AU707991B2

AU707991B2 - Triterpene derivatives with immunosuppressant activity

Info

Publication number: AU707991B2
Application number: AU76020/96A
Authority: AU
Inventors: Robert K Baker; Jianming Bao; Frank Kayser; William H Parsons; Kathleen M. Rupprecht
Original assignee: Merck and Co Inc
Current assignee: Merck and Co Inc
Priority date: 1995-10-31
Filing date: 1996-10-28
Publication date: 1999-07-29
Anticipated expiration: 2016-10-28
Also published as: JP2002503201A; WO1997016182A1; EP0858332A4; EP0858332A1; CA2237607A1; AU7602096A

Description

WO 97/16182 PCT/US96/17482 -1- TITLE OF THE INVENTION TRITERPENE DERIVATIVES WITH IMMUNOSUPPRESSANT

ACTIVITY

BACKGROUND OF THE INVENTION Immunoregulatory abnormalities have been shown to exist in a wide variety of "autoimmune" and chronic inflammatory diseases, including systemic lupus erythematosis, chronic rheumatoid arthritis, type I and II diabetes mellitus, inflammatory bowel disease, biliary cirrhosis, uveitis, multiple sclerosis and other disorders such as Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, psoriasis, ichthyosis, Graves ophthalmopathy and asthma.

Although the underlying pathogenesis of each of these conditions may be quite different, they have in common the appearance of a variety of autoantibodies and self-reactive lymphocytes. Such selfreactivity may be due, in part, to a loss of the homeostatic controls under which the normal immune system operates.

Similarly, following a bone-marrow or an organ transplantation, the host lymphocytes recognize the foreign tissue antigens and begin to produce antibodies which lead to graft rejection.

One end result of an autoimmune or a rejection process is tissue destruction caused by inflammatory cells and the mediators they release. Anti-inflammatory agents such as NSAID's act principally by blocking the effect or secretion of these mediators but do nothing to modify the immunologic basis of the disease. On the other hand, cytotoxic agents, such as cyclophosphamide, act in such a nonspecific fashion that both the normal and autoimmune responses are shut off.

Indeed, patients treated with such nonspecific immunosuppressive agents are as likely to succumb from infection as they are from their autoimmune disease.

Cyclosporin A (CsA), which was approved by the US FDA in 1983 is currently the leading drug used to prevent rejection of transplanted organs. In 1993, FK-506 (Prograf) was approved by the US FDA for the prevention of rejection in liver transplantation. CsA F WO 97/16182 PCT/US96/17482 -2and FK-506 act by inhibiting the body's immune system from mobilizing its vast arsenal of natural protecting agents to reject the transplant's foreign protein. In 1994, CsA was approved by the US FDA for the treatment of severe psoriasis and has been approved by European regulatory agencies for the treatment of atopic dermatitis.

Though they are effective in fighting transplant rejection, CsA and FK- 506 are known to cause several undesirable side effects including nephrotoxicity, neurotoxicity, and gastrointestinal discomfort.

Newer, safer drugs exhibiting less side effects are constantly being searched for in the field.

Four active components of Spachea correa were recently identified which inhibit thymidine uptake of T cells and are useful as immunosuppressive agents in animals, including man.

H H Formula 1(a) b is a single H f bond and R is OAc b CH O Formula 1(b) b is a double H H bond and R is OAc S OH 'COOCH3 O CHs CH3 OAc Formula 1(c) b is a single 0 G OAc bond and R is OH CH1 OAc OAc 3 OR Formula 1(d) b is a double bond and R is OH These compounds are useful as immunosuppressive agents in animals, including man. The present invention describes newly developed immunosuppressive compounds derived from the compounds described in Formulae l(a) through 1(d) and which have the relative stereochemistry depicted above.

my- WO 97/16182 PCT/US96/17482 -3- SUMMARY OF THE INVENTION This invention relates to a class of triterpene derivatives of the general structural Formula I

R

29 a R 29 b C '2 H18 22 O 2 1 H 9 H13 OH O 2

CH

3 2 1 _16 O 3 b 10 258 15 OAc ;iOAc 23 a R 4

I

are useful as immunosuppressives.

As an immunosuppressive, the compounds of this invention are useful in the treatment of autoimmune diseases, the prevention of rejection of foreign organ transplants and/or related afflictions, diseases and illnesses. Also within the scope of this invention are pharmaceutical formulations comprising a compound of Formula I and a pharmaceutical carrier, as well as, pharmaceutical formulations comprising a compound of Formula I, a second immunosuppressive compound and a pharmaceutical carrier.

DETAILED DESCRIPTION OF THE INVENTION A. Scope of the Invention The present invention is related to compounds of formula I, including but not limited to those specified in the examples, which are useful in a mammalian subject for the treatment and prevention of the resistance by transplantation of organs or tissue, graft-versus-host diseases brought about by medulla ossium transplantation; rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes uveitis, juvenileonset or recent-onset diabetes mellitus, posterior uveitis, allergic WO 97/16182 PCT/US96/17482 -4encephalomyelitis, glomerulonephritis, infectious diseases caused by pathogenic microorganisms, inflammatory and hyperproliferative skin diseases, psoriasis, atopical dermatitis, contact dermatitis, eczematous dermatitises, seborrhoeis dermatitis, Lichen planus, Pemphigus, bullous pemphigoid, Epidermolysis bullosa, urticaria, angioedemas, vasculitides, erythemas, cutaneous eosinophilias, Lupus erythematosus, acne, Alopecia areata, keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic keratitis, conical cornea, dystrophia epithelialis comeae, corneal leukoma, ocular pemphigus, Mooren's ulcer, Scleritis, Graves' opthalmopathy, Vogt- Koyanagi-Harada syndrome, sarcoidosis, etc.; pollen allergies, reversible obstructive airway disease, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma and dust asthma, chronic or inveterate asthma, late asthma and airway hyper-responsiveness, bronchitis, gastric ulcers, vascular damage caused by ischemic diseases and thrombosis, ischemic bowel diseases, inflammatory bowel diseases, necrotizing enterocolitis, intestinal lesions associated with thermal bums and leukotriene B4-mediated diseases, Coeliac diseases, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease, ulcerative colitis, migraine, rhinitis, eczema, interstitial nephritis, Good-pasture's syndrome, hemolytic-uremic syndrome, diabetic nephropathy, multiple myositis, Guillain-Barre syndrome, Meniere's disease, polyneuritis, multiple neuritis, mononeuritis, radiculopathy, hyperthyroidism, Basedow's disease, pure red cell aplasia, aplastic anemia, hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia, anerythroplasia, osteoporosis, sarcoidosis, fibroid lung, idiopathic interstitial pneumonia, dermatomyositis, leukoderma vulgaris, ichthyosis vulgaris, photoallergic sensitivity, cutaneous T cell lymphoma, arteriosclerosis, atherosclerosis, aortitis syndrome, polyarteritis nodosa, myocardosis, scleroderma, Wegener's granuloma, Sjogren's syndrome, adiposis, eosinophilic fascitis, lesions of gingiva, periodontium, alveolar bone, substantia ossea dentis, glomerulonephritis, male pattern alopecia or alopecia senilis by preventing epilation or providing hair germination WO 97/16182 PCT/US96/17482 and/or promoting hair generation and hair growth; muscular dystrophy; Pyoderma and Sezary's syndrome, Addison's disease, ischemiareperfusion injury of organs which occurs upon preservation, transplantation or ischemic disease, for example, thrombosis and cardiac infraction, endotoxin-shock, pseudomembranous colitis, colitis caused by drug or radiation, ischemic acute renal insufficiency, chronic renal insufficiency, toxinosis caused by lung-oxygen or drug, for example, paracort and bleomycins), lung cancer, pulmonary emphysema, cataracta, siderosis, retinitis, pigmentosa, senile macular degeneration, vitreal scarring, corneal alkali bum; dermatitis erythema multiforme, linear IgA ballous dermatitis and cement dermatitis, gingivitis, periodontitis, sepsis, pancreatitis, diseases caused by environmental pollution, aging, carcinogenis, metastasis of carcinoma and hypobaropathy; disease caused by histamine or leukotriene-C4 release; Behcet's disease, autoimmune hepatitis, primary biliary cirrhosis sclerosing cholangitis), partial liver resection, acute liver necrosis, necrosis caused by toxin, viral hepatitis, shock, or anoxia, B-virus hepatitis, non-A/non-B hepatitis, cirrhosis, alcoholic cirrhosis, hepatic failure, fulminant hepatic failure, late-onset hepatic failure, "acute-onchronic" liver failure, augmention of chemotherapeutic effect, preventing or treating activity of cytomegalovirus infection, HCMV infection, and antiinflammatory activity; and treatment of immunodepression or a disorder involving immunodepression, including AIDS, cancer, senile dementia, trauma, chronic bacterial infection, and certain central nervous system disorders.

More particularly this invention relates to compounds of the general structural formula I:

C

11 2 1 H9 or a pharmaceutically acceptable salt, crystal form, or hydrate, wherein: X is: O, S, NH or H and R1; a is: a single bond, or a double bond when R 4 is absent; b and c are S. 0@ a

S..

independently: n is: m is: r is: o s is:

R

1 is: a) ooo S 0 0060 20 a single bond or a double bond; 1 to 4; 1 to4; 0or1; 0or1; H, or

(C

1

-C

6 )-alkyl, wherein alkyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, I,

(C

1

-C

6 )-alkoxy, vinyl, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 alkyl, C02C 1

-C

6 -alkyl, CONRlaR2, NRlaR2, NRlaCOC 1

-C

6 -alkyl, aryl, wherein aryl is defined as phenyl or naphthyl, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1

-C

6 )-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO,

CO

2 H, COC1-C 6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONRlaR2, NRlaR2, NRlaCOC 1

C

6 -alkyl and any two of adjacent substituents can be joined to form a 6or 7- membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, heteroaryl, wherein heteroaryl is defined as a or 6-membered ring substituted with one and two heteroatoms selected from O, S, N, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, I, (C1-C6)alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 -alkyl, CO 2

C

1

-C

6 -alkyl, CONRlaR2, NRlaR 2 NRlaCOC 1 -C6-alkyl, any two adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring: [N:\LIBAA]01587.doc:TAB Ria and R 2 are independently: a) (0 1

-C

6 )-alkyl, or b) aryl, wherein aryl is phenyl or naphthyl;

R

3 is: a) 1

-C

6 )-alkyl, alkyl as defined above; b) -(0 1

-C

6 )-alkenyl, wherein alkenyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC,-C 6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONRlaR2, NRlaR 2 NRlaCOC 1

-C

6 -alkyl, aryl as defined above, and heteroaryl as defined above; C) -(Cl-C 6 )-alkynyl, wherein alkynyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONRlaR 2 NRlaR 2 NRlaCOC-C 6 -alkyl, aryl as defined above, and heteroaryl as defined above, d) -aryl, aryl as defined above, or e) -heteroaryl, heteroaryl as defined above; R4 is: a) absent and a is a double bond; 66 b -H, c) -OH, d) =0, e) -O[(C=O)Or~sCi-Cioalkyl, alkyl as defined above, 0 f) 0

(C=

0 )Or]sC 2 -Cloalkenyl, as defined above, g) O[1(C=%Or]sC2-C 6 -alkynyl, alkynyl as defined above, h) O0[(C=O)Orls(C 3

-C

7 )-cycloalkyl, .0025 i) O0[(C=O)Orlsaryl, aryl as defined above, j) O0[(C=O)Orjsheteroaryl, heteroaryl as defined above, k) 0 0C 2 n H h te o r l e e o r s d f n d a o e 00 -O(CH 2 )nO(CH 2 )mhraryl, eraryl as defined above, 0000 m) -OC(=O)NRlaR 2 n) -0S0 2

R

3 or o -NRlaR2;

R

29 a and R29b are independently: a) -H, b) -(CH2)s0[I(C0)Or]sCl-ClO-alkyl, alkyl as defined above, c) -(CH2)sO[(C0)Or]sC 2 -ClO-alkenyl, alkenyl as defined above, d) -(CH2)sO[(C0)Or]sC 2

-C

6 alkynyl, alkynyl as defined above, e) -(CH2)s-O[(C=O)Orls(C 3

-C

7 )-cycloalkyl, f) -(CH2)sO[(C0)Orlsaryl, aryl as defined above, g) -(CH2)$-O[(C=O)OrJsheteroaryl, heteroaryl as defined above, Sh) -(CH2)s-O(CH 2 )nO(CH 2 )mheteroaryl, heteroaryl as defined above, [N:\LIBAA1OI 587.doc:TAB i) -(CH2)s-O(CH2)nO(CH 2 )maryl, aryl as defined above, j) -(CH 2 )s-OC(=O)NRlaR 2 k) -(CH 2 )s-OS0 2

R

3 I) -(Ci-C 6 )-alkyl, alkyl as defined above, or m) -(C 2

-C

6 )-alkenyl, alkenyl as defined above;

R

2 9a and R29b together are =0 or =C(C 1

-C

10 -alkyl) 2 alkyl as defined above.

An embodiment of the invention are the compounds of Formula I R29 R29b R\ RFe se ::06 0 I 4 0000 0 0 s o 00 @00.

S.

00 0 or a pharmaceutically acceptable salt, crystal form, or hydrate, wherein: X is: 0, S, or NH; a is: a single bond: b and c are independently: a single bond or a double bond; n is: 1 to 4; 15 m is: 1 to 4; ris: Oor 1; sis: 0or1;

R

1 is: a) 20 0 0*00 20 25 H, or (C1-C6)-alkyl, wherein alkyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (Cl-C 6 )-alkoxy, vinyl, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 alkyl, C02C0-C 6 -alkyl, CONR 1 aR2, NRlaR2, NRlaCOC -C 6 -alkyl, aryl, wherein aryl is defined as phenyl or naphthyl, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, I, (C 1

-C

6 )-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO,

CO

2 H, COCI-C 6 -alkyl, CO2C1-C 6 -alkyl, CONR 1 aR2, NRiaR2, NR 1 aCOC 1

C

6 -alkyl and any two of adjacent substituents can be joined to form a 6or 7- membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, heteroaryl, wherein heteroaryl is defined as a or 6-membered ring substituted with one and two heteroatoms selected from O, S, N, unsubstituted or substituted with one, two or three [N:\LIBAA]O1587.doc:TAB substituents selected from the group consisting of: Br, Cl, F, 1, (01-06)alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONRlaR2, NRlaR 2 NRIaCOCI -C 6 -alkyl, any two adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring: Rla and R 2 are independently: a) (Cl-C 6 )-alkyl, or b) aryl, wherein aryl is phenyl or naphthyl; o 3 is: a) -(C 1

-C

6 )-alkyl, alkyl as defined above; b) -(C 1

-C

6 )-alkenyl, wherein alkenyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COCI-C 6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONRlaR 2 NRlaR 2 NRlaCOC 1

-C

6 -alkyl, aryl as defined above, and heteroaryl as defined above; C) -(Cl-C 6 )-alkynyl, wherein alkynyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COCI-C 6 -alkyl, C0201 -C 6 -alkyl, CONR 1 aR 2 NRU R2, NR 1 aCOCi -C 6 -alkyl, aryl as defined 20 above, and heteroaryl as defined above, d) -aryl, aryl as defined above, or e) -heteroaryl, heteroaryl as defined above;

R

4 is: a) absent and a is a double bond:

S

egos

C

S S

S.

5655 5 0*

C

5500 0C

S.

OSS

S C 0e 0 *000 0 0 0* 9

S.

0 0

S

25 b) -H, c) -OH, d) =0, e) -O(=)rs1C0akl alkyl as defined above, f) -O[(C=O)0r]sC2-Clo-alkenyl, as defined above, g) -O[(C=O)O]sC 2

-C

6 -alkynyl, alkynyl as defined above, h) -O[(C=O)Orl(C3-C 7 )-cycloalkyl, i) -O[(C=O)Or]saiyl, aryl as defined above, j) -O[(C=O)Or]sheteroaryl, heteroaryl as defined above, k) -O(CH 2 )nO(CH 2 )6heteroaryl, heteroaryl as defined above, 1) -O(CH2)nO(CH 2 )maryl, aryl as defined above, m) -OC(=O)NRlaR2, n) -0S0 2

R

3 or o) -NRlaR 2

R

2 9a and R29b are independently: a) -H, b) -(CH 2 )s-O[(C=O)Or]sC 1 -Ci-alkyl, alkyl as defined above, (N:\LIBAA]O1 587.doc:TAB

M

c) -(CH 2 )s-0[(C=0)Or]sC 2 -Clo-alkenyl, alkenyl as defined above, d) -(CH 2 )s-0[(C0)Or]sC 2

-C

6 -alkynyl, alkynyl as defined above, e) -(CH 2 )S-0[(C=0)Orls(C 3

-C

7 )-cycloalkyl, f) (CH2)s-O[(C=O)Or]saryl, aryl as defined above, g) -(CH 2 )s-0[(C=0)Orlsheteroay, heteroaryl as defined above, h) -(CH 2 )s-0(0H 2 )nO(CH 2 )mheteroaryl, heteroaryl as defined above, i) -(CH 2 )s-0(CH 2 )flO(0H 2 )maryl, aryl as defined above, j) -(CH 2 )s-0C(=0)NRlaR 2 k) -(CH 2 )s-0S0 2

R

3 1) -(Cl-C 6 )-alkyl, alkyl as defined above, or m) -(0 2 -0 6 )-alkenyl, alkenyl as defined above; R29a and R29b together are =0 or =C(Cl-0 10 -alkyl) 2 alkyl as defined above.

An embodiment of this embodiment of the invention are the compounds of Formula I R 2a R 2b 1 H1819 2 0 c 1 H 1 8 1 7 22 0 rI1 H q I-I28 C0 2

CH

3 0O S S S

OS

S

5* 59 0*@S

S

S..

'0Ac or a pharmaceutically acceptable salt, crystal form, or hydrate, wherein: *X is: 0; a is: a single bond; b band care 0 independently: a single bond or a double bond; 0*2on is: i to 4; m is: 1 to 4; r is: 0Corn; s sis: 0Corn;

R

1 is: a) H, or b) (01-06)-alkyl, wherein alkyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, vinyl, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, C0C 1 -0 6 alkyl, 00201 -C 6 -alkyl, C0NR 1 aR 2 N R 1 aR2, NRP 1 aCoG 1

-C

6 -alkyl, aryl, wherein aryl is defined as phenyl or naphthyl, unsubstituted or substituted F(Al./with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO, [N:\LIBAA]O1 587.doc:TAB

CO

2 H, COC1 -C 6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONRlaR2, NR 1 aR2, NR 1 aCOC 1

C

6 -alkyl and any two of adjacent substituents can be joined to form a 6or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, heteroaryl, wherein heteroaryl is defined as a 5 or 6-membered ring substituted with one and two heteroatoms selected from 0, S, N, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONRlaR2, NRlaR 2 NRlaCOC 1

-C

6 -alkyl, any two adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring: Rla and R 2 are independently: a) (C 1

-C

6 )-alkyl, or b) aryl, wherein aryl is phenyl or naphthyl,

R

3 is: a) -(Cl-C 6 )-alkyl, alkyl as defined above, b) -ayay*sdeie bvo b) -hraryl, traryl as defined above,

R

4 is: a) -O[(C=O)OJsC 1

-C

10 -alkyl, alkyl as defined above, b) 0 [(C=)Ors(3-C7)cycloalkyl, c) -O[(C=O)Or]saryl, aryl as defined above, d) -O[(C=O)O]sheteroaryl, heteroaryl as defined above, e) -O(CH2)nO(CH 2 )mheteroaryl, heteroaryl as defined above, -O(CH2)flO(CH2)marjyl, aryl as defined above, o.*25 g) -OC(=O)NRlaR2, or h) -0S0 2

R

3

R

2 9a and R29b are independently: a) -H, b) -(CH2)s-O[(C=0)O]sC 1

-C

1 0 -alkyl, alkyl as defined above, c) -(CH2)s-O[(C=O)0]sC 2

-C

1 Q-alkenyl, alkenyl as defined above, d) -(CH2)s-0[(C=O)]sC 2

-C

6 -alkynyl, alkynyl as defined above, Se) -(CH2)s-O[(C=0)OrIs(C 3

-C

7 )-cycloalkyl, f) -(CH2)s-O(C=O)0]saryl, aryl as defined above, g) -(CH2)s-O[(C0)O]sheteroaryl, heteroaryl as defined above, h) -(CH2)s-O(CH2)flO(CH 2 )mheteroaryl, heteroaryl as defined above, i) -(CH2)s0(CH 2 )fl0(CH 2 )maryl, aryl as defined above, j) -(CH 2 )s-0C(=O)NRlaR 2 k) -(CH 2

),-OSO

2

R

3 1) -(C 1

-C

6 )-alkyl, alkyl as defined above, or M) -(C 2

-C

6 )-alkenyl, alkenyl as defined above; [N:\LIBAA]O1 587.doc:TAB 12 R29a and R29b together are =0 or =C(ClCl-alkyl)2, alkyl being as defined above.

An embodiment of this embodiment are the compounds of structural Formula I or a pharmaceutically acceptable salt, crystal form or hydrate, wherein: R4 is: a) -0[(C=O)Orlsaryl, wherein aryl is defined as phenyl or naphthyl, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C)-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO, C02H, COC-C-alkyl, C02C1-6-alkyl, CONRiaR2, NRlaaR2, NRlaCOC -C-alkyl and any two of adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or b) -0[(C=O)Orlsheteroaryl, wherein heteroaryl is defined as a 5 or 6-membered ring substituted with one and two heteroatoms selected from 0, S, N, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C6)-alkoxy, cyano, nitro, hydroxy, CHO, C02H, COC1-C-alkyl, C02C1-C6-alkyl, CONRiaR2, NRiaR2, NRlaCOC -C-alkyl, any two adjacent substituents can be joined .i to form a 6- [N:\LIBAA01 587.doc:TAB 13 THIS PAGE HAS BEEN LEFT INTENTIONALLY BLANK

S

S. S S S

S.

OS@O

S

*0

S

OS**

0* 0*S 0O 0

S

0@ 0 00 0 0

S

[N:\LIBAAIO 1 587.doc:TAB 14 THIS PAGE HAS BEEN LEFT INTENTIONALLY BLANK 0Oee *00 609 0S [N:\LIBAA]O1 587.doc:TAB THIS PAGE HAS BEEN LEFT INTENTIONALLY

BLANK

4 4 *4eO *4 0 0~ 00 404*

S

OS

0 0006 00

OS

aS.

