Cyclic dinucleotide compounds and methods of use

Abstract

Disclosed are cyclic dinucleotide compounds and methods of use, methods for synthesizing the compounds, pharmaceutical compositions containing the compounds, and uses of the compounds and compositions in medical treatment.

Description

[0001] This application is a divisional application of a patent application with application number 201780030376.9, application date March 17, 2017, and invention name “Cyclic dinucleotide compounds and methods of use”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Application No. 62 / 310,364, filed on March 18, 2016, U.S. Provisional Application No. 62 / 355,382, filed on June 28, 2016, and U.S. Provisional Application No. 62 / 396,140, ​​filed on September 17, 2016, the entire contents of each of which are incorporated herein by reference. Technical Field

[0004] The present invention provides novel cyclic dinucleotide cGAMP analogs, pharmaceutical compositions thereof, methods for their synthesis, and their use in medical therapy. In particular, the compounds of the present invention enhance the body's immune response by activating STING (stimulator of interferon genes), and can be used for immunotherapy of cancer, infectious diseases, and immune disorders. The compounds can also be used as adjuvants for the development of vaccines against cancer and infectious diseases. Background Art

[0005] Cytosolic DNA induces type I interferons and other cytokines, which are important for immune defense against microbial infection and malignant cells, but can also lead to autoimmunity. This DNA signaling pathway requires the adaptor protein STING (stimulator of interferon genes) and the transcription factor IRF3, but the DNA sensing mechanism was unclear until recently. WO 2014099824 from the University of Texas discloses that mammalian cytosolic extracts synthesize cyclic-GMP-AMP (cGAMP) in vitro from ATP and GTP in the presence of DNA but not RNA. DNA transfection or DNA virus infection of mammalian cells also triggers cGAMP production. cGAMP binds to STING, resulting in activation of IRF3 and induction of type I interferons, including beta interferon (IFN-β). Therefore, cGAMP represents the first cyclic dinucleotide in complex cells, and its function is to trigger interferon production in response to cytosolic DNA as an endogenous second messenger.

[0006] By biochemical fractionation and quantitative mass spectrometry, the inventors of WO 2014099824 also identified a cGAMP synthase (cGAS) that belongs to the nucleotidyl transferase family. Overexpression of cGAS activates the transcription factor IRF3 and induces IFN in a STING-dependent manner. Knockdown of cGAS inhibits IRF3 activation and IFN induction by DNA transfection or DNA virus infection. cGAS binds to DNA in the cytoplasm and catalyzes cGAMP synthesis. These results indicate that cGAS is a cytosolic DNA sensor that induces interferon by producing the second messenger cGAMP. The inventors of WO 2014099824 also determined that the second messenger cGAMP they isolated and synthesized contained two phosphodiester bonds, one between the 2'-OH of GMP and the 5'-phosphate of AMP, and the other between the 3'-OH of AMP and the 5'-phosphate of GMP; the molecule is called 2'3'-cGAMP.

[0007] Since then, several other patent applications in this area have been published:

[0008] US20140205653 and US 20140341976 of Aduro Biotech disclose cyclic dinucleotide (CDN) compounds that activate and inhibit STING, respectively. Specifically, the CDN of the invention is provided in the form of a composition comprising one or more cyclic purine dinucleotides, which activate or inhibit STING-dependent TBK1 activation and the resulting production of type I interferon.

[0009] WO 2015077354 A1 of the University of Chicago discloses methods and compositions for treating cancer by intratumoral administration of a stimulator of interferon genes (STING) agonist. In some embodiments, compositions and methods for treating cancer in a subject are provided, comprising administering an effective amount of a stimulator of interferon genes (STING) agonist to the subject, wherein the STING agonist is administered intratumorally.

[0010] WO 2015161762 of Fudan University discloses the use of cyclic dinucleotide cGAMP for preparing anti-tumor drugs, wherein the tumor is gastric cancer, lung cancer, colon cancer, liver cancer, prostate cancer or pancreatic cancer. It is shown that cGAMP inhibits the growth of human tumor cell lines in immune-deficient mice.

[0011] WO 2015185565 of GlaxoSmithKline discloses a class of cyclic dinucleotide compounds or pharmaceutically acceptable salts and tautomers thereof, compositions, combinations and drugs containing the compounds, and methods for preparing the same. The invention also relates to the use of the compounds, combinations, compositions and drugs in treating diseases and conditions (e.g., inflammation, allergic and autoimmune diseases, infectious diseases, cancer) in which regulation of STING is beneficial, and as vaccine adjuvants.

[0012] WO 2014179335 to Memorial Sloan Kettering Cancer Center discloses compositions, methods, kits and assays related to the use and / or development of isomers of cGAMP and the structure of the enzyme cGAS.

[0013] There is still a need to discover and develop new cyclic dinucleotide cGAMP analogs for medical treatment. Specifically, there is still a need for cGAMP analogs with better potency, stability and specificity than endogenous cGAMP. No cGAMP analogs have been developed that have excellent safety and efficacy in animal models of human diseases, including cancer and infectious diseases. Summary of the invention

[0014] Formula I includes Formulas Ia-Ii.

[0015] In one aspect, the present invention provides a compound of Formula Ia or a pharmaceutically acceptable salt thereof:

[0016]

[0017] in:

[0018] a and b are independently 0 or 1 and a+b=1, when a is 1, b is 0, and R 5 does not exist; and when a is 0, b is 1, and R 4 does not exist;

[0019] X 1 and X 2 In the five-membered ring, O, S or Se are independently selected;

[0020] From X 1 The alpha carbon starts with L 1 and from X 2 The alpha carbon starts with L 2 are independently -CH2O-P(O)R 6 -O-, -CH2O-P(S)R 6 -O-、-C(Y 1 )(Y 2)OP(O)R 6 -C(Y 3 )(Y 4 )-、-CH2NHSO2NH-、-CH2NHC(O)NH-、-CH2NHC(S)NH-、-CH2NHC(NH)NH-、-CH2NHC(O)CH2-、-CH2NHSO2CH2-、-CH2CH2C(O)NH-、-CH2CH2SO2NH-、-CH2NH(3,4-dioxocyclobutene-1,2-diyl)NH-、

[0021] c is 0, 1, or 2;

[0022] d, e and f are independently 0 or 1;

[0023] Y 1 , Y 2 , Y 3 and Y 4 are independently H or F;

[0024] R 6 is hydroxyl, thiol, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), borano(-BH3 – ) or -NR 7 R 8 ;

[0025] R 7 and R 8 are independently hydrogen, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6Alkylamino, di(C 1-6 Alkyl)amino or azido functionalized C 1-6 Alkyl, cyclo-(C 1-6 alkyl)-, optionally substituted with one or more halogen, mercapto, hydroxyl, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 alkyl)-, cyclo-(C 1-6 Oxyalkyl)-, or optionally one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 Oxaalkyl)-;

[0026] R 1 and R 2 is independently an aromatic or heteroaromatic ring having the following general formula, including tautomeric forms thereof:

[0027]

[0028] g and h are independently 0 or 1;

[0029] W 1 and W 2 are independently hydrogen, halogen, hydroxyl, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 ;

[0030] Z1 , Z 2 , Z 3 , Z 4 , Z 5 and Z 6 are independently CH or N;

[0031] If present, Z 7 , Z 8 , Z 9 , Z 10 and Z 11 are independently CH or N, then W 1 is CH or N; and

[0032] R 3 , R 4 and R 5 are independently hydrogen, halogen, hydroxy, amino, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 .

[0033] In one embodiment, the present invention provides a compound of Formula Ib or a pharmaceutically acceptable salt thereof:

[0034]

[0035] in:

[0036] X 1 and X 2 are independently O, S or Se;

[0037] Z 12 , Z 13 , Z 14 , Z 15 , Z 16 and Z17 are independently CH or N;

[0038] From X 1 The alpha carbon starts with L 1 and from X 2 The alpha carbon starts with L 2 are independently -CH2O-P(O)R 6 -O-, -CH2O-P(S)R 6 -O-、-C(Y 1 )(Y 2 )OP(O)R 6 -C(Y 3 )(Y 4 )-、-CH2NHSO2NH-、-CH2NHC(O)NH-、-CH2NHC(S)NH-、-CH2NHC(NH)NH-、-CH2NHC(O)CH2-、-CH2NHSO2CH2-、-CH2CH2C(O)NH-、-CH2CH2SO2NH-、-CH2NH(3,4-dioxocyclobutene-1,2-diyl)NH-、

[0039]

[0040] c is 0, 1, or 2;

[0041] d, e and f are independently 0 or 1;

[0042] Y 1 , Y 2 , Y 3 and Y 4 are independently H or F;

[0043] R 6 is hydroxyl, thiol, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), borane radical (-BH3 – ) or -NR 7 R 8 ;

[0044] R 7 and R 8 are independently hydrogen, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, cyclo-(C 1-6 alkyl)-, optionally substituted with one or more halogen, mercapto, hydroxyl, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 alkyl)-, cyclo-(C 1-6 Oxyalkyl)-, or optionally one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 Oxaalkyl)-; and

[0045] R 3 and R 4 are independently hydrogen, halogen, hydroxy, amino, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 .

[0046] In another embodiment, the present invention provides a compound of Formula Ic or a pharmaceutically acceptable salt thereof:

[0047]

[0048] in:

[0049] Z 12 , Z 13 , Z 14 , Z 15 , Z 16 and Z 17 are independently CH or N;

[0050] R 3 and R 4 are independently hydrogen, halogen, hydroxy, amino, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 ;

[0051] R 7 and R 8 are independently hydrogen, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, cyclo-(C 1-6alkyl)-, optionally substituted with one or more halogen, mercapto, hydroxyl, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 alkyl)-, cyclo-(C 1-6 Oxyalkyl)-, or optionally one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 Oxaalkyl)-; and

[0052] R 9 and R 10 are independently hydroxyl, thiol, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), borane radical (-BH3 – ) or -NR 7 R 8 .

[0053] In another embodiment, the oxygen atoms in one or both tetrahydrofuran rings of Formula Ic are replaced by sulfur or selenium atoms.

[0054] In another embodiment, the present invention provides a compound of Formula Id or a pharmaceutically acceptable salt thereof:

[0055]

[0056] in:

[0057] X 1 and X2 are independently O, S or Se;

[0058] From X 1 The alpha carbon starts with L 1 and from X 2 The alpha carbon starts with L 2 are independently -CH2O-P(O)R 6 -O-, -CH2O-P(S)R 6 -O-、-C(Y 1 )(Y 2 )OP(O)R 6 -C(Y 3 )(Y 4 )-、-CH2NHSO2NH-、-CH2NHC(O)NH-、-CH2NHC(S)NH-、-CH2NHC(NH)NH-、-CH2NHC(O)CH2-、-CH2NHSO2CH2-、-CH2CH2C(O)NH-、-CH2CH2SO2NH-、-CH2NH(3,4-dioxocyclobutene-1,2-diyl)NH-、

[0059] c is 0, 1, or 2;

[0060] d, e and f are independently 0 or 1;

[0061] Y 1 , Y 2 , Y 3 and Y 4 are independently H or F;

[0062] R 6 is hydroxyl, thiol, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), borane radical (-BH3– ) or -NR 7 R 8 ;

[0063] R 7 and R 8 are independently hydrogen, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, cyclo-(C 1-6 alkyl)-, optionally substituted with one or more halogen, mercapto, hydroxyl, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 alkyl)-, cyclo-(C 1-6 Oxyalkyl)-, or optionally one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 Oxaalkyl)-;

[0064] R 3 and R 4 are independently hydrogen, halogen, hydroxy, amino, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR7 R 8 ;

[0065] R 11 and R 12 Independently selected from:

[0066] Where R 11 and R 12 At least one of

[0067] Z 12 , Z 13 , Z 14 , Z 15 , Z 16 , Z 17 , Z 18 , Z 19 , Z 20 , Z 21 , Z 22 , Z 23 , Z 24 , Z 25 , Z 26 , Z 27 , Z 28 , Z 29 , Z 30 , Z 31 , Z 32 , Z 33 , Z 34 , Z 35 , Z 36 and Z 37 are each independently CH or N; and

[0068] W 3 , W 4 , W 5 , W 6 , W 7 , W 8 and W 9 are independently hydrogen, halogen, hydroxyl, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 .

[0069] In another embodiment, the present invention provides a compound of Formula Ie or a pharmaceutically acceptable salt thereof:

[0070]

[0071] in:

[0072] X 3 , X 4 , X 5 and X 6 are independently O, NH, CH2, CHF or CF2;

[0073] R 13 and R 14 Independently selected from:

[0074]

[0075] Z 12 , Z 13 , Z 14 , Z 15 , Z 16 , Z 17 , Z 18 , Z 19 , Z 20 , Z 21 , Z 22 , Z 23 , Z 24 , Z 25 , Z 26 , Z 27 and Z 28 are each independently CH or N; and

[0076] W 3 , W 4 , W 5 , W 6 and W 7 are independently hydrogen, halogen, hydroxyl, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 ;

[0077] R 7 and R 8 are independently hydrogen, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, cyclo-(C 1-6 alkyl)-, optionally substituted with one or more halogen, mercapto, hydroxyl, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 alkyl)-, cyclo-(C 1-6 Oxyalkyl)-, or optionally one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 Oxaalkyl)-;

[0078] R 3 and R 4 are independently hydrogen, halogen, hydroxy, amino, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 ;and

[0079] R 15 and R 16 are independently hydroxy, mercapto, methoxy, ethoxy, amino, N-methylamino, N,N-dimethylamino, N-ethylamino, N,N-diethylamino, N-morpholinyl or borane (-BH3 – ).

[0080] In another embodiment, the oxygen atoms in one or both tetrahydrofuran rings of Formula le are replaced by sulfur or selenium atoms.

[0081] In another embodiment, the compound is:

[0082]

[0083] In another embodiment, the compound is:

[0084]

[0085] In another embodiment, the compound is:

[0086]

[0087] In another embodiment, the compound is:

[0088]

[0089] In another embodiment, the compound is:

[0090]

[0091] In another embodiment, the compound is:

[0092]

[0093] In another embodiment, the compound is:

[0094]

[0095] In another embodiment, the compound is:

[0096]

[0097] In another embodiment, the compound is:

[0098]

[0099] In another embodiment, the compound is:

[0100]

[0101] In another embodiment, the compound is:

[0102]

[0103] In another embodiment, the compound is:

[0104]

[0105] In another embodiment, the compound is:

[0106]

[0107] In another embodiment, the compound is:

[0108]

[0109] In another embodiment, the compound is:

[0110]

[0111] In another embodiment, the compound is:

[0112]

[0113] In another embodiment, the compound is:

[0114]

[0115] In another embodiment, the compound is:

[0116]

[0117] In another embodiment, the compound is:

[0118]

[0119] In another embodiment, the compound is:

[0120]

[0121] In another embodiment, the compound is:

[0122]

[0123] In one embodiment, the present invention provides a compound of formula If or a pharmaceutically acceptable salt thereof:

[0124]

[0125] in:

[0126] X 1 and X 2 are independently O, S or Se;

[0127] Z 12 , Z 13 , Z 14 , Z 15 , Z 16 and Z 17 are independently CH or N;

[0128] From X 1 The alpha carbon starts with L 1 and from X 2 The alpha carbon starts with L 2 are independently -CH2O-P(O)R 6 -O-, -CH2O-P(S)R 6 -O-、-C(Y 1 )(Y 2 )OP(O)R 6 -C(Y 3 )(Y 4 )-、-CH2NHSO2NH-、-CH2NHC(O)NH-、-CH2NHC(S)NH-、-CH2NHC(NH)NH-、-CH2NHC(O)CH2-、-CH2NHSO2CH2-、-CH2CH2C(O)NH-、-CH2CH2SO2NH-、-CH2NH(3,4-dioxocyclobutene-1,2-diyl)NH-、

[0129] c is 0, 1, or 2;

[0130] d, e and f are independently 0 or 1;

[0131] Y 1 , Y 2 , Y3 and Y 4 are independently H or F;

[0132] R 6 is hydroxyl, thiol, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), borane radical (-BH3 – ) or -NR 7 R 8 ;

[0133] R 7 and R 8 are independently hydrogen, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, cyclo-(C 1-6 alkyl)-, optionally substituted with one or more halogen, mercapto, hydroxyl, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 alkyl)-, cyclo-(C 1-6 Oxyalkyl)-, or optionally one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6Oxaalkyl)-; and

[0134] R 3 and R 5 are independently hydrogen, halogen, hydroxy, amino, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 .

[0135] In one embodiment, the present invention provides a compound of formula Ig or a pharmaceutically acceptable salt thereof:

[0136]

[0137] in:

[0138] Z 12 , Z 13 , Z 14 , Z 15 , Z 16 and Z 17 are independently CH or N;

[0139] R 3 and R 5 are independently hydrogen, halogen, hydroxy, amino, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 ;

[0140] R 7 and R 8 are independently hydrogen, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, cyclo-(C 1-6 alkyl)-, optionally substituted with one or more halogen, mercapto, hydroxyl, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 alkyl)-, cyclo-(C 1-6 Oxyalkyl)-, or optionally one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 Oxaalkyl)-; and

[0141] R 9 and R 10 are independently hydroxyl, thiol, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), borane radical (-BH3 – ) or -NR 7 R 8 .

[0142] In another embodiment, the oxygen atoms in one or both tetrahydrofuran rings of Formula Ig are replaced with sulfur or selenium atoms.

[0143] In another embodiment, the present invention provides a compound of Formula Ih or a pharmaceutically acceptable salt thereof:

[0144]

[0145] in:

[0146] X 1 and X 2 are independently O, S or Se;

[0147] From X 1 The alpha carbon starts with L 1 and from X 2 The alpha carbon starts with L 2 are independently -CH2O-P(O)R 6 -O-, -CH2O-P(S)R 6 -O-、-C(Y 1 )(Y 2 )OP(O)R 6 -C(Y 3 )(Y 4 )-、-CH2NHSO2NH-、-CH2NHC(O)NH-、-CH2NHC(S)NH-、-CH2NHC(NH)NH-、-CH2NHC(O)CH2-、-CH2NHSO2CH2-、-CH2CH2C(O)NH-、-CH2CH2SO2NH-、-CH2NH(3,4-dioxocyclobutene-1,2-diyl)NH-、

[0148] c is 0, 1, or 2;

[0149] d, e and f are independently 0 or 1;

[0150] Y 1 , Y 2 , Y 3 and Y 4 are independently H or F;

[0151] R 6 is hydroxyl, thiol, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), borane radical (-BH3 – ) or -NR 7 R 8 ;

[0152] R 7 and R 8 are independently hydrogen, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, cyclo-(C 1-6 alkyl)-, optionally substituted with one or more halogen, mercapto, hydroxyl, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 alkyl)-, cyclo-(C 1-6 Oxyalkyl)-, or optionally one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 Oxaalkyl)-;

[0153] R3 and R 5 are independently hydrogen, halogen, hydroxy, amino, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 ;

[0154] R 11 and R 12 Independently selected from:

[0155] Where R 11 and R 12 At least one of

[0156] Z 12 , Z 13 , Z 14 , Z 15 , Z 16 , Z 17 , Z 18 , Z 19 , Z 20 , Z 21 , Z 22 , Z 23 , Z 24 , Z 25 , Z 26 , Z 27 , Z 28 , Z 29 , Z 30 , Z 31 , Z 32 , Z 33 , Z 34 , Z 35 , Z 36and Z 37 are each independently CH or N; and

[0157] W 3 , W 4 , W 5 , W 6 , W 7 , W 8 and W 9 are independently hydrogen, halogen, hydroxyl, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 .

[0158] In another embodiment, the present invention provides a compound of Formula Ii or a pharmaceutically acceptable salt thereof:

[0159]

[0160] in:

[0161] X 3 , X 5 , X 6 and X 7 are independently O, NH, CH2, CHF or CF2;

[0162] R 13 and R 14 Independently selected from:

[0163]

[0164] Z 12 , Z 13 , Z 14 , Z 15 , Z 16 , Z 17, Z 18 , Z 19 , Z 20 , Z 21 , Z 22 , Z 23 , Z 24 , Z 25 , Z 26 , Z 27 and Z 28 are each independently CH or N; and

[0165] W 3 , W 4 , W 5 , W 6 and W 7 are independently hydrogen, halogen, hydroxyl, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 ;

[0166] R 7 and R 8 are independently hydrogen, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, cyclo-(C 1-6 alkyl)-, optionally substituted with one or more halogen, mercapto, hydroxyl, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 alkyl)-, cyclo-(C 1-6 Oxyalkyl)-, or optionally one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl)amino functionalized ring-(C 1-6 Oxaalkyl)-;

[0167] R 3 and R 5 are independently hydrogen, halogen, hydroxy, amino, C 1-6 Alkyl, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Alkoxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkyl, C 1-6 Alkoxy, optionally substituted with one or more halogen, mercapto, hydroxy, carbonyl, carboxyl, carbonyloxy, C 1-6 Hydroxyalkoxy, amino, C 1-6 Alkylamino, di(C 1-6 C 1-6 Alkoxy, C 3-5 Alkenyl-O-, C 3-5 Alkynyl-O-, oligo(ethylene glycol), poly(ethylene glycol), azido or -NR 7 R 8 ;and

[0168] R 15 and R 16 are independently hydroxy, mercapto, methoxy, ethoxy, amino, N-methylamino, N,N-dimethylamino, N-ethylamino, N,N-diethylamino, N-morpholinyl or borane (-BH3 – ).

