CA2863243A1

CA2863243A1 - Rafamycin analogs and methods for making same

Info

Publication number: CA2863243A1
Application number: CA2863243A
Authority: CA
Inventors: Bing Wang; Jonathon Zhong Zhao
Original assignee: HANGZHOU ZYLOX PHARMA Co Ltd
Current assignee: HANGZHOU ZYLOX PHARMA Co Ltd
Priority date: 2012-11-30
Filing date: 2012-11-30
Publication date: 2014-06-05
Also published as: EP2809675A4; WO2014082286A9; AU2012395673A1; ZA201405746B; AU2012395673A8; CN104854112A; JP2016500112A; WO2014082286A1; KR20150003156A; EP2809675A1; SG11201404432QA

Abstract

A semi-synthetic rapamycin analog with a triazole moiety or a pharmaceutically acceptable salt or prodrug thereof, is a broad-spectrum cytostatic agent and a m TOR inhibitor, and is useful in the treatment of various cancers, or tumors in organs such as kidney, liver, breast, head and neck, lung, prostate, and restenosis in coronary arteries, peripheral arteries, and arteries in the brain, immune and autoimmune diseases. Also disclosed are fungal growth-, restenosis-, post- transplant tissue rejection- and immune- and autoimmune disease- inhibiting compositions and a method of inhibiting cancer, fungal growth, restenosois, post-transplant tissue rejection, and immune and autoimmune disease in a mammal. One particular preferred application of such triazole-moiety containing rapamycin analog is in treating renal carcinoma, lung cancer, colon cancer, and breast cancers wherein potency of the drug, its half-life, tissue distribution properties, and its pharmacokinetic properties including bioavailability through oral and intravenous routes are essential to the clinical outcomes.

Description

RAFAMYCIN ANALOGS AND METHODS FOR MAKING SAME
BACKGROUND OF THE INVENTION
[OH The compound cyclosporine (cyclosporin A) has found wide use since its introduction in the fields of organ transplantation and immunomodulation, and has brought about a significant increase in the success rate for transplantation procedures. Recently, several classes of macrocyclic compounds having potent immunomodulatory activity have been discovered.
Okuhara et al.. in European Patent Application No. 184. 162, published Jun. I
I. 1986, discloses a number of macrocyclic compounds isolated from the genus Streptomyces, including the immunosuppressant FK-506. a 23-membered macrocyclic lactone. which was isolated from a strain of S. tsukuhaensis.
1021 Other related natural products. such as FR-900520 and FR-900523, which differ from FK-506 in their alkyl substituent at C-2 I. have been isolated from S.
hygroscopicus yakushimnaensis. Another analog, FR-900525. produced by S. tsukubaensis, differs from FK-506 in the replacement of a pipecolic acid moiety with a proline group.
Unsatisfactory side-effects associated with cyclosporine and FK-506, such as nephrotoxicity, have led to a continued search for immunosuppressant compounds having improved efficacy and safety.
including an immunosupressive agent which is effective topically. but ineffective systemically (U.S. Pat.
5A57.I I I ).
103] Rapamycin. as illustrated below, is a macrocyclic triene antibiotic produced by Streptomyces hygroscopicus. which was found to have antifungal activity, particularly against Candida alhicans. both in vitro and in vivo (US 3.929.992 and US 3.993.749).
1041 Rapamycin alone (US 4.885,171) or in combination with picibanil (US
4,401,653) has been shown to have antitumor activity. In 1977, rapamycin was also shown to be effective as an immunosuppressant in the experimental allergic encephalomyelitis model, a model for multiple sclerosis: in the adjuvant arthritis model, a model for rheumatoid arthritis:
and was shown to effectively inhibit the formation of IgE-like antibodies.
1051 The immunosuppressive effects of rapamycin have also been disclosed in FASEB in 1989, as has its ability to prolong survival time of organ gratis in histoincompatible rodents.
These and other biological effects of rapamycin are reviewed in Trunsplcmiution Reviews.. 1992.
6. 39-87. Mono-ester and di-ester derivatives of rapamycin (esterilication at positions 31 and 42) SUBSTITUTE SHEET (RULE 26) have been shown to be useful as antifungal agents (US 4.316.885) and as water soluble prodrugs of rapamycin (US 4,650,803).
1061 Mono-ester and di-ester derivatives of rapamycin (esterification at positions 31 and 42) have been shown to be useful as antifungal agents (US 4,316,885) and as water soluble prodrugs of rapamycin (US 4,650.803).
1071 Numerous chemical modifications of rapamycin have been attempted.
These include the preparation of mono- and di-ester derivatives of rapamycin (WO 92/05179). 27-oximes of rapamycin (FPO 467606): 42-oxo analog of rapamycin (US 5.023,262); bicyclic rapamycins (US
5.120.725): rapamvein dimers (US 5,120.727): silyl ethers of rapamycin (US
5.120.842); and arvIsullonates and sulfamates (US 5.177.203). Rapamycin was recently synthesized in its naturally occurring enantiomeric form (K. C. Nicolaou et al.. J. Am. Chem.
Soc., 1993, 115, 4419-4420:S. L. Schreiber. .1. Am. Chem. Soc.. 1993. 115.7906-7907: S. J.
DanishefskyõI. Am.
Chem. Soc.. 1993, 115. 9345-9346). One recent example of a rapamycin analog is a tetrazole containing rapamycin analog (US 6,015,815). The tetrazole heterocyclic ring is used to replace the hydroxyl group to effect the analog.
1081 Although some of these modified compounds exhibit immunosuppressive activity, anti-restenotic activities in suppressing the migration and growth of vascular smooth muscles.
especially when used in a stent coating, the need remains for rapamycin analogs which possess potentially enhanced efficacy against broad spectrum of cancers such as renal cell carcinoma, breast cancers, head and neck cancers, and potentially better lipophilicity.
longer half live in the blood or in local tissues. or resistance to oxidative forces and better stability in a formulation.
One way to achieve these goals is through introduction of a triazole moiety to the side chain of a rapamycin which may impart a better lipophilicity, better stability, better bioavailability, better tissue and cellular uptake. better efficacy compared to the known and existing modified rapamycin analogs or derivatives. The efficacy of the modified rapamycin may also have better potency against a variety of cancers, and potentially reduced toxicities.
SUMMARY OF THE INVENTION
1091 Accordingly. one object of the present invention is to provide novel semi-synthetic rapamycin analogs which possess a desired triazole moiety attached to either or both to 31C-, and or 42C-position of a rapamycin molecule.
SUBSTITUTE SHEET (RULE 26) 1101 In accordance with one aspect. the present invention is directed to compounds represented by the structural formula illustrated below.
[II] In accordance with one aspect. the present invention is directed to compounds represented by the structural formula illustrated below.
1121 In accordance with yet another aspect. a compound of the present invention may contain two such substitutes at both the 42C and 3 IC-positions of a rapamycin.
1131 The triazole moiety of the present invention may be introduced via a variety of reaction schemes, the typical ones are illustrated below:
, I 0 I , H OHt o- o A' Series A
wherein A' is one of the following structures:
NH
N \tr N=-=rc' ' , N . OH

N -N NN OH

A8 ,A9 A 0 OH
- N - .
N

N
N- N N,N0 N
All Al2 A13 A14 Series B:
SUBSTITUTE SHEET (RULE 26) =-=
N N¨

NJ N' N N = N N' IV > õ HN
N b N
(Th ,r4 R N- N N _N CON .N N N N
0 r 0 I., N N co,H N
- = N
H OH.
O ONR6R7Bg 89 1310 0 '0 N N N
N .N. N- N
0 OH N = = CO211 N ' 0 , N , = , = SOMH, N = CO/H
= r' ' 0 Another object of the present invention is to provide a synthetic processes for the preparation of such compounds from starting materials obtained by fermentation, as well as chemical intermediates useful in such synthetic processes.
[15] A
further object of the present invention is to provide pharmaceutical compositions containing, as an active ingredient. at least one of the above compounds.
1161 Yet another object of the present invention is to provide a method of treating a variety of disease states. including restenosis. post-transplant tissue rejection. immune and autoimmune dysfunction. fungal growth. and cancer, [17] In addition. the compounds of the present invention may be employed as an oral tablet.
oral solid or oral liquid, oral immediate or sustained release dosage, intravenous injection dosages. parenteral dosages. cream or solutions by formulation with pharmaceutically acceptable vehicles.

Also within the scope of this invention includes pharmaceutical compositions for immediate release or sustained release of its active ingredient. each comprising a compound of this invention and pharmaceutically acceptable excepient.

Still further related to this invention are medical devices, each comprising a compound of this invention. [xamples of the medical deices include drug-eluting coronary or peripheral.
esophageal. urinary. ovar, or neurovascular stem.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 Graph for Renal cell carcinoma tumor cell inhibition studies;
SUBSTITUTE SHEET (RULE 26) FIG. 2 Graph for Renal cell carcinoma tumor cell inhibition studies;
FIG. 3 Graph for Lung Cancer A549 cell inhibition studies;
FIG. 4 Graph for Lung Cancer A549 cell inhibition studies;
FIG. 5 Graph for Lung Cancer A549 cell inhibition studies;
FIG. 6 Graph for Melanoma SK-MEL-28 cell inhibition studies;
FIG. 7 Graph for Melanoma SK-MEL-28 cell inhibition studies;
FIG. 8 Graph for Melanoma SK-ME1.-28 cell inhibition studies:
FIG. 9 Graph for Epidermal cancer A431 tumor cell model;
FIG. 10 Graph for Epidermal cancer A43I tumor cell model:
FIG. 11 Graph for Epidermal cancer A431 tumor cell model:
FIG. 12 Graph for Glioblastoma U87 MG Tumor model studies;
FIG. 13 Graph for Glioblastoma U87 MG Tumor model studies;
FIG. 14 Graph for Glioblastoma U87 MG Tumor model studies;
FIG. 15 Graph for liuman colorectal tumor HCT 116 model studies;
FIG. 16 Graph for Human colorectal tumor HCT 116 model studies;
FIG. 17 Graph for Human colorectal tumor I ICI- 116 model studies;
FIG. 18 Graph for Breast cancer MDA-MB-231 tumor model:
FIG. 19 Graph for Breast cancer MDA-M13-231 tumor model:
FIG. 20 Graph for Breast cancer MDA-MB-23 I tumor model;
FIG. 21 Graph for Breast cancer mcr-7 tumor model;
FIG. 22 Graph for Breast cancer MCE-7 tumor model;
FIG. 23 Graph for Breast cancer MCF-7 tumor model;
FIG, 24 Graph for Prostate cancer PC-3 tumor studies:
FIG. 25 Graph for Prostate cancer PC-3 tumor studies;
FIG. 26 Graph for Prostate cancer PC-3 tumor studies:
SUBSTITUTE SHEET (RULE 26) FIG. 27 Efficacy of rapamycin analog of the present invention in treating HOT
116.
DETAIITD DESCRIPTION OF THE PREFERRED EMBODIMENTS
Definition of Terms 1201 The term "prodrug." as used herein. refers to compounds which are rapidly transformed in vivo to the parent compound of the above formula, for example, by hydrolysis in blood. A
thorough discussion is provided in T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery Systems." Vol. 14 of the A.C.S. Symposium Series, and in Edward B. Roche, ed., "Bioreversible Carriers in Drug Design." American Pharmaceutical Association and Pergamon Press, 1987.
both of which are hereby incorporated by reference.
1211 The term "pharmaceutically acceptable prodrugs." as used herein, refers to those prodrugs of the compounds of the present invention which are. within the scope of sound medical judgment. suitable for use in contact with the tissues of humans and lower mammals without undue toxicity, irritation, and allergic response. are commensurate with a reasonable benefit/risk ratio, and are effective for their intended use, as well as the zwitterionic forms, where possible. of the compounds of the present invention. Particularly preferred pharmaceutically acceptable prodrugs of the present invention are prodrug esters of the C-3I
hydroxyl group of compounds of the present invention.
1221 The term "prodrug esters." as used herein, refers to any of several ester-forming groups that are hydrolyzed under physiological conditions. Examples of prodrug ester groups include acetyl, ethanoyl, pivaloyl. pivaloyloNvmethyl. acetoxymethyl. phthalidyl.
methoxymethyl.
indanyl. and the like, as well as ester groups derived from the coupling of naturally or unnaturally-occurring amino acids to the C-31 hydroxyl group of compounds of the present invention.
1231 The term -isomer- as used herein, refers to a compound having the identical chemical formula but different structural or optical configurations.
1241 The term "epimer" as used herein, refers to a compound having the identical chemical formula but a different optical configuration at a particular position. In the case of a rapamycin, a 42-Epi rapamycin refers to the compound that has the opposite optical rotation compared to the rapamycin obtained by a fermentation process.
SUBSTITUTE SHEET (RULE 26) 1251 The term "15-isomer- as used herein, refers to the analog of rapamycin that contains a 7-member ring at the 15-position as opposed to a regular rapamycin obtained from a fermentation process which contains a six-member ring. This kind of conversion is also called "tautomerization-. The 15-isomer- as used herein. may also be referred to as a 15 tautomer of a rapamycin.
Preparation of Compounds 1261 The compounds and processes of the present invention will be better understood in connection w.t.1 the 2o. .ow .ng synthetic schemes w hich illustrate the methods by which the compounds of the present invention may he prepared.
1271 The compounds of the present invention may' be prepared by a variety of synthetic routes.
Most of the common conjugation reactions of rapamycin at 42- and/or 31-hydroxyl positions are found in the rapamycin patents mentioned above, the contents of which are incorporated herein by reference in their entireties.
EXAMPLES
Synthesis of Rapamyein Derivatives. The parent rapamycin structure is shown below.
4(4C) , \
'0 N

