3H, d, J = 6.8 Hz), 1.32 (3H, s), 1.40 (IH, qd, J = 7.3, 4.1 Hz), 1.47 (IH, m), 1.64 (IH, m), 1.74 (3H, s), 2.11 (3H, s), 2.38 (IH, m), 2.49 (IH, m), 2.49 (IH, dd, J = 14.0, 2.4 Hz), 2.71 (IH, dd, J = 14.0, 5.4 Hz), 2.81 (IH, dd, J = 5.4, 2.3 Hz), 2.90 (IH, dd, J = 8.2, 2.3 Hz), 3.17 (IH, td, J = 6.4, 4.1 Hz), 3.30 (IH, dd, J = 6.7, 5.4 Hz), 3.40 (3H, s), 4.53 (IH, m), 5.04 (IH, d, J = 9.2 Hz), 5.09 (IH, d, J - 10.4 Hz), 5.38 (IH, dd, J = 15.2, 9.5 Hz), 5.47 (IH, dd, J = 15.2, 9.2 Hz), 5.58 (IH, dd, J = 15.5, 1.8 Hz), 5.68 (IH, dd, J = 15.1, 8.6 Hz), 5.81 (IH, dd, J = 15.5, 2.7 Hz), 6.02 (IH, d, J = 10.9 Hz), 6.31 (IH, dd, J =

15.1, 10.9 Hz),; ¹³C NMR (125 MHz, CDCl₃) δ 11.4, 11.4, 12.8, 17.6, 18.1, 22.4, 24.9, 27.2,

40.2, 41.2, 41.5, 43.3, 59.2 60.3, 61.0, 68.5, 75.7, 75.4, 80.8, 84.3, 84.9, 126.6, 127.5, 131.8, 132.2, 132.4, 132.6, 138.5, 141.0, 170.5, 172.5; (÷)-HR-ESIMS m/z 588.3828 [M+Na]⁺.

N1516

¹H NMR (500 MHz, CDCl₃) δ 0.85 (3H, d, J = 6.8 Hz), 0.88 (3H, d, J = 7.3 Hz), 0.90 (3H, t, J = 7.7 Hz), 1.15 (3H, d, J = 6.8 Hz), 1.36 (3H, s), 1.41 (IH, dq, J = 7.3, 3.8), 1.47 (IH, m), 1.65 (IH, m), 1.73 (3H, s), 2.16 (3H, s), 2.37 (IH, m), 2.50 (IH, m), 2.70 (IH, dd, J = 16.2, 3.8 Hz), 2.78 (IH₃ dd, J = 16.2, 3.3 Hz), 2.81 (IH, dd, J = 5.6, 2.2 Hz), 2.85 (IH, dd, J = 19.6, 10.7 Hz), 2.91 (IH, dd, J = 8.2, 2.2 Hz), 3.06 (IH, dd, J = 19.6, 3.2 Hz), 3.18 (IH, td, J = 6.4, 3.8 Hz), 3.33 (IH, ddd, 6.0, 6.0, 6.0 Hz), 3.41 (3H, s), 4.28 (IH, m), 5.13 (IH, d, J = 10.5 Hz), 5.42 (IH, dd, J = 15.0, 9.7 Hz), 5.50 (IH, d, J = 9.7 Hz), 5.70 (IH, dd, J = 15.0, 8.7 Hz), 5.74 (IH, dd, J = 15.0, 10.1 Hz), 6.06 (IH, d, J = 10.8 Hz), 6.31 (IH, dd, J = 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 1 1.0, 11.3, 12.8, 17.1, 17.4, 22.2, 24.6, 24.7, 38.9, 39.9, 41.9, 43.2, 43.2, 59.1, 60.2, 60.8, 65.5, 75.7, 77.8, 80.9, 83.6, 84.6, 124.7, 127.2, 132.0, 132.1, 138.5, 144.3, 171.4, 173.3, 209.8; (+)-HR-ESIMS m/z 603.3150 [MH-Na]⁺.

N1519

¹H NMR (500 MHz, CDCl₃) δθ.82 (3H, d, J = 6.8 Hz), 1.07 (3H, t, J = 7.3 Hz), 1.10 (3H, d, J = 7.1 Hz), 1.15 (3H, d, J = 6.8 Hz), 1.32 (3H, s), 1.75 (3H, s), 2.12 (3H, s), 2.35 (IH, m), 2.39 (IH, m), 2.50 (IH, m), 2.50 (IH, m), 2.55 (2H, m), 2.72 (IH, dd, J = 14.1, 5.4 Hz), 2.85 (IH, dd, J = 5.0, 2.2 Hz), 3.03 (IH, dd, J = 8.2, 2.2 Hz), 3.33 (IH, dd, J = 7.0, 5.0 Hz), 4.53 (IH, m), 5.04 (IH, d, J = 9.3 Hz), 5.10 (IH, d, J = 10.4 Hz), 5.39 (IH, dd, J = 15.2, 9.3 Hz), 5.48 (IH, dd, J = 15.2, 9.3 Hz), 5.59 (IH, dd, J = 15.5, 1.8 Hz), 5.66 (IH, dd, J = 15.1, 8.7 Hz), 5.81 (IH, dd, J = 15.5, 2.7 Hz), 6.02 (IH, d, J = 10.8 Hz), 6.31 (IH, dd, J = 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 8.5, 12.8, 13.9, 17.5, 18.1, 22.3, 27.1, 36.6, 41.2, 41.5, 43.3, 49.5, 58.7, 60.4, 68.5, 75.2, 75.4, 80.8, 84.0, 126.7, 127.6, 131.6, 132.3, 132.4, 132.9, 138.0, 140.9, 170.7, 172.5, 212.8; (+)-HR-ESIMS m/z, 571.2907 [M+Naf.

N1523

¹H NMR (500 MHz, CDCl₃) δ 0.89 (3H, d, J = 6.8 Hz), 0.94 (3H, d, J = 7.3 Hz), 0.98 (3H₅ t, J = 7.4 Hz), 1.16 (3H, d, J = 6.8 Hz), 1.22 (3H, s), 1.33 (IH, m), 1.40 (IH, m), 1.45 (IH, qd, J - 7.3, 3.7 Hz), 1.55 (IH, m), 1.57 (IH, m), 1.72 (IH, m), 1.75 (3H, s), 2.10 (3H, s), 2.51 (IH, m), 2.54 (IH, m), 2.54 (IH, dd, J = 15.0, 3.0 Hz), 2.63 (IH, dd, J = 15.0, 3.6 Hz), 2.88 (IH, dd, J = 5.1, 2.3 Hz), 2.94 (IH, dd, J = 7.7, 2.3 Hz), 3.32 (IH, dd, J = 5.1, 6.8 Hz), 3.62 (IH, m), 3.76 (IH, m), 5.09 (IH, d, J = 9.2 Hz), 5.16 (IH, d, J = 10.7 Hz), 5.62 (IH, dd, J = 15.2, 9.4 Hz), 5.68 (IH, dd, 15.2, 9.2 Hz), 5.69 (IH, dd, J = 15.0, 8.7 Hz), 6.09 (IH, d, J = 10.8 Hz), 6.32 (IH, dd, J = 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.4, 10.5, 11.8, 16.4, 16.6, 21.3, 24.6, 27.3, 29.8, 35.2, 38.4, 40.2, 40.7, 42.2, 59.6, 59.8, 69.2, 73.4, 74.4, 74.6, 78.9, 82.7, 125.6, 126.4, 131.0, 131.6, 137.2, 140.5, 169.7, 172.1; (+)-HR- ESIMS m/z 575.3261 [M+Na]⁺.

N1551

¹H NMR (500 MHz, CDCl₃) δ 0.87 (3H, d, J = 6.8 Hz), 0.92 (3H, d, J = 7.1 Hz), 0.96 (3H, t, J = 6.9 Hz), 0.97 (3H, d, J = 6.5 Hz), 1.14 (3H, d, J = 6.8 Hz), 1.23 (IH, m), 1.23 (IH, m), 1.44 (IH, m), 1.47 (IH, m), 1.51 (2H, m), 1.74 (3H, s), 1.75 (IH, m), 1.90 (IH, m), 2.02 (3H, s), 2.48 (IH, m), 2.49 (IH, m), 2.51 (IH, dd, J = 14.8, 3.0 Hz), 2.59 (IH, dd, J = 14.8, 3.7 Hz), 2.87 (IH, dd, J = 5.3, 2.3 Hz), 2.93 (IH, dd, J = 7.7, 2.3 Hz), 3.28 (IH, dd, J = 7.0, 5.3 Hz), 3.61 (IH, td, J = 6.5, 3.5 Hz), 3.70 (IH, m), 4.96 (IH, dd, J = 10.1, 9.8 Hz), 5.13 (IH, d, J = 10.6 Hz), 5.35 (IH, dd, J = 15.0, 9.8 Hz), 5.54 (IH, dd, J = 15.0, 10.0 Hz), 5.68 (IH, dd, J = 15.0, 8.7 Hz), 6.09 (IH, br d, J = 10.8 Hz), 6.31 (IH, dd, J = 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.4, 10.5, 11.8, 15.6, 16.5, 16.6, 21.4, 27.2, 27.9, 28.0, 34.9, 38.6, 40.2, 40.2, 42.2, 59.7, 59.8, 69.6, 74.4, 74.6, 78.9, 82.6, 126.4, 129.1, 130.9, 131.7, 137.1, 138.8, 170.4, 172.0; (+)-HR-ESIMS m/z 559.3325 [M+Naf.

N1611

¹H NMR (500 MHz, CDCl₃) δ 0.88 (3H, d, J = 7.0 Hz), 0.89 (3H, d, J = 7.0 Hz), 0.92 (3H, t, J = 7.2 Hz), 1.15 (3H, d, J = 8.0 Hz), 1.16 (3H, s), 1.41 (IH, m), 1.42 (IH, m), 1.46 (IH, m), 1.68 (IH, m), 1.75 (IH, m), 1.76 (3H, s), 2.10 (3H, s), 2.50 (IH, m), 2.51 (IH, m), 2.63 (IH, dd, J = 15.6, 3.0 Hz), 2.69 (IH, dd, J = 15.6, 4.0 Hz), 2.81 (IH, dd, J = 5.2, 2.3 Hz), 2.90 (IH, dd, J = 8.2, 2.3 Hz), 3.18 (IH, td, J = 6.4, 3.9 Hz), 3.32 (IH, m), 3.40 (3H, s), 3.57 (IH, dd, J = 11.3, 11.3 Hz), 4.17 (IH, m), 5.08 (IH, d, J = 9.3 Hz), 5.18 (IH, d, J = 10.6 Hz), 5.59 (IH, m), 5.59 (IH, m), 5.70 (IH, dd, J = 15.1, 8.5 Hz), 6.09 (IH, d, J = 10.9 Hz), 6.32 (IH, dd, J = 15.1, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.0, 10.3, 11.9, 16.4, 16.5, 18.4, 21.3, 23.7, 35.2, 37.6, 38.9, 40.9, 42.3, 58.2, 59.3, 59.9, 66.3, 71.5, 74.6, 76.4, 78.1, 82.4, 83.6, 124.8, 126.3, 131.1, 131.3, 137.4, 141.3, 169.5, 172.3; (+)-HR-ESIMS m/z

Example 5: Synthesis of Macrolide Compounds

The compounds of this invention, and related derivatives, can be synthesized by methods known to one skilled in the art. Schemes 1-13, shown below, depict the synthesis of several macrolide compounds of the invention with various degrees of substitution. Characterization and detailed methods for synthesizing these compounds are described below.

Scheme 1 Procedures for the Syntheses of Halohydrins

Synthesis of Bromohydrins:

To a solution of N831 (A) (202 mg, 0.36 mmol) in anhydrous 1,2-dirnethoxyethane ether (DME)-ether 1:1 (5 niL) at 0 ⁰C was added 48% HBr (100 μL, 0.6 mmol). The reaction mixture was stirred at 0 ⁰C for 1.5 hours. Additional 48% HBr (100 μL, 0.6 mmol) was introduced to the mixture twice over a period of 20 min. The excess HBr was neutralized by adding K₂CO₃ and saturated aqueous NaHCO₃ (0.3 mL). The stirring was continued at 0 ⁰C for 5 min. The mixture was then warmed to room temperature, dried with anhydrous MgSO₄ and then filtered. The residue was rinsed further with ethyl acetate (3 x 15 mL). The filtrates were combined and concentrated under reduced pressure. The crude product was purified by preparative TLC (chloroform-methanol 25:1, developed twice) to give GM120 as a colourless oil (23.6 mg, 10%) and GM121 as a colourless oil (100.6 mg, 43%).

GM120

¹H NMR (500 MHz, CDCl₃) £0.90 (3 H, t, J= 7.7 Hz), 0.91 (3H, t, J= 7.2 Hz), 1.08 (3H, d, J = 7.3 Hz), 1.10 (3H, d, J - 6.8 Hz), 1.23 (3H, s), 1.33 (IH, m), 1.41 (IH, m), 1.42 (IH, m), 1.56 (2H, m), 1.69 (IH, m), 1.76 (3H, s), 1.84 (IH, m), 2.11 (3H, s), 2.48 (IH, m), 2.49 (IH, m), 2.54 (IH, m), 2.63 (IH, dd, J= 15.0, 3.3 Hz), 2.99 (IH, m), 3.38 (3H, s), 3.55 (IH, m), 3.76 (IH, m), 3.88 (IH, dd, J = 9.8, 7.9 Hz), 3.95 (IH, m), 4.01 (IH, dd, J = 7.9, 3.4 Hz), 5.10 (IH, d, J = 9.0 Hz), 5.19 (IH, d, J = 10.7 Hz), 5.63 (IH, dd, J = 15.2, 9.3 Hz), 5.68 (IH, dd, J= 15.2, 9.0 Hz), 5.92 (IH, dd, J= 15.0, 7.8 Hz), 6.14 (IH, d, J = 10.6 Hz), 6.32 (IH, dd, J = 15.0, 10.6 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.7, 11.7, 12.8, 13.6, 17.5, 22.3, 22.9, 25.6, 30.8, 35.8, 36.2, 39.3, 41.8, 41.9, 56.3, 57.0, 70.2, 74.4, 79.3, 79.9, 82.2, 83.7, 83.8, 125.9, 126.4, 131.5, 132.5, 141.1, 141.6, 170.6, 173.0; (+)-HR-ESIMS m/z 669^2582 [M+Na]⁺, 671.2625 [M+Na]⁺

GM121

¹H NMR (500 MHz, CDCl₃) 50.88 (3H, d, J = 6.7 Hz), 0.91 (3H, t, J= 7.4 Hz), 1.11 (3H, d, J= 7.1 Hz), 1.16 (3H, d, J= 6.7 Hz), 1.23 (3H, s), 1.32 (IH, m), 1.39 (IH, m), 1.55 (IH, m), 1.59 (2H, m), 1.67 (IH, m), 1.74 (3H, s), 2.11 (3H, s), 2.34 (IH, m), 2.52 (H-16, 2H, m), 2.55 (IH, m), 2.64 (IH, dd, J= 15.0, 3.6 Hz), 3.31 (IH, m), 3.34 (3H, s), 3.77 (IH, m), 3.81 (2H, m), 4.29 (IH, dd, J= 9.8, 2.3 Hz), 5.10 (IH, d, J= 9.1 Hz), 5.17 (IH, d, J= 10.7 Hz), 5.62 (IH, dd, J = 15.3, 9.0 Hz), 5.65 (IH, m), 5.68 (IH, dd, J= 15.3, 9.1 Hz), 6.11 (IH, d, J= 10.8 Hz), 6.32 (IH, dd, J= 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.3, 12.7, 14.1, 17.4, 18.4, 22.3, 23.5, 25.6, 30.8, 36.2, 39.3, 40.6, 41.7, 42.6, 57.9, 60.9, 70.2, 73.5, 74.4, 75.5, 79.9, 83.6, 84.2, 126.6, 127.0, 132.0, 132.1, 139.2, 141.5, 170.6, 173.1; (+)-HR-ESIMS m/z 669.2579 [M+Naf, 671.2535 [M+Na]⁺

Synthesis of Cyclic Carbonate Bromohydrin (GMl 23)

19%

To the dried GM121 (7.8 mg, 0.01 mmol) was added a solution of triphosgene (11 mg, 0.04 mmol) in anhydrous CH₂Cl₂ (1 mL) and triethylamine (15 μL, 0.11 mmol). The reaction mixture was stirred at room temperature for 2 hours. Additional triphosgene (11 mg, 0.04 mmol) and triethylamine (15 μL, 0.11 mmol) were added. The mixture was stirred at room temperature for another hour before being quenched with saturated aqueous NaHCO₃ (2 mL). The mixture was extracted with CH₂Cl₂ (2 x 5 mL). The organic extracts were combined, dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by preparative TLC (chloroform-methanol 50:1) to give GM123 as a colorless oil (1.5 mg, 19%). GM123

¹H NMR (500 MHz, CDCl₃) 50.91 (3H, d, J= 6.8 Hz), 0.94 (3H, t, J= 7.4 Hz), 1.09 (3H, d, J= 6.6 Hz), 1.19 (3H, d, J= 6.9 Hz), 1.23 (3H, s), 1.32 (IH, m), 1.40 (IH, m), 1.51 (IH, m), 1.56 (IH, m), 1.67 (IH, m), 1.69 (IH, m), 1.77 (3H, s), 2.11 (3H, s), 2.21 (IH, m), 2.53 (IH, m), 2.56 (IH, m), 2.64 (H-16, 2H, m), 3.06 (IH, ddd, J= 6.7, 6.7, 3.9 Hz), 3.36 (3H, s), 3.76 (IH, m), 4.08 (IH, dd, J= 9.8, 2.6 Hz), 4.52 (IH, dd, J= 9.8, 3.2 Hz), 4.56 (IH, dd, J= 5.1, 3.2 Hz), 5.10 (IH, d, J= 9.3 Hz), 5.18 (IH, d, J= 10.6 Hz), 5.62 (IH, dd, J= 15.2, 9.0 Hz), 5.62 (IH, dd, J = 15.1, 9.0 Hz), 5.69 (IH, dd, J = 15.2, 9.3 Hz), 6.10 (IH, d, J = 10.8 Hz), 6.38 (IH, dd, J= 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) 510.1, 12.5, 12.9, 15.6, 17.4, 22.3, 23.5, 25.6, 30.8, 36.2, 37.0, 39.3, 41.7, 42.0, 57.8, 58.6, 70.2, 74.4, 78.5, 79.9, 83.4, 84.5, 85.3, 126.7, 129.2, 131.1, 134.1, 134.5, 141.4, 154.4, 170.6, 173.0; (+)- HR-ESIMS m/z 695.2380 [M+Na]⁺, 697.2486 [M+Naf.

Synthesis of Chlorohydrin (GM122)

G Wl 122

39%

To a suspension of WCl₆ (198 mg, 0.5 mmol) in anhydrous THF (5 mL) at -78 ⁰C was added H-BuLi (1.6 M in hexane; 0.625 mL, 1.0 mmol). The suspension was wanned to room temperature slowly with continuous stirring to give a deep blue solution, which was added to N831 (A) (11 mg, 0.02 mmol). The reaction mixture was stirred at room temperature for 24 hours before being quenched with saturated aqueous NaHCO₃ (2 niL). The mixture was partitioned between ethyl acetate (5 mL) and water (3 mL), and extracted with ethyl acetate (2 x 5 mL). The organic extracts were combined, dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by preparative TLC (chloroform-methanol 25:1, developed twice) to give the GM122 as a colorless oil (4.5 mg, 39%).

GM122

¹H NMR (500 MHz, CDCl₃) £0.88 (3H, d, J = 6.7 Hz), 0.92 (3H, t, J= 7.4 Hz), 1.09 (3H, d, J= 7.1 Hz), 1.16 (3H, d, J= 6.7 Hz), 1.23 (3H, s), 1.32 (IH, m), 1.39 (IH, m), 1.55 (IH, m), 1.62 (2H, m), 1.70 (IH, m), 1.74 (3H, s), 2.11 (3H, s), 2.35 (IH, m), 2.52 (2H, m), 2.55 (IH, m), 2.64 (IH, dd, J= 15.0, 3.4 Hz), 3.28 (IH, m), 3.34 (3H, s), 3.69 (IH, m), 3.76 (2H, m), 4.15 (IH, dd, J= 9.6, 2.2 Hz), 5.10 (IH, d, J= 9.1 Hz), 5.17 (IH, d, J= 10.7 Hz), 5.62 (IH, dd, J= 15.0, 9.0 Hz), 5.62 (IH, dd, J= 15.0, 9.0 Hz), 5.70 (IH, dd, J= 15.0, 9.1 Hz), 6.11 (IH, d, J= 10.9 Hz), 6.32 (IH, dd, J= 15.0, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.6, 12.7, 12.9, 17.4, 18.3, 22.3, 23.6, 25.6, 30.8, 36.2, 39.3, 40.1, 41.7, 42.4, 57.8, 65.1, 70.2, 73.5, 74.4, 74.8, 79.9, 83.6, 83.9, 126.6, 127.0, 132.06, 132.11, 139.2, 141.5, 170.6, 173.1; (+)-HR-ESIMS m/z 625.2614 [M+Na]⁺ 627.2877 [M+Na]⁺

Synthesis of Iodohydrin (GM129)

GM129

22% A mixture of N831 (A) (10.7 mg, 0.02 mmol), LiI (20.1 mg, 0.15 mmol) in dimethoxyethane ether (7 mL) was heated to 68 ⁰C and stirred under argon for 5 days. The solvent was evaporated and the residue partitioned between ethyl acetate and saturated aqueous Na₂S₂O₃ solution. The ethyl acetate extract was washed with water, dried with anhydrous MgSO₄ and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (gradient elution with 50% ethyl acetate-hexane to 100% ethyl acetate, then 5% methanol-chloroform) to give GM129 (2.9 mg, 22%).

GM129

¹H NMR (500 MHz, CDCl₃) £0.87 (3H, t, J = 6.8 Hz), 0.88 (3H, d, J= 7.4 Hz), 0.89 (3H, d, J= 7.1 Hz), 1.14 (3H, d, J= 6.7 Hz), 1.22 (3H, s), 1.30 (IH, m), 1.99 (IH, m), 1.38 (IH, m), 1.56 (IH, m), 1.60 (2H, m), 1.70 (IH, m), 1.73 (3H, s), 2.09 (3H₅ s), 2.49 (IH, m), 2.51 (IH, m), 2.55 (IH, dd, J= 15.1, 3.5 Hz), 2.61 (IH, dd, J= 15.1, 2.9 Hz), 3.24 (IH, m), 3.34 (3H, s), 3.75 (IH, m), 3.88 (IH, m), 3.91 (IH, m), 4.43 (IH, d, J= 10.0 Hz ), 5.09 (IH, d, J = 9.0 Hz), 5.17 (IH, d, J= 10.6 Hz), 5.62 (IH, dd, J= 15.2, 9.1 Hz), 5.63 (IH, dd, J= 15.0, 8.4 Hz), 5.65 (IH, dd, J = 15.2, 9.1 Hz), 6.10, (IH, d, 10.8), 6.30 (IH, dd, Jdd, 15.0, 10.8); (+)-HR-ESIMS m/z 717.2423 [M+Naf

Scheme 2 Procedure for the Syntheses of N831 (A) Rn Ketone and Epimer

diTES N831

GM137

43%

63%

To a solution of N831 (A) (960 mg, 1.7 mmol), DMAP (104 mg, 0.85 mmol) and triethylamine (0.236 mL, 1.7 mmol) in anhydrous THF (32 mL) was added chlorotriethylsilane (TES-Cl) (0.285 mL, 1.7 mmol). The reaction mixture was stirred at room temperature for 1 hour. Additional chlorotriethylsilane (0.029 mL, 0.17 mmol) and triethylamine (0.024 mL, 0.17 mmol) were introduced. The mixture was stirred at room temperature for another 30 min and then quenched with water (8 mL). The hulk of THF was removed under reduced pressure. The residue was extracted with ethyl acetate (2 x 15 mL). The organic extracts were combined, dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (gradient elution with ethyl acetate-hexane 1:3 to ethyl acetate only) to give compound GM137 as a colourless oil (499 mg, 43%).

To a solution of GM137 (11.5 mg, 0.02 mmol) in dry CH₂Cl₂ (1 mL) was added Dess-Martin periodinane (24 mg, 0.06 mmol) at room temperature under an argon atmosphere. The reaction mixture was stirred at room temperature until all starting material had reacted as indicated by TLC (ethyl acetate-hexane 1 : 1). It was worked-up by washing successively with saturated aqeous NaHCO₃ and brine, dried (MgSO₄), filtered and concentrated. The crude product was purified by flash column chromatography (hexane- ethyl acetate 1:1) to give ketone 1 (7 mg, 61%). To ketone 1 (5 mg, 7.37 μmol) in dry MeOH-CH₂Cl₂ (1:1, 2 mL) was added pyridiniump-toluenesulfonate (3 mg, 0.0119 mmol). The solution was stirred at room temperature until all starting material had reacted as indicated by TLC (ethyl acetate-hexane 1:1). The solvents were then removed under reduced pressure and the crude product purified by flash column chromatography (ethyl acetate-hexane 3:1 then 4:1) to give GM144 (3 mg, 72%).