[0169] In another embodiment, the oxygen atoms in one or both tetrahydrofuran rings of Formula Ii are replaced with sulfur or selenium atoms.

[0170] In another embodiment, the compound is:

[0171]

[0172] In another embodiment, the compound is:

[0173]

[0174] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

[0175] In another aspect, the present invention provides a method of treating a disease or condition in which modulation of STING is beneficial, comprising: administering a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof.

[0176] In another aspect, the present invention provides a compound of Formula I or a pharmaceutically acceptable salt thereof for use in treating a disease or condition in which modulation of STING would be beneficial.

[0177] In another aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof for use in therapy.

[0178] In another aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof for use in treating cancer.

[0179] In another aspect, the present invention provides a compound of Formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, such as a nanoparticle or a delivery vehicle, which enhances the cellular uptake, stability and efficacy of the compound of Formula I for treating cancer.

[0180] In another aspect, the present invention provides a method for treating cancer, comprising: administering a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof.

[0181] In another aspect, the present invention provides use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer.

[0182] In another aspect, the present invention provides a pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent.

[0183] In another aspect, the present invention provides a pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, at least one other therapeutic agent and one or more pharmaceutically acceptable excipients.

[0184] In another aspect, the present invention provides a pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent, for use in treatment.

[0185] In another aspect, the present invention provides a pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent for use in treating a disease or condition in which modulation of STING is beneficial.

[0186] In another aspect, the present invention provides a pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent, for use in treating cancer.

[0187] In another aspect, the present invention provides a method of treating a disease or condition in which modulation of STING is beneficial, comprising: administering to a patient in need thereof a therapeutically effective amount of a combination comprising a compound of Formula I or a pharmaceutically acceptable salt and at least one other therapeutic agent.

[0188] In another aspect, the present invention provides a method for treating cancer, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition, wherein the pharmaceutical composition comprises: a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent.

[0189] In another aspect, the present invention provides a pharmaceutical composition comprising: a compound of formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent for treating cancer. The therapeutic agent includes but is not limited to immune checkpoint inhibitors, such as humanized antibodies against PD1, PD-L1, CTLA4, and other molecules that block effective anti-tumor immune responses.

[0190] In another aspect, the present invention provides a method for treating cancer, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition, the pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent. The therapeutic agent includes but is not limited to immune checkpoint inhibitors, such as humanized antibodies against PD1, PD-L1, CTLA4, and other molecules that block effective anti-tumor immune responses.

[0191] In another aspect, the present invention provides a pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent for treating cancer. The therapeutic agent includes radiation, such as high-dose radiation, which directly kills tumor cells, enhances the presentation of tumor antigens and activates the STING pathway.

[0192] In another aspect, the present invention provides a method for treating cancer, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent. The therapeutic agent includes radiation, such as high-dose radiation, which directly kills tumor cells, enhances the presentation of tumor antigens and activates the STING pathway.

[0193] In another aspect, the present invention provides a pharmaceutical composition comprising: a compound of formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent for treating cancer. The therapeutic agent includes another chemotherapeutic agent that selectively kills tumor cells and enhances the presentation of tumor antigens.

[0194] In another aspect, the present invention provides a method for treating cancer, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition, the pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent. The therapeutic agent comprises another chemotherapeutic agent that selectively kills tumor cells and enhances the presentation of tumor antigens.

[0195] In another aspect, the present invention provides a pharmaceutical composition comprising: a compound of formula I or a pharmaceutically acceptable salt thereof, a pharmaceutical preparation comprising nanoparticles, and at least one other therapeutic agent for treating cancer. The therapeutic agent comprises radiation and / or another chemotherapeutic agent.

[0196] In another aspect, the present invention provides a method for treating cancer, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, a pharmaceutical preparation comprising nanoparticles, and at least one other therapeutic agent for treating cancer. The therapeutic agent comprises radiation and / or another chemotherapeutic agent.

[0197] On the other hand, the present invention provides a method for treating cancer, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition, the pharmaceutical composition comprising: a compound of Formula I or a pharmaceutically acceptable salt thereof, a pharmaceutical preparation comprising nanoparticles, and at least one other therapeutic agent, for treating cancer. The compound of Formula I can be injected directly into the tumor or systemically, including injection into muscle (intramuscular), skin (subcutaneous and intradermal), peritoneum (intraperitoneal), lymph node (intralymphatic) or vein (intravenous).

[0198] In another aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof for use as a vaccine adjuvant.

[0199] In another aspect, the present invention provides a compound of Formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, such as a nanoparticle or a delivery vehicle that enhances the cellular uptake, stability and efficacy of the compound of Formula I, which is used as a vaccine adjuvant.

[0200] In one embodiment, the pharmaceutical composition is a vaccine.

[0201] In another embodiment, the present invention provides a method of inducing or promoting an immune response, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof as an adjuvant and a tumor antigen.

[0202] In another embodiment, the present invention provides a method of inducing or promoting an immune response, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formula I, or a pharmaceutical composition thereof as an adjuvant, a tumor antigen, or a pharmaceutical composition thereof, such as a nanoparticle or delivery vehicle that enhances the cellular uptake of the adjuvant and the tumor antigen.

[0203] In another aspect, the present invention provides a pharmaceutical composition comprising: a compound of formula I or a pharmaceutically acceptable salt thereof as an adjuvant and an immunogen of a target pathogen.

[0204] In another aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof for use as a vaccine adjuvant.

[0205] In another embodiment, the present invention provides a method of inducing or promoting an immune response, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof as an adjuvant and an immunogen of a target pathogen.

[0206] In another aspect, the present invention provides a vaccine adjuvant comprising: a compound of formula I or a pharmaceutically acceptable salt thereof.

[0207] In another aspect, the present invention provides an immunogenic composition comprising: an antigen or an antigen composition and a compound of formula I or a pharmaceutically acceptable salt thereof.

[0208] In another aspect, the present invention provides an immunogenic composition comprising: an antigen or an antigen composition and a compound of formula I or a pharmaceutically acceptable salt thereof, for use in treating or preventing diseases, including cancer and infectious diseases.

[0209] In another aspect, the present invention provides use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of an immunogenic composition comprising an antigen or antigenic composition for treating or preventing a disease (including cancer and infectious diseases).

[0210] In another aspect, the present invention provides a method for treating or preventing a disease, comprising: administering an immunogenic composition comprising an antigen or antigenic composition and a compound of formula I or a pharmaceutically acceptable salt thereof to a patient suffering from or susceptible to the disease.

[0211] In another aspect, the present invention provides a vaccine composition comprising: an antigen or an antigen composition and a compound of formula I or a pharmaceutically acceptable salt thereof, for use in treating or preventing diseases, including cancer and infectious diseases.

[0212] In another aspect, the present invention provides use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a vaccine composition for treating or preventing a disease (including cancer and infectious diseases), wherein the vaccine composition comprises an antigen or an antigen composition.

[0213] In another aspect, the present invention provides a method for treating or preventing a disease, comprising administering a vaccine composition comprising an antigen or antigen composition and a compound of formula I or a pharmaceutically acceptable salt thereof to a patient suffering from or susceptible to the disease.

[0214] In another aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof for use in treating immune disorders, including autoimmune diseases and autoinflammatory diseases.

[0215] In another aspect, the present invention provides a compound of Formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, such as a nanoparticle or a delivery vehicle that enhances the cellular uptake, stability and efficacy of the compound of Formula I, for use in treating immune disorders, including autoimmune diseases and autoinflammatory diseases.

[0216] In another aspect, the present invention provides a method for treating an immune disorder, comprising: administering a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof.

[0217] In another aspect, the present invention provides the use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating immune disorders (including autoimmune diseases and autoinflammatory diseases).

[0218] It should be understood that all combinations of the above aspects / embodiments, and other aspects / embodiments disclosed elsewhere herein, are contemplated and are further embodiments of the invention. DETAILED DESCRIPTION

[0219] The present invention provides new cGAMP analogs, pharmaceutical compositions thereof and uses thereof in treatment. 2'3'-cGAMP is an endogenous second messenger produced by mammalian cells. It is a high-affinity ligand of STING, inducing conformational changes therein, and is an effective inducer of type I interferon. cGAS and the cGAS-cGAMP pathway are very important for triggering inflammatory responses to self and exogenous DNA. Therefore, cGAS is important for immune defense against microbial pathogens that contain DNA and require DNA in their life cycle. These pathogens include DNA viruses, retroviruses (including HIV), bacteria (including mycobacterium tuberculosis), fungi and parasites. cGAS can also detect tumor DNA and is important for the body's intrinsic immunity against malignant cells. Activation of the cGAS-cGAMP-STING pathway is important for cancer immunotherapy.

[0220] As an effective inducer of type I interferon, cGAMP (and therefore cGAMP analogs of the present invention) provides a reasonable immune adjuvant. Therefore, the compound of Formula I or a pharmaceutically acceptable salt thereof can be used as a vaccine adjuvant, particularly for use with mucosal vaccines, and can be formulated and delivered together with an immunogen, as cyclic-di-GMP and c-di-AMP, which have been used as vaccine adjuvants (see, for example, Pedersen, et al. PLoS ONE, Nov 2011, 6, 11, e26973; Ebensen et al., Vaccine 29, 2011, 5210-5220; Chen et al., Vaccine 28, 2010, 3080-3085). In fact, such adjuvants are often more effective because cGAMP (and cGAMP analogs of the present invention) is more effective than c-di-GMP in inducing interferon.

[0221] In one aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof for the treatment of cancer. In one embodiment, the pharmaceutical composition is a compound of Formula I. In another embodiment, the pharmaceutical composition is a compound of Formula I in the form of a pharmaceutical preparation comprising nanoparticles or another delivery vehicle. In another embodiment, the pharmaceutical composition is a combination of a compound of Formula I and at least one other therapeutic agent, including but not limited to immune checkpoint inhibitors, such as antibodies against PD-1, PD-L1 or CTLA-4. Therapeutic agents used in combination with compounds of Formula I also include radiation to tumors or chemotherapeutic agents targeting tumor cells.

[0222] On the other hand, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof as an adjuvant and an immunogen of a target pathogen. In one embodiment, the pharmaceutical composition is a vaccine. In another embodiment, the present invention provides a method of inducing or promoting an immune response, comprising: administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof as an adjuvant and an immunogen of a target pathogen.

[0223] As used in this article:

[0224] The terms "halo" and "halogen", alone or in combination with other groups, refer to fluoro-, chloro-, bromo- and iodo- groups.

[0225] The term "C 1-6 "Alkyl", alone or in combination with other groups, refers to a monovalent straight or branched chain alkyl group containing 1 to 6 carbon atoms. Exemplary C 1-6 Alkyl includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl and tert-butyl. More preferred is C 1-4 alkyl.

[0226] The term "C 1-6 "Alkoxy", alone or in combination with other groups, refers to R'-O-, where R' is C 1-6 alkyl.

[0227] The term "halogenated C 1-6 "Alkyl", alone or in combination with other groups, refers to a C 1-6 Alkyl groups, such as CF3 and CH2CF3.

[0228] Unless otherwise indicated, the term "compound of the present invention" or "compound of formula I" includes all solvates, complexes, polymorphs, radiolabeled derivatives, tautomers, stereoisomers and optical isomers of the compound of formula I and its salts.

[0229] The term "effective amount" refers to the amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought, for example, by a researcher or clinician.

[0230] The term "therapeutically effective amount" refers to any amount that causes improved treatment, healing, prevention or amelioration of a disease, condition or side effect, or a reduction in the rate of progression of a disease or condition, compared to a corresponding subject not receiving such amount. The term also includes within its scope amounts that are effective to enhance normal physiological function.

[0231] The term "prophylaxis" includes prevention and refers to measures or procedures that prevent rather than cure or treat a disease. Prevention refers to reducing the risk of acquiring or developing a disease resulting in at least one clinical symptom of the disease rather than identifying subjects who may have been exposed to the causative agent or who are susceptible to the disease before the onset of the disease.

[0232] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0233] The term "pharmaceutically acceptable excipient" includes all diluents, carriers, binders, glidants and other components of pharmaceutical formulations with which the compounds of the present invention are administered.

[0234] The compounds of the present invention may exist in solid or liquid form. In solid form, the compounds of the present invention may exist in a continuum of solid states ranging from completely amorphous to completely crystalline.

[0235] The term "amorphous" refers to a state in which the material lacks long-range order at the molecular level and, depending on the temperature, can exhibit the physical properties of a solid or a liquid. Typically, such materials do not provide a distinctive X-ray diffraction pattern and, while exhibiting the properties of a solid, are more formally described as liquids. Upon heating, a change from solid to liquid properties occurs, characterized by a change of state, usually second order ("glass transition").

[0236] The term "crystalline" refers to a solid phase in which the material has a regularly ordered internal structure at the molecular level and gives a distinctive X-ray diffraction pattern with defined peaks. Such a material will also exhibit the properties of a liquid when heated sufficiently, but the change from solid to liquid is characterized by a phase transition, usually first order ("melting point").

[0237] The compounds of the present invention may have the ability to crystallize in more than one form, a feature known as polymorphism, and it is understood that such multiple forms ("polymorphs") are within the scope of the present invention. Polymorphism may generally occur as a response to changes in temperature or pressure, or both, and may also result from changes in the crystallization process. Polymorphs may be distinguished by various physical characteristics known in the art, such as x-ray diffraction patterns, solubility, and melting points.

[0238] The compound of formula I can exist in solvated and non-solvated forms. As used herein, the term "solvate" refers to a complex of variable stoichiometry formed by a solute (in the present invention, a compound or salt of formula I) and a solvent. These solvents for the purposes of the present invention may not interfere with the biological activity of the solute. It will be appreciated by those skilled in the art that pharmaceutically acceptable solvates can be formed for crystalline compounds, wherein solvent molecules are incorporated into the crystal lattice during the crystallization process. The incorporated solvent molecules can be water molecules or non-water molecules, such as ethanol, isopropanol, DMSO, acetic acid, ethanolamine and ethyl acetate molecules. The crystal lattice incorporating water molecules is generally referred to as a "hydrate". Hydrates include stoichiometric hydrates and compositions containing variable amounts of water. The present invention includes all of these solvates.

[0239] It is also noted that some compounds can form tautomers. "Tautomers" refer to compounds of interchangeable forms of specific compound structures, as well as compounds with hydrogen atom and electronic displacement changes. Therefore, the two structures can be in balance by the movement of re electrons and atoms (usually H). For example, enols and ketones are tautomers because they are rapidly converted to each other by treating with acids or bases. It should be understood that all tautomers of the compounds of the present invention and mixtures of tautomers are included within the scope of the compounds of the present invention.

[0240] The compound of formula I can be in the form of a salt. Generally, the salt of the present invention is a pharmaceutically acceptable salt. The salts covered in the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of the present invention. For a review of suitable salts, see, for example, Berge et al, J. Pharm. Sci. 1977, 66, 1-19. Suitable pharmaceutically acceptable salts may include acid addition salts. Pharmaceutically acceptable acid addition salts may be formed by reacting a compound of formula I with a suitable inorganic acid or organic acid (such as hydrobromic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, p-toluenesulfonic acid, benzenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, naphthalenesulfonic acid such as 2-naphthalenesulfonic acid) in a suitable solvent such as an organic solvent to give a salt, which is often separated, for example, by crystallization and filtration. The pharmaceutically acceptable acid addition salt of the compound of formula I may be, for example, hydrobromide, hydrochloride, sulfate, nitrate, phosphate, p-toluenesulfonate, benzenesulfonate, methanesulfonate, ethanesulfonate or naphthalenesulfonate (e.g., 2-naphthalenesulfonate). Other non-pharmaceutically acceptable salts (eg, trifluoroacetate salts) may be useful, for example, in the isolation of the compounds of the invention and are included within the scope of the invention.

[0241] The present invention includes within its scope all possible stoichiometric and non-stoichiometric forms of the compounds of formula I.

[0242] Although the compounds of the present invention can be administered as raw chemicals for treatment, it is also possible for the compounds of the present invention to be present in a pharmaceutical composition as an active ingredient. Such compositions can be prepared in a manner well known in the pharmaceutical field and contain at least one active compound. Therefore, the present invention also provides a pharmaceutical composition comprising a compound of the present invention and one or more pharmaceutically acceptable excipients. The excipient must be acceptable in the sense that it is compatible with the other ingredients of the composition and harmless to its recipient. According to another aspect of the present invention, a method for preparing a pharmaceutical composition is also provided, the pharmaceutical composition comprising the substance or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. The pharmaceutical composition can be used to treat and / or prevent any condition described herein.

[0243] Typically, the compounds of the present invention are administered in a pharmaceutically effective amount. The amount of the compound actually administered is usually determined by a doctor according to relevant circumstances, including the condition to be treated, the selected route of administration, the actual compound administered, the age, body weight and reaction of individual patients, the severity of the patient's symptoms, etc. The pharmaceutical composition can exist in a unit dosage form containing a predetermined amount of active ingredient per unit dose. The term "unit dosage form" refers to a physically discrete unit suitable as a unit dose for human subjects and other mammals, each unit containing a predetermined amount of active substance and a suitable pharmaceutical excipient, vehicle or carrier calculated to produce the desired therapeutic effect. Typical unit dosage forms include pre-filled, pre-measured ampoules or syringes of liquid compositions, or pills, tablets, capsules, etc. in the case of solid compositions.

[0244] Preferred unit dose compositions are those containing a daily dose or subdose of the active ingredient or an appropriate fraction thereof. Thus, such unit doses may be administered once or more than once per day. Such pharmaceutical compositions may be prepared by any method known in the pharmaceutical art.

[0245] The pharmaceutical composition may be suitable for administration by any appropriate route, such as oral (including buccal or sublingual), rectal, inhalation, intranasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal). Such compositions may be prepared by any method known in the pharmaceutical art, such as by combining the active ingredient with (one or more) carriers or (one or more) excipients.

[0246] Pharmaceutical compositions suitable for oral administration may be presented as discrete units such as capsules or tablets; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or whips; or oil-in-water liquid emulsions or water-in-oil liquid emulsions.

[0247] For example, for oral administration in tablet or capsule form, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert excipient such as ethanol, glycerol, water, etc. Powders are prepared by reducing the compound to a suitable fine size and mixing with a similarly prepared pharmaceutical excipient such as an edible carbohydrate (e.g., starch or mannitol). Flavoring agents, preservatives, dispersants, and coloring agents may also be present.

[0248] Capsules are prepared by preparing a powder mixture as described above and filling the formed gelatin sheath. Excipients, including glidants and lubricants, such as colloidal silicon dioxide, talc, magnesium stearate, calcium stearate or solid polyethylene glycol, may be added to the powder mixture prior to the filling operation. Disintegrants or solubilizers such as agar, calcium carbonate or sodium carbonate may also be added to improve the availability of the drug when the capsule is ingested.

[0249] Moreover, excipients (including suitable binders, glidants, lubricants, sweeteners, flavors, disintegrants and coloring agents) may also be incorporated into the mixture when needed or necessary. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as gum arabic, tragacanth or sodium alginate, carboxymethyl cellulose, polyethylene glycol, waxes, etc. The lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, etc. Disintegrants include but are not limited to starch, methylcellulose, agar, bentonite, xanthan gum, etc. Tablets are prepared, for example, by preparing a powder mixture, granulating or slugging, adding lubricants and disintegrants and pressing into tablets. The powder mixture is prepared by mixing the appropriately pulverized compound with a diluent or alkali as described above, and optionally with a binder (such as carboxymethylcellulose, alginate, gelatin or polyvinyl pyrrolidone), a solvent retardant (such as paraffin), a reabsorption promoter (such as a quaternary salt) and / or an absorbent (such as bentonite, kaolin or dicalcium phosphate). The powder mixture can be granulated by wetting with a binder such as syrup, starch paste, Acadia mucilage or a solution of cellulose or polymer material and applying force to sieve. As an alternative to granulation, the powder mixture can be run through a tablet press, and the result is that the incompletely formed lumps are broken into granules. The granules can be lubricated by adding stearic acid, stearate, talc or mineral oil to prevent adhesion to the tablet forming die. The lubricated mixture is then pressed into tablets. The compounds of the present invention can also be combined with a free-flowing inert carrier and directly pressed into tablets without granulation or punching steps. A transparent or opaque protective coating can be provided, which consists of a coating of shellac sealing, a coating of sugar or polymer material and a polishing coating of wax. Dyestuffs can be added to these coatings to distinguish different unit dosages.