0 OH ,OH
0 Rapa Li- 0 Rap,' The synthetic scheme of series A of rapamycin analogs of the present invention is shown below:
-I
Y>, N A
Cin A I Series A
SUBSTITUTE SHEET (RULE 26) 1281 Shown below are additional rapamvcin analogs of this invention that were synthesized similarly:
wherein A. CO,H
NH
- N N N N N N N

.A5 .k3 OH

Q. '0 csi N=
"` NN N-N H

0 0H N N A7 A9 A10 "
- ,"
AN
-`y=OH N -NN
Series A NN NN
NN
k I I A 12 Al3 \14 Example 1: Synthesis of Compound Al 1291 To a stirred solution of Rapamycin (3 g. 3.2 mmol) and Cs2CO3 (3.2 g.
9.6 mmol) in dried WI' (90 ml.) \vas added Nat (1.5 g. 9.6 mmol) and 3-bromoprop-1-yne (1.2 g. 9.6 mmol).
The reaction mixture was stirred at rt for 30 hours. Upon the completion of reaction. 300 mt.
water was added in and extracted with ethyl acetate (200 m1_ x 3). The combined organic layer was washed by brine (300 ml.) and dried over anhydrous Na2SO4. After concentration, the residue was purified with silica gel chromatography (50% to 100% of ethyl acetate in petroleum ether as eluent) to give the compound Al (2.1 g. 68%) as a light green oil.
LCMS (m/z) ES- 950 (M-I )-.
Example 2: Synthesis of Compound A3 1301 "To a solution of 40-0-(prop-2-ynylox rapamycin A I (200 mi.:. 0.2 mmol) and 1-azido-Admantane (100 mg. 0.6 mmol) in anhydrous THE (9 ml.) was added DIPEA (100 af,. 0.6 mmol) and Cul (20mg. 0.1 mmol) under N,. The solution was stirred at rt overnight. Then, 20 water was added and extracted with ethyl acetate (20 ml. x 3). The combined organic layer \\ as washed by brine and dried over anh.\ drous Na2501. After concentration, the residue \\ as purified with silica gel chromatography (25% to 50% of ethyl acetate in petroleum ether as eluet) to give white solid which was further purified by prep-HPLC to give Compound A3 (26 mg.
10%) as a white solid. 'H NMR (300 MHz. CDC13) 6 7.71 (s, 1H), 6.74 (m, H), 6,39-6.02 (m.
5H), 5.62-5.36 (m. 510: LCMS (m/z)F.S- I I 28(M-1 ).
Example 3: Synthesis of Compound A4 SUBSTITUTE SHEET (RULE 26) 1311 To a solution of 40-0-(prop-2-ynyloxy) rapamycin Al (200 mg,. 0.2 mmol) and 4-azidobenzoic acid (100 mg. 0.6 mmol) in anhydrous THE (9 mL) was added D1PEA
(100 L.
0.6 mmol) and Cut (20 mg. 0.1 mmol) under N,. The solution was stirred at rt for 3 hours. Then.
20 ml. ater was added and extracted with ethyl acetate (20 nil. x 3). The combined organic layer was washed by brine and dried over anhydrous Na2804. After concentration. the residue was purified with silica gel chromatography (5% to 10% of methanol in dichloromethane as eluent) to give µYliite solid which was further purified by PREP-I IPI,C to give Compound A4 (29 mg. 12%) as a white solid. II NNIR (300 Ml ii. CDC13) 68.27 (m. III). 7.90 (m.
11-1). 7.73 (m.
I H). 7.56 (m. I H). 6.74 (m. 11-1). 6.55-6.00 (m, 51-1), 5.60-5.36 Om 5H):
LCMS (m/z) ES-11 14(M-1)1.
Example 4: Synthesis of Compound AS
Preparation of Intermediate 2 (110 NaN3, Tf2O, acetonitrile , rt, 2h N
a H2N 2 CuSO4, H20. TEA, rt, 6h N3 132] To a stirred suspension of NaN:, (2.0 g. 30.8 mmol) in acetonitrile (20 ml.) was added 1120 (7.3 g. 25.8 mmol) via syringe slowly at 0 C. The mixture was stirred for another 2 h at this temperature. The insoluble solids were removed through filtration. At 0 C. the filtrate was added dropwise into the mixture of Compound I (2.0 g, 13 mmol). CuSO4 (160 mg.
1 mmol).
H,0 (6 ml..) and Ft3N (3.6 mL. 25.8 mmol). The reaction mixture was stirred for 6 h at room temperature. The mixture was diluted with Et0Ac and washed with brine. The organic layer was dried over Na2SO4 and evaporated to yield brown solid which was purified with silica 2e1 chromatography (30% to 50% of 110Ac in petroleum ether as eluent) to give Intermediate 2 (1.1 g. 48%) as a w lute solid. 11-1 NMR (300 Mlii,. CDC1,1) 6 9.93 (s. III). 7.63 (m. 21I). 7.03 (m. 211).
2.02 (s. 311): I.CMS (m/z) F.S4- 177 (NI+ 1 1.331 To a solution of 40-0-(prop-2-ynyloxy) rapamycin A I (200 mg. 0.2 mmol) and N-(4-azido- phenyl) acetainicle. Intermediate 2 ( I 00 nm. 0.6 mmol) in anhydrous THE (9 ml.,) was added Dl PEA (100 1.., 0.6 mmol) and Cul (20 tug. 0.1 mmol) under N,. The solution was stirred at rt for 4 hours. Then. 20 mL. water was added and the mixture was extracted with Et0Ac (20 ml, x 3). The combined organic layer was washed by brine and dried over anhydrous Na2SO4.
SUBSTITUTE SHEET (RULE 26) Alter concentration. the residue was purified with silica gel chromatography (30% to 100% of Et0Ac in petroleum as eluent) to give white solid which was further purified by prep-HPLC to give Compound AS (56 mg, 25%) as a white solid. 1H NMR (300 MHz, CDCI3) 6 8.13 (m, 1H), 7.73 (m. 41-1), 6.74 (m, 6.49-6.00 (m. 511), 5.65-5.37 (m, 5H); LCMS (m/z) ES- 1127 (M-1)-.
Example 5: Synthesis of Compound A6 R
TMSN3. Na2CO3. Fri 0 H
OuS0,. sodium ascorbate R - TF3AF THE- it, 7h ^
t-BuOH. H20 rt. 3h "o-N TMS
' Al A2NNA6 1341 To a solution of 40-0-(prop-2-ynyloxy) rapamycin A I (200 mu. 0.2 mmol) and TMS-(IOU mg. 0.9 mmol) in t-13u011 (6 mi.) and 1-120 (6 ml.) was added Na2CO3 (100 mg, 1 mmol).
CuSO4 (20 mg, 0.13 mmol) and sodium ascorbate (40 mg, 0.2 mmol) under N,. The solution was stirred at rt for 3 hours. Then. 20 mL water was added and extracted with Et0Ac (20 mL x 3). The combined organic layer was washed by brine and dried over anhydrous Na2SO4. After concentration, the residue was purified with silica t.,,e1 chromatography (25%
of Et0Ac in petroleum ether as eluent) to give compound A2 (189 mg, 82%) as a white solid which was dissolved in IBM' in THF (10 ml.) at 0 C and stirred at rt for 7 hours. Then the reaction mixture was partitioned between Ft0Ac and water. The aqueous phase was extracted with Lt0Ac (25 ml. x 3). The combined organic laver was \\ ashed with brine and dried over anhydrous Na2S0.4. After concentration. the residue was purl tied with silica gel chromatography (5% to 10% of methanol in dichloromethane as eluent) to give white solid which was further purified by prep-FIPLC to give compound A6 (38 mg, 24%) as a white solid. 1H
NMR (300 MHz.
CDCI3) 6 7.75-7.55 (m. 114 6.76 (m, 1 I-1). 6.49-6.08 (m. 511). 5.53-5.35 (m, 3H); LCMS (m/z) ES- 1012 (M-1-f-18)'.
Example 6: Synthesis of Compound A7 Preparation of ntermed kites 5 and 6:
Br N HO-NaN3, H20 TsCI, TEA Cs2003, morpholine 1,-refluxing, DCM. it, 4h DMF, it, overnight overnight 6 SUBSTITUTE SHEET (RULE 26) 135i To a solution of 3 ( 5 u. 41 mmol) in 100 ml.. of water was added NaN1 (5g. 83 mmol) and was refluxed overnight. Then 100 ml. [)CM was added in after reaction mixture was cooled to rt. The organic phase separated was dried over Na2S0.1. filtered. To the solution was added Et3N (5.05 u. 50 mmol) and IsCI (9.55 g. 50 mmol) at 0 deg. The reaction mixture was stirred at rt for 4 hours. 100 ml.water \\ as added. The organic phase was separated and dried over Na2S0.2.
Filtration and concentration in vacuo gave the crude product. Purification by column chromatography (10% of Et0Ac in petroleum ether as eluent) gave intermediate 5 (5.7 g, 58%) as a colorless oil. IFl NMR (300 MHz, CDC13) 6 7.81 (d. 2H), 7.39 (d, 211).
4.14 (m,2H), 3.48 (m.
2I-1).2.43(s. 3H): I.CMS (m/z) FS+ 242(M+1)-'.
1361 To a solution of intermediate 5 (I g. 4,1 mmol) and Cs2CO3 (2.8 g. 8.2 mmol) in 30 ml.
of anhydrous DM': was added morpholine (0.71 g. 8,2 mmol) at 0 C. Then it was stirred at rt overnight. The reaction mixture \\ as partitioned between 50 ml of Ft0Ae and 60 mL of water.
The organic phase was dried over Na?S0.4. Illtration and concentration in VaCLIO gave the crude product. Purification by column chromatography (50% of It0Ac in petroleum ether as eluent) gave intermediate 6 (0.4 u. 72%) as a colorless oil. [.CMS (m/i.) FS 4 157 (M4-1)'.
1371 To a solution of 40-0-(prop-2-ynylox) rapamycin AI (200 mg. 0.2 mmol) and 4-(2-azido- ethyl)morpholine. Intermediate 6 (100 mg, 0.6 mmol) in anhydrous THE (9 mL) was added DIPEA (100 L. 0.6 mmol) and Cu! (20 mg, 0.1 mmol) under N-). The solution was stirred at rt for 3 hours. Then. 20 ml. water was added and extracted with Et0Ac (20 mL x 3). The combined organic laver was washed by brine and dried over anhydrous Na2SO4.
After concentration, the residue \\ as purified with silica gel chromatography (30%
of Nt0Ac in petroleum ether as eluent) to give white solid which was further purified by prep-HPLC to give compound A7 (45 mg. 20%) as a white solid. IF! NMR (300 Ml ii. CDCL,) 6 7.89 (m. 1H). 6.72 (m. 1 II). 6.44-6,05 (m. 511). 5.60-5.37 ( m. 511): [('MS (m/i) [5- 1107 (NI-1 f.
Example 7: Synthesis of Compound A9:
1381 To a solution of 40-0-(prop-2-ynyloxy) rapamycin A 1 (200 mu. 0.2 mmol) and 2-azidoethanol (100 mg. 1.2 mmol) in anhydrous TI IF (9 mL) was added DIPEA (100 L. 0.6 mmol) and Cul (20 mg. 0.1 mmol) under The solution was stirred at rt overnight. Then, 20 ml.. water was added and extracted with Et0Ac (20 ml, x 3). The combined organic layer was washed by brine and dried over anhydrous Na-,.S0.1. After concentration. the residue was purified SUBSTITUTE SHEET (RULE 26) with silica gel chromatography (5% to 10% of methanol in dichloromethane as eluent) to give white solid which was further purified by prep-HPLC to give compound A9 (26 mg, 11%) as a white solid. IH NMR (300 MI lz. CDCI3) 6 8.03-7.78 (m. 1H), 6.70 (m, I H), 6.46-6.00 (m. 5H).

