MODIFIED CYCLIC DINUCLEOTIDE COMPOUNDS

专利摘要:
the present invention relates to the compounds of formula (i), where base1 and base2 are defined as in claim 1, are modulators of sting.
公开号:BR112019017812A2
申请号:R112019017812-0
申请日:2018-03-16
公开日:2020-03-31
发明作者:Oost Thorsten；Fleck Martin；Kuttruff Christian；Carotta Sebastian
申请人:Boehringer Ingelheim International Gmbh；
IPC主号:

专利说明:

Invention Patent Descriptive Report for MODIFIED CYCLIC DINUCLEOTIDE COMPOUNDS.
Field of the Invention [001] The present invention relates to modified cyclic dinucleotide compounds (CDNs) of formula (I) and a pharmaceutically acceptable salt thereof, which induce the production of cytokines. These modified CDNs are composed of a 3 ', 5' linked blocked nucleic acid (LNA) and a 3 ', 5' 2 'monofluorinated linked nucleotide, both of which comprise a purine nucleobase and a phosphorothioate moiety. In addition, the invention relates to pharmaceutical compositions and combinations comprising said compounds, and their use in methods for the treatment of diseases associated with, or modulated by, STING (Interferon Gene Stimulator). In particular, the pharmaceutical compositions of the invention are suitable for the therapy of inflammation, allergic and autoimmune diseases, infectious diseases, cancer and as vaccine adjuvants.
Background of the Invention [002] The role of the immune system is to protect the body from pathogens and malignant cells. However, viruses and cancer cells find ways to escape the immune system. The purpose of immunotherapies is, therefore, to initiate a specific immune response to the antigen or to reactivate a pre-existing response in certain types of cells of the immune system against pathogenic invaders or cancer cells.
[003] The immune system consists of several specialized strains that can be roughly grouped into two arms, the innate and adaptive immune systems. For a successful immune reaction, strains in both arms must act in harmony. An important role of the innate immune system is to assemble
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2/80 a rapid immune response against pathogens or malignant cells that, unlike the adaptive system, is not antigen-specific and long-lasting. In addition to the direct death of pathogens or transformed cells, the innate immune system also activates and subsequently targets the adaptive immune system. Antigen presenting cells, such as dendritic cells, capture and present the antigens in the form of a major peptide histocompatibility complex (MHC) to T cells in lymphoid tissues. This presentation of antigens together with the secretion of certain cytokines leads to the activation and differentiation of antigen-specific CD4 and CD8 effector cells. The production of type I interferon (IFN) by antigen presenting cells, and other types of cells, is considered a key event in the activation of T cells, as the lack of type I IFN resulted in a reduced T cell-dependent immune response. against viral infections or tumor cells. (Zitvogel et al, Nature Reviews Immunology 15, 405 414, 2015). On the other hand, the presence of a type I IFN signature during cancer therapy is associated with the increased numbers of T cells that infiltrate tumors and the potentially favorable clinical outcome (Sistigu et al, Nature Medicine 20, 1301 - 1309, 2014).
[004] Recent studies in mice have shown that the efficient secretion of type I IFN in the tumor microenvironment and the induction of a T-cell-dependent immune response against cancer cells depends on the presence of the interferon gene-stimulating adapter protein ) (STING, also known as Tmem173, MPYS, MITA, ERIS) (Woo et al, Immunity 41, 5, 830 - 842, 2014; Corrales et al, Cell Reports 11, 1018 - 1030, 2015; Deng et al, Immunity 41, 5, 843 - 852, 2014). The importance of the presence of IFN type I was highlighted by the fact
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3/80 that the removal of STING resulted in reduced levels of IFN type I in the tumor microenvironment and a reduced antitumor effect in several tumor models of mice. On the other hand, the specific activation of STING resulted in an improved immune response of antigen-specific T cells against cancer cells.
[005] STING belongs to the family of nucleic acid sensors and is the adapter for cytosolic DNA signaling. In its basal state, STING exists as a dimer with its N terminal domain anchored in the ER and the C terminal domain residing in the cytosol. Cyclic dinucleotides (CDNs), generated by the cyclic GMP-AMP synthase protein (cGAS), are STING's natural ligands (Ablasser et al, Nature 498, 380 - 384, 2013). The binding of CDNs to STING induces conformational changes that allow the binding and activation of the TANK binding kinase (TBK1) and interferon regulatory factor 3 (IRF3) and the relocation of the ER to perinuclear endosomes (Liu et al, Science 347, Issue 6227, 2630-1 - 2630-14, 2015). Phosphorylation of the IRF3 and NF-kB transcription factor by TBK1 results in the expression of multiple cytokines, including IFN type I.
[006] Given the importance of type I IFN in various malignancies, including viral infections and cancer therapy, strategies that allow specific STING activation are of therapeutic interest.
[007] WO 2014/093936 describes cyclic dinucleotide compounds that have two purine nucleobases and two canonical 3 ', 5' phosphodiester or phosphorothioate moieties and induce STING-dependent cytokine production.
[008] US 7,709,458 describes cyclic dinucleotide compounds that have two purine nucleobases and two canonical 3 ', 5' phosphodiester moieties and can be used to inhibit cancer cell proliferation or to increase cell apoptosis
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Carcinogenic ions, in particular the c-di-GMP symmetrical bacterial CDN. [009] US 7,592,326 describes immunostimulatory cyclic dinucleotide compounds that have two purine nucleobases and two canonical 3 ', 5' phosfordiester moieties, in particular the c-di-GMP symmetrical bacterial CDN.
[0010] WO 2016/096174 and WO 2016/145102 describe cyclic dinucleotide compounds that have two purine nucleobases and two canonical 3 ', 5' phosphodiester or phosphorothioate moieties and induce the production of STING-dependent cytokines.
[0011] Bioorg. Med. Chem. Lett. 18 (2008) 5631-5634 describes the immunostimulatory mono- and bis-phosphorothioate analogs of the c-di-GMP symmetrical bacterial CDN.
[0012] WO 2014/189805 describes cyclic dinucleotide compounds that have two purine nucleobases and at least a 2 ', 5' phosphodiester or non-canonical phosphorothioate moiety and induce STING-dependent cytokine production.
[0013] WO 2015/185565 describes cyclic dinucleotide compounds that have two purine nucleobases, one or two cyclopentane rings instead of tetrahydrofuran ribose rings and a 2 ', 5' non-canonical phosphodiester moiety and modulate STING .
[0014] WO 2016/120305 describes cyclic dinucleotide compounds that have two purine nucleobases, a ribose portion in which 2'-OH is replaced by a 2'-F and a 2 ', 5' phosphodiester portion non-canonical and modulate STING.
[0015] US 2014/0329889, WO 2014/099824, WO 2015/017652, Cell 154, 748-762 (2013) and Molecular Cell 51, 226-235 (2013) describe the cyclic dinucleotide 2'3'- cGAMP ([G (2 ', 5') pA (3 ', 5') p] cyclic) which has two purine nucleobases, a portion of 3 ', 5' canonical phosphodiester and one of 2 ', 5' non-phosphodiester canonical. Non-canonically bound 2'3'-cGAMP binds to the
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Base ²
[0018] where
Base ¹ and Base ² are nucleobases
Human STING with greater affinity than canonically linked 3'3'-cGAMP or symmetrical bacterial c-di-GMP and induces the production of type I interferon.
[0016] Additional cyclic dinucleotides with connectivity of 2 ', 5'-2', 5 'or 2', 5'-3 ', 5' are described as STING agonists in WO 2017/027645 and WO 2017/027646, respectively .
Summary of the Invention [0017] In a first aspect, the present invention relates to a compound of formula (I) $ H * Base ¹ (I) of purine independently selected from the group consisting of purine, adenine, guanine, xanthine and hypoxanthine, linked through their N ⁹ nitrogen atoms, [0020] isoforms, tautomers, stereoisomers, metabolites, prodrugs, solvates, hydrates and their salts.
[0021] In a second aspect, the present invention relates to a pharmaceutical composition comprising one or more compounds of formula (I), as defined above or below, or a pharmaceutically acceptable salt thereof, optionally together with one or more more inert carriers and / or thinners.
[0022] In a third aspect, the present invention relates to a pharmaceutical composition comprising one or more compounds of formula (I), as defined above or below, or a pharmaceutically acceptable salt thereof, and one or more therapeutics
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6/80 additional pharmaceuticals, optionally in conjunction with one or more inert carriers and / or diluents.
[0023] In a fourth aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use as a medicament.
[0024] In a fifth aspect, the present invention relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as a vaccine adjuvant.
[0025] In a sixth aspect, the present invention relates to a method for the treatment of diseases or conditions associated with, or modulated by, STING, particularly for the treatment of inflammation, allergic or autoimmune diseases, infectious diseases or cancer, in a patient who needs it.
[0026] In addition, the present invention relates to the use of one or more of said inhibitors in the manufacture of a medicament for the treatment of diseases or conditions associated with, or modulated by, STING, particularly for the treatment of inflammation, diseases allergic or autoimmune diseases, infectious diseases or cancer, in a patient who needs it.
The present invention also relates to a compound of formula (I), as defined above or below, or a pharmaceutically acceptable salt thereof for use in a method for the treatment of diseases or conditions associated with, or modulated by, , STING, particularly for the treatment of inflammation, allergic or autoimmune diseases, infectious diseases or cancer, in a patient who needs it.
[0028] Other aspects of the present invention will become apparent to the person skilled in the art directly from the preceding and following description and examples.
General Terms and Definitions
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7/80 [0029] Terms not specifically defined in this document should be given the meanings that would be given to them by a person skilled in the art considering the disclosure and context. As used in the specification, however, unless otherwise specified, the following terms have the indicated meaning and the following conventions are respected.
[0030] The terms compound (s) according to this invention, compound (s) of formula (I), compound (s) of the invention and the like indicate the compounds of formula (I) according to the present invention, including those their tautomers, stereoisomers and mixtures thereof and their salts, in particular the pharmaceutically acceptable salt thereof, and the solvates and hydrates of those compounds, including the solvates and hydrates of those tautomers, stereoisomers and salts thereof.
[0031] Unless specifically indicated, throughout the specification and the appended claims, a given chemical formula or name will include tautomers and all stereo, optical and geometric isomers (eg, enantiomers, diastereoisomers, E / Z isomers etc. .) and racemates thereof, as well as mixtures in proportions other than the separate enantiomers, mixtures of diastereoisomers, or mixtures of any of the preceding forms where such isomers and enantiomers exist, as well as salts, including a pharmaceutically acceptable salt thereof and its solvates, such as, for example, hydrates, including solvates of free compounds or solvates of a salt of the compound.
[0032] In the event that a compound of the present invention is represented in the form of a chemical name and as a formula, the formula will prevail in the event of any discrepancy.
[0033] An asterisk can be used in the subformulas to indicate the bond that is linked to the central molecule as defined.
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8/80 [0034] The term substantially pure, as used herein in connection with the compounds of formula (I), refers to a diastereoisomer (Rp, Rp), (Rp, Sp), (Sp, Rp) or (Sp , Sp) which is at least 75% pure in relation to the other possible diastereoisomers in relation to phosphorus atoms. In preferred embodiments, a substantially pure compound of general formula (I) is at least 85% pure, at least 90% pure, at least 95% pure, at least 97% pure, or at least 99% pure.
[0035] The term protecting group, as used in this document and unless otherwise defined, refers to a chemical functional group that is attached to an oxygen, nitrogen or phosphorus atom to prevent further reaction of that atom, or for other purposes . A wide variety of protecting groups are known to those skilled in the art of organic synthesis, and are described, for example, in Protective Groups in Organic Synthesis by T.W. Greene and P.G.M. Wuts, Third Edition, 1999.
[0036] The term pharmaceutically acceptable is used in this document to refer to those compounds, materials, compositions and / or dosage forms that are, within the scope of correct medical judgment, suitable for use in contact with human tissues and animals, without excessive toxicity, irritation, allergic response or other problem or complication, and comply with a reasonable benefit / risk ratio.
[0037] As used herein, the pharmaceutically acceptable salt refers to the derivatives of the described compounds, in which the parent compound is modified by preparing its acid or base salts. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues, such as amines; alkaline, ammonium or organic salts of acidic residues, such as the phosphodiester moieties or
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9/80 phosphorothioate; and the like.
[0038] The terms modulated or modulating, or modulating (m), as used in this document, unless otherwise indicated, refer to the activation of the STING pathway with one or more compounds of the present invention, in this case representing the STING agonists.
[0039] The terms treatment and treating, as used in this document, cover both therapeutic, that is, curative and / or palliative, and preventive, that is, prophylactic treatment.
[0040] Therapeutic treatment refers to the treatment of patients having already developed one or more of the said conditions in the manifest, acute or chronic form. Therapeutic treatment can be symptomatic treatment to alleviate the symptoms of the specific indication or a causal treatment to reverse or partially reverse the conditions of the indication or to stop or slow the progression of the disease. In addition, therapeutic treatment covers treatment over a period, as well as chronic therapy.
[0041] Preventive treatment (prevention, prophylactic treatment) refers to the treatment of patients at risk of developing one or more of said conditions, before the clinical onset of the disease to reduce said risk.
[0042] The terms treatment and treating include the administration of one or more active compounds to prevent or delay the onset of symptoms or complications and to prevent or delay the development of the disease, condition or disorder and / or to eliminate or control the disease, condition or disorder, as well as to relieve symptoms or complications associated with the disease, condition or disorder.
[0043] The term therapeutically effective amount means an amount of a compound of the present invention that (i) treats or prevents the particular disease or condition, (ii) alleviates, improves or eliminates one or more symptoms of the particular disease or condition, or ( iii) pre
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10/80 vine or delays the onset of one or more symptoms of the particular disease or condition described in this document.
[0044] When this invention relates to patients requiring treatment, it mainly relates to treatment in mammals, in particular humans.
Detailed Description of the Invention
Compounds of the Invention [0045] In one aspect of the present invention, it is found that the modified CDNs of formula (I)
Base ²
[0046] where Base ¹ and Base ² are defined as above and below, exhibit favorable binding affinity to human STING and favorable activity in cells that contain different human STING alleles, which may allow reaching a pharmacological efficacy in low doses.
Therefore, the compounds of formula (I), as defined above or below, or a pharmaceutically acceptable salt thereof are expected to be useful in the treatment of diseases or conditions associated with, or modulated by, STING.
[0048] Thus, according to an aspect of the present invention, a compound of formula (I)
Base ²
Base ⁴
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11/80 [0049] where Base ¹ and Base ² are defined as above or below, is provided, as well as isoforms, tautomers, stereoisomers, metabolites, prodrugs, solvates, hydrates and their salts, particularly their physiologically acceptable salts with inorganic or organic bases.
[0050] Some preferred meanings of the Base ¹ and Base ² substituents will be given below. Any and each of these definitions can be combined with each other.
[0051] According to one modality, Base ¹ is selected from the group consisting of purine, adenine, guanine, xanthine and hypoxanthine, linked through their N ⁹ nitrogen atoms.
[0052] According to another modality, Base ¹ is selected from the group consisting of adenine and guanine, linked through their nitrogen atoms N ⁹ .
[0053] According to another modality, Base ¹ is purine, linked through its nitrogen atom N ⁹ .
[0054] According to another modality, Base ¹ is adenine, linked through its N ⁹ nitrogen atom.
[0055] According to another modality, Base ¹ is guanine, linked through its N ⁹ nitrogen atom.
[0056] According to another modality, Base ¹ is xanthine, linked through its nitrogen atom N ⁹ .
[0057] According to another modality, Base ¹ is hypoxanthine, linked through its N ⁹ nitrogen atom.