To a solution of dry THF (1.3 mL) was added (^-2-methyl-CBS-oxazaborolidine (IM in toluene, 20 μL, 0.02 mmol) and catechol borane (IM in THF, 65 μL, 0.07 mmol). It was stirred for 10 min at -78 ⁰C prior to the dropwise addition of ketone 1 (6.6 mg, 9.73 μmol) dissolved in dry THF (1 mL). The reaction mixture was left at -78 ⁰C for 10 min, then 0 ⁰C (2 hours) and finally at room temperature overnight. It was worked up by first adding methanol, stirred for 10 min and then concentrated under reduced pressure. The crude product was purified by flash column chromatography (ethyl acetate-hexane 1 :2) to give alcohol 2 (2.3 mg, 35%). To a cooled solution of 2 (2.3 mg, 3.38 μmol) in dry methanol (1 mL) was added pyridinium/7-toluenesulfonate (5.2 mg, 20.7 μmol). The reaction mixture was stirred at 0 ⁰C until all starting material had reacted as indicated by TLC (ethyl acetate-hexane 1:1). The solvent was then removed under reduced pressure and the crude product purified by flash column chromatography (ethyl acetate-hexane 2:1, 4:1, then 9:1) to give GM158 (1.2 mg, 63%). 3M137

¹H NMR (500 MHz, CDCl₃) £0.62 (6H, m), 0.88 (3H, m), 0.89 (3H, m), 0.91 (3H, m), 0.98 (9H, m), 1.14 (3H, d, J- 6.8 Hz), 1.21 (3H, s), 1.42 (IH, m), 1.46 (IH, m), 1.47 (IH, m), 1.48 (2H, m), 1.65 (IH, m), 1.69 (IH, m), 1.71 (3H₉ s), 2.10 (3H, s), 2.41 (IH, dd, J= 13.7, 4.8 Hz), 2.46 (IH, dd, J= 13.7, 3.5 Hz), 2.48 (IH, m), 2.52 (IH, m), 2.81 (IH, dd, J= 5.2, 2.1 Hz), 2.90 (IH, dd, J= 8.3, 2.1 Hz), 3.18 (IH, ddd, J= 6.4, 6.4, 4.1 Hz), 3.30 (IH, dd, J = 6.7, 5.2 Hz), 3.41 (3H, s), 3.85 (IH, m), 4.96 (IH, d, J= 10.8 Hz), 5.08 (IH, d, J= 8.8 Hz), 5.65 (IH, m), 5.65 (IH, m), 5.66 (IH, m), 6.10 (IH, d, J= 10.7 Hz), 6.31 (IH, dd, J = 15.0, 10.7 Hz); (+)-HR-ESIMS m/z 703.4260 [MH-Na]⁺

GM144

¹H-NMR (500MHz, CDCl₃) 50.90 (3H, d, J= 7.0 Hz), 0.90 (3H, d, J= 6.9 Hz), 0.92 (3H, t, J = 7.2 Hz), 1.22 (3H, s), 1.24 (3H, d, J= 5.1 Hz), 1.28 (IH, m), 1.39 (IH, m), 1.50 (IH, m), 1.56 (IH, m), 1.58 (IH, m), 1.65 (IH, m), 1.73 (IH, m), 1.77 (3H, s), 2.10 (3H, s), 2.53 (IH, m), 2.56 (IH, dd, J= 15.1, 2.8 Hz), 2.61 (IH, dd, J= 15.1, 3.6 Hz), 3.02 (IH, dd, J = 7.5, 1.8 Hz), 3.20 (IH, m), 3.36 (IH, d, J= 1.8 Hz), 3.41 (3H, s), 3.43 (IH, m), 3.49 (IH, d, J= 10.9 Hz), 3.76 (IH, m), 5.09 (IH, d, J= 9.2 Hz), 5.16 (IH, d, J= 11.6 Hz), 5.63 (IH, dd, J= 15.1, 9.5 Hz), 5.67 (IH, m), 5.71 (H-15, IH, m), 6.10 (IH, d, J= 10.8 Hz) 6.39 (IH, dd, J= 10.8, 15.2 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.0, 12.9, 16.5, 17.4, 22.2, 24.5, 25.6, 30.8, 36.2, 39.3, 39.9, 41.7, 47.0, 58.9, 59.0, 62.0, 70.1, 74.4, 79.8, 83.4, 84.4, 126.6, 129.1, 131.3, 133.8, 133.9, 141.3, 170.6, 172.9, 207.0; (+)-HR-ESIMS m/z 587.3183 [MrHNa]⁺

GM158

¹H-NMR (500MHz, CDCl₃) £0.89 (3H, d, J = 7.1 Hz), 0.92 (3H, t, J= 7.6 Hz), 0.92 (3H, d, J = 7.0 Hz), 1.16 (3H, d, J= 6.9 Hz), 1.22 (3H, s), 1.28 (IH, m), 1.38 (IH, m), 1.46 (IH, m), 1.50 (IH, m), 1.55 (IH, m), 1.67 (IH, m), 1.71 (IH, m), 1.75 (3H, s), 2.10 (3H, s), 2.51 (2H, m), 2.54 (IH, m), 2.62 (IH, dd, J= 15.6, 3.6 Hz), 2.81 (IH, m), 3.00 (IH, dd, J= 8.1, 2.1 Hz), 3.19 (IH, m), 3.41 (3H, s), 3.53 (IH, d, J= 11.0 Hz), 3.64 (IH, m), 3.79 (IH, m), 5.09 (IH, d, J = 9.0 Hz), 5.16 (IH, d, J = 10.6 Hz), 5.61 (IH, dd, J= 15.0, 9.3 Hz), 5.65 (IH, dd, 15.0, 9.0 Hz), 5.80 (IH, dd, J= 15.1, 8.4 Hz), 6.11 (IH, d, J= 10.6 Hz), 6.31 (IH, dd, J = 15.1, 10.6 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.4, 12.7, 17.5, 17.7, 22.2, 24.7, 25.6, 30.8, 36.2, 39.3, 39.7, 41.7, 42.2, 58.3, 59.0, 60.2, 70.1, 73.4, 74.4, 79.9, 83.6, 84.7, 126.5, 127.3, 132.0, 132.2, 138.2, 141.5, 170.6, 173.0; (+)-HR-ESIMS m/z 589.3377 [M+Na]⁺

Scheme 3 Procedure for the Synthesis of N831 (A) Ri _fi AIlylic Alcohol

GM171 (23%)

To a solution of N831 (A) (13.2 mg, 0.02 mmol) in anhydrous THF (5 mL) was added SeO₂ (50 mg, 0.45 mmol) at room temperature. The reaction mixture was then heated at reflux until all starting material had reacted as indicated by TLC (chloroform-methanol 19: 1). The solvent was removed under reduced pressure and the crude product purified by preparative TLC (chloroform-methanol 19:1 then 9:1) to give GM171 (3.4 mg, 23%).

GM171

¹H-NMR (500MHz, CDCl₃) £0.90 (3H, t, J= 7.5 Hz), 0.90 (3H, d, J= 7.0 Hz), 0.90 (3H, d, J= 6.5 Hz), 1.22 (3H, s), 1.28 (IH, m), 1.38 (IH, m), 1.38 (3H₅ s), 1.46 (IH, m), 1.47 (IH, m), 1.55 (IH, m), 1.66 (IH, m), 1.71 (IH, m), 1.79 (3H, s), 2.10 (3H, s), 2.26 (IH, d, J= 6.9 Hz), 2.53 (IH, m), 2.56 (IH, m), 2.62 (IH, dd, J = 14.8, 3.7 Hz), 2.95 (IH, m), 2.96 (IH, m), 3.18 (IH, ddd, J= 6.5, 4.0, 3.9 Hz), 3.40 (IH^ m), 3.41 (3H, s), 3.52 (IH, d, J= 10.8 Hz), 3.76 (IH, m), 5.09 (IH, d, J = 9.3 Hz), 5.18 (IH, d, J= 10.7 Hz), 5.62 (IH, dd, J = 15.2, 9.4 Hz), 5.67 (IH, dd, J= 15.2, 9.3 Hz), 5.85 (H-15, IH, d, J= 15.1 Hz), 6.15 (IH, d, J= 11.0 Hz), 6.61 (IH, dd, J= 15.1, 11.0 Hz); (Hr)-HR-ESIMS m/z 605.3322 [M+Na]⁺

Scheme 4 Procedure for Methylation at R31

N1512 GM166

37%

To a solution of N1512 (4.5 mg, 8.2 μmol) in dry THF (2 mL) at 0 ⁰C was added MeMgCl (3 M in THF; 5.4 μL, 16.4 μmol) portion-wise at 0 ⁰C over a period of 75 min. The reaction mixture was quenched with saturated aqueous NH₄Cl (0.2 mL), dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by preparative TLC (chloroform-methanol 20:1) to give the GM166 as a colourless oil (1.7 mg, 37%). GM166 (a mixture of R₂i-epimers in a ratio of 45:55)

¹H NMR (500 MHz, CDCl₃) (minor epimer) £0.90-0.92 (6H, m), 0.95 (3H, t, J= 7.5 Hz), 1.17 (3H, d, J= 6.8 Hz), 1.21 (3H, s), 1.23 (3H, s), 1.32 (IH, m), 1.39 (IH, m), 1.40 (IH, m), 1.54 (3H, m), 1.72 (IH, m), 1.76 (3H, s), 1.97 (IH, br s), 2.11 (3H, s), 2.52 (2H, m), 2.54 (IH, m), 2.64 (IH, dd, J= 14.9, 3.8 Hz), 2.83 (IH, dd, J= 4.9, 2.3 Hz), 2.99 (IH, dd, J = 8.6, 2.3 Hz), 3.34 (IH, m), 3.53 (IH, d, J= 10.9 Hz), 3.77 (IH, m), 5.10 (IH, d, J= 9.2 Hz), 5.17 (IH, d, J= 10.7 Hz), 5.63 (IH, dd, J= 15.2, 9.3 Hz), 5.69 (2H, m), 6.10 (IH, d, J = 10.8 Hz), 6.33 (IH, dd, J= 15.1, 10.8 Hz)

¹H NMR (500 MHz, CDCl₃) (major epimer) £0.90-0.92 (6H, m), 0.95 (3H, t, J = 7.5 Hz), 1.16 (3H, d, J= 6.8 Hz), 1.20 (3H, s), 1.23 (3H, s), 1.32 (IH, m), 1.39 (IH, m), 1.40 (IH, m), 1.54 (3H, m), 1.72 (IH, m), 1.76 (3H, s), 1.97 (IH, br s), 2.11 (3H, s), 2.52 (2H, m), 2.54 (IH, m), 2.64 (IH, dd, J= 14.9, 3.8 Hz), 2.80 (IH, dd, J= 5.0, 2.3 Hz), 2.95 (IH, dd, J = 8.6, 2.3 Hz), 3.34 (IH, m), 3.53 (IH, d, J= 10.9 Hz), 3.77 (IH, m), 5.10 (IH, d, J = 9.2 Hz), 5.17 (IH, d, J= 10.7 Hz), 5.63 (IH, dd, J= 15.2, 9.3 Hz), 5.69 (2H, m), 6.10 (IH, d, J = 10.8 Hz), 6.33 (IH, dd, J= 15.1, 10.8 Hz); (+)-HR-ESIMS m/z 589.3392 [M+Naf

Scheme 5 Procedure for Syntheses of N831 (A) Rn Carbamates

N831 R₁₇ carbamates 8-83% To a solution of compound GM137 (499 mg, 0.73 mmol), DMAP (20 mg, 0.16 mmol) and triethylamine (0.21 mL, 1.47 mmol) in anhydrous CH₂Cl₂ (12 mL) was added 4- nitroplienyl chloroformate (296 mg, 1.47 mmol). The reaction mixture was stirred at room temperature for 1 hour. Additional 4-nitrophenyl chloroformate (444 mg, 2.2 mmol) and triethylamine (0.3 mL, 2.2 mmol) were added. The mixture was stirred at room temperature for another hour, and then washed with saturated aqueous NaHCO₃ (3 x 10 mL). The organic layer was dried with anhydrous MgSO₄, filtered, and then concentrated. To a solution of crude carbonate (0.2 mmol) in anhydrous THF (5 mL) was added the respective amine (0.5-0.6 mmol). The reaction mixture was stirred at room temperature for 2 hours and then concentrated. The crude product was purified by silica gel column chromatography (gradient elution with ethyl acetate-hexane, then chloroform-methanol) to give compound 3. Deprotection of compound 3 (0.1-0.2 mmol) was carried out in methanol (4 mL) and pyridinium p-toluenesulfonate (20-25 mg). The reaction mixture was stirred at room temperature for 2 to 18 hours and then concentrated. The residue was taken into CH₂Cl₂ (5 mL) and washed with saturated aqueous NaHCO₃ (3 mL). The organic layer was dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by preparative TLC (ethyl acetate-hexane or chloroform-methanol) to give the corresponding C17-carbamate analogue.

GM155

¹H NMR (500 MHz, CDCl₃) <50.87 (3H, t, J= 7.5 Hz), 0.88 (3H, d, J = 6.9 Hz), 0.89 (3H, d, J= 7.2 Hz), 1.09 (3H, d, J= 6.7 Hz), 1.22 (3H, s), 1.29 (IH, m), 1.36 (IH, m), 1.38 (IH, m), 1.46 (IH, m), 1.57 (IH, m), 1.65 (IH, m), 1.71 (IH, m), 1.73 (3H, s), 2.10 (3H, s), 2.51 (IH, m), 2.55 (IH, dd, J= 15.0, 2.7 Hz), 2.62 (IH, dd, J= 15.0, 3.5 Hz), 2.68 (IH, m), 2.83 (2H, m), 3.17 (IH, ddd, J= 6.1, 6.1, 4.1 Hz), 3.39 (3H, s), 3.76 (IH, m), 4.41 (2H, br dd, J = 5.5, 5.5 Hz), 4.53 (IH, dd, J= 7.3, 6.7 Hz), 5.09 (H-7, IH, d, J= 9.2 Hz), 5.16 (H-Il, IH, d, J= 10.7 Hz), 5.20 (IH, br t, J= 5.5 Hz), 5.61 (IH, dd, J= 15.2, 9.4 Hz), 5.68 (IH, dd, J = 15.2, 9.2 Hz), 5.71 (IH, dd₅ J= 14.7, 6.7 Hz), 6.09 (IH, d, J= 10.8 Hz), 6.32 (IH, dd, J= 14.7, 10.8 Hz), 7.31 (IH, m), 7.70 (1-H, d, J= 7.7 Hz), 8.57 (2H, m); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.3, 12.8, 17.4, 17.7, 22.3, 24.6, 25.6, 30.8, 36.2, 39.3, 40.1, 41.1, 41.7, 43.6, 59.1, 59.3, 60.6, 70.2, 74.4, 78.7, 79.9, 83.6, 84.6, 124.8, 126.6, 127.6, 131.8, 132.9, 135.3, 136.8, 137.0, 141.4, 149.5, 157.0, 170.6, 173.0; (+)-HR-ESIMS m/z 701.4007 [M+H]⁺

GM156

¹H NMR (500 MHz, CDCl₃) (50.88 (3H, t, J= 7.5 Hz), 0.89 (3H, d, J= 7.1 Hz), 0.89 (H, d, J = 6.8 Hz), 1.11 (3H, d, J= 6.8 Hz), 1.14 (2H, m), 1.17 (2H, m), 1.22 (3H, s), 1.31 (IH, m), 1.34 (IH, m), 1.35 (IH, m), 1.38 (IH, m), 1.45 (IH, m), 1.54 (IH, m), 1.60 (2H, m), 1.64 (IH, m), 1.69 (IH, m), 1.71 (IH, m), 1.74 (3H, s), 1.93 (2H, m), 2.09 (3H, s), 2.52 (IH, m), 2.55 (IH, dd, J= 15.1, 3.0 Hz), 2.61 (IH, dd, J= 15.1, 3.6 Hz), 2.67 (IH, m), 2.84 (IH, m), 2.86 (IH, m), 3.18 (IH, ddd, J= 6.4, 6.4, 4.1 Hz), 3.39 (3H, s), 3.48 (IH, m), 3.75 (IH, m), 4.52 (br dd, J= 6.9, 6.5 Hz), 4.66 (IH, d, J= 7.9 Hz), 5.09 (IH₅ d, J = 9.2 Hz), 5.16 (IH, d, J= 10.7 Hz), 5.62 (IH, dd, J = 15.1, 9.4 Hz), 5.65 (IH, dd, J= 15.1, 9.2 Hz), 5.73 (IH, dd, J= 15.0, 8.3 Hz), 6.09 (IH, d, J= 10.7 Hz), 6.30 (IH, dd, J= 15.0, 10.7 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.8, 17.4, 17.7, 22.3, 24.7, 25.6, 25.7, 26.5, 30.8, 34.2, 36.2, 39.3, 40.1, 41.1, 41.7, 50.8, 59.1, 59.3, 60.3, 70.1, 74.4, 77.2, 79.9, 83.6, 84.4, 126.5, 127.4, 131.7, 131.9, 137.5, 141.5, 156.0, 170.6, 173.0; (+)-HR-ESIMS m/z 692.4295 [M+H]⁺ GM180

¹H NMR (500 MHz, CDCl₃) <50.87 (3H, t, J= 7.6 Hz), 0.88 (3H, d, J= 7.5 Hz), 0.89 (3H, d, J= 6.7 Hz), 1.11 (3H, d, J= 6.8 Hz), 1.22 (3H, s), 1.29 (IH, m), 1.33 (IH, m), 1.37 (IH, m), 1.45 (IH, m), 1.55 (IH, m), 1.65 (IH, m), 1.71 (IH, m), 1.74 (3H, s), 2.10 (3H, s), 2.28 (6H s), 2.46 (2H, m), 2.51 (IH, m), 2.52 (IH, dd, J= 15.0, 2.4 Hz), 2.64 (IH, dd, J= 15.0, 3.5 Hz), 2.65 (IH, m), 2.83 (IH, br d, J= 6.3 Hz), 2.86 (IH, dd, J= 8.2, 2.1 Hz), 3.18 (IH, ddd, J= 6.4, 6.4, 4.1 Hz), 3.30 (2H, m), 3.39 (3H, s), 3.76 (IH, m), 4.49 (dd, J= 7.0, 6.3 Hz), 5.09 (IH, d, J = 9.2 Hz), 5.16 (IH, d, J = 10.7 Hz), 5.43 (IH, br t, J = 5.6 Hz), 5.62 (IH, dd, J= 15.2, 4.4 Hz), 5.68 (IH, dd, J= 15.2, 9.2 Hz), 5.72 (IH, dd, J= 15.1, 8.3 Hz), 6.09 (IH, d, J= 10.9 Hz), 6.32 (IH, dd, J= 15.1, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.3, 12.8, 17.4, 17.7, 22.2, 24.6, 25.6, 30.8, 36.2, 39.2, 39.3, 40.1, 41.2, 41.7, 46.0, 59.1, 59.5, 60.5, 70.1, 74.4, 78.0, 79.9, 83.6, 84.4, 126.6, 127.3, 131.9, 132.6, 137.4, 141.5, 157.0, 170.6, 173.0; (+)-HR-ESIMS m/z 681.4213 [M+H]⁺

GM181

t, J= 7.5 Hz), 0.88 (3 H, d, J= 7.1 Hz), 0.88 (3H, d, J= 6.9 Hz), 1.10 (3H, d, J= 6.8 Hz), 1.21 (3H, s), 1.29 (IH, m), 1.35 (IH, m), 1.41 (IH, m), 1.46 (3H, m), 1.57 (IH, m), 1.63 (4H, m), 1.67 (IH, m), 1.70 (IH, m), 1.74 (3H, s), 2.09 (3H, s), 2.48 (4H, m), 2.49 (2H, m), 2.50 (IH, m), 2.55 (IH, dd, J= 15.1, 2.8 Hz), 2.62 (IH, dd, J = 15.1, 3.4 Hz), 2.66 (IH, m), 2.84 (IH, br d, J = 6.2 Hz), 2.86 (IH, dd, J= 8.1, 2.1 Hz), 3.17 (IH, ddd, J = 6.4, 6.4, 4.1 Hz), 3.32 (2H, m), 3.39 (3H, s), 3.75 (IH, m), 4.50 (IH, dd, J = 7.1, 6.2 Hz), 5.09 (IH, d, J= 9.2 Hz), 5.16 (IH, d, J= 10.7 Hz), 5.55 (IH, br s), 5.61 (IH, dd, J = 15.1, 9.3 Hz), 5.67 (IH, dd, J= 15.1, 9.2 Hz), 5.71 (IH, dd, J= 15.1, 8.4 Hz), 6.09 (IH, d, J = 10.9 Hz), 6.30 (IH, d, J = 15.1, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.3, 12.8, 17.4, 17.8, 22.3, 24.6, 25.0, 25.6, 26.4, 30.8, 36.2, 38.5, 39.3, , 40.1, 41.2, 41.7, 55.3, 58.5, 59.0, 59.5, 60.5, 70.1, 74.4, 77.9, 79.9, 83.6, 84.4, 126.5, 127.4, 131.9, 132.6, 137.4, 141.5, 157.0, 170.6, 173.0; (+)-HR-ESIMS m/z 721.4510 [M+H]⁺

GM182

¹H NMR (500 MHz, CDCl₃) £0.85 (3H, t, J = 8.0 Hz), 0.86 (6H, m), 1.11 (3H, br s), 1.20 (3H, s), 1.31 (IH, m), 1.34 (IH, m), 1.37 (IH, m), 1.44 (IH, m), 1.53 (IH, m), 1.63 (IH, m), 1.69 (2H, m), 1.72 (3H, s), 2.07 (IH, m), 2.09 (3H, s), 2.51 (IH, m), 2.53 (IH, dd, J = 14.9, 2.6 Hz), 2.59 (IH, dd, J= 14.9, 2.9 Hz), 2.68 (IH, m), 2.83 (2H, br d, J = 7.3 Hz), 3.15 (2H, m), 3.38 (3H, s), 3.46 (IH, m), 3.56 (IH, m), 3.59 (2H, m), 3.74 (IH, m), 4.48 (IH, dd, J= 7.3, 6.8 Hz), 5.07 (IH, d, J= 9.3 Hz), 5.13 (IH, d, J= 10.6 Hz), 5.61 (IH, dd, J = 15.1, 9.4 Hz), 5.65 (IH, dd, J= 15.1, 9.3 Hz), 5.75 (IH, dd, J= 15.0, 7.4 Hz), 6.09 (IH, d, J= 10.6 Hz), 6.30 (IH, dd, J= 15.0, 10.6 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.3, 12.8, 17.5, 17.6, 22.2, 24.7, 25.6, 30.8, 35.2, 36.2, 39.3, 40.2, 41.0, 41.7, 45.6, 52.3, 55.1, 59.1, 59.4, 60.6, 70.1, 74.3, 78.7, 79.9, 83.6, 84.4, 126.6, 127.2, 131.9, 132.5, 137.4, 141.4, 155.3, 170.6, 173.0; (+)-HR-ESIMS m/z 679.4012 [M+H]⁺

GM216

¹H NMR (500 MHz, CDCl₃) 50.84 (3H, t, J = 7.5 Hz), 0.86 (3H, d, J = 6.7 Hz), 0.88 (3 H, d, J = 7.1 Hz)₅ 1.10 (3H, d, J= 6.8 Hz), 1.11 (2H, m), 1.22 (3H, s), 1.22 (IH, m), 1.23 (2H, m), 1.30 (IH, m), 1.35 (IH, m), 1.36 (IH, m), 1.44 (IH, m), 1.54 (IH, m), 1.63 (2H, m), 1.64 (IH, m), 1.70 (IH, m), 1.73 (3H, s), 1.80 (2H, m), 1.84 (IH, m), 2.09 (3H, s), 2.28 (IH, m), 2.51 (IH, m), 2.53 (5H, m), 2.61 (IH, dd, J = 15.1, 3.4 Hz), 2.68 (IH, ra), 2.83 (IH, m), 2.85 (IH, m), 3.17 (IH, ddd, J= 6.4, 6.4, 4.1 Hz), 3.39 (3H, s), 3.49 (4H, m), 3.74 (IH, m), 4.53 (IH, dd, J= 7.1, 6.8 Hz), 5.09 (IH, d, J= 9.2 Hz), 5.15 (IH, d, J= 10.1 Hz), 5.61 (IH, dd, J= 15.1, 9.2 Hz), 5.65 (IH, dd, J= 15.1, 9.4 Hz), 5.72 (IH, dd, J= 15.1, 8.3 Hz), 6.09 (IH, d, J = 10.8 Hz), 6.30 (IH, dd, J = 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) 5 10.9, 11.3, 13.1, 17.7, 17.9, 22.5, 25.2, 25.9, 27.2, 27.5, 30.2, 31.2, 36.6, 39.6, 40.6, 41.2, 41.9, 45.5, 50.1, 59.4, 59.5, 60.7, 65.0, 70.5, 74.4, 78.9, 80.1, 83.8, 84.9, 126.8, 127.5, 131.9, 132.1, 137.8, 141.7, 155.5, 170.6, 173.0; (+)-HR-ESIMS m/z 761.5118 [M+H]⁺ GM230

¹H NMR (500 MHz, CDCl₃) 50.88 (3H, t, J= 7.6 Hz), 0.89 (3 H, d, J= 6.7 Hz), 0.89 (3H, d, J= 6.7 Hz), 1.11 (3H, d, J= 6.8 Hz), 1.22 (3H, s), 1.27 (IH, m), 1.34 (IH, m), 1.37 (IH, m), 1.45 (IH, m), 1.55 (IH, m), 1.65 (IH, m), 1.68 (IH, m), 1.75 (3H, s), 1.81 (4H, m), 2.10 (3H, s), 2.51 (IH, m), 2.56 (IH, m), 2.58 (4H, m), 2.63 (IH, m), 2.65 (3H, m), 2.84 (IH, m), 2.86 (IH, dd, J= 8.2, 2.0 Hz), 3.18 (IH, ddd, J= 6.3, 6.3, 4.2 Hz), 3.33 (2H, m), 3.40 (3H, s), 3.76 (IH, m), 4.51 (IH, dd, J= 6.8, 6.2 Hz), 5.02 (IH, d, J= 9.2 Hz), 5.16 (IH, d, J= 10.6 Hz), 5.43 (IH, br s), 5.62 (IH, dd, J = 15.2, 9.4 Hz), 5.67 (IH, dd, J = 15.2, 9.2 Hz), 5.73 (IH, dd, J= 15.0, 8.3 Hz), 6.10 (IH, d, J= 10.8 Hz), 6.32 (IH, dd, J= 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.3, 12.8, 17.4, 17.7, 22.2, 24.4, 24.6, 25.6, 30.8, 36.2, 39.3, 40.1, 40.4, 41.2, 41.7, 54.9, 55.9, 59.1, 59.4, 60.5, 70.1, 74.4, 77.9, 79.9, 83.5, 84.4, 126.5, 127.4, 131.9, 132.6, 137.4, 141.4, 157.0, 170.6, 173.0; (+)-HR-ESIMS m/z 707.4481 [M+H]⁺ GM231