555.

0e S. 55** 55 55 0 @0 S. [N:\LIBAAJO1 587.doc:TAB WO 97/16182 PCT/US96/17482 16or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring.

An embodiment of the invention are the compounds of Formula I R29b C 12 OAc 3 OAc 23 a R 4 or a pharmaceutically acceptable salt, crystal form, or hydrate, wherein: X is: H and R1; a is: a single bond; b and c are independently: a single bond or a double bond; n is: m is: r is: s is: 1 to 4; 1 to 4; Sor 1; 0or 1;

R

1 is: a) H, or b) (C 1

-C

6 )-alkyl, wherein alkyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I,

(C

1

-C

6 )-alkoxy, vinyl, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC1-C6alkyl, C0201-C 6 -alkyl, CONR 1 aR2, NR 1 aR2, NR 1 aCOC 1

-C

6 )-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO,

CO

2 H, COC 1

-C

6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONR 1 aR 2

NR

1 aR2, NR 1 aCOC 1

C

6 -alkyl and any two of adjacent substituents can be joined to form a 6or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, heteroaryl, wherein heteroaryl is defined as a 5 or 6-membered ring substituted with one and two heteroatoms selected from O, S, N, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1

-C

6 )-alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONRIaR2, NRlaR 2 NRlaCOC-C-alkyl, any two adjacent substituents ca be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen *atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring;

R

1 a and R 2 are independently: S*a)

(C

1

-C

6 )-alkyl, or b) aryl, wherein aryl is phenyl or naphthyl;

R

3 is: a) -(C 1

-C

6 )-alkyl, alkyl being as defined above; b) -(C 1

-C

6 )-alkenyl, wherein alkenyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, I, *o :(C 1

-C

6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONRIaR 2 NRlaR 2 NRlaCOC 1 -C6-alkyl, aryl as defined above, and heteroaryl as defined above; c) -(C 1

-C

6 )-alkynyl, wherein alkynyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I,

*(C

1

-C

6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 -alkyl, C0 2 0 1

-C

6 -alkyl, CONRlaR 2 NRlaR 2 NRlaCOC 1 -C6-alkyl, aryl as defined above, and heteroaryl as defined above, d) -aryl, aryl as defined above, or e) -heteroaryl, heteroaryl as defined above;

R

4 is: a) absent and a is a double bond; b) -H, BAL' c) -OH, 4' d) =O, IN:\LIBAA]O1587.doc:TAB 18 e) O0[( 0 0)Or~sCi -CjO -alkyl, alkyl as defined above, f) 0 [I(C=O)OrIsC 2 -Cio-alkenyl, as defined above, g) -O[(C=O)Or~sC 2

-C

6 -alkynyl, alkynyl as defined above, h) 0 [I(C=O)Or~s(0 3

-C

7 )-cycloalky, i) O0[(C=O)Orlsaryl, aryl as defined above, j) -O[(C=O)Orlsheteroaryl, heteroaryl as defined above, k) -O(CH2)nO(0H 2 )mheteroaryl, heteroaryl as defined above, 1) -O(CH2)flO(CH 2 )maryl, aryl as defined above, m) -OC(=O)NRlaR 2 n) -0S0 2

R

3 or o) -NRlaR 2 R29a and R29b are independently: a) -H, b) -(CH2)sOf(C0)Or~sC-ClO-alkyl, alkyl as defined above, c) -(CH2)s-O[(C=O)Or]sC 2 -Clo-alkenyl, alkenyl as defined above, d) -(CH2)s-O[(C=O)0r]sC 2

-C

6 -alkynyl, alkynyl as defined above, ae) -(CH2)s-O[(C=O)Orls(C 3

-C

7 )-cycloalkyl, 4, f) -(CH2)s-O[(C=O)Or]saryl, aryl as defined above, g) C 2 s C r et r a y e e o r l s d f n d a o e h) -(CH2)s0O(C 2

)O(CH

2 heteroaryl heteroaryl as defined above, 6QOBi) -(CH2)s 0

(H)O(CH

2 )maryl, aryl as defined above, j) -(CH 2 )s-0C(=O)NRlaR 2 k) -(CH 2 )s-OS0 2

R

3 1) -(C 1

-C

6 )-alkyl, alkyl as defined above, or M) -(C 2

-C

6 )-alkenyl, alkenyl as defined above; a R29a and R29b together are =0 or =C(0 1

-C

10 -alkyl) 2 alkyl being as defined above.

400*:* aAn embodiment of this embodiment of the invention are the compounds of Formula 1: R29a 29b 19 2 c 12 1j 8 1 7 22 0 2 1H H OH 2800 2

CH

3 H9 I 14 16 3 10O25 2 5 QAc 0 24 5 6 7. QAc OAc 4 OAc 2Ta or a pharmaceutically acceptable salt, crystal form, or hydrate, wherein: 3, X is: H and R 1 is a single bond; IN:\LIBAA]O1 587.doc:TAB b and c are independently: n is: m is: ris: s is: a single bond or a double bond; 1 to 4; 1 to 4; 0 or 1; 0 or 1; 0000 00000 000 *0 *6 so fe"00090 *00 060 a 0 0 a* 0 00.

00 0 0 o 0 05 000 5500 0@ S 00 *0 0 000 000 0 0 00 Se 0 55 000 O 0 *00 @500 0 000.

0 O 0 R1 is: a) H, or b) (C 1

-C

(C

1

-C

6 )-alkoxy, vinyl, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 alkyl, C0 2

C

1

-C

6 -alkyl, CONRIaR2, NR 1 aR2, NR 1 aCOC 1

-C

6 )-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO,

CO

2 H, COC1-C 6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONR 1 aR2, NR 1 aR2, NR 1 aCOC 1

C

6 -alkyl and any two of adjacent substituents can be joined to form a 6or 7- membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, heteroaryl, wherein heteroaryl is defined as a 5 or 6-membered ring substituted with one and two heteroatoms selected 20 from O, S, N, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I,

(C

1

-C

6 )-alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 -alkyl, C0 2

C

1

C

6 -alkyl, CONRlaR 2 NRlaR 2 NRlaCOC 1 -C6-alkyl, any two adjacent substituents can be joined to form a 6- or 7-membered fused ring said 25 ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring: Rla and R 2 are independently: a) (C 1

-C

6 )-alkyl, or b) aryl, wherein aryl is phenyl or naphthyl; R3 is: a) -(C 1

-C

6 )-alkyl, alkyl as defined above; b) -(C 1

-C

6 )-alkenyl, wherein alkenyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I,

(C

1

-C

6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 -alkyl, C0 2

C

1

-C

6 -alkyl, CONRlaR 2 NRIaR2, NRlaCOC 1

-C

6 -alkyl, aryl as defined above, and heteroaryl as defined above; c) -(C 1

-C

6 )-alkynyl, wherein alkynyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, I,

(C

1

-C

6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1

-C

6 -alkyl, [NALIBAAO1 587.doc:TAB

R

4 is: C0 2

C

1

-C

6 -alkyl, CONRlaR2, NRlaR 2 NRlaCOCI-C6-alkyl, aryl as defined above, and heteroaryl as defined above, d) -aryl, aryl as defined above, or e) -heteroaryl, heteroaryl as defined above; a) -OH, b) O0[(C=O)OrsC-CiO-alkyl, alkyl as defined above, C) -O[(C=O)Or~s(C 3 0C 7 )-cycloalkyl, d) -O[(C=O)Or]saryl, aryi as defined above, e) -O[(C=O)Or4sheteroaryl, heteroaryl as defined above, f) -O(CH-2)nOCH- 2 )mheteroaryl, heteroaryl as defined above, g) -O(CH 2 )nO(0H 2 )maryl, aryl as defined above, h) -OC(=O)NRIaR 2 or i) -0S0 2

R

3 independently:

R

2 9a and R29b are a) -H, b) -(CH2)s-O[(C=O)OrsC-ClO-alkyl, alkyl as defined above, c) -(CH2)s-O[(C=O)Or]sC 2 -ClO-akeny, alkenyl as defined above, d) -(CH 2 )s-O[(C=O)Or]sC 2

-C

6 -alkynyl, alkynyl as defined above, 2*0690 e) -(CH2)s0[I(C=O)Orls(C 3

-C

7 )-cycloalkyl, 0006a 2 (CH2)s-O[(C=O)Orsary, aryl as defined above, -(CH2)s-O[(C=O)Or]sheteroaryl, heteroaryl as defined above, see..h) -(CH2)s0(CH2)nO(CH 2 )mheteroaryl, heteroaryl as defined above, S..*soi) -(CH2)s0(CH 2 )nO(CH 2 )maryl, aryl as defined above,

-(CH

2 )s-OC(=O)NRlaR 2 25 k) -(CH 2 )s-OSO 2

R

3 1) -(0 1

-C

6 )-alkyl, alkyl as defined above, or M) -(0 2

-C

6 )-alkenyl, alkenyl as defined above;

R

2 9a and R29b together are =0 or =C(Cl-0 10 -alkyl) 2 alkyl being as defined above.

An embodiment of the invention is a compound selected from the group consisting of: 4,6,7,15,1 6-pentakis(acetyloxy)-21 ,22-epoxy-1 8-hydroxy-22-methoxycarbonyl[60c,7a, 1 5P3,1 6j3, 20a,21 l j 22 f 3 ID:A-Friedo-A-homo-27,30.dinor24oxaoleanal1 -en-3-one; 4,6,7,15,1 6-pentakis(acetyloxy).21 ,22-epoxy-1 8-hydroxy-22-methoxycarbonyl[6c7a, 1 503,16p3, 2oc,21 I3,22 1 3 D:A-Friedo-A-homo-27, 3 0-dinor-24-oxaoleanalI -ene; 4,6,7,15,1 6-pentakis(acetyloxy)20,21 ,22,(29)-diepoxy-1 8 -hydroxy-22-methoxycarbonyl[6cL,7cc 15P3, 16f3,2Oc,21 j3,22f3]D:A-Friedo-A-homo-27,30-dinor-24-oxaoleana-1 -en-3-one; (N:\LIBAAIO1 587.doc:TAB THIS PAGE HAS BEEN LEFT INTENTIONALLY BLANK e.g.

000@@ S e,@.e cc..

Ce C CC C C C C Ce ge e.

eec.

eeeee C Ce~eg eec.

C Cc e gee c c C C Ceeec CeeC se es C e e e Cc.

e eec

*CCC

eec~ e e e e *e c se C ecee e e e ce Ce C e eq cc C C *ecc eec.

[N:\LIBAA]O1 587.doc:TAB THIS PAGE HAS BEEN LEFT INTENTIONALLY BLANK

S

S@S**

S'S.

55 @5 5 6 5 @0 @5 55 0555 *5660 5500@ .55.

5* 0@5 S S S *SS*s 5055 55 5 5 S S See

*SS

*05@ 0*Se 0 S 55 55 5 SOS. S S 5 55 0 55 5 55 55 S S S S *5*S

OSSO

055 S S 555550 5 [N:\LIBAA]O1 587.doc:TAB WO 97/16182 WO 9716182PCT/US96/17482 23 4,6,7,15,1 6-pentakis (acetyloxy)-2 1,22-epoxy- I 8-hydroxy-22methoxycarbonyl [6ux,7, I 5p, 1 6P,21 f,2213]D:A-Friedo-A-homo-27,30.

dinor-24-oxaoleana- 1 -en-3 29(Z)-methoxycarbonyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- 1 8-hydroxy-22-methoxycarbonyl [6x,7a, 1513,1613,2113,223] D:A-Friedo- A-homo-27,30-dinor-24-oxaoleana- 1 ,20(29)-dien-3-one; 29(E)-methoxycarbonyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- 1 8-hydroxy-22-methoxycarbonyl[6cx,7ux, 1$0,1613,210f,221]D:A-Friedo- A-homo-27 3 O-dinor-24-oxaoleana- I ,20(29)-dien-3 -one; 29(Z)-t-butoxycarbonyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- 1 8 -hydroxy-22-methoxycarbonyl [6a,7Q, 15Sf31 61,210f,22f]D:A-Friedo- A-homo-27,30-dinor-24-oxaoleana- 1 ,20(29)-dien-3-one; 29(E)-t-butoxycarbonyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- 1 -hydroxy-22-methoxycarbonyl r6(x,7ax, 15p3, 1 6,21 1,22f3]D:A-Friedo- A-homo-27 3 O-dinor-24-oxaoleana- 1 ,20(29)-dien-3-one; 29(Z)-ethoxycarbonyl-4,6,7,15, 16-pentakis(acetyloxy)-2 1,22-epoxy-i 18hydroxy-22-methoxycarbonyl[6Qx,7a, 1513,1613,21 1,221]D:A-Friedo-Ahomo-27 3 0-dinor-24-oxaoleana- 1 ,20(29)-dien-3-one; 29(E)-ethoxycarbonyl-4,6 ,7,15,1 6-pentakis(acetyloxy)-2 1,22-epoxy-I 18hydroxy-22-methoxycarbonyl[6a,7Qx 1 513,1613,21 1,221]D:A-Friedo-Ahomo-27 3 0-dinor-24-oxaoleana- 1 ,20(29)-dien-3-one; 29(Z)-cyano-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- I 8-hydroxy- 22-methoxycarbony1[6ux,7c, 1513,1613,2101,2213D:A-Friedo-A-homo- 2 7,30-dinor-24-oxaoleana- I ,20(29)-dien-3-one; WO 97/16182 WO 9716182PCTIUS96/1 7482 24 29(E)-cyano-4,6,7, 15,1 6-pentakis(acetyloxy)-2 l, 2 2-epoxy-1I 8-hydroxy- 22-methoxycarbonyl[6a,7cc, 1513,1613,21 1,22f3]D :A-Friedo-A-homo- 27,30-dinor-24-oxaoleana- 1 ,20(29)-dien-3 -one; 29(E)-phenyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1, 2 2-epoxy- I 8-hydroxy- 22-methoxycarbonyl[6ac,7Q, 1513,1613,21 1,2213]D :A-Friedo-A-homo- 27,30-dinor-24-oxaoleana- I ,20(29)-dien-3 -one; 29(Z)-phenyl-4,6,7,15,1 6 -pentakis(acetyloxy)-2 l,22-epoxy- I 8-hydroxy- 22-methoxycarbonyl [6Qx,7u, 1513,1613,2113,221]D :A -Friedo-A-homo- 2 7,3 0-dinor-24 -oxao lean a- 1 ,20(29)-dien-3 -one; 4-(2-bromobenzoyl)oxy-6 ,7,15,1 6-tetrakis (acetyloxy)-2 1,22-epoxy-i 8hydroxy-22-methoxycarbonyl [6cx,7, 1 51,1 61,20a,2 101,22f]D:A- Friedo-A-homo-27 3 O-dinor-24-oxaoleana- I -en-3 -one; 4,6,7,15,16 6-pentaki s(acety lox y) -21,22-epox y- I 8-hydroxy-22-methoxycarbonyl [6oc,7a, 1 5p3, 1 6,20oc,21 1,221]D:A-Friedo-A-homo-27,30.

dinor-24 -oxaoleana-3 -one; 4,6,7,15,1 6-pentakis (acetyloxy)-2 1,22-epoxy- I 8-hydroxy-22methoxycarbonyl[6cc,7uc, 1 513,1 61,20c,2 1 1,221]D:A-Friedo-A-homo- 2 7,30-dinor-24-oxaoleane; 4,6,7,15,1 6-pentakis(acetyloxy)-20(29),2 1,22-diepoxy- 1 8-hydroxy-22methoxycarbonyl[6u,7(x, 1 51,1 61,20a,2 1 1,221]D:A-Friedo-A-homo- 2 7 3 O0-dinor-24-oxaoleana-en-3 -one; 4,6,7,15,1 6-pentakis (acetyloxy)-2 1,22-epoxy-i I8-hydroxy-22methoxycarbonyl [6(x,7x, 1513,1 6 1,20x,2 113,223]D :A-Friedo-A-homo- 2 7 ,30-dinor-24-oxaoleana-3 WO 97/16182PCIS6178 PCT/US96/17482 25 29(Z) -methox ycarbonyl -4,6,7,15,1 6-pentaki s(acety loxy) -21l, 2 2 -epoxy- 1 8 -hydroxy-22-methoxycarbonyl [6ix,7u, 15p3,1 61,2101,221]D:A-Friedo- A -hono 2 7,3O-dinor-24-oxaoleana-2O(29)en-3 -one; 29(E)-methoxycarbonyl-4,6 15,1 6-pentakis(acetyloxy)-2

I,

2 2-epoxy- 1 8-hydroxy-22-methoxycarbonyl [6(x,7ix, 1503,1603,2 1 1,221]D :A-Friedo- A -homo-27 3 O-dinor- 2 4-oxaoleana-20(29)en-3.-one; 29(Z)-t-butoxycarbonyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1, 2 2-epoxy- I8-hydroxy-22-methoxycarbonyl[6x,7Qx, 15p3,1613,2101,221]D:A-Friedo- A-homo-27 3 O-dinor- 2 4-oxaoleana-2O(29)-en-3.-one; 29(E)-t-butoxycarbonyl-4,6,7,15,1I6-pentakis(acetyloxy)-2 1,22-epoxy- 1 8-hydroxy-22-methoxycarbonyl [6u,7Qx, 1513,1613,2101,22131DA-Friedo- A-homo-27 30 -dinor-24-oxaoleana-20(29y..en-3 -one; 29(Z)-ethoxycarbonyl-4,6,7,ls,1 6-pentakis(acetyloxy)-2 1,22-epoxy-i 18hydroxy-22-methoxycarbonyl [6Qx,7Q, 1513,1613,2113,2213] DA-Friedo-Ahomo- 2 7 3 O-dinor-24-oxaoleana2(29)-en-3 -one; 29(E)-ethoxycarbonyl-4,6 ,7,15,1 6-pentakis(acetyloxy)-2 l.22-epoxy- 18hydroxy-22-methoxycarbonyl[6a,7Q, 1513,1613,21 j3,221]D:A-Friedo-Ahomo-27 3 O-dinor-24-oxaoleana-20(29)-en-.3 -one; 29(Z)-cyano-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- I 8-hydroxy- 2 2-methoxycarbonyl[6&x,7a, 1513,1613,21 1,221DA-Friedo-A-homno- 2 7 3 O-dinor- 2 4 -o xaoleana-20(29) en 3.-one; 29(E) -cyano-4,6,7,15,1 6 -pentaki s(acetylIoxy)-2 1,2 2-epoxy- I 8-hydroxy- 2 2 -methoxycarbony1[6a,7Qx, 1513,1613,21 1,2213D:A-Friedo-A-homo- 2 7 3 O0-dinor- 2 4 -oxaoleana-20(29)en-3.-one; 29(E)-phenyl-4,6,7,15,1 6 -pentakis(acetyloxy)-2 l, 22 -epoxy-1I 8-hydroxy- 22-methoxycarbonyl [6(x,7ux, 1 5p3,1613,21 1,2213]D:A-Friedo-A-homno- 2 7 3 O-dinor-24-oxaoleana-20(29)-en-.3-one; WO 97/16182 PCT/US96/17482 -26- 29(Z)-phenyl-4,6,7,15,16-pentakis(acetyloxy)-21,22-epoxy-18-hydroxy- 22-methoxycarbonyl[6a,7a, 150,16p,2103,22]D:A-Friedo-A-homo- 2 7,30-dinor-24-oxaoleana-20(29)-en-3-one; or 4-(2-bromobenzoyl)oxy-6,7,15,16-tetrakis(acetyloxy)-21,22-epoxy-18hydroxy-22-methoxycarbonyl[6a,7a, 15p, 16p,21 0,22p]D:A-Friedo-Ahomo-27,30-dinor-24-oxaolean-3-one.

The compounds of the present invention have asymmetric centers and this invention includes all of the optical isomers and mixtures thereof.

In addition compounds with carbon-carbon double bonds may occur in Z- and E- forms with all isomeric forms of the compounds being included in the present invention.

As used herein, the term "alkyl" includes those alkyl groups of a designated number of carbon atoms of either a straight, branched, or cyclic configuration. Examples of "alkyl" include methyl, ethyl, propyl, isopropyl, butyl, sec-and tert-butyl, pentyl, hexyl, heptyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbomyl, and the like. "Alkoxy" represents an alkyl group of indicated number of carbon atoms attached through an oxygen bridge, such as methoxy, ethoxy, propoxy, butoxy and pentoxy.

"Alkenyl" is intended to include hydrocarbon chains of a specified number of carbon atoms of either a straight- or branchedconfiguration and at least one unsaturation, which may occur at any point along the chain, such as ethenyl, propenyl, butenyl, pentenyl, dimethyl pentenyl, and the like, and includes E and Z forms, where applicable. "Halogen", as used herein, means fluoro, chloro, bromo and iodo.

The term "aryl" is defined as a phenyl or naphthyl ring which is optionally substituted at any available carbon atoms with one, two or three substituents selected from the group consisting of: Br, Cl, F, I, (C1-C6)-alkoxy, phenyl, phenoxy, cyano, oxo, nitro, hydroxy, WO 97/16182 PCT/US96/1 7482 -27- CHO, CO2H, COCl-C6-alkyl, CO2Cl-C6-alkyl,

CONR

1

R

2

NR

1

R

2 NRI COC -C6-alkyl. The aryl may also be substituted with a fused or 7-membered ring containing one or two oxygens and the remaining ring atoms being carbon, the fused or 7-ring being selected from the group consisting of: dioxolanyl, dihydrofuranyl, dihydropyranyl, and dioxanyl.

The term "heteroaryl" as utilized herein is intended to include the following a 5 or 6-membered ring substituted with one or two heteroatoms selected from O, S, N, and is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, I, (Cl-C6)-alkoxy, cyano, nitro, hydroxy, CHO, CO2H, COC -C6-alkyl, CO2C1-C6-alkyl, CONR1R 2

NR

1

R

2 NR ICOC -C6-alkyl, any two adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring. Heteroaryl groups within the scope of this definition include but are not limited to: acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrrazolyl, indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, and pyrrolyl which are substituted or unsubstituted as defined above.

In the compounds of Formula I, the heteroaryl group may be optionally substituted with the substituents listed above at any available carbon atom or nitrogen atom (if present), but compounds bearing certain substitutents, directly substituted to a nitrogen may be relatively unstable and are not preferred. The heteroaryl may also be fused to a second or 7-membered ring containing one or two oxygens selected from the the remaining ring atoms being carbon, selected from the group consisting of: dioxolanyl, dihydrofuranyl, dihydropyranyl, and dioxanyl.