5.61-5.39 (m, 511): I.CMS (m/i.) FS- 1038 (M-1)-.
Example 8: Synthesis of Compound A10:
Preparation of Intermediate 8 F-0 HBr, rt, 1 h NaN3, DMSO, Br 80deg, 2 days N3 HO
OH OH

solution of compound 7 (1.3 g. 12.7 mmol) in 40% HBr (10 mL) was stirred at rt for I
hour. Then. 20 mL water was added and extracted with Ft0Ac (20 mL x 3). The combined organic laver was washed by brine, dried over anhydrous Na2SO4. After concentration, the residue was purified with silica gel chromatography (50% of Et0Ac in petroleum ether as eluent) to give intermediate 8 (0.7 g. 31%) as a white solid.
I.CMS (m/z) LS+ 183 (M
Preparation of Intermediate 9 solution of' intermediate 8 (0.7 g. 3.9 mmol) and NEIN.; (1.13 g. 15 mmol) in DMS0 (16 ml..) was stirred at 80 deg for 2 days. Then. 20 mt. water was added and extracted with Et0Ac (20 nil. x 3). The combined organic layer was washed by brine, dried over anhydrous Na250.1.
After concentration, the residue \vas purified with silica gel chromatography (50% to 100% of Ft0Ac in petroleum ether as eluent) to give intermediate 9 (0.25 g. 45%) as a white solid.
LCMS (m/z) ES+ 146 (M-1.1)'.
Preparation of Compound A 10 1411 to a solution of' 40-O-( prop-2-ynyloxy) rapamycin A I (200 mg. 0.2 mmol) and 2-(azidomethyl) 2-methylpropane-1.3-diol Intermediate 9 (100 mu. 0.7 mmol) in t-Bu011 (6 m1.) and H20 (6 ml.) was added Na2CO3 (100 mg. 1 mmol), CuSO4 (20 mg, 0.13 mmol) and sodium ascorhate (40 mu. 0.2 mmol) under The solution was stirred at rt for 6 hours. Then. 20 nil.
water was added and extracted \\ ith Et0Ac (20 ml. x 3). 'Hie combined organic layer was washed by brine and dried over anhydrous Na250.1. After concentration, the residue was purified with silica gel chromatography (5% to 10% of methanol in dichloromethane as eluent) to give SUBSTITUTE SHEET (RULE 26) while solid w hich was further purified 11\ prep-IIPI.0 to give compound A10 (15 mg. 7%) as a white solid. I H NMR (300 Nil It. cDci,) 6 7.76 (m. III). 6.69 (m. 111), 6.55-6.00 (m, 511). 5.63-5.33 (m. 511). LCMS (m/z) ES- 1096 (M-1)-.
Example 9: Synthesis of Compound Al2 Preparation of Intermediate II
Li0H, THE, MeON
0 ' 0 H20, rt, 3h 1421 To a mixture of compound 10 (I 2. 7.8 mmol) in of Me01-1/TIIF (10 mL/I
0 iii L) was added a solution oftAA 1 (0.9 LI. 39 mmol) in 10 ml of w.ater. The resulting solution is stirred at room temperature for 3 hours. I he mixture was acidified bv 2N 11(1 to PH -4.
and extracted with Et0Ac (25 nil. ,,2). The combine organic laver was concentrated under vacuum to give intermediate 11 (0.7 g, 910/0) as a colorless oil. 11-1 NMR (300 MHz, CDC13) 6 2.34 (s,2H):
LCMS (m/z) ES+ 102 (M+1)'.
Preparation of Compound Al2 1431 To a solution of 40-0-(prop-2-ynyloxy) rapamycin A 1 (200 mu. 0.2 mmol) and 2-azidoacetic acid Intermediate 11 ( IOU mg. I mmol) in t-1.3u0H (6 ml,) and 110 (6 ml,) was added Na:CO3 (100 mg. 1 mmol). CuSO4 (20 mg. 0.13 mmol) and sodium ascorbate (40 mg, 0.2 mmol) under N. The solution was stirred at rt for 2 hours. 'Ben. 20 ml water was added and extracted \\ id) FIOAc (20 3). 'Hie combined organic layer was washed by brine and dried over anhydrous Na:SO4. After concentration. the residue was purified with silica gel chromatography (5% to 200/0 of methanol in dichloromethane as eluent) to give white solid which was further purified by prep-HP1.0 to give compound Al2 (15 mg. 7%) as a white solid.
H NMR (300 MHz. CDCI3) 6 7.89 (m. I H). 6.72 (m, 1H). 6.49-6.08 (m. 5H), 5.60-5.35 (m, 511):
LCMS (m/z) ES- 1052 (M-1).
Example 10: Synthesis of Compound A13 Preparation of intermediate 13 SUBSTITUTE SHEET (RULE 26) \N_ 1 NaN3, Tf20, acetonitrile , it, 2h 2 CuSO4, H20, TEA, it, 6h 1441 to a stirred suspension of NaN,, (2.0 g. 30.8 mmol) in acetonitrile (20 mL) was added ILO (7.3 g. 25.8 mmol) s\
rinee SIO\\ 1\ at 0 deg.. The mixture was stirred for another 2 h at this temperature. The insoluble solids wel'e. removed through filtration. At 0 deg. the filtrate was added dropwise into the mixture of compound 12 (2.0 g, I() mmol), CuSO4 (160 mg, 1 mmol).
11,0 (6 nil.) and Ft3N (3.6 ml.. 25.8 mmol). The reaction mixture was stirred for 6 11 at room temperature. The mixture was diluted with Et0Ac and washed with brine. The organic layer was dried over Na2SO4 and evaporated to yield brown solid which was purified with silica gel chromatography (30% to 50% of Ft0Ac in petroleum ether as eluent) to give brown solid which was further purified by prep-I IPEC to give intermediate 13 (0.4 LI. 17%) as a brown solid. 111 NMR (300 MI It. CDCL) 6 6.94 (m, 411). 3.19 (m. 41I). 2.60 (m. 411). 2.36 (s.
ECMS
(m/z)ES4- 218 (M1 I )'.
Preparation of Compound Al3 1451 In a solution or 40-0-(prop-2-ynylox rapamycin Al (200 mg,. 0.2 rnmol) and 1-(4-azido- pheny1)-4-methylpiperaiine Intermediate 13 (100 mg. 0.5 mmol) in t-Bu01-1 (6 inE) and 1120 (6 111E) was added Na2CO3 (100 mg. 1 mmol), CuSO4 (20 mg, 0.13 mmol) and sodium ascorbate (40 mg. 0.2 mmol) under N,. The solution was stirred at rt for 3 hours. Then, 20 niL
water was added and extracted with Ft0Ac (20 ml. x 3). The combined organic layer was washed by brine and dried over anhydrous Na2SO4. Alter concentration. the residue was purified with silica gel chromatography (5% to 20% of methanol in diehloromethane as eluent) to give white solid which was further purified by prep-111TC to give compound Al3 (33 mg. 14%) as a white solid. '11 NMR (300 MI It. CDCI;) 6 8.11 (m. 111). 7.71 (m. 211). 7.06 (m. 214 6.70 (m.
111). 6.45-6.00 (m. 511). 5.66-5.36 (m. 511): I.CMS (m/i.) ES- 1168 (M-1)-.
Example 11: Synthesis of Compound Al4 1461 To a solution of 40-0-(prop-2-ynyloxy) rapamvcin AI (200 Inv,. 0.2 mmol) and 1-(azido-methyl)-4-fluorobenzene (100 mg. 0.6 mmol) in anhydrous TI IF (9 mL) was added D1PEA (100 E. 0.6 mmol) and Cut (20 mg. 0.1 mmol) under N,. The solution was stirred at rt for 3 hours.
SUBSTITUTE SHEET (RULE 26) Then. 20 ml, water \\. as added and extracted with Pt0Ae (20 ml. x 3). The combined organic layer as washed by brine and dried over anhydrous Na2SO4. Alter concentration, the residue was purified with silica gel chromatography (30% to 100% of Et0Ac in petroleum ether as eluent) to give white solid which was further purified by prep-11PLC to give compound A 14 (32 mg. 14%) as a white solid. '11 NMR (300 MHz. CDCI3) 6 7.58 (m. 1 LI). 7.26 (m.
211). 7.06 (m.
211), 6.75 (m. 1 II). 6.50-6.00 (m. 511). 5.60-5.36 (m. 511): I.CMS(m/z)FS--1102(M-1).
Example 12: Synthesis of Compound A15 TBDPSCI, TEA, DMAP DIEA. 1120, DCM
TBDPS
DCM, rt, overnight rt, overnight RO H
16, DIEA, toluene, R HF/pyridine. THF C) \>", 'OH 80 deg, 2h rt, 4h Preparation of Intermediate 15 1471 to a solution of compound 14 g.
-18 mmol) and kt (5 g. 50 mmol) in DCM (100 m1.) \vas added DMA P (0.6 g. 5 mmol) and drop w ised TI3DPS-C1 (4.4 g. 16 mmol) at 0 deg and stirred at rt overnight. Then. I 00 1111, water was added and extracted \vith DCM (80 m L x 3). The combined organic laver \\ as washed by brine and dried over anhydrous Na,SO4.
After concentration, the residue \\ as purified \\ ith silica gel chromatography (30% of Et0Ac in petroleum ether as eluent) to give intermediate 15 (1.7 g. 12%) as a colorless oil. 111 NMR (300 MI lz. CDCI3) 6 7,63 (lit -1r1). 7.36 (m. 6H), 3.74 (m.21-1), 3.66 (m, 211), 1.04 (s.91-1); LCMS (m/z) ES+ 301 (M+1)'.
Preparation of Intermediate 16 1481 To a solution of intermediate 15 (1.7 LI. 5.7 mmol) and Dl PEA (1.5 g.
11.4 mmol) in DCM (40 ml.) was added 1 10 (1.7 g. 6 mmol) at 0 deg and stirred at rt overnight. Then. 50 m1.
water was added and extracted \\ ith DCM (40 ml. x 3). The combined organic layer was washed by brine and dried over anhydrous Na:SOA. After concentration, the residue was purified with silica gel chromatograph ( of Ft0.Ac in petroleum ether as cluent) to give intermediate 16 SUBSTITUTE SHEET (RULE 26) (1.5 g. 63%) as a colorless oil. 11 NMR (300 MHz. CDCF) 6 7.63 (m. 411). 7.36 (m. 611). 4.58 (m,211). 3.95 (m. 211). 1.04 (s.91I): 1,CMS (m,/z) ES-I- 433 (M+1)..
Preparation of Intermediate 17 1491 .lo a solution of Rapamycin (400 mg. 0.43 mmol) and DIPFA (278 mg.
2.15 mmol) in toluene (30 ml was added intermediate 16 (0.93 g. 2.15 mmol) at rt and stirred at 80 deg for 2 hours. Then. 50 ml, water was added and extracted with 1/t0Ac (30 m1_, x 3).
The combined organic layer was washed by 0.5 N 1-ICI, saturated NaHCO3 and brine, dried over anhydrous Na2SO4. After concentration, the residue was purified with silica gel chromatography (25% to 40% of Ft0Ac in petroleum ether as cluent) to give intermediate 17 (280 mg.
53%) give as a \\Ilite solid. 1.CMS (iniz) Es- I 94 (M-11.
Preparation of Compound Al5 1501 To a solution of intermediate 17 (280 mg. 0.23 mmol) in THE (10 ml_.) was added 2 ml.
HE in pyridine at 0 dog and stirred at rt for 3 hours. Then. 20 ml water was added and extracted with It0Ac (20 nil. x 3). Ihe combined organic la\ or was washed b) 0.5N 11C1.
saturated NalICOt and brine, dried over anhydrous NaSO4. Alter concentration, the residue was purified with silica gel chromatography (30% to 100 % of Ft0Ac in petroleum ether as eluent) to give white solid which was further purified by prep-I !PLC to give compound Al5 (34 mg. 15%) as a white solid. 111 NMR (300 MI Ii. CDCF) 6 6.40-6.00 (m. 511). 5.53-5.25 (m.
4H), 4.83 (s. 11-1).
4,13 (m. I.CMS (m/z) FS- 957 (M-1)-.
Synthesis or series 13 of rapamvcin derivatives oldie present invention 1511 13 series of rapam. cin derivati\ es \\ ere prepared according to the rollo\\ ing reaction scheme:
R 0 Tr-i0NaN R - 0 - =
'OH DCM
'OTf NN.I3 'Click Reaction' Rapamycin Compound I3 Compound III
Series [3 1521 In the formula of the schemes shown above. R and 13' have the following structures in some examples of the 13 series of rapamycin derivatives.:
SUBSTITUTE SHEET (RULE 26) HQ
, f -OH
N N. N - =
N N, NN N¨