[0058] According to one modality, Base ² is selected from the group consisting of purine, adenine, guanine, xanthine and hypoxanthine, linked through their N ⁹ nitrogen atoms.
[0059] According to another modality, Base ² is selected from the group consisting of adenine, guanine and hypoxanthine, linked through their N ⁹ nitrogen atoms.
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12/80 [0060] According to another modality, Base ² is purine, linked through its N ⁹ nitrogen atom.
[0061] According to another modality, Base ² is adenine, linked through its nitrogen atom N ⁹ .
[0062] According to another modality, Base ² is guanine, linked through its nitrogen atom N ⁹ .
[0063] According to another modality, Base ² is xanthine, linked through its nitrogen atom N ⁹ .
[0064] According to another modality, Base ² is hypoxanthine, linked through its N ⁹ nitrogen atom.
[0065] According to another embodiment, the compound of formula (I) is the compound (1-1)
[0066] According to another embodiment, the compound of formula (I) is the compound (I-2)
[0067] According to another embodiment, the compound of formula (I) is the compound (I-3)
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[0068] According to another embodiment, the compound of formula (I) is the compound (I-4)
[0069] The compounds of the present invention have chiral phosphorus atoms in the Rp or Sp configuration. All stereoisomers of the compounds of formula (I), (1-1), (I-2), (I-3) and ( I-4), in substantially pure form or as mixtures thereof, are covered by the present invention. The compounds of general formula (I), (1-1), (1-2), (13) and (1-4) such as stereoisomers (Rp, Rp), (Rp, Sp), (Sp, Rp) or (Sp, Sp) substantially pure are preferred.
Preparation [0070] The compounds according to the invention and their intermediates can be obtained using synthetic methods which are known to the person skilled in the art and described in the organic synthesis literature. Preferably, the compounds are obtained analogously to the preparation methods explained more fully below, in particular as described in the experimental section. In some cases, the sequence adopted in the execution of the reaction schemes can be varied. Variants of these reactions that are known to the skilled person, but are not described in detail here, can also be used
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14/80 of. The general procedures for preparing the compounds according to the invention will become apparent to the person skilled in the study of the following methodology. The starting compounds are either commercially available or can be prepared by methods that are described in the literature or in this document, or can be prepared in an analogous or similar manner. Before the reaction is carried out, any corresponding functional groups in the starting compounds can be protected using conventional protecting groups. These protecting groups can be cleaved again at an appropriate stage within the reaction sequence, using methods familiar to the person skilled in the art.
[0071] The CDNs described in this document can be prepared as described in detail below, or by other methods known to those skilled in the art. It will be understood by a person skilled in the art in the art that these schemes are in no way limiting and that variations in detail can be made without departing from the spirit of the present invention.
[0072] CDNs can be obtained by methods described in Chem. Rev. 113, 7354-7401 (2013), Org. Lett., 12, 3269-3271 (2010), Tetrahedron 49, 1115-1132 (1993), WO 2017/0247645, WO 2017/027646, WO 2014/189805, WO 2016/096174, WO 2015/185565, WO 2016/145102 or WO 2016/120305 and in the references cited therein.
[0073] According to another aspect of the present invention, the compounds of formula (I) and their salts can be prepared by the methodology described below.
[0074] Those skilled in the art will recognize that the two portions of phosphorothioate in formula (I) may each exist in the R (Rp) or S (Sp) configuration. The methodology described below can yield up to four diastereoisomers in relation to
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15/80 phosphorus atoms, which can be separated by methods known to the person skilled in the art, for example, by chromatography and / or fractional crystallization, for example, HPLC with suitable solvent systems and columns at different stages of synthesis . In some cases, for example, when a sulfurization step occurs in a diastereoselective method, the methodology described below can preferably give rise to just two diastereoisomers, which can be separated by chromatographic or crystallization methods known to the person who is skilled in the art. different stages of synthesis.
[0075] As mentioned above, the compounds of formula (I) can be converted to salts by methods known to those skilled in the art, particularly for pharmaceutical use in pharmaceutically acceptable salts.
[0076] The compounds according to the invention are also advantageously obtainable using the methods described in the examples that follow, which can also be combined for this purpose with the methods known for reading from the literature.
[0077] Substituents not explicitly specified in the following preparation methods are understood to cover the definitions mentioned above, under the Summary of the Invention.
[0078] A compound of formula (I)
[0079] can be prepared by deprotection of a compound of formula (11-1) or (II-2)
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[0080] where R ¹ means NH containing a suitable protecting group, such as benzoyl, and R ² means H (protected adenine) or [0081] R ¹ means OH and R ² means NH containing a suitable protecting group, such as / so -butyryl or A /, A / -dimethylformamidinyl (protected guanine) or [0082] R ¹ and R ² both mean OH (xanthine) or [0083] R ¹ means OH and R ² means H (hypoxanthine) or [0084] R ¹ and R ² both mean H (purine); and [0085] where [0086] R ³ means NH containing a suitable protecting group, such as benzoyl, and R ⁴ means H (protected adenine) or [0087] R ³ means OH and R ⁴ means NH containing a suitable protecting group , such as / so-butyryl or A /, A / -dimethylformamidinyl (protected guanine) or [0088] R ³ and R ⁴ both mean OH (xanthine) or [0089] R ³ means OH and R ⁴ means H (hypoxanthine) or
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17/80 [0090] R ³ and R ⁴ both mean H (purine).
[0091] For example, a compound of formula (11-1) or (II-2) is dissolved in a suitable mixture, for example, methylamine or aqueous ammonia in methanol or ethanol, and stirred at an appropriate temperature, for example, 20-60Ό, for a suitable period, for example, 1-24 hours.
[0092] A compound of formula (11-1) can be prepared by cyclization and subsequent sulfurization of a compound of formula (III1):
[0093] where R ¹ , R ² , R ³ and R ⁴ are defined as above.
[0094] For example, a compound of formula (111-1) is dissolved in a suitable solvent, for example, pyridine or a mixture of pyridine and dichloromethane, and treated with a suitable binding reagent, for example 2-oxide 2-chloro-5,5-dimethyl-1,3,2dioxaphosphorinane (DMOCP), diphenyl chlorophosphate, pivaloyl chloride or adamantoyl chloride, and stirred at an appropriate temperature, for example, -50Ό to 20Ό, for a suitable period for example, 0.1-2 hours. The cyclization reaction is terminated by treatment with a suitable sulfurizing reagent, for example, 3 / - / - 1,2benzodithiol-3-one or elemental sulfur, and stirred at a suitable temperature, for example, -50Ό to 20Ό, for a suitable period, for example, 0.1-2 hours.
[0095] In a similar way, a compound of formula (II-2) can
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18/80 be prepared by cyclization and subsequent sulfurization of a compound of formula (III-2):
[0096] where R ¹ , R ² , R ³ and R ⁴ are defined as above.
[0097] A compound of formula (111-1) can be prepared by coupling a compound of formula (IV-1) with a compound of formula (V-1):

[0098] where R ¹ , R ² , R ³ and R ⁴ are defined as above.
[0099] For example, a compound of formula (V-1) is dissolved in a suitable solvent, for example, acetonitrile, and is treated with a solution of a compound of formula (IV-1) dissolved in a suitable solvent, for example example, acetonitrile, optionally in the presence of a suitable binding reagent, for example, tetrazole, Activator 42® (activator solution, containing 5- (3,5-bis (trifluormethyl) phenyl) -1 Htetrazole in acetonitrile), pyridinium dichloroacetate or pyridinium trifluoracetate (or mixtures of binding reagents), and stirred at a suitable temperature, for example, 0 - 20Ό, for a suitable period, for example, 0.1-2 hours. The binding reaction is terminated by treatment with a suitable sulfurizing reagent, for example
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19/80 pio, 3 - (((N, N-dimethylaminomethylidene) amino) -3 / 7-1,2,4-dithiazol-3-thione (DDTT) or phenylacetyl disulfide (PADS) or 1,1-dioxide 3 / -7-1,2benzodithiol-3-one (Beaucage reagent), and stirred at a suitable temperature, for example, 0 - 20Ό, for a suitable period, for example, 0.1-2 hours. After evaporation of the solvent, the residue is dissolved in a suitable solvent, for example, a mixture of dichloromethane and water, and treated with a suitable reagent, for example, dichloroacetic acid, and stirred at a suitable temperature, for example, 0 - 20Ό , for a suitable period, for example, 0.1-1 hour.
[00100] A compound of formula (III-2) can be prepared in an analogous manner by coupling a compound of formula (IV-2) with a compound of formula (V-2):

[00101] where R ¹ , R ² , R ³ and R ⁴ are defined as above.
[00102] A compound of formula (IV-1) can be prepared by reacting a compound of formula (V-2), as defined above. For example, a commercially available compound of formula (V-2) is dissolved in a suitable mixture, for example, acetonitrile containing water, and treated with pyridinium trifluoracetate and stirred at a suitable temperature, for example, 0 - 20Ό, for a suitable period, for example, 1-30 minutes. Then the tert-butylamine is added and the mixture is stirred at a suitable temperature, for example 0 - 20Ό, for a suitable period, for example, 0.1-1 hour. The product is isolated by evaporation of the solvent, then I dissolved
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20/80 in a suitable solvent, for example, dichloromethane containing water, and treated with dichloroacetic acid and stirred at an appropriate temperature, for example 0 - 200, for a suitable period, for example, 0.1-1 hour .
[00103] The compounds of general formula I, or their synthetic intermediates, can be resolved into their diastereoisomers taking advantage of their different physical-chemical properties, using methods known perse, for example, chromatography and / or fractional crystallization.
[00104] The compounds according to the invention are also advantageously obtainable using the methods described in the examples which follow, which can also be combined for this purpose with the methods known to the skilled person from the literature.
[00105] The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound which contains an acidic portion by conventional chemical methods. In general, these salts can be prepared by reacting the free acid forms of these compounds with a sufficient amount of the appropriate base in water or in an organic diluent such as ether, ethyl acetate, ethanol, n-propanol, isopropanol, acetone or acetonitrile, or a mixture thereof. Alternatively, reverse phase chromatography of the compounds of the invention (free acid or salt form) employing volatile buffers, such as aqueous solutions of triethylammonium acetate, triethylammonium format, ammonium acetate or ammonium hydrogen carbonate, produces the compounds of the invention as the respective triethylammonium or ammonium salt after freeze-drying / drying. Alternatively, salts can be prepared by ion exchange, for example, by treating aqueous solutions of the compounds of the invention (free acid or salt form) with a cation exchanger.
Pharmacological Activity
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21/80 [00106] The compounds according to the present invention exhibit a binding affinity favorable to human STING. The binding affinity can, for example, be determined by competition binding assay based on the scintillation proximity test (SPA), as described in Nat. Chem. Biol. 10, 1043-1048 (2014). Alternatively, the binding affinity can, for example, be determined by isothermal titration calorimetry (ITC), as described in Molecular Cell 51, 226-235 (2013). Alternatively, the binding affinity can, for example, be determined by surface plasmon resonance (SPR), as described in WO 2016/145102. Alternatively, the binding affinity can, for example, be determined by differential scanning fluorimetry (DSF), as described below.
[00107] The compounds according to the present invention exhibit favorable cellular activity. In vitro cytokine induction can be measured in reporting cell lines, for example, in THP1 cells, as described below. Human STING exists in at least five known variants (WT, HAQ, REF / 232H, AQ, Q / 293Q). To test the activity of the different CDNs on human STING variants, THP1-STING KO cells can be stably transduced with vectors that code for the different STING variants. In addition, in vitro cytokine induction can be measured in primary human PBMCs or in human dendritic cells.
[00108] The compounds according to the present invention exhibit favorable stability in in vitro cell assays, for example, with THP1 cells, Calu-3 cells or human hepatocytes. In addition, the compounds according to the present invention exhibit favorable pharmacokinetic properties (PK). PK properties can be determined in preclinical animal species, for example, mouse, rat, hamster, dog, guinea pig, mini pig, cynomolgus monkey, rhesus monkey. The PK properties of a
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22/80 compound can be described, for example, by the following parameters: Mean residence time (MRT), elimination half-life (ti / 2, that is, the time required for the concentration of the drug to reach half its original value ), volume of distribution (Vd, that is, the apparent volume in which a drug is distributed), area under the curve (AUC, that is, the integral of the concentration-time curve after a single dose), clearance (CL, that is, the drug's purified plasma volume per unit of time), as described in E. Kerns & L. Di (Drug-like properties: concepts, structure design and methods: from ADME to toxicity optimization, Elsevier, 1- ed , 2008). In addition, the compounds according to the present invention exhibit favorable in vivo pharmacological activity, for example, in tumor models MC38, B16, 4T1, Colon26, mouse EMT6 after intratumor application.
[00109] The affinity of favorable binding to human STING in combination with favorable cellular activity and / or favorable PK properties may allow lower doses for pharmacological efficacy. The lower doses have the advantages of lower drug load or drug overload (original drug and its metabolites) for the patient, potentially causing less side effects and lower production costs for the drug. Similarly, the potent activity against human STING variants is advantageous, since it maximizes the chances of inducing the desired pharmacological response in patients with single nucleotide polymorphisms, when a fixed dose is administered.
[00110] The binding of the compounds of the invention to human STING can be demonstrated using the following assay:
DIFFERENTIAL SCANNING FLUORIMETRY DIGITALIZATION (DSF) [00111] Materials:
[00112] PCR plates Hard-Shell® 384 Well-walled wells (Ca
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23/80 n ° ^Q HSP3805R, BIO-RAD) [00113] Microseal®'B 'Adhesive Seals for PCR Plates (Catalog No. ² MSB-1001, BIO-RAD) [00114] SYPRO orange solution in DMSO (SIGMA cat. No. S5692500UL), 5000x concentration [00115] Instrumentation: Reader: CFX384 Real-Time System (Bio-Rad) [00116] Pipetting Robot: HamiltonStarlet [00117] Assay buffer: 20 mM Tris, 150 mM NaCI pH 7, 5 [00118] Target Protein: Human STING (hSTING, residues 155-341, His8 N terminal wild type sequence and VTE dividing site, PM: 23601,5Da) [00119] Protein stock solution: c = 309 μΜ of stock solution in assay buffer [00120] Concentrations in the Final Assay of test compounds: 100 μΜ, 3 μΜ of target protein, 5x SYPR Orange
Test procedure:
[00121] 1) Compound stock solutions and their dilutions were prepared in assay buffer [00122] 2) 5 μΙ of fluorescent dye stock solution (5000x
SYPRO Orange) were mixed with 50 μΙ of target protein (309 μΜ) and 945 μΙ of buffer.
[00123] 3) 2 μΙ of this mixture of protein and dye (25x SYPRO
Orange and 15 μΜ of protein) were added to 8 μΙ of the compound's solution. The final volume was 10 pL.
[00124] 4) Certain well positions were used as a negative control.
[00125] 5) The plates were prepared for duplicate measurement and centrifuged for 2 min at 1000g.
[00126] 6) In the measurement, 160 cycles of 0.5Ό (gradien
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24/80 t and temperatures 15 s / cycle, 15G to 950).
[00127] Data analysis: The dissociation curves were processed in Bio-Rad CFX Manager. The peak type was set to negative. In the case of Examples 1.1,2.1,3.1 and 4.1, at least two measurements of Tm were averaged. The changes in Tm (thermal displacement) are shown in table 1.