Pharmaceutically acceptable salts include both the metallic (inorganic) salts and organic salts; a list of which is given in Remington's Pharmaceutical Sciences, 17th Edition, pg. 1418 (1985). It is well known to one skilled in the art that an appropriate salt form is WO 97/16182 PCT/US96/17482 -28chosen based on physical and chemical stability, flowability, hydroscopicity and solubility. As will be understood by those skilled in the art, pharmaceutically acceptable salts include, but are not limited to salts of inorganic acids such as hydrochloride, sulfate, phosphate, diphosphate, hydrobromide, and nitrate or salts of an organic acid such as malate, maleate, fumarate, tartrate, succinate, citrate, acetate, lactate, methanesulfonate, p-toluenesulfonate or palmoate, salicylate and stearate. Similarly pharmaceutically acceptable cations include, but are not limited to sodium, potassium, calcium, aluminum, lithium and ammonium (especially ammonium salts with secondary amines).

Preferred salts of this invention for the reasons cited above include potassium, sodium, calcium and ammonium salts. Also included within the scope of this invention are crystal forms, hydrates and solvates of the compounds of Formula I.

WO 97/16182 PCT/US96/17482 -29- REACTION SCHEME A 'OAc Li, NH 3

THF

OAc As seen in Scheme A, compound I, 4 ,5,6,15,16-pentakis (acetyloxy)-21,22-epoxy-1 8 -hydroxy-22-methoxycarbonyl[6X,7cx, 153,16p,21 P, 2 2 3 ]D:A-Freido-A-homo-27,30-dinor-24-oxaoleana- 1,20(29)-diene-3-one, isolated from Spachea correa in liquid ammonia with lithium metal will result in the reduction of the Cl olefin group to produce the saturated lactone. Alternative methods for reducing the Cl olefin group and/or the C20(29) olefin that are known in the art may also be employed. US Serial Number 08/476,806 filed on June 7, 1995 describes the isolation of compound I and is hereby incorporated by WO 97/16182 PCT/US96/17482 reference. The resultant lactone can then be converted to the oxepin analog by procedures described in Reaction Scheme B.

It should also be noted that compounds of Formula I having the 11,12-double bond can be prepared using the starting material, 4,6,7,15,16-pentakisacetoxy-21,22-epoxy- 8-hydroxy-22methoxycarbonyl [6a,7a,15, 16p,21P,22]D :A-Freido-A-homo-27,30dinor-24-oxaleana-1,1 1,20(29)-trien-3-one, isolated from Spachea correa and following the procedures described herein. However, there may be reactions where it will not be possible to selectively operate on one of the double bonds, for example, ozonolysis.

WO 97/16182 PCT/US96/17482 -31 REACTION SCHEME B OAc I (b double bond) II (b single bond) 1. LiAIH(OtBu) 3 2. Et 3 SiH, BF 3 OEt 2 'OAc As seen in Scheme B, compound I [4,6,7,15,16pentakis(acetyloxy)-21,22-epoxy-18-hydroxy-22-methyoxycarbonyl- [6a,7a, 15p, 160,21 p,22p]D:A-Freido-A-homo-27,30-dinor-24oxaoleana-1, 20(29)-dien-3-one], isolated from Spachea correa can be converted to its oxepin analog in a two step process. US Serial Number 08/476,806 filed on June 7, 1995 describing the isolation of compound I and is hereby incorporated by reference. Lactone I is first reduced to the lactol. This can be accomplished by using a variety of reducing agents including di-isobutylaluminum hydride (Dibal) and sodium bis(2methoxyethoxy)aluminum hydride (Red-Al). A more optimal reducing WO 97/16182 PCT/US96/17482 -32agent is the use of lithium tris-t-butoxyaluminum hydride in an inert solvent such as dichloromethane at reduced temperatures, preferably 0°C. The purified lactol intermediate is then reacted with triethylsilane and a Lewis acid such as borontrifluoride diethyl etherate to give the ether (oxepin) analog of I.

REACTION SCHEME C OAc 1. LiAIH(OtBu) 3 2. Et 3

AI

'OAc I"OAc OAc In a variation of Scheme B, oxepin derivatives substituted at C3 can also be prepared. Thus in Reaction Scheme C, lactone I or its reduced analog is first reduced to the lactol as described in Reaction Scheme B. The purified lactol intermediate is then reacted with a WO 97/16182 PCT/US96/17482 -33 trialkylaluminum reagent, as exemplified in this scheme by triethylaluminum (Et3AI) to give the ethyl derivative. The allyl derivative can be prepared with allyltrimethylsilane and a Lewis acid such as borontrifluoride diethyl etherate.

REACTION SCHEME D

'CO

2

CH

3

HCI/THF

or

CH

3

(CI)AI[N(OCH

3 )C H 3 Lactone or ether derivatives can be selectively deacetylated at C4 to give the corresponding alcohol by reacting it withan aqueous solution of HCI (preferably 2M to 3M concentration) in THF.

It can also be prepared by reacting I with CH3(C1)Al[N(OCH 3

)CH

3 WO 97/16182 PCT/US96/17482 -34- (Weinreb reagent) in inert solvents such as THF, toluene or methylene chloride.

REACTION SCHEME E Ac CH3 4 The C4 hydroxy group can be oxidized to the corresponding ketone by a variety of oxidizing agents. The Jones reagent (chromic acid and sulfuric acid in H20), pyridinium chlorochromate, and oxalyl chloride plus DMSO all will achieve this conversion.

WO 97/16182 PCT/US96/17482 REACTION SCHEME F 'OAc (PhO) 3 MePI HMPT, 75 °C j PO)MP .0 C0 2

H

3 'OAc The C4 hydroxy group can also be dehydrated to give the olefin. Reaction of the alcohol with tris-phenoxymethylphosphonium iodide in hexamethylphosphorous triamide (HMPT) at 75 0 C will achieve this conversion.

WO 97/16182 WO 9716182PCTIUJS96/17482 36 REACTION SCHEME G R 4 'COCI M 0 H~ O Oc 'O2C

H

Me Me Oc

Q~MA

OO~c Me~~M 0 1R4 1 D! O 1 2 H U Li 0 2

C

NR

1 R 2 WO 97/16182 PCT/US96/17482 37- REACTION SCHEME G (CONT'D) CDI, R4'OH

CO

2

CH

3

R

4 'OH, Tf20, base 'OAc O- OAc UAc

CH

3 4 OR4' As depicted in Reaction Scheme G, esters at C4 can be prepared by reaction of a preformed carboxylic acid chloride with the C4 alcohol derivative (Reaction Scheme D) in a basic solvent such as pyridine. It should be understood that R 4 is used to represent a portion of the R 4 definition, e.g. R 4 can be an alkyl carbonate which is depicted in the scheme as OC(=0)OR 4

R

4 representing the alkyl substituent.

The acid chlorides, when not purchased, are prepared by stirring the carboxylic acids in reagents such as oxalyl chloride or thionyl chloride.

Esters may also be prepared by reaction of the acid chloride and C4 alcohol with silver cyanide (AgCN) in an aprotic solvent such as HMPA.

WO 97/16182 PCT/US96/17482 -38 C4 sulfonate derivatives are prepared in a similar manner by reaction with sulfonyl chlorides.

C4 carbonate and carbamate derivatives are prepared by first reacting the C4 alcohol derivative with carbonyldiimidazole (CDI) to obtain the imidazolecarbonyl intermediate which is then reacted with an alcohol or amine (R 1

R

2 NH) to give the corresponding carbonate or carbamate derivatives.

C4 ether derivatives can also be prepared. The best procedure involves reacting an alcohol with trifluoromethanesulfonic anhydride (Tf20, triflic anhydride) to obtain the preformed triflate in dichloromethane at reduced temperature, preferably -78 0 C. To this solution is added the triterpene alcohol, the reaction mixture is warmed to room temperature and stirring is continued until reaction is complete.

Ethers may also be prepared by heating a mixture of triterpene C4 alcohol, the appropriate alkylhalide and an excess of silver oxide in an aprotic invert solvent such as THF.

WO 97/16182 PCT/US96/17482 -39- REACTION SCHEME H C0 2

CH

3

CH

3

NHR

1

R

2 NaCNBH 3

H'R

C0 2 0H 3 Amines at C4 can be prepared from the C4 ketone described in Reaction Scheme E by reaction with an amine NHR 1

R

2 in a variety of solvents with a reducing agent such as sodium cyanoborohydride.

WO 97/16182 PCT/US96/17482 REACTION SCHEME I 'OAc OAc [Cs 6

H)

3

P]

3 RhCI THF, 80 psi OAc 24 _6A OAC OAc 21 -R 4 Lactone or ether (oxepin) derivatives which have or have not been derivatized at C4 can be selectively hydrogenated at the position to give the methyl analog. This conversion can be achieved by hydrogenation with Tris(triphenylphosphine)rhodium(I)chloride (Wilkinson's catalyst) in THF at 80 psi for several days. The lactone ring can then be converted to ether (oxepin) derivatives by procedures described in Reaction Schemes B and C. The C4 acetate group position can be modified by procedures described in Reaction Scheme G.

WO 97/16182 WO 9716182PCTIUS96/1 7482 41 REACTION SCHEME J 'OAc GAo 2> Ac I [0] 12 H 1 8 GAc 2 R 4 I (AcO) 3 B H 4" Ac bAc 94Ac

R

WO 97/16182 PCT/US96/17482 -42- The C20-29 olefin can be selectively converted to the corresponding ketone by a variety of oxidative cleavage procedures.

Ozonization (03) at reduced temperatures, preferrably at -78 0 C in dichloromethane and methanol gives the ketone in good yield.

Alternatively, the C20 ketone can be prepared by sequential reaction with osmium tetroxide or ruthenium tetroxide and sodium periodate.

The C20 ketone can be conveniently reduced to its corresponding alcohol by using tetramethylammonium triacetoxyborohydride in a solvent such as THF at room temperature.

WO 97/16182 WO 9716182PCT/US96/17482 43 REACTION SCHEME K 'OAc O' Ac GAc t 0S0 4 Me 3

NO

GAc

BH

3 .THF, Me 3

NO

OH

29 H 19 21 0 18 1722, rOH 2

PO

2

CH

3 GAc GAc WO 97/16182 PCT/US96/17482 -44- The C20-29 olefin can be converted to the hydroxymethyl derivative. One procedure involves reaction with diborane in THF followed by oxidative workup with tri-Nmethylamine-N-oxide (Me3NO).

The C20,C29 bis-alcohol can also be prepared by use of catalytic osmium tetroxide followed by oxidative workup with a reagent such as Me3NO.

WO 97/16182 WO 9716182PCT/US96,q 7482 45 REACTION SCHEME L 'GAc 'GAc 0 2

CH

3 Ac GAc R 29 a'M 2'-P020H3 GAc The C20 ketone derivative can be reacted to produce olefins with a variety of well known olefination agents. A particularly useful WO 97/16182 PCT/US96/17482 46 reagent is the Horer-Emmons-type of olefination reagent This produces a mixture of geometric isomers.

Alternatively, the C20 ketone can be reacted with nucleophiles

(R

2 9 to give C20 substituted hydroxy derivatives.

In general, Grignard reagents (R 2 9 a'MgBr) or alkyllithium reagents

(R

2 9 a'Li) are utilized in aprotic solvents such as diethyl ether or THF.

It should be understood that R 2 9a' and R29b' are used to represent a portion of the R 2 9a and R29b definitions, e.g. R 2 9a can be a substituted olefin which is depicted in the scheme as R' and R" representing alkyl substituents.

REACTION SCHEME M

OH

19 21 12 18 22 0 1 O 11 17 3b10 8' 15 OAC X b52 7 26 0 24 6 O A c OAc OAc

R

4 OR29a 12 H18 22 0 11 1 7 H H i3 H "PO2CH3 OAc 4 OAc

R

4 WO 97/16182 PCT/US96/17482 -47- REACTION SCHEME M (CONT) OAc OAc OAc 'OAc OAc The C20 hydroxy derivative (Reaction Scheme K) or the C20 hydroxymethyl derivative (Reaction Scheme L) can be converted to ether, ester, carbonate, carbamate, sulfonate and other related derivatives by procedures commonly practiced in the art and as described in Reaction Scheme G.

WO 97/16182 WO 9716182PCTIUS96/17482 48 REACTION SCHEME N OAc IR'R 2

NH

NaCNBH 3 'OAc OAc Amines at C20 can be prepared from the C20 ketone by reaction with an amnine R I R 2 NI- in a variety of solvents with a reducing agent such as sodium cyanoborohydride.

WO 97/16182PCJS6I78 PCTIUS96/17482 49 REACTION SCHEME 0

OH

12 H1 2 P 020H 3 QAc OAc WO 97/16182 WO 9716182PCT/US96 17482 50 REACTION SCHEME 0 (CONT' ,OR 29 a' OAc 2

PO

2

CH

3 'GAc OAc QAc

R

1 R 2 'OAc QAc The C20 hydroxy derivative (Reaction Scheme J) or the C20 hydroxymethyl derivative (Reaction Scheme K) can be converted to WO 97/16182 PCTIUS96/1 7482 -51 ether, ester, carbonate, carbamate, sulfonate and other related derivatives by procedures commonly practiced in the art and as described in Reaction Scheme G.

The C20 hydroxymethyl derivative may also be derivatized as a methanesulfinate ester or triflate ester by standard procedures. The methansulfinate or triflate can then be reacted with an amine NR 1

R

2

H

to give amine derivatives.

UTILITY

The present invention is related to compounds of formula I, including but not limited to those specified in the examples, which are useful in a mammalian subject for the treatment and prevention of immunemediated diseases such as the resistance by transplantation of organs or tissue such as heart, kidney, liver, medulla ossium, skin, corea, lung, pancreas, intestinum tenue, limb, muscle, nervus, duodenum, small-bowel, pancreatic-islet-cell, including xeno transplants, etc.; graft-versus-host diseases brought about by medulla ossium transplantation; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes uveitis, juvenileonset or recent-onset diabetes mellitus, posterior uveitis, allergic encephalomyelitis, glomerulonephritis, and the like; and further infectious diseases caused by pathogenic microorganisms. Further uses may include the treatment and prophylaxis of inflammatory and hyperproliferative skin diseases and cutaneous manifestations of immunologically mediated illnesses, such as psoriasis, atopical dermatitis, contact dermatitis and further eczematous dermatitises and further eczematous dermatitises, seborrhoeis dermatitis, Lichen planus, Pemphigus, bullous pemphigoid, Epidermolysis bullosa, urticaria, angioedemas, vasculitides, erythemas, cutaneous eosinophilias, Lupus erythematosus, acne and Alopecia areata; various eye diseases (autoimmune and otherwise) such as keratoconjunctivitis, vemal conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic WO 97/16182 PCT/US96/17482 -52keratitis, conical cornea, dystrophia epithelialis comeae, corneal leukoma, ocular pemphigus, Mooren's ulcer, Scleritis, Graves' opthalmopathy, Vogt-Koyanagi-Harada syndrome, sarcoidosis, etc.; pollen allergies, reversible obstructive airway disease, which includes condition such as asthma (for example, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma and dust asthma), particularly chronic or inveterate asthma (for example, late asthma and airway hyper-responsiveness), bronchitis and the like; inflammation of mucous and blood vessels such as gastric ulcers, vascular damage caused by ischemic diseases and thrombosis, ischemic bowel diseases, inflammatory bowel diseases, necrotizing enterocolitis, intestinal lesions associated with thermal bums and leukotriene B4-mediated diseases; intestinal inflammations/allergies such as Coeliac diseases, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease and ulcerative colitis; food-related allergic diseases which have symptomatic manifestation remote from the gastrointestinal tract migraine, rhinitis and eczema); renal diseases such as interstitial nephritis, Goodpasture's syndrome, hemolytic-uremic syndrome and diabetic nephropathy; nervous diseases such as multiple myositis, Guillain-Barre syndrome, Meniere's disease, polyneuritis, multiple neuritis, mononeuritis and radiculopathy; endocrine diseases such as hyperthyroidism and Basedow's disease; hematic diseases such as pure red cell aplasia, aplastic anemia, hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia and anerythroplasia; bone diseases such as osteoporosis; respiratory diseases such as sarcoidosis, fibroid lung and idiopathic interstitial pneumonia; skin disease such as dermatomyositis, leukoderma vulgaris, ichthyosis vulgaris, photoallergic sensitivity and cutaneous T cell lymphoma; circulatory diseases such as arteriosclerosis, atherosclerosis, aortitis syndrome, polyarteritis nodosa and myocardosis; collagen diseases such as scleroderma, Wegener's granuloma and Sjogren's syndrome; adiposis; eosinophilic fascitis; periodontal disease such as lesions of gingiva, periodontium, alveolar bone and substantia ossea dentis; WO 97/16182 PCT/US96/17482 -53nephrotic syndrome such as glomerulonephritis; male pattern alopecia or alopecia senilis by preventing epilation or providing hair germination and/or promoting hair generation and hair growth; muscular dystrophy; Pyoderma and Sezary's syndrome; Addison's disease; active oxygenmediated diseases, as for example organ injury such as ischemiareperfusion injury of organs (such as heart, liver, kidney and digestive tract) which occurs upon preservation, transplantation or ischemic disease (for example, thrombosis and cardiac infraction): intestinal diseases such as endotoxin-shock, pseudomembranous colitis and colitis caused by drug or radiation; renal diseases such as ischemic acute renal insufficiency and chronic renal insufficiency; pulmonary diseases such as toxinosis caused by lung-oxygen or drug (for example, paracort and bleomycins), lung cancer and pulmonary emphysema; ocular diseases such as cataracta, siderosis, retinitis, pigmentosa, senile macular degeneration, vitreal scarring and coreal alkali bum; dermatitis such as erythema multiforme, linear IgA ballous dermatitis and cement dermatitis; and others such as gingivitis, periodontitis, sepsis, pancreatitis, diseases caused by environmental pollution (for example, air pollution), aging, carcinogenis, metastasis of carcinoma and hypobaropathy; disease caused by histamine or leukotriene-C4 release; Behcet's disease such as intestinal-, vasculo- or neuro-Behcet's disease, and also Behcet's which affects the oral cavity, skin, eye, vulva, articulation, epididymis, lung, kidney and so on. Furthermore, the compounds of the invention are useful for the treatment and prevention of hepatic disease such as immunogenic diseases (for example, chronic autoimmune liver diseases such as the group consisting of autoimmune hepatitis, primary biliary cirrhosis and sclerosing cholangitis), partial liver resection, acute liver necrosis necrosis caused by toxin, viral hepatitis, shock, or anoxia), B-virus hepatitis, non-A/non-B hepatitis, cirrhosis (such as alcoholic cirrhosis) and hepatic failure such as fulminant hepatic failure, late-onset hepatic failure and "acute-onchronic" liver failure (acute liver failure on chronic liver diseases), and moreover are useful for various diseases because of their useful activity such as augmention of chemotherapeutic effect, preventing or treating WO 97/16182 PCT/US96/17482 -54activity of cytomegalovirus infection, particularly HCMV infection, and antiinflammatory activity; and The compounds of the present invention may also be used in the treatment of immunodepression or a disorder involving immunodepression, such as AIDS, cancer, senile dementia, trauma (including wound healing, surgery and shock) chronic bacterial infection, and certain central nervous system disorders.

A method of treating a condition in a mammal, the treatment of which is effected or facilitated by K 1.3 inhibition, comprising the administration, in an amount that is effective at inhibiting Ky 1.3, of a compound of Formula I. The method of treating a condition in a mammal, the treatment of which is effected or facilitated by Kv1.3 inhibition, wherein the condition is selected from the group consisting of: immunemediated diseases such as the resistance by transplantation of organs or tissue such as heart, kidney, liver, medulla ossium, skin, cornea, lung, pancreas, intestinum tenue, limb, muscle, nervus, duodenum, small-bowel, pancreatic-islet-cell, including xeno transplants, etc.; graft-versus-host diseases brought about by medulla ossium transplantation; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes uveitis, juvenileonset or recent-onset diabetes mellitus, posterior uveitis, allergic encephalomyelitis, glomerulonephritis, and the like; and further infectious diseases caused by pathogenic microorganisms. Further uses may include the treatment and prophylaxis of inflammatory and hyperproliferative skin diseases and cutaneous manifestations of immunologically mediated illnesses, such as psoriasis, atopical dermatitis, contact dermatitis and further eczematous dermatitises and further eczematous dermatitises, seborrhoeis dermatitis, Lichen planus, Pemphigus, bullous pemphigoid, Epidermolysis bullosa, urticaria, angioedemas, vasculitides, erythemas, cutaneous eosinophilias, Lupus erythematosus, acne and Alopecia areata; various eye diseases (autoimmune and otherwise) such as keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic WO 97/16182 PCT/US96/17482 keratitis, conical cornea, dystrophia epithelialis comeae, corneal leukoma, ocular pemphigus, Mooren's ulcer, Scleritis, Graves' opthalmopathy, Vogt-Koyanagi-Harada syndrome, sarcoidosis, etc.; pollen allergies, reversible obstructive airway disease, which includes condition such as asthma (for example, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma and dust asthma), particularly chronic or inveterate asthma (for example, late asthma and airway hyper-responsiveness), bronchitis and the like; inflammation of mucous and blood vessels such as gastric ulcers, vascular damage caused by ischemic diseases and thrombosis, ischemic bowel diseases, inflammatory bowel diseases, necrotizing enterocolitis, intestinal lesions associated with thermal bums and leukotriene B4-mediated diseases; intestinal inflammations/allergies such as Coeliac diseases, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease and ulcerative colitis; food-related allergic diseases which have symptomatic manifestation remote from the gastrointestinal tract migraine, rhinitis and eczema); renal diseases such as interstitial nephritis, Goodpasture's syndrome, hemolytic-uremic syndrome and diabetic nephropathy; nervous diseases such as multiple myositis, Guillain-Barre syndrome, Meniere's disease, polyneuritis, multiple neuritis, mononeuritis and radiculopathy; endocrine diseases such as hyperthyroidism and Basedow's disease; hematic diseases such as pure red cell aplasia, aplastic anemia, hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia and anerythroplasia; bone diseases such as osteoporosis; respiratory diseases such as sarcoidosis, fibroid lung and idiopathic interstitial pneumonia; skin disease such as dermatomyositis, leukoderma vulgaris, ichthyosis vulgaris, photoallergic sensitivity and cutaneous T cell lymphoma; circulatory diseases such as arteriosclerosis, atherosclerosis, aortitis syndrome, polyarteritis nodosa and myocardosis; collagen diseases such as scleroderma, Wegener's granuloma and Sjogren's syndrome; adiposis; eosinophilic fascitis; periodontal disease such as lesions of gingiva, periodontium, alveolar bone and substantia ossea dentis; WO 97/16182 PCT/US96/17482 -56nephrotic syndrome such as glomerulonephritis; male pattern aleopreia or alopecia senilis by preventing epilation or providing hair germination and/or promoting hair generation and hair growth; muscular dystrophy; Pyoderma and Sezary's syndrome; Addison's disease; active oxygenmediated diseases, as for example organ injury such as ischemiareperfusion injury of organs (such as heart, liver, kidney and digestive tract) which occurs upon preservation, transplantation or ischemic disease (for example, thrombosis and cardiac infraction): intestinal diseases such as endotoxin-shock, pseudomembranous colitis and colitis caused by drug or radiation; renal diseases such as ischemic acute renal insufficiency and chronic renal insufficiency; pulmonary diseases such as toxinosis caused by lung-oxygen or drug (for example, paracort and bleomycins), lung cancer and pulmonary emphysema; ocular diseases such as cataracta, siderosis, retinitis, pigmentosa, senile macular degeneration, vitreal scarring and coreal alkali bum; dermatitis such as erythema multiforme, linear IgA ballous dermatitis and cement dermatitis; and others such as gingivitis, periodontitis, sepsis, pancreatitis, diseases caused by environmental pollution (for example, air pollution), aging, carcinogenis, metastasis of carcinoma and hypobaropathy; disease caused by histamine or leukotriene-C4 release; Behcet's disease such as intestinal-, vasculo- or neuro-Behcet's disease, and also Behcet's which affects the oral cavity, skin, eye, vulva, articulation, epididymis, lung, kidney and so on. Furthermore, the compounds of the invention are useful for the treatment and prevention of hepatic disease such as immunogenic diseases (for example, chronic autoimmune liver diseases such as the group consisting of autoimmune hepatitis, primary biliary cirrhosis and sclerosing cholangitis), partial liver resection, acute liver necrosis necrosis caused by toxin, viral hepatitis, shock, or anoxia), B-virus hepatitis, non-A/non-B hepatitis, cirrhosis (such as alcoholic cirrhosis) and hepatic failure such as fulminant hepatic failure, late-onset hepatic failure and "acute-onchronic" liver failure (acute liver failure on chronic liver diseases), and moreover are useful for various diseases because of their useful activity such as augmention of chemotherapeutic effect, preventing or treating WO 97/16182 PCT/US96/17482 -57activity of cytomegalovirus infection, particularly HCMV infection, and antiinflammatory activity; and immunodepression or a disorder involving immunodepression, such as AIDS, cancer, senile dementia, trauma (including wound healing, surgery and shock), chronic bacterial infection, and certain central nervous system disorders.