[0250] Oral fluids such as solutions, suspensions, syrups and elixirs can be prepared in dosage unit form so that a given amount contains a predetermined amount of compound. Syrups can be prepared by dissolving the compound in an aqueous solution of appropriate flavoring, and elixirs are prepared by using non-toxic alcohol carriers. Suspensions can be prepared by dispersing the compound in a non-toxic carrier. Solubilizers and emulsifiers such as ethoxylated isostearyl alcohol and polyoxyethylene sorbitol ethers, preservatives, flavor additives such as peppermint oil or natural sweeteners or saccharin or other artificial sweeteners, etc. can also be added.

[0251] Where appropriate, dosage unit compositions for oral administration can be microencapsulated. The compositions can also be prepared to prolong or sustain the release, for example, by coating or embedding particulate material in polymers, wax, etc.

[0252] The compounds of the invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholine. Pharmaceutical compositions suitable for transdermal administration can be presented as discrete patches that are intended to remain in close contact with the epidermis of the recipient for a long period of time.

[0253] Pharmaceutical compositions adapted for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils.

[0254] For treatment of the eye or other external tissues (e.g., oral cavity and skin), the composition is preferably applied as a topical ointment or cream. In the preparation of an ointment, the active ingredient may be used with a paraffin or water-miscible ointment base. Alternatively, the active ingredient may be formulated in an ointment with an oil-in-water cream base or a water-in-oil base.

[0255] Pharmaceutical compositions adapted for topical administration to the eye include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent.

[0256] Pharmaceutical compositions adapted for topical administration in the mouth include lozenges, pastilles and mouthwashes.

[0257] Pharmaceutical compositions adapted for rectal administration may be presented as suppositories or as enemas.

[0258] Dosage forms for nasal or inhaled administration may conveniently be formulated as aerosols, solutions, suspension drops, gels or dry powders.

[0259] Compositions for intranasal administration include aqueous compositions applied to the nose by drops or by a pressurized pump. Suitable compositions contain water as a diluent or carrier for this purpose. Compositions for application to the lungs or nose may contain one or more excipients, such as one or more suspending agents, one or more preservatives, one or more surfactants, one or more tension modifiers, one or more cosolvents, and may include components that control the pH of the composition, such as a buffer system. In addition, the composition may contain other excipients, such as antioxidants, such as sodium metabisulfite; and taste masking agents. The composition may also be applied to other areas of the nose or respiratory tract by atomization. Intranasal compositions may allow the compound of formula I or a pharmaceutically acceptable salt thereof to be delivered to all areas of the nasal cavity (target tissues), and further may allow the compound of formula I or a pharmaceutically acceptable salt thereof to remain in contact with the target tissue for a longer period of time. A suitable dosage regimen for intranasal compositions is that the patient slowly inhales through the nose after the nasal cavity is cleaned. During inhalation, the composition is applied to one nostril, while the other nostril is pressed by hand. The procedure is then repeated for the other nostril. Typically, one or two sprays are applied to each nostril once, twice or three times daily, ideally once daily, by the above method. Of particular interest are intranasal compositions suitable for once-daily administration.

[0260] Suspending agents, if included, are typically present in an amount of 0.1 to 5% (w / w), such as 1.5 to 2.4% (w / w), based on the total weight of the composition. Examples of pharmaceutically acceptable suspending agents include, but are not limited to, Avicef (microcrystalline cellulose and sodium carboxymethylcellulose), sodium carboxymethylcellulose, magnesium aluminum silicate (veegum), tragacanth, bentonite, methylcellulose, xanthan gum, carbomer, and polyethylene glycol.

[0261] Compositions for application to the lungs or noses may contain one or more excipients, and may be protected from microbial or fungal contamination and growth by including one or more preservatives. Examples of pharmaceutically acceptable antimicrobial agents or preservatives include, but are not limited to, quaternary ammonium compounds (e.g., benzalkonium chloride, benzethonium chloride, cetrimonium bromide, cetylpyridinium chloride, laurochloramine, and myristylpyridinium chloride), mercurials (e.g., phenylmercuric nitrate, phenylmercuric acetate, and thimerosal), alcohols (e.g., chlorobutanol, phenylethyl alcohol, and benzyl alcohol), antibacterial esters (e.g., esters of parahydroxybenzoic acid), chelating agents such as disodium ethylenediaminetetraacetic acid (EDTA), and other antibacterial agents such as chlorhexidine, chlorocresol, sorbic acid and its salts (e.g., potassium sorbate), and polymyxins. Examples of pharmaceutically acceptable antifungal agents or preservatives include, but are not limited to, sodium benzoate, sorbic acid, sodium propionate, methylparaben, ethylparaben, propylparaben, and butylparaben. The preservative, if included, may be present in an amount of 0.001 to 1% (w / w), such as 0.015% to 0.5% (w / w), based on the total weight of the composition. The composition (e.g., wherein at least one compound is in suspension) may include one or more surfactants that are used to promote the dissolution of the drug particles in the aqueous phase of the composition. For example, the amount of surfactant used is an amount that does not cause foaming during the mixing process. Examples of pharmaceutically acceptable surfactants include fatty alcohols, esters and ethers, such as polyoxyethylene (20) sorbitan monooleate (polysorbate 80), polyethylene glycol ethers and poloxamers. The surfactant may be present in an amount of about 0.01 to 10% (w / w), such as 0.01 to 0.75% (w / w), such as about 0.5% (w / w), based on the total weight of the composition.

[0262] One or more tonicity regulators may be included to achieve tonicity of body fluids (e.g., nasal fluids), resulting in reduced irritation levels. Examples of pharmaceutically acceptable tonicity regulators include, but are not limited to, sodium chloride, dextrose, xylitol, calcium chloride, glucose, glycerol, and sorbitol. If present, the tonicity regulator may be included in an amount of 0.1 to 10% (w / w), such as 4.5 to 5.5% (w / w), such as about 5.0% (w / w), based on the total weight of the composition.

[0263] The compositions of the present invention may be buffered by the addition of a suitable buffer such as sodium citrate, citric acid, tromethamine, phosphates such as disodium phosphate (eg, dodecahydrate, heptahydrate, dihydrate and anhydrous forms) or sodium phosphate and mixtures thereof.

[0264] If present, the buffer may be included in an amount of 0.1 to 5% (w / w), for example 1 to 3% (w / w), based on the total weight of the composition.

[0265] Examples of taste masking agents include sucralose, sucrose, saccharin or its salts, fructose, dextrose, glycerin, corn syrup, aspartame, acesulfame-K, xylitol, sorbitol, erythritol, ammonium glycyrrhizinate, thaumatin, neotame, mannitol, menthol, eucalyptus oil, camphor, natural flavorings, artificial flavorings, and combinations thereof.

[0266] One or more cosolvents may be included to help dissolve the drug compound and / or other excipients. Examples of pharmaceutically acceptable cosolvents include, but are not limited to, propylene glycol, dipropylene glycol, ethylene glycol, glycerol, ethanol, polyethylene glycol (e.g., PEG300 or PEG400) and methanol. In one embodiment, the cosolvent is propylene glycol.

[0267] If present, the co-solvent may be included in an amount of 0.05 to 30% (w / w), such as 1 to 25% (w / w), for example 1 to 10% (w / w), based on the total weight of the composition.

[0268] Compositions for inhalation administration include aqueous, organic or aqueous / organic mixtures, dry powders or crystalline compositions applied to the respiratory tract by a pressurized pump or inhaler (e.g., a reservoir dry powder inhaler, a unit dose dry powder inhaler, a pre-metered multi-dose dry powder inhaler, a nasal inhaler or a pressurized aerosol inhaler, a nebulizer or an insufflator). Suitable compositions contain water as a diluent or carrier for this purpose, and conventional excipients, such as buffers, tension modifiers, etc., may also be provided. Aqueous compositions may also be applied to other areas of the nose and respiratory tract by atomization. Such compositions may be aqueous solutions or suspensions or aerosols delivered from pressurized packaging (e.g., metered dose inhalers) using suitable liquefied propellants.

[0269] Compositions for topical application to the nose (e.g., for the treatment of rhinitis) or the lungs include pressurized aerosol compositions and aqueous compositions delivered to the nasal cavity by a pressurized pump. Compositions that are non-pressurized and suitable for topical application to the nasal cavity are of particular interest. Suitable compositions contain water as a diluent or carrier for this purpose. Aqueous compositions for application to the lungs or nose can be provided with conventional excipients, such as buffers, tension regulators, etc. Aqueous compositions can also be applied to the nose by atomization.

[0270] Fluid distributor can be used for delivering fluid composition to nasal cavity usually.Fluid composition can be aqueous or non-aqueous, but is aqueous usually.This fluid distributor can have distribution nozzle or distribution hole, when the force applied by the user is applied to the pump mechanism of fluid distributor, distribute the fluid composition of metered dose by this distribution nozzle or distribution hole.This fluid distributor is usually provided with the reservoir of the fluid composition of multiple metered doses, and these doses are distributable after successive pump actuation.Distribution nozzle or distribution hole can be configured to be inserted in the nostril of user, so that fluid composition spray is distributed in nasal cavity.

[0271] Dry powder compositions for topical delivery to the lungs by inhalation can be presented, for example, in capsules and cartridges of, for example, gelatin or in blisters of, for example, laminated aluminum foil for use in inhalers or insufflators. Powder mixture compositions typically contain a compound of Formula I or a pharmaceutically acceptable salt thereof and a suitable powder matrix (carrier / diluent / excipient material) such as a monosaccharide, disaccharide or polysaccharide (e.g., lactose or starch) for inhalation powder mixture. In addition to the drug and carrier, the dry powder composition can also include additional excipients (e.g., ternary materials such as sugar esters, such as cellobiose octaacetate, calcium stearate or magnesium stearate.

[0272] Pharmaceutical compositions suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions, which may contain antioxidants, buffers, bacteriostats and solutes that make the composition isotonic with the blood of the intended recipient; aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickening agents. The composition may be presented in unit dose or multi-dose containers, such as sealed ampoules and vials, and may be stored under freeze-dried (lyophilized) conditions, requiring only the addition of a sterile liquid carrier, such as water for injection, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.

[0273] It should be understood that in addition to the ingredients particularly mentioned above, the compositions may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents.

[0274] The compounds of formula I and their pharmaceutically acceptable salts can also be formulated with other adjuvants to modulate their activity. Such compositions may contain antibodies or antibody fragments or antigenic components, including but not limited to proteins, DNA, live or dead bacteria and / or viruses or virus-like particles; and one or more components with adjuvant activity, including but not limited to aluminum salts, oil and water emulsions, heat shock proteins, lipid A preparations and derivatives, glycolipids, other TLR agonists such as CpG DNA or similar substances, cytokines such as GM-CSF or IL-12 or similar substances.

[0275] The therapeutically effective amount of the agent will depend on many factors, including, for example, the age and weight of the subject, the precise condition requiring treatment and its severity, the nature of the formulation and the route of administration, and will ultimately be at the discretion of the attending physician or veterinarian. In particular, the subject to be treated is a mammal, particularly a human.

[0276] The medicament can be administered in a daily dose. The amount can be given in a single dose per day, or more usually, in several (e.g., two, three, four, five or six) sub-doses per day so that the total daily dose is the same.

[0277] Suitably, the amount of the compound of the invention to be administered according to the present invention will be an amount selected from about 0.01 mg to about 1 g per day (calculated as free or unsalted compound).

[0278] The compound of formula I and its pharmaceutically acceptable salt can be used alone or in combination with other therapeutic agents. The compound of formula I and its pharmaceutically acceptable salt and other pharmaceutically active agents can be applied together or separately. When applied separately, the application can be applied simultaneously or sequentially in any order by any convenient route with a single or combined pharmaceutical composition. The amount of the compound of formula I or its pharmaceutically acceptable salt and other pharmaceutically active agents and the relevant application timing are selected to achieve the desired combined therapeutic effect. The compound of formula I or its pharmaceutically acceptable salt and other therapeutic agents can be used in combination by applying simultaneously in a unit pharmaceutical composition including these two compounds. Alternatively, the combination can be applied separately in different pharmaceutical compositions, and each pharmaceutical composition includes a compound in a sequential manner, wherein, for example, the compound of the present invention is first applied, and then the second compound is applied, and vice versa. This sequential application can be close in time (for example, at the same time) or at a relatively long time interval. In addition, it does not matter whether the compound is applied in the same dosage form, for example, one compound can be applied topically, and another compound can be applied orally. Suitably, both compounds are applied orally.

[0279] The combination can be presented as a combination kit. The term "combination kit" or "kit" as used herein refers to one or more pharmaceutical compositions for administering the combination of the present invention. When two compounds are administered simultaneously, the combination kit may contain the two compounds in a single pharmaceutical composition (such as a tablet) or in different pharmaceutical compositions. When the compounds are not administered simultaneously, the combination kit will contain each compound in different pharmaceutical compositions in a single package or in different pharmaceutical compositions in different packages. The combination kit may also be provided with instructions, such as dosage and administration instructions. Such dosage and administration instructions may be the kind provided to a physician, such as through a drug label, or they may be the kind provided by a physician, such as instructions provided to a patient.

[0280] When the combination is administered separately in a sequential manner, wherein one is administered first and then the other, or vice versa, this sequential administration may be close in time or spaced far apart in time. For example, it includes administering another agent a few minutes to tens of minutes after the first agent is administered, and administering another agent a few hours to a few days after the first agent is administered, wherein the time elapsed is not limited. For example, one agent may be administered once a day, while another agent may be administered 2 or 3 times a day, or one agent may be administered once a week, while another agent may be administered once a day, etc. It will be clear to those skilled in the art that, when appropriate, other therapeutic ingredients may be used in the form of salts, such as alkali metal or amine salts or as acid addition salts, or prodrugs, or as esters, such as lower alkyl esters, or as solvates, such as hydrates, to optimize the activity and / or stability and / or physical properties, such as solubility, of the therapeutic ingredients. It will also be clear that, when appropriate, the therapeutic ingredients may be used in optically pure form.

[0281] When combined in the same composition, it will be understood that the two compounds must be stable, compatible with each other and with the other components of the composition, and can be formulated for administration. When formulated separately, they may be provided in any convenient composition, conveniently in a manner known to such compounds in the art.

[0282] When a compound of Formula I is used in combination with a second therapeutic agent active against the same disease, condition or disorder, the dosage of each compound may differ from that when the compound is used alone. Appropriate dosages will be readily appreciated by those skilled in the art.

[0283] In one embodiment, the patient in the methods and uses of the present invention is a mammal. In another embodiment, the patient is a human. The compounds of the present invention can be used to treat diseases and conditions in which modulation of STING is beneficial, including cancer. As modulators of immune responses, the compounds of Formula I and their pharmaceutically acceptable salts can also be used alone, in combination or as an adjuvant to treat diseases and conditions in which modulation of STING is beneficial.

[0284] In one aspect, the disease or condition to be treated is cancer. Examples of cancer diseases and conditions in which a compound of Formula I or a pharmaceutically acceptable salt thereof may have a potentially beneficial anti-tumor effect include cancer of the lung, bone, pancreas, skin, head, neck, uterus, ovary, stomach, colon, breast, esophagus, small intestine, intestine, endocrine system, thyroid, parathyroid, adrenal glands, urethra, prostate, penis, testicles, ureter, bladder, kidney, or liver; rectal cancer; cancer of the anal region; cancer of the fallopian tube, endometrium, cervix, vagina, vulva, renal pelvis, renal cell cancer of the lining of the lining of the lung; cancer of the soft tissue; sarcoma of the soft tissue; myxoma; rhabdomyomas; fibromas; lipomas; teratomas; cholangiocarcinomas; hepatoblastomas; angiosarcomas; hepatomas; fibrosarcomas; chondrosarcomas; myeloma; chronic or acute leukemias; lymphocytic lymphomas; primary CNS lymphomas; CNS tumors; spinal axis tumors; squamous cell carcinomas; synovial sarcomas; malignant pleural mesothelioma; brain stem gliomas; pituitary adenomas; bronchial adenomas; chondromatous hanlartoma; mesothelioma; Hodgkin's disease; or a combination of one or more of the foregoing cancers.

[0285] In another aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof for use in treating cancer.

[0286] In another aspect, the present invention provides a method for treating cancer, which comprises administering a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof.

[0287] In another aspect, the present invention provides use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer.

[0288] The compounds of the invention can be used together with other therapeutic approaches for cancer treatment, for example, in anti-tumor therapy, in combination therapy with immune checkpoint inhibitors, other chemotherapeutic agents, hormones, antibody agents, and surgery and / or radiation therapy.

[0289] Immune checkpoint inhibitors, such as humanized antibodies against PD-1, PD-L1, and CTLA4, have recently been shown to be very successful in treating several types of metastatic cancer, including melanoma, non-small cell lung cancer, renal cell carcinoma, and bladder cancer (Sharma and Allison, 2015, Science 348, 56). However, only a small percentage of cancer patients still benefit from checkpoint inhibitor therapy, in part because of insufficient numbers of anti-tumor immune cells (such as CD8 T cells) generated and / or infiltrated into tumors. Activation of the cGAS-STING pathway activates anti-tumor immunity, including the generation and infiltration of tumor-specific CD8 T cells. Therefore, cGAMP analogs are expected to work synergistically with immune checkpoint inhibitors, and combination therapy may bring therapeutic benefits to a larger percentage of cancer patients.

[0290] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one immune checkpoint inhibitor.

[0291] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one immune checkpoint inhibitor for use in treatment.

[0292] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one immune checkpoint inhibitor for use in treating cancer.

[0293] In another aspect, the present invention provides use of a pharmaceutical composition in the preparation of a medicament for treating cancer, wherein the pharmaceutical composition comprises a compound of formula I or a pharmaceutically acceptable salt thereof and at least one immune checkpoint inhibitor.

[0294] In another aspect, the present invention provides a method for treating cancer, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition, the pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least an immune checkpoint inhibitor.

[0295] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, at least one immune checkpoint inhibitor, and one or more pharmaceutically acceptable carriers, diluents, and excipients.

[0296] Radiation to tumors, especially high-dose radiation, such as stereotactic body radiation therapy (SBRT), kills tumor cells with high precision. Dead tumor cells not only provide tumor antigens to generate tumor-specific cytotoxic T cells, but also release tumor DNA into antigen-presenting cells to activate the cGAS-STING pathway (Deng et al., 2014, Immunity 41, 843). Therefore, cGAMP analogs are expected to work synergistically with radiation therapy to benefit a greater percentage of cancer patients.

[0297] In yet another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, in combination with radiation therapy such as SBRT.

[0298] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, for use in combination with radiation therapy such as SBRT for treating cancer.

[0299] In yet another aspect, the present invention provides use of a pharmaceutical composition for preparing a medicament for treating cancer, the pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, in combination with radiotherapy such as SBRT.

[0300] In another aspect, the present invention provides a method for treating cancer, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition in combination with radiation therapy such as SBRT, wherein the pharmaceutical composition comprises a compound of Formula I or a pharmaceutically acceptable salt thereof.

[0301] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents and excipients, which is combined with radiation therapy SBRT for treating cancer.

[0302] Anti-tumor agents include chemical compounds and antibodies that kill tumor cells by inhibiting the cell cycle, signal transduction, DNA metabolism and angiogenesis and / or by promoting DNA damage, apoptosis and necrosis. These agents comprise the largest class of molecules currently used for cancer treatment. Anti-tumor agents selectively kill tumor cells, although many of them also kill normal cells, resulting in serious side effects. The processing of dead tumor cell-associated antigens by antigen-presenting cells leads to the generation of tumor-specific cytotoxic T cells. This process can be enhanced by cGAMP analogs. Therefore, the combination of cGAMP analogs with anti-tumor agents may produce a synergistic effect, benefiting a greater percentage of patients.

[0303] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof and at least one anti-tumor agent.

[0304] In yet another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof and at least one anti-tumor agent for use in treatment.

[0305] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof and at least one anti-tumor agent for use in treating cancer.

[0306] In another aspect, the present invention provides use of a pharmaceutical composition in the preparation of a medicament for treating cancer, wherein the pharmaceutical composition comprises a compound of formula I or a pharmaceutically acceptable salt thereof and at least one anti-tumor agent.

[0307] In another aspect, the present invention provides a method for treating cancer, comprising: administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one anti-tumor agent.

[0308] In yet another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, at least one anti-tumor agent and one or more pharmaceutically acceptable carriers, diluents and excipients.