¹H NMR (500 MHz, CDCl₃) 50.85 (3H, t, J = IA Hz), 0.86 (3H, d, J= 7.5 Hz), 0.87 (3 H, d, J = 7.1 Hz), 1.15 (3H, d, J= 6.8 Hz), 1.20 (3H, s), 1.30 (IH, m), 1.35 (IH, m), 1.36 (IH, m), 1.45 (IH, m), 1.55 (IH, m), 1.63 (IH, m), 1.70 (IH, m), 1.76 (3H, s), 2.09 (3H, s), 2.45 (2H, m), 2.48 (4H, m), 2.51 (IH, m), 2.54 (IH, dd, J = 15.0, 2.4 Hz), 2.62 (IH, dd, J = 15.0, 3.6 Hz), 2.67 (IH, m), 2.85 (IH, m) 2.86 (IH, m), 3.17 (IH, ddd, J = 6.4, 6.4, 4.1 Hz), 3.30 (2H, m), 3.38 (3H, s), 3.71 (4H, m), 3.75 (IH, m), 4.51 (IH, br dd, J= 7.3, 6.4 Hz), 5.08 (IH, d, J = 9.2 Hz), 5.16 (IH, d, J = 10.7 Hz), 5.27 (IH, br t, J = 5.5 Hz), 5.63 (IH, dd, J= 15.1, 9.4 Hz), 5.67 (IH, dd, J= 15.1, 9.2 Hz), 5.74 (IH, dd, J= 15.0, 8.3 Hz), 6.08 (IH, d, J= 10.5 Hz), 6.32 (IH, dd, J= 15.0, 10.5 Hz); ¹³C NMR (125 MHz, CDCl₃) S 10.9, 11.2, 12.8, 17.4, 17.5, 22.2, 24.7, 25.6, 30.8, 36.2, 38.3, 39.3, 40.1, 41.5, 41.7, 54.3, 58.4, 59.0, 59.4, 60.5, 67.9, 70.1, 74.4, 77.8, 79.8, 83.5, 84.4, 126.6, 127.4, 131.9, 132.7, 137.3, 141.4, 156.9, 170.6, 173.0; (+)-HR-ESIMS m/z 723.4277 [M+H]⁺

GM232

¹H NMR (500 MHz, CDCl₃) 50.86 (3H, t, J= 7.6 Hz), 0.87 (3H, d, J= 7.4 Hz), 0.88 (3H, d, J= 6.7 Hz), 1.10 (3H, d, J= 6.8 Hz), 1.21 (3H, s), 1.32 (IH, m), 1.35 (IH, m), 1.38 (IH, m), 1.46 (IH, m), 1.54 (IH, m), 1.64 (IH, m), 1.69 (2H, m), 1.70 (IH, m), 1.74 (3H, s), 2.09 (3H, s), 2.43 (2H, m), 2.45 (4H, m), 2.51 (IH, m), 2.53 (IH, dd, J= 15.1, 2.7 Hz), 2.62 (IH, dd, J= 15.1, 3.4 Hz), 2.64 (IH, m), 2.82 (IH, br d, J= 6.4 Hz), 2.83 (IH, dd, J= 8.2, 2.0 Hz), 3.16 (IH, ddd, J= 6.4, 6.4, 4.1 Hz), 3.28 (2H, q, J = 6.0 Hz), 3.38 (3H, s), 3.71 (4H, t, J = 4.6 Hz), 3.75 (IH, m), 4.48 (IH, dd, J = 7.0, 6.4 Hz), 5.09 (IH, d, J = 9.2 Hz), 5.15 (IH, d, J= 10.7 Hz), 5.62 (IH, dd, J= 15.3, 9.4 Hz), 5.65 (IH, dd, J = 15.3, 9.2 Hz), 5.71 (IH, dd, J= 15.1, 8.5 Hz), 5.72 (IH, br s), 6.08 (IH, d, J= 10.8 Hz), 6.30 (IH, dd, J= 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.3, 12.8, 17.4, 17.7, 22.2, 24.6, 25.6, 26.7,

30.8, 36.2, 39.3, 40.1, 41.2, 41.4, 41.7, 54.6, 58.1, 59.0, 59.5, 60.5, 67.9, 70.1, 74.4, 77.8,

79.9, 83.5, 84.4, 126.6, 127.3, 131.9, 132.6, 137.4, 141.4, 157.0, 170.6, 173.0; (+)-HR- ESIMS m/z 737.4397 [M+H]⁺

GM234

¹H NMR (500 MHz, CDCl₃) £0.88 (3H, t, J = 7.5 Hz), 0.89 (3H, d, J= 6.6 Hz), 0.90 (3 H, d, J= 7.0 Hz), 1.12 (3H, d, J= 6.9 Hz), 1.22 (3H, s), 1.31 (IH, m), 1.35 (IH, m), 1.38 (IH, m), 1.45 (IH, m), 1.55 (IH, m), 1.66 (IH, m), 1.70 (IH, m), 1.75 (3H, s), 2.10 (3H, s), 2.33 (3H, s), 2.40 (4H, m), 2.51 (IH, m), 2.55 (IH, dd, J= 14.9, 2.9 Hz), 2.63 (IH, dd, J= 14.9, 3.7 Hz), 2.69 (IH, m), 2.86 (2H, m), 3.19 (IH, ddd, J= 6.4, 6.4, 4.0 Hz), 3.40 (3H, s), 3.54 (4H, m), 3.76 (IH, m), 4.53 (IH, dd, J= 7.8, 6.9 Hz), 5.09 (IH, d, J= 9.2 Hz), 5.17 (IH, d, J= 10.7 Hz), 5.62 (IH, dd, J= 15.2, 9.4 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.73 (IH, dd, J = 14.9, 8.2 Hz), 6.10 (IH, d, J = 10.8 Hz), 6.31 (IH, dd, J= 14.9, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.8, 17.4, 17.5, 22.2, 24.6, 25.6, 30.8, 36.2, 39.3, 40.1, 41.0, 41.7, 44.6, 47.0, 55.6, 59.1, 59.2, 60.5, 70.1, 74.3, 78.8, 79.8, 83.5, 84.4, 126.6, 127.4, 131.9, 132.6, 137.2, 141.4, 155.5, 170.6, 173.0. (+)-HR-ESIMS m/z 693.4326 [M+H]⁺

GM235

¹H NMR (500 MHz, CDCl₃) £0.88 (3H, t, J= 7.5 Hz), 0.89 (3H, d, J= 7.1 Hz), 0.89 (3H, d, J= 6.7 Hz), 1.11 (3H, d, J= 6.8 Hz), 1.22 (3H, s), 1.30 (IH, m), 1.34 (IH, m), 1.38 (IH, m), 1.45 (IH, m), 1.55 (IH, m), 1.65 (IH, m), 1.70 (3H, m), 1.75 (3H, s), 2.11 (3H, s), 2.26 (6H, s), 2.39 (2H, m), 2.52 (IH, m), 2.55 (IH, dd, J= 15.1, 2.9 Hz), 2.63 (IH, dd, J= 15.1, 3.6 Hz), 2.66 (IH, m), 2.84 (IH, m), 2.86 (IH, m), 3.18 (IH, ddd, J = 6.4, 6.4, 4.1 Hz), 3.28 (2H, br q, J = 5.8 Hz), 3.40 (3H, s), 3.75 (IH, m), 4.51 (IH, dd, J= 7.3, 6.3 Hz), 5.09 (IH, d, J= 9.2 Hz), 5.17 (IH, d, J = 10.7 Hz), 5.62 (IH, dd, J= 15.1, 9.4 Hz), 5.63 (IH, br s), 5.67 (IH, dd, J= 15.1, 9.2 Hz), 5.73 (IH, dd, J= 15.0, 8.3 Hz), 6.10 (IH, d, J= 10.8 Hz), 6.32 (IH, dd, J= 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) £ 10.9, 11.2, 12.7, 17.4, 17.7, 22.2, 24.6, 25.5, 27.9, 30.7, 36.2, 39.3, 40.1, 41.1, 41.6, 46.2, 58.7, 59.0, 59.4, 60.4, 70.1, 74.3, 77.6, 79.8, 83.5, 84.4, 126.5, 127.3, 131.9, 132.5, 137.4, 141.4, 157.0, 170.6, 172.9; (+)-HR-ESIMS m/z 695.4355 [M+H]⁺

GM236

¹H NMR (500 MHz, CDCl₃) £0.88 (3H, t, J= 7.0 Hz), 0.89 (3H, d, J= 7.0 Hz), 0.89 (3H, d, J= 7.0 Hz), 1.11 (3H, d, J= 6.8 Hz), 1.22 (3 H, s), 1.28 (IH, m), 1.33 (IH, m), 1.38 (IH, m), 1.45 (IH, m), 1.55 (IH, m), 1.65 (IH, m), 1.69 (IH, m), 1.74 (3H, s), 2.10 (3H, s), 2.51 (IH, m), 2.55 (IH, dd, J= 15.0, 2.3 Hz), 2.64 (IH, dd, J= 15.0, 3.8 Hz), 2.69 (IH, m), 2.85 (IH, m), 2.86 (IH, m), 2.94 (6H, s), 3.18 (IH, m), 3.40 (3H, s), 3.53 (IH, d, J= 10.9 Hz), 3.76 (IH, m), 4.49 (IH, dd, J= 7.1, 6.9 Hz), 5.09 (IH, d, J= 9.1 Hz), 5.16 (IH, d, J= 10.7 Hz), 5.62 (IH, dd, J= 15.2, 9.4 Hz), 5.68 (IH, dd, J= 15.2, 9.1 Hz), 5.74 (IH, dd, J= 15.1, 8.2 Hz), 6.10 (IH, d, J= 10.9 Hz), 6.32 (IH, dd, J= 15.1, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.5, 22.2, 24.7, 25.5, 30.7, 36.2, 36.7, 37.5, 39.3, 40.2, 41.0, 41.6, 59.1, 59.4, 60.5, 70.1, 74.3, 78.8, 79.8, 83.5, 84.4, 126.5, 127.2, 131.9, 132.5, 137.4, 141.4, 156.8, 170.6, 173.0; (+)-HR-ESIMS m/z 660.3640 [M+Naf GM237

¹H NMR (500 MHz, CDCl₃) 50.88 (3H, t, J= 7.6 Hz), 0.89 (3H, d, J= 6.5 Hz), 0.90 (3H, d, J= 7.0 Hz), 1.13 (3H, d, J= 6.9 Hz), 1.23 (3H, s), 1.30 (IH, m), 1.36 (2H, m), 1.45 (IH, m), 1.55 (IH, m), 1.66 (IH, m), 1.71 (IH, m), 1.74 (3H, s), 2.11 (3H, s), 2.50 (IH, m), 2.54 (IH, dd, J= 15.0, 2.9 Hz), 2.62 (IH, dd, J= 15.0, 3.8 Hz), 2.73 (IH, m), 2.87 (IH, m), 2.89 (IH, m), 3.19 (IH, ddd, J= 6.4, 6.4, 4.0 Hz), 3.39 (3H, s), 3.56 (4H, m), 3.63 (4H, m), 3.76 (IH, m), 4.56 (IH, dd, J= 7.1, 7.0 Hz), 5.10 (IH, d, J= 9.2 Hz), 5.17 (IH, d, J= 10.7 Hz), 5.62 (IH, dd, J= 15.2, 9.4 Hz), 5.68 (IH, dd, J= 15.2, 9.2 Hz), 5.74 (IH, dd, J= 15.1, 8.2 Hz), 6.11 (IH, d, J= 10.9 Hz), 6.33 (IH, dd, J= 15.1, 10.9 Hz), 6.67 (IH, m), 6.67 (IH, m), 7.52 (IH, t, J = 7.2 Hz), 8.21 (IH, m); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.6, 22.2, 24.6, 25.6, 30.8, 36.2, 39.3, 40.1, 41.0, 41.6, 44.5, 46.1, 59.1, 59.2, 60.5, 70.1, 74.3, 79.1, 79.8, 83.5, 84.3, 108.3, 114.7, 126.6, 127.4, 131.8, 132.7, 137.2, 138.8, 141.4, 148.7, 155.7, 159.9, 170.6, 173.0; (+)-HR-ESIMS m/z 756.4329 [M+H]⁺

GM244

¹H NMR (500 MHz, CDCl₃) £0.84 (3H, t, J= 7.5 Hz), 0.85 (3H, d, J= 7.1 Hz), 0.86 (3H, d, J= 6.5 Hz), 1.08 (3H, d, J= 6.8 Hz), 1.19 (3H, s), 1.26 (IH, m), 1.33 (IH, m), 1.38 (IH, m), 1.44 (IH, m), 1.52 (IH, br dt, J= 12.2, 2.5 Hz), 1.62 (IH, m), 1.67 (IH, m), 1.71 (3H, s), 2.07 (3H, s), 2.28 (3H, s), 2.45 - 2.50 (HH, m), 2.52 (IH, dd, J = 15.0, 2.8 Hz), 2.59 (IH, dd, J= 15.0, 3.5 Hz), 2.65 (IH, m), 2.82 (IH, br d, J= 6.2 Hz), 2.84 (IH, dd, J= 8.1, 2.0 Hz), 3.14 (IH, ddd, J= 6.3, 6.3, 4.1 Hz), 3.26 (2H, m), 3.36 (3H, s), 3.72 (IH, m), 4.48 (IH₅ dd J= 7.3, 6.2 Hz), 5.06 (IH, d, J= 9.3 Hz), 5.13 (IH, d, J= 10.6 Hz), 5.27 (IH, br t, J= 5.2 Hz), 5.58 (IH, dd, J= 15.3, 9.5 Hz), 5.65 (IH, dd, J= 15.3, 9.3 Hz), 5.70 (IH, dd, J = 15.0, 8.3 Hz), 6.07 (IH, d, J= 10.7 Hz), 6.29 (IH, dd, J= 15.0, 10.7 Hz); ¹³C NMR (125 MHz, CDCl₃) 5 10.9, 11.2, 12.7, 17.4, 17.7, 22.2, 24.6, 25.5, 30.7, 36.2, 38.5, 39.3, 40.1, 41.1, 41.6, 46.8, 53.6, 55.9, 57.8, 59.0, 59.3, 60.4, 70.1, 74.3, 77.8, 79.8, 83.5, 84.4, 126.5, 127.3, 131.9, 132.6, 137.3, 141.4, 156.8, 170.6, 172.9; (+)-HR-ESIMS m/z 736.4883 [M+H]⁺

GM246

¹H NMR (500 MHz, CDCl₃) <50.83 (3H, d, J= 7.5 Hz), 0.84 (3H, d, J= 7.2 Hz), 0.85 (3H, t, J= 6.7 Hz), 1.08 (3H, d, J= 6.8 Hz), 1.19 (3H, s), 1.28 (IH, m), 1.32 (IH, m), 1.35 (IH, m), 1.43 (IH, m), 1.52 (IH, m), 1.61 (IH, m), 1.64 (2H, m), 1.67 (IH, m), 1.71 (3H, s), 2.06 (3H, s), 2.26 (3H, s), 2.40 (4H, m), 2.44 (4H, m), 2.46 (2H, m), 2.50 (IH, m), 2.51 (IH, dd, J= 15.1, 2.9 Hz), 2.59 (IH, dd, J= 15.1, 3.6 Hz), 2.63 (IH, m), 2.81 (IH, br d, J= 7.2 Hz), 2.95 (IH, dd, J = 8.2, 1.9 Hz), 3.13 (IH, ddd, J= 6.4, 6.4, 4.1 Hz), 3.24 (2H, m), 3.36 (3H, s), 3.72 (IH, m), 4.47 (IH, br dd, J= 7.2, 6.6 Hz), 5.05 (IH, d, J= 9.2 Hz), 5.12 (IH, d, J= 10.7 Hz), 5.61 (IH, dd, J= 15.1, 9.4 Hz), 5.65 (IH, dd, J= 15.1, 9.2 Hz), 5.71 (IH, dd, J= 15.0, 8.3 Hz), 5.81 ( IH, br t, J= 5.8 Hz), 6.06 (IH, d, J= 10.8 Hz), 6.28 (IH, dd, J = 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.3, 12.7, 17.4, 17.6, 22.2, 24.6, 25.5, 27.0, 30.7, 36.2, 39.3, 40.1, 41.1, 41.5, 41.7, 46.9, 54.0, 56.0, 57.7, 59.0, 59.4, 60.3, 70.1, 74.3, 77.6, 79.8, 83.5, 84.4, 126.5, 127.2, 131.9, 132.6, 137.5, 141.4, 156.9, 170.6, 172.9; (+)-HR-ESIMS m/z 750.4894 [M+H]⁺ GM250

¹H NMR (500 MHz, CDCl₃) 50.81 (3H, t, J = 7.6 Hz), 0.85 (3H, d, J= 7.4 Hz), 0.86 (3H, d, J= 6.7 Hz), 1.07 (3H, d, J= 6.8 Hz), 1.18 (3H, s), 1.26 (IH, m), 1.31 (IH, m), 1.35 (IH, m), 1.42 (IH, m), 1.51 (IH, br t, J = 12.2 Hz), 1.61 (IH, m), 1.65 (IH, m), 1.70 (3H, s), 1.72, (2H, m), 1.79 (4H, m), 2.07 (3H, s), 2.45-2.65 (1OH, m), 2.79 (IH₅ br d, J= 6.1 Hz), 2.82 (IH, dd, J= 8.0, 1.9 Hz), 3.14 (IH, ddd, J= 6.3, 6.3, 4.2 Hz), 3.25, (2H, m), 3.36 (3H, s), 3.73 (IH, m), 4.49 (IH, dd, J= 6.9, 6.1 Hz), 5.06 (IH, d, J = 9.3 Hz), 5.12 (IH, d, J = 10.6 Hz), 5.58, (IH, dd, J= 15.2, 9.4 Hz), 5.63 (IH, dd, J= 15.2, 9.3 Hz), 5.68, (IH, dd, J= 15.0, 8.5 Hz), 5.70 (IH, m), 6.06 (IH, d, J= 10.8 Hz). 6.28 (IH, dd, J= 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.8, 11.3, 12.7, 17.4, 17.6, 22.1, 24.3, 24.6, 25.5, 29.0, 30.6, 36.2, 39.3, 40.1, 41.0, 41.1, 41.6, 55.0, 55.2, 59.0, 59.4, 60.4, 70.1, 74.3, 77.7, 79.8, 83.5, 84.4, 126.5, 127.3, 131.9, 132.6, 137.4, 141.4, 157.0, 170.6, 173.0; (+)-HR-ESIMS m/z 721.4308 [M+H]⁺

GM251

¹H NMR (500 MHz, CDCl₃) £0.87 (3H, t, J= 7.2 Hz), 0.87 (3H, d, J= 7.0 Hz), 0.88 (3H, d, J= 6.5 Hz), 1.09 (3H, d, J= 6.7 Hz), 1.21 (3H, s), 1.30 (IH, m), 1.33 (IH, m), 1.36 (IH, m), 1.44 (IH, m), 1.52 (2H, m), 1.54 (IH, m), 1.64 (IH, m), 1.68 (IH, m), 1.73 (4H, m), 1.74 (3H, s), 1.81 (2H, m), 2.09 (3H, s), 2.48 (IH, m), 2.53 (IH, m), 2.55 (6H, m), 2.62

(IH, m), 2.64 (IH, m), 2.82 (IH, m), 2.83 (IH, m), 3.16 (IH, m), 3.29 (2H, m), 3.38 (3H, s), 3.74 (IH, m), 4.46 (IH, dd, J= 6.8, 6.6 Hz), 5.08 (IH, d, J = 92 Hz), 5.15 (IH, d, J = 10.6 Hz), 5.61 (IH, dd, J= 15.2, 9.4 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.70 (IH, dd, J = 15.0, 8.3 Hz), 6.03 (IH, br s), 6.08 (IH, d, J = 10.9 Hz), 6.31 (IH, dd, J= 15.0, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.8, 11.3, 12.7, 17.3, 17.6, 22.2, 24.56, 24.64, 25.5, 25.9, 26.2, 30.7, 36.2, 39.3, 40.1, 40.9, 41.1, 41.6, 55.1, 57.7, 59.0, 59.5, 60.4, 70.1, 74.3, 77.9, 79.8, 83.5, 84.4, 126.5, 127.2, 131.8, 132.5, 137.4, 141.3, 157.1, 170.5, 172.9; (+)-HR- ESIMS m/z 735.4570 [M+H]⁺

GM257

¹H NMR (500 MHz, CDCl₃) <50.87 (3H, t, J= 7.5 Hz), 0.88 (3H, d, J= 6.7 Hz), 0.88 (3H, d, J= 7.0 Hz), 1.11 (3H, d, J= 6.9 Hz), 1.21 (3H, s), 1.27 (3H, t, J= 6.3 Hz), 1.29 (IH, m), 1.33 (IH, m), 1.38 (IH, m), 1.43 (IH, m), 1.54 (IH, m), 1.65 (IH, m), 1.70 (IH, m), 1.73 (3H, s), 2.10 (3H, s), 2.50 (IH, m), 2.53 (IH, dd, J= 15.0, 3.0 Hz), 2.62 (IH, dd, J= 15.0, 3.8 Hz), 2.70 (IH, m), 2.85 (2H, br d, J= 7.2 Hz), 3.17 (IH, ddd, J= 6.5, 6.5, 3.9 Hz), 3.39 (3H, s), 3.48 (8H, m), 3.75 (IH, m), 4.17 (2H, q, J= 6.3 Hz), 4.52 (IH, dd, J= 7.2, 6.8 Hz), 5.08 (IH, d, J = 9.3 Hz), 5.15 (IH, d, J= 10.7 Hz), 5.61 (IH, dd, J = 15.2, 9.4 Hz), 5.67 (IH, dd, J= 15.2, 9.3 Hz), 5.71 (IH, dd, J= 15.1, 8.2 Hz), 6.09 (IH, d, J= 10.9 Hz), 6.31 (IH, dd, J = 15.1, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.1, 12.7, 15.5, 17.3,

17.5, 22.2, 24.5, 25.5, 30.6, 30.7, 36.2, 39.2, 40.0, 41.0, 41.6, 44.4, 44.6, 59.0, 59.1, 60.5,

62.6, 70.1, 74.3, 79.2, 79.8, 83.5, 84.3, 126.6, 127.4, 131.7, 132.8, 137.0, 141.3, 155.6, 156.3, 170.5, 172.; (÷)-HR-ESIMS m/z 773.4069 [M+Naf GM258

¹H NMR (500 MHz, CDCl₃) £0.87 (3H, t, J= 7.6 Hz), 0.88 (3H, d, J= 7.4 Hz), 0.89 (3H, d, J = 7.0 Hz), 1.11 (3H, d, J= 6.9 Hz), 1.21 (3H, s), 1.29 (IH, m), 1.34 (IH, m), 1.39 (IH, m), 1.43 (IH, m), 1.54 (IH, m), 1.65 (IH, m), 1.69 (IH, m), 1.74 (3H, s), 2.09 (3H, s), 2.50 (IH, m), 2.54 (IH, dd, J= 14.9, 2.9 Hz), 2.62 (IH, dd, J= 14.9, 3.7 Hz), 2.69 (IH, m), 2.85 (2H, m), 3.18 (IH, ddd, J= 6.4, 6.4, 3.9 Hz), 3.39 (3H, s), 3.49 (4H, m), 3.66 (4H, m), 3.75 (IH, m), 4.53 (IH, dd, J= 7.0, 6.8 Hz), 5.08 (IH, d, J= 9.2 Hz), 5.16 (IH, d, J= 10.7 Hz), 5.61 (IH, dd, J= 15.2, 9.4 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.72 (IH, dd, J= 15.1, 8.2 Hz), 6.09 (IH, d, J = 10.8 Hz), 6.30 (IH, dd, J = 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.5, 22.2, 24.6, 25.5, 30.8, 36.2, 39.3, 40.1, 41.0, 41.6, 45.2, 59.1, 59.2, 60.5, 67.5, 70.1, 74.3, 79.1, 79.8, 83.5, 84.3, 126.6, 127.4, 131.8, 132.7, 137.1, 141.3, 155.7, 170.6, 173.0. (+)-HR-ESIMS m/z 702.3861 [M+Na]⁺