An embodiment of the invention is a method for the treatment of autoimmune diseases. Another embodiment of the invention is a method for the prevention of rejection of foreign organ transplants comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula I.

One end result of an autoimmune or a rejection process is tissue destruction caused by inflammatory cells and the mediators they release. Anti-inflammatory agents such as NSAID's and corticosteroids act principally by blocking the effect or secretion of these mediators, but do nothing to modify the immunologic basis of the disease. On the other hand, cytotoxic agents, such as cyclophosphamide, act in such a nonspecific fashion that both the normal and autoimmune responses are shut off. Indeed, patients treated with such nonspecific immunosuppressive agents are as likely to succumb from infection as they are from their autoimmune disease.

Cyclosporin A, which was approved by the US FDA in 1983, is currently the leading drug used to prevent rejection of transplanted organs. The drug acts by inhibiting the body's immune system from mobilizing its vast arsenal of natural protecting agents to reject the transplant's foreign protein. Though cyclosporin A is effective in fighting transplant rejection, it is nephrotoxic and is known to cause several undesirable side effects including kidney failure, abnormal liver function and gastrointestinal discomfort.

Newer, safer drugs exhibiting fewer side effects are constantly being searched for in the field. The present invention provides for immunosuppressant agents which are inhibitors of a voltage dependent potassium channel, Kv1.3, that is found on human T-lymphocytes.

WO 97/16182 PCT/US96/17482 58 Potassium channels modulate a number of cellular events such as muscle contraction, neuro-endocrine secretion, frequency and duration of action potentials, electrolyte homeostasis, and resting membrane potential. These channels comprise a family of proteins that have been classified according to their biophysical and pharmacological characteristics. Inhibition of K+ channels, in their role as modulators of the plasma membrane potential in human T -lymphocytes, has been postulated to play a role in eliciting immunosuppressive responses. In regulating membrane potential, K+ channels play a role in the regulation of intracellular Ca++ homeostasis, which has been found to be important in T-cell activation. The biochemical characterization of K+ channels is underdeveloped, due to the paucity of selective high affinity probes.

Functional voltage-gated K+ channels can exist as multimeric structures formed by the association of either identical or dissimilar subunits. This phenomena is thought to account for the wide diversity of K+ channels. However, subunit compositions of native K+ channels and the physiologic role that particular channels play are, in most cases, still unclear.

The Kv 1.3 channel is a voltage-gated potassium channel that is found in neurons, blood cells, osteoclasts and T-lymphocytes.

The Chandy and Cahalan laboratories proposed a hypothesis that blocking the Kv1.3 channel would elicit an immunosuppressant response. (Chandy et al., J. Exp. Med. 160, 369, 1984; Decoursey et al., Nature, 307, 465, 1984). However, the K+ channel blockers employed in their studies were non-selective. Until research with the peptide margatoxin, a peptide found in scorpion venom, no specific inhibitor of the Kv 1.3 channel existed to test this hypothesis. Although a laboratory (Price et al., Proc. Natl. Acad. Sci. USA, 86, 10171, 1989) showed that charybdotoxin would block Kvl.3 in human T cells, charybdotoxin was subsequently shown to inhibit four different K+ channels (Kv1.3 and three distinct small conductance Ca++ activated K+ channels) in human T-lymphocytes, limiting the use of this toxin as a probe for the physiological role of Ky 1.3 (Leonard et al., Proc. Natl. Acad. Sci. USA, WO 97/16182 PCT/US96/17482 -59- 89, 10094, 1992). Margatoxin, on the other hand, blocks only Kvl.3 in T-cells, and has immunosuppressant activity in both in vitro and in vivo models. (Lin et al., J. Exp. Med, 177, 637, 1993). Since the compounds of the embodiments of this invention produce blockade of Ky 1.3, they will also inhibit T-cell activation.

Also within the scope of this invention is a method of treating a condition in a mammal, the treatment of which is effected or facilitated by K 1.3 inhibition, comprising the administration of a pharmaceutical composition comprising a suitable pharmaceutical carrier and a compound of Formula in an amount that is effective at inhibiting Kv1.3.

Also within the scope of this invention is a combination therapy comprising a compound of formula I and one or more immunosuppressant agents. These immunosuppressant agents within the scope of this invention include, but are not limited to, IMUREK® azathioprine sodium, brequinar sodium, SPANIDIN® gusperimus trihydrochloride (also known as deoxyspergualin), mizoribine (also known as bredinin), CELLCEPT® mycophenolate mofetil, NEORAL® Cyclosporin A (also marketed as different formulation of Cyclosporin

A

under the trademark SANDIMMUNE®), PROGRAF® tacrolimus (also known as FK-506) and RAPIMMUNE® sirolimus (also known as rapamycin), leflunomide (also known as HWA-486), glucocortcoids, such as prednisolone and its derivatives, antibody therapies such as orthoclone (OKT3) and Zenapax and antithymyocyte globulins, such as thymoglobulins.

Using the methodologies described below, representative compounds of the invention were evaluated and found to exhibit values of at least <10 gM in any of the assays thereby demonstrating and confirming the utility of the compounds of the invention as Kv1.3 inhibitors and immunosuppressants.

T CELL IL-2 ASSAY Peripheral blood mononuclear (MNC) cells from healthy donors were separated by density centrifugation with ficoll-hypaque WO 97/16182 PCT/US96/17482 (LSM, Organon Teknika, Durham, NC), followed by rosetted with neuraminidase treated sheep red blood cells (SRBC). After another centrifugation with leucocyte separation medium (LSM), the SRBC of the rosetted T cells were then lysed with ammonium chloride lysing buffer (GIBCO, Grand Island, NY). Such purified T cells were resuspended at 3 X 106/ ml in RPMI 1640 culture medium (GIBCO) supplemented with 10% fetal calf serum (Sigma, St. Louis, MO), 100 mM glutamine, 1 mM sodium pyruvate, 0.1 mM non-essential amino acids, and I penn-strep (GIBCO). The cell suspension was immediately distributed into 96 well round-bottom microculture plates (Costar) at 200 kl/well. The various dilutions of test compound were then added in triplicate wells at 25 pl/well, incubated for 30 min at 37°C. lonomycin (125 ng/ml), and PMA (1 or 5 ng/ml), were added to the appropriate wells. The culture plates were then incubated at 37 0 C in a humidified atmosphere of 5% C02 95% air for 18-24 hours. The supematants were removed, and assayed for IL-2 with an IL-2 capture ELISA, using monoclonal anti-IL-2, and biotinylated goat anti-IL-2 antibodies (unconjugated antibodies purchased from R&D System, Minneapolis, MN). The ELISA was developed with streptavidin conjugated peroxidase (Zymed, San Francisco, CA) and substrate for peroxidase (Sigma). Mean OD and units of IL-2 of the replicate wells were calculated from standard curve, created with recombinant IL-2 (Collaborative Biomedical Products, Bedford, MA) and the results were expressed as concentration of compound required to inhibit IL-2 production of T cells by T CELL PROLIFERATION

ASSAY

Peripheral blood mononuclear cells (MNC) from healthy donors were separated by density centrifugation with ficoll-hypaque (LSM, Organon Teknika, Durham, NC). After washing the MNC with complete media (RPMI 1640 medium with 5% fetal calf serum, 100 mM glutamine, 1 mM sodium pyruvate, 0.1 mM non-essential amino acid, and 1% penn-strep, obtained from GIBCO, Grand Island, NY), they were then irradiated at 7500 RADS, and resuspended at 4-4.5 x WO 97/16182 PCT/US96/17482 -61 106cells/ml in complete media. Another aliquot of MNC were rosetted with neuraminidase treated SRBC. After another centrifugation with LSM, the sheep red blood cells (SRBC) of these rosetted T cells were then lysed with ammonium chloride lysing buffer (GIBCO, Grand Island, NY). After washing 2X with complete media, these purified T cells were also resuspended at 2-2.5 x 106cells/ml in complete media.

The various dilutions of the compound were added in triplicates at ul/well of a 96 well flat-bottom microculture plate (Costar, Cambridge, MA). T cell suspension was then immediately distributed into the wells at 100 ul/well. After incubating the cells with compound for 30 min. at 37 0 C in a humidified atmosphere of 5% C02 95% air, 20 gl/well of anti-CD3 (Ortho Diagnostic, NJ) at final conc. of 0.3 ng/ml was added, followed by 50 gtl of the irradiated MNC. The culture plates were then incubated at 37C in a humidified atmosphere of 5% C02 95% air for 72 hours. The proliferation of T lymphocytes was assessed by measurement of tritiated thymidine incorporation. During the last 18- 24 hrs. of culturing, the cells were pulse-labeled with 2 pCi/well of tritiated thymidine (NEN, Cambridge, MA). The cultures were harvested on glass fiber filters using a multiple sample harvester (MACH-II, Wallac,Gaithersburg, MD). Radioactivity of filter discs corresponding to individual wells was measured by standard liquid scintillation counting methods (Betaplate Scint Counter, Wallac). Mean counts per minute of replicate wells were calculated and the results were expressed as concentration of compound required to inhibit tritiated thymidine uptake of T cells by KV1.3-RUBIDIUM EFFLUX ASSAY CHO cells transfected with Ky 1.3 channels at site densities of approximately 40,000 sites/cell are plated into 96 well culture plates and maintained in Iscove's Modified Dulbecco's Medium (IMDM, with L-Glutamine and HEPES, JRH Biosciences). Cells are incubated overnight with 86Rb+ (3 tCi/ml, Dupont-NEN) in the glutamine supplemented IMDM. After aspiration of the media, 100 [tl of Low K Buffer (in mM, 6.5 KC1, 125 NaCI, 1 CaCl2, 2 MgCl2, 10 HEPES, pH WO 97/16182 PCT/US96/17482 -62adjusted to 7.2 with NaOH) is added to each well followed by 100 gl test samples in Low K Buffer also containing 0.2% BSA and 2 mM ouabain.

Samples are tested at either 1 gg/ml for routine screening or at a variety of concentrations encompassing at least 1/10 to 10 times the putative

IC

5 0 of test compound to determine potency. After a fixed preincubation time, which is usually 10 min, the samples are aspirated.

The Kv1.3 channels are opened by depolarization of the cells with High K Buffer (final concentrations, in mM, 63.25 KC1, 68.25 NaCl, 1 CaCl2, 2 MgCl2, 10 HEPES, pH adjusted to 7.2 with NaOH) also containing test compounds. To measure 8 6 Rb+ efflux through the channels, aliquots of 100 gl are taken from each well after a given time and added to plates containing 100 gl MicroScint-40 (Packard) for counting by liquid scintillation techniques. MicroScint-40 (100 gl) is then added to each well of the cell plate to determine the remaining 86 Rb+ activity. The efflux counts are normalized for the total amount of 8 6 Rb+ that was in the cells by adding the efflux counts to the cell plate counts. Activity is determined by inhibition of the efflux window that is established using a saturating concentration of margatoxin (MgTX), a 39 amino acid peptide that is a potent blocker of K 1.3 channels (IC 5 0 100 pM).

DOSAGE FORMS As an immunosuppressive, these compounds are useful in the treatment of autoimmune diseases, the prevention of rejection of foreign organ transplants and/or related afflictions, diseases and illnesses.

The compounds of this invention can be administered for the treatment of autoimmune diseases, the prevention of rejection of foreign organ transplants and/or related afflictions, diseases and illnesses according to the invention by any means that effects contact of the active ingredient compound with the site of action in the body of a warmblooded animal. For example, administration, can be oral, topical, including transdermal, ocular, buccal, intranasal, inhalation, intravaginal, rectal, intracisteral and parenteral. The term WO 97/16182 PCT/US96/17482 -63- "parenteral" as used herein refers to modes of administration which include subcutaneous, intravenous, intramuscular, intraarticular injection or infusion, intrastemrnal and intraperitoneal.

The compounds can be administered by any conventional means available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in a combination of therapeutic agents.

They can be administered alone, but are generally administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.

For the purpose of this disclosure, a warm-blooded animal is a member of the animal kingdom possessed of a homeostatic mechanism and includes mammals and birds.

The dosage administered will be dependent on the age, health and weight of the recipient, the extent of disease, kind of concurrent treatment, if any, frequency of treatment and the nature of the effect desired. Usually, a daily dosage of active ingredient compound will be from about 1-500 milligrams per day. Ordinarily, from 10 to 100 milligrams per day in one or more applications is effective to obtain desired results. These dosages are the effective amounts for the treatment of autoimmune diseases, the prevention of rejection of foreign organ transplants and/or related afflictions, diseases and illnesses.

The active ingredient can be administered orally in solid dosage forms, such as capsules, tablets, troches, dragees, granules and powders, or in liquid dosage forms, such as elixirs, syrups, emulsions, dispersions, and suspensions. The active ingredient can also be administered parenterally, in sterile liquid dosage forms, such as dispersions, suspensions or solutions. Other dosages forms that can also be used to administer the active ingredient as an ointment, cream, drops, transdermal patch or powder for topical administration, as an ophthalmic solution or suspension formation, eye drops, for ocular administration, as an aerosol spray or powder composition for inhalation or intranasal administration, or as a cream, ointment, spray or suppository for rectal or vaginal administration.

WO 97/16182 PCT/US96/17482 -64- Gelatin capsules contain the active ingredient and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.

Liquid dosage forms for oral administration can contain coloring and flavoring to increase patient acceptance.

In general, water, a suitable oil, saline, aqueous dextrose (glucose), and related sugar solutions and glycols such as propylene glycol or polyethylene gycols are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain a water soluble salt of the active ingredient, suitable stabilizing agents, and if necessary, buffer substances. Antioxidizing agents such as sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or combined, are suitable stabilizing agents. Also used are citric acid and its salts and sodium EDTA. In addition, parenteral solutions can contain preservatives, such as benzalkonium chloride, methyl- or propylparaben, and chlorobutanol.

Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, A. Osol, a standard reference text in this field.

For administration by inhalation, the compounds of the present invention may be conveniently delivered in the form of an aerosol spray presentation from pressurized packs or nebulisers. The compounds may also be delivered as powders which may be formulated and the powder composition may be inhaled with the aid of an insufflation powder inhaler device. The preferred delivery system for inhalation is a metered dose inhalation (MDI) aerosol, which may be formulated as a suspension or solution of a compound of Formula I in suitable propellants, such as fluorocarbons or hydrocarbons.

'I

WO 97/16182 PCT/US96/17482 For ocular administration, an ophthalmic preparation may be formulated with an appropriate weight percent solution or suspension of the compounds of Formula I in an appropriate ophthalmic vehicle, such that the compound is maintained in contact with the ocular surface for a sufficient time period to allow the compound to penetrate the corneal and internal regions of the eye.

Useful pharmaceutical dosage-forms for administration of the compounds of this invention can be illustrated as follows:

CAPSULES

A large number of unit capsules are prepared by filling standard two-piece hard gelatin capsules each with 100 milligrams of powdered active ingredient, 150 milligrams of lactose, 50 milligrams of cellulose, and 6 milligrams magnesium stearate.

SOFT GELATIN CAPSULES A mixture of active ingredient in a digestible oil such as soybean oil, cottonseed oil or olive oil is prepared and injected by means of a positive displacement pump into gelatin to form soft gelatin capsules containing 100 milligrams of the active ingredient. The capsules are washed and dried.

TABLETS

A large number of tablets are prepared by conventional procedures so that the dosage unit is 100 milligrams of active ingredient, 0.2 milligrams of colloidal silicon dioxide, 5 milligrams of magnesium stearate, 275 milligrams of microcrystalline cellulose, 11 milligrams of starch and 98.8 milligrams of lactose. Appropriate coatings may be applied to increase palatability or delay absorption.

WO 97/16182 PCT/US96/17482 -66-

INJECTABLE

A parenteral composition suitable for administration by injection is prepared by stirring 1.5% by weight of active ingredient in 10% by volume propylene glycol. The solution is made to volume with water for injection and sterilized.

SUSPENSION

An aqueous suspension is prepared for oral administration so that each 5 milliliters contain 100 milligrams of finely divided active ingredient, 100 milligrams of sodium carboxymethyl cellulose, milligrams of sodium benzoate, 1.0 grams of sorbitol solution, U.S.P., and 0.025 milliliters of vanillin.

The same dosage forms can generally be used when the compounds of this invention are administered stepwise or in conjunction with another therapeutic agent. When drugs are administered in physical combination, the dosage form and administration route should be selected depending on the compatibility of the combined drugs. Thus the term coadministration is understood to include the administration of the two agents concomitantly or sequentially, or alternatively as a fixed dose combination of the two active components.

The following examples illustrate the preparation of the compounds of Formula I and as such are not to be considered as limiting the invention set forth in the claims appended hereto.

WO 97/16182 PCT/US96/17482 -67- EXAMPLE I A Method Of Extracting The Compounds Of Formula 1(a) and 1(b) From Spachea correa H H H H C H bond and R is OAc OH COOCH3 O CH" CH3 OAc Formula 1(b) b is a double 0 OAc bond and R is OAc OAc CH OAc O Ac

OR

One gram of an ethanol extract of the roots of Spachea correa was partitioned between 100 ml of hexane (twice) and 100 ml of 90% aqueous methanol. After separation of the phases, the defatted methanol was concentrated down under vacuum to give an aqueous suspension. This was diluted out to 100 ml with water and extracted, with 100 ml of methylene chloride.

The bioactive methylene chloride extract was dried down to give 12 mg of residue. This was first fractionated by preparative thin layer chromatography (TLC) on a 20 cm by 20 cm E. Merck silica gel 60F254 plate of 1mm thickness using methylene chloride-ethyl acetate 1:1 as solvent, then by high performance liquid chromatography (HPLC) using a Zorbax RxC8 4.6 mm x 25 cm column, operated at 0 C and eluted with a 50 minute gradient of acetonitrile:water (1:1, v/v) to 100% acetonitrile, delivered at 1 ml/min, to afford 4 mg of compound 1(a) and 1 mg of 1(b).

Homogeneity of the preparations was ascertained in several TLC systems, such as E. Merck silica gel 60F254, methylene chlorideethyl acetate 1:1, Rf 1(a) 0.4, Rf 1(b) 0.3; Whatman KCl8, methanolwater 9:1, Rf 1(a) 0.65, Rf 1(b) 0.75 and by HPLC using a Zorbax WO 97/16182 PCT/US96/17482 -68 RxC8 column, acetonitrile-water 3:2, k' l(a) 4.15, k' 1(b) 3.30; and by

NMR.

Mass spectra were recorded on JEOL SX-102A (electron impact, EI,903V) and JEOL HX110 (Fast Atom Bombardment,

FAB)

mass spectrometers. Exact mass measurements were performed at high resolution (HR-EI) using perfluorokerosene (PFK) as the internal standard. Trimethylsilyl derivatives were prepared with a 1:1 mixture of BSTFA-pyridine at room temperature The FAB spectrum was run in a matrix of dithiothreitol (20/80).

The compound of Formula 1(a) runs underivatized by EI.

The molecular ion is observed a m/z 788 and three successive loses of acetic acid are observed. The base peak is observed a m/z 334. The compound does not silylate. Scanning HR-EI indicated a molecular formula of C40H52016. A table of the critical HR-EI data is given below.