[0309] Any antineoplastic agent that is active against the susceptible tumor being treated may be used in combination. Typical antineoplastic agents that may be used include antimicrotubule agents such as diterpenoids and vinca alkaloids; platinum coordination complexes; alkylating agents such as nitrogen mustards, oxazolidinones, alkyl sulfonates, nitrosoureas, and triazenes; antibiotic agents such as anthracyclines, actinomycins, and bleomycins; topoisomerase II inhibitors such as epipodophyllotoxins; antimetabolites such as purine and pyrimidine analogs and antifolates; topoisomerase I inhibitors such as camptothecins; hormones and hormone analogs; signal transduction pathway inhibitors; non-receptor tyrosine angiogenesis inhibitors; immunotherapeutic agents; pro-apoptotic agents; and cell cycle signaling inhibitors.

[0310] Anti-microtubule agents or anti-mitotic agents are cycle-specific agents that are active against the microtubules of tumor cells during the M or mitotic phase of the cell cycle. Examples of anti-microtubule agents include diterpenoids and vinca alkaloids.

[0311] Diterpenoids derived from natural sources are cycle-specific anticancer agents that act in the G2 / M phase of the cell cycle. It is believed that diterpenoids stabilize the β-tubulin subunit of microtubules by binding to this protein. The breakdown of this protein is then inhibited, and mitosis is arrested, followed by cell death. Examples of diterpenoids include paclitaxel and its analog docetaxel.

[0312] Vinca alkaloids are cycle-specific antitumor agents derived from the periwinkle plant. Vinca alkaloids work in the M phase (mitosis) of the cell cycle by specifically binding to tubulin. Therefore, the combined tubulin molecules cannot polymerize into microtubules. It is believed that mitosis stops in metaphase, followed by cell death. The example of vinca alkaloids includes vinblastine, vincristine and vinorelbine.

[0313] Platinum coordination complexes are non-periodic specific anticancer agents that interact with DNA. Platinum complexes enter tumor cells, undergo, hydrate and form intra- and inter-strand crosslinks with DNA, causing adverse biological effects on tumors. Examples of platinum coordination complexes include oxaliplatin, cisplatin and carboplatin.

[0314] Alkylating agents are non-periodic anticancer specific agents and strong electrophiles. Typically, alkylating agents form covalent bonds with DNA through nucleophilic moieties (such as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazole groups) of DNA molecules by alkylation. This alkylation destroys nucleic acid function and causes cell death. Examples of alkylating agents include nitrogen mustards such as cyclophosphamide, melphalan, and chlorambucil; alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; and triazenes such as dacarbazine.

[0315] Antibiotic antineoplastic agents are non-cyclic specific agents that bind to or insert into DNA. Typically, this action results in stable DNA complexes or strand breaks, which disrupt the normal functions of nucleic acids, leading to cell death. Examples of antibiotic antineoplastic agents include actinomycins such as dactinomycin D, anthracyclines such as daunorubicin and doxorubicin; and bleomycin.

[0316] Topoisomerase II inhibitors include epipodophyllotoxins. Epipodophyllotoxins are cycle-specific antitumor agents derived from the Datura plant. Epipodophyllotoxins typically affect cells in the S and G2 phases of the cell cycle by forming a ternary complex with topoisomerase II and DNA, resulting in DNA strand breaks. Strand breaks accumulate, followed by cell death. Examples of epipodophyllotoxins include etoposide and teniposide.

[0317] Antimetabolite antitumor agents are cycle-specific antitumor agents that act in the S phase (DNA synthesis) of the cell cycle by inhibiting DNA synthesis or by inhibiting purine or pyrimidine base synthesis to limit DNA synthesis. Therefore, the S phase no longer proceeds, and the cell dies subsequently. Examples of antimetabolite antitumor agents include fluorouracil, methotrexate, cytarabine, mercaptopurine, thioguanine, and gemcitabine.

[0318] Camptothecins, including camptothecin and camptothecin derivatives, are available or under development as inhibitors of topoisomerase I. It is believed that the cytotoxic activity of camptothecins is related to their topoisomerase I inhibitory activity. Examples of camptothecins include, but are not limited to, irinotecan, topotecan, and various optical forms of 7-(4-methylpiperazine-methylene)-10,11-ethylenedioxy-20-camptothecin described below.

[0319] Hormones and hormone analogs are compounds that can be used to treat cancer where there is a relationship between the hormone and the growth and / or lack of growth of the cancer. Examples of hormones and hormone analogs that can be used to treat cancer include adrenocortical hormones such as prednisone and prednisolone, which are used to treat malignant lymphomas and acute leukemias in children; aminoglutethimide and other aromatase inhibitors such as anastrozole, letrazole, vorazole, and exemestane, which are used to treat adrenocortical carcinoma and hormone-dependent breast cancers that contain estrogen receptors; progestins such as megestrol acetate, which are used to treat hormone-dependent breast cancer and endometrial cancer; estrogens and anti-estrogens such as fulvestrant, flutamide, nilutamide, bicalutamide, cyproterone acetate, and valproate. Progesterone and 5a-reductase such as finasteride and dutasteride, which are used to treat prostate cancer and benign prostatic hypertrophy; antiestrogens such as tamoxifen, toremifene, raloxifene, droloxifene, iodoxyfene, and selective estrogen receptor modulators (SERMS), which are used to treat hormone-dependent breast cancer and other susceptible cancers; and gonadotropin-releasing hormone (GnRH) and its analogs, which stimulate the release of luteinizing hormone (LH) and / or follicle-stimulating hormone (FSH) for the treatment of prostate cancer, for example, LHRH agonists and antagonists such as goserelin acetate and luprolide.

[0320] Signal transduction pathway inhibitors are those inhibitors that block or inhibit the chemical processes that cause changes in cells. As used herein, this change is cell proliferation or differentiation. Signal transduction inhibitors that can be used in the present invention include the following inhibitors: receptor tyrosine kinases, non-receptor tyrosine kinases, SH2 / SH3 domain blockers, serine / threonine kinases, phosphatidylinositol-3 kinases, inositol signaling and Ras oncogenes.

[0321] Several protein tyrosine kinases catalyze the phosphorylation of specific tyrosyl residues in various proteins involved in the regulation of cell growth. These protein tyrosine kinases can be broadly classified as either receptor or non-receptor kinases.

[0322] Receptor tyrosine kinases are transmembrane proteins with an extracellular ligand binding domain, a transmembrane domain, and a tyrosine kinase domain. Receptor tyrosine kinases are involved in the regulation of cell growth and are often referred to as growth factor receptors. Inappropriate or uncontrolled activation of many of these kinases, i.e., aberrant kinase growth factor receptor activity, e.g., by overexpression or mutation, has been shown to lead to uncontrolled cell growth. Thus, aberrant activity of these kinases is associated with malignant tissue growth. Therefore, inhibitors of these kinases may provide a cancer therapy approach. Growth factor receptors include, for example, epidermal growth factor receptor (EGFr), platelet-derived growth factor receptor (PDGFr), erbB2, erbB4, ret, vascular endothelial growth factor receptor (VEGFr), tyrosine kinase with immunoglobulin-like and epidermal growth factor homology domains (TIE-2), insulin growth factor-I (IGFI) receptor, macrophage colony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growth factor (FGF) receptor, Trk receptor (TrkA, TrkB and TrkC), ephrin (eph) receptor and RET proto-oncogene. Several growth receptor inhibitors are under development, including ligand antagonists, antibodies, tyrosine kinase inhibitors and antisense oligonucleotides. Growth factor receptors and agents that inhibit growth factor receptor function are described, for example, in Kath, John C, Exp. Opin. Ther. Patents (2000) 10(6):803-818; Shawver et al. DDT Vol 2, No. 2, February 1997; and Lofts, FJ et al. "Growth factor receptors as targets", New Molecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr, David, CRC Press 1994, London.

[0323] Tyrosine kinases that are not growth factor receptor kinases are called non-receptor tyrosine kinases. Non-receptor tyrosine kinases that can be used in the present invention are targets or potential targets of anticancer drugs, including cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (focal adhesion kinase), Brutons tyrosine kinase and Bcr-Abl. Such non-receptor kinases and agents that inhibit the function of non-receptor tyrosine kinases are described in, for example, Sinh, S. and Corey, SJ, (1999) Journal of Hematotherapy and Stem Cell Research 8 (5): 465–80; and Bolen, JB, Brugge, JS, (1997) Annual review of Immunology. 15: 371-404.

[0324] SH2 / SH3 domain blockers are agents that disrupt SH2 or SH3 domain binding in a variety of enzymes or adaptor proteins, including PI3-K p85 subunit, Src family kinases, adaptor molecules (She, Crk, Nek, Grb2) and Ras-GAP. The SH2 / SH3 domain as a target for anticancer drugs is discussed in the following literature: Smithgall, TE (1995), Journal of Pharmacological and Toxicological Methods. 34 (3) 125-32.

[0325] Inhibitors of serine / threonine kinases include MAP kinase cascade blockers, including blockers of Raf kinase (rafk), mitogen- or extracellular-regulated kinase (MEK) and extracellular-regulated kinase (ERK); and protein kinase C family member blockers, including blockers of PKC (α, β, γ, ε, μ, λ, ι, ζ), I kB kinase family (I KKa, I KKb), PKB family kinases, akt kinase family members and TGFβ receptor kinase. These serine / threonine kinases and their inhibitors are described, for example, in Yamamoto, T. et al., (1999), Journal of Biochemistry. 126 (5) 799-803; Brodt, P et al. (2000), Biochemical Pharmacology, 60. 1101-1107; Massague, J., and Weis-Garcia, F. (1996) Cancer Surveys. 27: 41-64; Philip, P. A. and Harris, A. L. (1995), Cancer Treatment and Research. 78: 3-27; Lackey, K. et al Bioorganic and Medicinal Chemistry Letters, (10), 2000, 223-226; U.S. Pat. No. 6,268,391; and Martinez-lacaci, L, et al., Int. J. Cancer (2000), 88 (1), 44-52.

[0326] Inhibitors of members of the phosphatidylinositol-3 kinase family, including blockers of PI3-kinase, ATM, DNA-PK and Ku, can also be used in the present invention. These kinases are discussed, for example, in the following literature: Abraham, RT (1996), Current Opinion in Immunology. 8 (3) 412-8; Canman, CE, Lim, DS (1998), Oncogene 17 (25) 3301-3308; Jackson, SP (1997), International Journal of Biochemistry and Cell Biology. 29 (7): 935-8; and Zhong, H. et al. Cancer Res., (2000) 60 (6), 1541-1545.

[0327] Also useful in the present invention are Myo-inositol signaling inhibitors, such as phospholipase C blockers and Myo-inositol analogs. Such signaling inhibitors are described, for example, in Powis, G., and Kozikowski A., (1994) New Molecular Targets for Cancer Chemotherapy, ed. Paul Workman and David Kerr, CRC Press 1994, London.

[0328] Another group of signal transduction pathway inhibitors are inhibitors of Ras oncogenes. Such inhibitors include inhibitors of farnesyltransferase, geranyl-geranyltransferase, and CAAX proteases, as well as antisense oligonucleotides, ribozymes, and immunotherapy. Such inhibitors have been shown to block ras activation in cells containing wild-type mutant ras, thereby acting as antiproliferative agents. Ras oncogene inhibition is discussed in the following literature: Scharovsky, OG, et al. (2000), Journal of Biomedical Science. 7 (4) 292-8; Ashby, MN (1998), Current Opinion in Lipidology. 9 (2) 99–102; and BioChim. Biophys. Acta, (19899) 1423 (3): 19-30.

[0329] Receptor kinase ligand binding antibody antagonists can also be used as signal transduction inhibitors. This group of signal transduction pathway inhibitors includes the use of humanized antibodies for the extracellular ligand binding domain of receptor tyrosine kinases. Examples include Imclone C225 EGFR-specific antibodies (see Green, MC et al, Monoclonal Antibody Therapy for Solid Tumors, Cancer Treat. Rev., (2000), 26 (4), 269-286); erbB2 antibody (see Tyrosine Kinase Signalling in Breast cancer: erbB Family Receptor Tyrosine Kinases, Breast cancer Res., 2000, 2(3), 176-183); and 2CB VEGFR2-specific antibody (see Brekken, RA et al, Selective Inhibition of VEGFR2 Activity by a monoclonal Anti-VEGF antibody blocks tumor growth in mice, Cancer Res. (2000) 60, 5117-5124).

[0330] Antiangiogenic agents such as non-receptor MEK angiogenesis inhibitors may also be useful, as well as those that inhibit the action of vascular endothelial growth factor (e.g., the anti-vascular endothelial growth factor antibody bevacizumab [Avastin TM ]) and compounds that act through other mechanisms (e.g., linoamide, inhibitors of integrin αvβ3 function, endostatin and angiostatin).

[0331] In one aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, and at least one anti-tumor agent, which anti-tumor agent is an anti-microtubule agent, a platinum coordination complex, an alkylating agent, an antibiotic agent, a topoisomerase II inhibitor, an antimetabolite, a topoisomerase I inhibitor, a hormone and hormone analog, a signal transduction pathway inhibitor, a non-receptor tyrosine MEK angiogenesis inhibitor, an immunotherapeutic agent, a pro-apoptotic agent, or a cell cycle signaling inhibitor.

[0332] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and at least one antitumor agent selected from diterpenoids and vinca alkaloids.

[0333] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and at least one anti-tumor agent, which is a platinum coordination complex. In one embodiment, at least one anti-tumor agent is paclitaxel, carboplatin or vinorelbine.

[0334] On the other hand, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and at least one antitumor agent, which is a signal transduction pathway inhibitor. In one embodiment, the signal transduction pathway inhibitor is an inhibitor of growth factor receptor kinases VEGFR2, TIE2, PDGFR, BTK, erbB2, EGFr, IGFR-1, TrkA, TrkB, TrkC or c-fms. In another embodiment, the signal transduction pathway inhibitor is an inhibitor of serine / threonine kinases rafk, akt or PKC-ζ. In another embodiment, the signal transduction pathway inhibitor is an inhibitor of non-receptor tyrosine kinases selected from the src kinase family. In another embodiment, the signal transduction pathway inhibitor is an inhibitor of c-src. In another embodiment, the signal transduction pathway inhibitor is an inhibitor of Ras oncogenes, which is selected from inhibitors of farnesyl transferase and geranylgeranyl transferase. In another embodiment, the signal transduction pathway inhibitor is an inhibitor of serine / threonine kinases selected from PI3K. In another embodiment, the signal transduction pathway inhibitor is a dual EGFr / erbB2 inhibitor, such as N-{3-chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methylsulfonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamine.

[0335] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and at least one anti-tumor agent, which is a cell cycle signaling inhibitor. In one embodiment, the cell cycle signaling inhibitor is an inhibitor of CDK2, CDK4 or CDK6.

[0336] The compound of formula I can be prepared by methods known in the field of organic synthesis, as described in the scheme below and / or in the specific embodiments described below. In all these methods, it is fully understood that, according to the general principles of chemistry, protecting groups can be used for sensitive or reactive groups if necessary. Protective groups (TW Green and PGM Wuts (1999) Protective Groups in Organic Synthesis, the third edition, John Wiley & Sons) are manipulated according to standard methods for organic synthesis. These groups are removed at the appropriate stage of compound synthesis using methods that are obvious to those skilled in the art. The selection of method and reaction conditions and their order of implementation should be consistent with the preparation of the compound of formula I.

[0337] It should be understood that the examples and embodiments described herein are for illustrative purposes only and will suggest various modifications or changes thereto that may occur to those skilled in the art, and that these modifications or changes are included within the spirit and purview of this application and the scope of the appended claims.

[0338] The following list provides definitions of certain abbreviations used herein. It should be understood that this list is not exhaustive, but the meanings of those abbreviations not defined below will be apparent to those skilled in the art: Ac is acetyl; AcOH is acetic acid; Ac2O is acetic anhydride; AIBN is 2,2'-azobisisobutyronitrile; Bn is benzyl; BSA is N,O-bis(trimethylsilyl)acetamide; BSTFA is N,O-bis(trimethylsilyl)trifluoroacetamide; Bu is butyl; Bz is benzoyl; CAN is cerium ammonium nitrate; CE is 2-cyanoethyl; DCA is dichloroacetic acid; DCM is dichloromethane; DDTT is 1,2,4-dithiazole-5-thione; DEAD is diethyl azodicarboxylate; DIAD is diisopropyl azodicarboxylate; DIPEA is N,N-diisopropylethylamine; DMAP is 4-(dimethylamino)pyridine; DMF is N,N-dimethylformamide; DMOCP is 2-chloro-5,5-dimethyl-1,3,2-dioxaphosphorinane 2-oxide; DMSO is dimethyl sulfoxide; DMTr is 4,4'-dimethoxytrityl; EtOAc is ethyl acetate; EtOH is ethanol; HMPT is hexamethylphosphorotriamide; HPLC is high performance liquid chromatography; ibu is isobutyryl; IBX is 2-iodoacylbenzoic acid; Imid is imidazole; i Pr is isopropyl; KOH is potassium hydroxide; Me is methyl; MeCN is acetonitrile; MeOH is methanol; MTBE is methyl tert-butyl ether; Ms is methanesulfonyl; Pd / C is palladium on activated carbon; NIS is N-iodosuccinimide; NPE is 2-(4-nitrophenyl)ethyl; PE is petroleum ether; Ph is phenyl; PMB is p-methoxybenzyl; PPh3 is triphenylphosphine; Py is pyridine; TBAF is tetra-n-butylammonium fluoride; TBAI is tetrabutylammonium iodide; TBDPS is tert-butyldiphenylsilyl; TBHP is tert-butyl TBS is tert-butyldimethylsilyl; TCDI is 1,1'-thiocarbonyldiimidazole, TDA-1 is tris[2-(2-methoxyethoxy)ethyl]amine; TEA is triethylamine; Tf is trifluoromethanesulfonyl; TFA is trifluoroacetic acid; TFE is 2,2,2-trifluoroethyl; THF is tetrahydrofuran; TIPS is triisopropylsilyl; TLC is thin layer chromatography; TMS is trimethylsilyl: TMSOTf is trimethylsilyl trifluoromethanesulfonate; Tol is p-toluoyl; Tr is trityl.

[0339] Intermediate preparation

[0340]

[0341] Step 1: Ketal 2

[0342]

[0343] To a suspension of D-ribose (1) (160 g, 1.07 mol) in acetone (2.0 L) was added concentrated sulfuric acid (10.7 g, 107 mmol, 5.8 mL) dropwise at 27°C. After stirring for 12 hours, solid sodium bicarbonate (100 g) was added. The mixture was then filtered, and the filtrate was concentrated to give crude compound 2 (215.0 g).

[0344] Step 2: Silyl ether 3

[0345]

[0346] To a solution of crude compound 2 (215 g, 1.13 mol) in DCM (1.5 L) was added TBSCl (170 g, 1.13 mol) and TEA (172 g, 1.69 mol) at 0° C. After stirring at 27° C. for 12 hours, the mixture was filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 100 to 1 / 50) to give 3 as a colorless oil (285 g, 83% yield).

[0347] Step 3: Deazapurine 4

[0348]

[0349] To a solution of 3 (60.0 g, 197.08 mmol) and carbon tetrachloride (67.3 g, 438 mmol, 42 mL) in THF (1.2 L) was added HMPT (63.0 g, 386 mmol, 70 mL) dropwise at -78°C and stirred at 27°C for 2 hours. To another solution of 6-chloro-7-deazapurine (24.2 g, 158 mmol) and KOH (16.6 g, 296 mmol) in MeCN (1.2 L) was added TDA-1 (6.37 g, 19.7 mmol) at 27°C, followed by the THF solution obtained above. After stirring at 27°C for 12 hours, the reaction mixture was filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 50 to 1 / 10) to give 4 as a yellow oil (15.3 g, 18% yield).

[0350] Step 4: Adenine 5

[0351]

[0352] A solution of 4 (28.6 g, 64.9 mmol) in dioxane (150 mL) and aqueous ammonium hydroxide solution (500 mL) was stirred for 30 hours at 120 ° C in a sealed autoclave. The volatiles were then removed and the aqueous solution was extracted with EA (300 mL × 3). The combined organic layer was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and partially purified by silica gel column chromatography (EA / DCM=1 / 1) to give a yellow foam (9.65 g). The residue was then dissolved in THF (50 mL) and treated with TBAF trihydrate (10.9 g, 34.4 mmol) at 27 ° C. After stirring for 2 hours, the mixture was concentrated and purified by silica gel column chromatography (EA / DCM=1 / 1 to 7 / 1) to give 5 (5.56 g, 79% yield) as a yellow solid.