Table 1: hSTING binding as determined by differential scanning fluorimetry
Example HSTING Tm offset [O] 1.1 27.6 1.2 21.0 2.1 30.3 2.2 21.8 3.1 30.7 3.2 22.4 3.3 21.6 3.4 10.9 4.1 30.0 4.2 21.4 Example 5 Compound 23 in WO 2016/145102 21.1
[00128] Surprisingly, it has been found that the compounds according to the present invention exhibit significantly improved binding compared to analogues, where the LNA portion is replaced by a ribose portion, for example, Examples 1.1.2.1, 3.1 and 4.1 of the LNA exhibit significantly greater thermal shifts, i.e. improved binding, compared to Example 5 / Ribose Compound 23 (3'3'-RR- (2'FG) (A)) in WO 2016/145102 . The improved binding of the compounds according to the present invention, in the aforementioned differential scanning fluorimetry (DSF) assay, was also confirmed in a surface plasmon resonance (SPR) assay.
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25/80 [00129] The cellular activity of the compounds of the invention can be demonstrated using the following THP1 assay in vitro:
CYTOKINES INDUCTION IN VITRO [00130] The cytokine-inducing activities of the compounds according to the present invention have been demonstrated using a THP1 reporter cell line.
[00131] Activation of the STING protein expressed in THP1 cells results in an increase in interferon production. Through the stable integration of a reporting construct of SEAP (secreted embryonic alkaline phosphatase) inducible by the interferon regulating factor (IRF), the functional interferon signaling pathway can be monitored. Using Invivogen's QUANTI-Blue® colorimetric enzyme assay and a suitable optical density (DO) reader, SEAP activity can be detected and quantified. This technique can be used to characterize the pharmacological modification of the STING protein.
[00132] Measurements of SEAP activity were performed on THP1-Blue ISG cells that stably express the human STING protein and the SEAP reporting construct inducible by IRF. The cells were cultured for expansion in RPMI1640 medium with 10% fetal calf serum, 50 pg / ml Penicillin-Streptomycin, 100 pg / ml Zeocin and 100 pg / ml Normocin in a 370 incubator, 95% moisture and 5% CO 2. The cells ready for the test were stored as frozen stocks.
[00133] In preparation for the assay, the cells were thawed in medium without Zeocin / Normocin and were distributed on the assay plates with a density of 15000 cells / 15 μΙ_ per well. The compounds were prepared by a serial dilution of 8 or 16 points in 50% aqueous DMSO and a final dilution step in the medium to ensure a final concentration of DMSO of 0.5% in the assay. 5 μΙ_ of the diluted compounds plus 5 μΙ_ of medium were added to the
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26/80 plates, followed by an incubation for 24 hours at 37Ό.
[00134] On the day of the test, 75 μΙ_ per well of Quanti-Blue reagent was added to all wells on the plate and the plate was incubated for another 30 minutes at 37Ό. The 620 nm OD was measured on the EnVision reader (PerkinElmer).
[00135] EC50 values and Hill gradients were derived from four adaptations of the 8 or 16 point nonlinear curve with 0 Megalab software (Boehringer Ingelheim), using the OD at 620 nM. See Table 2.
Table 2: Cellular activity in the THP1-Blue ISG cell assay
Example EC ₅₀ [μΜ] 1.1 0.11 1.2 0.06 2.1 0.14 2.2 0.05 3.1 0.15 3.2 0.07 3.3 0.13 3.4 2.0 4.1 0.17 4.2 0.11
[00136] Several single nucleotide polymorphisms have been identified in the human STING gene, which can affect the response to cyclic dinucleotides. To determine the activity of the compounds of the invention, THP1-Blue ISG reporting cell lines were generated expressing the different variants of human STING. To do this, the endogenous human STING was first removed using the CRISPR / CAS9 system: THP1-Blue ISG cells were electroporated with the ALL-IN-ONE CRISPR plasmids targeting the STING gene (acquired from Sigma encoding 0 gRNA and the GFP as a reporting gene for successful transduction). Positive cells for
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27/80
GFP were then separated 24 hours after transfection and expanded. The cells were then dispersed in semi-solid methocel medium to allow isolations of individual cell clones. The clones were then examined for sensitivity to cGAMP using the Quanti-blue reporter assay. The unresponsive clones were subsequently analyzed for loss of STING by western blotting and sequencing of the STING locus.
[00137] For overexpression of human STING variants, a confirmed THP1-Blue ISG hSTING KO clone was transduced with individual retroviral plasmids (MSCV-ires-GFP-Blasti) encoding the allele variants of hSTING (WT, HAQ, R232H, AQ and R293Q), respectively. The transduced cells were separated according to the different levels of GFP fluorescence and the expression of the STING allele was analyzed by western blot. Populations expressing ectopic STING protein (WT, HAQ, R232H, AQ and R293Q) at levels comparable to endogenous STING levels from parental, unmodified THP1-Blue ISG cell lines were selected and used to characterize the compounds. Surprisingly, it was found that the compounds according to the present invention exhibit very potent cell activity in all five of the variant cell lines above, for example, Examples 2.1, 3.1 and 4.1 exhibit EC50 ^ 1 μΜ values in the variant cell line WT, HAQ, R232H, AQ and R293Q, respectively, indicating absence of differences / selectivity of pronounced variants. The observed cell activity is dependent on STING, since no activity was observed in a THP1 cell line, where the human STING was removed.
[00138] The cell stability of the compounds of the invention was determined as follows: The compound was dissolved in cell culture medium (MEM supplemented with 10% FCS, 1% amino acid
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28/80 non-essential acids and 1% pyruvate) to a final concentration of 10 μΜ and incubated with the human lung epithelial cell line Calu-3 (60,000 cells / well in a 24-well plate) for up to 24 h. Samples of cell culture supernatants were obtained at 1, 6, 24 h and quantified by LC-MS / MS.
Treatment Methods [00139] In another aspect of the present invention, it appears that the compounds of formula (I) or a pharmaceutically acceptable salt thereof may be useful for the treatment of diseases or conditions where the modulation of STING is of benefit therapeutic. In addition, due to their activity, the compounds of the present invention are suitable as vaccine adjuvants.
[00140] Diseases and conditions associated with, or modulated by, STING include, but are not limited to, inflammation, allergic or autoimmune diseases, for example, allergic rhinitis or asthma, infectious diseases or cancer.
[00141] Autoimmune diseases include, but are not limited to, systemic lupus erythematosus, psoriasis, insulin-dependent diabetes mellitus (IDDM), dermatomyositis and Sjogren's syndrome (SS).
[00142] Inflammation represents a group of vascular, cellular and neurological responses to trauma. Inflammation can be characterized as the movement of inflammatory cells, such as monocytes, neutrophils and granulocytes, in the tissues. This is usually associated with reduced endothelial barrier function and tissue edema. The inflammation can be classified as acute or chronic. Acute inflammation is the body's initial response to harmful stimuli and is achieved by the increased movement of plasma and leukocytes from the blood to the injured tissues. A cascade of biochemical event spreads and the inflammatory response matures, involving the local vascular system, the immune system and several cells within the injured tissue.
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29/80
Prolonged inflammation, known as chronic inflammation, leads to a progressive change in the type of cells that are present at the site of the inflammation and is characterized by simultaneous destruction and healing of the inflammatory process tissue.
[00143] When it occurs as part of an immune response to infection or as an acute response to trauma, inflammation can be beneficial and is usually self-limiting. However, inflammation can be harmful under several conditions. This includes the production of excessive inflammation in response to infectious agents, which can lead to significant organ damage and death (for example, in the case of sepsis). In addition, chronic inflammation is generally harmful and is the cause of numerous chronic diseases, causing severe and irreversible tissue damage. In such situations, the immune response is often directed against the tissues themselves (autoimmunity), although chronic responses to foreign entities can also lead to direct damage to the tissues themselves. The purpose of anti-inflammatory therapy is, therefore, to reduce this inflammation, inhibit autoimmunity when present, and allow the physiological process or tissue healing and restoration to progress.
[00144] The compounds of the invention can be used to treat inflammation of any tissue and organs in the body, including musculoskeletal inflammation, vascular inflammation, neural inflammation, inflammation of the digestive system, eye inflammation, inflammation of the reproductive system and other inflammations, as exemplified below.
[00145] Musculoskeletal inflammation refers to any inflammatory condition of the musculoskeletal system, particularly conditions that affect the joints of the skeleton, including the joints of the hand, wrist, elbow, shoulder, jaw, spine, neck, hip, knee, ankle and foot, and conditions that affect the tissues that connect muscles to bones, such as tendons. The examples
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30/80 peaks of musculoskeletal inflammation that can be treated with the compounds of the invention include arthritis (including, for example, osteoarthritis, rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, acute and chronic infectious arthritis, associated arthritis gout and pseudogout and juvenile idiopathic arthritis), tendonitis, synovitis, tenosynovitis, bursitis, fibrositis (fibromyalgia), epicondylitis, myositis and osteitis (including, for example, Paget's disease, osteitis and cystic fibrous osteitis). Eye inflammation refers to the inflammation of any structure in the eye, including the eyelids. Examples of ocular inflammation that can be treated with the compounds of the invention include blepharitis, blepharocalasia, conjunctivitis, dacrioadenitis, keratitis, dry keratoconjunctivitis (dry eye), scleritis, trichiasis and uveitis. Examples of inflammation of the nervous system that can be treated with the compounds of the invention include encephalitis, Guillain-Barre syndrome, meningitis, neuromyotonia, narcolepsy, multiple sclerosis, myelitis and schizophrenia.
[00146] Examples of inflammation of the vasculature or lymphatic system that can be treated with the compounds of the invention include arthrosclerosis, arthritis, phlebitis, vasculitis and lymphangitis.
[00147] Examples of inflammatory conditions of the digestive system that can be treated with the compounds of the invention include cholangitis, cholecystitis, enteritis, enterocolitis, gastritis, gastroenteritis, inflammatory bowel disease (such as Crohn and ulcerative colitis), ileitis and proctitis.
[00148] Examples of inflammatory conditions of the reproductive system that can be treated with the compounds of the invention include cervicitis, chorioamnionitis, endometritis, epididymitis, omphalitis, oophoritis, orchitis, salpingitis, tube abscess. ovarian, urethritis, vaginitis, vulvitis and vulvodynia.
[00149] Agents can be used to treat au conditions
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31/80 toimunes having an inflammatory component. Such conditions include universal acute disseminated alopecia, Behcet's disease, Chagas' disease, chronic fatigue syndrome, dysautonomia, encephalomyelitis, ankylosing spondylitis, aplastic anemia, suppurative hidradenitis, autoimmune hepatitis, autoimmune oophoritis , celiac disease, Crohn's disease, type 1 diabetes mellitus, giant cell arteritis, goodpasture syndrome, Graves' disease, Guillain-Barre syndrome, Hashimoto's disease, HenochSchonlein's purple, Kawasaki disease, lupus erythematosus, microscopic colitis, microscopic polyarteritis, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, opsoclone myoclonus syndrome, optic neuritis, ord thyroiditis, pemphigus, polyarteritis nodosa, polymyalgia, rheumatoid arthritis, Reiter's syndrome, Sjogren's syndrome, temporal arteritis, Wegener's granulomatosis, hot autoimmune hemolytic anemia, interstitial cystitis, liver disease yme, morphea, psoriasis, sarcoidosis, scleroderma, ulcerative colitis and vitiligo.
[00150] The agents can be used to treat hypersensitivity diseases mediated by T cells having an inflammatory component. Such conditions include contact hypersensitivity, contact dermatitis (including due to poison ivy), hives, skin allergies, respiratory allergies (hay fever, allergic rhinitis) and gluten-sensitive enteropathy (Celiac disease) .
[00151] Other inflammatory conditions that can be treated with the agents include, for example, appendicitis, dermatitis, dermatomyositis, endocarditis, fibrositis, gingivitis, glossitis, hepatitis, suppurative hidradenitis, iritis, laryngitis, mastitis, myocarditis, nephritis, otitis, pancreatitis, parotitis, percarditis, peritonitis, pharyngitis, pleuritis, pneumonitis, prostatitis, pyelonephritis and stomatitis, transplant rejection ( involving organs like kidney,
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32/80 liver, heart, lung, pancreas (eg islet cells), bone marrow, cornea, small intestine, skin allografts, skin homografts, and heart valve xenografts, serum disease, and graft vs disease . host), acute pancreatitis, chronic pancreatitis, acute respiratory distress syndrome, Sexary syndrome, congenital adrenal hyperplasia, non-suppurative thyroiditis, cancer-associated hypercalcemia, pemphigus, herpetiform bullous dermatitis, multiform erythema severe, exfoliative dermatitis, seborrheic dermatitis, seasonal or perennial allergic rhinitis, bronchial asthma, contact dermatitis, atopic dermatitis, drug hypersensitivity reactions, allergic conjunctivitis, keratitis, ophthalmic herpes zoster, a iritis and oiridocyclitis, chorioretinitis, optic neuritis, symptomatic sarcoidosis, fulminant or disseminated pulmonary tuberculosis chemotherapy, idiopathic thrombocytopenic purpura in adults, secondary thrombocytopenia in adults, acquired hemolytic anemia (autoimmune), leukemia and lymphomas in adults, acute childhood leukemia, regional enteritis, autoimmune vasculitis, multiple sclerosis, obstructive pulmonary disease chronic use, rejection of solid organ transplantation, sepsis. Preferred treatments include transplant rejection treatment, rheumatoid arthritis, psoriatic arthritis, multiple sclerosis, Type 1 diabetes, asthma, inflammatory bowel disease, systemic lupus erythematosus, psoriasis, chronic lung disease and inflammation accompanying infectious diseases (for example, sepsis).
[00152] In one aspect, the disease or condition to be treated using the compounds of the invention is cancer. Examples of cancerous diseases and conditions in which the compounds of formula (I), or their pharmaceutically acceptable salts or solvates, may have potentially beneficial antitumor effects include, but are not limited to, cancers of the lung, bone, pancreas, skin, head,
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33/80 neck, uterus, ovaries, stomach, colon, breast, ovary, esophagus, small intestine, intestine, endocrine system, thyroid gland, parathyroid gland, adrenal gland, urethra, prostate, penis, testicles, ureter, bladder, kidney or liver; urothelial cancer; rectal cancer; cancer of the anal region; carcinomas of the fallopian tubes, endometrium, cervix, vagina, vulva, renal pelvis, renal cell; soft tissue sarcoma; myxoma; rhabdomyoma; fibroma; lipoma; teratoma; cholangiocarcinoma; hepatoblastoma; angiosarcoma; hemangioma; hepatoma; fibrosarcoma; chondrosarcoma; myeloma; chronic or acute leukemia; lymphocytic lymphomas; primary CNS lymphoma; CNS neoplasms; tumors of the spine axis; squamous cell carcinomas; synovial sarcoma; malignant pleural mesotheliomas; brain stem glioma; pituitary adenoma; bronchial adenoma; chondromatous hanlartoma; mesothelioma; Hodgkin's disease or a combination of one or more of the previous cancers.
[00153] Preferred cancers, which can be treated with the compounds according to the invention, are cancers of the skin, lung, liver, colon, brain, breast, ovary, prostate, cancers of the pancreas, kidney, stomach, head , neck, skin and urothelial, as well as lymphoma and leukemia.
[00154] The new compounds can be used for the prevention, short-term or long-term treatment of the aforementioned diseases, optionally also in combination with surgery, radiotherapy or other prior art compounds, such as, for example, substances cytostatic or cytotoxic, cell proliferation inhibitors, antiangiogenic substances, steroids or antibodies.
[00155] In their role as adjuvants, in certain embodiments, the present compounds and compositions can be used as adjuvants in a therapeutic or prophylactic strategy employing vaci
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34/80 in. Thus, the substantially pure CDNs of the present invention, or the prodrugs or a pharmaceutically acceptable salt thereof, can be used in conjunction with one or more selected vaccines to stimulate an immune response to one or more predetermined antigens. The substantially pure CDNs of the present invention, or the prodrugs or a pharmaceutically acceptable salt thereof, can be provided together with, or in addition to, such vaccines.