GM259

¹H NMR (500 MHz, CDCl₃) 50.87 (3H, t, J= 7.4 Hz), 0.88 (3H, d, J= 6.7 Hz), 0.89 (3H, d, J= 7.0 Hz), 1.11 (3H, d, J= 6.9 Hz), 1.21 (3H, s), 1.29 (IH, m), 1.33 (IH, m), 1.37 (IH, m), 1.44 (IH, m), 1.54 (IH, m), 1.64 (IH, m), 1.70 (IH, m), 1.73 (3H, s), 1.87 (4H, m), 2.10 (3H, s), 2.51 (IH, m), 2.54 (IH, dd, J= 14.9, 2.9 Hz), 2.62 (IH, dd, J= 14.9, 3.6 Hz), 2.68 (IH, m), 2.85 (2H, br d, J= 7.2 Hz), 3.18 (IH, ddd, J= 6.4, 6.4, 4.0 Hz), 3.39 (3H, s), 3.40 (4H, ni), 3.53 (IH, br d, J = 10.5 Hz), 3.75 (IH, m), 4.50 (IH, dd, J = 7.2, 6.8 Hz), 5.08 (IH, d, J= 9.2 Hz), 5.16 (IH, d, J= 10.7 Hz), 5.61 (IH, dd, J= 15.2, 9.3 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.75 (IH, dd, J= 15.1, 8.2 Hz), 6.10 (IH, d, J= 10.8 Hz), 6.31 (IH, dd, J = 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 11.0, 11.3, 12.7, 17.4, 17.6, 22.3, 24.7, 25.6, 25.9, 26.7, 30.8, 36.2, 39.3, 40.3, 41.1, 41.7, 46.7, 47.3, 59.2, 59.5, 60.6, 67.5, 70.2, 74.4, 78.4, 79.9, 83.6, 84.3, 126.6, 127.2, 132.0, 132.5, 137.6, 141.5, 155.3, 170.6, 173.0; (+)-HR-ESIMS m/z 686.3821 [M+Na]⁺

GM260

¹H NMR (500 MHz, CDCl₃) <50.85 (3H, t, J= 7.5 Hz), 0.86 (3H, d, J= 6.7 Hz), 0.87 (3H, d, J = 7.0 Hz), 1.09 (3H₅ d, J= 6.9 Hz), 1.19 (3H, s), 1.33 (IH, m), 1.35 (IH, m), 1.39 (IH, m), 1.42 (IH, m), 1.51 (4H, m), 1.55 (IH, m), 1.60 (2H, m), 1.63 (IH, m), 1.68 (IH, m), 1.71 (3H, s), 2.07 (3H, s), 2.50 (IH, m), 2.53 (IH, dd, J= 15.0, 2.9 Hz), 2.60 (IH, dd, J = 15.0, 3.6 Hz), 2.67 (IH, m), 2.82 (IH, dd, J= 8.4, 2.1 Hz), 2.84 (IH, dd, J= 6.4, 2.1 Hz), 3.16 (IH, ddd, J= 6.4, 6.4, 4.0 Hz), 3.37 (3H, s), 3.41 (4H, m), 3.73 (IH, m), 4.51 (IH, dd, J= 6.9, 6.4 Hz), 5.06 (IH, d, J= 9.3 Hz), 5.14 (IH, d, J= 10.7 Hz), 5.59 (IH, dd, J= 15.2, 9.4 Hz), 5.65 (IH, dd, J= 15.2, 9.3 Hz), 5.71 (IH, dd, J= 15.1, 8.2 Hz), 6.08 (IH, d, J = 10.8 Hz), 6.29 (IH, dd, J= 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) £ 10.9, 11.2, 12.7, 17.4, 17.5, 22.2, 24.7, 25.4, 25.5, 26.7, 30.7, 36.2, 39.3, 40.1, 41.0, 41.7, 45.9, 59.1, 59.3, 60.3, 70.1, 74.3, 78.2, 79.8, 83.5, 84.4, 126.5, 127.2, 131.9, 132.4, 137.5, 141.4, 155.6, 170.6, 173.0; (÷)-HR-ESIMS m/z 678.4315 [M+H]⁺

GM261

¹H NMR (500 MHz, CDCl₃) £0.87 (3H₅ X, J= 7.3 Hz), 0.88 (3H, d, J= 8.1 Hz), 0.90 (3H, d, J= 7.3 Hz), 1.13 (3H, d, J= 6.9 Hz), 1.22 (3H, s), 1.31 (IH, m), 1.36 (IH, m), 1.38 (IH, m), 1.44 (IH, m), 1.54 (IH, m), 1.64 (IH, m), 1.67 (IH, m), 1.73 (3H, s), 2.10 (3H, s), 2.49 (IH, m), 2.52 (IH, m), 2.61 (IH, dd, J= 15.0, 3.8 Hz), 2.71 (IH, m), 2.86 (2H, m), 3.17 (4H, m), 3.18 (IH, m), 3.39 (3H, s), 3.51 (IH, d, J= 11.0 Hz), 3.67 (4H, m), 3.75 (IH, m), 4.55 (IH, dd, J= 7.0, 6.9 Hz), 5.09 (IH, d, J= 9.2 Hz), 5.16 (IH, d, J= 10.7 Hz), 5.61 (IH, dd, J= 15.2, 9.4 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.74 (IH, dd, J= 15.1, 8.2 Hz), 6.10 (IH, d, J = 10.8 Hz), 6.32 (IH, dd, J = 15.1, 10.8 Hz), 6.91 (3H, m), 7.28 (2H, m); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.5, 22.2, 24.6, 25.5, 30.7, 36.2, 39.2, 40.1, 41.0, 41.6, 44.8, 50.4, 59.1, 59.2, 60.5, 70.1, 74.3, 79.0, 79.8, 83.5, 84.3, 117.7, 121.4, 126.6, 127.4, 130.2, 131.8, 132.7, 137.2, 141.3, 152.0, 155.6, 170.6, 173.0; (+)-HR-ESIMS m/z 777.4866 [M+Na]⁺

GM262

¹H NMR (500 MHz, CDCl₃) £0.85 (3H, t, J= 7.5 Hz), 0.86 (3H, d, J= 6.6 Hz), 0.87 (3H, d, J= 6.9 Hz), 1.10 (3H, d, J= 6.9 Hz), 1.19 (3H, s), 1.30 (IH, m), 1.35 (2H, m), 1.42 (IH, m), 1.55 (IH, m), 1.64 (IH, m), 1.68 (IH, m), 1.71 (3H, s), 2.08 (3H, s), 2.50 (IH, m), 2.52

(IH, dd, J= 14.9, 3.0 Hz), 2.60 (IH, dd, J= 14.9, 3.6 Hz), 2.71 (IH, m), 2.84 (IH, dd, J = 8.7, 2.1 Hz), 2.86 (IH, dd, J = 7.0, 2.1 Hz), 3.16 (IH, ddd, J= 6.4, 6.4, 4.0 Hz), 3.37 (3H, s), 3.56 (4H, m), 3.73 (IH, m), 3.81 (4H, m), 4.54 (IH,- dd, J= 7.0, 7.0 Hz), 5.06 (IH, d, J = 9.2 Hz), 5.14 (IH, d, J= 10.7 Hz), 5.59 (IH, dd, J= 15.2, 9.4 Hz), 5.65 (IH, dd, J= 15.2, 9.2 Hz), 5.71 (IH, dd, J= 15.1, 8.2 Hz), 6.08 (IH, d, J= 10.8 Hz), 6.30 (IH, dd, J= 15.1, 10.8 Hz), 6.51 (IH, t, J= 4.7 Hz), 8.30 (2H, d, J= 4.7 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.6, 22.2, 24.6, 25.5, 30.7, 36.2, 39.2, 40.1, 41.0, 41.6, 44.5, 44.6, 59.1, 59.2, 60.5, 70.1, 74.3, 79.1, 79.8, 83.5, 84.3, 111.3, 126.5, 127.4, 131.8, 132.7, 137.1, 141.4, 155.7, 158.7, 162.4, 170.5, 172.9; (+)-HR-ESIMS m/z 757.4343 [MH-H]⁺

GM272

¹H NMR (500 MHz, CDCl₃) <50.86 (3H, d, J= 7.3 Hz), 0.87 (3H, t, J= 7.1 Hz), 0.88 (3H, d, J= 7.1 Hz), 1.12 (3H, d, J= 6.6 Hz), 1.21 (3H, s), 1.29 (IH, m), 1.35 (IH, m), 1.36 (IH, m), 1.44 (IH, m), 1.53 (IH, m), 1.61 (IH, m), 1.64 (IH, m), 1.70 (IH, m), 1.72 (3H, s), 1.77 (IH, m), 1.85 (IH, m), 2.05 (IH, m), 2.09 (3H, s), 2.51 (IH, m), 2.53 (IH, dd, J= 14.9, 2.7 Hz), 2.60 (IH, dd, J= 14.9, 3.4 Hz), 2.70 (IH, m), 2.86 (2H, m), 2.87 (IH, m), 2.91 (IH, m), 3.11 (IH, m), 3.17 (IH, m), 3.38 (3H, s), 3.52 (IH, br s), 3.74 (IH, m), 4.19 (IH, m), 4.30 (IH, br d, J= 11.3 Hz), 4.53 (IH, dd, J= 6.6, 6.3 Hz), 5.08 (IH, d, J= 9.2 Hz), 5.15 (IH, d, J= 10.6 Hz), 5.62 (IH, dd, J= 15.2, 9.4 Hz), 5.66 (IH, dd, J= 15.2, 9.2 Hz), 5.74 (IH, dd, J= 15.1, 8.1 Hz), 6.09 (IH, d, J= 10.8 Hz), 6.31 (IH, dd, J= 15.1, 10.8 Hz), 7.16 (IH, m), 7.18 (IH, m), 7.64 (IH, t, J= 7.6 Hz), 8.56 (IH, d, J = 4.4 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.3, 12.7, 17.4, 17.6, 22.2, 24.7, 25.6, 25.9, 30.8, 31.8, 36.2, 39.3, 40.2, 41.0, 41.7, 45.0, 45.4, 50.0, 59.1, 59.4, 60.5, 70.1, 74.3, 78.6, 79.9, 83.5, 84.4, 122.8, 123.4, 126.6, 127.3, 131.9, 132.6, 137.4, 137.8, 141.4, 149.8, 155.7, 162.8, 170.6, 173.0; (+)-HR-ESIMS m/z 755.4346 [M+H]⁺

GM273

¹H NMR (500 MHz, CDCl₃) £0.86 (3H, t, J = 7.5 Hz), 0.87 (3H, d, J= 7.0 Hz), 0.89 (3H, d, J= 7.2 Hz), 1.12 (3H, d, J= 6.9 Hz), 1.21 (3H, s), 1.31 (IH, m), 1.35 (2H, m), 1.43 (IH, m), 1.53 (IH, m), 1.64 (IH, m), 1.69 (IH, m), 1.71 (3H, s), 2.09 (3H, s), 2.13 (IH, br s), 2.49 (IH, m), 2.52 (IH₅ dd, J= 15.0, 3.0 Hz), 2.61 (IH, dd, J= 15.0, 3.7 Hz), 2.68 (2H, m), 2.72 (IH, m), 2.86 (IH, m), 2.87 (IH, m), 3.17 (IH, m), 3.37 (3H, s), 3.52 (IH, d, J= 10.9 Hz), 3.73 (2H, m), 3.75 (IH, m), 4.22 (2H, m), 4.56 (IH, dd, J= 6.9, 6.9 Hz), 5.08 (IH, d, J = 9.2 Hz), 5.15 (IH, d, J= 10.6 Hz), 5.60 (IH, dd, J= 15.2, 9.4 Hz), 5.66 (IH, dd, J= 15.2, 9.2 Hz), 5.74 (IH, dd, J= 14.9, 8.2 Hz), 6.09 (IH, d, J= 11.0 Hz), 6.31 (IH, dd, J= 14.9, 11.0 Hz), 6.62 (IH, m), 7.18 (IH, dd, J= 7.0, 4.6 Hz), 7.38 (IH, d, J= 7.8 Hz), 7.68 (IH, dd, J = 7.8, 7.0 Hz), 8.57 (IH, d, J = 4.6 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.6, 22.2, 24.6, 25.5, 26.6/26.9, 30.7, 36.2, 39.2, 40.1, 41.0, 41.4/41.6, 41.6, 44.6/45.0, 59.0, 59.2, 60.5, 70.1, 74.3, 78.9, 79.8, 83.5, 84.3, 120.2, 123.1, 125.2/125.7,

126.5, 127.3, 131.8, 132.6, 135.9, 137.2, 137.9, 141.3, 149.5, 155.6/155.8, 157.5/157.7,

170.6, 172.9; (+)-HR-ESIMS m/z 753.3921 [M+H]⁺

GM274

¹H NMR (500 MHz, CDCl₃) £0.86-0.90 (9H, m), 1.12 (3H, br d, J= 6.7 Hz), 1.21 (3H, s), 1.31 (IH, m), 1.35 (IH, m), 1.38 (IH, m), 1.43 (IH, m), 1.54 (IH, m), 1.64 (IH, m), 1.68 (IH, m), 1.71 (3H, br s), 2.10 (3H, s), 2.39 (2H, m), 2.50 (IH, m), 2.53 (IH, dd, J = 15.0, 3.0 Hz), 2.62 (IH, dd, J= 15.0, 3.7 Hz), 2.74 (IH, m), 2.86 (IH, m), 2.90 (IH, dd, J= 6.7, 2.0 Hz), 3.17 (IH, m), 3.38 (3H, br s), 3.52 (IH, br d, J= 10.8 Hz), 3.64 (2H, m), 3.75 (IH, m), 4.51 (2H, m), 4.57 (IH, m), 5.08 (IH, d, J= 9.2 Hz), 5.15 (IH, br d, J= 10.4 Hz), 5.61 (IH, dd, J= 15.1, 9.3 Hz), 5.67 (IH, dd, J= 15.1, 9.2 Hz), 5.74 (IH, m), 6.09 (IH, m), 6.31 (IH, m), 6.77 (IH₅ m), 7.18 (IH, dd, J= 7.1, 4.9 Hz), 7.41 (IH, m), 7.67 (IH, br dd, J= 7.4, 7.1 Hz), 8.60 (IH, d, J = 4.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.3, 17.6, 22.2, 24.6, 25.5, 26.2/26.6, 30.7, 36.2, 39.2, 40.1, 41.0, 41.1/41.3, 41.6, 44.9/45.2, 59.1, 59.2/59.4, 60.5, 70.1, 74.2, 78.6/79.2, 79.8, 83.5, 84.3, 119.8, 123.1, 126.5, 127.2, 127.3, 131.8, 132.5, 135.2, 137.3, 137.3/137.7, 141.3, 149.7, 155.7/156.0, 156.4/156.9, 170.6, 172.9; (+)-HR-ESIMS m/z 753.4217 [M+H]⁺

GM279

¹H NMR (500 MHz, CDCl₃) £0.84 (3H, t, J= 7.5 Hz), 0.85 (3H, d, J= 7.3 Hz), 0.86 (3H, d, J= 6.7 Hz), 1.08 (3H, d, J= 6.8 Hz), 1.19 (3H, s), 1.28 (IH, m), 1.33 (IH, m), 1.34 (IH, m), 1.43 (IH, m), 1.52 (IH, m), 1.62 (IH, m), 1.68 (IH, m), 1.71 (3H, s), 2.06 (3H, s), 2.48 (IH, m), 2.51 (IH, dd, J= 15.2, 2.9 Hz), 2.58 (IH, dd, J= 15.2, 3.5 Hz), 2.64 (IH, m), 2.81 (IH, dd, J= 6.2, 2.1 Hz), 2.83 (IH, dd, J= 8.4, 2.1 Hz), 3.13 (IH, ddd, J= 6.3, 6.3, 4.2 Hz), 3.36 (3H, s), 3.54 (IH, br s), 3.73 (IH, m), 4.46 (IH, dd, J= 7.5, 6.2 Hz), 4.89 (2H, s), 5.06 (IH, d, J= 9.3 Hz), 5.12 (IH, d, J= 10.6 Hz), 5.59 (IH, dd, J= 15.2, 9.4 Hz), 5.64 (IH, dd, J= 15.2, 9.3 Hz), 5.68 (IH, dd, J= 15.1, 8.4 Hz), 6.06 (IH, d, J= 10.8 Hz), 6.30 (IH, dd, J = 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.8, 17.4, 17.7, 22.2, 24.6, 25.6, 30.7, 36.2, 39.3, 40.0, 41.1, 41.7, 59.0, 59.2, 60.4, 70.1, 74.3, 78.2, 79.9, 83.5, 84.4, 126.6, 127.5, 131.8, 132.8, 137.0, 141.4, 157.2, 170.6, 173.0; (+)-HR-ESIMS m/z 632.3375 GM284

¹H NMR (500 MHz, CDCl₃) £0.86 (3H, d, J= 7.0 Hz), 0.87 (3H, t, J= 7.0 Hz) > 0.90 (3H, d, J= 7.1 Hz), 1.13 (3H, d, J= 6.9 Hz), 1.22 (3H, s), 1.31 (IH, m), 1.36 (IH, m), 1.38 (IH, m), 1.44 (IH, m), 1.54 (IH, m), 1.65 (IH, m), 1.70 (IH, m), 1.73 (3H, s), 2.10 (3H, s), 2.15 (IH, br s), 2.50 (IH, m), 2.52 (IH, m), 2.60 (IH, dd, J= 14.9, 3.7 Hz), 2.71 (IH, m), 2.88 (2H, m), 3.01 (4H, m), 3.18 (IH, ddd, J= 6.4, 6.4, 4.0 Hz), 3.39 (3H, s), 3.56 (IH, br d, J = 10.7 Hz), 3.67 (4H, m), 3.76 (IH, m), 4.55 (IH, dd, J= 6.8, 6.8 Hz), 5.08 (IH, d, J= 9.2 Hz), 5.15 (IH, d, J = 10.6 Hz), 5.60 (IH, dd, J= 15.2, 9.5 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.73 (IH, dd, J= 15.1, 8.2 Hz), 6.09 (IH, d, J= 10.9 Hz), 6.31 (IH₅ dd, J= 15.1, 10.9 Hz), 6.78 (2H, d, J= 8.8 Hz), 6.86 (2H, br d, J = 8.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.8, 17.4, 17.5, 22.2, 24.6, 25.5, 30.7, 36.1, 39.2, 40.0, 41.0, 41.6, 44.9, 52.2, 59.1, 59.2, 60.6, 70.1, 74.4, 79.0, 79.9, 83.5, 84.4, 116.9, 120.3, 126.5, 127.4, 131.8, 132.7, 137.1, 141.4, 145.7, 152.1, 155.6, 170.7, 173.0; (+)-HR-ESIMS mfz 771.4062 [M+H]⁺

GM285

¹H NMR (500 MHz, CDCl₃) (50.87 (3H, t, J= 7.5 Hz), 0.88 (3H, d, J= 6.7 Hz), 0.88 (3H, d, J= 7.0 Hz), 1.10 (3H, d, J= 6.9 Hz), 1.21 (3H, s), 1.31 (IH, m), 1.35 (IH, m), 1.38 (IH, m), 1.43 (IH, m), 1.54 (IH, m), 1.65 (IH, m), 1.70 (IH, m), 1.74 (3H, s), 2.09 (3H, s), 2.51 (5H, m), 2.56 (IH₅ m), 2.58 (2H, m), 2.62 (IH, dd, J= 15.0, 3.7 Hz), 2.69 (IH, m), 2.85 (2H, br d, J= 7.3 Hz), 3.18 (IH, ddd, J= 6.4, 6.4, 3.9 Hz), 3.39 (3H, s), 3.54 (4H, m), 3.65 (2H, t, J = 5.4 Hz), 3.76 (IH, m), 4.52 (IH, dd, J- 7.3, 7.0 Hz), 5.08 (IH, d, J = 9.2 Hz), 5.15 (IH, d, J= 10.7 Hz), 5.61 (IH, dd, J= 15.2, 9.5 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.72 (IH, dd, J= 15.1, 8.2 Hz), 6.09 (IH, d, J= 10.9 Hz), 6.31 (IH, dd, J= 15.1, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.5, 22.2, 24.6, 25.5, 30.7, 36.2, 39.3, 40.1, 41.0, 41.6, 44.6, 53.6, 58.6, 59.1, 59.2, 60.4, 60.5, 70.1, 74.3, 78.9, 79.8, 83.5, 84.3, 126.6, 127.3, 131.8, 132.7, 137.1, 141.3, 155.5, 170.6, 172.9; (+)-HR-ESIMS m/z 723.4220 [M+H]⁺

GM286

¹H NMR (500 MHz, CDCl₃) 50.87 (3H, t, J= 7.4 Hz), 0.88 (3H, d, J= 6.7 Hz), 0.89 (3 H, d, J= 7.1 Hz), 1.10 (3H, d, J = 6.9 Hz), 1.22 (3H, s), 1.32 (IH, m), 1.35 (IH, m), 1.38 (IH, m), 1.44 (IH, m), 1.55 (IH, m), 1.63 (IH, m), 1.70 (3H, m), 1.74 (3H, s), 2.10 (3H, s), 2.28 (6H, s), 2.37-2.40 (8H, m), 2.51 (IH, m), 2.54 (IH, dd, J= 14.9, 2.9 Hz), 2.62 (IH, dd, J = 14.9, 3.7 Hz), 2.69 (IH, m), 2.85 (2H, m), 3.18 (IH, ddd, J= 6.4, 6.4, 4.0 Hz), 3.39 (3H, s), 3.51 (4H, m), 3.76 (IH, m), 4.52 (IH, dd, J= 7.0, 6.7 Hz), 5.09 (IH, d, J= 9.2 Hz), 5.16 (IH, d, J= 10.6 Hz), 5.61 (IH, dd, J= 15.2, 9.4 Hz), 5.68 (IH, dd, J= 15.2, 9.2 Hz), 5.72 (IH, dd, J= 15.1, 8.2 Hz), 6.09 (IH, d, J= 10.9 Hz), 6.31 (IH, dd, J= 15.1, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.8, 17.4, 17.6, 22.2, 24.7, 25.3, 25.6, 30.8, 36.2, 39.3, 40.1, 41.0, 41.7, 44.8, 45.9, 53.9, 57.2, 58.4, 59.1, 59.3, 60.5, 70.1, 74.3, 78.8, 79.8, 83.5, 84.4, 126.6, 127.3, 131.9, 132.6, 137.3, 141.4, 155.5, 170.6, 173.0; (+)-HR-ESIMS m/z KAA619 [M+H]⁺ GM288

¹H NMR (500 MHz, CDCl₃) £0.86 (3H, t, J= 7.7 Hz), 0.87 (3H, d, J = 7.1 Hz), 0.89 (3H, d, J= 7.0 Hz), 1.12 (3H, d, J= 6.9 Hz), 1.21 (3H, s), 1.31 (IH, m), 1.35 (IH, m), 1.37 (IH, m), 1.43 (IH, m), 1.54 (IH, m), 1.65 (IH, m), 1.70 (IH, m), 1.73 (3H, s), 2.09 (3H, s), 2.50 (IH, m), 2.52 (IH, m), 2.60 (IH, dd, J= 14.9, 3.6 Hz), 2.72 (IH, m), 2.86 (2H, br d, J= 7.1 Hz), 3.17 (IH, ddd, J= 6.3, 6.3, 4.0 Hz), 3.36 (4H, m), 3.38 (3H, s), 3.65 (4H, m), 3.75 (IH, m), 4.54 (IH, dd, J= 7.1, 6.9 Hz), 5.08 (IH, d, J= 9.3 Hz), 5.15 (IH, d, J= 10.7 Hz), 5.60 (IH, dd, J= 15.2, 9.6 Hz), 5.67 (IH, dd, J= 15.2, 9.3 Hz), 5.73 (IH, dd, J= 15.0, 8.2 Hz), 6.10 (IH, d, J= 10.9 Hz), 6.31 (IH, dd, J= 15.0, 10.9 Hz), 6.67 (2H, br d, J= 5.7 Hz), 8.30 (2H, br d, J = 5.7 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.5, 22.2, 24.5, 25.5, 30.7, 36.2, 39.3, 40.0, 40.9, 41.6, 44.0, 46.7, 59.0, 59.1, 60.6, 70.1, 74.2, 79.4, 79.8, 83.5, 84.2, 109.4, 126.7, 127.4, 131.7, 132.8, 136.9, 141.2, 150.6, 155.5, 155.7, 170.6, 172.9; (+)-HR-ESIMS m/z 756.4013 [M+H]⁺

GM289

¹H NMR (500 MHz, CDCl₃) £0.87 (3H, t, J = 7.5 Hz), 0.88 (6H, d, J= 6.7 Hz), 1.11 (3H, m), 1.22 (3H, s), 1.32 (IH, m), 1.36 (IH, m), 1.38 (IH, m), 1.44 (IH, m), 1.54 (IH, m), 1.67 (IH, m), 1.70 (IH, m), 1.74 (3H, s), 1.95 (IH, m), 1.99 (IH, m), 2.10 (3H, s), 2.51 (IH, m), 2.54 (IH, dd, J= 14.9, 3.0 Hz), 2.62 (IH, dd, J= 14.9, 3.7 Hz), 2.69 (IH, m), 2.86 (2H, br d, J= 7.0 Hz), 3.17 (IH, ddd, J= 6.4, 6.4, 4.1 Hz), 3.39 (3H, s), 3.45 (IH, m), 3.54 (4H, m), 3.75 (IH, m), 4.49 (IH, m), 4.51 (IH, m), 5.09 (IH, d, J = 9.2 Hz), 5.16 (IH, d, J = 10.7 Hz), 5.61 (IH, dd, J = 15.2, 9.4 Hz), 5.67 (IH, dd, J = 15.2, 9.2 Hz), 5.74 (IH, m), 6.09 (IH, d, J= 10.8 Hz), 6.31 (IH, dd, J= 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.5/17.6, 22.2, 24.59/24.63, 25.5, 30.7, 34.4/35.0, 36.1, 39.3, 40.0/40.2, 40.9/41.0, 41.6, 44.7/45.1, 55.1/55.7, 59.1, 59.5, 60.5/60.7, 70.1, 71.1/71.9, 74.3, 78.5/78.8, 79.8, 83.6, 84.37/84.44, 126.5, 127.2, 131.9, 132.5, 137.3/137.4, 141.4, 155.4, 170.6, 173.0; (+)-HR-ESIMS m/z 680.4004 [M+H]⁺