[0353] Step 5: Benzoate 6

[0354]

[0355] To a solution of 5 (7.26 g, 23.7 mmol) in DCM (60 mL) was added Imid (4.84 g, 71.1 mmol) and TBSCl (5.36 g, 35.6 mmol) at 27°C. After stirring at 27°C for 1.5 hours, water (100 mL) was added, and the mixture was extracted with DCM (200 mL). The organic layer was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give crude TBS-5. Benzoyl chloride (5.14 g, 36.6 mmol) was added to the solution of the crude TBS-5 obtained above in DCM (100 mL) at 27°C. After stirring for 12 hours, water (200 mL) was added, and the mixture was extracted with DCM (500 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE = 1 / 50 to 1 / 10) to give 6 as a yellow foam (8.12 g, 64% yield).

[0356] Step 6: B1

[0357]

[0358] A solution of 6 (15.2 g, 28.9 mmol) in TFA (90 mL) and DCM (20 mL) was stirred for 12 h at 27° C. The volatiles were then removed and the residue was purified by silica gel column chromatography (MeOH / DCM=1 / 100 to 1 / 10) to give B1 as a yellow solid (10.16 g, 95% yield).

[0359]

[0360] Step 1: Aniline 8

[0361]

[0362] To a solution of 5-amino-4,6-dichloropyrimidine (63.0 g, 384 mmol) in n-BuOH (300.0 mL) was added p-methoxybenzylamine (58.0 g, 423 mmol, 55 mL) and DIPEA (99.3 g, 768 mmol, 134 mL). After stirring at 100–110 °C for 15 h, the volatiles were removed and MTBE (100 mL) was added. The solid was collected by filtration and washed with EA to give 8 as an off-white solid (55.0 g, 54% yield). (MS: [M+H] + 265.0)

[0363] Step 2: Azapurine 9

[0364]

[0365] To a solution of 8 (10.0 g, 37.8 mmol) in a mixture of DCM (200 mL), AcOH (100 mL) and water (100 mL) at 0°C was added sodium nitrite (2.87 g, 41.6 mmol, 2.3 mL). After stirring at 0–25°C for 1 hour, DCM (30 mL) and saturated aqueous sodium bicarbonate solution (30 mL) were added. The layers were then separated, and the aqueous phase was extracted with DCM (150 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 3) to give 9 as a light yellow solid (6.0 g, 88% yield). (MS: [M+H] + 276.0)

[0366] Step 3: Azaadenine 10

[0367]

[0368] To a solution of 9 (6.0 g, 21.8 mmol) in 1,4-dioxane (30 mL) was added aqueous ammonium hydroxide (30 mL). After stirring at 30-40 °C for 5 h, the solid was collected by filtration to afford 10 as a white solid (4.0 g, 70% yield). (MS: [M+H] + 257.1)

[0369] Step 4: Azaadenine 11

[0370]

[0371] To a solution of 10 (17.0 g, 66.3 mmol) in Py (100 mL) was added DMAP (8.92 g, 73.0 mmol), Imid (13.6 g, 199 mmol) and benzoyl chloride (14.0 g, 99.5 mmol, 11.6 mL). After stirring at 110-120 °C for 18 hours, the volatiles were removed and DCM (300 mL) and water (300 mL) were added. The layers were separated, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1:1) to give 11 as an off-white solid (17.0 g, 68% yield). (MS: [M+H] + 361.2)

[0372] Step 5: Azaadenine 12

[0373]

[0374] To a suspension of 11 (6.40 g, 17.8 mmol) in MeCN (60 mL) was added a solution of CAN (29.2 g, 53.3 mmol) and sodium bicarbonate (1.49 g, 17.76 mmol) in water (60 mL) at 0 °C. After stirring at 0-25 °C for 12 h, the mixture was neutralized to ~pH 7 with sodium bicarbonate. The solid was collected by filtration to give 12 (2.6 g, 57% yield). (MS: [M+H] + 241.1)

[0375] Step 6: Azaadenosine 14

[0376]

[0377] Tin (IV) chloride (30.3 g, 116 mmol, 13.6 mL) was added to a solution of 12 (9.30 g, 38.7 mmol) and 13 (20.5 g, 40.7 mmol) in MeCN (350 mL) at 0°C. After stirring at 0–25°C for 24 h, the reaction mixture was poured into a saturated aqueous sodium bicarbonate solution (300 mL). The solid was filtered off and washed with water (100 mL). The filtrate was extracted with DCM (150 mL×4), and the combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / DCM=1 / 10) to give 14 as an off-white gum (6.10 g, 21% yield). (MS: [M+H] + 684.9)

[0378] Step 7: B2

[0379]

[0380] To a solution of 14 (6.1 g, 8.9 mmol) in a mixture of THF (35 mL) and MeOH (28 mL) was added aqueous lithium hydroxide solution (1 M, 16.0 mL) at 0°C. After stirring at 0–25°C for 3 h, the mixture was neutralized to ~pH 7 with aqueous citric acid solution (1 M), then concentrated and purified by silica gel column chromatography (MeOH / DCM = 1 / 20) to give B2 as an off-white solid (2.9 g, 87% yield). (MS: [M+H] + 373.1)

[0381]

[0382] Step 1: Diol 16

[0383]

[0384] To a solution of 15 (880 mg, 2.5 mmol) in Py (10 mL) was added a solution of DMTrCl (940 mg, 2.6 mmol) in Py (5 mL). After stirring for 3 hours, the mixture was concentrated and purified by silica gel column chromatography (MeOH / DCM=1 / 20 to 1 / 10) to give 16 as a white foam (1.23 g, 75% yield). (MS: [M+H] + 656.2)

[0385] Step 2: Alcohol 17

[0386]

[0387] To a solution of 16 (900 mg, 1.4 mmol) and Imid (280 mg, 4.15 mmol) in Py (15 mL) was added TBSCl (310 mg, 2.05 mmol). After stirring for 4 hours, the volatiles were removed and the residue was dissolved in DCM (50 mL), washed with saturated aqueous sodium bicarbonate solution and brine, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (EA / toluene=1 / 3 to 2 / 3) to give 17 as a white solid (480 mg, 45% yield). (MS: [M+H] + 770.2)

[0388] Step 3: Thiocarbamate 18

[0389]

[0390] To a solution of 17 (500 mg, 0.65 mmol) in DMF (6 mL) was added TCDI (350 mg, 1.94 mmol). After stirring for 2 days, EA (40 mL) and water (25 mL) were added, and the layers were separated. The aqueous layer was extracted with ethyl acetate (25 mL×3). The combined organic layers were washed with water (20 mL), brine (20 mL×2), dried over anhydrous sodium sulfate, and concentrated to give crude compound 18. (MS: [M+H] + 880.2)

[0391] Step 4: Silyl ether 19

[0392]

[0393] A degassed solution of crude compound 18 in toluene (10 mL) at 110° C. was added with AIBN (57 mg, 0.34 mmol), tributyltin hydride (0.51 mL, 1.94 mmol) in toluene (3 mL) over 30 minutes. After stirring at 110° C. for 6 hours, the mixture was cooled to room temperature, concentrated, and purified by silica gel column chromatography (EA / hexane=1 / 5 to 2 / 1) to give 19 as a yellow oil (195 mg, 40% yield for two steps). (MS: [M+H] + 754.2)

[0394] Step 5: DMTr-B3

[0395]

[0396] To a solution of 19 (190 mg, 0.252 mmol) in THF (5 mL) was added TBAF (1 M in THF, 0.50 mL). After stirring at room temperature for 2 hours, water (5 mL) was added, and the mixture was extracted with EA (8 mL×3), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (MeOH / DCM=1 / 20) to give DMTr-B3 as a white solid (132 mg, 82% yield). (MS: [M+H] + 640.2)

[0397]

[0398] Step 1: Alcohol 21

[0399]

[0400] To a solution of 20 (12.8 g, 67.0 mmol) in Py (300 mL) was added TBDPSCl (21.0 mL, 80.4 mmol). After stirring for 3 h, MeOH (25 mL) was added and the mixture was concentrated. The residue was dissolved in ether (200 mL), washed with aqueous sodium bicarbonate solution (10%, 100 mL) and water (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (ether / PE=1 / 2) to give 21 as a white solid (27.2 g, 95% yield). (MS: [M+Na] + 451.2)

[0401] Step 2: Alcohol 22

[0402]

[0403] A solution of 21 (27.2 g, 63.7 mmol) in DMSO (200 mL) and Ac2O (50 mL) was stirred for 16 hours and then poured into ice water (200 mL). The mixture was extracted with ether (100 mL×3), and the combined organic layers were washed with aqueous sodium bicarbonate solution (10%, 100 mL) and water (100 mL), and concentrated. The residue was then dissolved in MeOH (250 mL) and DCM (250 mL) at 0°C, followed by the addition of sodium borohydride (12.0 g) in 10 portions. After stirring for 5 minutes, water (100 mL) was added and the layers were separated. The organic layer was then concentrated and purified by silica gel column chromatography (ether / PE=1 / 2) to give 22 as a white solid (20.4 g, 75% yield for two steps). (MS: [M+Na] + 451.2)

[0404] Step 3: Methyl ether 23

[0405]

[0406] To a solution of 22 (4.0 g, 9.33 mmol) in DMF (45 mL) at 0°C was added sodium hydride (484 mg, 12.1 mmol) and stirred for 30 minutes, then iodomethane (0.64 mL, 10.3 mmol) was slowly added. After stirring for 3 hours, water (3 mL) was added, and the volatiles were removed and purified by silica gel column chromatography (EA / PE=1 / 10) to give 23 as a white solid (3.8 g, 92% yield). (MS: [M+Na] + 465.2)

[0407] Step 4: Benzoate 24

[0408]

[0409] To a solution of 23 (3.1 g, 7.0 mmol) in THF (50 mL) was added TBAF (8.4 mL, 8.4 mmol) at 0 ° C. After stirring at room temperature for 4 h, water (5 mL) and EA were added. The layers were separated, and the organic layer was washed with water and brine, concentrated, and the resulting residue was dissolved in DCM, followed by the addition of TEA (4.9 mL, 35 mmol) and benzoyl chloride (0.98 mL, 8.4 mmol). After stirring for 1 hour, water (3 mL) was added, and the volatiles were removed. The residue was purified by silica gel column chromatography (EA / PE=1 / 5) to give 24 as a white solid (1.9 g, 88% yield). (MS: [M+Na] + 331.0)

[0410] Step 5: Acetate 25

[0411]

[0412] A solution of 24 (0.71 g, 2.3 mmol) in HOAc (14 mL) and water (6 mL) was heated at reflux for 30 minutes. After cooling to room temperature, the mixture was co-evaporated with toluene (10 mL×4), and the resulting residue was then dissolved in Py / Ac2O (10 / 1 v / v, 10 mL), followed by the addition of DMAP (50 mg, 0.46 mmol). After stirring for 4 hours, the mixture was concentrated and purified by silica gel column chromatography (EA / PE=1 / 3) to give 25 as a white solid (0.75 g, 92% yield). (MS: [M+Na] + 375.0)

[0413] Step 6: Guanosine 27

[0414]

[0415] 25 (500 mg, 1.42 mmol) and N were added to 80 °C. 2 - A suspension of isobutyrylguanine (500 mg, 2.13 mmol) in DCM (20 mL) was added with BSA (1.8 mL, 7.4 mmol) and stirred for 1 hour, and then TMSOTf (0.77 mL, 4.26 mmol) was added. After stirring at 80 ° C for 3 hours, the mixture was cooled to room temperature, and then an aqueous sodium bicarbonate solution (50 mL) was added. The mixture was then extracted with DCM (50 mL×3). The combined organic layer was dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (MeOH / DCM=1 / 20 to 1 / 10) to give 26 as a white powder (624 mg, 85% yield). (MS: [M+H] + 514.2)

[0416] Step 7: B4

[0417]

[0418] To a solution of 27 (0.49 g, 0.96 mmol) in MeOH / THF / water (4 / 5 / 1 v / v / v, 20 mL) at 0°C was added aqueous sodium hydroxide solution (10 M, 0.25 mL, 2.5 mmol). After stirring for 30 minutes, HOAc was added, and the mixture was concentrated and purified by silica gel column chromatography (MeOH / DCM=1 / 10 to 1 / 5) to give B4 as an oil (322 mg, 92% yield). (MS: [M+H] + 368.2)

[0419]

[0420] Step 1: Acetate 28

[0421]

[0422] To a solution of 15 (7.0 g, 20 mmol) in MeCN (100 mL) was added DMAP (1.2 g, 10 mmol) and Ac2O (7.5 mL, 80 mmol) at 0°C. After stirring overnight at room temperature, the mixture was concentrated and purified by silica gel flash chromatography (MeOH / DCM = 1 / 20 to 1 / 10) to give 27 as a white solid (8.77 g, 92% yield). (MS: [M+H] + 480.0)

[0423] Step 2: Propargyl ether 29

[0424]

[0425] To a solution of 27 (480 mg, 1.0 mmol) in 1,4-dioxane (1 mL) was added PPh3 (656 mg, 2.5 mmol), propargyl bromide (0.15 mL, 2 mmol) and DEAD (0.49 mL, 2.5 mmol) in dioxane (1 mL) at 0°C. After stirring for 2 hours, the mixture was concentrated and purified by silica gel column chromatography (MeOH / DCM=1 / 50 to 1 / 20) to give 28 as a white solid (440 mg, 47% yield). (MS: [M+H] + 518.2)

[0426] Step 3: B5

[0427]

[0428] To a solution of 28 (150 mg, 0.17 mmol) in THF (4.5 mL) and MeOH (0.5 mL) was added aqueous sodium hydroxide solution (1 M, 0.5 mL) at 0°C. After stirring for 1 hour, HOAc (0.1 mL) was added, and the mixture was concentrated and purified by silica gel column chromatography (MeOH / DCM=1 / 20 to 1 / 10) to give B5 as a white solid (40 mg, 64% yield). (MS: [M+H] + 392.0)

[0429]

[0430] Step 1: Deazapurine 29

[0431]

[0432] To a solution of 3 (40.0 g, 131 mmol) and carbon tetrachloride (33.6 g, 218 mmol, 21 mL) in THF (500 mL) at -78 °C was added HMPT (22.5 g, 138 mmol, 25 mL) over 15 min. After stirring for 2 hours and gently warming for a short time to prevent gel formation, the mixture was concentrated to about 70 mL. To a suspension of KOH (25.8 g, 460 mmol) in MeCN (600 mL) was added TDA-1 (4.25 g, 13.14 mmol, 4.2 mL). After stirring for 10 minutes at 25 °C, 2-amino-6-chloro-7-deazapurine (22.2 g, 131 mmol) was added. The mixture was stirred for another 10 minutes, and then the THF solution obtained above was added. After stirring for 2 hours, the mixture was filtered, concentrated, and purified by silica gel column chromatography (EA / PE=3 / 17) to give 29 (9.20 g, 15% yield). (MS: [M+H] + 455.3)

[0433] Step 2: Alcohol 30

[0434]

[0435] To a mixture of 29 (13.7 g, 30.1 mmol) in dioxane (10 mL) was added a solution of sodium hydroxide (11.7 g, 291 mmol) in water (100 mL) at 25 °C. After stirring at 80 °C for 64 hours, the mixture was cooled to 0 °C, neutralized to ~pH 7 with AcOH, and extracted with EtOAc (100 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated to give crude compound 30. (MS: [M+1] + 323.1)

[0436] Step 3: Silyl ether 31

[0437]

[0438] To a solution of crude compound 30 (9.7 g, 30.1 mmol) and Imid (4.1 g, 60.3 mmol) in DCM (10 mL) was added TBSCl (9.08 g, 60.3 mmol) at 25°C. After stirring for 16 hours, the mixture was diluted with DCM (100 mL), washed with brine (80 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 4 to 1 / 1, then MeOH / DCM=1 / 50) to give 31 (9.0 g, 68% yield) as a solid. (MS: [M+H] + 437.2)

[0439] Step 4: Isobutyrate 32

[0440]

[0441] To a solution of 31 (9.0 g, 20.6 mmol) and TEA (4.2 g, 41.2 mmol) in DCM (80 mL) was added isobutyryl chloride (3.29 g, 30.9 mmol) at 0°C. After stirring at 25°C for 16 hours, the mixture was diluted with DCM (100 mL), washed with saturated aqueous sodium bicarbonate solution (50 mL×2) and brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 5 to 1 / 2) to give 32 as a white solid (4.2 g, 40% yield). (MS: [M+H] + 507.2)

[0442] Step 5: B6

[0443]

[0444] A solution of 32 (4.2 g, 8.29 mmol) in DCM (6 mL) and TFA (24 mL) was stirred at 25 °C for 1 hour before concentration. The residue was then treated with hydrogen chloride (4M in MeOH, 10 mL) at 0 °C. After stirring at 25 °C for 10 minutes, the mixture was concentrated to give crude compound B6 as a white solid (2.92 g, 99% yield). (MS: [M+H] + 353.0)

[0445]

[0446] Step 1: Propargyl ether 34

[0447]

[0448] To a solution of adenosine (33) (5.0 g, 18.7 mmol) in DMF (200 mL) at 0° C. was added sodium hydride (60% dispersion in mineral oil, 1.0 g, 25 mmol), followed by TBAI (1.5 g, 4.06 mmol) and propargyl bromide (2.12 mL, 20.9 mmol). After stirring at 55° C. for 2 days, the mixture was purified by silica gel column chromatography (MeOH / DCM=7 / 93) and then recrystallized from ethanol to give 34 as a pale yellow solid (2.56 g, 45%).

[0449] Step 2: BA1

[0450]

[0451] To a solution of 34 (1.4 g, 4.59 mmol, co-evaporated twice with Py) in Py (20 mL) was added TMSCl (2.4 mL, 18.9 mmol). After stirring for 30 minutes, benzoyl chloride (0.7 mL, 6.0 mmol) was added, and the mixture was stirred for 3 hours, and then water (10 mL) and aqueous ammonium hydroxide solution (15 mL) were added at 0 ° C. After stirring at room temperature for 20 minutes, the mixture was extracted with DCM (25 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (MeOH / DCM=5:95) to give BA1 as a white foam (1.73 g, 92%). (MS: [M+H] + 410.2)

[0452]

[0453] Step 1: Pyrazolopyrimidine 36

[0454]

[0455] To a solution of 35 (10.0 g, 56.5 mmol) in THF (80 mL) was added DIPEA (7.3 g, 56.5 mmol, 9.9 mL). After stirring at 0 ° C for 10 minutes, a solution of hydrazine (1.81 g, 56.5 mmol, 2.0 mL) in THF (20 mL) was added. The mixture was then stirred at 20 ° C for 2 hours and then concentrated. After adding DCM (100 mL) and H2O (100 mL) to the residue, the layers were separated and the aqueous layer was extracted with DCM (100 mL×3). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 15 to 1 / 8) to give 36 as a yellow solid (3.10 g, 35% yield). (MS: [M+H] + 155.1)

[0456] Step 2: Pyrazolopyrimidine 37

[0457]

[0458] To a solution of 36 (200 mg, 1.29 mmol) in THF (2.0 mL) was added ammonium hydroxide (2.0 mL). After stirring at 20-30 °C for 2 h, the mixture was concentrated, triturated with MeCN (0.5 mL), and collected by filtration to give 37 as a red solid (100 mg, 57% yield).

[0459] Step 3: Tribenzoate 38

[0460]

[0461] To a suspension of 37 (20.0 g, 148 mmol) and 13 (101 g, 200 mmol) in MeCN (1.2 L) was added boron trifluoride etherate (30.5 g, 215 mmol, 26.5 mL). After stirring at 75-85 ° C for 2 hours, the mixture was concentrated and purified by silica gel column chromatography (EA / PE=1 / 5 to 2 / 1) to give 38 as a yellow solid (35.0 g, 40% yield). (MS: [M+H] + 580.3)

[0462] Step 4: Benzamide 39

[0463]

[0464] To a solution of 38 (10.0 g, 17.3 mmol) in DCM (100 mL) was added DMAP (421 mg, 3.45 mmol) and TEA (5.24 g, 51.8 mmol, 7.2 mL) dropwise, followed by the addition of benzoyl chloride (2.91 g, 20.7 mmol, 2.4 mL) dropwise. After stirring at 20-25 °C for 8 hours, the mixture was concentrated and purified by silica gel column chromatography (EA / PE=1 / 5 to 11 / ) to give 39 as a white solid (9.0 g, 76% yield). (MS: [M+H] + 684.1)

[0465] Step 5: BA2

[0466]

[0467] To a solution of 39 (1.0 g, 1.46 mmol) in THF (1.5 mL), MeOH (1.2 mL) and H2O (0.3 mL) was added aqueous lithium hydroxide solution (5 M, 0.53 mL). After stirring at 0-25 °C for 2 h, the mixture was neutralized to ~pH 7 with citric acid (1 M) and then the volatiles were removed. The solids in the aqueous solution were then collected by filtration to afford BA2 as an off-white solid (300 mg, 54% yield). (MS: [M+H] + 372.2)

[0468]

[0469] Step 1: Trifluoroethyl ether 40

[0470]

[0471] To a solution of 22 (0.4 g, 0.93 mmol) in DMF (8 mL) was added sodium hydride (48 mg, 1.12 mmol) at 0°C, and the mixture was stirred for 30 minutes, and then 2,2,2-trifluoroethyl trifluoromethanesulfonate (0.165 mL, 1.12 mmol) was slowly added. After stirring at 0°C for 3 hours, water (3 mL) was added, the mixture was concentrated, and purified by silica gel column chromatography (EA / hexane=1 / 10) to give 40 as a white solid (218 mg, 46% yield).