[00156] Such vaccine (s) may comprise inactivated or attenuated bacteria or viruses comprising the antigens of interest, the purified antigens, the live viral or bacterial release vectors, recombinantly engineered to express and / or secrete the antigens, antigen presenting cell vectors (APC) comprising cells that are loaded with the antigens or transfected with a composition comprising a nucleic acid encoding the antigens, the liposomal antigen release transporters or the pure nucleic acid vectors that encode antigens. This list is not intended to be limiting. As an example, such vaccine (s) may also comprise an inactivated tumor cell that expresses and secretes one or more of GM-CSF, CCL20, CCL3, IL-12p70, FLT-3 linker cytokines.
[00157] The dose range of the compounds of general formula (I) applicable per day is normally from 0.00001 to 10 mg per kg of body weight, for example, from 0.00001 to 1 mg per kg of body weight of the patient . Each dosage unit can conveniently contain from 0.001 to 1000 mg, for example, from 0.001 to 100 mg.
[00158] The therapeutically effective amount or the actual therapeutic dosage will, of course, depend on factors known to those skilled in the art, such as age and weight of the patient, route of administration and severity of the disease. In any case, the compound
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35/80 or the composition will be administered in dosages and in a manner that allows a therapeutically effective amount to be administered based on the patient's unique condition.
[00159] The compounds, compositions, including any combinations with one or more additional therapeutic agents, according to the invention, can be administered via mucosal route (e.g., oral, sublingual, vaginal, nasal, cervical etc.), intratumoral , peritumoral, transdermal, inhalation or parenteral (for example, subcutaneous, intravenous, intramuscular, intra-arterial, intradermal, intrathecal and epidural administrations). Of the possible methods of administration, intratumoral, peritumoral, subcutaneous or intravenous administration is preferred.
[00160] The compounds of the present invention exhibit several advantages, such as favorable binding affinity to human STING, favorable cellular activity, i.e., in cells containing different human STING alleles, favorable stability in cellular assays.
[00161] Thus, in an additional aspect, the invention provides new compounds of formula (I), including a pharmaceutically acceptable salt thereof, which induce the production of cytokines in a STING-dependent manner in vitro and / or in vivo and have pharmacological and pharmacokinetic properties suitable for use in therapy, that is, for use as medicines.
[00162] In a further aspect, the invention provides new compounds of formula (I), including a pharmaceutically acceptable salt thereof, for use in a method for the treatment of a disease or disease associated with, or modulated by, STING.
[00163] In a further aspect, the invention provides new compounds of formula (I), or a pharmaceutically acceptable salt thereof, for the treatment of inflammation, allergic or autoimmune diseases, for example, allergic rhinitis or asthma, for the treatment of
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36/80 infectious or cancer diseases, or for use as vaccine adjuvants.
[00164] In another aspect, the present invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use in the treatment of a disease or condition where the modulation of STING is beneficial.
[00165] In a further aspect, the present invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use in the treatment of inflammation, allergic or autoimmune diseases, for example , allergic rhinitis or asthma, for the treatment of infectious diseases or cancer.
[00166] Consequently, the present invention relates to the compounds of formula (I) as a medicament.
[00167] Furthermore, the present invention relates to the use of a compound of formula (I) in a method for the treatment of diseases or conditions associated with, or modulated by, STING in a patient, preferably in a human .
[00168] Furthermore, the present invention relates to the use of a compound of formula (I) in a method for the treatment of inflammation, allergic or autoimmune diseases, for example, allergic rhinitis or asthma, for the treatment of infectious diseases or cancer.
[00169] In yet another aspect, the present invention relates to a method for treating a disease or condition associated with, or modulated by, STING in a mammal, which includes the step of administering to a patient, preferably a being in need of such treatment, a therapeutically effective amount of a compound or a pharmaceutical composition of the present invention.
[00170] In a further aspect, the invention provides a method
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37/80 all for the treatment of a disease or condition associated with, or modulated by, STING in a patient, which comprises administering a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, to the patient .
[00171] In a further aspect, the invention provides a method for the treatment of inflammation, allergic or autoimmune diseases, for example, allergic rhinitis or asthma, for the treatment of infectious diseases or cancer, in a patient who needs it, comprising administering a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, to the patient.
[00172] In a related aspect, the present invention relates to methods of inducing, stimulating or assisting an immune response in a patient. These methods comprise administering the substantially pure CDNs of the present invention, or the prodrugs or a pharmaceutically acceptable salt thereof, to the patient. [00173] In a further aspect, the invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, for the manufacture of an immunogenic composition comprising an antigen or antigen composition, for the treatment or the prevention of a disease.
[00174] In another aspect, the invention provides a method of treating or preventing a disease, comprising administering to a human patient suffering from, or susceptible to, a disease of an immunogenic composition comprising an antigen or antigen composition and a compound of formula (I), or a pharmaceutically acceptable salt thereof.
[00175] In a further aspect, the invention provides a vaccine composition comprising an antigen or an antigen composition and a compound of formula (I), or a pharmaceutical salt
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38/80 of it, for use in the treatment or prevention of a disease.
[00176] In a further aspect, the invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, for the manufacture of a vaccine composition comprising an antigen or an antigen composition, for the treatment or prevention of a disease.
[00177] In another aspect, the invention provides a method of treating or preventing a disease, comprising administering to a human patient suffering from, or susceptible to, a vaccine composition comprising an antigen or antigen composition and a compound of formula (I), or a pharmaceutically acceptable salt thereof.
Pharmaceutical Compositions [00178] In another aspect of the present invention, it appears that pharmaceutical compositions of the aforementioned compounds can be formulated that are suitable for the administration of therapeutically effective amounts of said inhibitors for the treatment of diseases or conditions associated with, or modulated by, STING.
[00179] For the purposes of this disclosure, pharmaceutical compositions can be administered through a variety of means, including non-parentally, parentally, by inhalation spray, topically or rectally in formulations containing pharmaceutically acceptable carriers, adjuvants and vehicles. The intratumoral (directly in the tumor mass) or peritumoral (around the tumor mass) administration of the compounds of the present invention can directly activate the DC that infiltrates locally, directly promote the apoptosis of tumor cells or sensitize the tumor cells to cytotoxic agents.
[00180] The pharmaceutical compositions of the disclosure may be
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39/80 in the form of a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known technique using suitable dispersing or wetting agents and suspending agents which are mentioned above or below. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic, parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol, or prepared as a lyophilized powder. Among the acceptable carriers and solvents that can be used are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils can conventionally be used as a solvent or suspending medium. For this purpose, any soft fixed oil can be used, including synthetic mono or diglycerides. In addition, fatty acids, such as oleic acid, can also be used in the preparation of injectables.
[00181] Formulations suitable for topical administration in the mouth include expectorant lozenges comprising the active ingredient in a flavored base, usually sucrose and acacia or tragacanth; tablets comprising the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and acacia; and mouthwash liquids comprising the active ingredient in a suitable liquid carrier.
[00182] Formulations suitable for vaginal administration may be presented as pessaries, absorbent tampons, creams, gels, pastes, foams or spray formulations containing, in addition to the active ingredient, such carriers which are known in the art to be suitable.
[00183] Formulations suitable for parenteral administration include solutions for sterile isotonic, aqueous and non-aqueous injections, which may contain antioxidants, buffers, bacteriostats and
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40/80 solutes that make the formulation isotonic with the intended recipient's blood; and sterile aqueous and non-aqueous suspensions which may include suspending agents and thickening agents. The formulations can be presented in sealed single-dose or multiple-dose containers, for example, ampoules and small vials, and can be stored in a lyophilized (desiccated) condition, requiring only the addition of the sterile liquid carrier, for example, water for injections. , immediately before use. Injection solutions and suspensions can be prepared from sterile powders, granules and tablets of the type previously described.
Thus, in accordance with another aspect of the present invention, pharmaceutical compositions are provided comprising one or more compounds of formula (I), or a pharmaceutically acceptable salt thereof, optionally together with one or more inert carriers and / or diluents .
[00185] Furthermore, the present invention relates to the use of a pharmaceutical composition according to this invention for the treatment of diseases or conditions associated with, or modulated by, STING in a patient, preferably in a human.
[00186] In accordance with an embodiment of the second aspect of the present invention, there is provided a pharmaceutical composition comprising one or more of the compounds mentioned above, or a pharmaceutically acceptable salt thereof, optionally together with one or more carriers and / or diluents inert, for use in a method for the treatment of diseases or conditions associated with, or modulated by, STING.
[00187] According to another embodiment, a vaccine is provided comprising one or more compounds of formula (I), or a pharmaceutically acceptable salt thereof.
[00188] In a further aspect, the invention provides an adjuvant
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41/80 vaccine front comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof.
[00189] In a further aspect, the invention provides an immunogenic composition comprising an antigen or antigen composition and a compound of formula (I) or a pharmaceutically acceptable salt thereof.
[00190] In a further aspect, the invention provides an immunogenic composition comprising an antigen or antigen composition and a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use in the treatment or prevention of a disease.
[00191] According to another embodiment, a pharmaceutical composition is provided comprising one or more compounds of formula (I), or a pharmaceutically acceptable salt thereof, and one or more additional therapeutic agents, optionally together with one or more carriers and / or inert diluents. Preferably, this composition comprises a compound of formula (I), or a pharmaceutically acceptable salt thereof, and one or more additional therapeutic agents.
Combination Therapy [00192] The compounds of the invention can be used alone or can be combined with pharmaceutically acceptable excipients, in an amount sufficient to induce, modify or stimulate an appropriate immune response. The immune response can comprise, without limitation, a specific immune response, a non-specific immune response, an both specific and non-specific response, an innate response, a primary immune response, an adaptive immunity, a secondary immune response, an immune response of memory, an activation of immune cells, a proliferation of immune cells, a
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42/80 differentiation of immune cells and an expression of cytokines. In certain embodiments, the compounds and their compositions described in this document are administered in combination with one or more additional compositions, including vaccines designed to stimulate an immune response to one or more predetermined antigens; adjuvants; antagonists of the CTLA-4 and PD-1 pathway, lipids, liposomes, chemotherapeutic agents, immunomodulatory cell lines, etc.
[00193] The compounds and their compositions described in this document can be administered before, after and / or simultaneously with an additional therapeutic or prophylactic composition or modality. These include, without limitation, co-stimulating molecule B7, interleukin-2, interferon-g, GM-CSF, CTLA-4 antagonists, OX-40 / OX-40 ligand, CD40 / CD40 ligand, sargramostim, levamisole, virus vaccinia, Calmette-Guerin bacillus (BCG), liposomes, alum, Freund's complete or incomplete adjuvant, detoxified endotoxins, mineral oils, surfactant substances, such as lipolecithin, pluronic polyols, polyanions, peptides and oil or hydrocarbon emulsions. Transporters for inducing a T cell immune response that preferentially stimulate a cytolytic T cell response versus an antibody response are preferred, although those that stimulate both types of response can be used as well. In cases where the agent is a polypeptide, the polypeptide itself or a polynucleotide that encodes the polypeptide can be administered. The carrier can be a cell, such as an antigen presenting cell (APC) or a dendritic cell. Antigen presenting cells include such cell types as macrophages, dendritic cells and B cells. Other professional antigen presenting cells include monocytes, marginal zone Kupffer cells, microglia, Lange cells
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43/80 rhans, interdigitation dendritic cells, follicular dendritic cells and T cells. Optional antigen presenting cells can also be used. Examples of facultative antigen presenting cells include astrocytes, follicular cells, endothelium and fibroblasts. The carrier can be a bacterial cell that is transformed to express the polypeptide or to release a polynucleotide that is subsequently expressed in the cells of the vaccinated individual. Adjuvants, such as aluminum hydroxide or aluminum phosphate, can be added to increase the vaccine's ability to elicit, enhance or prolong an immune response. Additional materials, such as cytokines, chemokines and bacterial nucleic acid sequences, such as CpG, a Toll-like receptor agonist (TLR) 9, as well as additional agonists for TLR 2, TLR 4, TLR 5, TLR 7, TLR 8, TLR9, including lipoprotein, LPS, monophosphoryl lipid A, lipoteic acid, imiquimod, resiquimod, and agonists of the retinoic acid-inducible I (RIG-I) gene, such as poly 1: 0, used separately or in combination with the described compositions, they are also potential adjuvants. Other representative examples of adjuvants include the synthetic QS-21 adjuvant comprising a homogeneous saponin, purified from the bark of Quillaja saponaria and Corynebacterium parvum (McCune et al., Cancer, 1979; 43: 1619).
[00194] Methods for co-administration with an additional therapeutic agent are well known in the art (Hardman, et al. (Eds.) (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10th ed., McGraw-Hill, New York, NY; Poole and Peterson (eds.) (2001) Pharmacotherapeutics for Advanced Practiced Practical Approach, Lippincott, Williams & Wilkins, Phila., PA; Chabner and Longo (eds.) (2001) Cancer Chemotherapy and Biotherapy, Lippincott, Williams & Wilkins, Phila, PA). In general, coadministration or admiPetition 870190083676, of 08/27/2019, p. 58/103
44/80 joint administration indicates the treatment of a patient with two or more agents, where the agents can be administered simultaneously or at different times. For example, such agents can be administered to a single patient as separate administrations, which can be essentially at the same time or at different times, and which can be by the same route or different routes of administration. Such agents can be administered to a single patient in the same administration (for example, the same formulation) in such a way that they are administered at the same time, by the same route of administration.
[00195] Due to the adjuvant properties of the compounds of the present invention, their use can also be combined with other therapeutic modalities, including other vaccines, adjuvants, antigen, antibodies and immunological modulators. Examples are provided below.
Adjuvants [00196] In addition to the compounds of the present invention and their compositions described in this document, the compositions or methods of the present invention may also comprise one or more additional substances which, due to their nature, may act to stimulate or otherwise use the immune system to respond to cancer antigens present on the tumor target cell (s). Such adjuvants include, but are not limited to, lipids, liposomes, inactivated bacteria that induce innate immunity (for example, inactivated or attenuated Listeria monocytogenes), compositions that mediate innate immune activation through Toll-like Receptors (TLRs), receptors (NOD) -like (NLRs), Receptors (RIG) -l-like (RLRs) based on retinoic acid-inducible gene, type C lectin receptors (CLRs) and / or molecular patterns associated with pathogens (PAMPS). Examples of PAMPs include lipoprote
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45/80 ines, lipopolypeptides, peptidoglycans, zymosan, lipopolysaccharide, neisseria porins, flagellin, profilin, galactoceramide, muramyl dipeptide. Peptidoglycans, lipoproteins and lipoteic acids are components of the Gram-positive cell wall. Lipopolysaccharides are expressed by most bacteria, with MPL being an example. Flagellin refers to the structural component of bacterial flagella that is secreted by pathogenic and commensal bacteria. Galactosylceramide is an activator of natural T killer cells (NKT). Muramyl dipeptide is a bioactive peptidoglycan motif common to all bacteria.
Immune Control Point Inhibitors [00197] The compounds of the present invention can be used in combination with an immune control point inhibitor, such as an immune control point inhibitor selected from the group consisting of an antagonist of the CTLA-4 pathway. , an antagonist of the PD-1 pathway, an antagonist of the Tim-3 pathway, an antagonist of the Vista pathway, an antagonist of the BTLA pathway, an antagonist of the LAG3 pathway or an antagonist of the TIGIT pathway. In some embodiments, the immunological control point inhibitor is selected from the group consisting of an anti-CTLA-4 antibody, an anti-PD-1 antibody, an anti-Tim-3 antibody, an antibody, an anti-Vista, an anti-BTLA antibody, an anti-LAG-3 antibody or an anti-TIGIT antibody.