GM290

¹H NMR (500 MHz, CDCl₃) (50.87 (3H, t, J= 7.4 Hz), 0.88 (6H, d, J= 6.7 Hz), 1.13 (3H, d, J = 6.9 Hz), 1.22 (3H, s), 1.31 (IH, m), 1.35 (IH, m), 1.38 (IH, m), 1.44 (IH, m), 1.54 (IH, m), 1.65 (IH, m), 1.71 (IH, m), 1.74 (3H, s), 1.83 (IH, m), 1.88 (IH, m), 2.03 (2H, m), 2.10 (3H, s), 2.51 (IH, m), 2.54 (IH, dd, J= 14.9, 2.8 Hz), 2.63 (IH, dd, J= 14.9, 3.5 Hz), 2.72 (IH, m), 2.86 (2H, br d, J = 8.0 Hz), 3.18 (IH, ddd, J= 6.4, 6.4, 3.9 Hz), 3.37 (IH, m), 3.39 (3H, s), 3.59 (IH, m), 3.61 (IH, m), 3.67 (IH, m), 3.75 (IH, m), 4.01 (IH, m), 4.47 (IH, dd, J= 8.0, 6.9 Hz), 5.09 (IH, d, J= 9.2 Hz), 5.16 (IH, d, J= 10.6 Hz), 5.61 (IH, dd, J= 15.2, 9.4 Hz), 5.68 (IH, dd, J= 15.2, 9.2 Hz), 5.73 (IH, dd, J= 15.1, 8.0 Hz), 6.10 (IH, d, J= 10.7 Hz), 6.32 (IH, dd, J= 15.1, 10.7 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.36, 17.41, 22.2, 24.6, 24.9, 25.5, 29.6, 30.7, 36.2, 39.3, 40.1, 41.0, 41.6, 48.2, 59.1, 59.3, 60.7, 61.7, 68.0, 70.1, 74.3, 79.6, 79.8, 83.5, 84.3, 126.6, 127.3, 131.8, 132.7, 137.0, 141.3, 157.6, 170.6, 173.0; (+)-HR-ESIMS m/z 716.3656 [M+Naf GM291

¹H NMR (500 MHz, CDCl₃) £0.84 (3 H, t, J = 7.6 Hz), 0.85 (3H, m), 0.86 (3H, d, J= 6.9 Hz), 1.09 (3H, d, J- 6.9 Hz), 1.19 (3H, s), 1.28 (IH, m), 1.33 (IH, m), 1.36 (IH, m), 1.42 (IH, m), 1.53 (IH, m), 1.63 (IH, m), 1.69 (IH, m), 1.71 (3H, s), 2.07 (3H, s), 2.50 (IH, m), 2.52 (IH, dd, J= 14.8, 2.7 Hz), 2.60 (IH, dd, J= 14.8, 3.6 Hz), 2.67 (IH, m), 2.83 (6H, m), 3.16 (IH, ddd, J= 6.3, 6.3, 4.0 Hz)₅ 3.37 (3H, s), 3.46 (4H, m), 3.73 (IH, m), 4.50 (IH, dd, J= 6.8, 6.6 Hz), 5.07 (IH, d, J= 9.3 Hz), 5.13 (IH, d, J= 10.6 Hz), 5.59 (IH, dd, J= 15.2, 9.5 Hz), 5.65 (IH, dd, J= 15.2, 9.3 Hz), 5.70 (IH, dd, J= 15.1, 8.2 Hz), 6.07 (IH, d, J = 10.8 Hz), 6.29 (IH, dd, J= 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) 510.9, 11.2, 12.7, 17.4, 17.5, 22.2, 24.6, 25.5, 30.7, 36.2, 39.3, 40.1, 41.0, 41.6, 45.7, 46.6, 59.1, 59.2, 60.4, 70.1, 74.3, 78.8, 79.8, 83.5, 84.3, 126.6, 127.3, 131.8, 132.6, 137.2, 141.3, 155.6, 170.6, 172.9; (+VHR-ESIMS m/z 679.3955 [M+H]⁺

GM292

¹H NMR (500 MHz, CDCl₃) <50.87 (3H, m), 0.88 (6H, m), 1.09 (3H, m), 1.21 (3H, s), 1.33 (IH, m), 1.36 (IH, m), 1.39 (IH, m), 1.44 (IH, m), 1.54 (IH, m), 1.65 (IH, m), 1.70 (IH, m), 1.73 (3H, br s), 2.09 (3H, s), 2.13 (IH, br s), 2.50 (IH, m), 2.53 (IH, dd, J= 15.0, 3.0 Hz), 2.61 (IH, m), 2.68 (IH, m), 2.84 (2H, m), 2.88 (3H, br s), 3.08 (2H, m), 3.18 (IH, ddd, J = 6.3, 6.3, 4.1 Hz), 3.39 (3H, m), 3.53 (IH, br d, J = 10.7 Hz), 3.68 (2H, m), 3.74 (IH, m), 4.49 (IH, m), 5.08 (IH, d, J= 9.2 Hz), 5.15 (IH, d, J= 10.6 Hz), 5.60 (IH, dd, J = 14.9, 9.7 Hz), 5.67 (IH, dd, J= 14.9, 9.2 Hz), 5.72 (IH, m), 6.09 (IH, d, J= 10.8 Hz), 6.30 (IH, m), 7.21 (IH, m), 7.29 (IH, m), 7.66 (IH, m), 8.55 (IH, m); ¹³C NMR (125 MHz, 4214

CDCl₃) δ 10.9, 11.3, 12.7, 17.3, 17.5/17.6, 22.2, 24.6, 25.5, 30.7, 35.5/36.0, 36.1, 36.3/37.4, 39.2, 40.1, 41.0/41.1, 41.6, 49.9/50.3, 59.1, 59.2/59.3, 60.5, 70.1, 74.3, 78.9/79.0, 79.8, 83.5, 84.3, 122.9/123.1, 125.1/125.6, 126.6, 127.3, 131.8, 132.6, 137.3, 138.7/139.6, 141.3, 148.1/149.2, 156.5/156.6, 159.1/159.4, 170.6, 172.9; (+)-HR-E3IMS «/£ 729.4214 [MH-H]⁺

GM293

¹B. NMR (500 MHz, CDCl₃) £0.87 (3H, t, J= 8.1 Hz), 0.88 (6H, d, J= 7.0 Hz), 1.12 (3H, d, J = 6.8 Hz), 1.21 (3H, s), 1.31 (IH, m), 1.38 (2H, m), 1.43 (IH, m), 1.54 (IH, m), 1.66 (IH, m), 1.71 (IH, m), 1.74 (3H, s), 2.10 (3H, s), 2.51 (IH, m), 2.54 (IH, dd, J = 14.9, 2.7 Hz), 2.62 (IH, dd, J= 14.9, 3.5 Hz), 2.71 (IH, m), 2.86 (2H, m), 3.00 (3H, s), 3.17 (IH, ddd, J= 6.3, 6.3, 4.1 Hz), 3.39 (3H, m), 3.46 (2H, m), 3.77 (3H, m), 4.52 (IH, m), 5.08 (IH, d, J= 9.2 Hz), 5.16 (IH, d, J= 10.7 Hz), 5.61 (IH, dd, J= 15.2, 9.4 Hz), 5.67 (IH, dd, J = 15.2, 9.2 Hz), 5.73 (IH, m), 6.09 (IH, d, J= 10.9 Hz), 6.32 (IH, dd, J= 15.0, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) 5 10.9, 11.1, 12.7, 17.3, 17.5/17.6, 22.2, 24.5, 25.5, 30.7, 36.1, 36.3/36.7, 39.3, 39.9, 40.9/41.1, 41.6, 51.9/52.9, 59.1, 59.2, 60.4/60.7, 61.6/62.1, 70.1, 74.3, 78.8/79.0, 79.8, 83.5, 84.4, 126.5, 127.3, 131.8, 132.7, 137.1, 141.3, 156.8/157.9, 170.6, 173.0; (+)-HR-ESIMS m/z 690.3645 [M+Naf

GM294

¹H NMR (500 MHz, CDCl₃) 50.86 (3H, t, J= 7.5 Hz), 0.88 (6H, d, J= 6.8 Hz), 1.10 (3H, d, J = 6.9 Hz), 1.21 (3H, s), 1.28 (IH, m), 1.32 (IH, m), 1.36 (IH, m), 1.43 (IH, m), 1.54 (IH, m), 1.64 (IH, m), 1.70 (IH, m), 1.73 (3H, s), 1.80 (2H, m), 2.10 (3H, s), 2.30 (3H, s),

2.34 (3H, s), 2.41 (2H, m), 2.50 (IH, m), 2.53 (IH, dd, J= 15.0, 2.8 Hz), 2.62 (IH, dd, J = T/IB2006/004214

15.0, 3.6 Hz), 2.68 (IH, m), 2.83 (2H, m), 2.93 (3H, s), 3.16 (IH, m), 3.33 (2H, m), 3.38 (3H, s), 3.75 (IH, m), 4.48 (IH, dd, J= 7.0, 7.0 Hz), 5.08 (IH, d, J = 9.2 Hz), 5.15 (IH, d, J = 10.7 Hz), 5.61 (IH, dd, J = 15.2, 9.4 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.72 (IH, dd, J = 15.0, 8.2 Hz), 6.09 (IH, d, J= 10.7 Hz), 6.31 (IH, dd, J= 15.0, 10.7 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.3, 17.5/17.7, 22.2, 24.5, 25.5, 26.1/26.7, 30.7, 34.9/35.8, 36.2, 39.2, 40.1, 41.0/41.1, 41.6, 45.88, 45.93, 47.7/48.1, 57.5/57.6, 59.0, 59.3, 60.5/60.6, 70.0, 74.2, 78.8/79.0, 79.8, 83.5, 84.3, 126.5, 127.2, 131.8, 132.6, 137.2, 141.3, 156.6/156.7, 170.6, 172.9; (+)-HR-ESIMS m/z 709.4425 [M+H]⁺

GM296

¹H NMR (500 MHz, CDCl₃) 50.84 (3 H, t, J= 7.6 Hz), 0.85 (3H, d, J= 6.7 Hz), 0.86 (3 H, d, J= 7.0 Hz), 1.09 (3H, d, J= 6.6 Hz), 1.20 (3H, s), 1.32 (IH, m), 1.34 (IH, m), 1.36 (IH, m), 1.44 (IH, m), 1.53 (IH, m), 1.63 (IH, m), 1.69 (IH, m), 1.72 (3H, s), 2.07 (3H, s), 2.25 (3H , s), 2.28 (3H , s), 2.48 (2H, m), 2.50 (IH, m), 2.52 (IH, dd, J= 14.8, 2.6 Hz), 2.60 (IH, dd, J= 14.8, 3.2 Hz), 2.67 (IH, ni), 2.81 (IH, m), 2.83 (IH, m), 2.92 (3H, s), 3.15 (IH, ddd, J= 6.3, 6.3, 4.0 Hz), 3.37 (3H, s), 3.40 (2H, m), 3.73 (IH, m), 4.48 (IH, m), 5.06 (IH, d, J = 9.3 Hz), 5.13 (IH, d, J= 10.7 Hz), 5.59 (IH, dd, J= 15.2, 9.5 Hz), 5.65 (IH, dd, J= 15.2, 9.3 Hz), 5.70 (IH, dd, J= 15.1, 8.3 Hz), 6.06 (IH, d, J= 10.7 Hz), 6.28 (IH, dd, J= 15.1, 10.7 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.7, 22.2, 24.6, 25.5, 30.7, 35.4, 36.2, 39.3, 40.1, 41.0/41.2, 41.6, 46.1, 46.5, 47.7, 57.1 / 58.1, 59.1, 59.3, 60.5, 70.1, 74.3, 78.9/79.0, 79.8, 83.5, 84.3, 126.5, 127.3, 131.8, 132.6, 137.27/137.35, 141.3, 156.5/156.7, 170.6, 172.9; (+)-HR-ESIMS m/z 695.4288 [M+H]⁺ 14

GM297

¹H NMR (500 MHz, CDCl₃) δ 0.85-0.88 (9H, m), 1.13 (3H, d, J = 6.9 Hz), 1.21 (3H, s), 1.31 (IH, m), 1.36 (IH, m), 1.37 (IH, m), 1.43 (IH, m), 1.54 (IH, m), 1.65 (IH, m), 1.71 (IH, m), 1.74 (3H, s), 1.92 (2H, m), 2.10 (3H, s), 2.51 (IH, m), 2.54 (IH, dd, J= 14.9, 2.8 Hz), 2.62 (IH, dd, J= 14.9, 3.6 Hz), 2.74 (IH, m), 2.88 (IH, m), 2.90 (IH, m), 3.17 (IH, ddd, J= 6.5, 6.5, 3.9 Hz), 3.39 (3H, s), 3.47 (2H, m), 3.68 (2H, m), 3.76 (IH, m), 4.57 (IH, dd, J= 7.5, 7.4 Hz), 5.08 (IH, d, J= 9.3 Hz), 5.15 (IH, d, J= 10.7 Hz), 5.61 (IH, dd, J = 15.2, 9.5 Hz), 5.66 (IH, m), 5.68 (IH, m), 6.08 (IH, d, J= 10.9 Hz), 6.33 (IH, dd, J= 15.1, 10.9 Hz), 8.20 (IH, s); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.1, 12.7, 17.3, 17.6, 21.1, 22.2, 24.4, 25.5, 30.7, 36.2, 39.3, 39.9, 40.8, 41.6, 42.1, 44.2, 58.9, 59.0, 61.0, 70.1, 74.3, 79.8, 81.7, 83.4, 84.2, 126.6, 127.9, 131.5, 133.4, 136.0, 141.3, 144.9, 154.2, 170.6, 172.9; (+)-HR-ESIMS m/z 677.3750 [M+H]⁺

GM301

¹H NMR (500 MHz, CDCl₃) £0.87 (3H, t, J= 7.4 Hz), 0.88 (3H, d, J= 6.7 Hz), 0.89 (3H, d, J= 7.0 Hz), 1.11 (3H, d, J= 6.9 Hz), 1.21 (3H, s), 1.31 (IH, m), 1.35 (IH, m), 1.38 (IH, m), 1.43 (IH, m), 1.50 (2H, m), 1.54 (IH, m), 1.65 (IH, m), 1.70 (IH, m), 1.73 (3H, s), 1.87 (2H, m), 2.10 (3H, s), 2.51 (IH, m), 2.53 (IH, dd, J= 14.9, 2.9 Hz), 2.62 (IH, dd, J= 14.9, 3.7 Hz), 2.69 (IH, m), 2.85 (2H, m), 3.13 (2H, m), 3.18 (IH, m), 3.39 (3H, s), 3.75 (IH, m), 3.88 (IH, m), 3.91 (2H, m), 4.52 (IH, dd, J = 7.0, 6.8 Hz), 5.08 (IH, d, J = 9.2 Hz), 5.15 (IH, d, J= 10.7 Hz), 5.61 (IH, dd, J= 15.2, 9.3 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.73 (IH, dd, J= 15.1, 8.2 Hz), 6.09 (IH, d, J= 10.8 Hz), 6.31 (IH, dd, J= 15.1, 10.8 Hz);

¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.1, 12.7, 17.3, 17.4, 22.2, 24.6, 25.5, 30.6, 34.9, 36.1, 39.2, 40.0, 40.9, 41.6, 42.3, 59.0, 59.2, 60.3, 68.1, 70.0, 74.2, 78.6, 79.8, 83.5, 84.3, 126.5, 127.2, 131.8, 132.5, 137.2, 141.3, 155.5, 170.6, 172.9; (+)-HR-ESIMS m/z 716.3818 [M+Na]⁺

GM302

¹H NMR (500 MHz, CDCl₃) δ 0.85-0.89 (9H, m), 1.11 (3H, d, J = 6.8 Hz), 1.21 (3H, s), 1.30 (IH, m), 1.35 (IH, m), 1.37 (IH, m), 1.43 (IH, m), 1.54 (IH, m), 1.64 (IH, m), 1.69 (IH, m), 1.73 (3H, s), 1.86 (2H, m), 2.09 (3H, s), 2.50 (IH, m), 2.53 (IH, dd, J= 14.9, 2.9 Hz), 2.62 (IH, dd, J= 14.9, 3.5 Hz), 2.70 (IH, m), 2.86 (2H, m), 2.94 (2H, m), 2.98 (2H, m), 3.18 (IH, m), 3.39 (3H, s), 3.54 (4H, m), 3.75 (IH, m), 4.54 (IH, m), 5.08 (IH, d, J = 9.2 Hz), 5.14 (IH, d, J= 10.7 Hz), 5.60 (IH, dd, J= 15.2, 9.5 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.73 (IH, m), 6.09 (IH, d, J= 10.9 Hz), 6.31 (IH, dd, J= 15.0, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) 5 10.9, 11.1, 12.6/12.7, 17.3, 17.46/17.52, 22.2, 24.5, 25.4, 30.0/30.1, 30.6, 36.2, 39.3, 40.0, 41.0, 41.5, 46.5/46.9, 48.2/48.5, 49.3/49.6, 49.8/49.9, 59.0, 59.2/59.3, 60.4/60.5, 70.0, 74.2, 78.66/78.74, 79.8, 83.5, 84.2, 126.5, 127.2, 131.72/131.74, 132.59/132.64, 137.17/137.20, 141.2, 156.2/156.3, 170.6, 172.9; (+)-HR-ESIMS m/z 693.4154 [MH-H]⁺

Scheme 6 Procedure for the Syntheses of N831 (A) Rn Esters

R₁₇ ester R' = H, R" = COR ave yield 60% R₃, R₁₇ diester R" = R" = COR ave yield 10%

To a solution of N831 (A) (20 mg, 0.04 mmol), DMAP (1 mg, 8.2 μmol) and a base (triethylamine or pryridine, 0.04 mmol) in anhydrous CH₂Cl₂ (10 mL) was added the acylating reagent (0.04 mmol). The reaction mixture was stirred at room temperature for 30 min. Additional acylating reagent (0.04 mmol) and base (0.04 mmol) were added twice over a period of 1 hour. The mixture was concentrated and purified by preparative TLC (ethyl acetate-hexane 1:1) to give the C3, C17-diester and C17-monoester.

For GM203, the Fmoc protecting group was removed by stirring the acylated product in 20% piperidine in chloroform at room temperature for 1 hour. The solvents were removed under reduced pressure and the mixture purified by preparative TLC (chloroform- methanol 20:1). The deprotection yield for GM 203 was 41%.

GMl 15

¹H NMR (500 MHz, CDCl₃) <50.87 (3H, t, J= 7.4 Hz), 0.87 (3H, d, J= 7.1 Hz), 0.89 (3H, d, J= 6.8 Hz), 1.07 (3H, d, J= 6.9 Hz), 1.21 (3H, s), 1.33 (IH, m), 1.44 (IH, m), 1.45 (IH, m), 1.56 (2H, m), 1.63 (IH, m), 1.72 (3H, s), 1.75 (IH, m), 2.09 (3H, s), 2.10 (3H, s), 2.11 (3H, s), 2.53 (IH, m), 2.53 (IH, dd, J= 14.7, 3.4 Hz), 2.64 (IH, m), 2.68 (IH, dd, J= 14.7, 4.0 Hz), 2.83 (2H, br d, J= 7.6 Hz), 3.17 (IH, ddd, J= 6.4, 6.4, 3.9 Hz), 3.39 (3H, s), 4.57 (IH, dd, J= 7.6, 6.7 Hz), 4.87 (IH, m), 5.00 (IH, d, J= 10.7 Hz), 5.08 (IH, d, J= 9.3 Hz), 5.63 (IH, dd, J= 15.2, 9.7 Hz), 5.65 (IH, dd, J= 15.0, 7.8 Hz), 5.70 (IH, dd, J= 15.2, 10.7 Hz), 6.10 (IH, d, J = 10.9 Hz), 6.30 (IH, dd, J = 15.0, 10.9 Hz); ¹³C NMR (125 MHz, CDCl₃) (59.9, 10.2, 11.9, 16.3, 16.8, 21.0, 21.2, 23.6, 24.7, 26.0, 29.7, 34.6, 36.4, 39.1, 39.8, 40.5, 58.1, 58.2, 59.9, 70.8, 73.5, 78.8, 82.9, 83.4, 125.4, 126.7, 130.8, 132.1, 135.6, 140.7, 168.6, 169.6, 170.3, 170.5; (+)-HR-ESIMS m/z 673.3652 [M+Naf

GM116

¹H NMR (500 MHz, CDCl₃) 50.86 (3H, t, J= 7.4 Hz), 0.87 (3H, d, J= 6.9 Hz), 0.88 (3H, d, J= 6.7 Hz), 1.08 (3H, d, J= 6.9 Hz), 1.21 (3H, s), 1.28 (IH, m), 1.34 (IH, m), 1.36 (IH, m), 1.43 (IH, m), 1.54 (IH, m), 1.64 (IH, m), 1.67 (IH, m), 1.74 (3H, s), 2.09 (3H, s), 2.11 (3H, s), 2.50 (IH, m), 2.53 (IH, dd, J= 14.9, 2.9 Hz), 2.62 (IH, dd, J= 14.9, 3.6 Hz), 2.65 (IH, m), 2.83 (2H, br d, J= 7.3 Hz), 3.16 (IH, ddd, J= 6.4, 6.4, 3.9 Hz), 3.38 (3H, s), 3.52 (IH, br d, J= 10.2 Hz), 3.75 (IH, m), 4.57 (IH, dd, J= 7.3, 7.1 Hz), 5.08 (IH, d, J= 9.2 Hz), 5.15 (IH, d, J= 10.6 Hz), 5.60 (IH, dd, J= 15.2, 9.4 Hz), 5.67 (IH, dd, J= 15.2, 9.2 Hz), 5.68 (IH, dd, J= 15.0, 8.4 Hz), 6.08 (IH, d, J= 10.7 Hz), 6.31 (IH, dd, J= 15.0, 10.7 Hz); ¹³C NMR (125 MHz, CDCl₃) 59.9, 10.2, 11.8, 16.4, 16.7, 20.9, 21.2, 23.6, 24.6, 29.8, 35.2, 38.3, 39.1, 39.8, 40.7, 58.1, 58.2, 59.8, 69.1, 73.4, 78.8, 82.5, 83.4, 125.6, 126.6, 130.8, 131.9, 135.9, 140.4, 169.6, 170.3, 172.0; (+)-HR-ESIMS m/z 631.3420 [M+Naf

GM140

¹H NMR (500 MHz, CDCl₃) 50.87 (3H, t, J= 7.5 Hz), 0.89 (3H, d, J= 7.3 Hz), 0.91 (3H, m), 1.17 (3H, d, J= 6.8 Hz), 1.22 (3H, s), 1.30 (IH, m), 1.39 (IH, m), 1.40 (IH, m), 1.46 (IH, m), 1.55 (IH, m), 1.64 (IH, m), 1.71 (IH, m), 1.72 (3H, s), 2.10 (3H, s), 2.51 (IH, m), 2.55 (IH, dd, J= 15.0, 2.4 Hz), 2.62 (IH, dd, J= 15.0, 3.5 Hz), 2.82 (IH, m), 2.90 (IH, br d, J= 8.2 Hz), 2.99 (IH, br d, J= 6.7 Hz), 3.17 (IH, m), 3.38 (3H, s), 3.75 (IH, m), 4.85 (IH, dd, J= 7.2, 6.7 Hz), 5.09 (IH, d, J= 9.1 Hz), 5.16 (IH, d, J= 10.7 Hz), 5.62 (IH, dd, J = 15.2, 9.2 Hz), 5.65 (IH, dd, J= 15.2, 9.1 Hz), 5.78 (IH, dd, J= 15.1, 8.2 Hz), 6.11 (IH, d, J= 10.8 Hz), 6.35 (IH, dd, J= 15.1, 10.8 Hz), 7.45 (2H, t, J= 7.7 Hz), 7.57 (IH, t, J = 7.7 Hz), 8.07 (2H, d, J = 7.7 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.3, 12.7, 17.4, 17.7, 22.3, 24.7, 25.6, 30.8, 36.2, 39.3, 40.2, 41.0, 41.7, 59.1, 59.2, 60.7, 70.1, 74.4, 78.5, 79.9, 83.5, 84.4, 126.6, 127.7, 129.3, 130.8, 131.0, 131.8, 132.9, 134.1, 136.9, 141.4, 166.8, 170.6, 173.0; (+)-HR-ESIMS m/z 6113116 [M+H]⁺ GM203

¹H NMR (500 MHz, CDCl₃) 50.86 (3H, m), 0.86 (3H, m), 0.87 (3H, m), 1.09 (3H, d, J =

6.8 Hz), 1.21 (3H, s), 1.32 (IH, m), 1.35 (IH, m), 1.38 (IH, m), 1.44 (IH, m), 1.55 (IH, m), 1.64 (IH, m), 1.71 (IH, m), 1.74 (3H, s), 2.09 (3H, s), 2.51 (IH, m), 2.54 (IH, dd, J- 14.9,

2.9 Hz), 2.61 (IH, dd, J= 14.9, 3.6 Hz), 2.67 (IH, m), 2.83 (IH, dd, J= 5.4, 2.1 Hz), 2.85 (IH, dd, J= 8.1, 2.1 Hz), 3.16 (IH, ddd, J= 6.4, 6.4, 4.0 Hz), 3.38 (3H, s), 3.46 (IH, d, J = 18.2 Hz), 3.52 (IH, d, J= 18.2 Hz), 3.75 (IH, m), 4.61 (IH, dd, J= 7.4, 5.4 Hz), 5.09 (IH, d, J= 9.3 Hz), 5.15 (IH, d, J= 10.6 Hz), 5.60 (IH, dd, J= 15.2, 9.4 Hz), 5.66 (IH, dd, J = 15.1, 8.4 Hz), 5.68 (IH, dd, J= 15.2, 9.3 Hz), 6.09 (IH, d, J= 10.8 Hz), 6.31 (IH, dd, J = 15.1, 10.8 Hz)