R 13' = N 'OH N- 4 N N õ:" HN
-kO (!),,,, - --N

.., -,1 H ' N-- N N NN N ---N --N
9 - 0 NA N N'N, -0 oh , / N

I

N- N -NJ NN N N-N " ' CO2H N- .
_ SO2N112 N -_ ---, - SO2Nfi2 N ,--- CO7H
N 0 , : , Example 13: Synthesis of Compound 131 R0T0 R NaN3 ..Ø0 R 0 f2 -, ______________________ .. _______________________ t 'OH DCM "OTf 4...CICN:
compound B compound B1 1531 lo a solution or !warm chi (5 ti,. 5.5 mmol) and 2.6-di-tert-buty1-4-meth lpyridine (3.4 g.
3.4 mmol) in dried DCM (150 ml.). trilluoromethanesullonic anhydride (1.55 g.
5.5 mmol) was added at 0 oC. Alter the mixture was stirred for 2 h at room temperature. NaN3 (3.6 g, 55 mmol) was added and [WS() (60 ml ) was added at -10 oC. the mixture \\ as stirred Col- at 40 oC lor 5 h.
The mixture \\ as quenched 1-).\ addition of water. extracted \\ ith DCM (200 ml. x 2). and the combined extracts were washed \\ ith water. dried over Na2S01. and evaporated to dryness in vacuo. The crude product was purified by column chromatography (25% of Et0Ac in petroleum ether as eluent) to give compound B (1.5 g. 300/) as a white solid. LCMS (m/z) ES- 937 (M-HY.
Example 14: Synthesis of Compound B2 0¨

compound B1 ,0.---7-'0H - - R /----c_. /-------(OH
CuSO4 =-\
/ N-N
Na2CO3 'N-SM1 Me0H/H20 Compound B2 1541 to a solution of compound li 1 (ISO nut. 0.16 mmol) and SMI (27 mg. 0.48 mmol) in Me011/11,0 (4 ml 12 ml.) was added vitamine C sodium salt (63 mg. 0.32 mmol).
followed \\ ith the addition of. CuSO4 (51 mg. 0.32 mnfol) and Na:,.001 (51 mt.t. 0.48 mmol).
Aker stirred overniat, the mixture \\ as littered. the filtrate \\ as concentrated and purified by column SUBSTITUTE SHEET (RULE 26) chromatography (0 to 2% of methanol in dichloromethane as cluent) to give Compound 132 (33.2 mg. 21%) as yellow solid. II
N (300 MIlt. CD(13) 6 7.83 (m. 111). 6.37-6.01 (m. 41-1). 5.40-5.31 (m. 411): ELMS (W') ES- 093 (M-I
Example 15: Synthesis of Compound 133 compound B1 Na2CO3 R"-<
CuSO4 vitamine C sodium salts compound B3 1551 To a solution of compound 131 (150 rrq..!õ 0.16 mmol) and SM1 (40 mg. 0.48 mmol) in Me011/1+0 (4 ml /2 ml \\ as added vitaminc C sodium salt (63 11144. 0.32 mmol) followed \\ ith the addition of CuS0.1 (51 mg. 0.32 mmol) and Na2C0t (51 mg. 0.48 mmol). After stirred overnight. the mixture \\ as filtered. the filtrate was concentrated and was purified by column chromatography (0 to 2% of methanol in dichloromethane as cluent) to give Compound 133 (20.7 mg. 13%) as \\ hitc solid. 'II NM R (300 M117. CDCE) 6 8.55 (s. 11). 7.80 (m.
111). 6.39-6.02 (m.
411). 5.46-4.83 (m. 411): ELMS (m/z) ES- 1020 Example 16: Synthesis of Compound 114 o- OH
DCMcompound B1 E3r H2N
0"C 1h Step 1 Step 2 SM1 1 2 compound 84 1561 To a solution of I (2 g. 32.8 mmol) in DCM (100 ml.) was added SM1 (2 g. 16.4 mmol) at 0 oC dropw ise over I hour. rhe mixture \Vas concentrated and the residue was purified by column chromatograph\ (0 to 200 of methanol in dichloromethane as eluent to giVe intermediate 2 (0.0 (2. 54%) as yellow oil. ELMS (m/z) [54- 100 NH
1571 To a solution of compound 131 (150 mg. 0.16 mmol) and intermediate 2 (48 mg,.
0.48 mmol) in Me011/1120 (4 ml ./2 ml,) was added vitamine C sodium salt (63 mg.
0.32 mmol) followed with the addition of CuSO4 (51 mg. 0.32 mmol) and Na2CO3 (51 mg, 0.48 mmol).
After stirred overnight, the mixture was filtered, the filtrate \\ as concentrated and purified by column chromatOgraphy (0 to 2% of methanol in diehloromethane as eluent) to give Compound 134 (17.1 mg. I 1"/0) as white solid. 11 1 NMR (300 lV111/. ('DCE) 6 8.40 (s.
111). 6.39-6.02 (m. 411).
5.37-4.94 (m. 411): ELMS (m/i) ES- 1036 (M-11)-.
SUBSTITUTE SHEET (RULE 26) Example 17: Synthesis of Compound 135 N OH compound 61 0--. H K2CO3 1- 1, õOHNOH
1111L0F1 Step 2 10--\
Step 1 SM1 2 compound B5 1581 To a solution of I (2 u. 19.0 0111101) in TI IF (40 ml .1 was added Kr,CO3 (5.2 g, 38 mmol) and SM I (2.2 g. 19 mmol ) at 0 C. After stirred or 4 h at r.t. the mixture was filtered, the filtrate was concentrated and purified by column chromatography (0 to 3% of methanol in dichloromethane as eluent) to give intermediate 2 (1.2 g, 44%) as yellow oil.
11-1 NMR (300 Mfiz.
CDC1.1) 63.65 (m. 411). 3.49 (m. 21-1), 2.75 (m. 41-1), 2.22 (m.
1591 'lo a solution of compound B 1 (150 01g. 0.16 mmol) and intermediate 2 (69 mg, 0.48 mmol) in Me01-1/1120 (4 ml .12 ml.) was added vitamine C sodium salt (63 mg, 0.32 mmol) followed with the addition of CuSai (51 mu. 0.32 mmol) and Na2CO3 (51 mg, 0.48 mmol).
After stirred overnight. the mixture was filtered, the filtrate was concentrated and purified hy column chromatography 10 to 2% of methanol in dichloromethane as eluent) to give Compound 135 (26.7 mg. 11%) as white solid. 11 1 NMR (300 MI lz. C'DCI3) 6 7.92 (s, 111). 6.39-5.99 (m.
411). 5.45-4.84 (m. 411): I,Cl\/1S (m/z) 1080 (M-I If .
Example 18: Synthesis of Compound B6 cs2co, compound B1 THF Step 2 RNNO
Step 1 SM1 1 2 compound 66 1601 To a solution of 1(0.5 g. 5.8 mmol) in TI IF (40 ml.) was added Ki,CO3 (1.6g. 11.6 mmol) and SM I (0.69 i, 5.8 mmol) at 0 oC. Alter stirred for 4 h at It. the mixture was filtered, the filtrate \\ as concentrated and purified by column chromatography (3% of methanol in dichloromcthane as eluent) to give 2 (0.3 u. 42%) as yellow oil. [CMS (m/z) [S+ 125 (M+1-I) 1611 lo a solution of compound 111 (ISO mg. 0.16 11111101) and 2 (60 mu.
0.48 mmol) iii Me01 I/1120 (4 101/2 ml,) \\ as added vitamine C sodill111 SaIl (63 fig. 0.32 mmol) followed \\ ith the addition of CuS01 (51 mg. 0.32 mmol) and Na:,CO: (51 mg. 0,48 mmol). After stirred oyerniuht, the mixture \\ as filtered. the filtrate \vas concentrated and purified by column chromatography (0 to 2% of methanol in dichloromethane as eluent) to give Compound 136 (25.8 SUBSTITUTE SHEET (RULE 26) mg. 15%) as white solid. if1 NMl (300 MI lz. CDCI3) 6 7.81 (s. III), 6.40-6.02 (m, 41-1). 5.45-4.81 (m. 411): 1.CMS (mi.) [5- 1062 (M-11)-.
Example 19: Synthesis of Compound 137 compound B1 0¨

Cs2CO3 THF Step 2 N N
Stop 1 SM1 1 2 compound B7 1621 To a solution of 1 (0.5 u. 5 mmol) in T111: (40 ml.) was added K.2CO3 (1.4 g. 10 mmol) and SM I (0.6 g. 5 mmol) at 0 C. After stirred for 4 h at r.t. the mixture was filtered, the filtrate \vas concentrated and purified by column chromatography (3% of methanol in dichloromethane as eluent) to give intermediate 2 (0.5 g. 72%) as \ am\ oil. I.(MS (m/z) LS+
139 (M4-11)'.
1631 to a solution of compound 131 (150 mu. 0.16 mmol) and 2 (60 mu. 0.48 mmol) in Me011/1120 (4 m112 ml,) was added vitamine C sodium salt (63 mu. 0.32 mmol) followed with the addition of CuS0.4 (51 mu. 0.32 mmol) and Na2CO3 (51 111g, 0.48 mmol).
After stirred overniuht, the mixture was filtered. the filtrate was concentrated and purified by column chromatography (2% of methanol in dichloromethane as eluent) to give Compound 137 (12 mg.

7%) as white solid. II NMR (300 MHz. CDC[) 6 8.39 (s. 11-1). 7.76 (s. 111).
6.37-6.01 (m. 411).
5,41-4.78 (m. 411): 1.CMS (et) FS- 1075 (M-I-1)..
Example 20: Synthesis of Compouml 139 compound B1 0- - \i N
N CuSO4 Na2CO3 N
Me0H11-120 SM1 compound B9 1641 To a solution of compound B I (150 mg. 0.16 mmol) and SM1 (50 mg, 0.48 mmol) in Me011/1120 (4 m1/2 ml.) was added vitamine C sodium salt (63 mu. 0.32 mmol) followed with the addition of CuSO., (51 mu. 0..32 mmol) and Na:CO; (51 mifõ 0.48 mmol).
After stirred overniat, the mixture was filtered, the filtrate was concentrated and purified by column chromatography (2% of methanol in dichloromethane as eluent) to give Compound B9 (37.1 mu.
_ SUBSTITUTE SHEET (RULE 26) 22%) as white solid. III NMR (300 MI lz. CDCI3)69.37-7.89 (m. 511). 6.39-6.01 Om 411). 5.42-4,99 (m. 411): "CMS (m/i.) ES- 1040 (M-1-1)-.
Example 21: Synthesis of Compound 1111 B Si TBAF
Pd(Ph3)2C12, Cul. Et3N THE
0 80 degrees. o/n Step 2 SM1 Step 1 LOH compound B1 0 OH
-._/
Me0H OH Step 3 0 >¨N
µt\IN