[00198] The compounds of the present invention can be used in combination with antagonists of the CTLA-4 pathway. In some embodiments, the combination is used to treat a solid tumor or haematological malignancy. CTLA-4 is considered to be an important negative regulator of the adaptive immune response. Activated T cells suppress CTLA-4, which binds CD80 and CD86 on antigen presenting cells with greater affinity than CD28, thereby inhibiting T cell stimulation, gene expression
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46/80 of IL-2 and proliferation of T cells. The anti-tumor effects of CTLA4 blockade have been observed in rat models of colon carcinoma, metastatic prostate cancer and metastatic melanoma. In some embodiments, the CTLA-4 pathway antagonist is an anti-CTLA-4 antibody molecule selected from the group consisting of tremelimumab and ipilimumab.
[00199] ipilimumab (an antibody to CTLA-4, also known as MDX-010, CAS No. 477202-00-9) and tremelimumab (lgG2 monoclonal antibody formerly known as ticilimumab, CP-675.206) are humanized monoclonal antibodies that bind to human CTLA4 and prevent its interaction with CD80 and CD86. Other negative immune regulators that can be targeted by a similar strategy include programmed cell death 1 (PD1), B and T lymphocyte attenuator, transforming growth factor beta ^Λ , interleukin-10 and vascular endothelial growth factor .
[00200] In some embodiments, the compounds of the present invention can be used in combination with an antiCTLA-4 antibody and an anti-PD-1 antibody. In one embodiment, the combination includes an anti-PD-1 antibody molecule, for example, as described herein, and an anti-CTLA-4 antibody, for example, ipilimumab. Exemplary illustrative doses that can be used include a dose of an anti-PD-1 antibody molecule of about 1 to 10 mg / kg, for example, 3 mg / kg, and a dose of an antiCTLA-4 antibody, for example , ipilimumab, of about 3 mg / kg.
[00201] The compounds of the present invention can be used in combination with antagonists of the PD-1 pathway. In some embodiments, the combination is used to treat a solid tumor or haematological malignancy. PD-1 is another negative regulator of the adaptive immune response that is expressed on active T cells
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47/80 of. PD-1 binds to B7-H1 and B7-DC, and PD-1 coupling suppresses T-cell activation. Antitumor effects have been demonstrated with the blockade of the PD-1 pathway. The molecules of anti-PD-1 antibodies (for example, Nivolumab (Opdivo®), pembrolizumab (Keytruda®) and pidilizumab) and AMP-224 have been reported in the literature as being examples of blockers of the PD-1 pathway that can find use in the present invention. In some embodiments, the PD-1 pathway antagonist is an anti-PD-1 antibody molecule selected from the group consisting of nivolumab, pembrolizumab or pidilizumab.
[00202] In some embodiments, the PD-1 pathway antagonist is an immunoadhesin (for example, an immunoadhesin comprising an PD-LI or PD-L2 PD-1-binding or extracellular portion fused to a constant region (for example, example, an Fc region of an immunoglobulin sequence). In some embodiments, the PD-1 inhibitor AMP-224 (B7-DCIg; Amplimmune; for example, described in W02010 / 027827 and WO2011 / 066342) is a soluble receptor for fusion of PD-L2 Fc that blocks the interaction between PD-1 and B7-H1.
[00203] In some embodiments, the PD-1 pathway antagonist is an inhibitor of PD-L1 or PD-L2. In some embodiments, the PD-L1 or PD-L2 inhibitor is an anti-PD-L1 antibody or an antiPD-L2 antibody. In some embodiments, the anti-PD-LI inhibitor is chosen from YW243.55.S70, MPDL3280A, MEDI-4736, MSB-0010718C or MDX-1105. In some embodiments, the PD-L1 inhibitor is an anti-PD-L1 antibody MSB0010718C. MSB0010718C (also referred to as A09-246-2; Merck Serono) is a monoclonal antibody that binds to PD-L1.
The compounds of the present invention can be used in combination with antagonists of the TIM-3 pathway. In some embodiments, the combination is used to treat a solid tumor or haematological malignancy. In some modalities, the antagonist of
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48/80 via TIM-3 is an anti-TIM-3 antibody. In some embodiments, anti-TIM-3 antibody molecules are described in US 2015/0218274, published on August 6, 2015, entitled Antibody Molecules to TIM-3 and Uses Thereof.
[00205] The compounds of the present invention can be used in combination with antagonists of the LAG-3 pathway. In some embodiments, the combination is used to treat a solid tumor or haematological malignancy. In some embodiments, the antagonist of the LAG-3 pathway is an anti-LAG-3 antibody. In some embodiments, anti-LAG-3 antibody molecules are described in US 2015/0259420, filed on March 13, 2015, entitled Antibody Molecules to LAG-3 and Uses Thereof.
T Cell Receptor Shareholders [00206] The compounds of the present invention can be used in combination with a T cell receptor agonist, such as a CD28 agonist, a 0X40 agonist, a GITR agonist, a CD137 agonist, a CD27 agonist or an HVEM agonist.
[00207] The compounds of the present invention can be used in combination with a CD27 agonist. Illustrative CD27 agonists include an agonistic anti-CD27 antibody, for example, as described in PCT Publication No. WO 2012/004367.
[00208] The compounds of the present invention can be used in combination with a GITR agonist. In some embodiments, the combination is used to treat a solid tumor or haematological malignancy. Illustrative GITR agonists include, for example, GITR fusion proteins and anti-GITR antibodies (for example, divalent anti-GITR antibodies).
TLR shareholders [00209] The compounds of the present invention can be used in combination with a Toll-like receptor agonist. The term receiver
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49/80
Toll-like (or TLR), as used in this document, refers to a member of the family of Toll-like receptor proteins or a fragment thereof that perceives a microbial product and / or initiates an adaptive immune response. In one embodiment, a TLR activates a dendritic cell (DC). Toll-like receptors (TLRs) are a family of pattern recognition receptors that were initially identified as sensors of the innate immune system that recognize microbial pathogens. TLRs comprise a family of conserved membrane transposing molecules containing a leucine-rich repeat ectodomain, a transmembrane domain and an intracellular TIR (Toll / IL-1 R) domain. TLRs recognize distinct structures in microbes, often referred to as PAMPs (molecular patterns associated with pathogens). Binding of the ligand to TLRs invokes a cascade of intracellular signaling pathways that induce the production of factors involved in inflammation and immunity.
[00210] TLR agonists known in the art and finding use in the present invention include, but are not limited to, the following:
[00211] Pam3Cys, a TLR-1/2 agonist;
[00212] CFA, a TLR-2 agonist;
[00213] MALP2, a TLR-2 agonist;
[00214] Pam2Cys, a TLR-2 agonist;
[00215] FSL-1, a TLR-2 agonist;
[00216] Hib-OMPC, a TLR-2 agonist;
[00217] polyribosinic acid: polyribocytoid (Poly I: C), a TLR-3 agonist;
[00218] polyadenosine-polyuridyl acid (poly AU), a TLR-3 agonist;
[00219] Poly-inosinic-polycytidyl acid stabilized with poly-L
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50/80 lysine and carboxymethylcellulose [00220] (Hiltonol®), a TLR-3 agonist;
[00221] monophosphoryl lipid A (MPL), a TLR-4 agonist;
[00222] LPS, a TLR-4 agonist;
[00223] bacterial flagellin, a TLR-5 agonist;
[00224] sialyl-Tn (STn), a carbohydrate associated with mucin MUC1 in several human cancer cells and an agonist of TLR-4;
[00225] imiquimod, a TLR-7 agonist;
[00226] resiquimod, a TLR-7/8 agonist;
[00227] loxoribine, a TLR-7/8 agonist; and [00228] unmethylated CpG dinucleotide (CpG-ODN), a TLR-9 agonist.
[00229] Because of their adjuvant qualities, TLR agonists are preferably used in combinations with other vaccines, adjuvants and / or immunological modulators, and can be combined in various combinations. Thus, in certain modalities, the mono- or di-FCDN compounds that bind to STING and induce STING-dependent TBK1 activation and an inactivated tumor cell that expresses and secretes one or more cytokines that stimulate induction, recruitment and / or maturation of dendritic cells, as described in this document, can be administered in conjunction with one or more TLR agonists for therapeutic purposes.
Antibody Therapy [00230] The compounds of the invention can be used in combination with therapeutic antibodies. In some embodiments, the therapeutic antibody's mechanism of action is Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC). ADCC is a cell-mediated immune defense mechanism whereby an effector cell of the immune system actively smooths a target cell, whose membrane-surface antigens have been linked by anti
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51/80 specific bodies. It is one of the mechanisms through which antibodies, as part of the humoral immune response, can act to limit and contain infection. Classical ADCC is mediated by natural killer cells (NK); macrophages, neutrophils and eosinophils can also mediate ADCC. ADCC is an important mechanism of action of therapeutic monoclonal antibodies, including trastuzumab and rituximab, against tumors. The compounds of the present invention can act to potentiate ADCC.
[00231] The following are an illustrative list of antibodies that can be used in conjunction with the compounds of the present invention.
[00232] Muromonab-CD3, Infliximab, adalimumab, Omalizumab, Daclizumab, Rituximab, Ibritumomab, Tositumomab, Cetuximab, Trastuzumab, Alemtuzumab, Lym-1 Ipilimumab, Vitaxin, Bevacizumab and Abevacizumab and Abevixizumab
[00233] Additional therapeutic antibodies that can be used in combination with compounds of the present invention include a prolactin receptor inhibitor (PRLR), an HER3 inhibitor, an EGFR2 inhibitor and / or EGFR4, an M-CSF inhibitor, an anti-APRIL antibody or an anti-SIRP ^A antibody or an anti-CD47 antibody.
Chemotherapeutic Agents [00234] In additional embodiments of the methods described in this document, the compounds of the present invention are used in combination with chemotherapeutic agents (for example, small molecule pharmaceutical compounds). Thus, the methods further involve administering to the patient an effective amount of one or more chemotherapeutic agents as an additional treatment or combination treatment. In certain embodiments, one or more chemotherapeutic agents are selected from the group consisting of abiraterone acetate, altretamine, anhydrovimblastine, auristatin, bexarotene,
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52/80 bicalutamide, BMS 184476, 2,3,4,5,6-pentafluor-N- (3-fluor-4methoxyphenyl) benzene sulfonamide, bleomycin, N, N-dimethyl-L-valyl-Lvalyl-N-methyl- L-valyl-L-proliI-1-Lproline-tbutilamide, caquectin, cemadotine, chlorambucil, cyclophosphamide, 3 ', 4'-dideshydro-4'-deoxy-8'-norvincaleucoblastine, docetaxol, doxetaxel, cyclophosphamide, carboplatin, carboplatin, cisplatin, cryptoficin, cyclophosphamide, cytarabine, dacarbazine (DTIC), dactinomycin, daunorubicin, decitabine, dolastatin, doxorubicin (adriamycin), etoposide, 5-fluorouracil, finasteride, flutamide, hydroxyurea, hydroxyurea, hydroxyamine, hydroxyamine , enzalutamide, mecloretamine (nitrogen mustard), melphalan, mivobulin isethionate, rhizoxin, sertenef, streptozocin, mitomycin, methotrexate, taxanes, nilutamide, onapristone, paclitaxel, prednimustine, procarbazine, taxpramine, trefoxin, trefoxin, trefoxin, estPR , vinblastine, vincristine, vind sulfate esin and vinflunine.
[00235] In additional embodiments of the methods described in this document, the compounds of the present invention are used in combination with chemotherapeutic agents and / or additional agents to treat the indications as described in the methods contained herein. In some embodiments, the compounds of the present invention are used in combination with one or more agents selected from the group consisting of sotrastaurine, nilotinib, 5- (2,4-dihydroxy-5isopropylphenyl) -N-ethyl-4- (4- ( morpholinomethyl) phenyl) isoxazol-3-arboxamide, dactolisib, 8- (6-Methoxy-pyridi n-3-i) -3-methyl I-1 - (4-piperazin-1-yl-3trifluormethyl-phenyl) - 1,3-dihydro-imidazo [4,5-c] quinolin-2-one, 3- (2,6dichloro-3,5-dimethoxyphenyl) -1 - (6 - ((4- (4-ethylpiperazi n -1 iI) pheniI) amino) pyrimidin-4-iI) -1-methylurea, buparlisib, 8- (2,6-difluoro-3,5dimethoxyphenyl) -N- (4 - (((dimethylamino) methyl)) - 1 H -imidazol-2-yl) quinoxaline5carboxamide, (S) -N1- (4-methyl-5- (2- (1,1,1-trifluor-2-methylpropan-2yl) pi ridi n-4-yl) thiazole- 2-il) pi rrolidi na-1,2-dicarboxamide, (S) -1 - (4Petition 870190083676, of 8/27/2019, page 67/103
53/80 chlorophenyl) -7-isopropoxy-6-methoxy-2- (4- (methyl - ((((1 r, 4S) -4- (4-methyl-3oxopiperazin-1-yl) cyclohexyl) methyl) amino) phenyl) -1,2-dihydroisoquinolin3 (4H) -one, deferasirox, letrozole, (4S, 5R) -3- (2'-amino-2-morpholino-4 '(trifluormethyl) - [4,5 '-bipyrimidin] -6-yl) -4- (hydroxymethyl) -5-methyloxazolidin-2one, (S) -5- (5-chloro-1-methyl-2-oxo-1,2-dihydropyridin-3 -iI) -6- (4-chlorophenyl) 2- (2,4-dimethoxypyrimidin-5-yl) -1-isopropyl-5,6-dihydropyrrolo [3,4-
d] imidazole-4 (1 H) -one, 4 - ((2 - (((1 R, 2R) -2-hydroxycyclohexyl) amino) benzo [d] thiazol-6-yl) oxy) -N- methylpicolin-amide, imatinib mesylate, 2-fluorine-N-methyl-4- (7- (quinolin-6-ylmethyl) imidazo [1,2b] [1,2,4] triazin-2-yl) benzamide, ruxolitinib , panobinostat, osilodrostat, (S) -N - ((S) -1 -cyclohexyl-2 - ((S) -2- (4- (4-fluorbenzoyl) thiazol-2-yl) pyrrolidin-1 yl) -2-oxoethyl) -2- (methylamino) propanamide, (S) -N - ((S) -1-cyclohexyl-2 - ((S) 2- (4- (4-fluorobenzoyl) thiazol-2- il) pyrrolidin-1-yl) -2-oxoethyl) -2 (methylamino) propanamide, sonidegib phosphate, ceritinib, 7-cyclopentylN, N-dimethyl-2 - (((5- (piperazin-1-yl) pyridin-2 -yl) amino) -7H-pyrrolo [2,3-
d] pyrimidine-6-carboxamide, N- (4 - ((1 R, 3S, 5S) -3-amino-5-methylcyclohexyl) pyridin-3-yl) -6- (2,6-difluorphenyl) -5- fluorpicolinamide, 2- (2 ', 3-dimethyl [2,4'-bipyridin] -5-yl) -N- (5- (pyrazin-2-yl) pyridin-2-yl) acetamide, encouragefenib, 7-cyclopentyl -N, N-dimethyl-2 - ((5 - ((1 R, 6S) -9-methyl-4-oxo-3,9 diazabicyclo [4.2.1] -nonan-3-yl) pyridin-2-yl) amino) -7H-pyrrole [2,3-
d] pyrimidine-6-carboxamide, binimetinib, midostaurine, everolimus, 1methyl-5 - ((2- (5- (trifluormethyl) -1 H-imidazol-2-yl) pyridin-4-yl) oxide) -N- ( 4 (trifluormethyl) phenyl) -1 H-benzo [d] imidazole 2-amine, pasireotide diaspartate, dovitinib, (R, E) -N- (7-chloro-1- (1- (4- (dimethylamino) but -2enoyl) azepan-3-yl) -1 H-benzo [d] imidazol-2-yl) -2-methylisonicotinamide, N6 (2-isopropoxy-5-methyl-4- (1-methylpiperidin-4-yl) phenyl ) -N4- (2isopropylsulfonyl) -phenyl) -1 H-pyrazolo [3,4-d] pyrimidine-4,6-diamine, 3- (4- (4 - ((5-chloro-4- ((5-methyl-1 H-pyrazol-3-yl) amino) pyrimidin2-yl) amino) -5-fluor-2-methylphenyl) piperidin-1-yl) tiethane, 5-chloro-N2- (2-fluor5 -methyl-4- (1- (tetrahydro-2H-pyran-4-yl) piperidin-4-yl) phenyl) -N4- (5-methyl-
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H-pi razol-3-i I) pyridine-2,4-diamine, 5-chloro-N2- (4- (1-ethyl peridi n-4yl) -2-fluorine-5- methylphenyl) -N4- (5-methyl-1H-pyrazol-3-yl) pyrimidine-2,4-diamine, valspodar and vatalanib succinate.