[0472] Step 2: Benzoate 41

[0473]

[0474] To a solution of 40 (1.4 g, 2.74 mmol) in THF (25 mL) was added TBAF (3.3 mL, 3.3 mmol) at 0 ° C. After stirring at room temperature for 4 hours, water (2 mL) was added and the mixture was extracted with EA. The organic layer was washed with water and brine and concentrated. The residue was then dissolved in DCM, followed by the addition of TEA (1.92 mL, 13.8 mmol) and benzoyl chloride (0.42 mL, 3.6 mmol). After stirring for 1 hour, water (1 mL) was added, the mixture was concentrated, and 41 was purified by silica gel column chromatography (EA / PE=1 / 5) as a white solid (0.795 g, 77% yield).

[0475] Step 3: Acetate 42

[0476]

[0477] A solution of 41 (0.79 g, 2.1 mmol) in HOAc (17.5 mL) and water (7.5 mL) was stirred at reflux for 30 minutes, then cooled to room temperature and co-evaporated with toluene (10 mL×4). The residue was then dissolved in Py (12 mL), followed by the addition of Ac2O (0.8 mL, 8.4 mmol). After stirring for 6 hours, the mixture was concentrated and purified by silica gel column chromatography (EA / PE=1 / 4) to give 42 as a white solid (0.82 g, 91% yield).

[0478] Step 4: Guanosine 43

[0479]

[0480] 42 (800 mg, 1.9 mmol) and N were added to 80 °C. 2 - A suspension of isobutyrylguanine (633 mg, 2.86 mmol) in dichloroethane (25 mL) was added with BSA (2.74 mL, 10.1 mmol) and stirred for 1 hour, followed by the addition of TMSOTf (1.03 mL, 5.7 mmol). After stirring at 100 ° C for 3 hours, the mixture was poured into an aqueous sodium bicarbonate solution (60 mL) and extracted with DCM (60 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (MeOH / DCM=1 / 20 to 1 / 10) to give 43 as a white solid (938 mg, 85% yield).

[0481] Step 5: BA3

[0482]

[0483] To a solution of 43 (0.5 g, 0.86 mmol) in MeOH (8 mL), THF (10 mL) and water (2 mL) was added aqueous sodium hydroxide solution (10 M, 0.34 mL) at 0° C. After stirring for 30 min, HOAc was added, the mixture was concentrated, and purified by silica gel column chromatography (MeOH / DCM=1 / 10 to 1 / 5) to give BA3 as an oil (348 mg, 93% yield).

[0484]

[0485] Step 1: Pyrazole 44

[0486]

[0487] To a solution of 2 (75 g, 395 mmol) in MeOH (600 mL) was added hydrazine hydrate (120 mL) at 25 °C. After stirring for 2 hours, the mixture was concentrated, and the residue was dissolved in EtOH (600 mL), followed by the addition of (ethoxymethylene)malononitrile (110 g, 901 mmol). After stirring at 78 °C for 30 minutes, the mixture was concentrated and purified by silica gel column chromatography (EA / PE=1 / 20 to 1 / 15) to give 44 as a light yellow solid (42 g, 38% yield).

[0488] Step 2: Thiourea 45

[0489]

[0490] To a solution of 44 (16.0 g, 57 mmol) in MeOH (75 mL) and water (25 mL) was added ammonium hydroxide (280 mL) and hydrogen peroxide (150 mL). After stirring at 25 ° C for 16 hours, the mixture was poured into an aqueous sodium sulfite solution (2 L), and then extracted with EA (700 mL × 3). The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was then dissolved in acetone (90 mL), and benzoyl isothiocyanate (6.96 g, 42.7 mmol, 5.75 mL) was then added at 25 ° C. After stirring at 60 ° C for 4 hours, the mixture was concentrated to give crude compound 45 as a yellow solid.

[0491] Step 3: Pyrazolopyrimidinone 46

[0492]

[0493] To a solution of the crude compound 45 obtained above in MeOH (150 mL) was added aqueous sodium hydroxide solution (0.7 M, 80 mL), followed by iodomethane (6.8 g, 47.9 mmol, 3.0 mL). After stirring at 20 ° C for 2 hours, the mixture was neutralized to pH 6 with HOAc, then water (80 mL) was added, and extracted with EA (100 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was then dissolved in MeOH (30 mL), and aqueous sodium hydroxide solution (1.4 M, 250 mL) was added. After stirring at 100 ° C for 2 hours, the mixture was concentrated, and the residue was co-evaporated with toluene (200 mL×3) and dissolved in DCM (500 mL). Imid (18.5 g, 271 mmol), DMAP (1.66 g, 13.6 mmol) and TBSCl (40.9 g, 271 mmol) were then added. After stirring at 25°C for 18 hours, saturated aqueous sodium bicarbonate solution (1 L) was added, and the mixture was extracted with EA (500 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (MeOH / DCM = 1 / 60 to 1 / 30) to give 46 as a white solid (8.50 g, 34% yield for five steps).

[0494] Step 4: Isobutyrate 47

[0495]

[0496] To a solution of 46 (23.4 g, 53.5 mmol) in Py (120 mL) was added isobutyryl chloride (11.4 g, 107 mmol, 11.2 mL) at 25 °C. After stirring at 25 °C for 16 hours, ammonium hydroxide (0.5 mL) was added, and the mixture was stirred for 30 minutes and then concentrated. The residue was then dissolved in EtOAc (1.5 L), washed with saturated aqueous ammonium hydroxide solution (500 mL×3) and brine (500 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 10 to 1 / 3) to give 47 as a light yellow solid (24.0 g, 88% yield). (MS: [M+Na] + 530.1)

[0497] Step 5: BB1

[0498]

[0499] A solution of 47 (10.0 g, 19.7 mmol) in HOAc (6 mL) and water (3 mL) was stirred at 65 °C for 5 h. The reaction mixture was then concentrated and triturated with DCM (15 mL). The solid was collected by filtration to give crude compound BB1 as a white solid (4.0 g). (MS: [M+H] + 354.0)

[0500]

[0501] Step 1: Ketal 49

[0502]

[0503] To a solution of 48 (50 g, 135 mmol) in acetone (500 mL) was added 2,2-dimethoxypropane (85 g, 816 mmol, 100 mL) and concentrated sulfuric acid (1.32 g, 13.5 mmol, 0.72 mL). After stirring at 25 ° C for 30 minutes, saturated aqueous sodium bicarbonate solution (30 mL) was added. The solution was filtered, concentrated, and purified by silica gel column chromatography (MeOH / DCM=1 / 200 to 1 / 50) to give 49 as a white solid (35 g, 63% yield). (MS: [M+H] + 412.1)

[0504] Step 2: Azide 50

[0505]

[0506] To a solution of 49 (5.0 g, 12.2 mmol) in Py (50 mL) was added methanesulfonyl chloride (2.1 g, 18 mmol, 1.4 mL) at 0 °C. After stirring at 25 °C for 1 hour, DCM (200 mL) was added, and the solution was washed with saturated aqueous sodium bicarbonate solution, then washed with brine, dried over anhydrous sodium sulfate, and concentrated. The residue was then dissolved in DMF (50 mL), and sodium azide (3.4 g, 52.3 mmol) was added. After stirring at 50 °C for 16 hours, DCM (400 mL) was added. The mixture was washed with water (300 mL), brine, dried over sodium sulfate, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 1) to give 50 as a light yellow solid (4.0 g). (MS: [M+H] + 437.1)

[0507] Step 3: BC1

[0508]

[0509] A solution of 50 (50 g, 115 mmol) in TFA (125 mL) and water (125 mL) was stirred at 25 °C for 5 h, then concentrated, co-evaporated twice with toluene, and dissolved in MeOH (50 mL). The mixture was then neutralized with aqueous sodium bicarbonate (1%) and triturated with MTBE. The solid was collected, washed with MTBE, dried, and dissolved in DMF (400 mL). Pd / C (10% w / w, 10 g) was then added, and the mixture was stirred at 25 °C under a hydrogen atmosphere (15 psi) for 6 h, then filtered and concentrated to give crude compound BC1 as a yellow oil (39 g). (MS: [M+H] + 371.1)

[0510]

[0511] Step 1: Indole 52

[0512]

[0513] To a solution of indole (305 mg, 2.6 mmol) in MeCN (10 mL) was added sodium hydride (160 mg, 4.0 mmol) at 0 ° C and stirred for 30 minutes, then 51 (1.0 g, 2.6 mmol) was added. After stirring for 1 hour, saturated sodium bicarbonate aqueous solution (5 mL) was added, and the mixture was extracted with EA (20 mL × 3). The combined organic layer was washed with saturated sodium bicarbonate aqueous solution and brine, concentrated, and purified by silica gel column chromatography (EA / hexane=1 / 4) to give 52 as a yellow oil (886 mg, 71% yield). (MS: [M+H] + 470.2)

[0514] Step 2: BC2

[0515]

[0516] To a solution of 52 (610 mg, 1.3 mmol) in MeOH (9 mL) was added sodium methoxide (5.4 M in methanol, 0.54 mL). After stirring for 1 hour, hydrochloric acid (5 M, 0.5 mL) was added at 0 ° C, and the solution was stirred for 10 minutes, then concentrated and purified by silica gel column chromatography (MeOH / DCM=1 / 9) to give the desired product as a white solid (197 mg, 92% yield). (MS: [M+H] + 234.1)

[0517]

[0518] Step 1: Pyrene 54

[0519]

[0520] To a solution of 53 (560 mg, 2 mmol) in THF, magnesium (54 mg, 2.3 mmol) was added, followed by a small amount of iodine. After stirring at 55 ° C for 3 hours, copper (I) iodide (213 mg, 1.1 mmol) was added at 0 ° C. The mixture was stirred at room temperature for 45 minutes, and then 51 (367 mg, 0.98 mmol) was added at 40 ° C. After stirring for 2 hours, saturated aqueous ammonium chloride solution (2 mL) and DCM (20 mL) were added. Layering, and the organic layer was washed with saturated sodium bicarbonate aqueous solution and brine, dried over sodium sulfate anhydride, concentrated, and purified by silica gel column chromatography (EA / hexane=1 / 10 to 1 / 5) to obtain 54 as a white solid (52 mg, 10%).

[0521] Step 2: BC3

[0522]

[0523] To a solution of 54 (230 mg, 0.4 mmol) in MeOH (5 mL) was added sodium methoxide (30% in MeOH, 0.23 mL, 1.2 mmol) at room temperature. After stirring for 1 hour, saturated ammonium chloride (5 mL) was added, and the mixture was extracted with EA (10 mL x 3). The organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated to give crude compound BC3 as a white solid (150 mg).

[0524]

[0525] Step 1: Alcohol 56

[0526]

[0527] To a solution of 55 (91.4 g, 481 mmol) in Py (600 mL) was added trityl chloride (160.7 g, 577 mmol). After stirring at 60 °C for 16 hours, the mixture was concentrated and co-evaporated with toluene three times. The residue was partitioned between DCM (400 mL) and saturated aqueous sodium bicarbonate solution (750 mL). The layers were separated and the aqueous phase was extracted with DCM (400 mL×2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 10 to 1 / 5) to give 56 as a white solid (180.5 g, 87% yield). (MS: [M+Na] + 455.0)

[0528] Step 2: Keto 57

[0529]

[0530] To a solution of 56 (176 g, 407 mmol) in MeCN (1.0 L) was added IBX (228 g, 814 mmol). After stirring at 90 °C for 6 h, the mixture was filtered and concentrated to give crude compound 57 as a light yellow oil (175 g). (MS: [M+Na] + 453.0)

[0531] Step 3: Ester 58

[0532]

[0533] To a solution of sodium hydride (20.1 g, 502 mmol) in THF (1.0 L) was added dropwise 2-dimethoxyphosphoryl acetate (96.3 g, 529 mmol, 76.5 mL) over 15 minutes at 0°C. After stirring for 60 minutes, the crude compound 57 (175 g) obtained above in THF (500 mL) was added dropwise at 0°C. After stirring for 16 hours at 25°C, water (50 mL) was added at 0°C, and volatiles were removed, and brine (500 mL) was added. The mixture was then extracted with DCM (500 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue (198 g of the 216 g obtained above) was then dissolved in EA (500 mL), and Pd / C (10% w / w, 10 g) was added. After stirring at 25°C under hydrogen atmosphere (20 psi) for 16 hours, the mixture was filtered, and the filtrate was concentrated and purified by silica gel column chromatography (EA / PE = 1 / 15 to 1 / 10) to give 58 as a white solid (120 g, 66% yield). (MS: [M+Na] + 511.1)

[0534] Step 4: Alcohol 59

[0535]

[0536] To a solution of lithium aluminum hydride (6.21 g, 164 mmol) in THF (200 mL) was slowly added 58 (20.0 g, 40.9 mmol) in THF (50 mL) at 0°C. After stirring at 25°C for 16 hours, the reaction was quenched by sequentially adding water (6.2 mL), aqueous sodium hydroxide solution (15%, 6.2 mL) and water (18.6 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 10 to 1 / 4) to give 59 as a white solid. (MS: [M+Na] + 483.2)

[0537] Step 5: Alcohol 60

[0538]

[0539] To a solution of sodium hydride (60% w / w, 6.95 g, 174 mmol) in THF (200 mL) was added 59 (20.0 g, 43.4 mmol) in THF (80 mL) at -20 °C over 5 minutes. After stirring at 25 °C for 2 hours, benzyl bromide (22.3 g, 130 mmol, 15.5 mL) was added dropwise, and the mixture was stirred at 80 °C for 16 hours, and then water (2 mL) was added at 0 °C. The mixture was diluted with water (200 mL) and extracted with DCM (200 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated. The residue was then dissolved in DCM (200 mL), and DCA (5.48 g, 42.5 mmol, 12.0 mL) was added. After stirring at 25 °C for 3 hours, saturated aqueous sodium bicarbonate solution was added at 0 °C. The mixture was then extracted with DCM (150 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE = 1 / 5 to 1 / 2) to give 60 as a yellow oil (12.1 g, 90% yield).

[0540] Step 6: Benzoate 61

[0541]

[0542] To a solution of 60 (24.0 g, 78 mmol) in DCM (500 mL) was added benzoyl chloride (16.4 g, 116.7 mmol, 13.6 mL) and TEA (23.6 g, 233.5 mmol, 32.4 mL). After stirring at 25 °C for 16 hours, the mixture was concentrated and purified by silica gel column chromatography (EA / PE=1 / 15 to 1 / 10) to give 61 as a light yellow oil (30.0 g, 93% yield). (MS: [M+Na] + 435.1)

[0543] Step 7: Acetate 62

[0544]

[0545] A mixture of 61 (29.0 g, 70.3 mmol) and water (3.0 mL) in HOAc (220 mL) was stirred at 70 ° C for 16 hours, and then a saturated aqueous sodium bicarbonate solution was added. The mixture was then extracted with DCM (400 mL × 3). The combined organic layers were concentrated, and the residue was dissolved in Py (30 mL), followed by the addition of Ac2O (28.5 g, 280 mmol, 26 mL). After stirring at 20 ° C for 16 hours, a saturated aqueous sodium bicarbonate solution was added, and the mixture was then extracted with DCM (500 mL × 3). The combined organic layers were concentrated, and 62 was purified by silica gel column chromatography (EA / PE = 1 / 10 to 1 / 5) to give a white solid (31.1 g, 97% yield). (MS: [M + Na] + 479.1)

[0546] Step 8: Purine 63

[0547]

[0548] At 20℃, 6 -Diphenylcarbamoyl-N 2 -BSA (11.6 g, 57.0 mmol, 14.1 mL) was added to a suspension of isobutyrylguanine (5.47 g, 13.1 mmol) in MeCN (150 mL). After stirring at 63 °C for 30 min, the volatiles were removed and the residue was dissolved in MeCN (200 mL), and then 62 (5.00 g, 11.0 mmol) in MeCN (50 mL) and TMSOTf (3.65 g, 16.4 mmol, 3.0 mL) was added at -15 °C. After stirring at 63 °C for 50 min, the mixture was cooled to 0 °C, poured into a saturated aqueous sodium bicarbonate solution, and extracted with EA (150 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, concentrated, and purified by silica gel flash chromatography (EA / PE=1 / 3 to 1 / 1) to give 63 as a white solid. (MS: [M+H] + 813.1)

[0549] Step 9: Guanosine 64

[0550]

[0551] A solution of 63 (16.2 g, 19.9 mmol) in 90% TFA aqueous solution (60 mL) was stirred at 20°C for 30 minutes, then poured into saturated sodium bicarbonate aqueous solution at 0°C and extracted with EA (100 mL×4). The combined organic layers were dried over anhydrous sodium sulfate, concentrated, and purified by silica gel flash chromatography (EA / PE=1 / 1 to 1 / 0) to give 64 as a white solid (11.4 g, 93% yield). (MS: [M+H] + 618.1)

[0552] Step 10: Silyl ether 65

[0553]

[0554] To a solution of 64 (15.0 g, 24.3 mmol) in EtOH (500 mL) was added Pd / C (10% w / w, 2.0 g) and concentrated hydrochloric acid (10 drops). After stirring at 50°C under a hydrogen atmosphere (45 psi) for 15 hours, the mixture was filtered and the solid was washed with EtOH (100 mL×3). The filtrate was concentrated and one-third of the residue was dissolved in DMF (60 mL), followed by the addition of Imid (1.57 g, 23.0 mmol), DMAP (46.9 mg, 0.38 mmol) and triisopropylsilyl chloride (2.22 g, 11.5 mmol, 2.5 mL). After stirring at 20°C for 16 hours, saturated aqueous sodium bicarbonate solution (20 mL) and water (100 mL) were added. The mixture was then extracted with EA (100 mL×2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 5 to 2 / 1) to give 65 as a white solid (4.52 g, 86% yield). (MS: [M+H] + 684.4)

[0555] Step 11: BC4

[0556]

[0557] To a solution of 65 (3.0 g, 4.4 mmol) in EtOH (30 mL) was added aqueous sodium hydroxide solution (2 M, 31 mL) at 0°C. After stirring at 0°C for 30 min, the mixture was neutralized by adding hydrochloric acid solution (1 N) and HOAc at 0°C. Toluene (30 mL) was then added, and the mixture was concentrated to give crude compound BC4 as a white solid (3.0 g). (MS: [M+H] + 538.2)

[0558]

[0559] Step 1: Alcohol 66

[0560]

[0561] 1,3-Dichloro-1,1,3,3-tetraisopropyldisiloxane (1.79 g, 5.66 mmol, 1.8 mL) was slowly added to a solution of 15 (2.0 g, 5.66 mmol) in Py (56 mL) at 0°C. After stirring at 0°C for 30 minutes and at 25°C for 12 hours, the solution was concentrated and purified by silica gel column chromatography (MeOH / DCM=1 / 20) to give 66 (1.6 g, 47% yield). (MS: [M+H] + 596.3)

[0562] Step 2: Purine 67

[0563]

[0564] To a solution of 66 (8.0 g, 13.5 mmol) and 2-(4-nitrophenyl)ethanol (3.37 g, 20.2 mmol) in THF (100 mL) was slowly added DIAD (6.81 g, 33.7 mmol, 6.6 mL) and PPh3 (8.83 g, 33.7 mmol) at 25°C. After stirring at 25°C for 12 hours, water (5 mL) was added, the mixture was concentrated, and purified by silica gel column chromatography (MeOH / DCM=1 / 20) to give 67 as a light yellow solid (4.5 g, 44% yield). (MS: [M+H] + 745.3)

[0565] Step 3: Alcohol 68

[0566]

[0567] To a solution of 67 (4.20 g, 5.64 mmol) in MeCN (40 mL) at 25 °C was added IBX (3.16 g, 11.3 mmol). After stirring at 80 °C for 12 hours, the mixture was filtered and concentrated, and dissolved in THF (50 mL). Sodium triacetoxyborohydride (5.7 g, 27.0 mmol) was then slowly added at 0 °C. After stirring at 25 °C for 6 hours, water (5 mL) was added, and the mixture was concentrated and purified by silica gel column chromatography (MeOH / DCM=1 / 20) to give 68 as a light yellow oil (1.0 g, 33% yield). (MS: [M+H] + 745.3)

[0568] Step 4: Azide 69

[0569]

[0570] To a solution of 68 (2.2 g, 2.95 mmol) and DMAP (1.44 g, 11.8 mmol) in DCM (140 mL) and Py (10 mL) was slowly added trifluoromethanesulfonic anhydride (1.33 g, 4.72 mmol, 0.78 mL) at 0 °C. After stirring at 0 °C for 1.5 h, the mixture was concentrated. The residue was then dissolved in DMF (10 mL) and sodium azide (0.49 g, 7.53 mmol) was added. After stirring at 60 °C for 6 h, the solution was concentrated and purified by preparative HPLC (MeOH / water containing 0.1% HCOOH: 40–100%) to give 69 as a light yellow solid (1.50 g, 79% yield). (MS: [M+H] + 770.4)

[0571] Step 5: BC5

[0572]

[0573] To a solution of 69 (2.50 g, 3.25 mmol) in THF (12 mL) was slowly added TBAF (13.1 g, 50.1 mmol) and HOAc (1.50 g, 25.0 mmol, 1.43 mL) at 15 °C. After stirring at 15 °C for 12 hours, the mixture was concentrated. The residue was then dissolved in DCM (20 mL), washed with water (5 mL x 2), dried over anhydrous sodium sulfate, filtered, and purified by silica gel column chromatography (MeOH / DCM = 1 / 20) to give BC5 as a light yellow solid (900 mg, 53% yield). (MS: [M+H] + 528.2)

[0574] The following compounds were prepared essentially by the methods described above for intermediates BC3 and BC4.