GM263

¹H NMR (500 MHz, CDCl₃) 50.86 (3H, d, J = 7.3 Hz), 0.87 (3H, m), 0.88 (5H, ni), 1.02 (2H, m), 1.08 (3H, d, J= 6.8 Hz), 1.21 (3H, s), 1.28 (IH, m), 1.33 (IH, m), 1.38 (IH, m), 1.43 (IH, m), 1.54 (IH, br t, J = 13.0 Hz), 1.64 (IH, m), 1.65 (IH, m), 1.69 (IH, m), 1.73 (3H, s), 2.09 (3H, s), 2.50 (IH, m), 2.53 (IH, dd, J= 14.9, 2.6 Hz), 2.62 (IH, dd, J =14.9, 3.4 Hz), 2.66 (IH, m), 2.82 (IH, dd, J= 8.3, 1.9 Hz), 2.84 (IH, dd, J = 6.8, 1.9 Hz), 3.17 (IH, ddd, J= 6.3, 6.3, 4.1 Hz), 3.38 (3H, s), 3.75 (IH, m), 4.58 (IH, dd, J= 7.2, 6.8 Hz), 5.08 (IH, d, J= 9.2 Hz), 5.15 (IH, d, J= 10.7 Hz), 5.60, (IH, dd, J= 15.1, 9.4 Hz), 5.67 (IH, dd, J= 15.1, 9.2 Hz), 5.69, (IH, dd, J= 15.1, 8.8 Hz), 6.08 (IH, d, J= 10.8 Hz). 6.31 (IH, dd, J= 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) 59.5, 9.6, 10.9, 11.2, 12.8, 13.9,

17.4, 17.7, 22.2, 24.6, 25.6, 30.8, 36.2, 39.3, 40.1, 40.8, 41.7, 59.1, 59.1, 60.6, 70.1, 74.4, 77.6, 79.9, 83.5, 84.4, 126.6, 127.5, 131.8, 132.8, 137.0, 141.4, 170.6, 173.0, 175.1; (+)- HR-ESIMS m/z 657.1785 [M+Na]⁺

GM268

¹H NMR (500 MHz, CDCl₃) 50.86 (3H, t, J = 7.4 Hz), 0.89 (3H, d, 3H, J = 6.5 Hz), 0.90 (3H, d, J = 7.1 Hz), 1.16 (3H, d, J= 6.9 Hz), 1.22 (3H, s), 1.30 (IH, m), 1.39 (2H, m), 1.44 (IH, m), 1.54 (IH, m), 1.64 (IH, m), 1.71 (IH, m), 1.72 (3H, s), 2.09 (3H, s), 2.52 (IH, m), 2.53 (IH, dd, J= 15.0, 3.0 Hz), 2.62 (IH, dd, J= 15.0, 3.7 Hz), 2.82 (IH, m), 2.91 (IH, dd, J = 8.2, 2.0 Hz), 2.98 (IH, dd, J= 7.2, 2.0 Hz), 3.16 (IH, ddd, J= 6.5, 6.4, 3.9 Hz), 3.38 (3H, s), 3.75 (IH, m), 4.83 (IH, dd , J= 7.3, 7.2 Hz), 5.08 (IH, d, J= 9.3 Hz), 5.15 (IH, d, J= 10.7 Hz), 5.60, (IH, dd, J= 15.2, 9.5 Hz), 5.68 (IH, dd, J= 15.2, 9.3 Hz), 5.73, (IH, dd, J= 15.1, 8.3 Hz), 6.10 (IH, d, J= 10.8 Hz). 6.35 (IH, dd, J= 15.1, 10.8 Hz), 7.89 (2H, br d, J = 3.5 Hz), 8.80 (2H, br s); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.7, 17.4, 17.7, 22.2, 24.5, 25.6, 30.8, 36.2, 39.3, 40.0, 40.9, 41.7, 58.9, 59.0, 61.0, 70.1, 74.3, 79.8, 79.9, 83.4, 84.3, 124.0, 126.7, 128.0, 131.5, 133.4, 136.2, 138.3, 141.3, 151.4, 165.3, 170.6, 173.0; (+)-HR-ESIMS m/z 672.4769 [M+H]⁺; (+)-HR-ESIMS m/z 694.3926 [M+Na]⁺

Scheme 7 Procedure for the Synthesis of N1407 Ri 7 Carbamate

GW1222

29%

To a solution of compound 4 (111 mg, 0.15 rnniol, obtained by protection of GMl 16 with ethyl vinyl ether) in methanol (6 mL) was added anhydrous K₂CO₃ (60 mg, 0.43 mmol). The reaction mixture was stirred at room temperature for 4.5 hours and then concentrated under reduced pressure. The residue was partitioned between ethyl acetate (6 mL) and water (4 mL), and extracted with ethyl acetate (3 x 6 mL). The organic extracts were combined, dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (ethyl acetate-hexane 1:1) to give compound 5 as a colourless oil (56 mg, 57%).

To a solution of compound 5 (50 mg, 0.08 mmol), DMAP (4.5 mg, 0.04 mmol) and triethylamine (125 μL, 0.90 mmol) in anhydrous CH₂Cl₂ (5 mL) was added 4- nitrophenylchloroformate (90 mg, 0.45 mmol). The reaction mixture was stirred at room temperature for 2 hours. Additional 4-nitrophenylchloroformate (30 mg, 0.15 mmol) was added. The mixture was stirred at room temperature for another 16 hours, and then washed with saturated aqueous NaHCO₃ (3 x 6 mL). The organic layer was dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure to give crude carbonate. To a solution of crude carbonate (0.08 mmol) in anhydrous THF (5 mL) was added 1- methylpiperazine (42 μL, 0.38 mmol). The reaction mixture was stirred at room temperature for 23.5 hours and then concentrated under reduced pressure. The crude product was purified by preparative TLC (ethyl acetate-hexane 2: 1, then cliloroform-methanol 50: 1 , 40: 1 then 30:1) to give compound 6 as a colourless oil (8.5 mg, 14%).

To a solution of compound 6 (8.5 mg, 0.01 mmol) in methanol (2 niL) was added pyridinium/)-toluenesulfonate (3 mg, 0.01 mmol). The reaction mixture was stirred at room temperature for 1.5 hours and then concentrated under reduced pressure. Additional methanol (2 mL) was added. The reaction mixture was stirred at room temperature for 1 hour and then concentrated under reduced pressure. The addition and removal of methanol (2 mL) was repeated another 2 times over 18 hours. The crude product was purified by preparative TLC (chloroform-methanol 30:1) to give GM222 as a colourless oil (2 mg, 29%).

GM222

¹H NMR (500 MHz, CDCl₃) δ 0.88 (3H, t, J= 7.5 Hz), 0.89 (3H, d, J= 6.7 Hz), 0.90 (3H, m), 1.12 (3H, d, J= 6.9 Hz), 1.27 (IH, m), 1.32 (3H, s), 1.36 (IH, m), 1.40 (IH, m), 1.44 (IH, m), 1.49 (IH, m), 1.64 (IH, m), 1.69 (IH, m), 1.75 (3H, s), 2.36 (3H, s), 2.45 (4H, m), 2.53 (IH, m), 2.56 (IH, dd, J= 14.9, 3.0 Hz), 2.61 (IH, dd, J= 14.9, 3.6 Hz), 2.69 (IH, m), 2.85 (2H, br d, J= 7.1 Hz), 3.18 (IH, ddd, J= 6.5, 6.5, 3.8 Hz), 3.40 (3H, s), 3.57 (4H, m), 3.75 (IH, m), 3.81 (IH, d, J= 9.7 Hz), 4.53 (IH, dd, J= 7.1, 7.1 Hz), 5.15 (IH, d, J= 10.7 Hz), 5.41 (IH, dd, J= 15.1, 9.9 Hz), 5.72 (IH, dd, J= 15.0, 8.8 Hz), 5.73 (IH, dd, J= 15.1, 9.7 Hz), 6.09 (IH, d, J= 10.8 Hz), 6.31 (IH, dd, J = 15.0, 10.8 Hz); (+)-HR-ESIMS m/z 651.4203 [M+H]⁺ Scheme 8 Procedure for the Synthesis of N1407 RΛ Carbamate

GM167

34%

To a solution of N1407 (11 mg, 0.02 mmol) and NaH (2.6 mg, 0.07 mmol, 60% in oil) in anhydrous THF (2 mL) was added dimethylcarbamyl chloride (6 μL, 0.07 mmol). The reaction mixture was stirred at room temperature for 14 hours. Additional anhydrous THF (2 mL), NaH (17 mg, 0.43 mmol, 60% in oil) and dimethylcarbamyl chloride (40 μL, 0.43 mmol) were introduced. The mixture was stirred at room temperature for another 3 days and then concentrated under reduced pressure. The residue was partitioned between ethyl acetate (4 mL), saturated aqueous NH₄Cl (1 mL) and water (3 mL), and extracted with ethyl acetate (3 x 4 mL). The organic extracts were combined, dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by preparative TLC (chloroform-methanol 20:1, developed thrice) to give GMl 67 (4.2 mg, 34%).

GM167

¹H NMR (500 MHz, CDCl₃) 0.88 (3H, d, J = 7.6 Hz), 0.89 (3H, m), 50.90 (3H, t, J = 7.2 Hz), 1.14 (3H, d, J = 6.8 Hz), 1.31 (3H, s), 1.41 (IH, m), 1.46 (2H, m), 1.47 (IH, m), 1.62 (IH, m), 1.64 (IH, m), 1.65 (IH, m), 1.73 (3H, s), 2.48 (IH, m), 2.54 (IH, m), 2.55 (IH, dd, J= 14.7, 3.2 Hz), 2.65 (IH, dd, J= 14.7, 3.8 Hz), 2.80 (IH, dd, J= 5.3, 2.1 Hz), 2.89 (IH, m), 2.91 (3H, s), 2.96 (3H, s), 3.18 (IH, ddd, J= 6.4, 6.4, 3.9 Hz), 3.29 (IH, dd, J= 5.8, 5.3 Hz), 3.41 (3H, s), 3.80 (IH, d, J = 9.7 Hz), 4.79 (IH, m), 5.01 (IH, d, J = 10.7 Hz), 5.46 (IH, dd, J= 15.1, 9.9 Hz), 5.65 (IH, dd, J= 15.0, 8.7 Hz), 5.74 (IH, dd, J= 15.1, 9.7 Hz), 6.09 (IH, d, J = 10.7 Hz), 6.30 (IH, dd, J = 15.0, 10.7 Hz); (+)-HR-ESIMS nι/z 618.3628 Scheme 9 Procedure for the Synthesis of N831 (A) Rig Carbamate

GM 220

14%

GM120 (130 mg, 0.20 mmol) was dissolved in dry CH₂Cl₂ (4 mL) prior to the addition of triethylamine (200 μL, 1.98 mmol). The reaction mixture was stirred at 50 ⁰C for 48 hours. The solvent was then removed under reduced pressure and the crude product purified by preparative TLC (cliloroform-methanol 99:1, then 49:1) to give GM179 (79.8 mg, 70%).

To a solution of GM179 (55 mg, 0.10 mmol), DMAP (6 mg, 0.05 mmol) and triethylamine (41 μL, 0.29 mmol) in anhydrous THF (5 mL) was added chlorotriethylsilane (51 μiL, 0.30 mmol) portion-wise over 5 hours at room temperature. The mixture was then quenched with water (1 mL). The bulk of THF was removed under reduced pressure. The residue was extracted with ethyl acetate (3 x 5 mL). The organic extracts were combined, dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (gradient elution with ethyl acetate-hexane 1:2 to ethyl acetate only) to give compound 7 as a colourless oil (40 mg, 61%).

To a solution of compound 7 (40 mg, 0.06 mmol), DMAP (3.6 mg, 0.03 mmol) and triethylamine (25 μL, 0.18 mmol) in anhydrous CH₂Cl₂ (5 mL) was added 4-nitrophenyl chlorofoπnate (36 mg, 0.18 mmol). The reaction mixture was stirred at room temperature for 17 hours. Additional 4-nitrophenyl chloroformate (24 mg, 0.12 mmol) and triethylamine (17 μL, 0.12 mmol) were added. The mixture was stirred at room temperature for another 2 hours and concentrated under reduced pressure. The residue was taken into ethyl acetate (10 mL) and washed with saturated aqueous NaHCO₃ (3 x 8 mL). The organic layer was dried with anhydrous MgSO₄, filtered, and concentrated under reduced pressure to give the crude carbonate. To the crude carbonate (0.06 mmol) in anhydrous THF (4 mL) was added 1- methylpiperazine (20 μL, 0.18 mmol). The reaction mixture was stirred at room temperature for 16 hours and then concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (gradient elution with ethyl acetate-hexane 1: 1, then chloroform-methanol 50:1, 40:1, then 30:1) to give compound 8 as a colourless oil (35 mg, 74%). Deprotection of compound 8 (35 mg, 0.04 mmol) was carried out in methanol (3 mL) andpyridiniump-toluenesulfonate (66 mg, 0.26 mmol). The reaction mixture was stirred at room temperature for 18 hours and then concentrated under reduced pressure. The crude product was purified by preparative TLC (chloroform-methanol 30:1, developed twice) to give GM220 (4.1 mg, 14%). GM179

¹H-NMR (500MHz, CDCl₃) £0.88 (3H, d, J= 6.8 Hz), 0.93 (3H, t, J= 7.4 Hz), 0.93 (3H, d, J = 7.1 Hz), 1.17 (3H, d, J = 6.8 Hz), 1.22 (3H, s), 1.30 (IH, m), 1.37 (IH, m), 1.43 (IH, m), 1.55 (IH, m), 1.68 (IH, m), 1.71 (IH, m), 1.75 (3H, s), 1.93 (IH, m), 2.09 (3H, s), 2.23 (IH, m), 2.52 (IH, m), 2.55 (IH, m), 2.62 (IH, dd, J= 15.0, 3.6 Hz), 2.84 (IH, dd, J= 4.2, 2.2 Hz), 2.92 (IH, dd, J = 6.9, 2.2 Hz), 3.40 (IH, m), 3.40 (3H, s), 3.52 (IH, d, J = 11.1 Hz), 3.55 (IH, m), 3.75 (IH, m), 5.09 (IH, d, J = 9.1 Hz), 5.15 (IH, d, J = 10.6 Hz), 5.61 (IH, dd, J= 15.1, 9.4 Hz), 5.65 (IH, m), 5.68 (IH, m), 6.08 (IH, d, J= 10.8 Hz), 6.27 (IH, dd, J= 15.2, 10.8 Hz); (+)-HR-ESIMS m/z 589.3377 [MH-Na]⁺

GM220

¹H NMR (500 MHz, CDCl₃) £0.88 (3H, t, J= 7.4 Hz), 0.89 (3H, d, J= 6.4 Hz), 0.94 (3H, d, J= 7.1 Hz), 1.16 (3H, d, J= 6.8 Hz), 1.22 (3H, s), 1.28 (IH, m), 1.38 (IH, m), 1.41 (IH, m), 1.55 (IH, m), 1.68 (IH, m), 1.70 (IH, m), 1.76 (3H, s), 1.98 (IH, m), 2.10 (3 H, s), 2.24 (IH, m), 2.36 (3H, br s), 2.45 (4H, m), 2.52 (IH, m), 2.55 (IH, dd, J= 14.9, 3.0 Hz), 2.61 (IH, dd, J = 14.9, 3.7 Hz), 2.79 (IH, dd, J = 6.7, 2.0 Hz), 2.98 (IH, dd, J = 5.9, 2.0 Hz), 3.08 (IH, ddd, J= 6.7, 6.7, 2.5 Hz), 3.31 (3H, s), 3.57 (4H, m), 3.75 (IH, m), 4.75 (IH, dd, J= 8.7, 5.9 Hz), 5.09 (IH, d, J= 9.1 Hz), 5.16 (IH, d, J= 10.7 Hz), 5.61 (IH, dd, J= 15.2, 9.3 Hz), 5.66 (IH, m), 5.69 (IH, m), 6.08 (IH, d, J= 10.5 Hz), 6.29 (H-14, IH, dd, J= 14.8, 10.5 Hz); (+)-HR-ESIMS m/z 693.4235 [M+Hj⁺

To a solution of N1523 (23 mg, 0.04 mmol), DMAP (2.6 mg, 0.02 mmol) and triethylamine (6 μL, 0.04 mmol) in anhydrous THF (2 niL) was added chlorotriethylsilane (7 μL, 0.04 mmol). The reaction mixture was stirred at room temperature for 1 hour. Additional chlorotriethylsilane (14 μL, 0.08 mmol) and triethylamine (12 μL, 0.08 mmol) were added twice over a period of 1 hour. The mixture was quenched with water (1 mL). The bulk of THF was removed under reduced pressure. The residue was extracted with ethyl acetate (2 x 5 mL). The organic extracts were combined, dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by automated flash silica gel column chromatography (gradient elution with hexane alone to ethyl acetate-hexane 4:1) to give compound 9 as a colourless oil (16.5 mg, 50%).

To a solution of compound 9 (24 mg, 0.03 mmol), DMAP (2 mg, 0.02 mmol) and triethylamine (8.6 μL, 0.06 mmol) in anhydrous CH₂Cl₂ (2 mL) was added 4- nitrophenylchloroformate (12.5 mg, 0.06 mmol). The reaction mixture was stirred at room temperature for 1 hour. Additional 4-nitrophenylchloroformate (19.5 mg, 0.1 mmol) and triethylamine (14 μL, 0.1 mmol) were added. The mixture was stirred at room temperature for another hour, and then washed with saturated aqueous NaHCO₃ (3 ^ 5 mL). The organic layer was dried with anhydrous MgSO₄, filtered, and then concentrated. To a solution of crude carbonate in anhydrous THF (2 mL) was added l-(2-aminoethyl)pyrrolidine (8 μL, 0.06 mmol) thrice over a period of 1 hour. The mixture was then concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (elution with ethyl acetate-hexane 1:1, then chloroform-methanol 20:1) to give compound 10 (24 mg, 84%). Deprotection of compound 10 (24 mg, 0.026 mmol) was carried out in methanol (2 mL) and pyridinium p-toluenesulfonate (52 mg, 0.208 mmol). The reaction mixture was stirred at room temperature for 18 hours and then concentrated under reduced pressure. The residue was taken into CH₂Cl₂ (5 niL) and washed with saturated aqueous NaHCO₃ (5 mL). The organic layer was dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by preparative TLC (chloroform-methanol 9:1) to give the title compound GM240 (7 mg, 39%).

Compound 9 (R₃, Ri_?-diTES protected N1523)

¹H NMR (500 MHz, CDCl₃) (50.64 (12H, m), 0.88 (3H, d, J= 6.7 Hz), 0.93 (3H, d, J= 7.1 Hz), 0.96 (3H, m), 0.96-0.99 (18H, m), 1.09 (3H, d, J= 6.8 Hz), 1.21 (3H, s), 1.41 (IH, m), i 1.44 (IH, m), 1.47 (2H, m), 1.48 (2H, m), 1.64 (IH, m), 1.70 (3H, s), 2.10 (3H, s), 2.41 (2H, m), 2.48 (IH, dd, J= 13.9, 3.3 Hz), 2.51 (IH, m), 2.78 (2H, br d, J= 6.9 Hz), 3.08 (IH, dd, J= 6.9, 6.9 Hz), 3.61 (IH, m), 3.86 (IH, m), 4.96 (IH, d, J= 10.7 Hz), 5.08 (IH, d, J= 9.0 Hz), 5.63 (IH, m), 5.65 (IH, m), 5.67 (IH, m), 6.09 (IH, d, J= 10.8 Hz), 6.23 (IH, dd, J= 15.0, 10.8 Hz); (+)-HR-ESIMS m/z 803.4745 [M+Na]⁺

GM240

¹H NMR (500 MHz, CDCl₃) <50.88 (3H, d, J= 6.9 Hz), 0.90 (3H, d, J= 8.0 Hz), 0.91 (3H, t, J= 6.9 Hz), 1.14 (3H, d, J= 6.8 Hz), 1.22 (3H, s), 1.32 (IH, m), 1.38 (IH, m), 1.55 (2H, m), 1.61 (2H, m), 1.68 (IH, m), 1.74 (3H, s), 1.82 (4H, m), 2.10 (3H, s), 2.47 (IH, m), 2.51 (IH, m), 2.55 (IH, m), 2.60 (4H, m), 2.63 (IH, m), 2.67 (2H, m), 2.81 (IH, dd, J= 4.7, 2.1 Hz ), 2.84 (IH, dd, J = 7.9, 2.1 Hz), 3.32 (IH, m), 3.33 (2H, m), 3.75 (IH, m), 4.84 (IH, m), 5.09 (IH, d, J = 9.2 Hz), 5.15 (IH, d, J = 10.7 Hz), 5.38 (IH, br s), 5.61 (IH, dd, J = 15.2, 9.4 Hz), 5.68 (IH, dd, J= 15.2, 9.2 Hz), 5.69 (IH, dd, J = 15.0, 8.7 Hz), 6.09 (IH, d, J= 10.8 Hz), 6.31 (IH, dd, J= 15.0, 10.8 Hz); (Hh)-HR-ESIMS m/z 693.4435 [M+H]⁺ GM242 (prepared by deacetylation of N1523)

d, J= 6.7 Hz), 0.93 (3H, d, J= 7.1 Hz), 0.97 (3H, t, J= 7.3 Hz), 1.15 (3H, d, J= 6.8 Hz), 1.26 (IH, m), 1.31 (3H₅ s), 1.39 (IH, m), 1.43 (IH, m), 1.49 (IH, m), 1.52 (2H, m), 1.70 (IH, m), 1.75 (3H, s), 2.49 (IH₅ m), 2.51 (IH, m), 2.54 (IH, m), 2.61 (IH, dd, J= 14.9, 3.8 Hz), 2.88 (IH, dd, J= 5.1, 2.3 Hz), 2.93 (IH, dd, J = 7.7, 2.3 Hz), 3.30 (IH, dd, J= 5.9, 5.1 Hz), 3.62 (IH, m), 3.74 (IH, m), 3.81 (IH, d, J= 9.7 Hz), 5.14 (IH, d₅ J= 10.7 Hz), 5.42 (IH, dd, J= 15.1, 9.9 Hz), 5.69 (IH, dd, J= 15.0, 8.6 Hz), 5.73 (IH, dd, J= 15.1, 9.7 Hz), 6.08 (IH, d, J = 10.8 Hz), 6.32 (IH, dd, J= 15.0, 10.8 Hz); (+)-HR-ESIMS m/z 533.3000 [M+Na]⁺

Scheme 11 Procedure for the Syntheses of R7 and Ri₇ Bisubstituted Macrolides

(A)R₇, Ri₇-Biscarbamate

GM252

43%

To a solution of GM225 (170 mg, 0.25 mnαol), DMAP (15 nig, 0.12 mmol) and triethylamine (35 μL, 0.25 mmol) in anhydrous THF (8 niL) was added clilorotrietliylsilane

(42 μL, 0.25 mmol). The reaction mixture was stirred at room temperature for 0.5 hour. Additional chlorotriethylsilane (42 μL, 0.25 mmol) and triethylamine (35 μL, 0.25 mmol) were introduced. The mixture was stirred at room temperature for another 30 min and then quenched with water (1 mL). THF was removed under reduced pressure and the residue was extracted with ethyl acetate (2 x 10 mL). The organic extracts were combined, dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by preparative TLC (chloroform-methanol 30:1, then 20: 1) to give the compound 11 as a colourless oil (75 mg, 38%).

To a solution of compound 11 (75 mg, 0.10 mmol), DMAP (6 mg, 0.05 mmol) and triethylamine (40 μL, 0.29 mmol) in anhydrous CH₂Cl₂ (5 mL) was added 4-nitrophenyl chloroformate (38 mg, 0.19 mmol). The reaction mixture was stirred at room temperature for 1 hour. Additional 4-nitrophenyl chloroformate (133 mg, 0.66 mmol), triethylamine (78 μL, 0.56 mmol) and DMAP (6 mg, 0.05 mmol) were added portion-wise over a period of 20 hours. The reaction mixture was washed with saturated aqueous NaHCO₃ (3 x 10 mL). The organic layer was dried with anhydrous MgSO₄, filtered and concentrated under reduced pressure to give the crude carbonate.

To a solution of crude carbonate (0.10 mmol) in anhydrous THF (3 mL) was added 4-(2-aminoethyl)-morpholine (25 μl, 0.19 mmol). The reaction mixture was stirred at room temperature for 30 min. Additional 4-(2-aminoethyl)-morpholine (25 μl, 0.19 mmol) was added. The reaction mixture was stirred at room temperature for another 1.5 hours and then concentrated under reduced pressure. The crude product was purified first by silica gel column chromatography (gradient elution with 50% ethyl acetate-hexane to 12.5% ethyl acetate-hexane, then 2% chloroform-methanol to 5% chloroform-methanol) followed by preparative TLC (chloroform-methanol 20:1) to give the triethylsilyl-protected biscarbamate 12 (35 mg, 39%).