[00236] In other embodiments, the compounds of the present invention can be used in combination with a PKC inhibitor, a BCR-ABL inhibitor, an HSP90 inhibitor, a PI3K and / or mTOR inhibitor, a FGFR inhibitor, a PI3K inhibitor, FGFR inhibitor, PI3K inhibitor, cytochrome P450 inhibitor (for example, CYP17 inhibitor), HDM2 inhibitor, aromatase inhibitor, p53 inhibitor and / or one of the interaction of p53 / Mdm2 or a CSF-1R tyrosine kinase inhibitor.
[00237] Suitable preparations include, for example, tablets, capsules, suppositories, solutions - particularly solutions for injection (s.c., i.v., i.m.) and infusion - elixirs, emulsions or dispersible powders. The content of the pharmaceutically active compound (s) should be in the range of 0.1 to 90% by weight, preferably 0.5 to 50% by weight of the composition as a whole, that is, in amounts that sufficient to achieve the dosage range specified below. The specified doses can, if necessary, be administered several times a day.
[00238] The dosage for the combination pairs mentioned above is normally 1/5 of the lowest dose normally recommended up to 1/1 of the dose normally recommended.
[00239] In yet another aspect, the present invention relates to a way to treat a disease or condition associated with, or modulated by, STING in a patient, which includes the step of administering to the patient, preferably a human being. in need of such treatment, a therapeutically effective amount of a compound of the present invention in combination with a therapeutically effective amount of one or more additional therapeutic agents described
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55/80 previously.
[00240] The use of the compound according to the invention in combination with the additional therapeutic agent can occur simultaneously or at staggered times.
[00241] Both the compound according to the invention and the one or more additional therapeutic agents can be present together in one formulation or separately in two identical or different formulations, for example, as a so-called kit of portions.
[00242] Thus, in a further aspect, the present invention provides a combination comprising a compound of formula (I), or one of its salts so long as it is pharmaceutically acceptable, and at least one additional therapeutic agent.
[00243] A further object of the present invention is to provide a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent and one or more of the pharmaceutically acceptable excipients.
[00244] In a further aspect, the invention provides a combination comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent for use in therapy.
[00245] In a further aspect, the invention provides a combination comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent for use in the treatment of a disease or condition in which the STING modulation is beneficial.
[00246] In another aspect, the invention provides a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one additional therapeutic agent
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56/80 onal for use in the treatment of inflammation, allergic and autoimmune diseases, infectious diseases and cancer.
[00247] In a further aspect, the invention provides a method of treating a disease or condition, in which STING modulation is beneficial, in a patient, comprising administering a therapeutically effective amount of a combination comprising a compound of formula (I ), or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent.
[00248] In a further aspect, the invention provides a method of treating inflammation, allergic or autoimmune diseases, infectious diseases or cancer, in a patient, comprising administering a therapeutically effective amount of a combination comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one additional therapeutic agent.
[00249] The pharmaceutically effective amount or the actual therapeutic dosage will, of course, depend on factors known to those skilled in the art, such as age and weight of the patient, route of administration and severity of the disease. In any case, the combination will be administered in dosages and in a manner that allows a pharmaceutically effective amount to be administered based on the patient's unique condition.
[00250] In another aspect, this invention relates to a pharmaceutical composition comprising a compound according to the invention and one or more additional therapeutic agents described above and below, optionally together with one or more carriers and / or diluents inert.
[00251] The other features and advantages of the present invention will become evident from the following more detailed Examples, which illustrate, by way of example, the principles of the invention
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57/80 tion.
Examples and Experimental Data
[00252] The following examples are for illustrative purposes only.
invention and are not intended in any way to limit the scope of the present invention.
[00253] The following abbreviations are used in this document
previously and next:
aq.Bz aqueousbenzoyl Zip code (2-cyanoethyl) - (A /, A / -di-isopropyl)] - phosphoramidite GIVES diode arrangement DCM dichloromethane DDTT 3 - ((A /, A / -dimethyl-aminomethylidene) amino) -3H-1,2,4-dithiazol-3-thione dmf A /, A / -dimethylformamidinyl DMOCP 2-chloro-5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorin DMSO dimethyl sulfoxide DMTr 4,4’-dimethoxytrityl ESI electrospray ionization eq. H hour molar equivalent HPLC high performance liquid chromatography LC liquid chromatography LNA blocked nucleic acid m / z mass to load ratio MeOH methanol min minute MS mass spectrometry NH ₄ OH NH3 solution in water NMR nuclear magnetic resonance
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ppm portions per million s singlet Sun Solvent TEA triethyl amine TEAB triethylammonium bicarbonate TEAF triethylammonium format TFA trifluoroacetic acid ÍRet retention time in minutes uv ultraviolet Vis visible
General Technical Notes [00254] The terms room temperature and room temperature are used interchangeably and indicate a temperature of around 20 Ό, for example, 15 to 25 O.
[00255] As a general rule, ¹ H NMR spectra and / or mass spectra were obtained from the compounds according to the invention. Unless otherwise stated, all chromatographic operations were performed at room temperature. During the synthesis of cyclic dinucleotides, solvent evaporation was typically carried out by rotary evaporation, under reduced pressure, with water bath temperatures not exceeding 35Ό. In addition, during the synthesis of cyclic dinucleotides, reactions were typically carried out under nitrogen or argon.
[00256] The X-ray structures of Example 1.1 (improved binding to human STING compared to Example 1.2) and Example 2.1 (improved binding to human STING compared to Example 2.2) in complex with human STING suggest that both phosphorus atoms have the Rp configuration. In an analogous way, Example 3.1 (improved binding to human STING compared to the other diastereoisomers of Exem is thus assumed)
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59/80 pio 3) and Example 4.1 (improved binding to human STING compared to Example 4.2) also show the Rp configuration on both phosphorus atoms.
A) Analytical Methods
LC-MS analytics:
Method Name: System A [00257] HPLC system: VWR / Hitachi: Pump L-2130; VWR / Hitachi: L-2200 Autosampler; VWR / Hitachi: Column Oven L-2300; VWR / Hitachi: L-2450 Diode Array Detector; Agilent: OpenLab [00258] MS system: Bruker Esquire LC 6000 spectrometer [00259] Column: Kromasil 100-5 Cs, 5 μιτι, 50 mm x 3 mm.
[00260] Flow rate: 0.4 mL / min, 35Ό, UV detection range: 220 300 nm [00261] Mass spectrum: Recorded on a mass spectrometer using negative and positive ESI
Solvents: A: acetonitrile
B: water
C: 20 mM NH4HCO3 (pH 5.5) in water
Gradient: Time % of A % of B % of C%0 2 93 520 60 35 523 95 0 524 2 93 530 2 93 5
[00262] Sample preparation: The samples (2 μΙ_- 20 μΙ_) were dissolved in 87.5 μΙ_ acetonitrile and 262.5 μΙ_ water, injection volume 2 μΙ_ - 5 μΙ Method name: Z011_S03 [00263] Description Device: Agilent 1200 with DA Detector
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60/80 and MS [00264] Column: XBridge C18_3.0x30 mm, 2.5 pm [00265] Column manufacturer: Waters [00266] Description:
Gradient / Solvent % of Sun [H2O % of Sun Flow T [ ^Q C] Time [min] 0.1% NH4OH] [Acetonitrile] [mL / min]0.0 97.0 3.0 2.2 60.0 0.2 97.0 3.0 2.2 60.0 1.2 0.0 100.0 2.2 60.0 1.25 0.0 100.0 3.0 60.0 1.4 0.0 100.0 3.0 60.0
Method name: Z017_S04 [00267] Device description: Agilent 1200 with DA and MS detector [00268] Column: StableBond C18_3.0x30 mm, 1.8 pm [00269] Column manufacturer: Agilent [00270] Description:
Gradient / Solvent % of Sun [H2O % of Sun Flow T [ ^Q C] Time [min] 0.1% TFA] [Acetonitrile] [mL / min]0.0 97.0 3.0 2.2 60.0 0.2 97.0 3.0 2.2 60.0 1.2 0.0 100.0 2.2 60.0 1.25 0.0 100.0 3.0 60.0 1.4 0.0 100.0 3.0 60.0
Method Name: Z018_S04 [00271] Device description: Agilent 1200 with DA and MS Detector [00272] Column: Sunfire C18_3.0x30 mm, 2.5 pm [00273] Column manufacturer: Waters [00274] Description:
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Gradient / Solvent % of Sun [H2O % of Sun Flow T [ ^Q C] Time [min] 0.1% TFA] [Acetonitrile] [mL / min]0.0 97.0 3.0 2.2 60.0 0.2 97.0 3.0 2.2 60.0 1.2 0.0 100.0 2.2 60.0 1.25 0.0 100.0 3.0 60.0 1.4 0.0 100.0 3.0 60.0
Method name: Z035_001 [00275] Device description: Agilent 1200 with DA and MS detector [00276] Column: Atlantis T3, 4.6 x 50 mm, 3 pm [00277] Column manufacturer: Waters [00278] Description:
Gradient / Solvent % of Sun [H2O % of Sun Flow T [ ^Q C] Time [min] 0.02 mol / L TEAF] [Acetonitrile] [mL / min]0.0 98.0 2.0 1.0 25.0 3.0 98.0 2.0 1.0 25.0 12.0 80.0 20.0 1.0 25.0 14.0 0.0 100.0 1.0 25.0 16.0 0.0 100.0 1.0 25.0
Analytical HPLC settings:
Method Name: Configuration A (gradient HPLC) [00279] VWR / Hitachi: pump L-2130; VWR / Hitachi: L-2200 Autosampler; VWR / Hitachi: L-2350 Column Oven (set to 30Ό); VWR / Hitachi: UV-Vis detector with variable wave length L-2400; EZChrom software version 3.3.1 SP1.
[00280] YMC * GEL ODS-A 12 nm (10 pm; 250 x 4 mm) channel A = 20 mM TEAF (pH 6.8) in water; channel B = 100% acetonitrile, 20 mM TEAF (pH 6.8). Gradient: 0 min 100% A; 30 min 100% B; 40 min 100% B, 30 C ^Q; flow rate: 1.0 mL / min; UV 262 nm;
NMR spectroscopy:
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62/80 [00281] Nuclear magnetic resonance (NMR) spectra: For the ¹ H spectra, chemical shifts were used for reference to the DMSO solvent signal (2.50 ppm) or, for D2O measurements, to 0 DSS (4,4-dimethyl-4-silapentane-1sulfonic acid). The ³¹ P NMR spectra were indirectly used for reference by comparing the absolute frequencies of ¹ H / ³¹ P (Bruker BioSpin GmbH, Software: TopSpin, program au: xsi). All ³¹ P NMR spectra were recorded with proton decoupling.
B) Synthesis of Intermediaries
INTERMEDIATE 1.1
5'-OH-2'-F-3'-H-phosphonate-N ⁶ -Bz-2'-deoxyadenosine [00282] A 5'-DMTr-2'-F-3'-CEP-N ⁶ -Bz- 2'-deoxyadenosine (obtained from ChemGenes, 0.654 g, 0.747 mmol) was dissolved in acetonitrile (10 mL) and water (0.033 mL, 1.50 mmol), 2 eq.) At room temperature. Pyridinium trifluoracetate (0.173 g, 0.896 mmol, 1.2 eq.) Was added and the reaction mixture was stirred at room temperature for 10 minutes. Then, tert-butylamine (10 mL,
95.7 mmol) and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was evaporated under reduced pressure, redissolved in anhydrous acetonitrile (25 ml) and evaporated under reduced pressure. The residue was dissolved in dichloromethane (25 ml) and water (0.134 ml, 7.44 mmol, 10 eq.). Dichloroacetic acid (0.555 ml, 6.73 mmol, 9 eq.) In dichloromethane (25 ml) was added and the resulting orange solution was stirred at room temperature for 10 minutes. Pyridine (1.31 mL, 13.6 mmol, 18 eq.) Was added and the reaction mixture was stirred at room temperature for 5 ml
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63/80 nutos.
[00283] The LC-MS analytics of the reaction mixture confirmed the presence of INTERMEDIATE 1.1.
[00284] LC-MS (system A): t _Ret 4.22 min; ESI-MS: 438 [M + H] ⁺ [00285] The solvents were evaporated under reduced pressure and the residue formed an azeotrope with anhydrous acetonitrile (4x15 mL). During the last evaporation procedure, the solution was concentrated to ca. 4 mL of final azeotrope. The anhydrous solution resulting from INTERMEDIATE 1.1 was used immediately in the next sequence of reactions.
INTERMEDIATE 1.2
Linear dimer 5'-OH-LNA-N ⁶ -Bz-adenosine- (3 '->5') - cyanoethylphosphorothioate-2'-F-3'-H-phosphonate-N ⁶ -Bz-2'-deoxyadenosine
[00286] 5'-DMTr-3'-CEP-LNA-N ⁶ -Bz-adenosine (obtained from Exiqon, 0.998 g, 1.27 mmol, 1.7 eq.) Formed an azeotrope with anhydrous acetonitrile (4x15 mL). During the last evaporation procedure, the solution was concentrated to about 4 ml of the final azeotrope. The resulting solution was added to INTERMEDIATE 1.1 (maximum theoretical amount: 0.747 mmol) dissolved in about 4 mL of anhydrous acetonitrile, at room temperature. The reaction mixture was stirred at room temperature for 20 min. A ((A /, A / -dimethylaminomethylidene) amino) -3 / - / - 1,2,4-dithiazoline-3-thione (DDTT) (0.203 g, 0.823 mmol, 1.1 eq.) Was added and the reaction mixture was stirred at room temperature for 30 minutes. The volatiles were evaporated
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64/80 under reduced pressure and the residue was dissolved in dichloromethane (25 ml) and water (0.134 ml, 7.44 mmol, 10 eq.). Dichloroacetic acid (1.11 ml, 13.5 mmol, 18 eq.) In dichloromethane (25 ml) was added and the orange solution was stirred at room temperature for 20 minutes. Pyridine (10 ml) was added and the reaction mixture was stirred at room temperature for 5 minutes.
[00287] The LC-MS analytics of the reaction mixture confirmed the presence of INTERMEDIATE 1.2 as a mixture of diastereoisomers.