Observed m/z Formula Assignment 788.3220 C40H52016

M+

728.3040 C38H48014 M-acetic acid 668.2834 C36H44012 M-2 x acetic acid 334.1417 C18H2206 base peak 13 C NMR spectra were recorded for the compound of Formula 1(a) in CD2C12 at 100 MHz on a Varian Unity 400 NMR spectrometer at 20 0 C. Chemical shifts are given in ppm relative to tetramethylsilane (TMS) at zero ppm using the solvent peak at 53.8 ppm as internal standard. The following data were observed: 15.0, 15..2, 16.8, 17.1, 20.7*, 20.9, 21.1, 21.6, 21.8, 22.2, 35.6, 40.8*, 42.1, 43.6, 45.1, 47.5, 49.3*, 53.5, 59.1, 62.6, 63.5, 66.1, 66.7*, 68.4*, 69.9, 73.9, 75.0, 75.6, 77.1*, 119.4, 123.7, 138.9, 143.0, 167.7, 169.2, 169.3*, 170.25, 170.31, 170.8, 171.3 ppm (where the signifies the observation as broad resonances). The carbon count of 40 is in agreement with the molecular formula C40H520 16 derived by scanning HR EI-MS.

WO 97/16182 PCT/US96/17482 -69- The 1H NMR spectra of compound of Formula(a) is provided as Figure 1. The spectra was recorded at 400 MHz in CD2C12 on a Varian Unity 400 NMR spectrometer at 25 0 C. Chemical shifts are in ppm relative to TMS at zero ppm using the solvent peak at 85.32 as the internal standard.

The mass spectra of the compound of Formula 1(b) was obtained as above. The following results were obtained.

Observed m/z Formula Assignment 786.3075 C40H50016

M+

726.2886 C38H46014 M-acetic acid 666.2651 C36H42012 M-2 x acetic acid 606.2451 C34H38010 M-3 x acetic acid 489.2099 C26H3309 base peak 471.1992 C26H3108 13 C NMR spectra were recorded for the compound of Formula l(b) using the procedure described above. The following results were observed: 14.8, 14.9, 17.3, 20.8, 20.9, 21.3, 21.7, 21.8, 21.9, 27.1, 35.1, 40.6, 42.3, 45.4, 48.1, 50.4, 53.5, 54.1, 57.8, 63.7, 66.2, 67.8, 68.6, 71.4, 73.3, 73.8, 74.4, 119.5, 121.1, 124.3, 137.1, 138.9, 143.3, 167.6, 168.6, 169.3, 169.5, 169.9, 171.0, 171.7 ppm.

The carbon count of 40 is in agreement with the molecular formula C40H50016 derived by scanning HR El-MS.

WO 97/16182 PCT/US96/17482 EXAMPLE 2 A Method Of Extracting The Compounds Of Formula 1(c) And 1(d) From Spachea Correa H H H,,,1H ,H b CH 0 Formula 1(c) b is a single H bond and R is OH OH COOCH3 0I CH CHo OAc Formula 1(d) b is a double O O\ c bond and R is OH °Ac CHq OAc

OR

Analogs of the compounds of Formula 1(a) and 1(b) could be detected in the crude extract and fractions thereof when the process of Example 1 was carried out on a larger scale. Thus, 50 g of ethanol extract were partitioned as described in Example 1 using 900 ml of each solvent at each step.

Partial purification of the methylene chloride extract was achieved by column chromatography on E. Merck silica gel 60 (120 ml), eluting with a step gradient of ethyl acetate in methylene chloride.

The step gradient was designed so that the column was washed first with 100% methylene chloride and then with methylene chloride- ethyl acetate mixtures of 9:1, 8:2, 3:2, 2:1, 1:1, 1:2, 2:8 and 1:9. Ultimately the column was washed with 100% ethyl acetate. Fractions eluted with methylene chloride-ethyl acetate 3:2 were enriched in compound of Formula 1(a) and These were resolved by HPLC using a Zorbax RxC8 9 mm x 25 cm column, maintained at 50°C and eluted at 4 ml/min with acetonitrile-water 1:1 v/v. Three identical runs finally afforded 100 mg and 20 mg respectively of l(a) and 1(b) after crystallization from methanol. Later-eluting fractions from the silica gel column above were found to contain at least two related compounds based on UV spectra and color reactions on TLC plates. Material from the WO 97/16182 PCT/US96/17482 -71 methylene chloride-ethyl actate 1:1 and 1:2 washings were combined and evaporated down. Separation was achieved on the same HPLC column as above, eluting with a 50 minute gradient of 30% to acetonitrile in water. Two identical runs gave 6 mg of purified compound Fractions containing the compound of Formula 1(d) were again processed by HPLC (same column) using acetonitrile-water 3:7 delivered isocratically, to yield 2 mg of purified Formula 1(d).

The mass spectra of these compounds were recorded on a Finnigan TSQ700 mass spectrometer (electrospray ionization, ESI).

The samples were analyzed by LC/MS using a 2.1x150mm C8 column at 0.2ml/min. with a mobile phase of 45% acetonitrile/0.01M aqueous ammonium acetate at 50°C. Component 1(d) had a retention time of 10.5 min. and a molecular weight of 744 which is observed a m/z: 745 762 (M+NH3), 786 (M H MeCN). Component 1(c) has a retention time of 11.8 and a molecular weight of 746 which is observed at m/z: 747 764 (M+NH3) and 788 (M H MeCN).

The 1 3 C NMR spectra obtained for the compound of Formula 1(c) using the conditions previously described is as follows: 15.1 16.9, 19.8, 20.8, 20.91, 20.94, 21.9, 22.3, 35.6, 40.6, 42.2, 43.9, 45.0, 47.7, 50.8, 53.5, 55.6, 61.8, 63.5, 66.0, 67.6 69.8, 70.0, 73.9, 75.0, 75.6, 119.3, 123.7, 139.0, 144.4, 167.8, 169.2, 169.5, 170.1, 170.4, 171.4 ppm.

The carbon count of 38 is in agreement with the molecular formula C38H50016 derived by scanning HR EI-MS.

EXAMPLE 3 Separation By HPLC Compounds of this invention were characterized by the following behavior during HPLC separation on a Zorbax RxC8 4.6 mm x 25 cm column, maintained at 50 0 C and eluted at 1 ml/min with acetonitrile-water 3:2 v/v): Compound k' 4.15; k'=3.30; k'=2.30; k'=2.10.

WO 97/16182 PCT/US96/17482 -72- Analyses using this HPLC system can be used to quantify the compounds in the crude extract or other mixtures, by comparing the absorbance of HPLC peaks at a wavelength of 220 nm with that produced by injections of known (weighed) amounts of pure standards.

EXAMPLE 4 Additional Purification Procedure A simplified purification process allows for rapid fractionation of even larger amounts of crude extract and the preparation of gram amounts of the compounds of Formula 1(a) and 1(b).

The ethanol extract is first dissolved at 20 grams per 150 ml in methanol. This solution is diluted with 150 ml of water and then extracted three times with methylene chloride using 150 ml of methylene chloride each time. The pooled methylene chloride extracts are evaporated down and fractionation proceeds by repeated column chromatography on silica gel. One employs methylene chloridemethanol 97:3 in a first step; the mixed compounds of Formula 1 and 1 thus obtained are resolved by chromatographing on fresh silica gel eluted with methylene chloride-ethyl acetate 3:1. Volume of elution for the compound of Formula 1 ranges from about 2 to about 3.5 column volumes of solvent; that for the compound of Formula 1(b) is about 3 to about 4.5 column volumes. Finally, advantage is taken of the low solubility of these compounds, and, after total resolution by chromatography, the compounds of Formula 1 and 1 can be precipitated and or crystallized from concentrated methanol solutions.

WO 97/16182 PCT/US96/17482 -73- EXAMPLE 4,6,7,15,16-Pentakis(acetyloxy)-21,22-epoxy-1 8-hydroxy-22methoxycarbonyl[6a,7a, 15p, 16p,20a,21 P,22p]D:A-Friedo-A-homo- 27.30-dinor-24-oxaoleana-1 -en-3-one H OH C02 H3 0 OAc OAc 0 OAc AcO A c A solution of 50 mg (63.7 gmole) of 4,6,7,15,16- Pentakis(acetyloxy)-21,22-epoxy-18-hydroxy-22-methoxycarbonyl- [6a,7a, 15p, 160,21 P,22p]D:A-Friedo-A-homo-27,30-dinor-24oxaoleana-1,20(29)-dien-3-one in 50 ml of dry THF was degassed under reduced pressure and saturated with nitrogen, the procedure being repeated several times. 25 mg of Wilkinson's catalyst (PPh3)3RhCI were added and the solution was degassed and saturated with hydrogen in the previously described manner. The reaction vessel was then pressurized with H2 to 15 psi (latm) and shaken for 65h at 25 0 C. After that time the solvent was removed under reduced pressure. The residue was dissolved in a small amount of ethyl acetate/hexanes (ca. 1 mL) and filtered through 30g of silica gel eluting with 500 ml of ethyl acetate hexanes The first fractions, containing the Wilkinsoncatalyst (approx. 50 mL) were discarded. The fractions containing the product were combined. After removing the solvent under reduced pressure the crude product was dried in vacuo and purified by HPLC to afford 20.8 mg of the title compound as a white solid. 1H NMR (CDC13) 8 1.5 3H, J 8.6 Hz, C29-H), 2.4 1H, C20-H); Mass Spectrum (APCI): m/e 808.

WO 97/16182 PCT/US96/17482 -74- EXAMPLE 6 4,6,7,15,16-Pentakis(acetyloxy)-21,22-epoxy-1 8-hydroxy-22-methoxycarbonyl[6a,7a, 153,16p,20a,21 P,22p]D:A-Friedo-A-homo-27,30dinor-24-oxaoleana-1 -ene H 0OH

CO

2

CH

3 OAc S OAc OAc AcO/ A c A solution of 213 mg (0.27 mmole) of 4,6,7,15,16pentakis(acetyloxy)-21,22-epoxy-I 8 -hydroxy-22-methoxycarbonyl- [6a,7a,153,16p,20a,21 ,22p]D:A-Friedo-A-homo-27,30-dinor-24oxaoleana- I-en-3-one in 2 mL of dichloromethane was cooled to 0°C.

Then 0.68 mL of a 1.02M solution of lithium tri-tertbutoxyaluminum hydride was added and the solution was stirred at 0°C for 18h. The reaction was quenched with 10 mL of 2.5M aqueous H2S04 and then was diluted with 50 mL of dichloromethane. After the layers were separated, the organic phase was dried over Na2SO4 and concentrated.

The residue was dissolved in a mixture of 5 mL of dichloromethane and 1 mL of triethylsilane. Then 0.8 mL of boron trifluoride-etherate was added and the solution was stirred at room temperature. After 2 h, the reaction was quenched by addition of saturated aqueous NaHCO3 solution and dichloromethane. The layers were separated and the organic layer was washed with and brine, saturated aqueous NaHCO3, dried over MgSO4, and concentrated. The residue was purified by silica gel chromatography using S (hexane:tbutylmethylether:acetonitrile 8:4:1) to afford 60.5 mg of the title compound as a white solid; IH NMR (CDC13) 5 2.41 1H, WO 97/16182 PCT/US96/1 7482 75 3.38 IlH, C21-H), 3.69 (dd, 2H, AB, J 12 Hz, C24-H), 5.5 (in, I H, ClI-H), 5.57 (in, I1H, C2-H); Mass Spectrum (APCI): m/e 794 (M+N1-4).

EXAMPLE 7 4,6,7,15,1 6-Pentaki s(acety lox y) -20(29),2 1,22 -diep oxy I 8-hydroxy-22methoxycarbonyl[6ac,7u, 15Pf, I 6f,21 f,22f]D:A-Friedo-A-homo-27,30.

dinor-24-oxaoleana- I -en-3-one 100 cc~ A solution of 50 mg of 4 6 7 ,l5,l6-pentakis(acetyloxy)- 21,22-epoxy- I 8-hydroxy-22-methoxycarbonyl[6a,7x, 1 Sf3, 1 6f,2 1 f3,- 223] A-Friedo-A-homo-27 3 O-dinor-24-oxaoleana- 1 ,20(29)-dien-3 one (0.05 mmole) and 150 mng of m-chloro perbenzoic acid (80 remainder water; 0.7 mmole) in 5 ml of CHC13 was stored at 0 0 C for 21 days. The solution was diluted with 50 ml of dichioromethane, 20 ml of a saturated aqueous solution of NaHCO3 was added and the layers were separated. The organic layer was consecutively washed with 20 mL of 1IM hydrochloric acid and saturated aqueous sodium chloride then was dried over MgSO4 and concentrated. The residue was purified by silica gel chromatography with 1:1 ethyl acetate-hexane to afford 41.8 mg of the title compound as a white solid IH NMR (CDCl3) 6 2.92, 3.21 (AB, 2H, J 4.0 Hz, C29-H), 3.30 IJH, C2 6.09 I H, J= 11.9 Hz, C2-H), 6.33 (dd, I1H, J I 11.9 Hz, J 2 8.9Hz, ClI-H); 13

C

NMR (CDCl3) 5 123.5 142.3 Mass Spectrum (APCI): W/e 822 (M+NH4).

WO 97/16192 WO 9716182PCT/US96/17482 76 EXAMPLE 8 4,6,7,15,1 6-Pentakis(acetyloxy)-2 1,22-epoxy- I 8-hydroxy-22methoxycarbonyl [6a.,7x, 15Sf31 6f,21 1,22f]D:A-Friedo-A-homo-27,30dinor-24-oxaoleana- 1 -en-3 O~c AcO

P~

A solution of 13.7 mg of 4 ,6,7,15,16-Pentakis(acetyloxy)- 21,22-epoxy- I 8 -hydroxy-22-methoxycarbonyl[6ux,7Q, 1 5P, 1603,2 1 P3, 22f]D:A-Friedo-A-homo-27 ,30-dinor-24-oxaoleana- 1 ,20(29)-dien-3one in 12 ml of 1, CH2Cl2/CH3OH) was cooled to -78'C and 03 was bubbled into the solution until it contained a blue color. The solution was then purged with nitrogen for 3 minutes and 3 drops of Me2S was added. The solution was allowed to warm to 25'C for 14 hours.

Volatiles were removed by vacuum and the residue was purified by chromatography on silica gel using 25 ethyl acetate-hexane to afford 13.8 mg (100%) of the title compound as a white solid; I H NMR (CDC13) 6 2.83 1H, J 14 Hz), 2.39 III, J 14 Hz); Mass Spectrum (El) m/e 813 (M+Na).

WO 97/16182 WO 9716182PCTIUS96/1 7482 77 EXAMPLE 9A 29(Z)-Methoxycarbonyl-4,6,7,15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- 1 8-hydroxy-22-methoxycarbonyl [6x,7x, 150f, 16P3,21 f,221]D:A-Friedo- A-homo-27,30-dinor-24-oxaoleana- 1 .20(29)-dien-3 -one 00 2 0H 3 0 H H

CO

2

CH

3 Ac 0 0 OAc AcO A solution of 5.9 mg (0.028 mmol) of methyl diethlyphosphonoacetate in 1 mE of THF at 0 0 C was added potassium bis(trimethylsilyl)amide (0.O28mmol) and the solution was stirred for 30 minutes. Then it was cooled to -78'C and a solution of 4,6,7,15,16- Pentakis(acetyloxy)-2 1,22-epoxy- I 8-hydroxy-22-methoxycarbonyl- [6x,7cx, 1513,1613,21 1,221]D:A-Friedo-A-homo-27,30-dinor-24.

oxaoleana-1I-en-,3,20-dione (18.5 mg, 0.023 mmol) in 1.0 mL of THF was added. The solution was allowed to warm to 25*C for 36 h. The solution was diluted with CH2CI2 and was filtered through celite. Upon evaporation of solvent, the residue was purified by chromatography on silica gel using 50% ethyl acetate-hexane to afford 4.0 mg of the Z isomer and 5.4 mg of the E isomer (34 I H NMR (CDCl3) 6 6.00 I1H, J 1.8 Hz), 3.81 3 Mass Spectrum (El) m/e 869 (M+Na).

The following Examples 9-13 are isomeric mixtures which were prepared by the procedures described in Example 9A.

WO 97/16182 WO 9716182PCTIUS96/1 7482 78 EXAMPLE 9B 29(E)-Methoxycarbonyl-4,6,7,15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- 1 8-hydroxy-22-methoxycarbonyl [6a,7(x, 1 5,1 6f,21 P,220]D:A-Friedo- A-homo-27 .30-dinor-24-oxaoleana- 1 .20(29')-dien-3 -one MeO 2

C-

OAc I H NMR (CDC13) 6 6.30 I H, J 2.1 Hz), 3.74 I Mass Spectrum (El) m/e 869 (M+Na).

EXAMPLE 1OA 29(Z)-t-Butoxycarbonyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- 1 S-hydroxy-22-methoxycarbonyl[6u,7x,15j3,16P,2 1 f,22f]D:A-Friedo- A-homo-27..30-dinor-24-oxaoleana- 1.20(29')-dien-3 -one 0 C0 2

CH

3 WO 97/16182 WO 9716182PCTIUS96/1 7482 79 1 H NMR (CDC13) 865.92 I H, J 1.7 Hz), 1.51 9 Mass Spectrum (El) mle 911 (M+Na).

EXAMPLE IlOB 29(E)-t-Butoxycarbonyl 15,1 6-pentaki s(acetyloxy)-2 l, 2 2 -epoxy- 1 8-hydroxy-22-methoxycarbonyl[6Q,7(x, 15f,1 6j,2 1 f,22f3]D:A-Friedo- A-homo-27,30-dinor-24-oxaoleana-. 1 ,20(29)-dien-3-one G, Ac GAc I H NMR (CDC13) 5 6.24 I1H, J 2.0 Hz), 1.47 9 Mass Spectrum (El) m/e 911 (M+Na).

WO 97/16182 WO 9716182PCTIUS96/17482 80 EXAMPLE 1 I A 29(Z)-Ethoxycarbonyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1,22-epoxy-i 18hydroxy-22-methoxycarbonyl[6uc,7Qc 1 5f,1 6f,2 1 1,221]D:A-Friedo-Ahomo-27,30-dinor-24-oxaoleana- 1, 2 0(29)-dien-3-one

CO

2 Et C0 2

CH

3 Ac AcO I H NMR (CDCI3) 565.99 I1H, J =1.9 Hz), 1.26 3 H, J 7.2 Hz); Mass Spectrum (El) m/e 883 (M+Na).

EXAMPLE I11 B 29(E)-Ethoxycarbonyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 l,22-epoxy- 18hydroxy-22-methoxycarbonyl[6Q,7x, 1 5P~,1 6P3,21 1,22P]D:A-Friedo-Ahomo-27,30-dinor-24-oxaoleana- I 2 0(29)-dien-3-one '0 -C0 2

CH

3 OAc AcO WO 97/1 6182 CUS6'78 PCTIUS96/17482 81 I H NMR (CDCI3) 6 6.29 1H, J 1.9 Hz), 1.28 3 H, J 7.2 Hz); Mass Spectrum (El) m/e 883 (M+Na).

EXAMPLE 12A 29(Z)-Cyano-4,6,7,15,1I6-pentakis(acetyloxy)-2 l,22-epoxy-1I 8-hydroxy- 22-methoxycarbony[6cx,7ot, 1 5P3, 16P,21 ,22P]D :A-Friedo-A-homo- 2 7,30-dinor-24-oxaoleana- 1 2 0(29)-dien-3-one

OCN

0 C0 2

CH

3 OAc OAc 1 H NMR (CDCI3) 8 5.53 I1H, J 2.0 Hz); Mass Spectrum (APCI) m/e 831 (M+NH4).

WO 97/16182 WO 9716182PCT/US96/17482 82 EXAMPLE 12B 29(E)-Cyano-4,6,7,15,1I6-pentakis(acetyloxy)-2 1,22-epoxy- I 8-hydroxy- 22-methoxycarbony[6,7ix, 1 5J,16f0,2 1 ,223]D :A-Friedo-A-homo- 27,30-dinor-24-oxaoleana- 1 .20(29)-dien-3-one C0 2

CH

3 .,OAc GAc I H NMR (CDCJ3) 8 5.82 I1H, J 1.9 Hz); Mass Spectrum (APCI) m/e 831 (M+N114).

EXAMPLE 1 3A 29(E) -Phenyl 15,1 6-pentaki s(acetyl oxy) -2 1,22 -epoxy- I 8-hydroxy- 22-methoxycarbonyI[6a,7a, 1 5P, 1 6f,21 P,221]D:A-Friedo-A-homo- 27 3 0-dinor-24-oxaoleana- I ,20(29)-dien-3 -one

.CO

2

CH

3 GAc AcO" I H NMR (CDCI3) 8 7.37 2H, J 7 Hz), 7.31 I H, J 7 Hz), 7.27 2H, J 7 Hz), 7.05(s, I1H); Mass Spectrum (APCI) m/e 882 (M+NH-4).

WO 97/16182 WO 9716182PCTIUS96/ 17482 83 EXAMPLE 1 3B 29(Z)-phenyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- I 8-hydroxy- 22-methoxycarbonyl[6cc,7Q, 1$0, 1 6f21 f,22J]D :A-Friedo-A-homo- 27 .30-dinor-24-oxaoleana- 1 .20(29')-dien-3 -one OAc AcO I H NMR (CDC13) 8 7.46 2H, J 8 Hz), 7.35(t, 1 H, J 7 Hz), 7.33 2H, J 8 Hz), 6.80 I1H); Mass Spectrum (APCI) m/e 882 (M+N114).