To a solution of compound 12 (35 mg, 0.04 mmol) in methanol (3 mL) was added pyridinium p-toluenesulfonate (19 mg, 0.08 mmol). The reaction mixture was stirred at room temperature for 1 day and then concentrated under reduced pressure. The crude product was purified by preparative TLC (chloroform-methanol 15:1, developed thrice) to give GM252 (13 mg, 42%). GM252

¹H NMR (500 MHz, CDCl₃) <50.86 (3H, t, J= 7.5 Hz), 0.87 (3H, d, J= 6.1 Hz), 0.88 (3H, d, J= 6.2 Hz), 1.10 (3H, d, J= 6.8 Hz), 1.25 (3H, s), 1.32 (IH, m), 1.34 (IH, m), 1.36 (IH, m), 1.44 (IH, m), 1.53 (IH, m), 1.64 (IH, m), 1.71 (IH, m), 1.73 (3H, s), 2.47 (8H, m), 2.49 (4H, m), 2.51 (IH, m), 2.54 (IH, m), 2.60 (IH, dd, J= 14.9, 3.6 Hz), 2.65 (IH, m), 2.84 (IH, br d, J= 6.4 Hz), 2.86 (IH, dd, J= 8.2, 1.9 Hz), 3.17 (IH, ddd, J = 6.4, 6.4, 4.1 Hz), 3.30 (4H, m), 3.38 (3 H, s), 3.71 (8H, m), 3.72 (IH, m), 4.50 (IH, dd, J= 7.2, 6.4 Hz), 4.99 (IH, d, J= 9.6 Hz), 5.15 (IH, d, J= 10.6 Hz), 5.27 (2H, br m), 5.60 (IH, dd, J= 15.1, 9.8 Hz), 5.70 (IH, dd, J= 15.1, 9.6 Hz), 5.72 (IH, dd, J= 15.1, 8.4 Hz), 6.09 (IH, d, J= 10.8 Hz), 6.31 (lH, dd, J= 15.1, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) £ 10.9, 11.3, 12.8, 17.5, 17.7, 24.6, 25.6, 30.8, 31.9, 36.4, 38.1, 38.2, 39.3, 40.1, 41.2, 41.7, 54.31, 54.34, 58.4, 58.5, 59.0, 59.4, 60.5, 67.6, 67.7, 70.2, 74.7, 80.3, 83.6, 84.4, 127.1, 127.4, 131.9, 132.8, 137.3, 140.8, 156.4, 156.9, 173.0; (+)-HR-ESIMS m/z 837.5410 [M+H]⁺

(B) R₃, Rπ-biscarbamate

GM287

67%

To a solution of N831 (A) (98 mg, 0.17 mmol), DMAP (10 mg, 0.08 mmol) and triethylamine (48 μL, 0.34 mmol) in anhydrous CH₂Cl₂ (3.5 mL) was added 4-nitrophenyl chloroformate (70 mg, 0.35 mmol). The reaction mixture was stirred at r.t. for 45 min. Additional 4-nitrophenyl chloroformate (140 mg, 0.69 mmol) and trietlrylamme (96 μL, 0.69 mmol) were added portion- wise over a period of 1 hour. The mixture was stirred at room temperature for another 1 hour and then washed with saturated aqueous NaHCO₃ (3 x 4 mL). The organic layer was dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. To a solution of crude carbonate (0.17 mmol) in anhydrous THF (4 niL) was added ammonia solution (25% in water, 324 μL, 17.3 mmol). The reaction mixture was stirred at room temperature for 45 min and then concentrated under reduced pressure. The residue was taken into CH₂Cl₂ (8 mL) and washed with saturated aqueous NaHCO₃ (2 x 5 mL). The organic layer was dried with anhydrous MgSO₄, filtered, and then concentrated under reduced pressure. The crude product was purified by automated silica gel column chromatography (gradient elution with 40% ethyl acetate-hexane to 100% ethyl acetate) to give GM287 (76 mg, 67%).

GM287

¹H NMR (500 MHz, CDCl₃) £0.85 (3H, t, J= 7.3 Hz), 0.85 (3 H, d, J= 6.9 Hz), 0.86^> (3H, d, J= 6.7 Hz), 1.08 (3H, d, J= 6.9 Hz), 1.19 (3H, s), 1.34 (IH, m), 1.42 (IH, m), 1.43 (IH, m), 1.48 (IH, m), 1.61 (IH, m), 1.62 (IH, m), 1.68 (IH, m), 1.70 (3H, s), 2.07 (3H, s), 2.50 (IH, m), 2.53 (IH, dd, J= 14.8, 3.3 Hz), 2.64 (IH, m), 2.66 (IH, dd, J= 14.8, 3.7 Hz), 2.82 (IH, dd, J= 6.1, 2.2 Hz), 2.84 (IH, dd, J= 8.2, 2.2 Hz), 3.14 (IH, ddd, J= 6.4, 6.4, 4.1 Hz), 3.36 (3H, s), 4.46 (IH, dd, J = 7.6, 6.1 Hz), 4.77 (IH, m), 4.97 (IH, d, J= 10.7 Hz), 4.98 (4H, s), 5.04 (IH, d, J= 9.5 Hz), 5.58 (IH, dd, J= 15.2, 9.7 Hz), 5.66 (IH, dd, J= 14.8, 8.5 Hz), 5.67 (IH, dd, J= 15.2, 9.5 Hz), 6.06 (IH, d, J = 10.8 Hz), 6.29 (IH, dd, J= 14.8, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 10.9, 11.2, 12.8, 17.3, 17.6, 22.2, 24.5, 25.5, 27.1, 35.7, 37.8, 40.0, 41.0, 41.4, 59.0, 59.2, 60.4, 72.3, 74.4, 78.2, 79.7, 83.8, 84.4, 126.4, 127.6, 131.8, 132.9, 136.7, 141.5, 157.1, 157.3 170.0, 170.6; (+)~HR-ESIMS m/z 675.3368 [M+Na]⁺ Scheme 12 Syntheses and NMR data of Macrolactams

O

GH/1218, R₁ = H, R₂ = N₃, R₃ = OAc 13%

GM219, R₁ = N₃, R₂ = H, R₃ = OAc 28% N₃, R₃

t

GM238, R = OAc 9% over 2 steps

14, R = OAc

15, R = OH GM269, R = OH 3% over 2 steps

To N831 (A) (0.510 g, 0.901 mmol) was added Millipore water (270 mL) and NaN₃ (7.3 g, 0.11 mol). The reaction mixture was stirred around 50-60 ⁰C until all starting material had reacted as monitored by reverse phase analytical HPLC (isocratic; 1 mL/min; MeCN/H₂O + 0.1% HCOOH; 50:50 over 10 min; Xterra RP18, 5 μm, 4.6x150 mm). The solvent was removed under reduced pressure, and the residue taken up ethyl acetate, filtered and concentrated. The crude product was purified by preparative HPLC (gradient elution; 18 mL/min; MeCN/H₂O + 0.1% HCOOH; 20:80 to 25:75 over 20 min, 25:75 to 30:70 over 30 min, 30:70 to 35:65 over 40 min, 35:65 to 50:50 over 20 min, 50:50 to 60:40 over 10 min and 60:40 to 100:0 over 20 min; Waters NovaPak radial cartridge column, 40x100 mm) to give GM218 (72.9 mg, 13%), GM219 (152.4 mg, 28%) and azide 13 (95.3 mg, 19%).

Synthesis of GM238

GM218 (107 mg, 0.18 mmol) was dissolved in degassed THF (4 mL). Degassed Millipore water (32 μL, 1.78 mmol) was then added, followed by Me₃P (l.M in THF; 900 μL, 0.90 mmol) under an argon atmosphere. The reaction mixture was stirred at room temperature until all starting material had reacted as indicated by TLC (chloroform- methanol 4:1). It was then concentrated under reduced pressure and dried overnight under vacuum. The crude amine 14 was used without further purification or characterization.

Amine 14 (82.9 mg, 0.14 mmol) dissolved in dry CH₂Cl₂ (10 mL), was introduced dropwise to a solution containing 0-benzotriazol-l-yl-N,7V,N'iV'-tetramethyluronium hexafluorophosphate (HBTU) (270 mg, 0.71 mmol) and N, N-diisopropylethylamine (DIPEA) (92 μL, 0.71 mmol) in dry CH₂Cl₂ (35 mL) at room temperature under an argon atmosphere. After all starting material had reacted as indicated by TLC (chloroform- methanol 9:1), the reaction mixture was filtered and concentrated. The crude product was purified by preparative TLC (chloroform-methanol 9:1) to give GM238 (9.3 mg, 9% over 2 steps).

Synthesis of GM269

Azide 13 (95.3 mg, 0.17 mmol) was dissolved in degassed THF (10 mL). Degassed Millipore water (30 μL, 1.67 mmol) was added, followed by Me₃P (1 M in THF; 850 μL, 0.85 mmol) under an argon atmosphere. The reaction mixture was stirred at room temperature until all starting material had reacted as indicated by TLC (chloroform- methanol 9:1). It was then concentrated under reduced pressure and dried overnight under vacuum. The crude amine 15 was used without further purification or characterization.

Amine 15 (40.8 mg, 0.075 mmol) dissolved in dry CH₂Cl₂ (5 mL), was introduced dropwise to a solution containing HBTU (143 mg, 0.38 mmol) and DIPEA (49 μL, 0.38 mmol) in dry CH₂Cl₂ (15 mL) at room temperature under an argon atmosphere. After all starting material had reacted as indicated by TLC (chloroform-mefhanol 9:1), the reaction mixture was filtered and concentrated. The crude product was purified by preparative TLC (chloroform-methanol 9:1) to give GM269 (2.5 mg, 3% over 2 steps).

GM218

¹H-NMR (500MHz, CDCl₃) £0.89 (3H, d, J= 7.0 Hz), 0.90 (3H, t, J= 7.4 Hz), 0.91 (3H, d, J = 6.7 Hz), 1.17 (3H, d, J = 6.8 Hz), 1.19 (3H₅ s), 1.43 (IH, m), 1.46 (IH, m), 1.56 (IH, m), 1.62 (IH, m), 1.65 (2H, m), 1.74 (IH, m), 1.74 (3H, s), 2.09 (3H, s), 2.37 (IH, m), 2.49 (IH, m), 2.51 (2H, m), 2.84 (IH, dd, J = 5.4, 2.3 Hz), 2.92 (IH, dd, J = 8.2, 2.3 Hz), 3.18 (IH, m), 3.32 (IH₅ dd, J= 6.0, 5.4 Hz), 3.40 (3H, s), 3.61 (IH, d, J= 9.2 Hz), 4.01 (IH, m), 5.10 (IH, d, J = 8.0 Hz), 5.49 (IH, dd, J = 15.5, 8.0 Hz)₅ 5.64 (IH, dd, J = 15.5, 8.5 Hz)₅ 5.70 (IH, dd, J = 15.0, 8.4 Hz), 6.00 (IH, d, J= 10.7 Hz), 6.33 (IH, dd, J= 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 11.0, 11.3, 13.3, 17.2, 18.4, 22.2, 23.5, 24.6, 31.1, 34.6, 39.8, 40.2, 43.0, 59.1, 60.4, 61.1, 69.5, 74.3, 75.8, 76.3, 80.9, 84.7, 126.6, 127.0, 130.9, 133.2, 138.0, 139.6, 171.3; (+)-HR-ESIMS m/z 632.3431 [M+Naf

GM219

¹H-NMR (500MHz, CDCl₃) £0.90 (3H, t, J= 7.3 Hz), 0.90 (3H, d, J= 7.0 Hz), 1.09 (3H, s), 1.10 (3H, d, J= 6.3 Hz), 1.15 (3H, d, J= 6.8 Hz), 1.45 (IH, m), 1.48 (IH, m), 1.50 (IH, m), 1.61 (2H, m), 1.62 (IH, m), 1.64 (IH, m), 1.72 (3H, s), 2.06 (3H, s), 2.33 (IH, m), 2.51 (3H, m), 2.88 (IH, dd, J= 5.4, 1.8 Hz), 2.95 (IH, dd, J= 7.9, 1.8 Hz), 3.19 (IH, m), 3.34 (IH, dd, J= 5.4, 5.3 Hz), 3.40 (3H, s), 3.61 (IH, d, J= 9.9 Hz), 3.97 (IH, m), 5.00 (IH, d, J = 7.5 Hz), 5.42 (IH, dd, J= 15.5, 7.5 Hz), 5.47 (IH, dd, J= 15.5, 8.6 Hz), 5.72 (IH, dd, J = 15.1, 7.4 Hz), 5.97 (IH, d, J = 10.5 Hz), 6.28 (IH, dd, J= 15.1, 10.5 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 11.0, 11.3, 13.3, 16.2, 18.8, 22.2, 23.8, 24.6, 31.1, 34.5, 39.6, 40.9, 42.5, 59.0, 60.2, 61.2, 69.4, 74.2, 75.6, 76.5, 80.5, 84.8, 125.8, 126.8, 130.6, 134.3, 138.3, 139.1, 171.1; (+)-HR-ESIMS m/z 632.3368 [M+Na]⁺

GM238

¹H-NMR (500MHz, CDCl₃) £0.89 (3H, d, J= 7.0 Hz), 0.91 (3H, t, J= 7.4 Hz), 0.95 (3H, d, J= 6.6 Hz), 1.14 (3H, d, J= 6.8 Hz), 1.23 (3H, s), 1.41 (3H, m), 1.42 (IH, m), 1.48 (IH, m), 1.64 (IH, m), 1.70 (IH, m), 1.71 (3H, s), 1.95 (IH, d, J = 4.9 Hz), 2.10 (3H s), 2.18 (IH, dd, J= 14.4, 2.9 Hz), 2.28 (IH, m), 2.49 (IH, m), 2.64 (IH, dd, J= 14.4, 3.3 Hz), 2.82 (IH, dd, J= 5.0, 2.2 Hz), 2.92 (IH, dd, J = 8.1, 2.2 Hz), 3.17 (IH, m), 3.32 (IH, m), 3.41 (3H, s), 3.75 (IH, m), 4.31 (IH, dd, J= 11.6, 10.7 Hz), 4.49 (IH, d, J= 10.2 Hz), 5.12 (H-7, IH₅ d, J= 9.5 Hz), 5.52 (IH, d, J= 11.6 Hz), 5.61 (IH, dd, J= 15.3, 9.5 Hz), 5.66 (IH, dd, J= 15.0, 8.4 Hz), 5.71 (IH, dd, J= 15.3, 9.8 Hz), 6.06 (IH, d, J= 10.8 Hz), 6.30 (IH, dd, J = 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) 5 11.0, 11.4, 13.8, 17.5, 18.6, 22.2, 24.7, 25.6, 30.7, 37.0, 39.9, 40.6, 42.7, 43.3, 59.2, 60.2, 60.8, 61.1, 70.5, 74.7, 75.5, 78.6, 80.0, 84.7, 126.0, 127.5, 130.1, 137.3, 142.5, 170.5; (+)-HR-ESIMS m/z 566.3759 [M+H]⁺

GM269

¹H-NMR (500MHz, CDCl₃) 50.89 (3H, d, J= 7.0 Hz), 0.90 (3H, t, J= 7.4 Hz), 0.97 (3H, d, J= 6.6 Hz), 1.15 (3H, d, J= 6.8 Hz), 1.33 (3H, s), 1.36 (3H, m), 1.43 (IH, m), 1.47 (IH, m), 1.65 (IH, m), 1.69 (IH, m), 1.74 (3H, s), 2.29 (IH, dd, J= 14.4, 2.8 Hz), 2.31 (IH, m), 2.49 (IH, m), 2.62 (IH, dd, J= 14.4, 3.4 Hz), 2.82 (IH, dd, J= 5.1, 2.2 Hz), 2.91 (IH, dd, J = 8.2, 2.2 Hz), 3.18 (IH, ddd, J= 6.4, 4.1, 4.0 Hz), 3.33 (IH, dd, J= 6.4, 5.1 Hz), 3.41 (3H, s), 3.74 (IH, m), 3.85 (IH, d, J= 9.5 Hz), 4.29 (IH, dd, J= 10.5, 9.5 Hz), 5.51 (IH, dd, J = 15.2, 9.9 Hz), 5.67 (2H, m), 5.78 (IH, br d, J= 9.5 Hz), 6.05 (IH, d, J= 10.8 Hz), 6.31 (IH, dd, J = 15.0, 10.8 Hz); ¹³C NMR (125 MHz, CDCl₃) δ 11.0, 11.3, 13.9, 17.4, 18.7, 24.7, 25.4, 30.6, 37.4, 39.9, 40.5, 42.6, 43.2, 59.1, 60.2, 60.8, 61.1, 70.7, 74.7, 75.5, 78.3, 84.6, 127.5, 129.9, 130.5, 133.6, 137.3, 139.4, 172.0; (+)-HR-ESIMS m/z 524.3546 [MH-H]⁺

Scheme 13. Syntheses of Epimeric N831 (A) Derivatives

11% over 2 steps

GM161

20%

47%

To a solution of N831 (A) (131 mg, 0.23 mmol) in dry THF (10 mL) was added triethylamine (50 μL, 0.50 mmol) followed by DMAP (14.2 mg, 0.12 mmol). The mixture was then stirred at 0 ⁰C for 15 min. TES-Cl (150 μL, 1.00 mmol) was introduced dropwise to the reaction mixture at 0 ⁰C. After all starting material had reacted as indicated by TLC (ethyl acetate-hexane 1: 1), it was diluted with ethyl acetate, then washed successively with water and brine. The aqueous layer was extracted with CH₂Cl₂, and the combined organic layers dried (MgSO₄), filtered and concentrated. The crude product was purified by flash column chromatography (ethyl acetate-hexane 1:1) to give compound 16 (112 mg, 61%).

To a solution of 16 (108 mg, 0.14 mmol) in dry methanol (4 mL) was added K₂CO₃ (70 mg, 0.51 mmol). After all starting material had reacted as indicated by TLC (ethyl acetate-hexane 1:1), the solvent was removed under reduced pressure. The residue was taken up in ethyl acetate, washed with brine, and the organic layer dried (MgSO₄), filtered and concentrated. The crude product 17 was used for the next step without further purification or characterization. To a cooled (0 ⁰C) solution of crude product 17 (110 mg, 0.15 mmol) in dry CH₂Cl₂ (S mL) was added Dess-Martin periodinane (120 mg, 0.28 mmol). The reaction mixture was then stirred at room temperature overnight. It was worked-up by washing successively with saturated aqueous NaHCO₃ and brine, the organic layer dried (MgSO₄), filtered and concentrated. The crude product was purified by flash column chromatography (ethyl acetate-hexanes 1 :1) to give ketone 18 (11.7 mg, 11%).

To ketone 18 (6.4 mg, 8.53 μmol) was added

(8 mg, 0.03 mmol) in methanol (1 mL). The solution was stirred at room temperature until all starting material had reacted. The solvent was removed under reduced pressure and the crude product purified by preparative TLC (chloroform-methanol 25:1) to give GM161 (0.9 mg, 20%). To a solution of GM161 (4 mg, 7.66 μmol) in dry THF (1.2 mL) was added NaBH₄ (10 mg, 0.26 mmol) at room temperature. After all starting material had reacted as indicated by TLC (ethyl acetate-hexane 9:1), methanol was added. All solvents were then removed under reduced pressure, and the crude product purified by preparative TLC (ethyl acetate-hexane 9:1) to give GM169 (1.9 mg, 47%).

Compound 16 (R₃, R₁₇-diTES protected N831 (A))

¹H NMR (500 MHz, CDCl₃) 50.61 (12H, m), 0.85 (3H, d, J- 6.8 Hz), 0.87 (3H, d, J= 7.4 Hz), 0.88 (3H, t, J = 7.3 Hz), 0.95 (18H, m), 1.07 (3H, d, J = 6.8 Hz), 1.19 (3H, s), 1.32 (IH, m), 1.34 (IH, m), 1.39 (IH, m), 1.42 (2H, m), 1.62 (IH, m), 1.66 (IH, m), 1.68 (3H, s), 2.08 (3H, s), 2.37 (2H, m), 2.45 (IH, dd, J= 13.8, 3.0 Hz), 2.46 (IH, m), 2.67 (IH, dd, J = 6.1, 2.0 Hz), 2.73 (IH, br d, J= 8.1 Hz), 3.10 (IH, dd, J= 6.8, 6.1 Hz), 3.20 (IH, m), 3.39 (3H, s), 3.83 (IH, m), 4.95 (IH, d, J= 10.7 Hz), 5.06 (IH, d, J= 8.6 Hz), 5.63 (2H, m), 5.67 (IH, dd, J= 15.0, 8.3 Hz), 6.06 (IH, d, J= 10.8 Hz), 6.21 (IH, dd, J= 15.0, 10.8 Hz); (+)- HR-ESIMS m/z 817.5201 [M+Na]⁺ GM161

¹H-NMR (500MHz, CDCl₃) £0.88 (3H, d, J= 6.9 Hz), 0.90 (3H, t, J= 7.4 Hz), 0.96 (3H, d, J= 6.7 Hz), 1.10 (IH, m), 1.15 (3H, d, J= 6.8 Hz), 1.39 (3H, s), 1.42 (IH, m), 1.47 (IH, m), 1.57 (IH, m), 1.59 (IH, m), 1.68 (IH, m), 1.72 (IH, m), 1.77 (3H, s), 2.50 (IH, m), 2.55 (IH, dd, J= 15.0, 5.8 Hz), 2.62 (IH, m), 2.64 (IH, m), 2.82 (IH, dd, J= 5.0, 2.1 Hz), 2.91 (IH, dd, J= 8.2, 2.1 Hz), 3.19 (IH, m), 3.32 (IH, m), 3.41 (3H, s), 3.97 (IH, m), 5.11 (IH, d, J= 10.4 Hz), 5.72 (IH, dd, J= 15.1, 8.6 Hz), 6.11 (IH, d, J= 10.8 Hz), 6.33 (IH, dd, J= 15.1, 10.8 Hz), 6.39 (IH, dd, J= 15.5, 10.0 Hz), 6.89 (IH, d, J= 15.5 Hz); (+)-HR-ESIMS

GM169

¹H-NMR (500MHz, CDCl₃) £0.90 (3H, t, J= 7.4 Hz), 0.90 (3H, d, J= 7.0 Hz), 0.92 (3H, d, J = 6.6 Hz), 1.16 (3H, d, J= 6.8 Hz), 1.26 (IH, m), 1.27 (IH, m), 1.39 (3H, s), 1.42 (IH, m), 1.48 (IH, m), 1.63 (IH, m), 1.63 (IH, m), 1.66 (IH, m), 1.77 (3H, s), 1.98 (IH, d, J = 5.8 Hz), 2.49 (IH, m), 2.55 (2H, m), 2.62 (IH, m), 2.82 (IH, dd, J= 5.2, 2.3 Hz), 2.91 (IH, dd, J= 8.2, 2.3 Hz), 3.18 (IH, ddd, J = 6.4, 4.0, 3.9 Hz), 3.31 (IH, m), 3.41 (3H, s), 3.71 (IH, m), 4.02 (IH, m), 5.16 (IH, d, J= 10.5 Hz), 5.53 (IH, ddd, J= 15.4, 9.8, 2.0 Hz), 5.69 (IH, dd, J= 15.0, 8.7 Hz), 5.95 (IH, dd, J= 15.4, 2.4 Hz), 6.11 (IH, d, J= 10.8 Hz), 6.34 (IH, dd, J= 15.0, 10.8 Hz); (+)-HR-ESIMS m/z 547.3247 [M+Na]⁺

Other Embodiments

While the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

What is claimed is:

1. A compound according to Formula I:

or a pharmaceutically acceptable salt or individual diastereomer thereof, wherein Z is O, NH, S, or absent, such that when Z is absent the compound is represented by formula (I^*)

wherein R_d is -NH? or azide;

R₃ is hydrogen, silyl, Ci-C₆ alkyl or substituted Ci-C₆ alkyl, or -C(O)R₅; silyl is R^aR^bR^cSi-, further wherein R^a, R^b, and R^c are the same as or different from each other and each represents methyl, ethyl, i-propyl, t-butyl, or phenyl; R_s is methyl, -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted; R₄ is hydrogen; R₅ and R_5a are each, independently, hydrogen or hydroxyl, or taken together form a carbonyl; or taken together, R₄ and any one Of R₅ or R_5a form a double bond;

R₆ is hydrogen, hydroxyl, Ci-C₆ alkyl or substituted Ci-C₆ alkyl;

R₇ is hydrogen or -C(O)Rt,

R₁ is methyl, -NH₂, Ci-C₆ alkylamino, Ci-Cg dialkylamino, or heterocycle, each of which may be substituted;

Ri₆ and Ri_6a are the same or different from each other and each represents: methyl, hydrogen, or hydroxyl;

Rn is:

(A) -OC(O)R_17b;

(B) sulfonate or substituted sulfonate;

(C) hydroxyl;

(D) silyloxy; or Ri _7a is hydrogen; or taken together Ri₇ and Ri_7a form a carbonyl; Rπb is -NR_jR_k, -OR_m, or -R_n, further wherein,

1) R_j and R_k are the same as or different from each other and each represents: hydrogen; C₂, C₃, C₄, C₅, or C₆ alkyl; C₃, C₄, C5, or C₆ alkenyl; C₂, C₃, C₄, C₅, or C₆ acyl; unsaturated C₃, C₄, C₅, C₆, C₇, or Cs acyl; C5, C₆, C₇, or C₈ aryl; heteroaryl; benzyl; C₁, C₂, C₃, C₄, C₅, or C₆ alkylsulfonyl; benzenesulfonyl; -(CH₂)_x-heteroaryl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or Cg cycloalkenyl; -(CH₂)_y-heterocycle; -(CH₂)_n-NH₂; or methyl, each of which may be substituted; x = 1, 2, or 3; y = 0, 1, 2, or 3; and n = 0, 1, 2, or 3; or

2) -NR_jR_k is bound together to represent a ring, wherein the ring is a non-aromatic heterocyclic ring containing at least one nitrogen atom, wherein any of the atoms in the ring may be substituted;

R_m is C₂, C₃, C₄, C₅, or Cb alkyl; C₃, C₄, C₅, or C₆ alkenyl; C₅, C₆, C₇, or C₈ aryl; heteroaryl; benzyl; -(CH?)_s-heteroaryl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or Cg cycloalkenyl; -(CH₂)_u-heterocycle, -(CH₂)_t-NH₂, or methyl, each of which may be substituted; s = 1, 2, or 3; t = 1, 2, or 3; and u = 0, 1, 2, or 3;

R_n is C₅, C₆, C₇, or C₈ aryl; heteroaryl; C₂, C₃, C₄, C₅, or C₆ alkyl; -(CH₂)_g-heterocycle; -

(CH₂),-C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; -(CH₂)_h-

NH₂; -(CH₂)_j-heteroaryl; or methyl, each of which may be substituted; g = 0, 1, 2, or 3; h =

1, 2, or 3; j = 0, 1, 2, or 3; and i = 0, 1, 2, or 3;

Ri₈ is hydroxyl, halogen, or taken together any one OfR]₇ or Ri_7a and Ri₈ are connected to form a 3, 4, 5, or 6- membered ring containing at least one oxygen atom;

Ri ₉ is hydroxyl, halogen, or -C(O)R₁₁, or

R₁₁ is methyl, aryloxy, -NH₂, Ci-C₆ alkoxy, Ci-C₆ alkylamino, Ci-Ce dialkylamino, or heterocycle, each of which may be substituted, or taken together Ri₈ and Rjg form a double bond or are connected to form a 3, 4, 5, or 6- membered ring containing at least one oxygen atom;

R_2I and R₂i_a are the same or different from each other and each represents: hydrogen, hydroxy, Ci-C₆ alkyl, Ci-C₆ alkoxy, or -OC(O)R_V, or taken together R₂] and R₂i_aform a carbonyl;

R_v is methyl, aryloxy, -NH₂, CrCg alkoxy, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each which may be substituted;

R₂₂ is hydrogen or Ci-C₆ alkyl; and provided that when the above compound is represented by Formula IA:

(IA) wherein, taken together R₄ and any one of R₅ or Rs_a form a double bond and any one of R₂₁ or R₂i_a is hydroxyl, then the other R₂] or R₂i_a is not hydrogen; and wherein when R₄, R₅, and Rs_a are each independently hydrogen and any one of R₂i or R₂i_a is methoxy or hydroxyl, then the other R₂₎ or R₂j_a is not hydrogen.