[00288] LC-MS (system A): t _Ret 8.32, 8.53 min; ESI-MS: 952 [M + H] ⁺ for each diastereoisomer.
[00289] The bottle was capped, carefully sealed and stored at -700 ° C for 16 hours. The mixture was evaporated under reduced pressure and the residue was coevaporated with anhydrous pyridine (2 x 20 ml) under reduced pressure. Another 40 ml portion of anhydrous pyridine was added and the residue was concentrated under reduced pressure to about 20 ml of total volume. The resulting anhydrous solution of INTERMEDIATE 1.2 was used immediately in the next sequence of reactions.
INTERMEDIATE 1.3
Cyclic dimer LNA-N ⁶ -Bz-adenosine- (3 '->5') - cyanoethyl-phosphorothioate-2'FN ⁶ -Bz-2'-deoxyadenosine- (3 '^ 5') - phosphorothioate
[00290] 2-Chloro-5,5-dimethyl-1,3,2-dioxaphosphorinane 2-oxide (DMOCP) (0.483 g, 2.62 mmol, 3.5 eq.) Was added to INTERME
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DIARY 1.2 crude (maximum theoretical amount: 0.747 mmol) in anhydrous pyridine, in a total volume of about 20 ml_. The resulting mixture was stirred at room temperature for 30 minutes. Water (0.470 ml, 26.1 mmol, 35 eq.) And 3 / 7-1.2-benzodithiol-3-one (0.189 g, 1.12 mmol, 1.5 eq.) Were added and stirring was continued at room temperature. After 30 minutes, the reaction mixture was poured into a solution of sodium hydrogen carbonate (4,500 g, 53.6 mmol) in 150 ml of water and was stirred at room temperature for 5 minutes, followed by the addition of an acetate mixture. ethyl / methyl tert-butyl ether (150 ml, 1: 1). The organic phase was separated and the aqueous phase was further extracted with ethyl acetate / methyl tert-butyl ether (2 x 80 ml, 1: 1). The combined organic phases were dried over anhydrous magnesium sulfate, followed by evaporation of the solvents under reduced pressure and coevaporation with 100 ml of anhydrous toluene. The remaining residue was further dried in vacuo. The crude product was purified by preparative flash chromatography (160 g silica gel, gradient 0-16.7% MeOH in dichloromethane) to produce INTERMEDIATE 1.3 as a crude mixture of diastereoisomers.
[00291] LC-MS (system A): t _Ret = 9.91, 10.08 min; ESI-MS: 966 [M + H] ⁺ for each diastereoisomer.
INTERMEDIATE 2.1
5'-OH-3'-H-phosphonate-LNA-N ⁶ -Bz-adenosine
[00292] INTERMEDIATE 2.1 was prepared in an analogous manner as described for INTERMEDIATE 1.1, starting from 5'-DMTr-3'CEP-LNA-N ⁶ -Bz-adenosine (LNA-A amidite, EQ-0063-1000, obtained from Exiqon). The LC-MS analysis of the crude product confirmed the presence of INTERMEDIARY 2.1.
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66/80 [00293] LC-MS (Z017_S04): t _Ret = 0.62 min; ESI-MS: 448 [M + H] ⁺ .
INTERMEDIATE 2.2
Linear dimer 5'-OH-2'-FN ² -isobutyryl-2'-deoxyguanosine- (3 '^ 5') cyanoethyl-phosphorothioate-3'-H-phosphonate-LNA-N ⁶ -Bz-adenosine
[00294] 5'-DMTr-2'-F-3'-CEP-N ² -isobutyryl-2'-deoxyguanosine (ABCR AB350793, 1.85 g, 2.16 mmol, 1.3 eq.) Formed azeotrope with anhydrous acetonitrile (2x10 ml). The residue was dissolved in anhydrous acetonitrile (10 ml) and concentrated to about 5 ml under reduced pressure. 10 pieces of molecular sieve (3 Á) were added and the resulting mixture was added to INTERMEDIATE 2.1 (maximum theoretical amount: 1.65 mmol) dissolved in about 3 ml_ of anhydrous acetonitrile at room temperature. The reaction mixture was stirred at room temperature for 42 minutes. A ((A /, A / -dimethylaminomethylidene) amino) -3 / 7-1,2,4-dithiazolin-3-thione (DDTT) (381 mg, 1.85 mmol, 1.1 eq.) Was added and the reaction mixture was stirred at room temperature for 30 minutes. The volatiles were evaporated in vacuo and the residue was dissolved in dichloromethane (20 ml) and water (0.299 ml, 16.6 mmol, 10 eq.). Dichloroacetic acid in dichloromethane (6%, 20 ml) was added and the resulting orange solution was stirred at room temperature for 10 minutes. Then, pyridine (15 ml) was added and the reaction mixture was evaporated in vacuo.
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67/80 [00295] The LC-MS analysis of the crude product confirmed the presence of INTERMEDIATE 2.2.
[00296] LC-MS (Z011_S03): t _Ret = 0.60 min; ESI-MS: 934 [M + H] ⁺
INTERMEDIATE 2.3
Cyclic dimer 2'-FN ² -isobutyryl-2'-deoxyguanosine- (3 '^ 5') - cyanoethylphosphorothioate-LNA-N ⁶ -Bz-adenosine- (3 '^ 5') - phosphorothioate
[00297] The crude intermediate 2.2 (maximum theoretical amount: 1.65 mmol) was dissolved in pyridine (36 mL) and the solution concentrated to about 20 mL in vacuo. 2-Chloro-5,5-dimethyl-1,3,2-dioxaphosphorinane 2-oxide (DMOCP) (1134 mg, 6.15 mmol, 3.7 eq.) Was added and the resulting mixture was stirred at room temperature for 15 min Water (1.03 mL, 57.1 mmol, 34 eq.) And 3 / 7-1.2-benzodithiol-3-one (428 mg, 2.54 mmol, 1.5 eq.) Were added and stirring was continued at room temperature. After 5 minutes, the reaction mixture was poured into a sodium hydrogen carbonate solution (4.00 g,
47.6 mmol) in 140 ml of water and was stirred at room temperature for 5 minutes, followed by the addition of a mixture of ethyl acetate / methyl tert-butyl ether (140 ml, 1: 1). The organic phase was separated and the aqueous phase was further extracted with ethyl acetate / methyl tert-butyl ether (1: 1) and several times with dichloromethane. The organic phases were combined, dried over anhydrous magnesium sulfate and the
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68/80 volatiles were removed in vacuo.
[00298] The remaining residue was dissolved in a minimum volume of dichloromethane and purified by preparative flash chromatography (silica gel, DCM / MeOH: gradient 100/0 70/30). The fractions were analyzed by HPLC-MS. The product-containing fractions were combined and the solvent removed in vacuo to produce INTERMEDIATE 2.3 as a crude mixture of diastereoisomers.
[00299] LC-MS (Z017_S04): t _Ret = 0.86 - 0.88 min; ESI-MS: 948 [M + H] ⁺ .
INTERMEDIATE 3.1
Linear dimer 5'-OH-LNA-N ² -dmf-guanosine- (3 '->5') - cyanoethylphosphorothioate-2'-F-3'-H-phosphonate-N ⁶ -Bz-2'-deoxyadenosine
[00300] To a solution of INTERMEDIATE 1.1 (600 mg, 1.17 mmol) was added a solution of Activator 42 (0.1 M in acetonitrile, 24 ml, 2.4 mmol, 2.1 eq.). After several minutes, 5'-DMTr-3'-CEPLNA-N ² -dmf-guanosine- (LNA-G (dmf) -amidite, EQ-0082-1000, obtained from Exiqon, 1.69 g, 1.98 mmol, 1.7 eq.) was added and the mixture was stirred for 45 min at room temperature. A solution of phenylacetyl disulfide (PADS, 710 mg, 2.35 mmol, 2.0 eq.) In pyridine (10 ml) was added and the mixture was stirred for 30 min at room temperature. The volatiles were evaporated in vacuo and the residue formed azeotrope twice with acetonitrile. The residue was dissolved in dichloromethane (14 ml) and water (0.21 ml, 11.7 mmol, 10 eq.).
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Dichloroacetic acid in dichloromethane (6%, 1.14 ml) was added and the resulting orange solution was stirred at room temperature for 20 minutes. Then, a mixture of pyridine (8 ml) and methanol (8 ml) was added and the reaction mixture was evaporated in vacuo. The remaining residue was dissolved in a minimum volume of dichloromethane and purified by preparative flash chromatography (silica gel, DCM / MeOH: gradient 95/5 -> 0/100). The fractions were analyzed by HPLC-MS. The product containing fractions were combined and the solvent removed in vacuo to produce the crude INTERMEDIATE 3.1.
[00301] The LC-MS analysis of the raw material confirmed the presence of INTERMEDIARY 3.1.
[00302] LC-MS (Z017_S04): t _Ret = 0.71 min; ESI-MS: 919 [M + H] ⁺ .
INTERMEDIATE 3.2
Cyclic dimer LNA-N ² -dmf-guanosine- (3 '->5') - cyanoethyl-phosphorothioate2'-FN ⁶ -Bz-2'-deoxyadenosine- (3 '^ 5') - phosphorothioate
[00303] The crude INTERMEDIATE 3.1 (maximum theoretical amount: 0.109 mmol) formed azeotrope twice with the pyridine and was dissolved in pyridine (4 mL). Pivaloyl chloride (0.050 mL, 0.41 mmol, 3.7 eq.) Was added and the resulting mixture was stirred at room temperature for 45 minutes. Elemental sulfur (12 mg, 0.38 mmol, 3.5 eq.) Was added and stirring was continued at room temperature for 45 min. The volatiles were evaporated in vacuo and the
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70/80 residue formed azeotrope twice with toluene. The remaining residue was triturated with acetonitrile and filtered. The filtrate was concentrated under reduced pressure and dried in vacuo to produce INTERMEDIATE 3.2 as a crude mixture of diastereoisomers.
[00304] The LC-MS analytics of the material confirmed the presence of INTERMEDIARY 3.2.
[00305] LC-MS (Z017_S04): t _Ret = 0.78 min; ESI-MS: 933 [M + H] ⁺ .
INTERMEDIATE 4.1
Linear dimer 5'-OH-2'-F-2'-deoxy-inosine- (3 '^ 5') - cyanoethylphosphorothioate-3'-H-phosphonate-LNA-N ⁶ -Bz-adenosine
[00306] 5'-DMTr-2'-F-3'-CEP-2'-deoxy-inosine (MFCD22374068, Astatech, 2 g, 2.59 mmol, 1.7 eq.) Formed azeotrope with anhydrous acetonitrile (3x15 mL). During the last evaporation procedure, the solution was concentrated to about 5 ml of the final azeotrope. The resulting solution was added to INTERMEDIATE 2.1 (maximum theoretical amount: 1.53 mmol) dissolved in about 5 mL of anhydrous acetonitrile, at room temperature. The reaction mixture was stirred at room temperature for 15 minutes. The additional operations for the preparation of INTERMEDIARY 4.1 were performed in an analogous manner as described for INTERMEDIATE 1.2.
[00307] The LC-MS analysis of the raw material confirmed the presence of INTERMEDIATE 4.1.
[00308] LC-MS (system A): t _Ret 7.67 min; ESI-MS: 849 [M + H]
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71/80 [00309] The bottle was capped, carefully sealed and stored at - 70Ό for 16 hours. The mixture was evaporated under reduced pressure and the residue was coevaporated with anhydrous pyridine (2 x 20 ml) under reduced pressure. An additional 40 ml portion of anhydrous pyridine was added and the residue was concentrated under reduced pressure to about 20 ml total volume. The anhydrous solution resulting from INTERMEDIATE 4.1 was immediately used in the next sequence of reactions.
INTERMEDIATE 4.2
Cyclic dimer 2'-F-2'-deoxy-inosine- (3 '->5') - cyanoethyl-phosphorothioateLNA-N ⁶ -Bz-adenosine- (3 '^ 5') - phosphorothioate
[00310] The synthesis of INTERMEDIATE 4.2 was performed in an analogous manner as described for INTERMEDIATE 1.3. The crude reaction mixture was poured into a solution of sodium hydrogen carbonate (6.7 g, 79.8 mmol) in 200 ml of water and was stirred at room temperature for 5 minutes. During this period, a product-containing precipitate was formed and was filtered off. Ethyl acetate (200 ml) was added to the filtrate and the organic phase was separated. The aqueous phase was further extracted three times with ethyl acetate (3 x 200 ml). The combined organic phases were dried over anhydrous magnesium sulfate, followed by evaporation of the solvents under reduced pressure. The precipitate containing the starting product was added, followed by a final coevaporation with 200 ml of anhydrous toluene. After the completion of these operations, the raw material was further dried
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72/80 in vacuo. The crude material was purified by preparative flash chromatography (160 g silica gel, gradient 0-16.7% MeOH in DCM) to produce crude INTERMEDIATE 4.2 as a mixture of diastereoisomers.
[00311] LC-MS (system A): ÍRet = 9.14 min, 9.28 min; ESI-MS: 863 [M + H] ⁺ for each diastereoisomer.
C) Synthesis of Compounds According to the Invention
EXAMPLE 1.1 and EXAMPLE 1.2 (cyclic LNA-adenosine- (3 '^ 5') - phosphorothioate-2'-F-2'-deoxyadenosine- (3 '^ 5') phosphorothioate)
[00312] Methylamine 33% in absolute ethanol (150 mL) was added to the crude cyclic dimer LNA-N ⁶ -Bz-adenosine- (3 '^ 5') - cyanoethylphosphorothioate-2'-FN ⁶ -Bz-2 ' -deoxyadenosine- (3 '^ 5') - phosphorothioate (INTERMEDIATE 1.3; 0.36 g) and the resulting solution was stirred at room temperature for 4 hours. All volatiles were evaporated under reduced pressure and the remaining residue was dried in vacuo. 125 ml of water were added and the resulting suspension was placed in an ultrasonic bath at room temperature. After 15 minutes, this suspension was poured into 100 ml of chloroform and the organic phase was separated. This extraction was repeated two more times with chloroform (2 x 100 mL). The combined organic phases were extracted with 100 ml of water and the combined aqueous phase containing the product was filtered with a 0.45 μιτι Rotilabo®-CME syringe filter (external diameter: 33 mm). The product solution was diluted with water
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73/80 to 245 ml_ and applied to an anion exchange column Q Sepharose® Fast Flow (40 - 165 pm; 125 x 35 mm; - 120 ml_), Cl form, previously regenerated with 2 M sodium chloride and washed with water. The column was washed with water (2 column volumes), followed by a 0 - 1 M gradient of triethylammonium bicarbonate buffer (TEAB, pH 7) in water over 16.7 column volumes, followed by a step gradient with 2 M TEAB, pH 7, over 8.3 column volumes (detection wavelength 254 nm). EXAMPLE 1.1 and EXAMPLE 1.2 eluted with -0.6 M TEAB - -2.0 M TEAB. The product-containing fractions were carefully concentrated under reduced pressure.
[00313] The separation of EXAMPLE 1.1 (second eluent) and EXAMPLE 1.2 (first eluent) was performed by repeated purifications by semi-prepared reverse phase HPLC. The product solution was applied to a YMC * GEL ODS-A 12 nm (10 μιτι; 250 x 16 mm; -50 mL) column, previously equilibrated with 10% acetonitrile, 20 mM triethylammonium format (TEAF, pH 6, 8) in water. Elution was performed with a step gradient of 10%, 12% and 16% acetonitrile, 20 mM TEAF (pH 6.8) in water.