M

WO 97/16182 WO 9716182PCT/US96/1 7482 84 EXAMPLE 14 4-(2-Bromobenzoyl)oxy-6,7, 15,1 6-tetrakis(acetyloxy)-2 1,22-epoxy-I 18hydroxy-22-methoxycarbonyl [6ot,7(x, 1 5fp,16f3,2Oox,2 1 ,22p]D :A- Friedo-A-homo-27 .30-dinor-24-oxaoleana- 1 -en-3-one -C0 2

CH

3 OAc Step A: 6,7,15,1 6-Tetrakis(acetyloxy)-21I,22-epoxy-4, 18dihydroxy-22-methoxycarbonyl [6uc,7a, 15Sf, 1 6f,2 1 f,22f]D:A-Friedo-A-homo-27,30.

dinor-24-oxaoleana- 1 .20(29')-dien-3 -one 0 Li CO 2

CH

3 OAc A solution of 102.1 mg (0.130 mmnole) of 4,6,7,15,16pentakis(acetyloxy)-2 l, 2 2 -epoxy- I 8-hydroxy-22-methoxycarbonyl r6(x,7cx, 15pf, 1 6P3,21 f, 2 2f3]D:A-Friedo-A-homo-27,30-dinor-24- WO 97/16182 PCT/US96/17482 85 oxaoleana- 1,20(29)-dien-3-one in 4 mL of tetrahydrofuran and 2 mL of 3M aqueous HCI was heated at 40*C for 24h. The solution was diluted with dichloromethane and the layers were separated. The organic layer was washed with 0.1 M phosphate buffer (pH then was dried over MgSO4 and concentrated. The residue was purified by silica gel chromatography with 2:1 ethyl acetate-hexane to afford 44.9 mg of the title compound as a white solid 1 H NMR (CDCl3) 8 4.20 1H, J 4.3 Hz, C4-H); Mass Spectrum (APCI): m/e 764 (M+NH4) Step B: 4-(2-Bromobenzoyl)oxy-6,7,15,16-tetrakis(acetyloxy)- 21,22-epoxy-1 8 -hydroxy-22-methoxycarbonyl- [6a,7a,150,160,210 ,22P]D:A-Friedo-A-homo-27,30dinor-24-oxaoleana- 1.20(29)-dien-3-one

O

CO

2

CH

3 OAc To a solution of 17.5 mg (23.5 gmole) of 6,7,15,16tetrakis(acetyloxy)-21,22-epoxy-4,18-dihydroxy-22-methoxycarbonyl- [6a,7a, 15P,160,21 P,22P]D:A-Friedo-A-homo-27,30-dinor-24oxaoleana-1,20(29)-dien-3-one in 0.5 mL pyridine was added 27.5 mL (237 tmole) of 2-bromobenzoyl chloride. The solution was stirred at room temperature for 4 h, then was concentrated under reduced pressure. The residue was first filtered through a plug of silica gel and then purified by HPLC (Waters RCM, t Porosil, 10 mm X 10 cm) using a mixture of 9.6:6 (5:4:1 hexane-methyl tert-butyl ether- WO 97/16182 PCT/US96/17482 -86acetonitrile:hexane) to afford 17.3 mg of the title compound as a white solid; IH NMR 6 5.67 (1H, C4-H), 7.40-7.43 (min, 2H), 7.72 (dd, 1H, J 2.2, 6.9 Hz), 7.78 (dd, 1H, J 2.3, 6.9 Hz); Mass Spectrum (APCI): m/e 946, 948 7 9 Br-M+NH4, 8 1 Br-M+NH4) Step C: 4-(2-Bromobenzoyl)oxy-6,7,15,1 6-tetrakis(acetyloxy)- 2 1,22-epoxy- I 8 -hydroxy-22-methoxycarbonyl- [6a,7a, 150,160,20u,21 P,22P]D:A-Friedo-A-homo-27,30dinor-24-oxaoleana- I -en-3-one Br Ao A solution of 0.100 g (0.108 pmole) of 4-(2bromobenzoyl)oxy-6,7,15,1 6-tetrakis(acetyloxy)-2 1,22-epoxy-1 8hydroxy-22-methoxycarbonyl[6a,7a, 15(,16(,21 f,22P]D:A-Friedo-Ahomo- 2 7 3 0-dinor-24-oxaoleana- 1, 2 0(29)-dien-3-one in 20ml of dry THF was degassed under reduced pressure and saturated with nitrogen, the procedure being repeated several times. Then 75 mg of Wilkinson's catalyst (PPh3)3RhC] were added and the solution was degassed and saturated with hydrogen in the previously described manner. The reaction vessel was then pressurized with H2 to 50 psi (3.5 atm) and shaken for 72h at 25 0 C. After that time the solvent was removed under reduced pressure. The residue was dissolved in a small amount of ethyl acetate/hexanes (ca. 1 mL) and filtered through 3 0g of silica gel eluting with 500mi of ethyl acetate hexanes The first fractions, WO 97/16182 WO 9716182PCT/US96/1 7482 87 containing the Wilkinson-catalyst (approx. 5OmL) were discarded. The fractions containing the product were combined. After removing the solvent under reduced pressure the crude product was dried in vacuo and purified by HPLC to afford 71.2 mg (71 of the title compound as a white solid; I H NMR (CDCI3) 5 1.5 3H, J 8.6 Hz, C29-H), 2.4 (in, I1H, C20-H); 5.67 (1 H, C4-H), 7.40-7.43 (in, 2H), 7.72 (dd, IlH, J 2.2, 6.9 Hz), 7.78 (dd, I1H, J 6.9 Hz); Mass Spectrum (APCI) m/e 948, 950 7 9 Br-M+N-4, 8 1 Br-M+NH4).

EXAMPLE 4,6,7,15,1 6-Pentakis(acetyloxy)-2 l, 2 2-epoxy- I 8-hydroxy-22methoxycarbonyl [6cc,7Q, 1 5P3, 1 6j3,2 1 ,22f3 ID:A-Friedo-A-homo-.27,30dinor-24-oxaoleana-2002).en..3 -one O ~1 24 621 157 3 2 5 O As described in Scheme 1, 4 ,5, 6 ,l,l 6 -pentakis(acetyloxy)- 2 l,22-epoxy- I 8 -hydroxy-22..methoxycarbonyl[6a ,7u, 1 5P3, 1 6f,21 P1, 220]D :A-Freido-A-homo..27,30.dinor..24-oxaoleana- 1 ,20(29)-diene-3one, isolated from Spachea correa in liquid ammonia with lithium metal will result in the reduction of the ClI olefin group to produce the saturated lactone.

WO 97/16182 PCT/US96/17482 88 EXAMPLE 16 4,6,7,15,16-Pentakis(acetyloxy)-21,22-epoxy-18-hydroxy-22methoxycarbonyl[6a,7a,150,1 6,20a,2 1,22p]D:A-Friedo-A-homo- 27,30-dinor-24-oxaoleana-3-one

O

H OH CO2CH3 I f I OAc O OAc SOAc AcO

A

A solution of 50 mg of 4 6 7 ,15,16-pentakis(acetyloxy)- 21,22-epoxy-1 8 -hydroxy-22-methoxycarbonyl [6a,7a,150,16p,213,- 223p]-D:A-Friedo-A-homo-27,30-dinor-24-oxaoleana-20(29)-en-3-one in 50ml of dry THF is degassed under reduced pressure and saturated with nitrogen, the procedure being repeated several times. 25mg of Wilkinson's catalyst, (PPh3)3RhC, are added and the solution is degassed and saturated with hydrogen in the previously described manner. The reaction vessel is then pressurized with H2 to 15 psi (latm) and shaken for 65h at 25 0 C. After that time the solvent is removed under reduced pressure. The residue is dissolved in a small amount of ethyl acetate/hexanes (ca. 1 mL) and filtered through of silica gel eluting with 500 ml of ethyl acetate/hexanes The first fractions, containing the Wilkinson-catalyst (approx. 50mL) are discarded. The fractions containing the product are combined. After removing the solvent under reduced pressure the crude product is dried in vacuo and purified by HPLC to produce the title compound.

I

WO 97/16182 PCT/US96/17482 89- EXAMPLE 17 4,6,7,15,16-Pentakis(acetyloxy)-21,22-epoxy-18-hydroxy-22methoxycarbonyl[6a,7a, 1513,163,21 P,22p]D:A-Friedo-A-homo-27,30dinor-24-oxaoleane

O

H OH C2CH OAc OAc 0 OAC AcO Ac A solution of 213 mg of 4 6 7 ,15,1 6 -pentakis(acetyloxy)- 2 1,22-epoxy-18-hydroxy-22-methoxycarbonyl-[6a,7(, 150, 16p,21 3,- 2 2 p]D:A-Friedo-A-homo-27,30-dinor-24-oxaoleana-3-one in 2 mL of dichloromethane is cooled to 0°C. Then 0.68 mL of a 1.02M solution of lithium tri-tertbutoxyaluminum hydride is added and the solution is stirred at 0°C for 18h. The reaction is quenched with 10 mL of aqueous H2S04 and then is diluted with 50 mL of dichloromethane.

After the layers are separated, the organic phase is dried over Na2SO 4 and concentrated.

The residue is dissolved in a mixture of 5 mL of dichloromethane and 1 mL of triethylsilane. Then 0.8 mL of boron trifluorideetherate is added and the solution is stirred at room temperature. After 2 h, the reaction is quenched by addition of saturated aqueous NaHCO 3 solution and dichloromethane. The layers are separated and the organic layer is washed with and brine, saturated aqueous NaHCO 3 dried over MgSO4, and concentrated. The residue is purified by silica gel chromatography using S (hexane:t-butylmethylether:acetonitrile 8:4:1) to produce the title compound.

WO 97/16182 WO 9716182PCT/US96/17482 90 EXAMPLE 18 4,6,7,15,1 6-Pentakis(acetyloxy)-20(29),2 1,22-diepoxy- 1 8-hydroxy-22methoxycarbonyl [6x,7(x, 1 S3, 1 6J,2 1 j,22J3]D:A-Friedo-A-homo-27,30..

dinor-24-oxaoleana-3 -one AcO~- A solution of 50 mg of 4 6 7 ,l5,16-pentakis(acetyloxy)- 21,22-epoxy- I 8 -hydroxy-22-methoxycarbonyl 6ix,7Qx, 15 0f,16 6,2 1 P, 223] D: A-Friedo-A-homo-27 3 O-dinor-24-oxaoleana-20(29) -en-3 -one and 150 mg of m-chloro perbenzoic acid (80 85%; remainder water; 0.7 mmole) in 5mi of CHC1 3 is stored at OTC for 21 days. The solution is diluted with 50 ml of dichloromethane, 20 ml of a saturated aqueous solution of NaHCO 3 is added and the layers are separated. The organic layer is consecutively washed with 20 mL of I M hydrochloric acid and saturated aqueous sodium chloride then is dried over MgSO 4 and concentrated. The residue is purified by silica gel chromatography with 1: 1 ethyl acetate-hexane to produce the title compound.

WO 97/16182 WO 9716182PCTIUS96/1 7482 91 EXAMPLE 19 4,6,7,15,16 -Pentaki s(acetyl oxy) -2 1,22 -epoxy- I 8-hydroxy-22-methoxycarbonyl[6ux,7(x, 1 5P,.1 6P3,21 f,22131D:A-Friedo-A-homo-27 ,30-dinor-24oxaoleana-3.20-dione AcO~N.c A solution of 13.7 mg of 4 6 ,7,l5,16-Pentakis(acetyloxy)- 21,22-epoxy-I 8 -hydroxy-22-methoxycarbony..[6,7., 151,1 6f,21 2213] A-Friedo-A-homo -27 3 O0-dinor-24 -oxao leana-20(29)-en- 3-one in 12 ml of 1, CH2C1 2

/CH

3 OH) is cooled to -78'C and 03 is bubbled into the solution until it contained a blue color. The solution is then purged with nitrogen for 3 minutes and 3 drops of Me2S is added. The solution is allowed to warm to 25'C for 14 hours. Volitiles are removed by vacuum and the residue is purified by chromatography on silica gel using 25 ethyl acetate-hexane to produce the title compound.

WO 97/16182 WO 9716182PCT/US96/1 7482 92 EXAMPLE 29(Z)-Methoxycarbonyl-4,6,7,15, 16-pentakis(acetyloxy)-2 l,22-epoxy- I 8-hydroxy-22-methoxycarbonyl [6u,7ux, 1 5p~,1 6fP,2 1 ,22P]D :A-Friedo- A-homo-27,30-dinor-24-oxaoleana-20(29)-en-3 -one

.CO

2 C H 3

CO

2

CH

3 GAc AcO

,P~

EXAMPLE 29(E)-Methoxycarbonyl-4,6,7,15,1 6-pentakis(acetyloxy)-2 1,22-epoxy- 1 8-hydroxy-22-methoxycarbonyl [6ux,7, 1 5p, 1 6fP,2 1 f,22P]D: A-Friedo- A-homo-27..30-dinor-24-oxaoleana-20(29)-en-3 -one '0 0C0 2 0H 3 A solution of 5.9 mg of methyl diethlyphosphonoacetate in I mL of THF at 0 0 C is added potassium bis(trimethylsilyl)amide (0.028 mmol) and the solution is stirred for 30 minutes. Then it is cooled to WO 97/16182 WO 97/ 6182PCT/US96/1 7482 93 -78*C and a solution of 4 6 7 lSl 6 -pentakis(acetyloxy)-21,22..epoxy- 1 8 -hydroxy-22-methoxycarbonyl[6Qx,7ac, 150,1 6P,2 1 P,22f3]D:A-Friedo- A-homo-27,30-dinor-24-oxaoleana-32O..dione (18.5 mg), in 1.0 mL of TI-F is added. The solution is allowed to warm to 25'C for 36 h. The solution is diluted with CH2CI2 and is filtered through celite. Upon evaporation of solvent, the residue is purified by chromatography on silica gel using 50% ethyl acetate-hexane to separate the Z isomer and the E isomer.

The following Examples, Examples 21A-24B, are prepared by the procedures described in Examples 20A and EXAMPLE 21 A 2 9(Z)-t-Butoxycarbonyl-4 15,1 6-pentakis(acetyloxy)-21I,22-epoxy- 1 8 -hydroxy-22-methoxycarbonyl [6c,71x, 15 f,1 6f,2 1 J,22f0D:A-Friedo- A-homo-27 3 O-dinor-24-oxaoleana-20(29)-en-.3 -one

CO

2 tBu 0 60 2

CH

3 OAc WO 97/16182 WO 9716182PCT/US96/17482 94 EXAMPLE 21B 29(E) -t-B utoxycarbonyl 15,1 6 -pentakis (acetyloxy-2 1 2 2 -epoxy- I 8 -hydroxy-22-methoxycarbonyl [6ac,7c(, 1 50,16f3,2 10, 22 1]D:A-Friedo.

A-homo- 2 7 3 O-dinor-24-oxaoleanJa2(29)en3 -one tBUO 2CI 0 H o C 02 CH3 OAc QAc EXAMPLE 22A 29(Z)-Ethoxycarbonyl46,715,1 6 -pentakis(acetyloxy)-2l, 2 2-epoxy- 18hydroxy-22-methoxycarbonyl[6(x,7ax I 5j3,1I60,210f,22131D:A-Friedo-A.

homo- 2 7 3 O-dinor.24xaoeana20(29)en-3 -one

CO

2 Et 0 H 00 2

CH

3 GAc WO 97/16182 PCT/1JS96/1 7482 95 EXAMPLE 22B 29(E)-Ethoxycarbonyl-4,6,7, 15,1 6 -pentakis(acetyloxy)-2 1, 2 2 -epoxy- 18hydroxy-22-methoxycarbony1[6,7Qx, 1 5f0,1 6f,2101,22f]D:A-Friedo-Ahomo-27 3 O-dinor-24-oxaoleana-20(29y..en-3 -one EtO 2

C'

0C0 2

CH

3 OAc AcO EXAMPLE 23A 29(Z)-Cyano-4,6,7, 15,1 6 -pentakis(acetyloxy)-2 l, 2 2-epoxy- I 8-hydroxy- 22-methoxycarbonyl[6u,7a, 15p3,1 61,210J,22f]D:A-Friedo-A-homo- 27 3 O-dinor- 2 4 -ox aoleana-20(29)en-3.-one

,CN

C0 2

CH

3 QAc WO 97/16182 PCTIUS96/1 7482 96 EXAMPLE 23B 29(E)-Cyano-4,6,7, 15,1 6 -pentakis(acetyloxy)-2 1, 2 2-epoxy- I 8-hydroxy- 2 2 -methoxycarbony[6Q,7cx, 5j, 1 60,2 1 ,22p] D:A-Friedo-A-homo- .27 3 -dinor-24-oxaoleana-20(29)-en-3 -one C0 2

CH

3 GAc EXAMPLE 24A 29(E)-Phenyl-4,6,7, 15,1 6 -pentakis(acetyloxy)-2 1,22-epoxy- I 8-hydroxy- 22-methoxycarbony[6x,7(x, 1 5f,1 6f,21 J,22t0]D:A-Friedo-A-homo-.

2 7 3 O-dinor-24-oxaoleana20(29..en-3.-one OAc AcO" WO 97/16182PCUS6178 PCT/US96/17482 97 EXAMPLE 24B 29(Z)-Phenyl-4,6,7, 15,1 6-pentakis(acetyloxy)-2 1,22-epoxy-I 8-hydroxy- 22-methoxycarbonyl[6a,7x, 15fP,1 6f,21 P3,22P]D :A-Friedo-A-homo- 2 7 3 -dinor- 2 4-oxaoleana-20(29)-en-3-one OAc EXAMPLE 4 Bromobenzoyl oxy)-6,7, 15,1 6-tetrakis (acety lox y) -21,22-epoxy-i 18hydroxy-22-methoxycarbonyl[6a,7u, 150f,1I6f3,21 1,22f0D:A-Friedo-Ahomo-27..30-dinor-24-oxaoleana-3 -one OAc

OAC

WO 97/16182 WO 9716182PCTIUS96/17482 98 Step A: 6,7,15,1 6-Tetrakis(acetyloxy)-2 1,22-epoxy-4, 18dihydroxy-22-methoxycarbonyl[6,7x, 15 p, 1 6P3,21 fP,223] D :A-Friedo-A-homo-27,30-dinor-24-oxaoleana-20(29)-en- 3-one o A c

H

A solution of 102.1 mg of 4,6,7,15,16-pentakis (acetyloxy)-2 1,22-epoxy- I 8-hydroxy-22-methoxycarbonyl- [6(x,7ax, 15Pf, 1 6f,21 f,22f]D:A-Friedo-A-homo-27,30-dinor-24oxaoleana-20(29)-en-3 -one in 4 mL of tetrahydrofuran and 2 mL of 3M aqueous HCl is heated at 40TC for 24h. The solution is diluted with dichioromethane and the layers are separated. The organic layer is washed with 0. 1 M phosphate buffer (pH then is dried over MgSO4 and concentrated. The residue is purified by silica gel chromatography with 2:1 ethyl acetate-hexane to produce the title compound.

Step B: 4- Bromobenzoy lox y)-6,7,1 15,16 -tetraki s(acety loxy) 21,22-epoxy-i I -hydroxy-22-methoxycarbonyl- [6(x,7cx, 1,1 6j,21 f,22f3JD:A-Friedo-A-homo-27,30dinor-24-oxaoleana-20(29)-en-3 -one WO 97/16182 WO 9716182PCT/US96/17482 99 0 OH

CO

2

CH

3 0 OAc o ',,OAc OAc Br0 0 To a solution of 17.5 mg of 6,7,15,16-tetrakis(acetyloxy)- 21 ,22-epoxy-4, 18-dihydroxy-22-methoxycarbonyl 1 5P3,1 6P3,21(P, 22P3] D: A-Friedo-A-homo-27,30-dinor-24-oxaoleana-20(29)-en-3 -one in 0.5 mL pyridine is added 27.5 [tL (237 jimole) of benzoyl chloride.

The solution is stirred at room temperature for 4 h, then is concentrated under reduced pressure. The residue is first filtered through a plug of silica gel and then purified by HPLC (Waters RCM, Rt Porosil, 10 mm. X cm) using a mixture of 9.6:6 (5:4:1 hexane-methyl tert-butyl etheracetonitrile:hexane) to produce the title compound.

Step C: 4- (2 -B romobenzoy lox y)-6,7,15,16 -tetrakis (acety lox y) 21,22-epoxy- I 8-hydroxy-22-methoxycarbonyl- [6Qx,7Q, 1513,1613,21 1,221]D:A-Friedo-A-homo-27,30dinor-24-ox aoleana- 3-one WO 97/16182 PCT/US96/17482 100- H OH CO2CH3 0 OAc S OAc Br O O A c OAc Br 0 A solution of 0.100 g of 4-(2-bromobenzoyloxy)-6,7,15,16tetrakis(acetyloxy)-21, 2 2-epoxy-18-hydroxy-22-methoxycarbonyl- [6a,7a, 153,16,21 ,22P]D:A-Friedo-A-homo-27,30-dinor-24oxaoleana-20(29)-en-3-one in 20ml of dry THF is degassed under reduced pressure and saturated with nitrogen, the procedure being repeated several times. Then 75 mg of Wilkinson's catalyst (PPh3)3RhCl are added and the solution is degassed and saturated with hydrogen in the previously described manner. The reaction vessel is then pressurized with H2 to 50 psi (3.5 atm) and shaken for 72h at 25 0

C.

After that time the solvent is removed under reduced pressure. The residue is dissolved in a small amount of ethyl acetate/hexanes (ca.

1 mL) and is filtered through 30g of silica gel eluting with 500ml of ethyl acetate hexanes The first fractions, containing the Wilkinson-catalyst (approx. 50mL) are discarded. The fractions containing the product are combined. After removing the solvent under reduced pressure the crude product is dried in vacuo and purified by HPLC to produce the title compound.