compound B11 1651 To a solution of SM1 (2.1 e. 10 mmol) ill Dioxane (20 ml.) was added ethynyltrimethylsilune (2 g. 20 mmol). Cul (191 mg. 1 mmol) and I'd(PPIO:C12 (730 me. 1 mmol) under N. then 1..t,N ( 10 g. 100 mmol) was added dropw Ise. Aker stirred at 100 oC
overnieht, the mixture was quenched by water. extracted with Nt0Ac (50 mi. x 2). The combine organic layer was dried over anhydrous Na:SO4. concentrated to give crude intermediate 1.
1661 The crude intermediate I was dissolved treated with 'IBM' in THE (20 mL.. 20 mmol) at r.t. for 2 hours, then quenched by water and extracted with Et0Ac (50 ml. x 2). The combine organic layer was dried over anhydrOLIS Na:Sat. concentrated to give crude intermediate 2 which was purified bv column chromatography to eive 2 (0.9 g. 58%) as yellow solid.
III NMR (300 MHz. DM50-d6) 7.96 (d. 211). 7.62 (d. 211). 4.48 (S. 114 3.87 (s. 311).
1671 To a solution of 2 (0.5 g. 3.13 mmol) in Me011 (10 ml.) was added 1.i01-1 (0.312g. 12.52 mmol) in water (10 ml.). l'lie mixture was stirred for 2 h. then quenched by [ICI solution (2N).
extracted with Et0Ac (30 ml.*3). The combined organic was dried over anhydrous Na:S0.1.
concentrated to give the desired intermediate 3 (0,35 g, 77%) as Yellow solid.
1681 To a solution of compound 111 (130 me. 0.16 mmol) and intermediate 3 (70 me. 0.48 mmol) in Me011/11:0 (6 m1.13 mt.) was added vitamine C sodium salt (63 mg.
0.32 mmol) followed with the addition of CuSai (51 mg. 0.32 mmol) and Na:CO3 (51 me, 0.48 mmol).
After stirred overnight, the mixture was adjusted to pll about 3-4. filtered.
the filtrate was SUBSTITUTE SHEET (RULE 26) concentrated and purified b> column chromatography (1.5% of methanol in dichloromethane as eluent) to give Compound RI 1 ( 15.5 mg. 9%) as white solid. 11-1 NMR (300 MHz. CDCF)68.19 Om 3H). 7.98 (d. 21-I). 6.77-6.1 1 (m. 411). 5.49-4.51 (m. 411): 1.CMS (m/z) FS- 108; (M-1-1)-.
Example 22: Synthesis of Compound 1112 HO -Br 0 Naai Di\AF
SO,Na CH2Cl2 SO,CI
Step 1 SM1 1 Step 2 2 R
0 compound B1 11-3\2\
Step3 2NH2 Step4 O--(-SO2NH2 compound B11 1691 fo a solution of SM I 2.
23 mmol) in propan-2-ol (20 ml.) was added NaOH (2.8 g. 69 mmol) in water and 3-bromoprop-Hne (2.4 g. 20 mmol). Alter stirred for 411 at 70 C. the mixture \\ as concentrated. filtered. the filter cake was washed by water.
dried to give intermediate I (2 g. 37%) as xellow solid. 1.CMS (m/z) ES 234 (M+Na)'.
1701 bo a solution of intermediate 1 (I g. 4.3 mmol) in Mil (8 \\35 added ()vit.\ I
dichloride (1.1 g. 8.6 mmol) in DCM (4 ) dropw ise at 0 C. :1 fter stirred oµernight at Ft. the mixture \\ as quenched by \\ ater. extracted \\ ith DCM (30 ml. x 3). The combined organic was dried over anhydrous Na2SO4. concentrated. purified by column chromatography (0 to 10% of Ft0Ac in petroleum ether as cluent) to give the desired intermediate 2 (0.35 g. 77%) as yellow solid.
Intermediate 2 (0.35 g. 1.5 mmol) was added into ammonia water (5 ml.), the mixture was stirred for 1 h at r.t.. quenched by addition of water. extracted with Ft0Ac (20 ml. x 2). The combined organic \\ aS dried over an 11\ drous Na2S0.1. concentrated. pained by column chromatography (0 to 50% of Ft0Ac in petroleum ether as eluent) to gi \ e intermediate 3 (0.2 g.
20% for two steps) as yellow solid. II NMR (300 MI It. DM SO-d6) 6 7.77 (d. 211). 7.24 (s. 211).
7.14 (d. 211). 4.90 (d. 211). 3.63 (m. II I).

'1(1 a solution of compound RI ( 150 mg. 0.16 mmol) and intermediate 3 (101 mg. 0.48 mmol) in Vleoll/IFO (6 iii 3 ml.) as added µitamine C sodium salt (63 rrhl. 0.32 mmol) SUBSTITUTE SHEET (RULE 26) followed with the addition of CuS0.1 (51 mu,. 0.32 mmol) and Na2C01 (51 mu.
0.48 mmol).
After stirred overnight. the mixture \\ as filtered. and the filtrate was concentrated and purified by column chromatouraphy (0 to 2.5% of methanol in dichloromethane as eluent) to give Compound B12 (13.4 mg. 7.3%) as white solid. '11 NMR (300 MI lz. CDCI3)6 7.88 (m, 311). 7.12 (m. 211). 6.39-6.01 (m. 4H). 5.42-4.63 (m. 414): ',CMS (m/z) ES- 1148 (M-H)-.
Example 23: Synthesis of Compound 1113 Br.
Si TI3AF
Pd(Ph3)202, Cul, Et3N
TN=
80 degrees. o/n `SO2NH2 Step 2 SM1 Step 1 1 TI
.S02N Step 3 N
N

compound B13 1721 "ro a solution of SM I (2.3 g. 10 mmol) in dioxane (20 mL) was added ethynyltrimethylsilane (2 g. 20 mmol). Cu! (191 mg, I mmol) and Pd(PPh3)2C12 (730 mg. 1 mmol) under N. then Et31\1 (10 u. 100 mmol) was added drop\vis. Alter stirred overnight at I 00 C. the mixture \\ as quenched H addition of water. the mixture \\ as extracted with 11t0Ac (50 ml. x 2). The combine organic layer was dried over anhydrous Na2SO4.
concentrated to give crude intermediate I.
[731 The crude intermediate I\\ as treated w ith 'IBA I in T111. (20 nil..
20 mmol) and stirred 2h at rt.. then quenched H. water and extracted w ith 11t0Ac (50 ml. x 2). The combine organic la.\ er \\ as dried Over anhydrous Na2S01. concentrated to give crude intermediate 2.
which was purified by column chromatography to give pure intermediate 2 (1.5 g. 84%) as ,vellow solid. 1H NMR (300 MIlz. DMSO-d6) Ci 7.82 (d. 21.1). 7.68 (d. 2H).
7.46 (s. 2E1). 4.45 (s.
111).
1741 To a solution of compound 131 (150 mu. 0.16 mmol) and intermediate 2 (87 mg. 0.48 mmol) in Me011/11,0 (6 ml /3 ml.) \vas added vitamin C sodium salt (63 mu.
0.32 mmol) followed w ith the addition of CuSO4 (51 mg. 0;12 mmol). Na2CO3 (51 mu. 0.48 mmol). After stirred at r.t. overnight. the mi \t are \\ as Filtered. 'Hie Filtrate was concentrated and purified H
SUBSTITUTE SHEET (RULE 26) column chromatography (0 to 1.5% of methanol in (Iichloromethane as eluent) to give Compound B13 (48.7 mg. 27%) as white solid. II I NMR (300 MHz. (DCI3) 6 8.21 (m. II). 7.93 (m, 41I). 6.37-6.00 (m. 411). 5.44-5.30 (m. 511). 1.[MS (m/7.) NS- I 118 (M-I
Example 24: Synthesis of Compound 1314 /OH

OH
NaOH
0 compound B1 R
0 H20St 2 -N
Et0H ep HOOC
SM1 Step 1 1751 To a solution of SM1 (2.3 g. 20 mmol) in Nt01-1 (20 nil.) was added Na01-1 (1.6 g. 40 mmol) in water. then 3-bromoprop- I -yne (2.4 g. 20 mmol) was added at 0 C.
After stirred for 4h at r.t, the mixture \vas quenched by 11C1 solution (2N) and p1! was adjusted to 3-4. extracted \vith l'AOAc (50 ml *5)= The combined organic was dried over anhydrous Na2SO4.
concentrated.
purified H column chromatography (0-3% of methanol in dichloromethane as eluent) to give intermediate I (1.2 g. 40%) as colorless cr\ stal. !.CMS (m/i.) NS- 154 (NI -11)'.
1761 ft a solution of compound 131 (150 mg, 0.16 mmol) and intermediate I (73 mg. 0.48 mm(1l) in Me01-1/110 (4 iii! .2 ml,) was added vitamine C sodium salt (63 mg.
0.32 111111(11) followed \\ ith the addition of CuS0.1 (5 I mg. 0.32 mmol) and Na-,.001 (51 mg, 0.48 mmol). After stirred overnight, the mixture \vas filtered. the fl I trate was concentrated and purified by column chromatography (2% of methanol in dichloromethane as eluent) to give Compound B14 (12.6 mg. 7.2%) as white solid. 'ii NMR (300 MHz. CDCI3) 6 8.17 (m. 11-1). 6.39-5.99 Om 41-1). 5.56-4.87 (m. 411): [[MS (m//) 1/5- 1090 (M-11) .
Nnivmatic activities of rapaniNcin analogs of the present invention The m'IµOR is a serine/threonine protein kinase that has been shown to regulate multiple cellular responses including cell growth. proliferation. motility. survival and protein synthesis.
mTOR kinase activity is regulated by several upstream signaling pathways and its dysregulation has been implicated in several Corms of cancer. Now we use a l'erbillin labeled anti-phosphor lated 4N-13P I anti both to detect phosphorylation of the GI /P-labeled substrate H
mTOR. This TR-FR[]' based assay can be used to screen inhibitors of mIOR in vitro.
1781 Materials: Assay buffer components: I M IIFPES p117.5. GII3CO.
Cat#15630 I NI
MgCI:. Sigma. Cat) i MI028: 0.5M [DTA. G1BCO. CoOt 15575: DTT. Sigma Catg 43819:
SUBSTITUTE SHEET (RULE 26) EGTA. Sigma CaO( [3889: Triton X100. Sigma. Cattl 18787: BSA CALBIOCHFNI Catii 126575.
1791 Enzyme. substrate and detection reagents: mTOR: Invitrogen. Cat4 PV4753: GI2P-4L-BP I : Invitrogen. CatII PV4759: FKBP 12: SinoBiological, Catit 10268-H08E:
ATP: Sigma Cat#
A26209: 'lb-anti-1)4E-101: Invitrogen. Cat4 PV4755: TR-FRET Dilution butler Invitrogen. Cat#
PV3574.
1801 Plate: Compounds preparation plate: 384-well. Corning cat4 3657: Assay plate: Hack low volume 384\\ ell microtiter plate (Greiner Cat4 784076).
1811 Procedure: Compounds dosage gradient solution preparation:
Compounds were 3-fold serial diluted in 100% DM50 in a microtiter plate (Corning 3674) at 11 different concentrations in the range of I ()OLIN] to I .7nM (100 01. 33 0M. I
1pM. 3.7pM.
1.20M. 411 nM. 137 nM. 46 nM. 15 nM. 5 nM. 1.7 nM). Then the diluted compounds in 100%
DMSO was 10-fold diluted \\ ith ddH20. so the compounds were in 10% DMSO.
1821 A typical assay protocol of measuring the mTOR inhibitory ability of the rapamycin derivatives of the invention is as follows:
Assay protocol:
0.5 pl diluted compounds in 10% DMSO was pipetted into a black low volume 384well microtiter plate (Greiner 13io-One. Frickenhausen. Germany, ca0,' 784076):
:Vora solution of ml OR in aqueous assa \ buffer 150 mM I IEPESIXa011 pll 7.5.
5 mM
MgC12. 1 .0 infV1 clithiothreitol. 1 mM EGTA. 0.0% \/v) Triton-X100 (Sigma).
0.01 % (w/y1 bovine serum albumine ( NSA )1 (mTOR. 0.3125 nu/p > final cone. in the 5 pI
assay volume is 0.125 ng:p I) were added to the assay plate and the compound-enn me mixture was incubated for 15 min at 22'(' to allow pre-binding of the test compounds to the enzyme before the start of the kinase reaction:
1831 The kinase reaction was started by the addition of 2.5 pl of a solution of ATP (ATP. 200 pM >
final conc. in the 5 01 assa.\ volume is 10 pM) and substrate (0.8 pM ¨> final conc. in the 5 01 assa \ volume is 0.4 0M) in assay bulThr and the resulting. mixture \\ as incubated for 18 min at 22'12.
1841 The reaction \\ as stopped by the addition of Sul of 30m NI [DTA
(FDTA. 30 mM
final cone. in the 10 01 assa volume is 15mM) and 4nM Th-chelate labeled anti-4F-BP I pT461 phosphospecific ant ibod ln\ itrogen Cat!: PV47551 (lb-labeled antibock . 4 nM linal conc.
SUBSTITUTE SHEET (RULE 26) in the 10 t.t1 assay volume is 2 nM) in TR-FRn- dilution huller, the resulting mixture was incubated 1 hour at 22 C to allow the formation of complex of the phosphorylated substrate and the [b-chelate labeled antibody.
1851 l he amount of phosphorylated substrate was evaluated by measurement of the resonance energ\ transfer from the Th-chelate to the ()11). Therefore. the fluorescence emission at 495 nm and 520 nm after excitation at 340 nm was measured on envision 2104 multi label reader (Perkin-Idmer). The ratio of the emission at 520 nm and at 495 nm was taken as the measure for the amount of phosphorylated substrate. The data were normalised (enzyme reaction without inhibitor - 0 % inhibition, all other assay components but no enzyme = 100 %
inhibition) and IC50 values were calculated by a 4 parameter fit (equation (1)) using ID13S
Milt software (II) I3usiness Solutions lad.. ) Y-13ottom 1- (TOP-Bottom )/( 1 10A(( 1 .ogIC50-X )*h i 1 Islope)) equation (1) 1861 In this equation. Y was the normalized %inhibition value. X was the log value of the test compound concentration. 1050 was the concentration of compound w here half of maximal inhibition as achio.ed.
1871 The testing results of in l( inhibitory effects of the rapamycin analogs/derivatives of the invention are shown below:
Compound ID 1C50(nM) Compound ID IC50(A1) 1-5.09 Rapam! cin Compound A 1 5 6.-13 32 ! 18.43 Compound 14 ' 6604.427021 ! 133 ; 15.40 Compound A7 >10000 ' 131 28.52 Compound A 10 = >10000 135 ! 19.38 Compound 15 ! >10000 136 l2.96 Compound A6 ! 91-13.20 137 23.77 Compound A3 r:>10000 39 27.16 Compound A 13 ! >10000 ! 311 ! 446.00 SUBSTITUTE SHEET (RULE 26) Compound A 12 1 >10000 B12 35.19 Compound A 14 >10000 1313 29.03 Compound A9 - t >10000 B14 6.80 Tumor cell inhibition studies:
Cell proliferation assay 1881 The effect of different compounds on the cellular activities was quantitated through determining the number of living cells in a culture by a homogeneous detection method For quantitative determination of cell viability by the CellTiter-Glo chemiluminescence detection kit for ATP. ATP is an indicator of the metabolism of living cells.
Homogeneous detection step is added directly to the single reagent (Cell-liter-GI 'R Reagent) in serum-containing cultured cells. without washing. the cells or removing the medilffil. After adding reagent and mixing in a 96-well or 384-well plates. the number of cells that can be quantified H the system within 10 minutes. is as low as IS cells in each well.
Preparation of the reagents 1891 Different cell types \\ ere cultured using a medium. containing. 10%
1:13S plus I%
penicillin streptomycin double antibiotics, and the follow ing appropriate additives: DMIAI
medium (Cibco. Item No. I 19)5073) for culturing colorectal cancer cells I ICT
I 16. breast cancer cells MCF-7 and MDA-MB-231 melanoma cells SK-MEL-28. A549 and epidermal squamous cell carcinoma cell A431: RPM 1-1640 medium (containing 2 niM L-glutamine. 1.5 g / 1. sodium bicarbonate. 4.5 u / I glucose. 10 RIM 11EITS. 1.0 mM sodium pyruvate ( Item #1 72400-120 from Giber)) fOr culturing r87/VIG and kidney caneer786-0: HI-2K mixed medium (Item #21127 From Gibe()) For culturing prostate cancer cell line PC-3, Instrumentation Multi-label Micro-plate Reader rnvison 21-1 From Perkin rimer Cell Culture conditions 1901 Al! 9 cell lines were cultured in the wells in the plates at a cell density of 3000 cells/well alter 9 passages.
Preparation or culture media and cell culture conditions:
SUBSTITUTE SHEET (RULE 26) Prepare the compounds and condition the cells the next day: each chemical compound to be aSSaNed was diluted to 10 niM stock solution with 100% DM SO. followed by additional dilution with 100% DiX150 diluted to 2 mN1. followed b\ serial 5X dilution using serum-free cell culture medium to a final 10 di fferent diluted concentrations points (2000.
400. 80. 16, 3.2. 0.64.
0.128, 0.0256, 0.00512. 0.00102 [AM). plus 0.5% {WS (no compound) as a maximum control and 10WV1 Rapamycin as a minimum control. A solution of 0.5til of each diluted compound is added to the 100W 0i-cell culture plate. the final compound concentration of 10 points (10. 1 0.4.
0.08. 0,016. 0.0032. 0.00064. 0.000128. 0.0000256. .00000512 [AM). '11he cells were then cultured in 37 'V incubator for 72 hours. In order to ensure die reliability of the experiments of determining. the inhibition of each compound. a duplicate was used for each compound concentration gradient w ill do two repeated (lable 1). and the determination of each compound was repeated tw ice.
Plate reading After 72-hr of cell culture. 50 ul of CellTiter Glo was added to each well on the plate. and shaken for 5 min on a shaker followed by 10 min at room temperature. The cell number was analyzed by the Micro-Plate reader.
Data analysis:

Cell viability was obtained through the reading by the multi-label micro-plate reader. The effect of dilution value on the % as calculated using the follow ing fomula:
% cell inhibition- 100-100, I Signal-low control )/( High control-low control.
in which signal low control. and high control are the test compound. minimal value. and maximal value respectively.
[941 The 1050 value of each test compound in inhibiting the cells is obtained by formula 2 (below ):
Bottom OP-liottom rt 1 0 lC50/X hillslopco. in w hich X and v are known values. IC50.
1 lillslope. Top and I3ottom 4 parameters generated by the analysis software.
V as the %
inhibition. X as the test compound concentration, and IC50 as the concentration of the compound needed to inhbit 50% of the cells. 1 lillslope is the slope of curve fitting.
usually around I.
1951 All experimental data w Cie analyzed by IDI3S X1.111.5 (II) 13LISinCSS
Solutions ).
Experimental results and conclusion All the potency of each test compound is shown in one of the Hlow kW, graphs for each of the cancer cell models tested. The lower the ellrve in the graph. the more potent each SUBSTITUTE SHEET (RULE 26) compound is. From the data shown in each the graphs. it is clear that all the series 13 compounds showed varying levels of high potency against the cancer cell tested. Some of the 13 series compounds were extremely potent. reaching a potency level of nM
concentration range.
Renal cell carcinoma tumor cell inhibition studies:
Renal cell carcinoma tumor cell in studies: Figl and Fig2.
Lung Cancer A549 cell inhibition studies: Fig3, Eig4 and Fig5.
Melanoma SK-MEL-28 cell in studies: Eig6, Fig7 and Fig8.
Epidermal cancer A431 tumor cell model: Fig9, Eig10 and Fig11.
Glioblastoma 1187 MG Tumor model studies: Fig12, Fig13 and Fig14.
Human colorectal tumor HCT 116 model studies: Fig15, Fig16 and Fig17.
Breast cancer MDA-MB-231 tumor model: Fig18, Fig19 and Fig20.
Breast cancer MC7F-7 tumor model: Fig21, Fii,422 and Fig23.
Prostate cancer PC-3 tumor studies: Fig24, Fig25 and Fig26.
Efficacy studies of raparilvein derivatives in Human Colon Tumor (11CT116) model Purpose: The objecti\ e or this stud\ is to evaluate preclinicallv the in vivo therapeutic ellicac\ 011,:\ IS (positive control) and a lead compound from the 13 series administrated as per os (p.o.) in the slowing or eliminating tumor development in subcutaneous I IC F-116 human colon cancer model.

Animals: 13a1b/c nude mice. female. 6-8 weeks. \vcighinu approximately 18-20g.
A total or 70 will be needed For the study. which µvill be purchased From Vital River Laboratory Animal Technology Co. ltd.