Preparation of EXAMPLE 1.1, sodium salt (according to eluent diastereoisomer) [00314] The desalination of EXAMPLE 1.1, TEA salt, was performed by preparative reversed phase medium pressure liquid chromatography (MPLC). The product solution (-50 mL) was applied to a Merck LiChroprep®RP-18 column (15-25 μιτι; 450 x 25 mm; -220 mL), previously equilibrated with water. The column was washed with water to remove excess TEAF buffer. Then, 2% 2-propanol in water was used to elute the desalted EXAMPLE 1.1. The product-containing fractions were partially concentrated under reduced pressure and subsequently applied to a cation exchange column
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74/80 nica SP Sepharose® Fast Flow (45 - 165 pm; 125 x 35 mm; -120 mL), Na ^{+ form} , previously regenerated with 2 M sodium chloride and washed with water. The column was washed with water until no UV absorbance was detected (detection wavelength 254 nm). The product containing fractions were carefully evaporated under reduced pressure and further dried in vacuo to produce EXAMPLE 1.1 as sodium salt.
[00315] HPLC (configuration A): ÍRet = 10.51 min; ESI-MS: 705 [M + H] ⁺ [00316] ³¹ P NMR (162 MHz, D ₂ O, 303 K): δ 54.5 (s, 1P), 55.0 (s,
1P) ppm.
Preparation of EXAMPLE 1.2, sodium salt (first eluting diastereoisomer) [00317] Desalination and change from triethylammonium to sodium from EXAMPLE 1.2, TEA salt, were carried out in a similar manner as described for EXAMPLE 1.1, TEA salt.
[00318] HPLC (configuration A): ÍRet = 9.25 min; ESI-MS: 705 [M + H] ⁺ [00319] ³¹ P NMR (162 MHz, D ₂ O, 303 K): δ 54.8 (s, 1P), 55.7 (s,
1P) ppm.
EXAMPLE 2.1 and EXAMPLE 2.2 (LNA-adenosine- (3 '^ 5') - phosphorothioate-2'-F-2'-deoxyguanosine- (3 '^ 5') phosphorothioate) cyclic, sodium salt
nh ₂ [00320] TO INTERMEDIATE 2.3 (1300 mg, bad theoretical quantityPetition 870190083676, from 08/27/2019, page 89/103
75/80 max: 1.37 mmol) a 33% solution of methylamine in ethanol (50 ml) was added and the mixture was stirred for 2 h at room temperature. The volatiles were removed in vacuo. The residue was triturated with acetonitrile, filtered, washed with acetonitrile and dried in vacuo. The residue was purified by prep. (Atlantis C18; 20 mM aq. NhLOAc / acetonitrile = 98/2 70/30). The product containing fractions were combined and lyophilized. Two diastereoisomers were obtained in this way which were further purified by prep HPLC. (Bonus; 20 mM aq. Triethylammonium format) / acetonitrile = 98/2 70/30). The fractions containing pure product were combined and lyophilized twice. The triethylammonium salts of the two diastereoisomers were converted into the respective sodium salts, respectively, by dissolving in 1 mL of water, passing through a Bio-Rad Spin column (filled with 250 mg of BT AG 50W-2 resin, hydrogen form 100-200 mesh, conditioned with 3 mL of 1M aq NaOH and then washed neutral with water), eluting with water and lyophilization of the fractions containing product.
[00321] EXAMPLE 2.1 (second eluent diastereoisomer): [00322] LC-MS (Z035_001): t _Ret = 10.73 min; ESI-MS: 721 [M + H] ⁺ [00323] LC-MS (Z018_S04): tRet = 0.60 min; ESI-MS: 721 [M + H] ⁺ [00324] ³¹ P NMR (162 MHz, D2O, 303 K): δ 54.7 (s, 2P) ppm.
[00325] EXAMPLE 2.2 (first eluent diastereoisomer): [00326] LC-MS (Z035_001): t _Ret = 9.09 min; ESI-MS: 721 [M + H] ⁺ [00327] LC-MS (Z018_S04): t _Ret = 0.39 min; ESI-MS: 721 [M + H] ⁺ [00328] ³¹ P NMR (162 MHz, D ₂ O, 303 K): δ 54.7 (s, 1P); 55.0 (s,
1P) ppm.
EXAMPLE 3.1, EXAMPLE 3.2, EXAMPLE 3.3 and EXAMPLE 3.4 (LNA-guanosine- (3 '^ 5') - phosphorothioate-2'-F-2'-deoxyadenosine- (3 '^ 5') phosphorothioate) cyclic, sodium salt
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[00329] To INTERMEDIATE 3.2 (330 mg, maximum theoretical amount: 0.35 mmol), a 33% solution of methylamine in ethanol (16 mL) was added and the mixture was stirred for 1 h at room temperature. The volatiles were removed in vacuo. The residue was triturated with acetonitrile, filtered, washed with acetonitrile and dried in vacuo. The residue was purified by prep. (Atlantis C18; 20 mM aq. NH4OAC / acetonitrile, gradient 98/2 80/20). The fractions containing pure product were combined and lyophilized. Four diastereoisomers could be isolated in this way. The triethylammonium salts of the four diastereoisomers were converted into the respective sodium salts, respectively, dissolving in 1 mL of water, passing through a Bio-Rad Spin column (filled with 250 mg of BT AG 50W-2 mesh hydrogen form) 100-200, conditioned with 1M aq NaOH and then washed neutral with water), eluting with water and lyophilization of the fractions containing product.
[00330] EXAMPLE 3.1 (fourth eluting diastereoisomer):
[00331] LC-MS (Z018S04): t _Ret = 0.60 min; ESI-MS: 721 [M + H] ⁺ [00332] LC-MS (Z035_001): tRet = 11.05 min; ESI-MS: 721 [M + H] ⁺ [00333] ³¹ P NMR (162 MHz, D2O, 303 K): δ 54.4 (s, 1P), 54.7 (s,
1P) ppm.
[00334] EXAMPLE 3.2 (third eluting diastereoisomer): [00335] LC-MS (Z018S04): t _Ret = 0.42 min; ESI-MS: 721 [M + H] ⁺
Petition 870190083676, of 8/27/2019, p. 91/103
77/80 [00336] LC-MS (Z035_001): t _Ret = 9.62 min; ESI-MS: 721 [M + H] ⁺ [00337] ³¹ P NMR (162 MHz, D ₂ O, 303 K): δ 54.4 (s, 2P) ppm.
[00338] EXAMPLE 3.3 (according to eluent diastereoisomer): [00339] LC-MS (Z018S04): t _Ret = 0.26 min; ESI-MS: 721 [M + H] ⁺ [00340] LC-MS (Z035_001): tRet = 8.76 min; ESI-MS: 721 [M + H] ⁺ [00341] ³¹ P NMR (162 MHz, D2O, 303 K): δ 54.8 (s, 1P), 55.9 (s,
1P) ppm.
[00342] EXAMPLE 3.4 (first eluent diastereoisomer): [00343] LC-MS (Z018S04): t _Ret = 0.16 min; ESI-MS: 721 [M + H] ⁺ [00344] LC-MS (Z035_001): tRet = 8.06 min; ESI-MS: 721 [M + H] ⁺ [00345] ³¹ P NMR (162 MHz, D2O, 303 K): δ 54.8 (s, 1P), 56.0 (s,
1P) ppm.
EXAMPLE 4.1 and EXAMPLE 4.2 (LNA-adenosine- (3 '^ 5') - phosphorothioate-2'-F-2'-deoxy-inosine- (3 '^ 5') phosphorothioate) cyclic, sodium salt
[00346] 250 mL of 33% methylamine in absolute ethanol was added to crude INTERMEDIATE 4.2 (0.43 g) and the resulting solution was stirred at room temperature for 5 hours. All volatiles were evaporated under reduced pressure and the residue was further dried in vacuo to produce the crude mixture containing EXAMPLE 4.1 and EXAMPLE 4.2. 250 ml of water were added and the resulting suspension was placed in an ultrasonic bath at am temperature
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78/80 environment. After 15 minutes, this suspension was poured into 125 ml of chloroform and the organic phase was separated. This extraction was repeated two more times with chloroform (2 x 125 ml). The combined organic phases were extracted with 150 ml of water and the aqueous phase containing the combined product was filtered with a 0.45 pm regenerated cellulose (RC) filter (Sartorius Stedim Biotech, outside diameter: 50 mm) to remove particulate components . The product solution was diluted with water to 1100 mL and applied to an anion exchange column Q Sepharose® Fast Flow (40 - 165 pm; 125 x 35 mm; -120 mL), Cl form, previously regenerated with sodium chloride to 2 M and washed with water. The column was washed with water (2 column volumes), followed by a 0 - 1 M gradient of triethylammonium bicarbonate buffer (TEAB, pH 7) in water, over 16.7 column volumes, followed by 1 M of TEAB, pH 7, on 4.2 column volumes (detection wavelength 254 nm). EXAMPLE 4.1 and EXAMPLE 4.2 eluted with -0.4 M TEAB - -0.8 M TEAB. The product-containing fractions were carefully concentrated under reduced pressure.
[00347] The separation of EXAMPLE 4.1 (second eluent) and EXAMPLE 4.2 (first eluent) was performed by preparative reversed phase medium pressure liquid chromatography (MPLC). The product solution (-65 mL) was applied to a Merck LiChroprep®RP-18 column (15-25 pm; 435 x 25 mm; -215 mL), previously equilibrated with 100 mM triethylammonium (TEAF) format in water . Elution was performed with a step gradient of 2%, 3% and 4% 2-propanol, 20 mM TEAF (pH 6.8) in water. The product-containing fractions were carefully concentrated under reduced pressure.
[00348] The other purifications of EXAMPLE 4.1 and EXAMPLE 4.2 were performed by repeated purifications of semi-prepared reverse phase HPLC. The product solutions were applied to a
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79/80 column YMC * GEL ODS-A 12 nm (10 μιτι; 250 x 16 mm; -50 ml_), previously equilibrated with 9 - 12% acetonitrile, 20 mM TEAF (pH 6.8) in water. Elution was performed with a step gradient of 9% and 12% acetonitrile, 20 mM TEAF (pH 6.8) in water (EXAMPLE 4.2) or with 12% acetonitrile, 20 mM TEAF (pH 6.8 ) in water (EXAMPLE 4.1). The product-containing fractions were carefully concentrated under reduced pressure.
Preparation of EXAMPLE 4.1, sodium salt (according to eluent diastereoisomer) [00349] The desalination of EXAMPLE 4.1, TEA salt, was carried out by preparative reversed phase medium pressure liquid chromatography (MPLC). The product solution (-15 mL) was applied to a Merck LiChroprep®RP-18 column (15-25 pm; 450 x 25 mm; -220 mL), previously equilibrated with water. The column was washed with water to remove excess TEAF buffer. Then, 2% 2propanol in water was used to elute EXAMPLE 2.1 desalted. The product-containing fractions were partially concentrated under reduced pressure and subsequently applied to a SP Sepharose® Fast Flow cation exchange column (45 - 165 pm; 125 x 35 mm; -120 mL), Na ^{+ form} , previously regenerated with chloride 2M sodium and washed with water. The column was washed with water until no UV absorbance was detectable (detection wavelength 254 nm). The product containing fractions were carefully evaporated under reduced pressure and further dried in vacuo to produce EXAMPLE 4.1 as sodium salt.
[00350] HPLC (configuration A, UV 250 nm): ÍRet = 10.43 min; ESIMS: 706 [M + H] ⁺ [00351] ³¹ P NMR (162 MHz, D ₂ O, 303 K): δ 54.4 (s, 1P), 54.9 (s,
1P) ppm.
Preparation of EXAMPLE 4.2, sodium salt (first diastereoiPetition 870190083676, of 8/27/2019, page 94/103
80/80 eluting serum) [00352] Desalination and changing the TEA to sodium salt from EXAMPLE 4.2, TEA salt, were carried out in a similar manner as described for EXAMPLE 4.1, TEA salt.
[00353] HPLC (configuration A, UV 250 nm): ÍRet = 9.37 min; ESIMS: 706 [M + H] ⁺ [00354] ³¹ P NMR (162 MHz, D ₂ O, 303 K): δ 54.4 (s, 1P), 54.9 (s,
1P) ppm.

权利要求:
Claims (16)
[1]
1. Compound, characterized by the fact that it presents formula (I)

[2]
2. Compound according to claim 1, characterized by the fact that Base ¹ and Base ² are adenine.
[3]
3. A compound according to claim 1, characterized by the fact that Base ¹ is adenine and Base ² is guanine.
[4]
4. Compound according to claim 1, characterized by the fact that Base ¹ is guanine and Base ² is adenine.
[5]
5. Compound according to claim 1, characterized by the fact that Base ¹ is adenine and Base ² is hypoxanthine.
[6]
6. Stereoisomer (Sp, Sp), (Sp, Rp) or (Rp, Sp) substantially pure, characterized by the fact that it is of a compound, as defined in any one of claims 1 to 5, or a salt thereof.
[7]
Pharmaceutically acceptable salt, characterized in that it is of a compound, as defined in any one of claims 1 to 6.
[8]
8. Pharmaceutical composition, characterized by the fact that it comprises one or more compounds, as defined in any one of claims 1 to 7, or pharmaceutically acceptable salts of the
Petition 870190083676, of 8/27/2019, p. 101/103
2/3 same, optionally together with one or more inert carriers and / or diluents.
[9]
9. Vaccine, characterized by the fact that it comprises a compound, as defined in any one of claims 1 to 7.
[10]
10. Pharmaceutical composition, characterized in that it comprises one or more compounds, as defined in any one of claims 1 to 7, or pharmaceutically acceptable salts thereof, and one or more additional therapeutic agents, optionally together with one or more carriers and / or inert diluents.
[11]
Pharmaceutical composition according to claim 10, characterized in that it comprises a compound, as defined in any one of claims 1 to 7, and one or more additional therapeutic agents.
[12]
12. A compound according to any one of claims 1 to 7, characterized in that it is for use as a medicine.
[13]
13. Use of a compound, as defined in any of claims 1 to 7, characterized by the fact that it is as a vaccine adjuvant.
[14]
14. Use of one or more compounds, as defined in any of claims 1 to 7, characterized by the fact that it is for the preparation of a drug or pharmaceutical composition or vaccine for the treatment of diseases or conditions associated with, or modulated by, , STING, particularly for the treatment of inflammation, allergic or autoimmune diseases, infectious diseases or cancer, in a patient in need of it.
[15]
Compound according to any one of claims 1 to 7, characterized in that it is for use in a method for the treatment of diseases or conditions associated with, or modulates
Petition 870190083676, of 8/27/2019, p. 102/103
3/3 by, STING, particularly for the treatment of inflammation, allergic or autoimmune diseases, infectious diseases or cancer, the method being distinguished by the fact that one or more compounds, as defined in any of claims 1 to 7, are administered to the patient.
[16]
16. Invention, characterized in any form of its embodiments or in any applicable category of claim, for example, product or process or use or any other type of claim encompassed by the matter initially described, revealed or illustrated in the patent application.

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法律状态:
2021-06-15| B15K| Others concerning applications: alteration of classification|Free format text: AS CLASSIFICACOES ANTERIORES ERAM: C07H 21/02 , A61K 31/7084 , A61P 29/00 , A61P 31/00 , A61P 35/00 , A61P 37/00 , A61P 37/08 Ipc: A61K 31/7084 (2006.01), C07H 21/02 (2006.01), A61P |

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2021-10-13| B350| Update of information on the portal [chapter 15.35 patent gazette]|

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申请号 | 申请日 | 专利标题

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EP17162392|2017-03-22|

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EP17162392.9|2017-03-22|

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PCT/EP2018/056656|WO2018172206A1|2017-03-22|2018-03-16|Modified cyclic dinucleotide compounds|

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