[0575] Table 1. Intermediates BC6 and BC7

[0576]

[0577]

[0578] Step 1: Paraben 70

[0579]

[0580] To a solution of 33 (120 g, 449 mmol) in Py (1.0 L) was added TMSCl (390 g, 3.59 mol, 454 mL). After stirring at 0°C for 2 hours, benzoyl chloride (316 g, 2.25 mol, 261 mL) was added dropwise, and the mixture was stirred at 25°C for 14 hours, then cooled to 0°C. Water (240 mL) was then added, and the mixture was stirred at 25°C for 30 minutes, then ammonium hydroxide (460 mL) was added at 0°C. After stirring for 2 hours, the mixture was concentrated to give 70 as a white solid (150 g, 90% yield).

[0581] Step 2: Diol 71

[0582]

[0583] To a solution of 70 (150 g, 404 mmol) in Py (500 mL) was added DMTrCl (274 g, 808 mmol), TEA (81.8 g, 808 mmol, 112 mL) and DMAP (4.93 g, 40.4 mmol) at 0°C. After stirring at 25°C for 16 hours, saturated aqueous sodium bicarbonate solution (1 L) was added, and the mixture was extracted with EtOAc (600 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE 1 / 4 to 1 / 2, then MeOH / DCM 1 / 100 to 1 / 20) to give 71 as a white solid (65.0 g, 24% yield).

[0584] Step 3: Silyl ether 72

[0585]

[0586] To a solution of 71 (65.0 g, 96.5 mmol) in Py (500 mL) was added silver nitrate (32.8 g, 193 mmol) and TBSCl (29.1 g, 193 mmol) at 25°C. After stirring at 25°C for 1 hour, saturated aqueous sodium bicarbonate solution (1 L) was added, and the mixture was extracted with EtOAc (600 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE = 1 / 4 to 1 / 1) to give 72 as a white solid (20.0 g, 26% yield).

[0587] Step 4: A2

[0588]

[0589] To a solution of 72 (12.0 g, 15.2 mmol) in DIPEA (15 mL) and DCM (30 mL) were added DMAP (744 mg, 6.09 mmol) and 2-cyanoethyl N,N-diisopropylchlorophosphoramidite (5.41 g, 22.9 mmol) at 25° C. After stirring for 2 hours, the mixture was directly purified by basic silica gel column chromatography (EA / PE=1 / 4 to 1 / 1) to give A2 as a white solid (13.0 g, 86% yield).

[0590] Step 5: A1

[0591]

[0592] To a solution of A2 (3.8 g, 3.9 mmol) in MeCN (20 mL) was added water (0.1 mL) and pyridinium trifluoroacetate (1.1 g, 5.8 mmol) at 25 ° C and stirred for 5 minutes, followed by the addition of tert-butylamine (14.0 g, 0.19 mmol). After stirring for 15 minutes, the volatiles were removed and the residue was dissolved in DCM (20 mL). A solution of DCA (1.9 g, 14.6 mmol) in DCM (20 mL) was then added. After stirring for 30 minutes, TEA (3 mL) was added, the mixture was concentrated, and the A1 · TEA salt was purified by reverse phase silica gel column chromatography (MeCN / water containing 0.1% TEA = 0% to 100%) as a white solid (1.5 g, 71% yield). (MS: [M+H] + 549.2)

[0593] Preparation of AB1:

[0594]

[0595] To a solution of A2 (494 mg, 0.5 mmol) and triphenylphosphine (197 mg, 0.75 mmol) and 2-(tert-butyldimethylsilyloxy)ethanol (132 mg, 0.75 mmol) in THF (5 mL) was added DIAD (0.15 mL, 0.75 mmol). After stirring at room temperature for 5 hours, the mixture was concentrated and purified by silica gel column chromatography (EA / hexane=1 / 9 to 1 / 4) to give AB1 as a white solid (230 mg, 40% yield).

[0596]

[0597] Step 1: Diol 73

[0598]

[0599] To a solution of crude BCl (39 g) in Py (40 mL) was added DMTrCl (35.9 g, 106 mmol) at 0 °C. After stirring at 25 °C for 16 h, MeOH (50 mL) was added and the mixture was concentrated. The residue was then dissolved in DCM (600 mL), washed with saturated aqueous sodium bicarbonate solution and brine, concentrated, and purified by silica gel column chromatography (MeOH / DCM=1 / 100 to 1 / 50) to give 73 as a light yellow solid (34.0 g, 48% yield for two steps). (MS: [M+H] + 673.2)

[0600] Step 2: Silyl ether 74

[0601]

[0602] To a solution of 73 (1.0 g, 1.49 mmol) in Py (10 mL) was added silver nitrate (380 mg, 2.24 mmol, 0.38 mL) at 0°C. After stirring for 15 minutes, TBSCl (270 mg, 1.79 mmol) was added, and the mixture was stirred at 25°C for 2 hours, and then saturated aqueous sodium bicarbonate solution was added. The mixture was then extracted with EA (10 mL), the organic layer was washed with brine, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 5) to give 74 as a light yellow solid (400 mg, 34% yield). (MS: [M+H] + 787.3)

[0603] Step 3: AC1

[0604]

[0605] To a solution of 74 (1.0 g, 1.27 mmol) in Py (10 mL) was added diphenyl phosphite (80%, 744 mg, 2.54 mmol, 0.61 mL). After stirring at 20 °C for 1 hour, EA (2 mL) and saturated aqueous sodium bicarbonate solution (2 mL) were added, and the mixture was stirred for 1 hour. The layers were separated and the organic layer was concentrated. The residue was then dissolved in DCM (1.0 mL), and DCA (164 mg, 1.27 mmol, 0.1 mL) was added. After stirring at 25 °C for 30 minutes, TEA (1 mL) was added, the filtrate was concentrated, and AC1 was purified by reverse phase silica gel column chromatography (MeCN / water containing 0.1% TEA = 0% to 100%) to give AC1 as a white solid (500 mg, 72% yield). (MS: [M+H] + 549.2)

[0606]

[0607] Step 1: Alcohol 75

[0608]

[0609] To a solution of BC2 (348 mg, 1.49 mmol) in Py (15 mL) was added DMAP (18 mg, 0.15 mmol) and DMTrCl (0.66 g, 1.94 mmol). After stirring overnight, MeOH (3 mL) was added, the mixture was concentrated, and purified by silica gel column chromatography (EA / hexane=1 / 4) to give 75 (662 mg, 83% yield). (MS: [M+H] + 536.2)

[0610] Step 2: AC2

[0611]

[0612] To a solution of 75 (0.2 g, 0.37 mmol) in DCM (4 mL) was added DIEPA (0.15 g, 1.2 mmol, 0.2 mL) and 2-cyanoethyl N,N-diisopropylchlorophosphoramidite (0.14 g, 0.56 mmol, 0.13 mL). After stirring for 4 hours, the mixture was concentrated and purified by silica gel column chromatography (EA / hexane containing 1% TEA = 1 / 4) to give AC2 (232 mg, 85% yield). (MS: [M–N i Pr2+H2O] + 653.2)

[0613]

[0614] Step 1: Silyl ether 76

[0615]

[0616] To a solution of 16 (11.0 g, 16.8 mmol) in DCM (80 mL) was added TBSCl (7.59 g, 50.3 mmol) and Imid (3.43 g, 50.3 mmol). After stirring at 25 °C for 16 hours, an aqueous sodium bicarbonate solution (5%, 30 mL) was added, and the mixture was extracted with DCM (60 mL×3). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by basic silica gel column chromatography (EA / PE=1 / 5 to 1 / 1) to give 76 as a white solid (2.1 g, 16% yield).

[0617] Step 2: G2

[0618]

[0619] To a solution of 76 (900 mg, 1.17 mmol) in THF (4.0 mL) and DIEPA (4.0 mL) was added DMAP (14.3 mg, 0.12 mmol) dropwise at 0°C, followed by dropwise addition of 2-cyanoethyl N,N-diisopropylchlorophosphoramidite (415 mg, 1.76 mmol). After stirring at 20-25°C for 2 hours, an aqueous sodium bicarbonate solution (5%, 15 mL) was added at 0°C. The mixture was then diluted with water (15 mL) and extracted with EA (15 mL×3). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (acetone / PE=1 / 10 to 1 / 3) to give G2 as a white solid (600 mg, 53% yield).

[0620] Step 3: G1

[0621]

[0622] To a solution of G1 (7.0 g, 7.22 mmol) in MeCN (30 mL) was added water (0.11 mL) and pyridinium trifluoroacetate (4.18 g, 21.7 mmol) at 25 ° C. After stirring at 25 ° C for 15 minutes, tert-butylamine (37 mL) was added, and the mixture was stirred at 25 ° C for 45 minutes and then concentrated. The residue was then dissolved in DCM (30 mL), and a solution of DCA in DCM (6% v / v, 30 mL) was added dropwise. After stirring at 20-25 ° C for 30 minutes, DCM (30 mL) and TEA (4 mL) were added. The mixture was then concentrated, dissolved in a mixture of MeCN (5 mL) and water (5 mL), and purified by C18 reverse phase medium pressure liquid chromatography (MeCN / water containing 0.1% TEA = 0% to 60%) to give G1.TEA salt as a yellow solid (2.30 g, 56% yield). (MS:[M+H] + 532.3)

[0623]

[0624] Step 1: Diol 77

[0625]

[0626] To two batches of crude BB1 ​​(8.0 g) obtained above in Py (50 mL) was added DMTrCl (9.2 g, 27.2 mmol). After stirring at 20-30°C for 1 hour, MeOH (10 mL) was added, and the mixture was concentrated and purified by silica gel column chromatography (EA / PE=1 / 5 to MeOH / DCM=1 / 20) to give 77 as a yellow solid (11.0 g, 30% yield for two steps). (MS: [M+Na] + 678.2)

[0627] Step 2: Silyl ether 78

[0628]

[0629] To a solution of 77 (9.0 g, 14.0 mmol) in Py (50 mL) was added TBSCl (2.48 g, 16.5 mmol) and silver nitrate (5.83 g, 34.3 mmol). After stirring at 25-30 ° C for 30 minutes, a saturated aqueous sodium bicarbonate solution was added. The mixture was then extracted with DCM (200 mL×2), and the combined organic layers were washed with water (50 mL), brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 10 to 1 / 5 to 1 / 3) to give 78 as a white foam (1.50 g, 14% yield). (MS: [M+Na] + 792.2)

[0630] Step 3: GB1

[0631]

[0632] To a solution of 78 (2.50 g, 3.3 mmol) in DIEPA (5 mL) and DCM (5 mL) was added DMAP (200 mg, 1.62 mmol) and 2-cyanoethyl N,N-diisopropylchlorophosphoramidite (1.0 g, 4.22 mmol). After stirring at 20-25 °C for 2 h, the mixture was concentrated and purified by silica gel column chromatography (EA / PE = 1 / 10 to 1 / 4) to give GB1 as a white foam (2.30 g, 73% yield).

[0633] Step 4: GB2

[0634]

[0635] To a solution of GB1 (2.30 g, 2.4 mmol) in MeCN (2.0 mL) was added water (0.11 mL, 6.1 mmol) and pyridinium trifluoroacetate (687 mg, 3.56 mmol) at 25°C. After stirring at 25-30°C for 30 minutes, the mixture was concentrated and the residue was dissolved in MeCN (20 mL), followed by the addition of tert-butylamine (10.5 g, 144 mmol, 15.0 mL). After stirring at 25-30°C for 30 minutes, the mixture was concentrated and DCM (20 mL) was added, followed by a solution of DCA in DCM (6% v / v, 18.2 mL). The mixture was stirred at 25-30 °C for 30 min, then neutralized with TEA to ~pH 7, concentrated, and purified by C18 reverse phase silica gel column chromatography (MeCN / water containing 0.1% TEA = 0% to 40%) to give GB2 as a white solid (800 mg, 63% yield). (MS: [M+H] + 532.0)

[0636]

[0637] Step 1: Alcohol 79

[0638]

[0639] To a solution of BC4 (1.97 g, 2.4 mmol) in Py (20 mL) was added DMTrCl (984 mg, 2.90 mmol) at 25 °C. After stirring for 3 h, MeOH (30 mL) was added, the mixture was concentrated, and purified by basic silica gel flash chromatography (EA / PE=1 / 5 to 4 / 1) to give 79 as a light yellow powder (1.65 g, 82% yield). (MS: [M+H] + 840.2)

[0640] Step 2: GC1

[0641]

[0642] To a solution of 79 (2.0 g, 2.38 mmol) in Py (15 mL) was added diphenyl phosphite (1.7 g, 7.1 mmol, 1.4 mL). After stirring at 20 ° C for 30 minutes, saturated sodium bicarbonate aqueous solution (30 mL) was added, and the mixture was stirred for 1 hour. The mixture was then extracted with EA (30 mL × 3). The combined organic layer was washed with saturated sodium bicarbonate aqueous solution (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was then dissolved in DCM (40 mL), followed by the addition of water (0.4 mL) and DCA (6% v / v in DCM, 40 mL). After stirring at 20 ° C for 15 minutes, the mixture was neutralized with TEA to ~ pH 7, concentrated, and purified by reverse phase silica gel column chromatography (MeCN / water = 25% to 90%) to give GC1 as a white solid (1.3 g, 90% yield). (MS: [M + H] + 602.1)

[0643]

[0644] Step 1: Alcohol 80

[0645]

[0646] To a solution of BC5 (900 mg, 1.71 mmol) in Py (10 mL) was added DMTrCl (809 mg, 2.39 mmol) at 15°C. After stirring at 15°C for 12 h, MeOH (0.5 mL) was added, the mixture was concentrated, and purified by silica gel column chromatography (MeOH / DCM=1 / 50) to give 80 as a yellow oil (1.0 g, 71% yield). (MS: [M+H] + 528.2)

[0647] Step 2: GC2

[0648]

[0649] To a solution of 80 (2.05 g, 2.47 mmol) in DMF (5.0 mL) at 15 ° C was added Imid (673 mg, 9.88 mmol) and TBSCl (745 mg, 4.94 mmol, 0.61 mL). After stirring at 15 ° C for 12 hours, the mixture was concentrated and the residue was ground with water (10 mL). The solid was then collected and washed with water (10 mL×2), PE (10 mL×2), dried, and dissolved in THF (18 mL), and then PPh3 (1.11 g, 4.24 mmol) was added at 15 ° C. After stirring at 15 ° C for 2.5 hours, water (0.16 mL) was added and the mixture was stirred at 50 ° C for 12 hours. The solution was then concentrated and purified by reverse phase preparative HPLC (MeOH / water containing 0.1% TEA = 20% to 80%) to give GC2 as a white solid (900 mg, 58% yield). (MS: [M+H] + 918.1)

[0650] The following compounds were prepared essentially by the methods described above for Intermediates A1, A2, AC1, AC2, G1, G2, GC1 and GC2.

[0651]

[0652]

[0653]

[0654]

[0655]

[0656]

[0657]

[0658]

[0659]

[0660]

[0661]

[0662]

[0663]

[0664]

[0665]

[0666]

[0667] To a solution of crude G1 (obtained from 187 mg of G2·TEA salt, 0.2 mmol, containing Py·DCA salt) in MeCN (0.5 mL) was added a solution of A2 (0.26 g, 0.26 mmol) in MeCN (0.2 mL). After stirring for 30 minutes, DDTT (46 mg, 0.22 mmol) was added, and the mixture was stirred for 1 hour and then concentrated. The residue was dissolved in DCM (4.8 mL), and water (0.036 mL) and DCA (6% DCM solution, 4.8 mL) were added. After stirring for 10 minutes, Py (1 mL) was added, the mixture was concentrated, and the C1·Py salt was purified by silica gel column chromatography (MeOH / DCM=1 / 10 to 1 / 5) as a white solid (86 mg, 35% yield). (MS: [M+H] + 1148.0)

[0668]

[0669] To a mixture of A1 (1.0 g, 1.82 mmol, co-evaporated MeCN 20 mL×3) and G2 (2.3 g, 2.37 mmol, co-evaporated with MeCN 20 mL×3) was added tetrazole (0.45 M in MeCN, 10 mL) at 25°C and stirred for 1 hour, then elemental sulfur (1.75 g, 6.84 mmol) was added. After stirring for 1 hour, MeCN (20 mL) was added, and the mixture was filtered and concentrated. The residue was dissolved in DCM (100 mL), and DCA (1.96 g, 15.2 mmol, 1.25 mL) was added. After stirring for 2 hours at 25°C, saturated aqueous sodium bicarbonate solution (100 mL) was added. The layers were separated, and the aqueous layer was extracted with EA (100 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse phase silica gel column chromatography (MeCN / water containing 0.1% TEA = 0% to 100%) to give C2.TEA salt as a white solid (100 mg, 5% yield). (MS: {[M+2H] 2+} / 2 574.6)

[0670]

[0671] A2 (510 mg, 0.52 mmol, co-evaporated with MeCN 5 mL×3) was added with To a solution of MS (100 mg) in MeCN (1 mL) treated for 30 min was added G1 (250 mg, 0.47 mmol, co-evaporated with MeCN 5 mL×3) and pyridinium trifluoroacetate (109 mg, 0.56 mmol, co-evaporated with MeCN 5 mL×3) and then added with MS (50mg) was used to treat the mixture in MeCN (1.5mL) for 30 minutes. After stirring for 4 hours, TBHP (5.5M in decane, 0.26mL) was added, and the mixture was stirred for 30 minutes, and then sodium bisulfite aqueous solution (33%, 0.24mL) was added at 0°C. The mixture was then stirred at room temperature for 10 minutes, then filtered and concentrated. The residue was dissolved in DCM (6.2mL), followed by water (0.09mL) and DCA (0.37mL) in DCM (6.2mL). After stirring at room temperature for 10 minutes, Py (0.73mL, 9.05mmol) and DCM (35mL) were added. The mixture was washed with water (10mL×2), and the combined organic layer was extracted with dichloromethane (10mL×2). The combined organic layers were dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (MeOH / CH2Cl2 / Py=10:89.5:0.5 to 25:74.5:0.5) to give C3 as a white solid (250 mg, 47%). (MS: [M+H] + 1132.2)

[0672]

[0673] G1 (500 mg, 0.94 mmol, co-evaporated with MeCN 10 mL×3) and pyridinium trifluoroacetate (218 mg, 1.13 mmol, co-evaporated with MeCN 10 mL×3) were added with A solution of A2 (976 mg, 0.99 mmol, co-evaporated with MeCN 10 mL×3) was added to a solution of MS (100 mg) in MeCN (3 mL) for 30 min. MS (200 mg) was used to treat a solution in MeCN (2 mL) for 30 minutes. After stirring at room temperature for 2.5 hours, the mixture was concentrated and co-evaporated with MeCN (10 mL × 2). The residue was then dissolved in DCM (20 mL), followed by dropwise addition of borane dimethyl sulfide complex (2M in THF, 0.94 mL, 1.88 mmol). After stirring at room temperature for 1 hour, MeOH (0.17 mL) was added at 0 ° C, stirred for 20 minutes, and then concentrated to give crude CA1.