Tumor Inoculation: 1:.ach mouse will be inoculated subcutaneously at the right [lank with IR-1-116 tumor cells 0\10() in O. I ml or P1115 For tumor development. The treatments w ill be started when the tumor size reaches approximately -150 mm3. The test article administration and the animal numbers in each group are shown in the Follow im2 experiment design table, Groups and Treatments 1 Dose Dosing )osi 7 Group 11 I1.Catillt2nt Schedule (mg/kg) Rout volume Vehicle p.o. 10 pl/v, QD x21 10 A 5 9 p.o. I 0 W/g QD x 21 SUBSTITUTE SHEET (RULE 26) j 3 10 B p.o. I 0 1.11/g QD x 21 4 10 B 9 p.o. 10 tAl/g QD x 21 5 10 18 p.o. 10 1.1.1/g QD x 21 Note: n: animal numher: Dosing volume: adjust dosing volume based on body weight 10 pi/g), Treatment schedule may be adjusted i hocky \\ eight loss > 15%.
11001 Assignment to Groups: Before commencement of treatment. all animals will be weighed and the tumor volumes will he measured. Since the tumor vollillle can affect the effectiveness of any given treatment. mice \\ ill he assigned into groups using randomized block design based upon their tumor kolumes, This ensures that all the groups are comparable at the baseline, 11011 1:.ndpoints: The major endpoint is to see if the tumor growth can he delayed or mice can be cured. I umor sites will he measured twice weekly in two dimensions using a caliper. and the volume will he expressed in mm3 using the formula: V --- 0.5 a x b2 where a and h are the long and short diameters or the tumor. respectively. The tumor sizes are then used for the calculations or both T-C and T/C values. T-C is calculated with T as the median time (in days) required for the treatment group tumors to reach a predetermined site (e.g.. 500 mm3). and C is the median time (in days) for the control group tumors to reach the same site. 'file T/C
value (in percent) is an indication of antitumor effectiveness. T and C are the mean volume of the treated and control groups. respectivek. on a gi\ en (LIV. lumor tiStilleti will he collected 16r the tumor weight and photo at the end of the stud\ .
11021 Termination:Th I is stuck. \\ ill he terminated when the mean tumor size of the control group reach the volume of 600- 1000 mm3. Animals that are observed to he in a continuing deteriorating condition \\ ill be euthanized prior to death, or before reaching a comatose state.
Animals showing obvious signs of severe distress and/or pain should he humanely sacrificed. In case of following situations. the animals will be euthanized:
11031 Animals have lost significant body mass (emaciated). Obvious body weight loss > 2094):
11041 Animals cannot get to adequate I6od or water.
11051 the study \\ ill he terminated \\ ith all animals in all groups being sacrificed \\ hen the mean tumor burden in the \ chicle treated control group reaches a value of 2000 mm3.
11061 Statistical Analysis: I=or comparison between two groups. an independent sample t-test \\ill he used. For comparison among three or more groups. a one-\\ a.) ANOVA
will he SUBSTITUTE SHEET (RULE 26) performed. II a signilicant I -statistics (a ratio of treatment variance to the error variance) is obtained, multiple comparison procedures will be applied after ANOVA. The potential synergistic effect between treatments will he anakted by LSD or Dunnett's 13.
All data will he anal \ zed using, I 7.() solk\ are. p 0.0 is considered to he statisticall significant.
11071 Summar\.: As shown in the follo\\ ng figure. after 22 days of treatment.
the 9 mg/kg/day of A I 5 positive control compound (A In tor from Novartis) significantly inhibited tumor growth by 63% in Sub() LICT I 16 resistant colon cancer xenograft model(* P<0.05), which is consistent with the reports from literature. After 22 days treatment. 11 compound suppressed the tumor growth by 42%. 57% and 64% (**. P<0.01). at 3. 9. and 18 mg/kg/day, respectively. compared to the vehicle control. No obvious toxicity was observed, These data indicate that B compound is very potent in vivo and ma\ O\ ercome the colon cancer resistance.
Fig 27. In this figure. the top line (diamond) is lot- Vehicle, second line (triangle) is For 137 at 3 mg /Kg. dose: third line (purple cross) represents 137 at 9gm/Kg dose: rourth line from the top (pink square() represent llini(or at 9 mg/Kg dose). the bottom line (blue cross) represents 137 a I 8 mg: Kg dose.
Methods offreatment 11081 The compounds of the present invention. including but not limited to those specified in the examples, possess immunomodulatory and anti-tumor activity in mammals (especially humans). As immunosuppressants. the compounds of the present invention are useful for the treatment and prevention of immune-mediated diseases such as the resistance by transplantation or organs or tissue such as heart. kidney. liY=er. medulla ossill111. skin.
cornea. lung. pancreas.
intestinum ten ue. limb. muscle, nerves. duodenum, small-bowel. pancreatic-islet-cell, and the like: \ ersus-host diseases brought about b\ medulla ossium transplantation: autoimmune diseases such as rheumatoid arthritis. s\ stemic lupus er\ theinatosus.
Hashimoto's th multiple sclerosis. myasthenia gravis. type I diabetes. uveitis. allergic encephalomyelitis.
glomerulonephritis. and the like. Further uses include the treatment and prophylaxis ols intlammator and hyperprolilerative skin diseases and cutaneous manifestations of immunologically-mediated illnesses. such as psoriasis. atopic dermatitis, contact dermatitis and further eczematous dermatitises. is seborrhoeis dermatitis. lichen plaints.
pemph4n.ts, bulious pemphigold. epidermic)] \ sis buliosa. urticaria. angioedemas. vasculitides.
ervthemas. cutaneous SUBSTITUTE SHEET (RULE 26) cosinophijias. lupus erythematosus, acne and alopecia areata: various eye diseases (autoimmune and otherwise) such as keratoconjunctivitis, vernal conjunctivitis. aveitis associated with Behcers disease. keratitis. herpetic keratitis. conical cornea. dystrophia epithelialis corneae.
corneal leukoma. and Dellini pcmphigus. In addition. reversiHe obstructive airway disease.
which includes conditions 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 alma hyper-responsk cuss). bronchitis, allergic rhinitis. and the like are lar,(4eteci b\ compounds of the present invention. In of mucosa and blood vessels such as !.astric ulcers. vascular damage caused lw isehemic diseases and thrombosis. Moreover.
hyperproliferative vascular diseases such as intimal smooth muscle cell hyperplasia. restenosis and vascular occlusion. particularly following biologically- or mechanically-mediated vascular injury. may be treated or prevented by the compounds of the present invention.
Other treatable conditions include ischemic bowel diseases. inflammatory bowel diseases.
necroti/ing enterocolitis. intestinal in such as Coeliac diseases. proctitis, eosinophilic gastroenteritis. mastoevtosis. Crohn's disease and ulcerative colitis: nervous diseases such as multiple m ositis. Guillain-Barre syndrome. Meniere's disease. polyneuritis.
multiple neuritis.
mononeuritis and radiculopath\ : endocrine diseases such as 11\ perthyroidism and Rasedow's disease: hematie diseases such as pure red cell aplasia. aplastic anemia.
hypoplastic anemia.
idiopathic thrombocytopenie parpura. autoimmune hemolytic anemia.
agranuloeytosis.
pernicious anemia. megaloblastic anemia and anerythroplosia: 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.
photoallergie sensitivity and cutaneous T cell lymphoma: circulatory diseases such as arteriosclerosis, atherosclerosis. aortitis syndrome. polyarteritis nodosa and myoeardosis:
collagen diseases such as seleroderma. Wegener's ._?,ranulonta and Siouren's syndrome: adiposis:
eosinophilic fasciitis: periodontal disease such as lesions of gingiva.
perioclontium. alveolar bone and substantia ossea dent is: nephrotic syndrome such as glomerulonephritis:
male pattern alCONCIL1 or alopecia senilis b\ preventinu epilation or pros di in hair germination and'or promoting hair generation and hair growth: muscular d stroph\ : l'voderma and Setar\
s\ ndmme: Addison's disease: aeti\ e oxygen-mediated diseases. as for example organ injUry sLich as ischemiLi-reperlusion injur\ 01' organs (such as heart. liver. kidney and digestive tract) which SUBSTITUTE SHEET (RULE 26) occurs upon preservation. transplantation or ischeme disease (for example.
thrombosis and cardiac infarction): 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 bleonkeins). lung cancer and pulmonary emphysema:
ocular diseases such as cataracta. siderosis. retinitis, pigmentosa. senile macular degeneration. vitreal scarring and conical alkali burn: dermatitis such as erythema multiforme. linear lgA
ballous dermatitis and cement dermatitis: and others such as gingivitis. periodontitis. sepsis.
pancreatitis. diseases caused b\. environmental pollution (for example, air pollution). aging.
carcinogenesis. metastasis of carcinoma and hypobaropathy: diseases caused by histamine or leukotriene-C.subA release:
fichcers disease such as intestinal-. yasculo- or neuro-Behcers disease. and also Behcet's which affects the oral cavik. skin. tne. vulva, articulation. epidick mis. lung.
kidney and so on.
urthermore. the compounds of the present invention ma\ be USCILI I or the treatment and prevention of hepatic disease such as immunogenic diseases (for example.
chronic autoimmune liver diseases such as autoimninune hepatitis. primary biliarv cirrhosis and sclerosing cholangitis). partial liver resection, acute liver necrosis (e.g. 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-on-chronic- liver failure (acute liyer failure on chronic liver diseases). and moreover are useful for various diseases because of their usefal act k it such as augniention of chemotherapeutic effect. evtomegalovirus infection. particularl I
ICVIV infection. anti-inflammatory activik, sclerosing and fibrotic diseases such as nephrosis.
scleroderma.
pulmonary fibrosis. arteriosclerosis. congestke heart Failure. ventricular h\
pertrophy. post-surgical adhesions and scarring. stroke. myocardial infarction and illjUry associated with ischemia and reperfusion. and the like.
11091 Additionally. compounds of the present invention possess rk-506 antagonistic properties.
The compounds of the present invention may thus be used in the treatment of immunodepression or a disorder involving immunodepression. rxamples of disorders involving immunodepression include cancer. fungal infections. senile dementia. trauma (including wound healing.
surgery and shock) chronic bacterial infection, and certain central nervous system disorders. The immunodepression to be treated may be caused by an overdose of an immunosuppresske SUBSTITUTE SHEET (RULE 26) macrocyclic corn pound.
for example derivatives of 1 2-(2-cyclohexv1- 1 -methylvi ny1)-1 3.1 9.2 1.27-tetramethyl- 1 1 .28-dioxa4-azatricyclo122.3. 1 .0⁴.91octacos- 1 8-cue such as 1:1;,-506 or rapamycin. The overdosing of such medicants by patients is quite common upon their realizing that they have forgotten to take their medication at the prescribed time and may lead to serious side effects.
11101 The compounds of the present invention, including but not limited to those specified in the examples. possess anti-tumor activity in mammals (especially humans). As an anti-cancer drug, the compounds of the in\ cut ion can be used to treat brain and neurovascular tumors, head and neck cancers, breast cancer. lung cancer. mesothelioma. lymphoid cancer, stomach cancer.
kidney cancer. renal carcinoma, liver cancer and liver cirrhosis, ovarian cancer, ovary endometriosis. testicular cancer, skin cancer. melanoma, neuro and all endocrine cancers, spleen cancers. pancreatic cancers. blood proliferative disorders such as Hodgkin's cancer. lymphoma.
leukemia, and all \ cancer disorders that result from uncontrolled cellular proliferations.

Hie COM pounds of the present invention. alaV be mixed \\ itil COM1110111 V
known pharmaceutical excipients such as rfudragit. sodium carboxymethylcellulose (Na CM). sodium carboxyprop\ !cellulose. an\ other naturally derived or s\ IlthetiC C el pients to effect an efficacious pharmaceutical hormulation. The formulation comprising the compounds of the invention Illa \ be made as a immediate release formulation. or a sustained release formulation. or site injection depot formulation. depending on the medical needs. The compound of the present invention may also be combined with a medical device, such as a stent. a balloon, a balloon catheter. an orthopedic device, to further enhance the efficacy of the medical device. The compound of the present invention may he the main 1Mnction component of a medical treatment regime. such as a local injection formulation. or an ancillary function. such as a coating on a medical de\ ice. or in combination with a low-molecular weight or polymer excipient. and used as a coating or filler of' a medical device.
11121 When used to treat restenosis following a balloon angioplasty or stent placement. the compounds of the present in and the native rapamycin. are thought to exhibit their therapeutic functions through the inhibition of the mammalian target of rapamycin or mTOR.
he ma \ also bind to 1:1:131' receptors.
11131 When used in the abo\ c or other treatments. a therapeutically effective amount of one of the compounds of' the present invention may be employed in pure form or. where such forms SUBSTITUTE SHEET (RULE 26) exist. in pharmaceutically acceptable salt. ester or prod rug form..Alternatelv. the compound rria be administered as a pharmaceutical composition containing the cornpound of interest in combination with one or more pharmaceutically acceptable excipients. The phrase "therapeutically effective amount- of the compound of the present invention means a sufficient amount of the compound to treat disorders. at a reasonable benefit/risk ratio applicable to any medical treatment. It will be understood. however. that the total daily isae of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. the specific therapeutically effective dose level for any particular patient \\ ill depend upon a varlet\ of ractors includ* the disorder being treated and the se\ erit\ of the disorder: activit\ or the specilic compound emplo.\,ed: the specific composition employed: the age. body \\ eight. general health. sex and diet of the patient: the time of administration. route of administration. and rate or excretion of the specific compound employed: the duration of the treatment: drugs used in combination or coincidental 1\ ith the specific compound employed: and like factors well known in the medical arts.
For example. it is well within the skill of the art to start doses of the compound at levels lower than required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved.

Ihe total dank dose of the compounds of the present in\ ention administered to a human or lower mammal may rant:e from about 0.01 to about 20 mg/kg/day. I or purposes of oral administration, more prererable doses may be in the range of from about 0.001 to about 3 mg/4.)./da\ . lfdesircd. the effective daily dose ma\ be divided into multiple doses for purposes of administration: consequentk. single dose compositions ma\ contain such amounts or submultiples thereof to make up the (Jail\ dose. 1 opical administration may involve doses ranging from 0.001 to 10 percent mg/kg/clay, depending on the site of application. When administered locally to treat restenosis and vulnerable plaque. the dose may range from about I
microgram:mm stent length to about 100 microgram/mm stem length.
Pharmaceutical Compositions 11151 The pharmaceutical compositions of the present invention comprise a compound and a pharmaceuticalk acceptable carrier or e\cipient. which mav be administered orally. rectalk.
parentera II\ i ntrac isterna ntrayagi na I\
intraperitonealr\ . topical I\ (as b\ pow ders.
ointments, drops or transdermal patch). bucalk. or as an oral or nasal spray.
The phrase SUBSTITUTE SHEET (RULE 26) -pharmaceutically acceptable carrier- means a non-toxic solid. semi-solid or liquid filler. diluent.
encapsulating material or Formulation auxiliar\ of any type. The term -parcnteral,- as used herein. refers to modes of administration which include intravenous.
intramuscular.
intraperitoneal. intrasternal. subcutaneous and intraarticular injection and infusion.
11161 Pharmaceutical compositions of the present invention for parenteral injection comprise pharmaceutical l> acceptable sterile aqueous or nonaqueous solutions.
dispersions. suspensions or emulsions as well as sterile powders iqr reconstitution into sterile injectable solutions or dispersions just prior to use. kxamples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol.
propylene glycol.
polyethylene glycol. and the like), carboxymethylcellulose and Suitable mixtures thereof*.
vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate. Proper fluidit\ may be maintained, lqr example. bv the use of coating materials such as lecithin, by the maintenance of the required particle sin in the case of dispersions. and by the use of surfactants.
11171 I hese compositions ma\ also contain adju \ ants such as preser\
atives. wetting agents.
emulsifying agents. and dispersing agents. Prevention of the action of microorganisms ma\ be ensured H the inclusion of various antibacterial and antilnngal agents. for example, paraben.
chlorobutanol. phenol sorbic acid. and the like. It may also be desirable to include isotonic agents such as stwars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.
11181 In some cases. in order to prolong the effect of the drug. it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This inay be accomplished by the use of a liquid suspension of crystalline or amorphous material \\ ith poor water solubility.
The rate of absorption or the drug then depends upon its rate of dissolution which. in turn. inay depend upon cr\ stal siie and cr\ stall inc form. Alternatek.. delayed absorption of a parenterall.\
administered drug lqrm is accomplished H dissolving or suspending the drug in an oil vehicle.
11191 Injectable depot lqrms arc made H. Forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release may he controlled. kxamplcs of other biodegradable polymers include poly(orthoesters) and SUBSTITUTE SHEET (RULE 26) poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.
11201 The injectable Ibrmulations may be sterilized, for example. by filtration through a bacterial-retaining, illter, or bv incorporating sterilizing agents in the form of sterile solid compositions which mav be dissolved or dispersed in sterile water or other sterile injectable medium just prior to use.
11211 Solid dosage forms or oral administration include capsules. tablets, pills. powders. and L2,l'atillles. In such solid dosage forms. the acti \ e compound is mixed with at least one inert.
pharmaceuticall \ acceptable excipient or carrier such as soditim citrate or dicalcium phosphate and/or a) fillers or extenders such as starches. lactose. sucrose. glucose.
mannitol. and silieic acid.
h) binders such as carboxymethvIcellulose. alginates, gelatin.
polyvinylpyrrolidone. sucrose, and acacia. c) humectants such as glycerol. d) disintegrating agents such as agar-agar, calcium carbonate. potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin. r) absorption accelerators such as quaternary ammonium colnpounds.
welli119, agents such as cetvl alcohol and glycerol monostearate. h) absorbents such as kaolin and bentonite clay. and i) lubricants such as talc.
calcium stearate.
magnesium stearate. solid polyethylene glycols. sodillin laUrVI sulfate. and mixtures thereof. In the case of capsules. tablets and pills, the dosage form ma\ also comprise buffering agents.