2. A compound according to Formula II:

or a pharmaceutically acceptable salt or individual diastereomer thereof, wherein Z is O, NH, or S;

R₃ is hydrogen, silyl, C₁-C₆ alkyl or substituted Ci-C₆ alkyl, or -C(O)R₃; further wherein silyl is R^aR^bR^cSi-, wherein R^a, R^b, and R^c are the same as or different from each other and each represents methyl, ethyl, i-propyl, t-butyl, or phenyl; R_s is methyl, -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted;

R₆ is hydrogen, hydroxyl, Ci-C₆ alkyl or substituted Ci-C₆ alkyl; R₇ is hydrogen or -C(O)R_t;

R_t is methyl, -NH₂, Ci-C₆ alkylamino, CrC₆ dialkylamino, or heterocycle, each of which may be substituted; Ri_7b is -NR_jR_k, -OR_m, or -R_n, further wherein,

1) R_j and R_k are the same as or different from each other and each represents: hydrogen; C₂, C₃, C₄, C₅, or C₆ alkyl; C3, C₄, C₅, or C₆ alkenyl; C₂, C₃, C₄, C5, or C₆ acyl; unsaturated C₃, C₄, C₅, C₆, C₇, or C₈ acyl; C₅, C₆, C₇, or C₈ aryl; heteroaryl; benzyl; Ci, C₂, C₃, C₄, C₅, or C₆ alkylsulfonyl; benzenesulfonyl; -(CH₂)_x-heteroaryl; C₃, C₄, C₅, C₆, C₇, or Cg cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkenyl; -(CH₂)_y-heterocycle; -(CH₂)_n-NH₂; or methyl, each of which may be substituted; x = 1, 2, or 3; y = 0, 1, 2, or 3; and n = 0, 1, 2, or 3; or

2) -NR_jR_k is bound together to represent a ring, wherein the ring is a non-aromatic heterocyclic ring containing at least one nitrogen atom, wherein any of the atoms in the ring may be substituted;

R₁₁₁ is C₂, C₃, C₄, C₅, or C₆ alkyl; C₃, C₄, C₅, or C₆ alkenyl; C₅, C₆, C₇, or C₈ aryl; heteroaryl; benzyl; -(CH₂)_s-heteroaryl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkenyl;

-(CH₂)_rNH₂, or methyl, each of which may be substituted; s = 1, 2, or 3; t = 1, 2, or 3; and u = 0, 1, 2, or 3; R_n is C₅, C₆, C₇, or C₈ aryl; heteroaryl; C₂, C₃, C₄, C5, or C₆ allcyl; -(CH₂)_g-lieterocycle; - (CH₂)_I-C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; C₃, C₄, C₅, C₆, C₇, or C₈ cycloalkyl; -(CH₂)_h- NH₂; -(CH₂)_j-heteroaryl; or methyl, each of which may be substituted; g = 0, 1, 2, or 3; h = 1, 2, or 3; j = 0, 1, 2, or 3; and i = 0, 1, 2, or 3.

3. The compound according to claim 2, wherein Z = O.

4. The compound according to claim 2, wherein Z = NH.

5. The compound according to claim 2, wherein Z = S.

6. The compound according to claim 2, wherein R₆ is hydrogen.

7. The compound according to claim 2, wherein R₇ is -C(O)R_t.

8. The compound according to claim 7, wherein R_t is methyl.

9. The compound according to claim 7, wherein R_t is -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle.

10. The compound according to claim 2, wherein R₃ is hydrogen.

11. The compound according to claim 2, wherein R₆ is hydrogen, R₇ is -C(O)CH₃, and R₃ is hydrogen.

12. The compound according to claim 2, wherein R₃ is -C(O)R₅, further wherein R_s is Ci-C₆ allcyl.

13. The compound according to claim 12, wherein R₃ is methyl.

14. The compound according to claim 2, wherein R₇ is hydrogen.

15. The compound according to claim 2, wherein Rj₇b is -NRjR_k, further wherein R_j and R_k are the same as or different from each other and each represents: hydrogen; C₂-C₆ alkyl; C₃-C₆ alkenyl; Cz-C₆ acyl; unsaturated C_3-C₈ acyl; C_5-Cs aryl; heteroaryl; benzyl; Cj-C₆ alkylsulfonyl; benzenesulfonyl; or methyl, each of which may be substituted.

16. The compound according to claim 2, wherein R)₇₁₃ is -NRjR_k, further wherein R_j and R_k are the same as or different from each other and each represents: -(CH₂)_xheteroaryl; C₃- C₈ cycloalkyl; C₃-C₈ cycloalkenyl; heterocycle; -(CH₂)_y-heterocycle; -(CH₂)_n-NH₂; or hydrogen.

17. The compound of claim 2, wherein R₁ -^, is -NR_jR_k, further wherein -NRjR_k is bound together to represent a 3, 4, 5, 6, 7, or 8-membered ring, wherein the ring is a non-aromatic heterocyclic ring containing at least one nitrogen atom, wherein any of the atoms in the ring may be substituted.

18. The compound of claim 17, wherein the ring is a 3-membered ring.

19. The compound of claim 17, wherein the ring is a 4-membered ring.

20. The compound of claim 17, wherein the ring is a 5-membered ring.

21. The compound of claim 17, wherein the ring is a 6-membered ring.

22. The compound of claim 17, wherein the ring is a 7-membered ring.

23. The compound of claim 17, wherein the ring is an 8-membered ring.

24. The compound of claim 17, wherein the ring contains at least two nitrogen atoms.

25. The compound of claim 17, wherein the ring is selected from pyrrolidine, piperidine, azepane, homopiperazine, tetrahydropyridine, tetxahydropyrimidine, morpholine, piperazine, or azocane, each of which may be substituted.

26. The compound of claim 21, wherein the ring is pyrrolidine.

27. The compound of claim 26, wherein pyrrolidine is substituted with hydroxyl or C₁- C₆ alkyl.

28. The compound of claim 27, wherein pyrrolidine is substituted with methylhydroxyl.

29. The compound of claim 21, wherein the ring is piperidine.

30. The compound of claim 29, wherein the piperidine ring is substituted with hydroxyl or pyridine.

31. The compound of claim 21, wherein the ring is 1,2,3,6-tetrahydropyridine or 1,2,5,6- tetrahydropyridine.

32. The compound of claim 31 , wherein the ring is substituted with pyridine.

33. The compound of claim 21, wherein the ring is 2,4,5,6-tetrahydropyrimidine.

34. The compound of claim 21, wherein the ring is morpholine.

35. The compound of claim 21, wherein the ring is piperazine.

36. The compound of claim 35, wherein piperazine is substituted with one Of C₃-C₈ cycloalkyl, Ci-C₆ alkyl, heteroaryl, -C(O)Rc, C5-C8 aryl, or hydroxyl.

37. The compound of claim 36, wherein piperazine is substituted with -C(O)R_e, wherein Rc is Ci-C₆ alkyl.

38. The compound of claim 37, wherein R_c is ethyl.

39. The compound of claim 36, wherein piperazine is substituted with pyridine, phenol, pyrimidine, phenyl, or cyclohexyl.

40. The compound of claim 36, wherein piperazine is substituted with Cj ~C<_> alkyl, further wherein C]-C₆ alkyl is substituted with hydroxyl, -NH₂, Ci-C₆ alkylamino, or Ci-C₆ dialkylamino.

41. The compound according to claim 22, wherein the ring is azepane or homopiperazine .

42. The compound of claim 16, wherein any one of R₁- or R_k is selected from -(CH₂)_X- heteroaryl; C₃-C₈ cycloalkyl; -(CH₂)_y-heterocycle, or -(CH₂)_n-NH₂ and the other Rj or R_k is selected from hydrogen or methyl.

43. The compound of claim 42, wherein the other of R_j or R^ is hydrogen.

44. The compound of claim 42, wherein the other of R_j or R]_cis methyl.

45. The compound of claim 42, wherein any one of Rj or R_k is selected from -(CEb)_x- heteroaryl.

46. The compound of claim 45, wherein x = 1.

47. The compound of claim 45, wherein x = 2.

48. The compound of claim 45, wherein x = 3.

49. The compound of claim 45, wherein heteroaryl is pyridine.

50. The compound according to claim 42, wherein any one of Rj or Rk is C₃-C₈ cycloalkyl.

51. The compound according to claim 50, wherein C₃-C₈ cycloalkyl is cyclohexyl.

52. The compound according to claim 42, wherein any one of R_j or R_k is -(CH₂)_y- heterocycle and the other of R_j or R_k is hydrogen.

53. The compound according to claim 42, wherein heterocycle is a 6-membered ring.

54. The compound according to claim 52, wherein y =0.

55. The compound according to claim 52, wherein y = 1.

56. The compound according to claim 52, wherein y = 2.

57. The compound according to claim 52, wherein y = 3.

58. The compound according to claim 52, wherein heterocycle is selected from pyrrolidine, piperidine, morpholine, pyridine, or piperazine, each of which may be substituted.

59. The compound according to claim 58, wherein heterocycle is pyrrolidine.

60. The compound according to claim 58, wherein heterocycle is pyridine.

61. The compound according to claim 58, wherein heterocycle is piperidine.

62. The compound according to claim 58, wherein heterocycle is morpholine.

63. The compound according to claim 58, wherein heterocycle is piperazine.

64. The compound according to claim 63, wherein piperazine is substituted with methyl.

65. The compound according to claim 42, wherein any one of R_j or R_k is -(CH₂)_n-NH₂.

66. The compound of claim 65, wherein the other of Rj or R_k is hydrogen.

67. The compound of claim 65, wherein the other of Rj or R_k is methyl.

68. The compound according to claim 65, wherein n = 1.

69. The compound according to claim 65, wherein n = 2.

70. The compound according to claim 65, wherein n = 3.

71. The compound according to claim 15, wherein R_j and R_k are both methyl.

72. The compound according to claim 15, wherein R_j and R_k are both hydrogen.

73. The compound according to claim 2, wherein R₁₇₁, is -OR_m.

R_m is C₂-Cg alkyl; C₃-C₆ alkenyl; C₅-Cg aryl; heteroaryl; benzyl; -(CH₂)_s-heteroaryl; C₃-C₈ cycloalkyl; C₃-C₈ cycloalkenyl; -(CH₂)_u-heterocycle, -(CH₂)_t-NH₂, or methyl, each of which may be substituted; s = 1, 2, or 3; t = 1, 2, or 3; and u = 0, 1, 2, or 3.

74. The compound according to claim 73, wherein R₁₁₁ is

C₂-C₆ alkyl; C₃-C₆ alkenyl; C₅-C₈ aryl; heteroaryl; or methyl, each of which may be substituted.

75. The compound according to claim 73, wherein R_m is

-(CH₂)_s-aryl; -(CH₂)_s-heteroaryl; C₃-C₈ cycloalkyl; C₃-C₈ cycloalkenyl; -(CH₂)_U- heterocycle; or -(CH₂)_t-amino, each of which may be substituted.

76. The compound according to claim 74, wherein R_m is C₅-C₈ aryl.

77. The compound according to claim 76, wherein C₅-C₈ aryl is phenyl.

78. The compound according to claim 77, wherein phenyl is substituted with -NO₂.

79. The compound according to claim 2, wherein R] _7b is R_n, further wherein

R_n is C₅-C₈ aryl; heteroaryl; C₂-C₆ alkyl; -(CH₂)_g-heterocycle; -(CH₂)_I-C₃-C₈ cycloalkyl; C₃-C₈ cycloalkyl; -(CH₂)_I1-NH₂; -(CH₂)_j-heteroaryl; or methyl, each of which may be substituted; g = 0, 1, 2, or 3; h = 1, 2, or 3; j = 0, 1, 2, or 3; and i = 0, 1, 2, or 3.

80. The compound according to claim 79, wherein R_n is C₅-C₈ aryl or heteroaryl, each of which may be substituted.

81. The compound according to claim 80, wherein R_n is phenyl or pyridine.

82. The compound according to claim 79, wherein R_π is

Ci-C₆ alkyl; -(CH₂)_g-heterocycle; -(CH₂)J-C₃-C₈ cycloalkyl; C₃-C₈ cycloalkyl; -(CH₂)_h-NH₂; or -(CH₂)_j-heteroaryl, each of which may be substituted.

83. The compound according to claim 82, wherein R_n is selected from Ci-C₆ alkyl, - (CH₂)_h-NH₂, or C₃-C₈ cycloalkyl, each which may be substituted.

84. The compound according to claim 83, wherein R_n is methyl or cyclopropyl.

85. The compound according to claim 82, wherein R_n is -(CH₂)I₁-NH₂.

86. The compound according to claim 85, wherein h = 1.

87. A compound according to Formula III:

a pharmaceutically acceptable salt or individual diastereomer thereof, wherein

Z is O, NH, or S;

R₃ is hydrogen, silyl, or -C(O)R₈; silyl is R^aR^bR^cSi-, further wherein R^a, R^b, and R^c are the same as or different from each other and each represents methyl, ethyl, i-propyl, t-butyl, or phenyl;

R₈ is methyl, -NH₂, allcylamino, dialkylamino, or heterocycle, each which may be substituted;

R₇ is hydrogen or -C(O)CH₃;

Ri₆ and Ri_6a are the same or different from each other and each represents: methyl, hydrogen, or hydroxyl; Rn is hydroxyl, silyloxy; or taken together Rn and R_17a form a carbonyl; or Ri_7a is hydrogen; Ri ₈ is hydroxyl or halogen, or taken together Rn and R₁₈ are connected to form a 3, 4, 5, or 6-membered ring containing at' least one oxygen atom; Ri₉ is hydroxyl, halogen, or -C(O)R_U; or taken together Ri₈ and R₁₉ are connected to form a 3, 4, 5, or 6-membered ring containing at least one oxygen atom;

R₁₁ is methyl, aryloxy, -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted;

R₂₁ and R₂] _a are the same or different from each other and each represents: hydrogen, hydroxy, Ci-C₆ alkyl, Ci-C₆ alkoxy, or -OC(O)R_V, or taken together R₂₁ and R_{2 la} form a carbonyl; R₂₂ is hydrogen or Ci-C₆ alkyl;

R_v is methyl, aryloxy, -NH₂, Ci-C₆ alkylamino, Ci-C₆ dialkylamino, or heterocycle, each of which may be substituted; and provided that when the above compound is represented by Formula IIIA:

(IIIA) if any one Of R₂₁ or R₂i_a is methoxy or hydroxyl, then the other R₂₁ or R₂i_a is not hydrogen.

?. The compound according to claim 87, wherein Z = O.

89. The compound according to claim 87, wherein Z = NH.

90. The compound according to claim 87, wherein Z = S.

91. The compound according to claim 87, wherein R₃ is -C(O)CH₃.

92. The compound according to claim 87, wherein R₃ is hydrogen.

93. The compound according to claim 87, wherein R₇ is -C(O)CH₃.

94. The compound according to claim 87, wherein R₇ is hydrogen.

95. The compound according to claim 87, wherein R₁₈ is a halogen.

96. The compound according to claim 87, wherein Ri₈ is hromide, chloride, or iodide.

97. The compound according to claim 87, wherein R₁₉ is halogen.

98. The compound according to claim 87, wherein R₁₉ is hromide, chloride, or iodide.

99. The compound according to claim 87, wherein R₁₉ is -C(O)R_U.

100. The compound according to claim 99, wherein R₁₁ is aryloxy, -NH₂, Ci-Cg alkylamino, Ci-C₆ dialkylamino, or heterocycle.

101. The compound according to claim 87, wherein R₂₁ is Ci-C₆ allcoxy

102. The compound according to claim 101, wherein R₂₁ is methoxy.

103. The compound according to claim 87, wherein any one of R₂i or R₂i_a is -OC(O)R_V and the other is hydrogen.

104. The compound according to claim 103, wherein R_v is -NH₂, Ci-C₆ alkylamino, C₁- C₆ dialkylamino, or heterocycle.

105. The compound according to claim 87, wherein one of R₁₇ or Ri _7a is hydroxyl and the other is hydrogen.

106. The compound according to claim 87, wherein Rn and Rj₈ are connected to form a 3, 4, 5, or 6-memhered ring containing at least one oxygen atom.

107. The compound according to claim 106, wherein the ring is a cyclic carbonate.

108. The compound according to claim 106, wherein the ring is an epoxide.

109. The compound according to claim 108, wherein R₁₉ is -C(O)R₁₁.

110. The compound according to claim 109, wherein R₁₁ is aryloxy, -NH₂, alkylamino, diallcylamino, or heterocycle.

111. The compound according to claim 108, wherein R₁₉ is hydroxyl.

112. The compound according to claim 87, wherein Rn and Ri₈ are each hydroxyl.

113. The compound according to claim 87, wherein Ri₇ and Ri ₉ are each hydroxyl.

114. The compound according to claim 87, wherein, taken together, R]₇ and Ri_7a form a carbonyl.

115. The compound according to claim 87, wherein, taken together, R₂₁ and R_{2 ]a} form a carbonyl.

116. The compound according to claim 87, wherein any one of R₂₁ or R₂i_a is methyl and the other is hydroxyl.

117. The compound according to claim 87, wherein R₇ is -C(O)CH₃, R₃ is hydrogen, and R₂₁ is methoxy.

118. The compound according to claim 87, wherein R₂₂ is hydrogen or Ci -Cβ alkyl.

119. A compound according to Formula IV:

or a pharmaceutically acceptable salt or individual diastereomer thereof, whei'ein

Z is O, NH, or S;

R₃ is hydrogen, Ci-Cs alkyl or substituted Ci -C<₅ alkyl; R₄ is hydrogen;

R₅ and Rs_a are each independently hydrogen or hydroxyl, or taken together form a carbonyl; or taken together, R₄ and any one Of R₅ or R_5a form a double bond; R₆ is hydrogen, hydroxyl, C]-Cc alkyl or substituted CpCe alkyl; taken together Rj₈ and R]₉ form a double bond or are connected to form a 3, 4, 5, or 6- membered ring containing at least one oxygen atom;

R₂₁ and R₂ i_a are the same or different from each other and each represents: hydrogen, hydroxy, or Ci-C₆ alkoxy, or taken together R₂i and Reform a carbonyl; and provided that when the above compound is represented by Formula IVA:

(IVA) if any of one R₂i or R₂i_a is hydroxyl, then the other R₂i or R₂i_a is not hydrogen

120. The compound according to claim 119, wherein Z = O.

121. The compound according to claim 119, wherein Z = NH.

122. The compound according to claim 119, wherein Z = S.

123. The compound according to claim 119, wherein R₃ is hydrogen.

124. The compound according to claim 119, wherein R₆ is hydroxyl.

125. The compound according to claim 119, wherein R₆ is hydrogen.

126. The compound according to claim 119, wherein R₃ is hydrogen and R₆ is hydroxyl.

127. The compound according to claim 119, wherein any one of R₂] or R₂i_a is hydroxyl.

128. The compound according to claim 127, wherein the other OfR_2I or R_2Ia is hydrogen.

129. The compound according to claim 119, wherein any one of R₅ or R_5a taken together with R₄ forms a double bond.

130. The compound according to claim 119, wherein, taken together, R₅ and Rs₃ foπn a carbonyl.

131. The compound according to claim 119, wherein, taken together, Ri₈ and R₁9 form a double bond.

132. The compound according to claim 119, wherein any one of R₅ or R_5a and R₄ are each hydroxyl.

133. A method of treating an immune-related disorder comprising administering to a patient in need of such treatment a composition comprising a carrier and a non-toxic therapeutically effective amount of a macrolide compound, selected from Formula I₅ I\ II, III, IV or a pharmaceutically acceptable salt or hydrate thereof.

134. The method of claim 133, wherein the immune-related disorder is an autoimmune disease selected from a connective tissue disease, a neuromuscular disease, an endocrine disease, a gastrointestinal disease, an autoimmune skin disease, a vasculitis syndrome, a hematologic autoimmune disease, and uveitis.

135. The method of claim 134, wherein the autoimmune disease is selected from systemic lupus erythematosus (SLE), systemic sclerosis (scleroderma), Sjogren's syndrome, multiple sclerosis (MS), myasthenis gravis, Guillain-Barre syndrome, Hashimoto's thyroiditis, Graves' disease, insulin-dependent (Type 1) diabetes, inflammatory bowel disease, Crohn's disease, ulcerative colitis, and psoriatic arthritis.

136. The method of claim 133, wherein the immune-related disorder is an inflammatory disorder selected from Alzheimer's disease, asthma, atopic allergy, allergy, bronchial asthma, diabetic retinopathy, eczema, glomerulonephritis, graft vs. host disease, hemolytic anemias, sepsis, stroke, vasculitis, and ventilator induced lung injury.

137. A method of alleviating a symptom associated with an immune-related disorder comprising administering to a patient in need of such treatment a composition comprising a carrier and a non-toxic, therapeutically effective amount of a macrolide compound according to Formula I, I', II, III, IV, or a pharmaceutically acceptable salt or hydrate thereof.

138. The method of claim 137, wherein the immune-related disorder is an autoimmune disease selected from a connective tissue disease, a neuromuscular disease, an endocrine disease, a gastrointestinal disease, an autoimmune skin disease, a vasculitis syndrome, a hematologic autoimmune disease, and uveitis.

139. The method of claim 138, wherein the autoimmune disease is selected from systemic lupus erythematosus (SLE), systemic sclerosis (scleroderma), Sjogren's syndrome, multiple sclerosis (MS), myasthenis gravis, Guillain-Barre syndrome, Hashimoto's thyroiditis, Graves' disease, insulin-dependent (Type 1) diabetes, inflammatory bowel disease, Crohn's disease, ulcerative colitis, and psoriatic arthritis.

140. The method of claim 137, wherein the immune-related disorder is an inflammatory disorder selected from Alzheimer's disease, asthma, atopic allergy, allergy, bronchial asthma, diabetic retinopathy, eczema, glomerulonephritis, graft vs. host disease, hemolytic anemias , sepsis, stroke, vasculitis, and ventilator induced lung injury.

141. The method of claim 137, wherein the macrolide is a racemic mixture of a macrolide compound according to Formula I, I', II, III, or IV.

142. The method according to claim 133 or 137, wherein the macrolide is an enantiomerically pure form of a macrolide compound according to Formula I, P, II, III, or IV.

143. The method according to claim 133 or 137, wherein the macrolide is administered in combination with a second agent used to treat an immune-related disorder.

144. The method of claim 143, wherein the second agent used to treat an immune- related disorder selected from methotrexate, cyclosporin A, tacrolimus, corticosteroids, statins, interferon beta, nonsteroidal anti-inflammatory drugs (NSAIDs), and the disease- modifying anti-rheumatic drugs (DMARDs).

145. The method of claim 144, wherein the second agent is cyclosporin A, further wherein the cyclosporin A is cyclosporin microemulsion.

146. A method of suppressing an immune response associated with organ or tissue transplantation comprising administering to a patient in need of such treatment a composition comprising a carrier and a non-toxic, therapeutically effective amount of a macrolide compound according to Forumulae I, I', II, III, IV, or a pharmaceutically acceptable salt or hydrate thereof.

147. The method of claim 146, wherein the macrolide is administered in combination with a second agent used to suppress an immune response associated with organ or tissue transplantation.

148. The method of claim 147, wherein the second agent used to suppress an immune response associated with organ or tissue transplantation is selected from methotrexate, cyclosporin A, cyclosporin microemulsion, tacrolimus, corticosteroids and statins.

149. The method of claim 148, wherein the second agent is cyclosporin A, further wherein the cyclosporin A is cyclosporin microemulsion.

150. The method of claim 146, wherein the macrolide is an enantiomerically pure form of a macrolide compound according to Formula I, V, II, III, or IV.

151. The method of claim 146, wherein the macrolide is administered to said patient at a time selected from prior to said organ or tissue transplantation, during said organ or tissue transplantation, after said organ or tissue transplantation, and combinations thereof.

152. A method of inliibiting tumor growth comprising administering to a patient in need of such treatment a composition comprising a carrier and a non-toxic therapeutically effective amount of a macrolide compound, wherein the macrolide compound is a macrolide compound according to Formula I, I', II, III, IV, or a pharmaceutically acceptable salt or hydrate thereof.

153. The method according to claims 133 or 137, wherein the macrolide compound is administered in combination with a second agent used to inhibit tumor growth.