Claims

1. A compound of structural Formula I: R29 R29b R R 12 H 11 2 1 H9 or a pharmaceutically acceptable salt, crystal form, or hydrate, wherein: X is: O, S, NH or H and R 1 a is: a single bond, or a double bond when R 4 is absent; b and c are independently: a single bond or a double bond; n is: 1 to 4; m is: 1 to 4; ris: Oor 1; sis: 0 or 1; R 1 is: a) -To 7)*4 r"J \~i ouj H, or (C 1 -C 6 )-alkyl, wherein alkyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1 -C 6 )-alkoxy, vinyl, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 alkyl, CO 2 C 1 -C 6 -alkyl, CONR 1 aR 2 NRiaR 2 NR 1 aCOC 1 -C 6 -alkyl, aryl, wherein aryl is defined as phenyl or naphthyl, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1 -C 6 )-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyl, C0 2 C 1 -C 6 -alkyl, CONRiaR2, NRlaR2, NR la COC1- C 6 -alkyl and any two of adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, heteroaryl, wherein heteroaryl is defined as a 5 or

6-membered ring substituted with one and two heteroatoms selected from O, S, N, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1 -C 6 )-alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyl, CO 2 C 1 -C 6 -alkyl, CONRlaR 2 NRlaR 2 NRlaCOC 1 -C6-alkyl, any two adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 IN:\LIBAA]01587.doc:TAB 102 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring: Ria and R 2 are independently: a) (Cl-0 6 )-alkyl, or b) aryl, wherein aryl is phenyl or naphthyl; R 3 is: a) -(C 1 -C 6 )-alkyl, alkyl as defined above; b) -(Cl-C 6 )-alkenyl, wherein alkenyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, 00 2 H, 000 1 -C 6 -alkyl, C0 2 CI-C 6 -alkyl, CONRlaR2, NRlaR2, NRlaCOC, -C 6 -alkyl, aryl as defined above, and heteroaryl as defined above; C) -(Cl-C 6 )-alkynyl, wherein alkynyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COCI-C 6 -alkyl, C0 2 C 1 -C 6 -alkyl, CONRlaR2, NRlaR2, NRlaCOC1-C6ealkyl, aryl as defined above, and heteroaryl as defined above, d) -aryl, aryl as defined above, or e) -heteroaryl, heteroaryl as defined above; R 4 is: a) absent and a is a double bond; 00 0..000 0S 0 seee 0005 ROOm 5 0 bo S. S ass. See. 5 S 05 0@ S 00 @5 0 S S Seeg 0@50 ~0e.. @0500e 0 b) -H, c) -OH, d) =O, e) -O[(C=O)Or]sC 1 -C 1 0 -alkyl, alkyl as defined above, f) -O[(C=O)Or1,C2-C 1 0-alkenyl, as defined above, g) -o[(C=O)Or]sC 2 -C 6 -alkynyl, alkynyl as defined above, i) -O[(C=O)O]ryl, arC-yla fiedabve j) O0[(C=O)O~]roaryl, traryl as defined above, k) -O[(CH2)nO(CHheteroaryl, heteroaryl as defined above, k) -O(CH 2 )nO(CH 2 )mhraryl, traryl as defined above, m) -OC(=O)NRlaR 2 n) -0S0 2 R 3 or o) -NRlaR2; R 29 a and R29b are independently: a) -H, b) -(CH2)s-O[(C=O)OrsC-C 10 .alkyl, alkyl as defined above, c) -(CH2)s-O[(C=O)OrJsC 2 -C 1 0 alkenyl, alkenyl as defined abc d) -(CH2)s-O[(CO)O]sC 2 .C 6 alkynyl, alkynyl as defined abov e) -(CH2)s-O[(C=O)Or]s(C 3 C 7 )-cycloalkyl, 'ye, e, C~O L~ r *J s- L4f J fN:\LIBAA]O1