oIid compositions or a similar type ma \ also be employed as fillers in soft.
semi-solid and hard-tilled gelatin capsules or liquid-filled capsules using such excipients as lactose or milk sugar as well as high molecular wcii2ht pol \ ethylene i.11\ cols and the like.
11231 The solid dosage forms of tablets. dragees. capsules. pills. and granules may he prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain ()pacifying agents and may also he or a composition that they release the active ingredient(s) only. or preferentially. in a certain part of the intestinal tract. optionall\ . in a dela\ ed manner. Lxamples of embedding compositions \vhich lila\ he used include pol\ meric substances and waxes.

lite active compounds ma\ also be in a micro-encapsulated form. i I
'appropriate. with one or more of the above-mentioned cxcipients.
11251 Liquid dosage forms for oral administration include pharmaceutietill acceptable emulsions. solutions, suspensions. syrups and elixirs. In addition to the active compounds. the SUBSTITUTE SHEET (RULE 26) (lU rd dosa!:,,c forms may contain inert chluents commonly used in the art, for example. \\ ater Or other solvents. solubili/ing agents and emulsifiers such as eth \ I alcohol.
isopropyl alcohol. ethyl carbonate. eth I acetate, ben/y1 alcohol. benn bentoate. propylene glycol. 1.3-butvlene dirneth Ibrmamide, oils (in particular. cottonseed. groundnut. corn, germ. olive.
castor, and sesame oils). glycerol. tetrahydrofurfuryl alcohol. polyethylene glycols and fattv acid esters of sorb tan, and mixtUreti there01.
11261 Besides inert diluents. the oral compositions may also include adjuvants such as wetting agents. emulsifying and suspending agents. s\\ eetening. flavoring, and perfuming agents.
11271 Suspensions. in addition to the active compounds. may contain suspending agents. for example. ethox\ kited isostear\ I alcohols, polvox \ ethylene sorbitol and sorbitan esters.
microcrystalline cellulose. at metahydroxide. bentonite. agar¨agar. and tragacanth. and mixtures thereof.
11281 Topical administration includes administration to the skin or mucosa.
including surfaces of the lung and e\ e. Compositions for topical administration, including those for inhalation. may be prepared as a dry powder which may he pressurized or non-pressuri/cd. In non-pressurized powder compositions. the active ingredient in finely divided form may he used in admixture with a larger-sized pharmaceutically acceptable inert carrier comprising particles having a size, for example. of up to 100 micrometers in diameter. Suitable inert carriers include sugars such as lactose. Desirabk. at least 95 percent by weight of the particles of the active ingredient have an effective particle site in the range of 0.01 to 10 micrometers. Compositions for topical use on the skin also include ointments. creams. lotions. and gels.
11291 Alternatek. the composition ma\ be pressuri/ed and contain a compressed gas. such as nitrogen or a I killerlCd gas propellant, The liquelied propellant medium and indeed the total composition is preferably such that the active ingredient does not dissolve therein to any substantial extent. The pressuriyed composition Ma\ also contain a surface active agent. The surface active agent ma\ be a liquid or solid non-ionic surface active agent or may be a solid anionic surface active agent. It is preferred to use the solid anionic surface active agent in the form of a sodium salt.
11301 A further form of topical administration is to the eye, as for the treatment of immune-mediated conditions of the c\e such as autoimmunc diseases. allergic or inflammatory conditions, and corneal transplants. The compound of the present in is deli ered in a SUBSTITUTE SHEET (RULE 26) pharmaceutically acceptable ophthalmic vehicle. such that the compound is maintained in contact \\ ith the ocular surface for a sufficient time period to allow the compound to penetrate the corneal and internal regions of the e\ e. as for example the anterior chamber. posterior chamber. \ itreous bock. aqueous humor. \iitreous humor, cornea. iris/eilar.
lens. choroidiretina and sclera. The pharmaceuticall \ acceptable ophthalmic vehicle may. for example. be an ointment. \ egetablc oil or an encapsulatinu material.
[1311 Compositions for rectal or vaginal administration are preferobl \
suppositories or retention enemas which may be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers such as cocoa butter. pol, ethylene glycol or a suppository wax which are solid at room temperature but liquid at bed.) temperature and therefbre melt in the rectum or \ aginal cavity and release the active compound.
11321 Compounds of the present invention may also be administered in the form of liposomes.
As is known in the art. liposomes are general IN derived from phospholipids or other lipid substances. I iposomes are formed b\ mono- or multi-lamellar Indrated I iquid crstals that are dispersed in an aqueous medium. Any non-toxic. physiological l\ acceptable and mctabolitable lipid capable of 1brming liposomes can be used. The present compositions in liposome form ma.
contain. in addition to a compound of the present in\ cntion. stabiliiers.
preservatives. excipients.
and the like. The preferred lipids ore the phospholipids and the phosphatid cholines (lecithins).
both natural and ti\ ntlICliC. Methods to form liposomes are known in the art.
See. for example.
Prescott. rid.. Methods in Cell Biology. Volume XIV. Academic Press. New York.
N.Y. (1976).
p. 33 et seq.
11331 Compounds of the present invention may also be coadministered with one or more immunosuppressant agents. I he immunosuppressant agents w ithin the scope of the present invention include IM1 azathioprine sodium. brequinar sodium. SPANIDIN.RTNI.
gusperimus trihvdrochloride (also known as deoxyspergualin). mizoribine (also known as brechnin). CH 1 ChiPT.RIAl. mvcophenolate moletil.
Cvlosporin A (also marketed as di flerent formulation of L\ eloTorin \
under the trademark PROGIZALIZ VI. tocrolimus (also known as K-506). sirolimus and RAPAMLN
leflunomide (also known as Il\\ A-186). glucocorticoids. such as prednisolone and its clerk-A \ es. antibody therapies such as orthoelone (OK
13) and lenopax.R I Ni.. and antith\ m oc te globulins, such as thYmoglobulins.
SUBSTITUTE SHEET (RULE 26) 11341 'Ihe local deliver of drug/drug combinations from a stem or other implantable device has the follow ino advantages: namek. the pre \ ention of vessel recoil and remodeling through the scaffolding action of the stem and the prevention of multiple components of neointimal hyperplasia or restenosis as well as a reduction in inflammation and thrombosis. This local administration of drugs, agents or compounds to stented coronary arteries may also have additional therapeutic benefit. For example. higher tissue concentrations of the drugs. agents or compounds ma\ be achic\ ed utilizing local deli\ en. rather than s\ stemic administration. In addition, reduced systemic toxicit\ ma\ be achieved utilizing local deliver\
rather than s.stemic administration while maintaining higher tissue concentrations. Also in utilizing local delivery from a stem rather than systemic administration. a sin,,de procedure ma\
suffice with better patient compliance. An additional benefit of combination drug. agent. and/or compound theram ma\ be to reduce the dose of each of the therapeutic drugs. agents or compounds. thereb limiting their toxicit\ . while still achic\ Ma a reduction in restenosis.
inflammation and thrombosis. Local stent-based therapy is therefore a means of improving the therapeutic ratio tellicac.0oxicity) of anti-restenosis. anti-illflammator.). antithrombotic drugs. agents or compounds.
11351 It is understood that the foregoing detailed description and accompanying examples are merel illustrati \ e and are not to be taken as limitations upon the scope of the invention. which is defined solek b\ the appended claims and their cqui\ alents. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art.
Such changes and modifications. including \\ ithout limitation those relating to the chemical structures. substituents.
deri\,ati\.es. intermediates. s\ nthescs. formulations andlor methods of use of the invention. may he made \\ ithout departing from the spirit and scope thereof.
SUBSTITUTE SHEET (RULE 26)

Claims

1. A compound of Formula I or a pharmaceutically acceptable salt or prodrug thereof:
wherein A is selected from the group consisting of:
a) hydrogen, alkyl and substituted alkyl, alkenyl and substituted alkenyl, alkynyl and substituted alkynyl, cycloalkyl and substituted cycloalkyl, heterocycloalkyl and substituted heterocycloalkyl; the substitution group including hydroxyl, sulfonyl, carbonyl, amino, cyano, halogen, alkoxy, aryl, and heteroaryl, and b) aryl and substituted aryl, heteroaryl and substituted heteroaryl; the substitution group including hydroxyl, halogen, amino, carbonyl, cyano, nitro, sulfonyl, alkyl, alkoxy, cycloalkyl, heterocycloalkyl.

2. The compound from claim 1, wherein the compound is selected from the group consisting of:

3. A compound of Formula II or a pharmaceutically acceptable salt or prodrug thereof:
wherein B is selected from the group consisting of:
a) hydrogen, alkyl and substituted alkyl, alkenyl and substituted alkenyl, alkynyl and substituted alkynyl, cycloalkyl and substituted cycloalkyl, heterocycloalkyl and substituted heterocycloalkyl; wherein each substituent is independently hydroxyl, sulfonyl, carbonyl, amino, cyano, halogen, alkoxy, aryl, or heteroaryl, and b) aryl and substituted aryl, heteroaryl and substituted heteroaryl; wherein each substituent is hydroxyl, halogen, amino, carbonyl,cyano, nitro, sulfonyl, alkyl, alkoxy, cycloalkyl, or heterocycloalkyl.

4. The compound from claim 3, wherein the compound is selected from the group consisting of:

5. A pharmaceutical composition comprising a compound of any of claims 1-4 and a pharmaceutical excipient.

6. The pharmaceutical formulation of claim 5, wherein the formulation is suitable for administration to a mammal via a route selected from the group consisting of oral, nasal, intravenous, transdermal, parenteral, subcutaneous, intramuscular, intra-ocular, and peritoneal routes.

7. The pharmaceutical formulation of claim 6, wherein the mammal is a human.

8. A method of treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any of claims 1-4.

9. The method of claim 8, wherein the cancer is selected from the group consisting of brain and neurovascular tumors, head and neck cancers, breast cancer, lung cancer, mesothelioma, lymphoid cancer, stomach cancer, kidney cancer, renal carcinoma, liver cancer and liver cirrhosis, ovarian cancer, ovary endometriosis, testicular cancer, skin cancer, melanoma, neuro and all endocrine cancers, spleen cancers, pancreatic cancers, blood proliferative disorders such as Hodgkin's cancer, lymphoma, leukemia, and any cancer disorders that result from uncontrolled cellular proliferations

10. A method for treating or preventing an immune-mediated disease in a subject, comprising administering to the subject in need thereof a therapeutically effective amount of a compound of any of claims 1-4.

11. The method of claim 10, wherein the immune-mediated disease is selected from the group consisting of resistance by transplantation of heart, kidney, liver, medulla ossium, skin, cornea, lung, pancreas, intestinum tenue, limb, muscle, nerves, duodenum, small-bowel, or pancreatic-islet-cell; graft-versus-host diseases brought about by medulla ossium transplantation.

12. The method of claim 10, wherein the immune-mediated disease is rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes, uveitis, allergic encephalomyelitis, or glomerulonephritis.

13. The method of claim 10, wherein the immune-mediated disease is a graft-versus-host disease brought about by medulla ossium transplantation.

14. Use of a compound of any of claims 1-4 for manufacturing of a medicament for the treatment of a cancer or an immune-mediated disease.

15. The use of claim 14, wherein the cancer is selected from the group consisting of brain and neurovascular tumors, head and neck cancers, breast cancer, lung cancer, mesothelioma, lymphoid cancer, stomach cancer, kidney cancer, renal carcinoma, liver cancer and liver cirrhosis, ovarian cancer, ovary endometriosis, testicular cancer, skin cancer, melanoma, neuro and all endocrine cancers, spleen cancers, pancreatic cancers, blood proliferative disorders such as Hodgkin's cancer, lymphoma, leukemia, and any cancer disorders that result from uncontrolled cellular proliferations; and the immune-mediated disease is selected from the group consisting of resistance by transplantation of heart, kidney, liver, medulla ossium, skin, cornea, lung, pancreas, intestinum tenue, limb, muscle, nerves, duodenum, small-bowel, or pancreatic-islet-cell; graft-versus-host diseases brought about by medulla ossium transplantation.