[0674]

[0675] Step 1: Squaramide 81

[0676]

[0677] 3,4-dimethoxy-3-cyclobutene-1,2-dione (186 mg, 1.31 mmol) was slowly added to a solution of GC2 (802 mg, 0.87 mmol) in DMF (5 mL) at 15°C. After stirring for 2 hours at 15°C, the mixture was concentrated and purified by reverse phase preparative HPLC (MeCN / water containing 0.1% TEA = 0% to 100%) to give a mixture of the desired product and unidentified by-products (0.7 g, about 84% purity). (MS: [M+H] + 1028.4)

[0678] Step 2: CC1

[0679]

[0680] To a solution of 81 (about 84% pure, 0.6 g) and AC1 (0.64 g, 1.17 mmol) obtained above in DMF (5.0 mL) was added TEA (177 mg, 1.75 mmol, 0.24 mL) at 15°C. After stirring at 15°C for 12 hours, the mixture was concentrated, and the residue was dissolved in DCM (5.0 mL), and then DCA (470 mg, 3.65 mmol, 0.3 mL) was added. The mixture was then stirred at 15°C for 15 minutes and then concentrated. The residue was purified by reverse phase preparative HPLC (MeCN / water containing 0.1% TEA = 0% to 100%) to give CC1 as a white solid (0.54 g, 40% yield for two steps). (MS: [M+H] + 1242.3)

[0681]

[0682] Step 1: Sulfamate 82

[0683]

[0684] To a solution of 4-nitrophenyl chlorosulfate (0.31 g, 1.31 mmol) in DCM (1.0 mL) was added GC2 (0.40 g, 0.44 mmol), 4-nitrophenol (0.61 g, 4.4 mmol) and TEA (0.73 mL, 5.23 mmol) in DCM (5 mL) at -78 ° C. After stirring for 30 minutes, the mixture was warmed to room temperature, diluted with DCM (20 mL), and washed with water (20 mL×3). The combined aqueous layer was extracted with DCM (20 mL×2), and the combined organic layer was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by silica gel column chromatography (EA / PE=1 / 5 to 1 / 2 containing 1% TEA) to give 82 (0.30 g, 59% yield) as a white solid. (MS: [M+H] + 1118.9)

[0685] Step 2: CC2

[0686]

[0687] 82 (0.2 g, 0.18 mmol) was dissolved in THF (1.0 mL), A solution of MS (0.05 g) and TEA (0.12 mL, 0.89 mmol) was added to AC1 (0.12 g, 0.21 mmol). After stirring for 12 h, the mixture was diluted with THF (2 mL), filtered, and concentrated. The residue was then dissolved in DCM (5.0 mL), followed by the addition of water (0.1 mL) and a solution of DCA (0.46 mL) in DCM (5.0 mL). After stirring for 15 min, the mixture was neutralized with TEA to ~pH 7, then concentrated and purified by reverse phase C18 silica gel column chromatography (MeCN / water containing 0.5% TEA = 0% to 40%) to give CC2 (0.11 g, 44% yield) as a white solid. (MS: [M+H] + 1226.0)

[0688] Example

[0689]

[0690] Step 1: C4

[0691]

[0692] To a solution of A4 (1.0 g, 1.01 mmol) and G1 (1.07 g, 2.02 mmol) in MeCN (10 mL) was added Py·DCA (420 mg, 2.02 mmol). After stirring at 20° C. for 2 hours, TBHP (70% aqueous solution, 0.65 mL, 5.05 mmol) was added, and the mixture was stirred for 1 hour, and then aqueous sodium bicarbonate solution (50 mL) was added. The mixture was extracted with ethyl acetate (100 mL), and the organic layer was washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was then dissolved in a mixture of DCM (20 mL), TFA (1.0 mL), and triethylsilane (5.0 mL). After stirring for 2 hours, the mixture was neutralized to ~pH 7 with solid sodium bicarbonate. The mixture was then filtered, and the solid was washed with EA (50 mL×3). The filtrate was concentrated and purified by preparative HPLC (MeCN / water containing 0.1% TEA = 0% to 30%) to give C4.TEA as a white solid (620 mg, 49% yield). (MS: [M+H] + 1131.1)

[0693] Step 2: Phosphodiester 83

[0694]

[0695] To a solution of C4 (583 mg, 0.515 mmol) in Py (10 mL) was added DMOCP (583 mg, 3.16 mmol) at 20 °C. After stirring for 2 hours, iodine (654 mg, 2.58 mmol) was added, and the mixture was stirred for 1 hour, followed by the addition of saturated aqueous sodium sulfate solution (30 mL) and saturated aqueous sodium bicarbonate solution (30 mL). The mixture was then extracted with EA (100 mL), washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse phase silica gel column chromatography (MeCN / water containing 0.1% TEA = 20% to 40%) to give 83.TEA as a white solid (172.0 mg, 31% yield). (MS: [M+H] + 1076.1)

[0696] Step 3: Diphosphodiester 84

[0697]

[0698] A solution of 83 (100 mg, 0.093 mmol) in MeOH (1.0 mL) and ammonium hydroxide (1.0 mL) was stirred at 50 °C for 12 h. The mixture was then concentrated and purified by reverse phase silica gel column chromatography (MeCN / water containing 0.1% TEA = 20% to 40%) to give 84 as a yellow solid (27.0 mg, 32% yield). (MS: [M+H] + 902.5)

[0699] Step 4: E5

[0700]

[0701] A solution of 84 (27 mg, 0.030 mmol) in TEA·3HF (10 mL) was stirred at 50°C for 3 h. The mixture was then neutralized with cold triethylammonium bicarbonate to ~pH 7, concentrated, and purified by C18 reverse phase silica gel column chromatography (MeCN / water containing 0.1% TEA = 0% to 20%) to give E5·TEA as a white solid (5.2 mg, 26% yield). (MS: [M+H] + 673.7)

[0702]

[0703] Step 1: C5

[0704]

[0705] To a mixture of A4 (500 mg, 0.50 mmol, co-evaporated with MeCN 5 mL×1 and toluene 10 mL×2) and G1 (322 mg, 0.61 mmol, co-evaporated with MeCN 5 mL×1 and toluene 10 mL×2) was added tetrazole (0.45 M in MeCN, 4.0 mL). After stirring at 25°C for 2 hours, DDTT (240 mg, 1.2 mmol) was added, and the mixture was stirred for 16 hours, then filtered and concentrated. The residue was then dissolved in DCM (10 mL), followed by water (0.1 mL) and DCA (0.21 mL). After stirring for 10 minutes, TEA (1 mL) was added, the mixture was concentrated, and purified by reverse phase silica gel column chromatography (MeCN / water containing 0.1% TEA = 0% to 100%) to give C5·TEA as a white solid (250 mg, 35% yield). (MS: {[M+2H] 2+ ) / 2 574.6)

[0706] Step 2: Phosphorothioate 85

[0707]

[0708] To a solution of C5 (600 mg, 0.423 mmol, co-evaporated with Py 3 mL×2) in Py (5.0 mL) was added DMOCP (313 mg, 1.69 mmol) at 25°C. After stirring for 2 hours, 3H-1,2-benzodithiol-3-one (142 mg, 0.85 mmol) was added, and the mixture was stirred for 2 hours before adding aqueous sodium bicarbonate solution (5%, 10 mL). The mixture was then extracted with EA (10 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse phase HPLC (MeCN / water containing 0.1% TEA = 0 to 100%) to give four diastereomers of 85·TEA as white solids. Isomer 1 (28 mg) (MS: [M+H] + 1160.9); Isomer 2 (25 mg) (MS: [M+H] + 1160.9); Isomer 3 (50 mg) (MS: [M+H] + 1160.9); Isomer 4 (52 mg) (MS: [M+1] + 1160.9)

[0709] Step 3: E15-E18

[0710]

[0711] Each isomer of 85.TEA (25 mg, 0.022 mmol) was stirred in ammonium hydroxide (5.6 mL) and MeOH (4.0 mL) at 50 ° C for 16 hours. The mixture was then purged with nitrogen at room temperature for 5 minutes and then concentrated. The residue was dissolved in TEA (0.5 mL) and Py (0.2 mL), and TEA.3HF (0.7 mL) was added. After stirring at 50 ° C for 24 hours, triethylammonium bicarbonate aqueous solution (1 M, 5 mL) was added, and the mixture was purified by reverse phase silica gel column chromatography (MeCN / water containing 0.1% TEA = 0% to 30%) to obtain E15-E18 as a white solid.

[0712]

[0713] Step 1: Phosphoramidate 44

[0714]

[0715] To a solution of C3 (16 mg, 0.014 mmol, co-evaporated with Py 1 mL×3) in Py (0.5 mL) was added DMOCP (10.4 mg, 0.056 mmol). After stirring for 15 minutes, NIS (4.1 mg, 0.0183 mmol) and morpholine (0.012 mL, 0.141 mmol) were added, and the mixture was stirred for 1 hour, and then aqueous sodium bisulfite solution (0.14%, 1 mL) and sodium bicarbonate (80 mg) were added. The mixture was then extracted with DCM (5 mL×3), dried over anhydrous sodium sulfate, and concentrated. The residue was then stirred in MeCN (0.5 mL) and tert-butylamine (0.5 mL) at room temperature for 15 minutes, and then concentrated. The resulting residue was then co-evaporated with MeCN (1 mL×3) and purified by HPLC to give 86 as a white solid (2.4 mg, 15%). (MS: [M+H] + 1146.2)

[0716] Step 2: E24

[0717]

[0718] Methylamine (33% in EtOH, 0.3 mL) was added to 86 (2.4 mg, 0.0021 mmol). After stirring at room temperature for 16 hours, the mixture was concentrated and the residue was stirred at 35 ° C in a mixture of TEA and TEA·3HF in THF (0.036 mL / 0.018 mL / 0.3 mL) for 18 hours. MeCN (1.0 mL) was then added, and the solid was collected by centrifugation and washed with MeCN (1 mL×2) to give E24 as a white solid (0.6 mg, 38% yield). (MS: [M+H] + 744.0)

[0719]

[0720] Step 1: Borane Phosphate 87

[0721]

[0722] To a solution of C3 (100 mg, 0.088 mmol, co-evaporated with Py 4 mL×3) in Py (3 mL) was added DMOCP (57 mg, 0.337 mmol). After stirring for 15 minutes, BSTFA (0.10 mL, 0.371 mmol) was added dropwise, and the mixture was stirred for 20 minutes, and then borane N,N-diisopropylethylamine complex (0.092 mL, 0.530 mmol) was added. The mixture was then stirred for 3 hours, then concentrated, and purified by silica gel column chromatography (MeOH / DCM=1 / 19 to 1 / 9) to give semi-pure CE-protected 87 as a yellow solid. The semi-pure CE-protected 87 obtained above was stirred in a mixture of MeCN (1 mL) and tert-butylamine (0.5 mL) for 10 minutes, and then concentrated. The residue was then co-evaporated with MeCN (4 mL×3) and purified by reverse phase HPLC (MeCN / water containing 0.1% TEA = 40% to 90%) to give 87 as a white solid (11 mg, 12% yield for two steps). (MS: [M] - 1073.2)

[0723] Step 2: E25

[0724]

[0725] Methylamine (33% in EtOH, 1 mL) was added to 87 (5.7 mg, 0.0053 mmol). After stirring at room temperature for 18 hours, the mixture was concentrated and the residue was stirred in a mixture of TEA (0.08 mL) and TEA·3HF (0.04 mL) in THF (0.5 mL) at 35 ° C for 18 hours. MeCN (1.2 mL) was then added, and the solid was collected by centrifugation and purified by reverse phase HPLC (MeCN / water containing 0.1% TFA = 0% to 20%) to give E25 as a white solid (2.5 mg, 61% yield). (MS: [M] - 671.2)

[0726]

[0727] Step 1: CB1

[0728]

[0729] GB3 (160 mg, 0.25 mmol, co-evaporated with MeCN 1 mL×3) and pyridinium trifluoroacetate (35 mg, 0.39 mmol, co-evaporated with MeCN 1 mL×3) were added in a flask. A solution of A4 (355 mg, 0.36 mmol, co-evaporated with MeCN 1 mL x 3) was added to a solution of MS (500 mg) in MeCN (1 mL) for 30 min. MS (700 mg) was treated with a solution in MeCN (1 mL) for 30 minutes. After stirring at room temperature for 2 hours, TBHP (5.5 M in decane, 0.164 mL, 0.9 mmol) was added, and the mixture was stirred for 30 minutes, and then an aqueous sodium bisulfite solution (33%, 0.15 mL) was added at 0°C. The mixture was then concentrated, and the residue was dissolved in DCM (4.8 mL), followed by the addition of water (0.054 mL) and dichloroacetic acid (6% methylene chloride solution, 4.8 mL). After stirring for 10 min, Py (1.5 mL) was added, the mixture was concentrated, and CB1·Py was purified by silica gel column chromatography (MeOH / DCM=1 / 10 and 1 / 4, containing 1% Py) to give a white solid (213 mg, 66% yield).

[0730] Step 2: Phosphorothioate 88

[0731]

[0732] DMOCP (30 mg, 162.5 μmol) was added to a solution of CB1 (60 mg, 53 μmol) in Py (1 mL). After stirring for 10 minutes, water (0.027 mL) and 3H-1,2-benzodithiol-3-one (13 mg, 0.077 mmol) were added. The mixture was stirred for 5 minutes and then poured into a solution of sodium bicarbonate (210 mg) in water (7.5 mL). After stirring for 5 minutes, the mixture was extracted with EA / ether (1:1, 10 mL×3). The combined organic layers were concentrated to give a yellow solid (100 mg). Tert-butylamine (0.5 mL) was added to a solution of the yellow solid obtained above in MeCN (0.5 mL). After stirring for 10 minutes, the mixture was concentrated and purified by HPLC (MeCN / water containing 0.1% TFA: 50% to 100%) to give two diastereomers of 88. Isomer 1 (7 mg) ([M+H] + 977.0); Isomer 2 (16 mg) (MS: [M+H] + 977.0)

[0733] Step 3: EB1 and EB2

[0734]

[0735] To isomer 1 of 88 (7 mg) was added methylamine (33% in ethanol, 1 mL). After stirring at room temperature for 12 hours, the mixture was concentrated and the residue was dissolved in aqueous TFA solution (3% v / v, 1 mL). After stirring for 2 hours, the mixture was concentrated and purified by HPLC (MeCN / water containing 0.1% TFA, 0% to 45%) to give EB1 as a white solid (2.5 mg, 57% yield). (MS: [M+H] + 689.0)

[0736] To isomer 2 of 63 (16 mg) was added methylamine (33% in ethanol, 2 mL) at 0°C. After stirring at room temperature for 12 hours, the mixture was concentrated, and the residue was co-evaporated with a mixture of Py / TEA (5 mL / 2 mL×3) and then dissolved in Py (0.04 mL). TEA (0.25 mL) and TEA·3HF (0.15 mL) were then added. After stirring at 55°C for 3 hours, acetone (2 mL) was added. The solid (10 mg) was collected by filtration and purified by HPLC (MeCN / water containing 0.1% TFA, 0% to 30%) to give EB2 as a white solid (5 mg, 45% yield). (MS: [M+H] + 689.0, [M–H] – 687.0)

[0737] Example F: Synthesis of EC25

[0738]

[0739] To a solution of E1·TEA salt (10 mg, 0.0114 mmol) in water (0.3 mL) was added 2-chloroacetaldehyde (0.015 mL, 0.118 mmol) and aqueous sodium hydroxide solution (1 M, 0.012 mL, 0.012 mmol). After stirring at 37° C. for 18 hours, the mixture was concentrated and purified by reverse phase HPLC (MeCN / water containing 0.1% TFA = 0% to 30%) to give EC26 as a white solid. (MS: [M] - 697.1)

[0740] The following compounds were prepared essentially by the above method.

[0741]

[0742]

[0743]

[0744]

[0745]

[0746]

[0747]

[0748]

[0749]

[0750]

[0751]

[0752]

[0753]

[0754]

[0755]

[0756]

[0757]

[0758]

[0759]

[0760]

[0761]

[0762]

[0763]

[0764]

[0765]

[0766]

[0767]

[0768]

[0769]

[0770]

[0771]

[0772]

[0773]

[0774]

[0775]

[0776]

[0777]

[0778]

[0779]

[0780]

[0781]

[0782]

[0783]

[0784]

[0785]

[0786]

[0787]

[0788]

[0789]

[0790]

[0791]

[0792]

[0793]

[0794]

[0795]

[0796]

[0797]

[0798]

[0799]

[0800]

[0801]

[0802]

[0803]

[0804]

[0805]

[0806]

[0807]

[0808]

[0809]

[0810]

[0811]

[0812]

[0813] Stereochemical information for representative compounds is given below.

[0814] Table 5. P-configuration of Examples E23 and EB3

[0815]

[0816] Biological tests

[0817] Serial dilutions of cGAMP analog compounds in phosphate-buffered saline (PBS) were incubated with THP1 luciferase reporter cells in 96-well plates at 0.2 × 10 6 / well was mixed in the presence or absence of 1 nM Perfringolysin O (PFO), which promotes compound uptake by forming open channels on the plasma membrane. After 16 hours, 15 μL of medium from each well was transferred to a new plate and luminescence was measured. The fold increase in luminescence compared to PBS-stimulated cells was plotted against the logarithm of the concentration of each compound and the EC was calculated using Graphpad 50 .

[0818] Table 6. Activities of cyclic dinucleotides and analogs

[0819]

[0820]

[0821]

[0822]

[0823] 2'3'-cGAMP can be degraded by ecto-nucleotide pyrophosphatase / phosphodiesterase (ENPP1) present in fetal bovine serum (FBS) (Li et al., 2015, Nat Chem Biol, 11, 235). To test whether the cGAMP analog has improved stability, 5 μL of synthetic cGAMP analog (100 μM stock solution) was incubated with 45 μL FBS in a final volume of 50 μL at 37°C for 1, 2 and 4 hours. At the specified time, 10 μL of the reaction mixture was removed and mixed with 10 μL of phosphate buffered saline (PBS), then heated at 95°C to denature the protein, which was removed by centrifugation at 13000g for 5 minutes. As described above, the supernatant was delivered to the THP1-ISG-luciferase cell line in the presence of PFO to measure the activity of the remaining cGAMP analog. Category A indicates less than 10% loss of activity after 4 hours of incubation, B indicates 10-75% loss of activity after 4 hours of incubation, and C indicates more than 75% loss of activity after 4 hours of incubation.

[0824] Table 7. Stability of cyclic dinucleotides and analogs in fetal bovine serum

[0825]

[0826]

[0827] The following series of prophetic examples are also compounds of the present invention:

[0828]

[0829]

[0830]

[0831]

Claims

1. A compound, wherein the compound is a compound of formula Ic or a pharmaceutically acceptable salt thereof: Formula Ic in Z 12 and Z 15 is N; Z 13 and Z 16 It is CH; Z 14 and Z 17 is N; R 3 is substituted by one or more halogen, thiol, hydroxyl, amino, C 1-6 Alkylamino or di(C 1-6 Alkyl) amino-functionalized C2 alkyl; R 4 It is hydroxyl; R 9 and R 10 Each is independently a hydroxyl group or a thiol group.

2. The compound according to claim 1, wherein R 9 and R 10 It's a hydroxyl group.

3. The compound according to claim 1, wherein R 9 and R 10 It's a thiol group.

4. The compound according to claim 1, wherein the compound has the following structure: or a pharmaceutically acceptable salt thereof.

5. The compound according to claim 1, wherein the compound has the following structure: or a pharmaceutically acceptable salt thereof.

6. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 5 and one or more pharmaceutically acceptable excipients.

7. Use of the pharmaceutical composition according to claim 6 in the preparation of a medicament for inducing or promoting an immune response.

8. Use of the pharmaceutical composition according to claim 6 in the preparation of a medicament for treating cancer.

9. Use of the pharmaceutical composition according to claim 6 in the preparation of a medicament for treating infectious diseases.

10. Use of the pharmaceutical composition according to claim 6 in the preparation of a medicament for treating immune disorders.

11. Use of the pharmaceutical composition according to claim 6 in the preparation of a medicament for treating cancer, wherein the composition is used for administration in combination with an immune checkpoint inhibitor.

12. Use of the pharmaceutical composition according to claim 6 in the preparation of a medicament for treating cancer, wherein the composition is used for administration in combination with radiation.

13. Use of the pharmaceutical composition according to claim 6 in the preparation of a medicament for treating cancer, wherein the composition is used for administration in combination with an anti-tumor agent.

14. An immunogenic composition comprising an antigen or an antigenic composition and a compound according to any one of claims 1 to 5.

15. Use of the immunogenic composition according to claim 14 in the preparation of a medicament for treating or preventing a disease, wherein the immunogenic composition is used for administration to a patient suffering from or susceptible to the disease.

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