587.doc:TAB M 103 g) -(CH2),-O[(C=0)Orlsheteroaryl, heteroaryl as defined above, h) 0 H2)s-O(CH 2 )flO(CH 2 )mheteroalyl heteroaryl as defined above, i) -(CH2)s0(CH2)nO(CH 2 )maryl, aryl as defined above, j) -(CH 2 ),-0C(=0)NRlaR2, k) -(CH 2 )S-0S0 2 R 3 1) -(0 1 -C 6 )-alkyl, alkyl as defined above, or M) -(0 2 -0 6 )-alkenyl, alkenyl as defined above; R 2 9a and R29b together are =0 or =C(Cl-Cl 0 -alkyl) 2 alkyl being as defined above. 2. The compound of structural Formula 1, as recited in claim 1, 29a 29b C0 2 CH 3 'QAc 'OAc @0 0@00* 05 5 05 5 5 S S. SS @000 .0050 0.00. 0005 50 005 5 5 C See.. 0~00 S. 15 5 00S 000 0505 @SSe 5 05 05 5 @5.e 20 0550 5 5 Se S. 0 05 55 5 5 0 *SS. *.SS 25 0 or a pharmaceutically acceptable salt, crystal form, or hydrate, wherein: X is: 0, S, orNH; a is: a single bond: b and c are independently: n is: m is: r is: S is: R 1 is: a) I-X a single bond or a double bond; 1 to 4; 1 to 4; 0Oorl1; o or 1; H, or (01-C6)-alkyl, wherein alkyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, vinyl, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, C001-C6- alkyl, 00201 -C 6 -alkyl, CONR 1 aR2, NR 1 aR2, NR 1 aCOCI C 6 -alkyl, aryl, wherein aryl is defined as phenyl or naphthyl, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COCI-C 6 -alkyl, 00201 -C 6 -alkyl, CONR 1 aR2, NR 1 aR2, NR 1 aCOC 1 C 6 -alkyl and any two of adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, heteroaryl, wherein heteroaryl is defined as a 5 or fN:\LIBAAJO1 587.doc:TAB 104 6-membered ring substituted with one and two heteroatoms selected from 0, S, N, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyi, C0 2 C 1 -C 6 -alkyl, CON R 1 aR2, NRlaR2, NRlaCOC 1 -C 6 -alkyl, any two adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring: Rla and R 2 are independently: a) (Cl-C 6 )-alkyl, or b) aryl, wherein aryl is phenyl or naphthyl; R 3 is: a) -(C-C 6 )-alkyl, alkyl as defined above; b) -(Cl-C 6 )-alkenyl, wherein alkenyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, 1, (Cl-C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyl, C0 2 C 1 -C 6 -alkyl, CONR 1 aR2, NR 1 aR2, NR 1 aCOCi C 6 -alkyl, aryl as defined above, and heteroaryl as defined above; C) -(Cl-C 6 )-alkynyl, wherein alkynyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, 20 (C 1 -C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyl, C0 2 C 1 -C 6 -alkyl, CON RI R 2 NR 1 aR2, NR 1 aCOCl..c 6 alkyl, aryl as defined above, and heteroaryl as defined above, d) -aryl, aryl as defined above, or e) -heteroaryl, heteroaryl as defined above; R 4 is: a) absent and a is a double bond; *000 I 5.6. 6@ I SS S S *e ge 0e 0eOOe SO... *500 5* S 0@ S* C S S S S.. OSSb 0 S be 0* S 555e 0 S 0 0 O @5 0 .5 0O I S 0 **s5 I- -H, -OH, =0, -O[(C=O)Or]sC 1 -CI 0 -alkyl, alkyl as defined above, 5 C 2 C 1 0 -alkeny, as defined above, -O[(C=O)Or]sC 2 -C 6 alkynyl, alkynyl as defined above, -O[(C=O)Orls(C 3 -C 7 )-cycloalkyl, -O[(C=O)Orsaryl, aryl as defined above, -O[(C=O)Or],heteroaryl, heteroaryl as defined above, -O(CH2)flO(CH 2 )mheteroaryl, heteroaryl as defined above, -O(CH2)nO(CH 2 )maiy, aryl as defined above, -OC(=O)NRl aR2, -0S0 2 R 3 or -NR 1 aR2, -v A L' &4 [N:\LIBAAO1 587.doc:TAB R29a and R29b are independently: a) -H, b) -(CH2)s-O(C0O)OrlIsClO-alkyl, alkyl as defined above, c) -(CH2)s-O[(C=O)Or~sC 2 -Clo-alkenyl, alkenyl as defined above, d) -(CH2)s-O(C=O)Or~sC 2 -C 6 -alkynyl, alkynyl as defined above, e) -(CH2)s0[I(C=O)Orls(C 3 -C 7 )-cycloalkyl, f) -(CH2)s-O[(C=O)Orsaryl, aryl as defined above, g) -(CH2)s-O[(C=O)Orlsheteroaryl, heteroaryl as defined above, h) -(CH2)s-O(CH 2 )flO(0H 2 )mheteroaryl, heteroaryi as defined above, i) -(CH2)s-O(CH 2 )flO(CH 2 )maryl, aryl as defined above, j) -(CH 2 ),-OC(=O)NRlaR 2 k) -(OH 2 )s-OSO 2 R 3 1) -(C 1 -C 6 )-alkyl, alkyl as defined above, or M) -(C 2 -C 6 )-alkenyl, alkenyl as defined above; R 2 9a and R29b together are =0 or =C(Cl-C 10 -alkyl) 2 alkyl being as defined above. 3. The compound of structural Formula!1, as recited in claim 2, R 2a R 2 9b 12H 19 2 00*c 1j 8 1 7 22 0 2 H9 H 14 OH16 *b 10 25 5 QAc 003. 26 0 Q 24 6 QAc OAc Ac 23"' or a pharmaceutically acceptable salt, crystal form, or hydrate, wherein: 0 Xis: 0; 20 a is: a single bond; b band care OS independently: a single bond or a double bond; S n is: i to 4; M is: 1 to 4; :25 r is: 0Oorl1; s is: 0Oor 1; R 1 is: a) H, or b) (01-06)-alkyl, wherein alkyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, vinyl, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 alkyl, C0 2 C 1 -C 6 -alkyl, CONR 1 aR2, R 1 a2NRacc-C-alkyl, aryl, [NALIBAAO1 587.doc:TAB 106 wherein aryl is defined as phenyl or naphthyl, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, 1, (Cl-0 6 )-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COO 1 -C 6 -alkyl, 00201 -C 6 -alkyl, CON R aR2, NW aR2, N R 1 aCOC 1 C 6 -alkyl and any two of adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, heteroaryl, wherein heteroaryl is defined as a 5 or 6-membered ring substituted with one and two heteroatoms selected from 0, S, N, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, C0C1 -C 6 -alkyl, 00201 -C 6 -alkyl, CON R 1 aR2, NRlaR 2 NRlaCOc 1 -c 6 -alkyl, any two adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring: Ria and R 2 are independently: a) (C 1 -0 6 )-alkyl, or b) aryl, wherein aryl is phenyl or naphthyl, R 3 is: a) -(C 1 -C 6 )-alkyl, alkyl as defined above, b) -rlay s eie above, o -arye, aryhtay as defined or ve R 4 is: a) -O[(C=O)Or]sCi-Cl 0 -alky, alkyl as defined above, b) -O[(C=O)Or]s(C3-C 7 )-cycloalkyl, c) -O[(C=O)Or] 5 aryl, aryl as defined above, d) -O[(C0)Orlsheteroaryl, heteroaryl as defined above, e) -O(CH 2 )nO(CH 2 )mheteroaryl, heteroaryl as defined above, f) -O(CH2)flO(0H 2 )maryl, aryl as defined above, -OC(=O)NRlaR 2 or h) -0S0 2 R3; 30 R 2 9a and R29b are independently: a *0 *0 C a a S *aee 0000 b) -(CH2)s-O[(C=O)Or]sC 1 -Cl 0 alkyl, alkyl as defined above, c) -(CH2)s-O[(C=O)O]sC 2 -Clo-alkeny, alkenyl as defined above, d) -(CH2)s-O[(C=O)Or]sC 2 C 6 -alkynyl, alkynyl as defined above, e) -(CH2)s-O[(C=O)Or]s(C 3 -C 7 )-cycloalkyl, f) (CH2)s-O[(C=O)O]saryl, aryl as defined above, g) -(CH2)s-O[(C=O)Or]sheteroaryl, heteroaryl as defined above, h) -(CH2)s-O(CH 2 )flO(CH 2 )mheteroaryl, heteroaryl as defined above, i) -(CH2)S-O(CH 2 )flO(CH 2 )maryl, aryl as defined above, j) -(CH 2 )s-OC(=O)NRlaR 2 IN:\LIBAA]O1 587.doc:TAB k) -(CH 2 )S-0S0 2 R 3 1) -(Cl-C 6 )-alkyl, alkyl as defined above, or m) -(C 2 -C 6 )-alkenyl, alkenyl as defined above; R 29 a and R29b together are =0 or =C(Cl-C 10 -alkyl) 2 alkyl being as defined above. 4. The compound of structural Formula 1, as recited in claim 3, or a pharmaceutically acceptable salt, crystal form or hydrate, wherein: R 4 is: .440 a 20 .0 9 *9 0 0 3 0* 00 0049 *0040 4e 00p *400 0* *00 0 0 4 Oeegg P000 00 0* a 000 099 a) -OI(C=O)Or]saryl, wherein aryl is defined as phenyl or naphthyl, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (C 1 -C 6 )-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyl, C0 2 C 1 -C 6 -alkyl, CONRlaR 2 NRlaR 2 NRlaCOCI -C 6 -alkyl and any two of adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or b) O0[(C0)Or]sheteroaryl, wherein heteroaryi is defined as a 5 or 6-membered ring substituted with one and two heteroatoms selected from 0, S, N, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, 1, (Cl-C 6 )-alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COCI-C 6 -alkyl, C0 2 C 1 -C 6 -alkyl, CONRlaR 2 NRlaR 2 NRlaC0C 1 -C 6 -alkyl, 0 0 0 #g *4 0 0000 0540 0 0 0 0 00 *0 4 S 00 0 4 0 *S0* [N:\LIBAA 101 587.doc:TAB 108- any two adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing I or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring. The compound of structural Formula 1, as recited in Claim 1, R 2 9b C 12 s 4, 00000 06 0 0 0 0 s o Goo. OAc or a pharmaceutically acceptable salt, crystal form, or hydrate, wherein: X is: a is: 00 0 0. 0 0 1 000 H and RI; a single bond; b and c are independently: a single bond or a double bond; n is: m is: r is: I to 4; 1 to 4; o or 1; o or 1; s is: R 1 is: a) H, or b) (C 1 -C 6 )-alkyl, wherein alkyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, I, (C 1 -C6)-alkoxy, vinyl, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 alkyl, C0 2 C 1 -C 6 -alkyl, CONR 1 aR2, NRiaR2, NR1aCOC 1 -C 6 -alkyl, aryl, wherein aryl is defined as phenyl or naphthyl, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1 -C 6 )-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC1-C 6 -alkyl, C0 2 C 1 -C 6 -alkyl, CONRiaR2, NRiaR2, NRiaCOC 1 see* 10 C 6 -alkyl and any two of adjacent substituents can be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, heteroaryl, wherein heteroaryl is defined as a 5 or 6-membered ring substituted with one and two heteroatoms selected from 0O, S, N, unsubstituted or substituted with one, two or three substituents **515 selected from the group consisting of: Br, CI, F, I, (C 1 -C 6 )-alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COCI-C 6 -alkyl, C0 2 C 1 -C 6 -alkyl, CONRlaR2, NRlaR 2 NRlaCOCi-C6-alkyl, any two adjacent substituents ca be joined to form a 6- or 7-membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents 2 can be joined together to form a benzo-fused ring; Rla and R 2 are independently: a) (C 1 -C 6 )-alkyl, or b) aryl, wherein aryl is phenyl or naphthyl; R3 is: a) -(C 1 -C 6 )-alkyl, alkyl being as defined above; 5 b) -(C 1 -C)-alkenyl, wherein alkenyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1 -C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyl, CO 2 C 1 -C 6 -alkyl, CONR 1 aR2, NRiaR2, NR 1 aCOC 1 -C 6 -alkyl, aryl as defined above, and heteroaryl as defined above; c) -(C 1 -C)-alkynyl, wherein alkynyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1 -C)-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyl, C0 2 C 1 -C 6 -alkyl, CONRiaR2, NRlaR 2 NRlaCOC -C6-alkyl, aryl as defined above, and heteroaryl as defined above, d) -aryl, aryl as defined above, or e) -heteroaryl, heteroaryl as defined above; R 4 is: a) absent and a is a double bond; b) -H, A4/ c) -O H -u d) =0, [N:\LIBAA]O1587.doc:TAB 110 e) 0[I(C0)Or~sCi-CiO-alkyl, alkyl as defined above, f) 0 [I( 0 %OrIsC2-COalkeny, as defined above, g) -O[(C=0)Or]sC 2 -C 6 -alkynyl, alkynyl as defined above, h) 00 )Ors(C3-C7)-cycloalkyl, -O[(C=O)Orlsaryl, aryl as defined above, j) 0 [(C=O)Or]sheteroaryl, heteroaryl as defined above, k) -O(CH2)nO(CH 2 )mheteroaryl, heteroaryl as defined above, 1) -O(CH2)nO(CH 2 )maryl, aryl as defined above, m) -OC(=O)NRlaR 2 n) -0S0 2 R 3 or -NRlaR2; R 2 9a and R29b are independently: -H, b) -(CH2)s-O[(C=O)Or~sC-ClO-alkyl, alkyl as defined above, i c) -(CH2)sO[(C0)Or]sC 2 -ClO-alkenyl, alkenyl as defined above, d) -(CH2)sO[(C0)Or~sC 2 -C 6 -alkynyl, alkynyl as defined above, e) -(CH2)s-O[(C=O)Orls(C 3 -C 7 )-cycloalkyl, -(CH2)s-O[(C=O)Or~saryl, aryl as defined above, 9) -(CH2)s-O[(C=O)OrJsheteroaryl, heteroaryl as defined above, h) -(CH2)s-O(CH 2 )flO(CH 2 )mheteroaryl, heteroaryi as defined above, -(CH2)S-O(CH2)nO(CH 2 )maryl, aryl as defined above, j) -(CH 2 )s-OC(=0)NRlaR 2 k) -(CH 2 )s-oS0 2 R 3 1) -(C-C 6 )-alkyl, alkyl as defined above, or 25 M) -(C 2 -C 6 )-alkenyl, alkenyl as defined above; R29a and R29b together are =0 or =C(Cl-0 10 -alkyl) 2 alkyl being as defined above. 6. The compound of structural Formula 1, as recited in claim e9a R29b 12H 19 2 c 1 1i 8 1 7 22 2 1 H9 ,H O 4H 1 8C2H O 24 5 6 7 OAc OAc Ac 23^ R 4 or a pharmaceutically acceptable salt, crystal form, or hydrate, wherein: C 4 Xis: H andR1 a is: a single bond; NT 0 [N:\LIBAAIO1 587.doc:TAB b and c are independently: a single bond or a double bond; n is: 1 to 4; m is: 1 to 4; ris: 0 or 1; sis: 0 or 1; R1 is: a) H, or b) (C 1 -C 6 )-alkyl, wherein alkyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1 -C 6 )-alkoxy, vinyl, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 alkyl, C0 2 C 1 -C 6 -alkyl, CONRlaR2, NRlaR 2 NRlaCOC 1 -C6-alkyl, aryl, *see wherein aryl is defined as phenyl or naphthyl, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1 -C6)-alkoxy, phenyl, phenoxy, cyano, nitro, hydroxy, CHO, 5 CO 2 H, COC1-C 6 -alkyl, C0201-C 6 -alkyl, CONR1aR2, NR 1 aR2, NR1aCOC 1 C 6 -alkyl and any two of adjacent substituents can be joined to form a 6- or 7- membered fused ring said ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, heteroaryl, wherein heteroaryl is defined as a or 6-membered ring substituted with one and two heteroatoms selected :20 from O, S, N, unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, Cl, F, I, (C 1 -C 6 )-alkoxy, cyano, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyl, C0 2 C 1 C 6 -alkyl, CONRlaR 2 NRlaR 2 NRlaCOC 1 -C6-alkyl, any two adjacent substituents can be joined to form a 6- or 7-membered fused ring said 25 ring containing 1 or 2 oxygen atoms and the remainder carbon atoms, or any two adjacent substituents can be joined together to form a benzo-fused ring: Rla and R 2 are independently: a) (C 1 -C 6 )-alkyl, or b) aryl, wherein aryl is phenyl or naphthyl; R 3 is: a) -(C 1 -C 6 )-alkyl, alkyl as defined above; b) -(C 1 -C 6 )-alkenyl, wherein alkenyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, (C 1 -C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyl, C02C1-C 6 -alkyl, CONR 1 aR2, NRiaR2, NR 1 aCOC 1 -C 6 -alkyl, aryl as defined above, and heteroaryl as defined above; c) -(C 1 -C 6 )-alkynyl, wherein alkynyl is unsubstituted or substituted with one, two or three substituents selected from the group consisting of: Br, CI, F, I, ~j p~(C 1 -C 6 )-alkoxy, cyano, oxo, nitro, hydroxy, CHO, CO 2 H, COC 1 -C 6 -alkyl, [N:\LIBAA]O1 587.doc:TAB R4 is: 00201 -C 6 -alkyl, CONR 1 aR2, NR 1 aR2, NR 1 aCoCi -C 6 -alkyl, aryl as defined above, and heteroaryl as defined above, d) -aryl, aryl as defined above, or e) -heteroaryl, heteroaryl as defined above; a) -OH, b) O0[(C=O)Or]sCi-CO-alkyl, alkyl as defined above, C) 0 00 )Ors(C307)cycloalkyl, d) -O[(C=O)Orlsaryl, aryl as defined above, e) -0[(C=O)Orlsheteroaryl, heteroaryl as defined above, f) -O(CH2)flO(0H 2 )mheteroaiyl, heteroaryl as defined above, g) -O(CH2)nO(0H 2 )maryl, aryl as defined above, h) -O0(=O)NRIaR2, or i) -0S0 2 R3; independently: a) -H, @000 0 10 0~ 0 0@ 0 0 SW S S *505 *0 0 15 R 2 9a and R29b are b) -(CH2)s-OI(C=O)Or~sC-ClO-alkyl, alkyl as defined above, c) -(CH2)s-O[(C=O)Or~sC 2 -Clo-alkenyl, alkenyl as defined above, d) -(CH2)s-OI(C=O)OrsC 2 C6alkynyl, alkynyl as defined above, e) -(CH2)s0[I(C=O)Orls(C 3 -C 7 )-CYCloalky, 20 f) -(CH2)s-O[(C=O)Orjsary, aryl as defined above, g) -(CH2)s-O[(C=O)Orlsheteroaryl, heteroaryl as defined above, ago*h) -(CH2)s-O(CH 2 )flO(CH 2 )mheteroaryl, heteroaryl as defined above, i) -(CH2)s-O(CH2)flO(CH 2 )mary, aryl as defined above, j) -(CH 2 )s-OC(=O)NRlaR2, 25 k) -(CH 2 )s-OSO 2 R 3 1) -(Cl-C 6 )-alkyl, alkyl as defined above, or M) -(0 2 -C 6 )-alkenyl, alkenyf as defined above; R 2 9a and R29b together are =0 or =C(Cl-0 1 0 -alkyl) 2 alkyl being as defined above. 7. A compound selected from the group consisting of: 4,6,7,15,1 6-pentakis(acetyloxy)-21,22-epoxy-i 8-hydroxy-22-methoxycarbonyl[6a,7, 1 53, 1 6j, 2Oc.,21if,22 f]D:A-Friedo-A-homo2l 30-dinor-24-oxaoleana-li-en-3-one; 4,6,7,15,1 6-pentakis(acetyloxy)-2i,22-epoxy-i 8-hydroxy-22-methoxycarbonyl[6u.7a,15j3,1 6j, 2 0u.,21 f,22f]D:A-Friedo-A-homo2l 3Odinor-24-oxaoleana-i -ene; 4,6,7,15,1 6-pentakis(acety oxy)-20,21 ,22,(29)-diepoxy-i 8 -hydroxy-22-methoxycarbony[6L,7ct 1 6f,2Oct,21 f,22f]D:A-Friedo-A-homo-27,30-dinor-24-oxaoleana-li-en-3-one; [N:\LIBAAJO1 587.doc:TAB 113 4,6,7,15,1 6-pentakis (acetyloxy)-21I, 2 2 -epoxy- I 8-hydroxy-22-. methoxycarbonyl [6cx,7u, I Sf3,16j3,21lf, 2 2 f]D:A-Friedo-A-homo-27,30 dinor-24-oxaoleana. I -en-3 *29(Z)-methoxvcrbon-,Ai,7, 15,1 6paki loy-122eoy 10 2(E)methxycrbonl-46,7,5,1-pentakis(acetyloxy)..21, 2 2 -epoxy- 8 -hydroxy-22-methoxycarbonyl [6a,7oc, 15f,16f,21lf,221]D:AFriedo.. A-homo-2'7, 3 0 -dinor-24-oxaoleana.. I, 2 0(29)-dien-3 -one; 2 9 (E)-toxycarbonyl4,6715,1 -pentakis(acetyloxy)21 2 2 -epoxy- Il 8 -hydroxy-22-methoxycarbonyl [6ux,7oc, 15, 16f,21If,22f]D:A-Friedo.. I-oo2,0dno-4oala 1, 2 0 (29)-dien.3 -one; 2 9 (Z)-t-boxycarbonyl46715,1 6 -pentakis(acetyloxy2 1 2 2 -epoxy- hydrox-22-methoycrnyl6c,7uI 0,16,,20DAFre -homo-27 3 -dinor-24-oxaoleana. 2 0(29)-dien-3 -one; 2 9 (E)-et-boxycarbonyl4,6715,1 6 -pentakis(acetyloxy2 1 2 2 -epoxy- lhydroxy-22methoxycarbony [6Q,7cx, 5,163,2f, 2 23]D:A-Friedo homo-27 3 -dinor-24-oxaoleana 2 2 9 )-dien-3-one 2 9 (Z)-ethnoxycarbony,6,7 15,1 6 -petakis)-2,acetyoxy2 I 8 2 -epdoxy1- h2-drtoxy2..excarbonyIx [5, 6 ,x,15f,,2 lf3, 2 2f3]d:A-Freoo.-. hom0do-7,Onr-24-oleaa 1eal( 2 9 )-dien-3-one /V TO 114 2 cyano-46,7,15,1 6 -pentaki s(ace tylo xy)-2 I, 2 2 -epoxy-1 IX-hydroxy- 2 2 -methoxycarbonyl[6a,7Q, I 5J3,1 6f,21lt, 22 1]D:A-Friedo-A-homo-. 2 7 3 O-dinor-24-oxaoleana- I,2O(29)-dien-3 -one; 000 0 5 29(E)-phenyl-4,6,7, 15,1 6 -pentakis(acetyloxy)-2 I 2 2 -epoxy-i IS-hydroxy-. 000 2 2 -methoxycarbonyl[6x,7a, 1 53, 1 6j,21 0, 22 0]D:A-Friedo-A-homo- 2 7 3 O-dinor-24-oxaoleana- 1 2 O( 2 9)-dien-3-one; 29(Z)-phenyl-4,6,7, 15,1 6 -pentakis(acetyloxy-2 1 2 2-epoxy- I 8 1hydroxy- 2 2 -methoxycarbonyl[6a,7ux 1Sf, 1 6f,21 f,22f]D:A-Fredo-A-homo- 27,3 O-dinor-24-oxaoleana- I, 2 0(29)-dien-3 -one; 4 2 -bromobenizoyl)oxy-6,7, 15,1 6 -tetrakis(acetyloxy)..21I, 2 2 -epoxy- 18- 15hydroxy-22-rnethoxycarbony [6oc7cU. 5P,16$,20oc,21P3,22P13D :A 4,6,7,15,1 6 -pentaki s(acetyloxy)-21 2 2 -epoxy- I 8 -hydroxy-22-methoxy- carbonyl [6(x,7cx, 1 503,1603,20(x,210 f,220]D: A-Fri edo-Ahomo-27,3O dinor- 2 4 -oxaolea-3.-one; 4,6,7,15,1 6 -pentakis(acetyloxy)-21 2 2-epoxy- I 8-hydroxy-22- methoxycarbonyI[6oc,7cc 15P3, 1 6f,20oc,21 lf,22f3]D:A-FriedoAhomo. 27,3 O-dinor-24-oxaoleane; 4,6,7,15,1 6 -pentakis(acetyloxy)y2O(29),2 1,22-diepoxy- I 8-hydroxy-22. methoxycarbonyl[6(x7Qx 1 5P3,1 6f3,2Ox,2 1 f, 2 2 f]D:A-Friedo-A-homno 2 7 3 O-dinor-24-oxaoleana-en-3 -one; 4,6,7,15,1 6 -pentakis(acetyloxy)-21 2 2 -epoxy- I 8-hydroxy-22- methoxycarbonyl[6x7ax 153, 1 6f,2Ocx,21lf, 22 f3]D:A-Friedo-.A..homo-. 2 7 3 O-din or-24-oxaoleana-3,20-di one; 115 2 9 (Z)-methoxycarbonyl -4,6,7,15,1 6-pentakis(acetyloxy)-21I, 2 2 -epoxy. lX-hydroxy-22-methoxycarbonyl [6a,7ax, 1 51,1613,21 1,221]D:A-Fri-edo-. A -homo-27 3 O-dinor-24-oxaoleana.2O(29)-en-3-.one; 2 9 (E)-methoxycarbonyj -4,6,7,15,1 6 -pentakis(acetyloxy)-21I, 2 2 -epoxy- 8 -hydroxy-22-methoxycarbonyl [6a,7(x 1513,1613,2101,221]D:A-Friedo. A-homo-27 3 -dinor-24-oxaoeana.2(29)en-3-one; 2 9 (Z)-t-butoxycarbonyl-4,6,7, 15,1 6 -pentakis(acetyloxy-2 1 2 2 -epoxy- IX-hydroxy-22-methoxycarbonyl[6oc 7 a,1513,1613,21 1,221]D:A-Friedo- A C -homo-27,3-io-2-xoen-02)e- oe 2 9 (E)-t-butoxycarbonyl.4 15,1 6 -pentakis(acetyloxy-2 1 2 2 -epoxy- l 8 -hydroxy-22-meth ox ycarbonyl [6c,7c, 1513,1603,2101, 2 20]D: A-Frie do A-h omo 2 7 ,3-dinor24oxaoena2(29)en- 3 -oe 2 9 (Z)-ethoxycarbonyl46,7,15,1 6 -pentakis(acetyloxy)-21,2eo 8 hydroxy-22-methoxycarbonyl[6(x7c 1513,1613,2101, 2 21]D:A-Friedo-A homo- 2 7 ,3 Od inor24ox ao eana2(29)en-3-oe S 2 9(E)-ethoxycarbonyl-4,6,7,15,1 6 -pentakis(acetyloxy)-2 2 2 -epoxy- 18- hydroxy-22methoxycarbony[6x7xl 51,1613,2101,221]D:A-Friedo-A homo- 2 7 ,3O-d nor.24 o xaoeana2(29)en 3 -one 2 9(Z) -cyan o-4 ,6.7.15.1 -pe ntaki s(ace tyloxy)-21I, 2 2 -epoxy- I 8-hydroxy- 2 2 -methoxycarbonyIf 6(x.7Q, 1513,1613,21 1, 22 1]D:A-Friedo-.A..homo- 2 7 3 O-dinor-24-oxaoleana..20(29)en-3-one; 2 9 (E)-cyano-4,6,7, 15,1 6 -pentakis(acetyloxy-2 1 2 2 -epoxy- 1 8-hydroxy- 2 2 -methoxycarbonylf 6cx,7cx, 1513,1613,21 1, 22 1]D:A-Friedo-.A-homo. 273Odno-4oalen-02)e- oe 29(E)-phenyl4,6,715,1 6 -pentakis(acetyloxy-2 1 2 2 -epoxy- 1 8-hydroxy- 2 2 -methoxycarbonyl[6ox,7Q, 1513,1613,21 1, 2 2 1]D:A-Friedo..A.homo. 2 7 3 O-dior24oxaoeana2(29)en3one; 116 29(Z)-phenyl-4,6,7,15,16-pentakis(acetyloxy)-21,22-epoxy-18-hydroxy-22-methoxycarbonyl[6a,7a, 15p3,16p,21 P, 22 p]D:A-Friedo-A-homo-27,30-dinor-24-oxaoleana-20(29)-en-3-one; or 4-(2-bromo benzoyl)oxy-6,7,15,16-tetrakis(acetyloxy)-21,22-epoxy-18-hydroxy-22-methoxycarbonyl[6c,7, 16p,21 P, 22 p]D:A-Friedo-homo-27,30-dinor-24-oxaolean-3-one. 8. An immunosuppressant triterpene derivative, substantially as hereinbefore described with reference to any one of the examples. 9. A pharmaceutical formulation comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of the compound of formula I, as recited in any one of claims 1 to 8 or a pharmaceutically acceptable crystal form or hydrate thereof. lo10 10. The pharmaceutical formulation as recited in claim 9, comprising in addition, a second immunosuppressive agent comprising azathioprine, brequinar sodium, deoxyspergualin, mizaribine, mycophenolic acid morpholino ester, cyclosporin, FK-506 and rapamycin. S. 11. A method treating a condition in a mammal, the treatment of which is effected or facilitated by Kv1.3 inhibition, the method comprising the administration, in an amount that is effective 15 at inhibiting Kv1.3, of a compound as recited in any one of claims 1 to 8 or a pharmaceutical formulation as recited in claim 9 or claim 12. An embodiment which is a compound as recited in any one of claims 1 to 8 or a So pharmaceutical formulation as recited in claim 9 or claim 10 when used to treat a condition in a mammal, the treatment of which is effected or facilitated by Kv1.3 inhibition. 20 13. Use of a compound as recited in any one of claims 1 to 8 or a pharmaceutical formulation as recited in claim 9 or claim 10 in the manufacture of a medicament for treating a condition in a mammal, the treatment of which is effected or facilitated by Kv1.3 inhibition 14. The method as recited in claim 11, the embodiment as recited in claim 12 or the use as S recited in claim 13, wherein the condition is selected from the group consisting of: resistance by 25 transplantation of organs or tissue, graft-versus-host diseases brought about by medulla ossium transplantation; rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes uveitis, juvenile-onset or recent-onset diabetes mellitus, posterior uveitis, allergic encephalomyelitis, glomerulonephritis, infectious diseases caused by pathogenic microorganisms, inflammatory and hyperproliferative skin diseases, psoriasis, atopical dermatitis, contact dermatitis, eczematous dermatitises, seborrhoea dermatitis, Lichen planus, Pemphigus, bullous pemphigoid, Epidermolysis bullosa, urticaria, angioedemas, vasculitides, erythemas, cutaneous eosinophilias, Lupus erythematosus, acne, Alopecia areata, keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic keratitis, conical cornea, dystrophia epithelialis corneae, corneal leukoma, ocular pemphigus, Mooren's ulcer, Scleritis, Graves' opthalmopathy, Vogt-Koyanagi-Harada syndrome, sarcoidosis, etc; pollen allergies, reversible obstructive airway disease, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma and dust asthma, chronic or inveterate asthma, late asthma and airway hyperresponsiveness, bronchitis, gastric ulcers, vascular damage caused by ischaemic diseases and thrombosis, ischaemic bowel diseases, inflammatory bowel diseases, necrotising enterocolitis, 40 intestinal lesions associated with thermal burns and leukotriene B4-mediated diseases, Coeliac 1 N0 [N:\LIBAA]01587.doc:TAB O9 diseases, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease, ulcerative colitis, migraine, rhinitis, eczema, interstitial nephritis, Good-pasture's syndrome, haemolytic-uraemic syndrome, diabetic nephropathy, multiple myositis, Guillain-Barre syndrome, Meniere's disease, polyneuritis, multiple neuritis, mononeuritis, radiculopathy, hyperthyroidism, Basedow's disease, pure red cell aplasia, aplastic anaemia, hypoplastic anaemia, idiopathic thrombocytopaenic purpura, autoimmune haemolytic anaemia, agranulocytosis, pernicious anaemia, megaloblastic anaemia, anerythroplasia, osteoporosis, sarcoidosis, fibroid lung, idiopathic interstitial pneumonia, dermatomyositis, leukoderma vulgaris, ichthyosis vulgaris, photo allergic sensitivity, cutaneous T cell lymphoma, arteriosclerosis, atherosclerosis, aortitis syndrome, polyarthritis nodosa, myocardosis, 10 scleroderma, Wegener's granuloma, Sjogren's syndrome, adiposis, eosinophilic fascitis, lesions of S. gingiva, periodontium, alveolar bone, substantia ossea dentis, glomerulonephritis, male pattern alopecia or alopecia senilis by preventing epilation or providing hair germination and/or promoting hair generation and hair growth; muscular dystrophy; Pyoderma and Sezary's syndrome, Addison's disease, ischemia-reperfusion injury of organs which occurs upon preservation, transplantation or 15 ischaemic disease, for example, thrombosis and cardiac infraction, endotoxin-shock, pseudomembranous colitis, colitis caused by drug or radiation, ischaemic acute renal insufficiency, chronic renal insufficiency, toxinosis caused by lung-oxygen or drug, for example, paracort and bleomycins, lung cancer, pulmonary emphysema, cataracta, siderosis, retinitis, pigmentosa, senile macular degeneration, vitreal scarring, corneal alkali burn; dermatitis erythema multiforme, linear IgA 20 ballous dermatitis and cement dermatitis, gingivitis, periodontitis, sepsis, pancreatitis, diseases caused by environmental pollution, aging, carcinogenis, metastasis of carcinoma and hypobaropathy; disease caused by histamine or leukotriene-C 4 release; Behcet's disease, autoimmune hepatitis, primary biliary cirrhosis sclerosing cholangitis, partial liver resection, acute liver necrosis, necrosis caused by toxin, viral hepatitis, shock, or anoxia, B-virus hepatitis, non-A/non-B hepatitis, cirrhosis, 25 alcoholic cirrhosis, hepatic failure, fulminant hepatic failure, late-onset hepatic failure, "acute-on- chronic" liver failure, augmention of chemotherapeutic effect, preventing or treating activity of cytomegalovirus infection, HCMV infection, and antiinflammatory activity; and treatment of immunodepression or a disorder involving immunodepression, including AIDS, cancer, senile dementia, trauma, chronic bacterial infection, and certain central nervous system disorders. 15. The method, embodiment or use as recited in claim 14, wherein the condition is an autoimmune disease. 16. A method of preventing or treating the resistance to transplantation or transplantation rejection of organs or tissues into a patient, which method comprises the administration of a compound as recited in any one of claims 1 to 8 or of a composition as recited in claim 9 or claim 17. An embodiment which is a compound as recited in any one of claims 1 to 8 or a composition as recited in claim 9 or claim 10 when used to prevent or treat the resistance to transplantation or transplantation rejection of organs or tissues into a patient. 18. Use of a compound as recited in any one of claims 1 to 8 or a composition as recited in claim 9 or claim 10 in the manufacture of a medicament for preventing or treating the resistance to k'\transplantation or transplantation rejection of organs or tissues into a patient. [N:\LIBAA01587.doc:TAB 19. A method of suppressing the immune system in a subject, which method comprises the administration to the subject of an immune suppressing amount of a compound of formula I, as recited in any one of claims 1 to 8 or of a composition as recited in claim 9 or claim An embodiment which is a compound as recited in any one of claims 1 to 8 or a composition as recited in claim 9 or claim 10 when used to suppress the immune system in a subject. 21. Use of a compound as recited in any one of claims 1 to 8 or a composition as recited in claim 9 or claim 10 in the manufacture of a medicament for suppressing the immune system in a subject. 22. The method as recited in claim 19, the embodiment as recited in claim 20 or the use as 1o recited in claim 21, comprising the coadministration of a second immunosuppressive agent. 23. A method of preventing or treating resistance by transplantation of organs or tissue, graft-versus-host diseases brought about by a condition selected from the group consisting of medulla ossium transplantation; rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes uveitis, juvenile-onset or recent-onset 15 diabetes mellitus, posterior uveitis, allergic encephalomyelitis, glomerulonephritis, infectious diseases caused by pathogenic microorganisms, inflammatory and hyperproliferative skin diseases, psoriasis, atopical dermatitis, contact dermatitis, eczematous dermatitises, seborrhoea dermatitis, Lichen planus, Pemphigus, bullous pemphigoid, Epidermolysis bullosa, urticaria, angioedemas, vasculitides, erythemas, cutaneous eosinophilias, Lupus erythematosus, acne, Alopecia areata, 20 keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic keratitis, conical cornea, dystrophia epithelialis corneae, corneal leukoma, ocular pemphigus, Mooren's ulcer, Scleritis, Graves' opthalmopathy, Vogt-Koyanagi-Harada syndrome, sarcoidosis, etc.; pollen allergies, reversible obstructive airway disease, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma and dust asthma, chronic or inveterate asthma, late asthma and airway *i 25 hyperresponsiveness, bronchitis, gastric ulcers, vascular damage caused by ischaemic diseases and thrombosis, ischaemic bowel diseases, inflammatory bowel diseases, necrotising enterocolitis, intestinal lesions associated with thermal burns and leukotriene B4-mediated diseases, Coeliac diseases, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease, ulcerative colitis, migraine, rhinitis, eczema, interstitial nephritis, Good-pasture's syndrome, haemolytic-uraemic syndrome, diabetic nephropathy, multiple myositis, Guillain-Barre syndrome, Meniere's disease, polyneuritis, multiple neuritis, mononeuritis, radiculopathy, hyperthyroidism, Basedow's disease, pure red cell aplasia, aplastic anaemia, hypoplastic anaemia, idiopathic thrombocytopaenic purpura, autoimmune haemolytic anaemia, agranulocytosis, pernicious anaemia, megaloblastic anaemia, anerythroplasia, osteoporosis, sarcoidosis, fibroid lung, idiopathic interstitial pneumonia, dermatomyositis, leukoderma vulgaris, ichthyosis vulgaris, photo allergic sensitivity, cutaneous T cell lymphoma, arteriosclerosis, atherosclerosis, aortitis syndrome, polyarthritis nodosa, myocardosis, scleroderma, Wegener's granuloma, Sjogren's syndrome, adiposis, eosinophilic fascitis, lesions of gingiva, periodontium, alveolar bone, substantia ossea dentis, glomerulonephritis, male pattern alopecia or alopecia senilis by preventing epilation or providing hair germination and/or promoting hair A ,40 generation and hair growth; muscular dystrophy; Pyoderma and Sezary's syndrome, Addison's 0 [N:\LIBAAO1 587.doc:TAB \^NTO0 119 disease, ischemia-reperfusion injury of organs which occurs upon preservation, transplantation or ischaemic disease, for example, thrombosis and cardiac infraction, endotoxin-shock, pseudomembranous colitis, colitis caused by drug or radiation, ischaemic acute renal insufficiency, chronic renal insufficiency, toxinosis caused by lung-oxygen or drug, for example, paracort and bleomycins, lung cancer, pulmonary emphysema, cataracta, siderosis, retinitis, pigmentosa, senile macular degeneration, vitreal scarring, corneal alkali burn; dermatitis erythema multiforme, linear IgA ballous dermatitis and cement dermatitis, gingivitis, periodontitis, sepsis, pancreatitis, diseases caused by environmental pollution, aging, carcinogenis, metastasis of carcinoma and hypobaropathy; disease caused by histamine or leukotriene-C 4 release; Behcet's disease, autoimmune hepatitis, 10 primary biliary cirrhosis sclerosing cholangitis, partial liver resection, acute liver necrosis, necrosis S caused by toxin, viral hepatitis, shock, or anoxia, B-virus hepatitis, non-A/non-B hepatitis, cirrhosis, alcoholic cirrhosis, hepatic failure, fulminant hepatic failure, late-onset hepatic failure, "acute-on- chronic" liver failure, augmention of chemotherapeutic effect, preventing or treating activity of cytomegalovirus infection, HCMV infection, and antiinflammatory activity; and treatment of 15 immunodepression or a disorder involving immunodepression, including AIDS, cancer, senile dementia, trauma, chronic bacterial infection, and certain central nervous system disorders which comprises the administration of a compound as recited in any one of claims 1 to 8 or of a formulation as recited in claim 9 or claim 24. An embodiment which is a compound as recited in any one of claims 1 to 8 or a 20 composition as recited in claim 9 or claim 10 when used to prevent or treat resistance by S. transplantation of organs or tissue, graft-versus host diseases brought about by any one of the conditions recited in claim 23. Use of a compound as recited in any one of claims 1 to 8 or a composition as recited in claim 9 or claim 10 in the manufacture of a medicament for preventing or treating resistance by 25 transplantation of organs or tissue, graft-versus host diseases brought about by any one of the conditions recited in claim 23. 26. A method of treating a condition in a mammal, the treatment of which is effected or facilitated by Kv1.3 inhibition, the method comprising the administration of a pharmaceutical formulation comprising a pharmaceutical carrier and a compound of Formula I, as recited in any one of claims 1 to 8, in an amount that is effective at inhibiting Kv1.3. 27. An embodiment which is a pharmaceutical formulation comprising a pharmaceutical carrier and a compound of Formula I, as recited in any one of claims 1 to 8, in an amount that is effective at inhibiting Kv1.3 when used to treat a condition in a mammal, the treatment of which is effected or facilitated by Kv1.3 inhibition. 28. Use of a pharmaceutical formulation comprising a pharmaceutical carrier and a compound of Formula I, as recited in any one of claims 1 to 8, in an amount that is effective at inhibiting Kv.13 in the manufacture of a medicament for treating a condition in a mammal, the treatment of which is effected or facilitated by Kv1.3 inhibition. [N:\LIBAA]01587.doc:TAB 120 29. A method of treating a condition in a mammal, the treatment of which is effected or facilitated by Kv1.3 inhibition, the method comprising the coadministration of a therapeutically effective amount of a compound of formula I, as recited in any one of claims 1 to 8, with a second immunosuppressive agent. 30. An embodiment which is a compound of formula I, as recited in any one of claims 1 to 8 together with a second immunosuppressive agent when used to treat a condition in a mammal, the treatment of which is facilitated by Kv1.3 inhibition. 31. Use of a compound of formula I, as recited in any one of claims 1 to 8 together with a second immunosuppressive agent in the manufacture of a medicament for treating a condition in a 10 mammal, the treatment of which is facilitated by Kv1.3 inhibition. 32. A medicament when prepared by the method of any one of claims 13, 18, 21, 25 or 28. Dated 1 June, 1999 Merck Co., Inc. Patent Attorneys for the Applicant/Nominated Person SPRUSON FERGUSON 0 IN:\LIBAA]01587.doc:TAB